PMID- 17075565 TI - Lymphangioleiomyomatosis. AB - BACKGROUND: Lymphangioleiomyomatosis (LAM) is a rare disease that is characterized by proliferation of abnormal smooth muscle-like cells (LAM cells), which leads to the formation of lung cysts, fluid-filled cystic structures in the axial lymphatics, and abdominal tumors. It primarily affects women. METHODS: The authors present a review of large series, registries, and protocols to highlight the prevalence, pathology, clinical features, diagnosis, and treatment options for patients with LAM. RESULTS: LAM commonly presents with progressive breathlessness or with recurrent pneumothorax, chylothorax, or sudden abdominal hemorrhage. Computed tomography (CT) scans show numerous thin-walled cysts throughout the lungs, abdominal angiomyolipomas, and lymphangioleiomyomas. Pulmonary function tests show decreased forced expiratory volume in 1 second (FEV1) and diffusion capacity for carbon monoxide (DLCO). Exercise testing shows gas-exchange abnormalities, ventilatory limitation, and hypoxemia that may occur with near-normal lung function. CONCLUSIONS: No effective treatment currently exists for this progressive disorder. However, recent progress in cancer and smooth muscle cell biology and a better understanding of the factors regulating angiogenesis and lymphangiogenesis may provide a foundation for the development of new therapeutic strategies. PMID- 17075566 TI - Tracheobronchial gland tumors. AB - BACKGROUND: Tracheal tumors are uncommon, making up only 0.2% of all respiratory malignancies in the United States. One consequence of this low incidence is that few centers accumulate meaningful experience. Another is the lack of awareness of effective therapy. Bronchial gland tumors demonstrate oncologic diversity and include benign, low-grade, and high-grade malignant tumors. METHODS: We reviewed the present knowledge of bronchial gland tumors of the trachea, carina, and bronchi, including the epidemiology, presentation, evaluation, tumor types, and treatment options. RESULTS: The malignant bronchial gland tumors, adenoid cystic carcinoma and mucoepidermoid carcinoma, are far more common than benign mucinous cystadenoma or pleomorphic adenoma. Complete resection of localized tumors has excellent long-term results in symptomatic benign tumors. The disease-free survival after resection of malignant tumors is limited by distant metastasis and regional disease, while local recurrence is uncommon. Postoperative mediastinal radiation is now accepted adjuvant therapy. Experience at our institute demonstrates a significant survival advantage for patients with complete resection compared to unresectable patients. CONCLUSIONS: Expanding knowledge of diagnostic evaluation and surgical therapy can improve the long-term survival of patients with tracheobronchial gland tumors. PMID- 17075567 TI - Building community capacity to participate in cancer prevention research. PMID- 17075568 TI - Comparison of long-term outcomes after allogeneic hematopoietic stem cell transplantation from matched sibling and unrelated donors. AB - Long-term survivors of hematopoietic stem cell transplants remain at risk of potentially fatal complications that detract from life quality. Long-term morbidity and mortality were compared between matched recipient cohorts surviving 2 or more years and defined by donor type, HLA matched sibling donor (MSD) or volunteer unrelated donor (URD). Patients were previously entered into the prospective multicenter International Unrelated Search and Transplant Study. Thirty-nine centers provided data on 108 URD and 355 MSD recipients surviving more than 2 years. Long-term survival, performance status, chronic GvHD (c-GvHD), secondary malignancy, endocrine dysfunction, cataracts, bone necrosis and dental pathology were compared between cohorts. Twelve year survival was 77+/-5% for the MSD and 67+/-11% for the URD cohort (P=0.1). Late death occurred in 105 of 463 recipients alive at 2 years, 73 after 355 (21%) MSD and 32 after 108 (30%) URD transplants, P=0.10. Of 105 deaths, the cause was relapse in 60 and unrelated to relapse in 45 cases. Cumulative incidence of extensive c-GvHD (P=0.002), cataracts (P=0.02) and bone necrosis (P=0.02) was higher after URD transplants. No long-term difference in endocrine dysfunction, secondary malignancy and major dental pathology was detected. This landmark study will assist physicians counseling patients pre-transplant and with their long-term care post transplant. PMID- 17075569 TI - Alpha2-adrenoceptor subtypes involved in the regulation of catecholamine release from the adrenal medulla of mice. AB - BACKGROUND AND PURPOSE: This study was carried out to elucidate which alpha(2) adrenoceptor subtypes mediated the inhibition of noradrenaline and adrenaline release from the adrenal medulla of mice. EXPERIMENTAL APPROACH: Isolated adrenal medullae from wild-type and alpha(2A), alpha(2B) and alpha(2C)-adrenoceptor knockout (KO) mice were placed in superfusion chambers. Catecholamine overflow was evoked by 1,1-dimethyl-4-phenylpiperazinium (500 microM) in absence or in presence of the alpha(2)-adrenoceptor agonist medetomidine. The effect of medetomidine was tested in presence of the alpha-adrenoceptor antagonists rauwolscine, WB 4101, spiroxatrine, phentolamine and prazosin. KEY RESULTS: In wild-type mice, medetomidine reduced noradrenaline and adrenaline overflow in a concentration-dependent manner (EC(50) in nM: 1.54 and 1.92; E(max) in % of inhibition: 91 and 94, for noradrenaline and adrenaline, respectively). The pK (D) values of the antagonists for noradrenaline overflow did not correlate with pK(D) values at alpha(2A), alpha(2B), or alpha(2C) binding sites. The pK (D) values of the antagonists for adrenaline overflow correlated positively with pK(D) values at alpha(2C) binding sites (opossum kidney cells). The effect of medetomidine (100 nM) on noradrenaline overflow was significantly reduced in all three alpha(2)KO mice (57, 54, 44 % inhibition, for alpha(2A), alpha(2B), and alpha(2C), respectively), whereas the effect of medetomidine on adrenaline overflow was greatly reduced in alpha(2C)KO mice (14 % inhibition). CONCLUSIONS AND IMPLICATIONS: In the adrenal medulla of mice, all three alpha(2)-adrenoceptor subtypes (alpha(2A), alpha(2B), and alpha(2C)) play an equal role in the inhibition of noradrenaline overflow, whereas the alpha(2C)-adrenoceptor is the predominant alpha(2)-adrenoceptor subtype involved in the inhibitory mechanism controlling adrenaline overflow. PMID- 17075570 TI - Regulatory effect of sulphatides on BKCa channels. AB - BACKGROUND AND PURPOSE: Sulphatides are sulphated glycosphingolipids expressed on the surface of many cell types, particularly neurones. Changes in sulphatide species or content have been associated with epilepsy and Alzheimer's disease. As the large conductance, calcium sensitive K(+) channel (BK(Ca)) are modulated by membrane lipids, the aim of the study was to explore possible effects of sulphatides on BK(Ca) channels. EXPERIMENTAL APPROACH: Using patch-clamp techniques, we studied effects of exogenous sulphatides on BK(Ca) channels expressed in Chinese hamster ovary cells. KEY RESULTS: Sulphatides reversibly increased the whole-cell current and the single channel open probability of BK(Ca) channels dose-dependently. The EC(50) value on the channel at +10 mV was 1.6 microM and the Hill coefficient was 2.5. In inside-out patches, sulphatides increased the single channel open probability from both intra- and extra-cellular faces of the membrane, but more effectively with external application. Furthermore, activation of the channels by sulphatides was independent of intracellular Ca(2+) concentration. Sulphatides also shifted the activation curve of the channels to less positive membrane potentials. Mutant BK(Ca) channels lacking a 59 aminoacid region important for amphipath activation (STREX) were less activated by the sulphatides. CONCLUSIONS AND IMPLICATIONS: Sulphatides are novel activators of BK(Ca) channels, independent of intracellular Ca(2+) or other signalling molecules but partly dependent on the STREX sequence of the channel protein. As changes of sulphatide content are associated with neuronal dysfunction, as in epilepsy and Alzheimer's disease, our results imply that these effects of sulphatides may play important pathophysiological roles in regulation of BK(Ca) channels. PMID- 17075571 TI - In vivo pharmacological resultant analysis reveals noncompetitive interactions between opioid antagonists in the rat tail-withdrawal assay. AB - BACKGROUND AND PURPOSE: Pharmacological resultant analysis is a technique that can detect secondary effects of competitive antagonists in vitro. The utility of pharmacological resultant analysis as a potential tool for the investigation of antagonist interactions in vivo was examined in the present study using two opioid antagonists, naltrexone and CTAP. EXPERIMENTAL APPROACH: Using the experimental design of pharmacological resultant analysis, the well-characterized opioid antagonist naltrexone was examined in the presence of multiple doses of CTAP to block the antinociceptive effects of morphine in the rat warm-water (55(o)C), tail-withdrawal assay. KEY RESULTS: Alone, all doses of naltrexone, CTAP, and CTOP examined blocked the antinociceptive effects of morphine. In the presence of fixed doses of 1 or 10 microg CTAP, increasing doses of naltrexone produced dose-dependent shifts to the right in the morphine dose-response curve. However, a lower dose of naltrexone in combination with 1 or 10 mug CTAP failed to alter the morphine dose-response curve. In the presence of a fixed dose of 0.1 mg kg(-1) naltrexone, CTAP doses produced irregular shifts to the right in the morphine dose-response curves. CONCLUSIONS AND IMPLICATIONS: Resultant analysis was applied and an apparent pK(C) value for CTAP was found to be one log unit higher than the apparent pA(2) value for CTAP, evidence that CTAP may have secondary actions or that a signal transducer function may be altered by the combinations of these antagonists. Taken together, these data suggest pharmacological resultant analysis can reveal novel interactions between antagonists in vivo. PMID- 17075572 TI - Non-genomic effect of testosterone on airway smooth muscle. AB - BACKGROUND AND PURPOSE: Recent studies on blood vessels have provided evidence that testosterone may exert direct effects on smooth muscle. However, an acute effect on airway reactivity has not been shown yet. The aim of this study was to assess the direct effect of testosterone on the responsiveness of male adult rabbit airway smooth muscle (ASM), precontracted with 10 microM acetylcholine, 10microM carbachol or 80 mM KCl. EXPERIMENTAL APPROACH: Contractility studies of rabbit tracheal smooth muscle were performed. KEY RESULTS: Testosterone at concentrations of or above 1 nM had a significant relaxant effect on ASM precontracted with acetylcholine or carbachol, but did not affect ASM precontracted with KCl. The mechanical removal of airway epithelium as well as the inhibition of NO synthetase (by 100microM L-NAME) reduced the relaxation caused by testosterone. The effect of testosterone was not altered by impairing prostanoid synthesis (by 10microM indomethacin). The nitric oxide donor, sodium nitroprusside, had the same relaxant effect on ASM precontracted with either carbachol or KCl. Inhibitors of androgen receptors (10microM flutamide) or DNA transcription (100microM actinomycin D) did not alter the effect of testosterone. Prolonged incubation of ASM with 100 nM or 100 microM testosterone for 24 or 48 h did not alter their responsiveness to acetylcholine. BSA-testosterone (1pM to 100nM) relaxed significantly ASM precontracted with carbachol. The mechanical removal of airway epithelium abolished the relaxant effect of BSA-testosterone. CONCLUSIONS AND IMPLICATIONS: Testosterone relaxes precontracted ASM via an epithelium and NO-mediated way. This effect is mediated via a non-genomic pathway. PMID- 17075574 TI - Constitutive activation of mTOR signaling pathway in post-transplant lymphoproliferative disorders. AB - We examined activation of the mTOR signaling pathway in situ in the primary, normal reactive and patient-derived post-transplant lymphoproliferative disorder (PTLD) tissue samples. We accomplished this analysis by immunohistochemistry on formalin-fixed, paraffin-embedded specimens using a set of highly specific antibodies that permitted us to determine phosphorylation status of the key serines in the mTOR target proteins. Our results demonstrate that the mTOR signaling pathway is activated in reactive tissue in a highly distinct fashion with positive, typically enlarged cells being present primarily in the germinal center and, to a lesser degree, in interfollicular areas with mantle zone being conspicuously negative. We could demonstrate mTOR activation in the lesional cells in the entire spectrum of PTLD subtypes, regardless of their Epstein-Barr virus genome expression status. These data demonstrate the ubiquitous activation of the mTOR signaling pathway in PTLD and indicate that mTOR inhibitors may be effective in treatment and, notably, prevention of PTLDs given their immunosuppressive properties. Furthermore, our results define potential biomarkers of the therapeutic response. Because the constitutive mTOR activation has also been identified in cells isolated from other hematologic malignancies, the ability to examine the in vivo mTOR signaling may have implications reaching beyond the PTLD field. PMID- 17075573 TI - Pro-inflammatory effects of early non-enzymatic glycated proteins in human mesothelial cells vary with cell donor's age. AB - BACKGROUND AND PURPOSE: Diabetes mellitus is prevalent in the elderly population. It is also a disease causing tissue damage through several different mechanisms. Some of these mechanisms are also activated by ageing and this overlap raises questions about how diabetes induces damage in the elderly. Early products of non enzymatic glycation of proteins (Amadori adducts), and the ageing process share the capacity to induce oxidative stress and inflammation in human peritoneal mesothelial cells (HPMCs). We have evaluated the interactions between the age of the donor of the HPMCs and the pro-inflammatory effects of Amadori adducts in those cells. EXPERIMENTAL APPROACH: HPMCs were isolated from 20 individuals (age range 21-81 years) and grown in culture. Using different experimental approaches we determined NF-kappaB dependent transcriptional activity and different NF kappaB-related pro-inflammatory gene and protein expressions in basal (or non stimulated) conditions and after stimulation with two Amadori adducts; highly glycated haemoglobin and glycated bovine serum albumin. KEY RESULTS: Amadori induced effects on NF-kappaB dependent-transcription and on the activity of NOS, COX and several NF-kappaB-related pro-inflammatory genes (iNOS, COX-2, TNF-alpha, IL-1beta, and IL6) diminished as the donor's age increased, being practically absent in cells from donors more than 65 years old. Such decreased effects were inversely correlated with an increased basal expression and activity of these pro inflammatory markers with age. CONCLUSIONS AND IMPLICATIONS: Pro-inflammatory effects of Amadori-adducts in HPMCs were strongly dependent on cell donor's age. This may have significant implications for the mechanisms underlying diabetes induced tissue damage in patients of different ages. PMID- 17075575 TI - Prostaglandin E2 is required for ultraviolet B-induced skin inflammation via EP2 and EP4 receptors. AB - Keratinocytes are the major target of sunlight, and they produce prostaglandin (PG) E(2) upon ultraviolet (UV) exposure. Although indomethacin, one of cyclooxygenase inhibitors, is known to suppress UV-induced acute skin inflammation, it remains uncertain whether endogenous PGE(2) is responsible for UV-induced skin inflammation, and which subtype of PGE(2) receptors mediates this process. UV-induced skin inflammation was investigated by using genetically and pharmacologically PGE(2) receptor-deficient mice. We applied UV-induced skin inflammation model to genetical and pharmacological PGE(2) receptor-deficient mice. We exposed UVB on these mice at 5 kJ/m(2), and examined the ear swelling and the histological findings. We also measured the blood flow using a laser doppler device to assess the intensity of UVB-induced inflammatory change. The UV induced ear swelling at 48 h after exposure was significantly reduced in EP2(-/ ), EP4(-/-) or wild-type mice treated with the EP4 antagonist compared to control mice. Consistently, inflammatory cell infiltration into the local skin, and local blood flow after UV exposure were significantly reduced by EP2 or EP4 signaling blockade. These data suggest that PGE(2)-EP2/EP4 signaling is mandatory in UV induced acute skin inflammation, presumably by enhancing blood flow in the microenvironment. PMID- 17075576 TI - Expression of toll-like receptors and type 1 interferon specific protein MxA in biliary atresia. AB - Viral infection and type I interferon have been implicated in the pathogenesis of biliary atresia (BA), but the expression of toll-like receptors (TLRs) that recognize viruses, as well as of type 1 interferon specific signaling molecules are still unknown in BA. Fresh liver tissues were obtained from patients in early and late stage of BA and from patients with choledochal cyst (CC), as well as from normal controls receiving liver resection for benign lesion other than cholestasis or fibrosis. Archived liver tissues from patients with neonatal hepatitis (NH) were obtained for immunohistochemical studies. TLR2, 3, 4, 7 and 9 that recognized Gram-positive bacteria, double-stranded RNA virus, lipopolysaccharide, single-stranded RNA virus and DNA virus, respectively, were studied. Real-time quantitative reverse transcription polymerase chain reaction (QRT-PCR) was used to quantitate TLR, type I interferon specific molecule MxA, interleukin-6 (IL-6) and IL-8 mRNA expression and immunohistochemistry for TLR 7 and MxA protein staining. These results show that there were significantly higher TLR7 and lower TLR3 and TLR9 mRNA expression in early stage of BA than in CC. MxA mRNA expression was also significantly higher in early stage of BA and in CC than in late stage of BA. Immunoreactive TLR7 and MxA staining was higher in early stage of BA than in late stage of BA, NH and CC, which was associated with significantly higher IL-8 mRNA expression in BA than in CC. The results implicate involvement of TLRs, particularly TLR7, and type 1 specific interferon signaling in the pathogenesis of BA, especially in early stage, which is associated with upregulation of inflammatory cytokines IL-8. PMID- 17075577 TI - Intrahepatic insulin resistance in a murine model of steatohepatitis: effect of PPARgamma agonist pioglitazone. AB - Hepatic insulin resistance is associated with hepatic steatosis and is thought to play an important role in the pathogenesis of steatohepatitis. Using a murine model of steatohepatitis (mice fed a diet deficient in methionine and choline-MCD diet), we tested the effects of the insulin-sensitising, PPARgamma agonist drug pioglitazone (PGZ) on systemic and intrahepatic insulin sensitivity and on liver pathology. Compared to controls, mice fed the MCD diet develop a significant steatohepatitis, have enhanced glucose tolerance and enhanced systemic response to insulin. PGZ did not affect the systemic insulin sensitivity in control or MCD fed mice as assessed in vivo by intraperitoneal glucose or insulin dynamic tests. However, PGZ prevented hepatic fat accumulation and steatohepatitis induced by the MCD diet. This effect was associated with an increased mass of adipose tissue and increased expression and release of adiponectin, while hepatic acyl co-enzyme A oxidase and acyl-co-enzyme A carboxylase, regulating hepatic beta-oxidation of fatty acid, remained unchanged. Steatohepatitis in MCD-diet-fed mice was associated with intrahepatic insulin resistance as shown by a reduced phosphorylation of hepatic insulin receptor, and Akt in response to an insulin stimulus. PGZ to MCD-fed mice restored the activation of the insulin receptor and of the Akt pathway in response to insulin. In conclusion, PGZ alleviates steatosis and steatohepatitis induced by the MCD diet, an effect associated with correction of intrahepatic insulin resistance. PMID- 17075578 TI - The cancer stem cell hypothesis: a work in progress. AB - There is a growing body of evidence that supports the idea that malignant tumors are initiated and maintained by a population of tumor cells that share similar biologic properties to normal adult stem cells. This model, the cancer stem cell (CSC) hypothesis, is based on the observation that tumors, like adult tissues, arise from cells that exhibit the ability to self-renew as well as give rise to differentiated tissue cells. Although the concept of the CSC is not entirely new, advances made over the past two decades in our understanding of normal stem cell biology in conjunction with the recent application of these concepts to experimentally define CSCs have resulted in the identification of CSCs in several human malignancies. PMID- 17075579 TI - Early pathogenic events associated with Sjogren's syndrome (SjS)-like disease of the NOD mouse using microarray analysis. AB - Recently, we reported development of the C57BL/6.NOD-Aec1Aec2 mouse carrying two genetic intervals derived from the NOD mouse. These two genetic regions confer full Sjogren's syndrome (SjS)-like disease in SjS-non-susceptible C57BL/6 mice. The current study was undertaken to apply microarray technology to define the molecular basis underlying onset of SjS-disease in C57BL/6.NOD-Aec1Aec2 mice. Using oligonucleotide microarrays, gene expression profiles of submandibular glands derived from 8- to 12-week-old C57BL/6.NOD-Aec1Aec2 mice and 8-week-old C57BL/6 mice were performed for comparison. Significant differential expressions were determined using the Mann-Whitney U test. Hybridizations using submandibular cDNA probes revealed 75 differentially expressed genes at 8 weeks and 105 differentially expressed genes at 12 weeks of age in C57BL/6.NOD-Aec1Aec2 mice compared to 8-week-old C57BL/6 mice. These genes were related generally to basic cellular activities such as transcription, translation, DNA replication, and protein folding. During the predisease phase, genes upregulated encode proteins associated with the IFN-gamma signal-transduction-pathway (Jak/Stat1), TLR-3 (Irf3 and Traf6) and apoptosis (casp11 and casp3), indicative of chronic proinflammatory stimuli, especially IL-1. Between 8 and 12 weeks of age, sets of clustered genes were upregulated that are associated with adaptive immune responses, especially B cell activation, proliferation and differentiation (Baffr, Taci, Bcma, Blys, April, CD70, CD40L, Traf1, Traf3, Pax5, c-Jun, Elk1 and Nf-kB), and neural receptors (Taj/Troy). Altered gene expressions of TLR3 and TNF superfamily-receptors and ligands during this early phase of SjS suggest a possible viral etiology in the altered glandular homeostasis with an upregulated, possibly overstimulated, B-lymphocyte activation in the early autoimmune response present in the submandibular glands. The importance of NF-kappaB as a critical signal transduction pathway is also suggested but its link is not yet clear. PMID- 17075580 TI - Characterization of pre- and post-treatment pathology after enzyme replacement therapy for Pompe disease. AB - In Pompe disease, a genetic deficiency of lysosomal acid alpha-glucosidase, glycogen accumulates abnormally in the lysosomes of skeletal, cardiac and smooth muscle, and contributes to clinically progressive and debilitating muscle weakness. The present study involved 8 infantile-onset Pompe patients, treated weekly with 10 mg/kg of recombinant human acid alpha-glucosidase (rhGAA). Muscle biopsies were obtained at baseline, 12 and 52 weeks post-treatment to establish an indicator of efficacy. Several histologic strategies were employed to characterize changes in pre- and post-treatment samples, including high resolution light microscopy and digital histomorphometry, electron microscopy, capillary density and fiber type analysis, and confocal microscopy for satellite cell activation analysis. Histomorphometric analysis was performed on muscle samples to assess glycogen depletion in response to enzyme replacement therapy (ERT). The extent of glycogen clearance varied widely among these patient samples, and correlated well with clinical outcome. Low glycogen levels, mild ultrastructural damage, a high proportion of type I fibers, and young age at baseline were all features associated with good histologic response. There was no correlation between capillary density and glycogen clearance, and activated satellite cell levels were shown to be higher in post-treatment biopsies with poor histologic responses. This histopathologic study of infantile Pompe disease provides detailed insight into the cellular progression of the disease and its response to therapy while highlighting a number of methodologies which may be employed to assess regression or progression of the associated pathology. As enzyme replacement therapy becomes more prevalent for the treatment of lysosomal storage diseases, such evaluation of post-treatment pathology will likely become a more common occurrence in the daily practice of pathologists. PMID- 17075581 TI - Hypoxia-inducible factor 1 and VEGF upregulate CXCR4 in glioblastoma: implications for angiogenesis and glioma cell invasion. AB - Hypoxia and hypoxia-inducible factor-1 (HIF-1) play a critical role in glioblastoma multiforme (GBMs). CXCR4 is involved in angiogenesis and is upregulated by HIF-1alpha. CXCR4 is a chemokine receptor for stromal cell-derived factor-1 (SDF-1)alpha, also known as CXCL12. We hypothesized that CXCR4 would be upregulated by hypoxia in GBMs. First, we investigated the expression of HIF 1alpha and CXCR4 in GBMs. CXCR4 was consistently found colocalized with HIF 1alpha expression in pseudopalisading glioma cells around areas of necrosis. In addition, angiogenic tumor vessels were strongly positive for CXCR4. Next, we tested the in vitro effect of hypoxia and vascular endothelial growth factor (VEGF) on the expression of CXCR4 in glioma cell lines and in human brain microvascular endothelial cells (HBMECs). Exposure to hypoxia induced significant expression of CXCR4 and HIF-1alpha in glioma cells, whereas treatment with exogenous VEGF increased CXCR4 expression in HBMECs. We also transfected U87MG glioma cells with an HIF-1alpha construct and observed that CXCR4 was upregulated in these cells even in normoxic conditions. We then used a lentivirus-mediated shRNA expression vector directed against HIF-1alpha. When exposed to hypoxia, infected cells failed to show HIF-1alpha and CXCR4 upregulation. We performed migration assays under normoxic and hypoxic conditions in the presence or absence of AMD3100, a CXCR4 inhibitor. There was a significant increase in the migration of U87MG and LN308 glioma cells in hypoxic conditions, which was inhibited in the presence of AMD3100. These studies demonstrate the critical role played by hypoxia and CXCR4 in glioma cell migration. Based on these studies, we suggest that hypoxia regulates CXCR4 in GBMs at two levels. First, through HIF-1alpha in the pseudopalisading tumor cells themselves and, secondly, by the VEGF-stimulated angiogenic response in HBMECs. We believe this knowledge may lead to a potentially important two-pronged therapy against GBM progression using chemotherapy targeting CXCR4. PMID- 17075582 TI - Computerized morphometry as an aid in determining the grade of dysplasia and progression to adenocarcinoma in Barrett's esophagus. AB - The aims of this study were to use computerized morphometry in order to differentiate between the degree of dysplasia and to predict progression to invasive adenocarcinoma in Barrett's esophagus (BE). Biopsies from 97 patients with BE graded by a consensus forum of expert gastrointestinal pathologists were available for morphometrical analysis. The study group included 36 biopsies negative for dysplasia (ND), none of which progressed to carcinoma; 16 indefinite for dysplasia (IND) and 21 low-grade dysplasia (LGD), of which three progressed in each group and 24 high-grade dysplasia (HGD), of which 15 progressed to invasive carcinoma. Computerized morphometry was used for measuring indices of size, shape, texture, symmetry and architectural distribution of the epithelial nuclei. Low-grade dysplasia was best differentiated from the ND group by nuclear pseudostratification (P=0.036), pleomorphism (P<0.01), and chromatin texture (margination, P<0.01) and from the HGD group by nuclear area (P<0.01), pleomorphism (P<0.01), chromatin texture (margination, P<0.01), symmetry (P<0.01), and orientation (P=0.027). These results were validated on a new set of cases (n=55) using a neural network model, resulting in an accuracy of 89% for differentiating between the ND and LGD groups and 86% for differentiating between the LGD and HGD groups. Within the HGD group, univariate significant predictors of the progression interval to carcinoma were: indices of nuclear texture (heterogeneity: P=0.0019, s.d.-OD: P=0.005) and orientation: P=0.022. Nuclear texture (heterogeneity) was the only independent predictor of progression (P=0.004, hazard=11.54) by Cox's multivariate test. This study proposes that computerized morphometry is a valid tool for determining the grade of dysplasia in BE. Moreover, histomorphometric quantification of nuclear texture is a powerful tool for predicting progression to invasive adenocarcinoma in patients with HGD. PMID- 17075583 TI - Changes in fat-free mass during significant weight loss: a systematic review. AB - OBJECTIVE: To identify the proportion of weight lost as fat-free mass (FFM) by various weight loss interventions. METHODS: Medline and Embase were systematically searched for reliable measurements of FFM before and after weight loss of >10 kg and eligible data were pooled. In a fixed effect model of % FFM loss/weight loss (%FFML), linear regression analysis was used to determine the influence of degree of caloric restriction, exercise, magnitude of weight loss, initial body mass index (BMI) and type of surgery. RESULTS: Data were included from 26 cohorts treated with dietary and behavioral interventions and 29 cohorts of bariatric surgery patients. The degree of caloric restriction was positively associated with %FFML (r (2)=0.31, P=0.006) and in three randomized controlled trials exercise was shown to decrease %FFML. Compared with laparoscopic adjustable gastric banding (LAGB), biliopancreatic diversion (BPD) and roux en Y gastric bypass (RYGB) caused greater log(e) (natural log) %FFML (r (2)=0.453, P<0.001). Differences in log(e) %FFML between surgical procedures were independent of initial BMI and magnitude of weight loss. CONCLUSIONS: The degree of caloric restriction, exercise and rate of weight loss influence the proportion of weight lost as FFM after non-surgical interventions. For surgical interventions, BPD and RYGB result in greater %FFML than LAGB. PMID- 17075585 TI - Efficacy of PDE-5-inhibitors for erectile dysfunction. A comparative meta analysis of fixed-dose regimen randomized controlled trials administering the International Index of Erectile Function in broad-spectrum populations. PMID- 17075587 TI - Influence of demographic factors and biochemical characteristics on the prostate specific antigen (PSA) response to testosterone replacement therapy. PMID- 17075591 TI - Optimizing care for patients with rheumatic disease worldwide: a concept of global rheumatology. PMID- 17075592 TI - Switching tumor necrosis factor inhibitors: an opinion. PMID- 17075598 TI - Strategies to control disease in rheumatoid arthritis with tumor necrosis factor antagonists-an opportunity to improve outcomes. AB - Recent data have shown that disability and joint destruction in rheumatoid arthritis (RA) occur early on in the course of the disease and progress rapidly. It has been shown that in the early stages of RA, disability is attributed to increased disease activity, whereas later in the course of the disease, disability results from irreversible joint damage. These findings support the need to develop treatment strategies that will rapidly bring the disease under control, with the ultimate goal of alleviating symptoms and halting progressive joint damage. A number of such strategies have been evaluated, including the early administration of a biologic agent alone or in combination with high-dose methotrexate. Other, more recent treatment strategies include the tight control of disease activity by targeting specific outcomes necessary for decision making; the use of biologic agents for the treatment of moderate disease; and the induction of remission with a biologic agent early in the course of disease, followed by maintenance therapy using a conventional disease-modifying antirheumatic drug. The substantial positive effect these strategies have on patient outcomes supports the concept that the optimal management of RA involves aggressive early therapy combined with close monitoring of disease progression and modification of ineffective therapeutic strategies. PMID- 17075599 TI - Risk and prevention of tuberculosis and other serious opportunistic infections associated with the inhibition of tumor necrosis factor. AB - Tumor necrosis factor (TNF) is a proinflammatory cytokine that has a key role in the pathogenesis of a variety of autoimmune diseases-including rheumatoid arthritis-and is an important constituent of the human immune response to infection. At present, three anti-TNF agents are approved (in the US and elsewhere) to treat selected autoimmune diseases: infliximab, etanercept, and adalimumab. These biologic agents have been associated with a variety of serious and 'routine' opportunistic infections; however, differences exist in the mechanisms of action of these agents that might confer variation in their associated risks of infection. From a public-health standpoint, the development of active tuberculosis in some patients who receive anti-TNF therapy is a matter of serious concern. Tuberculosis in such patients frequently presents as extrapulmonary or disseminated disease, and clinicians should be vigilant for tuberculosis in any patient taking anti-TNF therapy who develops fever, weight loss, or cough. To prevent the reactivation of latent tuberculosis infection during anti-TNF therapy, clinicians should screen all patients for tuberculosis, and begin treatment if latent infection is found, before anti-TNF therapy is initiated. Specific tuberculosis screening and treatment strategies vary between geographical regions and are reviewed in this document. The screening strategies employed in Europe and North America have reduced the occurrence of anti-TNF associated tuberculosis and are clearly to be recommended, but the role of screening in the prevention of other opportunistic (e.g. fungal) infections is far less certain. PMID- 17075600 TI - Monitoring disease activity of rheumatoid arthritis in clinical practice: contributions from clinical trials. AB - Rheumatoid arthritis is a heterogeneous and progressive autoimmune disease, and patients with this condition show varied responses to treatment. Practical, reliable, individually tailored measures of disease activity and treatment responses are needed. Outcome measures used in randomized, controlled trials, including American College of Rheumatology response criteria and Disease Activity Scores, identify when treatment should be initiated or changed, but can be time consuming and impractical in daily practice. Simplified disease activity indices, abbreviated joint counts and patient-report questionnaires are more-convenient ways to assess therapeutic responses in the clinic. Patient-reported measures of physical function, pain and global disease activity best differentiate the results of active treatment from those of placebo treatment in randomized, controlled trials. Improvements in physical function closely reflect changes in health-related quality of life. Recent trials have demonstrated limited correlations between clinical responses and radiographically demonstrated responses; both should be assessed on a regular basis. It is recommended that three domains be assessed in the clinic for therapeutic responses: patient reported measures of physical function and/or global disease activity; physician assessment of disease activity; and imaging of the hands and/or feet on a biannual basis. Problematic joints and cervical spine involvement should be followed as clinically indicated. Measures of improvement for individually relevant physical activities need to be defined for each patient. PMID- 17075601 TI - Interleukin 6: from bench to bedside. AB - Interleukin (IL)-6 is a pleiotropic cytokine that has important roles in the regulation of the immune response, inflammation, and hematopoiesis. Disruption of IL-6 regulation might, however, affect the immune response and consequently induce immune-mediated inflammatory diseases such as rheumatoid arthritis, systemic juvenile idiopathic arthritis, Castleman disease, and Crohn's disease. Overproduction of IL-6 also contributes, through its roles as a growth factor or an antiapoptotic factor, to the development of malignant diseases such as multiple myeloma and renal cancer. Progress in the study of IL-6 has increased our understanding of the pathological roles of this cytokine in these diseases and provided key evidence that antagonizing its activities can be used as a therapeutic strategy. The application of molecular biology techniques to design monoclonal antibodies as therapeutic agents has made it possible to regulate the IL-6 signal to successfully treat diseases that have so far proved refractory to conventional therapies. Blocking IL-6 actions by use of a humanized antibody, tocilizumab, which targets the IL-6 receptor, has been proven to be therapeutically effective for rheumatoid arthritis, systemic juvenile idiopathic arthritis, Castleman disease and Crohn's disease. In this review, we discuss a paradigm of IL-6 from basic science to clinical use. PMID- 17075602 TI - Diagnosis and treatment of vasculitis of the central nervous system in a patient with systemic lupus erythematosus. AB - BACKGROUND: A 23-year-old white woman with a 3-year history of systemic lupus erythematosus and a 15-month history of lupus nephritis and retinal vasculitis was successfully treated with antibiotics for Pseudomonas aeruginosa pneumonia while on moderate doses of corticosteroids. Even though her pneumonia had improved, she developed acute changes in her mental status that rapidly progressed to encephalopathy with coma. INVESTIGATIONS: Physical examination, fundoscopic examination, laboratory tests for metabolic abnormalities, cerebrospinal fluid analysis, microbiology and serologic testing, electroencephalogram, tests for IgM and IgG anticardiolipin antibodies, neuroimaging including CT of the brain and T1-weighted MRI before and after gadolinium contrast, and flow-attenuated inversion recovery MRI. DIAGNOSIS: Vasculitis of the central nervous system associated with systemic lupus erythematosus. MANAGEMENT: Intravenous methylprednisolone 1,000 mg/day for 3 days, one dose of intravenous pulse cyclophosphamide 750 mg/m(2), intravenous immunoglobulin 400 mg/kg/day for 4 days, plasmapheresis on alternate days for five cycles, and prednisone 40 mg/day. She continued monthly doses of intravenous pulse cyclophosphamide and intravenous pulse methylprednisolone for 6 months, followed by maintenance infusions every 3 months over 2 years. Prednisone was tapered over 18 months. Cyclophosphamide was discontinued after 2 years because of poor bone-marrow tolerance, and was replaced with mycophenolate mofetil 3,000 mg/ day and ciclosporin 50 mg twice daily. PMID- 17075603 TI - Clinical implications of neuroendocrine differentiation in prostate cancer. AB - The cellular signaling pathways of the prostate play a central role in the induction, maintenance, and progression of prostate cancer (CaP). Neuroendocrine (NE) cells demonstrate attributes that suggest they are an integral part of these signaling cascades. We summarize what is known regarding NE cells in CaP focusing on NE cellular transdifferentiation. This significant event in CaP progression appears to be accelerated by androgen deprivation (AD) treatment. We examine biochemical pathways that may impact NE differentiation in a chronological manner focusing on AD therapy (ADT) as a central event in inducing androgen-independent CaP. Our analysis is limited to the common adenocarcinoma pattern of CaP and excludes small-cell and carcinoid prostatic variants. In conclusion, we speculate on the future of treatment and research in this area. PMID- 17075604 TI - Characterization of the androgen receptor in a benign prostate tissue-derived human prostate epithelial cell line: RC-165N/human telomerase reverse transcriptase. AB - The majority of prostate epithelial cell lines stably expressing wild-type (wt) or mutant (mt) androgen receptor (AR) are derived from metastatic prostate cancers. Therefore, the wt AR-expressing RC-165N/human telomerase reverse transcriptase (hTERT) cell line derived from the benign prostate tissue of an African-American patient provides a unique opportunity to assess the functional status of AR in a cellular context not studied before. Although androgen-induced expression of known androgen responsive genes such as PMEPA1, and NDRG1 was observed in RC-165N/hTERT, this cell line expresses prostate-specific antigen (PSA) at significantly lower levels. Chromatin immunoprecipitation assay revealed androgen-dependent binding of AR to androgen response elements of PSA, PMEPA1 and NDRG1 genes. Similarities, as well as differences were noted in the expression of androgen responsive genes between RC-165N/hTERT and LNCaP cells. Comprehensive evaluations of AR functions in RC-165N/hTERT cells suggest that whereas some features of known AR functions are maintained in this benign prostatic tissue derived cell line, other AR functions are not retained. Objective evaluations of similar cell lines will lead to the understanding of AR functions in prostate growth and differentiation. PMID- 17075606 TI - Cortical dysplasia of the left temporal lobe might explain severe expressive language delay in patients with duplication of the Williams-Beuren locus. AB - We report on a new duplication case of 7q11.23, reciprocal of the Williams-Beuren (WB) deletion. The patient, a 13-year-old girl, was ascertained within an array CGH screening of patients with epilepsy and neuronal migration defects. Similarly to the first reported patient, she showed serious difficulties in expressive language in the absence of severe mental retardation and marked dysmorphic features. Magnetic resonance imaging (MRI) of the brain revealed an abnormal development of the cerebral cortex in the left temporal lobe, which showed a simplified gyral pattern, and increased cortical thickness. This finding, which might explain poor language development, suggests that the WB critical region might harbour a dosage-sensitive gene controlling the molecular machinery of neuronal migration, with regional specificity and lateralization. It will be important to confirm our findings in newly diagnosed patients with dup(7)(q11.23). We expect to detect many more patients with the same duplication using widespread clinical implementation of high-resolution genome analysis. PMID- 17075608 TI - Behavioral Modes Arise From a Random Process in the Nudibranch Melibe. AB - Stochastic analysis was applied to observations of spontaneous behavior in the carnivorous mollusc Melibe leonina. Six behaviors were denned that could be easily recognized on inspection and it was found that transitions between each of these behaviors could be fully described by a first-order random process without memory of past behavioral choices. The behaviors are organized by frequency of transition into two modes, a feeding mode and a resting mode. Transitions within modes are more likely than transitions between modes, and the feeding and resting modes are linked by a preferred pair of behavioral transitions. The amount of time spent in the feeding mode is positively correlated with body size, but the average length of a feeding episode is independent of size. This suggests that body size regulates the probability of entry into feeding behavior but does not influence the basic pattern of feeding. In the presence of food the animals express nearly continuous feeding behavior, suggesting that food reduces the probability of exiting the feeding mode. This model of spontaneous behavior in Melibe is used to form hypotheses amenable to further exploration through neurophysiological experiments. PMID- 17075605 TI - Cytosolic proteomic alterations in the nucleus accumbens of cocaine overdose victims. AB - Chronic cocaine use in humans and animal models is known to lead to pronounced alterations in neuronal function in the nucleus accumbens (NAc), a brain region associated with drug reinforcement. Two-dimensional gel electrophoresis was used to compare protein alterations in the NAc between cocaine overdose (COD) victims (n=10) and controls (n=10). Following image normalization, spots with significantly differential image intensities (P<0.05) were identified, excised, trypsin digested and analyzed by matrix-assisted laser desorption ionization-time of flight-time of flight. A total of 1407 spots were found to be present in a minimum of five subjects per group and the intensity of 18 spots was found to be differentially abundant between the groups, leading to positive identification of 15 proteins by peptide mass fingerprinting (PMF). Of an additional 37 protein spots that were constitutively expressed, 32 proteins were positively identified by PMF. Increased proteins in COD included beta-tubulin, liprin-alpha3 and neuronal enolase, whereas decreased proteins included parvalbumin, ATP synthase beta-chain and peroxiredoxin 2. The present data provide a preliminary protein profile of COD, suggesting the involvement of novel proteins and pathways in the expression of this complex disease. Additional studies are warranted to further characterize alterations in the differentially regulated proteins. Understanding the coordinated involvement of multiple proteins in cocaine abuse provides insight into the molecular basis of the disease and offers new targets for pharmacotherapeutic intervention for drug abuse-related disorders. PMID- 17075609 TI - Ethephon Promotes Germination of Echinacea angustifolia and E. pallida in Darkness. AB - Seeds from five lots each of Echinacea angustifolia DC, and E. pallida (Nutt.) Nutt. were germinated in a growth chamber in light (40 MUmol.m(-2). s(-1)) or darkness at 25 degrees C for 16 to 20 d after soaking in 1 mM ethephon or water for 10 min, or moist stratification at 4 - 6 degrees C for two weeks. Either light or ethephon promoted seed germination of E. angustifolia and E. pallida, in comparison with darkness in nine of ten lots. Ethephon in the dark had similar or greater germination percentages than water with light. Ethephon with light improved germination in three of ten lots compared with ethephon in the dark. The effect of cold, moist stratification in comparison with darkness varied by seed lot. Five lots of E. purpurea (L.) Moench were tested; however, no treatment differences were measured. The finding that ethethon promoted E. angustifolia and E. pallida seed germination in darkness could be useful in the cultivation of these two species. Chemical name used: 2-chloroethylphosphonic acid (ethephon). PMID- 17075610 TI - Cognitive Processing Therapy for Acute Stress Disorder Resulting From an Anti-Gay Assault. AB - This case study describes Cognitive Processing Therapy (CPT) with a 30-year-old gay man with symptoms of acute stress disorder (ASD) following a recent homophobic assault. Treatment addressed assault-related posttraumatic stress disorder symptoms and depressive symptoms. Also addressed were low self-esteem, helplessness, and high degrees of internalized homophobia. Client symptomatology was tracked using the PTSD Symptom Scale and the Beck Depression Inventory over the course of 12 sessions and for a 3-month posttermination session. Symptoms were significantly reduced by the end of the 12-week therapy and were maintained at 3-month follow-up. This case highlights the utility of this therapy in targeting both ASD symptoms and internalized homophobia relating to experiencing a hate crime-related assault. The authors elaborate on theoretical and applied issues in adapting a structured cognitive-behavioral intervention to the treatment of ASD symptoms associated with experiencing a hate crime. PMID- 17075611 TI - Youth With Epilepsy: Development of a Model of Children's Attitudes Toward Their Condition. AB - A model of children's attitudes toward their epilepsy was tested in 173 children (9-14 years) with epilepsy and their parents. Predictor variables tested were child characteristics, family mastery, child worry, child self-efficacy for seizure management, child psychosocial care needs, and seizure variables. Data were analyzed using structural equation modeling, leading to a revised model in which less child worry, greater family mastery, and greater child seizure self efficacy were directly related to more child positive attitudes. Discussion focuses on potential targets for psychosocial interventions aimed at improving attitudes toward epilepsy. PMID- 17075613 TI - Clinical features of muscle dysmorphia among males with body dysmorphic disorder. AB - Muscle dysmorphia - a pathological preoccupation with muscularity - appears to be a form of body dysmorphic disorder (BDD) with a focus on muscularity. However, little is known about muscle dysmorphia in men with BDD, and no study has compared men with BDD who do and do not report muscle dysmorphia. To explore this issue, we reviewed the histories of 63 men with BDD; we compared those rated as having a history of muscle dysmorphia with those who had BDD but not muscle dysmorphia in several domains. The 14 men with muscle dysmorphia resembled the 49 comparison men in demographic features, BDD severity, delusionality, and number of non-muscle-related body parts of concern. However, those with muscle dysmorphia were more likely to have attempted suicide, had poorer quality of life, and had a higher frequency of any substance use disorder and anabolic steroid abuse. Thus, muscle dysmorphia was associated with greater psychopathology. PMID- 17075614 TI - Social anxiety in body dysmorphic disorder. AB - Although clinical impressions suggest that patients with body dysmorphic disorder (BDD) experience distress in social situations, social anxiety in BDD has received little investigation. This study examined social anxiety in 81 patients with BDD and change in social anxiety with pharmacotherapy. Subjects completed the Social Avoidance and Distress Scale (SADS) and were assessed with measures of BDD symptomatology. Participants in a placebo-controlled fluoxetine trial completed measures at baseline and endpoint. The mean SADS score was 1.3 SD units higher than nonclinical sample means but consistent with other clinical sample means. Social anxiety was significantly correlated with BDD severity. Greater depressive symptoms as well as comorbid avoidant personality disorder, but not comorbid social phobia, were also associated with higher SADS scores. Social anxiety did not improve more with fluoxetine than placebo, yet it improved significantly more in fluoxetine responders than in nonresponders. Understanding social anxiety in BDD has implications for reducing rates of misdiagnosis and treatment dropout. PMID- 17075616 TI - The development of cryoprobe nuclear magnetic resonance spectroscopy for the rapid detection of organic contaminants in potable water. AB - The detection of trace levels of a range of organic contaminants (including pesticides, toxins and an explosive) in potable water, using cryoprobe NMR spectroscopy with limited sample preparation and rapid acquisition times, is described. Emphasis is placed on the applicability of NMR spectroscopy for use in emergency scenarios as the unbiased nature of the technique facilitates the detection and characterization of unknown compounds at levels as low as 50 microg L(-1). PMID- 17075617 TI - Air contaminants in a submarine equipped with air independent propulsion. AB - The Swedish Navy has operated submarines equipped with air independent propulsion for two decades. This type of submarine can stay submerged for periods far longer than other non-nuclear submarines are capable of. The air quality during longer periods of submersion has so far not been thoroughly investigated. This study presents results for a number of air quality parameters obtained during more than one week of continuous submerged operation. The measured parameters are pressure, temperature, relative humidity, oxygen, carbon dioxide, hydrogen, formaldehyde and other volatile organic compounds, ozone, nitrogen dioxide, particulate matter and microbiological contaminants. The measurements of airborne particles demonstrate that air pollutants typically occur at a low baseline level due to high air exchange rates and efficient air-cleaning devices. However, short-lived peaks with comparatively high concentrations occur, several of the sources for these have been identified. The concentrations of the pollutants measured in this study do not indicate a build-up of hazardous compounds during eight days of submersion. It is reasonable to assume that a substantial build-up of the investigated contaminants is not likely if the submersion period is prolonged several times, which is the case for modern submarines equipped with air independent propulsion. PMID- 17075618 TI - Indoor air pollution by 2-ethyl-1-hexanol in non-domestic buildings in Nagoya, Japan. AB - 2-Ethyl-1-hexanol is a possibly causative chemical in sick building symptoms, although 2-ethyl-1-hexanol has received little attention as a hazardous substance in studies on indoor air pollution. Airborne 2-ethyl-1-hexanol concentrations were measured from 2002 to 2004 in 99 rooms of 42 non-domestic buildings in Nagoya, Japan. The diffusive sampling method is effective for the measurement of a low level of 2-ethyl-1-hexanol in indoor air. The geometric mean (geometric standard deviation) of 2-ethyl-1-hexanol concentrations was 16.5 (5.4) microg m( 3) in indoor air and 1.9 (2.2) microg m(-3) in outdoor air. The maximum concentration of 2-ethyl-1-hexanol in indoor air and outdoor air was 2709 microg m(-3) and 12.4 microg m(-3), respectively. Fewer rooms in a small number of new buildings showed high concentrations of 2-ethyl-1-hexanol, while low concentrations were observed in many rooms of these buildings as well as the other new buildings. The room-to-room concentrations of 2-ethyl-1-hexanol in each building exhibited a wide variation. The geometric mean of the 2-ethyl-1-hexanol concentrations was significantly higher for indoor air than for outdoor air (p < 0.01). The correlation of the 2-ethyl-1-hexanol concentrations between indoor and outdoor air was not significant. Mechanical ventilation was effective in the temporary reduction of indoor 2-ethyl-1-hexanol level. These results suggest that the predominant source of 2-ethyl-1-hexanol was indoor areas. PMID- 17075619 TI - Spatial interpolation of ambient ozone concentrations from sparse monitoring points in Belgium. AB - Due to scientific interest on the one hand and political and regulatory obligations on the other hand the monitoring of ozone in the troposphere is an important issue. To this end, in Belgium as in many other countries, a fixed network of monitoring stations is operated. In order to estimate the ozone concentrations over the whole territory, a model is needed to spatially complement the sparse measurements. This paper describes the development of an interpolation scheme which is aimed at fast operational use. The model uses the population density as auxiliary data to remove a spatial trend due to titration by nitric oxide. The residuals are interpolated by kriging. As a benchmark the inverse distance weighting interpolation method is used with and without the detrending. The proposed model systematically improves the interpolation and makes a significant difference when estimating human exposure to ozone. It is generic in design, easy to implement and flexible to changes in the monitoring network. PMID- 17075620 TI - A two-stage cyclone using microcentrifuge tubes for personal bioaerosol sampling. AB - Personal aerosol samplers are widely used to monitor human exposure to airborne materials. For bioaerosols, interest is growing in analyzing samples using molecular and immunological techniques. This paper presents a personal sampler that uses a two-stage cyclone to collect bioaerosols into disposable 1.5 ml Eppendorf-type microcentrifuge tubes. Samples can be processed in the tubes for polymerase chain reaction (PCR) or immunoassays, and the use of multiple stages fractionates aerosol particles by aerodynamic diameter. The sampler was tested using fluorescent microspheres and aerosolized fungal spores. The sampler had first and second stage cut-off diameters of 2.6 microm and 1.6 microm at 2 l min( 1)(geometric standard deviation, GSD = 1.45 and 1.75), and 1.8 microm and 1 microm at 3.5 l min(-1)(GSD = 1.42 and 1.55). The sampler aspiration efficiency was >or=98% at both flow rates for particles with aerodynamic diameters of 3.1 microm or less. For 6.2 microm particles, the aspiration efficiency was 89% at 2 l min(-1) and 96% at 3.5 l min(-1). At 3.5 l min(-1), the sampler collected 92% of aerosolized Aspergillus versicolor and Penicillium chrysogenum spores inside the two microcentrifuge tubes, with less than 0.4% of the spores collecting on the back-up filter. The design and techniques given here are suitable for personal bioaerosol sampling, and could also be adapted to design larger aerosol samplers for longer-term atmospheric and indoor air quality sampling. PMID- 17075621 TI - Assessment of dermal exposure to benzene and toluene in shoe manufacturing by activated carbon cloth patches. AB - OBJECTIVES: The aim of this investigation was to use activated carbon cloth (ACC) patches to study the probability and extent of dermal exposure to benzene and toluene in a shoe factory. METHODS: Inhalation and dermal exposure loading were measured simultaneously in 70 subjects on multiple days resulting in 113 observations. Dermal exposure loading was assessed by ACC patches attached to likely exposed skin areas (e.g. the palm of the hand and abdomen). A control patch at the chest and an organic vapor monitor (OVM) were used to adjust the hand and abdomen patches for the contribution from the air through passive absorption of benzene and toluene on the ACC patches. Systemic exposure was assessed by quantification of unmetabolized benzene (UBz) and toluene (UTol) in urine. RESULTS: Mean air concentrations for the study population were 1.5 and 7.5 ppm for benzene and toluene, respectively. Iterative regression analyses between the control patch, OVM and the dermal patches showed that only a small proportion of the ACC patches at the hand had likely benzene (n = 4; mean 133 microg cm(-2) h(-1)) or toluene (n = 5; mean 256 microg cm(-2) h(-1)) contamination. Positive patches were exclusively observed among subjects performing the task of gluing. Significant dermal exposure loading to the abdomen was detected only for toluene (n = 2; mean 235 microg cm(-2) h(-1)). No relation was found between having a positive hand or abdomen ACC patch and UBz or UTol levels. In contrast a strong association was found between air levels of benzene (p = 0.0016) and toluene (p < 0.0001) and their respective urinary levels. CONCLUSIONS: ACC patches are shown to be a useful technique for quantifying the probability of dermal exposure to organic solvents and to provide estimates of the potential contribution of the dermal pathway to systemic exposure. Using ACC patches we show that dermal exposure to benzene and toluene in a shoe manufacturing factory is probably rare, and when it occurred exposures were relatively low and did not significantly contribute to systemic exposure. PMID- 17075622 TI - Trace metal bioaccumulation in eight common coastal Australian polychaeta. AB - Whole tissue trace metal concentrations of ten metals in eight common coastal Australian polychaete species collected from uncontaminated locations were measured. The mean concentration range for each trace metal was: Mn: 2.6-13 microg g(-1); Co: 0.8-4.6 microg g(-1); Cu: 3.4-26 microg g(-1); Zn: 47-225 microg g(-1); As: 18-101 microg g(-1); Se: 2.2-20.4 microg g(-1); Ag: 0.03-2.5 microg g(-1); Cd: 0.07-17 microg g(-1); Hg: 0.08-0.88 microg g(-1) and Pb: 0.09 3.2 microg g(-1)dry mass. Principal components analysis of trace metal signatures revealed that the habitat, i.e. exposed coast sand or rock, estuarine sand or estuarine mud substrate in which a polychaete species was found, had a significant influence on the bioaccumulation of six trace metals (Mn, Cu, Zn, Ag, Cd and Pb). However, there is no clear relationship between trace metal concentrations in substrates and polychaetes. The results of the current study contribute to a reference dataset of polychaete species-specific natural background trace metal concentrations for use in determining the extent of trace metal contamination by urban and industrial sources. PMID- 17075623 TI - Variability in concentrations of potentially toxic elements in urban parks from six European cities. AB - Use of a harmonised sampling regime has allowed comparison of concentrations of copper, chromium, nickel, lead and zinc in six urban parks located in different European cities differing markedly in their climate and industrial history. Wide concentrations ranges were found for copper, lead and zinc at most sites, but for chromium and nickel a wide range was only seen in the Italian park, where levels were also considerably greater than in other soils. As might be expected, the soils from older cities with a legacy of heavy manufacturing industry (Glasgow, Torino) were richest in potentially toxic elements (PTEs); soils from Ljubljana, Sevilla and Uppsala had intermediate metal contents, and soils from the most recently established park, in the least industrialised city (Aveiro), displayed lowest concentrations. When principal component analysis was applied to the data, associations were revealed between pH and organic carbon content; and between all five PTEs. When pH and organic carbon content were excluded from the PCA, a distinction became clear between copper, lead and zinc (the "urban" metals) on the one hand, and chromium and nickel on the other. Similar results were obtained for the surface (0-10 cm depth) and sub-surface (10-20 cm depth) samples. Comparisons with target or limit concentrations were limited by the existence of different legislation in different countries and the fact that few guidelines deal specifically with public-access urban soils intended for recreational use. PMID- 17075625 TI - The use of transgenic and knock-out mice in the investigation of ocular surface cell biology. AB - The transgenic and knock-out mice created by transgenesis and gene targeting techniques are very useful for elucidating the pathophysiology of human diseases caused by altered genetic functions. Many of the experimental mouse lines exhibit ocular surface disorders. However, embryonic lethality and congenital defects found in many of the transgenic and knock-out mice preclude their use for studying the consequences of altered genetic functions in adult animals. To circumvent these difficulties, we have established binary inducible mouse models, using the corneal keratocyte-specific keratocan promoter, and the tetracycline inducible gene expression system (reverse tetracycline transcription activator- rtTA). In these models, the animals function normally until they are fed doxycycline, thus inducing the overexpression of inserted transgenes by keratocytes. We have also developed inserted rtTA and Cre reporter gene constructs to create genetically modified mouse lines that have tissue-specific gene alterations to study acquired conditions, e.g., wound healing and irregular hormone and cytokine signaling that offsets homeostasis in adults. Furthermore, the genes that are ubiquitously expressed in many tissues can be specifically ablated solely in ocular surface tissues to examine their function, since the loss of such a gene in ocular surface tissues will not be life-threatening. It is noteworthy that these altered mouse lines can also be used as models for the development of therapeutic treatment regimens of diseases using gene therapy and stem cell strategies. PMID- 17075626 TI - Challenges and pitfalls in clinical trials of treatments for dry eye. AB - Clinical trials to evaluate the effectiveness of therapy for dry eye disease are challenging because of the nature of the disease, the multiple palliative methods used by patients to control their symptoms, and the potential limitations of the techniques available to evaluate therapeutic outcomes. This review identifies some of the pitfalls encountered in recent clinical trials in dry eye disease. The peculiarities of dry eye disease with respect to symptoms, signs, and patho- physiological changes are discussed. Potential problems that apply to all clinical trials, including patient selection, randomization in small populations, and assessment of the placebo effect, are presented with respect to dry eye clinical trials. Considerations regarding study design address inclusion/exclusion criteria and selection of outcome measures. Special attention is given to methods of symptom analysis, techniques of staining of the ocular surface, and grading systems for surface staining. Alternative methods to the standard clinical trial are mentioned to place them in perspective for overall evaluation of dry eye disease therapy. Finally, caveats are provided to encourage investigators to vigorously conduct future clinical trials in dry eye therapy. PMID- 17075629 TI - Personal profile. PMID- 17075627 TI - Anti-inflammatory therapy of dry eye. AB - As tear secretion and tear clearance decrease in the dry eye, an inflammatory response is initiated on the ocular surface that appears to involve both soluble and cellular mediators. Although the traditional approach to treating dry eye is to hydrate and lubricate the ocular surface with artificial tears, symptoms and/or sight-threatening corneal disease may persist in some patients on such aqueous enhancement therapies. In these patients, treatment with anti inflammatory agents, such as cyclosporin A, corticosteroids, tetracyclines, or autologous serum, may be considered. Results of studies investigating the use of these agents are discussed. During treatment, patients should be conscientiously monitored for adverse effects. PMID- 17075630 TI - The Ocular Surface Center at the University of Southern California. PMID- 17075632 TI - If there were no tears, the dot in the center of the eye would not be called a pupil. PMID- 17075633 TI - Growth factor, cytokine and protease interactions during corneal wound healing. AB - Healing of corneal injuries is an exceptionally complex process involving the integrated actions of multiple growth factors, cytokines, and proteases produced by epithelial cells, stromal keratocytes, inflammatory cells, and lacrimal gland cells. Following corneal injury, basal epithelial cells migrate and proliferate in response to chemotactic cytokines and mitogenic growth factors, including epidermal growth factor and keratinocyte growth factor. Simultaneously, keratocytes adjacent to the injured area undergo apoptosis under the Fas/Fas ligand system, while more distant keratocytes transform into activated fibroblasts and migrate into the wound, where they begin synthesizing new extracellular matrix components that form the scar tissue under the dominant influence of the TGFb/ CTGF system. Epithelial cells and activated stromal fibroblasts also secrete growth factors and cytokines that have paracrine and autocrine functions. Corneal repair proceeds for the next several weeks to months, during which time the gene expression profile slowly returns to the pre injury pattern and the provisional scar matrix slowly remodels by actions of matrix metalloproteinases. While minor epithelial injuries heal by regeneration of normal architecture, large stromal injuries heal by repair with irregular scar tissue that impairs the optical properties of the cornea.Also, if the integrated regulation of the wound healing process is interrupted at any point, the wound fails to heal properly and a corneal ulcer develops. Better understanding of the cellular and molecular changes that occur during repair of corneal wounds will provide the opportunity to design agents that selectively modulate key phases of corneal wound healing, resulting in scars that more closely resemble normal corneal architecture. PMID- 17075634 TI - Current treatment options for conjunctival and corneal intraepithelial neoplasia. AB - Intraepithelial neoplasia of the cornea and conjunctiva (CIN) and squamous cell carcinoma (SCC) lie on a continuum of the same dysplastic process. The etiology of this disease is most likely multifactorial, involving such factors as age, fair pigmentation, ultraviolet light exposure, human papillomavirus, and human immunodeficiency virus (HIV). It is known that CIN and SCC have a high recurrence rate after excision alone. Cryotherapy, radiation, and chemotherapeutics have been used after excision to reduce recurrence rates. More recently, mitomycin-C, 5-fluorouracil, and interferon-alpha-2b have been successfully employed alone against CIN and SCC, thereby eliminating the need for surgical excision altogether. The various treatments for CIN and SCC are reviewed and discussed. PMID- 17075635 TI - Keratopathy in congenital aniridia. AB - Although the most apparent clinical finding in aniridia is the absence of iris tissue, additional ocular structures are often affected. Mutations of the Pax 6 gene, which is important for eye development, have been identified in families with members affected by aniridia. Poor vision in aniridic eyes may be the result of macular hypoplasia, nystagmus, amblyopia, cataracts, glaucoma, and corneal disease, termed aniridic keratopathy. Advances in surgical techniques have improved management of some of the visually disabling manifestations of aniridia, but aniridic keratopathy remains a significant source of visual loss. We have conducted a large, retrospective study of patients with aniridia to gain information about the natural course of aniridic keratopathy. In this paper, we report the results of our study, as well as findings reported in the literature. Penetrating keratoplasty alone has not been a successful treatment for severe stromal scarring, as it does not treat the underlying epithelial causes of corneal disease. However, it has been successful in corneas that have achieved stable epithelium following limbal stem cell transplantation. PMID- 17075636 TI - Compliance issues in manufacturing of drugs. AB - The therapeutic process relies on the assumption that the prescription written by the clinician is what is dispensed to the patient. In this article, we review the many factors involved in the manufacture of pharmaceutical products so that they consistently meet quality standards as approved by regulatory agencies. Industry and government agencies have developed procedures and laws to assure quality in Chemistry, Manufacturing and Controls (CMC) for pharmaceuticals, addressing conditions that may be harmful to patient health. This includes procedures for receipt of materials, production, packaging, labeling, quality control, release, storage and distribution, known as current Good Manufacturing Practice. Special control considerations for ophthalmic products include foreign particulate matter control, preservative effectiveness tests, sterility and leaching of chemicals from the packaging into the product. We also describe the "form, fill and seal" manufacture process as used in unit-dose, non-preserved ophthalmic products; product expiration dates; stages of drug development; and financial considerations for pharmaceutical companies. PMID- 17075637 TI - How are tear lubricant products regulated in the US? PMID- 17075638 TI - Personal profile. PMID- 17075639 TI - The Ocular Surface Center at the Baylor College of Medicine, Houston. PMID- 17075641 TI - When were the goblet cells of the conjunctiva first described? PMID- 17075642 TI - Meibomian gland function and the tear lipid layer. AB - The meibomian glands of the lid produce a lipid material whose synthesis is dependent on neuronal, hormonal, and vascular factors. This lipid material is fluid, spreads easily, is a surfactant as well as an aqueous barrier and must remain functional after a blink. To satisfy these requirements, the meibomian lipids have a specific composition. Even after delivery it may be modified by lipases produced by ocular bacteria, and modifications in the lipid components can lead to unique disease states. For example, bacteria may degrade lipids, producing an unstable tear film and irritating free fatty acids; and hormonal imbalances may alter lipid profiles to destabilize the tear film and produce evaporative dry eye. PMID- 17075643 TI - Meibomian gland dysfunction: a clinical scheme for description, diagnosis, classification, and grading. AB - Although meibomian gland disease (MGD) is one of the most common disorders encountered in ophthalmic practice, there has been no descriptive system consistently accepted to clinically characterize the anatomical and correlative biochemical alterations that occur in this condition. The purpose of this review is to synthesize a clinical description of meibomian gland disease and to provide a scheme for diagnosis, classification, and quantification that will be of value in the clinical setting and in the conduct of clinical trials. PMID- 17075644 TI - Ocular allergic disease: mechanisms, disease sub-types, treatment. AB - Ocular allergy refers to a variety of hypersensitivity disorders that affect the lid, conjunctiva, and/or cornea. Its incidence is estimated at over 20% of the general population in the United States. This review will discuss the various forms of ocular allergy, their pathophysiology, clinical presentation, and treatment. New frontiers in mechanisms, therapy, and management in the office are emphasized throughout. PMID- 17075645 TI - Decoding the packaging insert: indications. PMID- 17075647 TI - The Schirmer test: celebration of its first centenary. PMID- 17075648 TI - Sjogrens syndrome as failed local immunohomeostasis: prospects for cell-based therapy. AB - Sjogrens syndrome has been estimated to affect between 0.2% and 2% or more of the population. It is an autoimmune disease with the hallmark histopathology of focal, periductal, and perivascular CD4(+) cell infiltration of the lacrimal and salivary glands. The immunohistopathology is typically associated with severe lacrimal and salivary dysfunctions, which contribute to debilitating ocular surface and oral symptoms. The quality of life of patients with Sjogrens syndrome often is degraded further by serious, multisystemic manifestations, and they are subject to a forty-fold increased risk of developing B cell lymphomas. In normal lacrimal glands, secretory epithelial cells, autoimmune effector lymphocytes, and regulatory lymphocytes can be seen as collaborating to maintain a local immunohomeostasis. The epithelium contributes by secreting immunomodulatory paracrine factors and also by continuously exposing autoantigens, which thereby become available for uptake by professional antigen presenting cells (APCs). Local or systemic perturbations may initiate autoimmune pathophysiology by impairing the replacement of normally-turning-over regulatory cells, by altering epithelial production of immunomodulatory paracrine factors, by inducing intact epithelial cells to begin secreting previously cryptic epitopes (epitopes that previously were not available to bind to major histocompatibility complex (MHC) molecules and so could not be recognized by T cell antigen receptors), and by inducing epithelial cells to begin expressing MHC Class II molecules and presenting formerly cryptic epitopes directly to CD4(+) cells. This process has been modeled ex vivo with mixed cell reactions comprised of isolated epithelial cells and autologous lymphocytes. This development has occurred as studies of anterior chamber-associated immune deviation (ACAID) and other immunoregulatory phenomena have elucidated the origins and functions of several different kinds of regulatory lymphocytes and shown that regulatory lymphocytes can be generated ex vivo. It now is possible to envision strategies for exploiting each possible mode of epithelial autoantigen exposure to produce therapeutic regulatory cells that might be capable of re-establishing normal immunohomeostasis. Consideration of the hypothetical therapies identifies a number of basic questions that warrant investigation. PMID- 17075649 TI - Drainage of tears: impact on the ocular surface and lacrimal system. AB - The human efferent tear ducts are part of the lacrimal system. Because little knowledge exists concerning the physiology of the nasolacrimal system, and hence its patho- physiology, the nasolacrimal system has received almost no consideration as a possible factor in dry eye. The human nasolacrimal ducts consist of the upper and the lower lacrimal canaliculus, the lacrimal sac, and the nasolacrimal duct. As a draining and secretory system, the efferent tear ducts play a role in tear transport and nonspecific immune defense. Moreover, components of tear fluid are absorbed in the nasolacrimal passage and are transported into a surrounding vascular system. This system is similar to a cavernous body that is subject to autonomic control and regulates tear outflow. Tear duct-associated lymphoid tissue (TALT) is present in the efferent tear ducts, displaying the cytomorphological and immunophenotypic features of mucosa associated lymphoid tissue (MALT). Under normal conditions, tear fluid components are constantly absorbed into the blood vessels of the surrounding cavernous body. These vessels are connected to the blood vessels of the outer eye and could act as a feedback signal for tear fluid production, which ceases if these tear components are not absorbed. In this way, dry eye could be initiated. Defective stimulation of TALT could result in abnormal immune deviation at the ocular surface, leading to an autoimmunological response that causes dry eye pathology. PMID- 17075651 TI - Decoding the package insert: adverse events. PMID- 17075650 TI - Management of the late ocular sequelae of Stevens-Johnson syndrome. AB - The acute conjunctivitis seen initially in Stevens-Johnson syndrome is followed by a cicatricial phase, which often leads to severe ocular surface disease and visual morbidity. Manifestations include keratinization of the conjunctiva, lid margins, and lacrimal and meibomian ducts, resulting in an unstable tear film and mechanical trauma to the conjunctiva and cornea with blinking. Limbal stem cell deficiency is the most vision-threatening sequela of Stevens-Johnson syndrome, as it causes corneal neovascularization, chronic corneal inflammation, and an irregular corneal epithelium. Management of late sequelae often requires a multipronged approach, including strategies for ocular surface protection, ocular surface support, and ocular surface reconstruction. In this review, established therapies, as well as new experimental therapies, are discussed. PMID- 17075652 TI - The Schepens Eye Research Institute, Boston, MA. PMID- 17075654 TI - Fractional Laser treatment for pigmentation and texture improvement. AB - Fractional laser treatment with the 1,550 nm erbium fiber laser (Fraxel Laser, Reliant Technologies) has bridged the gap between the ablative and nonablative laser modalities used to treat the epidermal and dermal signs of skin aging. By targeting water as its chromophore, the laser induces a dense array of microscopic, columnar thermal zones of tissue injury that do not perforate or impair the function of the epidermis. The significant skin remodeling that ensues can be used to treat, with limited downtime, epidermal pigmentation, melasma, and rhytides, as well as textural abnormalities that include acne-related and surgical scars. PMID- 17075653 TI - Topical treatments for melasma and postinflammatory hyperpigmentation. AB - Hyperpigmentation disorders of the skin are common and can be the source of significant psychosocial distress for patients. The most common of these disorders are melasma and postinflammatory hyperpigmentation. Sunscreen use and minimizing sun exposure are crucial in all cases. Topical applications are the mainstay of treatment and include phenols, retinoids, corticosteroids, and their combinations. PMID- 17075655 TI - Tackling Subutex abuse in Singapore. PMID- 17075656 TI - Gregor Mendel (1822-1884): man of God and science. PMID- 17075657 TI - Surgical complications in parenteral Subutex abusers. AB - INTRODUCTION: There were anecdotal reports of an increase in the admissions of parenteral Subutex abusers to our hospital over the past five months. This case study aimed to analyse the surgical complications related to parenteral Subutex abuse and describe the demographics of this group of patients. METHODS: We reviewed all admissions to our hospital between July and November 2005. Only parenteral Subutex abusers were included in this case study. RESULTS: A total of 53 parenteral Subutex abusers were admitted during this period. 31 had surgical complications, while 22 presented with medical ones. Of the surgical patients, 12 had cellulitis and thrombophlebitis, six developed abscesses of the limbs, ten were patients with ischaemia and gangrene of the digits and limbs, one had septic arthritis, one had necrotising fasciitis, and one had a pseudoaneurysm of the femoral artery. There were no reported mortalities. Only nine patients needed surgical interventions. Most of the patients are young with a mean age of 34.2 years. There was a male predominance of 92.4 percent (49 out of 53). Malays are more frequently affected (72 percent, n=38), followed by Indians (15 percent, n=8), and Chinese (13 percent, n=7). CONCLUSION: Parenteral Subutex abuse is a rising concern in Singapore. Many patients present to the surgical and orthopaedic departments for limb and vascular complications. Surgery has a limited role in their management, and most are treated conservatively and expectantly. The solution to this emerging trend requires inter-hospital and ministerial collaboration. PMID- 17075658 TI - Application of artificial neural networks to establish a predictive mortality risk model in children admitted to a paediatric intensive care unit. AB - INTRODUCTION: Paediatric risk of mortality and paediatric index of mortality (PIM) are the commonly-used mortality prediction models (MPM) in children admitted to paediatric intensive care unit (PICU). The current study was undertaken to develop a better MPM using artificial neural network, a domain of artificial intelligence. METHODS: The purpose of this retrospective case series was to compare an artificial neural network (ANN) model and PIM with the observed mortality in a cohort of patients admitted to a five-bed PICU in a Hong Kong non teaching general hospital. The patients were under the age of 17 years and admitted to our PICU from April 2001 to December 2004. Data were collected from each patient admitted to our PICU. All data were randomly allocated to either the training or validation set. The data from the training set were used to construct a series of ANN models. The data from the validation set were used to validate the ANN and PIM models. The accuracy of ANN models and PIM was assessed by area under the receiver operator characteristics (ROC) curve and calibration. RESULTS: All data were randomly allocated to either the training (n=274) or validation set (n=273). Three ANN models were developed using the data from the training set, namely ANN8 (trained with variables required for PIM), ANN9 (trained with variables required for PIM and pre-ICU intubation) and ANN23 (trained with variables required for ANN9 and 14 principal ICU diagnoses). Three ANN models and PIM were used to predict mortality in the validation set. We found that PIM and ANN9 had a high ROC curve (PIM: 0.808, 95 percent confidence interval 0.552 to 1.000, ANN9: 0.957, 95 percent confidence interval 0.915 to 1.000), whereas ANN8 and ANN23 gave a suboptimal area under the ROC curve. ANN8 required only five variables for the calculation of risk, compared with eight for PIM. CONCLUSION: The current study demonstrated the process of predictive mortality risk model development using ANN. Further multicentre studies are required to produce a representative ANN-based mortality prediction model for use in different PICUs. PMID- 17075659 TI - Prevalence and risk factors for iron deficiency in Kelantanese pre-school children. AB - INTRODUCTION: To determine the prevalence of iron deficiency (ID) and iron deficiency anaemia (IDA) in Kelantanese pre-school children and to identify risk factors that best predict the presence of ID. METHODS: Children (aged eight to 26 months) who attended eight primary health clinics in the district of Kota Bharu, Kelantan, Malaysia, from September to November 1999, were invited to participate in the study. Parents were interviewed for potential risk factors of ID and blood was taken for the analysis of haemoglobin (HB), mean corpuscular volume and serum ferritin (SF) level after obtaining a verbal consent. The possible risk factors for ID were compared between the iron deficient (SF less than 12 microgrammes per litre) and iron sufficient (SF greater than 12 microgrammes per litre) groups using multiple logistic regression. RESULTS: Among 490 children, 319 (65.1 percent, 95 percent confidence interval [CI] 60.7- 69.2 percent) had anaemia (HB less than 11.0 g/dL). 191 children (38.9 percent, 95 percent CI 34.7-43.5 percent) had ID. 155 children (31.6 percent, 95 percent CI 27.6-36.0 percent) had IDA (HB less than 11.0 g/dL and SF less than 12 microgrammes per litre). Independent risk factors for ID were prolonged breast feeding for more than six months (p-value is 0.003, adjusted odds ratio [OR] 2.5, 95 percent CI 1.5-4.0) and failure to receive formula milk (p-value is 0.004, adjusted OR 1.6, 95 percent CI 1.2-2.0). Other dietary factors were not significantly associated with ID. CONCLUSION: The prevalence of ID and IDA among Kelantanese children is high, and is mainly attributed to prolonged breast feeding beyond six months of age and failure to introduce formula milk at later infancy. PMID- 17075660 TI - Isolation of monoclonal antibodies-escape variant of dengue virus serotype 1. AB - INTRODUCTION: During an outbreak from December 2004 to March 2005, 138 isolates of dengue virus were prospectively obtained from acute-phase serum samples of 1,067 patients with the provisional clinical diagnosis of acute dengue illness admitted to the adult wards of Hospital Tengku Ampuan Rahimah, Klang, Malaysia. Of the 138 dengue virus isolates, 87, 11, 24 and 3 were typed as dengue serotypes 1, 2, 3 and 4, respectively, by a commercial dengue virus typing kit using monoclonal antibodies (Mab). 13 dengue virus isolates could not be assigned to any specific serotype by serotyping Mab and molecular typing using dengue-type specific molecular typing primer pairs. We report the associated clinical features and limited molecular genetics of this Mab-escape dengue virus variant. METHODS: Limited molecular characterisation of the Mab-escape dengue virus variants with respect to a few concurrently isolated dengue serotype 1 virus was performed by reverse transcriptase polymerase chain reaction (RT-PCR), followed by nucleic acid sequencing of the 500-bp dengue virus partial genomic capsid-PreM fragment. RESULTS: The aligned nucleic acid sequence of RT-PCR products showed that these Mab-escape variants were of identical nucleic acid sequence, and shared the highest sequence homology (99 percent) with dengue virus serotype 1 (GeneBank accession No. AB178040) isolated from a Japanese patient in 2004. Though these Mab-escape dengue virus variants were noted to replicate to a 2-log higher titre than the current circulating dengue virus serotype 1, there was no significant difference between these variants and the currently circulating dengue virus serotype 1 with respect to disease severity (dengue fever versus dengue haemorrhagic fever) and clinical features. CONCLUSION: There was no significant difference in the proportion of patients developing dengue haemorrhagic fever following acute infection by Mab-escape dengue virus 1 variant in comparison with infection by the conventional dengue virus 1. Similarly, there was no significant difference in the pattern of clinical presentations following acute infection by the two different strains of virus. PMID- 17075661 TI - Evaluation of bone mineral density in thyrotoxicosis. AB - INTRODUCTION: This project aimed to study the incidence and profile of bone involvement in thyrotoxicosis patients by dual energy X-ray absorptiometry (DEXA) scan and the effect of treatment on the bone mineral density (BMD). METHODS: A total of 50 young patients with a mean age of 29.4 years, diagnosed to have thyrotoxicosis clinically and proven by thyroid function tests, were included in this prospective three-year study conducted at the Madras Medical College and Government General Hospital in Chennai, India. Patients were enrolled if they had bone pain or had elevation of serum alkaline phosphatase. All these patients had a baseline BMD measurement by DEXA scans in the region of the lumbar vertebrae before treatment and the T-score was computed. All other secondary causes of low BMD, like primary hyperparathyroidism, long-term steroid intake, vitamin D deficiency, was ruled out. After definitive management of hyperthyroidism by anti thyroid drugs and surgery, all the patients with bone involvement had a repeat DEXA scan after one year and the T-score was computed. RESULTS: Out of 50 patients, 46 had bone involvement (92 percent). Based on the World Health Organisation classification, 16 (32 percent) had osteopenia and 30 patients (60 percent) had osteoporosis. After control of thyrotoxicosis, the mean bone mass increased from 0.729 g/sq cm to 0.773 g/sq cm, a statistically significant increase of 0.044 g/sq cm (p-value is less than 0.001) after one year, compared to age- and sex-matched controls. The mean percentage of the bone mass compared to the peak BMD increased from 70.2 percent to 74.2 percent after treatment, an increase of four percent (p-value is less than 0.001). The mean percentage of the bone mass compared to the age-matched BMD increased from 71.2 percent to 75.2 percent after treatment, an increase of four percent (p-value is less than 0.001), all of which were statistically significant. CONCLUSION: Metabolic bone disease should be looked for in all thyrotoxic patients, especially patients complaining of bone pain and those with elevated bone enzymes. DEXA scans offer a convenient, reliable and noninvasive modality for diagnosis and monitoring therapy. PMID- 17075662 TI - Computer-integrated patient-controlled epidural analgesia: a preliminary study on a novel approach of providing pain relief in labour. AB - INTRODUCTION: The need for individualisation of analgesic therapy in labour cannot be overemphasised. We have devised a programme, based on a novel clinical algorithm, that converts a continuous infusion pump into a patient-controlled epidural analgesia (PCEA) pump that is responsive to the patient's needs by varying its rate of infusion. METHODS: In this double-blinded, controlled trial, 40 American Society of Anesthesiologists 1 patients were recruited to receive either a continuous infusion of 10 ml/hour (Continuous Epidural Infusion [CEI], n=20) or the computer-integrated (CI) regimen (CI-PCEA, n=20) to maintain epidural analgesia after successful induction of combined spinal analgesia during early labour. The proportion of patients who had delivered without a requirement for analgesic supplementation was the primary outcome measure. RESULTS: There was a significant difference in the incidence of breakthrough pain, i.e. the primary outcome measure (two in CI-PCEA versus eight in the CEI group, p-value is 0.027). There was a trend towards a longer duration before analgesic supplementation of analgesia was required after its induction with CI-PCEA than CEI (p-value is 0.06). We could not detect a difference in the total hourly consumption of epidural analgesics between the two groups. CONCLUSION: Our study also showed that with the CI-PCEA programme, we were able to convert an ordinary infusion pump to one which analyses the patients' needs in the previous hour (based on analgesic demands) and automatically adjusts the basal infusion accordingly. CI PCEA reduced the incidence of breakthrough pain without the evidence of increasing drug consumption when compared with CEI. PMID- 17075663 TI - Co-sleeping and clinical correlates in children seen at a child guidance clinic. AB - INTRODUCTION: Co-sleeping or bed-sharing is a common practice that has been little researched. While often viewed as being "cultural" in nature, there is a suggestion that it may be a parental response to sleep problems. Some studies link co-sleeping with behavioural and temperamental difficulties. The objectives of the current study were to determine the prevalence of co-sleeping and how they relate to sleeping problems among a cohort of children and adolescents seen in a child guidance clinic. METHODS: Parents or guardians of all new patients seen at the child guidance clinic were asked to complete a questionnaire upon their consent to participate in the study. The questionnaire included socio demographical data and frequency of sleep problems in the past six months. A list of nine common sleep problems was included. RESULTS: The prevalence of co sleeping was found to be 72.7 percent. The children who co-sleep were significantly younger and there was a decrease in the practice with increasing age. Sleep starts and nightmares were significantly more among those sleeping alone. CONCLUSION: Co-sleeping was not associated with significant sleep problems in our cohort. Co-sleeping may have been initiated in response to an existing sleep problem but eventually resolved the problem. If co-sleeping is not permitted, the sleep problem could be compounded, giving rise to a higher prevalence of sleep starts and nightmares among those in our cohort who slept alone. PMID- 17075664 TI - Positive health practices and depressive symptoms among high school adolescents in Oman. AB - INTRODUCTION: The study aimed to investigate the association of health practices and depressive symptoms among high school adolescents in a national representative sample of 5,409 students in Oman. METHODS: Depressive symptoms were screened in 2004 through the application of the self-report 27-item Child Depression Inventory. Health practices scale comprised a simple sum of five healthy practices, namely: sleeping seven to eight hours at night, having breakfast daily, not eating between meals, not smoking the month prior to the study, and doing physical activities more than once per week apart from attending physical education classes in school. RESULTS: Sequential logistic regression models were run to test for the change in the odds-ratio (OR) of having depressive illness with a one point increase in the healthy practices scale, after adjustment for other risk factors of depression. Health practices remained having a significant protective effect on depression (OR is 0.72, 95 percent confidence interval is 0.64-0.80) even after adjustment to other significant covariates in the last model, such as history of chronic medical or mental illness diagnosed by a doctor, high scoring in chance health locus of control (HLC), low scoring in internal HLC, poor relationships with social contacts, and physical abuse during childhood or adolescence. CONCLUSION: Findings support the protective effects of positive health practices on adolescents' depression. PMID- 17075665 TI - Measurement of cardiac output using Physio Flow with different positions of electrode placement. AB - INTRODUCTION: Physio Flow is a non-invasive impedance cardiograph device that measures cardiac output. Recommended electrode placements involve six electrodes, including two near the xiphisternum (Z3 and Z4/ ECG3/neutral). This study aims to evaluate if changing the positions of these two leads to the left fourth and fifth intercostal spaces along the mid-axillary line results in a change in the cardiac output measurement. METHODS: This was a prospective, controlled, crossover, paired study of 30 patients where electrodes were placed in the recommended positions and cardiac output (CO1) obtained after two minutes. The second cardiac output (CO2) was then obtained with the electrodes Z3 and Z4/ECG3/neutral repositioned at the left mid-axillary line at the fourth and fifth intercostal spaces. The final step involved switching the Z3 and Z4/ECG3/neutral leads back to the recommended position and the cardiac output (CO3) was measured. RESULTS: The average of the initial and third readings (COave) was compared with the measured CO2 and analysed. The regression equation was: CO at the proposed site (CO2) = COave at the recommended site + 0.058. The paired samples correlation was 0.995. Within the 95 percent limits of agreement, the bias with CO measured at the proposed site of electrode placement was 0.046 L/min with the limits at -0.24 L/min and 0.34 L/min. The mean difference was 0.86% of the average CO. CONCLUSION: A small positive bias was demonstrated when Physio Flow measurements were taken with the leads Z3 and Z4/ECG3/neutral placed in the mid-axillary line fourth and fifth intercostal spaces. PMID- 17075666 TI - Intrapartum maternal sinus bradycardia with spontaneous resolution following delivery. AB - Pregnancy is a normal physiological state that is characterised by significant adaptive changes, many of which take place in the cardiovascular system. These adaptations are believed to be the cause for the increased incidence in cardiac arrhythmias in pregnancy and particularly, in labour. We report an unusual case of a healthy 32-year-old primigravida who developed profound intrapartum bradycardia which persisted throughout labour. Spontaneous recovery to pre-labour baseline heart rate occurred following spontaneous vaginal delivery. Maternal and foetal statuses remained satisfactory during labour. This case report underscores the importance of recognising that arrhythmias in various forms are common in labour. In the absence of detectable underlying cardiac disease or maternal and foetal compromise, continuation of the labour with a view to vaginal delivery should be the goal. PMID- 17075667 TI - Dengue encephalitis: why we need to identify this entity in a dengue-prone region. AB - Classical dengue fever is commonly seen in children and young adults. It commonly presents with fever, severe headache, body ache and retro-orbital pain. Unlike other arboviral infections, dengue virus does not usually cause neurological manifestations. We report a 13-year-old boy with dengue encephalitis. Dengue encephalitis should be considered in the differential diagnosis of acute viral encephalitis especially in countries like India where dengue has assumed epidemic proportions. These undiagnosed cases are at risk of developing complications of dengue haemorrhagic fever. PMID- 17075668 TI - Massive unilateral chylous pleural effusion: a rare initial presentation of Behcet's disease. AB - Pulmonary manifestations of Behcet's disease are not very common and usually include pulmonary artery aneurysms, central venous thrombosis, pneumonia and pleurisy. Chylothorax secondary to superior vena caval obstruction is a rare complication and has been reported in only a few cases. We report a case of a 24 year-old man presenting with massive chylothorax as the initial presentation of Behcet's disease that was successfully treated conservatively. PMID- 17075669 TI - Phenytoin-induced parkinsonism. AB - This report presents a 30-year-old man who developed subacute phenytoin-induced cerebellar ataxia and parkinsonism that resolved after discontinuation of the phenytoin treatment. Phenytoin was started for seizure prophylaxis in another health institution where he was referred for bilateral intracerebral orbitofrontal haemorrhage due to a head trauma. To our knowledge, there has been only one other case report describing phenytoin-induced parkinsonism, which was also reversible. The issue of the development of parkinsonism due to the phenytoin toxicity in the case of bilateral orbitofrontal lesion is addressed. PMID- 17075671 TI - Clinics in diagnostic imaging (112). Perinatal lethal hypophosphatasia (PLH). AB - A two-hour-old female infant presented with respiratory distress and short limbs. Neonatal radiographs showed micromelic dwarfism and generalised demineralisation, especially at the ribs, long bones of both forearms and both fibulae. The spine showed a flattened shape. All long bones showed metaphyseal irregularities and flaring. Normal serum calcium and elevated serum phosphorus were found, while serum alkaline phosphatase was markedly reduced. A diagnosis of perinatal lethal hypophosphatasia was made. The aetiology, clinical manifestations, radiographical findings, laboratory assays, prenatal diagnosis and treatment of hypophosphatasia are discussed. PMID- 17075670 TI - Intrapleural fibrinolysis in clotted haemothorax. AB - The immediate treatment of haemothorax is thoracostomy tube insertion. One complication of haemothorax is retained blood and if improperly managed, this can lead to complications such as empyema and fibrothorax. The ideal management of clotted haemothorax is a matter of controversy. Video-assisted thoracoscopic surgery (VATS) is believed to be the best available modality for the management of clotted haemothorax. However, VATS is not routinely available in many centres. One easily available and effective alternative to VATS is the use of intrapleural fibrinolysis. We report the successful management of a post-traumatic clotted haemaothorax in a 34-year-old man, using intrapleural instillation of streptokinase, and review the literature on the management of clotted haemothorax. PMID- 17075672 TI - Will "no blood" kill Jehovah Witnesses? AB - A 46-year-old Indonesian woman presented with signs and symptoms suggestive of an ovarian tumour and was advised to have surgery with exploratory laparotomy and removal of the mass. She agreed but refused blood transfusion any time in the course of her treatment or procedure, as she was a Jehovah Witness. As there was a high risk of intraoperative haemorrhage, steps were taken to reduce any consequent complications due to the surgery. The ethical conflict is between respecting patient autonomy and compromising standards of care, arising from the refusal of a standard therapy. The latest developments in the blood transfusion doctrine policy for the Jehovah Witnesses are also discussed in this case study. PMID- 17075673 TI - Mothballs: bringing safety issues out from the closet. PMID- 17075674 TI - Acute undifferentiated leukaemia in adult presenting as arthritis. PMID- 17075675 TI - Simulataneous bilateral occulomotor nerve paralysis: an unusual manifestation of diabetes mellitus. PMID- 17075676 TI - Self-inflating resuscitators for assisted respiration in chest trauma: the hidden risk. PMID- 17075677 TI - Fatal bacteraemic pneumonia due to community-acquired methicillin-resistant Staphylococcus aureus. PMID- 17075679 TI - Knowledge that upper respiratory tract infection resolves on its own is associated with more appropriate health-seeking behaviour and antibiotic cognition. PMID- 17075680 TI - Predicting the outcome of percutaneous endoscopic gastrostomy tube feeding. PMID- 17075681 TI - Can chlorpheniramine cause serotonin syndrome? PMID- 17075684 TI - Side errors in neurosurgery. AB - Eight cases of neurosurgical operations being performed on the wrong side were studied. Safeguards of confidentiality were used. In seven cases side marking was not done and the surgeons felt that had it been done, the mistake would have or may have been prevented. In all but one case the surgeon's normal side check was omitted. Distracting circumstances contributed to the checks being omitted in these seven cases. In the one case where the check was carried out the mistake was made because the patient had been marked on the wrong side. In no case was the mistake made because of ambiguous or absent site data in the imaging or notes. It is concluded that to prevent these mistakes emphasis should be placed on ensuring that the preoperative site check is completed more than on ensuring that unambiguous side information is available in the notes and imaging. PMID- 17075685 TI - Cloning of the black seabream (Acanthopagrus schlegeli) antiquitin gene and functional characterization of its promoter region. AB - Antiquitin (ALDH7) is a member of the aldehyde dehydrogenase superfamily. In plants, ALDH7 is inducible upon dehydration and is thus believed to possess an osmoregulatory role. On the other hand, however, its exact physiological function in animals remains elusive. We herein report the isolation of the black seabream (Acanthopagrus schlegeli) antiquitin gene (sbALDH7) and the functional characterization of its promoter region. The 1.6 kb 5'-flanking region of sbALDH7 exhibits an intense promoter activity (30-170 fold of the basal) in five mammalian and fish cell lines of different origins. Progressive 5'-deletion analysis suggests that the core promoter is located within the region -297/+41 whereas a cis-acting repressor of basal transcription is present in the region 878/-297. In silico analysis of this sbALDH7 promoter region does not reveal any osmotic response element. Instead, it contains potential binding sites for cell cycle related cis-elements such as CCAAT displacement protein and cell cycle dependent element/cell cycle genes homology region. PMID- 17075686 TI - Purification and properties of glutathione reductase from liver of the anoxia tolerant turtle, Trachemys scripta elegans. AB - Glutathione reductase (GR) is a homodimeric flavoprotein that catalyzes the reduction of oxidized glutathione (GSSG) using NADPH as a cofactor. The enzyme is a major component of cellular defense mechanisms against oxidative injury. In this study, GR was purified from the liver of the anoxia-tolerant turtle, Trachemys scripta elegans. The overall fold purifications were 13.3- and 12.1 fold with final specific activities of 5.5 and 1.44 U/mg of protein for control and anoxic turtle GR, respectively. SDS-PAGE of purified turtle liver GR showed a single protein band at approximately 55 kDa. Reverse phase HPLC of turtle GR revealed a single peak that had the same retention time as yeast GR. No new isoform of GR was detected in liver of T. s. elegans during anoxia. The K (m) values of turtle GR for GSSG and NADPH was 44.6 and 6.82 microM, respectively, suggesting a substantially higher affinity of turtle GR toward GSSG than most other vertebrates. Unlike other human GR, NADP(+ )did not inhibit turtle GR activity. The activation energy of turtle GR, calculated from the slope of the Arrhenius plot, was 32.2 +/- 2.64 kJ/mol. Turtle GR had high activity under a broad pH range (having activity between pHs 4 and 10; optimal activity at pH 6.5) and the enzyme maintains activity under the pH drop that occurs under anoxic conditions. The high affinity of turtle GR suggests that turtles have high redox buffering capacity of tissues to protect against oxidative stress encountered during anoxia/reoxygenation. PMID- 17075687 TI - Current and future use of positron emission tomography (PET) in breast cancer. AB - Positron emission tomography (PET) is a radiotracer imaging method that is increasingly used in both the clinical care of breast cancer patients and in translational breast cancer research. This review emphasizes current and future clinical applications of PET to breast cancer, and highlights some translational research using PET to elucidate the clinical biology of breast cancer. PET principles are reviewed, followed by a review of current applications of (18)F fluorodeoxyglucose (FDG) to clinical breast cancer care. Finally we review work done with other radiopharmaceuticals beyond FDG designed to image a number of aspects of breast cancer biology, emphasizing those most likely to enter clinical trials in the near future. PMID- 17075688 TI - Management of psychiatric disorders in children and adolescents with atypical antipsychotics: a systematic review of published clinical trials. AB - We aimed to provide a descriptive review of treatment studies of atypical antipsychotics in paediatric psychiatric disorders. A systematic review of the literature used Medline and EMBASE databases to identify clinical trials of atypical antipsychotics in children and adolescents between 1994 and 2006. Trials were limited to double-blind studies and open-label studies of > or = 8 weeks duration that included > or = 20 patients. Nineteen double-blind and 22 open label studies were identified. Studies included use of clozapine, olanzapine, quetiapine, risperidone, and ziprasidone in the treatment of disruptive behavioural disorders (DBDs), pervasive developmental disorders (PDDs), tic disorder, psychotic disorders, and mania. These medications generally reduced the severity of a variety of psychiatric symptoms in children and adolescents. Less frequent adverse events included extrapyramidal symptoms, hyperglycaemia and diabetes, and endocrine effects. The review of published scientific data suggests that most of the atypical antipsychotics, excluding clozapine, have a favourable risk/benefit profile and effectively reduce disabling behaviours in paediatric psychiatric patients. While there is a body of evidence published of treatment of DBDs and PDDs, there is a lack of controlled data to guide clinical practice for the use of atypical antipsychotics for paediatric psychotic disorders and bipolar disorder. While there have been studies with duration up to 2 years, no definitive data are available that suggest long-term safety; additional studies are warranted. PMID- 17075689 TI - Brain bank of the Brazilian aging brain study group - a milestone reached and more than 1,600 collected brains. AB - INTRODUCTION: Brain banking remains a necessity for the study of aging brain processes and related neurodegenerative diseases. In the present paper, we report the methods applied at and the first results of the Brain Bank of the Brazilian Aging Brain Study Group (BBBABSG) which has two main aims: (1) To collect a large number of brains of elderly comprising non-demented subjects and a large spectrum of pathologies related to aging brain processes, (2) To provide quality material to a multidisciplinar research network unraveling multiple aspects of aging brain processes and related neurodegenerative diseases. METHODS: The subjects are selected from the Sao Paulo Autopsy Service. Brain parts are frozen and fixated. CSF, carotids, kidney, heart and blood are also collected and DNA is extracted. The neuropathological examinations are carried out based on accepted criteria, using immunohistochemistry. Functional status are assessed through a collateral source based on a clinical protocol. Protocols are approved by the local ethics committee and a written informed consent form is obtained. RESULTS: During the first 21 months, 1,602 samples were collected and were classified by Clinical Dementia Rating as CDR0: 65.7%; CDR0.5:12.6%, CDR1:8.2%, CDR2:5.4%, and CDR3:8.1%. On average, the cost for the processing each case stood at 400 US dollars. To date, 14 laboratories have been benefited by the BBBABSG. CONCLUSION: The high percentage of non- demented subjects and the ethnic diversity of this series may be significantly contributive toward aging brain processes and related neurodegenerative diseases understanding since BBBABSG outcomes may provide investigators the answers to some additional questions. PMID- 17075690 TI - Students learning from patients: let's get real in medical education. AB - Medical students must be prepared for working in inter-professional and multi disciplinary clinical teams centred on a patient's care pathway. While there has been a good deal of rhetoric surrounding patient-centred medical education, there has been little attempt to conceptualise such a practice beyond the level of describing education of communication skills and empathy within a broad 'professionalism' framework. Paradoxically, while aiming to strengthen patient student interactions, this approach tends to refocus on the role modelling of the physician, and opportunities for potentially deep collaborative working relationships between students and patients are missed. A radical overhaul of conventional doctor-led medical education may be necessary, that also challenges the orthodoxies of individualistic student-centred approaches, leading to an authentic patient-centred model that shifts the locus of learning from the relationship between doctor as educator and student to the relationship between patient and student, with expert doctor as resource. Drawing on contemporary poststructuralist theory of text and identity construction, and on innovative models of work-based learning, the potential quality of relationship between student and patient is articulated in terms of collaborative knowledge production, involving close reading with the patient as text, through dialogue. Here, a medical 'education' displaces traditional forms of medical 'training' that typically involve individual information reproduction. Students may, paradoxically, improve clinical acumen through consideration of silences, gaps, and contradictions in patients as texts, rather than treating communication as transparent. Such paradoxical effects have been systematically occluded or denied in traditional medical education. PMID- 17075691 TI - Mutation and haplotype analyses of the MUT gene in Japanese patients with methylmalonic acidemia. AB - Methylmalonic acidemia (MMA) is caused by a deficiency in the activity of L: methylmalonyl-CoA mutase (MCM), a vitamin B12 (or cobalamin, Cbl)-dependent enzyme. Apoenzyme-deficient MMA (mut MMA) results from mutations in the nuclear gene MUT. Most of the MUT mutations are thought to be private or restricted to only a few pedigrees. Our group elucidated the spectrum of mutations of Japanese mut MMA patients by performing mutation and haplotype analyses in 29 patients with mut MMA. A sequence analysis identified mutations in 95% (55/58) of the disease alleles. Five mutations were relatively frequent (p.E117X, c.385 + 5G > A, p.R369H, p.L494X, and p.R727X) and four were novel (p.M1V, c.753_753 + 5delGGTATA, c.1560G > C, and c.2098_2099delAT). Haplotype analysis suggested that all of the frequent mutations, with the exception of p.R369H, were spread by the founder effect. Among 46 Japanese patients investigated in the present and previous studies, 76% (70/92) of the mutations were located in exons 2, 6, 8, and 13. This finding - that a limited number of mutations account for most of the mutations in Japanese mut MMA patients - is in contrast with results of a previous study in Caucasian patients. PMID- 17075692 TI - Alpha-adrenoceptor gene variants and autonomic nervous system function in a young healthy Japanese population. AB - alpha(1A)-adrenergic receptor (alpha(1A)-AR) regulates the cardiac and peripheral vascular system through sympathetic activation, and alpha(2A)-AR and alpha(2C)-AR subtypes are essential for presynaptic feedback regulation of catecholamine release from the central and peripheral sympathetic nerve. Genetic variations in each human alpha-AR subtype gene have been identified and have been implicated in hypertension and cardiovascular disease. It is not yet clear whether these genetic variations actually have an effect on sympatho-vagal modulation. The aim of the present study was to evaluate the relation between the five representative genetic polymorphisms of alpha-AR subtypes (Arg347Cys of alpha(1A)-AR; C-1291G, Asn251Lys, and DraI RFLP of alpha(2A)-AR; and Del322-325 of alpha(2C)-AR) and autonomic nervous system (ANS) function in young and healthy Japanese males. One hundred forty-nine subjects were genotyped for each alpha-AR polymorphism, and underwent evaluation of ANS function by power spectral analysis of heart rate variability (HRV) during supine rest and in a standing position. In a supine position, the alpha(1A)-AR 347Cys allele was significantly associated with lower HRV sympathetic index (normalized low frequency power [LF(%)] and LF:HF ratio) and higher HRV parasympathetic index [HF(%)]. Meanwhile, subjects with the alpha(2C)-AR Del322-325 allele had markedly higher LF(%) and LF:HF ratio and lower HF(%) than noncarriers. Thus, the alpha(1A)-AR and alpha(2C)-AR genetic variations influence sympatho-vagal balance even in young and healthy normotensive states, which could be postulated to constitute an intermediate phenotype for future pathological episodes of various ANS dysfunction-related diseases. PMID- 17075693 TI - Optimization of the cellular import of functionally active SH2-domain-interacting phosphopeptides. AB - Phosphopeptides interacting with src homology 2 (SH2) domains can activate essential signaling enzymes in vitro. When delivered to cells, they may disrupt protein-protein interactions, thereby influencing intracellular signaling. We showed earlier that phosphopeptides corresponding to the inhibitory motif of Fcgamma receptor IIb and a motif of the Grb2-associated binder 1 adaptor protein activate SH2-containing tyrosine phosphatase 2 in vitro. To study the ex vivo effects of these peptides, we have now compared different methods for peptide delivery: (i) permeabilization of the target cells and (ii) the use of cell permeable vectors, which are potentially able to transport biologically active compounds into B cells. We found octanoyl-Arg(8) to be an optimal carrier for the delivery of phosphopeptides to the cells. With this strategy, the function of cell-permeable SHP-2-binding phosphopeptides was analyzed. These peptides modulated the protein phosphorylation in B cells in a dose- and time-dependent manner. PMID- 17075694 TI - L-arginine regulates asymmetric dimethylarginine metabolism by inhibiting dimethylarginine dimethylaminohydrolase activity in hepatic (HepG2) cells. AB - An increase in circulating asymmetric dimethylarginine (ADMA) and a decreased L arginine/ADMA ratio are associated with reduced endothelial nitric oxide (NO) production and increased risk of vascular disease. We explored relations between ADMA, L-arginine and dimethylarginine dimethylaminohydrolase (DDAH) in liver (HepG2) cells. DDAH is the principle enzyme for the metabolism of ADMA. HepG2 cells metabolised 44.8 nmol/h of ADMA per 3.6 x 10(6) cells in the absence of L arginine. The metabolism of ADMA at physiological (1mu mol/l, p < 0.01) and at pathological (100mumol/l, p < 0.01) levels was inhibited dose-dependently by L arginine (0-400mumol/l) in cultured HepG2 cells and increased intracellular ADMA (p = 0.039). L-arginine competitively inhibited DDAH enzyme activity to 5.6 +/- 2.0% of the untreated level (p < 0.01). We conclude that L-arginine regulates ADMA metabolism dose-dependently by competing for DDAH thus maintaining the metabolic balance of L-arginine and ADMA, and endothelial NO homeostasis. PMID- 17075695 TI - Chitotriosidase: the yin and yang. AB - The enzyme chitotriosidase (ChT), the human analogue of chitinases from non vertebrate species, is one of the most abundant and indicative proteins secreted by activated macrophages. Its enzymatic activity is elevated in serum of patients suffering from Gaucher's disease type 1 and in some other inherited lysosomal storage disorders, as well as in diseases in which macrophages are activated. The last decade has witnessed the appearance of a substantial number of studies attempting to unravel its cellular functions, which have yet not been fully defined. A great deal of progress has been made in the study of the physiological roles of ChT. This review is looks at the key areas of investigations addressed to further illuminate whether ChT activation might have different functional meanings in various diseases. PMID- 17075696 TI - Immunophilins: for the love of proteins. AB - Immunophilins are chaperones that may also exhibit peptidylprolyl isomerase (PPIase) activity. This review summarizes our knowledge of the two largest families of immunophilins, namely cyclophilin and FK506-binding protein, and a novel chimeric dual-family immunophilin, named FK506- and cyclosporin-binding protein (FCBP). The larger members of each family are modular in nature, consisting of multiple PPIase and/or protein-protein interaction domains. Despite the apparent difference in their sequence and three-dimensional structure, the three families encode similar enzymatic and biological functions. Recent studies have revealed that many immunophilins possess a chaperone function independent of PPIase activity. Knockout animal studies have confirmed multiple essential roles of immunophilins in physiology and development. An immunophilin is indeed a natural 'protein-philin' (Greek 'philin' = friend) that interacts with proteins to guide their proper folding and assembly. PMID- 17075697 TI - Comparative genomic analysis of human and chimpanzee indicates a key role for indels in primate evolution. AB - Sequence comparison of humans and chimpanzees is of interest to understand the mechanisms behind primate evolution. Here we present an independent analysis of human chromosome 21 and the high-quality BAC clone sequences of the homologous chimpanzee chromosome 22. In contrast to previous studies, we have used global alignment methods and Ensembl predictions of protein coding genes (n = 224) for the analysis. Divergence due to insertions and deletions (indels) along with substitutions was examined separately for different genomic features (coding, noncoding genic, and intergenic sequence). The major part of the genomic divergence could be attributed to indels (5.07%), while the nucleotide divergence was estimated as 1.52%. Thus the total divergence was estimated as 6.58%. When excluding repeats and low-complexity DNA the total divergence decreased to 2.37%. The chromosomal distribution of nucleotide substitutions and indel events was significantly correlated. To further examine the role of indels in primate evolution we focused on coding sequences. Indels were found within the coding sequence of 13% of the genes and approximately half of the indels have not been reported previously. In 5% of the chimpanzee genes, indels or substitutions caused premature stop codons that rendered the affected transcripts nonfunctional. Taken together, our findings demonstrate that indels comprise the majority of the genomic divergence. Furthermore, indels occur frequently in coding sequences. Our results thereby support the hypothesis that indels may have a key role in primate evolution. PMID- 17075698 TI - Local mutagenic impact of insertions of LTR retrotransposons on the mouse genome. AB - Solitary LTR loci are the predominant form of LTR retrotransposons in most eukaryotic genomes. They originate from recombination between the two LTRs of an ancestral retrovirus and are therefore incapable of transposition. Despite this inactivity, they appear to have a substantial impact on the host genome. Here we use the murine RMER10 LTR family as an example to describe how such elements can reshape regions of the genome through multiple mutations on an evolutionary time scale. Specifically, we use phylogenetic analysis of multiple copies of RMER10 in rodent species, as well as comparisons of orthologous pairs in mouse and rat, to argue that insertions of members of this family have locally induced the emergence of tandem repeat loci as well as many indels. Analysis of structural aspects of these sequences (secondary structures and transcription factors signals) may explain why RMER10 can become endogenous "mutagenic" factors through induction of replication fork blockages and/or error-prone repair of aberrant DNA structures. This hypothesis is also consistent with features of other interspersed repeated elements. PMID- 17075699 TI - Mitochondrial gene rearrangements and partial genome duplications detected by multigene asymmetric compositional bias analysis. AB - Asymmetric compositional and mutation bias between the two strands occurs in mitochondrial genomes, and an asymmetric mechanism of mtDNA replication is a potential source of this bias. Some evidence indicates that during replication the heavy strand is subject to a gradient of time spent in a single-stranded state (D (ssH)) and a gradient of mutational damage. The nucleotide composition bias among genes varies with D (ssH). Consequently, partial genome duplications (PGD) will alter the skew for genes located downstream of the duplication, relatively to nascent light strand synthesis, and in the same way, gene rearrangements (GRr) will affect genes by changing their skews. We examined cases where there had been PGD or GRr and determined whether this left a trace in the form of unusual patterns of base composition. We compared the skew of genes differently located on the mtDNA genome of previously published whole mtDNA genomes from amphibians, a group that shows considerable levels of both GRr and PGD. After observing a significant correlation between AT and GC skew with D (ssH) at fourfold redundant sites, we ran our analysis and detected 31.3% of the species with GRr and/or PGD. By comparing the nucleotide composition at fourfold redundant sites in normal and "abnormal" species, we found that A/C variation occurs and is associated with GRr/PGD. These results show that by analyzing the nucleotide skews of only three genes, it may be possible to predict some mitochondrial GRr and/or PGD without knowing the complete mtDNA genome sequence. PMID- 17075700 TI - Protein modules conserved since LUCA. AB - Universal scale of the sequence conservation has been recently introduced based on omnipresence of the protein sequence motifs across species. A large spectrum of short sequences, up to eight residues has been found to reside in all or almost all prokaryotic organisms. By this discovery a principally novel quantitative approach is introduced to the problem of reconstruction of the last universal common ancestor (LUCA). The most conserved elements (protein modules) with defined structures and sequences harboring the omnipresent motifs are outlined in this work, by combining the sequence and protein crystal structure data. The structurally conserved modules involve 25-30 amino acid residues and have appearance of closed loops, loop-n-lock structures. This confirms earlier conclusions on the loop-fold structure of globular proteins. Many of the topmost conserved modules represent the primary closed loop prototypes, that have been derived by whole genome sequence searches. The data presented, thus, make a basis for further developments toward the earliest stages of protein evolution. PMID- 17075701 TI - Electrophysiological validation of a human prototype auditory midbrain implant in a guinea pig model. AB - The auditory midbrain implant (AMI) is a new treatment for hearing restoration in patients with neural deafness or surgically inaccessible cochleae who cannot benefit from cochlear implants (CI). This includes neurofibromatosis type II (NF2) patients who, due to development and/or removal of vestibular schwannomas, usually experience complete damage of their auditory nerves. Although the auditory brainstem implant (ABI) provides sound awareness and aids lip-reading capabilities for these NF2 patients, it generally only achieves hearing performance levels comparable with a single-channel CI. In collaboration with Cochlear Ltd. (Lane Cove, Australia), we developed a human prototype AMI, which is designed for electrical stimulation along the well-defined tonotopic gradient of the inferior colliculus central nucleus (ICC). Considering that better speech perception and hearing performance has been correlated with a greater number of discriminable frequency channels of information available, the ability of the AMI to effectively activate discrete frequency regions within the ICC may enable better hearing performance than achieved by the ABI. Therefore, the goal of this study was to investigate if our AMI array could achieve low-threshold, frequency specific activation within the ICC, and whether the levels for ICC activation via AMI stimulation were within safe limits for human application. We electrically stimulated different frequency regions within the ICC via the AMI array and recorded the corresponding neural activity in the primary auditory cortex (A1) using a multisite silicon probe in ketamine-anesthetized guinea pigs. Based on our results, AMI stimulation achieves lower thresholds and more localized, frequency-specific activation than CI stimulation. Furthermore, AMI stimulation achieves cortical activation with current levels that are within safe limits for central nervous system stimulation. This study confirms that our AMI design is sufficient for ensuring safe and effective activation of the ICC, and warrants further studies to translate the AMI into clinical application. PMID- 17075702 TI - CC chemokine receptor 2 is protective against noise-induced hair cell death: studies in CX3CR1(+/GFP) mice. AB - Acoustic trauma was recently shown to induce an inflammatory response in the ear characterized by rapid entry of macrophages in the spiral ligament. The current study seeks to elucidate the mechanisms involved in summoning macrophages to the cochlear lateral wall and the role macrophages play in noise-induced injury or repair. CCL2 and its primary receptor, CCR2, are the most widely validated effectors of monocyte chemotaxis in vivo. CCL2-/- and CCR2-/- mice have been used extensively in studies of monocyte activation in neuronal injury. However, the function of CCL2 and CCR2 in the cochlea has not been studied. The present study examines the role of CCL2 and CCR2 in acoustic injury. CCL2-/- and CCR2-/- mice on a CX3CR1(+/GFP) background were exposed to octave band noise (8-16 kHz) for 2 h to determine the effect of CCL2 and CCR2 on monocyte migration into the cochlea, threshold shift, and cell survival. We found that threshold shift was unchanged in the two knockout mouse strains when compared to the background strain (CX3CR1(+/GFP)). Surprisingly, we found that monocyte migration was also unchanged, despite the absence of CCL2 or CCR2. However, there was a dramatic increase in noise-induced hair cell death in the CCR2-/- strain. This observation suggests that CCR2, independent of CCL2, plays a protective role in the cochlea after noise, and neither ligand nor receptor is necessary for monocyte migration. Possible mechanisms of neuroprotection by CCR2 are discussed. PMID- 17075703 TI - Phylogenetic analyses within three sections of the genus Vicia. AB - The averaged genomic similarities based on multilocus randomly amplified polymorphic DNA (RAPD) were calculated for eight species representing three sections of the genus Vicia: faba, bithynica and narbonensis. The frequency of appearance of the sequences corresponding to 25 decamers selected at random from genomes of different Fabace species was checked, and a high correlation with the frequency observed for Vicia allowed us to assume their similar weight in typing Vicia species. The RAPD-based similarity coefficients compared with those related to whole genome hybridization with barley rDNA and those based on restriction fragment length polymorphism (RFLP) revealed similar interspecies relationships. The averaged RAPD-based similarity coefficient (Pearson's) was 0.68 for all the species, and was sectionspecific: 0.43 (bithynica), 0.50 (faba) and 0.73 (narbonensis). The averaged similarity coefficient for V. serratifolia (0.63) placed it apart from the rest (0.75) of its section. The results correspond to the interspecies relationships built upon non-genetic data. The averaged similarity coefficient for particular RAPD was related to the presence and type of tandemly repeated motif in a primer: 0.7-0.8 for heterodimers (GC, AG, CA, GT, CT), 0.5-0.6 for homodimers (CC, GG) and 0.6 for no repeat, indicating the sensitivity of diversity range to the type of target sequences. PMID- 17075704 TI - Activity-dependent suppression of spontaneous spike generation in the Retzius neurons of the leech Hirudo medicinalis L. AB - We report on factors affecting the spontaneous firing pattern of the identified serotonin-containing Retzius neurons of the medicinal leech. Increased firing activity induced by intracellular current injection is followed by a 'post stimulus-depression' (PSD) without spiking for up to 23 s. PSD duration depends both on the duration and the amplitude of the injected current and correlates inversely with the spontaneous spiking activity. In contrast to serotonin containing neurons in mammals, serotonin release from the Retzius cells presumably does not mediate the observed spike suppression in a self-inhibitory manner since robust PSD persists after synaptic isolation. Moreover, single additional spikes elicited at specific delays after spontaneously occurring action potentials are sufficient to significantly alter the firing pattern. Since sub-threshold current injections do not affect the ongoing spiking pattern and PSD persists in synaptically isolated preparations our data suggest that PSD reflects an endogenous and 'spike-dependent' mechanism controlling the spiking activity of Retzius cells in a use-dependent way. PMID- 17075705 TI - Thoracic ossification of ligamentum flavum caused by skeletal fluorosis. AB - Thoracic ossification of ligamentum flavum (OLF) caused by skeletal fluorosis is rare. Only six patients had been reported in the English literature. This study reports findings from the first clinical series of this disease. This was a retrospective study of patients with thoracic OLF due to skeletal fluorosis who underwent surgical management at the authors' hospital between 1993 and 2003. Diagnosis of skeletal fluorosis was made based on the epidemic history, clinical symptoms, radiographic findings, and urinalysis. En bloc laminectomy decompression of the involved thoracic levels was performed in all cases. Cervical open door decompression or lumbar laminectomy decompression was performed if relevant stenosis was present. Neurological status was evaluated preoperatively, at the third day postoperatively, and at the end point of follow up using the Japanese Orthopaedic Association (JOA) scoring system of motor function of the lower extremities. A total of 23 cases were enrolled, 16 (69.6%) males and 7 (30.4%) females, age ranging from 42 to 72 years (mean 54.8 years). All patients came from a high-fluoride area, and 22 (95.7%) had dental fluorosis. Medical imaging showed OLF together with ossification of many ligaments and interosseous membranes, including interosseous membranes of the forearm (18/23 patients 78.3%), leg (14/23 patients 60.9%), and ribs (11/23 patients 47.8%). OLF was classified into five types based on MRI findings: localized (4/23 patients 17.4%), continued (12/23 patients 52.2%), skip (3/23 patients 13.0%), combining with anterior pressure (2/23 patients 8.7%), and combining with cervical and/or lumbar stenosis (2/23 patients, 8.7%). Urinalysis showed a markedly high urinary fluoride level in 14 of 23 patients (60.9%). Patients were followed up for an average duration of 4 years, 5 months. Paired t-test showed that the JOA score was slightly but nonsignificantly increased relative to preoperative measurement 3 days after surgery (P = 0.0829) and significantly increased at the end of follow-up (P = 0.0001). In conclusion, Fluorosis can cause ossification of thoracic ligamentum flavum, as well as other ligaments. Comparing with other OLF series, a larger number of spinal segments were involved. The diagnosis of skeletal fluorosis was made by the epidemic history, clinical symptom, imaging study findings, and urinalysis. En bloc laminectomy decompression was an effective method. PMID- 17075706 TI - A perennial ryegrass CBF gene cluster is located in a region predicted by conserved synteny between Poaceae species. AB - CBF/DREB1 proteins are the most important regulators of the cold temperature signaling pathway in many plants. CBF genes are candidates for low-temperature tolerance QTL in wheat and barley. Ten novel putative CBF cDNAs of perennial ryegrass (Lolium perenne L.) have been isolated from cold-treated leaf tissue. Their primary structures contain some conserved motifs, characteristic of the gene class. Phylogenetic analysis revealed that LpCBF genes were attributable to the HvCBF3-, and HvCBF4-subgroups following the previously proposed classification of barley CBF genes. RT-PCR analysis revealed that the expression of LpCBF genes was rapidly induced in response to low temperature and that the expression pattern under the low-temperature conditions for a long period was different between the various LpCBF genes. Five of the ten LpCBF genes were assigned to the genetic linkage map using the p150/112 reference mapping population. LpCBFIb, LpCBFII, LpCBFIIIb and LpCBFIIIc were mapped on LG5 forming a cluster within 2.2 cM, while LpCBFVb was located on LG1. Based on comparative genetic studies, conserved synteny for CBF gene family was observed between the Triticeae cereals and perennial ryegrass. Information on the perennial ryegrass CBF genes at both the molecular and genetic level obtained in this study would be useful for the further study on the role of CBF genes and low-temperature tolerance in grasses. PMID- 17075707 TI - [Medicinal prevention of gastrointestinal tumors: aspirin, Helicobacter and more?]. AB - Despite the huge number of drugs on the market and recent advances in pharmacotherapy, only a few substances are available for the prevention of gastrointestinal tumors--most of which are not approved for this indication or not validated in appropriately designed randomized trials. General recommendations include lifestyle modifications such as avoidance of smoking, only moderate consumption of alcohol, regular physical exercise and a nutrition rich in fresh fruits and vegetables with limited meat. A global eradication therapy for Helicobacter pylori would be desirable to prevent gastric carcinoma, but this does not seem feasible from the socio-economic point of view. Therefore, at least patients at high risk should be screened and this pathogen eradicated, preferentially in their youth. Hepatitis B vaccination of newborns to prevent the development of hepatocellular carcinoma has already been established in Germany; a specific antiviral therapy should be offered to all patients with hepatitis B or C infections, taking into consideration the risks associated with this treatment. The use of non-steroidal anti-inflammatory drugs (NSAIDs) to prevent gastrointestinal malignancies cannot generally be recommended and should be restricted to patients at high risk and to clinical studies. However, the appropriate substance, dose and duration of NSAID therapy are still being debated. PMID- 17075708 TI - [A 52-year-old patient with positive troponin, iron deficiency anemia and known sarcoidosis]. AB - We report on a 52-year-old female patient with recurrent neurosarcoidosis and an "atypical" course of celiac disease, with mild clinic features, positive IgA antibodies and histology. In our patient, the IgA-antibodies led to a false positive troponin I test, and we initially suspected an acute coronary syndrome. With dietary treatment of the celiac disease, the antibodies decreased and the troponin I test became negative. The coincidental occurrence of sarcoidosis and "potential" celiac disease (positive antibodies only) has been reported. The coincidence of an "atypical" celiac disease and sarcoidosis is rare. PMID- 17075709 TI - Mutation, selection, and ancestry in branching models: a variational approach. AB - We consider the evolution of populations under the joint action of mutation and differential reproduction, or selection. The population is modelled as a finite type Markov branching process in continuous time, and the associated genealogical tree is viewed both in the forward and the backward direction of time. The stationary type distribution of the reversed process, the so-called ancestral distribution, turns out as a key for the study of mutation-selection balance. This balance can be expressed in the form of a variational principle that quantifies the respective roles of reproduction and mutation for any possible type distribution. It shows that the mean growth rate of the population results from a competition for a maximal long-term growth rate, as given by the difference between the current mean reproduction rate, and an asymptotic decay rate related to the mutation process; this tradeoff is won by the ancestral distribution. We then focus on the case when the type is determined by a sequence of letters (like nucleotides or matches/mismatches relative to a reference sequence), and we ask how much of the above competition can still be seen by observing only the letter composition (as given by the frequencies of the various letters within the sequence). If mutation and reproduction rates can be approximated in a smooth way, the fitness of letter compositions resulting from the interplay of reproduction and mutation is determined in the limit as the number of sequence sites tends to infinity. Our main application is the quasispecies model of sequence evolution with mutation coupled to reproduction but independent across sites, and a fitness function that is invariant under permutation of sites. In this model, the fitness of letter compositions is worked out explicitly. In certain cases, their competition leads to a phase transition. PMID- 17075710 TI - [Surgical management of peripheral arterial disease. Operative methods and results]. AB - Various operative and interventional methods are available to treat patients with peripheral arterial disease (PAD). The selection of the appropriate therapy should be made after a careful review of the patient's general condition, the morphology of the arterial occlusion, the risk of possible complications, and the likelihood of long-term success for each type of treatment. The different procedures complement one another in their technical possibilities and their risk profile The combination of surgical and interventional methods offers new therapeutic possibilities. The different surgical procedures and their long-term outcome are presented in this publication. PMID- 17075712 TI - [Evidence-based psychotherapy of schizophrenic disorders]. AB - In the past, treatment of schizophrenic disorders was limited to pharmacotherapy. Psychotherapy was not regarded as evidence-based. A number of randomised clinical trials have led to a different perspective during the last two decades. This paper highlights and discusses evidence-based strategies. In particular, cognitive behavioural therapy and family intervention have recently been recommended in evidence-based treatment guidelines. Additionally, psychoeducation and social skills training showed treatment effects. Psychotherapeutic treatment requires an individual case concept. Neuropsychological testing for cognitive deficits should be included in comprehensive assessment in order to avoid overstimulation and optimise rehabilitation. Major treatment goals are to reduce persistent symptoms and relapse rates. PMID- 17075711 TI - [New developments in joint ultrasonography]. AB - Musculoskeletal ultrasonography has become an established imaging technique in rheumatology. The ability of ultrasonography to visualize soft tissue changes provides a possibility for differentiating between exudative and proliferative synovial tissue changes. Superficial cartilage and bone lesions can be detected earlier by ultrasonography than by conventional radiography. The application of Doppler and power Doppler ultrasonography is helpful for the detection of early inflammation. Current studies with ultrasound contrast media demonstrate its benefit in the differentiation of inflammatory processes. Musculoskeletal ultrasonography is helpful in the diagnosis of early arthritis, especially in patients with inconspicuous conventional radiography or suspicious clinical findings. It is a convenient method for follow-up analysis, and therefore has an impact on the monitoring of therapy. It is patient-friendly and is an important tool for the diagnostic work-up of arthritis. PMID- 17075713 TI - [Video-assisted diagnostic thoracoscopy]. AB - Modern radiologic diagnostics show a variety of pathological changes in the mediastinum, pleura, and lung but no evidence on their histogenesis. Transbronchial and transthoracal fine-needle aspiration biopsy usually cannot yield detailed diagnostic results because of its small size. Sufficient and representative material can be obtained by thoracoscopy. Video-assisted thoracoscopy allows safe and fast diagnosis of diffuse lung diseases, pleural diseases including malignant mesothelioma, indeterminate peripheral lung nodule, and mediastinal masses. This gentle diagnostic method can give invaluable information guiding further management of the thoracic injury. Video-assisted thoracoscopy is a safe and effective guiding tool if performed by experienced thoracic surgeons able to convert to thoracotomy. It is to be noted that interpretation of intraoperative findings plays a decisive role in interdisciplinary diagnostics of intrathoracal diseases. PMID- 17075714 TI - Hormone receptor status as a prognostic factor affecting the outcome of brain metastases. PMID- 17075715 TI - [Hereditary photodermatoses]. AB - Hereditary photodermatoses are characterized by an increased photosensitivity caused by an inherited single gene defect. With few exceptions, they manifest in early childhood, reveal heterogeneous clinical symptoms, and are difficult to treat. Although these diseases are rare, it is very important to make an accurate diagnosis on the basis of clinical symptoms, specific diagnostic tests, and direct DNA analysis. We review the spectrum of inherited photodermatoses, including porphyria cutanea tarda, erythropoietic protoporphyria, actinic prurigo, Kindler syndrome, and disorders associated with a defect in DNA repair, including xeroderma pigmentosum, trichothiodystrophy, Cockayne syndrome, and Bloom syndrome. Early diagnosis may prevent complications associated with prolonged unprotected exposure to sunlight and makes it possible to offer genetic counseling and, when indicated, prenatal diagnosis to families at risk for these rare heritable disorders. PMID- 17075716 TI - A novel locus for maternally inherited human gingival fibromatosis at chromosome 11p15. AB - Human isolated gingival fibromatosis is an oral disorder characterized by a slowly progressive benign enlargement of gingival tissues. The most common genetic form, hereditary gingival fibromatosis (HGF), is usually transmitted as an autosomal dominant trait. We report here for the first time a newly identified maternally inherited gingival fibromatosis in two unrelated Chinese families and mapped this disease locus to human chromosome 11p15 with a maximum two point LOD score of 8.70 at D11S4046 (theta = 0) for family 1 and of 6.02 at D11S1318 for family 2. Haplotype analysis placed the critical region in the interval defined by D11S1984 and D11S1338. A cluster of maternally expressed genes is within this critical region. We screened individuals in these two families for mutations for all known maternally expressed genes within this region. None was found either within the coding sequence or at the intron-exon boundary of these genes. Neither did we detect any loss of imprinting in three informative imprinted genes including H19, KCNQ1 downstream neighbor (KCNQ1DN) and cyclin-dependent kinase inhibitor 1C (CDKN1C). However, gene expression profile analysis revealed reduced expression of hemoglobin beta (HBB), hemoglobin delta (HBD), hemoglobin gamma A (HBG1) and hemoglobin gamma G (HBG2) genes at disease locus in HGF patients. This study suggests that genome imprinting might affect the development of HGF. PMID- 17075717 TI - Y-chromosomal insights into the genetic impact of the caste system in India. AB - The caste system has persisted in Indian Hindu society for around 3,500 years. Like the Y chromosome, caste is defined at birth, and males cannot change their caste. In order to investigate the genetic consequences of this system, we have analysed male-lineage variation in a sample of 227 Indian men of known caste, 141 from the Jaunpur district of Uttar Pradesh and 86 from the rest of India. We typed 131 Y-chromosomal binary markers and 16 microsatellites. We find striking evidence for male substructure: in particular, Brahmins and Kshatriyas (but not other castes) from Jaunpur each show low diversity and the predominance of a single distinct cluster of haplotypes. These findings confirm the genetic isolation and drift within the Jaunpur upper castes, which are likely to result from founder effects and social factors. In the other castes, there may be either larger effective population sizes, or less strict isolation, or both. PMID- 17075719 TI - Responses to dipole stimuli of anterior lateral line nerve fibres in goldfish, Carassius auratus, under still and running water conditions. AB - We investigated how fibres in the anterior lateral line nerve of goldfish, Carassius auratus, respond to sinusoidal water motions in a background of still or running water. Two types of fibres were distinguished: type I fibres, which most likely innervate superficial neuromasts, were stimulated by running water (10 cm s(-1)) while type II fibres, which most likely innervate canal neuromasts, were not stimulated by running water. The responses of type I fibres to sinusoidal water motions were masked in running water whereas responses of type II fibres were not masked. These findings are in agreement with previous data obtained from the posterior lateral line nerve of goldfish. Furthermore, we demonstrate here that for type I fibres the degree of response masking increased with increasing flow velocity. Finally, the ratio between responses that were masked in running water (type I) and those that were not masked (type II) increases with increasing flow velocity. Flow fluctuations that were generated by a cylinder in front of the fish did not affect ongoing activity in the flow, nor the dipole-evoked responses. The findings are discussed with respect to particle image velocimetry data of the water motions generated in the experiments. PMID- 17075720 TI - The morphology, physiology and function of suboesophageal neck motor neurons in the honeybee. AB - We report some of the neural and muscular circuitry that allows honeybees to control head movements. We studied neck motor neurons with cell bodies in the suboesophageal ganglion, axons in the first cervical nerve (IK1) and terminals in neck muscles 44 and 51 (muscle classification: Snodgrass in Smithsonian Misc Coll 103:1-120, 1942). We show that muscle 44 actually comprises five separate bundles of muscle fibres (subunits), while muscle 51 is split into two subunits. Eight motor neurons innervate muscles 44 and 51. Two motor neurons have cell bodies in the ventral-median cell body group (one innervates a subunit in muscle 44, the other a subunit in muscle 51). One motor neuron has a ventrally located contralateral cell body (innervating a subunit in muscle 44) and five have laterally located ipsilateral cell bodies. Of the five lateral cells, one innervates a subunit in muscle 51, three selectively innervate subunits in muscle 44 and one co-innervates a subunit in muscle 44 with the contralateral cell. Extracellular recordings revealed three types of visually driven, direction selective cell-types in each IK1 tuned for leftward, rightward and downward motion over the eyes. The spatiotemporal tuning of the units is similar to that of other visual interneurons in the bee brain. PMID- 17075718 TI - Why are mineralocorticoid receptor antagonists cardioprotective? AB - Two clinical trials, the Randomized ALdosterone Evaluation Study (RALES) and the EPlerenone HEart failure and SUrvival Study (EPHESUS), have recently shown that mineralocorticoid receptor (MR) antagonists reduce mortality in patients with heart failure on top of ACE inhibition. This effect could not be attributed solely to blockade of the renal MR-mediated effects on blood pressure, and it has therefore been proposed that aldosterone, the endogenous MR agonist, also acts extrarenally, in particular in the heart. Indeed, MR are present in cardiac tissue, and possibly aldosterone synthesis occurs in the heart. This review critically addresses the following questions: (1) is aldosterone synthesized at cardiac tissue sites, (2) what agonist stimulates cardiac MR normally, and (3) what effects are mediated by aldosterone/MR in the heart that could explain the beneficial effects of MR blockade in heart failure? Conclusions are that most, if not all, of cardiac aldosterone originates in the circulation (i.e., is of adrenal origin), and that glucocorticoids, in addition to aldosterone, may serve as the endogenous agonist of cardiac MR. MR-mediated effects in the heart include effects on endothelial function, cardiac fibrosis and hypertrophy, oxidative stress, cardiac inotropy, coronary flow, and arrhythmias. Some of these effects occur via or in synergy with angiotensin II, and involve a non-MR-mediated mechanism. This raises the possibility that aldosterone synthase inhibitors might exert beneficial effects on top of MR blockade. PMID- 17075722 TI - Exploration of the emergence of the Victoria lineage of influenza B virus. AB - The Victoria lineage represented by B/Victoria/2/87 is one of the two major distinctive haemagglutinin (HA) lineages of influenza B virus, and its recent re emergence has aroused great concerns. However, it remains unknown when, where, and how this HA lineage emerged in the world. In this study, the HA1 domain of the HA gene of fourteen influenza B viruses isolated in China in 1972-1984 was sequenced. The sequences were phylogenetically analyzed with the HA1 sequences of 41 other important influenza B isolates. The results unveiled some earlier footprints of the Victoria lineage in China, and the epidemic history of the Victoria lineage could be traced back from the year 1985 to 1975. Moreover, phylogenetic analysis, the history of China, and the epidemiology of influenza B virus indicated that the Victoria lineage possibly emerged in China in the 1970s through gradual evolution from a minor lineage. PMID- 17075723 TI - Male-produced aggregation pheromone compounds from the eggplant flea beetle (Epitrix fuscula): identification, synthesis, and field biossays. AB - Volatiles from the eggplant flea beetle, Epitrix fuscula Crotch (Coleoptera: Chrysomelidae), feeding on host foliage, were investigated. Six male-specific compounds were detected and were identified through the use of mass spectrometry, nuclear magnetic resonance (NMR) spectrometry, chiral and achiral gas chromatography, high-performance liquid chromatography, electrophysiology (gas chromatography-electroantennography, GC-EAD), and microchemical tests. The two most abundant of the six compounds were (2E,4E,6Z)-2,4,6-nonatrienal (1) and (2E,4E,6E)-2,4,6-nonatrienal (2). The other four compounds, present in minor amounts, were identified as himachalene sesquiterpenes; two of these, 3 and 4, were hydrocarbons and two, 5 and 6, were alcohols. All four sesquiterpenes were previously encountered from male flea beetles of Aphthona spp. and Phyllotreta cruciferae. Synthetic 1 and 2 matched the natural products by GC retention times, mass spectra, and NMR spectra. Sesquiterpenes 3-6 similarly matched synthetic standards and natural samples from the previously studied species in all ways, including chirality. Both natural and synthetic 1 and 2 gave positive GC-EAD responses, as did sesquiterpenes 3, 5, and 6. Field trials were conducted with a mixture of 1 and 2, and the baited traps were significantly more attractive than control traps to both male and female E. fuscula. The E. fuscula pheromone has potential for monitoring or controlling these pests in eggplants. PMID- 17075721 TI - Gender dependent APP processing in a transgenic mouse model of Alzheimer's disease. AB - Epidemiological studies have reported a higher prevalence and incidence of Alzheimer's disease (AD) in women. The biochemical basis for this gender disparate susceptibility is unknown. A gender effect on AD-typical plaque pathology has been shown in APP transgenic mouse models of AD. Female mice elicit higher plaque load than male mice. In an effort to analyze gender-dependent APP processing during postnatal development, we examined APP transgenic mice at time points prior to plaque deposition. At 14 weeks of age there was a significant elevation of C99 and Abeta in female mice compared to males. Furthermore we observed a slight decrease of BACE-activity in male mice as well as higher cerebral manganese levels in females. Although the decline in estrogen levels due to menopause in female patients is still discussed to be a risk factor for AD our results implicates that additional factors like modified BACE-activity or metal levels may contribute to the higher prevalence and incidence of AD in females. PMID- 17075724 TI - Sex pheromone of the pine sawfly, Gilpinia pallida: chemical identification, synthesis, and biological activity. AB - We present the identification of the sex pheromone in the pine sawfly, Gilpinia pallida, including analysis of the female pheromone content, male antennal response and attraction in the field, and synthesis of the most active pheromone component. Several 3,7-dimethyl-2-alkanols were identified from female whole-body extracts, including some compounds with a 2R configuration. This is the first observation of such compounds in a pine sawfly species. Antennae of male G. pallida responded strongly in electroantennograph (EAG) recordings to the (2S,3R,7R)-isomers of the propionates of 3,7-dimethyl-2-tridecanol, 3,7-dimethyl 2-tetradecanol, and 3,7-dimethyl-2-pentadecanol, as well as to the acetates of the tri- and pentadecanols (the acetate of the tetradecanol was not tested). The propionate of (2S,3R,7R)-3,7-dimethyl-2-tetradecanol caught more males in the field than the corresponding isomer of tri- or pentadecanol. We suggest that the (2S,3R,7R)-isomer of 3,7-dimethyl-2-tetradecanol is likely the main sex pheromone precursor in G. pallida, with a subsidiary role for the (2S,3R,7R)-isomer of the tridecanol. Preparation of highly pure (2R,3R,7R)- and (2S,3R,7R)-stereoisomers of 3,7-dimethyl-2-tetradecanol, including the biological active esters, was performed via chemoenzymatic methods and is described in detail. PMID- 17075725 TI - Increased anti-oxidative potency of garlic by spontaneous short-term fermentation. AB - Fundamental anti-oxidative properties of 80% ethanol extract from garlic fermented for the relatively short period of time (40 days at 60-70 degrees C, 85 95% relative humidity) were examined. Superoxide dismutase (SOD)-like activity, scavenging activity against hydrogen peroxide and the polyphenol content of the garlic extract were increased 13-folds, more than 10-folds, and 7-folds, respectively, as compared with those of the control garlic extract. The results indicate that relatively short-term spontaneous fermentation potentiates anti oxidative properties of garlic in fresh form, which is, at least in part, attributable to the increased level of polyphenols. Since superoxide is the primary upstream radical of the chain reaction with reactive oxygen species and hydrogen peroxide is generated from the scavenging reaction by SOD, the fermented garlic is suggested to possess desirable anti-oxidative properties. PMID- 17075726 TI - Prevalence and correlates of hepatitis C virus infection among street-recruited injection drug users in San Juan, Puerto Rico. AB - Throughout the world, injection drug users (IDUs) are the group at highest risk for hepatitis C virus (HCV) infection. IDUs residing in the island of Puerto Rico and Puerto Rican IDUs residing in the U.S. mainland have been shown to be at very high risk of infection with HIV. However, the extent to which HCV infection has spread among IDUs in Puerto Rico is not yet known. The aims of this study were to estimate seroprevalence of HCV and to identify the correlates associated with HCV transmission. The sample was drawn through street outreach strategies and was comprised of 400 injection drug users not in treatment, living in the San Juan metropolitan area. HCV and HIV infection were detected by enzyme-linked immunosorbent assay and the results were confirmed by Western blot. Information on sociodemographics, drug use patterns, and risk behaviors was obtained through structured interviews. Bivariate analyses and multivariate logistic regression were used to assess covariates of infection with HCV. The prevalence of HCV infection was 89%. After controlling for sociodemographic characteristics, HCV infection was positively associated with increasing years of injection, injecting in a shooting gallery, tattooing in prison, and self-reported STD infection. Notably, IDUs who had initiated drug injection within the year prior to the study interview had an HCV infection rate of 57%. This study indicates that more aggressive educational programs are urgently needed to reduce the spread of HCV infection among IDUs in Puerto Rico. PMID- 17075727 TI - An analysis of respondent driven sampling with Injection Drug Users (IDU) in Albania and the Russian Federation. AB - Injection drug users in Tirana, Albania and St. Petersburg, Russia were recruited into a study assessing HIV-related behaviors and HIV serostatus using Respondent Driven Sampling (RDS), a peer-driven recruitment sampling strategy that results in a probability sample. (Salganik M, Heckathorn DD. Sampling and estimation in hidden populations using respondent-driven sampling. Sociol Method. 2004;34:193 239). This paper presents a comparison of RDS implementation, findings on network and recruitment characteristics, and lessons learned. Initiated with 13 to 15 seeds, approximately 200 IDUs were recruited within 8 weeks. Information resulting from RDS indicates that social network patterns from the two studies differ greatly. Female IDUs in Tirana had smaller network sizes than male IDUs, unlike in St. Petersburg where female IDUs had larger network sizes than male IDUs. Recruitment patterns in each country also differed by demographic categories. Recruitment analyses indicate that IDUs form socially distinct groups by sex in Tirana, whereas there was a greater degree of gender mixing patterns in St. Petersburg. RDS proved to be an effective means of surveying these hard-to reach populations. PMID- 17075731 TI - Follow-up of sex reassignment surgery in transsexuals: a Brazilian cohort. AB - This study examined the impact of sex reassignment surgery on the satisfaction with sexual experience, partnerships, and relationship with family members in a cohort of Brazilian transsexual patients. A group of 19 patients who received sex reassignment between 2000 and 2004 (18 male-to-female, 1 female-to-male) after a two-year evaluation by a multidisciplinary team, and who agreed to participate in the study, completed a written questionnaire. Mean age at entry into the program was 31.21+/-8.57 years and mean schooling was 9.2+/-1.4 years. None of the patients reported regret for having undergone the surgery. Sexual experience was considered to have improved by 83.3% of the patients, and became more frequent for 64.7% of the patients. For 83.3% of the patients, sex was considered to be pleasurable with the neovagina/neopenis. In addition, 64.7% reported that initiating and maintaining a relationship had become easier. The number of patients with a partner increased from 52.6% to 73.7%. Family relationships improved in 26.3% of the cases, whereas 73.7% of the patients did not report a difference. None of the patients reported worse relationships with family members after sex reassignment. In conclusion, the overall impact of sex reassignment surgery on this cohort of patients was positive. PMID- 17075732 TI - Seasonal viral loop dynamics in two large ultraoligotrophic Antarctic freshwater lakes. AB - The effect of viruses on the microbial loop, with particular emphasis on bacteria, was investigated over an annual cycle in 2003-2004 in Lake Druzhby and Crooked Lake, two large ultraoligotrophic freshwater lakes in the Vestfold Hills, Eastern Antarctica. Viral abundance ranged from 0.16 to 1.56 x 10(9) particles L 1 and bacterial abundances ranged from 0.10 to 0.24 x 10(9) cells L-1, with the lowest bacterial abundances noted in the winter months. Virus-to-bacteria ratios (VBR) were consistently low in both lakes throughout the season, ranging from 1.2 to 8.4. lysogenic bacteria, determined by induction with mitomycin C, were detected on three sampling occasions out of 10 in both lakes. In Lake Druzhby and Crooked Lake, lysogenic bacteria made up between 18% and 73% of the total bacteria population during the lysogenic events. Bacterial production ranged from 8.2 to 304.9 x 10(6) cells L-1 day-1 and lytic viral production ranged from 47.5 to 718.4 x 10(6) viruslike particles L-1 day-1. When only considering primary production, heterotrophic nanoflagellate (HNF) grazing and viral lysis as the major contributors to the DOC pool (i.e., autochthonous sources), we estimated a high contribution from viruses during the winter months when >60% of the carbon supplied to the DOC pool originated from viral lysis. In contrast, during the summer <20% originated from viral lysis. Our study shows that viral process in ultraoligotrophic Antarctic lakes may be of quantitative significance with respect to carbon flow especially during the dark winter period. PMID- 17075733 TI - Effect of microbial species richness on community stability and community function in a model plant-based wastewater processing system. AB - Microorganisms will be an integral part of biologically based waste processing systems used for water purification or nutrient recycling on long-term space missions planned by the National Aeronautics and Space Administration. In this study, the function and stability of microbial inocula of different diversities were evaluated after inoculation into plant-based waste processing systems. The microbial inocula were from a constructed community of plant rhizosphere associated bacteria and a complexity gradient of communities derived from industrial wastewater treatment plant-activated sludge. Community stability and community function were defined as the ability of the community to resist invasion by a competitor (Pseudomonas fluorescens 5RL) and the ability to degrade surfactant, respectively. Carbon source utilization was evaluated by measuring surfactant degradation and through Biolog and BD oxygen biosensor community level physiological profiling. Community profiles were obtained from a 16S-23S rDNA intergenic spacer region array. A wastewater treatment plant-derived community with the greatest species richness was the least susceptible to invasion and was able to degrade surfactant to a greater extent than the other complexity gradient communities. All communities resisted invasion by a competitor to a greater extent than the plant rhizosphere isolate constructed community. However, the constructed community degraded surfactant to a greater extent than any of the other communities and utilized the same number of carbon sources as many of the other communities. These results demonstrate that community function (carbon source utilization) and community stability (resistance to invasion) are a function of the structural composition of the community irrespective of species richness or functional richness. PMID- 17075734 TI - An indigenous drought-tolerant strain of Glomus intraradices associated with a native bacterium improves water transport and root development in Retama sphaerocarpa. AB - The effects of interactions between Bacillus thuringiensis, a drought-adapted bacterium, and two isolates of Glomus intraradices, an arbuscular mycorrhizal (AM) fungus, on Retama sphaerocarpa, a drought-adapted legume, were investigated. The fungal isolates were an indigenous drought-tolerant and a nonindigenous drought-sensitive isolate. Shoot length and root growth, symbiotic parameters, water transport (in terms of percent relative plant water uptake), and volumetric soil moisture and soil enzymatic activities in response to microbial inoculations were evaluated. Retama plants colonized by G. intraradices plus Bacillus possessed similar shoot length after 30 days from sowing compared with noninoculated Retama plants after 150 days. Inoculation with drought-adapted bacterium increased root growth by 201%, but maximum root development was obtained by co-inoculation of B. thuringiensis and the indigenous G. intraradices. Nodules were formed only in plants colonized by autochthonous AM fungi. Relative water uptake was higher in inoculated than in noninoculated Retama plants, and these inoculants depleted soil water content concomitantly. G. intraradices-colonized Retama reached similar shoot length irrespective of the fungal origin, but there were strong differences in relative water uptake by plants colonized by each one of the fungi. Indigenous G. intraradices-colonized roots (evaluated as functional alkaline phosphatase staining) showed the highest intensity and arbuscule richness when associated with B. thuringiensis. The interactive microbial effects on Retama plants were more relevant when indigenous microorganisms were involved. Co-inoculation of autochthonous microorganisms reduced by 42% the water required to produce 1 mg of shoot biomass. This is the first evidence of the effectiveness of rhizosphere bacterium, singly or associated with AM fungus, in increasing plant water uptake, which represents a positive microbial effect on plants grown under drought environments. PMID- 17075735 TI - The logic model as a framework for community program evaluations: the food literacy partners program. PMID- 17075736 TI - More on scientific assembly drug company support. PMID- 17075738 TI - Family medicine residents and home visits. PMID- 17075737 TI - Support for commentary on AAFP scientific assembly. PMID- 17075739 TI - Teaching preventive medicine in an office-based visit. PMID- 17075740 TI - Teaching medical students to prioritize preventive services. AB - BACKGROUND: Clinicians cannot provide all recommended preventive services in a single office visit and must learn to prioritize. This skill is not overtly addressed in medical school. METHODS: We designed a workshop to teach third-year medical students to prioritize preventive services during an office visit. In a prospective controlled trial, we compared performance on a standardized patient case. RESULTS: Students performed well, but there was no significant difference between intervention and control groups' mean scores on the standardized patient encounter. CONCLUSIONS: Our brief intervention failed to increase students' scores on a standardized patient case requiring preventive services prioritization. PMID- 17075741 TI - An "uneventful" pregnancy. PMID- 17075742 TI - Use of a writing elective to teach cultural competency and professionalism. PMID- 17075743 TI - Will rural family medicine residency training survive? AB - BACKGROUND AND OBJECTIVES: Rural family medicine residencies may be more threatened by declining interest in family medicine than their urban counterparts. This study examines the recent performance of rural residencies in the National Resident Matching Program as an indicator of their viability. METHODS: We surveyed all 30 family medicine residencies located in rural areas during the summer of 2004 and a geographically matched sample of 31 urban residencies. We gathered information about the matching process for 2002, 2003, and 2004. The response rate was 70.5%. RESULTS: Rural programs offer about one third fewer first-year (postgraduate year 1 [PGY-1]) positions than their urban counterparts. Rural programs had lower Match rates (60.1%) than urban programs (72.5%) in 2004 but no meaningful differences in the proportion of international medical graduates (IMGs) or osteopathic physicians (DOs) who ultimately accepted positions. The 44.2% of residencies that predicted they would be thriving 2 years in the future filled an average of 81.3% of their slots on Match Day; there were no rural/urban differences. Programs with less-optimistic appraisals of their future had much lower Match rates. Two factors were associated with lower Match rates when other variables were taken into account: the proportion of IMGs in the 2 previous entering years and a stated rural mission. CONCLUSIONS: Rural programs appear to be slightly less stable than their urban counterparts, but the differences are minor. The viability of rural family medicine residency programs is probably affected more by the overall attractiveness of family medicine as a discipline rather than the rural or urban location of the residency. PMID- 17075744 TI - Deaf women's experiences and satisfaction with prenatal care: a comparative study. AB - BACKGROUND AND OBJECTIVES: The quality of communication between physician and patient is a major contributor to patient satisfaction and treatment adherence. Deaf patients who use American Sign Language experience significant communication barriers in most medical settings. This study investigated factors impacting deaf patients' satisfaction with prenatal care and prenatal care disparities between deaf and hearing women. METHODS: Questionnaires modified from Omar and Schiffman's prenatal satisfaction measure were administered to 23 deaf and 32 hearing women. RESULTS: Deaf women were less satisfied than hearing women with physician communication and less satisfied with overall care. Deaf women's expectations about provision of interpreter services being met or exceeded was significantly associated with overall satisfaction. Hearing women had more prenatal care appointments and reported receiving more information from their doctors. CONCLUSIONS: Maximizing communication effectiveness with deaf patients results in better prenatal care and improved patient satisfaction. Good communication includes conveying concern and making efforts to ensure that whatever communication methods used are effective. PMID- 17075745 TI - Health literacy is a predictor of HIV/AIDS knowledge. AB - BACKGROUND AND OBJECTIVES: This study's objective was to evaluate the association between health literacy and human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) knowledge among patients seen at an inner city, public hospital urgent care center (UCC). METHODS: We used a prospective survey of patients offered an HIV test by their providers during a UCC visit. We measured patients' health literacy level using the Rapid Estimate of Adult Literacy in Medicine (REALM) scale and assessed their HIV/AIDS knowledge using a 22-item questionnaire. RESULTS: A total of 372 patients were enrolled. Among participants in this relatively young sample (55% were under the age of 40), 92 (25%) had a REALM score at or below a sixth-grade level, and 122 (33%) did not have a high school diploma. Patients' mean HIV/AIDS knowledge scores differed significantly between patients with inadequate health literacy and those with marginal or adequate health literacy. In multivariate analyses, patients' REALM scores were positively associated with patients' HIV/AIDS knowledge even after adjusting for income, education, and risk perception. CONCLUSIONS: These findings demonstrate that HIV/AIDS knowledge is strongly associated with patients' health literacy in this inner-city population. These findings reiterate the need to target HIV prevention strategies toward populations with inadequate health literacy levels and to dispel misconceptions regarding HIV/AIDS that directly influence risk-taking behaviors and health care utilization. PMID- 17075746 TI - Survey of osteoporosis preventive care in community family medicine settings. AB - OBJECTIVE: This study's objective was to document and describe osteoporosis preventive care for women age 45 years and older in community family medicine practices. METHODS: We conducted a crosssectional mailed survey of 400 women age 45 years and older enrolled in a community-based family medicine research network. Participants responded to 42 items regarding osteoporosis screening and prevention during primary care visits. RESULTS: A total of 275 women returned the survey (response rate 71.4%). Of the respondents, 162 (58.9% of the sample) were ages 45 to 64, and 113 (41.1%) were age 65 and older. Rates of counseling on calcium intake, exercise, falls, and bone density testing were similar in the two age groups. Half of women age 65 and older and 43.8% of women under 65 had received bone density testing. Ninety-two percent of the respondents rated a discussion of osteoporosis and fracture prevention with their primary care provider as "very," "moderately," or "somewhat" important, but only 44% actually had such a discussion. CONCLUSIONS: Most women age 45 and older considered osteoporosis preventive care to be important. However, fewer than half discussed this topic with their primary care provider, and only half of women age 65 and older had undergone bone density screening. PMID- 17075747 TI - Teaching the outstanding medical learner. AB - Medical students and residents are typically prescribed a standardized curriculum. Learners who have difficultly meeting requirements of this curriculum often require extra attention and an individualized program of remediation. Outstanding medical learners, on the other hand, are rarely given an individualized learning program, even if they can easily master the standardized curriculum. These learners are likely to belong to a group that educators call "gifted adults." No literature on this group of medical learners currently exists, and no research has been conducted on them. This article is designed to begin a dialog on the best approach to educating outstanding medical learners using the framework of the gifted adult. Gifted adults possess a unique set of characteristics and have needs that set them apart from the majority of medical learners. There are also well-described pitfalls that these learners encounter. Gifted adults can best exercise their talents and maximize their potential when they receive an individualized educational approach that focuses on providing intellectual challenges with permission to explore interest areas. Research, publishing, teaching opportunities, and leadership roles are some examples of ways to challenge these learners. They should be strongly encouraged to extensively pursue individual interests. Effort should be made to ensure that these learners have contact with peers and mentors of equal intellectual abilities. Gifted adults who are not intellectually stimulated on a routine basis and are not able to interact with other talented individuals may get bored and frustrated with their education. Medical educators who understand the characteristics and drives of gifted adults will be better able to ensure that these learners are regularly challenged, with needed assistance to avoid common pitfalls. PMID- 17075748 TI - Developing an educational reminder system for a handheld encounter log. AB - BACKGROUND AND OBJECTIVES: Medical students increasingly log patient encounters on handheld computers. Detailed patient records captured at the bedside would allow these log programs to complement medical education with automated reminders. We evaluated progress toward this goal. METHODS: A series of electronic student encounter logs (ESEL) were designed to quickly record common diagnoses in a family medicine clerkship. Common diagnoses were organized in a shallow, broad-tree structure that locates many concepts with one to four taps. Third-year clerks' ESEL records of patient problems were evaluated longitudinally and across two similar clerkships. Grading criteria were introduced to encourage attention to target problems, especially infrequently reported psychosocial issues. A pilot reminder system in ESEL-4, the latest version of the computer application program, included 17 high-specificity reminders. Students' viewing of reminders was monitored. RESULTS: Students recorded complex patients in ESEL-4 and entered 80% of patients during office hours. ESEL grading generated anxiety but was associated with significantly increased and plausible documentation of tobacco abuse, depression, and alcohol abuse. Students viewed titles of about 25% of generated reminders but reviewed less than 1% of reminders in detail. CONCLUSIONS: Students using ESEL-4 document plausibly complex patients, usually during office hours. Grading probably improves completeness and accuracy. Diagnosis-driven educational reminder systems are possible but do not yet capture students' attention. PMID- 17075749 TI - Prisoners' dilemma: the importance of negative results. PMID- 17075756 TI - [The tale of intraocular cefuroxime]. PMID- 17075757 TI - [About referees, anonymous, cowardice and bad education]. PMID- 17075758 TI - [Laser ray tracing to measure optical aberrations in eyes with contact lenses]. AB - PURPOSE: To evaluate the impact of disposable soft contact lenses upon visual performance by comparing the optical aberrations of myopic eyes with and without the use of contact lenses. METHOD: The optical aberrations of a set of 18 eyes were measured by a laser ray tracing system, an objective measurement method, enabling comparison of the findings of the naked eye, the eye plus a test lens, and the eye plus a disposable contact lens. RESULTS: There was a large variability among subjects, but the general tendency was that test lenses did not modify aberrations, whereas contact lenses tended to increase the ocular aberrations for large pupils by an average of 40%. There was a lower increase for smaller pupils. CONCLUSION: This study predicts there will be no significant adverse effects on the quality of vision from the use of disposable contact lenses worn under daylight conditions, but a small decline in night vision. PMID- 17075759 TI - [Outcomes of macular hole surgery using three different surgical techniques]. AB - PURPOSE: To compare the effectiveness, efficiency and complications of three different surgical techniques employed for the treatment of macular holes between 1998 -2004 in our health care centre. METHODS: A retrospective, comparative, and non-randomized study of 131 eyes with macular holes, stage III and IV, treated with three different surgical techniques was performed. Vitrectomy was performed in 25 eyes (group 1). Vitrectomy was associated with internal limiting membrane peeling in 19 eyes (group 2), and in the remaining 87 eyes, the peeling of the retinal internal limiting membrane was assisted with indocyanine green (ICG) (group 3). The concentrations of ICG used were 5%, 2.5% and 0.5%. We compared the anatomic and functional results, and the complications which occurred. RESULTS: The overall anatomic success rate was 88.4% (114 eyes), and it was higher in group 3 (90.6%). A visual acuity improvement was achieved in 63.6% of eyes; however there were no significant differences between the groups studied. The most important complications included cataracts (51.9%), increase in intra-ocular pressure (37.2%), retinal detachment (7.8%) and retinal pigment epithelial changes (7%). These pigment epithelial changes were associated with the use of ICG-assisted peeling (p<0.05), but there was no correlation between the changes and the concentration of ICG employed. The hole size had an inverse correlation with initial and final visual acuity. An epi-retinal membrane was associated with better visual acuities and higher anatomic success. CONCLUSIONS: In our experience, the use of ICG-assisted internal limiting membrane peeling in macular hole surgery improves the anatomic success, but may lead to retinal pigment epithelial changes. PMID- 17075760 TI - [Filtering bleb after surgical cyclodialysis]. AB - CASE REPORT: A 35-year-old man with degenerative myopia suffered blunt ocular trauma due to being hit in the face by a ball. The injury caused four cyclodialysis clefts in his left eye which resulted in hypotony and chorioretinal folds. DISCUSSION: The persisting hypotony was unsuccessfully treated with topical therapy and with laser photocoagulation. Therefore, a surgical treatment with direct cyclopexy was performed. In one of the cyclodialysis clefts, incarcerated vitreous was found, and this was considered to be the reason for its incomplete closure. In the postoperative period complete closure of the cyclodialysis cleft was observed; however an unexpected filtering bleb appeared, which could have affected good control of intraocular pressure. PMID- 17075761 TI - [Bilateral central serous chorioretinopathy with subretinal exudation]. AB - CLINICAL CASE: A 37-year-old man was referred because of a bilateral loss of visual acuity and metamorphopsia. On examination there was yellowish exudation bilaterally associated with serous retinal detachments, corresponding to hyperfluorescence spots in the early stage of fluorescein angiography. Photocoagulation of the leakage spots was performed in both eyes. Six months later he had recovered his visual acuity and the lesions had remitted. DISCUSSION: The clinical findings, ancillary tests and subsequent clinical course allowed a diagnosis of Best's disease to be ruled out. The presence of sub retinal yellowish exudation in the posterior pole does not necessarily rule out the diagnosis of central serous chorioretinopathy. PMID- 17075762 TI - [Recurrence of peripheral primitive neuroectodermal tumor of the orbit with systemic metastases]. AB - CASE REPORT: A six-year-old boy presented with proptosis of the right eye. Imaging studies detected a mass in the medial wall of the right orbit. This mass was biopsied revealing a histopathologic diagnosis of primitive neuroectodermal tumor, so chemotherapy treatment was given. After seven years in remission he presented with a recurrence of the orbital tumor and was found to also have systemic metastases. Treatment with chemotherapy, radiotherapy and orbital exenteration was unsuccessful. DISCUSSION: The orbital occurrence of these tumors is extremely rare. Differentiation from other small round cell tumors requires immunohistochemical and ultrastructural techniques. PMID- 17075763 TI - [Treatment of two cases of diffuse retinal pigment epitheliopathy with photodynamic therapy]. AB - CASE REPORT: We present two patients with Diffuse Retinal Pigment Epitheliopathy (DRPE) treated with verteporfin photodynamic therapy (PDT). The first patient was treated with PDT because there was foveal involvement. The second patient received PDT because he had multiple recurrences of DRPE with poor resolution following previous argon-laser photocoagulation. Resolution of the lesions occurred in both cases with improvement in visual acuity. No recurrences have been documented at eleven months of follow-up. DISCUSSION: The outcomes in our patients were satisfactory. We consider PDT is a safe and effective option, but further randomized studies are necessary to confirm this. PMID- 17075764 TI - [Bilateral recurrent autoimmune optical neuropathy in childhood]. AB - CLINICAL CASE: A ten year-old girl, after a Yersinia gastroenteritis, developed an optic neuritis in the left eye. She was not treated and resulted ultimately in optic atrophy on the affected side. Six months later a similar episode occurred in the contralateral eye. On this occasion corticosteroid therapy was given. During this therapy the neuritis diminished; however the patient had three relapses, so it was decided to give her immunosuppressive treatment with azathioprine and continue this indefinitely. DISCUSSION: After considering the differential diagnoses of bilateral recurrent optical neuritis in childhood, we concluded that it was most likely to have an autoimmune basis. After considering the benefits and risks of the long-term treatment, we believe immunosuppressive therapy is most useful in controlling the disease allowing corticosteroid therapy to be reduced. PMID- 17075765 TI - [Atypical frontal sinus mucocele. A case report]. AB - CLINICAL CASE: We report a case of a 46-year-old woman who developed a tender, painful mass in the left superior eyelid over a period of about 6 months. This was a frontal mucocele with atypical clinical and histopathologic features. DISCUSSION: Most mucoceles arise from the frontal or ethmoidal sinuses. Frontal mucoceles usually cause outward and downward displacement of the globe, and are often associated with fullness in the supero-nasal and medial canthal region and a palpable mass. PMID- 17075766 TI - [Sub-tenon steroid treatment in choroidal neovascularization caused by serpiginous choroiditis. A case report]. AB - CASE REPORT: We present a report of a patient suffering from serpiginous choroiditis complicated by choroidal neovascularization (CNV). Sub-tenon steroid treatment, added to the usual immunosuppressive therapy, resolved the CNV, improving visual acuity in one eye. DISCUSSION: Choroidal neovascularization is a sight-threatening complication of posterior uveitis. Our case suggests that sub tenon steroid treatment, added to systemic immunosuppressive therapy, may accelerate the CNV resolution in posterior uveitis. PMID- 17075767 TI - [Eye anatomy in the 18th Century]. PMID- 17075768 TI - [World Day for White Rods and the al Noor Association in the Oman Sultanate]. PMID- 17075769 TI - Transient local overexpression of human vascular endothelial growth factor (VEGF) in mouse feto-maternal interface during mid-term pregnancy lowers systemic maternal blood pressure. AB - The effect on maternal circulation of transient human vascular endothelial growth factor (VEGF) (165) cDNA transfection into the mouse feto-maternal interface at day 14.5 post coitus (p.c.) using a hemagglutinating virus of Japan-envelope (HVJ E) vector system is reported. On day 15.5 p.c., Western blotting clearly showed overexpression of 18 kD VEGF protein in the uterus. After VEGF transfection, the blood pressure was significantly lowered for 48 hours. On day 17.5 p.c., the blood pressure returned to the control level. Proteinuria was not observed after VEGF transfection. No preterm birth was observed during the course of pregnancy after the transfection procedure. After 24 hours of transfection, human VEGF was not detectable and the mouse VEGF level was similar to that in peripheral blood. However, the soluble fms-like tyrosine kinase (Flt)-1 concentration was significantly lower in VEGF-transfected mice. These results suggest that extraamniotic VEGF overexpression lowered the systemic blood pressure without altering the VEGF concentration in the peripheral blood. Local overexpression of VEGF may become a novel treatment for pregnancy-related disorders such as hypertension complicated-pregnancy and preeclampsia. PMID- 17075770 TI - Evidence for impaired gluconeogenesis in very long-chain acyl-CoA dehydrogenase deficient mice. AB - Hypoketotic hypoglycaemia is a characteristic feature of fatty acid oxidation (FAO) defects. Although the underlying pathogenic mechanism is unknown, one hypothesis points to an impairment in gluconeogenesis. To study hepatic glucose production in FAO defects, we used the knockout mouse model of very long-chain acyl-CoA dehydrogenase (VLCAD) deficiency presenting with stress-induced hypoglycaemia. We analysed metabolites of hepatic glucose production under non stressed conditions and after stress in comparison to wildtype controls. Analysis included glycogen, glucose-6-phosphate (G6P), fructose-6-phosphate (F6P), glycerol-3-phosphate (G3P) and dihydroxyacetone-phosphate (DHAP). We also measured the activity of the key enzyme glucose-6-phosphatase. Blood and liver glucose were found to be low after stress, and liver glycogen was depleted. In addition, hepatic G6P and F6P were significantly reduced, especially during hypoglycaemia. Importantly, the activity of the enzyme converting G6P into glucose was not impaired. These data indicate a reduced rate of gluconeogenesis. The levels of DHAP and G3P were significantly lower suggesting decreased availability of glucose precursors from glycerol. This study gives biochemical evidence of impaired gluconeogenesis as one of the causes for hypoglycaemia observed in VLCAD deficiency. Whether this is due to lack of a substrate, inhibitory effects on other gluconeogenic enzymes or impaired transcription of gluconeogenic enzymes needs to be resolved in the future. PMID- 17075771 TI - Subcutaneous fat modulates insulin sensitivity in mice by regulating TNF-alpha expression in visceral fat. AB - The distribution of fat in obese persons is related to the risk of developing various metabolic disorders, such as glucose intolerance, dyslipidemia and hypertension, and the combination of these conditions is known as the metabolic syndrome. The aim of this study was to investigate the role of subcutaneous fat in regulating insulin resistance and its influence on TNF-alpha expression in visceral fat, by using mice that were subjected to subcutaneous lipectomy with or without subsequent fat transplantation. After partial subcutaneous lipectomy, mice showed significantly greater accumulation of visceral fat compared with sham operated control mice. Lipectomy led to higher plasma insulin and lower plasma glucose levels after loading with glucose and insulin, respectively, compared with the levels in control mice. Insulin-induced phosphorylation of IRS-1 was decreased in the skeletal muscles of lipectomized mice. Subcutaneous transplantation of fat pads into lipectomized mice reversed the above-mentioned changes indicating insulin resistance in these animals. The fat storage area of adipocytes and TNF- alpha expression by adipocytes in visceral fat were significantly higher in the lipectomized mice than in controls, while subcutaneous transplantation of fat reduced both the fat storage area and TNF alpha expression. The insulin resistance of lipectomized mice was also ameliorated by systemic neutralization of TNF-alpha activity using a specific antibody. These findings obtained in mice subjected to subcutaneous lipectomy with/without subsequent fat transplantation indicate that subcutaneous fat regulates systemic insulin sensitivity, possibly through altering fat storage and the expression of TNF-alpha by adipocytes in visceral fat. The balance between accumulation of subcutaneous fat and visceral fat may be important with respect to the occurrence of systemic insulin resistance in the metabolic syndrome. PMID- 17075772 TI - Ultrastructural effects of low dosage endocrine disrupter chemicals on neural cells of the chicken embryo model. AB - Previous research suggests that endocrine disrupters (EDCs) like nonylphenol cause apoptosis (both via the intrinsic and extrinsic pathway) and that ROS generation and Ca (2+) play a fundamental role in the process. We have investigated morphological changes induced by 17beta-estradiol, nonylphenol, 17alpha-ethynylestradiol and diethylstilbestrol on the IN OVO neural chick embryo model by using transmission and scanning electron microscopy (TEM and SEM). We found that estrogenic substances such as nonylphenol, diethylstilbestrol (DES) and 17alpha-ethynylestradiol, as well as 17beta-estradiol cause ultrastructural changes to developing neurons, resulting in damage to the plasma, mitochondrial as well as nuclear membranes. Furthermore, both apoptotic blebbing and necrotic (or oncotic) budding was seen in TEM and SEM micrographs. SEM shows that nonylphenol-exposed neurons have irregular cell surfaces with pseudopodia, cell shrinkage and breakages in the plasma membrane--typical of apoptosis. TEM indicated that plasma membranes and double nuclear membranes have structural changes, with apoptotic bodies (blebbing) and disrupted mitochondrial membranes. In 17alpha-ethynylestradiol-exposed neurons, disruption of the plasma membrane with cell swelling and vacuolization was present. No apoptotic bodies or budding were noted here. 17beta-Estradiol induced openings in the plasma membrane, while DES-exposed neurons did not show any morphological changes. Therefore we conclude that EDC damage is morphologically visible and the damage is recognized as apoptosis and oncosis. Estrogenic substances may hence modify hormonal actions thereby leaving the developing nervous system more susceptible to damaging events. PMID- 17075773 TI - Glucose-induced effects and joker function of glucose: endocrine or genotoxic prevalence? AB - The steady increase in chronic "glycemic load" is characteristic for modern times. Among myriad of glucose functions, two principals can be emphasized: first, endocrine (in particular, ability to induce insulin secretion) and second, DNA-damaging related to formation of reactive oxygen species (ROS). It was suggested by us earlier that a shift in the ratio of mentioned functions reflects a possible "joker" role of glucose as an important modifier of human pathology. Therefore, we embarked on a study to investigate an individual effect of peroral glucose challenge on serum insulin level and ROS generation by mononuclears (luminol-dependent/latex-induced chemiluminescence) in 20 healthy people aged between 28-75. Concentrations of glucose, blood lipids, carbonylated proteins, malondialdehyde, leptin and TNF-alpha were determined as well. On the basis of received data two separate groups could be distinguished: one (n=8), in which glucose stimulation of ROS generation by mononuclears was increased and relatively prevailed over induction of insulin secretion (state of the so called glucose-induced genotoxicity, GIGT), and another (n=12), in which signs of GIGT were not revealed. People who belonged to the first group were characterized with a tendency to lower body mass index, blood leptin and cholesterol and to higher TNF-alpha concentration. Thus, if joker function of glucose is realized in "genotoxic mode", the phenotype (and probably genotype) of subjects may be rather distinctive to the one discovered in glucose-induced "endocrine prevalence". Whether such changes may serve as a pro-mutagenic or pro-endocrine basis for the rise of different chronic diseases or, rather, different features/aggressiveness of the same disease warrants further study. PMID- 17075774 TI - Effects of growth hormone treatment on B-type natriuretic peptide as a marker of heart failure in adults with growth hormone deficiency. AB - OBJECTIVE: Patients with growth hormone deficiency (GHD) have abnormalities of cardiac structure and function. Growth hormone replacement (GHR) therapy can induce an increase in cardiac mass and improvement in left ventricular ejection fraction. B-type natriuretic peptide (BNP) levels have been successfully used to identify patients with heart failure and they correlate with both disease severity and prognosis. DESIGN: To investigate the effect of growth hormone replacement on BNP and inflammatory cardiovascular risk factors in adults with GHD we determined NT-proBNP and high sensitive C-reactive protein (CrP) before, 6 and 12 months after GHR. PATIENTS: Thirty adults (14 males, 16 females) with GHD mean age: 41.7+/-14.5 years (range: 17.2 to 75.4 years) were recruited from the German KIMS cohort (Pfizer's International Metabolic Database). RESULTS: During 12 months of GHR, a significant increase of IGF-1 (85.4+/-72.1 VS. 172.0+/-98 mug/dl; p=0.0001; IGF-1 SDS mean+/-SD: -3.85+/-3.09 VS. -0.92+/-1.82) was detectable. Mean baseline NT-proBNP was 112+/-130 pg/ml (range: 7 to 562). Twelve patients had normal BNP, whereas 18 revealed NT-proBNP values corresponding to those of patients with heart failure NYHA classification I (n=10), NYHA II (n=6) and NYHA III (n=2), respectively. Baseline BNP levels correlated significantly (p=0.044) with increased baseline CrP values. After 12 months of GHR, a significant decrease (p=0.001) in NT-proBNP levels mean: 68+/-81 pg/ml (range: 5 to 395) was detectable, associated with an improvement in NYHA performance status in 10 of the 18 with increased baseline NT-proBNP. CONCLUSIONS: Based on our study, approximately two-thirds of patients with GHD have increased NT-proBNP levels which may be useful as screening/diagnostic laboratory parameter for heart failure in such patients. GHR therapy decreases BNP levels in most patients with GHD. PMID- 17075775 TI - Cytology and mRNA expression analysis of fine needle aspirates of thyroid nodules in an East German region with borderline iodine deficiency. AB - Fine needle aspiration cytology (FNAC) is widely recommended as an important tool for pre-operative identification of malignancy in patients with nodular thyroid disease. To assess the diagnostic contribution of FNAC and the potential of quantitative mRNA analysis in fine needle aspirates in daily practice, we conducted a prospective study in thyroid clinics (n=2) and endocrine practices (n=3), respectively in an East German region with borderline iodine deficiency. Two-hundred and forty-four consecutive FNACs were obtained over a period of 2 years (2002-2004) from euthyroid patients presenting for first evaluation of a solitary thyroid nodule. The mean nodule size for FNAC was 27 mm (range: 10-79 mm). In 55% of patients FNAC was performed after scintiscan detection of a cold or normal functioning thyroid nodule (CTN), while in the remainder FNAC was performed as a primary investigation. FNAC outcomes were: 57.8% benign, 22.1% indeterminate, 2.5% suspicious for malignancy, 17.6% non-diagnostic. Messenger RNA levels for a house keeping gene (beta-actin) and a thyroid specific marker (thyroglobulin, Tg) were studied as basic molecular markers using real-time PCR. Both in the IN VIVO and EX VIVO FNA series, beta-actin and Tg mRNA levels were positively correlated with the thyrocyte cell yield/respective FNA smear. However, subgroup analysis showed that FNAC with histologically confirmed follicular thyroid cancer and/or microfollicular adenoma exhibited significantly lower Tg mRNA expression despite high beta-actin levels. Sufficient mRNA quantities were obtained in >90% of FNA specimen to allow quantitative mRNA analysis of at least 5 further genes. In conclusion, quantitative mRNA analysis is feasible in FNA on a routine basis and provides a perspective for a molecular distinction of thyroid nodules, once specific marker genes have been defined for benign and malignant thyroid tumours respectively. PMID- 17075776 TI - Metabolic factors affecting residual beta cell function assessed by C-peptide secretion in patients with newly diagnosed type 1 diabetes. AB - BACKGROUND: In recent onset of type 1 diabetes, the residual beta cell function, assessed by baseline and/or stimulated C-peptide secretion, can be a useful parameter to establish the extension of beta cell destruction. How metabolic parameters at diagnosis influence residual C-peptide secretion is not well established. PATIENTS AND METHODS: We analyzed 553 consecutive patients with recent onset (<4 weeks) of type 1 diabetes (250 females and 303 males, mean age 15+/-8 years). Baseline and stimulated C-peptide by i.v. glucagon were evaluated using a highly sensitive radio-immunoassay. Metabolic parameters including blood glucose, HbA1c, insulin dose, and BMI were also evaluated. RESULTS: Baseline and stimulated C-peptide were 0.26+/-0.22 and 0.47+/-0.38 nmol/l and correlated positively with age (p<0.001). There was no significant correlation between C peptide and blood glucose at diagnosis. BMI was positively correlated with both baseline and stimulated C-peptide secretion (p<0.001). By contrast, HbA1c levels inversely correlated with both baseline and stimulated C-peptide secretion (p<0.001). CONCLUSION: In type 1 diabetes at diagnosis, baseline and stimulated C peptide are higher in pubertal and young adult patients compared with pre pubertal patients suggesting that such parameter can be used as an end point marker for studies aimed at protecting and/or restoring beta cells in patients with substantial beta cell function. High levels of HbA1c and lower BMI are dependent variables of C-peptide values. PMID- 17075777 TI - Blood-soluble Fas levels are increased in type 2 diabetic patients with peripheral vascular disease. AB - AIM: To investigate the possible utility of plasma sFas (soluble Fas) levels as a marker of peripheral vascular disease (PVD) in type 2 diabetic patients, and the relationship between classical cardiovascular risk factors and sFas levels in these patients. MATERIALS AND METHODS: sFas levels were measured in 57 type 2 diabetic patients with and 60 without PVD matched for age and sex. Diagnosis of PVD was established in presence of at least one of the following criteria: leg or foot amputation of vascular cause, lower-extremity arterial angioplasty or surgical by-pass, or ankle-braquial index (ABI) less than 1 in at least one side of the body. ELISA was used to measure sFas levels. RESULTS: None of the risk factors assessed total cholesterol, HDL cholesterol, triglycerides, CRP, ACE, fibrinogen, Lp(a) and homocysteine was significantly different between both groups of patients. However, patients with PVD had higher plasma sFas levels than the group without PVD (10.25+/-3.7 ng/ml VS. 8.86+/-2.6 ng/ml; p=0.02). Levels of sFas were 1.45 ng/ml (95% CI: 0.32-2.58; p=0.013) higher in PVD patients when adjusting by age, total, HDL and LDL cholesterol, triglycerides, homocysteine, CRP, ACE, arterial hypertension and tobacco smoking. Using multiple logistic regression sFas is a predictor of PVD, although not potent. CONCLUSION: Plasma sFas may be an independent marker of PVD in type 2 diabetes mellitus patients. PMID- 17075778 TI - Vaso-occlusion in Schimke-immuno-osseous dysplasia: is the NO pathway involved? AB - Schimke-immuno-osseous dysplasia (SIOD) is an autosomal recessive disorder with the main clinical findings of spondyloepiphyseal dysplasia, nephrotic syndrome, and defective cellular immunity. Vaso-occlusive processes, especially generalized atherosclerosis, are a life-limiting complication in patients with severe SIOD. The nitric oxide synthase (NOS) oxidizes L-arginine to nitric oxide (NO). NO is a potent vasodilator with inhibitory effects on platelet aggregation and the development of atherosclerosis. We hypothesized that reduced NO production due to antagonism of NOS by asymmetric dimethylarginine (ADMA) would be a possible pathophysiological mechanism for vaso-occlusion in SIOD. We tested this hypothesis in 10 patients with SIOD and 10 age-matched healthy controls. Plasma and urine levels of nitrite and nitrate, the indicators of NO synthesis, and of ADMA, an endogenous NOS inhibitor, in children suffering from SIOD were not significantly different from those in the age-matched healthy controls. Our results suggest that the L-arginine/NO pathway is not altered in SIOD. Antagonism of NOS by ADMA does not seem to be the cause of premature general atherosclerosis in SIOD. The underlying pathology of vaso-occlusion in SIOD still remains unclear. PMID- 17075779 TI - Regular insulin secretory oscillations despite impaired ATP synthesis in Friedreich Ataxia patients. AB - Friedreich Ataxia is an inherited disorder caused by decreased expression of a mitochondrial protein called frataxin. Deficiency of this protein causes reduced biogenesis of iron-sulfur clusters, and subsequently impaired synthesis and replenishment of ATP IN VIVO. Basal secretion of insulin occurs in an oscillating manner presumably triggered by ATP-dependent feedback inhibition of glycolytic flux. Hence, individuals with reduced ATP synthesis rates should possibly exhibit impaired insulin secretory oscillations if these were solely dependent on ATP. In the present study Friedreich Ataxia patients with a presumptive impairment of ATP synthesis in pancreatic beta-cells were evaluated for regularity of basal secretory oscillations of insulin. Healthy siblings were employed as controls. In conflict with the initial hypothesis, no differences in regards to oscillation patterns were observed between patients and controls. Supported by EX VIVO evidence, these findings tentatively suggest that pulsatile insulin secretion might not be exclusively dependent on ATP feedback inhibition in humans. PMID- 17075780 TI - Soluble TNF receptor and periodontal disease in non-obese Japanese type 2 diabetic patients. PMID- 17075781 TI - Biguanides enhance glucose utilization and insulin action in peripheral tissues: an old finding from the sixties of the last century. PMID- 17075782 TI - [Recent advances in prenatal diagnostics]. AB - During the last years, technical improvements have increased the possibilities in prenatal ultrasound. During the eighties and nineties, fetal malformations were increasingly detected and specified. Since a few years, the measurement of the fetal nuchal translucency between 11 and 14 weeks of gestation has been implemented to calculate the individual risk, in combination with most recent biochemical markers. Today, the sonographic measurement of the nuchal translucency is regarded as a valuable screening tool for chromosomal anomalies in prenatal medicine. Beside standardized examinations, a profound information and counseling of the pregnant women should be emphasized. With the improvement of the specific maternal risk calculation, using the sonographic measurement of the nuchal translucency, the biochemical markers and the maternal age, unnecessary invasive examinations may be prevented and their overall number can significantly be reduced. The same trend is seen in the whole field of prenatal medicine, illustrated by the detection of the fetal rhesus D status from the maternal blood and the use of Doppler ultrasound in the management of fetal anemia. PMID- 17075783 TI - [First-trimester-test--support-tool for the individual decision or screening-tool for public health planning?]. AB - International studies about women's knowledge of the first-trimester-test show that they are quite often not sufficiently informed for their personal decision making: The information needed is not given in an understandable way; they are not informed that the test is only a risk assessment and not a diagnosis; and they often don't understand the concept of false positive and false negative results. Other studies show, that this sophisticated and complex information about the first-trimester-test can be given in an understandable way. But even with adequate information most women are unable to make an informed choice as long as the test is presented as a routine for screening. However, for an informed and free choice, an individual decision-making-process has to take place. For this reason a counseling concept has been developed in Switzerland. It introduces the first-trimester-test not as a screening-tool for the public health planning but as a support-tool for the individual decision. It consists of an information brochure for the pregnant woman and her partner, a counseling framework for the physicians and a two days training program. The counseling concept has been evaluated by a research project of the Swiss National Science Foundation and has been adopted as an official counseling standard by the Swiss Society of Obstetrics and Gynaecology. Recommendations for the communication skills together with a short time training program were added, such that the concept can serve as an integrative tool for decision-making support concerning first-trimester tests. PMID- 17075784 TI - [Is informed consent actually possible?]. AB - In prenatal medicine, the doctrine of informed consent is subject to several restrictions: women are confronted with social expectations to accept screening and tests that entail a pressure to decide. Some authors criticise that the informed consent overstretches the patient, dislocates responsibility to the individual, and, in many cases, is nothing but an empty ritual. The article defends the idea of informed consent. It argues that we need to reinterpret informed consent on the basis of a dialogical principle: the aim is to recognise the subjectivity and vulnerability of the patient in her special situation, which implies a mutual culture of hearing. PMID- 17075785 TI - [New options in prenatal and preimplantation diagnosis of genetic disorders]. AB - During recent years the progress with the most important practical impact in prenatal diagnosis has been the implementation of first trimester risk screening for common aneuploidies leading to a much improved identification of pregnancies at risk. Molecular methods for a rapid, cost-effective, but selective aneuploidy diagnosis such as interphase FISH or QF-PCR have been around for years, do have their specific indications, but will unlikely replace conventional cytogenetic tools in routine diagnosis. They apparently do also play a role as marketing instruments in the competition among cytogenetic laboratories. The most thrilling issue for all cytogeneticists in the years to come will be the introduction of array-based methods in the prenatal routine diagnosis of chromosomal abnormalities. Polar body diagnosis has been the only option available for preimplantation genetic diagnosis in german speaking countries. The overwhelming majority of all professionals involved and many families concerned share the hope that the legal situation will improve in these countries to allow the examination of early embryos in high risk situations. PMID- 17075786 TI - [Ethical and biopolitical problems of the stem cell research]. AB - Stem cells have offered much hope by promising to greatly extend the numbers and range of patients who could benefit from transplants, and to provide cell replacement therapy to treat diseases such as diabetes, Parkinson's and Huntington's disease. The issue of stem cell research is politically charged, prompting biologists to begin engaging in ethical debates, and generating in the general public an unusually high level of interest in this aspect of modern biology and medicine. But excitement notwithstanding, there is a long way to go in basic research before new therapies will be established, and now the pressure is on for scientists and clinicians to deliver. PMID- 17075787 TI - [Is it possible to answer the question about the beginning of human life?]. AB - The question of the beginning of human life is specified in this article by explaining the different meanings of the expression "human life". Then it is explained that every discussion about this beginning is always contextual and thus one can detect different "beginnings" of human life. Therefore the question raised into the title can only find part answer and none branch neither in the field of exact sciences nor in the field of humanities can claim to have a position of monopole. Finally, one tries to demonstrate that even if the question of the beginning of life could find an answer, the normative conflicts that would appear at that moment, wouldn't automatically be solved, nor settled. PMID- 17075788 TI - [Human embryonic stem cell therapy, what is the attitude of the public?]. AB - This work discusses the manner in which the public has dealt with the ethical questions about human embryonic stem cell research. It argues that the public is not a homogenous entity, but rather consists of a number of "different publics", which at different points of the debate have discussed different aspects and different questions of this therapeutic proposal. It understands the development of human stem cell therapy not as a result of the activity of an isolated science, acting independently of society. In the contrary it considers it as a co production of science with the different social actors, which participate in the political, ethical, and legal processes. Therefore the question if ethics is ahead or lags behind the developments in medical research becomes obsolete. Ethics is on one side the result of the effort to handle the results of biomedical research in a manner which is adequate for society at large on the other side it also influences the scientific-technological development through its own assessment. PMID- 17075789 TI - [Ethical considerations in the management of infants born at the limit of viability]. AB - Despite ongoing progress in perinatal care over the past decade, mortality rates of infants born before 24 completed weeks of gestation have remained high, and the majority of survivors have at least some degree of neurosensory impairment. With increasing knowledge of long-term follow-up data, quality of life aspects have become more important in treatment decisions for infants born at the limit of viability. Many countries have adopted an individualized approach to the care of these infants. Provisional intensive care is initiated in the delivery room and continued in the neonatal intensive care unit as long as there is a reasonable chance of survival and the expected quality of life appears to justify the patient's pain and suffering. On the other hand, redirection of care becomes an ethically justifiable option once the burdens have begun to outweigh the benefits. The published recommendations from different countries show considerable variability. For example, the gestational age below which preference should be given to palliative care ranges between 22 (Germany, Japan) and 25 completed weeks of gestation (Netherlands). Similarly, parental participation in surrogate decision making varies from country to country. Recommendations which emphasize quality of life aspects tend to encourage parental participation more than recommendations which are primarily based on a sanctity of life ideology. The quest to improve care for infants born at the limit of viability will continue. Trying to push the limit of viability towards even lower gestational ages is not a priority. Research efforts should focus on improving long-term outcome for survivors and on developing high quality palliative care for non survivors. PMID- 17075792 TI - Intracranial therapy of glioblastoma with the fusion protein DTAT in immunodeficient mice. AB - A gene splicing technique was used to create a hybrid fusion protein DTAT encoding the 390 amino acid portion of diphtheria toxin (DT(390)), a linker, and the downstream 135-amino terminal fragment portion of human urokinase plasminogen activator. DTAT was assembled to target human glioblastoma cell lines in a murine intracranial model. Previously published in vitro studies demonstrated that DTAT was highly selective and toxic to human glioblastoma cell lines in a flank tumor model. The purpose of this study was to determine the toxicity, specificity and possible therapeutic efficacy of DTAT in an intracranial model. Convection enhanced delivery of DTAT resulted in about a 16-fold increase in maximum tolerated dose. Intracranial administration of DTAT on an every-other-day basis in nude mice with established U87 MG brain tumors resulted in significant reductions in tumor volume and significantly prolonged survival (p < 0.0001). Magnetic resonance imaging proved to be a powerful tool in mice and rats for demonstrating tumor growth in a xenograft intracranial model, assessing the efficacy of DTAT in tumor volume reduction and detecting DTAT-associated intracranial toxicity and vascular damage. These results suggest that the DTAT recombinant fusion protein is highly effective in an intracranial model and DTAT might be an effective treatment for glioblastoma. PMID- 17075793 TI - Serum PAPP-A levels at 10-14 weeks of gestation are altered in women after assisted conception. AB - OBJECTIVES: Increased screen-positive rate was observed in the first-trimester screening for trisomy 21 (T21) among women after assisted conception. The aim of this study was to identify the trends of changes in serum marker levels after different modes of assisted conception. METHODS: Retrospectively, we analyzed the levels of free beta human chorionic gonadotrophin (f beta HCG), pregnancy associated plasma protein-A (PAPP-A) and inhibin A in maternal serum, and fetal nuchal translucency (NT) thickness at 10-14 gestational weeks in 1098 women with singleton pregnancies: 130 after IVF-ET, 54 after ICSI and 914 after spontaneous conception. RESULTS: In women after IVF-ET and ICSI, PAPP-A was decreased (0.94 MoM and 0.82 MoM, respectively) and inhibin A increased (1.11 MoM and 1.48 MoM, respectively) in comparison to the women after spontaneous conception. With increasing number of oocytes retrieved PAPP-A decreased and inhibin A increased. In women pregnant after assisted conception without ovarian stimulation (transfer of frozen-thawed embryo or in spontaneous cycle) the marker levels were not significantly different from those in women after spontaneous conception. CONCLUSIONS: The presence of multiple corpora lutea may be responsible for the changes in marker levels. In spite of the differences in marker levels, the screen-positive rate for T21 is not significantly increased in women after assisted conception. PMID- 17075794 TI - Cytogenetic and histological features of a human embryo with homogeneous chromosome 8 trisomy. AB - BACKGROUND: Homogeneous and complete trisomy 8 is extremely rare. With one recent neonatal exception, all reported cases have been mosaic, due to mitotic non disjunction during early zygotic development. We report a case of chromosome 8 trisomy in a human embryo examined at Carnegie stage 11 (25 days post fertilization). It presented severe cardiovascular and central nervous system malformations. METHODS: The unusual bifid heart in this embryo spurred a detailed histological examination, karyotyping of a chorionic villus sample and subsequent FISH on inter-phase nuclei of intra-embryonic sections. RESULTS: Trophoblast cells had a karyotype of 47,XX, +8. Within the embryo proper, FISH demonstrated that the trisomy 8 was homogeneous in embryonic as well as extra-embryonic tissues. FQ-PCR supports a meiosis I origin of non-disjunction. In sections, the pharyngeal arches (including cardiac outflow tract), forebrain, mesonephros and liver were absent. Somites and yolk sac blood vessels were irregularly shaped. CONCLUSION: We show that homogeneous, intra-embryonic trisomy 8 is compatible with implantation and early human development. Molecular pathways that may be compromised and their impact on organogenesis are discussed. PMID- 17075795 TI - Mosaic trisomy 16 in a fetus: the complex relationship between phenotype and genetic mechanisms. AB - OBJECTIVES: This study was undertaken to discuss the workup of trisomy 16 pregnancies. STUDY DESIGN: This case study reports the prenatal detection and postnatal confirmation of mosaic trisomy 16, associated with uniparental disomy (UPD) 16, in a 34-year-old woman who showed elevated maternal serum alpha fetoprotein and beta-HCG at a gestational age (GA) of 15.5 weeks. RESULTS: Amniotic fluid (AF) karyotyping at different GAs revealed various levels of trisomy 16 mosaicism (0 to level III). UPD studies at 21 weeks of gestation revealed maternal heterodisomy 16. Serial fetal ultrasonography showed fetal abnormalities: intrauterine growth restriction (IUGR), dilated digestive tract, and gallbladder agenesis. Postmortem examination confirmed the prenatal findings and revealed additional anomalies, such as hypoplastic cerebellum with abnormal gyration of the vermis. CONCLUSIONS: Workup following prenatal detection of trisomy 16 mosaicism in chorionic villi must include AF karyotyping and serial ultrasound examination of the fetus in order to approach postnatal developmental prognosis. PMID- 17075796 TI - Large-scale chromatin remodeling in germinal vesicle bovine oocytes: interplay with gap junction functionality and developmental competence. AB - In mammals, oocyte acquires a series of competencies sequentially during folliculogenesis that play critical roles at fertilization and early stages of embryonic development. In mouse, chromatin in germinal vesicle (GV) undergoes dynamic changes during oocyte growth and its progressive condensation has been related to the achievement of developmental potential. Cumulus cells are essential for the acquisition of meiotic competence and play a role in chromatin remodeling during oocyte growth. This study is aimed to characterize the chromatin configuration of growing and fully grown bovine oocytes, the status of communications between oocyte and cumulus cells and oocyte developmental potential. Following nuclear staining, we identified four discrete stages of GV, characterized by an increase of chromatin condensation. GV0 stage represented 82% of growing oocytes and it was absent in fully grown oocytes. GV1, GV2, and GV3 represented, respectively, 24, 31, and 45% of fully grown oocytes. Our data indicated a moderate but significant increase in oocyte diameter between GV0 and GV3 stage. By dye coupling assay the 98% of GV0 oocytes showed fully open communications while the number of oocytes with functionally closed communications with cumulus cells was significantly higher in GV3 group than GV1 and GV2. However, GV0 oocytes were unable to progress through metaphase II while GV2 and GV3 showed the highest developmental capability. We conclude that in bovine, the progressive chromatin condensation is related to the sequential achievement of meiotic and embryonic developmental competencies during oocyte growth and differentiation. Moreover, gap-junction-mediated communications between oocyte and cumulus cells could be implicated in modulating the chromatin remodeling process. PMID- 17075797 TI - The mRNA expression of P450 aromatase, gonadotropin beta-subunits and FTZ-F1 in the orange-spotted grouper (Epinephelus Coioides) during 17alpha methyltestosterone-induced precocious sex change. AB - The orange-spotted grouper Epinephelus coioides is a protogynous hermaphroditic fish, but the physiological basis of its sex change remains largely unknown. In the present study, the 2-year-old orange-spotted grouper was induced to change sex precociously by oral administration of 17alpha-methyltestosterone (MT, 50 mg/Kg diet, twice a day at daily ration of 5% bodyweight) for 60 days. The serum testosterone levels were significantly elevated after MT treatment for 20 and 40 days as compared to control, but the levels of serum estradiol (E(2)) remained unchanged. The expression of P450aromA in the gonad significantly decreased after MT treatment for 20, 40, and 60 days. Accordingly, the enzyme activity of gonadal aromatase was also lower. The expression of FSHbeta subunit in the pituitary was significantly decreased after MT treatment for 20 days, but returned to the control levels after 40 and 60 days; however, the expression of LHbeta subunit was not altered significantly by MT treatment. The expression of FTZ-F1 in the gonad also decreased significantly in response to MT treatment for 40 and 60 days, but its expression in the pituitary was not altered significantly. Interestingly, when tested in vitro on ovarian fragments, MT had no direct effect on the expression of P450aromA and FTZ-F1 as well as the activity of gonadal aromatase, suggesting that the inhibition of gonadal P450aromatase and FTZ-F1 by MT may be mediated at upper levels of the brain-pituitary-gonadal axis. Taken together, these results indicated that FSH, P450aromA, FTZ-F1, and serum testosterone are associated with the MT-induced sex change of the orange-spotted grouper, but the cause-effect relationship between these factors and sex change in this species remains to be characterized. PMID- 17075798 TI - Characterization of circulating blood dendritic cell subsets DC123+ (lymphoid) and DC11C+ (myeloid) in prostate adenocarcinoma patients. AB - PURPOSE: We verified whether prostate adenocarcinoma produces specific modifications in DC subsets count. METHODS: Twenty-one untreated prostate adenocarcinomas were divided on the basis of clinical stage in localized and metastatic disease. As control we used a population of 18 healthy male subjects. For DCs enumeration, peripheral blood (PB) samples were obtained in all cases. A single-platform flow cytometric assay based on Tru-COUNT was used for the enumeration of the two DCs subsets, myeloid (mDCs) and plasmacytoid (pDCs). RESULTS: We showed a statistically significant reduction in pDCs count in prostate cancer population when compared to healthy controls (P = 0.002). Comparing each clinical stage with healthy controls, significant differences were found between controls and the metastatic group in both pDCs and mDCs (P = 0.005 and P = 0.023 respectively) but not between controls and the localized group (P = 0.055 and P = 0.829 respectively). CONCLUSIONS: We showed that DCs count in PB is significantly affected by prostate adenocarcinoma progression in a metastatic disease. PMID- 17075799 TI - Sulfasalazine-induced cystine starvation: potential use for prostate cancer therapy. AB - BACKGROUND: Certain cancers depend for growth on uptake of cystine/cysteine from their environment. Here we examined advanced human prostate cancer cell lines, DU 145 and PC-3, for dependence on extracellular cystine and sensitivity to sulfasalazine (SASP), a potent inhibitor of the x(c)(-) cystine transporter. METHODS: Cultures were evaluated for growth dependence on exogenous cystine, x(c)(-) transporter expression, response to SASP (growth and glutathione content). In vivo, effect of SASP was determined on subrenal capsule xenograft growth. RESULTS: Cystine omission from culture medium arrested DU-145 and PC-3 cell proliferation; both cell lines expressed the x(c)(-) transporter and were growth inhibited by SASP (IC(50)s: 0.20 and 0.28 mM, respectively). SASP-induced growth inhibition was associated with vast reductions in cellular glutathione content - both effects based on cystine starvation. SASP (i.p.) markedly inhibited growth of DU-145 and PC-3 xenografts without major toxicity to hosts. CONCLUSIONS: SASP-induced cystine/cysteine starvation leading to glutathione depletion may be useful for therapy of prostate cancers dependent on extracellular cystine. PMID- 17075800 TI - Magnetic resonance imaging of phase transitions in nitinol. AB - Magnetic resonance images are prone to artifacts caused by metallic objects. Apart from being a source of image degradation, such artifacts can also provide information about the magnetic properties of the foreign object. In this work, we aim to explore the potential of magnetic resonance imaging to detect and characterize changes in magnetic properties of nitinol undergoing temperature- or strain-induced phase changes. A spin echo and a gradient echo method were used to measure the magnetization changes related to the phase transformations. Results of both methods were in agreement and in accordance with the independent measurements using a vibrating sample magnetometer. Magnetic resonance imaging turned out to be a suitable method to visualize and quantify magnetization and phase changes in situ. It is not restricted to a single imaging strategy and does not require any modification of the test object. The results indicate the potential of magnetic resonance imaging to provide direct feedback of the thermomechanical state of the alloy. PMID- 17075801 TI - IK cytokine ameliorates the progression of lupus nephritis in MRL/lpr mice. AB - OBJECTIVE: IK cytokine has been isolated as a factor that inhibits interferon gamma (IFNgamma)-induced expression of class II major histocompatibility complex (MHC) antigens. Aberrant expression of class II MHC antigens has reportedly been recognized in the target organs of autoimmune diseases and been associated with disease activity. In this study, we investigated whether IK cytokine can ameliorate the progression of lupus nephritis in MRL/lpr mice. METHODS: A truncated IK analog was prepared and transfected into a nonmetastatic fibroblastoid cell line, and then injected subcutaneously into MRL/lpr mice at ages 8 weeks (before the onset of lupus nephritis) and 12 weeks (at the early stage of the disease). RESULTS: An IK cytokine, when it was translated from methionine at position 316, acted as a secretory protein. This truncated IK cytokine (tIK) reduced IFNgamma-induced class II MHC expression in various cells through decreased expression of class II MHC transcription activator. Treatment of MRL/lpr mice with tIK significantly reduced renal damage as compared with control mice. A significant decrease in macrophage and T cell infiltration was found in the kidneys of tIK-treated mice, resulting in decreased production of IFNgamma and interleukin-2. Mice treated with tIK also showed significant reduction of anti-DNA antibodies and circulating immune complexes. A specific reduction of class II MHC expression was observed on B cells and monocytes as well as in the kidney. CONCLUSION: We prepared a potent IK analog and demonstrated its ability to ameliorate the progression of lupus nephritis. This agent may therefore provide a new therapeutic approach for lupus nephritis. PMID- 17075802 TI - Glycyrrhizin. PMID- 17075805 TI - Expansion of toll-like receptor 9-expressing B cells in active systemic lupus erythematosus: implications for the induction and maintenance of the autoimmune process. AB - OBJECTIVE: Toll-like receptors (TLRs) are pattern-associated receptors in innate immunity that may be involved in the recognition of self antigens and the production of pathogenic autoantibodies. This study was undertaken to examine the expression and function of various TLRs in subpopulations of peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE). METHODS: The expression of TLRs in PBMCs from 50 SLE patients with active disease (SLE Disease Activity Index [SLEDAI] score >or=8; n = 26) or inactive disease (SLEDAI score <8; n = 24) and 20 healthy controls was studied by flow cytometry. TLR expression was assessed on various subpopulations of PBMCs (TLR-2 and TLR-4 by membrane staining; TLR-3 and TLR-9 by intracellular staining). TLR function was accessed by stimulating PBMCs with specific ligands. RESULTS: The proportion of B cells and monocytes expressing TLR-9 was higher among patients with active SLE (mean +/- SD 49.5 +/- 24.4% and 30.7 +/- 24.1%, respectively) than among patients with inactive disease (22.8 +/- 19.6% and 14.3 +/- 8.4%, respectively; P = 0.02 and P = 0.03). Among B cells, the proportion of plasma cells and memory B cells expressing TLR-9 was increased in patients with active SLE. Increased percentages of TLR-9-expressing B cells correlated with the presence of anti double-stranded DNA antibodies (P = 0.007). Treatment with serum from patients with active disease increased the percentage of TLR-9-expressing plasma cells in serum from healthy controls. Enhanced induction of HLA-DR after TLR-9 stimulation was documented in B cells from patients with active disease. CONCLUSION: In patients with active SLE, the proportion of peripheral blood memory B cells and plasma cells expressing TLR-9 is increased. Endogenous nucleic acids released during apoptotic cell death may stimulate B cells via TLR-9 and contribute to SLE pathogenesis. PMID- 17075806 TI - B cell depletion therapy in systemic lupus erythematosus: effect on autoantibody and antimicrobial antibody profiles. AB - OBJECTIVE: Autoantibody production in patients with systemic lupus erythematosus (SLE) is associated with abnormalities of B cell function and phenotype. Clinical responses to B cell depletion therapy (BCDT), based on rituximab, are encouraging. Therefore, we undertook this study to investigate the effect of BCDT on antibody profiles. METHODS: Serial sera from 16 patients with active, refractory SLE were assayed for antinucleosome antibodies, anti-double-stranded DNA (anti-dsDNA), anti-extractable nuclear antigen, anti-tetanus toxoid, and antibodies to pneumococcal capsular polysaccharide for at least 1 year following BCDT. Anti-dsDNA antibodies derived from the V(H)4.34 immunoglobulin germ line gene (9G4+) were also measured. RESULTS: All patients achieved peripheral B cell depletion and improved clinically for at least 3 months. Antinucleosome and anti dsDNA antibodies decreased to a mean +/- SD of 64 +/- 37% and 38 +/- 33% of baseline values, respectively, by 6-8 months post-BCDT. Levels of other autoantibodies and antimicrobial antibodies were generally unchanged. In the 9 of 16 patients who were still well at 1 year, anti-dsDNA antibodies fell to 42 +/- 36% of baseline values at 6-8 months and to 37 +/- 33% at 10-14 months. In patients who had disease flares within 1 year of BCDT, levels of these antibodies decreased to 60 +/- 40% and 83 +/- 93% of baseline values at 6-8 months and at 10 14 months, respectively. Circulating anti-dsDNA antibodies were positive for 9G4 expression in 4 of 6 patients tested, and flares in 2 of these patients were accompanied by rises in 9G4+ anti-dsDNA antibodies. CONCLUSION: These observations suggest that B cell clones committed to producing antinucleosome and anti-dsDNA antibodies, including the V(H)4.34 subpopulation of anti-dsDNA antibodies, have a relatively rapid turnover compared with B cell clones producing other antibodies. There was also a trend toward a greater and more sustained decrease in anti-dsDNA antibodies in patients with clinical benefit lasting >1 year. PMID- 17075807 TI - The effect of moderate-dose corticosteroids in preventing severe flares in patients with serologically active, but clinically stable, systemic lupus erythematosus: findings of a prospective, randomized, double-blind, placebo controlled trial. AB - OBJECTIVE: Serial measurements of anti-double-stranded DNA (anti-dsDNA) and complement are routine in the management of systemic lupus erythematosus (SLE), but their utility as biomarkers in preemptive treatment to prevent flares remains a subject of controversy. We hypothesized that concomitant elevation of anti dsDNA and C3a can predict SLE activity in patients with stable or inactive disease and that short-term treatment with corticosteroids can avert flares. METHODS: In this prospective, randomized, double-blind, placebo-controlled trial, 154 patients were evaluated monthly for up to 18 months, with measurements of C3a, C3, C4, CH50, and anti-dsDNA levels. Patients who remained clinically stable but showed serologic evidence of an SLE flare (elevation of both the anti-dsDNA level by 25% and the C3a level by 50% over the previous 1-2 monthly visits) were randomized to receive either prednisone or placebo therapy at a dosage of 30 mg/day for 2 weeks, 20 mg/day for 1 week, and 10 mg/day for 1 week. RESULTS: Forty-one patients (21 randomized to prednisone and 20 randomized to placebo) experienced a serologic flare. Analysis of severe flares occurring 40 mg/day and/or the addition of an immunosuppressive agent. Furthermore, improvement in scores on the Systemic Lupus Erythematosus Disease Activity Index, decreased levels of anti-dsDNA antibodies, and increased levels of C4 occurred 1 month after initiation of prednisone treatment. CONCLUSION: These preliminary data support our hypothesis that in a subset of clinically stable SLE patients with a combination of elevated C3a and anti-dsDNA levels, short-term corticosteroid therapy may avert a severe flare. PMID- 17075808 TI - Hepatocyte growth factor and transforming growth factor beta1 ratio at baseline can predict early response to cyclophosphamide in systemic lupus erythematosus nephritis. AB - OBJECTIVE: To determine whether the ratio of hepatocyte growth factor (HGF) to transforming growth factor beta1 (TGFbeta1) in systemic lupus erythematosus (SLE) nephritis could be a prognostic factor for response to therapy with cyclophosphamide (CYC) and steroids at 6 months, and to examine whether the molecular ratio of HGF to TGFbeta1 correlates with the activity index (AI) and chronicity index (CI) and has predictive value for remission at the sixth month. METHODS: Thirty-six SLE patients with new-onset nephritis, 25 of whom were treated with CYC and steroids, entered into a prospective observational cohort trial at a tertiary university referral center. Renal biopsy findings and clinical parameters were recorded for all patients. Histopathologic, clinical, and immunohistochemical data at baseline served to define the predictive value for the outcome at 6 months. RESULTS: AI and CI at baseline did not distinguish patients who had achieved remission from those who had not achieved remission after receiving CYC plus steroids. HGF and TGFbeta1 were expressed in the tubuli, not in the glomeruli. The CI correlated directly with the TGFbeta1 extension score (TGFbeta1-ES) (r = 0.43, P = 0.008), but correlated indirectly with the HGF intensity score (HGF-IS) (r = -0.39, P = 0.02) and the HGF-ES (r = -0.45, P = 0.006). An HGF-ES:TGFbeta1-ES ratio of >or=1 at baseline distinguished patients who had achieved remission from those who had not achieved remission, with a predictive value of 94%. CONCLUSION: These findings indicate that baseline expression of renal HGF and TGFbeta1 predicts short-term renal outcome after therapy with CYC and steroids. PMID- 17075809 TI - Discrimination between benign and malignant prostate tissue using chromatin texture analysis in 3-D by confocal laser scanning microscopy. AB - BACKGROUND: Analysis of chromatin texture may improve both the diagnosis and the assessment of the prognosis of prostate cancer. Confocal laser scanning microscopy (CLSM) allows performing measurements in nuclei reconstructed in 3-D. The aim of this study was to evaluate the clinical usefulness of 3-D texture analysis of prostate tissue. METHODS: Image stacks of eight prostate cancer sections were obtained by CLSM of both benign and malignant areas. Texture feature values were computed for individual nuclei. The discriminative power of the texture features was established by receiver operating characteristic (ROC) analysis and linear discriminant analysis (LDA). RESULTS: Texture features were identified that could discriminate between benign and malignant nuclei. LDA correctly classified 89% of the nuclei of the pooled set of benign and malignant nuclei. CONCLUSIONS: 3-D nuclear texture features allow discrimination of most benign and malignant prostate nuclei. We estimate that the classification rates can be increased by improving the image quality. PMID- 17075810 TI - Hydroxychloroquine in lupus pregnancy. AB - OBJECTIVE: Hydroxychloroquine (HCQ) is often needed to manage disease activity in systemic lupus erythematosus (SLE) during pregnancy. The purpose of this study was to examine lupus activity and pregnancy outcomes in women with SLE treated or not treated with HCQ during pregnancy. METHODS: This was a prospective study of pregnancies in women with SLE who were evaluated between 1987 and 2002. The pregnancies were divided into 3 groups: no HCQ exposure during pregnancy (163 pregnancies), continuous use of HCQ during pregnancy (56 pregnancies), or cessation of HCQ treatment either in the 3 months prior to or during the first trimester of pregnancy (38 pregnancies). The pregnancy outcomes, fetal outcomes, and lupus activity during pregnancy were compared among these groups. RESULTS: The rates of miscarriage, stillbirth, pregnancy loss, and congenital abnormality were not statistically different among the 3 groups. The degree of lupus activity during pregnancy, however, was significantly higher in women who stopped taking HCQ. These women had a higher degree of lupus activity, as measured by the physician's estimate of lupus activity and the SLE Disease Activity Index, as well as an increased rate of flare, during pregnancy. More serious lupus complications, such as proteinuria and thrombocytopenia, were not significantly higher in women who stopped taking HCQ. Women who continued taking HCQ were maintained on a lower average dose of prednisone during pregnancy. CONCLUSION: We recommend the continuation of HCQ treatment during pregnancy. Our findings are consistent with prior reports of the absence of fetal toxicity. Similar to studies of nonpregnant women, the cessation of HCQ treatment during pregnancy increases the degree of lupus activity. PMID- 17075811 TI - Occupational silica and solvent exposures and risk of systemic lupus erythematosus in urban women. AB - OBJECTIVE: To assess the risk of systemic lupus erythematosus (SLE) associated with occupational exposure to silica dust and organic solvents in an urban population. METHODS: Women with SLE were identified through both community screening and hospital databases in 4 predominantly African American neighborhoods in Boston. Female control patients were volunteers from the same communities and were screened for the absence of connective tissue disease. Demographic factors, smoking history, and a detailed occupational history, including exposures to specific chemicals, were obtained by in-person interviews. The exposure assessment was based on independent evaluation of the occupational history by 2 reviewers who were blinded to each subject's disease status. The risks associated with exposure to silica and solvents were analyzed using multivariate conditional logistic regression models, adjusted for potential confounders. RESULTS: Ninety-five patients and 191 age- and race-matched controls were included in this analysis. Exposure to silica for longer than 1 year was associated with SLE (odds ratio [OR] 4.3, 95% confidence interval [95% CI] 1.7 11.2). An exposure-response effect was seen for longer duration of exposures to silica (P for trend = 0.01). The association between occupational exposure to organic solvents and SLE was not statistically significant (OR 1.04, 95% CI 0.34 3.2). CONCLUSION: Silica exposure from a variety of industrial occupations in urban areas is associated with an increased risk of SLE. A longer duration of exposure to silica dust is associated with greater risks. This study provides further impetus for additional research into the influence of modifiable exposures on the pathogenesis of SLE. PMID- 17075812 TI - Acupuncture for treating osteoarthritis of the knee and the hip. PMID- 17075813 TI - The large print giveth and the small print taketh away: preemptive treatment of serologically active, clinically quiet systemic lupus erythematosus. PMID- 17075814 TI - Myofibroblasts and hyalinized collagen as markers of skin disease in systemic sclerosis. AB - OBJECTIVE: To investigate the correlation between the degree of dermal fibrosis and myofibroblast infiltration using clinical assessments of skin thickness and hardness in systemic sclerosis (SSc). METHODS: Eleven patients with diffuse SSc and 10 healthy controls were evaluated using the modified Rodnan skin thickness score and durometry (hardness measurement). Biopsy samples were obtained from the dorsal mid-forearm in all subjects at the baseline visit and again 6-12 months later in patients with SSc. Five of the patients with SSc received treatment with cyclophosphamide (CYC) in the interval between skin biopsies. Biopsy sections were assessed for myofibroblast and hyalinized collagen content by 2 blinded observers. RESULTS: Myofibroblast and hyalinized collagen scores each correlated with the forearm skin score (r = 0.83, P < 0.0001 and r = 0.78, P < 0.0001, respectively) and with the forearm durometry score (r = 0.72, P < 0.0004 and r = 0.69, P < 0.0008, respectively). The change in the dermal hyalinized collagen score correlated with the change in the forearm durometry score (r = 0.74, P < 0.0213). The myofibroblast score decreased in all 5 patients who received CYC and increased in those receiving non-CYC treatments (P < 0.01 for the difference). CONCLUSION: Myofibroblasts play an important role in the pathogenesis of fibrosis, and our data imply that quantification of myofibroblasts and hyalinized collagen in skin may be a useful outcome measure in clinical studies of SSc. PMID- 17075815 TI - Macrophage migration inhibitory factor promoter polymorphisms and the clinical expression of scleroderma. AB - OBJECTIVE: To investigate the potential association between functional polymorphisms in the gene for the innate mediator, macrophage migration inhibitory factor (MIF), and the clinical expression of systemic sclerosis (SSc). METHODS: Genomic DNA samples and clinical data were collected from the Scleroderma Family Registry and DNA Repository at the University of Texas Health Science Center at Houston. A total of 740 subjects were studied; 203 of them had diffuse cutaneous SSc (dcSSc), 283 had limited cutaneous SSc (lcSSc), and the remaining 254 healthy subjects served as controls. Association analyses were performed on the whole data set and on patient and sex subsets. Significant relationships were determined between clinical variables and MIF polymorphisms for each disease subtype in the studied groups. RESULTS: The frequency of the 173*C MIF allele, which was previously reported to be associated with high production of MIF, was lower in the lcSSc group (12.6%) than in the dcSSc (19.2%) or control (18.5%) groups (P = 0.010 and P = 0.011, respectively). Haplotype analysis for 2 closely linked polymorphisms in the MIF promoter showed that in white subjects with lcSSc or dcSSc, the lcSSc population had a significantly lower representation of the high-expression MIF haplotype defined by -173*C and 794 with 7 CATT repeats (C7) (P = 0.015, odds ratio 1.94 [95% confidence interval 1.14-3.32]). Fibroblasts encoding the C7 MIF haplotype were observed to produce more MIF upon in vitro stimulation than those with a non-C7 haplotype. CONCLUSION: Functional promoter polymorphisms in the MIF gene affect the clinical presentation of SSc. The proinflammatory haplotype defined by C7 is underrepresented in patients with lcSSc. PMID- 17075816 TI - Citrullinated proteins in rheumatoid arthritis: crucial...but not sufficient! PMID- 17075817 TI - Simultaneous assessment of short-term gastrointestinal benefits and cardiovascular risks of selective cyclooxygenase 2 inhibitors and nonselective nonsteroidal antiinflammatory drugs: an instrumental variable analysis. AB - OBJECTIVE: To simultaneously assess the short-term reduction in risk of gastrointestinal (GI) complications and increase in risk of acute myocardial infarction (MI) by celecoxib compared with rofecoxib and several nonselective nonsteroidal antiinflammatory drugs (NSAIDs) using instrumental variable analysis. METHODS: A population of 49,711 Medicare beneficiaries ages 65 years and older who initiated nonselective NSAID or selective cyclooxygenase 2 inhibitor therapy between January 1, 1999, and December 31, 2002, was identified. The increase in risk of GI complications and MI within 180 days after initiation of NSAID (rofecoxib, diclofenac, ibuprofen, and naproxen compared with celecoxib) therapy was assessed using instrumental variable analysis. RESULTS: Compared with nonselective NSAIDs, celecoxib reduced the risk of GI complications by 1.4 per 100 users but increased the risk of MI by 0.3 per 100 users. Rofecoxib decreased GI complications by 1.1 per 100 users and increased the risk of MI by 0.3 per 100 users. Using celecoxib as the reference exposure showed an increase in the MI risk for rofecoxib (risk difference [RD] 1.40, 95% confidence interval [95% CI] 0.20, 3.01) and diclofenac (RD 6.07, 95% CI -0.02, 12.15). The RD for naproxen as well as its upper 95% CI was the lowest of all NSAIDs (RD -0.30, 95% CI -2.74, 2.14) and there was no significant difference in GI complication rates among all NSAIDs. CONCLUSION: In this instrumental variable analysis, diclofenac and rofecoxib had the least favorable benefit-risk balance among NSAIDs in older adults. PMID- 17075818 TI - HLA polymorphisms in African Americans with idiopathic inflammatory myopathy: allelic profiles distinguish patients with different clinical phenotypes and myositis autoantibodies. AB - OBJECTIVE: To investigate possible associations of HLA polymorphisms with idiopathic inflammatory myopathy (IIM) in African Americans, and to compare this with HLA associations in European American IIM patients with IIM. METHODS: Molecular genetic analyses of HLA-A, B, Cw, DRB1, and DQA1 polymorphisms were performed in a large population of African American patients with IIM (n = 262) in whom the major clinical and autoantibody subgroups were represented. These data were compared with similar information previously obtained from European American patients with IIM (n = 571). RESULTS: In contrast to European American patients with IIM, African American patients with IIM, in particular those with polymyositis, had no strong disease associations with HLA alleles of the 8.1 ancestral haplotype; however, African Americans with dermatomyositis or with anti Jo-1 autoantibodies shared the risk factor HLA-DRB1*0301 with European Americans. We detected novel HLA risk factors in African American patients with myositis overlap (DRB1*08) and in African American patients producing anti-signal recognition particle (DQA1*0102) and anti-Mi-2 autoantibodies (DRB1*0302). DRB1*0302 and the European American-, anti-Mi-2-associated risk factor DRB1*0701 were found to share a 4-amino-acid sequence motif, which was predicted by comparative homology analyses to have identical 3-dimensional orientations within the peptide-binding groove. CONCLUSION: These data demonstrate that North American IIM patients from different ethnic groups have both shared and distinct immunogenetic susceptibility factors, depending on the clinical phenotype. These findings, obtained from the largest cohort of North American minority patients with IIM studied to date, add additional support to the hypothesis that the myositis syndromes comprise multiple, distinct disease entities, perhaps arising from divergent pathogenic mechanisms and/or different gene-environment interactions. PMID- 17075819 TI - A novel autoantibody to a 155-kd protein is associated with dermatomyositis. AB - OBJECTIVE: In polymyositis and dermatomyositis (DM), identified autoantibodies occur in <50% of adult patients and in a smaller proportion of children. This study was undertaken as part of a larger effort to define novel autoantibodies that assist in the clinical evaluation of myositis. METHODS: Sera from children and adults satisfying criteria for idiopathic inflammatory myopathies and from patients with other connective tissue diseases (CTDs), patients with noninflammatory myopathies, and healthy individuals were tested for autoantibodies by immunoprecipitation (IP). A previously unrecognized autoantibody that immunoprecipitated a 155-kd protein along with a weaker 140-kd protein was seen. When the presence of this anti-p155 autoantibody in test sera was suggested based on IP results, it was confirmed by immunoblotting of immunoprecipitates. RESULTS: Sera from 51 of 244 myositis patients (21%), including 30 with juvenile DM (29%), 5 with juvenile CTD-associated myositis (33%), 8 with adult DM (21%), 6 with cancer-associated DM (75%), and 2 with adult CTD-associated myositis (15%), were found to have anti-p155 autoantibody. One of 49 patients with lupus, and none of 89 others without myositis, had anti-p155. Caucasian patients with anti-p155 had a unique HLA risk factor, DQA1*0301 (odds ratio 5.4, corrected P = 0.004). In adults with anti-p155, of several clinical features assessed only the frequency of V-sign rash was increased, but patients with this antibody were clinically distinct from those with autoantibodies to aminoacyl-transfer RNA synthetases. CONCLUSION: A newly recognized autoantibody, anti-p155, is associated with DM and cancer-associated DM, and is one of the most common autoantibodies in this condition, occurring as frequently in children as in adults. The clinical features and immunogenetics associated with anti-p155 differ from those associated with antisynthetases. PMID- 17075820 TI - Quantitative assessment of small intraosseous prostate cancer burden in SCID mice using fluorescence imaging. AB - BACKGROUND: Experimental bone metastases are typically analyzed when the skeletal tumor burden is large enough to be detected by imaging or histology. By this time, the bone microenvironment is usually destroyed, preventing useful analysis of tumor-bone interactions. METHODS: Small intraosseous tumors generated by intratibial injection of C4-2B prostate cancer cells transfected with green fluorescent protein (GFP) were assessed using in vivo and ex vivo fluorescence imaging, radiography, histology, and fluorometric analysis of bone lysates. RESULTS: Ex vivo fluorescence imaging and fluorometric analysis were capable of detecting tiny bone tumors as early as 10 days after injection. Ex vivo fluorescence imaging allowed simple quantification of small skeletal tumor burden and was useful in measuring the effect of systemic therapy. CONCLUSIONS: Ex vivo fluorescence imaging is a sensitive and easy method to quantify small skeletal tumor burden. This technique allows investigation of tumor-bone interactions while the bone microanatomy is still intact. PMID- 17075821 TI - Chronic bacterial infection and inflammation incite reactive hyperplasia in a mouse model of chronic prostatitis. AB - BACKGROUND: Chronic inflammation is postulated to contribute to prostate carcinogenesis. We developed a mouse model of chronic prostatitis to test whether infection-induced chronic inflammation would incite reactive changes in prostatic epithelium. METHODS: Prostate tissues harvested from either phosphate-buffered saline (PBS) or E. coli-infected mice were evaluated for histological changes and immunostained for markers of oxidative stress and epithelial cell proliferation. RESULTS: As compared to PBS-treated controls, mice infected with E. coli bacteria for 5 days showed foci of uniformly acute inflammation in the glandular lumen and a persistent inflammation at 12 weeks post-inoculation in the stroma. Prostatic glands showing varying degrees of atypical hyperplasia and dysplasia had stronger staining for oxidative DNA damage and increased epithelial cell proliferation than normal prostatic glands. CONCLUSIONS: These data demonstrate that chronic inflammation induces reactive hyperplasia associated with oxidative stress injury and support the proposed linkage among inflammation, oxidative DNA damage, and prostate carcinogenesis. PMID- 17075822 TI - Transfer of spermatogenesis-related cDNAs into eel testis germ-somatic cell coculture pellets by electroporation: methods for analysis of gene function. AB - Genes encoding spermatogenesis-related substance (eSRSs) show unique expression patterns during spermatogenesis. To analyze their function, we developed a new assay system using gene transfer techniques combined with coculture of the eel germ-somatic cells. First, we investigated the efficacy of in vitro electroporation transfer of gene into germ-somatic cell pellets using green fluorescent protein (GFP) gene. Second, in order to define the function of the eSRSs, we electrophoretically transferred eel spermatogonial stem cell renewal factor (eSRS34) and eel spermatogenesis-preventing substance (eSRS21) genes into germ-somatic cell pellets. Presence of the transferred cDNA was examined by reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, proliferating cells were detected histologically, after labeling with BrdU. Transfer of the eSRS34 gene induced spermatogonial stem cell renewal in the pellets. Moreover, 11-ketotestosterone (11-KT) treatment stimulated the proliferation of spermatogonia, which resulted in the appearance of late type B spermatogonia in the pellets. The proliferation of spermatogonia by 11-KT stimulation was suppressed by transfer of the eSRS21 gene. These results indicate that the transferred eSRS34 and 21genes were functional in the pellets. Thus, an efficient in vitro gene transfer technique for coculture system of germ and somatic cell of Japanese eel was established. PMID- 17075823 TI - Effectiveness of tumor necrosis factor inhibitors in rheumatoid arthritis in an observational cohort study: comparison of patients according to their eligibility for major randomized clinical trials. AB - OBJECTIVE: Randomized clinical trials (RCTs) evaluate the efficacy of treatments in selected groups of patients defined by strict inclusion criteria. The value of these trials in predicting therapeutic effectiveness in "real world" patients is limited. This observational cohort study was designed to complement the knowledge obtained in RCTs by evaluating the effectiveness of tumor necrosis factor (TNF) inhibitors in patients with rheumatoid arthritis (RA) according to their eligibility for the major trials. METHODS: Using the data from the German biologics register Rheumatoid Arthritis Observation of Biologic Therapy (RABBIT [in German]), we investigated how many of the RA patients who were treated with a TNF inhibitor (infliximab, etanercept, or adalimumab) would have been eligible for the major clinical trials that led to approval of the drugs. In addition, therapeutic effectiveness was compared in the eligible and ineligible patients using the American College of Rheumatology 20% (ACR20) and 50% (ACR50) improvement response criteria. RESULTS: Only 21-33% of the patients in the RABBIT register would have been eligible for the major trials. In these patients, the ACR20 and ACR50 improvement responses, indicating therapeutic effectiveness, were comparable with the response rates in the published trials. ACR response rates were lower in those patients considered ineligible for the trials; however, absolute improvement was similar to that in eligible patients. Ineligible patients had lower baseline disease activity, more comorbidity, and lower functional status. CONCLUSION: RCT cohorts reflect only a minor proportion of the patients treated with biologic agents in routine care. In the clinic setting, the indications for treatment with biologic agents are not identical to the inclusion criteria for trials. Despite the smaller relative improvement achieved in these patients with longstanding, severe RA who would not fulfill the inclusion criteria of a major trial, the majority of such patients would nevertheless benefit from biologic therapy. PMID- 17075824 TI - Estrogen signaling and disruption of androgen metabolism in acquired androgen independence during cadmium carcinogenesis in human prostate epithelial cells. AB - BACKGROUND: Lethal prostate cancers often become androgen-independent due to androgen receptor (AR) overexpression. The role of cadmium in prostate tumor progression was determined. METHODS: Control and cadmium-transformed prostate epithelial cells (CTPE) were compared for steroid-induced proliferation, steroid receptor expression, and androgen metabolism. RESULTS: CTPE cells showed rapid proliferation in complete medium and sustained proliferation in steroid-reduced medium. Androgens stimulated significantly less cell proliferation and AR-related genes expression in CTPE cells. 5alpha-Dihydrotestosterone increased PSA expression more effectively in control cells. Flutamide reduced 5alpha dihydrotestosterone-stimulated growth less effectively in CTPE cells compared to control. CTPE cells showed decreased p27 expression. Estrogen receptors were overexpressed and estradiol markedly stimulated proliferation in CTPE cells. In CTPE cells 5alpha-aromatase was markedly increased, while 5alpha-reductase was decreased. CONCLUSIONS: Cadmium-induced malignant transformation stimulates androgen independence, unrelated to AR expression or activity. Increased estrogen receptor and 5alpha-aromatase expression suggest estrogen signaling may be critical to this process. PMID- 17075825 TI - Increased serum levels of soluble fractalkine (CX3CL1) correlate with disease activity in rheumatoid vasculitis. AB - OBJECTIVE: To determine levels of soluble fractalkine (sFkn) in rheumatoid arthritis (RA) patients with and without rheumatoid vasculitis (RV), and to assess the relationship of sFkn levels to disease activity. METHODS: Serum was obtained from 98 RA patients (54 without vasculitis, 36 with extraarticular manifestations but without histologically proven vasculitis, and 8 with histologically proven vasculitis) and from 38 healthy individuals. Levels of sFkn were measured by enzyme-linked immunosorbent assay. Expression of Fkn and CX(3)CR1 was quantified by real-time polymerase chain reaction. Vasculitis disease activity was assessed using the Birmingham Vasculitis Activity Score and the Vasculitis Activity Index. RESULTS: Serum sFkn levels were significantly higher in patients with RA than in controls and were significantly higher in RA patients with RV than in those without vasculitic complications. Statistically significant correlations were observed between serum sFkn levels in RA patients and levels of C-reactive protein, rheumatoid factor, immune complex, and complement. In the RV group, sFkn levels also correlated with disease activity. Immunohistochemical analysis indicated that Fkn levels were associated mainly with endothelial cells in vasculitic arteries. In addition, expression of CX(3)CR1 messenger RNA was significantly greater in peripheral blood mononuclear cells from patients with active RV than in those from other RA patients or controls. Notably, serum sFkn levels were significantly diminished following successful treatment and clinical improvement. CONCLUSION: These findings suggest that Fkn and CX(3)CR1 play crucial roles in the pathogenesis of RV and that sFkn may serve as a serologic inflammatory marker of disease activity in RA patients with vasculitis. PMID- 17075827 TI - Clinical image: pigmented villonodular synovitis of the knee. PMID- 17075826 TI - Investigation of the MHC2TA gene, associated with rheumatoid arthritis in a Swedish population, in a UK rheumatoid arthritis cohort. AB - OBJECTIVE: A recent study of rheumatoid arthritis (RA) showed an association with a functional single-nucleotide polymorphism (SNP) mapping to the promoter region of the MHC2TA gene on chromosome 16p13 in a Swedish population. Interestingly, evidence for linkage to this region has been detected previously in a subgroup of UK RA families carrying 2 copies of shared epitope (SE) alleles. Therefore, we undertook this study to investigate the association of the MHC2TA gene promoter with RA in a UK Caucasian population. METHODS: Association with 5 SNPs spanning the promoter region of the MHC2TA gene was investigated in 813 UK RA patients and 532 population controls. Association with a functional putative RA-causal polymorphism (-168*G/A [rs3087456]) was tested in a total of 1,401 UK RA patients and 2,475 controls. Genotyping was performed using a Sequenom MassArray platform. Estimated haplotype frequencies were generated using the expectation-maximization algorithm and compared between patients and controls. RESULTS: All SNPs were in Hardy-Weinberg equilibrium. No evidence for association was found, either with the putative RA-causal polymorphism (-168*G/A) or with the other SNPs tested. Haplotype analysis revealed extensive linkage disequilibrium across the promoter region but no evidence for association. Stratifying the data set by carriage of SE alleles did not alter the conclusions. CONCLUSION: A functional polymorphism of the MHC2TA gene locus previously associated with RA in a European population has not been associated with RA in a UK population. These findings do not provide support for the notion that this gene plays a major role in the etiology of RA. PMID- 17075828 TI - A role for geranylgeranylation in interleukin-1beta secretion. AB - OBJECTIVE: Mevalonate kinase deficiency (MKD) is an autosomal-recessive disorder characterized by recurring episodes of inflammation. MK catalyzes the phosphorylation of mevalonic acid, which is an early step in isoprenoid biosynthesis. The goal of our study was to determine whether a temporary shortage of certain isoprenoid end products and/or the accumulation of mevalonic acid is the cause of interleukin-1beta (IL-1beta) secretion in MKD. METHODS: We studied the effect of the addition of intermediate metabolites and inhibitors of the isoprenoid biosynthesis pathway on IL-1beta secretion by peripheral blood mononuclear cells (PBMCs) of patients with MKD and healthy controls. RESULTS: Inhibition of enzymes involved in geranylgeranyl pyrophosphate (GGPP) synthesis or geranylgeranylation of proteins led to a marked increase of lipopolysaccharide stimulated IL-1beta secretion in PBMCs of control subjects. Furthermore, the increased IL-1beta secretion by PBMCs of patients with MKD was reversed by supplementation with GGPP as well as with mevalonic acid. IL-1beta secretion was increased only when control PBMCs were incubated with excessive amounts of mevalonic acid. Finally, a reduction in IL-1beta secretion by MKD PBMCs was also observed when sterol biosynthesis was inhibited, favoring nonsterol isoprenoid biosynthesis. CONCLUSION: Our results indicate that a shortage of geranylgeranylated proteins, rather than an excess of mevalonate, is likely to cause increased IL-1beta secretion by PBMCs of patients with MKD. PMID- 17075829 TI - Activation of nitric oxide signaling by the rheumatoid arthritis shared epitope. AB - OBJECTIVE: Susceptibility to rheumatoid arthritis (RA) is closely associated with HLA-DRB1 alleles encoding a shared epitope (SE) in positions 70-74 of the HLA DRbeta chain. The mechanistic basis for this association is unknown. Given the proposed pathogenic role of nitric oxide (NO) in RA, this study was undertaken to examine whether the SE can trigger NO signaling events. METHODS: The intracellular levels of NO were measured with the fluorescent NO probe 4,5 diaminofluorescein diacetate and by the 2,3-diaminonaphthalene method. NO synthase activity was determined by measuring the rate of conversion of radioactive arginine to citrulline. Levels of cGMP were measured with a commercial enzyme-linked immunosorbent assay, and the cytolytic activity of T cells was measured using a standard (51)Cr release assay. RESULTS: Lymphoblastoid B cell lines carrying SE-positive HLA-DR alleles displayed a higher rate of spontaneous NO production compared with SE-negative cells. L cell transfectants expressing SE-positive DR molecules on their surface also generated higher levels of NO. Tetrameric HLA-DR molecules containing a DRbeta-chain encoded by the SE positive DRB1*0401 allele stimulated fibroblast cells to produce higher levels of NO compared with cells stimulated with a control HLA-DR tetramer. Multimeric hepatitis B core proteins engineered to express region 65-79 encoded by the DRB1*0401 allele, but not the same region encoded by the control allele DRB1*0402, stimulated NO production in fibroblasts. Similarly, synthetic 15-mer peptides corresponding to the region 65-79 encoded by SE-positive alleles triggered increased NO levels when incubated with class II major histocompatibility complex-negative cells. The signaling pathway was found to involve NO synthase activation, followed by increased production of cGMP. SE triggered increased NO levels inhibited cytolytic elimination of target cells. CONCLUSION: The SE can trigger NO-mediated signaling events in opposite cells, and may thereby contribute to RA pathogenesis. PMID- 17075830 TI - Lethality in Drosophila melanogaster/Drosophila simulans species hybrids is not associated with substantial transcriptional misregulation. AB - The cross of Drosophila melanogaster females to Drosophila simulans males produces lethal F1 hybrid males. These lethal phenotypes can be suppressed by mutations in the D. melanogaster gene Hybrid male rescue (Hmr), demonstrating that Hmr has a major role in causing lethality in this hybridization. We performed parallel crosses to generate viable (Hmr-) and lethal (Hmr+) hybrid male larvae and used microarrays to compare whole-genome transcriptional profiles between these two samples. This comparison was done to investigate two questions: whether hybrid lethality is associated with substantial gene misregulation, and whether a mechanistic basis for hybrid lethality can be inferred from the identities of differentially expressed individual transcripts. We report that a surprisingly small number of genes have a significant difference in transcript abundance between lethal and viable hybrid males. There is a significant over representation of genes encoding proteosome subunits among those upregulated in lethal hybrids relative to viable hybrids. Genetic tests, however, failed to fully support the hypothesis that this overexpression is causing hybrid lethality. Hybrid females were previously reported to have a significantly different expression pattern of sex-biased genes compared to the parental species. We find no such differences between lethal and viable hybrid males. We did find a significant deficit of X chromosome genes among those downregulated in lethal hybrids, but not among those upregulated. We suggest that while interspecific hybrids may have substantial amounts of gene misregulation compared to their parental species, many of these transcriptional differences may be only indirectly related to hybrid incompatibility phenotypes. PMID- 17075831 TI - Aberrant expression of SWI/SNF catalytic subunits BRG1/BRM is associated with tumor development and increased invasiveness in prostate cancers. AB - BACKGROUND: Brahma gene (BRM) and Brahma-related gene 1 (BRG1) are major components with ATPase enzymatic activities in the nucleosome remodeling SWI/SNF complex, and their expression pattern in human prostate cancers is unknown. METHOD: We analyzed a published cDNA microarray data set of prostate cancers for the expression of SWI/SNF genes, and then we evaluated the expression levels of BRG1 and BRM proteins with a semi-quantitative immunohistochemistry (IHC) approach in a pairwise manner of malignant versus benign tissues from individual prostate cancers. The correlation of BRG1/BRM expression with clinical parameters was analyzed. RESULTS: Microarray data showed an aberrant expression of BRG1 and BRM but not SNF5/INI1 genes in different stages of the disease course. In immunochemistry studies, BRG1 expression was significantly higher in malignant tissues compared to their benign compartments, and this difference was more profound in high-grade cancers. Although BRM expression showed a heterogeneous pattern, the average level of BRM expression was lower in malignant tissues than that in benign tissues. More interestingly, BRG1 and BRM expression showed a reciprocal pattern in both benign and malignant tissues of individual cases. In malignant tissues, higher BRG1 but not BRM expression levels were associated with larger volume of tumor mass. Increased expression of BRG1 but not BRM protein was observed in invasive cancer cells. Consistently, overexpression of exogenous wild type BRG1 and BRM but not mutant BRG1 enhanced cancer cell invasion in an in vitro cell invasion assay. CONCLUSIONS: We provide the first evidence that aberrant expression of BRG1 and BRM genes is associated with disease development and progression in prostate cancers and increased BRG1 expression may promote tumor growth and invasion. PMID- 17075832 TI - Cytoskeletal and nuclear organization in mouse embryos derived from nuclear transfer and ICSI: a comparison of agamogony and syngamy before and during the first cell cycle. AB - In this study, somatic cell nuclear transfer (SCNT) and intracytoplasmic sperm injection (ICSI) are used as models of agamogony and syngamy, respectively. In order to elucidate the reasons of low efficiency of somatic cell cloning, cytoskeletal and nuclear organization in cloned mouse embryos was monitored before and during the first cell cycle, and compared with the pattern of ICSI zygote. A metaphase-like spindle with alignment of condensed donor chromosomes was assembled within 3 hr after NT, followed by formation of pronuclear-like structures at 3-6 hr after activation, indicating that somatic nuclear remodeling depends on microtubular network organization. The percentage of two (pseudo-) pronuclei in cloned embryos derived from delayed activation was greater than that in immediate activation group (68.5% vs. 30.8%, P<0.01), but similar to that of ICSI group (68.5% vs. 65.5%, P>0.05). The 2-cell rate in NT embryos was significantly lower than that in zygotes produced by ICSI (64.8% vs. 82.5%, P<0.01). Further studies testified that the cloned embryos reached the metaphase of the first mitosis 10 hr after activation, whereas this occurred at 18 hr in the ICSI zygotes. Comparision of the pattern of microfilament assembly in early NT embryos with that in syngamic zygotes suggested that abnormal microfilamental pattern in cloned embryos may threaten subsequent embryonic development. In conclusion, agamogony, in contrast to syngamy, displays some unique features in respect of cytoskeletal organization, the most remarkable of which is that the first cell cycle is initiated ahead distinctly, which probably leads to incomplete organization of the first mitotic spindle, and contributes to low efficiency of cloning. PMID- 17075833 TI - Survivin acts as anti-apoptotic factor during the development of bovine pre implantation embryos. AB - Survivin, an inhibitor of apoptotic protein containing a single baculoviral inhibit apoptotic protein repeat domain, is a bifunctional protein that suppresses apoptosis and regulates cell division. Thus, we used double stranded RNA (dsRNA) interference to manipulate survivin expression in bovine embryos and analyze its role in blocking apoptosis and facilitating development of pre implantation embryos. In vitro fertilized embryos (1-cell) were injected with survivin dsRNA, and expression of survivin mRNA was evaluated by real-time quantitative RT-PCR. To analyze survivin protein expression, we performed immunocytochemistry using a rabbit anti-bovine suvivin antibody. Expression levels of survivin mRNA and protein were decreased in the dsRNA group compared to the sham group. Rates of in vitro blastocyst development were lower in the survivin dsRNA-injected group than in the sham-injected group. Also, the total cell number seen in blastocysts was decreased in the dsRNA group. TUNEL assays of DNA fragmentation indicated an increased apoptotic index in the dsRNA group compared to the sham group. These results indicate that survivin is important for optimal development of bovine blastocysts and confirm that survivin expression suppresses apoptosis of pre-implantation embryos. PMID- 17075834 TI - Cell cycle analysis and interspecies nuclear transfer of in vitro cultured skin fibroblasts of the Siberian tiger (Panthera tigris Altaica). AB - The present study was conducted to examine the effect of cell culture conditions, antioxidants, protease inhibitors (PI), and different levels of dimethylsulfoxide (DMSO) for the promotion of synchronization of different cell cycles of Siberian tiger skin fibroblasts. We also compared the ability of somatic cell nuclei of the Siberian tiger in pig cytoplasts and to support early development after reconstruction. Cell cycle synchronization between nuclear donor and recipient cells is considered to be one of the most crucial factors for successful cloning. Five experiments were performed each with a one-way completely randomized design involving three replicates of all treatments. Least significant difference (LSD) was used to determine variation among treatment groups. Experiment I focused in the effects of cycling, serum starved and fully confluent stages of Siberian tiger cells on different cell cycles. In Experiment II, the effects of different antioxidants like beta-Mercaptoethanol (beta-ME, 10 microM), cysteine (2 mM), and glutathione (2 mM) were examined after cells were fully confluent without serum starvation for 4 hr. In Experiment III, three PI, namely 6-dimethylaminopurine (6 DMAP, 2 mM), cycloheximide (7.5 microg/ml) and cytochalasin B (7.5 microg/ml) were used in the sane manner as in Experiment II. In Experiment IV, different levels of DMSO at 0%, 0.5%, 1.0%, and 2.5% were tested on different cell cycle stages of Siberian tiger examined by Flowcytometry (FACS). In Experiment I, 67.2% of the Siberian tiger skin fibroblasts reached the G0/G1 stage (2C DNA content) in fully confluent conditions which was more than the cycling (49.8%) and serum starved (SS) medium (65.5%; P < 0.05). Among the chemically treated group, glutathione (72.6%) and cycloheximide (71.3%) had little bit better results for the synchronization of G0 + G1 phases than serum starved and fully confluent. After nuclear transfer we did not see any significant differences on the development of tiger-porcine reconstructed embryos at cycling, SS and fully confluent. Data indicate that prolonged culture of cells in the absence of serum as well as using different chemicals for this experiment does not imply a shift in the percentage of cells that enter G0/G1 and that confluency is sufficient to induce quiescence. This finding can be beneficial in nuclear transfer programs in Siberian tiger, because there are negative effects, such as apoptosis associated with serum starvation. PMID- 17075835 TI - Glycan analysis of monoclonal antibodies secreted in deposition disorders indicates that subsets of plasma cells differentially process IgG glycans. AB - OBJECTIVE: To compare the glycosylation of polyclonal serum IgG heavy chains in a patient with rheumatoid arthritis (RA) with that of monoclonal serum IgG heavy chains in the same patient during an episode of heavy-chain deposition disease (HCDD), to establish whether glycosylation processing is specific for subsets of B cells. METHODS: Serum IgG was purified using a HiTrap protein G column. Immunoglobulins were run on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels, and IgG glycans were isolated from gel bands and fluorescently labeled. Glycans were analyzed by normal-phase high-performance liquid chromatography and by liquid chromatography-electrospray ionization-mass spectrometry. RESULTS: The glycosylation of serum immunoglobulins from a patient with seronegative RA and HCDD was analyzed. The predominant immunoglobulin was a truncated glycosylated gamma3 heavy chain, and a small amount of polyclonal IgG was also present. The glycan profile showed that the monoclonal gamma3 heavy chain contained fully galactosylated biantennary glycans with significantly less fucose but more sialic acid than in IgG3 from healthy controls. In contrast, the polyclonal IgG showed an RA-like profile, with a predominance of fucosylated biantennary glycans and low levels of galactosylation. The glycan profile of serum IgG obtained from the same patient during disease remission resembled a typical RA profile. CONCLUSION: These data indicate that different types of B cells process a particular set of IgG glycoforms. PMID- 17075836 TI - RhoA-mediated, tumor necrosis factor alpha-induced activation of NF-kappaB in rheumatoid synoviocytes: inhibitory effect of simvastatin. AB - OBJECTIVE: Increasing evidence indicates that RhoA may play a central role in the inflammatory response. This study was conducted to examine the role of RhoA in mediating the activation of NF-kappaB in tumor necrosis factor alpha (TNFalpha) stimulated rheumatoid synoviocytes, and to evaluate the modulatory effects of statins on the TNFalpha-induced activation of RhoA and NF-kappaB and the secretion of proinflammatory cytokines by rheumatoid synoviocytes. METHODS: Rheumatoid synoviocytes obtained from patients with active rheumatoid arthritis were stimulated with TNFalpha and incubated with simvastatin (SMV) (1 muM). RhoA activity was assessed by a pull-down assay. NF-kappaB DNA binding activity and nuclear translocation of NF-kappaB were measured by a sensitive multiwell colorimetric assay and confocal fluorescence microscopy, respectively. RESULTS: TNFalpha stimulation elicited a robust increase in RhoA activity in a dose dependent manner, and SMV mitigated this increase. TNFalpha also hastened NF kappaB nuclear translocation of subunit p65 and increased DNA binding activity, luciferase reporter gene expression, degradation of IkappaB, and secretion of interleukin-1beta (IL-1beta) and IL-6. SMV prevented the increase in NF-kappaB activation and rise in IL-1beta and IL-6 levels induced by TNFalpha, whereas mevalonate and geranylgeranyl pyrophosphate reversed the inhibitory effects of SMV on activation of NF-kappaB and RhoA. Furthermore, cotransfection with a dominant-negative mutant of RhoA demonstrated that the TNFalpha-induced signaling pathway involved sequential activation of RhoA, leading to NF-kappaB activation and, ultimately, to secretion of cytokines. CONCLUSION: This study identifies RhoA as the key regulator of TNFalpha-induced NF-kappaB activation, which ultimately results in the secretion of proinflammatory cytokines in rheumatoid synoviocytes. The findings provide a new rationale for the antiinflammatory effects of statins in inflammatory arthritis. PMID- 17075837 TI - The scapuloacromial angle: a 3D analysis of acromial slope and its relationship with shoulder impingement. AB - PURPOSE: To quantify acromial slope (the scapuloacromial [SA] angle). MATERIALS AND METHODS: We used shoulder MR studies in 64 subjects who were stratified surgically into impingement (with and without tear) and nonimpingement (shoulder instability) groups. We measure, in three dimensions, the angle between unit normal vectors to planes of best fit to the undersurface of the acromion and the medial surface of the blade of the scapula. We examine the projection of this SA angle onto the transverse, coronal, and sagittal planes to gauge some measure of rotation, lateral tilt, and anterior tilt with respect to the patient. Orientation of the acromion and scapular blade in relation to the MR scanner and therefore to the patient was also calculated. RESULTS: We found no significant differences between these two groups when examining age and side of pathology. A statistically significant difference was seen between males and females. CONCLUSION: Our study does not support the idea that a flatter acromial slope causes impingement and is an etiological basis for rotator cuff tears. We believe that acromial slope, regardless of imaging plane, is not useful in classifying patients who present with shoulder pain and should not be considered as a source of pathologic changes. PMID- 17075838 TI - Elevated mean diffusivity in widespread brain regions in congenital central hypoventilation syndrome. AB - PURPOSE: To investigate whether mean diffusivity (MD) values are altered in brain areas underlying cardiovascular and respiratory control in congenital central hypoventilation syndrome (CCHS). MATERIALS AND METHODS: Conventional and diffusion tensor imaging were performed in 15 CCHS and 30 control subjects, using a 3.0-Tesla MRI unit. Mean diffusivity maps were calculated from diffusion weighted images, spatially normalized, smoothed, and compared between groups using analysis of covariance at each voxel with age as covariate. Global mean MD values of gray and white matter were determined in individual subjects and compared between groups and with age. RESULTS: Increased MD values appeared in CCHS over control subjects within multiple areas influencing breathing and cardiovascular control, including the midbrain, pons, and dorsal and ventral medulla. Other altered sites included cerebellar cortex and deep nuclei, basal ganglia, basal forebrain, and temporal and frontal cortices. Global mean MD values for gray and white matter did not differ between groups; however, gray matter MD values significantly increased with age (P < 0.02) in CCHS patients only. CONCLUSION: Increased MD values suggest regional alterations or injury; affected areas include brainstem sites classically associated with autonomic and respiratory control. Other altered regions mediate additional physiological characteristics impaired in CCHS. PMID- 17075839 TI - Determination of the hydrolysis kinetics of alpha-naphthyl acetate in micellar systems and the effect of HPMC (catalyst present). AB - The change in the hexadecyltrimethylammonium bromide (CTAB) critical aggregation concentration (CAC) was studied in the presence of various concentrations and grades of hydroxypropylmethyl cellulose (HPMC) using surface tension measurement (duNouy ring and Wilhelmy plate) and oil red O solubilization. According to the surface tension methods, the CAC was higher than the CTAB critical micelle concentration (CMC). CAC and CMC were not different when the solubilization method was used. Micellar solutions of CTAB have been found to accelerate the hydrolysis of alpha-naphthyl acetate (alpha-NA) by o-iodosobenzoic acid (IBA), a strong nucleophile. Pseudo-first-order kinetics were utilized for rate constant determination. The observed rate constants for the degradation of alpha-NA in the presence of varying CTAB concentrations with and without HPMC were analyzed according to the pseudophase model. The micellar rate constants and the micellar binding constants for the substrates were obtained. The presence of HPMC retarded the reaction rate, and the rate constant decreased as the polymer concentration increased. However, there was no obvious difference in the observed rate constants among the different grades of HPMC (Methocel E5, Methocel E15, Methocel E50). The decrease in the rate constant was likely due to the polymer-micelle interaction interfering with substrate binding to the CTAB micelles. PMID- 17075840 TI - Efficacy and mechanism of action of turmeric supplements in the treatment of experimental arthritis. AB - OBJECTIVE: Scientific evidence is lacking for the antiarthritic efficacy of turmeric dietary supplements that are being promoted for arthritis treatment. Therefore, we undertook studies to determine the antiarthritic efficacy and mechanism of action of a well-characterized turmeric extract using an animal model of rheumatoid arthritis (RA). METHODS: The composition of commercial turmeric dietary supplements was determined by high-performance liquid chromatography. A curcuminoid-containing turmeric extract similar in composition to these supplements was isolated and administered intraperitoneally to female Lewis rats prior to or after the onset of streptococcal cell wall-induced arthritis. Efficacy in preventing joint swelling and destruction was determined clinically, histologically, and by measurement of bone mineral density. Mechanism of action was elucidated by analysis of turmeric's effect on articular transcription factor activation, microarray analysis of articular gene expression, and verification of the physiologic effects of alterations in gene expression. RESULTS: A turmeric fraction depleted of essential oils profoundly inhibited joint inflammation and periarticular joint destruction in a dose dependent manner. In vivo treatment prevented local activation of NF-kappaB and the subsequent expression of NF-kappaB-regulated genes mediating joint inflammation and destruction, including chemokines, cyclooxygenase 2, and RANKL. Consistent with these findings, inflammatory cell influx, joint levels of prostaglandin E(2), and periarticular osteoclast formation were inhibited by turmeric extract treatment. CONCLUSION: These translational studies demonstrate in vivo efficacy and identify a mechanism of action for a well-characterized turmeric extract that supports further clinical evaluation of turmeric dietary supplements in the treatment of RA. PMID- 17075841 TI - Methods and applications of diffusion imaging of vertebral bone marrow. AB - Diffusion-weighted imaging (DWI) is an MRI technique that is sensitive to random water movements at spatial scales far below typical MRI voxel dimensions. DWI is a valuable tool for the diagnoses of diseases that involve alterations in water mobility. In the spine, DWI has proven to be a highly useful method for the differential diagnosis of benign and malignant compression fractures. In these pathologies, the microscopic structure of bone marrow is altered in a very different ways, leading to different water mobility, which can be depicted by DWI. Most of the pulse sequences developed for MRI can be adapted for DWI. However, these DWI-adapted sequences are frequently affected by artifacts, mostly caused by physiological motion. Therefore, the introduction of additional correction techniques, or even the development of new sequences is necessary. The first part of this article describes the principles of DWI and the sequences used for DWI of the spine: spin echo (SE), turbo spin echo (TSE), single-shot echo planar imaging (EPI), and steady-state free precession (SSFP) sequences. In the second part, clinical applications of DWI of the spinal bone marrow are extensively discussed. PMID- 17075842 TI - Valproic acid-induced fetal malformations are reduced by maternal immune stimulation with granulocyte-macrophage colony-stimulating factor or interferon gamma. AB - Valproic acid, a drug commonly used to treat seizures and other psychiatric disorders, causes neural tube defects (NTDs) in exposed fetuses at a rate 20 times higher than in the general population. Failure of the neural tube to close during development results in exencephaly or anencephaly, as well as spina bifida. In mice, nonspecific activation of the maternal immune system can reduce fetal abnormalities caused by diverse etiologies, including diabetes-induced NTDs. We hypothesized that nonspecific activation of the maternal immune system with interferon-gamma (IFN-gamma) and granulocyte-macrophage colony-stimulating factor (GM-CSF) could reduce valproic acid (VA)-induced defects as well. Female CD-1 mice were given immune stimulant prebreeding: either IFN-gamma or GM-CSF. Approximately half of the control and immune-stimulated pregnant females were then exposed to 500 mg/kg VA on the morning of gestational day 8. The incidence of developmental defects was determined on gestational day 17 from at least eight litters in each of the following treatment groups: control, VA only, IFN-gamma only, IFN-gamma+VA, GM-CSF only, and GM-CSF+VA. The incidence of NTDs was 18% in fetuses exposed to VA alone, compared to 3.7% and 2.9% in fetuses exposed to IFN gamma+VA, or GM-CSF+VA respectively. Ocular defects were also significantly reduced from 28.0% in VA exposed groups to 9.8% in IFN-gamma+VA and 12.5% in GM CSF+VA groups. The mechanisms by which maternal immune stimulation prevents birth defects remain unclear, but may involve maternal or fetal production of cytokines or growth factors which protect the fetus from the dysregulatory effects of teratogens. PMID- 17075843 TI - Involvement of specific laminins and nidogens in the active remodeling of the basal lamina of labial salivary glands from patients with Sjogren's syndrome. AB - OBJECTIVE: To investigate remodeling of the basal lamina of labial salivary glands (LSGs) from patients with Sjogren's syndrome (SS) by analyzing the expression of specific components that participate in its assembly and attachment to acinar and ductal cells. METHODS: Two groups of SS patients with similar levels of remnant glandular tissue but with low and high levels of interacinar fibrosis, respectively, were studied. The expression of laminin alpha1, alpha4, and gamma2 chains and nidogens was examined at the messenger RNA (mRNA) and protein levels. Nidogens 1 and 2 were also studied in situ by immunohistochemistry. RESULTS: Increases in the amount of mRNA and protein for both the processed and unprocessed laminin gamma2-chain were more pronounced in patients with low interacinar fibrosis. Increases in the protein levels of laminin alpha1 and alpha4 chains were observed in patients with low interacinar fibrosis, but not in those with high interacinar fibrosis. Nidogen mRNA and protein levels were similar in SS patients and controls. Interestingly, high levels of nidogen degradation were observed in patients with low interacinar fibrosis. Nidogens were readily detected by immunofluorescence in the basal lamina of the capillaries and stroma in SS patients, but were less apparent in the basal lamina of the acini and ducts. CONCLUSION: These results suggest that the basal lamina of LSGs from patients with SS is undergoing active remodeling, such that alterations are less evident in patients who have advanced morphologic signs of the disease (high interacinar fibrosis). Nidogen proteolysis might account for the disorganization of the basal lamina that is typically observed in LSGs from SS patients, assuming that cleavage impairs their ability to crosslink type IV collagen and laminin networks. PMID- 17075844 TI - Human cranial anatomy and the differential preservation of population history and climate signatures. AB - Cranial morphology is widely used to reconstruct evolutionary relationships, but its reliability in reflecting phylogeny and population history has been questioned. Some cranial regions, particularly the face and neurocranium, are believed to be influenced by the environment and prone to convergence. Others, such as the temporal bone, are thought to reflect more accurately phylogenetic relationships. Direct testing of these hypotheses was not possible until the advent of large genetic data sets. The few relevant studies in human populations have had intriguing but possibly conflicting results, probably partly due to methodological differences and to the small numbers of populations used. Here we use three-dimensional (3D) geometric morphometrics methods to test explicitly the ability of cranial shape, size, and relative position/orientation of cranial regions to track population history and climate. Morphological distances among 13 recent human populations were calculated from four 3D landmark data sets, respectively reflecting facial, neurocranial, and temporal bone shape; shape and relative position; overall cranial shape; and centroid sizes. These distances were compared to neutral genetic and climatic distances among the same, or closely matched, populations. Results indicate that neurocranial and temporal bone shape track neutral genetic distances, while facial shape reflects climate; centroid size shows a weak association with climatic variables; and relative position/orientation of cranial regions does not appear correlated with any of these factors. Because different cranial regions preserve population history and climate signatures differentially, caution is suggested when using cranial anatomy for phylogenetic reconstruction. PMID- 17075845 TI - Overexpression of phosphorylated STAT-1alpha in the labial salivary glands of patients with Sjogren's syndrome. AB - OBJECTIVE: To clarify the molecular mechanisms of Sjogren's syndrome (SS), we analyzed the functional role of the STAT-1 gene, one of the interferon-gamma (IFNgamma)-inducible genes, in labial salivary glands (LSGs) from SS patients. METHODS: The expression of STAT-1 messenger RNA (mRNA) was examined by real-time polymerase chain reaction (PCR) analysis, and the phosphorylation of STAT-1 protein (Tyr(701) and Ser(727) pSTAT-1) was investigated by Western blot and immunohistochemical analyses. The expression of IFNgamma-inducible 10-kd protein (IP-10), IFN regulatory factor 1 (IRF-1), and Fas was also examined by real-time PCR and immunohistochemical analyses. RESULTS: STAT-1alpha and STAT-1beta mRNA were highly expressed in LSGs from SS patients. The level of STAT-1alpha protein in SS LSGs was higher than that in 3 control LSGs, whereas STAT-1beta protein was not clearly detected by Western blot analysis. Moreover, Tyr(701) and Ser(727) pSTAT-1alpha proteins were specifically detected in SS LSGs. Immunohistochemical analysis showed localization of Tyr(701) pSTAT-1 in infiltrating lymphocytes and the adjacent ductal epithelium from SS patients. Ser(727) pSTAT-1 was localized only in the ductal epithelium of SS LSGs. The STAT-1-inducible genes IP-10 and IRF-1 and the Fas genes were highly expressed in SS LSGs and were colocalized with Ser(727) pSTAT-1-positive, but not Tyr(701) pSTAT-1-positive, cells. CONCLUSION: We found evidence of the up-regulation of STAT-1alpha mRNA and protein in LSGs from SS patients, as well as the presence of pSTAT-1alpha in ductal epithelium from SS patients. Our findings suggest that STAT-1alpha, especially Ser(727) pSTAT-1, may function as a key molecule in the pathogenesis of SS. PMID- 17075846 TI - Remodeling dynamics in the alveolar process in skeletally mature dogs. AB - Bone turnover rates can be altered by metabolic and mechanical demands. Due to the difference in the pattern of loading, we hypothesized that there are differences in bone remodeling rates between the maxillary and mandibular alveolar processes. Furthermore, in a canine model, the alveolar process of teeth that lack contact (e.g., second premolars) would have a different turnover rate than bone supporting teeth with functional contact (e.g., first molars). Six skeletally mature male dogs were given a pair of calcein labels. After sacrifice, specimens representing the anterior and posterior locations of both jaws were prepared for examination by histomorphometric methods to evaluate the bone volume/total volume (BV/TV; %), bone volume (mm2), mineral apposition rate (MAR; microm/day), and bone formation rate (BFR; %/year) in the alveolar process. There were no significant differences (P>0.05) in the BV/TV within the jaws. The bone volume within the alveolar process of the mandible was 2.8-fold greater than in the maxilla. The MAR was not significantly different between the jaws and anteroposterior locations. However, the BFR was significantly (P<0.0001) greater in the mandible than in the maxilla. The anterior location had higher (P=0.002) remodeling than the posterior location in the maxilla but not in the mandible. While there was a greater bone mass and increased remodeling in the mandible, no remodeling gradient in the coronal-apical direction was apparent in the alveolar process. Bone adaptation probably involves a complex interplay of bone turnover, mass, and architecture. PMID- 17075847 TI - Epicardium-derived cells are important for correct development of the Purkinje fibers in the avian heart. AB - During embryonic development, the proepicardial organ (PEO) grows out over the heart surface to form the epicardium. Following epithelial-mesenchymal transformation, epicardium-derived cells (EPDCs) migrate into the heart and contribute to the developing coronary arteries, to the valves, and to the myocardium. The peripheral Purkinje fiber network develops from differentiating cardiomyocytes in the ventricular myocardium. Intrigued by the close spatial relationship between the final destinations of migrating EPDCs and Purkinje fiber differentiation in the avian heart, that is, surrounding the coronary arteries and at subendocardial sites, we investigated whether inhibition of epicardial outgrowth would disturb cardiomyocyte differentiation into Purkinje fibers. To this end, epicardial development was inhibited mechanically with a membrane, or genetically, by suppressing epicardial epithelial-to-mesenchymal transformation with antisense retroviral vectors affecting Ets transcription factor levels (n=4, HH39-41). In both epicardial inhibition models, we evaluated Purkinje fiber development by EAP-300 immunohistochemistry and found that restraints on EPDC development resulted in morphologically aberrant differentiation of Purkinje fibers. Purkinje fiber hypoplasia was observed both periarterially and at subendocardial positions. Furthermore, the cells were morphologically abnormal and not aligned in orderly Purkinje fibers. We conclude that EPDCs are instrumental in Purkinje fiber differentiation, and we hypothesize that they cooperate directly with endothelial and endocardial cells in the development of the peripheral conduction system. PMID- 17075848 TI - Pulmonary intravascular monocytes/macrophages in a rat model of sepsis. AB - Sepsis induces recruitment of neutrophils and monocytes/macrophages in the lung and enhances host susceptibility to a secondary bacterial challenge. The phenotype and functions of recruited pulmonary intravascular monocytes/macrophages (PIMMs) in sepsis remain largely unknown. Therefore, we characterized PIMM recruitment and functions in a rat model of E. coli-induced sepsis. Male Sprague-Dawley rats were injected intraperitoneally with saline (n=10) and 48 hr after the saline treatment treated intravenously with either saline (n=5) or E. coli lipopolysachharide (LPS; 1.5 microg/kg body weight; n=5). A second group of 10 rats was infected intraperitoneally with E. coli (2x10(7) CFU/100 g) followed by intravenous injection of either saline (n=5) or LPS (n=5) 48 hr after the first treatment. Rats were euthanized at 6 hr after LPS treatment. Immunocytochemistry showed more PIMMs stained with ED-1 antibody, which specifically reacts with rat monocytes/macrophages, in rats infected with E. coli compared with the controls (P<0.05). LPS treatment of E. coli-infected rats increased the numbers of PIMMs (P<0.05) and induced more inflammation compared to other groups. Immuno-electron microscopy localized TNF-alpha, IL-10, and TGF-beta2 in recruited PIMMs in rats challenged with both E. coli and LPS. ELISA on lung homogenates showed higher concentrations of TNF-alpha, IL-10, and TGF-beta2 in rats treated with both E. coli and LPS compared with those treated with only LPS or E. coli (P<0.05). We conclude that ED-1-positive PIMMs are recruited in this model of sepsis and contain TNF-alpha, IL-10, and TGF-beta2. PMID- 17075849 TI - Acupuncture in patients with osteoarthritis of the knee or hip: a randomized, controlled trial with an additional nonrandomized arm. AB - OBJECTIVE: To investigate the effectiveness of acupuncture in addition to routine care, compared with routine care alone, in the treatment of patients with chronic pain due to osteoarthritis (OA) of the knee or hip. METHODS: In a randomized, controlled trial, patients with chronic pain due to OA of the knee or hip were randomly allocated to undergo up to 15 sessions of acupuncture in a 3-month period or to a control group receiving no acupuncture. Another group of patients who did not consent to randomization underwent acupuncture treatment. All patients were allowed to receive usual medical care in addition to the study treatment. Clinical OA severity (Western Ontario and McMaster Universities Osteoarthritis Index [WOMAC]) and health-related quality of life (Short Form 36) were assessed at baseline and after 3 months and 6 months. RESULTS: Of 3,633 patients (mean +/- SD age 61.8 +/- 10.8 years; 61% female), 357 were randomized to the acupuncture group and 355 to the control group, and 2,921 were included in the nonrandomized acupuncture group. At 3 months, the WOMAC had improved by a mean +/- SEM of 17.6 +/- 1.0 in the acupuncture group and 0.9 +/- 1.0 in the control group (3-month scores 30.5 +/- 1.0 and 47.3 +/- 1.0, respectively [difference in improvement 16.7 +/- 1.4; P < 0.001]). Similarly, quality of life improvements were more pronounced in the acupuncture group versus the control group (P < 0.001). Treatment success was maintained through 6 months. The changes in outcome in nonrandomized patients were comparable with those in randomized patients who received acupuncture. CONCLUSION: These results indicate that acupuncture plus routine care is associated with marked clinical improvement in patients with chronic OA-associated pain of the knee or hip. PMID- 17075850 TI - Analysis of coating structures and interfaces in solid oral dosage forms by three dimensional terahertz pulsed imaging. AB - Three dimensional terahertz pulsed imaging (TPI) was evaluated as a novel tool for the nondestructive characterization of different solid oral dosage forms. The time-domain reflection signal of coherent pulsed light in the far infrared was used to investigate film-coated tablets, sugar-coated tablets, multilayered controlled release tablets, and soft gelatin capsules. It is possible to determine the spatial and statistical distribution of coating thickness in single and multiple coated products using 3D TPI. The measurements are nondestructive even for layers buried underneath other coating structures. The internal structure of coating materials can be analyzed. As the terahertz signal penetrates up to 3 mm into the dosage form interfaces between layers in multilayered tablets can be investigated. In soft gelatin capsules it is possible to measure the thickness of the gelatin layer and to characterize the seal between the gelatin layers for quality control. TPI is a unique approach for the nondestructive characterization and quality control of solid dosage forms. The measurements are fast and fully automated with the potential for much wider application of the technique in the process analytical technology scheme. PMID- 17075851 TI - Risedronate decreases biochemical markers of cartilage degradation but does not decrease symptoms or slow radiographic progression in patients with medial compartment osteoarthritis of the knee: results of the two-year multinational knee osteoarthritis structural arthritis study. AB - OBJECTIVE: Bisphosphonates have slowed the progression of osteoarthritis (OA) in animal models and have decreased pain in states of high bone turnover. The Knee OA Structural Arthritis (KOSTAR) study, which is the largest study to date investigating a potential structure-modifying OA drug, tested the efficacy of risedronate in providing symptom relief and slowing disease progression in patients with knee OA. METHODS: The study group comprised 2,483 patients with medial compartment knee OA and 2-4 mm of joint space width (JSW), as determined using fluoroscopically positioned, semiflexed-view radiography. Patients were enrolled in 2 parallel 2-year studies in North America and the European Union. These studies evaluated the efficacy of risedronate at dosages of 5 mg/day, 15 mg/day, 35 mg/week (in Europe), and 50 mg/week (in North America) compared with placebo in reducing signs and symptoms, as measured by the Western Ontario and McMaster Universities Osteoarthritis (WOMAC) index and patient global assessment (PGA) scores, and in slowing radiographic progression. RESULTS: A reduction of approximately 20% in signs and symptoms, as measured by WOMAC subscales and PGA scores, was observed in all groups, with no treatment effect of risedronate demonstrated. Risedronate did not significantly reduce radiographic progression as measured by decreased JSW or using a dichotomous definition of progression (joint space loss of >or=0.6 mm). Thirteen percent of patients receiving placebo demonstrated significant disease progression over 2 years. A dose-dependent reduction in the level of C-terminal crosslinking telopeptide of type II collagen, a cartilage degradation marker associated with progressive OA, was seen in patients who received risedronate. No increase in the number of adverse events was demonstrated for risedronate compared with placebo. CONCLUSION: Although risedronate (compared with placebo) did not improve signs or symptoms of OA, nor did it alter progression of OA, a reduction in the level of a marker of cartilage degradation was observed. A sustained clinically relevant improvement in signs and symptoms was observed in all treatment and placebo groups. PMID- 17075852 TI - 9.4T human MRI: preliminary results. AB - This work reports the preliminary results of the first human images at the new high-field benchmark of 9.4T. A 65-cm-diameter bore magnet was used together with an asymmetric 40-cm-diameter head gradient and shim set. A multichannel transmission line (transverse electromagnetic (TEM)) head coil was driven by a programmable parallel transceiver to control the relative phase and magnitude of each channel independently. These new RF field control methods facilitated compensation for RF artifacts attributed to destructive interference patterns, in order to achieve homogeneous 9.4T head images or localize anatomic targets. Prior to FDA investigational device exemptions (IDEs) and internal review board (IRB) approved human studies, preliminary RF safety studies were performed on porcine models. These data are reported together with exit interview results from the first 44 human volunteers. Although several points for improvement are discussed, the preliminary results demonstrate the feasibility of safe and successful human imaging at 9.4T. PMID- 17075853 TI - Lower prevalence of chondrocalcinosis in Chinese subjects in Beijing than in white subjects in the United States: the Beijing Osteoarthritis Study. AB - OBJECTIVE: Chondrocalcinosis, which can promote joint inflammation and cartilage degeneration, is highly prevalent in elderly white subjects. Data on its prevalence are scarce in other ethnic populations. This study was undertaken to compare the prevalence of chondrocalcinosis in Chinese subjects with that in white subjects. METHODS: We recruited a random sample of Beijing residents ages >or=60 years. Participants underwent standard weight-bearing anteroposterior knee radiography and posteroanterior hand radiography using the protocols developed in the Framingham Osteoarthritis Study. Radiographic chondrocalcinosis was defined as present in a knee or wrist when there was evidence of definite linear cartilage calcification. We compared the prevalence of chondrocalcinosis in Chinese subjects with that in white subjects using age-standardized prevalence ratios. We used identical methods to collect samples of tap water from 2 cities and measured their levels of calcium, magnesium, and phosphate in the same laboratory. RESULTS: Chinese subjects had a much lower prevalence of knee chondrocalcinosis (1.8% in men, 2.7% in women) than did white subjects (6.2% in men, 7.7% in women), with the age-standardized prevalence ratio being 0.34 (95% confidence interval [95% CI] 0.20-0.54) and 0.43 (95% CI 0.31-0.59) in men and women, respectively. Wrist chondrocalcinosis was rare in elderly Chinese subjects (prevalence 0.3% in men and 1.0% in women), with the age-standardized prevalence ratio being 0.06 (95% CI 0.01-0.18) in Chinese men and 0.18 (95% CI 0.10-0.30) in Chinese women. Calcium levels in the tap water in Beijing were 15-fold higher than in Framingham, whereas no difference was found in magnesium and phosphate levels. CONCLUSION: Knee chondrocalcinosis and wrist chondrocalcinosis are far less common in Chinese subjects in Beijing than in US white subjects in Framingham, Massachusetts. Given the current lack of understanding of the etiology of chondrocalcinosis, further epidemiologic studies of the impact of genetic and environmental factors on occurrence of chondrocalcinosis are indicated. PMID- 17075854 TI - Presentation of 3D isotropic imaging data for optimal viewing. AB - Three-dimensional (3D) isotropically sampled data facilitates retrospective viewing of arbitrarily aligned image planes and simplifies automated computer analysis of 3D structures. However, compared to acquisitions that employ thick slices, MRI acquisitions with isotropic sampling have a lower signal-to-noise ratio (SNR). A rater study was performed to examine viewer preference for the trade-off between SNR and the partial-volume effect of through-plane averaging. A robust preference for a 5:1 ratio in through-plane to in-plane was found for a typical SNR of 11. PMID- 17075855 TI - Functional characterization of an orphan nuclear receptor, Rev-ErbAalpha, in chondrocytes and its potential role in osteoarthritis. AB - OBJECTIVE: To evaluate the expression and function of the orphan nuclear receptor Rev-ErbAalpha in articular cartilage and to investigate its role in osteoarthritis (OA). METHODS: Expression of Rev-ErbAalpha was analyzed at both the messenger RNA and protein levels in human and bovine articular cartilage and chondrocytes by real-time polymerase chain reaction (TaqMan) and immunocytochemical techniques. The effects of cartilage catabolic and anabolic agents on the expression of Rev-ErbAalpha were evaluated by TaqMan analysis. Overexpression was achieved by either adenoviral transduction or treatment with a peroxisome proliferator-activated receptor alpha agonist, whereas expression was suppressed by antisense oligonucleotides. RESULTS: Among the 48 known nuclear receptors, Rev-ErbAalpha was found to be the most highly expressed in OA cartilage. It is known to function as a transcription repressor. Treatment of articular chondrocytes with known catabolic agents resulted in the induction of Rev-ErbAalpha, whereas stimulation with anabolic agents led to a decrease in expression. Overexpression of the nuclear receptor was associated with an increase in the expression of matrix-degrading enzymes such as matrix metalloproteinase 13 and aggrecanase. In contrast, a decrease in Rev-ErbAalpha expression led to a concomitant reduction in the activity of matrix-degrading enzymes. CONCLUSION: This study is the first to demonstrate that Rev-ErbAalpha is highly expressed in OA articular chondrocytes and that its expression is modulated by known cartilage catabolic and anabolic stimuli. We also demonstrated that modulation of Rev-ErbAalpha expression in chondrocytes may be a novel means of regulating the expression and production of multiple matrix-degrading enzymes. These observations suggest that Rev-ErbAalpha may be a novel therapeutic target for OA. PMID- 17075856 TI - The effect of O2 tension on pH homeostasis in equine articular chondrocytes. AB - OBJECTIVE: To determine the effects of varying O(2) on pH homeostasis, based on the hypothesis that the function of articular chondrocytes is best understood at realistic O(2) tensions. METHODS: Cartilage from equine metacarpophalangeal/tarsophalangeal joints was digested with collagenase to isolate chondrocytes, and then loaded with the pH-sensitive fluorophore 2',7'-bis 2-(carboxyethyl)-5(6)-carboxylfluorescein. The radioisotope(22)Na(+) was used to determine the kinetics of Na(+)/H(+) exchange (NHE) and the activity of the Na(+)/K(+) pump, and ATP levels were assessed with luciferin assays. Levels of reactive oxygen species (ROS) were determined using 2',7'-dichlorofluorescein diacetate. RESULTS: The pH homeostasis was unaffected when comparing tissue maintained at 20% O(2) (the level in water-saturated air at 37 degrees C) with that at 5% O(2) (which approximates the normal level in healthy cartilage); however, an O(2) tension of <5% caused a fall in intracellular pH (pH(i)) and slowed pH(i) recovery following acidification, an effect mediated via inhibition of NHE activity (likely through acid extrusion by NHE isoform 1). The Na(+)/K(+) pump activity and intracellular ATP concentration were unaffected by hypoxia, but the levels of ROS were reduced. Hypoxic inhibition of NHE activity and the reduction in ROS levels were reversed by treatment with H(2)O(2), Co(2+), or antimycin A. Treatment with calyculin A also prevented hypoxic inhibition of NHE activity. CONCLUSION: The ability of articular chondrocytes to carry out pH homeostasis is compromised when O(2) tensions fall below those normally experienced, via inhibition of NHE. The putative signal is a reduction in levels of ROS derived from mitochondria, acting via altered protein phosphorylation. This effect is relevant to both physiologic and pathologic states of lowered O(2), such as in chronic inflammation. PMID- 17075857 TI - Theoretical and experimental investigation of the VASO contrast mechanism. AB - Vascular space occupancy (VASO)-dependent functional MRI (fMRI) is a blood nulling technique capable of generating microvascular cerebral blood volume (CBV) weighted images. It is shown that at high magnetic field (3.0T) and high spatial resolution (1.89 x 1.89 x 3 mm(3)), the VASO signal changes are too large (6-7%) to originate from CBV effects alone. Additional contributions are investigated theoretically and experimentally as a function of MRI parameters (TR and TE), as well as the signal-to-noise ratio, (SNR) and spatial resolution. First, it is found that an arterial spin labeling (ASL) contribution causes large negative VASO signal changes at short TR. Second, even at high fMRI spatial resolution, CSF volume contributions (7-13%) cause VASO signal changes to become more negative, most noticeably at long TR and TE. Third, white matter (WM) effects reduce signal changes at lower spatial resolution. The VASO technique has been tested using different stimulus paradigms and field strengths (1-3), giving results consistent with comparable tasks investigated using BOLD and cerebral blood flow (CBF)-based techniques. Finally, simulations show that a mixture of fresh and steady-state blood may significantly alter signal changes at short TR (< or =3 s), permitting larger VASO signal changes than expected under pure steady-state conditions. Thus, many competing effects contribute to VASO contrast and care should be taken during interpretation. PMID- 17075858 TI - Large-scale gene expression profiling reveals major pathogenetic pathways of cartilage degeneration in osteoarthritis. AB - OBJECTIVE: Despite many research efforts in recent decades, the major pathogenetic mechanisms of osteoarthritis (OA), including gene alterations occurring during OA cartilage degeneration, are poorly understood, and there is no disease-modifying treatment approach. The present study was therefore initiated in order to identify differentially expressed disease-related genes and potential therapeutic targets. METHODS: This investigation consisted of a large gene expression profiling study performed based on 78 normal and disease samples, using a custom-made complementary DNA array covering >4,000 genes. RESULTS: Many differentially expressed genes were identified, including the expected up regulation of anabolic and catabolic matrix genes. In particular, the down regulation of important oxidative defense genes, i.e., the genes for superoxide dismutases 2 and 3 and glutathione peroxidase 3, was prominent. This indicates that continuous oxidative stress to the cells and the matrix is one major underlying pathogenetic mechanism in OA. Also, genes that are involved in the phenotypic stability of cells, a feature that is greatly reduced in OA cartilage, appeared to be suppressed. CONCLUSION: Our findings provide a reference data set on gene alterations in OA cartilage and, importantly, indicate major mechanisms underlying central cell biologic alterations that occur during the OA disease process. These results identify molecular targets that can be further investigated in the search for therapeutic interventions. PMID- 17075859 TI - Amyloid contained in the knee joint meniscus is formed from apolipoprotein A-I. AB - OBJECTIVE: To determine the chemical nature of amyloid deposits found in knee joint menisci. METHODS: Amyloid was extracted from the menisci of 3 adults who underwent knee joint replacement surgery. The primary structural features of the purified proteins were determined by sequential Edman degradation and tandem mass spectrometry (MS/MS). Tissue specimens were also subjected to in situ hybridization analysis, as well as complementary DNA cloning by reverse transcriptase-polymerase chain reaction (RT-PCR). Additionally, specimens from these 3 patients, as well as other patients with amyloid in the knee joint menisci, were examined immunohistochemically. RESULTS: Amino acid sequence and MS/MS analyses of the extracts revealed the presence of 60-77-residue components identical to the N-terminal portion of apolipoprotein A-I (Apo A-I). The Apo A-I nature of the amyloid was confirmed by the demonstration that the green birefringent congophilic deposits in the 7 meniscus samples were recognized by an anti-human Apo A-I antibody. That the meniscus itself was the source of the amyloidogenic protein was evidenced through Southern blot analysis, in which an Apo A-I product was generated by RT-PCR from synovial tissue, and further, by the demonstration that the cytoplasm of chondrocytes reacted with the specific Apo A I probe used for in situ hybridization and was immunostained by the anti-Apo A-I antiserum. CONCLUSION: Amyloid in the knee joint menisci is formed from Apo A-I that is produced by chondrocytes within the meniscal cartilage. This entity represents yet another localized form of amyloidosis associated with the aging process and may be of pathophysiologic import. PMID- 17075860 TI - Modified cellulose II powder: preparation, characterization, and tableting properties. AB - The reaction of UICEL-A/102, a cellulose II powder recently prepared from Avicel(R) PH-102 by treatment with an aqueous sodium hydroxide solution, with glutaraldehyde in 0.01 N HCl has been investigated to improve its binder properties, without adversely affecting the rapid disintegration characteristic. The results showed that UICEL-A/102 and glutaraldehyde when reacted in a 1:0.6 weight ratio at 100 degrees C for 8.5 h produces a product, (hereinafter referred to as UICEL-XL), that, compared to UICEL-A/102, had a lower degree of polymerization, higher crystallinity, lower bulk density, lower tapped density, and higher porosity. Further, it showed lower yield pressure and higher crushing strength, and tensile strength values, indicating that UICEL-XL is more compressible and compactable than the starting material, UICEL-A/102. A comparison of "in-die" and "out-of-die" Heckel data indicated UICEL-XL to be less elastic than UICEL-A/102. Both UICEL-XL and UICEL-A/102 showed similar moisture sorption isotherms, and their compacts disintegrated rapidly in water. In conclusion, the glutaraldehyde-treated cellulose II powder not only serves as good a disintegrant as the untreated cellulose powder but also possesses superior binder properties. PMID- 17075861 TI - Impaired skeletal development in interleukin-6-transgenic mice: a model for the impact of chronic inflammation on the growing skeletal system. AB - OBJECTIVE: To identify the mediator responsible for the impact of chronic inflammation on skeletal development in children (bone loss, defective peak bone mass accrual, stunted growth), we evaluated the effects of chronic interleukin-6 (IL-6) overexpression on the skeletons of growing prepubertal mice. METHODS: We studied IL-6-transgenic mice that had high circulating IL-6 levels since birth. Trabecular and cortical bone structure were analyzed by microcomputed tomography. Epiphyseal ossification, growth plates, and calvariae were studied by histology/histomorphometry. Osteoclastogenesis, osteoblast function/differentiation, and the effects of IL-6 on bone cells were studied in vitro. Osteoblast gene expression was evaluated by reverse transcriptase polymerase chain reaction. The mineral apposition rate was evaluated dynamically in cortical bone by in vivo double fluorescence labeling. RESULTS: In prepubertal IL-6-transgenic mice, we observed osteopenia, with severe alterations in cortical and trabecular bone microarchitecture, as well as uncoupling of bone formation from resorption, with decreased osteoblast and increased osteoclast number and activity. Increased osteoclastogenesis and reduced osteoblast activity, secondary to decreased precursor proliferation and osteoblast function, were present. IL-6 transgenic mice also showed impaired development of growth plates and epiphyseal ossification centers. Intramembranous and endochondral ossification and the mineral apposition rate were markedly affected, showing the presence of defective ossification. CONCLUSION: Chronic overexpression of IL-6 alone induces a skeletal phenotype closely resembling growth and skeletal abnormalities observed in children with chronic inflammatory diseases, pointing to IL-6 as a pivotal mediator of the impact of chronic inflammation on postnatal skeletal development. We hypothesize that IL-6-modifying drugs may reduce skeletal defects and prevent the growth retardation associated with these diseases. PMID- 17075862 TI - Patients with juvenile psoriatic arthritis comprise two distinct populations. AB - OBJECTIVE: Psoriatic arthritis (PsA) in children is clinically heterogeneous. We examined a large population of children with juvenile PsA for evidence of phenotypic clustering that could suggest the presence of distinct clinical entities. METHODS: We reviewed the medical records of 139 patients meeting the Vancouver criteria for juvenile PsA. To identify segregation into phenotypic groups, we compared younger patients with their older counterparts and subjected the whole population to 2-step cluster analysis. RESULTS: Among patients with juvenile PsA, the age at onset is biphasic, with peaks occurring at approximately 2 years of age and again in late childhood. Compared with children ages 5 years and older, younger patients are more likely to be female, exhibit dactylitis and small joint involvement, and express antinuclear antibodies. Progression to polyarticular disease (>or=5 joints) is more common in younger children, although joint involvement remains oligoarticular in the majority of children. In contrast, older patents tend to manifest enthesitis, axial joint disease, and persistent oligoarthritis. Uveitis is equally represented in both age groups. Despite a higher utilization of methotrexate therapy, younger patients required, on average, more than twice as long to achieve clinical remission (23 months versus 9.2 months; P = 0.044). Cluster analysis identified largely overlapping subgroups but suggested that the presence of dactylitis, rather than age, has the greatest capacity to predict essential features of the clinical phenotype. CONCLUSION: Juvenile PsA comprises 2 distinct populations of patients. Although the pathophysiologic correlate of this finding remains undefined, future studies should avoid the assumption that PsA in childhood constitutes a single etiologic entity. PMID- 17075863 TI - Long-term outcome and prognostic factors in enthesitis-related arthritis: a case control study. AB - OBJECTIVE: To compare the clinical, functional, and radiographic outcomes in patients with enthesitis-related arthritis (ERA) with those in patients with other subtypes of juvenile idiopathic arthritis (JIA) and healthy controls, and to determine genetic markers, patient characteristics, and early disease variables that predict the development of remission, sacroiliitis, and physical limitations in ERA. METHODS: Fifty-five children with ERA who were first admitted to Rikshospitalet Medical Center between 1980 and 1985 were studied. Patients with oligoarthritis or polyarthritis who were admitted during the same period (n = 55) and individuals from a national population registry (n = 55) were matched for sex and age and used as controls. Health status was assessed after a median of 15.3 years of disease (range 11.7-21.9 years) and, in some patients, was reassessed after a median of 23.0 years (range 19.7-29.4 years) of disease, by use of the 36-item Short Form health survey and the Health Assessment Questionnaire. Clinical and radiographic examinations were performed at the 15 year followup visit. Variables relating to the onset of disease were retrospectively obtained by chart review. HLA alleles were determined by genotyping and serologic testing. RESULTS: Patients with ERA had lower levels of physical functioning, poorer physical health, and more bodily pain compared with patients with oligoarthritis or polyarthritis (after a median of 15.3 and a median of 23.0 years) and normal controls (after a median of 15.3 years). Among patients with ERA, remission occurred in 44%, sacroiliitis was observed in 35%, and reduced spinal flexion was observed in 75%. Predictors of failure to attain disease remission included the following: ankylosing spondylitis (AS) in a first degree relative, the presence of HLA-DRB1*08, and ankle arthritis within the first 6 months. HLA-DPB1*02 was a protective factor, whereas a persistently elevated erythrocyte sedimentation rate (ESR), and hip arthritis within the first 6 months were risk factors for sacroiliitis. Female sex, a family history of AS, and high numbers of affected joints within the first 6 months predicted poor physical health status after 23 years. Male sex was associated with reduced anterior flexion of the spine. CONCLUSION: In this study, patients with ERA had poorer physical outcomes compared with patients with oligoarticular or polyarticular JIA and controls from the general population. A family history of related diseases, sex, the presence of HLA-DRB1*08, the absence of HLA-DPB1*02, a persistently elevated ESR, early hip or ankle arthritis, and high numbers of affected joints were predictors of an unfavorable outcome. PMID- 17075864 TI - Identification of immunodominant CD4+ T cell epitopes in patients with Yersinia induced reactive arthritis by cytometric cytokine secretion assay. AB - OBJECTIVE: In reactive arthritis (ReA), a bacteria-specific T cell response to the triggering microbe is detected in synovial fluid. So far, direct characterization of bacteria-specific T cells and identification of the immunodominant fine specificities remain difficult due to the lack of appropriate techniques. The aim of the present study was to directly determine the fine specificity of CD4+ T cells specific to ReA-associated bacteria-derived protein. METHODS: In 2 patients with Yersinia-induced ReA, live Yersinia Hsp60-specific CD4+ T cells were directly isolated from synovial fluid after stimulation with Yersinia-derived protein Hsp60 using a cytometric cytokine secretion assay. Generated short-term T cell lines were then tested in vitro for their peptide epitope specificity. Also, direct cross-reactivity of one line with Chlamydia- and human-derived Hsp60 was assessed. RESULTS: Generated short-term CD4+ T cell lines were highly antigen-specific and revealed single immunodominant peptide epitopes that were confirmed by direct testing with single peptides in both peripheral blood and synovial fluid cells. Yersinia Hsp60-specific T cells of one patient cross-reacted directly with human Hsp60. CONCLUSION: Our results demonstrate the feasibility of direct assessment of live, potentially pathogenic, antigen-specific interferon-gamma+ CD4+ T cells taken from inflammatory lesions of patients with rheumatic diseases such as ReA. This might have implications not only regarding pathogenesis, but also in the design of new immunotherapies. PMID- 17075865 TI - Biodistribution and pharmacokinetic analysis of long-circulating thiolated gelatin nanoparticles following systemic administration in breast cancer-bearing mice. AB - The objective of the present study was to modify thiolated gelatin nanoparticles with poly(ethylene glycol) (PEG) chains and examine their long circulating and tumor-targeting properties in vivo in an orthotopic a human breast adenocarcinoma xenograft model. The crosslinked nanoparticle systems were characterized to have a size of 150-250 nm with rapid payload release properties in a highly reducing environment. Upon PEG modification, the nanoparticle size increased to 300-350 nm in diameter. The presence of PEG chains on the surface was confirmed by characterization with electron spectroscopy for chemical analysis. The in vivo long-circulating potential, biodistribution and passive tumor targeting of the controls, and PEG-modified thiolated gelatin nanoparticles were evaluated by injecting indium-111 (111In)-labeled nanoparticles into breast tumor (MDA-MB-435) bearing nude mice. Upon modification with PEG, the nanoparticles were found to have longer circulation times, with the plasma and tumor half-lives of 15.3 and 37.8 h, respectively. The results also showed preferential localization of thiolated nanoparticles in the tumor mass. The resulting nanoparticulate systems with long circulation properties could be used to target encapsulated drugs and genes to tumors passively by utilizing the enhanced permeability and retention effect of the tumor vasculature. PMID- 17075866 TI - Expression of claudins in murine tooth development. AB - Claudins belong to a family of transmembrane proteins that were identified as components of tight junction strands. We carried out comparative in situ hybridization analysis of 11 claudin genes (claudin1 - claudin11) during murine odontogenesis from the formation of the epithelial thickening to the cytodifferentiation stage. We identify dynamic spatiotemporal expression of 9 of the 11 claudins. At the early bell stage, two claudins (claudin1 and 4) are specifically expressed in stratum intemedium, whereas only one claudin is expressed in each of the preameloblasts (claudin2) and preodontoblasts (claudin10). At the bud stage, when the first epithelial differentiation pathways are being established, localized expression of six claudins (claudin1, 3, 4, 6, 7, and 10) identify spatial specific interactions, suggesting a hitherto unobserved complexity of epithelial organization, within the early tooth primordium. PMID- 17075867 TI - Development and validation of a preclinical food effect model. AB - A preclinical canine model capable of predicting a compound's potential for a human food effect was developed. The beagle dog was chosen as the in vivo model. A validation set of compounds with known propensities for human food effect was studied. Several diets were considered including high-fat dog food and various quantities of the human FDA meal. The effect of pentagastrin pretreatment was also investigated. The high-fat dog food did not predict human food effect and was discontinued from further evaluation. The amount of FDA meal in the dog was important in the overall prediction of the magnitude of human food effect. Fed/fasted Cmax and AUC ratios using a 50-g aliquot of the FDA meal in the dog were in the closest qualitative agreement to human data. Pentagastrin pretreatment did not affect the AUC in the fed state, but increased the fasted AUC for weakly basic compounds. Pentagastrin pretreatment and a 50-g aliquot of the FDA meal in the dog predicted the human food effect for a validation set of compounds. This model, which is intended for compound screening, will be helpful for determining food effect as a liability when compounds progress from discovery to clinical development. PMID- 17075868 TI - Modifying drug release and tablet properties of starch acetate tablets by dry powder agglomeration. AB - In this study three model drugs (N-acetyl-D-glucosamine (NAG), anhydrous caffeine, and propranolol hydrochloride) were agglomerated with starch acetate (SA) by mixing the binary powders on a stainless steel (SS) plate. Agglomeration was induced by triboelectrification of the particles during mixing, and it was evaluated as a method to achieve controlled drug release rate. These agglomerates, mixed with different amounts of a disintegrant, were compressed into tablets whose dissolution characteristics were determined. Triboelectric measurements showed that when the drugs were in contact with SS, charges of the opposite polarity were generated to SA (+) and caffeine and NAG (-) promoting adhesion. Instead, propranolol HCl was charged with the same polarity as SA. SEM micrographs showed that smaller caffeine particles, in spite of their larger negative charge, agglomerated less efficiently with SA than larger NAG particles. This emphasizes the importance of particle size in the agglomeration process. Propranolol HCl did not form agglomerates with SA since their particle sizes and charges were identical. As a result, agglomeration of powders prior to tablet compression allows for modification and control of the release rate of the drugs from the SA matrix tablets as well as the tensile strength of the tablets. PMID- 17075869 TI - Hot-melt extrusion for enhanced delivery of drug particles. AB - With the recent advent of nanotechnology for pharmaceutical applications, drug particle engineering is the focus of increasing interest as a viable approach for overcoming solubility limitations of poorly water-soluble drugs. Although these particle engineering techniques have been proven successful for enhancing the dissolution properties of many poorly water-soluble drugs, there are limitations associated with them such as particle aggregation, morphological instability, and poor wettability. The aim of this study was to demonstrate a processing technique in which hot-melt extrusion (HME) is utilized to overcome these limitations. Micronized particles of amorphous itraconazole (ITZ) stabilized with PVP or HPMC were produced and subsequently melt extruded with poloxamer 407 and PEO 200 M to deaggregate and disperse the particles into the hydrophilic polymer matrix. Differential scanning calorimetry, X-ray diffraction, and scanning electron microscopy were used to demonstrate that the HME process did not alter the properties of the micronized particles. Dissolution testing conducted at sink conditions revealed that the dissolution rate of the micronized particles was improved by HME due to particle deaggregation and enhanced wetting. Supersaturation dissolution testing demonstrated that the ITZ-HPMC micronized particle extrudates provided superior supersaturation of ITZ compared to the ITZ PVP micronized particle extrudates. Supersaturation dissolution testing incorporating a pH change (from pH 1.2 to 6.8 at 2 h) revealed that neither micronized particle extrudate formulation significantly reduced the rate of ITZ precipitation from supersaturated solution once pH was increased. Moreover, the two extrudate formulations performed very similarly when only considering dissolution testing from just before pH adjustment through the duration of testing at neutral pH. From oral dosing of rats, it was determined that the two extrudate formulations performed similarly in vivo as confirmed by their statistically equivalent AUC values. By correlating the results of supersaturation dissolution testing with pH change to the in vivo AUC, it appears that rapid precipitation of ITZ occurs upon entrance into the more neutral pH environment of the small intestine resulting in a brief opportunity for absorption. This suggests that perhaps the optimum formulation approach for ITZ is to control drug release so as to retard precipitation as pH is increased and extend the absorption window in the small intestine. PMID- 17075870 TI - Validating a prognostic model. PMID- 17075871 TI - Low predictive accuracy of the Kattan postoperative nomogram for renal cell carcinoma recurrence in a population of French patients. AB - BACKGROUND: The aim of the current study was to establish the predictive accuracy of the Kattan postoperative nomogram for nonmetastatic renal cell carcinoma (RCC) by comparing predictions with actual disease recurrence in patients who underwent surgery in a single center in France. METHODS: Between 1985 and 2000, 844 patients were treated for RCC. The following data were collated: age, symptoms, histology, tumor size, grade, TNM 1997 stage, recurrence, and progression. For each patient a prognostic score (predicted probability) for recurrence-free survival (RFS) at 5 years was calculated using the Kattan nomogram. The discriminating ability of the model was assessed by Harrell's concordance index (c-index). Bootstrapping was used to assess confidence intervals. Furthermore, survival was then estimated by the Kaplan-Meier method and Cox proportional hazards regression analysis. RESULTS: In all, 565 patients (median age, 62 years) were included. At the time of the last follow-up, 81 patients had died and 101 had experienced RCC recurrence. The c-index for RFS (Kattan nomogram) was only 0.607 (95% confidence interval [CI]: 0.576-0.635). The 5-year RFS rate and cancer specific survival rate were 81.5% and 84.7%, respectively. Of the 4 variables included in the nomogram, only TNM stage was associated with recurrence in a multivariate analysis (Cox analysis) (P = .022). CONCLUSIONS: There was a discrepancy between predicted RFS as estimated by the Kattan nomogram and the likelihood of being recurrence-free at 5 years according to the Cox analysis in the current population of patients. However, until new dynamic models become available clinicians may still improve their predictive ability by using the current nomogram. PMID- 17075872 TI - Start the clock! Circadian rhythms and development. AB - The contribution of timing cues from the environment to the coordination of early developmental processes is poorly understood. The day-night cycle represents one of the most important, regular environmental changes that animals are exposed to. A key adaptation that allows animals to anticipate daily environmental changes is the circadian clock. In this review, we aim to address when a light-regulated circadian clock first emerges during development and what its functions are at this early stage. In particular, do circadian clocks regulate early developmental processes? We will focus on results obtained with Drosophila and vertebrates, where both circadian clock and developmental control mechanisms have been intensively studied. PMID- 17075874 TI - Assessment of symptom experience in patients undergoing hepatic resection or ablation. AB - BACKGROUND: Quality of life (QOL) currently is considered both clinically meaningful and biologically important for patient outcome and is considered as important as disease-free and overall survival. Thus, the objective of the current study was to evaluate the QOL symptoms of patients who underwent major hepatic resection, minor hepatic resection, and ablation for primary or metastatic cancer to the liver. METHODS: From October 2002 to June 2004, 40 patients who underwent either hepatic ablation or resection were enrolled. Patients were assessed at 5 time points (the initial visit, the initial postoperative visit, and visits at 6 weeks, 3 months, and 6 months) by questionnaires of the Functional Assessment in Cancer Therapy (FACT) core instrument with the Hepatobiliary subscale (FACT-Hep), the FACT Hepatobiliary Symptom Index (FHSI-8), the Profile of Mood States (POMS), the European Organization for Research and Treatment of Cancer (EORTC) QOL questionnaire (QLQ) for patients with pancreatic cancer (QLQ-PAN), and the general core EORTC QLQ. RESULTS: The patients enrolled included 20 men and 20 women with a median age of 62 years (range, 41-77 years), including 24 patients who underwent major hepatectomy, 8 patients who underwent minor hepatectomy, and 8 patients who underwent ablation. An evaluation of the FACT Physical, Social, Emotional, and Functional subscales demonstrated no differences at the initial or first postoperative visits. However, at 6 weeks, both the Physical (P = .0455) and Functional (P = .0372) scores were significantly worse for the major hepatectomy group. At 3 months, all QOL parameters were similar. Similar differences were observed at 6 weeks for the FHSI-8 (P = .02), POMS (P = .007), QLQ-PAN (P = .04), and EORTC (P = .003) with the resolution of this difference at 3 months. CONCLUSIONS: There was little difference in QOL between patients who underwent major hepatic resection, minor hepatic resection, and hepatic ablation. Patients who underwent major hepatectomy demonstrated a worse QOL at 6 weeks compared with patients who underwent minor hepatic resection and hepatic ablation, with the resolution of this difference and significant improvements observed in all 3 groups at 3 months. PMID- 17075873 TI - Characterization of Xenopus digits and regenerated limbs of the froglet. AB - Xenopus has 4 and 5 digits in a forelimb and hindlimb, respectively. It is thought that their limbs and digits develop in Xenopus by mechanisms that are almost conserved from amphibians to higher vertebrates. This is supported by some molecular evidence. The 5'hoxd genes are convenient marker genes for characterizing digits in the chick and mouse. The anteriormost digit is characterized by being hoxd13-positive and hoxd12 (hoxd11)-negative in the chick and mouse. In this study, we revealed that the anteriormost digit of the Xenopus forelimb is hoxd13-positive and hoxd11-positive, that is, a more posterior character than digit I. The order of formation of digit cartilages also suggested that Xenopus forelimb digit identity is II to V, not I to IV. We have also been interested in the relationship between digit identity and shh. The anteriormost digit develops in a shh-independent way. A limb treated with cyclopamine (a shh inhibitor) has a gene expression pattern (hoxd11-negative) similar to that in shh deficient mice, suggesting that a hindlimb treated with cyclopamine has a digit I character. However, a Xenopus froglet regenerate (spike), which lacks shh expression during its regeneration process, does not have such an expression pattern, being hoxd11-positive. We investigated hoxd11 transcriptions in blastemas that formed in the anteriormost and posteriormost digits, and we found that the blastemas have different hoxd11 expression levels. These findings suggest that the froglet limb blastema does not have a mere digit I character in spite of shh defectiveness and that the froglet limb blastema recognizes its positional differences along the anterior-posterior axis. PMID- 17075875 TI - Differential expression of anti-Mullerian hormone (amh) and anti-Mullerian hormone receptor type II (amhrII) in the teleost medaka. AB - In mammals, the anti-Mullerian hormone (Amh) is responsible for the regression of the Mullerian ducts; therefore, Amh is an important factor of male sex differentiation. The amh gene has been cloned in various vertebrates, as well as in several teleost species. To date, all described species show a sexually dimorphic expression of amh during sex differentiation or at least in differentiated juvenile gonads. We have identified the medaka amh ortholog and examined its expression pattern. Medaka amh shows no sexually dimorphic expression pattern. It is expressed in both developing XY male and XX female gonads. In adult testes, amh is expressed in the Sertoli cells and in adult ovaries in granulosa cells surrounding the oocytes, like in mammals. To better understand the function of amh, we cloned the anti-Mullerian hormone receptor type II (amhrII) ortholog and compared its expression pattern with amh, aromatase (cyp19a1), and scp3. During gonad development, amhrII is coexpressed with medaka amh in somatic cells of the gonads and shows no sexually dimorphic expression. Only the expression level of the Amh type II receptor gene was decreased noticeably in adult female gonads. These results suggest that medaka Amh and AmhrII are involved in gonad formation and maintenance in both sexes. PMID- 17075876 TI - Nuclear receptor NR5A2 is required for proper primitive streak morphogenesis. AB - NR5A2, also known as liver receptor homologue 1 (LRH-1) and fetoprotein transcription factor (FTF), is an orphan nuclear receptor involved in the regulation of cholesterol metabolism and steroidogenesis in the adult. NR5A2 was also shown to be expressed during early mouse embryogenesis. Consistent with its early expression pattern, a targeted disruption of this gene leads to embryonic lethality around the gastrulation period. To characterize the embryonic phenotype resulting from NR5A2 loss of function, we undertook morphological and marker gene analyses and showed that NR5A2-/- embryos display growth retardation, epiblast disorganization, a mild embryonic-extraembryonic constriction, as well as abnormal thickening of the proximo-posterior epiblast. We demonstrated that, although initial specification of the anterior-posterior axis occurred in the absence of NR5A2, primitive streak formation was impaired and neither embryonic nor extraembryonic mesoderm was generated. Moreover, although the visceral endoderm does not show major morphological abnormalities in NR5A2-/- embryos, a decrease in the expression level of HNF4 and GATA4 was observed. Aggregation experiments demonstrated that, in the presence of wild-type tetraploid cells, NR5A2 mutant cells in the epiblast are capable of undergoing normal gastrulation. Therefore, our results suggest a requirement for NR5A2 in extraembryonic tissues and identify a novel role of this gene in proper primitive streak morphogenesis. PMID- 17075877 TI - Outcome of gastric cancer patients after successful gastrectomy: influence of the type of recurrence and histology on survival. AB - BACKGROUND: The effect of the location of disease recurrence after curative (R0) gastrectomy on patient survival has not been elucidated. The authors hypothesized that the location of recurrence would have a significant influence on survival. METHODS: Medical records of all patients who received treatment for gastric cancer at The University of Texas M. D. Anderson Cancer Center between 1985 and 1998 were reviewed. Patients who underwent R0 resection for gastric cancer and subsequently developed localized (anastomotic) recurrence (LR), lymph node (regional) recurrence (NR), or distant metastases (DM) were analyzed for overall survival (OS). All study factors were entered into a Cox proportional hazards model to provide multivariate hazard ratios. The model was adjusted for the effects of primary site of recurrence, histologic grade, patient age, and location of the primary tumor. RESULTS: This retrospective analysis included 227 consecutive patients. The median survival of patients who developed NR (11 months) was similar to that of patients who developed LR (10 months), but both groups had significantly longer median survival compared with patients who developed DM (7 months; log-rank P = .03). Patients who had well differentiated or moderately differentiated tumors had a longer OS (11 months) than patients who had poorly differentiated tumors (8 months; log-rank P = .02). In this cohort, location of the primary cancer and age at recurrence had no significant impact on OS. CONCLUSIONS: The data from this study suggested that, among patients who undergo R0 gastrectomy for gastric cancer, LR and NR versus DM should be considered a valid stratification factor for randomized trials based on significant differences in survival. Determining whether this stratification should apply to histologic differentiation will require further investigation in a larger multicenter cohort. PMID- 17075878 TI - A Phase I study of bortezomib plus irinotecan in patients with advanced solid tumors. AB - BACKGROUND: The authors conducted a Phase I dose-finding trial to study the use of combined bortezomib plus irinotecan in patients with advanced solid tumors. METHODS: Patients who had received >/=1 prior chemotherapy regimen were eligible. Patients received bortezomib (1.0 mg/m(2), 1.3 mg/m(2), or 1.5 mg/m(2)) on Days 1, 4, 8, and 11 and received irinotecan (from 50 mg/m(2) to 125 mg/m(2)) on Days 1 and 8 of each 21-day cycle for a maximum of 8 cycles. Bortezomib followed irinotecan on coadministration days in Cycle 1 and Cycles 3 through 8 but preceded irinotecan in Cycle 2 to assess the effect of administration sequence on bortezomib pharmacodynamics. RESULTS: Fifty-one enrolled patients with malignancies, including colorectal cancer (n = 23 patients), lung cancer (n = 6 patients), gastroesophageal cancer (n = 6 patients), and pancreatic cancer (n = 3 patients), received >/=1 dose of study drug. Nausea, vomiting, and diarrhea were the principal dose-limiting toxicities and led to the maximum tolerated doses of 1.3 mg/m(2) bortezomib and 125 mg/m(2) irinotecan. The most common grade >/=3 bortezomib-related nonhematologic adverse events were fatigue (n = 5 episodes), diarrhea (n = 4 episodes), and nausea (n = 4 episodes). grade >/=3 bortezomib related hematologic adverse events included neutropenia (n = 6 episodes) and thrombocytopenia (n = 4 episodes) and rarely were dose limiting. Of 34 evaluable patients, no objective responses according to the Response Evaluation Criteria in Solid Tumors were seen; 10 patients achieved stable disease. The degree of proteasome inhibition in whole blood indicated that the biologic activity of bortezomib was unaffected by irinotecan coadministration. CONCLUSIONS: The results of this Phase I study in patients with solid tumors indicated that bortezomib at a dose of 1.3 mg/m(2) on Days 1, 4, 8, and 11 plus irinotecan at a dose of 125 mg/m(2) on Days 1 and 8 every 21 days were the recommended Phase II doses. PMID- 17075879 TI - Phase II study of lenalidomide in patients with metastatic renal cell carcinoma. AB - BACKGROUND: Lenalidomide (LEN) is a structural and functional analogue of thalidomide that has demonstrated enhanced immunomodulatory properties and a more favorable toxicity profile. A Phase II, open-label study of LEN in patients with metastatic renal cell carcinoma (RCC) was conducted to determine its safety and clinical activity. METHODS: Patients with metastatic RCC received LEN orally at a dose of 25 mg daily for the first 21 days of a 28-day cycle. The primary endpoint was the objective response rate. Time to treatment failure, safety, and survival were secondary endpoints. RESULTS: In total, 28 patients participated in the trial and were included in the current analysis. Three of 28 patients (11%) demonstrated partial responses and continued to be progression-free for >15 months. Eleven patients (39%) had stable disease that lasted >3 months, including 8 patients who had tumor shrinkage. In total, 6 patients (21%) remained on the trial, and 5 additional patients continued to be followed for survival. The median follow-up for those 11 patients was 13.5 months (range, 8.3-17.0 months). The median survival had not been reached at the time of the current report. Serious adverse events included fatigue (11%), skin toxicity (11%), and neutropenia (36%). CONCLUSIONS: LEN demonstrated an antitumor effect in metastatic RCC, as evidenced by durable partial responses. LEN toxicities were manageable. Further studies will be required to assess the overall activity of LEN in patients with metastatic RCC. PMID- 17075880 TI - Mice with mutations in Mahogunin ring finger-1 (Mgrn1) exhibit abnormal patterning of the left-right axis. AB - Mahogunin Ring Finger 1 (Mgrn1) encodes a RING-containing protein with ubiquitin ligase activity that has been implicated in pigment-type switching. In addition to having dark fur, mice lacking MGRN1 develop adult-onset spongy degeneration of the central nervous system and have reduced embryonic viability. Observation of complete situs inversus in a small proportion of adult Mgrn1 mutant mice suggested that embryonic lethality resulted from congenital heart defects due to defective establishment and/or maintenance of the left-right (LR) axis. Here we report that Mgrn1 is expressed in a pattern consistent with a role in LR patterning during early development and that many Mgrn1 mutant embryos show abnormal expression of asymmetrically expressed genes involved in LR patterning. A range of complex heart defects was observed in 20-25% of mid-to-late gestation Mgrn1 mutant embryos and another 20% were dead. This finding was consistent with 46-60% mortality of mutants by weaning age. Our results indicate that Mgrn1 acts early in the LR signaling cascade and is likely to provide new insight into this developmental process as Nodal expression was uncoupled from expression of other Nodal-responsive genes in Mgrn1 mutant embryos. Our work identifies a novel role for MGRN1 in embryonic patterning and suggests that the ubiquitination of MGRN1 target genes is essential for the proper establishment and/or maintenance of the LR axis. PMID- 17075881 TI - Antiviral therapy for hepatitis C virus--associated mixed cryoglobulinemia vasculitis: a long-term followup study. AB - OBJECTIVE: To evaluate the long-term efficacy of anti-hepatitis C virus (HCV) therapy in patients with HCV-associated mixed cryoglobulinemia (HCV-MC) vasculitis and to assess the factors associated with clinical remission of MC. METHODS: This was a single-center study of 72 consecutive patients who received treatment with IFN alfa-2b (3 million IU 3 times a week; n = 32 patients) or PEGylated IFN alfa-2b (PEG-IFN alfa-2b) (1.5 mug/kg/week; n = 40 patients), both in combination with oral ribavirin (600-1,200 mg/day), for at least 6 months. Logistic regression was used to assess factors associated with clinical remission of MC. RESULTS: The mean +/- SD duration of followup after discontinuation of antiviral therapy was 39.7 +/- 24.4 months. Eight deaths (11.1% of patients) occurred during the study, primarily as a result of cardiovascular disease, liver disease, or infection. A complete clinical response of the MC occurred in 45 patients (62.5%), a sustained virologic response occurred in 58.3%, and cryoglobulins cleared in 45.8%. Compared with patients treated with IFN alfa-2b plus ribavirin, those receiving PEG-IFN alfa-2b plus ribavirin had a higher sustained clinical (67.5% versus 56.3%), virologic (62.5% versus 53.1%), and immunologic (57.5% versus 31.3%) response, regardless of HCV genotype and viral load. In multivariate analyses, an early virologic response (odds ratio 3.53 [95% confidence interval 1.18-10.59]) was independently associated with a complete clinical response of MC. A glomerular filtration rate 0.05). Mixed infection with genotype A and D were seen in 3% of the cases. Genotype D was the dominant genotype prevalent in all patient categories. PMID- 17075989 TI - Levels of serologic markers of celiac disease in patients with reflux esophagitis. AB - AIM: To investigate the prevalence of celiac disease serologic markers (antigliadin IgA, IgG, and anti-endomysial IgA) in patients with reflux esophagitis and to detect the relationship between reflux esophagitis and celiac disease (CD). METHODS: This study was performed prospectively between January 2003 and January 2004. Sixty-eight adult reflux esophagitis patients and 40 people as control group for symptoms related with gastrointestinal system were enrolled in this study. The diagnostic work-up included an accurate medical history with gastrointestinal symptoms, routine laboratory measurements, the detection of antibodies against gliadin (IgA and IgG) and endomysium (IgA), and an upper endoscopy with postbulbar biopsy. RESULTS: IgA-AGA and IgG-AGA were positive at 8.8% and 10.3% in patients with reflux esophagitis. In control group, it was found that 10% people had positive IgA-AGA, and 7.5% people had positive IgG-AGA. There was no significant relationship between patients and control group regarding positive IgA-AGA and IgG-AGA. The patients and persons in control group had no positive IgA-EMA. On postbulbar biopsies, no finding was detected concerning celiac disease. There were no symptoms and signs for gluten enteropathy in patients and control group. CONCLUSION: This review supports that an association does not exist between celiac disease and reflux esophagitis. We think these diseases exist independently from each other. PMID- 17075990 TI - Monocytes in systematic inflammatory response syndrome: differences between sepsis and acute pancreatitis. AB - AIM: To unravel the differences between systematic inflammatory response syndrome (SIRS) of acute pancreatitis compared to the same syndrome in sepsis. METHODS: Twenty-five patients were enrolled, 12 with sepsis and 13 acute pancreatitis. After diagnosis 20 mL blood was sampled. Half were assayed for isolation of monocytes and 10 mL was centrifuged for serum test of tumor necrosis factor alpha (TNFalpha and interleukin-6 (IL-6). Half of monocytes were incubated in the presence of patients' serum and supernatants were collected. The other half was treated for estimation of optical photometry under caspase-3 inhibition. TNFalpha and IL-6 were estimated by an enzyme immunoassay. RESULTS: median+/-SE of serum IL-6 in septic patients and acute pancreatitis patients was 192.30+/-35.40 ng/L and 21.00+/-16.05 ng/L, respectively (P<0.01). Respective values of caspase-3 were 0.94+/-0.17 pmol/min 10(4) cells and 0.34+/-0.09 pmol/min 10(4) cells (P<0.05). IL-6 of monocyte supernatants of patients with sepsis was significantly increased after addition of patients' serum, while that of patients with acute pancreatitis did not show significant difference. CONCLUSION: The data have shown that monocyte activity is different between acute pancreatitis and sepsis. This phenomenon might be explained as a different pathway to the pro-inflammatory cytokines release or could be a novel anti-inflammatory response in acute pancreatitis. PMID- 17075992 TI - Hyperbilirubinemia after extracorporeal circulation surgery: a recent and prospective study. AB - AIM: To clarify the incidence and nature of postoperative hyperbilirubinemia in patients after modern extracorporeal circulation, to analyze possible perioperative risk factors, and to elucidate the clinical significance of postoperative hyperbilirubinemia associated mortality and morbidity. METHODS: Between March 2005 and May 2006, three hundred and eighty six consecutive patients undergoing extracorporeal circulation surgery due to a variety of cardiac lesions were investigated prospectively. The incidence of postoperative hyperbilirubinemia was defined as a serum total bilirubin concentration of more than 51 micromol/L. Several perioperative parameters were compared by logistic regression between hyperbilirubinemia and non-hyperbilirubinemia patients to determine possible risk factors contributing to postoperative hyperbilirubinemia and mortality. RESULTS: Overall incidence of postoperative hyperbilirubinemia was 25.3% (98/386). In patients with postoperative hyperbilirubinemia, 56.2% reached peak total bilirubin concentration on the first postoperative day, 33.5% on the second day, and 10.3% on the seventh day. Eighty percent of the increase of total bilirubin resulted from an increase of both conjugated and unconjugated bilirubin. Development of postoperative hyperbilirubinemia was associated with a higher mortality (P<0.01), longer duration of mechanical ventilation (P<0.05) and longer ICU stay time (P<0.05). Preoperative total bilirubin concentration, preoperative right atrium pressure, numbers of valves replaced and of blood transfusion requirement were identified as important predictors for postoperative hyperbilirubinemia. CONCLUSION: Early postoperative hyperbilirubinemia after modern extracorporeal circulation is mainly caused by an increase in both conjugated and unconjugated bilirubin, and is associated with a high mortality. Important contributing factors are the preoperative total bilirubin concentration, preoperative severity of right atrial pressure, numbers of valve replacement procedures, and the amount of blood transfusion requirement during and shortly after surgery. We suggest that postoperative hyperbilirubinemia is a multifactorial process, which is caused by both the impaired liver function of bilirubin transport and the increased production of bilirubin from haemolysis. PMID- 17075991 TI - Efficacy of thymosin alpha-1 and interferon alpha in treatment of chronic viral hepatitis B: a randomized controlled study. AB - AIM: To observe the efficiency and safety of thymosin-alpha1 treatment in patients with hepatitis B e antigen (HBeAg) and HBV DNA positive chronic hepatitis. METHODS: Sixty-two patients were randomly divided into groups A and B. The patients in group A received subcutaneous injection of 1.6 mg thymosin alpha1, twice a week (T-alpha1 group) for six months, and the patients in group B received 5 MU interferon alpha (IFN-alpha) each day for fifteen days, then three times weekly (IFN-alpha group) for six months. The results between two groups treated with and the group untreated with IFN-alpha which was followed up for 12 mo (historical control group consisting of 30 patients) were compared, and three groups were comparable between each other (P>0.05) at baseline (age, sex, clinical history, biochemical, and serological parameters). RESULTS: At the end of treatment, complete response, which was defined as alanine aminotransferase (ALT) normalization and HBV DNA and HBeAg loss, occurred in 9 of 29 (31.0%) patients in the T-alpha1 group and in 15 of 33 (45.5%) patients in the IFN-alpha group (chi2=1.36, P>0.05). After a follow-up period of six months, a complete response was observed in 14 of 29 (48.3%) patients in the T-alpha1 group and in 9 of 33 (27.3%) patients in the IFN-alpha group (chi2=2.93, P>0.05). Compared with the results observed in the historical control (HC) group untreated with IFN alpha which was followed up for 12 mo, the rate of complete response was significantly higher in IFN-alpha group at the end of therapy (1 of 30 vs 15 of 33, chi2=14.72, P<0.001) and in the T-alpha1 group at the end of follow-up (1 of 30 vs 14 of 29, chi2=15.71, P<0.001). In T-alpha1 and IFN-alpha treatment groups, the area under (the plasma concentration time) curve (AUC) of negative HBV DNA and HBeAg was 34%, 17%, 31% and 19% smaller than that in the HC group. By the end of the follow-up period, the proportions of ALT normalization and negative HBV DNA in the T-alpha1 group were significantly higher than those in the IFN-alpha and HC groups. The odds of ALT normalization and negative HBV DNA at the end of the follow-up was three-fold higher in the T-alpha1 group than in the IFN-alpha group. Unlike IFN-alpha, T-alpha1 was well tolerated by all patients, and no side effects appeared in T-alpha1 group. CONCLUSION: The results suggest that a 6-mo course of T-alpha1 therapy is effective and safe in patients with chronic hepatitis B. T-alpha1 is able to reduce HBV replication in patients with chronic hepatitis B. Furthermore, T-alpha1 is better tolerated than IFN-alpha and can gradually induce more sustained ALT normalization and HBV DNA and HBeAg loss. However, a response rate of 48.3% is still less ideal. A more effective therapeutic approach warrants further study. PMID- 17075993 TI - Hepatocellular carcinoma metastasizing to the skull base involving multiple cranial nerves. AB - We describe a rare case of HCV-related recurrent multiple hepatocellular carcinoma (HCC) metastasizing to the skull base involving multiple cranial nerves in a 50-year-old woman. The patient presented with symptoms of ptosis, fixation of the right eyeball, and left abducens palsy, indicating disturbances of the right oculomotor and trochlear nerves and bilateral abducens nerves. Brain contrast-enhanced computed tomography (CT) revealed an ill-defined mass with abnormal enhancement around the sella turcica. Brain magnetic resonance imaging (MRI) disclosed that the mass involved the clivus, cavernous sinus, and petrous apex. On contrast-enhanced MRI with gadolinium-chelated contrast medium, the mass showed inhomogeneous intermediate enhancement. The diagnosis of metastatic HCC to the skull base was made on the basis of neurological findings and imaging studies including CT and MRI, without histological examinations. Further studies may provide insights into various methods for diagnosing HCC metastasizing to the craniospinal area. PMID- 17075994 TI - Endoscopic management of a new entity-plastobezoar: a case report and review of literature. AB - Foreign bodies and bezoars are commonly encountered in children. We describe a child aged 11 years who ingested large amounts of plastic material used for knitting chairs and charpoys. The conglomerate of plastic threads, entrapped food material and other debris, formed a huge mass occupying the whole stomach. Chronic irritation of the gastric antral mucosa led to ulceration and formation of hyperplastic polyps. We labeled this new entity as a "plastobezoar". The entire bezoar could be removed endoscopically. PMID- 17075995 TI - A case report of hepatic veno-occlusive disease after ingesting dainties. AB - Hepatic veno-occlusive disease (HVOD) is rarely encountered and easily misjudged as Budd-Chiari syndrome. It is often related to stem cell transplantation in recent years. We report a case of HVOD that is related to ingestion of some palatable local dishes. The diagnosis was confirmed by liver biopsy pathology with specific observation of inflammatory changes and fibrosis of venules intima, dilated sinusoids and central veins. Chronic diarrhea is unique for this case as a result of ingesting harmful stuffs. This case demonstrated that supervision and instruction of food recipe and traditional medicine are crucial, and prompt diagnosis, supportive care and specific treatment are essential to decreasing the morbidity and mortality of HVOD. PMID- 17075996 TI - [Pharmacologic interest in HIV protease inhibitors: concepts and utilization]. PMID- 17075997 TI - [Resistance to protease inhibitors: the burden of mutations]. PMID- 17075998 TI - [Long term virologic efficacy of antiretroviral agents]. PMID- 17075999 TI - [Lipid profile for treatment with protease inhibitors]. PMID- 17076000 TI - [Co-infection with HIV and viral hepatitis C]. PMID- 17076001 TI - [New therapeutic strategies. Many of the trails have not yet been validated]. PMID- 17076002 TI - [Malignant hemopathies and tumor lysis syndrome: a current topic]. PMID- 17076003 TI - [The physiological basis of the treatment of tumor lysis syndrome]. PMID- 17076004 TI - [The current status of the treatment of tumor lysis syndrome--the role of rasburicase (Fasturtec)]. PMID- 17076006 TI - Medscape. PMID- 17076005 TI - [Recommendations of the French Society for the Control of Cancers and Leukemias in Children for the treatment of tumor lysis syndrome: results of a pediatric survey]. PMID- 17076007 TI - Delivery of 100%, 40%, and blow-by humidity did not differ for change in croup scores in children with moderate croup. PMID- 17076008 TI - A self directed behavioural family intervention with telephone consultations improved toddler behaviour and parenting skills. PMID- 17076009 TI - Review: vaccines prevent influenza and influenza-like illness in healthy children <16 years of age. PMID- 17076010 TI - Review: antiviral agents reduce risk of influenza in healthy adults and alleviate symptoms faster than placebo. PMID- 17076011 TI - Review: existing evidence does not support behavioural or other specific interventions to prevent smoking relapse. PMID- 17076012 TI - Review: self testing and self management increase the benefits and reduce the harms of anticoagulant therapy. PMID- 17076013 TI - A low fat dietary pattern intervention did not reduce breast cancer, colorectal cancer, or CVD in postmenopausal women. PMID- 17076014 TI - Calcium plus vitamin D did not prevent fractures or colorectal cancer in postmenopausal women. PMID- 17076015 TI - Allowing wounds to be uncovered and wet in the first 48 hours after minor skin excision did not differ from standard dry management for wound infections. PMID- 17076016 TI - Review: full body washing with skin antiseptics before surgery does not prevent surgical site infection. PMID- 17076017 TI - A target backrest elevation of 45 degrees was not feasible for mechanically ventilated patients; elevations achieved did not prevent ventilator associated pneumonia. PMID- 17076019 TI - Maintenance treatment with paroxetine, but not psychotherapy, prevented recurrent major depression in older people. PMID- 17076018 TI - Review: behaviour therapy is effective for insomnia. PMID- 17076020 TI - Review: self report of hearing loss and the whispered voice test are useful for screening for hearing impairment. PMID- 17076021 TI - Physical exertion at work during pregnancy did not increase risk of preterm delivery or fetal growth restriction. PMID- 17076022 TI - Training and support for nursing home staff reduced neuroleptic drug use and did not increase aggression in residents with dementia. PMID- 17076023 TI - Women offered prenatal screening had varying responses on becoming candidates for higher risk status. PMID- 17076024 TI - Mothering served as a narrative backbone that supported a meaningful, well developed life in some teenage mothers. PMID- 17076025 TI - Review: meta-analysis of qualitative studies generated recommendations for healthcare professionals meeting with women who had experienced intimate partner violence. PMID- 17076026 TI - The masculine concepts of control, strength, and responsibility to others were important to men with depression. PMID- 17076027 TI - Delays in the diagnosis of lung cancer occurred because patients failed to recognise symptoms as serious and warranting medical attention. PMID- 17076028 TI - "Packaging" information about patient deterioration in terms of vital signs and the Early Warning Score facilitated nurses' communication with doctors. PMID- 17076029 TI - Pulpal response to a newly developed MMA based resin cement for bonding tooth colored indirect restorations. AB - PURPOSE: To evaluate the pulpal responses to a newly-developed MMA-based self etch resin cement, when used as a luting agent for indirect resin composite restoration, and to compare the results with those obtained from a total-etch luting agent, glass-ionomer cement, and amalgam restoration. METHODS: 120 cervical cavities were prepared in monkey teeth and divided into four equal groups according to the restorative materials used: (1) the cavities were restored with resin composite inlays using a self-etch resin cement as a luting agent (M-Bond); (2) the cavities were also restored with resin composite inlays but using a total-etch resin cement as a luting agent (Super-Bond C & B); (3) the cavities were directly restored with glass-ionomer cement (Fuji II); or (4) the cavities were directly restored with amalgam (Dispersalloy). The restored teeth were extracted at 3, 30, or 90 days after restoration, then fixed in 10% neutral buffered formalin. The specimens were prepared using routine histopathological procedures. Five microm-thick sections were stained with hematoxylin and eosin or Brown & Brenn gram stain for bacterial observations. Histological responses in the pulpal tissue and bacterial penetration were observed under a light microscope and evaluated using standard scores. The results were statistically analyzed using the Kruskal-Wallis test (P< 0.05). RESULTS: At all time intervals, no significant differences of pulpal inflammatory responses between M-Bond and Super-Bond C&B were observed (P> 0.05). Both resin cements showed no serious pulpal responses, such as necrosis or abscess formation. In general, both MMA based resin cements showed similar pulpal responses to those of glass-ionomer cement except for congestion of pulpal blood vessels at 3 days after restoration in which glass-ionomer cement exhibited a lower level than that of the MMA-based resin cements. For the group restored with amalgam, at 3 days after restoration, severe odontoblastic disorders and blood vessel congestions with a large infiltration of inflammatory cells were detected. At 30 and 90 days after restoration, slightly inflammatory irritations were observed irrespective of the materials used. Reparative dentin formation and bacterial penetration were found mostly in the group restored with amalgam. PMID- 17076030 TI - Final report and recommendations from the National Notifiable Diseases Working Group. AB - The National Notifiable Diseases Working Group performed a ranking of 48 communicable diseases to assist with determining priorities for national surveillance. The WG offers six specific recommendations relating to the addition or deletion of communicable diseases from the list of nationally notifiable diseases. KEY RECOMMENDATIONS: 1. Clostridium difficile-associated diarrhea, paralytic shellfish poisoning, and invasive listeriosis are recommended for addition to the Nationally Notifiable Disease List. 2. New proposals for diseases not recommended for national notification include hepatitis E, HTLV-1, and HTLV 2. 3. Dengue virus infection should be deleted from the Nationally Notifiable Disease List. (Dengue hemorrhagic fever should continue to be reported.) 4. Classic Creutzfeld-Jacob disease ranked well below the threshold for inclusion; however, it should be retained on the list because surveillance of classic CJD is key to effective surveillance of new variant disease and because consulted provinces and territories strongly favoured its retention. 5. Several diseases that ranked near the borderline for reporting should be retained on the list as follows: a. a. brucellosis, because although it does not rank highly on the basis of its endemic pattern it is a category B biological warfare agent; b. laboratory confirmed influenza because it forms part of a functional and working surveillance approach; c group B streptococcal infection because alternative hospital-based surveillance systems do not yet capture a significant proportion fo cases. 6. Transfusion-transmitted infections were unrankable by the current system. 7. The approach to reporting dengue hemorrhagic fever, plague, West Nile virus infections, and other viral hemorrhagic fevers should be made congruent with the approach for other notifiable diseases (see Discussion). 8. The feasibility and utility of national reporting for communicable diseases that have very high incidence and low severity (e.g. chickenpox, norovirus) should be carefully considered, especially when it leads to little or no case-by-case data reported by health authorities. The WG recommends that these diseases be placed under surveillance but that case-by-case reporting not be required at the national level. Alternative approaches to tracking trends could include enumerating outbreaks in the case of norovirus or the use of sentinel surveillance, laboratory surveillance, or physician billing events in the case of varicella. PMID- 17076031 TI - Outbreak of tuberculosis in the Laurentian region, 2005. PMID- 17076032 TI - Believing in brain and brawn. PMID- 17076033 TI - Home or away? Investigation of Salmonella enteritidis PFGE pattern SENXAI.0003 and SENBNI.0003, phage type 8, the in the Maritimes, 2005. PMID- 17076034 TI - The changing epidemiology of hepatitis A in British Columbia: using health authority follow-up data to inform policy and practice. PMID- 17076035 TI - Microfluidic proteomic reactor. PMID- 17076036 TI - Age-related macular degeneration and the RPE secretome. PMID- 17076037 TI - NCI CPTI: technologies for proteomics. PMID- 17076038 TI - A successful partnership to improve prescribing practices. PMID- 17076039 TI - [Bozena Grabda-Kazubska (1936-1999)]. PMID- 17076041 TI - Considerations of the institutional review board in artificial organ development. PMID- 17076040 TI - Morgentaler v. The Queen in the Supreme Court of Canada. AB - In Morgentaler v. R., the Supreme Court of Canada struck down the abortion provisions in the Criminal Code. In a five to two split, a majority of the Supreme Court judges found that section 251 offended a pregnant woman's constitutionally protected right not to be deprived of her "life, liberty, and security of the person." Sheilah Martin reviews the three majority judgments and focuses on the decision written by Madame Justice Wilson. She believes that Madame Justice Wilson's opinion merits special attention in several regards: her conclusions on the constitutional rights of pregnant women; her recognition and validation of women's perspectives on abortion; and her approach to balancing women's interests in reproductive self-determination against the state's interest in regulating reproduction. Sheilah Martin concludes that this decision will reverberate far into the future. Even though it fails to establish clear guidelines concerning governmental power to control access to abortion, its principles outline the legal framework in which future litigation will occur, and it will limit and shape the terms of any ensuing political debate. In addition, Madame Justice Wilson's judgment holds great promise for those looking to the Court to promote the rights of women and other historically disadvantaged groups. PMID- 17076042 TI - Detrimental therapist-client relationships--beyond thinking of "dual" or "multiple" roles: reflections on the 2001 AAMFT Code of Ethics. AB - This article presents reflections on and a critique of the recent revision of the AAMFT Code of Ethics on the multiple relationship ethical standard. A brief historical overview of terminology and the debate surrounding "dual" and "multiple" relationship ethical rules in marriage and family therapy is provided. The term "exploitation" is also delimited. Ethical principles and a set of standards addressing "detrimental" versus "potentially beneficial" interactions are introduced, deriving from works in other mental health professions. The article recommends: (a) the terms "dual" and "multiple" relationships should be abandoned; (b) the ethical principles underlying the AAMFT Code of Ethics need to be examined; and (c) the debate on the topic of detrimental therapist-client interactions in marriage and family therapy needs to be revisited, especially in light of a "positive ethics." PMID- 17076043 TI - 'Four legs good, two legs bad'--political spin in the HIV prevalence survey. PMID- 17076044 TI - Cardiac catheterization. PMID- 17076045 TI - Cardiovascular perfusionist. PMID- 17076046 TI - Physician assistant. PMID- 17076047 TI - Electroneurodiagnostic technologist. PMID- 17076048 TI - In re Doe. PMID- 17076049 TI - Medicare Part B income-related monthly adjustment amount. Final rules. AB - We are adding to our regulations a new subpart, Medicare Part B Income-Related Monthly Adjustment Amount, to contain the rules we will follow for Medicare Part B income-related monthly adjustment amount determinations. The monthly adjustment amount represents the amount of decrease in the Medicare Part B premium subsidy, i.e. the amount of the Federal Government's contribution to the Federal Supplementary Medical Insurance (SMI) Trust Fund. This new subpart implements section 811 of the Medicare Prescription Drug, Improvement, and Modernization Act of 2003 (the Medicare Modernization Act or MMA) and contains the rules for determining when, based on income, a monthly adjustment amount will be added to a Medicare Part B beneficiary's standard monthly premium. These final rules describe: What the new subpart is about; what information we will use to determine whether you will pay an income-related monthly adjustment amount and the amount of the adjustment when applicable; when we will consider a major life changing event that results in a significant reduction in your modified adjusted gross income; and how you can appeal our determination about your income-related monthly adjustment amount. PMID- 17076050 TI - System transformation through best practices. PMID- 17076051 TI - A six-month outcome of long-stay inpatients resettled in a hostel. AB - This prospective study evaluated a 6-month outcome of resettlement of long-term psychiatric inpatients to a hostel-based residence. Participants included 92 patients with DSM-IV chronic psychotic disorders, aged 18 to 65 years, hospitalized for 16.7 +/- 10.6 years. Evaluations performed at 2 weeks before their resettlement and following 6 months of hostel residence included the Positive and Negative Syndromes Scale, Montgomery and Asberg Depression Scale, Clinical Global Impression, Global Assessment of Functioning, Social Adaptive Functioning Evaluation and Social Behavior Schedule. The Nurses' Observation Scale for Inpatient Evaluation was completed regularly. Psychopathological symptoms declined and social interaction increased. Hostel-based deinstitutionalization programs are a promising alternative for long-term psychiatric inpatients. PMID- 17076052 TI - The Recovery Knowledge Inventory: assessment of mental health staff knowledge and attitudes about recovery. AB - In order to assess knowledge of and attitudes toward recovery-oriented practices among providers of mental health and substance abuse treatment and rehabilitation services throughout the state of Connecticut, an instrument named the Recovery Knowledge Inventory (RKI) was developed and administered. The items that comprise the instrument are based on the emerging literature on recovery in psychiatric and substance use disorders, and assess four different domains of understanding, namely: 1) roles and responsibilities in recovery, 2) non-linearity of the recovery process, 3) the roles of self-definition and peers in recovery, and 4) expectations regarding recovery. This paper describes the instrument and its preliminary psychometric properties, and provides an example of its utility in assessing the training needs of staff who increasingly are being expected to deliver recovery-oriented care. PMID- 17076053 TI - A group intervention to promote healthy self-concepts and guide recovery in first episode schizophrenia: A pilot study. AB - The stress associated with managing a first episode of schizophrenia and the secondary psychological sequelae may predispose young people to the phenomenon of illness engulfment, whereby personal identity is lost and replaced with a sense of self defined entirely by the illness. The overall objective of this pilot project was to provide an initial evaluation of the impact of a novel group intervention targeting improvement of self-concept (engulfment) and overall quality of life for young adults recovering from a first episode of schizophrenia. Fifty-two young adults diagnosed with a DSM-IV schizophrenia spectrum disorder were sequentially assigned to either a 12-week group intervention that provided members with opportunities to attain healthy self concepts or a control group that received high quality treatment as usual. The two groups were compared on pretreatment, post-treatment, and 3-month post treatment measures of engulfment, quality of life, and psychiatric symptoms. Allowing for dropouts, 26 subjects remained in the treatment group and 14 subjects in the control group after the 3-month follow-up. Participants in the treatment group demonstrated significant improvement in engulfment, quality of life (intrapsychic subscale) and symptoms, while the comparison group did not change. These preliminary results provide support for the benefits of a group intervention designed to enhance self-concept and to minimize the engulfing effects of illness. PMID- 17076054 TI - ACT team members' responses to training in recovery-oriented practices. AB - To further our understanding of the consequences of training in recovery principles and practices, this study examined ACT team workers' responses to a state-wide recovery training initiative. Analysis of trainees' comments revealed ten themes expressing endorsement of or difficulties with recovery-oriented practices. Trainees' comments supporting a recovery orientation described service recipients in holistic terms and described using various techniques to attain "client-centered" goals. Recovery-oriented tasks that posed dilemmas for trainees were: reconciling system-centered goals with recipients' goals, establishing collaborative relationships with recipients, and using a recovery orientation with recipients who are in crisis and/or who "don't admit to being mentally ill." PMID- 17076055 TI - Prediction of work rehabilitation placements using the Chinese Work Personality Profile. AB - This study aims to evaluate the application of a situational assessment instrument, the Chinese Work Personality Profile (CWPP), to predict the work rehabilitation placements of persons with psychiatric disabilities. Using the five CWPP subscale scores as predictors, both discriminant and classification and regression tree analysis showed that the CWPP scores correctly identified more than 80% of the work rehabilitation placements of the 179 participants. Social skills, self-control, and task orientation factors of the CWPP were the most important predictors of work rehabilitation placements, and clinical implications for these results were discussed. PMID- 17076056 TI - Collaborative goal technology: theory and practice. AB - Goal striving promotes hope and enhances motivation, which is important for psychosocial rehabilitation and recovery. The Collaborative Goal Technology (CGT) is a new goal striving intervention that is used to support the autonomy and recovery processes of the person with a psychiatric disability. The CGT protocol and its utility are outlined. Theory and research from goal striving, motivation and mental health recovery domains that informed the development of CGT are described. A case example is also provided. PMID- 17076057 TI - Employing behavioral methods to improve the context of care within a public psychiatric hospital: enhancing psychosocial programming. AB - This article reviews efforts, grounded in principles of behavioral science, to improve the "goodness of fit" between the needs of psychiatrically impaired individuals and the array of day program classes offered in a public psychiatric hospital. The relevance of behavioral science and technology for planning and implementing such change in resource-challenged organizations is discussed. PMID- 17076058 TI - Implementing program-wide awareness about recovery in a large mental health and addictions program. AB - St. Joseph's Healthcare Hamilton conceptualized a multi-step plan for implementing a recovery oriented service delivery approach within their Mental Health and Addictions Program. This brief report discusses the first phase of this plan which included building awareness of recovery utilizing Anthony's (2000) system standards to develop a needs assessment for managers and senior team members. The survey had three purposes: to increase managers' awareness about recovery; to allow managers to express concerns that they had with this paradigm; and to afford managers an opportunity to explore the ways in which their service was and was not operating in a recovery oriented way. Initiatives designed to build awareness throughout the program are discussed. PMID- 17076059 TI - Experience with post-secondary education for individuals with severe mental illness. AB - There is a need for increased understanding and support of students with severe and persistent psychiatric disabilities in post-secondary education. Advancements in the pharmacological treatment of psychiatric disabilities are permitting a broader range of mental health consumers to reach a level of recovery allowing these consumers to begin or restart college careers (Eudaly, 2002). A post secondary education is now considered a mandatory investment in the future ("Higher Education," 2003). There are a growing number of people pursuing higher education including those with serious mental illness. I am working on a Master's degree in a Rehabilitation Counselor Education program and face many obstacles due to my psychiatric disability, Bipolar Disorder. PMID- 17076060 TI - In-service training: making it stick. PMID- 17076061 TI - Flagging early examples of ambiguity II. PMID- 17076062 TI - Effects of motion parallax and perspective cues on perceived size and distance. AB - In three experiments we examined the relative effectiveness of motion parallax and two perspective cues for the perception of size and distance. The experimental stimuli consisted of two ellipses (a standard and a comparison) and a horizontal line that indicated the horizon. The subject's task was to report the apparent size and distance of the comparison stimulus relative to the standard stimulus. Two perspective cues were given by the relative heights of the two stimuli and the absolute height of the standard stimulus below the horizon. Motion parallax was defined by both the ratio and the difference in angular velocities between the two stimuli on the display. In experiment 1 we examined the effects of the two perspective cues and the motion parallax. In experiment 2 we eliminated the horizon line, and examined the role of the horizon in size and distance perception. In experiment 3 we separately evaluated the effects of motion parallax and the relative height cues. The results from the three experiments showed that the effect of motion parallax and the two perspective cues were different in three ways. First, the relative effectiveness of motion parallax and the two perspective cues differed for size and distance estimates. For size estimates, the motion parallax was more effective than the perspective cues (experiments 1 and 3). For distance estimates, the motion parallax was as effective as the two perspective cues (experiments 1 and 3). Second, the role of the horizon differed for size and distance estimates. The size estimates were strongly affected by the horizon, while the distance estimates were not affected much by the horizon (experiment 2). Third, the effective perspective cues differed for the size estimates and the distance estimates: size estimates were affected by the perspective cues as a combination of the horizon and relative height; distance estimates were affected by the perspective cues as an interaction between the absolute and relative heights without the horizon line. PMID- 17076063 TI - Synaesthesia: the prevalence of atypical cross-modal experiences. AB - Sensory and cognitive mechanisms allow stimuli to be perceived with properties relating to sight, sound, touch, etc, and ensure, for example, that visual properties are perceived as visual experiences, rather than sounds, tastes, smells, etc. Theories of normal development can be informed by cases where this modularity breaks down, in a condition known as synaesthesia. Conventional wisdom has held that this occurs extremely rarely (0.05% of births) and affects women more than men. Here we present the first test of synaesthesia prevalence with sampling that does not rely on self-referral, and which uses objective tests to establish genuineness. We show that (a) the prevalence of synaesthesia is 88 times higher than previously assumed, (b) the most common variant is coloured days, (c) the most studied variant (grapheme-colour synaesthesia)--previously believed most common--is prevalent at 1%, and (d) there is no strong asymmetry in the distribution of synaesthesia across the sexes. Hence, we suggest that female biases reported earlier likely arose from (or were exaggerated by) sex differences in self-disclosure. PMID- 17076064 TI - Direct evidence for the existence of energy-based texture mechanisms. AB - Two classes of models have been proposed to explain how the visual system processes texture modulations. In 'feature models', abstract representations of the featural properties of local texture regions (eg orientation, spatial frequency, contrast) are first generated, after which differences in individual feature properties across space are detected. In 'energy models', on the other hand, differences across space in the response energies of linear simple-cell like filters are detected. This model thus processes the existing differences between texture regions directly without generating a full representation of the individual texture regions. We provide here direct evidence for the existence of the second, energy model, using an adaptation paradigm in conjunction with textures simultaneously modulated in two dimensions--orientation and spatial frequency. We found that the mechanism that processed the conjoint modulation was tuned to orientations and spatial frequencies that could not be predicted by any feature model, but which were precisely predicted by the energy model. PMID- 17076065 TI - Visual form-processing deficits in autism. AB - People with autism have a number of reported deficits in object recognition and global processing. Is there a low-level spatial integration deficit associated with this? We measured spatial-form-coherence detection thresholds using a Glass stimulus in a field of random dots, and compared performance to a similar motion coherence task. A coherent visual patch was depicted by dots separated by a rotational transformation in space (form) or space-time (motion). To measure parallel visual integration, stimuli were presented for only 250 ms. We compared detection thresholds for children with autism, children with Asperger syndrome, and a matched control group. Children with autism showed a significant form coherence deficit and a significant motion-coherence deficit, while the performance of the children with Asperger syndrome did not differ significantly from that of controls on either task. PMID- 17076066 TI - Units of visual individuation in rhesus macaques: objects or unbound features? AB - Vision begins with the processing of unbound visual features, which must eventually be bound together into object representations. Such feature binding is required for coherent visual perception, and accordingly has received a considerable amount of study in several domains. Neurophysiological work, often in monkeys, has revealed the details of how and where feature binding occurs in the brain, but methodological limitations have not allowed this research to elucidate just how feature binding operates spontaneously in real-world situations. In contrast, behavioral work with human infants has demonstrated how we use simpler unbound features to individuate and identify objects over time and occlusion in many types of events, but this work has not typically been able to isolate the role of feature binding in such processing. Here we provide a method for assessing the spontaneity and fidelity of feature binding in non-human primates, as this process is utilized in real-world situations, including simple foraging behaviors. Using both looking-time and manual-search measures in a natural environment, we show that free-ranging rhesus macaques (Macaca mulatta) spontaneously bind features in order to individuate objects across time and occlusion in dynamic events. This pattern of results demonstrates that feature binding is used in subtle ways to guide ecologically relevant behavior in a non human animal, spontaneously and reliably, in its natural environment. PMID- 17076067 TI - The role of part structure in the perceptual localization of a shape. AB - The process of object localization may be accomplished with respect to a particular reference location, such as the center of gravity, COG (eg Vishwanath and Kowler, 2003 Vision Research 43 1637-1653). Here, we investigated how part structure affects an object's reference location. The reference location was evaluated with a measure of the illusory displacement of an internal target element embedded within a larger object (Morgan et al, 1990 Vision Research 30 1793-1810). To examine whether the reference location is different for shapes with part structure, two shapes were tested: circle (small and large; no part structure) and bell (shape with two parts, one larger than the other). Results were examined with respect to two predictions: either the location of an object is based on its shape as a whole, disregarding part structure (ie a single, overall COG), or the parts are processed separately (different COGs). With the circles, the results showed a systematic illusory displacement of the internal target toward the COG. With the bell, the illusion was significantly weaker than with both circles--even though the main part of the bell had the same size as the small circle, and its horizontal axis had the same extent as the large circle. Moreover, the distance judgments for the bell were consistent with a (weaker) reference point being located at the COG of the larger part, rather than at the COG of the entire bell. These results show that the part structure of a shape plays a role in the representation of its location, and that for complex shapes the perceived location of an embedded element depends more on the parts within which it is embedded, rather than on the whole shape. PMID- 17076068 TI - Information processing during face recognition: the effects of familiarity, inversion, and morphing on scanning fixations. AB - Where we make ocular fixations when viewing an object likely reflects interactions between 'external' object properties and internal 'top-down' factors, as our perceptual system tests hypotheses and attempts to make decisions about our environment. These scanning fixation patterns can tell us how and where the visual system gathers information critical to specific tasks. We determined the effects of the internal factors of expertise, experience, and ambiguity on scanning during a face-recognition task, in eight subjects. To assess the effects of expertise, we compared upright with inverted faces, since it is hypothesized that inverted faces do not access an orientation-dependent face-expert processor. To assess the effects of experience, we compared famous with novel faces, as famous faces would have stronger internal representations than anonymous ones. Ambiguity in matching seen and remembered faces was manipulated with morphed faces. We measured three classes of variables: (i) total scanning time and fixations; (ii) the spatial distribution of scanning; and (iii) the sequence of scanning, using first-order Markov matrices for local scan structure and string editing for global scan structure. We found that, with inverted faces, subjects redistributed fixations to the mouth and lower face, and their local and global scan structure became more random. With novel or morphed faces, they scanned the eyes and upper face more. Local scan structure was not affected by familiarity, but global scan structure was least random (most stereotyped) for novel upright faces. We conclude that expertise (upright faces) leads to less lower-face scanning and more predictable global patterns of information gathering. Experience (famous faces) leads to less upper-face scanning and more idiosyncratic global scan structures, suggesting a superseding influence of facial memories. With morphed faces, subjects return to the upper face to resolve ambiguity, implying a greater importance of this region in face recognition. PMID- 17076069 TI - Featural and configural face processing in adults and infants: a behavioral and electrophysiological investigation. AB - We sought to elucidate the behavioral and electrophysiological correlates of face processing, in adults and infants, by manipulating either the featural or configural information within the face. Two different experiments are reported. In these experiments, event-related potentials (ERPs) were recorded from the scalp while adult, 8-month-old, and 4-month-old participants completed configural change and featural-change face tasks. The infants also completed a behavioral visual paired-comparison task with featural and configural face changes. ERP results reveal hemispheric differences in processing featural but not configural changes for the N170 in adults. Furthermore, featural and configural changes are processed differently within the right and left hemispheres. The right hemisphere N170 is significantly greater for configural compared to featural changes. The left hemisphere N170, however, exhibits the opposite effect. Infant data suggest that similar to adults, 8-month-old, but not 4-month-old participants, exhibit similar hemispheric differences between featural and configural changes for the P400 component. Behavioral results suggest increased sensitivity to both featural and configural face changes in 8-month-olds compared to 4-month-olds. PMID- 17076070 TI - Females, but not males, show greater pupillary response to direct- than deviated gaze faces. AB - Under suitable conditions, pupillary dilation is a reliable index of processing activity. Pupil size was tracked in male and female observers following the presentation of face stimuli for an age-judgment task. The eyes on the faces were either directed towards the observer or deviated to the side. Pupil dilation accompanied processing of the faces, but female observers showed significantly more sustained pupil dilation when viewing direct- than deviated-gaze faces over the period 3 to 7 s after stimulus onset, regardless of stimulus sex. In contrast, male observers did not show a consistent pattern in response to either the gaze or sex of the face stimuli. These findings indicate a sex difference in the processing of gaze direction and suggest that females, but not males, apply increased effort to processing socially relevant (direct-gaze) than irrelevant (deviated-gaze) faces. They also demonstrate that pupillary measurement can potentially provide new insights into the processing of even visual input, provided reflexes are sufficiently controlled. PMID- 17076072 TI - California, here we come. PMID- 17076071 TI - Perception of elliptic biological motion. AB - We tested the ability of the mature visual system for discrimination between types of elliptic biological motion on the basis of event kinematics. Healthy adult volunteers were presented with point-light displays depicting elliptic motion when only a single dot, a moving point-light arm, or a whole point-light human figure was visible. The displays were created in accordance with the two thirds power kinematic law (natural motion), whereas the control displays violated this principle (unnatural motion). On each trial, participants judged whether the display represented natural or unnatural motion. The findings indicate that adults are highly sensitive to violation of the two-thirds power kinematic law. Notably, participants can easily discriminate between natural and unnatural motions without recognising the stimuli, which suggests that people implicitly use kinematic information. Most intriguing, event recognition seems to diminish the capacity to judge whether event kinematics is unnatural. We discuss possible ways for a cross-talk between perception and production of biological movement, and the brain mechanisms involved in biological motion processing. PMID- 17076073 TI - Blastomere blowup. A novel way to harvest stem cells intrigues and inflames. PMID- 17076074 TI - Keeping CO2 down. PMID- 17076075 TI - Muscling up color. PMID- 17076076 TI - Catchy carbon. What Floyd Landis has in common with ocean sediment. PMID- 17076077 TI - Martian field test. Drilling in Greenland to prepare for Mars. PMID- 17076078 TI - Not imagining it. Research into hallucinogens cautiously resumes. PMID- 17076079 TI - Myth: red-blue states. American split in culture war unfounded. PMID- 17076080 TI - Wronger than wrong. Not all wrong theories are equal. PMID- 17076081 TI - Welfare states, beyond ideology. PMID- 17076083 TI - The dark ages of the universe. PMID- 17076082 TI - The evolution of future wealth. PMID- 17076084 TI - Mirrors of the mind. PMID- 17076085 TI - Broken mirrors: a theory of autism. PMID- 17076086 TI - Malware goes mobile. PMID- 17076087 TI - Reviving dead zones. PMID- 17076088 TI - Seeing with superconductors. PMID- 17076090 TI - Gunk-free fiber. PMID- 17076089 TI - The origin of the Greek constellations. PMID- 17076091 TI - Does obstructive sleep apnea affect aerobic fitness? AB - OBJECTIVES: We sought to determine whether patients with obstructive sleep apnea (OSA) had an objective change in aerobic fitness during cycle ergometry compared to a normal population. The most accurate test of aerobic fitness is measurement of maximum oxygen consumption (VO2max) with cycle ergometry. METHODS: We performed a retrospective cohort analysis (247 patients with OSA) of VO2max from annual cycle ergometry tests compared to a large control group (normative data from 1.4 million US Air Force tests) in a tertiary care setting. RESULTS: Overall, individuals with OSA had increased VO2max when compared to the normalized US Air Force data (p < .001). Patients with an apnea-hypopnea index of greater than 20 demonstrated a decreased VO2max as compared to normalized values (p < .001). No differences in VO2max were observed after either medical or surgical therapy for OSA. CONCLUSIONS: Overall, in a US Air Force population, OSA does not predict a decrease in aerobic fitness as measured by cycle ergometry. However, patients with an apnea-hypopnea index of greater than 20 have a statistically significant decrease in aerobic fitness compared to the normal population. This study demonstrates the effects of OSA on aerobic fitness. Further correlation of fitness testing results with OSA severity and treatment is needed. PMID- 17076092 TI - Correlation of radiologic findings and clinical outcome in pharyngeal pouch stapling. AB - OBJECTIVES: We undertook to identify any correlation between radiologic findings and clinical outcome in pharyngeal pouch surgery. METHODS: We performed a retrospective case control study in a teaching hospital in Dundee, Scotland. The subjects were 21 patients who underwent pharyngeal pouch stapling. We used a novel method to measure the characteristics of each pouch by focusing on a triangular area in the neck of the pouch. RESULTS: A correlation was found between the radiologic preoperative findings and the clinical outcome. A short triangle was more likely to result in postoperative complications, whereas a long one was more likely to require revision surgery. CONCLUSIONS: It is possible to predict the surgical outcome from the radiologic characteristics of the diverticulum. We believe that our findings may have important clinical implications. PMID- 17076093 TI - Advantage of accelerated fractionation regimens in definitive radiotherapy for stage II glottic carcinoma. AB - OBJECTIVES: We evaluated the prognostic factors for local control of T2 glottic cancer and verified the efficacy of accelerated fractionation regimens such as hyperfractionation and accelerated hyperfractionation. METHODS: A total of 86 patients with T2 N0 M0 glottic squamous cell carcinoma, who were treated with definitive radiotherapy, were analyzed retrospectively by multivariate analysis. RESULTS: Overall treatment time of radiotherapy (p = .0003) and total dose (p = .0036) were the significant prognostic factors for local control on multivariate analysis. The group with a higher total dose (> or = 67 Gy versus <67 Gy) showed a favorable prognosis (5-year local control rate of 91% versus 60%, respectively; p = .0013, log-rank test). Likewise, the group with a shorter overall treatment time of radiotherapy (< or = 54 days versus >54 days) showed a favorable prognosis (5-year local control rate of 87% versus 71%, respectively; p = .023). CONCLUSIONS: A radiotherapy total dose of > or = 67 Gy delivered for a shorter period is required for T2 glottic cancer. The fractionation regimens of hyperfractionation and accelerated hyperfractionation are more effective than conventional fractionation in terms of shortening overall treatment time and delivering a high total dose with acceptable toxicity. PMID- 17076094 TI - Comparison of a flexible laryngoscope with calibrated sizing function to intraoperative measurements. AB - OBJECTIVES: The objectives were to assess the clinical performance and accuracy of a prototype fiberoptic transnasal laryngeal endoscope with an auxiliary optical system that allows images to be spatially calibrated. METHODS: A novel fiberoptic endoscope was developed that projects green laser beams across the field of view from a separate optical channel. According to the location of the spots in the field of view, the images can be calibrated with a software routine. To assess its performance, we compared measurements of 14 lesions imaged with the calibrated endoscope and during microlaryngoscopy, where a calibration instrument was placed next to the lesions. Four clinicians measured lesion length, width, and area from the collected images. RESULTS: The calibrated endoscope performed as well as current flexible fiberoptic laryngoscopes in terms of image quality and patient comfort. For lesions with well-defined borders, the error ranged from 14% to 23% for length, from 20% to 30% for width, and from 33% to 50% for area across observers. Factors contributing to larger errors in some subjects were identified. CONCLUSIONS: The calibrated endoscope is capable of providing useful sizing information for laryngeal structures, and these measures correspond quite well to more direct measurements in the operating room. Objective sizing of laryngeal lesions is complicated by subjective judgments of lesion boundaries, which can be indistinct in many cases. PMID- 17076095 TI - Expression and distribution of CD44 and hyaluronic acid in human vocal fold mucosa. AB - OBJECTIVES: Expression of CD44 (a cell surface receptor for hyaluronic acid) and the distribution of hyaluronic acid were examined in the human vocal fold mucosa. METHODS: Light microscopic investigation was carried out on 10 normal larynges of newborn, infant, younger adult, and older adults with Alcian blue staining, a hyaluronidase digestion study, and immunohistochemistry for CD44. RESULTS: Before the appearance of hyaluronic acid in the newborn vocal fold mucosa, CD44 was expressed on the stellate cells in the macula flava and on the fibroblasts in Reinke's space. During infancy, hyaluronic acid appeared and was distributed in the vocal fold mucosa. Many more stellate cells in the macula flava showed CD44 expression, and a large amount of hyaluronic acid was present around the infant stellate cells; however, the fibroblasts in Reinke's space expressed little CD44. During adulthood, hyaluronic acid was distributed in the vocal fold mucosa. Almost all of the stellate cells in the macula flava showed CD44 expression, and much hyaluronic acid existed around the stellate cells in the adult macula flava. However, fibroblasts in the adult Reinke's space expressed little CD44, and hyaluronic acid density in that space was lower than that in the macula flava. CONCLUSIONS: Stellate cells in the macula flava and CD44 cooperatively play important roles in maintaining hyaluronic acid in the human vocal fold mucosa as a vibrating tissue. PMID- 17076096 TI - Evaluation of recovery from transient facial palsy following canalplasty and tympanoplasty for the treatment of congenital aural atresia. AB - OBJECTIVES: Canalplasty is a surgical procedure of the external auditory canal. In this study we examined anatomic risk factors for facial nerve injury in aural atresia surgery, in addition to facial nerve outcomes and the time course of recovery in patients with facial nerve palsy following atresia surgery. METHODS: Transient facial palsy was observed immediately after surgery in 6 of 99 ears (6 of 87 patients) after canalplasty and tympanoplasty for the treatment of congenital aural atresia. We assessed the course of recovery and mean scores using Jahrsdoerfer's grading system. RESULTS: The rate of absence of the stapes and the rate of presence of positional anomalies of the facial nerve were significantly higher among cases with facial palsy, and Jahrsdoerfer total scores were significantly lower among cases with facial palsy. The mean recovery time in 5 cases with a minimum electroneurography value of 0% was 16.2 weeks (range, 14 to 18 weeks). No trends during recovery were detected for the nerve excitability test and R1 components of the blink reflex. CONCLUSIONS: The anatomic conditions that cause a predisposition to transient facial palsy include absence of the stapes and the presence of positional anomalies of the facial nerve. Although some patients displayed transient facial palsy after surgery, all patients fully recovered. Mechanisms of facial nerve injury are proposed. PMID- 17076097 TI - Respiratory retraining therapy and management of laryngopharyngeal reflux in the treatment of patients with cough and paradoxical vocal fold movement disorder. AB - OBJECTIVES: We describe the outcome of patients with cough and paradoxical vocal fold movement disorder (PVFMD) treated with respiratory retraining therapy and management of laryngopharyngeal reflux (LPR). METHODS: Twenty patients with the complaint of cough were given a diagnosis of PVFMD and treated with proton pump inhibitors for a minimum of 6 months followed by 3 to 5 sessions of respiratory retraining therapy. Pulmonary function testing (PFT) and subjective rating of cough and reflux (reflux symptom index; RSI) were performed. Also, PFT and rating of cough were performed on a group of 10 healthy volunteers with no complaint of cough. RESULTS: The study group comprised 13 women and 7 men. The baseline cough rating and ratio of forced inspiratory volume at 0.5 second to forced inspiratory vital capacity (FIV0.5/FIVC) on PFT were significantly worse in the treatment group than in the control group. After therapy, 20 patients (100%) experienced improvement in cough, 19 patients (95%) experienced improvement on PFT, and 17 patients (85%) experienced improvement in the RSI score. The differences were statistically significant. CONCLUSIONS: Respiratory retraining therapy combined with management of LPR is an effective treatment for patients with cough and PVFMD when a single-modality treatment is not sufficient. PMID- 17076098 TI - Effects of altered consciousness on the protective glottic closure reflex. AB - OBJECTIVES: The sphincteric function of the larynx, essential to lower airway protection, is most efficiently achieved through strong reflex adduction by both vocal folds. We hypothesize that central facilitation is an essential component of a bilateral brain stem-mediated adductor reflex and that its disturbance by altered consciousness or physiologic sleep could result in weakened sphincteric closure. METHODS: In 10 adult pigs the glottic closure response was evaluated under light and deep isoflurane anesthesia. The internal branch of the left superior laryngeal nerve was stimulated through bipolar platinum-iridium electrodes, and recording electrodes were positioned in the ipsilateral and contralateral thyroarytenoid muscles. The force of evoked glottic closure was measured with a pressure transducer positioned between the vocal folds. RESULTS: Consistent threshold responses (>90%) were obtained ipsilaterally from 0.5 to 2.0 minimal alveolar concentration (MAC) anesthesia. However, the contralateral reflex responses declined to 6.4% in successive trials as anesthetic levels approached 1.5 to 2.0 MAC. Furthermore, glottic closing force closely reflected these electromyographic changes, declining from 383 mm Hg at 0.5 to 1.0 MAC to 114 mm Hg at 1.5 to 2.0 MAC. CONCLUSIONS: Alteration of central facilitation by progressively deeper loss of consciousness abolishes a lower brain stem-mediated crossed adductor reflex, predisposing the subject to a weakened glottic closure response. PMID- 17076099 TI - Fine needle aspiration of the vocal fold lamina propria in an animal model. AB - OBJECTIVES: The development of a minimally invasive fine needle aspiration (FNA) technique for vocal fold (VF) biopsy would have far-reaching implications and applicability in laryngology. The objective of this study was to determine whether FNA of the VF lamina propria is feasible and whether it causes injury to the VF. METHODS: Unilateral VF FNA was performed with a 26-gauge needle on 20 rabbits. The FNA cell collection of the lamina propria was confirmed by cytology. Four weeks after the FNA, the rabbits were painlessly sacrificed and the larynges were harvested. Histologic analysis of the VF lamina propria included Masson's trichrome and elastin-van Gieson stains. The linear viscoelastic shear properties, elastic modulus, and viscous modulus of the tissue were measured. RESULTS: Compared to the contralateral normal VFs, those VFs that underwent FNA demonstrated no significant differences in levels of collagen (p = .17) and elastin (p = .94). Rheologically, the elastic shear modulus and viscous modulus did not significantly differ between the normal and FNA VFs (p = .9380 and p = .9359, respectively). CONCLUSIONS: Fine needle aspiration of the VF lamina propria is feasible without injury and offers a potentially promising, less invasive alternative to be developed for future diagnostic and therapeutic management of VF lesions. PMID- 17076100 TI - Effect of steroids on increased permeability of blood vessels of the stria vascularis after auditory ossicle vibration by a drill in otologic surgery. AB - OBJECTIVES: The vibration caused by drills used for middle ear surgery is considered one of the causes of postoperative sensorineural deafness. Seki et al reported that when drill-induced damage was created in the auditory ossicles of guinea pigs, permeability across the capillary vessels in the stria vascularis increased significantly with the duration of drill-induced vibration. The present study was undertaken to examine changes in permeability across the stria vascularis capillaries following vibration in experimental animals pretreated with steroids, with the goal of developing a method of preventing a vibration induced increase in permeability across these capillaries. METHODS: After an intravenous dose of hydrocortisone and horseradish peroxidase, the auditory ossicles of guinea pigs were vibrated with a drill for 60 seconds. RESULTS: Intravenous steroid administration before vibration reduced the leakage of horseradish peroxidase from the stria vascularis capillaries after vibration. CONCLUSIONS: The findings suggested that steroids suppress the increase in permeability across the stria vascularis capillaries that results from drill induced vibration. PMID- 17076101 TI - Relationship between time of exposure of laryngopharyngeal reflux and gene expression in laryngeal fibroblasts. AB - OBJECTIVES: Acid reflux is damaging to the laryngeal mucosa; however, the significance of the duration of reflux episodes has not been evaluated. The purpose of this study was to determine whether varying the exposure times at low pH with or without pepsin alters gene expression in laryngeal fibroblasts. METHODS: Human false vocal fold and postcricoidal cultures were exposed to pH 4 or pH 5 media with and without pepsin for 10, 30, 60, and 240 seconds. Using a real-time polymerase chain reaction, we determined the messenger RNA expression of TGFbeta-1, VEGF, FGF-2, EGR-1, ATF-3, CTGF, MMP-1, MMP-2, and decorin. RESULTS: Molecular responses were initiated at pH 5. Postcricoidal fibroblasts were more sensitive than false vocal fold fibroblasts to the presence of pepsin. Changes in transcript levels were dependent on acid exposure time, and the most significant changes were measured during the first 60 seconds after exposure. CONCLUSIONS: Time of exposure to acid and pepsin needs to be taken into consideration when determining limit of pathology in pharyngeal reflux. Genes are identified that are induced by low pH and that may be of potential importance in the pathogenesis of reflux laryngitis. PMID- 17076102 TI - Development of the human true vocal fold: depth of cell layers and quantifying cell types within the lamina propria. AB - OBJECTIVES: We sought to further describe the development of the 3-layered human vocal fold in children and to quantify macrophage and myofibroblast concentrations in each layer. METHODS: We used an optical analysis software package to examine 8 longitudinally sectioned human vocal folds that had been fixed in formalin (ages 2 days to 14 years). RESULTS: The 2-day-old vocal fold contained only a monolayer of cells. This became a bilayer by 5 months, and a trilayer began to become evident by 7 years. The percent of total depth represented by the superficial layer of the lamina propria (SLP) gradually decreased with age. The SLP made up 22% of the total lamina propria by age 7 years; this percentage approximates that in the adult vocal fold. Macrophages and myofibroblasts were predominately found in the SLP, and began to be apparent by 11 months of age. CONCLUSIONS: These results help describe the development of human voice and may have implications as to when phonosurgical therapy can be considered for children. PMID- 17076103 TI - Balloon sinuplasty: not ready for prime time. PMID- 17076105 TI - Documenting away a malpractice suit. PMID- 17076106 TI - Dress like professionals. PMID- 17076107 TI - Diagnosis du jour. PMID- 17076108 TI - Managed care administrative tasks: cutting the red tape. PMID- 17076109 TI - Sports physicals: a coding conundrum. PMID- 17076110 TI - Taming the sample closet. PMID- 17076111 TI - How to improve the quality of waived tests. PMID- 17076112 TI - Get ready for the revised CMS-1500 claim form. PMID- 17076113 TI - Comfort always. PMID- 17076114 TI - Fix Delaware's Affidavit of Merit law. PMID- 17076115 TI - The Sammons case: Lessons learned from the Affidavit of Merit. PMID- 17076116 TI - Earlier hospice referrals help patients, families, and physicians. PMID- 17076117 TI - Finishing the job. PMID- 17076118 TI - The African orphan crisis and international adoption. AB - The plight of Africa's AIDS orphans has reached crisis proportions, and the international community is beginning to mobilize at the family, community, national, and international levels. Despite these encouraging efforts, the response is inadequate, and increased attention and action are needed. The authors suggest that international adoption, although a small and temporary solution, may fit within the framework being used as a global working model. Issues surrounding adoption, such as racism, racial and cultural identity development, waiting children in the U.S. public child welfare system, and the potential legal risks, are delineated. The African perspective toward adoption is presented, although the response varies and is not collectively defined. The authors conclude that although international adoptions should be used as a last resort solution, and with tight regulations, the potential benefits to some children merit the opening of a dialogue on the topic. PMID- 17076119 TI - Social workers' attitudes about psychotropic drug treatment with youths. AB - There is considerable controversy among mental health professionals and the public about the proper role of psychotropic medications in the treatment of youths. Within social work, too, there have been sharp differences of opinion. There have been few studies, however, about the views of practicing clinical social workers on the use of psychiatric drugs in the treatment of youths. This study, a cross-sectional survey of a national sample of social workers, examines their views about medications and the role they may play in the treatment of youths.The findings suggest that social workers hold complex views that recognize both the potential benefits and harms ofpsychotropic medications, but overall they seem to support their use in a judicious manner. PMID- 17076120 TI - Financial well-being of young children with disabilities and their families. AB - Young children with disabilities are significantly more likely to live in poverty than their peers without disabilities. Exposure to poverty creates additional risk of adverse outcomes for these vulnerable children. In this article, the following key circumstances contributing to this increased risk of impoverishment are analyzed: elevated costs of raising children with disabilities, low levels of public income transfer benefits, difficulty balancing parental employment and caregiving responsibilities, unavailable or high-cost child care, and inadequate leave time to permit parents to meet their children's episodic care needs. The implications of these issues for social work practice and advocacy needs are discussed. PMID- 17076121 TI - Are we ignoring youths with disabilities in foster care? An examination of their school performance. AB - This study examined the extent to which the academic achievement of adolescents receiving both foster care and special education services differs from the performance of youths involved in only foster care, special education, or general education. Extant school data were collected on 327 students, ages 13 through 21, who attended school in a large urban school district in Oregon. The study also collected information about students' general foster care experiences, such as length of time in care and type and number of placements. Analyses revealed that foster care youths in special education typically demonstrated lower performance on academic variables in contrast to one or more of the comparison groups. These youths also appeared to experience more restrictive special education placements than youths in special education only. Although foster care or special education status alone appears to place a student at risk of academic difficulties, the negative impact of interfacing with both systems appears multiplicative. Greater attention, commitment, and time must be given to the educational needs of foster care youths with disabilities by both education and child welfare professionals. PMID- 17076122 TI - Acculturation stress and depression among Asian immigrant elders. AB - This study examines the association between acculturation stress and depressive symptoms in a regional probability sample (n = 407) of six groups of Asian immigrant elders (Chinese, Korean, Indian, Filipino,Vietnamese, and Japanese). Findings suggest that about 40 percent of the sample were depressed, indicating higher depression rates than found in other studies of older American or Asian elderly samples in the United States and Asia. Multiple regression analyses indicated that acculturation stress caused by elders' perception of a cultural gap between themselves and their adult children was associated with high depression levels. Other predictors of depression were poor perceived health, stressful life events, religiosity, proximity of children, assistance received from adult children, and longer residence in the United States. Data suggest that depression is prevalent among urban Asian immigrant elders and that there is great heterogeneity among Asian ethnic subgroups. Implications for social work practice are discussed. PMID- 17076123 TI - The animal-human bond and ethnic diversity. AB - Affectionate relationships with animal companions have health-enhancing effects on people and enrich their quality of life, and the majority of families with companion animals regard their animals as family members. Research has also suggested that these relationships are complicated and vary depending on a number of factors, yet there has been almost no exploration of ethnic diversity in relationships with companion animals. This descriptive study explores the relationships among race and ethnicity, beliefs about companion animals, and ownership practices. Findings indicate that in many instances there were no statistical differences by ethnicity. Nonetheless, describing oneself as white, American Indian, or both was associated with being more likely to have companion animals. Those identifying themselves as of Hispanic or Spanish origin were less likely to have cats and to have their cat or dog spayed or neutered and more likely to say they get a sense of personal safety from their dog or cat. The implications of these findings are discussed, and suggestions for research and practice are offered. PMID- 17076124 TI - Social work in the movies: another look. PMID- 17076125 TI - Role of the social worker in old versus new culture in nursing homes. PMID- 17076126 TI - Confucian filial piety and long-term care for aged parents. PMID- 17076127 TI - The complexities of ethical evaluation of genomics research. PMID- 17076128 TI - Practical experiences in the work of institutional ethics committees in Croatia on the example of the Ethics Committee at Clinical Hospital Center Rijeka (Croatia). PMID- 17076129 TI - Ethics committees in Croatia in the healthcare institutions: the first study about their structure and functions, and some reflections on the major issues and problems. AB - OBJECTIVES: In Croatia, ethics committees are legally required in all healthcare institutions by the Law on the Health Protection. This paper explores for the first time the structure and function of ethics committees in the healthcare institutions in Croatia. DESIGN: Cross-sectional survey of the healthcare institutions (excluding pharmacies and homecare institutions) to identify all ethics committees. SETTING: Croatia six years after the implementation of the Law on the Health Protection. MAIN MEASUREMENTS: Structure and function of ethic committees in the healthcare institutions. RESULTS: 46% of the healthcare institutions in Croatia (excluding pharmacies and homecare institutions) have an ethics committee; 89% of ethics committees have 5 members 3 of whom are from medical professions and 2 come from other fields; 49% of those committees stated that their main function is the analysis of research protocols. Only a small fraction of those ethics committees sent in standing orders, working guidelines or other documents that are connected with their work. CONCLUSIONS: Although there are legal provisions for ethics committees in the healthcare institutions in Croatia, there is an evidence of discrepancies between the practice and the "Law on the Health Protection," suggesting the need for revision of the law. There is a need for creating separate networks of HECs and IRBs in Croatia. In comparison with other countries, the development of ethics committees in Croatia has some similarities with other transitional societies in Europe. Additional research should be undertaken in the work of ethics committees in Croatia in order to understand committees' group dynamics, attitudes, and knowledge. PMID- 17076130 TI - Stark regulation: a historical and current review of the self-referral laws. PMID- 17076131 TI - The role of healthcare ethics committee networks in shaping healthcare policy and practices. PMID- 17076132 TI - Public policy's awakening to the value of home and community-based care. PMID- 17076133 TI - The Center for Health Care Law: the legal muscle of home care and hospice. AB - The Center for Health Care Law is a unique offering for a trade association. Operating as a law firm within NAHC, the Center has strengthened NAHC's advocacy efforts on all fronts. The law provides a useful structure and a rational system for behavior, provided that the law is understood. The Center brings the necessary understanding of the law to home care and hospice. PMID- 17076134 TI - Milestones in regulatory affairs. PMID- 17076135 TI - HOSPICE: An HAA/NAHC historical perspective. PMID- 17076136 TI - Celebrating the past: claiming the future. PMID- 17076137 TI - Mortality and Morbidity Liaison Committee--ISCS body mass index study. AB - OBJECTIVE: This study used the Impairment Study Capture System (ISCS) to examine the relationship between mortality and body mass index (BMI) in an insured population, particularly BMI in isolation from other risk factors. BACKGROUND: Large-scale studies of build in an insured population have traditionally been done on policies issued at standard premium rates. Insured mortality experience on elevated BMI is scarce or outdated. Increasingly competitive underwriting of build throughout the years has influenced what has been issued standard, and therefore, the relative experience of substandard to standard business. METHODS: We studied 241,966 policies submitted through the ISCS between 1989 and 2003 with actual height and weight and a code signifying abnormal build. Actual BMI were derived for these insureds. The average BMI was 35.0. Standardized mortality ratios (SMR) were computed using the 2001 Valuation Basic Table (VBT) as the expected basis. The average duration of exposure was 2.5 years. Results were stratified by underwriting factors of interest. RESULTS: Standardized mortality ratios rose quite modestly as BMI increased up until reaching severe obesity. Ratios for nonsmoker policies where elevated build was the only impairment saw SMR of 265% at BMI < 18.5, 130% at BMI 30.0-34.9, 160% at BMI 35.0-39.9 and 239% at BMI > or = 40.0. Ratios where other impairments were present tended to be higher in moderately obese ranges and lower at extremely obese ranges. No underwriting factor of significance impacted the pattern of ratios as BMI increased. CONCLUSIONS: While an average of 3 years may not be long enough to see the true manifestations of obesity, excess mortality is exhibited at low and high BMI ranges, especially when seen in isolation. The study is not a direct comparison of obese to non-obese subjects, but it is a point of reference for how obese insureds have fared vs standard issued policies. The relatively favorable experience may have more to do with the construction of the (standard) VBT table than any mitigating effect of modest obesity. PMID- 17076138 TI - Progression of mean age and mean expected mortality rate by duration of follow up in cohorts with a wide range of age. AB - BACKGROUND: [corrected] In a previous article, it was demonstrated that use of mean age to enter a life table to obtain a mean expected mortality rate of a cohort with a wide age range invariably underestimates the true mean expected mortality rate, q'. This is due to the bias introduced by the average 10% annual increase in q' in the approximate age range of 45 to 90 years (rates in the population life table, ages 0-109 years were analyzed to illustrate this). The magnitude of the error was demonstrated in various examples. All of these data were limited to the first year of FU (follow-up) duration. In this article, we analyze progression of mean age, x, and mean expected mortality rate, q', with FU duration in cohorts with all ages combined. When the age range is only 5 or 10 years, the mean age of the survivors does increase very nearly a full year with each year of FU duration. RESULTS: We utilized a 1973-1987 cohort in the SEER database for prostate cancer, all ages and all stages combined, and from this derived a comparative mortality table. We first demonstrated the difference between cumulative expected survival rate, P', as calculated in the SEER database, and the actuarial calculation of P'. The SEER method results in a 5% underestimate of P' vs the actual P' at a duration 14-15 years, and a corresponding overestimate of Q' and q'. Second, we found that the annual mean age of the survivors in the prostate cancer cohort increased from 72.4 years at entry to 80.2 years in a FU of 15 years. Mean expected q' increased from 66.7 per 1000 in the first year to 93.1 per 1000 in the 15th year. The geometric mean annual increase in mean q' was only 2.4% per year, instead of the approximate 10% seen in the life table from about age 45 to 90. Progression patterns by duration for mean age and mean q' are very different in female thyroid cancer, all ages and all stages combined, again for a 1973-1987 cohort. In thyroid cancer, females outnumber males; in both sexes, the proportion of younger patients, under age 45, is much greater than in typical cancer sites, such as prostate cancer. In female thyroid cancer, both mean age and mean q' actually decreased from the mean values at entry for 5 years or more. At entry, mean age was 43.9 years, and mean q' was 8.2 per 1000. These values decreased to 43.5 years and 6.8 per 1000, respectively, at duration 1-2 years, then leveled off and began a gradual increase. At duration 14-15 years, mean age was 53.7 years, and mean q' was 11.4 deaths per 1000 per year. CONCLUSION: Progression of mean q' is erratic and unpredictable, because annual mean age of survivors is highly dependent on the proportion of younger patients in the cohort being followed. If the proportion of patients under age 45 years is high enough, both mean age and mean q' may show an initial decrease from the values found in the year of entry, because, even though each survivor is a year older, the progression of mean age is so heavily biased by the slower progression of q' at the younger than at the older ages. However, with the SEER database, if annual expected survival rates are converted to annual expected mortality rates, the derivation of mean expected mortality rate, q' is accurate, regardless of the width of the age range in the cohort selected and being analyzed. The user of the SEER database is warned that the expected cumulative survival rate, P', is derived in the SEER survival tables on the basis of the first-year age distribution, not on the basis of the changing age distribution that is actuarially observed. PMID- 17076139 TI - Clinical implications of proteinuria. AB - Proteinuria has long been considered a marker for renal disease. New observations are suggesting that proteinuria is also a risk factor for other outcomes as well, such as cardiac events and stroke. This article will review a recommended strategy for proteinuria detection and the associations of proteinuria with the progression of renal disease and cardiovascular disease. It will also review the significance of the development of proteinuria in the elderly population and discuss other common causes of proteinuria apart from those secondary to diabetes and hypertension. PMID- 17076140 TI - Severity criteria for critical illness insurance definitions. AB - BACKGROUND: Definitions for critical illness insurance must be updated to reflect increasing incidence of covered events and improvements in screening, diagnosis and treatment. METHODS: Examples are provided for ways to future-proof definitions via use of severity criteria. RESULTS: The desired effect could be achieved by adding severity criteria as follows: myocardial infarction--ejection fraction, troponin level, or functional class; stroke--Modified Rankin Scale; and cancer--scaled benefits in proportion to probability of survival. CONCLUSION: Severity criteria can limit the increase in critical illness claims that is expected based on improvements in screening and diagnostic tests, thereby keeping the price affordable while retaining the features that have made this product so successful. PMID- 17076141 TI - The Gang of Four: insurance medicine--art, science or pseudo-science? PMID- 17076142 TI - Testicular microlithiasis. PMID- 17076143 TI - Increased QRS voltage in the limb leads. AB - Increased QRS voltage accompanied by repolarization abnormalities in a life insurance applicant's electrocardiogram should alert the medical director to the possibility of left ventricular hypertrophy. If confirmed, left ventricular hypertrophy or increased left ventricular mass is a strong independent risk factor for future cardiac events and all cause mortality. The use of the electrocardiogram to diagnose left ventricular hypertrophy can be helpful, but there are limitations that need to be considered. This ECG case study illustrates some of the pitfalls in the electrocardiographic diagnosis of left ventricular hypertrophy. PMID- 17076144 TI - Erroneous diagnosis of myocardial infarction--revisited. 1982. PMID- 17076145 TI - Ebstein's anomaly and pre-excitation of the WPW type--revisited. 1982. PMID- 17076146 TI - Pick's disease. PMID- 17076147 TI - Hereditary hemorrhagic telangiectasia. AB - Hereditary hemorrhagic telangiectasia is an inherited condition characterized by abnormal blood vessels found in the mucosa, skin and internal organs, which are delicate and prone to bleeding. PMID- 17076148 TI - Hypothenar hammer syndrome. AB - Thrombosis of the ulnar artery in the hand is the most common type of upper extremity vascular occlusion. Often caused by constant pounding on the ulnar side of the palm, it is known as hypothenar hammer syndrome. PMID- 17076149 TI - The legacy of Dr. Rene Laennec. PMID- 17076150 TI - [A failure disguised as victory... and which soon will target practitioners]. PMID- 17076151 TI - [Immunotherapy for head and neck squamous cell carcinoma]. AB - In the past decades, prognosis of head and neck squamous cell carcinoma (HNSCC) has not improved despite substantial progress in treatment options. Since antitumoral immunity was described, immunotherapy has shown promising results as an adjunctive treatment in various cancer types. Tumor-associated antigens (TAAs) have been identified and shown to stimulate selective T-cell-mediated antitumoral immune response. This article briefly reviews the work done in the field of immunotherapy of HNSCC in the past few years. It gives confidence that immunotherapy may play an important role in the treatment of head and neck squamous cell carcinoma. Among various TAAs, the family of cancer testis antigens (CTAs) may be promising candidates for specific immune therapy in HNSCC. Ongoing studies will confirm whether CTAs may generate an immune response in clinical vaccine trials. PMID- 17076152 TI - [Diagnosis and management of facial paralysis]. AB - Facial palsy is an unusual pathology that requires standard investigations and management. A clinical overview of the current attitudes is suggested to the general practitioners in order to help them in initiating the adequate investigations and treatment before referring the patient to a specialist. PMID- 17076153 TI - [Professional occupation after cochlear implantation]. AB - This study verifies whether cochlear implants helps deaf adults to maintain or develop their professional occupations. Sixty-seven patients received a questionnaire concerning their professional activities before and after implantation. At the time of implantation 34 were professionally active. After the implantation 29 remained active, 4 of them reporting positive developments in their careers. Five patients became inactive. The previously inactive patients remained inactive. There was no difference in auditory performances between professionally active or inactive patients. Cochlear implants enable most implanted adults to maintain and even progress in their professions. However, deafness still represents an obstacle to social integration as inactive patients who searched for a job were rejected after the job interviews. PMID- 17076154 TI - [Cochlear implant or regular hearing aid?]. AB - A significant number of deaf patients that have received cochlear implants now achieve higher word recognition scores then those with conventional auditory prostheses. This situation makes the choice of which type of auditory rehabilitation to propose a complex matter in patients with remaining auditory function. Our paper aims at providing some arguments to these new questions by presenting the clinical experience and practice of the Centre romand d'implants cochleaires. We also address related legal issues. Clinical tools, such as testing the comprehension of lists of logatoms have proved very useful for the evaluation of these particular patients. The evaluation of cochlear implant candidates remains a highly individualized process, necessitating a case by case approach by an experienced multidisciplinary cochlear implant team. PMID- 17076155 TI - [Difficulties encountered by old patients wearing a hearing aid]. AB - Presbycusis is a common cause of deafness in the elderly. The histopathology shows degenerative changes of the hair cells in the organ of Corti, and of the auditory nerve fibers. This implies a loss of sensibility for low intensity stimuli, although the perception of loud sounds remains normal. The dynamic range of the cochlea is therefore reduced. In addition, the discrimination is also altered. Hearing aids do not restore a normal hearing. Manufacturers have reduced their size, making them difficult to handle by old patients. This paper reports some of the difficulties encountered by old patients suffering from presbycusis, their family members, doctors, and nurses are not always aware of. PMID- 17076156 TI - [Gender, ages, psychotropic and anti-pain drugs uses in the Third Swiss Health Survey]. AB - From the third Swiss Health Survey, gender prevalence of psychotropic and anti pain drugs uses are described. Age differences are added to gender differences. In particular for aging populations, psychotropic drugs are prescribed by compassion. Results suggest medical doctors as prescriptors attribute a large impact to psychological suffering coming from physical illness and incapacities linked to age. Analysis of drug uses allows fruitful interpretation of the patient-doctor relationship by gender and age. PMID- 17076158 TI - [What talking means. Remarks on bullshit]. PMID- 17076157 TI - [The striking pressure of Mr. B]. PMID- 17076159 TI - [Evidence of the efficacy of eculizumab in paroxysmal nocturnal hemoglobinuria]. PMID- 17076160 TI - [203 years later, thanks to Jan Teerlink]. PMID- 17076161 TI - [Human stem cells: the appeal of the Vatican]. PMID- 17076162 TI - [High risk swimming-pool]. PMID- 17076163 TI - Patient's page. Bottled water and your teeth. PMID- 17076164 TI - Dental care for the aging population. PMID- 17076165 TI - Multicentric radiolucencies between teeth. PMID- 17076166 TI - The state of state RHIOs. A one-size-fits-all approach won't work for regional health information organizations. PMID- 17076167 TI - Migration nation. Clinical workflow takes priority as organizations improve processes with technology. PMID- 17076168 TI - Start your engines. Providers work on integration to facilitate messaging between applications. PMID- 17076170 TI - The secrets of successful CIOs. Top CIOs talk about what it takes to lead so others want to follow. PMID- 17076169 TI - ICD-10 on the way? Despite fierce opposition from some powerful industry groups, ICD-10 codes could be coming fast. PMID- 17076171 TI - Bedside manners. Mobile computing goes bedside to increase safety at the point of care. PMID- 17076172 TI - Programmed for innovation. The Innovation Program at Partners HealthCare ignites creativity in the young and talented. PMID- 17076173 TI - Why are we doing this? How leading with technology can cause unintended negative consequences. PMID- 17076174 TI - Let's get going. Health systems should start focusing on how to best share diagnostic images. PMID- 17076175 TI - Shifting the burden. Web-enabled shift scheduling technology may be an easier way to handle a very complex task. PMID- 17076176 TI - It takes a community. Health information technology systems are good for community health centers. PMID- 17076177 TI - [Study on the relationship between childhood obesity and various determinants, including socioeconomic factors, in an urban area]. AB - OBJECTIVES: This study was conducted to investigate the prevalence of childhood obesity, the association between the undesirable lifestyles and socioeconomic factors, the association between childhood obesity and various risk factors, including socioeconomic factors, and the agreement between the body mass index (BMI) classification and the body fat percentage. METHODS: The study subjects were all the 5th grade students from all the elementary schools in Gunpo City, Kyunggi Province, South Korea (4043 children at 22 schools). The subjects were measured for their height, weight and percent body fat etc. and they were also surveyed by questionnaire from March 18th to April 25th, 2005. To determine whether the children were within normal limits or not, standardized BMIs for each age group were used. The data was analyzed by logistic regression analysis using SAS 9.0 version. RESULTS: The prevalence of childhood obesity prevalence was 25.1%. Boys had a higher prevalence of obesity (27.5%) than did the girls (22.5%). Children had tendencies of having undesirable lifestyles and getting obese if they had a lower socioeconomic status. The risk factors for childhood obesity were low paternal education (OR: 1.17, 95% CI: 0.97-1.42) and non parental caregivers (OR: 1.34, 95% CI: 0.98-1.82). Other risk factors for childhood obesity were a high birth-weight, longer TV/computer-using time, a lower fruit-eating frequency, short sleeping hours and parental obesity. The agreement rate between the BMI classification and the body fat percentage was 93.1%. CONCLUSIONS: This study showed the children had a higher prevalence of obesity: further, not only individual lifestyles, but also socioeconomic factors could influence childhood obesity. Childhood obesity was especially more problematic for children with a low socioeconomic status. PMID- 17076178 TI - [The determinants of adolescent smoking by gender and type of school in Korea]. AB - OBJECTIVES: This study assessed the influences of various factors that are related to youth smoking such as gender, age and type of school, and we wanted to provide supporting data for tailored and effective policy initiatives to reduce adolescent smoking. METHODS: A self-report survey was conducted on 14,910 teen age students who were selected based on the nationwide distribution of students in large and small cities and counties, the gender ratio of the students and the ratio of students attending various type of school at 38 middle and high schools in six representative areas of each province. The survey was handled and managed by a health education teacher at each school. Binary and multinomial logistic regression was used in the analyses. RESULTS: Smoking by adolescents was associated with gender, age and even height. Male high school students tended to smoke more than female high school students, but this differences was not significant for middle school students. The older the adolescents were, the more likely that they smoked, except for the female high school students. Height was meaningful for all adolescents, except for the boys at the vocational high schools. Monthly allowance was significant for all adolescents. School factors such as type of school and the students' school performance were also crucial factors. Attending a vocational high school was strongly related to smoking, especially for girls. Students' school performance and the perceived level of stress were strongly associated with smoking, especially for boys. Home factors such as the relationship with parents and conversation time with family members were closely related to smoking behavior. Knowledge about the health hazard of smoking was also found to be strongly related to adolescent smoking. CONCLUSIONS: In conclusion, demographic factors, school factors, home surroundings and the perception on the harmfulness of smoking are strongly related to adolescent smoking behavior, but these differ from gender and type of school. PMID- 17076179 TI - [Prevalence and associated factors of osteoporosis among postmenopausal women in Chuncheon: Hallym Aging Study (HAS)]. AB - OBJECTIVES: A community-based, cross-sectional survey was conducted to determine theprevalence of osteoporosis and to evaluate the effects of body composition, health behaviors and reproductive history on bone density in postmenopausal women. METHODS: The study subjects were 362 postmenopausal women, aged 45 years old or over, who were invited to the hospital. Information on their socio demographic characteristics and the potential risk factors such as their past medical history, smoking, alcohol intake, exercise, diet and menstrual/reproductive histories were collected by trained interviewers. Weight, height, the body mass index (kg/m2), and body composition variables were measured. Bone mineral density of the lumbar spine was measured by dual energy X ray absorptiometry (DXA). RESULTS: The prevalence of osteoporosis was 30.6% in the 45-64 years old women, 52.5% in the elderly women aged 65-74, and 68.7% in the women aged 75 years or over. After adjustment for the effect of potential covariates, those women in the highest 25% (4th quartile) of the lean body mass are less likely to have osteoporosis (aOR=0.31, 95% CI=0.12-0.76), compared with the lowest quartile group. More parity also had significantly detrimental effects on osteoporosis. CONCLUSIONS: These findings suggest that the prevalence of osteoporosis in postmenopausal women increased with age from 46.3% of those aged 45-64 to 68.7% for those aged 75 and over. Lean body mass and parity appeared significant contributor to bone mineral density in postmenopausal women in this population. PMID- 17076180 TI - [Cost-of-illness study of asthma in Korea: estimated from the Korea National Health insurance claims database]. AB - OBJECTIVES: We estimated the asthma-related health care utilization and costs in Korea from the insurer' s and societal perspective. METHODS: We extracted the insurance claims records from the Korea National Health Insurance claims database for determining the health care services provided to patients with asthma in 2003. Patients were defined as having asthma if they had > or =2 medical claims with diagnosis of asthma and they had been prescribed anti-asthma medicines. Annual claims records were aggeregated for each patient to produce patient specific information on the total utilization and costs. The total asthma-related cost was the sum of the direct healthcare costs, the transportation costs for visits to healthcare providers and the patient's or caregivers' costs for the time spent on hospital or outpatient visits. RESULTS: A total of 699,603 people were identified as asthma patients, yielding an asthma prevalence of 1.47%. Each asthma patient had 7.56 outpatient visits, 0.01 ED visits and 0.02 admissions per year to treat asthma. The per-capita insurance-covered costs increased with age, from 128,276 Won for children aged 1 to 14 years to 270,729 Won for those aged 75 or older. The total cost in the nation varied from 121,865 million to 174,949 million Won depending on the perspectives. From a societal perspective, direct healthcare costs accounted for 84.9%, transportation costs for 15.1% and time costs for 9.2% of the total costs. CONCLUSIONS: Hospitalizations and ED visits represented only a small portion of the asthma-related costs. Most of the societal burden was attributed to direct medical expenditures, with outpatient visits and medications emerging as the single largest cost components. PMID- 17076181 TI - [One year follow-up study of symptomatic cases of ulnar neuropathy at the elbow in a rural population]. AB - OBJECTIVES: This study examined the natural history of symptomatic patients who did or did not display abnormal results on nerve conduction studies (NCS). METHODS: Forty hundred fifty adults were selected among a total of 578 residents who participated in the health examination in a rural Korean district. A symptom questionnaire and NCS were used to diagnose ulnar neuropathy at the elbow (UNE). There were 6.4% of the subjects with UNE, 5.1% of the subjects showed symptoms without a NCS, and 84.2% of the subjects who were asymptomatic. One year later, 20 symptomatic limbs with an abnormality on the ulnar NCS and 22 symptomatic limbs without any abnormality in the ulnar NCS were enrolled in a follow-up study. The natural history of UNE was evaluated by examining the changes in the clinical and electrodiagnostic examinations. RESULTS: The 1-year follow-up of the enrolled limbs found that for the symptomatic limbs with an abnormality on the NCS, the degree of severe of the clinical grade changed from 20% to 10%. In contrast, for the symptomatic limbs that were without any abnormality in the NCS, the change of the severe degree of the clinical grade was from 0% to 18.2%. Also, for the electrodiagnostic change, only symptomatic limbs without NCS abnormalities showed significant changes in motor latency, amplitude and conduction velocity at the 1-year follow-up. CONCLUSIONS: The 1-year follow-up study revealed symptomatic limbs that were without any abnormality on the ulnar NCS were more likely to progress than the symptomatic limbs with an abnormality on the ulnar NCS. PMID- 17076182 TI - [Differences in medical care utilization rates of the disabled and the non disabled with ambulatory care sensitive conditions]. AB - OBJECTIVES: The purpose of this study was to determine whether the disabled have worse access to primary care than the non-disabled. METHODS: We used the National Disability Registry data and the National Health Insurance data for the calendar year 2003, and we analyzed 807,380 disabled persons who had been registered until December 2001 and we also analyzed 1,614,760 non-disabled persons for nine ambulatory care sensitive conditions (ACSCs). The rates of physician visits and hospitalizations for the patients with ACSCs were compared between the disabled and the non-disabled. Multiple logistic regression analysis was used to evaluate the association between medical care utilization and disability and to assess the association between hospitalization and the number of physician visits while controlling for potential confounders. RESULTS: The numbers of physician visits per 100 patients were 0.78-0.97 times lower for the disabled than that for the non-disabled with five of nine ACSCs. The numbers of hospitalizations per 100 patients were 1.16-1.77 times higher for the disabled than that for the non disabled with all the ACSCs. While the ORs of a physician visit for the disabled were significantly lower than that for the non-disabled with all the ACSCs (OR: 0.44-0.70), and the ORs of hospitalization for the disabled were significantly higher (OR: 1.16-1.89). The lower physician visit group (number of physician visits < or =1) was more likely to be hospitalized than the higher physician visit group (number of physician visits > or =2) (OR: 1.69-19.77). The effect of the physician visit rate on hospitalization was larger than the effect of disability on hospitalization. CONCLUSIONS: The results suggest that the disabled were more likely to be hospitalized for ACSCs due to their lower access to primary care. PMID- 17076183 TI - [Relationships between mental health and psychosocial factors with single-child high school students in an urban city of Korea]. AB - OBJECTIVES: This study was performed to determine the mental health of high school students, and specifically that of children with no siblings in urban areas, and we aimed at revealing the various potential influences of different psycho-social factors. METHODS: The participants were, 514 high school students who were the 1st- to 3rd-graders in Daejon City; they were, given self administered questionnaires that required no signature during the period of March through June 2005. The analyzed items included the general character of the subjects, the symptoms of stress and depression for mental health, self-esteem as a psychological component, anxiety, dependent behavioral traits and, social support of family members and friends. RESULTS: The study results suggested that the group of urban high school children with no siblings had a higher tendency for stress and depression than did the urban high school children with siblings. The mental health and psychosocial factors were found to be influenced by friends, a sense of satisfaction at school and home life, and emotional support as well. CONCLUSIONS: In conclusion, emotional support by the family members can improve mental health by reducing anxiety, stress and depression. PMID- 17076184 TI - Dipstick urine protein, as a predictor of cardiovascular mortality in Korean men: Korea Medical Insurance Corporation study. AB - OBJECTIVES: This study was to investigate if the dipstick proteinuria can predict cardiovascular mortality in a population of Korean men. METHODS: We measured urine protein and other cardiovascular risk factors in 100,059 Korean men, aged between 35-59 years in 1990 and 1992. Levels of proteinuria measured by dipstick method were trace or less, 1+, 2+, and 3+ or greater. The primary outcomes were deaths from all causes, cardiovascular disease, cancer, and others in a 12 year follow-up from 1993 to 2004. RESULTS: The multivariate-adjusted relative risks (95% CI) for cardiovascular death according to the level of proteinuria (1+, 2+, 3+ and more) in 1990 examination were 2.18 (1.36-3.48), 2.55 (1.37-4.78), and 4.57 (2.16-9.66) respectively. The corresponding relative risks according to the level of proteinuria in 1992 examination were 2.49 (1.71-3.64), 2.64 (1.53-4.58), and 2.78 (1.15-6.73). The relative risks for cardiovascular death of men with proteinuria (1+ or greater) once and twice among the examinations were 2.18 (1.63 2.92) and 3.75 (2.27-6.18), compared with men without proteinuria in 1990 and 1992 examinations. CONCLUSIONS: Our results showed that dipstick proteinuria is associated with cardiovascular mortality in Korean men. Dipstick proteinuria could be a predictor for cardiovascular mortality. PMID- 17076185 TI - [Estimation of a nationwide statistics of hernia operation applying data mining technique to the National Health Insurance Database]. AB - OBJECTIVES: The aim of this study is to develop a methodology for estimating a nationwide statistic for hernia operations with using the claim database of the Korea Health Insurance Cooperation (KHIC). METHODS: According to the insurance claim procedures, the claim database was divided into the electronic data interchange database (EDI_DB) and the sheet database (Paper_DB). Although the EDI_DB has operation and management codes showing the facts and kinds of operations, the Paper_DB doesn't. Using the hernia matched management code in the EDI_DB, the cases of hernia surgery were extracted. For drawing the potential cases from the Paper_DB, which doesn't have the code, the predictive model was developed using the data mining technique called SEMMA. The claim sheets of the cases that showed a predictive probability of an operation over the threshold, as was decided by the ROC curve, were identified in order to get the positive predictive value as an index of usefulness for the predictive model. RESULTS: Of the claim databases in 2004, 14,386 cases had hernia related management codes with using the EDI system. For fitting the models with applying the data mining technique, logistic regression was chosen rather than the neural network method or the decision tree method. From the Paper_DB, 1,019 cases were extracted as potential cases. Direct review of the sheets of the extracted cases showed that the positive predictive value was 95.3%. CONCLUSIONS: The results suggested that applying the data mining technique to the claim database in the KHIC for estimating the nationwide surgical statistics would be useful from the aspect of execution and cost-effectiveness. PMID- 17076186 TI - [Developing the predictive model for the group at high risk for colon cancer]. AB - OBJECTIVES: We developed the predictive model for the incidence of colon cancer by utilizing the health screening data of the National Health Insurance in Korea. We also explored the characteristics of the high risk group for colon cancer. METHODS: The predictive model was used to determine those people who have a high risk for colon cancer within 2 years of their NHI health screening, and we excluded the people who had already been treated for cancer or who were cancer patient. The study population is the insured of the NHI, aged 40 or over and they had undergone health screening from the year 2000 to 2004, according to NHI health screening formula. We performed logistic regression analysis and used SAS Enterprise Miner 4.1. RESULTS: This study shows that there exists a higher rate of colon cancer in males than females. Also, for the population in their 60s, the incidence rate of colon cancer is much higher by 5.36 times than that for those people in their 40s. Amongst the behavioral factors, heavy drinking is the most important determinant of the colon cancer incidence (7.39 times in males and 21.51 times in females). CONCLUSIONS: Our study confirms that the major influencing factors for the incidence of colon cancer are drinking, lack of exercise, a medical history of colon polypus and a family history of colon cancer. As a result, we can choose the group that is at a high risk for colon cancer and provide customized medical information and selective management services according to their characteristics. PMID- 17076187 TI - [Analysis of leptin pulses in serum in women with polycystic ovary syndrome]. AB - OBJECTIVES: Authors estimate the influence of body mass on the leptin serum concentrations changes in course of 24 hours in twelve women with polycystic ovary syndrome (PCOS). MATERIAL AND METHODS: The course of leptin pulses was analysed in two volunteers' six-persons groups, both with PCOS. first group with BMI < 22 kg/m2 and the second group with BMI >26 kg/m2. The blood samples (0,5 ml) were taken from cubital vein in 15-20 minutes intervals through 48 hours. In the first day of clinical experiment three meals were accepted but in the second day the women were starved and they could drink only mineral water. RESULTS: The significant difference in average leptin concentration in any woman's group between each experimental day was observed. The value of leptin pulses amplitude compared between both women's group on each experimental daytime was significant too. The results we got relating number of leptin pulses and their duration time compared between both women's group and into groups between night and day were not statistically significant. CONCLUSION: For leptin pulses detection in blood the cluster method was applied, which confirmed the pulses appearance in leptin secretion in both woman's group. PMID- 17076188 TI - [Laparoscopic assessment following failure to achieve pregnancy after intrauterine inseminations in patients with normal hysterosalpingograms]. AB - OBJECTIVES: To assess the diagnostic benefit of laparoscopy in infertile women thought to be at low risk for altered pelvic anatomy. DESIGN: Retrospective chart review. MATERIAL AND METHODS: PATIENTS: 127 infertile patients who underwent laparoscopic evaluation of the pelvis failing to conceive after intrauterine inseminations (IUI) with normal hysterosalpingography (HSG). INTERVENTION: Diagnostic and/or therapeutic laparoscopy. MAIN OUTCOME MEASURES: Presence of pelvic pathology and predictors of pelvic disease. RESULTS: Although the hysterosalpingograms were read as normal in all women, endometriosis stage I-II was found in 64 (50,4%) patients, stage III and stage IV in 4 (3,1%). Adhaesions were diagnosed laparoscopically in 22 (17%) patients and distal tubal disease in 26 (20%). All of this abnormalities were directly treated by laparoscopic intervention. The time between HSG and laparoscopy was positively correlated with appearance of distal tubal disease and pelvic adhaesions. CONCLUSION: Laparoscopic findings could lead to a change of treatment decisions in high number of infertile patients with normal hysterosalpingography. PMID- 17076189 TI - [DNA ploidy and results of first line chemotherapy in ovarian cancer patients]. AB - AIM OF THE STUDY: The prognostic significance of DNA ploidy in ovarian cancer patients determined by flow cytometric analysis, in correlation with effectiveness of first line chemotherapy. MATERIAL AND METHODS: DNA ploidy by FC was investigated in group of 102 ovarian cancer patients from fresh frozen samples (4 patients was excluded from the study). RESULTS: Positive answer for first line treatment we notified in 64(62,75%) cases, lack of answer 34 (37,25%) patients. Aneuploidy was more frequent in negative group 31(91,18%), diploid tumours occurred in 3(8.82%) cases. In positive group aneuploid tumours occurred in 29(45,31%) and diploid in 35(54,69%) patients (p<0,001). Median survival in positive group--45 months, in negative group 12 months (p<0,0001). In positive group median survival in patients with aneuploid tumours--31 months, in patients with diploid tumours median survival was not reached (p=0,0102). In negative group DNA ploidy has no impact on survival (p=0,1027) CONCLUSIONS: 1. DNA ploidy determined by flow cytometry is prognostic factor in ovarian cancer patients who answered positively for first line treatment. 2. Aneuploid tumours appear much often then diploid in group of patient who did not answer for first line chemotherapy 3. Patients with diploid tumours have better prognosis. 4. Lack of positive answer for first line treatment is bad prognostic factor. PMID- 17076190 TI - [Clinical value estimation of the Doppler indicies: PI, RI and PSV, TAMXV and MEDV in early differentiation of the ovarian tumors malignancy]. AB - THE OBJECTIVES: The aim of this study was to evaluate the clinical value of blood flow indicies for differentiating between benign and malignant ovarian tumours. MATERIALS AND METHODS: Realisation of the aim was based on the analysis of examinations' results of 686 ovarian tumors. For each recorded blood flow wave the followind parameters of examined tumor vessels were analysed: pulsatility index (PI), resistance index (RI), peak systolic velocity (PSV), time-averaged maximum velocity (TAMXV) and maximum and diastolic velocity (MEDV). Apart from conditional probabilities we also presented assessing areas under the ROC curves, which introducted information over practical usefulness of each test. RESULTS: Vascular perfusion was observed in 100% of malignant tumors and 92% of non malignant tumors. In group of patients with malignant tumors PI and RI were significantly lower; and PSV, TAMXV and MEDV were higher than in group with non malignant tumors. (p<0.05) CONCLUSIONS: Transvaginal ultrasonography with the Doppler technique is valuable diagnostic method in preoperative differentiation of tumor malignancies. The best predictive value in analysed group was observed for PI (AUROC=o.87) and MEDV (AUROC=0.86). Similar correlation was observed for premenopausal patients. In group of postmenopausal patients the best predictive value was for RI (AUROC=0.90) and MEDV (AUROC=0.88). PMID- 17076191 TI - [Concentration of annexin V in gestational tissues: placenta, fetal membranes, myometrium and amniotic fluid]. AB - THE BACKGROUND: It is believed that annexin V, an anticoagulant protein abundant in trophoblast, prevents circulating blood in the intervillous space from hypercoagulability and fibrin deposition. Distribution and the role of annexin V localized in other gestational tissues like myometrium and fetal membranes, as well as in amniotic fluid is unknown. The aim of this study was to determine the concentration of annexin V in the tissues under consideration. STUDY DESIGN: The study group consisted of 40 healthy women at the 1st stage of labour, 16 of whom delivered by cesarean section. The strips of myometrium, placenta and fetal membranes, as well as the samples of amniotic fluid and the venous blood, were collected during surgery. Homogenates were prepared from the tissues. Annexin V was measured by immunoenzymatic method (ELISA). RESULTS: Concentration of annexin V in the placenta was 122.65 +/- 33,14 ng/mg protein, in fetal membranes 136.31 +/- 49.30 ng/mg protein and in myometrium 65.40 +/- 30.72 ng/mg protein. There was a statistical difference between the concentrations in placenta and fetal membranes vs. myometrium (p<0.05). In amniotic fluid annexin V was found to be in low concentration (4.46 +/- 2.59 ng/ml, i.e., 1.60 +/- 1.21 ng/mg protein) while in blood plasma it was extremely low, over 6000 times lower than in placenta. CONCLUSION: Annexin V is present not only in placenta, but also in fetal membranes and myometrium. Its concentration in fetal membranes is similar to that in placenta, while in myometrium it is 50% lower. Annexin V found in newly discovered sites probably plays a similar role like placental annexin V. PMID- 17076192 TI - [Obstetric outcome and behavior of women after transabdominal isthmocervical cerclage]. AB - OBJECTIVES: The aim of this study was to evaluate the course of the life in women with preconception transabdominal cervicoisthmic cerclage. MATERIALS AND METHODS: The studied material consisted of 9 women with cervical incompetence treated with transabdominal cervicoisthmic cerclage in period 2001-2005 in II Department of Gynaecology and Obstetrics of Medical University in Lodz. We paid special attention to mental condition. RESULTS: In analysed group 7 women got pregnant. Actually 6 of them have children and 1 is in 32 weeks pregnant. CONCLUSIONS: 1. Women after repetitive abortion or premature delivery because of serious defect of cervical isthmus are likely to suffer from depression. 2. Management with transabdominal cervicoisthmic cerclage makes carrying pregnancy up to term possible. 3. Prophylactic transabdominal cervicoisthmic cerclage left after childbirth seriously improves feeling of this women. 4. Women with transabdominal cervicoisthmic cerclage left after birth don't complain about dysurical symptoms. PMID- 17076193 TI - [Is low pre-pregnancy body mass index a risk factor for preterm birth and low neonatal birth weight?]. AB - OBJECTIVES: A pregnant woman's weight is an extremely important factor in the course of pregnancy and delivery. Not only obesity but also being underweight may lead to complications in pregnancy such as: preterm delivery and low neonatal birth weight. DESIGN: The aim of this study was to analyze the relationship between a low BMI and outcome of pregnancy, birth weight and general well being of the neonates. MATERIAL AND METHODS: 415 patients who were hospitalized in the Department of Obstetrics and Reproduction Wroclaw Medical University between 1996 2005 was done. The patients were divided into 3 groups I--Underweight (BMI <19,8), II--Appropriate weight (BMI 19,8-26,0) and III--Overweight (BMI>26,0). RESULTS: The frequency of preterm deliveries as well as low neonatal birth weight <2500g, in underweight mothers was higher than in other groups. CONCLUSIONS: Low pre-pregnancy BMI is an important factor risk factor in preterm deliveries. There was no correlation between BMI and the general well being of the neonates. PMID- 17076194 TI - [Insulin resistance in pregnant type 1 diabetic female. Case report]. AB - Insulin resistance may coexist with diabetes type 1 and make treatment of diabetes difficult. Case of 24-year-old type 1 diabetic female with insulin resistance features prior to pregnancy is reported. Exacerbating of insulin resistance during pregnancy was manifested by difficulties to overcome excessive weight gain and necessity to initiate treatment with high doses of insulin. The treatment was based on diet with progressive caloric restriction to 800 kcal/day in 35 week of pregnancy. That diet was continued till the delivery in 37 week. The fast acting analog insulin (Humalog) and long acting insulin (Humulin U) were used in treatment of diabetes. Treatment with low calorie diet did not cause negative effects on diabetic female metabolism and on the neonate state. PMID- 17076195 TI - [Spontaneus ductal closure in a fetus postnatally diagnosed as Adams-Olivier syndrome]. AB - In utero isolated ductal closure is uncommon and can lead to congestive heart failure, fetal hydrops and death if not recognized. A case report of premature spontaneus ductal closure in the third trimester of pregnancy in a fetus postnatally diagnosed as Adams-Olivier Syndrome is presented. On ultrasound examination an intrauterine growth restriction, defects of bones of hands and feet as well as ventriculomegaly were found. No nonsteroid drug treatment during pregnancy was applied. Fetal echocardiography was performed following an abnormal four-chamber view. Premature ductal closure was diagnosed. Fetal echocardiogram showed absent flow in the ductus arteriosus, dilated right ventricle with decreased function, and moderate tricuspid and pulmonary valve insufficiency with no signs of fetal hydrops. An elective cesarean section was performed. All abnormalities observed on former echocardiogram exam withdrew within 3 months of infant's life. The infant stays in the tertiary care centre due to the extracardiac malformations. PMID- 17076196 TI - [Anorexia nervosa--new view on neuroendocrine and genetic determinations]. AB - Anorexia nervosa is an eating disorder, characterized by low body weight, distorted body image, amenorrhea and an intense fear of gaining weight. The occurrence of anorexia nervosa has increased over the past 10 years among adolescents and young women and it is estimated to occur in 0,5-1% of population. The Anorexia nervosa is not only a psychiatric illness may have many serious gynecological and medical ramifications. Preventive measures to reduce the incidence of anorexia are not known at this time. However, early detection, intervention and cooperation between many specialists can reduce the severity of symptoms and health consequences. Gynecologists assume a broader role in preventative medicine and health maintenance, that is why their awareness of anorexia nervosa is needed. Anorectic patients have metabolic and endocrine complications. Most of them are caused by the dysfunction of hypothalamus, which produces many nueropeptides and neurotransmitters. Anorexia nervosa is characterized by numerous aberrations in neuropeptides and neurotransmitters, such as gonadotropin-releasing hormone, corticotropin-releasing hormone, neuropeptyd Y, leptin, beta-endorfins and serotonine, dopamine. The relationship of anorexia nervosa with genetic factor is being enhanced lower the last few years. However, the studies on the role of polymorphism in some genes brought conflicting results. PMID- 17076197 TI - [The role of peroxisome proliferator-activated receptors (PPAR) in carcinogenesis]. AB - Peroxisome proliferators-activated receptors are members of the nuclear hormone receptor superfamily of ligand-activated transcription factors. The PPAR subfamily consists of three members: PPAR-alpha, PPAR-sigma (NUC-1 or beta) and PPAR-gamma. PPARs regulate gene expression by binding, as heterodimers with retinoid X receptors (RXR), to specific response elements (PPREs) in the promoter regions of target genes. The prostaglandin 12 especially, all arachidonic acid metabolites and polyunsaturated fatty acids are naturally occuring PPAR ligands. Synthetic PPAR ligands are thiazolidinediones (TZDs--rosiglitazone, pioglitazone, troglitazone). Activation of nuclear hormone receptors has been identified as an approach to induce differentiation and inhibit proliferation of cancer lines. The anti-proliferative, pro-differentiation effects of PPAR activators (TZDs) suggest that these compounds might be useful in slowing the proliferation of un differentiated tumour cells. TZDs inhibit proliferation of human breast, prostate and colon cancers, both in vitro and in tumours derived from these cells implanted into rodents. Furthermore, recent studies show that PPAR-gamma ligands are potent inhibitors of angiogenesis, a process essential for solid-tumour growth and metastasis. In conclusion, the evidence to date suggests that activation of PPAR should suppress tumour growth and development. This represents an exciting novel therapeutic application of TZDs. In present paper, structural features of PPARs, their gene transcription mechanisms and recent developments in the discovery of their biological functions are reviewed. PMID- 17076198 TI - [Polish Gynecological Society's recommendations regarding ovarian cancer]. PMID- 17076199 TI - [Laparoscopy or laparotomy in the treatment of ovarian teratomas?]. AB - BACKGROUND: Mature cystic teratomas (dermoid cyst), are the most common germ cell tumours of the ovary in women of reproductive age. They are usually asymptomatic until they reach considerable size. Frequently, ultrasonography is diagnostic. MATERIALS AND METHODS: 81 patients who had surgery in 3rd Departament of Gynecology and Obstetrics, University of Medicine, Lodz, from 2000 to 2005 for benign cystic teratomas were analyzed. RESULTS: 86 teratomas were removed from 81 patients. 31 patients had surgery via laparoscopy and 50 via laparotomy. The mean age was found to be 34,5 years. 39 patients were asymptomatic. The tumours diameter ranged from 1 to 19 cm. In 46 cases there were cystectomies performed sparing fragments of healthy ovarian tissue, in 24 cases ovariectomies and in 11 panhysterectomies were made. All removed lesions histologically appeared benign cystis ovarian teratomas without any signs of malignancy. CONCLUSIONS: The teratomas are composed of mature tissues derived from the three germ cell layers. In our study histologic examination revealed 16 cases of neurogenic tissue. One of the possibility of ovarian teratoma's treatment is operative laparoscopy. When the laparoscopy is not possible or inadvisable, the laparotomy is still irreplaceable. PMID- 17076200 TI - [Discrepancies between clinical and surgical--pathologic staging of cervical carcinoma]. AB - OBJECTIVES: Cervical cancer is the only gynecological tumor staged clinically. The aim of this study was comparison of clinical staging to surgical specimen evaluation in cervical cancer patients. DESIGN: Clinical staging of cervical cancer was compared to histopathological outcome in prospective study. MATERIALS AND METHODS: One hundred patients with cervical cancer treated with radical hysterectomy combined with pelvic lymphadenectomy were included in the study. RESULTS: Discrepancies between clinical and surgico-pathological staging occurred in 62% of cases. The tendency towards overdiagnosis was found to be increasing with local advancement of neoplasm. CONCLUSIONS: Accuracy of preliminary cervical cancer staging needs further improvement. PMID- 17076201 TI - [The influence of ovarian tumour histopathological diagnosis on prognostic values of the sonomorphological index]. AB - OBJECTIVES: The purpose of the study was to evaluate the helpfulness of sonomorphological index in preoperative diagnosis of ovarian tumours. MATERIAL AND METHOD: 686 patients diagnosed and treated because of adnexal tumours were analyzed. At the admission day the patient underwent an ultrasonographical examination that stated morphology of tumour using the sonomorphological index (SM). RESULTS: The sonomorphological index demonstrated high prognostic values: accuracy - 80.6%; sensivity - 86.7%; specifity - 77.0%. During the study, histopathological types of ovarian tumours with the lowest prognostic values were appointed. Among malignant tumours the lowest sensitivity of the test corresponded with: dysgerminoma and folliculoma. Among benign tumours the lowest helpfulness in preoperative diagnosis of ovarian tumours the sonomorphological index demonstrated in following histopathological types: mucinouse cystadenoma and adult teratoma. Particular histopathological diagnosis is the tubo-ovarian abscess--all analyzed cases had a false positive result in the test. CONCLUSIONS: In all histopathological types of malignant and benign tumours mentioned above, the use of the sonomorphological index require caution. To increase accuracy of preoperative diagnosis based on ultrasonographical examination, additional Doppler examination and also biochemical markers concentration should be considered. PMID- 17076202 TI - [Prevalence of Leptotrichia amnionii sp. nov. in pregnant women]. AB - INTRODUCTION: Leptotrichia bacteria belong to the group of Gram-negative anaerobes most frequently colonizing the oral cavity and reproductive organs. Leptotrichia sanquigenes constitute one of the bacterial factors in postpartum bacteremia of mothers and newborns, whilst Leptotrichia amnionii sp. nov., described for the first time in 2002, has turned out to be the etiopathogenetic factor in missed abortion in the second trimester of pregnancy. AIM: The purpose of this study was evaluation of the frequency of occurrence of Leptotrichia amnionii sp. nov. in patients in the 1st, 2nd and 3rd trimester of pregnancy. MATERIALS AND METHODS: The study involved 69 patients. Amplification reactions detecting the presence of Leptotrichia amnionii so. RNA in the smears were performed using specific starters complementary to 16SrRNA Leptotrichia amnionii- primer 1 Lam upper and primer 2 Lsp lower--complementary to Leptotrichia 16SrRNA. After amplification, the specimens were placed on 1% agarose gel in the presence of MassRuler marker (Fermentas). Next, PCR products were ligated to pGEM-T Easy Victor and then competent E. coli DH5 alpha cells were transformed with pGEM vector with an insert. Plasmid DNA isolation was performed using Plasmid Mini kit from A&A Biotechnology. Sequencing of inserts from the purified plasmids was performer in Molecular Biology Techniques Laboratory in the Department of Biology at Adam Mickiewicz University in Poznan. RESULTS: In the group of 69 pregnant patients Leptotrichia amnionii sp. nov. was identified in 8 women, which is 11.6%. CONCLUSION: Recently identified Leptorichia amnionii sp. nov. is a bacterie, which relatively frequently occurs in pregnant women. PMID- 17076203 TI - [Biophysical and biochemical intrapartum fetal assessment in relation to hematological varaiables in umbilical cord blood]. AB - The analysis of the hematological variables of umbilical cord blood can be useful in broadening our knowledge of both physiological and pathological processes undergoing during prenatal period. OBJECTIVES: The aim of our study was to evaluate the hematological parameters of umbilical cord blood in cases of either the abnormal fetal heart rate or the confirmed fetal acidosis. MATERIAL AND METHODS: A study group consisted of 458 neonates, live-born term of 38 to 41 completed weeks of single gestation. In each case, the hematological cord blood parameters were assessed. Fetal acidosis was diagnosed when the umbilical cord blood pH was below 7,15. Fetal heart rate (FHR) was assessed according to FIGO criteria. Statistical analysis was conducted using Mann-Withney tests, analysis of variance, p-value < 0.05 was considered as significant. RESULTS: The values of WBC (16,5 vs. 13,8x109), NRBC (25,1 vs. 7,3 /100 WBC) and HCT (51,8 vs. 46,5%) were higher in cases of fetus acidosis compare to non-acidemic fetuses. The significant differences in RBC, PLT, NRBC, HCT, HG were observed between group with abnormal and normal FHR. CONCLUSIONS: Fetal distress resulted in significant changes of the hematological variables in umbilical cord blood. Fetal hemopoetic system responds in specific manner to asphyxia. A precise analysis of fetal hematological variables may help to establish an exact onset and degree of fetal hypoxia. PMID- 17076205 TI - Repetitive demand for radical cancer risk reduction surgery in a young BRCA1 mutation carrier with strong family history of BRCA linked malignancies. AB - It is known that BRCA genes play central roles in hereditary breast and ovarian cancers. BRCA1 mutation carriers face a cumulative lifetime risk of ovarian and breast cancer development. We report on a case of a strong family prevalence of BRCA1 linked malignancies as an immense psychological encumbrance and reason of demand for radical prophylactic risk decreasing surgeries in a 29 year-old healthy woman with proved 3889delAG BRCA1 gene mutation on exon 11, codon 1265 in effort to prevent possible malignant changes in the ovaries and the breast. Problems regarding the management of asymptomatic BRCA mutation carriers, time and impact of early prophylactic surgery in young women are discussed with a review of recent literature. PMID- 17076204 TI - [Heaviness of the work measured by energy expenditure during pregnancy and its effect on birth weight]. AB - OBJECTIVES: Maternal work during pregnancy, especially with high work-related physical exertion, is still considered one of the most prevalent risk factors of negative pregnancy outcome. MATERIALS AND METHODS: Nofer Institute of Occupational Medicine has recently completed a study of women working in polish greenhouses. The greenhouse is a very good environment to measure the energy expenditure during work. That's why the women working for a period of at least 2 years in greenhouses in Poland were asked to participate in the project. We classified pregnancies of women on the basis of energy expenditure during mother's work into three groups: light work (200-700 kcal/shift); moderate work (701-1000 kcal/shift); and heavy work (1001-1200 kcal/shift). RESULTS: The study among women working in greenhouse revealed that the mean birth weight of infants whose mothers worked in greenhouse with heavy wok (work expenditure >1000 kcal/shift) during pregnancy was 177 g. lower, than that of those whose mothers worked out of greenhouse with light work (light work <700 kcal/shift) (p=0.05). CONCLUSIONS: The results suggest the necessity to inform pregnant women about the potential negative influence of heavy work on pregnancy outcome. PMID- 17076206 TI - [Laparoscopic sentinel lymph nodes identification followed by trachelectomy in early stage cervical cancer]. AB - The status of regional lymph nodes is the main prognostic factor in diagnosis and treatment of early stage of the cervical cancer. If the first node that drains a tumor site (sentinel node) is not metastatic, other lymph nodes should also be free of the disease. Detection using blue dye and laparoscopic removal of the sentinel lymph nodes let to avoid radical hysterectomy with pelvic limphadenectomy and it is especially useful in young women who want to preserve fertility. We describe a case of a 33- year old woman with invasive cervical cancer who underwent laparoscopic sentinel lymph nodes detection followed by trachelectomy. Thus histopatological examination confirmed no metastasis in removed sentinel lymph nodes, trachelectomy as a minimally invasive procedure was performed. PMID- 17076207 TI - [Successful termination of pregnancy complicated by fetal ascites--case report and review of the literature]. AB - This is a case of 25-year-old woman, primigravida, admitted to our department in 27th week of pregnancy complicated by fetal ascites revealed by ultrasound examination. The patient underwent diagnostic amniopunction and viral examinations in 20th week of pregnancy. The results from these studies were normal. This pregnant woman was qualified for fetal abdomen puncture, fluid evacuation and shunt applying between fetal abdomen and amniotic sac. Further outcome of pregnancy was uncomplicated. A live female child was born by caesarean section in 38th week of gestation. The shunt was removed after delivery. The woman and the baby were discharged in a good general condition. PMID- 17076208 TI - [Compliance endogenous and exogenous estriol in clinical practice]. AB - The aim of this study is presenting contemporary datas of the chemical structure, clinical meanings and the role of the weakest estrogen-estriol. Measurement of the level of estriol can serve as a marker of fetus's well-being, can detect some congenital diseases like: Down syndrome, trisomy 18. Inappropriate values of E3 are often associated with high risk pregnancies like: pregnancy-induced hypertension, miscarriage, preterm delivery or intrahepatic cholestasis. It seems, that estriol can be used in hormonal replacement therapy. Some experiments on animals indicates, that estriol administration can be effective in the treatment some autoimmunological diseases. PMID- 17076209 TI - [Catamenial pneumothorax]. AB - Catamenial pneumothorax (CP) is a specific type of secondary spontaneous pneumothorax. It is characterized by a temporal relationship with menses. CP is considered to be the most frequent symptom of thoracic endometriosis. It occurs mainly in women above 30 years of age and is typically right sided. The authors reviewed the current literature on this entity. The hypotheses on its pathogenesis and the diagnostic possibilities and therapeutic options are discussed. PMID- 17076210 TI - Putting the management of severe malnutrition back on the international health agenda. PMID- 17076211 TI - A review of methods to detect cases of severely malnourished children in the community for their admission into community-based therapeutic care programs. AB - BACKGROUND: The complexity and cost of measuring weight-for-height make it unsuitable for use by community-based volunteers. This has led community therapeutic care programs to adopt a two-stage screening and admission procedure in which mid-upper-arm circumference (MUAC) is used for referral and weight-for height is used for admission. Such a procedure results in many individuals being referred for care on the basis of MUAC but subsequently being refused treatment because they do not meet the weight-for-height admission criterion. This "problem of rejected referrals" has proved to be a major barrier to program uptake. OBJECTIVE: To systematically review methods to detect cases of severely malnourished children in the community for their admission into community-based therapeutic care programs. METHODS: Clinical and anthropometric methods for case detection of severely malnourished children in the community were reviewed with regard to their ability to reflect both mortality risk and nutritional status. RESULTS: MUAC, with the addition of the presence of bipedal edema, was found to be the indicator best suited to screening and case detection of malnutrition in the community. The case definition "MUAC < 110 mm OR the presence of bipedal edema," with MUAC measured by a color-banded strap, is suitable for screening and case detection of malnutrition in the community for children aged between 6 and 59 months. Monitoring and discharge criteria were also reviewed. CONCLUSIONS: There is no compelling evidence to support a move away from using weight in combination with clinical criteria for monitoring and discharge. PMID- 17076212 TI - Efficacy and effectiveness of community-based treatment of severe malnutrition. AB - BACKGROUND: There is a long tradition of community-based rehabilitation for treatment of severe malnutrition: the question is whether it is effective and whether it should be advised for routine health systems. OBJECTIVE: To examine the effectiveness of rehabilitating severely malnourished children in the community in nonemergency situations. METHODS: A literature search was conducted of community-based rehabilitation programs delivered by day-care nutrition centers, residential nutrition centers, primary health clinics, and domiciliary care with or without provision of food, for the period 1980-2005. Effectiveness was defined as mortality of less than 5% and an average weight gain of at least 5 g/kg/day. RESULTS: Thirty-three studies of community-based rehabilitation were examined and summarized. Eleven (33%) programs were considered effective. Of the sub-sample of programs reported since 1995, 8 of 13 (62%) were effective. None of the programs operating within routine health systems without external assistance was effective. CONCLUSIONS: With careful planning and resources, all four delivery systems can be effective. It is unlikely that a single delivery system would suit all situations worldwide. The choice of a system depends on local factors. High energy intakes (> 150 kcal/kg/day), high protein intakes (4-6 g/kg/day), and provision of micronutrients are essential for success. When done well, rehabilitation at home with family foods is more cost-effective than inpatient care, but the cost effectiveness of ready-to-use therapeutic foods (RUTF) versus family foods has not been studied. Where children have access to a functioning primary health-care system and can be monitored, the rehabilitation phase of treatment of severe malnutrition should take place in the community rather than in the hospital but only if caregivers can make energy- and protein dense food mixtures or are given RUTF. For routine health services, the cost of RUTF, logistics of procurement and distribution, and sustainability need to be carefully considered. PMID- 17076213 TI - Key issues in the success of community-based management of severe malnutrition. AB - BACKGROUND: Acute malnutrition is an underlying factor in almost 50% of the 10 to 11 million children under 5 years of age who die each year of preventable causes. Inpatient treatment for severe acute malnutrition is associated with high opportunity and economic costs for affected families and health service providers. Community-based therapeutic care attempts to address these problems and to maximize population-level impact through improving coverage, access, and cost-effectiveness of treatment. THE COMMUNITY-BASED THERAPEUTIC CARE MODEL: Community-based therapeutic care programs provide effective care to the majority of acutely malnourished people as outpatients, using techniques of community mobilization to engage the affected population and maximize coverage and compliance. People with severe acute malnutrition without medical complications are treated in an outpatient therapeutic program with ready-to-use therapeutic food and routine medication. Those suffering from severe acute malnutrition with medical complications are treated in an inpatient stabilization center according to standard World Health Organization protocols until they are well enough to be transferred to the outpatient therapeutic program. IMPACT OF COMMUNITY-BASED THERAPEUTIC CARE PROGRAMS: Twenty-one (21) community-based therapeutic care programs were implemented in Malawi, Ethiopia, and North and South Sudan between 2000 and 2005. These programs, which treated 23,511 cases of severe acute malnutrition, achieved recovery rates of 79.4% and mortality rates of 4.1%. Coverage rates were approximately 73%. Of the severely malnourished children who presented, 76% were treated solely as outpatients. Initial data indicate that these programs are affordable, with the cost-effectiveness of emergency community based therapeutic programs varying from US$12 to US$132 per year of life gained. PMID- 17076214 TI - Local production and provision of ready-to-use therapeutic food (RUTF) spread for the treatment of severe childhood malnutrition. AB - BACKGROUND: Ready-to-use therapeutic food (RUTF) spread has been shown to be very effective in the rehabilitation of severely malnourished children and facilitates home-based therapy of these children. OBJECTIVE: To describe how RUTF spread can be manufactured on a variety of production scales. PRINCIPLES OF PRODUCTION: RUTF spread is an edible lipid-based paste that is energy dense, resists bacterial contamination, and requires no cooking. The primary production principles include grinding all ingredients to a particle size < 200 microns, producing the food without the introduction of water, and embedding the protein and carbohydrate components of the food into the lipid matrix. The most widely used RUTF spread is a mixture of milk powder, sugar, vegetable oil, peanut butter, vitamins, and minerals. SCALE OF PRODUCTION: RUTF spread can be produced in quantities sufficient to treat several hundred children using a planetary mixer in a clinic. Production of larger quantities of RUTF spread can be achieved in partnership with local food companies. Production sufficient to meet the needs of several thousand children can be achieved with a dedicated production facility using technology appropriate for use in the developing world. Care must be taken to avoid aflatoxin contamination, and quality control testing of the product is essential. CONCLUSIONS: RUTF spread can be safely and easily produced in small or large quantities in most settings worldwide. The local availability of the necessary ingredients limits its use in some settings, and further investigation of alternative ingredients is needed to overcome this limitation. PMID- 17076215 TI - The sustainability of community-based therapeutic care (CTC) in nonemergency contexts. AB - BACKGROUND: Concern Worldwide is an international humanitarian nongovernmental organization that piloted and is now implementing and researching community-based therapeutic care (CTC) approaches to managing acute malnutrition. Experience in several countries suggests that there are key issues to be addressed at the international, national, regional, and community levels for community-based treatment of acute malnutrition to be sustainable. NATIONAL LEVEL: At the national level there must be demonstrated commitment to a clear health policy and strategy to address outpatient treatment of acute malnutrition. In addition, locally available, affordable ready-to-use therapeutic food (RUTF) must be accessible. REGIONAL LEVEL: At the regional level a functional health system and appropriate capacity for service provision are required. Integration of outpatient services should be viewed as a process with different levels of inputs at different phases depending on the capacity of the Ministry of Health (MOH). There is a need for indicators to facilitate scale-up and scale-back for future emergency response. COMMUNITY LEVEL: Strong community participation and active screening linked to health service provision at the local level is paramount for sustainable assessment and referral of severe acute malnutrition. FUTURE CHALLENGES TO SUSTAIN COMMUNITY-BASED THERAPEUTIC CARE. Key challenges to the sustainable treatment of severe acute malnutrition include the development of locally produced RUTF, development of international standards on local RUTF production, the integration of outpatient treatment protocols into international health and nutrition guidelines, and further operational research into integration of community-based treatment of severe acute malnutrition into health systems in nonemergency contexts. PMID- 17076216 TI - Proceedings of the WHO, UNICEF, and SCN Informal Consultation on Community-Based Management of Severe Malnutrition in Children. PMID- 17076217 TI - [Essential hypertension: pathogenetic studies continue]. PMID- 17076218 TI - [Dynamics of electrophysiological parameters of the heart in hypertensive patients depending on a 24-h profile of arterial pressure, left ventricular geometry and metabolic disorders]. AB - AIM: To examine conduction system and repolarization in the ventricles and heart rate variability in hypertensive patients with consideration of 24-h blood pressure profile, left ventricular (LV) geometry and metabolic disorders. MATERIAL AND METHODS: 24-h monitoring of blood pressure, diagnostic transesophageal electrostimulation of the left ventricle, echocardiography were made and duration and dispersion of QT interval, variability of the intervals R-R (SDNN) were assessed in 73 untreated patients aged 42 to 57 years with essential hypertension of the second degree. RESULTS: It is shown that hypertensive patients having left ventricular hypertrophy (LVH), metabolic syndrome (MS) and pathologic 24-h blood pressure profile have also a depressed function of the sinus-atrial node and atrioventricular conduction, marked electric instability of the atria and ventricles. Such patients are at high risk to develop arrhythmia (3 5 times higher than patients without LVH, MS, with normal circadian blood pressure rhythm). CONCLUSION: Electric heart remodeling associated with LVH, MS and disturbances of circadian blood pressure pattern enhances electric instability and risk to develop cardiac arrhythmia. PMID- 17076219 TI - [Effects of hostility on the risk of arterial hypertension, myocardial infarction, stroke in a sample of males aged 25-64 years (epidemiological trial on the base of WHO program MONICA)]. AB - AIM: To study effects of hostility on the risk of arterial hypertension (AH), myocardial infarction (MI) and stroke in males aged 25-64 years. MATERIALS AND METHODS: A screening study of the population was performed in 1994 according to WHO program MONICA-psychosocial. The response in a random representative sample of 25-64-year-old males (n = 657) living in Novosibirsk was 82.1%. New cases of MI, AH and stroke were registered in the control periods 1994-2000, 1994-2002. Computer program package SPSS-10 was used for statistic processing. Cox regression model of the relative risk (RR) was employed. RESULTS: Hostility was rather prevalent among the examinees (76.9%). MI risk for 8 years was 4.65 times higher in hostile men. Hostility was not associated with higher risk of AH and stroke. Hostility was seen more frequently in men with poor education, workers and retired persons. These groups are also at the highest risk of MI. CONCLUSION: Hostility raises MI risk in unstable society but had no effect on the risk of AH and stroke. PMID- 17076220 TI - [The degree of coronary artery calcinosis as a prognostic factor of asymptomatic cardiovascular complications: results of meta-analysis]. AB - AIM: To investigate prognostic implications of the calcium index estimated tomographycally for identification of patients at high risk to develop cardiovascular complications (non-fatal myocardial infarction, lethal coronary hear disease, coronary revascularization) in the absence of clinical manifestations. MATERIAL AND METHODS: Meta-analysis of the results of studies on prognostic implications of coronary arteries calcinosis was made with standardized risk ratio as a metameter. The meta-analysis included tests for systemic error related to publishing primarily positive results and data heterogenicity. RESULTS: Overall risk ratios for cardiovascular complications ranged from 2.1 to 9.3 for different degrees of coronary artery calcinosis. CONCLUSION: Calcium index is an independent risk factor of complications of cardiovascular diseases. PMID- 17076221 TI - [Pilot experience in Russia with external contrapulsation in patients with coronary heart disease]. AB - AIM: To examine effects of programmed external contrapulsation (ECP) on the clinico-functional status, quality of life (QOL) in patients with coronary heart disease (CHD), stable angina of FC II-IV resistant to medication. MATERIAL AND METHODS: Eighteen patients with CHD, stable angina of FC III-IV (2 females and 16 males, mean age 63.6 +/- 7.4 years) have completed a course of ECP including 35 one-hour procedures 5-6 times a week for 7 weeks. Ten patients had cardiac failure (NYHA FC II-III). Before and after ECP course the patients were examined using QOL assessment by Minnesota questionnaire, complex echo-cardiography, bicycle exercise test, perfusion myocardial scintigraphy with 99m-Tc-4,2-methoxy isobutilisonitril, 24-h Holter ECG monitoring, enzyme immunoassay for plasma natriuretic propeptides. RESULTS: Significant subjective QOL improvement (p < 0.01) was noticed by all the patients. Anginal attacks and nitrates doses reduced at least 2-fold. Exercise tolerance rose significantly (p < 0.01), bicycle exercise test was positive in 5 patients, the rest stopped the test after achievement of submaximal heart rate (HR) and fatigue. Most of the patients exhibited improvement of myocardial perfusion. Patients with abnormal myocardial contractility showed a moderate trend (p < 0.03) to an increase in left ventricular ejection fraction. By 24-h ECG, mean HR diminished significantly (p < 0.02). CONCLUSION: ECP is highly effective and safe in combined therapy of CHD, stable angina resistant to drug therapy, in impossibility of myocardial revascularization, including patients with cardiac failure. This manifests in a significant abatement of angina, lower doses of nitrates, improvement of exercise tolerance, quality of life, myocardial perfusion and hemodynamic indices. PMID- 17076222 TI - [Exercise in outpatient complex rehabilitation and secondary prophylaxis in patients with ischemic heart disease after acute coronary events (a cooperative trial in Russia)]. AB - AIM: To assess efficacy of early and long-term exercise in patients with ischemic heart disease (IHD) after acute coronary events (acute myocardial infarction- AMI, unstable angina--UA, coronary artery bypass grafting--CABG) in wide outpatient medical practice of Russia. MATERIAL AND METHODS: Three to eight weeks after the acute coronary event (ACE) 373 patients from different clinics of RF regions were randomized into two groups: the study group (n = 188) and a control one (n = 185). Both groups received standard therapy, the study group performed also a special exercise program (moderate exercise for maximum 1 hour 3 times a week for a year). The efficacy of the treatment was assessed by clinical, device and biochemical findings during 6-month follow-up. RESULTS: Physical work capacity, total amount of performed work (PW) in stress test rose in the study group by 26.5% (p < 0.01) and 59.4% (p < 0.01), respectively; HRmax and BPSmax x HRmax increased by 6.4 and 9.8%, respectively, p < 0.01); PW/Hrpeak rose by 48.6% (p < 0.01), PW/DP peak--by 45.9% (p < 0.01). Left ventricular ejection fraction increased by 5.6% (p < 0.05), LV stroke volume--by 3.4% (p < 0.05). In 6 months, controls demonstrated a 4.7% (p < 0.05) rise in HDLP cholesterol, total cholesterol lowered in the study group by 6.4% (p < 0.01), body mass index--by 1.9%, number of anginal attacks--by 0.9%. CONCLUSION: The proposed exercise program is effective and can be applied in outpatients after acute coronary events. The program can be also used as a method of secondary prophylaxis as it lowers total cholesterol, atherogenic index and body mass index. PMID- 17076223 TI - [MRT assessment of metabolic and thrombolytic therapy effects on postinfarction left ventricular remodeling]. AB - AIM: To examine effects of trimetasidine on morphofunctional indices of the left ventricle (LV) in myocardial infarction (MI) patients on combined treatment. MATERIALS AND METHODS: Seventy five patients with primary macrofocal MI were randomized into 2 groups. Patients of group 1 (n = 38) received a combination of bisoprolol (beta-blocker) with enalapril (ACE inhibitor) in individual doses under control of blood pressure and blood creatinine level. Group 2 (n = 37) patients received the same combination of drugs and, in addition, trimetasidine in a dose 70 mg/day from postmyocardial day 7-10 for 6 months. Two subgroups from the groups were given systemic thrombolytic therapy (STLT) with streptokinase. MRT and cine-MRT of the heart were made for measurement of LV morphofunctional parameters. RESULTS: Low-field MRT of the heart in MI patients treated with adjuvant STLT (1500000 U within 6 hours after MI onset) and trimetasidine (preductal MB) in a dose 70 mg/day from the disease day 7-10 registered a significant inhibition of pathological LV postinfarction remodeling: a decrease of body surface indexed LV end diastolic volume by 10.3%, systolic volume--by 15.4%, LV myocardial tension--by 14.0%, sphericity index--by 7.1%; an increase in the index of relative wall thickness by 5.3%, cardiac index--by 9.2% compared to the group treated without trimetasidine. CONCLUSION: 6-month therapy with trimetasidine of MI patients leads to a significant regress of morphofunctional changes accompanying LV remodeling. Pathological LV postinfarction remodeling inhibits significantly in MI patients combined treatment of whom included STLT (streptokinase in a dose 1500,000 U within 6 hours after acute MI onset and trimetasidine in a dose 70 mg/day on postmyocardial infarction day 7-10). PMID- 17076224 TI - [Cardioselective beta-adrenoblockers in patients with stable angina pectoris. Comparison of efficacy and safety]. AB - AIM: To compare efficacy and safety of cardioselectivc beta-blockers (BB) in patients with stable angina. MATERIAL AND METHODS: An open comparative randomized trial with participation of 40 patients suffering from stable angina (NYHA FC II III) was made. All the patients were divided into two groups. Patients of group 1 received betaxolol, those of group 2--other selective BB: metoprolol (n = 13), bisoprolol N = 6, nebivolol (n = 1). The BB dose was doubled each 2 weeks up to achievement of the maximal tolerable dose. The trial continued for 2 months during which the patients were also given aspirin, statins, ACE inhibitors, mononitrates, amlodipin and thiazide diuretics. The efficacy of the treatment was assessed by the data of the treadmill test by Brus protocol, echocardiography according to ASE recommendations, high resolution ultrasound examination of endothelium-related relaxation of the brachial artery. Lipid transport parameters were estimated with enzyme techniques. RESULTS: A mean dose of betaxolol, metoprolol and bisoprolol was 14 +/- 4.5 mg/day, 127 +/- 24 mg/ day, 10 +/- 4 mg/day. A noticeable lowering of heart rate (HR) and blood pressure (BP) was observed in both groups, but HR in group 1 decreased more (57.35 +/- 5.19 and 62.4 +/- 8.84 b/m, respectively, p = 0.033) and systolic pressure showed a trend to greater reduction. Exercise tolerance in both groups was compatible. The lowest threshold HR was achieved in betaxolol group (a fall from 133.8 +/- 23.5) to 105.0 +/- 14, 23 b/min (p+0.027). Endotheium-related relaxation of the brachial artery was improved in betaxolol group: the diameter of the artery increased from 6.38 +/- 4.32 to 9.22 +/- 4.37% (p = 0.057), the peak blood flow velocity--from 14.81 +/- 3.91 to 23.87 +/- 3.7% (p = 0.031). In group 2 a positive trend in these parameters was not observed. BB had no negative effect on left ventricular contractility, parameters of transmitral blood flow, bronchial conduction, metabolism. CONCLUSION: Compared to other BB, betaxolol had a stronger effect on hemodynamic parameters (HR and BP) at rest and exercise, improved endothelial vascular function in patients with stable angina. PMID- 17076225 TI - [Amiodaron effects on atrioventricular conduction]. AB - AIM: To assess short- and long-term effects of amiodaron on atrioventricular conduction (AVC) of the heart. MATERIAL AND METHODS: Amiodaron was given to 477 patients (mean age 48.7 +/- 0.7 years) with various arrhythmias caused, as a rule by coronary heart disease. A mean saturation dose was 809.4 +/- 13.4 mg/day, a mean maintanence dose--263.5 +/- 5.4 mg/day. Mean follow-up 20.85 +/- 1.2 months. The frequency of AVC disorders was estimated at regular Holter monitoring. RESULTS: In saturation, atrioventricular block of the first degree arose in 17.2% patients, of the second and third degree--only in one patient. AVC disorders disappeared at reduction of amiodaron dose. Only in one patient the drug was discontinued because of recurrent block of the third degree and bradycardia. CONCLUSION: In saturation, ECG records lengthening of P-R intervals by 18.9%, on the average. Maintenance therapy increases the length of P-R interval by 8.1%, on the average. PMID- 17076226 TI - [Immune status of patients with cardiac arrhythmias: idiopathic and in primary heart disease]. AB - AIM: A detailed description of immune status abnormalities of adult patients with heart arrhythmia either idiopathic or in combination with primary heart disease such as chronic myocarditis and dilated cardiomyopathy (DCMP). MATERIAL AND METHODS: Eighty two consecutive patients aged 16-57 years admitted to the L.A. Myasnikov Institute of Clinical Cardiology (Moscow) for heart arrhythmia were studied. Among them 35 patients had idiopathic heart arrhythmia (IHA, group 1) with no evidence of any primary heart disease, while other 47 patients (group 2) had heart arrhythmia combined with primary heart disease (chronic myocarditis or DCMP). In group 1 ventricular arrhythmia was recorded in 27 patients (12 cases with ventricular tachyarrhrythmia ?VTA, 15 cases with ventricular extrasystolia- VE). Supraventricular heart arrhythmia was found in 6 patients (3 cases of constantly recurring supraventriccular tachycardia, 2 cases of paroxysmal and 1 with constant atrial fibrillation). The intermittent atrioventricular block of the second-third degree was recorded in 2 patients. The patients of group 2 were divided into subgroups 2a, 2b and 2c. In subgroup 2a (patients with DCMP without signs of heart failure) ventricular arrhythmia was found in 7 patients (VT ? 5, VE ? 2). Supraventricular arrhythmia was recorded in 7 patients 5 of which had constantly recurring supraventricular tachycardia, 1 ? paroxysmal and 1 constant atrial fibrillation. In subgroup 2b (DCMP patients with obvious signs of heart failure) ventricular arrhythmia was recorded in 12 patients, among them 6 had VT and 6 ? VE, 2 ? constant atrial fibrillation). In subgroup 2c (patients with chronic myocarditis) ventricular arrhythmia was in 7 patients (VT ? 5, VE ? 2), constant atrial fibrillation ? in 2, heart conduction abnormalities ? in 3 patients, atrioventricular block of the first or second degree ? in 2, sick sinus syndrome ? in 1. To verify the diagnosis, all the patients have undergone physical examination, blood cell counts and biochemical tests, urine clinical analysis, ECG and ultrasound heart examination as well as 24h ECG monitoring. On demand, bicycle exercise test or treadmill test, coronaroangiography, endomyocardial biopsy and invasive electrophysiological examination were made. RESULTS: Immune status abnormalities found in patients with heart arrhythmia both idiopathic and combined with primary heart diseases such as chronic myocarditis and DCMO correspond to immune defense response during chronic infection. Activation of different anti-infection defense mechanisms was recorded in patients with idiopathic heart rhythm and conductivity abnormalities. Immune deficiency was found in arrhythmia and conductivity abnormalities combined with primary heart diseases (chronic myocarditis or DCMP). A positive correlation exists between the degree of immune defense failure and reduction of myocardial contractility. CONCLUSION: There exists a characteristic pattern of immune status abnormalities in patients with arrhythmia, both idiopathic or combined with primary heart disease (myocarditis, DCMP). The abnormalities depend on severity of arrhythmia, intensity of inflammatory processes in the myocardium and on the degree of left ventricular contractility dysfunction in patients with primary heart diseases. PMID- 17076227 TI - [Is triple combination of different neurohormonal modulators recommended for treatment of mild-to-moderate congestive heart failure patients? (Results of SADKO-CHF study). Part 2]. AB - AIM: To assess different variants of neurohormonal (NH) modulation with angiotensin converting enzyme (ACE-I) quinapril (Q), angiotensin-receptor blocker (ARB) valsartan (V) and their combination in addition to beta-adrenergic blocker bisoprolol (B) on functional status, quality of life (QL), parameters of left ventricular (LP) remodeling, main indices of 24-h heart rate variability (HRV) and NH profile in patients with stable mild-to-moderate CHF. MATERIAL AND METHODS: 63 patients with CHF (NYHA class II-III) as a result of ischemic heart disease and dilated cardiomyopathy with LV EF < 40% were randomly assigned to one of the treatment variants on 1:1:1 basis: B+Q (n = 22; mean daily dose of B-5.5 mg; Q-15.4 mg), B+V (n = 23; mean daily dose of B = 4.8 mg; V = 128 mg) and combination of B+Q+V (n = 18; mean daily dose of B = 4.1 mg; Q = 12 mg; V = 82 mg). At baseline, all the patients in this study were on background B treatment and according to the study design Q or V were then added to B at randomization. NYHA FC, 6-min walking test (6MT), QL, 2D-echocardiography, plasma rennin activity (PRA), angiotensin II (AT-II), aldosterone (Ald), norepinephrine (NE), epinephrine (E), brain natriuretic peptide (BNP) concentrations and 24-hour HRV parameters were investigated at baseline, 3 and 6 months after randomization. RESULTS: During the study NYHA FC improvement was revealed in all 3 treatment groups with comparative significant changes in 6MT distance by 20.4%, 19.1% and 19.4% in B+Q, B+V and B+Q+V groups. QL maximally decreased in B+V combination (from 45 to 21 points). LV volumes significantly decreased and LV ejection fraction (EF) increased in all groups to the end of the study. Triple combination had no additional effect on LV volumes and LVEF changes compared to B+Q and B+V groups. Maximally plasma NE concentrations decreased in B+Q group (from 650 to 430 pg/ml, p = 0.007). A worse effect was observed in the combination of B+Q+V, with any NE changes in B+V group. The E concentration increased significantly (from 215 to 295 pg/ml, p = 0.024) in the B+Q+V group at the end of the study. Plasma A-H concentration did not differ from the baseline during the study in B+Q group, but significantly increased in B+V group and maximally in B+Q+V group (from 11.4 to 23.5 pg/ml, p = 0.009). To the end of the study plasma Ald concentrations remain reduced significantly only in B+V group. The level of BNP significantly decreased in all 3 treatment groups. Significant changes in HRV indices, both in time and frequency domain, were revealed in the B+Q group at 3 month follow-up and SDNN increased on month 24 (p = 0.039). These changes became insignificant at the end of the study. The lesser effect was revealed in B+Q+V group, with insignificant trend toward an increase of SDNN to the end of the study. HRV indices did not improve in the B+V group. CONCLUSION: During long-term treatment the triple combination of B+Q+V has no significant advantages over B+Q and B+V by the functional status, QL and parameters of LV remodeling in patients with mild-to-moderate CHF. The combination of B+Q has more potent effect on 24 hour HRV parameters, sympatho-adrenal activity and renal function compared to B+V and B+Q+V groups in CHF patients in our study. The combination B+Q+V may have a negative effect on NH profile (excessive activation of ATII and E) in CHF patients. The triple combination is not recommended for therapy of stable mild-to moderate CHF patients. PMID- 17076228 TI - [Basic results of the multicenter trial NOKTURN]. AB - AIM: Integration of noliprel treatment of arterial hypertension with educational programs on correction of associated risk factors in outpatient practice for reduction of the overall cardiovascular risk. MATERIAL AND METHODS: An open, multicenter, non-comparative trial with participation of 140 physicians from 14 regions of Russia comprised 1195 patients aged 30-65 years with mild-to-moderate hypertension (SBP = 140-179 mmHg, and/or DBP = 90-109 mm Hg) who had never received regular antihypertensive therapy. This one-year study had an active 3 month stage and follow-up stage for 9 months. RESULTS: Noliprel significantly decreased both systolic and diastolic blood pressure. The response to the treatment was achieved in 95.6% patients, the target blood pressure was achieved in 76.2% patients. A mean level of total cholesterol lowered from 5.5 +/- 1.0 mmol/l to 5.1 +/- 0.8 mmol/l (p < 0.001), body mass index--from 28.4 +/- 4.5 to 27.2 +/- 4.3 (p < 0.001). Hypercholesterolemia occurred before the study in 65.5% patients, at the end of the study--in 49.2%, obesity was registered in 79 and 70.2%, smoking in 21.6 and 14.6%, respectively. The mean overall risk to develop fatal cardiovascular events reduced from 2.5 +/- 3.6% at initiation of the study to 1.3 +/- 1.8% in the end of the study (p < 0.001). CONCLUSION: The integration of medication with noliprel and training program successfully lowered overall risk to develop fatal outcomes of cardiovascular diseases in primary health care service. PMID- 17076229 TI - [Mildronate in geriatric patients with cardiac failure]. AB - AIM: To study efficacy of mildronate in the treatment of elderly patients with cardiac failure (CF). MATERIAL AND METHODS: A total of 91 patients with chronic CF (NYHA FC I-III) entered the study. The study group of 63 patients received standard therapy for a month and mildronate in a dose 750 mg/day. The control group of 28 patients was treated conventionally. Subjective and objective assessment of the patients' condition, quality of life questionnaire, ECG, 6-min walk test were used for evaluation of the treatment efficacy. RESULTS: In the study group daily frequency of anginal attacks reduced from 1.6 to 0.7 (in the control group from 1.46 to 1.25, p > 0.05), intensity of the attacks--from 1.4 to 0.7 points by 7-score scale (p < 0.05). Crepitation disappeared in 8 (12.8%) patients, edema of the legs--in 3 (4.8%) patients. Systolic blood pressure went down by 8 mmHg, diastolic one--by 4 mmHg. In the control group clinical changes were insignificant. CONCLUSION: Elderly patients tolerated mildronate well. It was a safe drug attenuating CF symptoms, increasing exercise tolerance and improving quality of life. PMID- 17076230 TI - [Ivabradin (coraxan). One more new approach to fighting myocardial ischemia in patients with ischemic heart disease]. PMID- 17076231 TI - [Remodeling of the heart after transcatheter repair of the secondary interatrial defect with amplatzer occluder]. AB - AIM: To ascertain the effects of transcatheter repair of the secondary interatrial defect (IAD) with Amplatzer occluder on heart remodeling in respect to the patient's age. MATERIAL AND METHODS: Transcatheter repair of the secondary IAD was made in 64 patients. Two groups were formed by the patients' age: group 1 -age under 16 years (mean age 9 +/- 0.6 years) and group 2--age over 16 years (mean age 32 +/- 1.8 years). The control group consisted of 30 healthy subjects matched by age. Echocardiography measured size and volumes of the heart chambers, intracardiac hemodynamics before treatment of the defect and 24 hours, 1, 3, 6, 12 and 24 months after it. RESULTS: The indexed diameter of the defect in group 1 was 16.5 +/- 1.6 mm/m2, in group 2--13.7 +/- 0.7 mm/m2 (p > 0.05). The size of the occluder, in relation to the surface area, was 20 +/- 1.5 and 16 +/- 0.8 mm/m2, respectively (p < 0.05). 24 hours after the defect repair the right ventricle volume reduced by 29% in group 1 and by 21%--in group 2. The right atrium volume reduced significantly also. Mean pulmonary arterial pressure (PAP) in both groups diminished from 22 +/- 0.7 to 19 +/- 0.7 mmHg and from 24 +/- 1.0 to 20 +/- 0,8 mmHg, respectively. The left atrium decreased by 34 and 32%, respectively (p < 0.001). Remodeling reduced RV-LV in both groups by 22 and 23% (p < 0.001), respectively. 3 months after the defect repair all the indices reached control values. 12 months after the correction hemodynamics in the patients was the same with the healthy examinees. The time and degree of reduction of the right heart were similar in the groups, the volumes diminished by about 50%. 12 months after the repair in 4 of 18 patients of group 2 (22%) the right atrium and ventricle were larger than in healthy subjects though less than before the repair. CONCLUSION: Transcatheter repair of the secondary IAD with Amplatzer catheter leads to early noticeable reduction of the right heart. The type of the changes is similar in both age groups and does not depend on the size of the implanted occluder. Not all the patients over 16 years of age achieve normal sizes of the RV and RA 12 months after the defect correction. PMID- 17076232 TI - [The problem of indexation of left ventricular myocardial mass by body size in obese patients]. AB - AIM: To assess efficacy of different ECG-criteria of left ventricular myocardial hypertrophy (LVH) in hypertensive patients as regards body mass (obesity). MATERIAL AND METHOD: Data on 100 patients (42 males and 58 females, age 19-79 years) with diagnosis of hypertension of the first-second degree were analysed. ECG was registered in 12 leads. LVH was determined by ECG by the following criteria: Sokolov-Lion (S-L): Sv1+Rv5(v6) > 35 mm; Cornell voltage (Crn-V): R avL+Sv3 > 28 mm (> 20 mm for women); Cornell product (Crn-P). According to the body mass the patients were divided into 3 groups: with normal weight, overweight and obesity. Crn = P criterion was most sensitive in all the three groups. RESULTS: In the groups sensitivity of ECG criteria depends on several factors: on the method of indexation and on the gender and body mass index. CONCLUSION: Informative value of ECG criteria of LVH depends on the method of indexation and on the amount of the excessive body mass. PMID- 17076233 TI - [Effects of fertilizing levels on output and quality of Pinellia ternate]. AB - OBJECTIVE: To compare the effects of the different fertilizing levels on the output, the effective element, the content of heavy-metal, the grade and the ratio of wet to dry of Pinella ternate to obtain the best fertilize ratio on the planting. METHODS: The orthogonal desing method L9 (3(4)) was applied on the comparison between the different fertilization levels. The content of alkaloid was determined by UV spectrophotometer. The content of heavy-metal was determined by atom absorption. RESULTS: The factors that affected the output and the quality of Pinellia ternate were N > P > K. CONCLUSION: The best fertilizer combination is N 25 g/m2 ,P2O5 18 g/m2, K2O9 g/m2 and the application ratio is 2. 8: 2: 1 for them, in which the higher output and the content of alkaloid should be obtained. PMID- 17076234 TI - [Purification and activity calculated of fibrinolyric factors from Eupolyphaga sinensis]. AB - Three kinds ( EFF-1, EFF-2 and EFF-3) of fibrinolytic factor were separated by ammonium sulphate precipitation, DEAE-cellulose and preparative PAGE electrophoresis from female Eupolyphaga sinensis Walker. Their molecular weights were proved to be 41kd, 32.9 kd and 30.6 kd respectively with SDS-PAGE electophoresis. Their Activities as plasminogen activator were 171.3 U/mg, 234.0 U/mg and 148.5 U/mg. In addition, EFF-2 and EFF-3 were not only fibrinolytic activities but also have plasminogen activator on fibrinous plate lacked of plasminogen . There had been no such components of plasminogen activator and fiberinolytic enzyme from Eupolyphaga sinensis reported yet. PMID- 17076235 TI - [The extraction and identification of total flavanone from longan pericarp]. AB - OBJECTIVE: In order to make use of the resources of total flavanone in pericarp of longan. METHODS: The flavonids were extracted with ethanol. Used spectrophotometry to check the flavanone. RESULTS: The total flavanone in the pericarp was 1.101 mg/ml and the rate of recovery was 99.98%. CONCLUSION: The extraction and purifying methods in this experiment can get the outcome and the purity of the flavanone very high. This method is a purely physical process and has not any pollution. It is an ideal way to extract the flavanone of longan skin. PMID- 17076236 TI - [Studies on chemical constituents of Serissa serissoides roots]. AB - OBJECTIVE: To study the chemical constituents from Serissa serissoides DC. roots. METHODS: Seven compounds were obtained from the 80% ethanol extract of Serissa serissoides roots by repeated column chromatography over silica gel, Sephadex LH 20 and Polyamide. Their structures were identified by analysis of their spectral data and physics character. RESULTS: Seven compounds obtained from the plant leaves were identified as: (+)-pinoresinol(1), (-)-syringaresinol( 2), (+) meduiresinol(3), (-)-olivil(4), beta-sitosterol(5), oleanolic acid(6) and daucosterol(7). CONCLUSION: They are all known compounds. Compound 2 and 4 was obtained from this genus plants for the first time, and compounds 1,3 and 7 as known compounds were obtained from the root of the plant firstly. PMID- 17076237 TI - [Analysis of the essential oils from Amomun kravak Pierre ex Gagnep by GC-MS]. AB - OBJECTIVE: To establish the fingerprint chromatogram of Amomun kravak Pierre ex Gagnep. METHODS: The composition of the essential oil was isolated by PR solvent and determined by GC-MS. Forty-two species of compounds were identified, of which the relative contents were calculated using square peaks to normalization, among which the character components were 1,8-cineole, beta-pinene, alpha-pinene, p cymene, etc. EMIC software was used and EMIC fingerprint chromatogram was established by selecting the character ion of those 10 compounds. Different extraction methods were also compared. RESULTS: The GC-MS fingerprint chromatogram can provide qualitative information confirming compounds of the corresponding chromatographic peaks. EMIC was first used to reason the quality control of the medicine quality and it can establish the foundation for safe and effective quality control of the medicine and its product. PR solvent can easily extract the essential oil from the medicine. CONCLUSION: This method has the advantage of high efficiency and low cost, and the GC-MS fingerprint chromatogram provided the ascription and quality of the medicine and will play an important role in promoting the modernization of Chinese traditional medicine. PMID- 17076238 TI - [Content comparison of pinoresinol diglucoside in original and reborn bark of Eucommia ulmoides]. AB - OBJECTIVE: To evaluate the reborn barks quality, antihypertensive effective component of pinoresnol diglucoside (PDG) in three times reborn barks were determined. METHODS: A YWG C18 column (10 microm, 250 x 4.6 mm) was used with a mobile phase of methanol-water (30:75) and flow rate of 1.0 (ml/min). The detective wave-length was set at 277nm and the column temperature at room temperature. RESULTS: PDG in the first reborn barks are slightly higher than the original ones, and in the second reborn barks are similar with the barks before girdling (the fist reborn barks), but in the third reborn barks, PDG are much lower than the barks before girdling (the second reborn barks). CONCLUSION: In order to ensure reborn barks quality, we suggest that the girdling bark regeneration can be made two times, the time between the first and the second girdling is not less than five years. PDG in the third reborn barks should be enhanced. PMID- 17076239 TI - [Experimental study on the effect of tiangui gengnian capsule on the aged female rats ostepoprosis]. AB - OBJECTIVE: To study the therapeutic action of Seabuckthorn Fatty Acid on the aged female rats osteoporosis, and explore the corresponding mechanism related in hormone and cytokine. METHODS: The change of bone density (BMD), biomechanics and bone morphology were measured and E2, TGF-beta1, IGF-1 level in plasma were determined respectively for the young female rats, aged female rats (Seabuckthorn Fatty Acid 0.72, 1.8, 4.5 g/kg, E2 0.02 g/kg) after they were administrated corresponding drugs for 45 days. RESULTS: The bone density (BMD), bone biomechanics, bone morphology for female rats aged 22-months all decreased. The BMD, bone biomechanics, the number of bone trabeculas and cortical thickness in Seabuckthorn Fatty Acid treated groups (1.8 g/kg, 4.5 g/kg) were upgraded (P < 0.05). The TGF-beta1, IGF-1 level in plasma were increased, too (P < 0.05). CONCLUSION: Seabuckthorn Fatty Acid can meliorate bone metabolism in post menopause and heighten the bone density. The adjustment mechanism may be based on the function of estrogen and cell cytokine. PMID- 17076240 TI - [Pharmacokinetics and tissue distribution of atractylenolide III in rats]. AB - OBJECTIVE: [corrected] To establish an HPLC method for the analysis of pharmacokinetics and tissue distribution of atractylenolide III in rats. METHODS: The biological samples were extracted with ether. The chromatographic conditions were as follows: Hypersil ODS column (150 mm x 4.6 mm, 5 microm) was used. The mobile phase was methnol/warter (67 : 33) with a flow rate of 1.0 ml/min under the column temperature of 25 degrees C, and the detection wavelength was set at 220 nm. RESULTS: The recovery of the method was 85.12% (RSD = 5.57%). The linear range was 0.2 microg/ml - 18.5 microg/ml (r = 0.9996) in rat plasma. The Lowest Limit of detection was 0.10 microg/ ml (S/N > 3). The within-day and between-day precision were from 0.98% to 6.19% and 12.95% to 15.48%, respectively. After oral administration of atractylenolide III (100 mg/kg), the concentration-time profiles of atractylenonlide III fit a two compartment model. In main effect tissues, the atractylenolide III concentration was followed as in order C(lung) > C(cerebellum) > C(heart) > C(cerebrum), and that was C(spleen) > C(liver) > C(kidney) in eliminated tissues. CONCLUSION: The method is accurate, stable and reliable, and can be used for the investigation of atractylenolide III in plasma and tissues of rats. PMID- 17076241 TI - [Protective effect of effective parts of Zingiber Offecinal on vascular endothelium of the experimental hyperlipidemic rats]. AB - OBJECTIVE: To observe the influence of effective parts of Zingiber Officinale on serum IL-6, TNF-alpha in oroler to investigate the protective effects of the effective parts of Zingiber Officinal (EPZ) on endothelium of the experimental hyperlipidemic rats and the mechanism of its effects. METHODS: The hyperlipidemia model of rats was constructed by feeding high-fat forage and filled with the effective parts of Zingiber Officinale 200 mg/kg, 400 mg/kg, 800 mg/kg every day for 13 weeks. Blood was drawn to determine both the level of serum IL-6 and TNF alpha. All the aortaes were taken to oberserve morphologic change and the intima media thickness were detected. RESULTS: The effective parts of Zingiber Officinale could markedly decrease intima-media thickness, but had no marked influence in the level of serum IL-6 and TNF-alpha. CONCLUSION: The Effect Parts of Zingiber Officinale has the effect of protection of the endothelia of hyperlipidemia rats, which has nothing with the level of serum IL-6 and TNF alpha. PMID- 17076242 TI - [Effect of ligustrazine, synthetic borneol on lasting time of ciliary movement]. AB - OBJECTIVE: To study the using of the method in vitro and in vivo of lasting time of ciliary movement in ligustrazine, synthetic borneol. METHODS: (1) Method in vitro-The lasting time of ciliary movement was observed with forty-fold optical microscope after dropping liquid medicine on exteacorporeal frog palate mucosa. Then mucosa was cleared. It was observed whether the ciliary movement was recovered and the lasting time was recorded from recovering to stopping once again. (2) Method in vivo-Liquid medicine or normal saline was dropped on frog palate mucosa by contacting thirty minutes. Then mucosa was cleared. The lasting time of ciliary movement was observed with forty-fold optical microscope by separating palate mucosa. RESULTS: Relatively percentage of lasting time of ciliary movement of ligustrazine in vitro and in vivo was 9.8%, 87.3%; The relatively percentage of synthetic borneol in vitro and in vivo was 9.3%, 89.5%. CONCLUSION: The method in vitro and in vivo of lasting time of ciliary movement can be the one of the selecting ways of Chinese drug's toxicity of nasal mucosa, and it have virtues and defects respectively. Ligustrazine, synthetic borneol have significantly toxical effect on exteacorporeal lasting time of ciliary movement. PMID- 17076243 TI - [Studies on the effect of extracts of several Chinese herbal medicines and other medicines on alcohol dehydrogenase activity]. AB - OBJECTIVE: To study the effects of water and alcohol extracts of several Chinese herbal medicines and other medicines on alcohol dehydrogenase activity in order to provide enzymology basis on new medicine. METHODS: Water or alcohol extracts of Chinese herbal medicine and other medicine were tested on the effects of alcohol dehydrogenase activity by Valle and Hoch method. RESULTS: Among them, 8 were found to have the effect of activation on alcohol dehydrogenase. They were water extracts of Amomum kravanh and Pueraria flowers, the alcohol extracts of Pueraria flowers, compound hepatcare Chinese medicine and compound Pueraria medicine, L-cysteine, notoginseng saponin. Others had inhibiting action. CONCLUSION: To decrease alcohol concentration in the body through activating the activity of ADH may be one of the mechanisms for some traditional Chinese herbal medicine in neutralizing the effect of alcohol drink. PMID- 17076244 TI - [Preparation of puerarin solid self-microemulsion]. AB - OBJECTIVE: To prepare puerarin solid self-microemulsifying drug delivery system (SMEDDS). METHODS: Pseudo-ternary phase diagram was used to select the formula of microemulsion. The self-microemulsification efficiency was assessed, such as viscosity, conductance, refraction, zeta potential, size ditribution, dissolution were investigated. Puerarin were determined by UV to calculate the cumulative release at different time. RESULTS: The optimum formulation of puerarin solid self-microemulsifying drug delivery system the consisted of puerarin-Tween 800 glycerinum-ethyl oleate-water-Gum Acacia-manicol at weight ratio of 1.7: 16. 6: 16. 6: 16. 6 : 1.3: 4.0: 39.9: 19.9; Average particle diameter was 30 nm; The dissolution was 94.29% at 10 min. CONCLUSION: Stability of puerarin solid self microemulsion drug delvery system is good, potent in improving the dissolution of puerarin. PMID- 17076245 TI - [Studies on flavonoids extraction technology from Glycyrrhiza inflata and their bacteriostatic activities]. AB - OBJECTIVE: To study the extraction technique for the flavonoids from Glycyrrhiza inflata and their bacteriostatic bioactivities. METHODS: It was investigated about comparison of extraction methods such as ultrasonic-assisted extraction (UAE), enzymatic extraction (EE), whisk extraction (WE) and repeat freeze dissolve extraction (FDE). The UAE conditions were optimized by orthogonal array. The bacteriostatic effect was studied using doubling dilution. RESULTS: The UAE was the most effective method and the optimal UAE conditions were adding 20-fold ethanol (30% v/v), extracting for two times, 20 min each time. Futhermore, the minimal inhibitory concentrations (MIC) of the flavonoids against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Bacillus subtilis were 0.03125, 0.03125, 0.125, 0.03125 mg/ml respectively. CONCLUSION: UAE is suitable for fast and effective extraction of flavonoids. The flavonoids have obvious bacteriostatic bioactivities against microorganisms mentioned above. PMID- 17076246 TI - [Effect of micronization on the quality of Salvia miltiorrhiza Bge]. AB - OBJECTIVE: The powder characteristic, water extraction amount and active ingredients cryptotanshinone (CTS), tanshinone II A (TS) and protocaechuic aldehyde (PA) were comparatively studies between crude powder and micronized powder to study the application of micronization technology to Salvia miltiorrhiza Bge. METHODS: Salvia miltiorrhiza Bge powedr was characterized by laser diffraction analyzer and scanning electron microscopy. The angle of repose and bulk density was measured. The water extraction was quantified by heated extraction method. The active ingredinents CTS, TS and PA were detected by RP HPLC after it released from Salvia miltiorrhiza Bge. RESULTS: The differences of particle characteristic and surface modality between crude powder and superfine were significant. Water extraction amount of micronized powder was 1.23 times as that of crude powder. The CTS, TS and PA extraction amount of micronized powder were 1.54, 1.48 and 1.90 times as those of crude powder. CONCLUSION: The extraction content of lipo-solubility and water-solubility active ingredients of Salvia miltiorrhiza Bge. are improved by micronization. Micronization can economize the resource of Salviae miltiorrhiza Bge. The bioavability of Salviae miltiorrhiza Bge. will be improved with the new techonology. PMID- 17076247 TI - [HPLC fingerprint of Danggui Buxue decoction]. AB - OBJECTIVE: To establish the fingerprint of soup of Danggui Buxue decoction. METHODS: HPLC with Nucleodur C18 Gravity colum was used and the Acetonitrile water (gradient elution) as a mobile phase and detecting wavelength at 203 nm. RESULTS: There were 15 main peaks in the soup of Danggui Buxue decoction. 15 come from Radix astragali and 7 come from Radix angelicae sinensis. CONCLUSION: This fingerprint can be used as a reference for stablility of soup of Danggui Buxue decoction. PMID- 17076248 TI - [Primary study on the SPE-HPLC fingerprint of Gaoli ginseng injection]. AB - OBJECTIVE: To study on the fingerprint of Gaoli ginseng Injection. METHODS: SPE was used to purify samples. HPLC with DAD detector was used to analysis the samples of Gaoli ginseng Injection. Then the analogical degree of the chromatography was estimated by the software. RESULTS: The fingerprints had 14 communal chromatsogram peaks, and the analogical degrees of all samples were greater than 0.95. CONCLUSION: The method is credible. It provides a scientific basis to control the quality of Gaoli ginseng Injection more effectively. PMID- 17076249 TI - Expressed sequence tags from feline uterine library. AB - Partial nucleotide sequences of 634 cDNAs randomly isolated from a feline uterine cDNA library (Stratagene) were determined by single pass sequencing. Homology search of the sequences to the non-redundant nucleotide databases revealed that 83% of the cDNAs matched registered feline or non-feline genes. Based on the gene identifications, these genes were predicted to be related with immunological, biochemical and regulatory functions in cats. Interestingly, the rest 17% of the cDNAs did not show homology to gene or EST sequence present in the nucleotide and protein databases, suggesting that these cDNAs include novel genes expressed only in the Felidae. This large scale sequencing of uterine cDNA will provide a useful molecular source for research not only towards health and disease conditions in cats but also in different fields of science where genetic information from cats will be of interest. PMID- 17076250 TI - Sequence analysis of UTR and coding region of kappa-casein gene of Indian riverine buffalo (Bubalus bubalis). AB - In this study, complete nucleotide as well as derived amino acid sequence characterization of water buffalo (Bubalus bubalis) kappa-casein gene has been presented. Kappa-casein cDNA clones were identified and isolated from a buffalo lactating mammary gland cDNA library. Sequence analysis of kappa-casein cDNA revealed 850 nucleotides with an open reading frame (ORF) of 573 nucleotides, encoding mature peptide of 169 amino acids. The 5' untranslated region (UTR) comprised 71 nucleotides, while 3' UTR was of 206 nucleotides. A total of 11 nucleotide and seven amino acid changes were observed in, buffalo (Bubalus bubalis) as compared to cattle (Bos taurus), sheep (Ovis aries) and goat (Capra hircus). Among these nucleotide changes, eight were unique in buffalo as they were fully conserved in cattle, sheep and goat. Majority of the nucleotide changes and all the amino acid changes; 14 (Asp-Glu), 19(Asp/Ser-Asn), 96(Ala Thr), 126(Ala-Val), 128(Ala/Gly-Val), 156(Ala/Pro-Val) and 168(Ala/Glu-Val) were limited to exon IV. Three glycosylation sites, Thr 131, Thr 133 and Thr 142 reported in cattle and goat kappa-casein gene were also conserved in buffalo, however, in sheep Thr 142 was replaced by Ala. Chymosin hydrolysis site, between amino acids Phe 105 and Met 106, important for rennet coagulation process, were found to be conserved across four bovid species. Buffalo kappa-casein with the presence of amino acids Thr 136 and Ala 148 seems to be an intermediate of "A" and "B" variants of cattle. Comparison with other livestock species revealed buffalo kappa-casein sharing maximum nucleotide (95.5%) and amino acid (92.6%) similarity with cattle, whereas with pig it showed least sequence similarity of 76.0% and 53.2%, respectively. Phylogenetic analysis based on both nucleotide and amino acid sequence indicated buffalo kappa-casein grouping with cattle, while sheep and goat forming a separate cluster close to them. The non-ruminant species viz. camel, horse and pig were distantly placed, in separate lineages. PMID- 17076251 TI - The mitochondrial genome of Cygnus columbianus, the Whistling Swan. AB - The nucleotide sequence of the mitochondrial genome of the Whistling Swan, Cygnus columbianus, is reported. Many of the features common to avian mitochondrial genomes are present in C. columbianus and are described here. The gene order is the same as in Gallus gallus. The sequence of this mitochondrial genome allows relationships within the family Anatidae (swans, geese and ducks) to be reconsidered in the light of a large suite of mitochondrial characters. Protein coding gene sequences of C. columbianus were concatenated to form a supergene, which was analyzed phylogenetically with similar constructs from previously published avian genomes. Relationships within Anatidae and between the Anatidae and the galliform birds were addressed. Three independent phylogenetic methods confirmed traditional classifications and the existence of the Galloanseres clade. PMID- 17076252 TI - Strict conservation of the ITS regions of the ribosomal RNA genes in Atlantic cod (Gadus morhua L.). AB - The nucleotide sequence of the internal transcribed region (ITS) of ribosomal RNA genes from Atlantic cod (Gadus morhua L.) was determined. The complete ITS region spanned approximately 1113 base pairs. The ITS1 region comprised 532 base pairs, the 5.8S region 159, and the ITS2 region contained 422 base pairs. Sequence data were obtained from a total of 12 samples, one pool from six cod and 11 individuals. The sequencing was carried out in two separate experimental periods employing slightly different methodology. The samples were from two different cod stocks, Norwegian costal cod and North East Arctic cod. The sequence analysis showed that in the 12 samples, the ITS region, including the 5.8S RNA, was identical. The ITS region is thus totally conserved in these two cod stocks. The extreme conservation of the ITS regions in the cod rDNA could reflect the small genome size of cod and/or indicate a specific critical role in the processing of the ribosomal units in cold-adapted species. PMID- 17076253 TI - The complete nucleotide sequence of the mitochondrial genome of Tetraodon nigroviridis. AB - The fresh water pufferfish Tetraodon nigroviridis is a model organism for studying evolution of genome and gene functions, but its mitochondrial genome (mtDNA) sequence is still not available. We determined the complete nucleotide sequence of its mtDNA using shotgun sequencing. The T. nigroviridis mtDNA was 16,462 bp, and contained 13 protein coding genes, 22 tRNAs, 2 rRNAs and a major non-coding region. The gene order was identical to the common type of vertebrate mtDNA, whereas the G + C content in the sense strand was 46.9%, much higher than most other fish species. One hundred and three SNPs were detected in the control region of the mtDNA of 35 individuals, a majority of SNPs were detected in the 5' end of the control region. A phylogenetic study including 21 fish species was performed on concatenated amino acid sequences of 12 protein coding genes, and revealed that the T. nigroviridis was clustered with Fugu rubripes into a group. The complete mtDNA sequence and SNPs in its control region will be useful in studying fish evolution, in differentiating different Tetraodon species and in analyzing genetic diversity within T. nigroviridis. PMID- 17076254 TI - Identification and tissue-specific expression of amphioxus GM2 activator protein gene from amphioxus Branchiostoma belcheri. AB - An amphioxus cDNA, AmphiGM2AP, encoding GM2 activator protein was isolated from the gut cDNA library of Branchiostoma belcheri. It is 907 bp long, and its longest open reading frame codes for a precursor protein consisting of 242 amino acid residues with a signal peptide of 14 amino acids. The deduced amino acid sequence includes a conserved domain typical of GM2APs between residues 53 and 224, a single N-linked glycosylation site at position 65 and 8 conserved cysteines. Phylogenetic analysis showed that amphiGM2AP forms a club together with invertebrate GM2APs, indicating that AmphiGM2AP is evolutionarily closely related to invertebrate GM2APs rather than vertebrate ones. Both Northern blotting and in situ hybridization histochemistry analyses revealed a tissue specific expression pattern of AmphiGM2AP in adult amphioxus with the strongest expression in the digestive system, which is in contrast to the widespread expression pattern of human, mouse and sheep GM2AP genes. It is suggested that AmphiGM2AP is possibly involved in the take-in of digested food components like lipid molecules. PMID- 17076255 TI - Cloning, expression and characterization of phycoerythrin gene from Ceramium boydenn. AB - Phycobiliproteins function as a major light harvesting protein-pigment complex in the cyanobacteria and the eukaryotic algae. Phycoerythrin (PE) is a kind of phycobiliproteins, widely located in all rhodophytes, some species of cyanobacteria and cryptophytes, and different ecotypes of Prochlorococcus populations. PeBA encoding beta and alpha subunits of PE from Ceramium boydenn was cloned and sequenced in this research. A peBA specific PCR primer was synthesized, based on the peBA gene conserved sequences. The beta subunit encoding gene (peB) contained an open reading frame of 534 bp, while the alpha subunit (peA) was 495 bp. Recombinant expression plasmid pET-peAB was constructed and expressed in Escherichia coli BL21. The molecular weight of expressive product of peB and peA was about 23.3 and 18.2 KD, respectively. Results of codon usage analysis show that G + C content is heterogeneous among different groups of PE and spacers have dramatically lower G + C contents than coding regions. Also there is a high variance in G + C content among sequences at the third position sites. It is also found in this paper that several sequence regions, which might reflect functional or structural requirements of the PE organization, and several residues known for their functional importance are conserved in almost all the sequences. PMID- 17076256 TI - Cloning and characterization of a cDNA encoding Ran binding protein from wheat. AB - Ran, which functions in nucleocytoplasmic transport and mitosis, binds to and is regulated in part by Ran binding protein (RanBP). A RanBP cDNA (TaRanBP1) was isolated from a wheat cDNA library using RT-PCR product as a probe. The predicted amino acid sequence of TaRanBP1 is over 60% identity to AtRanBP1 from Arabidopsis and also with considerable similarity to human and fungi RanBPs. TaRanBP1 gene was expressed ubiquitously in roots, leaves and stems, with a similar abundance in these tissues. Phylogenetic reconstruction of TaRanBP1 with 32 other RanBPs from 26 species of organisms revealed that RanBPs from plants, animals and fungi clustered as the distinct groups, intraspecies isoforms were not developed for RanBPs, contrast with most other ancestral genes. Structural analysis revealed that all RanBPs were highly conserved in the middle region of their amino acid sequence, which included Ran binding domain and the three conserved motifs that have the essential roles in binding with Ran protein and promotion of GTP hydrolysis by the Ran/RanGAP/RanBP complex. However, N-terminus and C-terminus exhibited very low similarity between the different RanBPs. The different structures in N-terminus and C-terminus of RanBPs are likely to direct the Ran into the specific physiological processes and subsequently exhibit the different roles in different organisms. PMID- 17076257 TI - Cloning and characterization of a bamboo LEAFY HULL STERILE1 homologous gene. AB - A cDNA named DlMADS8 was isolated from the young spikelets of the sweet bamboo, Dendrocalamus latiflorus by rapid amplification of cDNA end (RACE). DNA sequence analysis showed that DlMADS8 was composed of full ORF and 3'UTR, but without 5'UTR. The cDNA contained 1059 nucleotides and encoded a putative protein of 244 amino acid residues. The gene displayed the structure of a typical plant MADS-box gene, which consisted of a MADS domain, K domain, a short I region, and the C terminal region. Phylogenetic analysis of plant MADS-box genes based on amino acid sequences revealed that DlMADS8 was grouped into the AGAMOUS-LIKE 2 (AGL2) like subfamily. It was homologous to the LEAFY HULL STERILE1 (LHS1) genes of grasses. To study the functions of it, DlMADS8 cDNA clone driven by the CaMV 35S promoter was transformed into Arabidopsis thaliana. Transgenic plants of DlMADS8 exhibited the phenotypes of curled leaves and early flowering. After bolting, three novel phenotypes related to inflorescence development were observed in different transgenic plants. No obvious homeotic conversions of floral organs were observed in all of the 35S::DllMADS8 transgenic Arabidopsis plants. These results indicated that DlMADS8 probably plays a role in floral meristem determinacy and is involved in controlling the flowering time of D. latiflorus. PMID- 17076258 TI - Molecular cloning and expression analysis of rice OsTVLP1, encoding a protein with similarity to TGF-beta receptor interacting proteins and vacuolar assembly Vam6p/Vps39p proteins. AB - We describe the cloning and identification of a rice cDNA, OsTVLP1, encoding a protein with similarity to TGF-beta receptor interacting proteins and vacuolar assembly Vam6p/Vps39p proteins. OsTVLP1 has an open reading frame of 2955 bp, which encodes a 984 amino acid protein, containing a citron homology (CNH) domain at its N-terminal and a clathrin heavy-chain repeat homology (CLH) domain at its C-terminal. The expression of OsTVLP1 was induced by treatments with benzothiadiazole (BTH), a chemical activator of plant disease resistance responses, and by infection of the blast fungus, Magnaporthe grisea. Importantly, the expression of OsTVLP1 was activated specifically in disease resistance response induced by BTH and in an incompatible interaction between rice and the blast fungus. Our observations suggest that OsTVLP1 may play a role in rice disease resistance response against pathogen infection. PMID- 17076259 TI - Cloning and expression analysis of a hsp70 gene from Saussurea medusa. AB - Members of the 70 kD heat-shock gene family are highly conserved across a wide range of organisms. In an effort to learn more about the evolution and possible functions of extreme environment plant Saussurea medusa Maxim hsp70, we isolated a cDNA clone encoding a putative cytosolic member (Smhsp70) of this family of proteins from a cDNA library of S. medusa cell cultures. The cDNA clone was 2224 bp in length and contained a 1941 bp open reading frame (ORF) encoding a polypeptide of 647 amino acid residues with a predicted molecular mass of 70,794 Da. The predicted protein was found to contain a C-terminal amino acid motif of "PKIEEVD" indicating that Smhsp70 was related to cytosolic members of the hsp70 family in higher plant. The secondary and three-dimensional structures of Smhsp70 were analyzed by molecular modeling. The genomic structure of Smhsp70 included one intron of 1134 bp in length. The deduced Smhsp70 protein has 93.7 and 93.2% similarity with the hsp70 of tobacco and tomato, 73.2% with the hsp70 of human, and 43.7% with DnaK of Escherichia. coli, respectively. Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) analyses indicated that the cytosolic Smhsp70 protein was constitutively expressed and markedly increased after relatively short periods of heat shock (37 degrees C) as well as by low temperature (4 degree C) treatments. PMID- 17076260 TI - Using universal degenerate primers for restriction digestion mapping by PCR. AB - In this study, a polymerase chain reaction (PCR) is developed to determine the restriction map without using restriction endonucleases. A 937 bp fragment of pUC 19 which contained one cut site for EcoRI and two recognition sites for PvuII was used as a model. The PCR was carried out using designed degenerate primers and the products were analyzed on 1.5% agarose gel. The number of cut sites, length of fragments and the arrangement of the fragments from 3' or 5' end of desired sequence were determined. PMID- 17076261 TI - Natural antisense transcripts of Alzheimer's disease associated genes. AB - Natural antisense transcripts (NATs), also named endogenous antisense transcripts, are a class of genes whose role in controlling gene expression is becoming more and more relevant. NATs might play important roles in gene expression and translation regulation. Present work investigated the presence of NATs of Alzheimer's disease associated genes including PRESENILIN1, PRESENILIN2, BACE1, BACE2, APP, APOE, TAU (MAPT), PRION, alpha-SYNUCLEIN (SNCA), NICASTRIN, PEN2, APH1A, APH1B as well as CD147 (BASIGIN), and the results revealed that APP, BACE2, APH1A, TAU, CD147 and alpha-SYNUCLEIN contain natural antisense transcripts. These NATs were characterized according to the sense-antisense overlapping information and potential functional mechanisms were proposed. Present findings provide preliminary but important information about transcription regulation of AD associated genes, which would further our understanding of the gene expression regulation of AD, and also suggest a novel potential strategy for the therapy of AD. PMID- 17076262 TI - Methods and platforms for the quantification of splice variants' expression. AB - The relatively limited number of human protein encoding genes highlights the importance of the diversity generated at the level of the mRNA transcripts. As alternative RNA splicing plays a key role in mediating this diversity, it becomes critical to develop the tools and platforms that will deliver quantitative information on the specific expression levels associated with splice isoforms. This chapter describes the constraints generated by this global transcriptome analysis and the state-of-the-art techniques and products available to the scientific community. PMID- 17076263 TI - Pre-mRNA missplicing as a cause of human disease. AB - Regulated alternative splice site selection emerges as one of the most important mechanisms to control the expression of genetic information in humans. It is therefore not surprising that a growing number of diseases are either associated with or caused by changes in alternative splicing. These diseases can be caused by mutation in regulatory sequences of the pre-mRNA or by changes in the concentration of trans-acting factors. The pathological expression of mRNA isoforms can be treated by transferring nucleic acids derivatives into cells that interfere with sequence elements on the pre-mRNA, which results in the desired splice site selection. Recently, a growing number of low molecular weight drugs have been discovered that influence splice site selection in vivo. These findings prove the principle that diseases caused by missplicing events could eventually be cured. PMID- 17076264 TI - Alternative splicing: therapeutic target and tool. AB - Alternative splicing swells the coding capacity of the human genome, expanding the pharmacoproteome, the proteome that provides targets for therapy. Splicing, both constitutive and regulated forms, can itself be targeted by conventional and molecular therapies. This review focuses on splicing as a therapeutic target with a particular emphasis on molecular approaches. The review looks at the use of antisense oligonucleotides, which can be employed to promote skipping of constitutive exons, inhibit inappropriately activated exons, or stimulate exons weakened by mutations. Additionally this manuscript evaluates methods that reprogram RNAs using reactions that recombine RNA molecules in trans. Preliminary, but exciting, results in these areas of investigation suggest that these methods could eventually lead to treatments in heretofore intractable ailments. PMID- 17076266 TI - Misregulation of tau alternative splicing in neurodegeneration and dementia. AB - Tau is a microtubule-associated protein that fulfills several functions critical for neuronal formation and health. Tau discharges its functions by producing multiple isoforms via intricately regulated alternative splicing. These isoforms modulate tau function in normal brain by altering the domains of the protein, thereby influencing its conformation and post-translational modifications and hence its affinity for microtubules and other ligands. Disturbances in tau expression result in disruption of the neuronal cytoskeleton and formation of pathological tau structures (neurofibrillary tangles) found in brains of dementia sufferers. More specifically, aberrations in tau splicing regulation directly cause several neurodegenerative diseases that lead to dementia. This review briefly presents our cumulative knowledge of tau splicing regulation in connection with the alterations in tau splicing seen in neurodegeneration. PMID- 17076265 TI - SR proteins as potential targets for therapy. AB - Serine- and arginine-rich (SR) proteins constitute a highly conserved family of pre-mRNA splicing factors that play key roles in the regulation of splice site selection, and thereby in the control of alternative splicing processes. In addition to conserved sequences at the splice junctions, splice site selection also depends upon different sets of auxiliary cis regulatory elements known as exonic and intronic splicing enhancers (ESEs and ISEs) or exonic and intronic silencers (ESSs and ISSs). Specific binding of SR proteins to their cognate splicing enhancers as well as binding of splicing repressor to silencer sequences serve to enhance or inhibit recognition of weak splice sites by the splicing machinery. Given that the vast majority of human genes contain introns and that most pre-mRNAs containing multiple exons undergo alternative splicing, mutations disrupting or creating such auxiliary elements can result in aberrant splicing events at the origin of various human diseases. In the past few years, numerous studies have reported several approaches allowing correction of such aberrant splicing events by targeting either the mutated sequences or the splicing regulators whose binding is affected by the mutation. The aim of the present review is to highlight the different means by which it is possible to modulate the activity of SR splicing factors and to bring out those holding the greatest promises for the development of therapeutic treatments. PMID- 17076267 TI - Spinal muscular atrophy and therapeutic prospects. AB - The molecular genetic basis of spinal muscular atrophy (SMA), an autosomal recessive neuromuscular disorder, is the loss of function of the survival motor neuron gene (SMN1). The SMN2 gene, a nearly identical copy of SMN1, has been detected as a promising target for SMA therapy. Both genes are ubiquitously expressed and encode identical proteins, but markedly differ in their splicing patterns: While SMN1 produces full-length (FL)-SMN transcripts only, the majority of SMN2 transcripts lacks exon 7. Transcriptional SMN2 activation or modulation of its splicing pattern to increase FL-SMN levels is believed to be clinically beneficial and therefore a crucial challenge in SMA research. Drugs such as valproic acid, phenylbutyrate, sodium butyrate, M344 and SAHA that mainly act as histone deacetylase inhibitors can mediate both: they stimulate the SMN2 gene transcription and/or restore the splicing pattern, thereby elevating the levels of FL-SMN2 protein. Preliminary phase II clinical trials and individual experimental curative approaches SMA patients show promising results. However, phase III double-blind placebo controlled clinical trials have to finally prove the efficacy of these drugs. PMID- 17076269 TI - Redirecting splicing to address dystrophin mutations: molecular by-pass surgery. AB - Mutations in the dystrophin gene that prevent synthesis of a functional protein lead to Duchenne muscular dystrophy (DMD), the most common serious childhood muscular dystrophy. The major isoform is produced in skeletal muscle and the size of the dystrophin gene and complexity of expression have posed great challenges to the development of a therapy for DMD. Considerable progress has been made in the areas of gene and cell replacement, yet it appears that any potential therapy for DMD is still some years away. Other approaches are being considered, and one that has generated substantial interest over the last few years is induced exon skipping. Antisense oligonucleotides have been used to block abnormal splice sites and force pre-mRNA processing back to the normal patterns. This approach is re-interpreted to address the more common dystrophin mutations, where normal splice sites are targeted to induce abnormal splicing, resulting in specific exon exclusion. Selected exon removal during processing of the dystrophin pre-mRNA can by-pass nonsense mutations or restore a disrupted reading frame arising from genomic deletions or duplications. Attributes of the dystrophin gene that have hampered gene replacement therapy may be regarded as positive features for induced exon skipping, which may be regarded as a form of by-pass surgery at the molecular level. In humans, antisense oligonucleotides have been more generally applied to down-regulate specific gene expression, for the treatment of acquired conditions such as malignancies and viral infections. From interesting in vitro experiments several years ago, the dystrophin exon-skipping field has progressed to the stage of planning for clinical trials. PMID- 17076270 TI - Altered splicing in prelamin A-associated premature aging phenotypes. AB - Hutchinson-Gilford progeria (HGPS), a rare and severe developmental disorder characterized by features recalling premature aging, and restrictive dermopathy (RD), a neonatal lethal genodermatosis, have recently been identified as being primary or secondary "laminopathies." These are heterogeneous disorders due to altered function of lamins A/C or related proteins. In physiological conditions, mature lamin A is obtained through a series of post-translational processing steps performed on a protein precursor, prelamin A. The major pathophysiological mechanism involved in progeria is an aberrant splicing of pre-mRNAs issued from the LMNA gene, due to a de novo heterozygous point mutation, leading to the production and accumulation of truncated lamin A precursors. Aberrant splicing of prelamin A pre-mRNAs causing the production of more extensively truncated precursors is involved in the allelic disease restrictive dermopathy. Other restrictive dermopathy cases are due to the inactivation of a key enzyme involved in the maturation of lamin A precursors (ZMPSTE24). In functional terms, all these conditions share the same pathophysiological basis: intranuclear accumulation of lamin A precursors, which cannot be fully processed (due to primary or secondary events) and exert toxic, dominant negative effects on nuclear homeostasis. Most other laminopathies are due to autosomal dominant LMNA point mutations inferred to cause single amino acid substitutions. In any case, the impact of these mutations on pre-mRNA splicing has rarely been assessed. These disorders affect different tissues and organs, mainly including bone, skin, striated muscles, adipose tissue, vessels, and peripheral nerves in isolated or combined fashions, giving rise to syndromes whose severity ranges from mild to perinatally lethal. In this chapter we review the structure and functions of lamins A/C in physiological and pathological conditions, describe their known or putative roles, namely, in the pathogenesis of HGPS and RD in relation to existing animal models, and envisage possible targeted therapeutic strategies on the basis of recent research results. PMID- 17076268 TI - Misregulation of alternative splicing causes pathogenesis in myotonic dystrophy. AB - Myotonic dystrophy (DM), the most common form of adult onset muscular dystrophy, affects skeletal muscle, heart, and the central nervous system (CNS). Mortality results primarily from muscle wasting and cardiac arrhythmias. There are two forms of the disease: DM1 and DM2. DM1, which constitutes 98% of cases, is caused by a CTG expansion in the 3' untranslated region (UTR) of the DMPK gene. DM2 is caused by a CCTG expansion in the first intron of the ZNF9 gene. RNA containing CUG- or CCUG-expanded repeats are transcribed but are retained in the nucleus in foci. Disease pathogenesis results primarily from a gain of function of the expanded RNAs, which alter developmentally regulated alternative splicing as well as pathways of muscle differentiation. The toxic RNA has been implicated in sequestration of splicing regulators and transcription factors thereby causing specific symptoms of the disease. Here we review the proposed mechanisms for the toxic effects of the expanded repeats and discuss the molecular mechanisms of splicing misregulation and disease pathogenesis. PMID- 17076271 TI - Splicing modulation as a modifier of the CFTR function. AB - A significant fraction of CF-causing mutations affects pre-mRNA splicing. These mutations can generate both aberrant and correct transcripts, the level of which varies among different patients. An inverse correlation was found between this level and disease severity, suggesting a role for splicing regulation as a genetic modifier. Subsequent studies showed that overexpression of splicing factors modulated the level of correctly spliced RNA, transcribed from minigenes carrying CF-causing splicing mutations. Overexpression of splicing factors also modulated the level of normal CFTR transcripts, transcribed from the endogenous CFTR allele carrying splicing mutations, in CF-derived epithelial cells. Several of the factors promoted higher level of correct CFTR transcripts. The increased level of normal transcripts led to activation of the CFTR channel and restoration of its function. Restoration was also obtained by sodium butyrate, a histone deacetylase inhibitor, known to up-regulate the expression of splicing factors. These results highlight the role of the splicing machinery as a modifier of disease severity in patients carrying splicing mutations and shed a new light on the therapeutic potential of splicing modulation for genetic diseases caused by splicing mutations. PMID- 17076272 TI - Neural crest cells and the community of plan for craniofacial development: historical debates and current perspectives. AB - After their initial discovery in the mid 1800s, neural crest cells transitioned from the category of renegade intra-embryonic wanderers to achieve rebel status, provoked especially by the outrageous claim that they participate in skeletogenesis, an embryonic event theretofore reserved exclusively for mesoderm. Much of the 20th century found neural crest cells increasingly viewed as a unique population set apart from other embryonic populations and more often treated as orphans rather than fully embraced by mainstream developmental biology. Now frequently touted as a fourth germ layer, the neural crest has become a fundamental character for distinguishing craniates from other metazoans, and has radically redefined perceptions about the organization and evolution of the vertebrate jaws and head. In this chapter we provide an historical overview of four main research areas in which the neural crest have incited fervent discord among workers past and present. Specifically, we describe how discussions surrounding the neural crest threatened the germ layer theory, upended traditional schemes of vertebrate head organization, challenged assumptions about morphological conservation and homology, and redefined concepts on mechanisms of craniofacial patterning. In each case we frame these debates in the context of recent data on the developmental fate and roles of the neural crest. PMID- 17076273 TI - Neural crest inducing signals. AB - The formation of the neural crest has been traditionally considered a classic example of secondary induction, where signals form one tissue elicit a response in a competent responding tissue. Interactions of the neural plate with paraxial mesoderm or nonneural ectoderm can generate neural crest. Several signaling pathways converge at the border between neural and nonneural ectoderm where the neural crest will form. Among the molecules identified in this process are members of the BMP, Wnt, FGF and Notch signaling pathways. The concerted action of these signals and their downstream targets will define the identity of the neural crest. PMID- 17076274 TI - Transcriptional regulation at the neural plate border. PMID- 17076275 TI - Neural crest delamination and migration: integrating regulations of cell interactions, locomotion, survival and fate. AB - During the entire process of neural crest development from specification till final differentiation, delamination and migration are critical steps where nascent crest cells face multiple challenges: within a relatively short period of time that does not exceed several hours, they have to change drastically their cell- and substrate-adhesion properties, lose cell polarity and activate the locomotory machinery, while keeping proliferating, surviving and maintaining a pool of precursors in the neural epithelium. Then, as soon as they are released from the neural tube, neural crest cells have to adapt to a new, rapidly-changing environment and become able to interpret multiple cues which guide them to appropriate target sites and prevent them from distributing in aberrant locations. It appears from recent studies that, behind an apparent linearity and unity, neural crest development is subdivided into several independent steps, each being governed by a multiplicity of rules and referees. Here resides probably one of the main reasons of the success of neural crest cells to accomplish their task. PMID- 17076276 TI - Neural crest cell plasticity. size matters. AB - Patterning and morphogenesis of neural crest-derived tissues within a developing vertebrate embryo rely on a complex balance between signals acquired by neural crest cells in the neuroepithelium during their formation and signals from the tissues that the neural crest cells contact during their migration. Axial identity of hindbrain neural crest is controlled by a combinatorial pattern of Hox gene expression. Cellular interactions that pattern neural crest involve signals from the same key molecular families that regulate other aspects of patterning and morphogenesis within a developing embryo, namely the BMP, SHH and FGF pathways. The developmental program that regulates neural crest cell fate is both plastic and fixed. As a cohort of interacting cells, neural crest cells carry information that directs the axial pattern and species-specific morphology of the head and face. As individual cells, neural crest cells are responsive to signals from each other as well as from non-neural crest tissues in the environment. General rules and fundamental mechanisms have been important for the conservation of basic patterning of neural crest, but exceptions are notable and relevant. The key to furthering our understanding of important processes such as craniofacial development will require a better characterization of the molecular determinants of the endoderm, ectoderm and mesoderm and the effects that these molecules have on neural crest cell development. PMID- 17076277 TI - The contribution of the neural crest to the vertebrate body. AB - As a transitory structure providing adult tissues of the vertebrates with very diverse cell types, the neural crest (NC) has attracted for long the interest of developmental biologists and is still the subject of ongoing research in a variety of animal models. Here we review a number of data from in vivo cell tracing and in vitro single cell culture experiments, which gained new insights on the mechanisms of cell migration, proliferation and differentiation during NC ontogeny. We put emphasis on the role of Hox genes, morphogens and interactions with neighbouring tissues in specifying and patterning the skeletogenic NC cells in the head. We also include advances made towards characterizing multipotent stem cells in the early NC as well as in various NC derivatives in embryos and even in adult. PMID- 17076278 TI - Cranial neural crest and development of the head skeleton. AB - The skeletal derivatives of the cranial neural crest (CNC) are patterned through a combination of intrinsic differences between crest cells and extrinsic signals from adjacent tissues, including endoderm and ectoderm. In this chapter, we focus on how CNC cells positionally interpret these cues to generate such highly specialized structures as the jaw and ear ossicles. We highlight recent genetic studies of craniofacial development in zebrafish that have revealed new tissue interactions and show that the process of CNC development is highly conserved across the vertebrates. PMID- 17076279 TI - Neural crest contribution to the cardiovascular system. AB - Normal cardiovascular development requires complex remodeling of the outflow tract and pharyngeal arch arteries to create the separate pulmonic and systemic circulations. During remodeling, the outflow tract is septated to form the ascending aorta and the pulmonary trunk. The initially symmetrical pharyngeal arch arteries are remodeled to form the aortic arch, subclavian and carotid arteries. Remodeling is mediated by a population of neural crest cells arising between the mid-otic placode and somite four called the cardiac neural crest. Cardiac neural crest cells form smooth muscle and pericytes in the great arteries, and the neurons of cardiac innervation. In addition to the physical contribution of smooth muscle to the cardiovascular system, cardiac neural crest cells also provide signals required for the maintenance and differentiation of the other cell layers in the pharyngeal apparatus. Reciprocal signaling between the cardiac neural crest cells and cardiogenic mesoderm of the secondary heart field is required for elaboration of the conotruncus and disruption in this signaling results in primary myocardial dysfunction. Cardiovascular defects attributed to the cardiac neural crest cells may reflect either cell autonomous defects in the neural crest or defects in signaling between the neural crest and adjacent cell layers. PMID- 17076280 TI - The genetic regulation of pigment cell development. AB - Pigment cells in developing vertebrates are derived from a transient and pluripotent population of cells called neural crest. The neural crest delaminates from the developing neural tube and overlying ectoderm early in development. The pigment cells are the only derivative to migrate along the dorso-lateral pathway. As they migrate, the precursor pigment cell population differentiates and expands through proliferation and pro-survival processes, ultimately contributing to the coloration of organisms. The types of pigment cells that develop, timing of these processes, and final destination can vary between organisms. Studies from mice, chick, Xenopus, zebrafish, and medaka have led to the identification of many genes that regulate pigment cell development. These include several classes of proteins: transcription factors, transmembrane receptors, and extracellular ligands. This chapter discusses an overview of pigment cell development and the genes that regulate this important process. PMID- 17076281 TI - Specification of sensory neuron cell fate from the neural crest. AB - How distinct cell fates are generated from initially homogeneous cell populations is a driving question in developmental biology. The neural crest is one such cell population that is capable of producing an incredible array of derivatives. Cells as different in function and form as the pigment cells in the skin or the neurons and glia of the peripheral nervous system are all derived from neural crest. How do these cells choose to migrate along distinct routes, populate defined regions of the embryo and differentiate into specific cell types? This chapter focuses on the development of one particular neural crest derivative, sensory neurons, as a model for studying these questions of cell fate specification. In the head, sensory neurons reside in the trigeminal and epibranchial ganglia, while in the trunk they form the spinal or dorsal root ganglia (DRG). The development of the DRG will be the main focus of this review. The neurons and glia of the DRG derive from trunk neural crest cells that coalesce at the lateral edge of the spinal cord (Fig. 1). These neural crest cells migrate along the same routes as neural crest cells that populate the autonomic sympathetic ganglia located along the dorsal aorta. Somehow DRG precursors must make the decision to stop and adopt a sensory fate adjacent to the spinal cord rather than continuing on to become part of the autonomic ganglia. Moreover, once the DRG precursors aggregate in their final positions there are still a number of fate choices to be made. The mature DRG is composed of many neurons with different morphologies and distinct biochemical properties as well as glial cells that support these neurons. PMID- 17076282 TI - Neural crest and the development of the enteric nervous system. AB - The formation of the enteric nervous system (ENS) is a particularly interesting example of the migratory ability of the neural crest and of the complexity of structures to which neural crest cells contribute. The distance that neural crest cells migrate to colonize the entire length of the gastrointestinal tract exceeds that of any other neural crest cell population. Furthermore, this migration takes a long time--over 25% of the gestation period for mice and around 3 weeks in humans. After colonizing the gut, neural crest-derived cells within the gut wall then differentiate into glial cells plus many different types of neurons, and generate the most complex part of the peripheral nervous system. PMID- 17076283 TI - Growth factors regulating neural crest cell fate decisions. AB - Because of its unique ability to generate a wide variety of both neural and nonneural derivatives, the neural crest is an ideal model system to study the factors regulating cell lineage decisions in stem and progenitor cells. The use of various cell culture techniques and in vivo functional assays, including cell type-specific gene manipulation in mouse, helped to identify signaling factors involved in this process. Moreover, it became apparent that the biological functions of growth factors acting on neural crest cells depend on the context provided by the extracellular microenvironment. Thus, signaling molecules have to be viewed as parts of complex networks that change with time and location. Neural crest cells have to integrate these signals to ensure the generation of appropriate numbers of differentiating progeny. It will be important to determine how such signaling networks are established and how they elicit multiple signaling responses in neural crest cells to activate appropriate genetic programs. PMID- 17076284 TI - Neural crest stem cells. AB - Stem cells are defined by their ability to both self-renew and give rise to multiple lineages in vivo and/or in vitro. As discussed in other chapters in this volume, the embryonic neural crest is a multipotent tissue that gives rise to a plethora of differentiated cell types in the adult organism and is unique to vertebrate embryos. From the point of view of stem cell biology, the neural crest is an ideal source for multipotent adult stem cells. Significant advances have been made in the past few years isolating neural crest stem cell lines that can be maintained in vitro and can give rise to many neural crest derivatives either in vitro or when placed back into the context of an embryo. The initial work identifying these stem cells was carried out with premigratory neural crest from the embryonic neural tube. Later, neural crest stem cells were isolated from postmigratory neural crest, presumably more restricted in developmental potential. More recently it has been demonstrated that neural crest stem cell progenitors persist in the adult in at least two differentiated tissues, the enteric nervous system of the gut and the whisker follicles of the facial skin. In all cases, the properties of the stem cells derived reflect their tissue of origin and the potential of the progenitors becomes more restricted with age. In this chapter we will review this work and speculate on future possibilities with respect to combining our knowledge of neural crest gene function in the embryo and the manipulation of adult neural crest stem cells in vitro and eventually in vivo. PMID- 17076285 TI - Molecular bases of human neurocristopathies. PMID- 17076287 TI - Defining public health nursing. PMID- 17076286 TI - Evolution of the neural crest. AB - The recent advances in studies of the neural crest in vertebrates and the analysis of basal chordates using molecular and embryological approaches have demonstrated that at least part of the genetic programs and the cellular behavior were in place in nonvertebrate chordates before the neural crest evolved. Nevertheless, both the missing aspects and the close similarities found could explain why basal chordates lack a bona fide neural crest population, even though some migratory neurons and pigment cells have been recently identified in ascidians and amphioxus. PMID- 17076288 TI - Empowering nurses to lobby for change. PMID- 17076289 TI - The dilemma of aged-care funding. PMID- 17076290 TI - Ongoing concern about photo essay. PMID- 17076291 TI - Lessons learnt from managing a complex tuberculosis case. PMID- 17076292 TI - Taking a nursing service into the bush. PMID- 17076294 TI - Housing programme empowers people to make changes. PMID- 17076293 TI - From cradle to grave nursing. PMID- 17076295 TI - Meeting the health literacy needs of clients. PMID- 17076296 TI - Meeting the complex health needs of refugees. PMID- 17076297 TI - Releasing the bottleneck of mandated clinical hours. PMID- 17076298 TI - Coming to grips with nursing education funding. PMID- 17076299 TI - Ensuring history does not repeat itself. PMID- 17076300 TI - Planning for an influenza pandemic. PMID- 17076301 TI - [The impact of mood alterations on creativity]. AB - Basic elements of artistic (and other) creativity are the technical-professional skill and knowledge, the special talent and ability and the willingness or motivation; one of which being absent results in partially realised creativity like juvenile, frustrated or abandoned types, respectively. Psychometric scales have been developed to measure everyday and eminent creativity, which show that creativity correlates with higher psychoticism, impulsivity and venturesomeness scores and with lower neuroticism and conformity scores of the personality test employed in a general population. Among the psychological components of creativity are the cognitive processes, mood, motivation, and personality traits. Regarding mood, a theory of "inverted U" has been proposed as elevation of mood facilitates creativity to a certain point after what extreme increase has an adverse effect on achievement. Analysing psychopathology and creativity among various professions, higher rates of psychopathology, especially affective symptoms, have been found in art-related professions. Examples of immortal poets, writers, painters, sculptors and composers, having created invaluable cultural treasures for mankind, illustrate that many of them showed signs of mood alterations (unipolar or bipolar affective disorder spectrum) which were expressed in their artistic products. PMID- 17076302 TI - [Parallel processing of visual information]. AB - This is a survey on the function of parallel visual pathway with a special emphasis on its clinical implications. It is based on data in the literature and own results of our group. The paper primarily deals with the X, Y, W pathways and by the magnocellular, parvocellular and koniocellular visual pathways characterized by cells of various size as well as by nerve fibers of various thickness. Electrophysiological, microelectrode recording of single-unit activity makes the distinction between the pathways available in animal model. Much more difficulties arise if we intend to characterize the pathways in humans or to detect the selective damage of one of these pathways in patients. The non invasive diagnostic methods that could be used in the diagnosis are detailed here, too. Finally, the neurological, ophthalmological and psychological diseases are discussed in which a selective damage of any visual pathway is suspected. Summing it up, the survey provides evidences for the introduction of the novel concept of parallel pathways into the diagnostic aspects of ophthalmology, neurology and psychiatry. PMID- 17076303 TI - [The role of micro-arousals in the regulation of sleep]. AB - This work give a short account about a three decades research of the sleep microstructure. The studies, executed by the Strassbourg, Budapest and Parma schools, paved the way of exploring the participation of micro-arousals in the sleep regulation. It was shown that micro-arousals, not leading to instant arousal but influencing the later course of sleep are weaved into the network of sleep. A certain class of micro-arousals differs from the traditional desyncronisation-type and in a paradox way result a rebound like mobilisation of sleep-like activity with deltas and K-complexes. The desynchronisation- and synchronisation-type micro-arousals show different distribution along the sleep cyclicity and may play different role in sleep regulation. On the basis of the studies dealing with micro-arousals we can assume that beside the traditional long time constant, brain stem driven tonic chemical regulation, an other phasic regulation, with shorter time constant, associated with the micro-arousals, also exists. This kind of phasic regulation makes sleep flexible and possible to adapt the actual sleep course to the inner and outer demands. An other important role of micro-arousals in pathological sleeps is to provide a gate for the different pathological events, pinpointing the key-points where these events could be expected in the sleep process. PMID- 17076304 TI - The augmentation of clozapine treatment with electroconvulsive therapy. AB - OBJECTIVES: The assessment, in terms of safety and efficacy, of augmenting clozapine monotherapy, as well as combined psychopharmacotherapy involving clozapine, with electroconvulsive therapy (ECT). METHOD: Reviewed were the charts of patients who received clozapine-ECT treatment in the Department of Psychiatry and Psychotherapy of Semmelweis University between November 1999 and December 2003. RESULTS: During the studied period there were altogether 43 patients treated with the combination of clozapine and electroconvulsive therapy. In the schizoaffective group, the values for post-electroconvulsive therapy CGI were significantly lower than either in the catatonic (Z=-3.72, p<0.01) or in the hebephrenic (Z=-3.17, p<0.01) group. Among the groups created on the basis of the number of augmentation strategies utilized, the clozapine+3 group consisted of patients significantly older than the clozapine+ 1 group (Z=2.45, p=0.01). In the clozapine monotherapy group, the values for post-electroconvulsive therapy CGI were significantly lower than in any of the groups that had received a combination of augmentations (monotherapy-1 augmentation: Z=-3.01, p<0.01; monotherapy-2 augmentation: Z=-2.89, p<0.01; monotherapy-3 augmentation: Z=-2.41, p=0.01). CONCLUSIONS: According to our examinations, the augmentation of clozapine treatment with electroconvulsive therapy should be tried primarily on schizoaffective patients, in case the clozapine monotherapy is ineffective. The simultaneous use of different augmentation strategies is expected to increase only the side effects not the efficacy of the treatment. PMID- 17076305 TI - [Executive dysfunction in frontal lesions and frontal epilepsy]. AB - BACKGROUND: To explore the functions of the frontal lobe that are associated with high order cognitive and behavioral aspects such as the organization and execution of thoughts and behavior by neuropsychological methods is difficult. These so called executive functions are in close connection with the prefrontal thalamocortical circuits, damage of which can cause deficits in cognitive functions and even changes in personality. METHODS: The aim of this study is to present a neuropsychological battery for testing frontal lobe functions. 31 patients (with frontal epilepsy and/or frontal lesion) and 38 healthy control subjects participated in the study. The control subjects were matched to the patient group in age, gender and education. RESULTS: Comparing to the controls the patient group showed significant deficits in most of the measured executive functions, except two tests which show that the short time selective attention is preserved. We divided the patient group into three subgroups (frontal epilepsy only FLE, frontal lesion only FL, frontal lesion and epilepsy FLE+FL) and we found that except working memory deficits and problems in inhibition, there were no difference between the FLE patients and the control group. We found most frequently perseveration and errors in a strategy making task among the FL (mainly medial) patients. We didn't find any difference in these tests according to the lateralization of the lesion. CONCLUSION: In conclusion we found that working memory deficits and problems in inhibition differentiated the frontal patient group from the controls in all cases. We emphasize that in frontal epilepsy (with no reported MR lesion) there are the same type, however more limited neuropsychological alterations as in lesional frontal dysfunction. PMID- 17076306 TI - [Application of vertebroplasty, neuronavigation and kyphoplasty in the treatment of multiplex osteoporotic vertebral fractures--case report]. AB - Vertebroplasty is a image-guided therapeutic procedure, consisting of an injection of acrylic cement through a bone biopsy needle into a vertebral body. Main indication for vertebroplasty is painful vertebral body compression fracture due to osteoporosis. The procedure is an efficient mean with high success in pain release and prevention of further collapse of the treated vertebrae; however, the technique does not allow to realign the spine. Kyphoplasty was designed to address the kyphotic deformity. It involves the percutaneous placement of an inflatable bone tamp into a vertebral body (VB). Restoration of VB height and kyphosis correction is achieved by inflation of the tamp with contrast material liquid. After deflation a cavity is created that eases the cement application. The most modern way of guidance in spinal surgery is neuronavigation--the use of frameless stereotaxy. The system reformats patient-specific CT images acquired prior surgery, performs image fusion with intraoperative plain X-ray. Before the operation, the surgeon may create surgical plan and simulate advancement of a virtual instrument along one or more surgical trajectories. During surgery, the system tracks the position of specialized surgical instruments. All three modalities mentioned above have been applied in the treatment of our patient suffered from multiple osteoporotic vertebral body compression fractures. Using kyphoplasty an almost total VB height restoration could be achieved. The pain relief was more than 50% after both operation. PMID- 17076307 TI - [Aids-related toxoplasma-encephalitis presenting with acute psychotic episode]. AB - The most frequent neurological manifestations of the Acquired Immunodeficiency Syndrome-(AIDS) are Cerebral Toxoplasmosis, Primary Central Nervous System Lymphoma (PCNSL), Progressive Multifocal Leukoencephalopathy (PML) and AIDS encephalitis (AIDS-dementia complex, multinucleated giant cell encephalitis, HIV encephalopathy). Neurological complications usually occur in the advanced stages of the disease, and they are uncommon in the beginning as presenting illness, but may result in life-threatening condition or in death. Rarely the disease presents as a neuropsychiatric illness in an undiagnosed AIDS patient, delaying a proper diagnosis. We present the case of a 34 years old patient treated for AIDS-related Toxoplasma-encephalitis in our department. His illness started as an acute psychosis followed by rapid mental and somatic decline, leading to death in three months. His HIV-seropositivity was not known at his admission, and the extraneural manifestations were slight. The diagnosis was established by serology, imaging methods and histopathological investigation. After presenting the medical history and results of autopsy studies of the patient we discuss the problems of the differential diagnosis, especially regarding the findings of the imaging methods. PMID- 17076308 TI - [Novel forms of eating disorders]. PMID- 17076309 TI - Effect of spherical silica filler addition on immediate interfacial gap-formation in Class V cavity and mechanical properties of resin-modified glass-ionomer cement. AB - The aim of this study was to investigate how the addition of silanized spherical silica filler (SF) would influence the formation of summed, immediate interfacial gaps in Class V tooth cavities. Resin-modified glass-ionomer cement (RMGIC) is usually used for Class V restorations. As such, the following aspects of RMGIC were examined in correlation with summed interfacial gaps in the tooth cavity: setting shrinkage of cement in the Teflon mold, as well as mechanical properties in terms of compressive strength, diametral tensile strength, and flexural strength. Spherical silica filler was added to the RMGIC powder (Fuji II LC EM). For comparison purpose, untreated spherical silica filler (UF) was added too. When compared with the control (i.e., original RMGIC mixed with manufacturer recommended powder/liquid ratio), the addition of SF significantly decreased the formation of summed interfacial gaps in Class V cavities in the immediate condition. In particular, addition of 10 wt% SF increased the compressive strength by 56%, while diametral tensile strength was increased by 28% and flexural strength by 26%. PMID- 17076310 TI - In vitro pH analysis of active and arrested dentinal caries in extracted human teeth using a micro pH sensor. AB - The aim of this study was to evaluate the pH at the surface of active or arrested carious dentin using a micro pH sensor, and to compare the relationship between different pH measurement techniques. Twenty extracted carious teeth were divided into two groups, active or arrested caries, according to predefined clinical criteria before extraction. The surface pH values of carious dentin were measured using three methods: surface pH directly measured using a micro pH sensor (Direct); sectioned teeth measured using a pH-imaging microscope (Imaging) or micro pH sensor (Slice). For all techniques, statistically significant differences in pH values were observed between active and arrested dentinal caries (p<0.05). In addition, positive relations between the three pH measurement methods were found. In conclusion, Direct pH measurement using micro pH sensor might assist in caries lesion assessment and clinical treatment based on the concept of Minimal Intervention. PMID- 17076311 TI - Development of bonding system for resin core construction by MomegaA adhesive. AB - We designed self-etching primers consisting of a series of four N-methacryloyl omega-amino acids, MomegaA, of different methylene chain numbers for resin core construction. The interacted amount of MomegaA's carboxylic acid with root or crown dentin apatite was determined, and its effects on dentin bond durability examined. The addition of both dentin particles to the MomegaA solutions caused the carbonyl carbon peak of carboxylic acid in MomegaA to shift to a lower field, chiefly because of an acid-base interaction between carboxylic acid and calcium. Then, as the pKa value of MomegaA's carboxylic acid increased, the amount of carboxylic acid that interacted with calcium decreased. In terms of dentin bonding durability, the four tested MomegaA adhesives provided noticeably higher bond strengths of resin to root or crown dentin than ED Primer II. Therefore, from the perspective of restoring pulpless teeth with minimal intervention, carboxylated MomegaA adhesives seemed to be very useful as functional monomers for self-etching primers. PMID- 17076312 TI - Development of dental composite resin utilizing low-shrinking and low-viscous monomers. AB - To lower the viscosity of composite resins, experimental composite resins were produced using low-viscosity monomer mixtures of newly developed polyfunctional acrylates, and the mechanical and physical properties of the hardened composites were investigated. Mechanical (i.e., compressive, diametral tensile, and bending) strength of a polymer obtained from one new monomer mixture without fillers was similar to that of a bis-GMA/TEGDMA (2/1 weight ratio) based polymer. As for the hardened composites, the mechanical strength of composites produced using the new monomer mixtures showed a different tendency from that of bis-GMA based composites. Further, even the viscosity of composite pastes with high filler content was markedly lower than that of bis-GMA based composites. In terms of setting shrinkage, the composites consisting of new monomer mixtures exhibited significantly smaller shrinkage than the bis-GMA based composites, and decreased with increase in filler content. PMID- 17076313 TI - Silorane-based dental composite: behavior and abilities. AB - The purpose of this study was to examine the characteristics of an innovative composite material for dental restorations based on silorane--a monomer with a new chemical composition, and thereby compare the examined characteristics against those of well-known methacrylate-based composites. Degree of conversion at 2-mm and 6-mm depths as well as hardness, modulus of elasticity, and creep resistance through the middle of 6-mm high samples were measured. It was observed that up to 20 minutes after curing, curing time--and not irradiance--played the determinant role for a high degree of cure. No differences were registered between the two categories of material in terms of hardness. However, modulus of elasticity of the silorane-based material was slightly lower and the creep resistance higher than a methacrylate composite (Tetric EvoCeram). In conclusion, siloranes exhibited good mechanical properties comparable to those of clinically successful methacrylate-based composite materials. PMID- 17076314 TI - Chemical, morphological and thermal effects of 10.6-microm CO2 laser on the inhibition of enamel demineralization. AB - Studies have shown that enamel can be modified by pulsed CO2 laser to form a more acid-resistant substrate. This study evaluated the effects of a 10.6-microm CO2 laser on enamel surface morphology and chemical composition as well as monitored intrapulpal temperature changes during irradiation. Human teeth were irradiated with fluences of 1.5-11.5 J/cm2, and pulpal thermal as well as chemical and morphological modifications on enamel were assessed. The teeth were submitted to a pH-cycling model, and the mineral loss was determined by means of cross sectional microhardness. For all irradiated groups, intrapulpal temperature changes were below 3 degrees C. FT-Raman spectroscopy and scanning electron microscopy indicated that fluences as low as 6.0 J/cm2 were sufficient to induce chemical and morphological changes in enamel. Then, for fluences reaching or exceeding 10.0 J/cm2, laser-induced inhibitory effects on demineralization were observed. It was thus concluded that laser energy density in the range of 10.0 and 11.5 J/cm2 could be applied to dental enamel in order to produce chemical and morphological changes and reduce the acid reactivity of enamel without compromising the pulp vitality. PMID- 17076315 TI - Effect of conditioners on bond durability of resin composite to Nd:YAP laser irradiated dentin. AB - The aim of this study was to evaluate the effect of conditioners (sodium hypochlorite (NaOCl), Roth's ethylenediaminetetraacetic acid (EDTA), and phosphoric acid) on shear bond strength and morphology of Nd:YAP laser-irradiated dentin. In particular, the key focus was on the bond durability between resin composite and treated dentin after being subjected to thermocycling in artificial saliva between 5 degrees C and 55 degrees C. Results indicated that the application of phosphoric acid to laser-irradiated dentin produced a bond strength comparable to those using NaOCl and EDTA. Further, dentinal tubules which were closed after laser irradiation opened following the treatment with conditioners. When subjected to 3,000 thermocycles, the mean shear strength of the samples treated by the three conditioners following laser irradiation ranged from 9.7 to 12.6 MPa with a reduction of 25-33%--a reduction rate lower than that obtained using acid etching alone (50%). Among the three conditioners tested, only phosphoric acid treatment demonstrated an enhanced effect on bond durability of laser-irradiated dentin. PMID- 17076316 TI - Surface modification of titanium with hydrothermal treatment at high pressure. AB - Surface modification of titanium was investigated by means of hydrothermal treatment with a maximum pressure of 6.3 MPa (280 degrees C temperature) in CaO solution or water to improve bioactivity and biocompatibility. As a result, calcium titanate was formed on the titanium surface. Moreover, titanium oxide and titanium hydroxide layers on the surface increased as temperature and pressure increased. The surface-modified titanium was also immersed in a simulated body fluid (SBF) to estimate its bioactivity. Needle-like apatite precipitation was observed on all hydrothermal-treated titanium surfaces after immersion in SBF for four weeks. In particular, the apatite precipitation of titanium treated with 6.3 MPa in CaO solution was clearer and larger in amount than those of all other hydrothermal-treated specimens. Further, the amount of precipitate corresponded to the thickness of the surface-modified layer and the amount of calcium in the surface layer. The results suggested that surface modification of titanium with high-pressure hydrothermal treatment seemed to improve bioactivity and biocompatibility. PMID- 17076317 TI - Mode superposition transient dynamic analysis for dental implants with stress absorbing elements: a finite element analysis. AB - The purpose of this study was to analyze the dynamic behavior of a dental implant with a stress-absorbing element, using dynamic analysis. Two model types, stress absorbing model with a resilient stress absorber made of polyoxymethylene and non stress-absorbing model with rigid titanium, were employed. In both model types, the implant was 4.0 mm in diameter and 13.0 mm in length and placed in the mandibular first molar region. Shapes of the finite element implant and implant bone were modeled using computer-aided design. All calculations for the dynamic analysis were performed using the finite element method. It was found that the stress-absorbing model had a lower natural frequency than the non-stress absorbing model. In addition, the stress-absorbing model had a higher damping effect than the non-stress-absorbing model. It was concluded that mode superposition transient dynamic analysis is a useful technique for determining dynamic behavior around dental implants. PMID- 17076318 TI - Ultrastructural characterization of tissue response to sintered carbonate apatite in rabbit bone. AB - Our previous in vivo and in vitro studies revealed excellent tissue biocompatibility and osteoconductivity of porous sintered carbonate apatite (CA). The present study focused on the ultrastructural details of cells involved in the degradation of CA and new bone formation. Electron microscopy indicated that multinucleated giant cells (MNGCs) were actively involved in CA resorption. MNGCs extended their irregular cytoplasmic protrusions deeply into the interstitial spaces between CA particles. Endophagosomes were formed by encircling partially dissolved or intact CA crystals via the development of pseudopodia-like cytoplasmic protrusions, the configuration of which was somewhat different from that of the typical ruffled border of bone-resorbing osteoclasts. Subsequently, most CA particles in MNGCs were irregular in shape, suggesting that acidic degradation of CA occurred mainly intracellularly. Mononuclear cells, such as macrophage-like and/or fibroblast-like cells, also took up and degraded some CA. Growth of very thin needle-like crystals was observed in close association with CA. Osteoblasts directly faced the CA and secreted osteoid matrix. At the CA-bone interface, an electron-dense and homogeneous thin layer free of collagen fibers was sometimes observed, suggesting an involvement in CA-bone bonding. PMID- 17076319 TI - Shear bond strengths of different substrates bonded to lithium disilicate ceramics. AB - The aim of this study was to evaluate the shear bond strengths of pre- and untreated resin core materials to lithium disilicate ceramics. Bond strengths to dental hard tissues served as controls. Ceramic cubes (IPS-Empress II) were luted either with a one-step (Variolink II/Excite DSC) or multiple-step total etching bonding system (Tetric Flow/Syntac Classic) to ground surfaces of human enamel, dentin, and resin core materials (Clearfil Core, Multicore). Resin core materials were additionally pretreated with hydrofluoric acid (HF) or were silica-coated (CoJet System). Shear bond strengths were determined after 24-hour water storage (n=10) and thermocycling (TC) (n=10; 2000 cycles, 5-55 degrees C, 30 seconds). Bond strengths to enamel, dentin, and silica-coated composites were significantly higher compared to untreated and HF-pretreated composites (p<0.05; Tukey B). Indeed, silica coating of the composite resins significantly increased the bond strength to ceramics (p<0.05; Tukey B). Due to the lower bond strength values of ceramics to untreated composite resins (as compared to enamel and dentin), any indication for a resin core build-up prior to the preparation of a ceramic restoration should be considered carefully. PMID- 17076320 TI - Effect of incremental filling technique on adhesion of light-cured resin composite to cavity floor. AB - The purpose of this study was to evaluate the effect of various incremental filling techniques on adhesion between composite and cavity floor using light cured resin composite. Black ABS resin and hybrid resin composite were used as mold materials--instead of dentin--for the preparation of cavities, and standardized to 5x5x5 mm. Each cavity was then treated with a bonding system (Clearfil SE bond). Resin composite (Clearfil Photo Core) was placed on the bonding resin using different incremental filling techniques or in bulk and irradiated for a total of 80 seconds using a halogen light unit. Specimens were subjected to the micro-tensile bond test at a crosshead speed of 1 mm/min. Data were analyzed by two-way ANOVA. The results indicated that an incremental filling technique was more effective in improving adhesion to the cavity floor than a bulk filling technique. PMID- 17076321 TI - Evaluation of a new fluoride-releasing one-step adhesive. AB - In this study, a newly developed S-PRG (surface pre-reacted glass-ionomer) filler containing one-step adhesive, called SI (SI-IB551, Prototype), was evaluated regarding its fluoride release behavior. As a result, fluoride ion distribution in the tooth structures and acid resistance of cavity margins were also evaluated. In addition, Absolute (a fluoride-releasing one-step adhesive; AB) was evaluated in comparison to G-Bond (a non-fluoride-releasing one-step adhesive; GB) which was used as a negative control. Concentration of fluoride released was measured using a fluoride ion selective electrode after immersion of each material in distilled water. Fluoride distribution in tooth-adhesive interfacial areas was examined with EPMA following cavity preparation in human premolars and having treated resin restorations with each material. To evaluate acid resistance, restored specimens were immersed in an acetic acid buffer (0.2 M, pH 4.5) for 12 hours and then the cavity margins were observed using scanning electron microscopy. Amounts of fluoride released from AB and SI were significantly greater than that from GB. Further, significant differences in fluoride release were detected between AB and SI. A layer of increased fluoride density was clearly detected at the enamel- and dentin-SI interfaces. In terms of acid resistance, an acid resistance zone was also formed adjacent to the tooth adhesive interface of AB and SI specimens. However, in GB specimen, acid resistance zone was not observed. These findings suggested that one-step adhesives displayed a favorable fluoride release property, thereby contributing positively to inhibition of recurrent caries. PMID- 17076322 TI - Electrochemical behavior of cast Ti-Ag alloys. AB - Anodic polarization tests were performed in 0.9% NaCl and 1% lactic acid solutions to characterize the relationship between the corrosion behavior and the microstructures of cast Ti-Ag (5-40% Ag) alloys. The anodic polarization curves for the Ti-Ag alloys up to 17.5% Ag were similar to those for pure titanium in both solutions. On the other hand, an abrupt increase in the current density was observed for the alloys with more than 20% Ag in the NaCl solution and with more than 27.5% Ag in the lactic acid solution. The microstructures of the corroded alloy surfaces indicated the deterioration of precipitated intermetallic compounds along the grain boundaries. The Ti-Ag alloys up to 17.5% Ag had excellent corrosion resistance similar to that of pure titanium. The alloys with 20-25% Ag may be also used as dental alloys, since they passivated again immediately after preferential dissolution in the NaCl solution. PMID- 17076323 TI - Modification of 4-META/MMA-TBB resin for safe debonding of orthodontic brackets- influence of the addition of degradable additives or fluoride compound. AB - The purpose of this study was to evaluate the performance of modified 4-META/MMA TBB resin cements (Superbond C&B) in terms of debonding orthodontic brackets easily and safely from enamel without the loss of proper bracket bond strength. Poly (DL-lactide-co-glycolide) (PLGA), calcium fluoride (CaF2), or alpha tricalcium phosphate (alpha-TCP) was added to the polymer powder of 4-META/MMA TBB resin, and the shear bond strength of orthodontic brackets to human enamel using modified resins was measured before and after 10,000-cycle thermal cycling test between 5 degrees C and 55 degrees C. The modified resins tended to provide lower bond strength compared with the original 4-META/MMA-TBB resin. However, alpha-TCP- or CaF2-modified resin showed no significant differences in bond strength before and after thermal cycling. Moreover, there was a tendency of more residual resin remaining on the tooth surface after debonding, thereby suggesting a lower risk of enamel fracture. In conclusion, alpha-TCP- or CaF2-modified 4 META/MMA-TBB resin seemed to allow easy and safe debonding of orthodontic brackets without loss of proper bracket bond strength. PMID- 17076324 TI - Estrogenic activity of phthalate esters by in vitro VTG assay using primary cultured Xenopus hepatocytes. AB - Estrogenic activity of phthalate esters in dental soft resins was evaluated with an amphibian system consisting of a vitellogenin (VTG)-detecting Enzyme-Linked Immunosorbent Assay and a primary-cultured hepatocyte assay using adult male Xenopus laevis. In particular, phthalate esters--Di-n-butyl phthalate (DBP), Butyl phthalyl butyl glycolate (BPBG), Benzyl butyl phthalate (BBP), and Benzyl benzoate (BB)--were investigated. Bisphenol A (BPA) was prepared for comparison with these chemicals, and 17beta-estradiol (E2) was used as a positive control. The chemicals were diluted in dimethyl sulfoxide (DMSO) to obtain final concentrations ranging from 10(-11) to 10(-4) mol/l. BPA induced estrogenic activity at a concentration of 1.1x10(-6) mol/l, while E2 showed at 4.1x10(-11) mol/l. DBP, BBP, BB, and BPBG showed no estrogenic activity at concentrations between 4x10(-7) mol/l and 1x10(-4) mol/l. The latter result indicated that these phthalate esters might be metabolically transformed into non-estrogenic substances in Xenopus hepatocytes. Furthermore, this study demonstrated that through in vitro metabolism assessment, the estrogenic activity of chemical substances could be directly detected in terms of VTG secretion in primary cultured Xenopus hepatocytes. PMID- 17076325 TI - Influence of elasticity on gap formation in a lining technique with flowable composite. AB - The purpose of this study was to investigate the effect of flowable composites as liners for direct composite restorations, with key focus on the elastic moduli of flowable and condensable composites. After treating the composite mold cavity surface with an adhesive system, one of the flowable composites was placed as a 1 mm-thick layer on the cavity floor and irradiated for 20 seconds. The rest of cavity was subsequently filled with a condensable composite and irradiated for 40 seconds. Gap formation at both interfaces--between the cavity floor and flowable composite, and between the flowable and condensable composites--was examined. No gaps were detected at the interface between the cavity floor and flowable composite. Gap percentage at the interface between the flowable and condensable composites was dependent on the difference in elastic modulus. It was concluded that flowable composite with high elastic modulus could inhibit gap formation between flowable and condensable composites. PMID- 17076326 TI - Evaluation of antibacterial and fluoride-releasing adhesive system on dentin- microtensile bond strength and acid-base challenge. AB - This study evaluated the influence of a fluoride-containing adhesive on microtensile bond strength (microTBS) to dentin, as well as analyzed the dentin adhesive interface after acid-base challenge. Experimental groups were: G1- Clearfil SE Bond control (SE); G2--Clearfil Protect Bond control (PB); G3- Primer[SE]/Adhesive[PB]; G4--Primer[PB]/Adhesive[SE]. For microTBS evaluation, dentin surfaces were ground, bonded, and composite resin crowns were built up to obtain beams to be tested. For interfacial analysis, adhesive system was applied on dentin surface and a low-viscosity resin was placed between two dentin disks. Then, the specimens were subjected to acid-base challenge, sectioned, and polished to be observed by SEM. microTBS data showed no statistical differences among the groups (GI: 51.3, G2: 47.6, G3: 55.0, G4: 53.9; mean in MPa). Through SEM, it was observed that a thick acid-base resistant zone adjacent to the hybrid layer was created only when the fluoride-releasing adhesive was used. In conclusion, the presence of fluoride in an adhesive contributed significantly to preventing secondary caries, and did not interfere with dentin-adhesive bond strength. PMID- 17076327 TI - Experimental binder-free investments reused to cast dental precious alloys. AB - This study aimed to develop reusable dental investments. SiO2 and MgO were selected as refractory materials to prepare three types of investment (coded as 60S-40M, 80S-20M, 100S) with 40, 20, and 0 wt% of MgO. Each type of investment was reused twice. Thermal expansion and compressive strength were examined and statistically evaluated by ANOVA. To evaluate fit of castings, full crowns were cast by using a commercial Au-Ag-Pd alloy with all investment types. Marginal fit was statistically analyzed by cement thickness. It was found that although MgO strengthened the mold, it had little influence on expansion. The strength of 60S 40M was the highest, and 100S had the greatest advantage with regard to thermal expansion. In the evaluation for clinical applicability, all investments were able to cast successfully, but their castings might be undersized. Among the experimental binder-free investments reused for dental casting in this study, 100S in particular showed to be a good candidate for repeated fabrication of precision fit castings. PMID- 17076328 TI - Surface modification of poly (L-lactide) by atmospheric pressure plasma treatment and cell response. AB - This study investigated the influence of atmospheric pressure plasma treatment on the surface properties and cell response of poly(L-lactide) (PLLA) samples. The samples were analyzed by means of X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), micro- and nanosurface roughness, water contact angle, and zeta potential. Furthermore, cell adhesion assay and cell proliferation assay on the samples were carried out using MC3T3-E1 cells. Plasma treatment significantly increased the oxygen content of the samples and decreased the contact angle and zeta potential of the samples, resulting in hydrophilic surfaces. Further, plasma treatment of the samples also enhanced the number and growth of adhering MC3T3-E1 cells. These results therefore indicate that plasma treatment is effective for surface modification and cell responses. PMID- 17076329 TI - Design of new phosphonic acid monomers for dental adhesives--synthesis of (meth) acryloxyalkyl 3-phosphonopropionates and evaluation of their adhesion-promoting functions. AB - The aims of this study were to synthesize new phosphonic acid monomers, and to examine their bonding performance. Four kinds of newly designed phosphonic acid monomers were synthesized, and seven experimental composite-type adhesive resins comprising the synthesized monomers (3 wt%), with BPO/DEPT/BPBA initiator were prepared. Tensile bond strengths of the adhesive resins to enamel and metal were measured with a universal testing machine at a crosshead speed of 1.0 mm/ min. The synthesized adhesive monomers were light yellow viscous liquids with 32.5% 49.3% yields, and identified by 1H NMR, IR and elemental analysis to be (meth)acryloxyalkyl 3-phosphonopropionates [R-P(=O) (OH)2] (5-MPPP, 6-MHPP, 6 AHPP, 10-MDPP). It was found that the newly developed phosphonic acid monomers with BPO/DEPT/BPBA initiator attained strong adhesion to both unetched, ground enamel and sandblasted Ni-Cr alloy with good durability. They exhibited significantly higher bond strengths than conventional phosphorous-containing monomers such as MEPP and VBPA (p <0.01). These findings indicate that the experimental phosphonic acid monomer-containing adhesive resins have potential prosthodontic and orthodontic applications, especially as self-etching, non rinsing orthodontic adhesive resins. PMID- 17076330 TI - Contraction gap versus shear bond strength of dentin adhesive in sound and sclerotic dentins. AB - To evaluate the effect of a dentin adhesive on sclerotic dentin, contraction gap width and shear bond strength were measured. Dentin cavity wall was pretreated with an experimental dentin bonding system with and without a dentin primer, or with a commercial dentin bonding system. In the experimental dentin bonding groups, contraction gap width of sclerotic dentin was significantly smaller than that of sound dentin when the cavity was not primed with glyceryl monomethacrylate. For each individual tooth, the correlation between contraction gap width and shear bond strength was insignificant. In conclusion, the bonding efficacy of dentin bonding systems to sclerotic dentin was superior to that of sound dentin. Further, it was determined that it was impossible to detect the interaction between the polymerization contraction stress of resin composites and the efficacy of dentin adhesives by measuring bond strength. PMID- 17076331 TI - Relationship between colors of carious dentin and laser fluorescence evaluations in caries diagnosis. AB - This in vitro study investigated the relationship between assessments of dentin caries using a laser fluorescence device (DIAGNOdent) and a caries detector dye during caries removal. The dentin of eight extracted carious molars was removed at 300-microm interval points from the dentin surface toward the pulp chamber. Before and after each removal, images of the carious surfaces were taken in association with color-matching stickers (for color correction) and the surfaces were evaluated by DIAGNOdent based on fluorescence emission from the tooth surface. For the L* values (CIE 1976 L*a*b* color system), there was a strong negative correlation between DIAGNOdent results and the corrected L* values of the carious surfaces (Pearson's correlation coefficient: -0.853); additionally, there was a significant correlation between them (p<0.05). However, there were no significant correlations between the DIAGNOdent results and the corrected a* and b* values of the carious surfaces (Pearson's correlation coefficients: 0.108 and 0.018 respectively). In conclusion, DIAGNOdent was shown to be applicable for caries diagnosis during caries removal. PMID- 17076332 TI - Effects of apatite foam combined with platelet-rich plasma on regeneration of bone defects. AB - The objective of this study was to investigate the regenerative effects of apatite foam (AF) combined with platelet-rich plasma (PRP) on bone defects. Critical-sized defects in the tibia of rats were filled with randomly distributed combinations of AF with and without PRP. The animals were killed after three, six, and 12 weeks, and their tissue responses were histologically examined. At three weeks, we found no significant differences in bone regeneration against control group (21.9 +/- 3.1%) when PRP (20.3 +/- 4.2%) and AF (21.6 +/- 2.9%) were used independently of each other. In contrast, significantly (p<0.01) larger amount of bone (32.3 +/- 6.5%) was formed when the defect was filled with PRP incorporated AF. At six weeks, both PRP (38.1 +/- 3.2%) and AF (39.6 +/- 7.8%) showed significantly (p<0.05) higher rates of bone regeneration than the control, even though they were used independently. Moreover, the amount of regenerated bone significantly (p<0.01) increased in the defect filled with PRP-incorporated AF (76.1 +/- 8.2%). We concluded, therefore, that the combination of PRP and AF may be useful for the regeneration of defected bone. PMID- 17076333 TI - Detailed consideration of physicochemical properties of CO3apatites as biomaterials in relation to carbonate content using ICP, X-ray diffraction, FT IR, SEM, and HR-TEM. AB - CO3apatites with different carbonate contents were synthesized at 60 +/- 1 degrees C and pH 7.4 +/- 0.2 under different carbonate concentrations (0-0.3 mol/L) in the supplied solutions. Their physicochemical properties were analyzed using various methods. Inductively coupled plasma gave accurate chemical analysis data for calcium and phosphate contents. X-ray diffraction analysis showed a clear chemical shift at high carbonate content. A CO3(2-) absorption peak area approximately proportional to carbonate content was observed through Fourier transmission infrared spectroscopy. Scanning electron microscopy and high resolution transmission electron microscopy revealed a dramatic change of the crystal shape. Osteoblast proliferation at the surface of each CO3apatite collagen sponge indicated that osteoblasts deformed to expand and cover the surface of the sponge, and appeared to adhere well to the sponge. PMID- 17076334 TI - Accuracy of temporary laser welding of FPDs by Nd:YAG laser in the oral cavity. AB - The purpose of this study was to investigate the accuracy of temporary fixation with laser welding for fixed partial dentures (FPDs). Five kinds of experimental FPD with different welding/soldering gaps were fabricated (0, 20, 50 microm for welding; 300 microm for soldering). Then, FPDs were temporary-fixed by laser welding or with a self-curing resin. Fixation accuracy was evaluated by the change in distance and the angular deformation between two retainers. The change in distance and the angular deformation between two retainers of the FPD without welding/soldering gap were significantly larger than the other FPDs (p<0.05). With due consideration to the displacement of teeth or implants especially in the mesiodistal direction, and by taking into account the inevitable errors of the indirect method, it seemed reasonable to provide a welding space of approximately 20 microm. PMID- 17076335 TI - Retrospective study of five-year clinical performance of direct composite restorations using a self-etching primer adhesive system. AB - This study was conducted to assess the five-year clinical performance of composite restorations using a self-etching primer adhesive system, Clearfil Liner Bond II. Restorations were rated using the following scale: A--Good; B- Clinically acceptable; C--Clinically unacceptable; or D--Already replaced. The ratings were analyzed using Freedman's test (p=0.01) and Steel-Dwass test (p=0.05). A total of 83 restorations from 36 patients were identified from treatment records. Twenty-six restorations (31.3%) presented A rating and 36 restorations (43.4%) were clinically acceptable (B rating), while five (6.0%) received C rating and 16 (19.3%) had already been replaced (D rating). The mean survival rate of serviceable restorations (i.e., combined A and B ratings) after five years was 74.7%. It was found that cavity form, including cavity size, and occlusal contact seemed to influence longevity. PMID- 17076336 TI - Fabrication of biporous low-crystalline apatite based on mannitol dissolution from apatite cement. AB - Biporous (macro- and microporous) calcium phosphate gains much attention as a bone substitute material because of its large surface area and that it improves cell penetration. In the present study, we evaluated the feasibility of biporous, low-crystalline apatite based on dissolution of mannitol from self-setting apatite cement (Biopex). Mannitol--known as a biocompatible, easily dissolved monosaccharide alcohol--was recrystallized to obtain larger crystals. It was crushed with pestle and mortar, sieved to obtain crystals which passed through a 500-microm mesh but which remained against a 300-microm mesh, and then used as porogen. Although Biopex containing 60 wt% mannitol was not able to be taken out of the mold, addition of mannitol caused no initial setting inhibition to Biopex if the amount was 40 wt% or less. Similarly, transformation to apatitic product was confirmed when the apatite cement was immersed in 0.9% saline kept at 37 degrees C for seven days. The set mass became low-crystalline, biporous apatite with approximately 60% porosity. PMID- 17076337 TI - Shear bond strengths of four resin bonding systems to two silica-based machinable ceramic materials. AB - The purpose of this study was to evaluate the bond strength between four bonding systems (GC Ceramic Primer and Linkmax HV (CP+LM), Clapearl Bonding Agent and Clapearl DC (CBA+CL), Clearfil Mega Bond Porcelain Bonding kit and Panavia F2.0 (MB+PF), and RelyX Ceramic Primer and RelyX ARC (RC+RA)) and two machinable ceramics (Vitablocs Mark II, VMII and GN-I ceramic block, GNI). Shear bond strength was determined after 24-hour immersion in water or after thermocycling of 20,000 cycles. It was found that the post-thermocycling bond strength of each bonding system to VMII was significantly higher than that to GNI. VMII showed no significant differences between CP+LM, MB+PF, and CBA+CL--the values of which were higher than that of RC+RA. Regarding GNI, CP+LM showed the greatest bond strength after thermocycling among the four bonding systems used. It was concluded that the crystalline phase of the ceramics used might have an effect on bond strength. PMID- 17076338 TI - Effect of surface condition of dental zirconia ceramic (Denzir) on bonding. AB - Yttria partially stabilized zirconia (YPSZ) ceramics are suitable for dental and medical use because of their high fracture toughness and chemical durability. The purpose of this study was to examine the bonding behavior of a dental YPSZ ceramic, Denzir. After being subjected to various surface treatments, Denzir specimens were bonded to each other using an adhesive resin composite, glass ionomer, or zinc phosphate cement. Bonding strength was then determined by the shearing test. No significant differences (p>0.05) were observed between SiC- and Al2O3-blasted specimens. In all surface treatments, the shear bond strength significantly (p<0.05) increased in the order of adhesive resin composite cement > glass ionomer cement > zinc phosphate cement. Moreover, silanization with methacryloxy propyl trimethoxysilane slightly increased the bonding strength of the adhesive resin composite cement. PMID- 17076339 TI - Effects of welding pulse energy and fluoride ion on the cracking susceptibility and fatigue behavior of Nd:YAG laser-welded cast titanium joints. AB - In this study, the cracking susceptibility and fatigue behavior of Nd:YAG laser welded cast Ti joints (welding pulse energy: 11, 15, and 18 J) in fluoride containing (0 and 0.5% NaF) artificial saliva were evaluated using constant elongation rate test (CERT) and fatigue test (FT), respectively. Both CERT and FT were also carried out in open air as controls. Results showed that increasing the welding energy increased the elongation and fatigue life, but decreased the tensile strength, of cast Ti joints in open-air environment. With a welding energy of 11 J, the fluoride ions in the artificial saliva increased the cracking susceptibility and decreased the fatigue life of Ti joints. When the welding energy exceeded 15 J, the presence of fluoride ions still increased the cracking susceptibility, but did not reduce the fatigue life of Ti joints. Rupture of Ti joints--if it occurred--occurred only at the welded metal (versus the non-welded part). PMID- 17076340 TI - Figure-ground assignment in pigeons: evidence for a figural benefit. AB - Four pigeons discriminated whether a target spot appeared on a colored figural shape or on a differently colored background by first pecking the target and then reporting its location: on the figure or the background. We recorded three dependent variables: target detection time, choice response time, and choice accuracy. The birds were faster to detect the target, to report its location, and to learn the correct response on figure trials than on background trials. Later tests suggested that the pigeons might have attended to the figural region as a whole rather than using local properties in performing the figure-background discrimination. The location of the figural region did not affect figure-ground assignment. Finally, when 4 other pigeons had to detect and peck the target without making a choice report, no figural advantage emerged in target detection time, suggesting that the birds' attention may not have been automatically summoned to the figural region. PMID- 17076341 TI - Likelihood ratio, optimal decision rules, and correct response probabilities in a signal detection theoretic, equal-variance Gaussian model of the observer in the 4IAX paradigm. AB - This article provides a synthetic account of the likelihood ratio, optimal decision rules, and correct response probabilities in a signal detection theoretic model of the observer in the dual-pair comparison, or four-interval AX (4IAX), paradigm. The model assumes a static sampling process, resulting in four, equal-variance normally distributed (i.e., Gaussian) observations on each trial First, a likelihood ratio equation allowing for an arbitrary degree of correlation between observations is provided. Specific solutions for the cases of independent and highly correlated observations are then derived. It is shown that these solutions, and the associated decision rules, correspond to those provided independently in earlier publications. A modified 4IAX paradigm involving, as a standard, an additional stimulus (C) located medially between the A and the B stimuli is also considered. It is shown that the optimal (static, equal-variance, Gaussian) decision model for this paradigm is unaffected by correlation between observations and is equivalent to the standard 4IAX with highly correlated observations. Finally, we consider how, under the considered (static, equal variance, Gaussian) model, the proportion of correct responses in the different versions of the 4IAX paradigm is related to d', and a solution for the case of independent observations is provided. PMID- 17076342 TI - Cross-trial priming in visual search for singleton conjunction targets: role of repeated target and distractor features. AB - Kristjansson, Wang, and Nakayama (2002) demonstrated that visual search for conjunctively defined targets can be substantially expedited ("primed") when target and distractor features are repeated on consecutive trials. Two experiments were conducted to examine whether the search response time (RT) facilitation on target-present trials results from repetition of target-defining features, distractor features, or both. The experiments used a multiple conjunctive search paradigm (adapted from Kristjansson et al., 2002), in which the target and distractor features were varied (i.e., repeated) independently of each other across successive trials. The RT facilitation was numerically largest when both target and distractor features were repeated, but not significantly larger than that when only distractor features were repeated. This indicates that cross-trial priming effects in conjunctive visual search result mainly from the repetition of distractor, rather than target, features. PMID- 17076343 TI - Effects of the orientation of moving objects on the perception of streaming/bouncing motion displays. AB - In this study, we examined the contribution of the orientation of moving objects to perception of a streaming/bouncing motion display. In three experiments, participants reported which of the two types of motion, streaming or bouncing, they perceived. The following independent variables were used: orientation differences between Gabor micropatterns (Gabors) and their path of motion (all the experiments) and the presence/absence of a transient tone (Experiment 1), transient visual flash (Experiment 2), or concurrent secondary task (Experiment 3) at the coincidence of Gabors. The results showed that the events at coincidence generally biased responses toward the perception of bouncing. On the other hand, alignment of Gabors with their motion axes significantly reduced the frequency of bounce perception. The results also indicated that an object whose orientation was parallel to its motion path strengthened the spatiotemporal integration of local motion signals along a straight motion path, resulting in the perception of streaming. We suggest that the effect of collinearity between Gabors and their motion path is relatively free from the effect of attention distraction. PMID- 17076344 TI - More than meets the eye: the effect of planned fixations on scene representation. AB - Scene memory frequently includes a swath of unseen layout beyond a photograph's boundaries (boundary extension [BE]; Intraub and Richardson, 1989). Might it be affected by the viewer's plan to shift fixation near a view boundary? When photographs were centrally fixated (500 msec), BE occurred following a 2-sec masked interval (Experiment 1). In Experiments 2-4, a cue during the first fixation signaled viewers to fixate an object near the left or right boundary. The picture was masked before the eyes landed. BE occurred on the cued side and on the top and bottom, but not on the uncued side. This relatively accurate performance on the uncued side suggests that inhibition of a movement to one side (in a competitive task) may also inhibit extrapolation of layout. BE on the to-be fixated side, however, supports the idea that anticipatory representation of layout is an adaptive error that may aid the spatial integration of successive views. PMID- 17076345 TI - The effects of stimulus rotation and familiarity in visual search. AB - Rauschenberger and Yantis (2006) observed that an intersecting circle-line combination enjoyed significantly greater search efficiency when it was oriented to resemble a "Q" than when it was oriented so that the intersecting line was vertical (cf. Treisman and Souther, 1985). Although a control experiment made it unlikely that the obliqueness of the line was responsible for the observed benefit, there was no direct evidence that this benefit was attributable to the "Q-ness" of the stimulus. In the present study, a subset of Rauschenberger and Yantis's experiments was repeated with Chinese subjects, who had never been exposed to the Latin alphabet. For these subjects, there was no benefit for the"Q"-like stimulus, in contrast to the results of Rauschenberger and Yantis's study. These results show that a simple 45 degrees rotation of a stimulus can affect search efficiency significantly--but only when this rotation bestows meaning, or familiarity, to that stimulus. PMID- 17076346 TI - Visual search for a circular region perceived as a figure versus as a hole: evidence of the importance of part structure. AB - To study contour curvature polarity, we compared strictly convex regions (circular figures) with strictly concave regions (circular holes). We tested for an asymmetry between visual searches for concavities and those for convexities. We found that providing a preview of the background benefited search for concavities (holes) more than it did search for convexities (figures) and that for convex figures, nearer targets were responded to more quickly. Importantly, however, we failed to find any support for the hypothesis that concave targets are inherently more salient. We conclude that previous findings in the literature, which have been taken to indicate preferential processing of concavities, due to their increased salience, are more likely the result of an early computation of part structure based on concavities. PMID- 17076347 TI - Contextual cuing based on spatial arrangement of color. AB - Previous studies have demonstrated that, in visual search, repetition of a target's "context" significantly improves search performance (contextual cuing; Chun & Jiang, 1998). Context in those studies was defined as either spatial configuration or featural combination. In the present work, we demonstrate that repeating the spatial arrangement of object colors (conjunction of spatial configuration and featural combination) also leads to contextual cuing (Experiment 1). Experiments 2 and 3 rule out the possibility that the contextual cuing we find in Experiment 1 is caused only by the spatial configuration of color patches. Experiment 4 rules out the possibility that it is caused only by featural combination. Experiments 5 and 6 demonstrate that color-arrangement based contextual cuing is an unconscious process. Experiments 7 and 8 demonstrate that contextual cuing seems to be more effective onchromatic (hue) variation than on achromatic (luminance) variation. In sum, our results demonstrate that contextual cuing is not merely associated learning in separate domains; it is a more integrated process than has perhaps been appreciated. PMID- 17076348 TI - Does attention alter appearance? AB - Abrupt onsets in the visual field can change the appearance of subsequent stimuli, according to one interpretation, by engaging an attentional mechanism that increases effective stimulus contrast. However, abrupt onsets can also engage capacity-unlimited and thus attention-independent sensory mechanisms. We conducted a series of experiments to differentiate the sensory and attentional accounts. Observers compared the contrasts of uncued low-contrast peripheral targets with simultaneous targets cued by one of three cue types with different sensory attributes: white or black peripheral abrupt onsets and central gaze direction cues devoid of sensory activity near the target locations. Each cue facilitated the perception of perithreshold targets; however, the white abrupt onsets increased the perceived contrast of suprathreshold targets, whereas the black abrupt onsets tended to reduce the perceived contrast, and the gaze direction cues had no significant effect. The effectiveness of the gaze direction cues in automatically orienting attention was demonstrated in a control experiment in which they consistently speeded response times. The results suggest that sensory interaction, and not attention, is responsible for changes in appearance. PMID- 17076349 TI - Understanding the object benefit in visual short-term memory: the roles of feature proximity and connectedness. AB - Past research has identified visual objects as the units of information processing in visual short-term memory (VSTM) and has shown that two features from the same object can be remembered in VSTM as well (or almost as well) as one feature of that object and are much better remembered than the same two features from two spatially separated objects. It is not clear, however, what drives this object benefit in VSTM. Is it the shared spatial location (proximity), the connectedness among features of an object, or both? In six change detection experiments, both location/proximity and connectedness were found to be crucial in determining the magnitude of the object benefit in VSTM. Together, these results indicate that location/proximity and connectedness are essential elements in defining a coherent visual object representation in VSTM. PMID- 17076350 TI - Acuity of auditory images in pitch and time. AB - We examined the pitch and temporal acuity of auditory expectations/images formed under attentional-cuing and imagery task conditions, in order to address whether auditory expectations and auditory images are functionally equivalent. Across three experiments, we observed that pitch acuity was comparable between the two task conditions, whereas temporal acuity deteriorated in the imagery task. A fourth experiment indicated that the observed pitch acuity could not be attributed to implicit influences of the primed context alone. Across the experiments, image acuity in both pitch and time was better in listeners with more musical training. The results support a view that auditory images are multifaceted and that their acuity along any given dimension depends partially on the context in which they are formed. PMID- 17076351 TI - Hierarchical attention in discriminating objects at different levels of specificity. AB - An implicit assumption of studies in the attentional literature has been that global and local levels of attention are involved in object recognition. To investigate this assumption, a divided attention task was used in which hierarchical figures were presented to prime the subsequent discrimination of target objects at different levels of category identity (basic and subordinate). Target objects were identified among distractor objects that varied in their degree of visual similarity to the targets. Hierarchical figures were also presented at different sizes and as individual global and local elements in order to investigate whether attention-priming effects on object discrimination were due to grouping/parsing operations or spatial extent. The results showed that local processing primed subordinate object discriminations when the objects were visually similar. Global processing primed basic object discriminations, regardless of the similarity of the distractors, and subordinate object discriminations when the objects were visually dissimilar. It was proposed that global and local processing aids the selection of perceptual attributes of objects that are diagnostic for recognition and that selection is based on two mechanisms: spatial extent and grouping/parsing operations. PMID- 17076352 TI - Sequential effects in successive ratio estimation. AB - Sequential effects are examined in four successive ratio estimation (RE) experiments. The procedure in successive RE is identical to that for magnitude estimation (ME), but the task in successive RE is to estimate the ratio of the current to the previous sensation magnitude, and not the separate magnitudes of the sensations. A positive stimulus context effect was found in successive RE for several continua, in agreement with results previously found for ME. The residual autocorrelation for successive RE was zero in many cases, but in some cases negative autocorrelation was found, which is in contrast to the positive autocorrelation that is typically found for ME and other magnitude scaling procedures. It is shown that, when the role of perceptual error is recognized, negative autocorrelation is predicted by a classical model of ratio estimation. Some aspects of response bias are also discussed. PMID- 17076353 TI - Perception of heading without retinal optic flow. AB - How do we determine where we are heading during visually controlled locomotion? Psychophysical research has shown that humans are quite good at judging their travel direction, or heading, from retinal optic flow. Here we show that retinal optic flow is sufficient, but not necessary, for determining heading. By using a purely cyclopean stimulus (random dot cinematogram), we demonstrate heading perception without retinal optic flow. We also show that heading judgments are equally accurate for the cyclopean stimulus and a conventional optic flow stimulus, when the two are matched for motion visibility. The human visual system thus demonstrates flexible, robust use of available visual cues for perceiving heading direction. PMID- 17076354 TI - [Central congenital hypothyroidism due to Graves' disease in the mother]. AB - Two male twins were born at a gestational age of 30 weeks. Five days after delivery, the mother was diagnosed with Graves' disease. The thyroid function in the neonates was therefore evaluated, which led to the detection of central congenital hypothyroidism (central CHT), even though the neonatal CHT-screening had been reported to be normal. Both boys were treated with thyroxine up to the age of nine months. It was then established that their development had been uneventful. Maternal Graves' disease can, due to the presence of anti-thyroid stimulating hormone (TSH) receptor antibodies and the maternal use of anti thyroid drugs, result in thyroid dysfunction in the neonate. Neonates born to mothers with Graves' disease are at risk of developing central CHT. This occurs especially in children of mothers who are not treated or are inadequately treated during pregnancy. In view of the importance of thyroid hormone for brain development, children with central CHT are at risk for neurodevelopmental problems if thyroid dysfunction is not detected and treated early. The Dutch screening for congenital hypothyroidism is based on thyroxine (T4), TSH and thyroid-binding globulin. This makes it possible to detect central CHT. However, in prematurely born infants this disease may be missed because in this subgroup, referral is only based on increased TSH levels, which may not be present. PMID- 17076355 TI - [The practice guideline 'Diabetes mellitus type 2' (second revision) from the Dutch College of General Practitioners; a response from the perspective of general practice]. AB - The publication of the practice guideline 'Diabetes mellitus' by the Dutch College of General Practitioners in 1989 marked the start of an era of publication of several guidelines that helped general practitioners using evidence-based medicine in clinical practice; the guidelines also helped to teach students. The second revision of this guideline presents many improvements, especially simplifications in the medication-schedules. However, the new recommendation to use thiazolidines is based on only one large study and has some unpractical aspects. The new guidelines do not mention preventive action, nor advice regarding early detection. Clinical practice has changed in recent years with the introduction of nurses specialising in diabetes and, despite published research on this subject, the guidelines do not give any recommendations for this. What is also missing are national clinical guidelines for doctors specialising in internal medicine. When patients do not respond to treatment according to the general practitioners' guidelines and are referred to a specialist in internal medicine, the treatment is diverse and seems to be doctor dependent. It is important that treatment there be standardised as well so that general practitioners can refer more effectively. PMID- 17076356 TI - [The practice guideline 'Diabetes mellitus type 2' (second revision) from the Dutch College of General Practitioners; a response from the perspective of internal medicine]. AB - The revised guideline 'Diabetes mellitus type 2' contains several improvements. The HbA1C target level has been lowered to 7% or less. The universal first step in oral therapy has become metformin. The target level for the treatment of hypertension is now a systolic pressure below 140 mmHg. Statins should be prescribed to almost every patient. Finally, ACE-inhibitors are now suggested for all patients with microalbuminuria and hypertension. Some choices made in the present guideline are not evidence-based, e.g. the advice to prescribe pioglitazone to patients with both a body mass index above 27 kg/m2 and cardiovascular disease, but without heart failure. Still, in general, the updated guideline is an important document which has been greatly improved in comparison to the former one. PMID- 17076357 TI - [Non-pharmacological treatment of heart failure: implantable cardioverter defibrillator (ICD) and cardiac resynchronisation therapy]. AB - The non-pharmacological therapy of heart failure, in particular an implantable cardioverter defibrillator (ICD) and cardiac resynchronisation therapy or biventricular stimulation, improves symptoms and survival in patients with heart failure. An ICD is indicated in many patients with heart failure following cardiac arrest unless reversible causes are demonstrable. Selected patients with a left ventricular ejection fraction < or = 35% due to either ischaemic (>40 days after a myocardial infarction) or nonischaemic cardiomyopathy are candidates for ICD implantation as the primary prevention of sudden cardiac death. Patients who continue to have severe symptoms despite maximal pharmacotherapy, with a left ventricular ejection fraction < or = 35% and a wide QRS complex, are candidates for cardiac resynchronisation therapy to improve both symptoms and survival. PMID- 17076358 TI - [The role of the high-density lipoprotein (HDL) particle in atherogenesis and potential methods to increase HDL concentrations]. AB - The concentration of high-density lipoprotein (HDL) cholesterol has a strong inverse relationship to the incidence of cardiovascular disease. The protective effect of HDL cholesterol is due not only to its promotion of reverse cholesterol transport from the vascular wall to the liver, but also to its anti-inflammatory, antioxidative and antithrombotic effects. Patients with low HDL cholesterol concentrations are at increased risk for cardiovascular disease and may be considered for treatment with lipid-lowering drugs, such as statins, niacin and fibrates. Currently, only a limited number of HDL-increasing agents have demonstrated beneficial effects of increasing HDL in studies with intermediary endpoints. Among these drugs are apolipoprotein A-1 variants, inhibitors of the cholesterol ester transfer protein JTT-705 and torcetrapib. PMID- 17076359 TI - [Summary of the practice guideline 'Diabetes mellitus type 2' (second revision) from the Dutch College of General Practitioners]. AB - The practice guideline 'Diabetes mellitus type 2' (second revision) addresses the diagnosis, treatment and management of adults with diabetes mellitus type 2 in general practice. The aim of management is the prevention and treatment of diabetes-related symptoms and complications such as cardiovascular disease, nephro-, retino- and neuropathy. The general practitioner gives the patient education and lifestyle advice and repeats this regularly. In addition, the general practitioner and the patient strive to achieve good glycaemic control. The agent of first choice in the medicinal treatment of all type 2 diabetic patients is metformin. This is continued even after the addition of a sulphonylurea derivative or insulin. This represents a change compared to the previous version of the practice guideline. The indications for thiazolidinediones are limited. To reduce the cardiovascular risk, it is advised to aim at a systolic blood pressure below 140 mmHg. It is also recommended that each patient be prescribed a statin, unless the patient belongs to a subgroup in which the indication for cholesterol lowering therapy is weak or the patient refuses it. Compared to the former guideline, more emphasis is placed on the prevention of nephropathy. The general practitioner is advised to calculate the creatinine clearance yearly and to test for relevant albuminuria in each patient with a life expectancy of 10 years or more. If microalbuminuria is present, the patient is prescribed an angiotensin converting enzyme (ACE) inhibitor, even if the blood pressure is not elevated. The detection of patients with a high risk of diabetic ulcer is also given more emphasis. PMID- 17076360 TI - [Diagnostic image (295). A man with unilateral recurring sinusitis]. AB - A 45-year-old man was seen with recurring sinusitis on the left side due to corpora aliena of glass based on a trauma 25 years before. PMID- 17076361 TI - [Endarterectomy is a good treatment option in patients with asymptomatic carotid stenosis]. AB - Among patients with substantial carotid artery stenosis without neurological symptoms, the benefit of carotid endarterectomy had been unclear until this issue was addressed in two randomised trials. The incidence of stroke during follow-up was reduced by approximately one-half after endarterectomy. In appropriate patients, endarterectomy ofa significant asymptomatic carotid stenosis is certainly a treatment option. PMID- 17076362 TI - [Endarterectomy should not be offered routinely to patients with asymptomatic carotid stenosis]. AB - A recent trial revealed a reduction in ischaemic infarcts after carotid endarterectomy in patients with asymptomatic carotid stenosis. However, the number needed to treat (NNT) was 19 to prevent one stroke in 5 years, a modest effect in comparison to the trials with symptomatic patients (a NNT of only 6 in patients with a symptomatic stenosis of 70-99%). Furthermore, the number of surgical complications outweighs the benefits of surgery during the first two years after treatment. Finally, the benefits also included the prevention of contralateral strokes, whereas it is unlikely that these are prevented by endarterectomy. Therefore, endarterectomy should not be performed routinely in asymptomatic persons. PMID- 17076363 TI - [Combined liver and kidney transplantation: indications and results at the University Medical Centre Groningen, 1994-2005]. AB - OBJECTIVE. To describe the experience with combined liver and kidney transplantation at the University Medical Centre Groningen, The Netherlands. DESIGN. Retrospective. METHOD: Data were analysed from all patients who underwent combined liver and kidney transplantation in the University Medical Centre Groningen, in the period November 1994-December 2005. RESULTS: During the study period 582 orthotopic liver transplantations and 1026 isolated kidney transplantations were performed. 16 patients underwent combined liver and kidney transplantation: 4 were children (aged 17 months-16 years) and 12 were adults (aged 19-59 years). For all patients, both organs were obtained from the same post-mortem donor. Indications for combined liver and kidney transplantation were primary hyperoxaluria type I (n=6), polycystic liver and kidney disease (n=3) and unrelated liver and kidney failure (n=7). The 1- and 5-year survival rate was 88% (14/16), which was not significantly different from the results after isolated liver transplantation. Two patients died 11 days and 74 months after combined transplantation, due to complications from unsuccessful retransplantation of the liver for hepatic artery thrombosis and secondary biliary cirrhosis, respectively. A third patient died 51 days after combined transplantation due to sepsis. CONCLUSION: Combined liver and kidney transplantation was a life-saving intervention in this selected group of patients with combined liver and kidney failure. Patient survival was comparable to that of patients undergoing isolated liver transplantation. PMID- 17076364 TI - [Gastroenteritis caused by Salmonella from pet snakes]. AB - A Salmonella subspecies associated with reptiles (Salmonella enterica subspecies diarizonae) was isolated from the stool of a 19-year-old man with gastroenteritis. The same species was isolated from stool and urine samples taken from terraria found in the home of the patient's parents where snakes were kept. A high percentage of reptiles in the wild and in captivity are asymptomatic carriers of Salmonella species that can be transmitted to humans who come in contact with these animals. Unlike in the United States of America, for example, cases of reptile-associated infections have scarcely been published in the Netherlands and targeted information on the risk of infection is lacking. Because the popularity of exotic pets--and thereby the risk of infection--is increasing in The Netherlands, targeted information for veterinarians, traders and owners of exotic pets is warranted to prevent reptile-associated salmonellosis. PMID- 17076365 TI - [Gastrojejunostomy versus endoscopic stent placement as palliative treatment for a malignant constriction of the duodenum: the SUSTENT-study]. AB - Gastric-outlet obstruction often results from inoperable distal stomach, periampullary (pancreatic or cholangio-), or duodenal carcinoma. Gastrojejunostomy and stent placement are standard palliative treatments. An advantage of gastrojejunostomy is the long-term efficacy; a disadvantage is the prolonged postoperative recovery time. The advantage of stent placement is the rapid ability to consume a soft diet; a disadvantage is that around 20% of the patients require re-intervention because of recurrent symptoms. A randomised multicentre study was started in January 2006 in the Netherlands in which gastrojejunostomy is compared with stent placement in the palliative treatment of malignant gastroduodenal obstruction: 'Surgery versus stent for malignant gastroduodenal obstruction', the SUSTENT-study. The primary-outcome measurement is survival adjusted for the time patients are not able to consume (soft) food. Other outcome measurements are medical effects (complications, re-interventions), quality of life, cost and cost-effectiveness. This study aims to provide individualised recommendations for effective palliative treatment of patients with malignant gastroduodenal obstruction. PMID- 17076366 TI - [Advantages of working in a developing country during medical specialty training]. AB - Training in a developing country provides additional value for Dutch physicians who are training for a medical specialty. Knowledge of and experience with tropical diseases is also important in the Netherlands. The limited access to diagnostic tests and treatments in developing countries forces physicians to perform physical examinations meticulously and prescribe treatments conscientiously. Limited abilities to communicate can lead to an appreciation of the importance of communication and insights regarding optimal methods for communication. By working with staff in developing countries, physicians learn to be flexible in regard to work attitudes and cooperation. Physicians also learn to develop a personal academic programme independently. In addition, physicians become more readily involved in improving standards of living and healthcare in the community as a whole, and in the coordination and organization of work. Lastly, the ability to work professionally with limitations in healthcare is also valuable in medical practice in the Netherlands. Therefore, the decision to do part of the training as a medical specialist in a developing country should be supported. This experience should also count as part of the training. PMID- 17076367 TI - [Sponsoring of research by the industry is not good for science]. PMID- 17076368 TI - [Double-blind, placebo-controlled cow's milk challenge in children with alleged cow's milk allergies, performed in a general hospital: diagnosis rejected in two thirds of the children]. PMID- 17076369 TI - [Hypoglycemic coma due to falsely elevated glucose values in a patient with diabetes mellitus and peritoneal dialysis]. PMID- 17076370 TI - Catching your breath: research efforts to analyze the negative effects of air pollution on human health. PMID- 17076371 TI - Performance reporting and public health: bridging the gap. PMID- 17076372 TI - Variation in heart attack mortality in Canada. PMID- 17076373 TI - In conversation with George Smitherman. Interview by Ken Tremblay. PMID- 17076374 TI - Timely and appropriate access to healthcare. The way forward--the roles of the players. AB - Through the leadership of a number of Canadian healthcare organizations and the Canadian Policy Research Networks, invitational conferences focused on the "wait time problem" have been held in Ottawa over the past three years. This paper, utilizing both the views and information presented at the most recent conference and the author's own experience and views, seeks to outline directions that could help us achieve timely and appropriate access to care in Canada. PMID- 17076375 TI - The pivotal role of critical care and surgical efficiencies in supporting Ontario's Wait Time Strategy: Part 3. AB - As part of its Wait Time Strategy, the Ontario Ministry of Health and Long-Term Care provided significant amounts of money to perform more cases with the understanding that improving access by reducing wait times is not just a matter of increasing funding. Rather, fundamental system and practice change is required to sustain improvements in the long term. PMID- 17076376 TI - Paying for institutional quality: reaching the limits of funding policy? AB - As a recent issue of the Longwoods journal, HealthcarePapers (Vol. 6, No. 4), indicates, interest in pay-for-performance incentives is on the increase in both the US and Canada. Weighing in on the topic, the authors suggest that the heightened emphasis on quality and patient safety are two primary drivers. PMID- 17076377 TI - Has Ontario's Stroke System really made a difference? AB - The progress that has been made in preventing and treating stroke since 2000, when the Joint Stroke Strategy Working Group tabled its blueprint report, Towards an Integrated Stroke Strategy, is very encouraging. The evaluation results demonstrate that the Ontario Stroke System has had positive measurable impacts on access to stroke-related services, the integration and coordination of stroke care, treatment for stroke, and client and provider satisfaction. PMID- 17076378 TI - Achieving team consensus on best practice rehabilitation guidelines following primary total hip replacement (THR) surgery. AB - The demand for hip replacements is on the rise; however, despite commonly employed post-operative positional and activity restrictions, limited evidence exists to substantiate such practices. Guidelines provide a means of assisting the healthcare team in decision-making; however, when a clinical decision has to be made in the absence of scientific evidence, guideline developers rely on consensus processes. A formal consensus process based on a nominal group technique was used to incorporate the scientific evidence and practitioner experience for the development of primary total hip replacement guidelines at a specialized orthopaedic tertiary care facility. Agreement was reached regarding a number of practice issues, which historically had varied among the surgeons. Given the lack of available guidance on how to synthesize research findings in conjunction with expert opinion, the process we employed has been described so that others may use, modify and refine it. Further scientific inquiry into safe activity resumption post-total hip replacement is warranted. PMID- 17076379 TI - Evaluating organizational readiness for change: a preliminary mixed-model assessment of an interprofessional rehabilitation hospital. AB - We conducted a Functional Organizational Readiness for Change Evaluation (FORCE) to assess the characteristics of readiness for change across two programs (N=216 employees) in an interprofessional rehabilitation hospital that was about to undergo strategic changes as part of a planned physical merger within the next two years. The study used a mixed-method approach: a quantitative survey, previously validated in a drug rehabilitation setting, followed by key informant interviews to further enlighten survey findings. Statistical analyses identified correlations between demographic variables (age, education and experience) and readiness for change, as well as the prevalence of specific organizational characteristics (motivation for change, access to resources, staff attributes, organizational climate, and exposure/ use of training opportunities) that facilitate or impede change. Findings were intended to better inform the tactics for successful implementation of upcoming initiatives. Much like assessing a patient prior to initiating a treatment, FORCE can serve as a management tool to direct the planning and implementation of changes intended to improve hospital performance. PMID- 17076380 TI - Opening the doors: building brand awareness. AB - In search of a credible and cost-effective way to promote Mount Sinai Hospital (Mount Sinai) and educate a broad public, Mount Sinai opened its doors wide to The Globe and Mail (The Globe). The result was a three-part national feature series that told Mount Sinai's compelling story, provided third-party credibility and confirmed the value of proactive media relations. The series engaged our staff, energized our volunteers and reached both stakeholders and the general public on an emotional level. It also generated the more than dollars 6 million worth of equivalent advertising space it would have required for Mount Sinai to reach this national audience. PMID- 17076381 TI - Cross border shopping: how much for your heart? AB - No one has described the fiscal disparity that arises as a result of Canada/United States (US) cross border organ transactions. Canadian healthcare cannot continue to subsidize US transplant programs while paying their inflated organ acquisition fees. Canadian hospitals should bill US transplant programs for all costs associated with procuring organs. PMID- 17076382 TI - Clinical electronic communications: a new paradigm that is here to stay? AB - The authors discuss the current status of clinical electronic communication, including e-mail and secure Web messaging, among physicians and between physicians and their patients. The paper provides a concise summary of what is known about clinical electronic communications in healthcare: what is working, what the challenges are and what is promising. PMID- 17076383 TI - Low-cost rapid usability engineering: designing and customizing usable healthcare information systems. AB - It is essential that the healthcare systems we develop are usable, meet user information needs and are safe. To ensure system usability, a variety of methods have emerged from the area of usability engineering and have been adapted to healthcare. The authors have been applying methods of usability engineering, working with hospitals and companies to develop more usable healthcare information systems for over 15 years. Based on our current work at the University of Victoria, we describe how to set up a low-cost portable laboratory that can rapidly evaluate the usability and safety of healthcare information systems both in artificial mocked-up settings and in real clinical contexts (e.g., in hospital wards). PMID- 17076384 TI - Medical transcription outsourcing. PMID- 17076385 TI - Communication and contraception in rural Bangladesh. AB - This paper examines the association of communication in explaining the decision of women in rural Bangladesh to use or not to use contraception. Using survey data from villages in Bangladesh, we found that communication is an important influence on the ideational change for a smaller family norm and the practice of contraception. This is evident even when socioeconomic and cultural variables are controlled for. We recommend that to foster the use of contraception, communication factors should be given greater emphasis, especially to target husbands in the family planning program and to improve the image of female children. PMID- 17076386 TI - New computing model helps Hamilton Health Sciences address changing business requirements. AB - This case study presents the impetus, business case, chronology and benefits of implementing a new server-based computing model at Hamilton Health Sciences that solved a critical desktop management problem while reducing IT costs. The new approach also provided a robust, flexible and scalable technology platform that is helping the hospital address business requirements driven by the emerging virtual healthcare community. PMID- 17076387 TI - Error-resilient video communications over CDMA networks with a bandwidth constraint. AB - We present an adaptive video transmission scheme for use in a code-division multiple-access network, which incorporates efficient bandwidth allocation among source coding, channel coding, and spreading under a fixed total bandwidth constraint. We derive the statistics of the received signal, as well as a theoretical bound on the packet drop rate at the receiver. Based on these results, a bandwidth allocation algorithm is proposed at the packet level, which incorporates the effects of both the changing channel conditions and the dynamics of the source content. Detailed simulations are done to evaluate the performance of the system, and the sensitivity of the system to estimation error is presented. PMID- 17076388 TI - On the use of context-weighting in lossless bilevel image compression. AB - We present a context-weighting algorithm that adaptively weights in real-time three-context models based on their relative accuracy. It can automatically select the better model over different regions of an image, producing better probability estimates than using either one of these models exclusively. Combined with the previously proposed block arithmetic coder for image compression (BACIC), the overall performance is slightly better than JBIG for the eight CCITT business-type test images, outperforms JBIG by 13.8% on halftone images, and by 17.5% for compounded images containing both text and halftones. Furthermore, users no longer need to select models as in JBIG and BACIC to get the better performance. PMID- 17076389 TI - Reversing demosaicking and compression in color filter array image processing: performance analysis and modeling. AB - In the conventional processing chain of single-sensor digital still cameras (DSCs), the images are captured with color filter arrays (CFAs) and the CFA samples are demosaicked into a full color image before compression. To avoid additional data redundancy created by the demosaicking process, an alternative processing chain has been proposed to move the compression process before the demosaicking. Recent empirical studies have shown that the alternative chain can outperform the conventional one in terms of image quality at low compression ratios. To provide a theoretically sound basis for such conclusion, we propose analytical models for the reconstruction errors of the two processing chains. The models developed confirm the results of existing empirical studies and provide better understanding of DSC processing chains. The modeling also allows performance predictions for more advanced compression and demosaicking methods, thus providing important cues for future development in this area. PMID- 17076390 TI - Precompression quality-control algorithm for JPEG 2000. AB - In this paper, a precompression quality-control algorithm is proposed. It can greatly reduce computational power of the embedded block coding (EBC) and memory requirement to buffer bit streams. By using the propagation property and the randomness property of the EBC algorithm, rate and distortion of coding passes is approximately predicted. Thus, the truncation points are chosen before actual coding by the entropy coder. Therefore, the computational power, which is measured with the number of contexts to be processed, is greatly reduced since most of the computations are skipped. The memory requirement, which is measured with the amount required to buffer bit streams, is also reduced since the skipped contexts do not generate bit streams. Experimental results show that the proposed algorithm reduces the computational power of the EBC by 80% on average at 0.8 bpp compared with the conventional postcompression rate-distortion optimization algorithm. Moreover, the memory requirement is also reduced by 90%. The average PSNR degrades only about 0.1-0.3 dB, on average. PMID- 17076391 TI - Quadratic weighted median filters for edge enhancement of noisy images. AB - Quadratic Volterra filters are effective in image sharpening applications. The linear combination of polynomial terms, however, yields poor performance in noisy environments. Weighted median (WM) filters, in contrast, are well known for their outlier suppression and detail preservation properties. The WM sample selection methodology is naturally extended to the quadratic sample case, yielding a filter structure referred to as quadratic weighted median (QWM) that exploits the higher order statistics of the observed samples while simultaneously being robust to outliers arising in the higher order statistics of environment noise. Through statistical analysis of higher order samples, it is shown that, although the parent Gaussian distribution is light tailed, the higher order terms exhibit heavy-tailed distributions. The optimal combination of terms contributing to a quadratic system, i.e., cross and square, is approached from a maximum likelihood perspective which yields the WM processing of these terms. The proposed QWM filter structure is analyzed through determination of the output variance and breakdown probability. The studies show that the QWM exhibits lower variance and breakdown probability indicating the robustness of the proposed structure. The performance of the QWM filter is tested on constant regions, edges and real images, and compared to its weighted-sum dual, the quadratic Volterra filter. The simulation results show that the proposed method simultaneously suppresses the noise and enhances image details. Compared with the quadratic Volterra sharpener, the QWM filter exhibits superior qualitative and quantitative performance in noisy image sharpening. PMID- 17076392 TI - The multidimensional phase unwrapping integral and applications to microwave tomographical image reconstruction. AB - Spatial unwrapping of the phase component of time varying electromagnetic fields has important implications in a range of disciplines including synthetic aperture radar (SAR) interferometry, MRI, optical confocal microscopy, and microwave tomography. This paper presents a fundamental framework based on the phase unwrapping integral, especially in the complex case where phase singularities are enclosed within the closed path integral. With respect to the phase unwrapping required when utilized in Gauss-Newton iterative microwave image reconstruction, the concept of dynamic phase unwrapping is introduced where the singularity location varies as a function of the iteratively modified property distributions. Strategies for dynamic phase unwrapping in the microwave problem were developed and successfully tested in simulations and clinical experiments utilizing large, high contrast targets to validate the approach. PMID- 17076393 TI - An improved observation model for super-resolution under affine motion. AB - Super-resolution (SR) techniques make use of subpixel shifts between frames in an image sequence to yield higher resolution images. We propose an original observation model devoted to the case of nonisometric inter-frame motion as required, for instance, in the context of airborne imaging sensors. First, we describe how the main observation models used in the SR literature deal with motion, and we explain why they are not suited for nonisometric motion. Then, we propose an extension of the observation model by Elad and Feuer adapted to affine motion. This model is based on a decomposition of affine transforms into successive shear transforms, each one efficiently implemented by row-by-row or column-by-column one-dimensional affine transforms. We demonstrate on synthetic and real sequences that our observation model incorporated in a SR reconstruction technique leads to better results in the case of variable scale motions and it provides equivalent results in the case of isometric motions. PMID- 17076394 TI - Fast IIR isotropic 2-D complex Gabor filters with boundary initialization. AB - Gabor filters are widely applied in image analysis and computer vision applications. This paper describes a fast algorithm for isotropic complex Gabor filtering that outperforms existing implementations. The main computational improvement arises from the decomposition of Gabor filtering into more efficient Gaussian filtering and sinusoidal modulations. Appropriate filter initial conditions are derived to avoid boundary transients, without requiring explicit image border extension. Our proposal reduces up to 39% the number of required operations with respect to state-of-the-art approaches. A full C++ implementation of the method is publicly available. PMID- 17076395 TI - Face verification across age progression. AB - Human faces undergo considerable amounts of varialions with aging. While face recognition systems have been proven to be sensitive to factors such as illumination and pose, their sensitivity to facial aging effects is yet to be studied. How does age progression affect the similarity between a pair of face images of an individual? What is the confidence associated with establishing the identity between a pair of age separated face images? In this paper, we develop a Bayesian age difference classifier that classifies face images of individuals based on age differences and performs face verification across age progression. Further, we study the similarity of faces across age progression. Since age separated face images invariably differ in illumination and pose, we propose preprocessing methods for minimizing such variations. Experimental results using a database comprising of pairs of face images that were retrieved from the passports of 465 individuals are presented. The verification system for faces separated by as many as nine years, attains an equal error rate of 8.5%. PMID- 17076396 TI - Translation-invariant contourlet transform and its application to image denoising. AB - Most subsampled filter banks lack the feature of translation invariance, which is an important characteristic in denoising applications. In this paper, we study and develop new methods to convert a general multichannel, multidimensional filter bank to a corresponding translation-invariant (TI) framework. In particular, we propose a generalized algorithme a trous, which is an extension of the algorithme a trous introduced for 1-D wavelet transforms. Using the proposed algorithm, as well as incorporating modified versions of directional filter banks, we construct the TI contourlet transform (TICT). To reduce the high redundancy and complexity of the TICT, we also introduce semi-translation invariant contourlet transform (STICT). Then, we employ an adapted bivariate shrinkage scheme to the STICT to achieve an efficient image denoising approach. Our experimental results demonstrate the benefits and potential of the proposed denoising approach. Complexity analysis and efficient realization of the proposed TI schemes are also presented. PMID- 17076397 TI - Fast splitting alpha-rooting method of image enhancement: tensor representation. AB - In the tensor representation, a two-dimensional (2-D) image is represented uniquely by a set of one-dimensional (1-D) signals, so-called splitting-signals, that carry the spectral information of the image at frequency-points of specific sets that cover the whole domain of frequencies. The image enhancement is thus reduced to processing splitting-signals and such process requires a modification of only a few spectral components of the image, for each signal. For instance, the alpha-rooting method of image enhancement can be fulfilled through processing separately a maximum of 3N/2 splitting-signals of an image (N x N), where N is a power of two. In this paper, we propose a fast implementation of the a-rooting method by using one splitting-signal of the tensor representation with respect to the discrete Fourier transform (DFT). The implementation is described in the frequency and spatial domains. As a result, the proposed algorithms for image enhancement use two 1-D N-point DFTs instead of two 2-D N x N-point DFTs in the traditional method of alpha-rooting. PMID- 17076398 TI - Multiresolution MAP despeckling of SAR images based on locally adaptive generalized Gaussian pdf modeling. AB - In this paper, a new despeckling method based on undecimated wavelet decomposition and maximum a posteriori MIAP) estimation is proposed. Such a method relies on the assumption that the probability density function (pdf) of each wavelet coefficient is generalized Gaussian (GG). The major novelty of the proposed approach is that the parameters of the GG pdf are taken to be space varying within each wavelet frame. Thus, they may be adjusted to spatial image context, not only to scale and orientation. Since the MAP equation to be solved is a function of the parameters of the assumed pdf model, the variance and shape factor of the GG function are derived from the theoretical moments, which depend on the moments and joint moments of the observed noisy signal and on the statistics of speckle. The solution of the MAP equation yields the MAP estimate of the wavelet coefficients of the noise-free image. The restored SAR image is synthesized from such coefficients. Experimental results, carried out on both synthetic speckled images and true SAR images, demonstrate that MAP filtering can be successfully applied to SAR images represented in the shift-invariant wavelet domain, without resorting to a logarithmic transformation. PMID- 17076399 TI - Ground target recognition using rectangle estimation. AB - We propose a ground target recognition method based on 3-D laser radar data. The method handles general 3-D scattered data. It is based on the fact that man-made objects of complex shape can be decomposed to a set of rectangles. The ground target recognition method consists of four steps; 3-D size and orientation estimation, target segmentation into parts of approximately rectangular shape, identification of segments that represent the target's functional/main parts, and target matching with CAD models. The core in this approach is rectangle estimation. The performance of the rectangle estimation method is evaluated statistically using Monte Carlo simulations. A case study on tank recognition is shown, where 3-D data from four fundamentally different types of laser radar systems are used. Although the approach is tested on rather few examples, we believe that the approach is promising. PMID- 17076400 TI - The Bayesian operating point of the Canny edge detector. AB - We have investigated the operating point of the Canny edge detector which minimizes the Bayes risk of misclassification. By considering each of the sequential stages which constitute the Canny algorithm, we conclude that the linear filtering stage of Canny, without postprocessing, performs very poorly by any standard in pattern recognition and achieves error rates which are almost indistinguishable from a priori classification. We demonstrate that the edge detection performance of the Canny detector is due almost entirely to the postprocessing stages of nonmaximal suppression and hysteresis thresholding. PMID- 17076401 TI - Recognition of dynamic video contents with global probabilistic models of visual motion. AB - The exploitation of video data requires methods able to extract high-level information from the images. Video summarization, video retrieval, or video surveillance are examples of applications. In this paper, we tackle the challenging problem of recognizing dynamic video contents from low-level motion features. We adopt a statistical approach involving modeling, (supervised) learning, and classification issues. Because of the diversity of video content (even for a given class of events), we have to design appropriate models of visual motion and learn them from videos. We have defined original parsimonious global probabilistic motion models, both for the dominant image motion (assumed to be due to the camera motion) and the residual image motion (related to scene motion). Motion measurements include affine motion models to capture the camera motion and low-level local motion features to account for scene motion. Motion learning and recognition are solved using maximum likelihood criteria. To validate the interest of the proposed motion modeling and recognition framework, we report dynamic content recognition results on sports videos. PMID- 17076402 TI - Unsupervised variational image segmentation/classification using a Weibull observation model. AB - Studies have shown that the Weibull distribution can model accurately a wide variety of images. Its parameters index a family of distributions which includes the exponential and approximations of the Gaussian and the Raleigh models widely used in image segmentation. This study investigates the Weibull distribution in unsupervised image segmentation and classification by a variational method. The data term of the segmentation functional measures the conformity of the image intensity in each region to a Weibull distribution whose parameters are determined jointly with the segmentation. Minimization of the functional is implemented by active curves via level sets and consists of iterations of two consecutive steps: curve evolution via Euler-Lagrange descent equations and evaluation of the Weibull distribution parameters. Experiments with synthetic and real images are described which verify the validity of method and its implementation. PMID- 17076403 TI - A statistical evaluation of recent full reference image quality assessment algorithms. AB - Measurement of visual quality is of fundamental importance for numerous image and video processing applications, where the goal of quality assessment (QA) algorithms is to automatically assess the quality of images or videos in agreement with human quality judgments. Over the years, many researchers have taken different approaches to the problem and have contributed significant research in this area and claim to have made progress in their respective domains. It is important to evaluate the performance of these algorithms in a comparative setting and analyze the strengths and weaknesses of these methods. In this paper, we present results of an extensive subjective quality assessment study in which a total of 779 distorted images were evaluated by about two dozen human subjects. The "ground truth" image quality data obtained from about 25,000 individual human quality judgments is used to evaluate the performance of several prominent full-reference image quality assessment algorithms. To the best of our knowledge, apart from video quality studies conducted by the Video Quality Experts Group, the study presented in this paper is the largest subjective image quality study in the literature in terms of number of images, distortion types, and number of human judgments per image. Moreover, we have made the data from the study freely available to the research community. This would allow other researchers to easily report comparative results in the future. PMID- 17076404 TI - Perceptual image hashing via feature points: performance evaluation and tradeoffs. AB - We propose an image hashing paradigm using visually significant feature points. The feature points should be largely invariant under perceptually insignificant distortions. To satisfy this, we propose an iterative feature detector to extract significant geometry preserving feature points. We apply probabilistic quantization on the derived features to introduce randomness, which, in turn, reduces vulnerability to adversarial attacks. The proposed hash algorithm withstands standard benchmark (e.g., Stirmark) attacks, including compression, geometric distortions of scaling and small-angle rotation, and common signal processing operations. Content changing (malicious) manipulations of image data are also accurately detected. Detailed statistical analysis in the form of receiver operating characteristic (ROC) curves is presented and reveals the success of the proposed scheme in achieving perceptual robustness while avoiding misclassification. PMID- 17076405 TI - Structure from planar motion. AB - Planar motion is arguably the most dominant type of motion in surveillance videos. The constraints on motion lead to a simplified factorization method for structure from planar motion when using a stationary perspective camera. Compared with methods for general motion, our approach has two major advantages: a measurement matrix that fully exploits the motion constraints is formed such that the new measurement matrix has a rank of at most 3, instead of 4; the measurement matrix needs similar scalings, but the estimation of fundamental matrices or epipoles is not needed. Experimental results show that the algorithm is accurate and fairly robust to noise and inaccurate calibration. As the new measurement matrix is a nonlinear function of the observed variables, a different method is introduced to deal with the directional uncertainty in the observed variables. Differences and the dual relationship between planar motion and planar object are also clarified. Based on our method, a fully automated vehicle reconstruction system has been designed. PMID- 17076406 TI - On the orientability of shapes. AB - The orientation of a shape is a useful quantity, and has been shown to affect performance of object recognition in the human visual system. Shape orientation has also been used in computer vision to provide a properly oriented frame of reference, which can aid recognition. However, for certain shapes, the standard moment-based method of orientation estimation fails. We introduce as a new shape feature shape orientability, which defines the degree to which a shape has distinct (but not necessarily unique) orientation. A new method is described for measuring shape orientability, and has several desirable properties. In particular, unlike the standard moment-based measure of elongation, it is able to differentiate between the varying levels of orientability of n-fold rotationally symmetric shapes. Moreover, the new orientability measure is simple and efficient to compute (for an n-gon we describe an O(n) algorithm). PMID- 17076407 TI - A wavelet-based two-stage near-lossless coder. AB - In this paper, we present a two-stage near-lossless compression scheme. It belongs to the class of "lossy plus residual coding" and consists of a wavelet based lossy layer followed by arithmetic coding of the quantized residual to guarantee a given L(infinity) error bound in the pixel domain. We focus on the selection of the optimum bit rate for the lossy layer to achieve the minimum total bit rate. Unlike other similar lossy plus lossless approaches using a wavelet-based lossy layer, the proposed method does not require iteration of decoding and inverse discrete wavelet transform in succession to locate the optimum bit rate. We propose a simple method to estimate the optimal bit rate, with a theoretical justification based on the critical rate argument from the rate-distortion theory and the independence of the residual error. PMID- 17076408 TI - An optimal nonorthogonal separation of the anisotropic Gaussian convolution filter. AB - We give an analytical and geometrical treatment of what it means to separate a Gaussian kernel along arbitrary axes in R(n), and we present a separation scheme that allows us to efficiently implement anisotropic Gaussian convolution filters for data of arbitrary dimensionality. Based on our previous analysis we show that this scheme is optimal with regard to the number of memory accesses and interpolation operations needed. The proposed method relies on nonorthogonal convolution axes and works completely in image space. Thus, it avoids the need for a fast Fourier transform (FFT)-subroutine. Depending on the accuracy and speed requirements, different interpolation schemes and methods to implement the one-dimensional Gaussian (finite impulse response and infinite impulse response) can be integrated. Special emphasis is put on analyzing the performance and accuracy of the new method. In particular, we show that without any special optimization of the source code, it can perform anisotropic Gaussian filtering faster than methods relying on the FFT. PMID- 17076409 TI - Embedding motion in model-based stochastic tracking. AB - Particle filtering is now established as one of the most popular methods for visual tracking. Within this framework, there are two important considerations. The first one refers to the generic assumption that the observations are temporally independent given the sequence of object states. The second consideration, often made in the literature, uses the transition prior as the proposal distribution. Thus, the current observations are not taken into account, requiring the noise process of this prior to be large enough to handle abrupt trajectory changes. As a result, many particles are either wasted in low likelihood regions of the state space, resulting in low sampling efficiency, or more importantly, propagated to distractor regions of the image, resulting in tracking failures. In this paper, we propose to handle both considerations using motion. We first argue that, in general, observations are conditionally correlated, and propose a new model to account for this correlation, allowing for the natural introduction of implicit and/or explicit motion measurements in the likelihood term. Second, explicit motion measurements are used to drive the sampling process towards the most likely regions of the state space. Overall, the proposed model handles abrupt motion changes and filters out visual distractors, when tracking objects with generic models based on shape or color distribution. Results were obtained on head tracking experiments using several sequences with moving camera involving large dynamics. When compared against the Condensation Algorithm, they have demonstrated the superior tracking performance of our approach. PMID- 17076410 TI - Building the component tree in quasi-linear time. AB - The level sets of a map are the sets of points with level above a given threshold. The connected components of the level sets, thanks to the inclusion relation, can be organized in a tree structure, that is called the component tree. This tree, under several variations, has been used in numerous applications. Various algorithms have been proposed in the literature for computing the component tree. The fastest ones (considering the worst-case complexity) have been proven to run in O(n ln(n)). In this paper, we propose a simple to implement quasi-linear algorithm for computing the component tree on symmetric graphs, based on Tarjan's union-find procedure. We also propose an algorithm that computes the n most significant lobes of a map. PMID- 17076411 TI - Velocity-image model for online signature verification. AB - In general, online signature capturing devices provide outputs in the form of shape and velocity signals. In the past, strokes have been extracted while tracking velocity signal minimas. However, the resulting strokes are larger and complicated in shape and thus make the subsequent job of generating a discriminative template difficult. We propose a new stroke-based algorithm that splits velocity signal into various bands. Based on these bands, strokes are extracted which are smaller and more simpler in nature. Training of our proposed system revealed that low- and high-velocity bands of the signal are unstable, whereas the medium-velocity band can be used for discrimination purposes. Euclidean distances of strokes extracted on the basis of medium velocity band are used for verification purpose. The experiments conducted show improvement in discriminative capability of the proposed stroke-based system. PMID- 17076412 TI - Binary tree-based generic demosaicking algorithm for multispectral filter arrays. AB - In this paper, we extend the idea of using mosaicked color filter array (CFA) in color imaging, which has been widely adopted in the digital color camera industry, to the use of multispectral filter array (MSFA) in multispectral imaging. The filter array technique can help reduce the cost, achieve exact registration, and improve the robustness of the imaging system. However, the extension from CFA to MSFA is not straightforward. First, most CFAs only deal with a few bands (3 or 4) within the narrow visual spectral region, while the design of MSFA needs to handle the arrangement of multiple bands (more than 3) across a much wider spectral range. Second, most existing CFA demosaicking algorithms assume the fixed Bayer CFA and are confined to properties only existed in the color domain. Therefore, they cannot be directly applied to multispectral demosaicking. The main challenges faced in multispectral demosaicking is how to design a generic algorithm that can handle the more diversified MSFA patterns, and how to improve performance with a coarser spatial resolution and a less degree of spectral correlation. In this paper, we present a binary tree based generic demosaicking method. Two metrics are used to evaluate the generic algorithm, including the root mean-square error (RMSE) for reconstruction performance and the classification accuracy for target discrimination performance. Experimental results show that the demosaicked images present low RMSE (less than 7) and comparable classification performance as original images. These results support that MSFA technique can be applied to multispectral imaging with unique advantages. PMID- 17076413 TI - Adaptive segmentation of textured images by using the coupled Markov random field model. AB - Although simple and efficient, traditional feature-based texture segmentation methods usually suffer from the intrinsical less inaccuracy, which is mainly caused by the oversimplified assumption that each textured subimage used to estimate a feature is homogeneous. To solve this problem, an adaptive segmentation algorithm based on the coupled Markov random field (CMRF) model is proposed in this paper. The CMRF model has two mutually dependent components: one models the observed image to estimate features, and the other models the labeling to achieve segmentation. When calculating the feature of each pixel, the homogeneity of the subimage is ensured by using only the pixels currently labeled as the same pattern. With the acquired features, the labeling is obtained through solving a maximum a posteriori problem. In our adaptive approach, the feature set and the labeling are mutually dependent on each other, and therefore are alternately optimized by using a simulated annealing scheme. With the gradual improvement of features' accuracy, the labeling is able to locate the exact boundary of each texture pattern adaptively. The proposed algorithm is compared with a simple MRF model based method in segmentation of Brodatz texture mosaics and real scene images. The satisfying experimental results demonstrate that the proposed approach can differentiate textured images more accurately. PMID- 17076414 TI - Fuzzy two-step filter for impulse noise reduction from color images. AB - A new framework for reducing impulse noise from digital color images is presented, in which a fuzzy detection phase is followed by an iterative fuzzy filtering technique. We call this filter the fuzzy two-step color filter. The fuzzy detection method is mainly based on the calculation of fuzzy gradient values and on fuzzy reasoning. This phase determines three separate membership functions that are passed to the filtering step. These membership functions will be used as a representation of the fuzzy set impulse noise (one function for each color component). Our proposed new fuzzy method is especially developed for reducing impulse noise from color images while preserving details and texture. Experiments show that the proposed filter can be used for efficient removal of impulse noise from color images without distorting the useful information in the image. PMID- 17076415 TI - Spatially variant morphological restoration and skeleton representation. AB - The theory of spatially variant (SV) mathematical morphology is used to extend and analyze two important image processing applications: morphological image restoration and skeleton representation of binary images. For morphological image restoration, we propose the SV alternating sequential filters and SV median filters. We establish the relation of SV median filters to the basic SV morphological operators (i.e., SV erosions and SV dilations). For skeleton representation, we present a general framework for the SV morphological skeleton representation of binary images. We study the properties of the SV morphological skeleton representation and derive conditions for its invertibility. We also develop an algorithm for the implementation of the SV morphological skeleton representation of binary images. The latter algorithm is based on the optimal construction of the SV structuring element mapping designed to minimize the cardinality of the SV morphological skeleton representation. Experimental results show the dramatic improvement in the performance of the SV morphological restoration and SV morphological skeleton representation algorithms in comparison to their translation-invariant counterparts. PMID- 17076416 TI - Recovering DC coefficients in block-based DCT. AB - It is a common approach for JPEG and MPEG encryption systems to provide higher protection for dc coefficients and less protection for ac coefficients. Some authors have employed a cryptographic encryption algorithm for the dc coefficients and left the ac coefficients to techniques based on random permutation lists which are known to be weak against known-plaintext and chosen ciphertext attacks. In this paper we show that in block-based DCT, it is possible to recover dc coefficients from ac coefficients with reasonable image quality and show the insecurity of image encryption methods which rely on the encryption of dc values using a cryptoalgorithm. The method proposed in this paper combines dc recovery from ac coefficients and the fact that ac coefficients can be recovered using a chosen ciphertext attack. We demonstrate that a method proposed by Tang to encrypt and decrypt MPEG video can be completely broken. PMID- 17076417 TI - Multitraining support vector machine for image retrieval. AB - Relevance feedback (RF) schemes based on support vector machines (SVMs) have been widely used in content-based image retrieval (CBIR). However, the performance of SVM-based RF approaches is often poor when the number of labeled feedback samples is small. This is mainly due to 1) the SVM classifier being unstable for small size training sets because its optimal hyper plane is too sensitive to the training examples; and 2) the kernel method being ineffective because the feature dimension is much greater than the size of the training samples. In this paper, we develop a new machine learning technique, multitraining SVM (MTSVM), which combines the merits of the cotraining technique and a random sampling method in the feature space. Based on the proposed MTSVM algorithm, the above two problems can be mitigated. Experiments are carried out on a large image set of some 20,000 images, and the preliminary results demonstrate that the developed method consistently improves the performance over conventional SVM-based RFs in terms of precision and standard deviation, which are used to evaluate the effectiveness and robustness of a RF algorithm, respectively. PMID- 17076418 TI - General logarithmic image processing convolution. AB - The logarithmic image processing model (LIP) is a robust mathematical framework, which, among other benefits, behaves invariantly to illumination changes. This paper presents, for the first time, two general formulations of the 2-D convolution of separable kernels under the LIP paradigm. Although both formulations are mathematically equivalent, one of them has been designed avoiding the operations which are computationally expensive in current computers. Therefore, this fast LIP convolution method allows to obtain significant speedups and is more adequate for real-time processing. In order to support these statements, some experimental results are shown in Section V. PMID- 17076419 TI - Orthogonal laplacianfaces for face recognition. AB - Following the intuition that the naturally occurring face data may be generated by sampling a probability distribution that has support on or near a submanifold of ambient space, we propose an appearance-based face recognition method, called orthogonal Laplacianface. Our algorithm is based on the locality preserving projection (LPP) algorithm, which aims at finding a linear approximation to the eigenfunctions of the Laplace Beltrami operator on the face manifold. However, LPP is nonorthogonal, and this makes it difficult to reconstruct the data. The orthogonal locality preserving projection (OLPP) method produces orthogonal basis functions and can have more locality preserving power than LPP. Since the locality preserving power is potentially related to the discriminating power, the OLPP is expected to have more discriminating power than LPP. Experimental results on three face databases demonstrate the effectiveness of our proposed algorithm. PMID- 17076420 TI - Camera calibration using symmetric objects. AB - This paper proposes a novel method for camera calibration using images of a mirror symmetric object. Assuming unit aspect ratio and zero skew, we show that interimage homographies can be expressed as a function of only the principal point. By minimizing symmetric transfer errors, we thus obtain an accurate solution for the camera parameters. We also extend our approach to a calibration technique using images of a 1-D object with a fixed pivoting point. Unlike existing methods that rely on orthogonality or pole-polar relationship, our approach utilizes new inter-image constraints and does not require knowledge of the 3-D coordinates of feature points. To demonstrate the effectiveness of the approach, we present results for both synthetic and real images. PMID- 17076421 TI - Accentuate the positive. PMID- 17076422 TI - Transient modeling in simulation of hospital operations for emergency response. AB - Rapid estimates of hospital capacity after an event that may cause a disaster can assist disaster-relief efforts. Due to the dynamics of hospitals, following such an event, it is necessary to accurately model the behavior of the system. A transient modeling approach using simulation and exponential functions is presented, along with its applications in an earthquake situation. The parameters of the exponential model are regressed using outputs from designed simulation experiments. The developed model is capable of representing transient, patient waiting times during a disaster. Most importantly, the modeling approach allows real-time capacity estimation of hospitals of various sizes and capabilities. Further, this research is an analysis of the effects of priority-based routing of patients within the hospital and the effects on patient waiting times determined using various patient mixes. The model guides the patients based on the severity of injuries and queues the patients requiring critical care depending on their remaining survivability time. The model also accounts the impact of prehospital transport time on patient waiting time. PMID- 17076423 TI - Anti-personnel landmine injuries during peace: experience in a European country. AB - INTRODUCTION: The purpose of this study is to report the incidence of landmine injuries during peacetime in a European country. METHODS: Forty victims of landmine explosions were admitted to Didimoticho General Hospital in Greece, from December 1988 to March 2003. A total of 19 people survived (47.5%) these events; all of the others were dead upon admission to the hospital. All of the victims were men, either suspected smugglers or migrants entering the country illegally, with an average of 30 years (range: 15-56 years). RESULTS: Most victims presented in groups, with multiple traumatic injuries, including lower extremity wounds. The mortality rate in the minefield prior to hospital admission was 52.5%, and the amputation rate for the survivors was 37%. There were no deaths of the patients admitted to the hospital. CONCLUSIONS: Landmines cause high-energy injuries with high mortality and amputation rates. Illegal migrants are the main victims of landmine explosions in Greece. PMID- 17076424 TI - Reanimation Unit experience of a second-level hospital in Mexico City. AB - OBJECTIVES: The objectives of this study were to determine the clinical characteristics of patients who presented to the Reanimation Unit (RU) of a second-level hospital during one year, and the number and type of emergency procedures performed. METHODS: A cross-sectional study was designed that enrolled all patients >15 years of age who presented to the RU from 01 January through 31 December 2003. The age, gender, diagnosis, site of origin, and disposition of each patient was recorded, as well as the distribution by time of day, the number and type of emergency procedures performed, complications, and mortality rate. RESULTS: Of the 3,741 patients enrolled in the study, 57.0% were male; predominantly 41-50 years old (20%). Most patients presented to the RU from their homes during the afternoon. There were 60 different admission diagnoses: more of the emergencies were for medical than for traumatic emergencies. The predominant pathologies were bronchospasm, hypertensive crisis, and upper gastrointestinal bleeding. Initially, patients either were admitted to the observation unit, the consulting office for the emergency department, or the intensive care unit. There were a total of 2,753 emergency procedures performed: orotracheal intubations were the most common, followed by installation of a catheter into the central venous circulation. Of all of the patients admitted to the RU, 31% were not insured. CONCLUSIONS: There exists a remarkable combination between medical and traumatic emergencies, which is not encountered frequently in other second level hospitals in Mexico City. A high proportion of the patients who received medical attention were not insured and there were a large number of emergency invasive procedures performed. PMID- 17076425 TI - What a disaster?! Assessing utility of simulated disaster exercise and educational process for improving hospital preparedness. AB - INTRODUCTION: Recent events have brought disaster medicine into the public focus. Both the government and communities expect hospitals to be prepared to cope with all types of emergencies. Disaster simulations are the traditional method of testing hospital disaster plans, but a recent, comprehensive, literature review failed to find any substantial scientific data proving the benefit of these resource- and time-consuming exercises. OBJECTIVES: The objective of this study was to test the hypothesis that an audiovisual presentation of the hospital disaster plans followed by a simulated disaster exercise and debriefing improved staff knowledge, confidence, and hospital preparedness for disasters. METHODS: A survey of 50 members of the medical, nursing, and administrative staff were chosen from a pool of approximately 170 people likely to be in a position of responsibility in the event of a disaster. The pre-intervention survey tested factual knowledge as well as perceptions about individual and departmental preparedness. Post-intervention, the same 50 staff members were asked to repeat the survey, which included additional questions establishing their involvement in the exercise. RESULTS: There were 50 pre-intervention tests and 42 post intervention tests. The intervention resulted in a significant improvement in test pass rate: pre-intervention pass rate 9/50 (18%, 95% confidence interval ((CI) = 16.1-19.9%) versus post-intervention pass rate 21/42 (50%, 95% CI = 42.4 57.6%; chi-square test, p = 0.002). Emergency department (ED) staff had a stronger baseline knowledge than non-ED staff: ED pre-test mean value for scores = 12.1 versus non-ED scores of 6.2 (difference 5.9, 95% CI = 3.3-8.4); t-test, p <0.001. Those that attended > or = 1 component had a greater increase in mean scores: increase in mean attendees was 5.6, versus the scores of non-attendees of 2.7 (difference 2.9, 95% CI = 1.0-4.9); t-test, p = 0.004. There was no significant increase in the general perception of preparedness. However, the majority of those surveyed described the exercise of benefit to themselves (53.7%, 95% CI = 45.5-61.8%) and their department (63.2%, 95% CI = 53.5-72.8%). CONCLUSIONS: The disaster exercise and educational process had the greatest benefit for individuals and departments involved directly. The intervention also prompted enterprise-wide review, and an upgrade of disaster plans at departmental levels. Pre-intervention knowledge scores were poor. Post-intervention knowledge base remained suboptimal, despite a statistically significant improvement. This study supports the widely held belief that disaster simulation is a worthwhile exercise, but more must be done. More time and resources must be dedicated to the increasingly important field of hospital disaster preparedness. PMID- 17076426 TI - Weapons of mass destruction preparedness and response for the XIV Pan-American Games, Santo Domingo, 2003. AB - The Pan-American Games are considered to be the fourth most important international athletic event in the world. Hosted by the city of Santo Domingo, Dominican Republic, the XIV Pan-American Games lasted from 11-17 August 2003. In preparation for the Games, the Security Directorate developed and deployed a Weapons of Mass Destruction (WMD) Unit. For operational support, two strike teams (Alpha and Bravo) were active at any given time. Each team consisted of five members including a team leader, field physician, explosive ordinance disposal (EOD) officer, and two tactical officers. Three hospitals-two military and one civilian--were designated as primary medical centers for the event. With the assistance of the WMD Unit, emergency department staff were trained in the medical management of a WMD event, response protocols for WMD were created, and special decontamination areas were designated. Syndromic surveillance was performed by means of direct communications between the hospitals and units, as well as use of an electronic, Web-based surveillance tool. PMID- 17076427 TI - Paramedic student performance: comparison of online with on-campus lecture delivery methods. AB - INTRODUCTION: Colleges and universities are experiencing increasing demand for online courses in many healthcare disciplines, including emergency medical services (EMS). Development and implementation of online paramedic courses with the quality of education experienced in the traditional classroom setting is essential in order to maintain the integrity of the educational process. Currently, there is conflicting evidence of whether a significant difference exists in student performance between online and traditional nursing and allied health courses. However, there are no published investigations of the effectiveness of online learning by paramedic students. HYPOTHESIS: Performance of paramedic students enrolled in an online, undergraduate, research methods course is equivalent to the performance of students enrolled in the same course provided in a traditional, classroom environment. METHODS: Academic performance, learning styles, and course satisfaction surveys were compared between two groups of students. The course content was identical for both courses and taught by the same instructor during the same semester. The primary difference between the traditional course and the online course was the method of lecture delivery. Lectures for the on-campus students were provided live in a traditional classroom setting using PowerPoint slides. Lectures for the online students were provided using the same PowerPoint slides with prerecorded streaming audio and video. RESULTS: A convenience sample of 23 online and 10 traditional students participated in this study. With the exception of two learning domains, the two groups of students exhibited similar learning styles as assessed using the Grasha Riechmann Student Learning Style Scales instrument. The online students scored significantly lower in the competitive and dependent dimensions than did the on campus students. Academic performance was similar between the two groups. The online students devoted slightly more time to the course than did the campus students, although this difference did not reach statistical significance. In general, the online students believed the online audio lectures were more effective than the traditional live lectures. CONCLUSION: Distance learning technology appears to be an effective mechanism for extending didactic paramedic education off-campus, and may be beneficial particularly to areas that lack paramedic training programs or adequate numbers of qualified instructors. PMID- 17076428 TI - Medical preparedness against chemical and biological incidents for the NATO Summit in Istanbul and lessons learned. AB - INTRODUCTION: During the 2004 North Atlantic Treaty Organization (NATO) Summit, essential counter-measures, including medical preparedness, were taken to cope with any suspected terrorist case or events including the use of chemical or biological (CB) weapons. The Summit was held in Istanbul, a city that bridges two continents, and involved the participation of many Heads of State, Prime Ministers, and Defense Ministers from 26 NATO countries. METHODS: First responders, including medical Chemical, Biological, Radiological, and Nuclear (CBRN) teams, received special training. Essential equipment, including drugs, antidotes, detectors, etc., was provided and stock-piled. Medical authorities augmented the capacity for identifying and controlling the injuries and any emerging CB incident through the set-up of decontamination units and the procurement of medical devices, antidotes, drugs, and personal protective suits. Additionally, a small part of the recently established NATO-CBRN battalion was welcomed to the Summit and was prepared to perform detection and identification of the agent found in suspicious appearing samples. RESULTS: Although no CB incident was reported during the Summit, extensive experience was gained with respect to medical preparedness against CB terrorism. Sampling, detection, and analysis of toxic materials were taken into account in the medical management. Much laboratory-related work was conducted in the following time period. The laboratory work involved the standardization of sampling and transportation procedures, development of both mobile and reference laboratories, and performing research activities aimed to make the CB analysis more efficient. Although the training of the medical staff was advanced, training should be continuous and supported with educational programs, conferences, meetings, and tabletop and hospital medical exercises throughout the country. CONCLUSION: Multidisciplinary cooperation, training, and preparedness should be provided to basic medical care units and centers as part of the medical planning aimed at perfect detection and surveillance, laboratory analysis, and emergency response. PMID- 17076429 TI - Using innovative simulation modalities for civilian-based, chemical, biological, radiological, nuclear, and explosive training in the acute management of terrorist victims: A pilot study. AB - OBJECTIVES: Chemical, biological, radiological, nuclear, and explosive (CBRNE) incidents are low frequency, high impact events that require specialized training outside of usual clinical practice. Educational modalities must recreate these clinical scenarios in order to provide realistic first responder/receiver training. METHODS: High fidelity, mannequin-based (HFMB) simulation and video clinical vignettes were used to create a simulation-based CBRNE course directed at the recognition, triage, and resuscitation of contaminated victims. The course participants, who consisted of first responders and receivers, were evaluated using a 43-question pre- and post-test that employed 12 video clinical vignettes as scenarios for the test questions. The results of the pre-test were analyzed according to the various medical training backgrounds of the participants to identify differences in baseline performance. A Scheffe post-hoc test and an ANOVA were used to determine differences between the medical training backgrounds of the participants. For those participants who completed both the pre-course and post-course test, the results were compared using a paired Student's t-test. RESULTS: A total of 54 first responders/receivers including physicians, nurses, and paramedics completed the course. Pre-course and post-course test results are listed by learner category. For all participants who took the pre-course test (n = 67), the mean value of the test scores was 53.5 +/- 12.7%. For all participants who took the post-course test (n = 54), the mean value of the test scores was 78.3 +/-10.9%. The change in score for those who took both the pre- and post-test (n = 54) achieved statistical significance at all levels of learner. CONCLUSIONS: The results suggest that video clinical vignettes and HFMB simulation are effective methods of CBRNE training and evaluation. Future studies should be conducted to determine the educational and cost-effectiveness of the use of these modalities. PMID- 17076430 TI - Prevalence of radioactive signals from surveillance of an emergency department. AB - INTRODUCTION: Since the 11 September 2001 terrorist attacks in the United States, concerns have been raised regarding the threat of a radiological terrorist weapon. Although the probability of the employment of a nuclear device is remote, the potential of a radiological dispersal device (RDD) or "dirty bomb" is of concern. While it is unlikely that such a device would produce massive numbers of casualties, it is far more likely that it would result in public panic and perhaps even disable the local healthcare system. The utility of surveillance with radiation detectors in the healthcare setting has not been fully evaluated. OBJECTIVE: The objective of this study was to characterize the prevalence of radioactive sources entering an urban emergency department (ED). METHODS: A retrospective review of data obtained from a radiation detector positioned to detect radioactive people entering an ED of an urban academic hospital that serves 45,000 patients/year was performed. Graphical outputs of radioactivity were recorded in Microsoft Excel (Microsoft, Redmond, WA, US) spreadsheets in microREM/hour. Data were collected continuously from 22 December 2003 to 22 January 2004. An event was defined as any elevation in radiation levels >95% confidence interval from the mean level of background radiation over 72 hours (h). RESULTS: A total of 215 events were observed over a 28-day period, with a mean value of 7.7 events/day, and a maximum of 15 events/day. During the 28-day period, the baseline mean level of background radiation was 2-4 microREM/h. Readings ranged from 2,148.28-17,292.25 microREM/h with a maximum sustained detector exposure of 684.37 microREM. Distinct signal patterns were seen at both detectors including tonic, phasic, dual, and short duration spikes. CONCLUSION: The number of radioactive signals detected from persons entering the ED was much higher than expected. While the vast majority of these signals pose no health threat, they may make routine screening for a radiological terrorist event difficult. Further study is needed to determine this correlation. PMID- 17076431 TI - Medical support during the European Union Summit in Gothenburg, Sweden, June 2001. AB - INTRODUCTION: Several factors are important for the number and severity of medical emergencies during mass-gatherings. The risk of violence, the size and mobility of the crowd, the type of event, weather, and duration of the event all influence the outcome. During the European Union (EU) Summit, from 15-16 June 2001 in Gothenburg, Sweden, approximately 50,000 people participated in 43 protest marches, some which included 15,000 participants. Clashes between police and the protesters occurred. OBJECTIVE: The objective of this study was to analyze the amount and character of injuries as well as the medical complaints in relation to the EU Summit. In addition, the aim of this study was to describe the organization and function of the healthcare services provided during the meeting. METHODS: This study is based on the medical records of patients presenting with injuries and other types of medical emergencies at the healthcare stations during the Summit. RESULTS: In total, 143 patients sought medical care. Fifty-three (37.1%) were police officers. Most patients had minor complaints, but a few were seriously injured. The Patient Presentation Rate (PPR) was 2.7. Nine victims were hospitalized as high priority. CONCLUSION: The PPR for the EU Summit was 2.7, which is in the same range as previously reported from other mass-gatherings. PMID- 17076432 TI - Difficult discussions: military intervention and United Nation reform. PMID- 17076433 TI - Parameterizing the attentional blink effect. AB - The attentional blink effect (AB) is used to examine the limits of attention in dual-task paradigms. However, since the effect is nonlinear, it is sometimes difficult to characterize the results. Furthermore, it is difficult to assess the significance of the effect between groups because the results are highly variable both within and across subjects. In this paper, we propose a method to quantify four characteristics of the AB curve: the minimum performance, the amplitude between the minimum and the asymptotic performance, the amount of Lag-1 sparing, and the width of the effect. The method, based on curve fitting, allows easier comparisons of the results across experiments, can test only one characteristic at a time, and yields more powerful statistical tests. PMID- 17076434 TI - Dissociative effects of stimulus quality on semantic and morphological contexts in visual word recognition. AB - Semantic and morphological contexts were manipulated jointly with stimulus quality under conditions where there were few related prime-target pairs (i.e., low relatedness proportion) in a lexical decision experiment. Additive effects of semantic context and stimulus quality on RT were observed, replicating previous work. In contrast, morphological context interacted with stimulus quality. This dissociation is discussed in the context of Besner and colleagues' evolving multistage framework. The essence of the account is that 1) stimulus quality affects feature and letter levels, but not later levels, 2) feedback from semantics to the lexical level is inoperative under low relatedness proportion conditions (hence stimulus quality and semantic context yield additive effects), whereas 3) feedback from the lexical level to the letter level is intact, hence stimulus quality and morphological context produce an interaction by virtue of them affecting a common stage of processing. PMID- 17076435 TI - The influence of advance information about target location and visual feedback on movement planning and execution. AB - This study was designed to determine if movement planning strategies incorporating the use of visual feedback during manual aiming are specific to individual movements. Advance information about target location and visual context was manipulated using precues. Participants exhibited a shorter reaction time and a longer movement time when they were certain of the target location and that vision would be available. The longer movement time was associated with greater time after peak velocity. Under conditions of uncertainty, participants prepared for the worst-case scenario. That is, they spent more time organizing their movements and produced trajectories that would be expected from greater open-loop control. Our results are consistent with hierarchical movement planning in which knowledge of the movement goal is an essential ingredient of visual feedback utilization. PMID- 17076436 TI - Selection of procedures in mental subtraction. AB - Adults solved simple subtraction problems (e.g., 16 - 9). Half of the 32 participants provided immediate self-reports of their solution processes on each problem. Performance was analyzed using both traditional descriptive statistics (i.e., means, standard deviations, and percentage of errors) and with statistics derived from fitting the ex-Gaussian distributions to latencies (i.e., mu and tau). The results support the view that ex-Gaussian analyses can be useful in exploring patterns of procedure selection that relate both to characteristics of the stimuli (e.g., problem size) and to characteristics of the participants (e.g., arithmetic skill). More generally, the results provide further evidence that adults use a variety of procedures to solve simple subtraction problems and that these choices are related to patterns of performance on more complex problems that require calculation. PMID- 17076437 TI - [French norms of imagery for pictures, for concrete and abstract words]. AB - This paper deals with French norms for mental image versus picture agreement for 138 pictures and the imagery value for 138 concrete words and 69 abstract words. The pictures were selected from Snodgrass et Vanderwart's norms (1980). The concrete words correspond to the dominant naming response to the pictorial stimuli. The abstract words were taken from verbal associative norms published by Ferrand (2001). The norms were established according to two variables: 1) mental image vs. picture agreement, and 2) imagery value of words. Three other variables were controlled: 1) picture naming agreement; 2) familiarity of objects referred to in the pictures and the concrete words, and 3) subjective verbal frequency of words. The originality of this work is to provide French imagery norms for the three kinds of stimuli usually compared in research on dual coding. Moreover, these studies focus on figurative and verbal stimuli variations in visual imagery processes. PMID- 17076438 TI - Social and hormonal bases of individual differences in the parental behaviour of birds and mammals. AB - We present an overview of research on how social experiences and hormonal responses affect individual variation in parental care of birds and mammals. The parental roles of prolactin and glucorticoids (corticosterone or cortisol) have many similarities in birds and in mammals. Prolactin may be involved in the initiation of parental interactions, with prolactin variation possibly explaining individual differences in parental decision-making. Glucocorticoid levels increase when parents have to work harder, with some individuals showing greater hormonal and behavioural responses than others. Testosterone interferes with paternal behaviour in birds, but its role is more complex and species-specific in male mammals. We examine these differences in an adaptive framework, where retaining flexibility of response has allowed individuals to respond differentially to social opportunities and environmental change. PMID- 17076439 TI - The military physician as a noncombatant: the concept of over identification. PMID- 17076440 TI - Imported malaria and conflict: 50 years of experience in the U.S. Military. AB - Over the last 50 years, members of the U.S. military have faced the threat of malaria in diverse geographical locations and operational situations, resulting in considerable morbidity and mortality. However, because malaria may be transported out of endemic areas and into areas that are free of malaria, the threat does not end with redeployment. Since the Korean Conflict, outbreaks of imported malaria have followed every major deployment of U.S. military forces to malaria endemic areas. By examining unique aspects of these outbreaks through the years, many similarities can be drawn. Repeated observations demonstrate that preventive efforts are only effective at reducing the impact of imported malaria when commanders are informed about the risks of malaria. Commanders must also understand the preventive resources that are available while deployed, and the potential for morbidity and mortality from malaria. PMID- 17076441 TI - Summary of the 2004-2005 influenza season in the U.S. Army Europe. AB - Influenza and other respiratory infections, the most common cause of acute infectious disease in U.S adults, are also the leading cause of outpatient illness and a major cause of infectious disease hospitalization in U.S. military personnel. Although respiratory disease control is improved, epidemics continue to occur, and respiratory disease in military trainees continues to exceed that in U.S. civilian adults. Overall, Department of Defense utilization of the trivalent inactivated vaccine was much lower than anticipated during the 2004 2005 season. The slow start to the 2004-2005 influenza season resulted in a low demand for influenza immunization by the medically high-risk beneficiary population of the Department of Defense. Surveillance for influenza during the 2004-2005 season in U.S. Army Europe reached unprecedented heights, testing and confirming more cases than in any previous year. PMID- 17076442 TI - Development of a deployment course for graduating military internal medicine residents. AB - Graduates of military internal medicine residency programs are required to have the necessary knowledge and skills to function as internists, military physicians, and military medical leaders. The global war on terrorism has increased the role internists are playing in combat theaters as they fill multiple different military medical positions including battalion, brigade, and division surgeons as well as physicians in echelon I, II, and III medical facilities. Along with general internists, internal medicine subspecialists, pediatricians, and family physicians also fill these roles. Although internal medicine training provides a broad-based knowledge to care for adults, it does not provide significant training in combat casualty care, detainee health care, or environmental health. To overcome many of these perceived shortfalls, we developed the 3-day deployment course for graduating internal medicine residents outlined in this article. Through a combination of didactic and hands-on training, militarily relevant medical knowledge and skills necessary to function at echelon I and II levels of care were provided. Residents uniformly accepted the course with measurable increase in their fund of knowledge at the completion of the course. PMID- 17076443 TI - Comparing the population health impacts of medical conditions using routinely collected health care utilization data: nature and sources of variability. AB - Prevention activities are designed and resourced based on perceptions of the relative population health impacts of various conditions. We examined the nature and variability of rankings of "conditions" based on how they are defined and how their population health impacts are measured. The first listed diagnosis from all hospitalizations and ambulatory visits of U.S. service members during 2002 was used to rank conditions (as defined by two standard classification systems) using five different measures of population health impacts. Less than 10% of all conditions accounted for more than one-half of total population health impact, regardless of how conditions were defined or impacts measured. However, specific conditions with the largest impacts varied depending on the classification system and impact measure. Four groups of related conditions--acute musculoskeletal injuries, pregnancy-related conditions, respiratory infections, and mental disorders (including substance abuse)--accounted for disproportionately large impacts regardless of the measure. The identification of conditions with the largest population health impacts depends on the nature and degree of aggregation in defining conditions and the measure of impact. The findings are relevant to prevention planning and resourcing. PMID- 17076444 TI - Veterans and functional status transitions in older Americans. AB - OBJECTIVES: This research examines the relationship of veteran status with functional status transitions in older Americans. METHODS: Data for this study come from the Survey of Asset and Health Dynamics among the Oldest Old. We use a structural multinomial logit model to decompose the effect of older veterans into the direct effect and the indirect effects via physical health conditions and mental disorders on functional status transitions. RESULTS: Although there is no distinct association among those functionally independent at baseline, veteran status significantly impacts age-dependent transitions from functional dependence to other statuses. At age 85, the excess mortality and the lower level of functional resolution among functionally dependent veterans are considerable. CONCLUSIONS: Physical health is more important than mental health in transmitting the effect of veteran status on functional status transitions in functionally dependent persons. PMID- 17076445 TI - Differences in beneficiary assessments of health care between TRICARE prime and TRICARE prime remote. AB - OBJECTIVES: We examined differences in health care ratings and reported health care experiences for active duty uniform services personnel using health care plans other than military treatment facilities. METHODS: We used a cross sectional mail survey of a stratified sample of 3,871 beneficiaries enrolled in TRICARE Prime (TP) and TRICARE Prime Remote (TPR). The adjusted plan mean composite and global ratings were compared between TP and TPR participants. RESULTS: There were few significant differences between the two groups. Patient satisfaction was higher when patients chose their providers (TPR), and use of some preventive services was higher in managed-care plans (TP). Respondents in metropolitan locations differed significantly from those in nonmetropolitan locations in ratings of plans, quality of health care received, and access to services. CONCLUSIONS: The military health system is achieving some success in delivering uniform benefits but faces challenges in delivering high-quality uniform benefits in rural communities. PMID- 17076446 TI - Canadian forces evaluation of the EPINATO Health Surveillance System in Bosnia Herzegovina. AB - The Canadian Forces (CF) adopted the EPINATO surveillance system in 1996 to monitor disease and injury morbidity in deployed settings. The Directorate of Force Health Protection, CF Health Services Group initiated an evaluation of EPINATO in Task Force Bosnia-Herzegovina in August 2003. Two methods were used to assess coding reliability: a chart audit and Sick Parade Register review. Stakeholder interviews were conducted evaluating data flow, reporting structure, and key system attributes. Reliability (K, 95% confidence interval) was good in 4 of 24 categories--sexually transmitted diseases, K = 0.75 (0.50, 1.00); eye disorders, K = 0.51 (0.15, 0.88); ears/nose/ throat, K = 0.51 (0.33, 0.69); lower respiratory infections, K = 0.49 (95% confidence interval 0.34, 0.65)-but otherwise was poor. EPINATO is not an effective, reliable tool for CF deployment health surveillance. An improved health surveillance system is required to ensure disease and injury aberrations are detected and optimal preventive programs and policies are in place for deployed CF military members. PMID- 17076447 TI - Modeling an integrative physical examination program for the Departments of Defense and Veterans Affairs. AB - Current policies governing the Departments of Defense and Veterans Affairs physical examination programs are out of step with current evidence-based medical practice. Replacing periodic and other routine physical examination types with annual preventive health assessments would afford our service members additional health benefit at reduced cost. Additionally, the Departments of Defense and Veterans Affairs repeat the physical examination process at separation and have been unable to reconcile their respective disability evaluation systems to reduce duplication and waste. A clear, coherent, and coordinated strategy to improve the relevance and utility of our physical examination programs is long overdue. This article discusses existing physical examination programs and proposes a model for a new integrative physical examination program based on need, science, and common sense. PMID- 17076448 TI - Comparison of nondeployable hearing profiles by Army component (Active Duty, National Guard, and Reserve) and by gender. AB - The Hearing Conservation Service at Fort Bliss conducted baseline-hearing assessments from January 2003 to March 2005 on 14,974 soldiers in the Active Duty (AD), National Guard (NG), and Reserve Component (RC). The percentages of soldiers with >H-2 (i.e., nondeployable) hearing profiles based on Army component and gender are as follows: AD male = 2.21%; AD female = 0.65%; NG male = 5.05%; NG female = 1.64%; RC male = 5.02%; and RC female = 0.54%. Female soldiers in all three components and AD male soldiers are not significantly different (p < 0.05) from each other and have a significantly lower percentage of >H-2 hearing profiles than NG and RC male soldiers after adjusting for age. With the Army engaged in worldwide missions, these results should prove useful to commanders and medical officers for assessing deployment capabilities and for hearing conservation interventions. PMID- 17076449 TI - The impact of a situationally focused individual human immunodeficiency virus/sexually transmitted disease risk-reduction intervention on risk behavior in a 1-year cohort of Nigerian military personnel. AB - Human immunodeficiency virus (HIV) and sexually transmitted disease (STD)-related risks in peacekeeping troops is a concern when they are stationed in areas of high HIV prevalence. We carried out an assessment of a situationally focused individual HIV/STD reduction intervention (where avoidance of risk situations, as well as risk behaviors, are emphasized) in one Nigerian military unit (N = 1,222), with a comparable unit from the same service as a waiting list control (N = 987). The intervention consisted of a possible five modules that were presented to groups of up to 50 personnel. Data were collected on reported sexual behaviors, condom beliefs, sexual risk behaviors with casual partners, and number of interventions attended. Data indicated significant increases in reported condom use with casual partners and positive condom beliefs at 6- and 12-month follow-up. Risk behavior was reduced 30% from baseline at 6 months and 23% from baseline at 12 months. There was also a significant dose-response effect for number of interventions attended. These data suggest that relatively brief situationally focused individual interventions are effective in military and West African contexts in reducing HIV/STD risk behaviors. PMID- 17076450 TI - Auditory perception with ear and cold weather face protection worn in combination. AB - The effects on hearing thresholds, sound attenuation, and consonant discrimination of wearing a balaclava under hearing-protecting earmuffs were studied. This combination is commonly worn during cold weather military operations. One group of 20 normal-hearing adults (10 male and 10 female subjects) was tested. Within-subject measurements were made of diffuse-field hearing thresholds from 0.25 kHz to 8 kHz and consonant discrimination in quiet with the ears unoccluded and protected with the earmuffs alone and with the balaclava worn full face or rolled. Attenuation was derived from the protected and unoccluded thresholds at each frequency. When the balaclava was worn full face, attenuation decreased by 16 to 18 dB, relative to the muff alone, below 6.3 kHz. With the balaclava worn as a cap, there was an inverted U-shaped decrement in attenuation of 18 to 27 dB from 0.25 Hz to 4 kHz. Consonant discrimination decreased by 7% with the muffs alone. These findings underscore the importance of assessing protective equipment under the conditions in which it will be worn. PMID- 17076451 TI - Reference values for performance on the Automated Neuropsychological Assessment Metrics V3.0 in an active duty military sample. AB - The Automated Neuropsychological Assessment Metrics (ANAM) is a computerized measure of processing speed, cognitive efficiency, and memory. This study describes performance and psychometric properties of ANAM in an active duty, healthy military sample (N = 2,371) composed primarily of young (18-46 years) adult males. Rarely have neuropsychological reference values for use with individuals in the military been derived from a large, active duty military population, and this is the first computerized neuropsychological test battery with military-specific reference values. Although these results do not provide demographically corrected, formal normative data, they provide reference points for neuropsychologists and other health care providers who are using ANAM data in research or clinical settings, with patients of comparable demographics to the present sample. PMID- 17076452 TI - Recurrent idiopathic nonimmune hydrops fetalis: A case report. AB - A recurrent idiopathic nonimmune hydrops fetalis case in two subsequent pregnancies was observed in a woman with no child. In both pregnancies, all the detailed analysis, including a high level of ultrasonography, amniocentesis, serologic evaluation, routine blood work, hemoglobin electrophoresis, and glucose 6-phosphate/dehydrogenase level and postdelivery autopsy, was done, and after no clue of etiologic agent, pregnancy termination was applied for both. This case report illustrates the importance of recurrence of nonimmune hydrops fetalis in the absence of any etiology. Mostly, it is hard to establish an etiologic diagnosis for adequate management of subsequent pregnancies. PMID- 17076453 TI - Usefulness of temazepam and zaleplon to induce afternoon sleep. AB - Insufficient daytime sleep may result in reduction of effectiveness and safety during overnight military missions. The usefulness of temazepam and zaleplon to optimize afternoon sleep and their effects on performance and alertness during a subsequent night shift were studied. METHOD: In a randomized double-blind within subjects design, 11 subjects took 20 mg of temazepam, 10 mg of zaleplon, or placebo before a 5:30-10:00 p.m. sleep period. Sleep length and quality were measured. Subjects were kept awake throughout the night while alertness, cognitive performance, and muscle power were repeatedly measured. RESULTS: Temazepam provided significantly longer and qualitatively better sleep than zaleplon or placebo. During the night, sleepiness increased and muscle power was impaired in all conditions. Better sleep was correlated with less sleepiness during the night. CONCLUSION: Temazepam is useful to optimize a 4.5-hour afternoon sleep before overnight missions. Irrespective of hypnotic treatment, sleepiness and fatigue increased during the night shift. PMID- 17076454 TI - Challenges to military working dog management and care in the Kuwait theater of operation. AB - In a combat environment, troop safety and installation security are paramount. Documentation of the frequent use of explosive devices by insurgents in Southwest Asia has increased the need for augmentation of defensive capabilities. Military working dogs (MWDs) are trained and certified to perform missions supporting security and detection of explosives. Challenges arise in theater because the number of certified dogs available can be limited and the standards of housing and care that are available in the continental United States are not usually available in a combat theater. Planning, preparation, and training of MWD handlers,animal care specialists, and Veterinary Corps officers will maximize mission performance and MWD health. Some of th echallenges and needs associated with management and care of the MWDs in the Kuwait theater of operation are discussed. Suggested improvements to the current deployment readiness paradigm are offered from the perspective of a staff veterinarian who was deployed in support of Operation Iraqi Freedom in 2004. PMID- 17076455 TI - Influence of oral hygiene on oral health of recruits and professionals in the Croatian Army. AB - OBJECTIVES: Good oral health of soldiers would decrease the number of urgent dental interventions and absences from training and the battlefield and would improve the security of the whole formation. This study shows the habits for maintaining oral health and the oral status of the examined population in the Croatian Army. METHODS: The data were obtained from examinations and questionnaires of 912 Croatian soldiers, 650 of whom were recruits and 262 professional soldiers of the Croatian Army land forces. RESULTS: The results showed that the oral health of the examined population was mostly bad, as a consequence of inadequate prevention of illnesses of the oral cavity associated with insufficient oral hygiene. The most common disease was dental caries (5.84 carious teeth per recruit and 2.71 per professional). Only 14 (1.53%) of 912 examinees had completely healthy teeth. Only one-third of the examinees had no bleeding when tested with a periodontal probe. Acute pain of odontogenic origin was present for 23.5% of examinees. Such oral health makes most of the soldiers unreliable for peace operations that would last >6 months, because it lowers their combat readiness. CONCLUSION: Oral hygiene and oral health are poor. Consequently, combat readiness is low because of the need for frequent dental interventions, which could further lead to absence from the field and appointed tasks. The results emphasize the need for obligatory regular check-ups to improve oral health in the Croatian Army. PMID- 17076456 TI - Auricular acupuncture in the treatment of acute pain syndromes: A pilot study. AB - This pilot study used a randomized controlled clinical trial design to compare the effects of standard emergency medical care to auricular acupuncture plus standard emergency medical care in patients with acute pain syndromes. Eighty seven active duty military personnel and their dependents with a diagnosis of acute pain completed the study, which was conducted in the emergency room (ER) at Malcolm Grow Medical Center, Andrews Air Force Base, Maryland. The primary outcome measure was change in pain level from baseline, as measured by the Numerical Rating Scale. Participants in the acupuncture group experienced a 23% reduction in pain before leaving the ER, while average pain levels in participants in the standard medical care group remained basically unchanged. (p < 0.0005). However, both groups experienced a similar reduction in pain 24 hours following treatment in the ER. More research is needed to elucidate treatment effects and to determine mechanisms. PMID- 17076457 TI - Factors associated with vaginal douching in military women. AB - Objectives of this secondary analysis were to describe the prevalence of vaginal douching among 1,432 women on active duty in the military and to examine demographic and behavioral factors associated with douching. More than one-half (54.5%) of the respondents reported douching at least once in their lifetimes; 63.5% of those douched during the previous year, and 45.8% of those douched in > or = 6 of the past 12 months. Multiple regression analysis showed that being African American, using tampons, having sexual intercourse, having more than one sexual partner in a month, using spermicides, and being in the enlisted and noncommissioned officer ranks were associated with more frequent douching (20.1% of the variance in douching behavior was explained). Despite access to regular gynecological examinations and related health counseling, a significant number of military women continue to douche. These women may benefit from specific intensive information on normal vaginal self-cleansing mechanisms and the health hazards of douching. Military efforts to diminish this self-care behavior and to enhance providers' knowledge about the scope of the problem may improve women's health and military readiness by decreasing the incidence of negative consequences of douching. PMID- 17076458 TI - A cross-sectional survey of stressors for postpartum women during wartime in a military medical facility. AB - OBJECTIVE: To determine whether having a partner deployed during wartime increased the stress levels in pregnant women and altered the attitudes toward pregnancy or changed birth outcomes. METHODS: Cross-sectional survey of all postpartum women at Naval Hospital Camp Lejeune. The anonymous survey was administered from May to July 2003. RESULTS: Ninety-five surveys were collected. Fewer patients reported that their partner was deployed (41.1%) than not deployed (58.9%). Women with deployed partners gave birth to larger babies (3526.5 g vs. 3248.7 g, p = 0.016). No difference was seen in the gestational age at delivery, percentage with vaginal delivery, average number of children at home, self reported stress, or reported weight gain during pregnancy. Women with partners deployed more often reported changed eating habits (56.4% vs. 8.0%, p < 0.001). Those with a deployed partner more often reported that media coverage impacted their stress level (p = 0.003). CONCLUSIONS: Pregnant women with deployed partners gave birth to larger babies. They also more frequently report a change in eating habits and that media coverage impacted their stress level. PMID- 17076459 TI - Sexually transmitted diseases in Operation Iraqi Freedom/Operation Enduring Freedom. AB - OBJECTIVE: To identify the incidence of sexually transmitted diseases (STDs) in a female active duty population deployed in support of Operation Iraqi Freedom/Operation Enduring Freedom was the objective of this study. METHODS: Retrospective chart review was completed on all soldiers seeking outpatient gynecologic care at Camp Doha, Kuwait, from September 2003 through March 2004. Descriptive statistical analysis was performed on data from all patients identified as having an STD. RESULTS: Forty-four soldiers (2.5% of all encounters) were diagnosed with STDs during the study period. Genital herpes, Condyloma acuminata, and chlamydia were the most commonly identified infections accounting for 30, 25, and 21% of the diagnoses, respectively. CONCLUSION: Transmission of STDs in the deployed environment continues to be problematic. Viral infections were more commonly encountered than were bacterial infections. Patient education and prevention should be emphasized. Consideration should be given to STD screening upon redeployment. PMID- 17076461 TI - "Uncoverage" of the humeral head by the anterolateral acromion and its relationship to full-thickness rotator cuff tears. AB - The relationship between the humeral head and the anterolateral acromion was compared for subjects with and without full-thickness rotator cuff tears, as indicated by magnetic resonance imaging, using a case-control design. "Uncoverage" of the humeral head by the anterolateral acromion was defined as the percentage of the humeral head width not covered superiorly by the anterolateral acromion, as measured digitally on magnetic resonance imaging scans of the shoulder in the oblique coronal plane. The uncoverage of the humeral head by the anterolateral acromion was 32.0% for men > or = 40 years of age with normal rotator cuffs (n = 39) and 38.2% for men > or = 40 years of age with full thickness rotator cuff tears (n = 43). The difference in uncoverage of 6.2% was statistically significant (p < 0.001). The uncoverage of the humeral head by the anterolateral acromion was 36.1% for women > or = 40 years of age with normal rotator cuffs (n = 10) and 41.8% for women > or = 40 years of age with full thickness rotator cuff tears (n = 17). The difference in uncoverage of 5.72% was not statistically significant with the number of subjects available (p = 0.108). We concluded that increased uncoverage of the humeral head by the anterolateral acromion may be a factor in the pathogenesis of full-thickness rotator cuff tears. PMID- 17076460 TI - Emerging sociodemographic and lifestyle predictors of intention to use condom in human immunodeficiency virus intervention among uniformed services personnel. AB - Appropriate and consistent use of condom remains an effective approach to human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) intervention. We analyzed the baseline data gathered for a situationally based HIV/AIDS intervention to assess the potential predictors of condom use among the Uniformed Services Personnel in Nigeria. Using condom purchase as a proxy for intention to use condom, we examined the distribution of the demographic and lifestyle characteristics, knowledge of HIV transmission mode, and knowledge of how to correctly use a condom. A univariable logistic regression was used to identify the potential predictors, followed by multivariable logistic regression modeling. The knowledge of how to correctly wear a condom was the most significant positive predictor of the intention to use a condom (adjusted prevalence odds ratio (APOR), 5.99 (95% confidence interval (CI), 1.26, 19.79). The other main positive predictors of intent to use a condom were the knowledge of the mode of HIV transmission via blood, APOR 2.43 (95% CI, 1.01, 5.82), saliva (5. 87, 95% CI, 3.15, 10.94), and pre-ejaculatory fluid (APOR, 3.58, 95% CI, 1.67, 7.48). Male gender was also a significant positive predictor of the intent to use a condom (APOR, 2.55, (95% CI, 1.10, 5.97)). The results further indicated alcohol use (APOR, 0.32, 95%CI, 0.16, 0.61), marijuana use (APOR, 0.24, 95% CI, 0.11, 0.56), and the frequency of oral sexual behavior (APOR, 0.006, 95%CI, 0.002, 0.019) as negative predictors of the intent to use a condom. Therefore, these findings suggest that for an HIV/AIDS intervention to be effective in this population, it must incorporate these predictor variables into its design and conduct. PMID- 17076462 TI - Answer to last month's radiology case and images (case # 5): The superior labrum, anterior-to-posterior 'SLAP' lesion. PMID- 17076463 TI - Equine botulism and acute pasture myodystrophy: new soil-borne emerging diseases in Switzerland? AB - In Switzerland, the incidence of equine botulism and acute pasture myodystrophy have remarkably increased in the last five years. Equine fodder-borne botulism in Europe is most likely caused by Clostridium botulinum types C and D that produce the toxins BoNT/C and BoNT/D. Horses showing signs suggestive of botulism (muscle weakness and tremors, reduced tongue tone, slow chewing, salivation and difficulties swallowing, drooping eyelids, mydriasis), especially patients that have fed on suspect fodder (mostly haylage), must be treated with anti-serum as soon as possible. They also need intensive care, which is often difficult to provide and always expensive in the face of a guarded to poor prognosis. Therefore, prevention (high standards of forage quality and vaccination) is all the more important. Pasture myodystrophy is an acute disease with signs of rhabdomyolysis and lethality rate over 90%. It affects grazing horses under frosty, windy and rainy conditions. Preliminary results indicate that Clostridium sordellii and Clostridium bifermentans producing lethal toxin may play a role in pasture myodystrophy. Our efforts concentrate on developing a new subunit vaccine for equine botulism and understanding the ethiology and pathogenesis of pasture myodystrophy with the goal of improving prevention against these highly fatal diseases that present a significant risk to our horse population. PMID- 17076464 TI - Treatment of chronic proximal suspensory desmitis in horses using focused electrohydraulic shockwave therapy. AB - The objective of the present clinical report was to investigate the short- and long-term outcomes of chronic proximal suspensory desmitis (PSD) treated with Extracorporeal Shockwave Treatment (ESWT). Fifty-two horses with chronic PSD in the forelimb (34 cases) or hindlimb (22 cases) were included in the study. Three horses had lesions in both hindlimbs and one in both forelimbs. The origin of the suspensory ligament was treated every three weeks for a total of three treatments using 2000 impulses applied by a focused ESWT device (Equitron) at an energy flux density of 0.15 mJ/ mm2. This treatment regime was followed by box rest and a controlled exercise program of 12 weeks duration. The horses were assessed 3, 6, 12, 24 weeks and one year after the first treatment. Of the 34 cases with forelimb PSD, 21 (61.8%) had returned to full work by six months after diagnosis and 19 cases (55.9%) were still in full work one year after ESWT. Of the 22 horses with hindlimb PSD, 9 (40.9%) had returned to full work by six months and 4 (18.2%) were still in full work one year after diagnosis. There was no association (chi-square test) between the outcome and the severity of the initial ultrasonographic and/or radiographic findings. Compared with the results of other clinical studies, these findings suggest that in horses with PSD of fore- and hindlimb, the prognosis for returning to full work six months after diagnosis can be improved when ESWT and a controlled exercise program are used. However, a high rate of recurrence occurred in the hindlimb despite treatment. PMID- 17076465 TI - Technetium-99m-HDP uptake characteristics in equine fractures: a retrospective study. AB - Bone scintigraphy is a very sensitive diagnostic tool to detect elevated bone metabolism. In cases of fractures and fissure fractures, the radiopharmaceutical uptake in the bone is said to be increased within a few hours after the injury. In this retrospective study, the scintigraphic uptake characteristics at the fracture site of 36 horses with radiographically confirmed fractures or fissure fractures were evaluated. Uptake ratios between the fracture region and adjacent normal bone or soft tissue activity respectively were calculated and compared to different anamnestic and radiographic data. The overall sensitivity of bone scintigraphy was 94.4% (34 positive cases out of 36). In the 36 horses, no correlation between the age of the fracture and the radiopharmaceutical uptake was found. However, there seems to be a lack of sensitivity in early detection of equine pelvic fractures when a standing bone scintigraphy examination protocol is used. PMID- 17076466 TI - Formation of FePt nanoparticles having high coercivity. AB - Ultrasonication of toluene solutions of the heteropolynuclear cluster complex, Pt3Fe3(CO)15, in the presence of oleic acid and oleylamine affords surface-capped fcc FePt nanoparticles having an average diameter of ca. 2 nm. Self-assembled arrays of these nanoparticles on oxidized Si wafers undergo a fcc-to-fct phase transition at 775 degrees C to form ferromagnetic FePt nanocrystals ca. 5.8 nm in diameter well dispersed on the Si wafer surface. Room-temperature coercivity measurements of these annealed FePt nanoparticles confirm a high coercivity of ca. 22.3 kOe. Such high coercivity for fct FePt nanoparticles might result from use of a heterpolynuclear complex as a single-source precursor of Fe and Pt neutral atoms or from use of ultrasonication to form fcc FePt nanoparticles under conditions of exceptionally rapid heating. Experiments to determine the critical experimental conditions required to achieve such high room-temperature coercivities in ferromagnetic nanoparticles are underway. PMID- 17076467 TI - Hydrogen bonding makes a difference in the rhodium-catalyzed enantioselective hydrogenation using monodentate phosphoramidites. AB - A new generation of monodentate phosphoramidite ligands bearing a primary amine moiety was found to display comparable or better efficiency than bisphosphines in the Rh-catalyzed asymmetric hydrogenation of challenging substrates, such as (Z) methyl alpha-acetoxyacrylate or (E)-beta-aryl itaconate derivatives, affording the corresponding hydrogenation products with excellent enantioselectivities (up to >99% ee). The presence of intermolecular hydrogen bonding (HB) between two monodentate ligands in the catalyst was found to be critical for excellent catalyst performance. This finding provides a basis for design and development of further catalyst systems using this type of monodentate phosphoramidite ligands. PMID- 17076468 TI - Modified pyrimidines specifically bind the purine riboswitch. AB - The purine riboswitch is a genetic regulatory element found in the 5' untranslated regions of Gram-positive bacteria that regulates expression of the mRNA specifically in response to either guanine or adenine. We report that the adenine-responsive RNA element is also capable of specifically recognizing pyrimidine compounds bearing modifications at the 6- or 5,6-positions in a fashion similar to that of purine compounds. Using isothermal titration calorimetry and X-ray crystallography, the binding of these compounds is characterized. PMID- 17076469 TI - Electron-transfer reduction of cup-stacked carbon nanotubes affording cup-shaped carbons with controlled diameter and size. AB - The electron-transfer reduction of cup-stacked carbon nanotubes (CSCNTs) with sodium naphthalenide and the subsequent treatment with 1-iodododecane results in electrostatically destacking of CSCNTs to afford cup-shaped carbons with controlled diameter and size. The dodecylated cup-shaped carbons were highly dispersed and stable in nonpolar solvents, such as THF, tetrachloroethylene, and chloroform, as compared with the pristine CSCNTs. PMID- 17076470 TI - A Heterotetranuclear [NiIIReIV3] single-molecule magnet. AB - The reaction of [ReIVCl4(ox)]2- and fully solvated Ni2+ ions in a MeCN/i-PrOH mixture affords the heterotetranuclear complex (NBu4)4[Ni{ReCl4(ox)}3] where the rhenium precursor acts as a bidentate ligand toward the nicke(II) ion through the oxalate group. The mixed 3d-5d species exhibits intramolecular ferromagnetic coupling and it behaves like a single-molecule magnet. PMID- 17076471 TI - sp3 C-H bond arylation directed by amidine protecting group: alpha-arylation of pyrrolidines and piperidines. AB - A new ruthenium-catalyzed direct sp3 C-H to C-C bond transformation is disclosed. Specifically, a method for the alpha-arylation of cyclic amines, containing either permanent (pyridine, pyrimidine) or removable (amidine) directing groups, is described. This cross-coupling reaction involves heating amine substrates with arylboronate ester at 150 degrees C in a ketone solvent with a catalytic amount of ruthenium carbonyl [Ru3(CO)12]. Arylboronate esters containing either electron withdrawing or electron-donating substituents could be efficiently coupled. Heteroarene boronates were also effective donors. PMID- 17076472 TI - A low-spin alkylperoxo-iron(III) complex with weak Fe-O and O-O bonds: implications for the mechanism of superoxide reductase. AB - The synthesis of a mononuclear, five-coordinate ferrous complex [([15]aneN4)FeII(SPh)](BF4) (1) is reported. This complex is a new model of the reduced active site of the enzyme superoxide reductase (SOR), which is comprised of a [(NHis)4(Scys)FeII] center. Complex 1 reacts with alkylhydroperoxides (tBuOOH, cumenylOOH) at low temperature to give a metastable, dark red intermediate (2a: R = tBu; 2b: R = cumenyl) that has been characterized by UV vis, EPR, and resonance Raman spectroscopy. The UV-vis spectrum (-80 degrees C) reveals a 526 nm absorbance (epsilon = 2150 M-1 cm-1) for 2a and a 527 nm absorbance (epsilon = 1650 M-1 cm-1) for 2b, indicative of alkylperoxo-to iron(III) LMCT transitions, and the EPR data (77 K) show that both intermediates are low-spin iron(III) complexes (g = 2.20 and 1.97). Definitive identification of the Fe(III)-OOR species comes from RR spectra, which give nu(Fe-O) = 612 (2a) and 615 (2b) cm-1, and nu(O-O) = 803 (2a) and 795 (2b) cm-1. The assignments for 2a were confirmed by 18O substitution (tBu18O18OH), resulting in a 28 cm-1 downshift for nu(Fe-18O), and a 46 cm-1 downshift for nu(18O-18O). These data show that 2a and 2b are low-spin FeIII-OOR species with weak Fe-O bonds and suggest that a low-spin intermediate may occur in SOR, as opposed to previous proposals invoking high-spin intermediates. PMID- 17076473 TI - Colchicine glycorandomization influences cytotoxicity and mechanism of action. AB - The reaction of 70 unprotected, diversely functionalized free reducing sugars with methoxyamine-appended colchicine led to the production of a 58-member glycorandomized library. High-throughput cytotoxicity assays revealed glycosylation to modulate specificity and potency. Library members were also identified which, unlike the parent natural product (a destabilizer), stabilized in vitro tubulin polymerization in a manner similar to taxol. This study highlights a simple extension of neoglycorandomization toward amine-bearing scaffolds and the potential benefit of glycosylating nonglycosylated natural products. PMID- 17076474 TI - Spectrally-resolved atomic-scale length variations of gold nanorods. AB - Functionality of gold nanorod structures as ultra-sensitive optical rulers is demonstrated. Arrays of gold nanorods were fabricated by electron beam lithography and lift-off techniques with high accuracy and uniformity. Their longitudinal plasmon scattering spectra were found to exhibit extreme sensitivity to the length of the nanorods. This phenomenon enables optical detection of the nanorod length variations comparable to the thickness of a few atomic layers of gold. PMID- 17076475 TI - La@C72 having a non-IPR carbon cage. AB - We show here that La@C72 has a non-IPR cage, unique electronic properties, and high reactivity by the spectroscopic and X-ray crystallographic analysis and the theoretical study. The isolation of La@C72 as a stable derivative might constitute an important stepping-stone on the way to isolation of these unknown metallofullerenes and open new material science of metallofullerenes. PMID- 17076476 TI - Identification of an "end-on" nickel-superoxo adduct, [Ni(tmc)(O2)]+. AB - An "end-on" Ni2+-superoxo adduct has been prepared via two independent synthetic routes and its structure ascertained by spectroscopic and computational methods. The new structure type in nickel coordination chemistry is supported by resonance Raman and EPR spectroscopic features, the former displaying a high frequency nu (O-O) mode (1131 cm-1) consistent with significant superoxo character. The Ni2+ superoxo adduct oxidizes PPh3 to OPPh3 in quantitative yield. PMID- 17076477 TI - Direct and high resolution characterization of cytochrome c equilibrium folding. AB - Protein folding has emerged as a central problem in biophysics, and the equilibrium folding mechanism of cytochrome c (cyt c) has served as a model system. Unfortunately, the detailed characterization of both the folding process and of any intermediate that might be populated has been limited by the low structural and/or temporal resolution of the available techniques. Here, we report the use of a recently developed technique to study folding that is based on the site-selective incorporation of carbon-deuterium (C-D) bonds and their characterization by IR spectroscopy. Specifically, we synthesize and characterize the protein with deuterated residues spread throughout four structural motifs: (d3)Leu68 in the 60's helix, (d8)Lys72 and (d8)Lys73 in the 70's helix, (d8)Lys79, (d3)Met80, and (d3)Ala83 in the D-loop, and (d3)Leu94, (d3)Leu98, and (d3)Ala101 in the C-terminal helix. The data reveal correlated behavior of the residues within each structural motif, as well as between the residues of the 60's and C-terminal helices and between residues of the 70's helix and D-loop. Residues of the 70's helix and the D-loop are more stable than those within the 60's and C-terminal helices, although the former are more sensitive to added denaturant. The data also suggest that the hydrophobicity of the heme cofactor plays a central role in folding. These results contrast with those from previous H/D exchange studies and suggest that the low denaturant fluctuations observed in the H/D exchange experiments are not similar to those through which the protein actually unfolds. The inherently fast time scale of IR also allows us to characterize the folding intermediate, long thought to be present, but which has proven difficult to characterize by other techniques. PMID- 17076478 TI - Mesoscale chiroptics of rhythmic precipitates. AB - Rhythmic precipitates of centrosymmetric phthalic acid were analyzed by a square wave mechanically modulated circular extinction imaging microscope. Spherulites were bisected into square-millimeter sized heterochiral domains that are a consequence of circular intensity differential scattering of left and right circularly polarized light. The dissymmetry and chiral amplification indicated optically was confirmed in the microtexture established by atomic force and scanning electron microscopies. PMID- 17076479 TI - Interplay of fidelity, binding strength, and structure in supramolecular polymers. AB - In studies of a supramolecular network of polymers formed by self-association of UPy or UG recognition units displayed along a poly(butyl methacrylate) (PBMA) backbone, it was unexpectedly found that the more weakly dimerizing (Kdimer approximately 200 M-1) UG unit produced more assembly than did the very strongly dimerizing UPy unit (Kdimer = 2 x 107 M-1). Likewise, in examining supramolecular blends mediated by the heterocomplexation of DAN and UPy, which occurs upon the mixing of polystyrene containing the DAN unit (PS-DAN) and PBMA-UPy, increasing the mol % of UPy did not produce increased viscosity. 1H NMR showed that both observations can be explained by the intramolecular recognition of UPy. Structural studies show that the length of the chain linking the UPy unit to the backbone is critical, with longer linkers favoring intermolecular dimers. An interplay of linker chain length, polymer Mw, recognition unit mol %, and fidelity determines the extent of network growth. PMID- 17076480 TI - Design of open-framework germanates by combining different building units. AB - Open-framework germanates with extra-large channels are built from two different types of clusters. The two different clusters are connected in such a way to form a pseudo-body-centered cluster aggregate (PBCCA). The different packing of PBCCAs resulted in different 3D open-framework germanates. PMID- 17076481 TI - Channel-selective independent sorption and collection of hydrophilic and hydrophobic molecules by Cs2[Cr3O(OOCC2H5)6(H2O)3]2[alpha-SiW12O40] ionic crystal. AB - An ionic crystal of Cs2[Cr3O(OOCC2H5)6(H2O)3]2[alpha-SiW12O40].4H2O 1a with hydrophilic and hydrophobic channels has been designed and successfully synthesized. The guest-free phase 1b sorbs dichloromethane and water in the hydrophobic and hydrophilic channels, respectively. The rate and equilibrium amount of the dichloromethane sorption into the hydrophobic channel and those of water into the hydrophilic channel were independent of each other. The sorption properties can be applied to the channel-selective sorption and collection of hydrophobic (dichloromethane) and hydrophilic (water) molecules from the mixture. PMID- 17076482 TI - Control of absolute helicity in single-stranded abiotic metallofoldamers. AB - Described is the design, synthesis, and characterization of abiotic, single stranded metallofoldamers that adopt helical secondary structures upon metal complexation. The absolute helicity is determined by stereocenters at the ends of the structures and is enforced by the steric influence and hydrogen bonding ability of esters in the backbone of the foldamer. Folding of the structures is characterized in the solid state by X-ray crystallography and in solution by specific rotation, CD spectroscopy, and NMR spectroscopy. PMID- 17076483 TI - Effect of nanoscale geometry on molecular conformation: vibrational sum-frequency generation of alkanethiols on gold nanoparticles. AB - Vibrational sum frequency generation (VSFG) spectroscopy was used to study the nanoscale geometric effects on molecular conformation of dodecanethiol ligand on gold nanoparticles of varying size between 1.8 and 23 nm. By analyzing the CH3 and CH2 stretch transitions of dodecanethiol using the spectroscopic propensity rules for the SFG process, we observe the increase of the gauche defects in the alkyl chain of the ligand on the nanoparticle surface when the curvature approaches the size of the molecule ( approximately 1.6 nm). In contrast, linear infrared absorption and Raman spectra, governed by different selection rules, do not allow observation of the size-dependent conformational changes. The results are understood in terms of the geometric packing effect, where the curvature of the nanoparticle surface results in the increased conical volume available for the alkyl chain. PMID- 17076484 TI - Heme-heme interactions in the cytochrome b6f complex: EPR spectroscopy and correlation with structure. AB - Cytochrome b6f of oxygenic photosynthesis was studied using multifrequency, multimode EPR Spectroscopy. Frequency dependent signals above g = 4.3, and the observation of parallel-mode signals, are indicative of spin interactions in the complex. We demonstrate the presence of an exchange interaction between the unique high-spin heme cn and a nearby low-spin heme bn, and show that a quinone analog NQNO binds at or near to heme cn. The two hemes remain spin coupled upon the binding of NQNO, though strength of interaction decreases significantly. The electronic coupling implies that the heme bn/cn pair could function as a unit to facilitate 2-electron reduction of plastoquionone without generation of an energetically unfavorable semiquinone intermediate. PMID- 17076485 TI - Fluoride ion capture from water with a cationic borane. AB - The reaction of the [Li(THF)4]+ salt of dimesityl-1,8-naphthalenediylborate with [Me2NCH2]I affords a borane (1-(Mes2B)-8-(Me2NCH2)-C10H6) which can be converted into a cationic borane [3]+ ([1-(Mes2B)-8-(Me3NCH2)-C10H6]+) by methylation with MeOTf. This cationic borane promptly complexes fluoride to afford the corresponding zwitterionic ammonium/fluoroborate 3-F (1-(Mes2FB)-8-(Me3NCH2) C10H6). Cation [3]+ fails to react with chloride, bromide, and iodide indicating that fluoride complexation is selective. Structural, spectroscopic, and computational studies carried out on 3-F show the existence of an unusual C-H...F B hydrogen bond. Remarkably, [3]+ captures fluoride from water under biphasic conditions (H2O/CHCl3) to form 3-F. The high fluoride affinity of [3]+ can be correlated to the Coulombic forces which stabilize the B-F bond against heterolysis. PMID- 17076486 TI - Photodeformable spherical hybrid nanoparticles. AB - We report the first synthesis of spherical nanoparticles of a bridged polysilsesquioxane. The hard, brittle nanoparticles are prepared by a simple sol gel polymerization without surfactants or templates. Particle size ranges from 50 to 100 nm. The organic component of these hybrid nanoparticles is composed of a photolabile coumarin dimer. Irradiation with UV light dissociates the photodimer resulting in rupture of the cross-links and subsequent deformation and eventual "melting" of the nanoparticles. PMID- 17076487 TI - Antimicrobial leucocin analogues with a disulfide bridge replaced by a carbocycle or by noncovalent interactions of allyl glycine residues. AB - The type IIa bacteriocins are antimicrobial peptides isolated from lactic acid bacteria that act as food preservation agents and have nanomolar activity against pathogens such as Listeria monocytogenes. Previous reports with mutant bacteriocins indicate that the conserved disulfide bridge between cysteine residues 9 and 14 in bacteriocins such as leucocin A (1) is critical for antibiotic properties, which are mediated by target membrane receptor proteins belonging to the mannose phosphotransferase (mpt) system. To examine whether the disulfide can be replaced by an olefin moiety, [9,14]-dicarba leucocin A (4) was made by on-resin ring closing metathesis of allyl glycine residues using a new protocol suitable for larger hydrophobic peptides. Carbocyclic analogue 4 still displays nanomolar activity but is about 10-fold less potent than 1. Surprisingly, the acyclic [9,14]-diallyl leucocin A (5) displays even higher antibiotic activity than 4 and is as effective as the parent, leucocin A (1). We attribute this activity to hydrophobic intermolecular interactions of the diallyl side chains of the acyclic bacteriocin 5 that assist realization of the correct conformation at the receptor active site. Such substitutions in other systems may allow linear acyclic peptides to mimic the biological activity of natural disulfide ring-containing parents. PMID- 17076488 TI - Single-step synthesis of pyrimidine derivatives. AB - We describe a single-step conversion of various N-vinyl and N-aryl amides to the corresponding pyrimidine and quinazoline derivatives, respectively. The process involves amide activation with 2-chloropyridine and trifluoromethanesulfonic anhydride followed by nitrile addition into the reactive intermediate and cycloisomerization. In situ nitrile generation from primary amides allows for their use as nitrile surrogates. The use of this chemistry with sensitive N-vinyl amides and epimerizable substrates in addition to a wide range of functional groups is noteworthy. PMID- 17076489 TI - Alkyl-functionalized organic dyes for efficient molecular photovoltaics. AB - We designed and synthesized new alkyl-functionalized organic dyes, MK-1 and MK-2, for dye-sensitized solar cells (DSSCs). Based on the MK-2 dye, a high performance of efficiency (eta, 7.7%; short-circuit current density Jsc = 14.0 mA cm-2, open circuit voltage Voc = 0.74 V, and fill factor FF = 0.74) was achieved under AM 1.5 G irradiation (100 mW cm-2). Remarkably, the relatively higher Voc for DSSCs based on MK-1 and MK-2 dyes, which have long alkyl chains, were observed among the organic dyes caused by the increasing of the electron lifetime in the conduction band of TiO2. Our molecular design of alkyl-functionalized dyes strongly suggests the promising performance of molecular photovoltaics based on organic dyes. PMID- 17076490 TI - Freestanding mesoporous quasi-single-crystalline CO3O4 nanowire arrays. AB - We report a facile template-free method for the large-area growth of freestanding hollow Co3O4 nanowire arrays on a variety of substrates including transparent conducting glass, Si wafer, and copper foil, et al. These nanowires have the interesting combined properties of mesoporosity and quasi-single-crystallinity. With their high surface area and crystallinity, and their direct growth on conductive substrate, these Co3O4 nanowire arrays will have promising applications in lithium-ion batteries, chemical sensing, and field-emission and electrochromic devices. Using the prepared nanowire arrays as electrode, an electrochemical sensor for hydrogen peroxide sensing has been demonstrated. PMID- 17076491 TI - Control of pyramidal inversion rates by redox switching. AB - The dynamics of pyramidal nitrogen inversion can be controlled by reversible redox switching in trans-2,3-diphenylaziridines bearing a suitable 1,4 naphthaquinone substituent. In the reduced form, an intramolecular H-bond significantly raises the inversion barrier slowing this molecular motion by >50 fold. The experimental findings are further supported by DFT calculations. PMID- 17076492 TI - Characterization of SpnQ from the spinosyn biosynthetic pathway of Saccharopolyspora spinosa: mechanistic and evolutionary implications for C-3 deoxygenation in deoxysugar biosynthesis. AB - The C-3 deoxygenation step in the biosynthesis of d-forosamine (4-N,N dimethylamino-2,3,4,6-tetradeoxy-d-threo-hexopyranose), a constituent of spinosyn produced by Saccharopolyspora spinosa, was investigated. The spnQ gene, proposed to encode a TDP-4-keto-2,6-dideoxy-d-glucose 3-dehydratase was cloned and overexpressed in E. coli. Characterization of the purified enzyme established that it is a PMP and iron-sulfur containing enzyme which catalyzes the C-3 deoxygenation in a reductase-dependent manner similar to that of the previously well characterized hexose 3-dehydrase E1 from Yersinia pseudotuberculosis. However, unlike E1, which has evolved to work with a specific reductase partner present in its gene cluster, SpnQ lacks a specific reductase, and works efficiently with general cellular reductases ferredoxin/ferredoxin reductase or flavodoxin/flavodoxin reductase. SpnQ also catalyzes C-4 transamination in the absence of an electron transfer intermediary and in the presence of PLP and l glutamate. Under the same conditions, both E1 and the related hexose 3-dehydrase, ColD, catalyze C-3 deoxygenation. Thus, SpnQ possesses important features which distinguish it from other well studied homologues, suggesting unique evolutionary pathways for each of the three hexose 3-dehydrases studied thus far. PMID- 17076494 TI - Surface-mediated NH and N addition to styrene on Ag(110). AB - In this paper, we report the investigation of the reaction of adsorbed NH and N with styrene on Ag(110) using temperature-programmed reaction spectroscopy. Using O2 and NH3 as the starting reagents, NH and N species were deposited on a Ag surface. The reaction of styrene on NH- and N-covered Ag surface appears to yield 2-phenylaziridine and benzonitrile, with additional products HCN and NH3. The formation of aziridine, the nitrogenous analogue of styrene epoxide, is proposed to be due to the cycloaddition of adsorbed NH to the carbon-carbon double bond. These results suggest that Ag-based heterogeneous catalysts may be useful for the aziridination of olefins. PMID- 17076493 TI - Kinetic resolutions of indolines by a nonenzymatic acylation catalyst. AB - The first method for the kinetic resolution of indolines through catalytic N acylation is described. To improve the selectivity factor, new planar-chiral PPY derived catalysts were synthesized, wherein the chiral environment was systematically modified. This work provides a rare example of a nonenzyme-based acylation catalyst for the kinetic resolution of amines. PMID- 17076495 TI - Stereoselective preparation of 1-siloxy-1-alkenylcopper species by 1,2-Csp2-to-O silyl migration of acylsilanes. AB - 1-Siloxy-1-alkenylcopper species were generated by 1,2-Csp2-to-O silyl migration of the copper enolates of acyltriphenylsilanes. The alkenylcopper species reacted with methyl, benzyl, allylic, and tributylstannyl halides to give geometrically pure (Z)-enol silyl ethers. In the presence of Pd(0) catalyst, the cross-coupling of the alkenyl copper species with aryl and alkenyl iodides also proceeded to give the (Z)-enol silyl ethers with high stereoselectivity. PMID- 17076496 TI - Organolutetium vinyl and tuck-over complexes via C-H bond activation. AB - The vinyl C-H bond of tetramethylfulvene is activated in the presence of [(C5Me5)2LuH]x, 1, to form a vinyl organolutetium complex, (C5Me5)2Lu(CH=C5Me4), 2. Also formed in the reaction is the "tuck-over" complex, (C5Me5)2Lu(mu-H)(mu eta1:eta5-CH2C5Me4)Lu(C5Me5), 3, containing a (CH2C5Me4)2- moiety long postulated to exist in organolutetium chemistry but never crystallographically characterized. Evidence for these C-H bond activations by a "(C5Me5)3Lu" intermediate, 4, is presented. Complex 3 can also be made in high yield by thermolysis of 1. Under H2, 1 catalytically hydrogenates TMF to C5Me5H. PMID- 17076497 TI - Dramatic solvent and hydration effects on the transition state of soybean peroxidase. AB - Enzymes are shown to function in nonaqueous media; however, relatively little information is available on the influence of the organic solvent as well as its associated water content on the properties of the enzymatic transition states. A better understanding of these effects will be useful in developing kinetic models that can then be used to predict optimal solvent and substrate choices for enzymatic reactions in organic media. The influence of the reaction media on soybean peroxidase-catalyzed oxidation of para-substituted phenols was studied using Hammett analysis for several organic solvent systems. The catalytic activity and substrate specificity of the enzyme are influenced by the nature of the solvent and its associated hydration. These findings may allow one to draw conclusions about the reaction mechanism and the roles of solvent and solvent hydration on enzyme function. PMID- 17076498 TI - A gold-catalyzed unique cycloisomerization of 1,5-enynes: efficient formation of 1-carboxycyclohexa-1,4-dienes and carboxyarenes. AB - A novel Au-catalyzed migratory cycloisomerization strategy is advanced. Implementation of this strategy led to the development of a unique Au-catalyzed 1,5-enyne cycloisomerizatioin involving carboxy migration and Au-mediated C-C single bond formation. 1-Carboxycyclohexa-1,4-dienes and carboxyarenes can be prepared with good efficiency and with flexible substitution patterns. PMID- 17076499 TI - Directed biosynthesis of alkaloid analogs in the medicinal plant Catharanthus roseus. AB - Terpene indole alkaloids are plant natural products with diverse structures and biological activities. A highly branched biosynthetic pathway is responsible for the production of approximately 130 different alkaloids in Madagascar periwinkle (C. roseus) from a common biosynthetic intermediate derived from tryptamine. Although numerous biosynthetic pathways can incorporate unnatural starting materials to yield novel natural products, it was not clear how efficiently the complex, eukaryotic TIA pathway could utilize unnatural substrates to make new alkaloids. This work demonstrates that the TIA biosynthetic machinery can be used to produce novel alkaloid structures and also highlights the potential of this pathway for future metabolic engineering efforts. PMID- 17076500 TI - A general synthetic strategy for oxide-supported metal nanoparticle catalysts. AB - Despite recent exciting progress in catalysis by supported gold nanoparticles, there remains the formidable challenge of preparing supported gold catalysts that collectively incorporate precise control over factors such as size and size distribution of the gold nanoparticles, homogeneous dispersion of the particles on the support, and the ability to utilize a wide range of supports that profoundly affect catalytic performance. Here, we describe a synthetic methodology that achieves these goals. In this strategy, weak interface interactions evenly deposit presynthesized organic-capped metal nanoparticles on oxide supports. The homogeneous dispersion of nanoparticles on oxides is then locked in place, without aggregation, through careful calcination. The approach takes advantage of recent advances in the synthesis of metal and oxide nanomaterials and helps to bring together these two classes of materials for catalysis applications. An important feature is that the strategy allows metal nanoparticles to be well dispersed on a variety of oxides with few restrictions on their physical and chemical properties. Following this synthetic procedure, we have successfully developed efficient gold catalysts for green chemistry processes, such as the production of ethyl acetate from the selective oxidation of ethanol by oxygen at 100 degrees C. PMID- 17076501 TI - Mechanistic borderline between one-step hydrogen transfer and sequential transfers of electron and proton in reactions of NADH analogues with triplet excited states of tetrazines and Ru(bpy)(3)2+. AB - Efficient energy transfer from Ru(bpy)(3)(2+) (bpy = 2,2'-bipyridine, denotes the excited state) to 3,6-disubstituted tetrazines [R(2)Tz: R = Ph (Ph(2)Tz), 2 chlorophenyl [(ClPh)(2)Tz], 2-pyridyl (Py(2)Tz)] occurs to yield the triplet excited states of tetrazines ((3)R(2)Tz(*)), which have longer lifetimes and higher oxidizing ability as compared with those of Ru(bpy)(3)(2+). The dynamics of hydrogen-transfer reactions from NADH (dihydronicotinamide adenine dinucleotide) analogues has been examined in detail using (3)R(2)Tz(*) by laser flash photolysis measurements. Whether formal hydrogen transfer from NADH analogues to (3)R(2)Tz(*) proceeds via a one-step process or sequential electron and proton transfer processes is changed by a subtle difference in the electron donor ability and the deprotonation reactivity of the radical cations of NADH analogues as well as the electron-acceptor ability of (3)R(2)Tz(*) and the protonation reactivity of R(2)Tz(*)(-). In the case of (3)Ph(2)Tz(*), which is a weaker electron acceptor than the other tetrazine derivatives [(ClPh)(2)Tz; Py(2)Tz], direct one-step hydrogen transfer occurs from 10-methyl-9,10 dihydroacridine (AcrH(2)) to (3)Ph(2)Tz(*) without formation of the radical cation (AcrH(2)(*)(+)). The rate constant of the direct hydrogen transfer from AcrH(2) to (3)Ph(2)Tz(*) is larger than that expected from the Gibbs energy relation for the rate constants of electron transfer from various electron donors to (3)Ph(2)Tz(*), exhibiting the primary deuterium kinetic isotope effect. On the other hand, hydrogen transfer from 9-isopropyl-10-methyl-9,10-dihydroacridine (AcrHPr(i)) and 1-benzyl-1,4-dihydronicotinamide (BNAH) to (3)R(2)Tz(*) occurs via sequential electron and proton transfer processes, when both the radical cations and deprotonated radicals of NADH analogues are detected by the laser flash photolysis measurements. PMID- 17076502 TI - Mechanistic study of hydrogen transfer to imines from a hydroxycyclopentadienyl ruthenium hydride. Experimental support for a mechanism involving coordination of imine to ruthenium prior to hydrogen transfer. AB - Reaction of [2,3,4,5-Ph(4)(eta(5)-C(4)COH)Ru(CO)(2)H] (2) with different imines afforded ruthenium amine complexes at low temperatures. At higher temperatures in the presence of 2, the complexes decomposed to give [Ru(2)(CO)(4)(mu H)(C(4)Ph(4)COHOCC(4)Ph(4))] (1) and free amine. Electron-rich imines gave ruthenium amine complexes with 2 at a lower temperature than did electron deficient imines. The negligible deuterium isotope effect (k(RuHOH)/k(RuDOD) = 1.05) observed in the reaction of 2 with N-phenyl[1-(4 methoxyphenyl)ethylidene]amine (12) shows that neither hydride (RuH) nor proton (OH) is transferred to the imine in the rate-determining step. In the dehydrogenation of N-phenyl-1-phenylethylamine (4) to the corresponding imine 8 by [2,3,4,5-Ph(4)(eta(4)-C(4)CO)Ru(CO)(2)] (A), the kinetic isotope effects observed support a stepwise hydrogen transfer where the isotope effect for C-H cleavage (k(CHNH)/k(CDNH) = 3.24) is equal to the combined (C-H, N-H) isotope effect (k(CHNH)/k(CDND) = 3.26). Hydrogenation of N-methyl(1 phenylethylidene)amine (14) by 2 in the presence of the external amine trap N methyl-1-(4-methoxyphenyl)ethylamine (16) afforded 90-100% of complex [2,3,4,5 Ph(4)(eta(4)-C(4)CO)]Ru(CO)(2)NH(CH(3))(CHPhCH(3)) (15), which is the complex between ruthenium and the amine newly generated from the imine. At -80 degrees C the reaction of hydride 2 with 4-BnNH-C(6)H(9)=NPh (18), with an internal amine trap, only afforded [2,3,4,5-Ph(4)(eta(4)-C(4)CO)](CO)(2)RuNH(Ph)(C(6)H(10)-4 NHBn) (19), where the ruthenium binds to the amine originating from the imine, showing that neither complex A nor the diamine is formed. Above -8 degrees C complex 19 rearranged to the thermodynamically more stable [Ph(4)(eta(4) C(4)CO)](CO)(2)RuNH(Bn)(C(6)H(10)-4-NHPh) (20). These results are consistent with an inner sphere mechanism in which the substrate coordinates to ruthenium prior to hydrogen transfer and are difficult to explain with the outer sphere pathway previously proposed. PMID- 17076503 TI - DFT study on the palladium-catalyzed allylation of primary amines by allylic alcohol. AB - The palladium-catalyzed allylation of primary amines has been investigated by DFT calculations (B3PW91, PCM method), and two potential mechanisms were studied. The first mechanism relies on the formation of cationic hydridopalladium complexes. Their formation involves a metal-assisted formal (1,3) shift of a proton from the nitrogen atom of an ammonium to the Cbeta carbon atom. The second part of the cycle relies on a ligand exchange through a pentacoordinated 18VE hydridopalladium complex. The last step likely proceeds through a bimolecular pathway and formally consists of a proton transfer from the allylammonium to the alcohol group of the complex. The second mechanism, which is closer to that currently admitted for nucleophilic allylic substitutions, relies on the decomplexation of the coordinated allylammonium and appears to be favored. This catalytic cycle was recomputed on model complexes varying the ligands, and a charge decomposition analysis was carried out to assess the influence of the electronic properties of the ligands. To compare our results with competitive experiments, CDA calculations were also performed on real ligands. In agreement with experimental observations, this process was found to be strongly ligand dependent, decomplexation being favored by strong pi-acceptor ligands. These calculations led us to show experimentally that complex [Pd(P(OPh)(3))(2)(eta(3) C(3)H(5))][OTf] is an efficient catalyst for this allylation. Finally, this catalytic process proved to be sensitive to the nature of the amine, with poorly basic amines favoring the re-formation of the catalytic precursor. PMID- 17076504 TI - Picosecond isomerization in photochromic ruthenium-dimethyl sulfoxide complexes. AB - The complexes [Ru(tpy)(bpy)(dmso)](OSO(2)CF(3))(2) and trans [Ru(tpy)(pic)(dmso)](PF(6)) (tpy is 2,2':6',2' '-terpyridine, bpy is 2,2' bipyridine, pic is 2-pyridinecarboxylate, and dmso is dimethyl sulfoxide) were investigated by picosecond transient absorption spectroscopy in order to monitor excited-state intramolecular S-->O isomerization of the bound dmso ligand. For [Ru(tpy)(bpy)(dmso)](2+), global analysis of the spectra reveals changes that are fit by a biexponential decay with time constants of 2.4 +/- 0.2 and 36 +/- 0.2 ps. The first time constant is assigned to relaxation of the S-bonded (3)MLCT excited state. The second time constant represents both excited-state relaxation to ground state and excited-state isomerization to form O [Ru(tpy)(bpy)(dmso)](2+). In conjunction with the S-->O isomerization quantum yield (Phi(S)(-->)(O) = 0.024), isomerization of [Ru(tpy)(bpy)(dmso)](2+) occurs with a time constant of 1.5 ns. For trans-[Ru(tpy)(pic)(dmso)](+), global analysis of the transient spectra reveals time constants of 3.6 +/- 0.2 and 118 +/- 2 ps associated with these two processes. In conjunction with the S-->O isomerization quantum yield (Phi(S)(-->)(O) = 0.25), isomerization of trans [Ru(tpy)(pic)(dmso)](+) occurs with a time constant of 480 ps. In both cases, the thermally relaxed excited states are assigned as terpyridine-localized (3)MLCT states. Electronic state diagrams are compiled employing these data as well as electrochemical, absorption, and emission data to describe the reactivity of these complexes. The data illustrate that rapid bond-breaking and bond-making reactions can occur from (3)MLCT excited states formed from visible light irradiation. PMID- 17076505 TI - On the biosynthetic origin of methoxymalonyl-acyl carrier protein, the substrate for incorporation of "glycolate" units into ansamitocin and soraphen A. AB - Feeding experiments with isotope-labeled precursors rule out hydroxypyruvate and TCA cycle intermediates as the metabolic source of methoxymalonyl-ACP, the substrate for incorporation of "glycolate" units into ansamitocin P-3, soraphen A, and other antibiotics. They point to 1,3-bisphosphoglycerate as the source of the methoxymalonyl moiety and show that its C-1 gives rise to the thioester carbonyl group (and hence C-1 of the "glycolate" unit), and its C-3 becomes the free carboxyl group of methoxymalonyl-ACP, which is lost in the subsequent Claisen condensation on the type I modular polyketide synthases (PKS). d-[1,2 (13)C(2)]Glycerate is also incorporated specifically into the "glycolate" units of soraphen A, but not of ansamitocin P-3, suggesting differences in the ability of the producing organisms to activate glycerate. A biosynthetic pathway from 1,3 bisphosphoglycerate to methoxymalonyl-ACP is proposed. Two new syntheses of R- and S-[1,2-(13)C(2)]glycerol were developed as part of this work. PMID- 17076506 TI - Stabilization of a kinetically favored nanostructure: surface ROMP of self assembled conductive nanocoils from a norbornene-appended hexa-peri hexabenzocoronene. AB - Newly designed norbornene-appended hexabenzocoronene 1 self-assembles, upon diffusion of an Et(2)O vapor into its CH(2)Cl(2) solution, to form either graphitic nanocoils or nanotubes, depending on the self-assembling conditions. The coiled assembly, selectively formed at 15 degrees C, is a kinetic intermediate for the tubular assembly and transforms into nanotubes on standing at 25 degrees C. However, post-ring-opening metathesis polymerization of the norbornene pendants of 1 enhances the thermal stability of the coiled assembly as well as the tubular one and disables a thermodynamic coil-to-tube transition. The polymerized nanocoils show an electroconductivity of 1 x 10(-)(4) S cm(-)(1) upon doping with I(2), while the nonpolymerized nanocoils are disrupted upon being doped. PMID- 17076507 TI - Ion permeability of SAMs on nanoparticle surfaces. AB - This article reports on the ion permeability of self-assembled monolayers (SAMs) formed on the surface of charged alkanethiol-protected gold nanoparticles, so called monolayer-protected clusters (MPCs). The capacitance and thus the charging energy required to add/remove an electron from the metal core are extremely sensitive to ions entering the monolayer, and the extent of ion penetration can be tuned by the charge and size of the ions and the permittivity of the solvent. Experimentally, this effect is comparable to ion association with conventional redox molecules, indicating that MPCs despite their large size and the fundamentally differing nature of the electron transfer process can be treated analogously to redox molecules. PMID- 17076508 TI - Nanostructures versus solid solutions: low lattice thermal conductivity and enhanced thermoelectric figure of merit in Pb9.6Sb0.2Te10-xSex bulk materials. AB - The series of Pb(9.6)Sb(0.2)Te(10)(-)(x)Se(x) compounds with different Se content (x) were prepared, and their structure was investigated at the atomic and nanosized regime level. Thermoelectric properties were measured in the temperature range from 300 to 700 K. The Pb(9.6)Sb(0.2)Te(10)(-)(x)Se(x) series was designed after the refinement of the single-crystal structure of Pb(3.82)Sb(0.12)Te(4) (Pb(9.6)Sb(0.3)Te(10); S.G. Pmm) by substituting isoelectronically in anion positions Te by Se. The Pb(9.6)Sb(0.2)Te(10)( )(x)Se(x)() compounds show significantly lower lattice thermal conductivity (kappa(L)) compared to the well-known PbTe(1)(-)(x)Se(x) solid solutions. For Pb(9.6)Sb(0.2)Te(3)Se(7) (x = 7), a kappa(L) value as low as 0.40 W/m.K was determined at 700 K. High-resolution transmission electron microscopy of several Pb(9.6)Sb(0.2)Te(10)(-)(x)Se(x) samples showed widely distributed Sb-rich nanocrystals in the samples which is the key feature for the strong reduction of the lattice thermal conductivity. The reduction of kappa(L) results in a significantly enhanced thermoelectric figure of merit of Pb(9.6)Sb(0.2)Te(10)( )(x)Se(x) compared to the corresponding PbTe(1)(-)(x)Se(x) solid solution alloys. For Pb(9.6)Sb(0.2)Te(3)Se(7) (x = 7), a maximum figure of merit of ZT approximately 1.2 was obtained at approximately 650 K. This value is about 50% higher than that of the state-of-the-art n-type PbTe. The work provides experimental validation of the theoretical concept that embedded nanocrystals can promote strong scattering of acoustic phonons. PMID- 17076509 TI - A minimalist approach toward protein recognition by epitope transfer from functionally evolved beta-sheet surfaces. AB - New approaches for identifying small molecules that specifically target protein surfaces as opposed to active site clefts are of much current interest. Toward this goal, we describe a three-step methodology: in step one, we target a protein of interest by directed evolution of a small beta-sheet scaffold; in step two, we identify residues on the scaffold that are implicated in binding; and in step three, we transfer the chemical information from the beta-sheet to a small molecule mimic. As a case study, we targeted the proteolytic enzyme thrombin, involved in blood coagulation, utilizing a library of beta-sheet epitopes displayed on phage that were previously selected for conservation of structure. We found that the thrombin-binding, beta-sheet displaying mini-proteins retained their structure and stability while inhibiting thrombin at low micromolar inhibition constants. A conserved dityrosine recognition motif separated by 9.2 A was found to be common among the mini-protein inhibitors and was further verified by alanine scanning. A molecule containing two tyrosine residues separated by a linker that matched the spacing on the beta-sheet scaffold inhibited thrombin, whereas a similar dityrosine molecule separated by a shorter 6 A linker could not. Moreover, kinetic analysis revealed that both the mini-protein as well as its minimalist mimic with only two functional residues exhibited noncompetitive inhibition of thrombin. Thus, this reductionist approach affords a simple methodology for transferring information from structured protein scaffolds to yield small molecule leads for targeting protein surfaces with novel mechanisms of action. PMID- 17076510 TI - Direct nitric oxide detection in aqueous solution by copper(II) fluorescein complexes. AB - A series of FL(n) (n = 1-5) ligands, where FL(n) is a fluorescein modified with a functionalized 8-aminoquinoline group as a copper-binding moiety, were synthesized, and the chemical and photophysical properties of the free ligands and their copper complexes were investigated. UV-visible spectroscopy revealed a 1:1 binding stoichiometry for the Cu(II) complexes of FL(1), FL(3), and FL(5) in pH 7.0 buffered aqueous solutions. The reactions of FL(2) or FL(4) with CuCl(2), however, appear to produce a mixture of 1:1 and 1:2 complexes, as suggested by Job's plots. These binding modes were modeled by the synthesis and X-ray crystal structure determination of Cu(II) complexes of 2-[(quinolin-8 ylamino)methyl]phenol (modL), employed as a surrogate of the FL(n) ligand family. Two kinds of crystals, [Cu(modL)(2)](BF(4))(2) and [Cu(2)(modL')(2)(CH(3)OH)](BF(4))(2) (modL' = 2-[(quinolin-8 ylamino)methyl]phenolate), were obtained. The structures suggest that one oxygen and two nitrogen atoms of the FL(n) ligands most likely bind to Cu(II). Introduction of nitric oxide (NO) to pH 7.0 buffered aqueous solutions of Cu(FL(n)) (1 microM CuCl(2) and 1 microM FL(n)) at 37 degrees C induces an increase in fluorescence. The fluorescence response of Cu(FL(n)) to NO is direct and specific, which is a significant improvement over commercially available small molecule-based probes that are capable of detecting NO only indirectly. The NO-triggered fluorescence increase of Cu(FL(5)) occurs by reduction of Cu(II) to Cu(I) with concomitant dissociation of the N-nitrosated fluorophore ligand from copper. Spectroscopic and product analyses of the reaction of the FL(5) copper complex with NO indicated that the N-nitrosated fluorescein ligand (FL(5)-NO) is the species responsible for fluorescence turn-on. Density functional theory (DFT) calculations of FL(5) versus FL(5)-NO reveal how N-nitrosation of the fluorophore ligand brings about the fluorescence increase. The copper-based probes described in the present work form the basis for real-time detection of nitric oxide production in living cells. PMID- 17076511 TI - Synthesis of nonspherical colloidal particles with anisotropic properties. AB - We describe a promising and flexible technique for fabricating uniform nonspherical particles with anisotropic phase and surface properties. Our approach is based on the seeded polymerization technique in which monomer-swollen particles are polymerized. The polymerization causes a phase separation to occur, giving rise to two-phase nonspherical particles. We show that the elastic contraction of the swollen polymer particles induced by elevated polymerization temperatures plays an important role in the phase separation. Moreover, chemical anisotropy of nonspherical particles can be obtained by using immiscible polymer pairs and by employing surface treatments. Furthermore, we are able to produce amphiphilic dumbbell particles consisting of two different bulbs: hydrophilic poly (ethylene imine)-coated polystyrene and hydrophobic polystyrene. Controlled geometries of these amphiphilic nonspherical particles will allow a wide range of potential applications, such as engineered colloid surfactants. PMID- 17076512 TI - Tuning peptoid secondary structure with pentafluoroaromatic functionality: a new design paradigm for the construction of discretely folded peptoid structures. AB - Peptoids, or oligomers of N-substituted glycine, are an important class of non native polymers whose close structural similarity to natural alpha-peptides and ease of synthesis offer significant advantages for the study of biomolecular interactions and the development of biomimetics. Peptoids that are N-substituted with alpha-chiral aromatic side chains have been shown to adopt either helical or "threaded loop" conformations, depending upon solvent and oligomer length. Elucidation of the factors that impact peptoid conformation is essential for the development of general rules for the design of peptoids with discrete and novel structures. Here, we report the first study of the effects of pentafluoroaromatic functionality on the conformational profiles of peptoids. This work was enabled by the synthesis of a new, alpha-chiral amine building block, (S)-1 (pentafluorophenyl)ethylamine (S-2), which was found to be highly compatible with peptoid synthesis (delivering (S)-N-(1-(pentafluorophenyl)ethyl)glycine oligomers). The incorporation of this fluorinated monomer unit allowed us to probe both the potential for pi-stacking interactions along the faces of peptoid helices and the role of side chain electrostatics in peptoid folding. A series of homo- and heteropeptoids derived from S-2 and non-fluorinated, alpha-chiral aromatic amide side chains were synthesized and characterized by circular dichroism (CD) and nuclear magnetic resonance (NMR) spectroscopy. Enhancement of pi-stacking by quadrupolar interactions did not appear to play a significant role in stabilizing the conformations of heteropeptoids with alternating fluorinated and non-fluorinated side chains. However, incorporation of (S)-N-(1 (pentafluorophenyl)ethyl)glycine monomers enforced helicity in peptoids that typically exhibit threaded loop conformations. Moreover, we found that the incorporation of a single (S)-N-(1-(pentafluorophenyl)ethyl)glycine monomer could be used to selectively promote looped or helical structure in this important peptoid class by tuning the electronics of nearby heteroatoms. The strategic installation of this monomer unit represents a new approach for the manipulation of canonical peptoid structure and the construction of novel peptoid architectures. PMID- 17076513 TI - Carbene vs olefin products of C-H activation on ruthenium via competing alpha- and beta-H elimination. AB - Bulky pincer complexes of ruthenium are capable of C-H activation and H elimination from the pincer ligand backbone to produce mixtures of olefin and carbene products. To characterize the products and determine the mechanisms of the C-H cleavage, reactions of [RuCl(2)(p-cymene)](2) with N,N'-bis(di-tert butylphosphino)-1,3-diaminopropane (L1) and 1,3-bis(di-tert butylphosphinomethyl)cyclohexane (L2) were studied using a combination of X-ray crystallography, NMR spectroscopy, and DFT computational techniques. The reaction of L1 afforded a mixture of an alkylidene, a Fischer carbene, and two olefin isomers of the 16-e monohydride RuHCl[(t)Bu(2)PNHC(3)H(4)NHPBu(t)(2)] (2), whereas the reaction of L2 gave two olefin and two alkylidene isomers of 16-e RuHCl[2,6-(CH(2)PBu(t)(2))(2)C(6)H(8)] (3), all resulting from dehydrogenations of the ligand backbone of L1 and L2. The key intermediates implicated in the C-H activation reactions were identified as 14-electron paramagnetic species RuCl(PCP), where PCP = cyclometalated L1 or L2. Thus the alpha- and beta-H elimination reactions of RuCl(PCP) involved spin change and were formally spin forbidden. Hydrogenation of 2 and 3 afforded 16-electron dihydrides RuH(2)Cl(PCP) distinguished by a large quantum exchange coupling between the hydrides. PMID- 17076514 TI - Rotational reorganization of doped cholesteric liquid crystalline films. AB - In this paper an unprecedented rotational reorganization of cholesteric liquid crystalline films is described. This rotational reorganization results from the conversion of a chiral molecular motor dopant to an isomer with a different helical twisting power, leading to a change in the cholesteric pitch. The direction of this reorganization is correlated to the sign of the change in helical twisting power of the dopant. The rotational reorganization of the liquid crystalline film was used to rotate microscopic objects 4 orders of magnitude larger than the bistable dopants in the film, which shows that molecular motors and switches can perform work. The surface of the doped cholesteric liquid crystalline films was found to possess a regular surface relief, whose periodicity coincides with typical cholesteric polygonal line textures. These surface features originate from the cholesteric superstructure in the liquid crystalline film, which in turn is the result of the presence of the chiral dopant. As such, the presence of the dopant is expressed in these distinct surface structures. A possible mechanism at the origin of the rotational reorganization of liquid crystalline films and the cholesteric surface relief is discussed. PMID- 17076515 TI - Temperature-programmed microwave-assisted synthesis of SBA-15 ordered mesoporous silica. AB - The currently available microwave technology permits the development and implementation of a temperature-programmed microwave-assisted synthesis (TPMS) of ordered mesoporous silicas (OMSs). Unlike in previously reported syntheses of OMSs, in which only the final hydrothermal treatment was carried out under microwave irradiation, this work takes advantage of the existing capabilities of modern microwave systems to program the temperature and time for the entire synthesis of these materials. To demonstrate the flexibility of the proposed microwave-assisted synthesis, besides programming two consecutive steps involving initial stirring of the gel at a lower temperature and static hydrothermal treatment at a higher temperature, we explored the possibility of temperature programming of the latter step. A major advantage of microwave technology is the feasibility of temperature and time programming, which has been demonstrated by the synthesis of one of the most popular OMSs, SBA-15, over an unprecedented range of temperatures from 40 to 200 degrees C. Since the synthesis of OMSs has not yet been explored and reported at temperatures exceeding 150 degrees C, this work is focused on the SBA-15 samples prepared at higher temperatures (such as 160, 180, and even 200 degrees C). These SBA-15 samples show better thermal stability than those synthesized at commonly used temperatures either under conventional or microwave conditions. Moreover, a partial decomposition of the template during high-temperature microwave-assisted syntheses does not compromise the formation of well-ordered SBA-15 materials. This study shows that the simplicity and capability of temperature and time programming in TPMS allows one not only to tune the adsorption and structural properties of OMSs but also to easily screen a wide range of conditions in order to optimize and scale-up their preparation as well as to significantly reduce the time of synthesis from days to hours. PMID- 17076516 TI - Comparison of glass and plastic blood collection tubes. PMID- 17076518 TI - False-negative urine immunoassay after lorazepam overdose. PMID- 17076519 TI - Narrowing the differential diagnosis in surgical neuropathology. PMID- 17076520 TI - Opportunity knocks: the pathologist as laboratory genetics consultant. PMID- 17076521 TI - Effect of fondaparinux on coagulation assays: results of College of American Pathologists proficiency testing. AB - CONTEXT: Fondaparinux, a factor Xa inhibitor, is approved for thromboprophylaxis after orthopedic surgery and for treatment of venous thromboembolism. It may also be efficacious, safe, and cost-effective for other patients; thus, more widespread use of fondaparinux is likely. The effect of fondaparinux on coagulation testing needs to be thoroughly examined. OBJECTIVE: To report the effects of fondaparinux on coagulation tests (prothrombin time, activated partial thromboplastin time, fibrinogen, antithrombin, factor VIII, thrombin time, anti factor Xa) across diverse methodologies. DESIGN: Samples with different concentrations of fondaparinux (0, 0.4, 0.8, and 2.0 microg/mL) were sent to laboratories participating in the College of American Pathologists Comprehensive Coagulation proficiency survey (N = 898). Laboratory-specific methods were used to assay coagulation parameters. RESULTS: Prophylactic or therapeutic fondaparinux prolonged the prothrombin time by approximately 1 second and the activated partial thromboplastin time by 4 to 5 seconds, and reduced factor VIII from 119% to 107% and 102%, respectively. Supratherapeutic fondaparinux reduced factor VIII to 85%. The activated partial thromboplastin time was prolonged in 19%, 29%, and 52% of laboratories with prophylactic, therapeutic, and supratherapeutic fondaparinux levels, respectively. Fibrinogen, antithrombin, and thrombin time assays did not show clinically significant changes. When measuring fondaparinux concentration using an anti-factor Xa assay, the most accurate results were obtained when fondaparinux was used as the calibrator. CONCLUSIONS: Fondaparinux, even in prophylactic doses, slightly prolongs the prothrombin time and activated partial thromboplastin time and can interfere with factor VIII assays, but it has no clinically relevant effect on fibrinogen, antithrombin, or thrombin time. A fondaparinux standard curve should be used for reporting fondaparinux levels using an anti-factor Xa assay. PMID- 17076522 TI - Distinguishing carcinoid tumor of the mediastinum from thymoma: correlating cytologic features and performance in the College of American Pathologists Interlaboratory Comparison Program in Nongynecologic Cytopathology. AB - CONTEXT: The cytologic features of carcinoid tumor in mediastinal fine-needle aspiration are well described. Nevertheless, this tumor may be difficult to distinguish from thymoma in this site. OBJECTIVE: We sought to correlate the cytologic features of carcinoid tumor of the mediastinum in the College of American Pathologists Interlaboratory Comparison Program in Nongynecologic Cytopathology with the frequency of misclassification as thymoma. DESIGN: We reviewed 446 interpretations from 18 different cases of carcinoid tumor in mediastinum and correlated the cytologic features with performance. RESULTS: Cases were more frequently classified as thymoma (158 responses, 35%) than as carcinoid tumor (126 responses, 28%). The best-performing case was classified as carcinoid tumor only 56% of the time. Three cytologic patterns were identified. Four cases consisted of isolated round cells with salt-and-pepper chromatin. Four cases consisted of isolated spindle and round cells with salt-and-pepper chromatin. The remaining 10 cases consisted of cohesive fragments of crowded cells with finely granular chromatin showing numerous pyknotic cells mimicking lymphocytes. Prominent vasculature patterns were not a feature of any of the cases. There was no correlation between any pattern and the rate of classification as carcinoid tumor or thymoma (P > .05). CONCLUSIONS: Carcinoid tumor of the mediastinum is frequently misclassified as thymoma in this program. Although some cytologic patterns resemble thymoma, the lack of correlation of these patterns with performance suggests that at least part of the reason for misclassification may be failure to consider the correct diagnosis or a lack of familiarity with discriminating cytologic criteria. PMID- 17076523 TI - Protocol for the examination of specimens from patients with soft tissue tumors of intermediate malignant potential, malignant soft tissue tumors, and benign/locally aggressive and malignant bone tumors. PMID- 17076524 TI - An algorithmic approach to the brain biopsy--part I. AB - CONTEXT: The formulation of appropriate differential diagnoses for a slide is essential to the practice of surgical pathology but can be particularly challenging for residents and fellows. Algorithmic flow charts can help the less experienced pathologist to systematically consider all possible choices and eliminate incorrect diagnoses. They can assist pathologists-in-training in developing orderly, sequential, and logical thinking skills when confronting difficult cases. OBJECTIVE: To present an algorithmic flow chart as an approach to formulating differential diagnoses for lesions seen in surgical neuropathology. DESIGN: An algorithmic flow chart to be used in teaching residents. RESULTS: Algorithms are not intended to be final diagnostic answers on any given case. Algorithms do not substitute for training received from experienced mentors nor do they substitute for comprehensive reading by trainees of reference textbooks. Algorithmic flow diagrams can, however, direct the viewer to the correct spot in reference texts for further in-depth reading once they hone down their diagnostic choices to a smaller number of entities. The best feature of algorithms is that they remind the user to consider all possibilities on each case, even if they can be quickly eliminated from further consideration. CONCLUSIONS: In Part I, we assist the resident in learning how to handle brain biopsies in general and how to distinguish nonneoplastic lesions that mimic tumors from true neoplasms. PMID- 17076525 TI - An algorithmic approach to the brain biopsy--part II. AB - CONTEXT: The formulation of appropriate differential diagnoses for a slide is essential to the practice of surgical pathology but can be particularly challenging for residents and fellows. Algorithmic flow charts can help the less experienced pathologist to systematically consider all possible choices and eliminate incorrect diagnoses. They can assist pathologists-in-training in developing orderly, sequential, and logical thinking skills when confronting difficult cases. OBJECTIVE: To present an algorithmic flow chart as an approach to formulating differential diagnoses for lesions seen in surgical neuropathology. DESIGN: An algorithmic flow chart to be used in teaching residents. RESULTS: Algorithms are not intended to be final diagnostic answers on any given case. Algorithms do not substitute for training received from experienced mentors nor do they substitute for comprehensive reading by trainees of reference textbooks. Algorithmic flow diagrams can, however, direct the viewer to the correct spot in reference texts for further in-depth reading once they hone down their diagnostic choices to a smaller number of entities. The best feature of algorithms is that they remind the user to consider all possibilities on each case, even if they can be quickly eliminated from further consideration. CONCLUSIONS: In Part II, we assist the resident in arriving at the correct diagnosis for neuropathologic lesions containing granulomatous inflammation, macrophages, or abnormal blood vessels. PMID- 17076526 TI - The impact of the World Health Organization classification and clonality assessment of posttransplant lymphoproliferative disorders on disease management. AB - CONTEXT: The World Health Organization classification of posttransplant lymphoproliferative disorders divides them into 4 main categories. OBJECTIVE: To classify cases of posttransplant lymphoproliferative disorders diagnosed in our institution according to the World Health Organization scheme and correlate the classification and clonality with clinical data. DESIGN: Cases of posttransplant lymphoproliferative disorders were reviewed. They were classified according to the World Health Organization scheme. Clonality was determined by flow cytometry and/or polymerase chain reaction. Patients' charts were reviewed. RESULTS: Thirty one cases were identified. Median age was 33 years. There were 19 cases of kidney, 8 cases of liver, and 4 cases of bone marrow transplant. Immunosuppression consisted of cyclosporin A and prednisone (N = 24) or FK506 and prednisone (N = 7). Twenty cases (63%) were World Health Organization type 3, 7 cases (23%) type 2, 3 cases (6.4%) type 1, and 1 case type 4 posttransplant lymphoproliferative disorder. Ten patients received chemotherapy, 20 patients had reduction of immunosuppression, and 1 had no treatment. Follow-up was available on 25 patients. Seven (43.75%) of 16 with type 3 lesions with available follow-up died of their disease. Five of these patients received reduction of immunosuppression alone. Only 2 of 9 patients with type 3 disease who received chemotherapy died of disease. Two patients with type 2 disease died of unrelated causes. One patient is alive with disease; the remaining patients with types 1 and 2 disease are alive with no disease. CONCLUSIONS: The World Health Organization classification of posttransplant lymphoproliferative disorders is valuable in the identification of subtypes. It helps identify early lesions (1 and 2) requiring reduction of immunosuppression and type 3 disease, which requires chemotherapy from the outset. PMID- 17076527 TI - Prognostic significance of histomorphologic parameters in diffuse malignant peritoneal mesothelioma. AB - CONTEXT: Diffuse malignant peritoneal mesothelioma is currently regarded as a rare and lethal primary tumor arising from the peritoneal membrane. In the past, treatment plans with variable combinations of surgery and systemic chemotherapy were associated with a median survival of approximately 1 year. Standardized treatments using cytoreductive surgery and perioperative intraperitoneal chemotherapy have extended this survival. OBJECTIVE: To critically analyze the prognostic significance of histomorphologic parameters as a determinant of survival. DESIGN: Sixty-two consecutive patients with diffuse malignant peritoneal mesothelioma in whom data were prospectively accumulated were retrospectively analyzed by 14 different histomorphologic parameters. The influences of these pathologic characteristics on survival were critically statistically evaluated. RESULTS: In a univariate analysis, histologic type, nuclear/nucleolar size, stroma, depth of invasion into the bowel, atypical mitoses, mitotic index, necrosis, lymph node involvement, and chromatin patterns were found to be significant. In the multivariate analysis, histologic type and nuclear/nucleolar size remained as determinant histomorphologic characteristics. Use of biphasic/sarcomatoid histologic type as a poor prognostic characteristic was limited in that only 8% of patients showed this histology. CONCLUSIONS: Histomorphologic parameters carry prognostic significance in predicting the survival of patients with diffuse malignant peritoneal mesothelioma when treated in a standardized fashion using cytoreductive surgery and perioperative intraperitoneal chemotherapy. Nuclear/nucleolar size was found to be a reliable histomorphologic assessment available to assess prognosis in these patients. PMID- 17076528 TI - Patient safety in the clinical laboratory: a longitudinal analysis of specimen identification errors. AB - CONTEXT: Patient safety is an increasingly visible and important mission for clinical laboratories. Attention to improving processes related to patient identification and specimen labeling is being paid by accreditation and regulatory organizations because errors in these areas that jeopardize patient safety are common and avoidable through improvement in the total testing process. OBJECTIVE: To assess patient identification and specimen labeling improvement after multiple implementation projects using longitudinal statistical tools. DESIGN: Specimen errors were categorized by a multidisciplinary health care team. Patient identification errors were grouped into 3 categories: (1) specimen/requisition mismatch, (2) unlabeled specimens, and (3) mislabeled specimens. Specimens with these types of identification errors were compared preimplementation and postimplementation for 3 patient safety projects: (1) reorganization of phlebotomy (4 months); (2) introduction of an electronic event reporting system (10 months); and (3) activation of an automated processing system (14 months) for a 24-month period, using trend analysis and Student t test statistics. RESULTS: Of 16,632 total specimen errors, mislabeled specimens, requisition mismatches, and unlabeled specimens represented 1.0%, 6.3%, and 4.6% of errors, respectively. Student t test showed a significant decrease in the most serious error, mislabeled specimens (P < .001) when compared to before implementation of the 3 patient safety projects. Trend analysis demonstrated decreases in all 3 error types for 26 months. CONCLUSIONS: Applying performance improvement strategies that focus longitudinally on specimen labeling errors can significantly reduce errors, therefore improving patient safety. This is an important area in which laboratory professionals, working in interdisciplinary teams, can improve safety and outcomes of care. PMID- 17076529 TI - Extensively necrotic retinoblastoma is associated with high-risk prognostic factors. AB - CONTEXT: Retinoblastoma is the most common malignant intraocular tumor in children. It has been shown that adjuvant therapy following enucleation in patients with high-risk histopathologic features significantly decreases the mortality. We describe the association of extensive necrosis of tumor and intraocular structures with 2 of the major risk factors: optic nerve invasion and choroidal invasion. This may alert the pathologist who makes the observation of extensive necrosis to carefully search for histologic features associated with adverse outcome. OBJECTIVE: To determine whether extensively necrotic retinoblastoma is associated with high-risk histologic prognostic factors for metastatic disease and patient survival. DESIGN: Retrospective case series. Forty three eyes of 43 patients with retinoblastoma who underwent enucleation between 1990 and 2001 were evaluated. Medical records, histopathology specimens, pathology reports, and clinical photographs were reviewed. Tumors were designated as exhibiting extensive necrosis if more than 95% of tumor cells and intraocular tissues were necrotic. The main outcome measure was the association of extensive tumor necrosis with 3 high-risk histopathologic features: extraocular extension, optic nerve invasion, or choroidal invasion. Metastatic disease, patient survival, and associations with pathologic findings were also analyzed. RESULTS: Optic nerve head invasion (P < .001), post-lamina-cribrosal invasion (P < .001), and choroidal invasion by tumor (P = .004) were observed more frequently in eyes with extensive necrosis compared with eyes without extensive necrosis. Two of the 11 patients with extensively necrotic intraocular retinoblastoma died from metastatic disease (P = .06). None of the 32 patients without extensive necrosis developed metastatic disease or died. CONCLUSIONS: Extensive ocular tissue and tumor necrosis is associated with histologic high-risk prognostic factors for tumor metastasis and mortality. PMID- 17076530 TI - Aggressive clear cell chondrosarcomas: do distinctive characteristics exist?: a report of 4 cases. AB - CONTEXT: Clear cell chondrosarcoma (CCC) is commonly considered to be a low-grade subtype of chondrosarcoma. However, a few cases of CCC behave as high-grade lesions (with early metastases or multiple/synchronous locations). OBJECTIVE: To investigate morphologic features that can help predict the aggressiveness of these CCCs. DESIGN: To investigate possible hallmarks of this aggressiveness, we are presenting the clinicopathologic features of 6 cases of CCC, 4 of which presented aggressive features and 2 low-grade behavior. The patients were 5 men and 1 woman; their ages ranged from 22 to 47 years. Histologic appearance, ultrastructure, and immunohistochemical expression of metalloproteinase 1 and 2 and their inhibitors were evaluated in all 6 cases. RESULTS: Pain was the most common symptom; the lesions were located in the femur (4), humerus (2), and vertebral body (1), with 1 patient presenting a double/synchronous lesion. Although no major differences were detected using conventional light microscopy, an ultrastructural analysis--at variance with usual cases--showed a lack of superficial microvilli in more than 50% of neoplastic cells in the aggressive cases, therefore suggesting a less differentiated phenotype. In addition, metalloproteinase 2 was more diffusely expressed in the aggressive tumors than in the conventional CCCs, whereas p53 labeling was always negative. CONCLUSIONS: The aggressive behavior of some CCCs may be, at least in part, correlated to a lesser degree of cell differentiation and to the expression of tumor cell proteins, such as metalloproteinase 2, which are able to favor neoplastic spreading. PMID- 17076531 TI - Molecular detection of transfusion transmitted virus coinfection with some hepatotropic viruses. AB - CONTEXT: A novel human DNA virus was isolated from the serum of a patient with posttransfusion hepatitis; it was named transfusion transmitted virus (TTV). OBJECTIVE: To ascertain the influence of TTV (detected by polymerase chain reaction amplification of a conserved region of the viral genome) coinfection in individuals infected with hepatitis viruses (A, B, and C) and to investigate the putative role played by TTV in hepatic dysfunction in individuals with acute non A-E hepatitis. DESIGN: Sixty-two patients with viral hepatitis were included in the study in addition to 18 blood donors. Viral study of 4 hepatotropic viruses (A, B, C, and E) was carried out. Study for TTV DNA was performed by nested polymerase chain reaction. RESULTS: The prevalence of TTV was not statistically different between hepatitis patients and blood donors, and it was not correlated to the levels of the hepatic aspartate aminotransferase and alanine aminotransferase between individuals evidencing dual infection with hepatitis B and C viruses and healthy blood donors. However, in the group of patients with viral hepatitis of unknown etiology (non-A-E), those evidencing TTV viremia had statistically significant lower levels of alanine aminotransferase (P = .03) and aspartate aminotransferase (P = .04) than those who were TTV negative. CONCLUSIONS: We can conclude that TTV is a frequent virus isolated from patients with various types of viral hepatitis, from cases of hepatitis without obvious viral agent, and from the healthy population. TTV has no effect on biochemical markers of associated viral hepatitis. It may be associated with a mild form of non-A-E hepatitis. PMID- 17076532 TI - Specificity and sensitivity of hemosiderin-laden macrophages in routine bronchoalveolar lavage in children. AB - CONTEXT: The presence of iron or hemosiderin in macrophages obtained in routine bronchoalveolar lavage is considered crucial in the diagnosis of the clinical syndrome of hemosiderosis. However, there do not appear to be any data on the sensitivity and specificity of the finding of hemosiderin-laden macrophages (HLMs) in bronchoalveolar lavage in children. OBJECTIVE: To review data from bronchoalveolar lavage studies done in children to correlate the presence of HLMs with pneumonia and hemosiderosis and to determine what proportion of HLMs has the optimal sensitivity and specificity for the diagnosis of hemosiderosis. DESIGN: One hundred ten bronchoalveolar lavage specimens obtained via flexible bronchoscopy were reviewed retrospectively. The data collected for demographics, indication for the bronchoscopy, diagnosis of pneumonia, anemia, and bronchoscopy and bronchoalveolar lavage findings were compared between patients diagnosed with hemosiderosis and those diagnosed with other diseases. RESULTS: Six patients were diagnosed with hemosiderosis by clinical findings, lung biopsy, or autopsy. There were no statistical differences in pneumonia (P > .99), anemia (P > .99), or coughing (P = .08) between patients with hemosiderosis and other patients. Hemoptysis was the only symptom that was significantly different between the 2 groups (P = .04). The mean HLM index for patients with hemosiderosis was 56% +/- 16.17% and for other patients, 7.5% +/- 10.74% (P < .001). A HLM index of 35% gave a sensitivity of 1% and a specificity of .96%. CONCLUSIONS: These results confirm a strong association between HLM index and diagnosis of hemosiderosis in a pediatric population. Availability of this HLM index will result in accurate and timely diagnosis of pulmonary hemosiderosis, which may influence treatment and long-term prognosis. PMID- 17076533 TI - Nucleophosmin gene mutations in acute myeloid leukemia. AB - CONTEXT: Heterozygous mutation of the nucleophosmin gene (NPM1) has recently been described as one of the most frequent genetic lesions in acute myeloid leukemia (AML). OBJECTIVE: (1) To discuss the clinical, morphologic, immunophenotypic, and genetic features of AML with NPM1 gene mutations, along with various detection methods, (2) To explore the mechanisms by which NPM1 gene mutations contribute to leukemogenesis. DATA SOURCES/EXTRACTION: Data were analyzed from 7 recently published papers. RESULTS: NPM1 gene mutations tend to occur more frequently in women, and also tend to be associated with a higher white blood cell count. There is no significant age difference. NPM1-mutated AML is preferentially associated with AML with monocytic differentiation (in particular FAB M5b), lack of CD34, normal cytogenetics, FLT3 gene mutations, and a trend toward favorable clinical outcome, especially in patients without FLT3 gene mutation. NPM1 gene mutations cause a frame shift in the C-terminus of exon 12, disrupting the NPM nucleolar localization signal or generating a leucine-rich nuclear export motif, resulting in abnormal cytoplasmic accumulation of NPM. Several methods are suitable for detecting NPM1 gene mutation, including molecular and immunohistochemical studies. These mutations may contribute to leukemogenesis, at least in part, through disruption of the p14(ARF) (alternative reading frame) MDM2-p53 pathway and centrosomal duplication. CONCLUSIONS: Detection of NPM1 gene mutations may allow dissection of the heterogeneous group of AML with normal karyotype into prognostically different subgroups. Exploring the mechanisms may lead to a better understanding of how mutant NPM protein becomes leukemogenic, thereby providing insights for the development of new chemotherapeutic agents. PMID- 17076534 TI - Carcinoid tumors of the urinary tract and prostate. AB - CONTEXT: Carcinoid tumors are exceedingly rare in the genitourinary tract and may occur in the kidney, urinary bladder, urethra, or prostate. OBJECTIVE: To review the clinical and pathologic features of carcinoid tumors occurring in the urinary tract and prostate. DATA SOURCES: We searched the English language literature using MEDLINE and Ovid. CONCLUSIONS: Carcinoid tumors of the urinary tract and prostate share similar morphologic features with their counterparts in other organs. The differential diagnosis includes metastatic carcinoid tumor, paraganglioma, and nested variants of urothelial and prostatic carcinomas. Correlation of the clinical presentation and histopathologic features (including the immunohistochemical profile) will ensure accurate diagnosis of these rare tumors. PMID- 17076535 TI - Angioimmunoblastic T-cell lymphoma with supervening Epstein-Barr virus-associated large B-cell lymphoma. AB - Patients with angioimmunoblastic T-cell lymphoma can have profound immune dysfunction and immunodeficiency. Epstein-Barr virus-driven B-cell lymphoid proliferation can occur in angioimmunoblastic T-cell lymphoma, as in other immunodeficiency states. However, few cases of Epstein-Barr virus-positive B-cell lymphoma arising in patients with preexisting angioimmunoblastic T-cell lymphoma have been reported. We report a case of angioimmunoblastic T-cell lymphoma in which diffuse large B-cell lymphoma developed 56 months after the diagnosis of angioimmunoblastic T-cell lymphoma. The patient survived for 9 years after the initial diagnosis of angioimmunoblastic T-cell lymphoma, and molecular studies performed on multiple biopsy specimens during this period revealed the dynamic nature of clonal lymphoid expansion. Epstein-Barr virus latent membrane protein 1 and Epstein-Barr virus-encoded RNA were detected in the diffuse large B-cell lymphoma, suggesting that Epstein-Barr virus may have played a role in the pathogenesis of the diffuse large B-cell lymphoma. PMID- 17076536 TI - Large natural killer cell lymphoma arising from an indolent natural killer cell large granular lymphocyte proliferation. AB - Natural killer cell large granular lymphocyte proliferation is a relatively rare disorder that typically runs a chronic, indolent course. We present a patient with a 3 1/2-year history of natural killer cell large granular lymphocyte proliferation terminating in large cell lymphoma with natural killer cell features. The diagnosis of natural killer cell large granular lymphocyte proliferation was based on flow cytometric demonstration of an expanded population of CD3- CD16+/CD56+ lymphocytes in the peripheral blood. The patient experienced various rheumatologic symptoms, but was hematologically stable for 3 1/2 years. He then developed fevers, night sweats, weight loss, and a left lower lobe lung mass. Resection of the mass showed a large cell lymphoma with immunohistochemical positivity for CD2, CD7, CD56, and T-cell intracellular antigen-1, compatible with natural killer cell origin. In situ hybridization for Epstein-Barr virus and polymerase chain reaction analysis for T-cell receptor gene rearrangement were negative. To our knowledge, this is the second documented report of chronic natural killer cell large granular lymphocyte proliferation terminating in an aggressive large natural killer cell lymphoma. PMID- 17076537 TI - Paratesticular mucinous cystadenoma arising from an oviduct-like mullerian remnant: a case report and review of the literature. AB - A mucinous cystadenoma of the paratestis arising from an unequivocal oviduct-like structure in an 18-year-old man is reported. Mucinous cystadenomas of the testis and paratestis are extremely rare benign tumors of controversial origin. The diagnosis may be challenging and is of clinical importance because these lesions may mimic a teratoma, and teratomas in postpubertal males carry a malignant potential regardless of the degree of maturation. A few case reports and reviews of testicular and paratesticular mucinous neoplasms resembling ovarian tumors have suggested the possibility of a mullerian origin of these tumors. However, no histologic evidence has been found to support such a histogenesis. The current case demonstrates a clear transition from mullerian-type tubal epithelium to intestinal-type mucinous epithelium. This finding, in conjunction with immunophenotypic studies, strongly supports a derivation from a paratesticular mullerian remnant. To our knowledge this represents the first substantial evidence that, in at least some cases, mucinous cystadenomas of the testis and paratestis are of mullerian derivation. PMID- 17076538 TI - Follicular dendritic cell sarcoma arising in the dura mater of the spine. AB - There have been some individual case reports and a few large series of reports describing the clinicopathologic features of follicular dendritic cell sarcoma. This tumor originates from the dendritic cells of lymphoid follicles and is extremely rare in the extranodal location. We report the case of a 68-year-old man who presented with low back pain. A mass was found in the lumbar dura mater and extended to the right epidural space. The tumor was composed of nodules, sheets, and interlacing fascicles of oval-to-spindle cells intermingled with the dense infiltrates of small lymphocytes. Immunohistochemical staining revealed that the tumor cells were positive for CD21, CD23, CD35, and clusterin, and focally positive for CD68, CD20, and CD79a. To our knowledge, this is the first case of follicular dendritic cell sarcoma occurring in the dura mater of the spine. PMID- 17076539 TI - Vascular adrenal cysts: a brief review of the literature. AB - Adrenal cysts are rare and form a heterogeneous group of lesions that includes (a) parasitic cysts, (b) epithelial cysts, (c) pseudocysts, and (d) endothelial cysts. There is evidence (immunohistochemical and ultrastructural) that both pseudocysts and endothelial cysts are variants of vascular cysts. Adrenal vascular cysts account for 84% of adrenal cysts. They are more common in women and present clinically with abdominal pain or are incidental findings. Their imaging features are not specific. Grossly, both types of adrenal vascular cysts are encapsulated. Pseudocysts are unilocular, thick-walled, and devoid of endothelial lining, whereas endothelial cysts are thin-walled, multilocular, and lined by endothelium. Adrenal vascular cysts probably originate from a preexisting vascular hamartoma. The treatment of choice is surgical excision. The prognosis is excellent. PMID- 17076540 TI - An in-depth look at Krukenberg tumor: an overview. AB - Krukenberg tumor is an uncommon metastatic tumor of the ovary. This article provides an overview of the major pathologic manifestations of Krukenberg tumor, patient characteristics, clinical and laboratory features of the disease, prognostic factors, and current knowledge about its pathogenesis. Pathologists have to be familiar with the diagnostic histopathologic features of the tumor and its principal differential diagnoses. Awareness of the diagnostic manifestations of the tumor leads to the correct diagnosis and prevents tumor misclassification, thus avoiding improper clinical management. The article also addresses the potential clinical utility of serum CA 125 in patients with Krukenberg tumors. Prognosis of Krukenberg tumor is still very poor but our review of the literature reveals several factors that appear to have an impact on survival. There is no established treatment for Krukenberg tumors. A national registry and prospective studies are needed to set a therapeutic approach for Krukenberg tumors in the hope of improving the survival rate. PMID- 17076542 TI - Fluence rate or cumulative dose? Vulnerability of larval northern pike (Esox lucius) to ultraviolet radiation. AB - Newly hatched larvae of northern pike were exposed in the laboratory to four fluence rates of ultraviolet radiation (UVR; 290-400 nm) over three different time periods, resulting in total doses ranging from 3.0 +/- 0.2 to 63.0 +/- 4.4 kJ.m(-2). Mortality and behavior of the larvae were followed for 8-12 days, and growth measured at the end of the experiment. Also, the principle of reciprocity that the UVR-induced mortality depends on the cumulative dose, independent of fluence rate-was tested. Fluence rates higher than 1480 +/- 150 mW.m(-2) caused mortality and growth retardation. The highest fluence rate (3040 +/- 210 mW.m( 2)) caused 100% mortality in 5 days. All fluence rates caused behavioral disorders, which led to death at fluence rates higher than 1480 mW.m(-2). Reciprocity failure occurred with the lowest and highest dose (550 +/- 45 and 3040 +/- 210 mW.m(-2), respectively). The results show that fluence rate is of primary importance when assessing the UVR-related risk. PMID- 17076543 TI - Pressure-induced isomerization of retinal on bacteriorhodopsin as disclosed by fast magic angle spinning NMR. AB - Bacteriorhodopsin (bR) is a retinal protein in purple membrane of Halobacterium salinarum, which functions as a light-driven proton pump. We have detected pressure-induced isomerization of retinal in bR by analyzing 15N cross polarization-magic angle spinning (CP-MAS) NMR spectra of [zeta-15N]Lys-labeled bR. In the 15N-NMR spectra, both all-trans and 13-cis retinal configurations have been observed in the Lys N(zeta) in protonated Schiff base at 148.0 and 155.0 ppm, respectively, at the MAS frequency of 4 kHz in the dark. When the MAS frequency was increased up to 12 kHz corresponding to the sample pressure of 63 bar, the 15N-NMR signals of [zeta-15N]Lys in Schiff base of retinal were broadened. On the other hand, other [zeta-15N]Lys did not show broadening. Subsequently, the increased signal intensity of [zeta-15N]Lys in Schiff base of 13-cis retinal at 155.0 ppm was observed when the MAS frequency was decreased from 12 to 4 kHz. These results showed that the equilibrium constant of [all trans-bR]/[13-cis-bR] in retinal decreased by the pressure of 63 bar. It was also revealed that the structural changes induced by the pressure occurred in the vicinity of retinal. Therefore, microscopically, hydrogen-bond network around retinal would be disrupted or distorted by a constantly applied pressure. It is, therefore, clearly demonstrated that increased pressure induced by fast MAS frequencies generated isomerization of retinal from all-trans to 13-cis state in the membrane protein bR. PMID- 17076544 TI - Rapid chlorophyll a fluorescence transients of Lemna minor leaves as indication of light and exogenous electron carriers effect on photosystem II activity. AB - By using saturating flash, we investigated the change in the rapid fluorescence rise when Lemna minor leaf was exposed to different light conditions and treated with exogenous electron acceptors (methyl viologen and duroquinone) and electron donor (hydroxylamine). Investigation was carried out by using combined pulse amplitude modulated fluorometer and plant efficiency analyzer system, which were employed simultaneously to provide different light conditions and to induce rapid fluorescence rise respectively. We have shown that when leaf of L. minor was exposed to different conditions of illumination, rapid fluorescence rise was greatly influenced by the electron transport functions beyond quinone A plastoquinone reduction. This was indicated by the change in both fluorescence yield and appearance time of the different transients. When exogenous electron donor (hydroxylamine) and acceptors (methyl viologen and duroquinone) were applied in in vivo condition, we showed that rapid fluorescence rise represented a reliable indicator of PSII-PSI electron transport state and energy dissipation process. PMID- 17076545 TI - Why should medical students care about health policy? PMID- 17076546 TI - Anthropology in the clinic: the problem of cultural competency and how to fix it. PMID- 17076548 TI - Is it ethical for patients with renal disease to purchase kidneys from the world's poor? AB - BACKGROUND TO THE DEBATE: In many countries, the number of patients waiting for a kidney transplant is increasing. But there is a widespread and serious shortage of kidneys for transplantation, a shortage that can lead to suffering and death. One approach to tackling the shortage is for a patient with renal disease to buy a kidney from a living donor, who is often in a developing country, a sale that could--in theory at least--help to lift the donor out of poverty. Such kidney sales are almost universally illegal. Proponents of kidney sales argue that since the practice is widespread, it would be safer to formally regulate it, and that society should respect people's autonomous control over their bodies. Critics express concern about the potential for exploitation and coercion of the poor, and about the psychological and physical after-effects on the donors of this illegal kidney trade. PMID- 17076547 TI - Insights into the management of emerging infections: regulating variant Creutzfeldt-Jakob disease transfusion risk in the UK and the US. AB - BACKGROUND: Variant Creutzfeldt-Jakob disease (vCJD) is a human prion disease caused by infection with the agent of bovine spongiform encephalopathy. After the recognition of vCJD in the UK in 1996, many nations implemented policies intended to reduce the hypothetical risk of transfusion transmission of vCJD. This was despite the fact that no cases of transfusion transmission had yet been identified. In December 2003, however, the first case of vCJD in a recipient of blood from a vCJD-infected donor was announced. The aim of this study is to ascertain and compare the factors that influenced the motivation for and the design of regulations to prevent transfusion transmission of vCJD in the UK and US prior to the recognition of this case. METHODS AND FINDINGS: A document search was conducted to identify US and UK governmental policy statements and guidance, transcripts (or minutes when transcripts were not available) of scientific advisory committee meetings, research articles, and editorials published in medical and scientific journals on the topic of vCJD and blood transfusion transmission between March 1996 and December 2003. In addition, 40 interviews were conducted with individuals familiar with the decision-making process and/or the science involved. All documents and transcripts were coded and analyzed according to the methods and principles of grounded theory. Data showed that while resulting policies were based on the available science, social and historical factors played a major role in the motivation for and the design of regulations to protect against transfusion transmission of vCJD. First, recent experience with and collective guilt resulting from the transfusion-transmitted epidemics of HIV/AIDS in both countries served as a major, historically specific impetus for such policies. This history was brought to bear both by hemophilia activists and those charged with regulating blood products in the US and UK. Second, local specificities, such as the recall of blood products for possible vCJD contamination in the UK, contributed to a greater sense of urgency and a speedier implementation of regulations in that country. Third, while the results of scientific studies played a prominent role in the construction of regulations in both nations, this role was shaped by existing social and professional networks. In the UK, early focus on a European study implicating B-lymphocytes as the carrier of prion infectivity in blood led to the introduction of a policy that requires universal leukoreduction of blood components. In the US, early focus on an American study highlighting the ability of plasma to serve as a reservoir of prion infectivity led the FDA and its advisory panel to eschew similar measures. CONCLUSIONS: The results of this study yield three important theoretical insights that pertain to the global management of emerging infectious diseases. First, because the perception and management of disease may be shaped by previous experience with disease, especially catastrophic experience, there is always the possibility for over-management of some possible routes of transmission and relative neglect of others. Second, local specificities within a given nation may influence the temporality of decision making, which in turn may influence the choice of disease management policies. Third, a preference for science-based risk management among nations will not necessarily lead to homogeneous policies. This is because the exposure to and interpretation of scientific results depends on the existing social and professional networks within a given nation. Together, these theoretical insights provide a framework for analyzing and anticipating potential conflicts in the international management of emerging infectious diseases. In addition, this study illustrates the utility of qualitative methods in investigating research questions that are difficult to assess through quantitative means. PMID- 17076549 TI - Time to regenerate: ecosystems and health promotion. PMID- 17076550 TI - Time to renal disease and end-stage renal disease in PROFILE: a multiethnic lupus cohort. AB - BACKGROUND: Renal involvement is a serious manifestation of systemic lupus erythematosus (SLE); it may portend a poor prognosis as it may lead to end-stage renal disease (ESRD). The purpose of this study was to determine the factors predicting the development of renal involvement and its progression to ESRD in a multi-ethnic SLE cohort (PROFILE). METHODS AND FINDINGS: PROFILE includes SLE patients from five different United States institutions. We examined at baseline the socioeconomic-demographic, clinical, and genetic variables associated with the development of renal involvement and its progression to ESRD by univariable and multivariable Cox proportional hazards regression analyses. Analyses of onset of renal involvement included only patients with renal involvement after SLE diagnosis (n = 229). Analyses of ESRD included all patients, regardless of whether renal involvement occurred before, at, or after SLE diagnosis (34 of 438 patients). In addition, we performed a multivariable logistic regression analysis of the variables associated with the development of renal involvement at any time during the course of SLE. In the time-dependent multivariable analysis, patients developing renal involvement were more likely to have more American College of Rheumatology criteria for SLE, and to be younger, hypertensive, and of African American or Hispanic (from Texas) ethnicity. Alternative regression models were consistent with these results. In addition to greater accrued disease damage (renal damage excluded), younger age, and Hispanic ethnicity (from Texas), homozygosity for the valine allele of FcgammaRIIIa (FCGR3A*GG) was a significant predictor of ESRD. Results from the multivariable logistic regression model that included all cases of renal involvement were consistent with those from the Cox model. CONCLUSIONS: Fcgamma receptor genotype is a risk factor for progression of renal disease to ESRD. Since the frequency distribution of FCGR3A alleles does not vary significantly among the ethnic groups studied, the additional factors underlying the ethnic disparities in renal disease progression remain to be elucidated. PMID- 17076551 TI - Dementia before death in ageing societies--the promise of prevention and the reality. AB - BACKGROUND: Dementia and severe cognitive impairment are very closely linked to ageing. The longer we live the more likely we are to suffer from these conditions. Given population increases in longevity it is important to understand not only risk and protective factors for dementia and severe cognitive impairment at given ages but also whether protection affects cumulative risk. This can be explored by examining the effect on cumulative risk by time of death of factors found consistently to reduce risk at particular ages, such as education and social status. METHODS AND FINDINGS: In this analysis we report the prevalence of dementia and severe cognitive impairment in the year before death in a large population sample. In the Medical Research Council Cognitive Function and Ageing Study (a 10-y population-based cohort study of individuals 65 and over in England and Wales), these prevalences have been estimated by age, sex, social class, and education. Differences have been explored using logistic regression. The overall prevalence of dementia at death was 30%. There was a strong increasing trend for dementia with age from 6% for those aged 65-69 y at time of death to 58% for those aged 95 y and above at time of death. Higher prevalences were seen for severe cognitive impairment, with similar patterns. People with higher education and social class had significantly reduced dementia and severe cognitive impairment before death, but the absolute difference was small (under 10%). CONCLUSIONS: Reducing risk for dementia at a given age will lead to further extension of life, thus cumulative risk (even in populations at lower risk for given ages) remains high. Ageing of populations is likely to result in an increase in the number of people dying with dementia and severe cognitive impairment even in the presence of preventative programmes. Policy development and research for dementia must address the needs of individuals who will continue to experience these conditions before death. PMID- 17076552 TI - How did social medicine evolve, and where is it heading? PMID- 17076553 TI - HLA Alleles Associated with Delayed Progression to AIDS Contribute Strongly to the Initial CD8(+) T Cell Response against HIV-1. AB - BACKGROUND: Very little is known about the immunodominance patterns of HIV-1 specific T cell responses during primary HIV-1 infection and the reasons for human lymphocyte antigen (HLA) modulation of disease progression. METHODS AND FINDINGS: In a cohort of 104 individuals with primary HIV-1 infection, we demonstrate that a subset of CD8(+) T cell epitopes within HIV-1 are consistently targeted early after infection, while other epitopes subsequently targeted through the same HLA class I alleles are rarely recognized. Certain HLA alleles consistently contributed more than others to the total virus-specific CD8(+) T cell response during primary infection, and also reduced the absolute magnitude of responses restricted by other alleles if coexpressed in the same individual, consistent with immunodomination. Furthermore, individual HLA class I alleles that have been associated with slower HIV-1 disease progression contributed strongly to the total HIV-1-specific CD8(+) T cell response during primary infection. CONCLUSIONS: These data demonstrate consistent immunodominance patterns of HIV-1-specific CD8(+) T cell responses during primary infection and provide a mechanistic explanation for the protective effect of specific HLA class I alleles on HIV-1 disease progression. PMID- 17076554 TI - Ethnic disparities in health: the public's role in working for equality. PMID- 17076555 TI - Health is still social: contemporary examples in the age of the genome. PMID- 17076556 TI - Indigenous health and socioeconomic status in India. AB - BACKGROUND: Systematic evidence on the patterns of health deprivation among indigenous peoples remains scant in developing countries. We investigate the inequalities in mortality and substance use between indigenous and non indigenous, and within indigenous, groups in India, with an aim to establishing the relative contribution of socioeconomic status in generating health inequalities. METHODS AND FINDINGS: Cross-sectional population-based data were obtained from the 1998-1999 Indian National Family Health Survey. Mortality, smoking, chewing tobacco use, and alcohol use were four separate binary outcomes in our analysis. Indigenous status in the context of India was operationalized through the Indian government category of scheduled tribes, or Adivasis, which refers to people living in tribal communities characterized by distinctive social, cultural, historical, and geographical circumstances.Indigenous groups experience excess mortality compared to non-indigenous groups, even after adjusting for economic standard of living (odds ratio 1.22; 95% confidence interval 1.13-1.30). They are also more likely to smoke and (especially) drink alcohol, but the prevalence of chewing tobacco is not substantially different between indigenous and non-indigenous groups. There are substantial health variations within indigenous groups, such that indigenous peoples in the bottom quintile of the indigenous-peoples-specific standard of living index have an odds ratio for mortality of 1.61 (95% confidence interval 1.33-1.95) compared to indigenous peoples in the top fifth of the wealth distribution. Smoking, drinking alcohol, and chewing tobacco also show graded associations with socioeconomic status within indigenous groups. CONCLUSIONS: Socioeconomic status differentials substantially account for the health inequalities between indigenous and non indigenous groups in India. However, a strong socioeconomic gradient in health is also evident within indigenous populations, reiterating the overall importance of socioeconomic status for reducing population-level health disparities, regardless of indigeneity. PMID- 17076557 TI - Analysis of gene expression using gene sets discriminates cancer patients with and without late radiation toxicity. AB - BACKGROUND: Radiation is an effective anti-cancer therapy but leads to severe late radiation toxicity in 5%-10% of patients. Assuming that genetic susceptibility impacts this risk, we hypothesized that the cellular response of normal tissue to X-rays could discriminate patients with and without late radiation toxicity. METHODS AND FINDINGS: Prostate carcinoma patients without evidence of cancer 2 y after curative radiotherapy were recruited in the study. Blood samples of 21 patients with severe late complications from radiation and 17 patients without symptoms were collected. Stimulated peripheral lymphocytes were mock-irradiated or irradiated with 2-Gy X-rays. The 24-h radiation response was analyzed by gene expression profiling and used for classification. Classification was performed either on the expression of separate genes or, to augment the classification power, on gene sets consisting of genes grouped together based on function or cellular colocalization.X-ray irradiation altered the expression of radio-responsive genes in both groups. This response was variable across individuals, and the expression of the most significant radio-responsive genes was unlinked to radiation toxicity. The classifier based on the radiation response of separate genes correctly classified 63% of the patients. The classifier based on affected gene sets improved correct classification to 86%, although on the individual level only 21/38 (55%) patients were classified with high certainty. The majority of the discriminative genes and gene sets belonged to the ubiquitin, apoptosis, and stress signaling networks. The apoptotic response appeared more pronounced in patients that did not develop toxicity. In an independent set of 12 patients, the toxicity status of eight was predicted correctly by the gene set classifier. CONCLUSIONS: Gene expression profiling succeeded to some extent in discriminating groups of patients with and without severe late radiotherapy toxicity. Moreover, the discriminative power was enhanced by assessment of functionally or structurally related gene sets. While prediction of individual response requires improvement, this study is a step forward in predicting susceptibility to late radiation toxicity. PMID- 17076558 TI - "Anecdotal evidence": why narratives matter to medical practice. PMID- 17076559 TI - Methods of blinding in reports of randomized controlled trials assessing pharmacologic treatments: a systematic review. AB - BACKGROUND: Blinding is a cornerstone of therapeutic evaluation because lack of blinding can bias treatment effect estimates. An inventory of the blinding methods would help trialists conduct high-quality clinical trials and readers appraise the quality of results of published trials. We aimed to systematically classify and describe methods to establish and maintain blinding of patients and health care providers and methods to obtain blinding of outcome assessors in randomized controlled trials of pharmacologic treatments. METHODS AND FINDINGS: We undertook a systematic review of all reports of randomized controlled trials assessing pharmacologic treatments with blinding published in 2004 in high impact factor journals from Medline and the Cochrane Methodology Register. We used a standardized data collection form to extract data. The blinding methods were classified according to whether they primarily (1) established blinding of patients or health care providers, (2) maintained the blinding of patients or health care providers, and (3) obtained blinding of assessors of the main outcomes. We identified 819 articles, with 472 (58%) describing the method of blinding. Methods to establish blinding of patients and/or health care providers concerned mainly treatments provided in identical form, specific methods to mask some characteristics of the treatments (e.g., added flavor or opaque coverage), or use of double dummy procedures or simulation of an injection. Methods to avoid unblinding of patients and/or health care providers involved use of active placebo, centralized assessment of side effects, patients informed only in part about the potential side effects of each treatment, centralized adapted dosage, or provision of sham results of complementary investigations. The methods reported for blinding outcome assessors mainly relied on a centralized assessment of complementary investigations, clinical examination (i.e., use of video, audiotape, or photography), or adjudication of clinical events. CONCLUSIONS: This review classifies blinding methods and provides a detailed description of methods that could help trialists overcome some barriers to blinding in clinical trials and readers interpret the quality of pharmacologic trials. PMID- 17076560 TI - Can we prevent, delay, or shorten the course of dementia? PMID- 17076561 TI - Atypical haemolytic uraemic syndrome associated with a hybrid complement gene. AB - BACKGROUND: Sequence analysis of the regulators of complement activation (RCA) cluster of genes at chromosome position 1q32 shows evidence of several large genomic duplications. These duplications have resulted in a high degree of sequence identity between the gene for factor H (CFH) and the genes for the five factor H-related proteins (CFHL1-5; aliases CFHR1-5). CFH mutations have been described in association with atypical haemolytic uraemic syndrome (aHUS). The majority of the mutations are missense changes that cluster in the C-terminal region and impair the ability of factor H to regulate surface-bound C3b. Some have arisen as a result of gene conversion between CFH and CFHL1. In this study we tested the hypothesis that nonallelic homologous recombination between low copy repeats in the RCA cluster could result in the formation of a hybrid CFH/CFHL1 gene that predisposes to the development of aHUS. METHODS AND FINDINGS: In a family with many cases of aHUS that segregate with the RCA cluster we used cDNA analysis, gene sequencing, and Southern blotting to show that affected individuals carry a heterozygous CFH/CFHL1 hybrid gene in which exons 1-21 are derived from CFH and exons 22/23 from CFHL1. This hybrid encodes a protein product identical to a functionally significant CFH mutant (c.3572C>T, S1191L and c.3590T>C, V1197A) that has been previously described in association with aHUS. CONCLUSIONS: CFH mutation screening is recommended in all aHUS patients prior to renal transplantation because of the high risk of disease recurrence post transplant in those known to have a CFH mutation. Because of our finding it will be necessary to implement additional screening strategies that will detect a hybrid CFH/CFHL1 gene. PMID- 17076562 TI - Complement factor h gene abnormalities in haemolytic uraemic syndrome: from point mutations to hybrid gene. PMID- 17076563 TI - HIV, stigma, and rates of infection: a rumour without evidence. PMID- 17076564 TI - Is there a global bioethics? End-of-life in Thailand and the case for local difference. PMID- 17076565 TI - Can the severity of normal tissue damage after radiation therapy be predicted? PMID- 17076567 TI - An ethnographic study of the social context of migrant health in the United States. AB - BACKGROUND: Migrant workers in the United States have extremely poor health. This paper aims to identify ways in which the social context of migrant farm workers affects their health and health care. METHODS AND FINDINGS: This qualitative study employs participant observation and interviews on farms and in clinics throughout 15 months of migration with a group of indigenous Triqui Mexicans in the western US and Mexico. Study participants include more than 130 farm workers and 30 clinicians. Data are analyzed utilizing grounded theory, accompanied by theories of structural violence, symbolic violence, and the clinical gaze. The study reveals that farm working and housing conditions are organized according to ethnicity and citizenship. This hierarchy determines health disparities, with undocumented indigenous Mexicans having the worst health. Yet, each group is understood to deserve its place in the hierarchy, migrant farm workers often being blamed for their own sicknesses. CONCLUSIONS: Structural racism and anti immigrant practices determine the poor working conditions, living conditions, and health of migrant workers. Subtle racism serves to reduce awareness of this social context for all involved, including clinicians. The paper concludes with strategies toward improving migrant health in four areas: health disparities research, clinical interactions with migrant laborers, medical education, and policy making. PMID- 17076568 TI - Structural violence and clinical medicine. PMID- 17076570 TI - Research ethics boards: size, not money. PMID- 17076569 TI - Reinterpreting ethnic patterns among white and African American men who inject heroin: a social science of medicine approach. AB - BACKGROUND: Street-based heroin injectors represent an especially vulnerable population group subject to negative health outcomes and social stigma. Effective clinical treatment and public health intervention for this population requires an understanding of their cultural environment and experiences. Social science theory and methods offer tools to understand the reasons for economic and ethnic disparities that cause individual suffering and stress at the institutional level. METHODS AND FINDINGS: We used a cross-methodological approach that incorporated quantitative, clinical, and ethnographic data collected by two contemporaneous long-term San Francisco studies, one epidemiological and one ethnographic, to explore the impact of ethnicity on street-based heroin-injecting men 45 years of age or older who were self-identified as either African American or white. We triangulated our ethnographic findings by statistically examining 14 relevant epidemiological variables stratified by median age and ethnicity. We observed significant differences in social practices between self-identified African Americans and whites in our ethnographic social network sample with respect to patterns of (1) drug consumption; (2) income generation; (3) social and institutional relationships; and (4) personal health and hygiene. African Americans and whites tended to experience different structural relationships to their shared condition of addiction and poverty. Specifically, this generation of San Francisco injectors grew up as the children of poor rural to urban immigrants in an era (the late 1960s through 1970s) when industrial jobs disappeared and heroin became fashionable. This was also when violent segregated inner city youth gangs proliferated and the federal government initiated its "War on Drugs." African Americans had earlier and more negative contact with law enforcement but maintained long-term ties with their extended families. Most of the whites were expelled from their families when they began engaging in drug-related crime. These historical-structural conditions generated distinct presentations of self. Whites styled themselves as outcasts, defeated by addiction. They professed to be injecting heroin to stave off "dopesickness" rather than to seek pleasure. African Americans, in contrast, cast their physical addiction as an oppositional pursuit of autonomy and pleasure. They considered themselves to be professional outlaws and rejected any appearance of abjection. Many, but not all, of these ethnographic findings were corroborated by our epidemiological data, highlighting the variability of behaviors within ethnic categories. CONCLUSIONS: Bringing quantitative and qualitative methodologies and perspectives into a collaborative dialog among cross-disciplinary researchers highlights the fact that clinical practice must go beyond simple racial or cultural categories. A clinical social science approach provides insights into how sociocultural processes are mediated by historically rooted and institutionally enforced power relations. Recognizing the logical underpinnings of ethnically specific behavioral patterns of street based injectors is the foundation for cultural competence and for successful clinical relationships. It reduces the risk of suboptimal medical care for an exceptionally vulnerable and challenging patient population. Social science approaches can also help explain larger-scale patterns of health disparities; inform new approaches to structural and institutional-level public health initiatives; and enable clinicians to take more leadership in changing public policies that have negative health consequences. PMID- 17076571 TI - Research ethics boards: error and misconception. PMID- 17076572 TI - Research ethics boards: no data on quality of for-profit or non-profit IRBs. PMID- 17076574 TI - Novel therapies for tuberculosis: tuberculosis control and its discontents. PMID- 17076575 TI - Preterm birth: effect of corticosteroids or immediate cord clamping? PMID- 17076576 TI - Industry-sponsored research: a more comprehensive alternative. PMID- 17076579 TI - Profiling of CSF: reliability of diagnosis. PMID- 17076580 TI - Profiling of CSF: small subgroups. PMID- 17076582 TI - Research ethics boards: the protection of human subjects. PMID- 17076583 TI - Mapping the economic costs and benefits of conservation. AB - Resources for biodiversity conservation are severely limited, requiring strategic investment. Understanding both the economic benefits and costs of conserving ecosystems will help to allocate scarce dollars most efficiently. However, although cost-benefit analyses are common in many areas of policy, they are not typically used in conservation planning. We conducted a spatial evaluation of the costs and benefits of conservation for a landscape in the Atlantic forests of Paraguay. We considered five ecosystem services (i.e., sustainable bushmeat harvest, sustainable timber harvest, bioprospecting for pharmaceutical products, existence value, and carbon storage in aboveground biomass) and compared them to estimates of the opportunity costs of conservation. We found a high degree of spatial variability in both costs and benefits over this relatively small (approximately 3,000 km(2)) landscape. Benefits exceeded costs in some areas, with carbon storage dominating the ecosystem service values and swamping opportunity costs. Other benefits associated with conservation were more modest and exceeded costs only in protected areas and indigenous reserves. We used this cost-benefit information to show that one potential corridor between two large forest patches had net benefits that were three times greater than two otherwise similar alternatives. Spatial cost-benefit analysis can powerfully inform conservation planning, even though the availability of relevant data may be limited, as was the case in our study area. It can help us understand the synergies between biodiversity conservation and economic development when the two are indeed aligned and to clearly understand the trade-offs when they are not. PMID- 17076584 TI - Human immunodeficiency virus impairs reverse cholesterol transport from macrophages. AB - Several steps of HIV-1 replication critically depend on cholesterol. HIV infection is associated with profound changes in lipid and lipoprotein metabolism and an increased risk of coronary artery disease. Whereas numerous studies have investigated the role of anti-HIV drugs in lipodystrophy and dyslipidemia, the effects of HIV infection on cellular cholesterol metabolism remain uncharacterized. Here, we demonstrate that HIV-1 impairs ATP-binding cassette transporter A1 (ABCA1)-dependent cholesterol efflux from human macrophages, a condition previously shown to be highly atherogenic. In HIV-1-infected cells, this effect was mediated by Nef. Transfection of murine macrophages with Nef impaired cholesterol efflux from these cells. At least two mechanisms were found to be responsible for this phenomenon: first, HIV infection and transfection with Nef induced post-transcriptional down-regulation of ABCA1; and second, Nef caused redistribution of ABCA1 to the plasma membrane and inhibited internalization of apolipoprotein A-I. Binding of Nef to ABCA1 was required for down-regulation and redistribution of ABCA1. HIV-infected and Nef-transfected macrophages accumulated substantial amounts of lipids, thus resembling foam cells. The contribution of HIV-infected macrophages to the pathogenesis of atherosclerosis was supported by the presence of HIV-positive foam cells in atherosclerotic plaques of HIV infected patients. Stimulation of cholesterol efflux from macrophages significantly reduced infectivity of the virions produced by these cells, and this effect correlated with a decreased amount of virion-associated cholesterol, suggesting that impairment of cholesterol efflux is essential to ensure proper cholesterol content in nascent HIV particles. These results reveal a previously unrecognized dysregulation of intracellular lipid metabolism in HIV-infected macrophages and identify Nef and ABCA1 as the key players responsible for this effect. Our findings have implications for pathogenesis of both HIV disease and atherosclerosis, because they reveal the role of cholesterol efflux impairment in HIV infectivity and suggest a possible mechanism by which HIV infection of macrophages may contribute to increased risk of atherosclerosis in HIV-infected patients. PMID- 17076585 TI - Phenotypic plasticity opposes species invasions by altering fitness surface. AB - Understanding species invasion is a central problem in ecology because invasions of exotic species severely impact ecosystems, and because invasions underlie fundamental ecological processes. However, the influence on invasions of phenotypic plasticity, a key component of many species interactions, is unknown. We present a model in which phenotypic plasticity of a resident species increases its ability to oppose invaders, and plasticity of an invader increases its ability to displace residents. Whereas these effects are expected due to increased fitness associated with phenotypic plasticity, the model additionally reveals a new and unforeseen mechanism by which plasticity affects invasions: phenotypic plasticity increases the steepness of the fitness surface, thereby making invasion more difficult, even by phenotypically plastic invaders. Our results should apply to phenotypically plastic responses to any fluctuating environmental factors including predation risk, and to other factors that affect the fitness surface such as the generalism of predators. We extend the results to competition, and argue that phenotypic plasticity's effect on the fitness surface will destabilize coexistence at local scales, but stabilize coexistence at regional scales. Our study emphasizes the need to incorporate variable interaction strengths due to phenotypic plasticity into invasion biology and ecological theory on competition and coexistence in fragmented landscapes. PMID- 17076586 TI - Conservation planning for ecosystem services. AB - Despite increasing attention to the human dimension of conservation projects, a rigorous, systematic methodology for planning for ecosystem services has not been developed. This is in part because flows of ecosystem services remain poorly characterized at local-to-regional scales, and their protection has not generally been made a priority. We used a spatially explicit conservation planning framework to explore the trade-offs and opportunities for aligning conservation goals for biodiversity with six ecosystem services (carbon storage, flood control, forage production, outdoor recreation, crop pollination, and water provision) in the Central Coast ecoregion of California, United States. We found weak positive and some weak negative associations between the priority areas for biodiversity conservation and the flows of the six ecosystem services across the ecoregion. Excluding the two agriculture-focused services-crop pollination and forage production-eliminates all negative correlations. We compared the degree to which four contrasting conservation network designs protect biodiversity and the flow of the six services. We found that biodiversity conservation protects substantial collateral flows of services. Targeting ecosystem services directly can meet the multiple ecosystem services and biodiversity goals more efficiently but cannot substitute for targeted biodiversity protection (biodiversity losses of 44% relative to targeting biodiversity alone). Strategically targeting only biodiversity plus the four positively associated services offers much promise (relative biodiversity losses of 7%). Here we present an initial analytical framework for integrating biodiversity and ecosystem services in conservation planning and illustrate its application. We found that although there are important potential trade-offs between conservation for biodiversity and for ecosystem services, a systematic planning framework offers scope for identifying valuable synergies. PMID- 17076587 TI - Reactivity of conjugated and unconjugated pterins with singlet oxygen (O2(1Deltag)): physical quenching and chemical reaction. AB - Pterins (PTs) belong to a class of heterocyclic compounds present in a wide range of living systems. They participate in relevant biological functions and are involved in different photobiological processes. We have investigated the reactivity of conjugated PTs (folic acid [FA], 10-methylfolic acid [MFA], pteroic acid [PA]) and unconjugated PTs (PT, 6-hydroxymethylpterin [HPT], 6-methylpterin [MPT], 6,7-dimethylpterin [DPT], rhamnopterin [RPT]) with singlet oxygen (1O2) in aqueous solutions, and compared the efficiencies of chemical reaction and physical quenching. The chemical reactions between 1O2, produced by photosensitization, and PT derivatives were followed by UV-visible spectrophotometry and high-performance liquid chromatography, and corresponding rate constants (k(r)) were evaluated. Whenever possible, products were identified and quantified. Rate constants of 1O2 total quenching by the PT derivatives investigated were obtained from steady-state 1O2 luminescence measurements. Results show that the behavior of conjugated PTs differs considerably from that of unconjugated derivatives, and the mechanisms of 1O2 physical quenching by these compounds and of their chemical reaction with 1O2 are discussed in relation to their structural features. PMID- 17076588 TI - Aimless excursions: wandering in the elderly. AB - One in five people with dementia wander. Long-term care facilities must deal with wandering while simultaneously protecting residents, complying with life-safety codes, maintaining a home-like environment, and respecting resident rights. Wanderers tend to be physically healthier than those who do not wander, but more behaviorally disturbed. Experts indicate that 72% of patients who elope will attempt to elope again. Strategies like using commercial security systems, landscaping to direct wanderers' attention back to the facility, keeping resident albums with recent pictures and facts about the resident, and having a research based policy help prevent serious outcomes. PMID- 17076589 TI - Gender-specific pharmacology: implications for therapy. AB - Historically, research into drugs and diseases has assumed that beyond the reproductive system, gender differences did not exist or were not relevant. Interest in recognizing the importance of gender-based differences in pharmacology and disease has been fueled in recent years by an increasing amount of data revealing variations in drug efficacy and side effect profiles between the genders as well as differences in disease prevalence. PMID- 17076590 TI - Counter sampling combined with medical provider education: do they alter prescribing behavior? AB - OBJECTIVE: To observe if medical providers alter their prescribing patterns of three relatively expensive categories of medications provided as samples by manufacturers (focus medications) when they receive additional education from pharmacists concerning the appropriate use of lower cost alternatives (counter samples) that are made available to dispense. DESIGN: Pretest, post-test with a control group. SETTING: Two rural, private care clinics in southeastern Idaho providing immediate care services. PARTICIPANTS: Eight medical providers at a clinic where interventions were employed (active intervention group) and seven medical providers in a clinic where no interventions occurred (control group). INTERVENTIONS: Medical providers in the active intervention group had: 1) education from pharmacists concerning the appropriate use of lower-cost alternatives compared with expensive focus medications 2) counter samples and patient sample handouts available to dispense to patients at their own discretion. MAIN OUTCOME MEASURES: The percentage of the total yearly prescriptions for nonsteroidal anti-inflammatory drugs (NSAIDs), antihistamines, and acid-relief medications that consisted of focus-COX-2 NSAIDs, nonsedating antihistamines, and proton pump inhibitors (PPIs), respectively. RESULTS: The prescribing behavior of medical providers in the active intervention and control groups were significantly different at baseline in all three categories of focus medications. This suggested that the results should focus on changes across the two years of the study within the intervention and control groups rather than across the two groups. Medical providers in the intervention group significantly decreased the use of COX-2 NSAID prescriptions relative to total NSAID prescriptions following active intervention (38.9% in year 1 versus 23.7% in year 2, P < 0.05). Over the same two time periods, a nonstatistically significant decrease in COX-2 NSAID prescribing was seen at the control site (67.5% versus 62%, P > 0.05). Education and counter sampling did not stop medical providers from significantly increasing the total yearly prescriptions for antihistamines and acid-relief medications that consisted of focus-nonsedating antihistamines (86.7% versus 93.1%, P < 0.05) and PPIs (68.9% versus 86.2%, P < 0.05). Statistically significant increases in the prescribing of focus-nonsedating antihistamines (77.9% versus 98.3%, P < 0.05) and PPIs (77.5% versus 91.4%, P < 0.05) were also observed in the control group. CONCLUSIONS: Education by pharmacists, combined with access to counter samples, may or may not have an effect on medical provider prescribing, depending on the category of medication targeted for cost control. PMID- 17076591 TI - Resolving medication discrepancies. AB - When elderly patients transition from one health care setting to another they are at increased risk of experiencing fragmented care. One of the most common manifestations of this lack of continuity is medication discrepancy. These differences, such as omission of medications, dosing errors, and duplication of therapy, are a significant safety issue for seniors. This case study describes a 76-year-old woman who is a new resident in an assisted living facility and is referred to the consultant pharmacist for medication review and evaluation of possible adverse drug events. In reviewing the drug regimen, the pharmacist uncovered medication discrepancies; other members of the health care team were unaware of them. These discrepancies may have contributed to some of the patient's current health problems. This case study demonstrates the role of the consultant pharmacist during transitions of care setting and reviews strategies that the pharmacist can use to identify and resolve medication discrepancies. PMID- 17076592 TI - Clinical pearls: management of depression in the elderly. PMID- 17076593 TI - The CNS as a target for peptides and peptide-based drugs. AB - Peptides hold great potential as CNS drugs, but their delivery to the CNS is problematic. However, actual roadblocks to peptide delivery are different from those often perceived. Many peptides cross the blood-brain barrier by saturable and non-saturable mechanisms, and accumulate in brain in amounts sufficient to produce physiological effects. Peripheral factors (e.g., short half-life in blood) can be dominant factors limiting therapeutic use. Production of therapeutics that are enzymatically resistant and have long circulation times, even when the blood-brain barrier penetration is low, can result in substances with significant CNS accumulation. Surprisingly low amounts of peptide in brain can result in CNS effects, and so the dose needed for brain delivery is generally much smaller than for peripheral tissues. Brain-to-blood transporters can greatly limit CNS accumulation of a potential therapeutic. Finally, intranasal and intrathecal routes may be especially useful for substances that are rapidly degraded in blood or are large and hydrophobic, respectively. PMID- 17076594 TI - Ultrasound-induced cavitation: applications in drug and gene delivery. AB - Ultrasound, which has been conventionally used for diagnostics until recently, is now being extensively used for drug and gene delivery. This transformation has come about primarily due to ultrasound-mediated acoustic cavitation - particularly transient cavitation. Acoustic cavitation has been used to facilitate the delivery of small molecules, as well as macromolecules, including proteins and DNA. Controlled generation of cavitation has also been used for targeting drugs to diseased tissues, including skin, brain, eyes and endothelium. Ultrasound has also been employed for the treatment of several diseases, including thromboembolism, arteriosclerosis and cancer. This review provides a detailed account of mechanisms, current status and future prospects of ultrasonic cavitation in drug and gene delivery applications. PMID- 17076595 TI - Transfersomes for transdermal drug delivery. AB - Transfersomes (Idea AG) are a form of elastic or deformable vesicle, which were first introduced in the early 1990s. Elasticity is generated by incorporation of an edge activator in the lipid bilayer structure. The original composition of these vesicles was soya phosphatidyl choline incorporating sodium cholate and a small concentration of ethanol. Transfersomes are applied in a non-occluded method to the skin and have been shown to permeate through the stratum corneum lipid lamellar regions as a result of the hydration or osmotic force in the skin. They have been used as drug carriers for a range of small molecules, peptides, proteins and vaccines, both in vitro and in vivo. It has been claimed by Idea AG that intact Transfersomes penetrate through the stratum corneum and the underlying viable skin into the blood circulation. However, this has not been substantiated by other research groups who have extensively probed the mechanism of penetration and interaction of elastic vesicles in the skin. Structural changes in the stratum corneum have been identified, and intact elastic vesicles visualised within the stratum corneum lipid lamellar regions, but no intact vesicles have been ascertained in the viable tissues. Using the principle of incorporating an edge-activator agent into a bilayer structure, a number of other elastic vesicle compositions have been evaluated. This review describes the research into the development and evaluation of Transfersomes and elastic vesicles as topical and transdermal delivery systems. PMID- 17076596 TI - Influence of protein transduction domains on intracellular delivery of macromolecules. AB - As the plasma membrane and blood-brain barrier selectively restrict the entry of most compounds into cells to < 500 Da, delivering macromolecules into cells was, until recently, little more than a goal. However, with significant effort to capitalise on therapeutic targets available in the post-genomic era, novel approaches for delivering therapeutic macromolecules are being rapidly developed. The discovery of small cationic peptides, termed peptide/protein transduction domains or cell-penetrating peptides, which cross biological membranes, has emerged as a venerable Trojan horse to transport large, biologically active molecules, such as peptides, proteins and oligonucleotides, into mammalian cells in vitro, as well as in preclinical models and clinical trials in vivo. This review discusses the implications of peptide/protein transduction domain-mediated delivery of macromolecules and their possible uses as important primary drug delivery agents. PMID- 17076597 TI - Delivery systems and adjuvants for oral vaccines. AB - The oral route is the ideal means of delivering prophylactic and therapeutic vaccines, offering significant advantages over systemic delivery. Most notably, oral delivery is associated with simple administration and improved safety. In addition, unlike systemic immunisation, oral delivery can induce mucosal immune responses. However, the oral route of vaccine delivery is the most difficult because of the numerous barriers posed by the gastrointestinal tract. To facilitate effective immunisation with peptide and protein vaccines, antigens must be protected, uptake enhanced and the innate immune response activated. Numerous delivery systems and adjuvants have been evaluated for oral vaccine delivery, including live vectors, inert particles and bacterial toxins. Although developments in oral vaccines have been disappointing so far, in terms of the generation of products, the availability of a range of novel delivery systems offers much greater hope for the future development of improved oral vaccines. PMID- 17076598 TI - Non-invasive routes for insulin administration: current state and perspectives. AB - Diabetes mellitus is a chronic disease that usually requires multiple insulin injections to achieve adequate glycaemic control. This represents a major cause of reduced compliance to treatment. Consequently, other routes for insulin administration have been explored. During recent years, much progress in the development of inhaled insulin has been made. Inhaled insulin has shown favourable properties, such as a rapid onset of action, improved bioavailability and good tolerability; thereby providing satisfaction and ease of administration. However, long-term safety of inhaled insulin needs to be assessed, and the cost would be higher than injectable insulin. Nasal, oral and transdermal insulins are undergoing early phases of pharmacological development. The purpose of this review is to describe the latest developments in the area of non-invasive routes for insulin delivery. PMID- 17076599 TI - Delivery of lysosomal enzymes for therapeutic use: glucocerebrosidase as an example. AB - Enzyme therapies for lysosomal storage diseases have developed over the past decade into the standard-of-care for affected patients. Such therapy for Gaucher disease has been the prototype, using natural source or recombinant forms of human acid beta-glucosidase (GCase). In Gaucher disease, macrophages are the repository for the pathological lipid and the target for delivery of GCase. The macrophage mannose receptor provides a Trojan horse for intracellular delivery of intravenously administered GCase (man-GCase) with mannosyl-terminated oligosaccharide chains. Passage through several hostile compartments (e.g., plasma) leads to inefficient delivery of man-GCase to macrophage lysosomes. However, regular infusions of man-GCase re-establishes health in affected patients. Similar results are being obtained in several other lysosomal storage diseases. Evolving gene and chaperone approaches provide alternative treatment strategies. PMID- 17076600 TI - Progressive stent technologies: new approaches for the treatment of cardiovascular diseases. AB - The CYPHER (Cordis, Johnson & Johnson) sirolimus-eluting stent and the TAXU (Boston Scientific) paclitaxel-eluting stent have been extensively evaluated and have been proven to be significant novel tools for the treatment of coronary artery disease. Several sirolimus derivatives have already emerged, receiving CE Mark approval. However, in the future, it is likely that drugs presently under investigation will address additional mechanisms associated with neointimal formation, either as single agents or in combination with antiproliferative compounds. Concurrently, alterations on stent platform design (helicoidal, open closed cell), coatings (biodegradable, bioabsorbable, nanoporous) and polymers are being explored. PMID- 17076601 TI - Emollient foam in topical drug delivery. AB - Foams offer an innovative and more convenient means of topical drug delivery. The successful introduction of hydroalcoholic foams paved the way for the development of a new generation of foam products that provide skin barrier build-up and hydration. Such foams, designated as emollient foams consist of oil-in-water or water-in-oil emulsions with necessary excipients, such as non-ionic surfactants, gelling agents and foam adjuvants. Emollient foams can carry a broad variety of topical drugs, including water-soluble, oil-soluble and suspended active agents. This paper reviews emollient foam compositions and their physicochemical properties. It further accounts for the usability and functional advantages of emollient foam as a vehicle of topical drugs, including: i) improved usability, which affects treatment, compliance and, consequently, improves therapeutic results; ii) safety; iii) controllable drug delivery; iv) skin barrier build-up and hydration; and v) enhanced clinical efficacy. PMID- 17076602 TI - Progress and potential for regenerative medicine. AB - Regenerative medicine focuses on new therapies to replace or restore lost, damaged, or aging cells in the human body to restore function. This goal is being realized by collaborative efforts in nonmammalian and human development, stem cell biology, genetics, materials science, bioengineering, and tissue engineering. At present, understanding existing reparative processes in humans and exploring the latent ability to regenerate tissue remains the focus in this field. This review covers recent work in limb regeneration, fetal wound healing, stem cell biology, somatic nuclear transfer, and tissue engineering as a foundation for developing new clinical therapies to augment and stimulate human regeneration. PMID- 17076603 TI - Expression of matrix metalloproteinases (MMP-8 and -9) in chronic periodontitis patients with and without diabetes mellitus. AB - BACKGROUND: Matrix metalloproteinases (MMPs) are involved in a number of physiological events, and they are the major players in collagen breakdown during periodontal tissue destruction. Diabetes mellitus (DM) has been associated with altered collagen metabolism and increases the response of the periodontal tissue to pathogenic microorganisms, thereby increasing the severity of periodontal disease. The aim of this study was to assess the expression of MMP-8 and -9 in gingival tissues of diabetic chronic periodontitis (CP), non-diabetic CP, and healthy patients. METHODS: The plaque index (PI), gingival index (GI), probing depth (PD), and clinical attachment level (CAL) were assessed. The levels of MMP expression in gingival tissue extracts were measured by gelatin zymography and Western blotting. The structural changes in tissues were analyzed by histological examination and collagen estimation. RESULTS: Higher levels of PI, GI, CAL, and deeper PD were observed in CP patients with DM than CP patients without DM. The average concentration of MMP-9 was increased three-fold, and the MMP-8 was increased two-fold in CP patients with DM compared to CP patients without DM. A high infiltration of inflammatory cells and less collagen were observed in the histologic analysis of gingival biopsies from diabetic patients. CONCLUSIONS: An increased concentration of MMP-8 and -9 in the gingival tissue of diabetic CP patients suggests that the expression of these MMPs contributes to the failure of the healing process in the diabetic condition. Treatment strategies directed toward the inhibition of these MMPs could lead to an improved healing rate in CP patients. PMID- 17076604 TI - The use of celecoxib and dexamethasone for the prevention and control of postoperative pain after periodontal surgery. AB - BACKGROUND: Conventional non-steroidal anti-inflammatory drugs have been widely used in the control of postoperative pain, but sparse information is available on the efficacy of celecoxib, a selective cyclooxygenase-2 inhibitor, or dexamethasone, a steroidal anti-inflammatory drug, after periodontal surgeries. The purpose of the present study was to compare the use of celecoxib and dexamethasone in the management of pain after mucoperiosteal flap surgery. METHODS: A randomized double-masked cross-over clinical trial was conducted on 20 patients from 27 to 52 years old with generalized moderate to advanced chronic periodontitis. Mucoperiosteal flap surgeries for scaling and root planing were performed under local anesthesia on at least three quadrants, with a 4-week interval between. Each quadrant was randomly assigned to one of the following medication protocols: placebo, 4 mg dexamethasone 1 hour before surgery and 8 hours after the first dose, and 200 mg celecoxib 1 hour before surgery and 12 hours after the first dose. Postoperative pain was accessed during the first 8 hours and on the following 3 days using the visual analog scale (VAS), the 101 point numerical rate scale (NRS-101), and the four-point verbal rating scale (VRS 4). RESULTS: Pain perception was statistically significantly lower in the celecoxib group than in the placebo group during the first 4 hours using VAS (P = 0.01) and at 1, 2, 3, 4, 6, and 7 hours using NRS-101 (P = 0.03). The level of pain was lower in the dexamethasone group than in the placebo only at the 3-hour period (P = 0.001). Statistically significant differences could be found among the groups at 1 hour (P = 0.015), 3 hours (P = 0.004), 4 hours (P = 0.02), and 7 hours (P = 0.05) using VRS-4. There was no statistically significant difference between the celecoxib and dexamethasone groups. CONCLUSION: The findings of this study suggest that the preemptive and postoperative use of celecoxib or dexamethasone were effective in the management of postoperative pain following open-flap debridement. PMID- 17076605 TI - Evaluation of patient perceptions after frenectomy operations: a comparison of carbon dioxide laser and scalpel techniques. AB - BACKGROUND: A frenum that encroaches on the margin of the gingiva may interfere with plaque removal and cause tension. Frenectomy is the complete removal of the frenum that can be made by scalpels or with soft tissue lasers. The aim of this article was to compare the degree of postoperative pain, such as discomfort and functional complications (eating and speech), experienced by patients after two frenectomy operation techniques. METHODS: Forty patients requiring frenectomy were randomly assigned to have treatment either with a conventional technique or with a carbon dioxide (CO2) laser. The postoperative pain and functional complication ratings of each patient were recorded using a visual analog scale on days 1 and 7. RESULTS: The results indicated patients treated with the CO2 laser had less postoperative pain and fewer functional complications (speaking and chewing) (P <0.0001 each) and required fewer analgesics (P <0.001) compared to patients treated with the conventional technique. CONCLUSIONS: This clinical study indicates that CO2 laser treatment used for frenectomy operations provides better patient perception in terms of postoperative pain and function than that obtained by the scalpel technique. Considering the above advantages, when used correctly, the CO2 laser offers a safe, effective, acceptable, and impressive alternative for frenectomy operations. PMID- 17076606 TI - Influence of an erbium, chromium-doped yttrium, scandium, gallium, and garnet (Er,Cr:YSGG) laser on the reestablishment of the biocompatibility of contaminated titanium implant surfaces. AB - BACKGROUND: The aim of the present study was to evaluate the influence of an erbium, chromium-doped yttrium, scandium, gallium, and garnet (Er,Cr:YSGG laser [ERCL]) on 1) the surface structure and biocompatibility of titanium implants and 2) the removal of plaque biofilms and reestablishment of the biocompatibility of contaminated titanium surfaces. METHODS: Intraoral splints were used to collect an in vivo supragingival biofilm on sand-blasted and acid-etched titanium disks for 24 hours. ERCL was used at an energy output of 0.5, 1.0, 1.5, 2.0, and 2.5 W for the irradiation of 1) non-contaminated (20 and 25 Hz) and 2) plaque contaminated (25 Hz) titanium disks. Unworn and untreated non-irradiated, sterile titanium disks served as untreated controls (UC). Specimens were incubated with SaOs-2 osteoblasts for 6 days. Treatment time, residual plaque biofilm (RPB) areas (%), mitochondrial cell activity (MA) (counts per second), and cell morphology/surface changes (scanning electron microscopy [SEM]) were assessed. RESULTS: 1) ERCL using either 0.5, 1.0, 1.5, 2.0, or 2.5 W at both 20 and 25 Hz resulted in comparable mean MA values as measured in the UC group. A monolayer of flattened SaOs-2 cells showing complete cytoplasmatic extensions and lamellopodia was observed in both ERCL and UC groups. 2) Mean RPB areas decreased significantly with increasing energy settings (53.8 +/- 2.2 at 0.5 W to 9.8 +/- 6.2 at 2.5 W). However, mean MA values were significantly higher in the UC group. CONCLUSION: Within the limits of the present study, it was concluded that even though ERCL exhibited a high efficiency to remove plaque biofilms in an energy dependent manner, it failed to reestablish the biocompatibility of contaminated titanium surfaces. PMID- 17076607 TI - In vitro sealing ability of two materials at five different implant-abutment surfaces. AB - BACKGROUND: The aim of the present study was to test the sealing ability of two materials at five different implant-abutment surfaces. METHODS: In the first phase, 2 mul brain-heart infusion (BHI) broth was deposited into the implant wells and glass culture tubes. A varnish or silicon sealant was applied at the cervical implant portion of experimental groups. The control group remained unexposed. The abutments were torque-tightened to 20 Ncm with a manual torque driver. Implants were immersed in 4 ml BHI broth at 37 degrees C for 2 hours to exclude contamination. In the second phase, 100 mul Enterococcus faecalis American Type Culture Collection (ATCC) strain 29212 was deposited into the glass culture tubes. After periods of 7, 14, 21, 35, 49, and 63 days, the sealing capacity was checked. Abutments were removed, and a sterile paper cone collected material inside implant bodies. This material was transferred to new tubes with BHI to verify the presence of cloudy broths within 24 to 48 hours. RESULTS: There were no statistically significant differences between the two materials for each time period (Fisher exact test; P >0.05). Group E showed the least level of sealing ability (six implants contaminated), whereas group T showed the highest level (only two implants). CONCLUSIONS: 1) Materials tested were not able to prevent contamination over 63 days. 2) Bacterial contamination was verified after 14 and 35 days in the control and experimental groups, respectively. 3) Although materials tested had demonstrated similar sealing capacities, dental implants showed bacterial contamination regardless of their external or internal hexagonal configurations. PMID- 17076608 TI - Tuned-aperture computed tomography for detection of induced mid-buccal/lingual alveolar bone defects. AB - BACKGROUND: The detection of bone loss in mid-buccal and lingual crests is impossible using conventional radiographs because of the superimposition of overlying anatomy and lack of three-dimensional information. The purpose of this study was to compare the diagnostic efficacy of tuned-aperture computed tomography (TACT) and conventional two-dimensional direct digital radiography (DDR) in an in vitro environment. METHODS: A total of 45 mandibular molars had 0.8-mm lesions on mid-buccal/lingual crestal areas. Half of the sites received defects, whereas the other half served as controls. Nine DDR images were used to generate TACT slices that were further subjected to iterative restoration (TACT IR). Eight observers used a confidence rating scale to record diagnoses. Receiver operating characteristic (ROC) analysis was done, and areas under the curves were computed (A(z)) as indicators of diagnostic accuracy. Analysis of variance (ANOVA) was used to test for effects of observer, imaging modality, and location on the detection of lesions. RESULTS: TACT-IR performed significantly better than DDR. There was a significant difference in the accuracy of diagnosis based on observers (P <0.001). CONCLUSION: TACT-IR appears to be the imaging modality of choice for the detection of small osseous changes on crestal bone in mid buccal/lingual sites. PMID- 17076609 TI - The effects of interimplant distances on papilla formation and crestal resorption in implants with a morse cone connection and a platform switch: a histomorphometric study in dogs. AB - BACKGROUND: Implant esthetics have been the focus of attention for the last few years, and one of the most important points is the effect that interimplant distances can have on papilla formation and bone loss. The aim of this study was to evaluate the effect that distances of 1, 2, and 3 mm between implants after prosthetic restoration will have on crestal bone resorption (from the top of the implant to the bone crest [TI-BC]) and bone resorption (from the top of the implant to the first bone-to-implant contact TI-BIC) in two-stage implants used in a submerged and non-submerged protocol. METHODS: The mandibular bilateral premolars of seven dogs were extracted, and after 12 weeks, each dog received eight implants. The implants were placed so that three interimplant contact points were created, with 1-mm (group 1), 2-mm (group 2), and 3-mm (group 3) distances constructed on each side. The sides and the position of the groups were randomly selected. After 12 weeks, the implants received metallic prostheses with 5 mm between the contact point and the bone crest. After 8 weeks more, the animals were sacrificed. RESULTS: The TI-BC was 0.20 and 0.18 mm for group 1, 0.15 and 0.14 mm for group 2, and 0.15 and 0.15 mm for group 3 for non-submerged and submerged implants, respectively. At the proximal region, the TI-BC was 0.16 mm for non-submerged and 0.16 mm for submerged implants. The TI-BIC was 0.32 and 0.30 mm for group 1, 0.19 and 0.21 mm for group 2, and 0.30 and 0.24 mm for group 3 for non-submerged and submerged implants, respectively. At the proximal region, the TI-BIC was 0.26 mm for non-submerged and 0.25 mm for submerged implants. There was no statistical difference for any of the parameters (analysis of variance [ANOVA]). CONCLUSION: Distances of 1, 2, and 3 mm between implants do not result in statistically significant differences on TI-BC and TI-BIC around submerged or non-submerged implants with a Morse cone connection and a platform switch. PMID- 17076610 TI - Functional polymorphisms in the matrix metalloproteinase-9 gene in relation to severity of chronic periodontitis. AB - BACKGROUND: Members of the matrix metalloproteinase (MMP) family are implicated in the chronic remodeling in periodontal diseases. Therefore, we performed a case control study to investigate a plausible association between susceptibility to chronic periodontitis (CP) and the polymorphisms in the MMP-9 (gelatinase B) gene. METHODS: Using polymerase chain reaction with subsequent restriction analyses, MMP-9 -1562C/T and R+279Q variants were determined in 304 subjects (169 patients with CP, 76 with a mild to moderate form and 93 with severe generalized CP, and 135 age- and gender-matched unrelated control subjects). RESULTS: The distribution of the MMP-9 -1562C/T and R+279Q genotypes and alleles did not significantly differ between cases and controls. However, the frequency of the T variant at position -1562 was marginally higher in patients with severe disease compared to those with mild to moderate forms (P <0.05; P(corr) >0.05). In further analysis, the -1562T allele was associated with a severe form of CP in men (odds ratio: 3.87; 95% confidence interval: 1.40 to 10.65; P <0.01 and P(corr) <0.05 for allele; P <0.05 and P(corr) >0.05 for genotype) but not in women. CONCLUSION: These findings suggest that genetic polymorphisms of the MMP-9 gene are not associated with the susceptibility to CP in the Czech population; however, the promoter variant may influence the severity of the disease in men. PMID- 17076611 TI - Preprosthetic periodontal surgery in the proximal area with modification of the col area: results following the reestablishment of the contact point. AB - BACKGROUND: Anatomic characteristics of interproximal areas are dependent on the anatomy, position, and proximal contact of adjacent teeth. The objective of this study was to investigate the influence of the reestablishment of the interproximal contact following the restorative alveolar interface (RAI) procedure on the interproximal gingival COL and formation of the interdental gingival papilla. METHODS: Six mongrel dogs received bilateral apically positioned flaps, crown lengthening, and the RAI procedure on the maxillary fourth bicuspid and first molar. After 2 weeks, in a randomized manner, one side was prepared to receive metallic crowns and the opposite side remained as the control. The crowns were cemented at the 4-week postoperative period, and the dogs were sacrificed after another 4 weeks, totaling a period of 4 weeks with the full crowns in position and a total of 8 postoperative weeks. Histologic specimens were stained with hematoxylin and eosin and Mallory dyes. Sections 6 micro m thick were obtained in a bucco-lingual plane allowing ample visualization of the interproximal area. RESULTS: Clinical measurements revealed that, in the restored sides, four animals had complete fill of the interdental spaces with gingival papilla, whereas the other two dogs had a distance from the contact point to the tip of papilla varying from 0.02 to 0.021 mm. In the control group, papillae were totally reepithelialized with keratinized epithelium and a convex form. The epithelium completely covered the connective tissue and showed both epithelial projections and surface desquamation. On the test group, despite the presence of the prosthesis, the COL morphology modified by preprosthetic surgery was not altered, presenting a convex papilla with a triangular form and with a keratinized epithelium. Additional histologic characteristics were the same as found in the control group. CONCLUSION: Based on the results of this study, the reestablishment of the contact point does not revert what was obtained with the RAI procedure; the interproximal tissues remain convex and keratinized. PMID- 17076612 TI - Matrix metalloproteinase-13 is highly expressed in destructive periodontal disease activity. AB - BACKGROUND: Matrix metalloproteinases (MMPs) participate in extracellular matrix degradation in physiological and pathological conditions. The available evidence suggests that MMP-13 plays a significant role in both the initiation and progress of bone resorption. The aim of our study was to identify the presence of MMP-13 in adult patients with untreated chronic periodontitis. We also determined the activity of MMP-13 present in lesions undergoing episodic attachment loss in gingival crevicular fluid (GCF) samples. METHODS: After monitoring at 2 and 4 months, 21 patients showed destructive periodontitis (periodontally affected sites presenting at least two sites with > or =2 mm clinical attachment loss), and GCF samples were collected both from active and inactive sites (21 GCF samples, each). GCF was collected during a 30-second interval using a paper strip, and an immunofluorescence assay was performed to determine the basal activity of MMP-13 and the relationship between 4-aminophenylmercuric acetate (APMA)-activated total MMP-13 and basal MMP-13 activity. Gingival tissues from five patients were fixed in formalin and MMP-13 expression was demonstrated using immunohistochemistry and in situ hybridization. MMP-13 molecular forms were examined by Western immunoblotting with monoclonal antibodies. RESULTS: MMP-13 was found in 100% of GCF samples from patients with chronic periodontitis. Active sites, associated with tissue destruction, had significantly higher proportions of active MMP-13 and MMP-13 activity levels than their inactive counterparts (1.49 versus 1.17 ng fluorescent product, respectively; P <0.05). Western blot, immunohistochemical staining, and in situ hybridization confirmed the presence of MMP-13 in periodontal disease, with observable differences between periodontitis and healthy subjects. MMP-13 immunoreactivities were seen mainly as 55 and 48 kDa, corresponding to partially and fully activated forms, respectively, and a smaller proportion of 60-kDa proenzyme form. CONCLUSION: MMP-13 activity in GCF samples was significantly increased in active sites from progressive periodontal disease, supporting its role in the alveolar bone loss developed in this disease. PMID- 17076613 TI - Elevated serum interleukin-6 (IL-6) in solid-organ transplant recipients is positively associated with tissue destruction and IL-6 gene expression in the periodontium. AB - BACKGROUND: The number of transplanted solid organs and life expectancy after transplantation are steadily rising worldwide. Inflammation is widely recognized as playing a pivotal role in transplant rejection, and several studies have shown that serum interleukin-6 (IL-6) levels can identify individuals who are at greater risk for rejection. Given the known association between IL-6 and chronic periodontitis, the aim of our study was to assess the periodontal status of solid organ transplant subjects compared to systemically healthy controls, to quantify the IL-6 levels in the serum and periodontal tissues, and to explore their association. METHODS: Forty-seven heart and kidney transplant and 18 systemically healthy age-matched individuals were recruited. Subjects received a complete periodontal examination, and blood and periodontal tissue samples were collected for quantification of IL-6 protein and mRNA levels, respectively. RESULTS: Transplant subjects had significantly higher serum IL-6 levels and slightly but statistically significantly increased mean probing depths than healthy controls. Multivariable linear regression analysis adjusting for gender, diabetes, smoking, and immunosuppressant dose showed that the mean probing depth, number of missing teeth, and mean percentage of sites with > or =4 mm attachment loss were independent predictors for elevated serum IL-6 levels. Transplant subjects with chronic periodontitis had higher mean serum IL-6 levels than those without chronic periodontitis, and there was a positive correlation between periodontal IL-6 gene expression levels and serum IL-6 protein levels. CONCLUSIONS: Periodontal tissue destruction and local IL-6 synthesis are associated with elevated serum IL-6 levels in transplant recipients. This may have serious implications in solid-organ transplant deterioration and chronic rejection. PMID- 17076614 TI - Treatment of intrabony defects with an enamel matrix protein derivative or bioabsorbable membrane: an 8-year follow-up split-mouth study. AB - BACKGROUND: Treatments with either an enamel matrix protein derivative (EMD) or guided tissue regeneration (GTR) have been shown to promote periodontal regeneration. However, until recently, only limited data have been available on the long-term clinical results following these regenerative techniques. Therefore, the aim of this study was to present the 8-year results of a prospective, controlled, split-mouth clinical study evaluating the treatment of intrabony defects with EMD or GTR. METHODS: Ten patients, each of whom displayed one pair of intrabony defects located contralaterally in the same jaw, were randomly treated with EMD or with GTR by means of bioabsorbable membranes. The following clinical parameters were evaluated at baseline and at 1 and 8 years after treatment: plaque index (PI), gingival index (GI), bleeding on probing (BOP), probing depth (PD), gingival recession (GR), and clinical attachment level (CAL). The primary outcome variable was CAL. No statistically significant differences between the groups were found at baseline. RESULTS: The sites treated with EMD demonstrated a mean CAL change from 9.5 +/- 1.2 mm to 6.3 +/- 1.3 mm (P <0.001) and 6.7 +/- 1.6 mm (P <0.001) at 1 and 8 years, respectively. No statistically significant differences were found between the 1- and 8-year results. Sites treated with GTR showed a mean CAL change from 9.7 +/- 1.3 mm to 6.7 +/- 0.9 mm (P <0.001) at 1 year and 6.8 +/- 1.2 mm (P <0.001) at 8 years. The CAL change between 1 and 8 years did not present statistically significant differences. Between the treatment groups, no statistically significant differences in any of the investigated parameters were observed at 1 and at 8 years. However, the study does not have the statistical power to rule out the possibility of a difference between the two groups. CONCLUSIONS: Within their limits, the present results indicate the following: 1) the clinical improvements obtained following treatment with EMD or GTR can be maintained over a period of 8 years; and 2) further studies of much higher power need to be performed to support equivalence. PMID- 17076615 TI - Hypogalactosylation of salivary and gingival fluid immunoglobulin G in patients with advanced periodontitis. AB - BACKGROUND: Altered glycosylation of immunoglobulin G (IgG) has been found to affect certain immunological activities of IgG and to correlate with increased inflammation in various disease states. This work deals with the changes in distribution and galactosylation of IgG subclasses present in saliva and gingival crevicular fluid (GCF) of patients with initial and advanced periodontitis and of normal controls. METHODS: IgG subclasses were quantified by dot-blot assay, and the degrees of expression of galactose in the total IgG and its individual subclasses were estimated by lectin immunoblot assay after sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) separation of IgG and by capture enzyme-linked immunosorbent assay (ELISA), using biotinylated Ricinus communis (RCA-I) and Bandeiraea simplicifolia (BS-II) lectins. RESULTS: The distribution of IgG subclasses in both fluids was found to differ in periodontal patients compared to normal controls. In the periodontitis saliva and GCF, the IgG2 subclass dominated quantitatively, regardless of periodontal status. However, galactose was found to be expressed in IgG heavy chains in normal controls and patients with initial periodontitis but not, or at barely detectable levels, in advanced periodontitis. CONCLUSION: The results suggest that the shift toward hypogalactosylated glycoforms may occur during the process of inflammation of the gingiva. PMID- 17076616 TI - New evidence of premature oxidative DNA damage: mitochondrial DNA deletion in gingival tissue of patients with periodontitis. AB - BACKGROUND: Overproduction of reactive oxygen species (ROS) causes increased oxidative stress in gingival tissue. It has been generally accepted that increased oxidative stress might contribute to additional damage of lipids, proteins, and DNA molecules. The mitochondrial DNA (mtDNA) mutation is a superb biomarker of oxidative damage. The aim of the present study was to investigate the mtDNA deletions in the gingival tissue of patients with periodontitis and to explain the correlations between mtDNA deletion in gingival tissue and clinical parameters of periodontitis and age. METHODS: Gingival tissue and blood samples were collected from 30 patients with chronic periodontitis (CP group) and 30 healthy control subjects (H group). To determine the clinical condition of each subject, the plaque index, gingival index, clinical attachment level, and probing depth were measured. Using the polymerase chain reaction (PCR) method, we examined the 7.4- and 5-kbp mtDNA deletions in tissue and blood samples. Three different pairs of PCR primers were used in this study. RESULTS: In this study, we did not detect any deletions in blood DNA samples in either the CP or H group. Also, the 7.4-kbp mtDNA deletion was not detected in gingival tissues of subjects. However, the 5-kbp mtDNA deletion was detected in 24 of the 30 subjects (80%) in the CP group and was not detected in the H group (0%). Significant correlations were found between the occurrence of the 5-kbp mtDNA deletion and all clinical parameters (P <0.01). A similar correlation was found between the occurrence of the 5-kbp mtDNA deletion and age (P <0.05). CONCLUSIONS: The overproduction of ROS by activated polymorphonuclear leukocytes in chronic inflammation may lead to premature oxidative damage of the mtDNA. In this study, the occurrence of the 5-kbp mtDNA deletion in 24 periodontitis subjects may be evidence of premature oxidative DNA damage. PMID- 17076617 TI - Modified coronally advanced flap associated with a subepithelial connective tissue graft for the treatment of adjacent multiple gingival recessions. AB - BACKGROUND: The clinical choice of the appropriate surgical technique aiming at root coverage relies, among other factors, on the number of adjacent gingival recessions. This study aimed to clinically evaluate the effectiveness and the predictability of root coverage at adjacent multiple gingival recessions using a modified coronally positioned flap associated with the subepithelial connective tissue graft. METHODS: Ten non-smoking, healthy subjects (five men and five women; mean age, 28.7 years) presenting 29 Class I or II adjacent multiple gingival recessions were enrolled. Each patient was treated using a modified coronally advanced flap associated with the subepithelial connective tissue graft. Probing depth (PD), clinical attachment level (CAL), recession depth (RD), and width of keratinized tissue (KT) were measured at baseline and 6 months later. The Student t test was used to compare treatment outcomes through time. RESULTS: The results revealed significant CAL gain (mean gain +/- SD, 1.97 +/- 0.94 mm; P <0.0001), RD decrease (2.03 +/- 0.78 mm; P <0.0001), and KT increase (1.31 +/- 1.23 mm, P <0.0001). The average root coverage was 96.7%, and complete root coverage was found at 93.1% of the defects. Nine of the 10 patients (90% of the patients) experienced complete root coverage. CONCLUSIONS: The modified coronally advanced flap associated with the subepithelial connective tissue graft was effective and predictable to produce root coverage at multiple adjacent gingival recessions associated with gain in the CAL and in the width of KT. PMID- 17076618 TI - Decreased lipid peroxidation following periodontal therapy in type 2 diabetic patients. AB - BACKGROUND: Although diabetes mellitus and periodontal disease promote atherosclerosis, the relation of oxidative stress with these diseases remains unclear. To investigate the influence of periodontal disease on oxidative stress, we assessed the effects of initial periodontal therapy on lipid peroxide (LPO), an oxidative stress index, in type 2 diabetic and non-diabetic patients. METHODS: Seventeen subjects with or without type 2 diabetes were enrolled in this intervention study. No patient had a history of cardiovascular or peripheral vascular disease. Five type 2 diabetic and six non-diabetic patients, all with moderate to severe periodontal disease, received and completed the initial periodontal therapy and examination. Before and after the therapy, patients underwent medical examinations and blood determinations, including LPO. RESULTS: Before the therapy, the periodontal probing depth and bleeding on probing (BOP) were similar between groups. LPO, triglyceride, and white blood cell counts were significantly higher in diabetic than non-diabetic patients. Therapy improved the periodontal parameters in both groups and significantly decreased LPO in diabetic patients. Anti-malondialdehyde-modified low-density lipoprotein (MDA-LDL) antibody, a marker of oxidized LDL, significantly decreased with treatment in both groups. Overall, Spearman rank correlation showed no significance between periodontal parameters and LPO or anti-MDA-LDL antibody, but BOP tended to correlate with LPO in diabetic patients (r = 0.585; P = 0.0791). CONCLUSION: Although this is a small and preliminary study, and the changes of LPO and anti MDA-LDL antibody were within the normal range, the initial periodontal therapy significantly decreased LPO, an oxidative stress index, in type 2 diabetic patients with periodontal disease. PMID- 17076619 TI - Immediate restoration of single implants placed immediately after tooth extraction. AB - BACKGROUND: Some of the original surgical protocols for implant surgery have been reassessed to satisfy the patient's continuously increasing expectation for shorter rehabilitation time, improved esthetics, and increased comfort. The aim of this study was to evaluate the 12-month clinical success of 18 single-tooth implants inserted immediately after tooth extraction and restored the same day with non-functional loading. METHODS: A total of 18 patients (12 women and six men), 22 to 60 years old, were enrolled in this study. Eighteen single implants were inserted in fresh extraction sockets and immediately restored with temporary abutments and crowns. All experimental sites showed an absence of fenestrations or dehiscences of the bone walls and a residual gap between implant surfaces and surrounding bone walls < or =2 mm. All temporary prostheses were positioned the same day of surgery and were not in occlusal contact with opposing arches. The comparison between the baseline and 12-month visits was performed with the Student t test for paired data (statistically significant at a level of alpha = 0.05). RESULTS: During the 12-month follow-up period, one fixture was removed 4 weeks after implant placement following an abscess. All remaining implants healed uneventfully with no complications and were assessed as stable and successful at the 12-month checkup. No technical complications such as screw loosening, resin fracture, or pain during chewing were registered during the 12-month period. CONCLUSIONS: Within the limits of the data from this study, it can be suggested that immediate placement and restoration of a single implant can be a valid and successful option of treatment in the case of single compromised teeth. Moreover, this treatment protocol eliminates the need for removable provisional restoration and seems to maintain the preexisting architecture of soft and hard tissues in most cases. Nonetheless, further prospective and long-term studies are required to obtain a better insight into the limitations of this protocol. PMID- 17076620 TI - Periodontitis as a manifestation of chronic benign neutropenia. AB - BACKGROUND: A subcategory of chronic neutropenia is chronic benign neutropenia, which is characterized by a prolonged non-cyclic neutropenia as the sole abnormality, with no underlying disease to which the neutropenia can be attributed. Chronic neutropenia is defined as a low absolute neutrophil count for >6 months. In this presentation, periodontitis seems to be the sole manifestation of a juvenile patient with chronic benign neutropenia. A 7-year-old white male presented with periodontitis of the primary dentition and early tooth loss. His medical and dental history was otherwise unremarkable. Suspecting some systemic illness as the underlying cause, the patient was referred for a medical consultation and a series of blood tests. METHODS: Blood analyses included a complete blood count (CBC), sequential multiple analyzer 24 (SMA 24), glycated hemoglobin levels, and screening for anti-white blood cell antibodies. Blood levels of calcium, vitamin D, dihydroxyvitamin-D, phosphorus, and alkaline phosphatase were also measured. Liver function tests were performed. RESULTS: Following analysis of recent and previous blood test results, a diagnosis of chronic benign neutropenia was assigned. The patient's periodontal condition was treated with scaling and root planing, oral hygiene instruction, and antimicrobial mouthrinses. Three-month recall visits were recommended as a follow up protocol. CONCLUSIONS: This case represents the importance of diagnosing periodontal disease as a possible indicator of underlying systemic disease. When a patient presents with an unusual, generalized form of periodontal disease, screening for systemic disorders is required, as the oral condition may be the first or only manifestation of a systemic abnormality. This case also illustrates the reason for the change in classification of such a condition to periodontitis as a manifestation of systemic disease. This condition was previously classified as prepubertal periodontitis, a disease diagnosis that focused on the patient's age at the onset of the disease rather than the etiology. PMID- 17076621 TI - President's address. PMID- 17076623 TI - Preclinical pharmacology of AL-12182, a new ocular hypotensive 11-oxa prostaglandin analog. AB - PURPOSE: The aim of this study was to determine selected in vivo ocular properties of AL-12182 (5,6-dihydro-4,5-didehydro-11-deoxy-11-oxa-16-(3 chlorophenoxy)-omega-tetranor-PGF(2alpha) isopropyl ester) and the in vitro profile of its free acid, AL-12180. METHODS: Previously documented radioligand binding and functional assays involving human ciliary muscle cells (h-CM), human trabecular meshwork (h-TM) and other cells, and porcine ocular arteries were utilized. For in vivo procedures, we utilized rabbits, cats, and nonhuman primates to measure hyperemia, pupil diameter, and intraocular pressure (IOP), respectively. RESULTS: AL-12180 exhibited the highest affinity for the FP receptor (K(i) = 143 +/- 36 nM) and much lower affinity for DP-, EP(3)-, IP-, and TP-receptors, and for several nonprostanoid receptors, enzymes, neurotransmitter uptake sites, ion channels, and other regulatory sites. AL-12180 activated phospholipase C-mediated phosphoinositide hydrolysis (potency, EC(50) = 13.7-42.7 nM) through the FP-receptor in a variety of cells, such as h-CM, h-TM cells, human embryonic kidney cells expressing the cloned human ciliary body FP-receptor (HEK-FP), mouse 3T3 cells, and rat vascular smooth muscle cells. AL-8810, an FP antagonist, blocked the effects of AL-12180 in h-CM cells (IC(50) = 8.7 microM). AL-12180 also stimulated the mobilization of intracellular Ca(2+) ([Ca(2+)](i)) in h-TM cells (EC(50) = 111 +/- 36 nM), h-CM cells (EC(50) = 11 nM), and in host cells expressing the cloned human ciliary body FP-receptor (EC(50) = 5.9 +/- 3.1 nM). AL-12180 lacked significant agonist activity at DP-, EP(2)-, EP(4)-, IP-, and TP-receptors in cell-based assays. However, AL-12180 contracted porcine central retinal and short posterior ciliary arteries in vitro with micromolar potencies that appeared to involve TP-receptor activation. in vivo, AL-12182 elicited dose-related hyperemia in the rabbit eye, miosis in the cat eye, and ocular hypotension in the nonhuman primate eye. CONCLUSIONS: AL-12180 is a relatively potent and selective FP-receptor agonist whose isopropyl ester prodrug (AL-12182) lowers IOP by as much as 40% following topical ocular dosing in a laser-induced nonhuman primate model of ocular hypertension. PMID- 17076624 TI - Kinin modulation of conventional outflow facility in the bovine eye. AB - The aim of this study was to investigate the effects of bradykinin on conventional outflow facility in relation to kinin effects on matrix metalloproteinase (MMP) secretion. Conventional outflow facility was measured in isolated bovine segments perfused at a constant pressure of 10 mmHg. Experiments were also performed in primary cultures of bovine trabecular meshwork cells to assess bradykinin effects on the secretion of MMP-9 assessed by western blot. Administration of bradykinin (10(-7) M) to perfused anterior segments produced a 50% increase in outflow facility above basal levels. The effect was slow to develop, requiring 100 min for a significant increase in facility and 4 h for the peak response to be observed. Pretreatment of anterior segments with the B(2) kinin receptor antagonist, HOE-140 (10(-6) M), or with the nonselective MMP inhibitor, GM6001 (10(-5) M) blocked the response to bradykinin (10(-7) M). Treatment of cultured trabecular meshwork cells with bradykinin (10(-7) M) for 120 min stimulated secretion of MMP-9 into the extracellular media, and this response was inhibited by HOE-140 (10(-6) M). These results demonstrate that bradykinin activates B(2) kinin receptors to increase conventional outflow in the perfused bovine eye and provide evidence that secretion and activation of MMPs within the conventional pathway may mediate the effect. PMID- 17076625 TI - Vasodilator effects of adrenomedullin on retinal arterioles in streptozotocin induced diabetic rats. AB - PURPOSE: The aim of this study was to examine whether diabetes changes the in vivo effects of adrenomedullin (ADM) on diameter of retinal arteriole and blood pressure in rats. METHODS: Male Wistar rats were treated with streptozotocin (65 mg/kg, intravenously [i.v.]) and experiments were performed 6-8 weeks later. Under artificial ventilation, rats were injected with tetrodotoxin (50 microg/kg, i.v.) to eliminate any nerve activity and prevent movement of the eye. A mixture solution of norepinephrine and epinephrine (1:9) was used to maintain adequate systemic circulation. Diameters of retinal vessels were measured from the fundus images and were captured by a digital camera that was equipped with a special objective lens. RESULTS: ADM dose-dependently increased diameters of retinal arterioles and decreased blood pressure in streptozotocin-induced diabetic rats and the age-matched controls. The depressor responses, but not vasodilator responses of retinal arterioles, to ADM were reduced in diabetic rats. CONCLUSIONS: These results suggest that mechanism(s) of ADM-induced vasodilation of retinal arterioles is preserved in diabetes, even when depressor effects of ADM are impaired. ADM may play a role as a regulatory mechanism of retinal circulation in nondiabetic and diabetic conditions. PMID- 17076626 TI - Development of miotic cross-tolerance between pyridostigmine and sarin vapor. AB - The organophosphorous nerve agent sarin (GB) and the carbamate pyridostigmine bromide (PB) both inhibit acetylcholinesterase (AChE), leading to overstimulation of muscarinic receptors. Both GB and PB produce miosis through stimulation of ocular muscarinic receptors. This study investigated 2 hypotheses: (1) that the miotic response to PB would decrease following repeated injections; and (2) that repeated administration of PB would result in tolerance to the miotic effect of GB vapor. Rats were injected intramuscularly with saline, 0.04 mg/kg, 0.5 mg/kg, or 1.4 mg/kg of PB twice daily for 8 consecutive days. After day 3, animals injected with 1.4 mg/kg PB developed miotic tolerance. Twenty-four (24) h following the final PB injection, the rats were exposed to GB vapor (4.0 mg/m(3)). A similar magnitude of miosis was observed in all groups after GB exposure. However, the rate of recovery of pupil size in animals pretreated with 0.5 and 1.4 mg/kg PB was significantly increased. Twenty (20) h following exposure to GB vapor, the pupils of animals pretreated with 1.4 mg/kg PB had recovered to 77% +/- 4% of their pre-exposure baseline, whereas the saline injected controls had recovered to only 52% +/- 2% of their pre-exposure baseline. The increased rate of recovery does not appear to be a result of protection of pupillary muscarinic receptors by the higher doses of PB, as there was no longer PB present in the animal at the time of GB exposure. These results demonstrate the development of tolerance to the miotic effect of PB following repeated exposures, and also suggest that cross-tolerance between PB and GB occurs. However, because the magnitude of the response was not reduced, the PB pretreatment and its associated miotic cross-tolerance does not appear to diminish the effectiveness of miosis as a biomarker of acute exposure to nerve agent vapor. PMID- 17076627 TI - Improvement of the ocular bioavailability of carteolol by ion pair. AB - Ocular bioavailability after instillation of carteolol was investigated by ion pair formation, taking into consideration a balance between lipophilicity and water solubility. The octanol/ water partition coefficient (PC(O/W)) and the aqueous humor concentration in rabbits after instillation of carteolol containing fatty acids having not more than 6 carbons were measured. The longer carbon chain fatty acid showed the higher PC(O/W) of carteolol. The aqueous humor concentration of carteolol increased with carbon chain length of fatty acid and was clearly correlated with logPC(O/W). The increment of counter ion also increased both the logPC(O/W) and aqueous humor concentration of carteolol. The findings suggested that the transcorneal absorption of carteolol would be designed by coordinating with quality and quantity of counter ions. The area under concentration (AUC) in aqueous humor applied by ion pair formulation containing 2% carteolol with sorbate was 2.6 times higher than that by 2% carteolol ophthalmic solution (control), whereas the AUC applied by 4% carteolol ophthalmic solution was 1.4 times higher. The plasma level after instillation of ion pair formulation was almost the same as that of 2% ophthalmic solution. The ratio of AUC (aqueous humor/ plasma) of ion pair formulation was markedly higher, as compared with those of 2% and 4% ophthalmic solution. These results showed that the ion pair formation with sorbate improved the ocular bioavailability of carteolol without enhancing systemic absorption. PMID- 17076628 TI - Ocular powder: dry topical formulations of timolol are well tolerated in rabbits. AB - PURPOSE: Although eye drops are the most common form of ocular drugs, they have several limitations. Drug absorption into the eye is, in general, less than 5%, addition of preservatives is often necessary, and many drugs cannot be formulated as eye drops. Formulating ocular drugs as powder may solve these problems. The aim of this study was to investigate ocular irritation in rabbits following powder administration. METHODS: Timolol maleate (TM) powder was administered to pigmented lop rabbits. Both pure TM powder and freeze-dried with PVP-polymer (2.4% of mass) were tested in 1.0- and 0.1-mg doses. Additionally, 4 rabbits received 0.1 mg of the pure powder 3 times a day for 8 d. Redness of the bulbar conjunctiva and the amount of discharge was rated from photographs (0-3 points, randomized and masked evaluation). The 8-d experiment additionally included examination with a slit lamp and examination of hematoxylin-eosin stained sections of eyes with light microscopy. RESULTS: No serious or irreversible signs of irritation were noted. Doses of 1.0 mg were more irritating than 0.1-mg doses. There was no detectable difference in irritation between pure or freeze-dried powder. Slit-lamp examination, surface photographs and histology showed a negligible difference between drug and control eyes following the 8-d experiment. CONCLUSIONS: The results suggest that 0.1 mg of timolol powder does not irritate the eye and that testing topical timolol powder in humans is feasible. PMID- 17076629 TI - Pharmacokinetic analysis of platelet-activating factor in the tears of guinea pigs with allergic conjunctivitis. AB - PURPOSE: The aim of this study was to examine the levels of platelet-activating factor (PAF) and lyso-PAF in tears from experimental animals developing allergic conjunctivitis (AC). METHODS: AC was induced in guinea pigs by application of ovalbumin in eye drops. Tear samples were collected from 5 actively sensitized animals and from 5 unsensitized control animals before the challenge, and 1, 2, 4, and 6 h postchallenge. C18:0-PAF, C18:0-lyso-PAF, C16:0-PAF, and C16:0-lyso PAF levels in the tear samples were determined using liquid chromatography-tandem mass spectrometry. RESULTS: The concentrations of C16:0-PAF, C16:0-lyso-PAF, and C18:0-lyso-PAF were measurable in both unsensitized and sensitized groups, whereas C18:0-PAF was undetectable in tear samples from either group. The levels of C16:0-PAF, C16:0-lyso-PAF, and C18:0-lyso-PAF in sensitized animals increased throughout the time course of the experiment, whereas there was no corresponding increase in the levels of these molecules in the unsensitized group. There were strong correlations between the concentrations of C16:0-PAF and C16:0-lyso-PAF, both in the sensitized and in the unsensitized group, and the concentrations of C16:0-lyso-PAF and C18:0-lyso-PAF within each group. CONCLUSIONS: The data from this study demonstrated that the levels of PAF and lyso-PAF increase in tears in a guinea pig model of AC development and implicate a role for PAF for the development of AC. PMID- 17076630 TI - A comparative analysis of the effects of the fixed combination of timolol and dorzolamide versus latanoprost plus timolol on ocular hemodynamics and visual function in patients with primary open-angle glaucoma. AB - AIMS: The aim of this study was to assess the effects of fixed combination of timolol and dorzolamide and latanoprost plus timolol on retinal, choroidal, and retrobulbar hemodynamics and visual function in primary open-angle glaucoma (OAG) subjects. METHODS: Sixteen (16) OAG patients (age, 63.5 +/- 10.8 years; 9 male) were evaluated in a randomized, crossover, double-blind study design after 4 weeks of treatment of latanoprost with timolol and fixed combination of timolol and dorzolamide. After randomization, 9 right eyes and 7 left eyes were included in the hemodynamic portion of the study. Measurements included: adverse events check, visual acuity, contrast sensitivity, blood pressure, heart rate, intraocular pressure (IOP), and fundus examination. Ocular blood flow was assessed using confocal scanning laser Doppler flowmetry, color Doppler imaging, and scanning laser ophthalmoscopy. RESULTS: Both therapies were effective at lowering IOP, whereas there was no statistically significant difference between latanoprost plus timolol and the fixed combination of timolol and dorzolamide (13.9% and 12.2% reduction, respectively; P = 0.5533). Fixed combination of timolol and dorzolamide significantly increased central retinal artery end diastolic blood flow velocity (P = 0.0168) and lowered resistance to flow (P = 0.0279). Temporal posterior ciliary artery peak systolic and end diastolic velocities were significantly increased with the fixed combination of timolol and dorzolamide (P = 0.0125 and 0.0238, respectively). Latanoprost plus timolol had no significant effects on ocular blood flow during 4 weeks of treatment. There were no statistically significant differences in adverse events, blood pressure, heart rate, visual acuity, contrast sensitivity scanning laser ophthalmoscopy, or Heidelberg Retinal Flowmeter for any treatment period. CONCLUSIONS: Fixed combination of timolol and dorzolamide therapy might increase blood flow in OAG patients while attaining a similar IOP reduction compared to latanoprost plus timolol. Visual function, however, was not different in this short-term comparison between the two treatments. PMID- 17076631 TI - The effect of recombinant human hyaluronidase on dexamethasone penetration into the posterior segment of the eye after sub-Tenon's injection. AB - PURPOSE: The aim of this study was to investigate the extent if recombinant human hyaluronidase (rhuPH20) can enhance trans-scleral penetration of sub-Tenon's dexamethasone (DM) into the posterior segment of the eye. METHODS: rhuPH20 was purified from conditioned media through a series of ion exchange, hydrophobic interaction, aminophenylboronate, and hydroxyapatite chromatography to greater than 90% purity based upon specific activity. Only the right eye of each rabbit was injected. The first group (n = 16) received an injection of DM and rhuPH20, whereas the second group (n = 16) received DM only. The eyes were enucleated 1, 2, 3, and 6 h after the injection, and the choroid, retina, vitreous, aqueous, and serum were harvested. DM concentration was assessed by mass spectrometry. Histology (n = 2) and immunohistochemistry (n = 2) was performed to detect toxicity and the presence of the rHuPH20, respectively. RESULTS: We observed no histopathologic damage to ocular tissues after sub-Tenon's injection. This enzyme significantly increased DM level in the choroid and the retina 3 h after administration. The rise in levels was transient returning to normal levels by 6 h. CONCLUSIONS: Sub-Tenon's coinjection of rHuPH20 with DM resulted in a general increase in DM levels in ocular tissues and the serum, with significant increase in the choroid and the retina, 3 h after administration. PMID- 17076632 TI - Visual acuity change after intravitreal triamcinolone in various types of exudative age-related macular degeneration. AB - OBJECTIVE: The aim of this study was to compare the visual acuity change after an intravitreal high-dose injection of triamcinolone acetonide (TA) in various types of exudative age-related macular degeneration (AMD). PARTICIPANTS: The interventional comparative case series study included 142 patients (146 eyes) with progressive exudative AMD differentiated into the occult type (n = 78; 53.4%), minimal classic type (n = 45; 30.8%), predominantly classic type (n = 17; 11.6%), and the purely classic type (n = 6; 4.1%). Mean follow-up was 9.7 +/- 7.0 months (3-35.7 months). METHODS: Single intravitreal injection of approximately 20 mg of TA. OUTCOME MEASURES: Visual acuity, intraocular pressure (IOP). RESULTS: Gain in visual acuity measured at 1 month (P = 0.20), 2 months (P = 0.43), and at 3 months (P = 0.38) after the intravitreal injection of triamcinolone and maximal gain in visual acuity during the whole follow-up (P = 0.81) did not vary significantly between the 4 study groups. Correspondingly, the size of a retinal pigment epithelium detachment was not significantly associated with the change in visual acuity at 1 month (P = 0.62), 2 months (P = 0.24), 3 months (P = 0.96), or the maximal gain in visual acuity during follow-up (P = 0.93). The amount of rise in IOP, compared with the baseline value (6.5 +/- 7.4 mmHg), was statistically not associated with the type of subfoveal membrane (P = 0.20; 95% confidence interval: -0.52, 2.45). CONCLUSIONS: The change in visual acuity and the rise in IOP in patients with exudative AMD receiving an intravitreal triamcinolone monotherapy is statistically independent of the type of subfoveal membrane, including the size of a retinal epithelium detachment. PMID- 17076633 TI - Corneal safety of topically applied naltrexone. AB - PURPOSE: Naltrexone (NTX), an opioid antagonist, accelerates wound healing of corneal epithelium in normal and diabetic animals. This study examined the safety of NTX by topical application in the cornea. METHODS: NTX in concentrations of 10(-3), 10(-4), 10(-5), 10(-6), or 10(-7) M was administered topically 4 times/daily for 7 d to the eye of Type 1 diabetic rats (glucose levels >400 mg/dL) (DB), DB animals receiving insulin to maintain normoglycemia (DB-IN), and normal (nondiabetic) (Normal) subjects beginning 8 weeks after onset of diabetes. RESULTS: No differences in intraocular pressures, corneal thickness, endothelial cell number, or epithelial apoptosis, necrosis, or organization were observed between DB, DB-IN, and Normal groups with and without treatment with NTX. The DB group had a twofold decrease in corneal sensitivity from the Normal and DB-IN groups prior to NTX treatment but were comparable to the Normal and DB-IN groups for at least 2 weeks after chronic exposure to 10(-3) to 10(-7) M NTX was terminated. No differences between Normal and DB-IN groups were noted. CONCLUSIONS: Topical application of NTX over a 10,000-fold range of dosage had no overt toxicity for the parameters studied, indicating that efficacy studies for the use of NTX in corneal wound healing are warranted. PMID- 17076635 TI - The effect of different grades of PLGA on characteristics of microspheres encapsulated with cyclosporine A. AB - The aim of this study was to evaluate the effect of different grades of poly D, L lactide-co-glycolide (PLGA) on the properties of microspheres encapsulated with Cyclosporine A (CyA). Microspheres were prepared by solvent evaporation method using three grades of PLGA. Various characteristics of microspheres such as morphology, size distribution, encapsulation efficiency and release profile were evaluated. Complementary studies were also carried out by Infrared (IR) spectroscopy and Differential scanning calorimetry (DSC) to evaluate possible drug-polymer interactions. Scanning electron microscopy (SEM) studies showed microspheres as spherical particles with CyA deposited as islands on the surface of spheres. Particle size range was 1-25 microm for microspheres made of PLGA (50:50) which showed the minimum size. Encapsulation efficiency was found to vary from 75% to 92% in various formulations. The profile of release was biphasic, showing an initial rapid phase followed by a continuous and slower rate thereafter. Microspheres made of grades 50:50 and 85:15 showed the highest and lowest amount of drug release, respectively. IR spectra for drug, polymer and microspheres did not indicate any chemical interaction between the components of microsphere and DSC thermograms revealed that CyA was present in its amorphous state within microspheres. In conclusion, the effect of polymer characteristics should be considered in microsphere formulations. In this study, suitable microspheres especially with PLGA (50:50) were prepared which allow the controlled release of CyA over a prolonged period of time. PMID- 17076634 TI - Copper and the prion protein: methods, structures, function, and disease. AB - The transmissible spongiform encephalopathies (TSEs) arise from conversion of the membrane-bound prion protein from PrP(C) to PrP(Sc). Examples of the TSEs include mad cow disease, chronic wasting disease in deer and elk, scrapie in goats and sheep, and kuru and Creutzfeldt-Jakob disease in humans. Although the precise function of PrP(C) in healthy tissues is not known, recent research demonstrates that it binds Cu(II) in an unusual and highly conserved region of the protein termed the octarepeat domain. This review describes recent connections between copper and PrP(C), with an emphasis on the electron paramagnetic resonance elucidation of the specific copper-binding sites, insights into PrP(C) function, and emerging connections between copper and prion disease. PMID- 17076636 TI - Controlled releases of FGF-2 and paclitaxel from chitosan hydrogels and their subsequent effects on wound repair, angiogenesis, and tumor growth. AB - A photocrosslinkable chitosan (Az-CH-LA) aqueous solution resulted in an insoluble hydrogel like a soft rubber within 30 sec of ultraviolet light (UV) irradiation. The photocrosslinked chitosan hydrogel showed strong sealing strength and potential use as a new tissue adhesive in surgical application. Paclitaxel, which is an anti-tumor reagent and a vascularization-inhibitor, retained in the photocrosslinked chitosan hydrogel, and were gradually released from the photocrosslinked chitosan hydrogel in vivo upon the degradation of the hydrogel. The paclitaxel-incorporated photocrosslinked chitosan hydrogels effectively inhibited tumor growth and angiogenesis in mice. On the other hand, the fibroblast growth factor (FGF)-2 molecules also retained in both the photocrosslinked chitosan and an injectable chitosan/IO(4)-heparin hydrogels, and were gradually released from the hydrogels upon their in vivo biodegradations. The activity of FGF-2 in the hydrogels was stable for long time (more than 14 days). The controlled release of biologically active FGF-2 molecules from the hydrogels caused an induction of the angiogenesis and, possibly, collateral circulation occurred in the healing-impaired diabetic (db/db) mice and the ischemic limbs of rats. The purpose of this review is to describe the effectiveness of the chitosan hydrogels (photocrosslinkable chitosan hydrogel and chitosan/IO(4)-heparin hydrogel) as a local drug delivery carrier for FGF-2 and paclitaxel to control wound repair, tumor growth, and angiogenesis. It is thus proposed that the chitosan hydrogels may be a promising new local carrier for drugs such as FGF-2 and paclitaxel. PMID- 17076637 TI - Chemical and biological factors in the control of Brucella and brucellosis. AB - Brucellosis is a highly contagious bacterial zoonosis that affects millions of people worldwide. Brucella is highly infectious, especially when aerosolized. The infection induces severe protracted diseases, which are both debilitating and incapacitating, hence, Brucella melitensis has been considered a potential biological warfare agent. In the battle against Brucella, it is crucial to know its chemical-structure and biochemistry-metabolic characteristics. It is well known that Brucella, as well as many other intracellular bacterial pathogens, has evolved to survive and even proliferate within monocytes and macrophages cells. Depending on the route of entry (complement, Fc, lectin or fibronectin receptors), the fate of the bacteria will vary; it may even segregate from the endocytic route towards the endoplasmic reticulum. This intracellular "non regular" behaviour of Brucella makes treatment difficult. Most antibiotics, although effective in vitro, do not actively pass through cellular membranes, or, once inside, may not reach the discrete intracellular niche where the bacteria is hidden. Therefore, complete eradication of the microorganisms is difficult to achieve, and the incidence of relapses is rather high. Taking these data into consideration, this review will evaluate the past, current and new trends in the control of brucellosis, paying special attention to the drug delivery systems as potential vectors for targeting these intracellular sites where the organisms are located. PMID- 17076638 TI - Controlled release calcium silicate based floating granular delivery system of ranitidine hydrochloride. AB - The objective of the present investigation was to prepare and evaluate floating granular delivery system consisting of (i) calcium silicate (CS) as porous carrier; (ii) ranitidine hydrochloride (RH), an anti-ulcer agent; and (iii) hydroxypropyl methylcellulose K4M (HPMC) and ethylcellulose (EC) as matrix forming polymers. The effect of various formulation and process variables on the particle morphology, particle size, micromeritic properties, percent drug content, in vitro floating behavior, and in vitro drug release from the floating granules was studied. The scanning electron microscopy (SEM) of granules revealed that that more pores of CS in secondary coated granules (SCG) were covered by the polymer film than those in primary coated granules (PCG). The formulation demonstrated favorable in vitro floating and drug release characteristics. The in vivo evaluation for the determination of pharmacokinetic parameters was performed in albino rats. Higher plasma concentration was maintained throughout the study period from the floating granules of RH. The enhanced bioavailability and elimination half-life observed in the present study may be due to the floating nature of the dosage form. The results suggested that CS is a useful carrier for the development of floating and sustained release preparations. PMID- 17076639 TI - Drug/cyclodextrin solid systems in the design of hydrophilic matrices: a strategy to modulate drug delivery rate. AB - The aim of this study was to investigate how the delivery rate of erodible sustained-release hydrophilic matrices intended for the delivery of poorly soluble drugs can be optimized through the incorporation of drug/cyclodextrin binary systems. Carvedilol (CAR), a sparingly water-soluble antihypertensive drug, was selected as a model for the study. As first, we attempted to improve CAR apparent solubility by association with hydroxypropyl-beta-cyclodextrin (HPbetaCD) and then incorporated CAR/HPbetaCD binary systems in sustained-release tablets made of poly(ethyleneoxide) (PEO). Solid CAR/HPbetaCD binary systems were prepared by physical mixing, kneading, co-melting and freeze-drying methods and characterized by DSC and X-ray powder diffractometry. The amount of CAR dissolved from all the HPbetaCD-containing systems was higher than pure CAR, the co-molten and freeze-dried products showing the best dissolution performance. The incorporation of the binary systems in PEO tablets resulted in a CAR release rate much higher than tablets containing only CAR. It was found that the time necessary to achieve complete release from the tablet was linearly related to the dissolution parameters of CAR/HPbetaCD powders. In the case of co-molten and freeze-dried products, all CAR content could be released in about 12 and 10 h, respectively. Our results demonstrate that the incorporation of drug/cyclodextrin solid systems in erodable PEO matrices intended for the delivery of poorly water soluble drugs is useful to modulate the release rate by controlling the dissolution properties of the drug inside the tablet. PMID- 17076640 TI - Particulate systems as adjuvants and carriers for peptide and protein antigens. AB - The most common feature for antigen-delivery systems is their particulate nature. Together with a certain depot effect, it is the particulate nature that primarily dictates whether the antigen-delivery system will be successful in inducing a certain type and strength of immune response. In this article, we will summarize recent data on particulate delivery systems for peptide and protein antigens with a main focus on lipid or polymer-based particles, all of which possess high potential as both preventive and therapeutic vaccines for parenteral, nasal, and possibly oral administration. PMID- 17076641 TI - Saponins from Quillaja saponaria Molina: isolation, characterization and ability to form immuno stimulatory complexes (ISCOMs). AB - ISCOMs have received much attention as vaccine adjuvants due to their immunostimulatory effects. They are colloidal particles typically comprised of phospholipids, cholesterol and Quil A, a crude mixture of saponins extracted from the bark of Quillaja saponaria Molina. We have previously shown that ISCOMs can be prepared by ether injection wherein an ether solution of phospholipids and cholesterol in a mass ratio of 5:2 is injected into a solution of Quil A at a mass ratio of 7 lipids: 3 Quil A. The aim of this study was firstly to isolate and characterise discrete fractions of Quil A and secondly to investigate which of these fractions were able to form ISCOMs by the method of ether injection. Six fractions of Quil A were isolated by semi-preparative reverse phase high performance liquid chromatography (RP-HPLC) and characterised by analytical HPLC, liquid chromatography tandem mass spectrometry (LC-MS) and the qualitative Liebermann-Burchard and Molisch tests for triterpenoids and carbohydrates respectively. ISCOMs were subsequently prepared from the isolated fractions by the method of ether injection and the resulting preparations characterized by photon correlation spectroscopy (PCS) and negative stain transmission electron microscopy (TEM). The molecular weights of the major compounds in the fractions ranged from approximately 1200 to approximately 2300 Da; all fractions tested positive for triterpenoids and saccharides and four of the fractions were identified as QS-7, QS-17, QS-18 and QS-21 by analysis (LC-MS and analytical HPLC). Injection of ether solutions of lipids into aqueous solutions of QS-17, QS 18 or QS-21 all resulted in homogeneous ISCOM dispersions. The combination of lipids and QS-7 by ether injection produced lamellae and liposomes as the prominent structures and a minor amount of ISCOMs. The remaining two hydrophilic, low molecular weight fractions of Quil A did not produce ISCOMs, instead liposomes and helical structures predominated in the samples. PMID- 17076642 TI - Isolation, characterization and study of enhancing effects on nasal absorption of insulin in rat of the total saponin from Acanthophyllum squarrosum. AB - OBJECTIVE: Isolation of the total saponins from Acanthophyllum squarrosum Boiss. and investigation of its surface activity, haemolytic effects on human erythrocytes as well as enhancing potentials on intranasal insulin absorption in rat in comparison with two other enhancers i.e. Quillaja total saponin (QTS) and sodium cholate (SC). MATERIALS AND METHODS: The decrease in blood glucose levels in five fasting rats following nasal administration of regular insulin solutions in the presence or absence of enhancers was determined by glucometric strips and used as an indication of insulin absorption. RESULTS: The results showed that ATS decreased surface tension of water to about 50 dyne.cm(-1) and caused complete haemolysis of human RBCs at a concentration of 250 microg.ml(-1). Following the instillation of solutions containing insulin and different absorption enhancers into the right nostril of rats, the percentage decrease in initial blood glucose was as follows: 72.46% (+/- 2.39%) for ATS, 63.22 % (+/-11.06%) for QTS and 60.06% (+/-14.93%) for SC. Percentage lowering in initial blood glucose concentrations against time showed that ATS exhibits a stronger effect than the two other enhancers although the difference was not statistically significant (p>0.05). CONCLUSION: ATS has a considerable absorption enhancing effect and can possibly be used to increase insulin bioavailability via nasal route. However the potential toxic effects of this saponin on nasal mucosa should be further evaluated. PMID- 17076643 TI - Artificial sun protection: sunscreens and their carrier systems. AB - The formulation of sunscreens products requires information about the properties of the substances responsible for the sun protection, their action mechanisms and the vehicles used for their administration. The improvement of the performance of sunscreen substances by a modification of their chemical, physical and technological properties or by the use of conventional and novel carriers is described in this review. The sun protection achieved by the use of sunscreens incorporated into carriers in terms of the sun protection factor (SPF) and other performance indices is also shown. Current manufacturing methods to incorporate sunscreen substances into vehicles are briefly described. Furthermore, basic information related to the dermatological effects of the UV radiation classified according to the different spectral regions of the UV radiation is presented in this manuscript. PMID- 17076644 TI - Topical delivery of aceclofenac from lecithin organogels: preformulation study. AB - The purpose of this research is to evaluate the suitability of lecithin organogels containing aceclofenac for topical application. The present article focuses on the preformulation part of the whole research work. Thin layer chromatography was carried out to determine lecithin's purity. The excipients for formulating lecithin organogel were screened. Lecithin organogels are thermo reversible in nature and hence gelation temperature study was carried out to determine the temperature where Sol-Gel and Gel-Sol transformation takes place. Partition coefficient of the drug was estimated. Drug solubility in plain oil and organogel containing reverse micelles was estimated. Effect of water added on the properties of lecithin organogels such as X-ray diffraction pattern, conductivity and viscosity were determined. Microscopy of the gel sample has been carried out at different magnifications. The pseudo ternary phase diagram has been constructed to determine the organogel existence region. The permeation study of aceclofenac from different concentrations of lecithin organogels [200 mM, 300 mM and 400 mM] has been determined using cellulose acetate membrane (0.45 micro) and excised rat skin. Lecithin organogel in ethyl oleate has desired stability and consistency. A single spot on the TLC plate confirms the purity of soy lecithin to be used in organogel formation. Aceclofenac solubility was found to be more in lecithin/oil reverse micellar system as compared to its solubility in oil. The X ray diffraction pattern confirms the incorporation of water in micellar gel network. The physical properties of organogels are affected by water incorporated and concentration of gelator. The permeation of aceclofenac through artificial membrane and excised rat skin demonstrated the same trend and were in the following order 200 mM>300 mM>400 mM. The results showed that organogel exhibits useful pharmaceutical properties. PMID- 17076645 TI - Multiple emulsions: an overview. AB - Multiple emulsions are complex polydispersed systems where both oil in water and water in oil emulsion exists simultaneously which are stabilized by lipophillic and hydrophilic surfactants respectively. The ratio of these surfactants is important in achieving stable multiple emulsions. Among water-in-oil-in-water (w/o/w) and oil-in-water-in-oil (o/w/o) type multiple emulsions, the former has wider areas of application and hence are studied in great detail. Formulation, preparation techniques and in vitro characterization methods for multiple emulsions are reviewed. Various factors affecting the stability of multiple emulsions and the stabilization approaches with specific reference to w/o/w type multiple emulsions are discussed in detail. Favorable drug release mechanisms and/or rate along with in vivo fate of multiple emulsions make them a versatile carrier. It finds wide range of applications in controlled or sustained drug delivery, targeted delivery, taste masking, bioavailability enhancement, enzyme immobilization, etc. Multiple emulsions have also been employed as intermediate step in the microencapsulation process and are the systems of increasing interest for the oral delivery of hydrophilic drugs, which are unstable in gastrointestinal tract like proteins and peptides. With the advancement in techniques for preparation, stabilization and rheological characterization of multiple emulsions, it will be able to provide a novel carrier system for drugs, cosmetics and pharmaceutical agents. In this review, emphasis is laid down on formulation, stabilization techniques and potential applications of multiple emulsion system. PMID- 17076646 TI - New perspectives in the treatment of hematological malignancies. PMID- 17076647 TI - DNA hypermethylation of myeloid cells, a novel therapeutic target in MDS and AML. AB - Differential methylation of CpG islands is a regulatory mechanism for promoter activity of different classes of genes, including tissue-specific genes. These CpG islands are targets for transformation-associated, aberrant hypermethylation activity during leukemogenesis. Therefore the pharmacological reversion of this methylator phenotype (e.g. by reactivation of tumor suppressor gene expression) is an important rationale for development of inhibitors of DNA methyltransferase activity. In vitro, inhibition of methylation using azanucleosides results in modest differentiation of transformed myeloid cell lines. In vivo, low doses of these agents induce DNA demethylation of malignant myeloid cells. Indeed, the first drug specifically approved for the treatment of myelodysplastic syndrome (MDS) was the azanucleoside 5-azacytidine (Vidaza). The most potent DNA demethylating agent available, 5-aza-2' deoxycytidine (Decitabine, Dacogen) also has recently been approved by the U.S.A. FDA for treatment of MDS of all subtypes. About 30 % of MDS patients with an abnormal karyotype have normalization of their karyotype after receiving the drug. This activity is especially relevant in patients with high-risk karyotypic abnormalities (complex karyotype and/or abnormalities of chromosome 7) compared to patients with intermediate-risk karyotype. Both drugs offer a novel, non-intensive therapeutic approach, particularly in the older patient population who due to comorbidities and/or reduced performance status are ineligible for aggressive chemotherapies. Target genes being particularly prone to demethylation by these drugs in the aberrant cells (e.g. p15/INK4b) are under active investigation. Future translational and clinical studies will be aimed at improving the response rate and duration of response to non-intensive treatment with demethylating agents, by studying rational drug combinations e.g. with inhibitors of histone deacetylase activity. PMID- 17076648 TI - Induction of apoptosis via the modulation of reactive oxygen species (ROS) production in the treatment of myeloid leukemia. AB - Recent advances in genetic and molecular biology have provided greater insight into the biology of acute myeloid leukemia (AML). These investigations have shown that AML is a heterogeneous disease of biologically different entities. Current therapeutic approaches to AML are based on chemotherapy, but the side effects of the drugs used and various complications, including infections and bleeding, are sometimes fatal. In addition, responses to therapy and long-term outcome differ depending on the subentity in question. Therefore, it is essential to develop new therapeutic strategies such as biology adapted treatment based on the individual molecular pathogenesis of AML. Natural compounds appear to be safer than the current chemotherapeutic drugs, and we have therefore sought new potential agents among various natural compounds with the ability to induce the apoptosis of myeloid leukemic cells. Recently, we found that a highly toxic reactive oxygen species (ROS) generated via the hydrogen peroxide/myeloperoxidase [H(2)O(2)/MPO/halide] system by natural compounds induces apoptosis in MPO positive leukemic cells. This result is of great interest in establishing novel therapeutic approaches to AML mediated through ROS-induced apoptosis of leukemic cells. PMID- 17076649 TI - Developing target therapy against oncogenic tyrosine kinase in myeloid maliganacies. AB - Myeloid malignancies are frequently associated with translocations and mutations of tyrosine kinase genes. Fusion genes involving ABL, ARG, PDGFRs, JAK2, SYK, TRKC, and FGFRs, and gain-of-function mutations of FLT3, KIT and JAK2 have been detected at various rates in myeloproliferative disease and acute myeloid leukemia. Furthermore, abnormal overexpression of tyrosine kinases such as FLT3 has also been reported. These gene products are constitutively activated and potentially transform hematopoietic cells by augmentation of proliferation and enhanced viability. Since the fusion or mutation of tyrosine kinase is a primary and central event in chronic myeloproliferative diseases, targeting the kinase activity has been thought to be an ideal intervention to treat these diseases. The clinical success of imatinib for chronic myeloid leukemia has made this idea a reality, and has accelerated the development of new tyrosine kinase inhibitors (TKIs). Challenging studies with TKIs have also been reported for acute myeloid leukemia. This review will focus on recent trials of TKIs against oncogenic tyrosine kinases (ABL, PDGFRs, FLT3 and KIT) in myeloid malignancies. PMID- 17076650 TI - Immunomodulatory drugs (IMiDs): a new treatment option for myelodysplastic syndromes. AB - The IMiDs represent a new proprietary class of thalidomide analogues that possess greater potency and less toxicity than the parent compound. As a group, these agents share the pharmacologic property of modulating cellular response to ligand activation, the precise biologic effect of which is cell lineage and stimulant dependent. Lenalidomide (CC-5013; Revlimid), a second generation IMiD, has shown significant erythropoietic activity in patients with lower risk MDS that have failed or are not candidates for recombinant erythropoietin treatment. Unlike cytokine therapy, lenalidomide suppresses select MDS clones and enhances erythropoietin receptor signaling to restore erythropoiesis. Activity is greatest in patients with interstitial deletions involving chromosome 5q31.1. A multicenter phase II study reported a 76 % overall transfusion response rate in transfusion-dependent patients with deletion 5q, with 67 % achieving transfusion independence after a median interval of 4.6 weeks of treatment. Cytogenetic responses were observed in 73% of patients with complete cytogenetic remission in 45% patients. Both transfusion response and cytogenetic response frequency were independent of karyotype complexity, raising excitement that this new treatment strategy might favorably alter the natural history of disease in higher risk patients with deletion 5q. Lenalidomide was approved by the U.S. Food and Drug Administration on December 27, 2005, for the treatment of IPSS Low and intermediate-1 risk MDS patients with del(5q) abnormality. A phase III Intergroup trial (ECOG 2905) will test the capacity to potentiate erythropoietin response by comparing response to lenalidomide monotherapy to the combination of darbepoetin and lenalidomide in non-deletion 5q MDS patients. PMID- 17076651 TI - Monoclonal antibodies for the treatment of acute myeloid leukemia. AB - Currently, patients with acute myeloid leukemia (AML) are treated with cytotoxic chemotherapy and hematopoietic stem cell transplantation (HSCT). With this approach, the majority of patients still die of their disease because of both treatment-related mortality and relapse. Recently, monoclonal antibodies and immunoconjugates have been developed which potentially may increase the efficacy of treatment and decrease morbidity and mortality by specifically targeting the malignant cell. Unconjugated monoclonal antibodies have shown only moderate activity. A second, more effective, approach involves antibody conjugation with radioactive particles or chemotherapeutic agents, such as, immunotoxins, targeted delivery of cell killing. The antigens CD33, CD45, and CD66, are three antigens to which monoclonal antibodies have been directed. Most experience has been with gemtuzumab ozogamicin (Mylotarg) which is an immunoconjugate of an anti-CD33 antibody chemically linked to a potent cytotoxic agent, calicheamicin. Gemtuzumab ozogamicin appears to be particularly active in patients with acute promyelocytic leukemia, possibly related to the high expression of the CD33 antigen on the cell surface. Although gemtuzumab ozogamicin has activity as a single agent, the most promising result may be seen when this agent is combined with conventional cytotoxic chemotherapy. Preliminary studies have suggested a high complete remission rate and randomized clinical trials are underway. A unique potential toxicity has been identified, namely venoocclusive disease or sinusoidal obstructive syndrome which may be problematic among patients who subsequently undergo HSCT. An additional strategy includes radiolabeled monoclonal antibodies to intensify the conditioning regimen prior to HSCT. The most promising results have been obtained with radiolabeled anti-CD45 antibodies. PMID- 17076652 TI - New tyrosine kinase inhibitors in the treatment of chronic myeloid leukemia. AB - Imatinib mesylate, Abl tyrosine kinase inhibitor, has improved the treatment of Bcr-Abl-positive leukemia such as chronic myeloid leukemia (CML) and Philadelphia chromosome positive acute lymphoblastic leukemia (Ph(+)ALL). However, resistance is often reported in patients with advanced-stage disease. Several novel tyrosine kinase inhibitors, which have been developed to override imatinib resistance mechanisms such as overexpression of Bcr-Abl and point mutations within the Abl kinase domain, are currently competing. Inhibitors of Abl tyrosine kinase are divided into two main groups, namely, ATP-competitive and ATP non-competitive inhibitors. Moreover, ATP-competitive inhibitors are fall into two subclasses, i.e. the Src/Abl inhibitors, and 2-phenylaminopyrimidin-based compounds. Dasatinib (formerly BMS-354825), AP23464, SKI-606 and PD166326 are classified as Src/Abl inhibitors while AMN107 and NS-187 (INNO-406) belong to the latter subclass of inhibitors. Among these agents, clinical studies on dasatinib and AMN107 had started earlier than the others and favorable results are accumulating. Clinical studies of other compounds including NS-187 (INNO-406) will be performed in rapid succession. Because of its strong affinity, most ATP competitive inhibitors may be effective against imatinib-resistant patients. However, to date, an ATP-competitive inhibitor that can inhibit the phosphorylation of T315I Bcr-Abl has not yet been developed. To address this problem, ATP non-competitive inhibitors such as ON012380, Aurora kinase inhibitor VX-680 and p38 MAP kinase inhibitor BIRB-796 have been developed. It may be necessary for the improvement of CML and Ph(+)ALL treatment to be taken into consideration of the combination therapy with novel ATP-competitive inhibitors and these agents. PMID- 17076653 TI - Recent advances in the treatment of Multiple Myeloma. AB - Multiple Myeloma (MM) remains an incurable plasma cell malignancy in the bone marrow (BM) despite conventional therapies as well as high-dose therapies with stem cell support. Therefore novel biologically-based therapeutic approaches are required. Recently, intensive laboratory and preclinical studies have identified and validated therapeutic molecular targets in MM. In particular, recognition of the biologic significance of the BM microenvironment in MM pathogenesis and as a potential target for novel therapeutics has derived several promising approaches. Novel FDA approved agents including thalidomide/thalomid, its immunomodulatory derivatives lenalidomide/Revlimid, and proteasome inhibitor bortezomib/Velcade are directed at molecular targets not only in MM cells but also in its BM milieu, and have already achieved promising results in clinical studies. Here we discuss the mechanisms of action of these novel drugs and their clinical application, alone or combined with conventional or novel drugs. PMID- 17076654 TI - Disease modifying therapy for AD? AB - Alzheimer's disease (AD) is the most common form of dementia in industrialized nations. If more effective therapies are not developed that either prevent AD or block progression of the disease in its very early stages, the economic and societal cost of caring for AD patients will be devastating. Only two types of drugs are currently approved for the treatment of AD: inhibitors of acetyl cholinesterase, which symptomatically enhance cognitive state to some degree but are not disease modifying; and the adamantane derivative, memantine. Memantine preferentially blocks excessive NMDA receptor activity without disrupting normal receptor activity and is thought to be a neuroprotective agent that blocks excitotoxicty. Memantine therefore may have a potentially disease modifying effect in multiple neurodegenerative conditions. An improved understanding of the pathogeneses of AD has now led to the identification of numerous therapeutic targets designed to alter amyloid beta protein (Abeta) or tau accumulation. Therapies that alter Abeta and tau through these various targets are likely to have significant disease modifying effects. Many of these targets have been validated in proof of concept studies in preclinical animal models, and some potentially disease modifying therapies targeting Abeta or tau are being tested in the clinic. This review will highlight both the promise of and the obstacles to developing such disease modifying AD therapies. PMID- 17076655 TI - Alois Alzheimer and Alzheimer's disease: a centennial perspective. AB - The year 2006 is the centenary of the famous presentation of Alois Alzheimer which first described the neuropathology of Alzheimer's disease (AD). Since this presentation, enormous progress has been made in understanding the biology of AD. The central role of the beta-amyloid protein (Abeta) in the pathogenesis of AD and the relationship between plaque and tangle pathology is now much better understood. In this article, we review the current status of the amyloid hypothesis of AD and its role in the development of future therapy. PMID- 17076656 TI - Growth hormone releasing peptide-6 acts as a survival factor in glutamate-induced excitotoxicity. AB - Chronic systemic treatment given to adult male rats with growth hormone releasing peptide-6, an agonist of the ghrelin receptor, increases insulin-like growth factor I levels in various brain regions, including the hypothalamus and cerebellum. Furthermore, intracellular signalling cascades normally associated with anti-apoptotic actions are activated in the same areas and are coincident with decreased basal cell death. Because abnormally high concentrations of glutamate can lead to overexcitation of neurones leading to cell damage and/or death, we investigated whether administration of growth hormone releasing peptide 6 attenuates monosodium glutamate-induced apoptosis in the rat hypothalamus and cerebellum. Glutamate increased activation of caspase 9 followed by cleavage of caspase 7, which in turn fragmented poly(ADP-ribose) polymerase, terminating in cell death in both the hypothalamus and cerebellum. Growth hormone releasing peptide-6 reversed glutamate-induced cell death by decreasing activation of caspases 9 and 7 and poly(ADP-ribose) polymerase fragmentation. These results provide a better understanding of the neuroprotective role of growth hormone secretagogues and the mechanisms involved. PMID- 17076657 TI - Recombinant human granulocyte colony-stimulating factor protects against MPTP induced dopaminergic cell death in mice by altering Bcl-2/Bax expression levels. AB - Granulocyte colony-stimulating factor (G-CSF) has been used for the treatment of neutropenia in hematologic disorders. The neuroprotective effects of G-CSF were reported in neurological disease models. In the present study, we examined whether G-CSF can protect dopaminergic neurons against MPTP-induced cell death in a mouse model of Parkinson's disease. Mice of one group were injected intraperitoneally with MPTP for five consecutive days, those of another group with MPTP and intraperitoneal G-CSF at 2 days and 1 day before the first MPTP injection, and 30 min before each MPTP injection, while control mice received saline injections. Immunohistochemistry, western blotting analysis, and HPLC were performed to evaluate damage of substantia nigra dopaminergic neurons and expression of Bcl-2 and Bax protein. MPTP induced dopaminergic cell death in the substantia nigra. G-CSF significantly prevented MPTP-induced loss of tyrosine hydroxylase-positive neurons (p < 0.05), increased Bcl-2 protein and decreased Bax protein expression. Our findings indicate that G-CSF provides neuroprotection against MPTP-induced cell death and this effect is mediated by increasing Bcl-2 expression levels and decreasing Bax expression levels in C57BL/6 mice. PMID- 17076658 TI - Uracil nucleotides stimulate human neural precursor cell proliferation and dopaminergic differentiation: involvement of MEK/ERK signalling. AB - Isolation and propagation of neural stem cells derived from human brain tissue uniquely enables the study of human neurogenesis in vitro. In addition, ex vivo expanded human neural stem/precursor cells (NPCs) may offer novel therapeutic strategies. We investigated the effects of extracellular nucleotides on the proliferation and differentiation of human mesencephalic neural stem/precursor cells (hmNPCs). When combined with the mitogens epidermal growth factor and fibroblast growth factor 2, UTP (1 microm) boosted proliferation of hmNPCs as shown by increased expression of the proliferation marker proliferating cell nuclear antigen (330%). UTP-induced proliferation was abrogated by the preferential P2Y receptor blocker pyridoxal phosphate-6-azophenyl-2',4' disulphonic acid (PPADS). UTP also stimulated dopaminergic differentiation. Treatment with UTP (100 microm) increased the number of tyrosine hydroxylase (TH) positive cells and TH protein by 267 and 319% respectively. UTP-stimulated dopaminergic differentiation of hmNPCs was blocked by the P2 receptor antagonists suramin (10 microm) and PPADS (100 microm). In addition, UDP (1 microm) enhanced TH protein expression by 194%. During differentiation, treatment with UTP stimulated the extracellular signal-regulated kinase (ERK) pathway. Both ERK1/2 phosphorylation and dopaminergic differentiation were inhibited by U0126, a selective ERK kinase inhibitor, as well as by suramin. When other P2 receptor agonists (ATP, ADP and adenosine 5'-O-(2-thiophosphate) (ADPbetaS); all 100 microm) were applied, both proliferation and dopaminergic differentiation of NPCs were compromised. We conclude that uracil nucleotides exert specific P2 receptor mediated effects on midbrain-derived human NPCs, and may be used to enhance both proliferation and dopaminergic differentiation. PMID- 17076659 TI - Glutamate-stimulated ATP release from spinal cord astrocytes is potentiated by substance P. AB - ATP has recently emerged as a key molecule mediating pathological pain. The aim of this study was to examine whether spinal cord astrocytes could be a source of ATP in response to the nociceptive neurotransmitters glutamate and substance P. Glutamate stimulated ATP release from these astrocytes and this release was greatly potentiated by substance P, even though substance P alone did not elicit ATP release. Substance P also potentiated glutamate-induced inward currents, but did not cause such currents alone. When glutamate was applied alone it acted exclusively through alpha-amino-3-hydroxy-5-methylisoxazole-4-proprionate receptors to stimulate Ca(2+) influx-dependent ATP release. However, when substance P was co-applied with glutamate, ATP release could be elicited by activation of NMDA and metabotropic glutamate receptors. Activation of neurokinin receptor subtypes, protein kinase C and phospholipases A(2), C and D were needed for substance P to bring about its effects. These results suggest that astrocytes may be a major source of ATP in the spinal cord on activation of nerve fibres that release substance P and glutamate. PMID- 17076660 TI - Neuroprotectant FK506 inhibits glutamate-induced apoptosis of astrocytes in vitro and in vivo. AB - Neuron-astrocyte interactions are critical for signalling, energy metabolism, extracellular ion and glutamate homeostasis, volume regulation and neuroprotection in the CNS. Glutamate uptake by astrocytes may prevent excitotoxic glutamate elevation and determine neuronal survival. However, an excess of glutamate can cause the death of astrocytes. FK506, an inhibitor of calcineurin, and an immunosuppressive drug, is neuroprotective in animal models of neurologic diseases, including focal and global ischaemia. In the present work, we demonstrate that a single injection of FK506 60 min after a transient middle cerebral artery occlusion (MCAo) significantly decreases the number of terminal deoxynucleotidyl transferase nick-end labelling (TUNEL)-positive cells in the ischaemic cortex and striatum. Using 3-D confocal microscopy we found that, 24 h after MCAo, many TUNEL-positive cells in the ischaemic striatum and cortex are astrocytes. Furthermore, we demonstrate that exposure of cultured cortical astrocytes to 50-100 mM Glu for 24 h induces apoptotic alterations in nuclear morphology, DNA fragmentation, dissipation of mitochondrial transmembrane potential (DeltaPsi) and caspase activation. FK506 (1 muM) efficiently inhibits Glu-induced apoptosis of cultured astrocytes, DNA fragmentation and changes in mitochondrial DeltaPsi. Our findings suggest that modulation of glutamate-induced astrocyte death early after reperfusion may be a novel mechanism of FK506 mediated neuroprotection in ischaemia. PMID- 17076661 TI - TAp73 isoforms antagonize Notch signalling in SH-SY5Y neuroblastomas and in primary neurones. AB - p73, like Notch, has been implicated in neurodevelopment and in the maintenance of the mature central nervous system. In this study, by the use of reporter-gene assays, we demonstrate that C-promoter binding factor-1 (CBF-1)-dependent gene transcription driven by the Notch-1 intracellular domain (N1(ICD)) is potently antagonized by exogenously expressed transactivating (TA) p73 splice variants in SH-SY5Y neuroblastomas and in primary neurones. Time course analysis indicated that the inhibitory effects of TAp73 are direct and are not mediated via the product of a downstream target gene. We found that endogenous TAp73 stabilized by either c-Abl or cisplatin treatment also potently antagonized N1(ICD)/CBF-1 dependent gene transcription. Furthermore, western blotting revealed that exogenous TAp73 suppressed endogenous hairy and enhancer of split-1 (HES-1) protein levels and antagonized the increase in HES-1 protein induced by exogenous N1(ICD) expression. Evidence of a direct physical interaction between N1(ICD) and TAp73alpha was demonstrated by co-immunoprecipitation. Using Notch deletion constructs, we demonstrate that TAp73alpha binds the N1(ICD) in a region C terminal of aa 2094. Interestingly, DeltaNp73alpha and TAp73alpha(R292H) also co purified with N1(ICD), but neither inhibited N1(ICD)/CBF-1-dependent transcription. This suggests that an intact transactivation (TA) domain and the ability to bind DNA are necessary for TAp73 to antagonize Notch signalling. Finally we found that TAp73alpha reversed the N1(ICD)-mediated repression of retinoic acid-induced differentiation of SH-SY5Y neuroblastomas, providing functional evidence for an inhibitory effect of TAp73alpha on notch signalling. Collectively, these findings may have ramifications for neurodevelopment, neurodegeneration and oncogenesis. PMID- 17076662 TI - Mice deficient for galanin receptor 2 have decreased neurite outgrowth from adult sensory neurons and impaired pain-like behaviour. AB - Expression of the neuropeptide galanin is markedly up-regulated within the adult dorsal root ganglia (DRG) following peripheral nerve injury. We have previously demonstrated that galanin knockout (Gal-KO) mice have a developmental loss of a subset of DRG neurons. Galanin also plays a trophic role in the adult animal, and the rate of peripheral nerve regeneration and neurite outgrowth is reduced in adult Gal-KO mice. Here we describe the characterization of mice with an absence of GalR2 gene transcription (GalR2-MUT) and demonstrate that they have a 15% decrease in the number of calcitonin gene-related peptide (CGRP) expressing neuronal profiles in the adult DRG, associated with marked deficits in neuropathic and inflammatory pain behaviours. Adult GalR2-MUT animals also have a one third reduction in neurite outgrowth from cultured DRG neurons that cannot be rescued by either galanin or a high-affinity GalR2/3 agonist. Galanin activates extracellular signal-regulated kinase (ERK) and Akt in adult wild-type (WT) mouse DRG. Intact adult DRG from GalR2-MUT animals have lower levels of pERK and higher levels of pAkt than are found in WT controls. These data suggest that a lack of GalR2 activation in Gal-KO and GalR2-MUT animals is responsible for the observed developmental deficits in the DRG, and the decrease in neurite outgrowth in the adult. PMID- 17076663 TI - Heterochromatin-mediated control of virulence gene expression. AB - In recent years, the sequencing and annotation of complete genomes, together with the development of genetic and proteomic techniques to study previously intractable eukaryotic microbes, has revealed interesting new themes in the control of virulence gene expression. Families of variantly expressed genes are found adjacent to telomeres in the genomes of both pathogenic and non-pathogenic organisms. This subtelomeric DNA is normally heterochromatic and higher-order chromatin structure has now come to be recognized as an important factor controlling both the evolution and expression dynamics of these multigene families. In eukaryotic cells, higher-order chromatin structure plays a central role in many DNA processes including the control of chromosome integrity and recombination, DNA partitioning during cell division, and transcriptional control. DNA can be packaged in two distinct forms: euchromatin is relatively accessible to DNA binding proteins and generally contains active genes, while heterochromatin is densely packaged, relatively inaccessible and usually transcriptionally silent. These features of chromatin are epigenetically inherited from cell cycle to cell cycle. This review will focus on the epigenetic mechanisms used to control expression of virulence genes in medically important microbial pathogens. Examples of such control have now been reported in several evolutionarily distant species, revealing what may be a common strategy used to regulate many very different families of genes. PMID- 17076664 TI - Bacillus sphaericus mosquitocidal toxin (MTX) and pierisin: the enigmatic offspring from the family of ADP-ribosyltransferases. AB - The mosquitocidal toxin (MTX) from Bacillus sphaericus and the apoptosis-inducing pierisin-1 from the cabbage butterfly Pieris rapae are two of the most intriguing members of the family of ADP-ribosyltransferases. They are both approximately 100 kDa proteins, composed of an N-terminal ADP-ribosyltransferase (approximately 27 kDa) and a C-terminal putative binding and translocation domain (approximately 70 kDa) consisting of four ricin-B-like domains. While they both share structural homologies, with an overall amino acid sequence identity of approximately 30% that becomes approximately 50% at the level of the catalytic core, and functional similarities, notably in terms of enzyme regulation, they seem to largely differ with regard to their targets or cell internalization mechanisms. MTX ADP ribosylates numerous proteins in lysates of target insect cells at arginine residues, whereas pierisin-1 modifies DNA of insect and mammalian cells by ADP ribosylation at 2'-deoxyguanosine residues resulting in DNA adducts, mutations and eventually apoptosis. This target specificity differentiates pierisin-1 from all other ADP-ribosyltransferases described so far, and implies that the enzyme must reach the nucleus of target cells. The recently solved crystal structure of MTX catalytic domain is helpful to reveal new insights into structural organization, catalytic mechanisms, proteolytic activation and autoinhibition of both enzymes. The uptake and processing of the ADP-ribosyltransferases is discussed. PMID- 17076665 TI - A specific secretion system mediates PPE41 transport in pathogenic mycobacteria. AB - Mycobacterial genomes contain two unique gene families, the so-called PE and PPE gene families, which are highly expanded in the pathogenic members of this genus. Here we report that one of the PPE proteins, i.e. PPE41, is secreted by pathogenic mycobacteria, both in culture and in infected macrophages. As PPE41 lacks a signal sequence a dedicated secretion system must be involved. A single gene was identified in Mycobacterium marinum that showed strongly reduced PPE41 secretion. This gene was located in a gene cluster whose predicted proteins encode components of an ESAT-6-like secretion system. This cluster, designated ESX-5, is conserved in various pathogenic mycobacteria, but not in the saprophytic species Mycobacterium smegmatis. Therefore, different regions of this cluster were introduced in M. smegmatis. Only introduction of the complete ESX-5 locus resulted in efficient secretion of heterologously expressed PPE41. This PPE secretion system is also involved in the virulence of pathogenic mycobacteria, as the ESX-5 mutant of M. marinum was affected in spreading to uninfected macrophages. PMID- 17076666 TI - Excision and transfer of the Mesorhizobium loti R7A symbiosis island requires an integrase IntS, a novel recombination directionality factor RdfS, and a putative relaxase RlxS. AB - The Mesorhizobium loti strain R7A symbiosis island is an Integrative Conjugative Element (ICE), herein termed ICEMlSymR7A, which integrates into a phetRNA gene. Integration reconstructs the phetRNA gene at one junction with the core chromosome, and a direct repeat of the 3-prime 17 bp of the gene is formed at the other junction. We show that the ICEMlSymR7AintS gene, which encodes an integrase of the phage P4 family, is required for integration and excision of the island. Excision also depended on a novel recombination directionality factor encoded by msi109 (rdfS). Constitutive expression of rdfS resulted in curing of ICEMlSymR7A. The rdfS gene is part of an operon with genes required for conjugative transfer, allowing co-ordinate regulation of ICEMlSymR7A excision and transfer. The excised form of ICEMlSymR7A was detectable during exponential growth but occurred at higher frequency during stationary phase. ICEMlSymR7A encodes homologues of the traR and traI genes of Agrobacterium tumefaciens that regulate Ti plasmid transfer via quorum sensing. The presence of a plasmid with cloned island traR traI2 genes resulted in excision of ICEMlSymR7A in all cells regardless of culture density, indicating that excision may be similarly regulated. Maintenance of ICEMlSymR7A in these cells depended on msi106 (rlxS) that encodes a putative relaxase. Transfer of the island to non-symbiotic mesorhizobia required intS, rlxS and rdfS. The rdfS and rlxS genes are conserved across a diverse range of alpha-, beta- and gamma-proteobacteria and identify a large family of genomic islands with a common transfer mechanism. PMID- 17076667 TI - Complete alanine scanning of the two-component lantibiotic lacticin 3147: generating a blueprint for rational drug design. AB - Lantibiotics are post-translationally modified antimicrobial peptides which are active at nanomolar concentrations. Some lantibiotics have been shown to function by targeting lipid II, the essential precursor of cell wall biosynthesis. Given that lantibiotics are ribosomally synthesized and amenable to site-directed mutagenesis, they have the potential to serve as biological templates for the production of novel peptides with improved functionalities. However, if a rational approach to novel lantibiotic design is to be adopted, an appreciation of the roles of each individual amino acid (and each domain) is required. To date no lantibiotic has been subjected to such rigorous analysis. To address this issue we have carried out complete scanning mutagenesis of each of the 59 amino acids in lacticin 3147, a two-component lantibiotic which acts through the synergistic activity of the peptides LtnA1 (30 amino acids) and LtnA2 (29 amino acids). All mutations were performed in situ in the native 60 kb plasmid, pMRC01. A number of mutations resulted in the elimination of detectable bioactivity and seem to represent an invariable core within these and related peptides. Significantly however, of the 59 amino acids, at least 36 can be changed without resulting in a complete loss of activity. Many of these are clustered to form variable domains within the peptides. The information generated in this study represents a blue-print that will be critical for the rational design of lantibiotic-based antimicrobial compounds. PMID- 17076668 TI - A novel oxalosuccinate-forming enzyme involved in the reductive carboxylation of 2-oxoglutarate in Hydrogenobacter thermophilus TK-6. AB - We have previously demonstrated that the reductive carboxylation of 2 oxoglutarate in Hydrogenobacter thermophilus TK-6 is not simply a reversal of the oxidative decarboxylation catalysed by isocitrate dehydrogenase (ICDH). The reaction involves a novel biotin protein (carboxylating factor for ICDH-CFI) and ATP. In this study, we have analysed the ICDH/CFI system responsible for the carboxylation reaction. Sequence analysis revealed a close relationship between CFI and pyruvate carboxylase. Rather unexpectedly, the rate of ATP hydrolysis was greater than that of isocitrate formation or NADH oxidation. Furthermore, ATP hydrolysis catalysed by CFI was dependent on 2-oxoglutarate but not on ICDH, suggesting that a carboxylated product is formed in the absence of ICDH. The product, which was detectable only at low temperatures, was identified as oxalosuccinate. Thus, CFI was confirmed to be a novel enzyme that catalyses the carboxylation of 2-oxoglutarate to form oxalosuccinate, which corresponds to the first step of the reductive carboxylation from 2-oxoglutarate to isocitrate. The CFI-ICDH system may also be present in mammals, where it could play a significant role in modulating central metabolism. PMID- 17076669 TI - nilR is necessary for co-ordinate repression of Xenorhabdus nematophila mutualism genes. AB - The bacterial mutualist Xenorhabdus nematophila colonizes a specific region of its nematode host Steinernema carpocapsae. We previously reported the identification of a chromosomal locus encoding three X. nematophila genes of unknown function, nilA, B and C, that are each necessary for colonization. Subsequent work indicated the global regulator Lrp is a repressor of nilC: nilC transcription is elevated in an lrp mutant and Lrp interacts directly with the nilC promoter. In this manuscript, we report the identification of an additional gene, nilR, required for repression of nilC transcription. We show that nilR and lrp mutants also have elevated expression of nilA and nilB, demonstrating that nilA, B and C are co-ordinately regulated. nil gene expression is derepressed most strongly when both nilR and lrp are lacking, suggesting NilR and Lrp synergistically repress nil transcription. NilR contains a helix-turn-helix-type DNA binding domain and likely acts directly at promoters. A comparison of the wild type and nilR proteomes indicates that NilR, unlike Lrp, regulates a small number of genes. Finally, X. nematophila carrying an ectopic copy of nilR colonizes at approximately 60-fold lower levels than the control strain, suggesting that derepression of nil gene expression is necessary for nematode colonization. PMID- 17076670 TI - The inflammation-associated Salmonella SopA is a HECT-like E3 ubiquitin ligase. AB - Salmonella translocate a group of type III effectors into the host cells to induce entry, promote survival and cause intestinal inflammation. Although the biochemical and cellular mechanisms of how bacterial effectors function inside host cells remain largely unknown, studies have indicated that a likely strategy is to exploit host cellular pathways through functional mimicry. We report here that SopA, a Salmonella type III effector, mimics the mammalian HECT E3 ubiquitin ligase. SopA preferentially uses the host UbcH5a, UbcH5c and UbcH7 as E2s, which are involved in inflammation. Both the wild-type SopA and the mutant SopAC753S were expressed and translocated at similar levels during the infection of HeLa cells. A Salmonella strain expressing a catalytically incompetent SopAC753S mutant had reduced Salmonella-induced polymorphonuclear leukocytes transepithelial migration. We speculate that SopA ubiquitinate bacterial/host proteins involved in Salmonella-induced intestinal inflammation. PMID- 17076671 TI - Inhibition of term decidual NK cell cytotoxicity by soluble HLA-G1. AB - OBJECTIVES: Soluble (s)HLA-G1 is produced by trophoblast cells. Aim was to analyze the capacities and mechanisms of sHLA-G1 to regulate interleukin (IL)-2 induced cytotoxicity of natural killer (NK) cells from human deciduas. METHODS: Natural killer cells were isolated from decidual layers of term placentae, stimulated or not with IL-2 and supplemented with various concentrations of recombinant soluble HLA-G1 (sHLA-G1). For NK cell cytotoxicity assays, K562 cells were used as targets. Expression of signal transducer and activator of transcription 3 (STAT3) and perforin was analyzed by Western blotting. Apoptosis was examined by assessment of poly(ADP-ribose) polymerase cleavage. NK cells were analyzed by flow cytometry for IL-2receptor-alpha (IL-2R alpha; CD25) and transferrin receptor CD71 expression. RESULTS: Interleukin-2 increases CD71, STAT3, perforin expression and cytotoxic potential of NK cells. Expression of CD71, STAT3 and perforin decreased simultaneously with cytotoxicity and dose dependently when sHLA-G1 (1.6 micro g/mL-1.6 ng/mL) was added to IL-2 stimulated cultures. sHLA-G1 did not induce apoptosis and CD25 expression was not affected. CONCLUSION: Interleukin-2R alpha expression is not controlled by sHLA-G1, but its signal transducer STAT3 as well as several downstream effects, such as perforin expression, proliferation and cytotoxicity. The control of STAT3 bioavailability through sHLA-G1 may be a key regulator of the mentioned effects. PMID- 17076672 TI - Association of unexplained infertility with gonadotropin and ovarian antibodies. AB - PROBLEM: To determine the prevalence and characteristics of gonadotropin autoantibodies (GAB) associated with unexplained infertility, and to assess the relationship between ovarian autoantibodies (OVAB) and GAB. METHOD OF STUDY: Ovarian antibodies and GABs in sera of patients with unexplained infertility (n = 53) and a comparison (population) group from a blood bank (n = 40) were detected by immunoassay. Patients with unexplained infertility had either no prior gonadotropin treatment (n = 15) or two or more gonadotropin cycles to induce ovulation (n = 38). RESULTS: The GABs were detected in 67% of women with treatment, 27% of women without treatment and 8% of women in the population. The GABs recognized follicle stimulating hormone (FSH) and luteinizing hormone (LH) and their alpha and beta subunits and to a lesser extent thyroid stimulating hormone (TSH) and prolactin. There was no significant difference in OVAB prevalence between gonadotropin treated or untreated women while GABs were significantly more frequent in gonadotropin treated women (P < or = 0.01). CONCLUSION: Gonadotropin antibodies may represent a separate marker of ovarian autoimmunity in unexplained infertility as they are correlated with OVAB and are present in patients not treated with gonadotropin. However, as a higher frequency of GAB is associated with gonadotropin treatment, patients with ovarian autoimmunity may tend to have an immune response to gonadotropins in addition to an endocrine response. PMID- 17076673 TI - Aberrant uterine natural killer (NK)-cell expression and altered placental and serum levels of the NK-cell promoting cytokine interleukin-12 in pre-eclampsia. AB - PROBLEM: Natural killer (NK) cells are the most abundant lymphocyte population at the maternal-fetal interface. They are suggested to be important during placentation by controlling trophoblast invasion. If placentation is suboptimal, pre-eclampsia can occur. METHOD OF STUDY: Decidual NK (dNK) cells were examined at delivery in 46 women, 22 pre-eclamptic women and 24 healthy controls, by staining for CD56 and CD94 with immunohistochemistry (IHC). Furthermore, we investigated the placental expression and the serum levels of the NK-cell activating cytokines interleukin(IL)-12, IL-15, IL-18 and the anti-inflammatory cytokine IL-10 by IHC and enzyme-linked immunosorbent assay (ELISA), respectively. RESULTS: Pre-eclamptic women had higher number of CD56+ and CD94+ cells in the decidua, indicating an altered receptor expression of dNK cells. We also demonstrate for the first time that the villous trophoblasts show strong immunostaining for IL-12 in placentae from healthy controls, while women suffering from pre-eclampsia have significantly less IL-12. However, pre eclamptic women had significantly elevated IL-12 and IL-15 levels in serum. CONCLUSION: Results show increased numbers and altered phenotype of dNK cells in pre-eclampsia, supporting the importance of these cells for a healthy pregnancy. The altered receptor expression of dNK cells together with diminished placental IL-12 expression could implicate an altered NK cell-regulation in pre-eclampsia. PMID- 17076674 TI - Induction of maternal tolerance to fetal alloantigens by RANTES production. AB - PROBLEM: Previous studies have demonstrated a requirement for RANTES (regulated on activated normal T-cell expressed, and secreted) at immune privileged sites; we have investigated the role of RANTES in the induction of maternal-fetal tolerance. METHOD OF STUDY: Endometrial and peripheral T lymphocytes were obtained from women with recurrent pregnancy losses (RPLs) and fertile women. RANTES modulation by progesterone or paternal alloantigens was measured by enzyme linked immunosorbent assay or flow cytometry analysis. RESULTS: Progesterone significantly increased intracellular RANTES expression in CD4+ and CD8+ endometrial T cells. Moreover, alloreactive lymphocytes from RPL patients produced lower RANTES levels when compared with those from fertile women. At the local level, treatment with recombinant RANTES induced a decrease in CCR5 and CXCR4 messenger RNA that correlated with an increase in T-bet expression. RPL patients and normally fertile women express RANTES similarly, but differ in their patterns of RANTES receptor expression. CONCLUSION: RANTES may be implicated in the local induction of a Th1-type response necessary for successful implantation. Altered response to RANTES stimulation among some RPL patients may be responsible for poor pregnancy outcomes. PMID- 17076675 TI - Expression of natural cytotoxicity receptors and a2V-ATPase on peripheral blood NK cell subsets in women with recurrent spontaneous abortions and implantation failures. AB - PROBLEM: Natural cytotoxicity receptors (NCRs) are unique markers, which regulate NK cell cytotoxicity and cytokine production. a2V-ATPase is expressed on subsets of PBMC and regulates the extracellular environment, which facilitates NK cytotoxicity or cytokine secretion. In this study, we aim to investigate the expression of NCRs and a2V-ATPase in peripheral blood NK cells of women with recurrent spontaneous abortions (RSA) or implantation failures. METHOD OF STUDY: Peripheral blood NK cells (CD56(dim) and CD56(bright) were analyzed for the expression of NCRs (NKp46, NKp44 and NKp30) and a2V-ATPase using 3-color flow cytometry in women with RSA (n=24), implantation failures (n=19) or normal healthy women (n=13). RESULTS: CD56+/NKp46+ cells were markedly decreased (P<0.05) and CD56(bright)/a2V-ATPase+ cells were significantly increased (P<0.05) in women with RSA as compared to those of normal controls. In women with RSA or implantation failures, expression of NKp46, NKp44, NKp30, and a2V-ATPase on CD56(bright) NK cells was significantly up-regulated as compared with those of CD56(dim) NK cells. CONCLUSION: The differential expression of NCRs and a2V ATPase in NK cell subsets may suggest dysregulation of NK cytotoxicity and cytokine production in women with RSA and implantation failures. PMID- 17076676 TI - Endogenous transforming growth factor-beta inhibits toll-like receptor mediated activation of human uterine natural killer cells. AB - PROBLEM: Toll-like receptors (TLRs) recognition is an important means for the innate immune system to rapidly respond to pathogen invasion. Our aim was to determine whether uterine natural killer (uNK) cell cytokine production induced by stimulation through TLRs could be regulated by endogenous transforming growth factor (TGF)-beta in human endometrium. METHOD OF STUDY: Single cells were isolated from human endometrium, and interferon (IFN)-gamma production by endometrium cells and uNK cells was determined after stimulation by TLR agonists. The role of TGF-beta in regulating this response was tested by blocking TGF-beta function using antibodies or a specific inhibitor, SB431542. RESULTS: TGF-beta blockade increased TLR agonist induced IFN-gamma by uNK cells. The regulation of uNK cell cytokine production was observed when uNK cells were incubated with agonists for TLR2 (PGN) or TLR3 (polyI:C). Blockade of TGF-beta or TGF-beta receptor signaling had no effect on constitutive cytokine production in the absence of TLR agonists. CONCLUSION: The results indicate that endogenous TGF beta alters cytokine responses of uNK cells in human endometrium in response to TLR agonists. These data suggest that uNK cell responses to microbial pathogens in the endometrium are regulated by the amount of biologically active TGF-beta present within the human endometrium. PMID- 17076677 TI - B-cell responses to lipopolysaccharide epitopes: Who sees what first - does it matter? AB - PROBLEM: Cholera is the paradigm for waterborne bacterial diseases. For over a 100 years, scientists have tried to develop a universally effective vaccine for cholera. We are hampered in our efforts because we do not know the details of the basic immune response to Vibrio cholerae antigens. What are the most proactive antigens? What special needs for immunization are engendered by previous exposure to cholera or the age of the individual? How long does immunity last, and is this immunity a classic immunologic memory or re-exposure and continual boosting? METHOD OF STUDY: Immunization with synthetic derivatives of the carbohydrate moieties of V. cholerae lipopolysaccharide (LPS) coupled to different carrier proteins (neoglycoconjugates, NGC) has allowed dissection of the response to the disaccharide array of perosamine that represent either the Inaba or the Ogawa serotype. Studying serum anti-LPS endpoint titers and the serum vibriocidal response to NGC provides insight into the importance of LPS serotype-specific B cell epitopes and how antibody response are influenced by the form of the LPS immunogen. RESULTS: We found that murine serum antibody responses to V. cholerae LPS are dynamic. The magnitude of serum anti-LPS antibody titers and the capacity to induced vibriocidal antibodies (immunoglobulin M) are influenced by the initial immunizing serotype of LPS, the structure of the LPS immunogen (native LPS versus NGC), and the order of serotype immunization in a prime boost immunization strategy. The dynamic of the immune response to LPS immunogens is typified by the fact that the host species can affect the immunization response. We found mice do not make vibriocidal antibody to Inaba NGC but rabbits do. This is in contrast to the Ogawa NGC that induced vibriocidal antibody in mice. CONCLUSION: The results suggest that the host's B-cell repertoire can influence the immunization efficacy; therefore, the development of the new generation of NGC V. cholerae vaccines should focus on human volunteers and their ability to mount protective responses. PMID- 17076678 TI - Autoantibodies and prediction of reproductive failure. AB - PROBLEM: To determine which autoantibodies are associated with reproductive failure. METHOD OF STUDY: Sera from 269 patients with autoimmune disease and/or reproductive failure were analyzed for anti-phospholipid (aPL), anti-annexin-V, anti-lactoferrin, anti-thyroglobulin, anti-thyroid peroxidase, anti-prothrombin, anti-nuclear, and anti-saccharomycetes cerevisiae antibodies (ASCA), by enzyme linked immunosorbent assay. Patients were classified as: recurrent pregnancy loss (RPL), infertility, and autoimmune diseases. The results were compared with those of 120 healthy volunteers. RESULTS: In autoimmune diseases, the prevalence of anti-prothrombin, anti-annexin, anti-phospholipid and anti-nuclear antibodies was significantly higher than in the control group, OR 11.0 [CI, 3.5-35.2], 33 [CI, 7.2-174.2], 13 [CI, 1.4-309.7], and 16.1 [CI 2.4-122], respectively. In infertility, the antibodies with significantly higher levels than controls were: aPL OR, 5.11 [CI 1.2-25.4], and anti-prothrombin antibodies, OR, 5.15 [CI, 2.1 12.7]. In RPL, ASCA, anti-prothrombin and aPL were more prevalent than in controls, OR 3.9 [CI, 1.5-10.6], 5.4 [CI, 2.4-12.5] and 4.8[CI, 1.2-22.2] for each antibody, respectively. Anti-prothrombin antibodies and aPL were more significantly associated with late pregnancy losses than early losses. CONCLUSION: ASCA antibodies have not previously been described in RPL. Nor are anti-prothrombin antibodies usually assessed in infertility or RPL. If these results are confirmed in further studies, these antibodies might be assessed routinely in reproductive failure. PMID- 17076679 TI - Specific isolation of placenta-derived exosomes from the circulation of pregnant women and their immunoregulatory consequences. AB - PROBLEM: One immunoregulatory pathway receiving little attention is placental exosome release. In normal pregnancy, as factors linked with early immunomodulation decline, placental exosomes become critical in modulating T-cell activation, suppressing effector T cells by enhancing lymphocyte apoptosis and CD3-zeta loss. METHOD OF STUDY: Placental exosomes were specifically isolated from the maternal peripheral circulation by a chromatographic/immunosorbent procedure. Exosomal suppression of T-cell signaling molecules on unfractionated T cells and T subsets was analyzed by Western immunoblot. The role of Fas ligand (FasL) was defined by use of Fas-blocking antibody. RESULTS: While exosomes of lymphoid origin could be demonstrated in all women, placenta-derived exosomes were only identified in pregnant patients. Placental exosomes suppressed T-cell expression of CD3-zeta and JAK3, while inducing SOCS-2. This downregulation of CD3-zeta was partially reversed by pre-incubating T cells with ZB4 antibody. Using T subsets, the level of CD3-zeta on CD8+ cells was inhibited 1.43-fold more than in CD4+ cells. On CD4+ CD25+ cells, CD3-zeta was not significantly inhibited. CONCLUSION: Placental exosomes suppressed T-cell signaling components; however, while exosomal FasL is an important contributor, it does not appear to be the sole mediator. The additional expression of PD-L1 may explain immunoregulatory consequences of exosomes with low or absent FasL. PMID- 17076680 TI - Impact of microbicides and sexually transmitted infections on mucosal immunity in the female genital tract. AB - Human immunodeficiency virus, genital herpes, and other sexually transmitted infections are a critical national and global health priority requiring the rapid development of safe and effective control methods. Topical microbicides, self administered agents designed for vaginal use, that block transmission at the mucosal surface may provide a realistic method of intervention that could be distributed worldwide. An optimal microbicide should protect against infection but must also be safe, without adversely affecting the mucosal environment, including mediators of host defense. Thus, a critical component in microbicides development is to identify optimal assays that could serve as surrogate markers to predict safety of microbicides prior to embarking on large-scale clinical trials. This will require a greater understanding of the mediators of mucosal immunity in the female genital tract. PMID- 17076681 TI - Controlled ovarian hyperstimulation changes the prevalence of serum autoantibodies in in vitro fertilization patients. AB - PROBLEM: Autoimmune mechanisms are involved in etiology of female infertility, the medical problem frequently treated by in vitro fertilization (IVF). Controlled ovarian hyperstimulation (COH) with supraphysiological levels of sex hormones is achieved by IVF. METHODS OF STUDY: Anti-human-ovary and eight common autoantibodies [nuclear (ANA-H, ANA-R on human HEp-2 cell line and rodent antigen, respectively), smooth muscle (SMA), parietal cell, thyroid microsomal, mitochondrial, beta2-glycoprotein-I, cardiolipin antibodies] found in IVF patients (n = 129) were analyzed with regard to the number of previous IVF procedures and the age of the patient. The changes in autoimmune reactions caused by the COH were determined. RESULTS: Endometriosis and polycystic ovary syndrome were associated with a higher number of common serum autoantibodies compared with the tubal factor infertility (Proportion test, P < 0.05). ANA-R was associated with unexplained infertility [adjusted odds ratio (aOR) 8.79, P = 0.038]. SMA correlated with endometriosis (aOR 37.29, P = 0.008), male factor infertility (aOR 20.45, P = 0.018) and with the previous IVF procedures (aOR 2.87, P = 0.013). There was an overall decrease in the number of detectible autoantibodies after COH (Proportion test, P < 0.05). CONCLUSION: COH may have a suppressive effect on the humoral immunity by the time of embryo transfer but more conclusive studies are needed. PMID- 17076682 TI - Tetrahydrocurcumin: effect on chloroquine-mediated oxidative damage in rat kidney. AB - Tetrahydrocurcumin is an antioxidative substance, which is derived from curcumin, the component of turmeric. In the present investigation, the effect of tetrahydrocurcumin and curcumin against chloroquine-induced nephrotoxicity were studied in female wistar rats. Oral administration of tetrahydrocurcumin significantly prevented the occurrence of chloroquine (970 mg/kg body weight) induced renal damage. Upon administration of tetrahydrocurcumin to chloroquine treated rats, the level of lipid peroxidation was significantly decreased while the levels of non-enzymic and enzymic antioxidants were significantly increased in kidney. Oral administration (80 mg/kg body weight) attenuated the chloroquine induced nephrotoxicity by significantly decreased levels of serum urea and creatinine with significant normalization of creatinine clearance. On administration of tetrahydrocurcumin, the depleted renal antioxidant defense system (enzymatic and non-enzymatic antioxidants) was significantly increased in rats treated with chloroquine. These biochemical observations were supplemented by histopathological examination of kidney section. These results suggest that administration of chloroquine imposes an oxidative stress to renal tissue and that tetrahydrocurcumin protects the oxidative damage associated with chloroquine. PMID- 17076683 TI - Low doses of tumour necrosis factor alpha and interleukin 1beta diminish hepatic gluconeogenesis from alanine in vivo. AB - Previous reports have attributed a stimulating action on hepatic gluconeogenesis to tumour necrosis factor alpha (TNFalpha) administered to rats at high doses (250 mug/kg). However, in adjuvant-induced arthritic rats, which present TNFalpha and other interleukins in the circulation, hepatic gluconeogenesis is diminished. The same occurs in some types of experimental cancer models as, for example, rats bearing the Walker-256 tumour. The present work represents an attempt of reproducing in rats gluconeogenesis inhibition by interleukins using low instead of high doses of both TNFalpha and interleukin 1beta (IL1beta). TNFalpha and IL1beta at doses of up to 10 mug/kg were given endovenously to rats and, after six hours, gluconeogenesis from alanine and several related parameters were evaluated in the isolated haemoglobin-free perfused rat liver. Livers from rats injected with TNFalpha and IL1beta, either alone or in combination, presented diminished gluconeogenesis. The degrees of inhibition caused by TNFalpha+IL1beta, TNFalpha and IL1beta were, respectively, 48.5, 38.8 and 30.4%. TNFalpha also diminished oxygen uptake. No action on urea and ammonia production was found. Possibly, both TNFalpha and IL1beta contribute to the decreased rates of hepatic gluconeogenesis that were found in rats with arthritis, sepsis and some kinds of cancer, but not to the decreased rates of ureagenesis. PMID- 17076684 TI - The effect of oral consumption of perchlorate, alone and in combination with ethanol, on plasma thyroid hormone and brain catecholamine concentrations in the rat. AB - Literature has reported a controversy concerning the effects of the environmental pollutant perchlorate on pertinent physiological systems. However, no research to date has evaluated the effect of concomitant consumption of perchlorate and an additional environmental contaminant on physiological systems. The present preliminary investigation served to assess the effects of oral consumption of perchlorate, alone and in combination with ethanol, on thyroid hormone and brain catecholamine concentrations in female rats of gestational age. Forty, female Myers' high ethanol-preferring rats were randomly assigned to 1 of 7 groups that received: (1) deionized water, both bottles (2) deionized water and 10% ethanol (v/v), two separate bottles (3) 300 microg/l perchlorate solution in deionized water, both bottles (4) 300 microg/l perchlorate in deionized water and in 10% ethanol (v/v), two separate bottles (5) 3000 microg/l perchlorate solution in deionized water, both bottles (6) 3000 microg/l perchlorate in deionized water and in 10% ethanol (v/v), two separate bottles (7) 0.01% propylthiouracil solution in deionized water, both bottles. At cessation of the treatment period, plasma triiodothyronine (T3) and thyroxine (T4) levels were measured by radioimmunoassay and brain area concentrations of dopamine, 3,4 dihydroxyphenylacetic acid (DOPAC), and norepinephrine were measured by high performance liquid chromatography. Perchlorate consumption, alone and/or in combination with ethanol consumption, failed to produce significant alterations from control values for triiodothyronine, thyroxine, dopamine, DOPAC, or norepinephrine. The data suggest that the no-observed effect level of perchlorate consumption on thyroid hormone and brain catecholamine concentrations is above the 3000 microg/l concentration in the adult female rat. PMID- 17076685 TI - Effects of diesel exhaust on lung inflammation related to bacterial endotoxin in mice. AB - We have previously shown that intratracheal instillation of diesel exhaust particles enhances lung inflammation and lung expression of proinflammatory cytokines and chemokines related to bacterial endotoxin (lipopolysaccharide) in mice. The present study was designed to elucidate the effects of inhalation of diesel exhaust on lung inflammation related to lipopolysaccharide. ICR mice were exposed for 12 hr to clean air or diesel exhaust at a soot concentration of 0.3, 1.0, or 3.0 mg/m(3) after intratracheal challenge with 125 microg/kg of lipopolysaccharide. Lung inflammation and lung expression of proinflammatory chemokines such as macrophage chemoattractant protein-1 and keratinocyte chemoattractant were evaluated 24 hr after intratracheal administration. Diesel exhaust inhalation decreased lipopolysaccharide-elicited inflammatory cell recruitment into the bronchoalveolar lavage fluid as compared with clean air inhalation. Histological study demonstrated that exposure to diesel exhaust did not affect lipopolysaccharide-enhanced neutrophil recruitment into the lung parenchyma. Lipopolysaccharide instillation elevated lung expression of macrophage chemoattractant protein-1 and keratinocyte chemoattractant under clean air or diesel exhaust inhalation. However, diesel exhaust exposure did not influence but rather did suppress these levels in the presence of lipopolysaccharide. These results suggest that short-term exposure to diesel exhaust did not exacerbate lung inflammation related to bacterial endotoxin. PMID- 17076686 TI - Protective effect of lignophenol derivative from beech (Fagus crenata Blume) on copper- and zinc-mediated cell death in PC12 cells. AB - Lignophenol, prepared using a phase-separation system, is a derivative of lignin, which is one of the components in the plant cell wall, and possesses high phenolic function, high stability and antioxidant properties. However, little is known about the beneficial effect of lignophenol. In this study, we investigated the protective effect of lignophenol from the beech tree (Fagus crenata Blume) on copper- and zinc-mediated apoptosis in PC12 cells by using DNA fragmentation and TUNEL assays. In DNA fragmentation assays, the DNA ladder patterns in the PC12 cells treated with 200 microM Cu and 200 microM Zn were enhanced, whereas the DNA ladder pattern was hardly observed in these cells treated with 20 mM lignophenol. In the TUNEL assay, TUNEL signals increased significantly in the untreated PC12 cells exposed to 200 microM Cu compared with the control. In contrast, the degree of apoptosis in the 20 mM lignophenol-treated cells was significantly lower than in the untreated cells, indicating that lignophenol inhibited Cu-induced apoptotic cell death in PC 12 cells. In the 200 microM Zn-exposed group, the degree of apoptosis in the 20 mM lignophenol-treated cells was also low compared with the untreated cells. In conclusion, these results suggest that lignophenol plays a role in protecting against Cu- and Zn-mediated PC12 apoptotic cell death. PMID- 17076687 TI - Massive strontium ferrite ingestion without acute toxicity. AB - Ingestion of strontium ferrite is previously unreported. We document absorption of strontium without acute toxicity. A 22 year-old schizophrenic man was brought to hospital after he was witnessed to pulverize and ingest flexible adhesive magnets, which later were identified as strontium ferrite. Other than auditory hallucinations his vital signs, physical examination, ECG and routine laboratories were unremarkable. Abdominal radiographs revealed diffuse radiopaque material. He was treated with whole bowel irrigation with polyethylene glycol electrolyte lavage solution (PEG-ELS) until radiographically cleared. His initial blood and urine strontium levels were 2900 microg/l and 15,000 microg/l, respectively (reference range for urine: <240 microg/l, occupational threshold 800 microg/l). A repeat urine level one week later was 370 microg/l. His hospital course was complicated by bacteraemia secondary to a thrombophlebitis at the site of the intravenous catheter, and the patient was treated with intravenous and oral antibiotics. He remained otherwise asymptomatic and was discharged to a psychiatric unit approximately 3 weeks later. Although clearly absorbed, strontium ferrite does not appear to produce acute toxicity. Delayed, and or chronic toxicity cannot be excluded based on this report. PMID- 17076688 TI - Potassium channels-mediated vasorelaxation of rat aorta induced by resveratrol. AB - Resveratrol, a phenolic substance present in grapes and a variety of medical plants, has been reported to induce vasorelaxation, however the mechanisms are uncertain. In this paper we investigate the possible participation of K(+) channels in the endothelium-independent vasodilatation of rat aorta induced by resveratrol. Resveratrol induced concentration-dependent relaxation of rings with endothelium and without endothelium. We used different potassium channel inhibitors to determine whether the K(+) channels mediated endothelium independent relaxation of rat aorta induced by resveratrol. Highly selective blocker of ATP-sensitive K(+) channels, glibenclamide, as well as non-selective blockers of K(+) channels, tetraethylammonium, did not block resveratrol-induced relaxation of rat aortic rings. Charybdotoxin, a blocker of calcium-sensitive K(+) channels did not affect the resveratrol-induced relaxation. 4-Aminopiridine, non-selective blocker of voltage-gated K(+) (Kv) channels, and margatoxin that inhibits Kv1 channels abolished relaxation of rat aortic rings induced by resveratrol. In conclusion, we have shown that resveratrol potently relaxed rat aortic rings with denuded endothelium. It seems that 4-aminopiridine and margatoxin-sensitive K(+) channels located in the smooth muscle of rat aorta mediated this relaxation. PMID- 17076689 TI - Antispasmodic effects of Rooibos tea (Aspalathus linearis) is mediated predominantly through K+ -channel activation. AB - Rooibos tea has been widely used for abdominal spasm and diarrhoea. The aim of the present study was to explore the possible mechanism for its use in such ailments. Its aqueous extract (RT) at 0.3-10 mg/ml produced relaxation of spontaneous and low K(+) (25 mM)-induced contractions of rabbit jejunum, with weak effect on high K(+) (80 mM)-induced contractions. In the presence of glibenclamide, relaxation of low K(+)-induced contractions was prevented. Cromakalim inhibited contractions induced by low K(+), but not high K(+), while verapamil did not differentiate in its inhibitory effect on contractions produced by the two concentrations of K(+). RT also exhibited antidiarrhoeal and antisecretory activities in mice. The spasmolytic effect was concentrated in organic fractions. Its constituents, chrysoeriol, orientin and vitexin showed a similar pattern of spasmolytic effects to the extract, while rutin was more like verapamil. So Rooibos tea possesses a combination of dominant K(ATP) channel activation and weak Ca(++) antagonist mechanisms and hence justifies its use in hyperactive gastrointestinal disorders. PMID- 17076690 TI - Neuropharmacological analysis of caffeic acid in rats. AB - The aim of the present study was to investigate the effect of intraperitoneal administration of caffeic acid (0.5, 1, 2, 4 or 8 mg/kg) on elevated plus-maze and open field tasks in rats and its possible neuroprotection/neurotoxicity using the comet assay. Caffeic acid at 1 mg/kg increased the number of entries and the time spent in the open arms on plus-maze, suggesting an anxiolytic-like effect when used in lower doses without affecting locomotion and exploration on the open field. Furthermore, a protective effect against hydrogen peroxide-induced oxidative damage on brain tissue was observed through the treatment with caffeic acid at 1 and 8 mg/kg. However, in the highest dose, caffeic acid induced DNA damage in brain tissue. PMID- 17076691 TI - Effect of the food flavour cinnamaldehyde on the antioxidant status of rat kidney. AB - Cinnamaldehyde, a food flavour, has a high potential for human consumption in India. In this study, we evaluated the effect of cinnamaldehyde on the antioxidant status of the rat kidney. Rats were given cinnamaldehyde orally by gavage at dose levels of 2.14, 6.96, 22.62 and 73.5 mg/kg body weight/day for the period of 10, 30 and 90 days. The non-enzymatic antioxidants ascorbic acid, alpha tocopherol and reduced glutathione were decreased while the antioxidant enzymes, superoxide dismutase, glutathione peroxidase and glutathione-s-transferase were increased. Catalase was decreased and thiobarbituric acid-reactive substances were increased only in the kidney of rats treated with cinnamaldehyde at the dose level of 73.5 mg/kg body weight/day during an exposure period of 90 days and not in the kidney of other cinnamaldehyde-treated rat groups. Thus, cinnamaldehyde has an effect on the antioxidant status of rat kidney and its effect is time- and dose-dependent. PMID- 17076692 TI - The effect of combined administration of ethanol and gabapentin on rabbit electroencephalographic activity. AB - The central effect of ethanol is mainly connected with the effect on GABAergic, glutamatergic, serotonergic and opioid transmission. The mechanism of gabapentin effect suggests that it may alleviate the rewarding effect of ethanol, which may be used in the treatment of addiction. We decided to examine the interaction of ethanol with gabapentin by a pharmaco-electroencephalographic (EEG) method. The influence of gabapentin on the effect of ethanol on EEG of rabbits (midbrain reticular formation, hippocampus, frontal cortex) was tested. Gabapentin was administered at a single dose (25 and 100 mg/kg orally) or repeatedly twice a day at a total dose of 25 mg/kg for 14 days. Ethanol was injected at a dose of 0.8 g/kg 60 min. after gabapentin treatment. Ethanol caused an increase in the slow frequencies (0.5-4 Hz) in the recording, as well as a marked decrease of the fastest frequencies (13-30 and 30-45 Hz). Gabapentin lead to changes in rabbit EEG recording suggesting an depressant effect on the CNS (increase of slow and decrease of fast frequencies). The effects were less pronounced after repeated doses, which may indicate adaptative changes in the receptors. Gabapentin administered both in a single dose and for 7 days markedly enhanced the effect of ethanol on EEG recordings in rabbits. Repeated doses of gabapentin decrease the sensitivity of the hippocampus to the effect of ethanol. PMID- 17076693 TI - Predictors of refill non-adherence in patients with heart failure. AB - AIM: To identify the health beliefs and patient characteristics associated with medication non-adherence in patients attending a heart failure outpatient clinic. METHODS: A survey was administered to 350 consenting clinic patients. Questions focused on relevant demographic and clinical characteristics, the Health Belief Model, the Beliefs About Medicines Questionnaire and the Multidimensional Health Locus of Control. Multivariate logistic regression was used to identify independent predictors of refill non-adherence (<90%). RESULTS: Refill non adherence was found in 77 (22%) participants. Being a smoker [odds ratio (OR) 2.4, 95% confidence interval (CI) 1.0, 5.8, P = 0.045], two or fewer medication administration times (OR 2.4, 95% CI 1.2, 4.6, P = 0.01), and positive response to 'Have you changed your daily routine to accommodate your heart failure medication schedule' (OR 2.4, 95% CI 1.2, 4.5, P = 0.01) were the independent predictors of refill non-adherence. CONCLUSION: Perceptions regarding barriers to medication taking and fewer administration times could result in medication non adherence in congestive heart failure patients. Medication regimens should be designed after accounting for patients' existing routines. PMID- 17076694 TI - Risk factors for medication misadventure among residents in sheltered housing complexes. AB - AIM: To identify risk factors for unplanned hospitalizations among residents of sheltered housing complexes (SHCs). METHODS: Medication-related risk factors for health outcomes among residents of SHCs in Aberdeen (n = 1137) were assessed using a postal questionnaire. Predictors of unplanned hospitalization/emergency department (ED) visit were identified using logistic regression. RESULTS: Of the 695 (61.1%) responses received, 645 were from residents (mean age 78.2 years) using prescribed medications. One or more risk factors for medication-related problems was seen in 467 (72.4%) respondents; 488 (75.7%) were using medications with high potential for adverse drug reactions (ADRs) in the elderly. Unplanned hospitalizations/ED visits (n = 230) were found to be associated with use of drugs of narrow therapeutic index [P < 0.001; odds ratio (OR) 2.98, 95% confidence interval (CI) 1.69, 5.28]; use of five or more different medications (P = 0.001; OR 2.10, 95% CI 1.34, 3.31); and greater disability (Townsend score) (P = 0.005; OR 1.06, 95% CI 1.02, 1.11). CONCLUSION: Residents of SHCs using drugs of narrow therapeutic index, using five or more different medications, and with greater disability warrant periodic monitoring. PMID- 17076695 TI - Pharmacokinetics and safety assessments of high-dose and 4-week treatment with S 3304, a novel matrix metalloproteinase inhibitor, in healthy volunteers. AB - AIMS: To investigate the tolerability, safety and pharmacokinetics of S-3304 in healthy volunteers treated with high doses of S-3304 for 28 days. METHODS: Thirty two healthy volunteers were recruited. Four male and four female subjects were allocated to one of four doses (800 mg, 1600 mg, 2400 mg and 3200 mg). At each dose six volunteers took active medication and two volunteers took placebo in a double-blind fashion. Volunteers took a single dose on days 1 and 28 for pharmacokinetic purposes, and took twice daily doses from day 3-27. The pharmacokinetics of S-3304 and its hydroxy metabolites were evaluated. Tolerance was based on subjective adverse events, clinical examination, vital signs, ECG and laboratory tests including haematology and biochemistry profiles using CTC grading. RESULTS: Doses up to 2400 mg twice daily were generally well tolerated. At 3200 mg twice daily, five volunteers including one randomized to placebo were withdrawn from treatment mainly due to alanine aminotransferase (ALT) elevation. C(max) of S-3304 on day 1, whose geometric mean and 95% confidence interval were 66.3 microg ml(-1) (48.8, 90.0) for 800 mg, 82.6 microg ml(-1) (69.3, 98.6) for 1600 mg, 89.5 microg ml(-1) (79.5, 100.7) for 2400 mg, and 110.5 microg ml(-1) (88.9, 137.7) for 3200 mg, respectively, was correlated with the log-transformed peak ALT (P < 0.0001 for male and P = 0.048 for female volunteers). CONCLUSIONS: In healthy volunteers the maximum tolerated dose of S-3304 was 2400 mg twice daily. ALT elevation was the most frequent dose-limiting factor and was correlated with C(max) on day 1. PMID- 17076696 TI - A multiple dose study of prasugrel (CS-747), a novel thienopyridine P2Y12 inhibitor, compared with clopidogrel in healthy humans. AB - AIMS: This double-blind, placebo-controlled trial was designed to evaluate the pharmacodynamics, pharmacokinetics, safety, and tolerability of prasugrel (CS 747, LY640315), a novel thienopyridine P2Y(12) ADP receptor antagonist compared with clopidogrel, during multiple oral dosing in healthy subjects. METHODS: Thirty subjects received placebo, prasugrel 5 mg, 10 mg, or 20 mg, or clopidogrel 75 mg orally, daily for 10 days. Platelet aggregation, bleeding time, and prasugrel metabolites were measured and adverse events were recorded. RESULTS: Inhibition of ADP-induced platelet aggregation reached steady state by day 3 following prasugrel 10 and 20 mg compared with 5 days for clopidogrel 75 mg or prasugrel 5 mg. Compared with placebo, at 24 h after the last dose of study drug, inhibition of platelet aggregation using (20 microm) ADP was significantly higher in the prasugrel 10 mg group (58.2 +/- 4.9% vs. 9.2 +/- 4.0%, P < 0.001) with no difference in the clopidogrel group (15.7 +/- 6.8% vs. 9.2 +/- 4.0%, P = 0.78). With 5 microm ADP, inhibition of platelet aggregation with prasugrel 10 mg and clopidogrel 75 mg was significantly higher than with placebo (prasugrel 10 mg, 70.5 +/- 4.7%; clopidogrel 75 mg, 36.5 +/- 9.0%; vs. placebo, 11.3 +/- 5.1%; P < 0.0001 and P = 0.02). On day 10 at 4 h postdose, bleeding time was prolonged with prasugrel 10 mg (prasugrel 10 mg, 706 +/- 252 s vs. placebo, 221 +/- 38 s, P = 0.05) but not with clopidogrel (283 +/- 56 s, P = 0.98). There were no clinically significant bleeding events, serious adverse events, or discontinuations of the study drug. CONCLUSIONS: Compared with clopidogrel 75 mg, prasugrel 10 mg and 20 mg daily for 10 days resulted in more rapid, more consistent, and higher levels of platelet inhibition. PMID- 17076697 TI - A new method to estimate quantitatively seminal vesicle and prostate gland contributions to ejaculate. AB - AIMS: We sought to optimize a quantitative noninvasive method to determine the concentration in their glands of origin of biochemical markers of compartments of the male genital tract as the first step towards validation of a novel method for estimation of drug concentrations in these male genital tract compartments. METHODS: Sixty-eight men participated. We compared four collection devices to split ejaculate into fractions. Fractions were assayed for fructose and prostate specific antigen (PSA) as unique markers of the seminal vesicle and prostate, respectively. Seminal vesicle fructose and prostatic PSA were estimated using a linear regression method, based on fructose-PSA axis intercepts, and compared with an older method which solves a simultaneous series of equations. RESULTS: A five-compartment collection device performed best with mean (95% confidence interval) PSA vs. fructose r(2) of 0.84 (0.71, 0.98, P < 0.001). Using resampling simulations, glandular PSA and fructose estimates were highly variable and often implausible when using only two fractions. Using our method, the prostate contributed 37-44% to the whole ejaculate and the seminal vesicle contributed 55 61%. The novel regression method was highly correlated (r(2) > or = 0.98) with older methods. CONCLUSIONS: We developed a noninvasive quantitative method of male genital tract biochemical marker estimation using a five-compartment tray to collect three to five ejaculate fractions. Our novel regression method is quantitative and more fully developed than older methods. This noninvasive method for determining glandular marker concentrations should be useful to provide quantitative estimates of drug concentrations in these glands. PMID- 17076698 TI - Newer antidepressant drug use in East Asian psychiatric treatment settings: REAP (Research on East Asia Psychotropic Prescriptions) Study. AB - AIMS: Antidepressant use in East Asia is poorly documented. We compared patients given newer and older antidepressants to test the hypothesis, suggested in the literature, that use of newer antidepressants is associated with treatment settings rather than specific diagnostic categories. METHODS: We compared rates of use of older (pre1990) vs. newer antidepressants among 1898 patients identified as antidepressant treated at 21 centres in five East Asian countries (China, Japan, Korea, Singapore, Taiwan) in 2003. Demographics, treatment setting and clinical factors associated with preferential use of newer drugs were tested in univariate and multivariate analyses. RESULTS: Newer antidepressants were included in the treatment regimens of 67.5% (N = 1282/1898) of study subjects. Prescription for newer antidepressants was significantly associated with younger age (z = -4.55, d.f. = 1888, P < 0.001), hospitalization [odds ratio (OR) 1.32, 95% confidence interval (CI) 1.07, 1.64, P < 0.01] and treatment within psychiatric hospitals (OR 1.59, 95% CI 1.27, 2.00, P < 0.001). On multivariate analyses, treatment with newer antidepressants was independently associated with younger age (P < 0.001), country (P < 0.001) and treatment within private hospitals (P < 0.001), but not with sex or diagnosis of affective or anxiety disorders (all P > 0.1). CONCLUSION: Demographic factors and treatment settings appear to influence antidepressant choice more than clinical factors such as diagnosis. PMID- 17076699 TI - Design of expression vectors for RNA interference based on miRNAs and RNA splicing. AB - RNA interference (RNAi) mediates sequence-specific post-transcriptional gene silencing in many eukaryotes and is used for reverse genetic studies and therapeutics. RNAi is triggered by double-stranded small interfering RNAs (siRNAs), which can be processed from small hairpin RNAs generated from an expression vector. In some recently described vectors, the siRNAs are expressed from synthetic stem-loop precursors of microRNAs (miRNAs) driven by polymerase II promoters. We have designed new RNAi vectors, designated pSM155 and pSM30, that take into consideration miRNA processing and RNA splicing by placing the miRNA based artificial miRNA expression cassettes inside of synthetic introns. Like the original miRNA vectors, we show that the pSM155 and pSM30 constructs efficiently down-regulate expression of firefly luciferase and an endogenous gene, phospholipase D2. Moreover, the expression of a coexpressed fluorescent marker is substantially improved by this new design. Another improvement of these new vectors is incorporation of two inverted BsmBI sites placed internal to the arms of the new miRNA-based vectors, so oligos used for cloning are shorter and the cost is reduced. These RNAi vectors thus provide new tools for gene suppression. PMID- 17076700 TI - Characterization of a multifunctional protein disulfide oxidoreductase from Sulfolobus solfataricus. AB - A potential role in disulfide bond formation in the intracellular proteins of thermophilic organisms has recently been ascribed to a new family of protein disulfide oxidoreductases (PDOs). We report on the characterization of SsPDO, isolated from the hyperthermophilic archaeon Sulfolobus solfataricus. SsPDO was cloned and expressed in Escherichia coli. We revealed that SsPDO is the substrate of a thioredoxin reductase in S. solfataricus (K(M) 0.3 microm) and not thioredoxins (TrxA1 and TrxA2). SsPDO/S. solfataricus thioredoxin reductase constitute a new thioredoxin system in aerobic thermophilic archaea. While redox (reductase, oxidative and isomerase) activities of SsPDO point to its central role in the biochemistry of cytoplasmic disulfide bonds, chaperone activities also on an endogenous substrate suggest a potential role in the stabilization of intracellular proteins. Northern and western analysis have been performed in order to analyze the response to the oxidative stress. PMID- 17076701 TI - Spectroscopic characterization of a higher plant heme oxygenase isoform-1 from Glycine max (soybean)--coordination structure of the heme complex and catabolism of heme. AB - Heme oxygenase converts heme into biliverdin, CO, and free iron. In plants, as well as in cyanobacteria, heme oxygenase plays a particular role in the biosynthesis of photoreceptive pigments, such as phytochromobilins and phycobilins, supplying biliverdin IX(alpha) as a direct synthetic resource. In this study, a higher plant heme oxygenase, GmHO-1, of Glycine max (soybean), was prepared to evaluate the molecular features of its heme complex, the enzymatic activity, and the mechanism of heme conversion. The similarity in the amino acid sequence between GmHO-1 and heme oxygenases from other biological species is low, and GmHO-1 binds heme with 1 : 1 stoichiometry at His30; this position does not correspond to the proximal histidine of other heme oxygenases in their sequence alignments. The heme bound to GmHO-1, in the ferric high-spin state, exhibits an acid-base transition and is converted to biliverdin IX(alpha) in the presence of NADPH/ferredoxin reductase/ferredoxin, or ascorbate. During the heme conversion, an intermediate with an absorption maximum different from that of typical verdoheme-heme oxygenase or CO-verdoheme-heme oxygenase complexes was observed and was extracted as a bis-imidazole complex; it was identified as verdoheme. A myoglobin mutant, H64L, with high CO affinity trapped CO produced during the heme degradation. Thus, the mechanism of heme degradation by GmHO-1 appears to be similar to that of known heme oxygenases, despite the low sequence homology. The heme conversion by GmHO-1 is as fast as that by SynHO-1 in the presence of NADPH/ferredoxin reductase/ferredoxin, thereby suggesting that the latter is the physiologic electron-donating system. PMID- 17076702 TI - Characterization of wheat puroindoline proteins. AB - Puroindoline proteins were purified from selected UK-grown hexaploid wheats. Their identities were confirmed on the basis of capillary electrophoresis mobilities, relative molecular mass and N-terminal amino acid sequencing. Only one form of puroindoline-a protein was found in those varieties, regardless of endosperm texture. Three allelic forms of puroindoline-b protein were identified. Nucleotide sequencing of cDNA produced by RT-PCR of isolated mRNA indicated that these were the 'wild-type', found in soft wheats, puroindoline-b containing a Gly ->Ser amino acid substitution (position 46) and puroindoline-b containing a Trp- >Arg substitution (position 44). The latter two were found in hard wheats. Microheterogeneity, due to short extensions and/or truncations at the N-terminus and C-terminus, was detected for both puroindoline-a and puroindoline-b. The type of microheterogeneity observed was more consistent for puroindoline-a than for puroindoline-b, and may arise through slightly different post-translational processing pathways. A puroindoline-b allele corresponding to a Leu-->Pro substitution (position 60) was identified from the cDNA sequence of the hard variety Chablis, but no mature puroindoline-b protein was found in this or two other European varieties known to possess this puroindoline-b allele. Wheats possessing the puroindoline-b proteins with point mutations appeared to contain lower amounts of puroindoline protein. Such wheats have a hard endosperm texture, as do wheats from which puroindoline-a or puroindoline-b are absent. Our results suggest that point mutations in puroindoline-b genes may confer hard endosperm texture through accumulation of allelic forms of puroindoline-b proteins with altered functional properties and/or through lower amounts of puroindoline proteins. PMID- 17076703 TI - Specific membrane binding of the carboxypeptidase Y inhibitor I(C), a phosphatidylethanolamine-binding protein family member. AB - I(C), an endogenous cytoplasmic inhibitor of vacuolar carboxypeptidase Y in the yeast Saccharomyces cerevisiae, is classified as a member of the phosphatidylethanolamine-binding protein family. The binding of I(C) to phospholipid membranes was first analyzed using a liposome-binding assay and by surface plasmon resonance measurements, which revealed that the affinity of this inhibitor was not for phosphatidylethanolamine but for anionic phospholipids, such as phosphatidylserine, phosphatidylinositol 3-phosphate, phosphatidylinositol 3,4-bisphosphate, and phosphatidylinositol 3,4,5 trisphosphate, with K(D) values below 100 nm. The liposome-binding assay and surface plasmon resonance analyses of I(C), when complexed with carboxypeptidase Y, and the mutant forms of I(C) further suggest that the N-terminal segment (Met1 His18) in its carboxypeptidase Y-binding sites is involved in the specific and efficient binding to anionic phospholipid membranes. The binding of I(C) to cellular membranes was subsequently analyzed by fluorescence microscopy of yeast cells producing the green fluorescent protein-tagged I(C), suggesting that I(C) is specifically targeted to vacuolar membranes rather than cytoplasmic membranes, during the stationary growth phase. The present findings provide novel insights into the membrane-targeting and biological functions of I(C) and phosphatidylethanolamine-binding proteins. PMID- 17076704 TI - Influence of modulated structural dynamics on the kinetics of alpha-chymotrypsin catalysis. Insights through chemical glycosylation, molecular dynamics and domain motion analysis. AB - Although the chemical nature of the catalytic mechanism of the serine protease alpha-chymotrypsin (alpha-CT) is largely understood, the influence of the enzyme's structural dynamics on its catalysis remains uncertain. Here we investigate whether alpha-CT's structural dynamics directly influence the kinetics of enzyme catalysis. Chemical glycosylation [Sola RJ & Griebenow K (2006) FEBS Lett 580, 1685-1690] was used to generate a series of glycosylated alpha-CT conjugates with reduced structural dynamics, as determined from amide hydrogen/deuterium exchange kinetics (k(HX)). Determination of their catalytic behavior (K(S), k(2), and k(3)) for the hydrolysis of N-succinyl-Ala-Ala-Pro-Phe p-nitroanilide (Suc-Ala-Ala-Pro-Phe-pNA) revealed decreased kinetics for the catalytic steps (k(2) and k(3)) without affecting substrate binding (K(S)) at increasing glycosylation levels. Statistical correlation analysis between the catalytic (DeltaG( not equal)k(i)) and structurally dynamic (DeltaG(HX)) parameters determined revealed that the enzyme acylation and deacylation steps are directly influenced by the changes in protein structural dynamics. Molecular modelling of the alpha-CT glycoconjugates coupled with molecular dynamics simulations and domain motion analysis employing the Gaussian network model revealed structural insights into the relation between the protein's surface glycosylation, the resulting structural dynamic changes, and the influence of these on the enzyme's collective dynamics and catalytic residues. The experimental and theoretical results presented here not only provide fundamental insights concerning the influence of glycosylation on the protein biophysical properties but also support the hypothesis that for alpha-CT the global structural dynamics directly influence the kinetics of enzyme catalysis via mechanochemical coupling between domain motions and active site chemical groups. PMID- 17076705 TI - Characterization of the protective T-cell response generated in CD4-deficient mice by a live attenuated Mycobacterium tuberculosis vaccine. AB - The global epidemic of tuberculosis, fuelled by acquired immune-deficiency syndrome, necessitates the development of a safe and effective vaccine. We have constructed a DeltaRD1DeltapanCD mutant of Mycobacterium tuberculosis (mc(2)6030) that undergoes limited replication and is severely attenuated in immunocompromised mice, yet induces significant protection against tuberculosis in wild-type mice and even in mice that completely lack CD4(+) T cells as a result of targeted disruption of their CD4 genes (CD4(-/-) mice). Ex vivo studies of T cells from mc(2)6030-immunized mice showed that these immune cells responded to protein antigens of M. tuberculosis in a major histocompatibility complex (MHC) class II-restricted manner. Antibody depletion experiments showed that antituberculosis protective responses in the lung were not diminished by removal of CD8(+), T-cell receptor gammadelta (TCR-gammadelta(+)) and NK1.1(+) T cells from vaccinated CD4(-/-) mice before challenge, implying that the observed recall and immune effector functions resulting from vaccination of CD4(-/-) mice with mc(2)6030 were attributable to a population of CD4(-) CD8(-) (double-negative) TCR-alphabeta(+), TCR-gammadelta(-), NK1.1(-) T cells. Transfer of highly enriched double-negative TCR-alphabeta(+) T cells from mc(2)6030-immunized CD4(-/ ) mice into naive CD4(-/-) mice resulted in significant protection against an aerosol tuberculosis challenge. Enriched pulmonary double-negative T cells transcribed significantly more interferon-gamma and interleukin-2 mRNA than double-negative T cells from naive mice after a tuberculous challenge. These results confirmed previous findings on the potential for a subset of MHC class II restricted T cells to develop and function without expression of CD4 and suggest novel vaccination strategies to assist in the control of tuberculosis in human immunodeficiency virus-infected humans who have chronic depletion of their CD4(+) T cells. PMID- 17076706 TI - Predictive value of phenotypic variables for skin cancer: risk assessment beyond skin typing. PMID- 17076707 TI - Clinical features of late-onset Behcet's disease: report of nine cases. AB - BACKGROUND: Behcet's disease (BD) is usually diagnosed between the second and fourth decades. Onset after 50 years of age is extremely rare. We aimed to analyze the clinical features of late-onset patients with BD. METHOD: The study was conducted from 439 patients diagnosed as BD according to criteria of the International Study Group for BD. Clinical features of patients who were asymptomatic or suffered from only recurrent aphthous stomatitis (RAS) until 50 years of age but fulfilled the diagnostic criteria of BD after this age were reviewed. RESULTS: The age-of-onset was more than 50 years in nine patients (1.56%). Two patients developed erythema nodosum, two developed pathergy positivity, one developed papulopustules, pathergy positivity and ocular symptoms, one developed papulopustules and pathergy positivity, one developed ocular symptoms, one developed papulopustules and ocular symptoms, and one developed erythema nodosum and pathergy positivity as well as oral aphthae and genital ulcerations after the age of 50 years. The neurologic system was involved in two patients. Mucocutaneous symptoms of two patients worsened after interruption of therapy. Two patients with neurologic and ocular involvement flared during the follow up. CONCLUSION: Since the course of the disease is regarded to be relatively mild in mature patients, it is noteworthy that systemic manifestations such as ocular and neurologic involvement and acute flares developed after the age of 50 years in the limited number of patients with late onset BD in our series. PMID- 17076708 TI - Marked improvement induced in photoaged skin of hairless mouse by ER36009, a novel RARgamma-specific retinoid, but not by ER35794, an RXR-selective agonist. AB - BACKGROUND: Photoaging (premature skin aging) results largely from repeated exposure of the skin to ultraviolet (UV) radiation from the sun. Topical all trans retinoic acid (RA), the only agent that has been approved for the treatment of photoaging, has been shown to reverse this process. In this study, we evaluated the pharmacologic effects of novel synthetic retinoids, ER36009 and ER35794, on murine wrinkles induced by UVB. ER36009 is a specific agonist of retinoic acid receptor (RAR)gamma, the most abundant RAR subtype in the skin, while ER35794 is a potent retinoid X receptor (RXR)-selective agonist. METHOD: After a 10-week exposure to escalating doses of UVB irradiation, the animals were treated three times per week with ER36009 (0.0001%, 0.00025%, 0.0005%), ER35794 (0.025%, 0.05%, 0.1%), RA (0.05%) or acetone (control) for 3 weeks. RESULTS: ER36009 exerted a dose-dependent wrinkle-effacing effect, and 0.0005% ER36009 treated skin was significantly different from the control. ER36009 also significantly and dose-dependently increased both epidermal thickness and the area of the dermal repair zone defined by newly synthesized collagen. The effect of 0.0005% ER36009 on photodamaged skin was superior to that of 0.05% RA. In contrast, ER35794 was inactive in this model, though this compound exhibited lower local toxicity than other retinoids. CONCLUSIONS: These data indicate that RARgamma, but not RXR, plays an important role in the improvement of the signs of photoaging, and so a specific RARgamma agonist might be superior to an RAR pan agonist for clinical treatment. We conclude that ER36009 is a candidate for a potent anti-skin-aging agent. PMID- 17076709 TI - Antihistone antibodies in linear scleroderma variants. AB - BACKGROUND: Linear scleroderma occurs as two clinically distinct variants: the frontoparietal en coup de sabre type, and the torso-extremity type. Antihistone antibodies (AHAs), which traditionally are markers for drug-induced lupus, may also be linked to linear scleroderma. METHODS: Retrospective review of all patients presenting with linear scleroderma who had AHA titers measured. RESULTS: A total of 35 patients were identified. Twenty patients with linear scleroderma of the torso and/or extremities comprised 14 pediatric patients ( 0; probing depth < or = 3 mm; attachment loss = 0) and three periodontitis sites (gingival index > 0; probing depth > or = 5 mm; attachment loss > or = 3 mm) of each of the 35 patients at baseline visits and 6-month visits by means of paper strips. Gingival crevice fluid levels of MMP 9 and TIMP-1 were determined by sandwich enzyme-linked immunosorbent assays. RESULTS: The mean amounts of MMP-9 and TIMP-1 in the gingivitis and periodontitis sites sites were significantly higher than in the healthy sites (P < 0.0001). The progressing site was defined as a site that had 2 mm or more attachment loss during the 6-month study period. Gingival crevice fluid levels of MMP-9 were significantly correlated with probing depth, attachment loss, TIMP-1, age, smoking pack years, and viral load values at baseline and 6-month visits (0.0001 < P < 0.001). TIMP-1 levels were only correlated with CD4, viral load, attachment loss, and MMP-9 (0.001 < P < 0.01). Repeated measures analysis of 11 active sites vs. 269 inactive sites indicated that MMP-9 and TIMP-1 levels were significantly higher in active sites than in inactive sites (P < 0.0001). These data indicate that sites with high ginigval crevice fluid levels of MMP-9 and TIMP-1 in HIV positive patients are at significantly greater risk for progression of periodontitis. PMID- 17076773 TI - The infrabony defect and its determinants. AB - BACKGROUND AND OBJECTIVE: The purpose of this study was to assess the defect width of infrabony defects in a cross-sectional study and to evaluate whether the defect width is a function of defect depth. MATERIAL AND METHODS: Complete sets of intra-oral radiographs of patients with severe periodontitis, which exhibited at least one infrabony defect, were digitised and evaluated. The following parameters were measured: depth and width of the infrabony defect, defect angle, and width of the interdental spaces. RESULTS: Fifty-one patients (26 women), ranging from 21 to 73 yr of age (48.5 +/- 13.4 yr), contributed a total of 1272 teeth with 135 infrabony defects (10.6%). Seventeen infrabony defects were located at sites without a neighboring tooth. Infrabony defects were statistically more prevalent in the mandible (n = 82) than in the maxilla (p = 0.013), and more prevalent at mesial sites (n = 92) than at distal sites (p < 0.001). At infrabony defects, the width of interdental spaces at the most coronal extension of the alveolar crest could be measured only at sites with neigboring teeth 2.67 +/- 0.78 mm (range: 1.19-5.70 mm). Analysis failed to reveal a statistically significant difference between defect width at sites with (2.64 +/- 0.82 mm) and sites without (2.76 +/- 0.70 mm) a neighboring tooth. Multilevel regression analysis revealed narrow defect angles to be related to deep infrabony defects, whereas width of the interdental space and distal location were related to wide defects. CONCLUSION: Defect angle depended on defect depth and defect width was not different at sites with or without a neighboring tooth. Even in severe periodontitis, infrabony defects are found only at a minority of teeth. PMID- 17076774 TI - Monitoring periodontal disease status in smokers and nonsmokers using a gingival crevicular fluid matrix metalloproteinase-8-specific chair-side test. AB - BACKGROUND AND OBJECTIVE: With current periodontal diagnostic tools it is difficult to identify susceptible individuals or sites at risk. The aim of this study was to evaluate the efficacy of the matrix metalloproteinase (MMP)-8 specific chair-side dip-stick test in longitudinally monitoring the periodontal status of smoking (S) and nonsmoking (NS) patients with chronic periodontitis, using their gingival crevicular fluid (GCF) MMP-8 concentrations. MATERIAL AND METHODS: Clinical parameters, MMP-8 test results and concentrations were monitored in 16 patients after initial treatment and in 15 patients after scaling and root planing (SRP), every other month, over a 12-mo time period. Progressing and stable sites, and sites with exceptionally high MMP-8 concentrations, were analysed in smokers and nonsmokers. RESULTS: SRP reduced the mean GCF MMP-8 levels, test scores, probing depth (PD), attachment loss (AL) and bleeding on probing (BOP). In sites of periodontal disease progression, the distribution of MMP-8 concentrations was broader than in stable sites, indicating a tendency for elevated concentrations in patients with periodontal disease. The mean MMP-8 concentrations in smokers were lower than in nonsmokers, but in smokers' and nonsmokers' sites with progressive disease, MMP-8 concentrations were similar. Sites with exceptionally elevated MMP-8 concentrations were clustered in smokers who also showed a poor response to SRP. In these sites, the MMP-8 concentration did not decrease with SRP and these sites were easily identified by the MMP-8 test. CONCLUSION: Persistently elevated GCF MMP-8 concentrations may indicate sites at risk, as well as patients with poor response to conventional periodontal treatment (e.g. SRP). MMP-8 testing may be useful as an adjunct to traditional periodontal diagnostic methods during the maintenance phase. PMID- 17076775 TI - Levels of platelet activating factor in gingival crevice fluid following periodontal surgical therapy. AB - BACKGROUND AND OBJECTIVE: Elevated levels of platelet activating factor (PAF), a potent inflammatory phospholipid mediator, have been previously detected in gingival tissues and gingival crevice fluid (GCF) in periodontal disease. However, the role of this mediator during wound healing after periodontal surgery remains unclear. The hypothesis, a relationship between PAF levels and periodontal healing, was tested by measuring PAF levels in GCF samples collected from sites that had undergone guided tissue regeneration (GTR) or flap surgery. MATERIAL AND METHODS: Using a split-mouth design, 30 intrabony defects were randomly assigned to treatment with GTR (group 1) or to flap surgery (group 2). GCF was sampled pre-operatively and at 6-, 12- and 24-wk follow-up evaluation visits. PAF levels in GCF were analyzed by high-performance liquid chromatography (HPLC). RESULTS: Both treatment modalities significantly reduced the probing pocket depth and improved the clinical attachment level (p < 0.01). Compared with pre-operative values, the GCF volume and PAF levels were significantly decreased at postoperative weeks 6, 12 and 24 in both groups (p < 0.01). There were also significant differences in GCF volume and PAF levels at all time points up to 24 wks in both groups (p < 0.01). No statistically significant differences were observed in any of the parameters investigated between the two groups (p > 0.05). CONCLUSION: PAF is detectable in GCF by HPLC and showed a continuous decrease at all the time points monitored following periodontal surgical therapy. This suggests that changes in the levels of this mediator in GCF might be useful for monitoring the progress of periodontal repair and regeneration. PMID- 17076776 TI - Parathyroid hormone modifies human periodontal ligament cell proliferation and survival in vitro. AB - BACKGROUND AND OBJECTIVE: Periodontal ligament (PDL) cells show traits that are typical of osteoblasts, such as osteoblastic marker gene expression and the ability to respond to parathyroid hormone (PTH) stimulation in an osteoblast-like manner with respect to differentiation and local factor production. In the present study, we hypothesized that human PDL cells might respond to PTH stimulation with changes in proliferation and cell survival and thereby provide another mechanism by which PTH might affect the reparative potential of PDL cells. We speculated that the maturation state of the cells and the mode of PTH(1 34) administration would have an impact on the cellular response. MATERIAL AND METHODS: PDL cells were challenged with PTH(1-34) intermittently or continuously at different maturation states. Cell number, 5-bromo-2-deoxyuridine (BrdU) incorporation, DNA fragmentation, nitric oxide production and the duration of the PTH(1-34) effect were determined. RESULTS: Intermittent PTH(1-34) treatment of preconfluent cells caused a significant increase in proliferation and DNA fragmentation, whereas in more mature cells, proliferation was less enhanced while apoptosis was more pronounced than in immature cells. Continuous PTH(1-34) exposure did not alter proliferation in any maturation state but increased DNA fragmentation in preconfluent cells. PTH(1-34) prevented etoposide-induced apoptosis after 6 h but no longer after 24 h. Nitric oxide production was unaffected. CONCLUSION: These results indicate that human PDL cells respond to PTH(1-34) with changes in proliferative and apoptotic signaling in a maturation state-dependent manner. Besides changes in local factor production, these findings provide a further possible mechanism to support the idea that PDL cells possess the potential to be involved in the regulation of dental hard tissue repair. PMID- 17076777 TI - Restraint stress enhances alveolar bone loss in an experimental rat model. AB - BACKGROUND AND OBJECTIVE: The purpose of the present study was to examine the effects of restraint stress on periodontal breakdown resulting from Porphyromonas gingivalis-challenged periodontitis in rats. MATERIAL AND METHODS: To examine the influence of restraint stress on periodontal breakdown, rats were orally challenged with the periodontal pathogen P. gingivalis. Twenty male, specific pathogen-free (SPF) 3-wk-old, Sprague-Dawley rats were divided into four groups: group A (controls), group B (exposed to restraint stress for 12 h/d for 22 d), group C (orally challenged with P. gingivalis), and group D (exposed to restraint stress for 12 h/d for 22 d and orally challenged with P. gingivalis). After 22 d, all animals were killed. The distance from the alveolar bone crest to the cemento enamel junction was determined, concentrations of adrenocorticotropic hormone were measured as stress markers, and atrophy of the thymus and spleen were assessed. In addition, the furcation area of the maxillary molars was examined histologically, while gingival cytokine gene expression was assessed by mRNA using reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: In the restrained group, all stress markers were elevated, and the thymus and spleen were atrophied. Combined restraint stress and oral challenge with P. gingivalis resulted in significantly higher bone loss, and osteoclasts were observed. RT-PCR analysis revealed low cytokine gene expression in the restrained groups. CONCLUSION: These results suggest that the presence of restraint stress significantly enhances the progression of P. gingivalis-challenged periodontitis in rats. PMID- 17076778 TI - Effect of two different approaches for root decontamination on new cementum formation following guided tissue regeneration: a histomorphometric study in dogs. AB - BACKGROUND AND OBJECTIVE: The aim of the present study was to evaluate comparatively the effect of two different approaches for root decontamination on new cementum formation following guided tissue regeneration (GTR). MATERIAL AND METHODS: Nine mongrel dogs were used to obtain bilateral chronic class III furcation defects by placing cotton ligatures around both third mandibular premolars. The teeth were randomly assigned to receive one of the following treatments: scaling and root planing, by means of hand and rotatory instruments, in order to remove soft and hard deposits as well as all root cementum (group A); or removal of only soft microbial deposits, by polishing the root surface with rubber cups and polishing paste, aiming for maximum root cementum preservation (group B). Both groups were treated with GTR, with the use of resorbable polyglycolic-lactic acid membranes (RESOLUT XT). RESULTS: Four months later, data analysis showed that a superior length (mm) (3.59 +/- 1.67 and 6.20 +/- 2.26 for groups A and B, respectively; p = 0.004) and a thicker layer (microm) (18.89 +/- 9.47 and 52.29 +/- 22.48 for groups A and B, respectively; p = 0.001) of new cementum was achieved by keeping the root cementum in place during root decontamination (group B). Regardless of the treatment modality, the new cementum was predominantly of a reparative, cellular extrinsic and intrinsic fiber type. CONCLUSION: Within the limits of the present study, it may be concluded that root cementum preservation may affect the new cementum formation following GTR in class III furcation defects, and the treatment modality did not influence the type of newly formed cementum. PMID- 17076779 TI - Estrogen and alendronate therapies may prevent the influence of estrogen deficiency on the tooth-supporting alveolar bone: a histometric study in rats. AB - BACKGROUND AND OBJECTIVE: The aim of this study was to evaluate histometrically the influence of estrogen deficiency, and its therapies, on the quality of the tooth-supporting alveolar bone. MATERIAL AND METHODS: Seventy-three female rats were randomly assigned to one of the following groups: group 1 (n = 15), sham surgery; group 2 (n = 15), bilateral ovariectomy (OVX); group 3 (n = 14), OVX plus calcitonin (16 IU/kg); group 4 (n = 14), OVX plus estrogen (20 microg/kg); and group 5 (n = 15), OVX plus alendronate (5 mg/kg). Eighty days after surgery, the animals were killed and their mandibles were removed and processed for histology. Bone density (BD) in the furcation area of the first mandibular molar (i.e. the percentage of demineralized bone tissue in a 1,000 microm zone under the furcation) was histometrically obtained. RESULTS: Data analysis demonstrated that estrogen deficiency negatively affected the tooth-supporting bone density (79.45% +/- 4.22 and 55.23% +/- 6.45, for groups 1 and 2, respectively), and that estradiol and alendronate therapies prevented this effect (61.67% +/- 6.87, 78.09% +/- 3.12 and 81.47% +/- 4.58, for groups 3, 4 and 5, respectively). CONCLUSION: Within the limits of this study, it can be concluded that the density of tooth-supporting bone is affected by estrogen deficiency, and that estradiol and alendronate therapies, but not calcitonin, provide protection against this effect. PMID- 17076780 TI - Location of putative stem cells in human periodontal ligament. AB - BACKGROUND AND OBJECTIVE: The origin of cells in the mature periodontium, and the location of their progenitors, are still unknown. It is also unknown whether inflammation influences the number and distribution of these cells within the periodontium. Molecules such as STRO-1, CD146 and CD44 have been identified on a variety of mesenchymal stem cells. The aim of this study was to identify and localize putative stem cells in diseased and healthy human periodontal ligament using cell-surface markers for mesenchymal stem cells. MATERIAL AND METHODS: Healthy and periodontitis-affected teeth were collected, fixed in 10% neutral buffered formalin, decalcified and embedded in paraffin in preparation for immunohistochemistry. Antibodies against STRO-1, CD146 and CD44 were used to identify putative stem cells in the periodontal ligament. RESULTS: Putative stem cells were identified in both healthy and diseased periodontal ligament. They were mainly located in the paravascular region and small clusters of cells were also found in the extravascular region. Wider distributions of these cells were detected in sections of diseased ligament. CONCLUSION: Within the periodontal ligament of both healthy and diseased teeth, cells have been identified consistent with their identification as putative stem cells. The presence of an inflammatory reaction associated with periodontitis may enhance the number of these cells. PMID- 17076781 TI - Cytokine production in human periodontal ligament cells stimulated with Porphyromonas gingivalis. AB - BACKGROUND AND OBJECTIVE: Although some functions and characterizations of human periodontal ligament (hPDL) cells have been reported, the role of hPDL cells in periodontal disease is poorly understood. We have previously reported that hPDL cells produce many kinds of inflammatory cytokines by stimulation with Prevotella intermedia. In this study, we examined the production of cytokines in hPDL cells stimulated with Porphyromonas gingivalis as compared with P. intermedia. MATERIAL AND METHODS: hPDL cells cultured in Dulbecco's modified Eagles's medium (DMEM) containing 10% fetal bovine serum (FBS) and antibiotics. After three to four passages, hPDL cells were stimulated with P. intermedia (ATCC25601) or P. gingivalis (ATCC33277) for 24 h. Total RNA was extracted by ISOGEN and the expression of cytokine mRNA was determined using reverse transcription-polymerase chain reaction. Cytokines in the culture supernatants were assessed by enzyme linked immunosorbent assay. RESULTS: The expression of interleukin-1beta, interleukin-6, interleukin-8, tumor necrosis factor-alpha (TNF-alpha), receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) mRNA was detected in hPDL cells after stimulation with P. gingivalis as well as P. intermedia. There were no significant differences in the kind of cytokines expressed in hPDL cells between P. gingivalis and P. intermedia. However, P. gingivalis induced a significantly higher production of cytokines in hPDL cells than P. intermedia (p < 0.05). CONCLUSION: This study demonstrated that hPDL cells produce many kinds of cytokines as a result of bacterial stimulation, including stimulation with P. gingivalis and P. intermedia. These results suggest that hPDL cells may play a role in cytokine production in periodontal disease. PMID- 17076782 TI - Effects of smoking and drinking habits on the incidence of periodontal disease and tooth loss among Japanese males: a 4-yr longitudinal study. AB - OBJECTIVE: We investigated the risk of periodontal disease and tooth loss, associated with habits of smoking and alcohol consumption, in a longitudinal study. SUBJECTS AND METHODS: The subjects were 1332 Japanese males, 30-59 yr of age, who were free from periodontal disease at the baseline check-up, and who underwent a second check-up 4 yr later. Periodontal disease was diagnosed using the community periodontal index score, based on the clinical probing of pocket depth (> or = 4 mm). Smoking and alcohol consumption patterns were evaluated using a self-administered questionnaire. RESULTS: A dose-response relationship was observed between the amount of smoking and the incidence of periodontal disease in each age group. The overall odds ratios (95% confidence intervals), adjusted for age and alcohol, were 1.51 (0.95-2.22), 1.58 (1.13-2.22) and 2.81 (1.96-4.03), among smokers consuming 1-19, 20 or 21 or more cigarettes per day, respectively, with a significant linear trend (p < 0.0001). A similar association was found between smoking and tooth loss, except for the 50-59-yr-old age group. The adjusted odds ratios were 1.26 (0.60-2.64), 2.01 (1.21-2.32) and 2.06 (1.23 3.48), respectively. A significant linear trend between smoking and tooth loss was also observed (p = 0.01). Ex-smokers showed no significant difference compared with nonsmokers. We also found a significant linear trend between alcohol consumption and tooth loss among 30-39-yr-old subjects, while no relationship was observed between alcohol consumption and periodontal disease. CONCLUSION: Cigarette smoking was found to be an independent risk factor for periodontal disease and tooth loss. Alcohol consumption was a limited risk factor for tooth loss in the younger age group, but was unrelated to periodontal disease. To prevent periodontal disease and tooth loss, health practitioners need to encourage people to stop smoking or not to start. PMID- 17076783 TI - In vivo beta-defensin gene expression in rat gingival epithelium in response to Actinobacillus actinomycetemcomitans infection. AB - BACKGROUND AND OBJECTIVE: Human beta-defensins have been identified in the oral cavity and are predicted to play a role in the defense against pathogenic bacteria. Homologous rat beta-defensins (RBDs) have been identified, but their expression in the oral cavity has not been examined. Therefore, the aim of this study was to investigate the expression of innate immune mediators in the rat gingival epithelium. MATERIAL AND METHODS: Rats were pretreated with antibiotics to depress the normal oral flora, followed by the introduction of Actinobacillus actinomycetemcomitans in their food to allow colonization and the development of periodontal disease. At various time points, animals were killed and the gingival epithelium was extracted. Semiquantitative reverse transcription-polymerase chain reaction was performed to measure RBD and Toll-like receptor (TLR) mRNA levels. RESULTS: Three beta-defensins (RBD-1, -2 and -5) and two TLRs (TLR-3 and -4) are expressed in normal rat gingival epithelium. After the introduction of A. actinomycetemcomitans, RBD-1 and RBD-2 mRNA levels increased for the first week followed by a return to basal levels. No change in TLR mRNA levels was observed. CONCLUSION: The rat model provides a good system for experimental analysis of the innate immune response to periopathogenic bacteria in the oral cavity, as well as the potential role of beta-defensins in the host response to colonization. PMID- 17076784 TI - Relationship between smoking status and periodontal conditions: findings from national databases in Japan. AB - BACKGROUND AND OBJECTIVE: The association between cigarette smoking and periodontitis was examined employing two nationally representative samples of adults in Japan. MATERIAL AND METHODS: Data were derived from the Survey of Dental Diseases (SDD) and the National Nutrition Survey (NNS) in 1999. In the SDD, periodontal conditions were evaluated by calibrated dentists utilizing the Community Periodontal Index (CPI), whereas in the NNS, participants were interviewed on the basis of smoking status by enumerators. Among 6805 records electronically linked via a household identification code, 4828 records of individuals aged 20 yr or older were analyzed. RESULTS: The prevalence of periodontal disease varied significantly by smoking status (p < 0.0001): 39.3%, 49.5% and 47.3% (CPI > or = 3), and 7.9%, 11.7% and 12.4% (a more severe form of periodontitis, CPI = 4), for nonsmokers, former smokers and current smokers, respectively. In adults aged > or = 40 yr (n = 3493), logistic regression models revealed greater probabilities (approximately 1.4 times higher) of periodontitis [CPI > or = 3, odds ratio = 1.38 (1.12-1.71), p = 0.0024] and a more severe form of periodontitis [odds ratio = 1.40 (1.04-1.89), p = 0.0288] in current smokers compared with nonsmokers, following adjustment for possible confounding factors. CONCLUSION: Based on the findings of this study and other numerous reports, cigarette smoking leads to deterioration of periodontal conditions in Japanese adults. PMID- 17076785 TI - Expression of suppressors of cytokine signaling in diseased periodontal tissues: a stop signal for disease progression? AB - BACKGROUND AND OBJECTIVE: Inflammatory cytokines are thought to trigger periodontal tissue destruction. In addition to being regulated by anti inflammatory mediators, their activity is under the control of suppressors of cytokine signaling (SOCS), which down-regulate the signal transduction as part of an inhibitory feedback loop. We therefore investigated the expression of SOCS-1, 2 and -3, and the cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-10, in different forms of human periodontal diseases. MATERIAL AND METHODS: Quantitative polymerase chain reaction (RealTime-PCR) was performed with mRNA from gingival biopsies of control subjects and from that of patients with chronic gingivitis and chronic periodontitis. RESULTS: Our results show that patients with chronic gingivitis and chronic periodontitis exhibit significantly higher SOCS-1, -2 and -3, TNF-alpha and interleukin-10 mRNA expression when compared with healthy controls. The data also demonstrate that SOCS-1 and -3 mRNA expression was higher in tissue from patients with chronic gingivitis than chronic periodontitis, while the levels of SOCS-2, TNF-alpha and interleukin-10 mRNA were similar in these groups. CONCLUSION: The increased expression of SOCS 1, -2 and -3 mRNA in diseased periodontal tissues is believed to be involved in the down-regulation of inflammatory cytokine and Toll-like receptor signaling, and therefore in the attenuation of both the inflammatory reaction and disease severity. Furthermore, it is possible that variation in the levels of SOCS mRNA expressed in different forms of periodontal diseases may determine the stable or progressive nature of the lesions. PMID- 17076786 TI - Diagnostic imaging: from art to science? PMID- 17076787 TI - Association of spinal cord compression seen on magnetic resonance imaging with clinical outcome in 67 dogs with thoracolumbar intervertebral disc extrusion. AB - OBJECTIVES: To determine whether there is an association between the degree of transverse spinal cord compression detected by magnetic resonance imaging following thoracolumbar Hansen type 1 intervertebral disc disease in dogs and their presenting and postsurgical neurological status. METHODS: Medical records of 67 dogs with surgically confirmed Hansen type 1 intervertebral disc disease (2000 to 2004) were reviewed to obtain the rate of onset of disease, duration of clinical signs and presurgical and postsurgical neurological grade. Percentage of spinal cord compression was determined on transverse T2-weighted magnetic resonance images. Linear regression was used to examine the association between spinal cord compression and each of the above variables. Chi-squared tests were used to examine associations among postsurgical outcome and presurgical variables. RESULTS: Eighty-five per cent (57 of 67) of dogs were chondrodystrophoid. Mean spinal cord compression was 53 per cent (sd=219.7, range 14.3 to 84.9 per cent). There was no association between the degree of spinal cord compression and the neurological grade at presentation, rate of onset of disease, duration of clinical signs or postsurgical outcome, with no difference between chondrodystrophoid and non-chondrodystrophoid dogs. CLINICAL SIGNIFICANCE: The degree of spinal cord compression documented with magnetic resonance imaging in dogs with thoracolumbar Hansen type 1 intervertebral disc disease was not associated with the severity of neurological signs and was not a prognostic indicator in this study. PMID- 17076788 TI - Computed tomography study of implantation corridors in canine vertebrae. AB - OBJECTIVES: To define the characteristics of optimum implantation corridors in vertebral bodies C2 to C7 and T10 to S1, using computed tomography examination. METHODS: Measurements were taken from 207 vertebrae from 35 different adult dogs. RESULTS: Implantation corridors of the cervical vertebrae are narrow. The width preserving the transverse hole is less than 2.5 mm in 68.6 per cent of the 86 vertebrae C2 to C6. Dorsal implantation corridors of the last four thoracic vertebrae are narrow, and major anatomical structures are very close to their emergence point. In 63 per cent of the 40 thoracic vertebrae, the right azygos vein is at a distance less than or equal to 1 mm from the vertebral body. The first six lumbar vertebrae have broader corridors. Furthermore, vascular structures are far from the emergence point. The last lumbar vertebra and the sacrum have a broad pedicle, which provides an alternative site for implant placement. CLINICAL SIGNIFICANCE: For the vertebrae L1 to S1, the dorsal implantation can be performed. For the cervical vertebrae, the risk of laceration of the vertebral artery is high. For the last four thoracic vertebrae, the dorsal implantation should not be used. PMID- 17076789 TI - The value of oblique versus ventrodorsal myelographic views for lesion lateralisation in canine thoracolumbar disc disease. AB - OBJECTIVE: To determine the value of oblique versus ventrodorsal myelographic views for lesion lateralisation in canine thoracolumbar disc disease. METHODS: The ventrodorsal and oblique views from 196 lumbar myelograms of dogs with single thoracolumbar disc extrusions or protrusions were blindly and independently reviewed by two of the authors for evidence of lesion lateralisation. Medical records were reviewed for details regarding hemilaminectomy surgery. The side (left versus right) of the surgery and whether or not the disc material was retrieved were noted. RESULTS: Both reviewers lateralised significantly more disc lesions from the oblique views (93 and 95 per cent) than from the ventrodorsal views (59 and 70 per cent) (P<0.001). Using a combination of oblique and ventrodorsal views, 194 (99 per cent) and 195 (99.5 per cent) lesions were lateralised. Unilateral hemilaminectomy was performed in 193 dogs with myelographic lateralisation and in one dog without myelographic lateralisation. The side of spinal cord decompression corresponded with the myelographic findings in all dogs showing lateralisation on myelography. In the dog without myelographic lateralisation, a left (randomly chosen) hemilaminectomy revealed dorsal protrusion of the annulus fibrosus. CLINICAL SIGNIFICANCE: Myelography, including oblique, ventrodorsal and lateral views, is an accurate method for determining lateralisation of extruded or protruded disc material in the vertebral canal before decompressive surgery. Combined oblique and ventrodorsal views are more useful than either view alone and should be routinely obtained in all lumbar myelographic studies when investigating thoracolumbar disc disease. PMID- 17076790 TI - Clinical efficacy and safety of dexmedetomidine and buprenorphine, butorphanol or diazepam for canine hip radiography. AB - OBJECTIVES: To investigate the usefulness of dexmedetomidine for restraint and sedation during hip radiographic examination of hip-extended or stress radiography views when combined with either buprenorphine, butorphanol or diazepam. METHODS: One hundred and twenty-seven client-owned clinically healthy golden retrievers or rottweilers were enrolled in a clinical trial that compared hip-extended and PennHIP methods for diagnosing hip dysplasia and were randomly allocated to receive dexmedetomidine or medetomidine in combination with buprenorphine, butorphanol or diazepam. Subjective assessments were made for response to pain, response to noise, palpebral reflex, muscle tone and overall quality of sedation; non-invasive physiological variables were also recorded. RESULTS: Overall quality of sedation was graded as good or excellent for dogs administered with a combination of butorphanol or diazepam. However, more dogs that received a combination involving buprenorphine had overall a relatively poorer quality of sedation and required additional administration of buprenorphine before the radiographic procedure could commence. Once sedated, clinically sufficient muscle relaxation accompanied by a very low proportion of dogs responding to pain or noise stimuli were observed in all treatment groups. Heart and respiratory rate, and procedure and recovery times were similar for all treatment groups, and no adverse events were observed during the study. CLINICAL SIGNIFICANCE: Dexmedetomidine sedative protocols, particularly in combination with butorphanol and diazepam, can be used effectively and safely in dogs for radiographic procedures. PMID- 17076791 TI - Comparison of the accuracy of two ultrasonographic measurements in predicting the parturition date in the bitch. AB - OBJECTIVES: To compare the accuracy of two ultrasonographic measurements, inner chorionic cavity diameter and biparietal diameter, for the prediction of parturition date in small and medium size bitches. The effects of litter size and fetal sex ratio on the accuracy of the prediction were also investigated. METHODS: Ultrasonographic examinations were performed on pregnant bitches and, depending on the stage of pregnancy, the inner chorionic cavity diameter and biparietal diameter were measured. The mean value of the data of each structure was calculated and the predicted day of parturition was obtained by the application of equations derived from growth curves we previously developed for different size bitches. RESULTS: The parturition term was predicted with similar accuracy during early gestation, by the measurement of inner chorionic cavity diameter, as well as during late gestation, by the measurement of biparietal diameter. Litter size affected only biparietal diameter accuracy, whereas fetal sex ratio did not influence the estimation of the day of parturition. CLINICAL SIGNIFICANCE: Inner chorionic cavity diameter and biparietal diameter were equally reliable parameters in obtaining a reasonably accurate prediction of parturition date at any gestational period at which a bitch is examined. PMID- 17076792 TI - Diagnosis and surgical treatment of a nasopharyngeal cyst in a dog. AB - A 21-month-old boxer dog was presented with clinical signs caused by nasopharyngeal obstruction. A mineralised mass in the nasopharynx was identified by radiography. Computed tomography allowed accurate anatomical localisation of the cystic lesion and identification of an ossified wall. Surgical removal of the mass by a ventral approach resulted in complete resolution of clinical signs. Histopathological examination strongly suggested that the cyst was derived from remnants of Rathke's pouch. PMID- 17076793 TI - Clinical and magnetic resonance imaging features of nasopharyngeal lymphoma in two cats with concurrent intracranial mass. AB - Lymphoma is reported to be the most common nasal and second most common intracranial neoplasm in cats. Intracranial lymphoma may occur as a primary central nervous system lymphoma or as part of multi-centric disease. Two cats were presented with histopathologically confirmed nasopharyngeal lymphoma and concurrent mass within the middle fossa of the cranial cavity, with magnetic resonance imaging suggestive of direct communication. Both cats demonstrated evidence of bilateral oculomotor nerve deficits and upper respiratory tract noise. In one cat, bilateral optic nerve deficits were also present. The magnetic resonance imaging features were similar in both cases and demonstrated a contrast enhancing intracranial mass on the ventral aspect of the middle fossa of the cranial cavity and an adjacent mass arising from the dorsal aspect of the nasopharynx. Lymphoma should be included as an important differential diagnosis in cats presented with middle cranial fossa syndrome (in particular ophthalmoplegia) and stertor. PMID- 17076794 TI - Nasal melanosis in three dogs. AB - Cytologically and histologically confirmed nasal melanosis was detected by rostrocaudal rhinoscopic evaluation of three dogs with unilateral nasal discharge caused by a chronic and severe odontopathic rhinitis. The extraction of affected teeth and prolonged antibiotic therapy led to a complete resolution of nasal disease. The nasal melanosis could be considered a partial metaplastic transformation of mucosal respiratory cells with accumulation of intracytoplasmic melanin. PMID- 17076795 TI - What is your diagnosis? Small intestinal and colonic obstruction; emphysematous pyometra. PMID- 17076797 TI - A model to describe the relationship between knowledge, skill, and judgment in nursing practice. AB - This paper explores the relationship between knowledge, skill, and judgment and proposes a model to describe that relationship. Through illustration of the components and interrelationships within this model one can more clearly understand the nature of nursing work. Drawing on Benner's work on novice to expert, the model shows the interrelationship and the evolution of knowledge, skill, and judgment in a nurse's practice. PMID- 17076798 TI - Sexuality and Alzheimer's disease: can the two go together? AB - The issue of sexual relations in nursing homes between patients with Alzheimer related dementia is in ward practice largely characterized by confusion and ignorance. Staff are seriously conflicted on how to respond and almost totally untrained on the subject. Patients are being mistreated and humiliated. Central to this state of affairs is insufficient awareness of the ethical and human rights elements in nursing care. Two case studies illustrate the practical dilemmas nurses must solve. A decision-making process is set out to reach an ethical and practical conclusion. Recommendations for action by nurses, the nursing profession, nursing home owners, and policy-makers are given. PMID- 17076799 TI - A concept analysis of voluntary active euthanasia. AB - TOPIC: Euthanasia has a wide range of classifications. Confusion exists in the application of specific concepts to various studies. PURPOSE: To analyze the concept of voluntary active euthanasia using Walker and Avant's concept analysis method. SOURCES OF INFORMATION: A comprehensive literature review from various published literature and bibliographies. CONCLUSIONS: Clinical, ethical, and policy differences and similarities of euthanasia need to be debated openly, both within the medical profession and publicly. Awareness of the classifications about euthanasia may help nurses dealing with "end of life issues" properly. PMID- 17076800 TI - Said another way. AB - Nurses are being judged on their level of competence by most of their patients at the first nurse-patient encounter. Unfairly or not, the nurse's effectiveness in delivering care is determined by the way he or she presents to those in their charge. The purpose of the article is to alert nurses to the impact of the introductory phase of that nurse-patient relationship. Body language and voice tone prove to be strong indicators of how that relationship will evolve. The author speaks from the personal perspective of a seasoned nurse and patient. PMID- 17076801 TI - An attack of the plant parasite Cuscuta reflexa induces the expression of attAGP, an attachment protein of the host tomato. AB - Dodder or Cuscutaceae are holoparasitic plants subsisting on other dicotyledonous plants. The infection process is initiated by adherence of Cuscuta prehaustoria to the host surface, followed by penetration attempts by hyphae. In the case of a successful infection, these organs connect the parasite's vascular tissue to that of the host. Here we show that contact of Cuscuta reflexa prehaustoria to tomato induces the expression of a new arabinogalactan protein (AGP), attAGP, in the tomato precisely at the site of dodder attack. We show that attAGP is a plasma membrane-bound cell wall-localized protein. Using the RNAi technique and attAGP targeted virus-induced gene silencing, we observed a correlation between attAGP expression level and force of attachment of the parasite to host tomatoes. If the expression level of attAGP was reduced, the C. reflexa attachment capability was significantly reduced, too. We conclude that C. reflexa infection induced a signal in the host leading to expression of tomato attAGP, which promotes the parasite's adherence. PMID- 17076802 TI - Large-scale histological analysis of leaf mutants using two simple leaf observation methods: identification of novel genetic pathways governing the size and shape of leaves. AB - Observations of cellular organization are essential in understanding the mechanisms underlying leaf morphogenesis. These observations require several preparative steps, such as fixation and clearing of organs, and such procedures are time-consuming and labor-intensive for large-scale analyses. Thus, we have developed simple methods for the observation of leaf epidermal and mesophyll cells. To visualize the epidermis, a gel cast was made of the leaf surface, which was then observed under a light microscope. To visualize the leaf mesophyll cells, leaves were immersed in a solution containing Triton X-100, briefly centrifuged, and then viewed under a light microscope. These methods allowed us to conduct a histological phenome analysis for a large number of known and newly isolated leaf-shape/size mutants of Arabidopsis thaliana by measuring various parameters, including cell number, size, and distribution of cells within a leaf blade. Mutants showed changes in leaf size caused by specific increases or decreases in the number and/or size of cells. In addition, altered cell distributions in the leaf blade were observed, resulting from increases or decreases in the number of cells along the proximo-distal or medio-lateral axis, or recruitment of cells along a particular axis at the expense of other leaf parts. These results provide a phenomic view of the cellular behavior involved in organ size control and leaf-shape patterning. PMID- 17076803 TI - Identification of trans-acting siRNAs in moss and an RNA-dependent RNA polymerase required for their biogenesis. AB - Trans-acting small interfering RNAs (tasiRNAs) are a class of higher-plant endogenous siRNAs that, like miRNAs, direct the cleavage of non-identical transcripts. tasiRNAs derive from non-coding transcripts (TAS) that are converted into dsRNA by a RNA-dependent RNA polymerase (RDR6), following their initial miRNA-guided cleavage. The dsRNA is then processed by a dicer-like enzyme 4 into phased 21-nucleotide siRNAs. To date, tasiRNAs have been identified only in Arabidopsis, and their identity and function in other land plants are unknown. Here, a set of endogenous small RNAs that correspond in a phased manner to a non coding transcript (contig13502) were identified in the moss Pyscomitrella patens. Northern analysis suggests that contig13502-derived small RNAs are expressed in the juvenile gametophyte. In addition, miR390-guided cleavage of contig13502 at two sites flanking the small RNAs cluster was validated by 5' RACE. These cleavages are predicted to provide defined termini for the production of phased siRNAs. To elucidate the biogenesis of identified siRNAs, we cloned and generated knock-out mutants for an RDR6 moss homologue (PpRDR6). These mutants exhibited an accelerated transition from juvenile to mature gametophyte. In addition, RNA blots demonstrated that they lacked contig13502-derived siRNAs, suggesting that PpRDR6 is required for siRNA biogenesis. A target gene, which showed homology to an AP2/EREBP transcription factor, for one phased siRNA, was validated, corroborating its identity as a trans-acting siRNA. Taken together, our data indicate that contig13502 is a novel TAS locus and suggest a role for derived tasiRNAs in the regulation of gene expression in moss. PMID- 17076804 TI - The gibberellin-induced, cysteine-rich protein GIP2 from Petunia hybrida exhibits in planta antioxidant activity. AB - Numerous GAST-like genes have been identified in various plant species. All code for small proteins with a conserved C-terminal region in which 12 cysteines are located in exactly the same positions. We have previously identified five gibberellin (GA)-induced GAST1-like genes in petunia, GIP1-5. GIP2 is expressed in elongating zones, and its suppression in transgenic petunia plants inhibits stem elongation, suggesting a role for the protein in GA-induced cell growth. However, nothing is known about the biochemical activity of GIP2 or any other GAST-like protein. As all contain putative catalytic disulfide bonds (putative redox-active cysteines), we speculated that they might be involved in redox regulation. Expression analysis of GIP2, GIP4 and GIP5 revealed that they are induced by H(2)O(2). To study whether GIP2 modulates H(2)O(2) levels, we generated transgenic petunia plants expressing GIP2 under the regulation of the ubiquitous CaMV 35S promoter. The transgene reduced H(2)O(2) levels in leaves following wounding. It also reduced the levels of H(2)O(2) in guard cells following osmotic stress and ABA treatments, leading to the suppression of stomatal closure. In addition, the transgene promoted stem and corolla elongation. As reactive oxygen species (ROS) are involved in cell elongation, we suggest that GIP2 affects growth by regulating the levels of ROS. As all known GAST-like proteins contain putative redox-active cysteines, they may all act as antioxidants. PMID- 17076805 TI - phyA dominates in transduction of red-light signals to rapidly responding genes at the initiation of Arabidopsis seedling de-etiolation. AB - Contrary to expectations based on the visible phenotypic behavior of seedlings undergoing de-etiolation in response to continuous red light (Rc), previous gene expression profiling showed that one or more of the five-membered phytochrome (phy) family of Arabidopsis, other than phyB, is predominantly responsible for transducing the Rc signals to light-responsive genes. To begin to identify which phys are involved, and to define potential primary targets of phy signaling, we have examined the genome-wide expression profiles of genes responding to Rc within 1 h (early response genes) of initial exposure of dark-grown wild-type, phyA, phyB and phyAphyB double mutant seedlings to the light signal. The data show that phyA has a quantitatively dominant role in Rc-induced expression of these early response genes, that phyB has minimal detectable regulatory activity in the presence of phyA, but assumes a quantitatively larger role in its absence, and that phyA and phyB combined are responsible for the full extent of Rc responsiveness of 96% of these genes. No evidence was obtained of a significant role for the remaining family members, phyC, phyD or phyE, in this process. In striking contrast, Rc-imposed repression of early response gene expression remains quantitatively strong in the phyAphyB double mutant, as well as the monogenic mutants, suggesting a significant role for one or more of the other three phys in this response. Examination of the established or predicted functional roles of the early response genes indicates that genes encoding transcription factors represent the largest single category, at a frequency three times their prevalence genome-wide. This dominance is particularly striking among those genes responding most robustly to the Rc signal, where >50% are classified as involved in transcriptional regulation, suggesting that these may have potentially primary regulatory roles at the interface between phy signaling and the light-responsive transcriptional network. Integration of the present data with those of a previous genome-scale transcriptional analysis of a pif3 mutant, suggests a complex network involving perception and transduction of inductive Rc signals by both phyA and phyB through both PIF3 and other undefined signaling partners to early response genes. PMID- 17076806 TI - Annotation and expression profile analysis of 2073 full-length cDNAs from stress induced maize (Zea mays L.) seedlings. AB - Full-length cDNAs are very important for genome annotation and functional analysis of genes. The number of full-length cDNAs from maize (Zea mays L.) remains limited. Here we report the construction of a full-length enriched cDNA library from osmotically stressed maize seedlings by using the modified CAP trapper method. From this library, 2073 full-length cDNAs were collected and further analyzed by sequencing from both the 5'- and 3'-ends. A total of 1728 (83.4%) sequences did not match known maize mRNA and full-length cDNA sequences in the GenBank database and represent new full-length genes. After alignment of the 2073 full-length cDNAs with 448 maize BAC sequences, it was found that 84 full-length cDNAs could be mapped to the BACs. Of these, 43 genes (51.2%) have been correctly annotated from the BAC clones, 37 genes (44.0%) have been annotated with a different exon-intron structure from our cDNA, and four genes (4.76%) had no annotations in the TIGR database. Expression analysis of 2073 full length maize cDNAs using a cDNA macroarray led to the identification of 79 genes upregulated by stress treatments and 329 downregulated genes. Of the 79 stress inducible genes, 30 genes contain ABRE, DRE, MYB, MYC core sequences or other abiotic-responsive cis-acting elements in their promoters. These results suggest that these cis-acting elements and the corresponding transcription factors take part in plant responses to osmotic stress either cooperatively or independently. Additionally, the data suggest that an ethylene signaling pathway may be involved in the maize response to drought stress. PMID- 17076807 TI - Negative regulation of defense responses in Arabidopsis by two NPR1 paralogs. AB - NPR1 is required for systemic acquired resistance, and there are five NPR1 paralogs in Arabidopsis. Here we report knockout analysis of two of these, NPR3 and NPR4. npr3 single mutants have elevated basal PR-1 expression and the npr3 npr4 double mutant shows even higher expression. The double mutant plants also display enhanced resistance against virulent bacterial and oomycete pathogens. This enhanced disease resistance is partially dependent on NPR1, can be in part complemented by either wild-type NPR3 or NPR4, and is not associated with an elevated level of salicylic acid. NPR3 and NPR4 interact with TGA2, TGA3, TGA5 and TGA6 in yeast two-hybrid assays. Using bimolecular fluorescence complementation analysis, we show that NPR3 interacts with TGA2 in the nucleus of onion epidermal cells and Arabidopsis mesophyll protoplasts. Combined with our previous finding that basal PR-1 levels are also elevated in the tga2 tga5 tga6 triple mutant, we propose that NPR3 and NPR4 negatively regulate PR gene expression and pathogen resistance through their association with TGA2 and its paralogs. PMID- 17076808 TI - Breaking the silence: brain-computer interfaces (BCI) for communication and motor control. AB - Brain-computer interfaces (BCI) allow control of computers or external devices with regulation of brain activity alone. Invasive BCIs, almost exclusively investigated in animal models using implanted electrodes in brain tissue, and noninvasive BCIs using electrophysiological recordings in humans are described. Clinical applications were reserved with few exceptions for the noninvasive approach: communication with the completely paralyzed and locked-in syndrome with slow cortical potentials, sensorimotor rhythm and P300, and restoration of movement and cortical reorganization in high spinal cord lesions and chronic stroke. It was demonstrated that noninvasive EEG-based BCIs allow brain-derived communication in paralyzed and locked-in patients but not in completely locked-in patients. At present no firm conclusion about the clinical utility of BCI for the control of voluntary movement can be made. Invasive multielectrode BCIs in otherwise healthy animals allowed execution of reaching, grasping, and force variations based on spike patterns and extracellular field potentials. The newly developed fMRI-BCIs and NIRS-BCIs, like EEG BCIs, offer promise for the learned regulation of emotional disorders and also disorders of young children. PMID- 17076809 TI - Gamma band activity and its synchronization reflect the dysfunctional emotional processing in alexithymic persons. AB - In the present study, we investigated the gamma band response and its phase synchrony between electrodes in alexithymia, which is characterized by a disability in identifying and describing feelings. Individuals with high and low alexithymia scores were selected according to the scores on the 20-item Toronto Alexithymia Scale. EEG was recorded from alexithymic and nonalexithymic persons viewing emotionally negative or neutral stimuli. Nonalexithymic persons exhibited increased gamma band power and phase synchronization at the 400-450-ms time window when processing emotionally negative stimuli. Neither enhanced gamma band power nor phase synchronization was observed in alexithymic persons in the negative emotion condition. These results suggest that gamma band activity reflects emotional processing, and alexithymic persons may have a deficit in communication between brain regions or in the utilization of memory or emotional information during the processing of emotional stimuli. PMID- 17076810 TI - Attentional capacity for processing concurrent stimuli is larger across sensory modalities than within a modality. AB - One finding in attention research is that visual and auditory attention mechanisms are linked together. Such a link would predict a central, amodal capacity limit in processing visual and auditory stimuli. Here we show that this is not the case. Letter streams were accompanied by asynchronously presented streams of auditory, visual, and audiovisual objects. Either the letter streams or the visual, auditory, or audiovisual parts of the object streams were attended. Attending to various aspects of the objects resulted in modulations of the letter-stream-elicited steady-state evoked potentials (SSVEPs). SSVEPs were larger when auditory objects were attended than when either visual objects alone or when auditory and visual object stimuli were attended together. SSVEP amplitudes were the same in the latter conditions, indicating that attentional capacity between modalities is larger than attentional capacity within one and the same modality. PMID- 17076811 TI - An electrophysiological investigation of indirect semantic priming. AB - In two experiments, direct and indirect semantic priming were measured using event-related potentials. In Experiment 1, participants rated the relatedness between prime and target on a seven-point scale. In Experiment 2, participants simply read the primes and targets as they monitored for a semantic category in probe filler items. Significant direct and indirect N400 priming effects were observed in both experiments. In Experiment 1, the indirect N400 priming effect remained significant when indirectly related and unrelated word pairs were matched for participants' explicit relatedness judgments. In both experiments, the indirect N400 priming effects were preserved when indirectly related and unrelated word pairs were matched on more global and objective measures of semantic similarity. These findings are discussed in the context of current theoretical models of semantic memory and semantic priming. PMID- 17076812 TI - Masked repetition priming and event-related brain potentials: a new approach for tracking the time-course of object perception. AB - This study reports a new approach to studying the time-course of the perceptual processing of objects by combining for the first time the masked repetition priming technique with the recording of event-related potentials (ERPs). In a semantic categorization task ERPs were recorded to repeated and unrelated target pictures of common objects that were immediately preceded by briefly presented pattern masked prime objects. Three sequential ERP effects were found between 100 and 650 ms post-target onset. These effects included an early posterior positivity/anterior negativity (N/P190) that was suggested to reflect early feature processing in visual cortex. This early effect was followed by an anterior negativity (N300) that was suggested to reflect processing of object specific representations and finally by a widely distributed negativity (N400) that was argued to reflect more domain general semantic processing. PMID- 17076813 TI - Repetition priming of possible and impossible objects from ERP and behavioral perspectives. AB - Theorists postulate that two-dimensional objects that can exist in three dimensions (3D, labeled "possible") are represented by a structural description system (SDS). Objects that cannot because of anomalous structural features (labeled "impossible") are not represented by the SDS. To test these assumptions, ERPs and RTs were recorded while subjects classified possible and impossible objects (each presented three times) as left- or right-facing. Repetition of both types of objects led to RT facilitation. ERP repetition priming occurred only for possible objects, which took the form of a long-duration increase in positive amplitude from about 500 ms for second and third relative to first presentations. The data suggest that RT priming of impossible objects depends on part-based structural encoding, whereas ERP priming most likely reflects contact, upon repetition, with a unitized representation that exists only for possible objects. PMID- 17076814 TI - Brain potentials to mathematical syntax problems. AB - Anterior negativities obtained during sentence processing have never been unambiguously reported in the mathematical domain. The reason for this might be that the tasks explored in the mathematical domain have been far from resembling those typically yielding language-related anterior negativities. To test this hypothesis, we explored three mathematical aspects: Order-relevant information, a parenthesis indicating the onset of an embedded calculation, and violations of the type of symbol displayed. Results yielded parieto-occipital instead of frontal negativities. Late posterior positivities were also found, largely comparable to linguistic P600 in topography, but dissociable in functional terms. Our data suggest that language-related anterior negativities may indeed reflect language-specific resources of the human brain and support recent claims that language and mathematical domains are more independent than previously thought. PMID- 17076815 TI - Event-related potentials during mental imagery of animal sounds. AB - To investigate the neural correlates of imagined animal sounds, event-related potentials (ERPs) were recorded while subjects were presented with (1) animal pictures without any imagery instruction (control) or (2) animal pictures with instructions to imagine the corresponding sounds (imagery). The results revealed imagery effects starting with an enhancement of the P2, possibly indexing the top down allocation of attention to the imagery task, and continuing into a more positive-going deflection in the time window of 350-600 ms poststimulus, probably reflecting the formation of auditory imagery. A centro-parietally distributed late positive complex (LPC) was identified in the difference waveform (imagery minus control) and might reflect two subprocesses of imagery formation: sound retrieval from stored information and representation in working memory. PMID- 17076816 TI - Unilateral right-hand contractions cause contralateral alpha power suppression and approach motivational affective experience. AB - Contractions of the left hand induce sadness and bias judgments negatively, whereas contractions of the right hand induce positive affect and assertiveness and bias judgments positively. These results have been explained as resulting from activation of right and left frontal cortices, respectively. However, no research has tested this explanation. The present experiment provided such a test by having participants contract the right or left hand while electroencephalographic activity was recorded. Results indicated that right-hand contractions produced greater left than right frontal activity, whereas left-hand contractions produced greater right than left frontal activity (inverse of alpha power). Similar activations occurred in central regions, perhaps due to mu rhythms. Moreover, as compared to left-hand contractions, right-hand contractions caused greater self-reported approach affect to a mildly positive radio editorial. PMID- 17076817 TI - Brain potentials indicate poor preparation for action in schizophrenia. AB - Impaired antisaccade performance in schizophrenia (SZ) may originate from poor task preparation, suggested by low amplitudes of the contingent negative variation (CNV) before antisaccades. To dissociate components of preparation we measured the CNV in standard pro- and antisaccades and a stimulus preceding negativity (SPN) in delayed pro- and antisaccades. In healthy controls the SPN had lower amplitudes than the CNV, reflecting mere stimulus expectation in SPN and combined stimulus expectation and action readiness in CNV. SZ patients had lower CNV amplitudes than controls, but there was no reliable indication of a general SPN reduction, suggesting a particular impairment of action readiness. The CNV and SPN amplitudes of controls were larger if tasks had incongruent (anti) compared to congruent (pro) S-R mappings. This difference was absent in SZ patients, suggesting a failure to activate specific resources for incongruent S-R mappings. PMID- 17076818 TI - Respiratory sinus arrhythmia, emotion, and emotion regulation during social interaction. AB - Respiratory sinus arrhythmia (RSA) figures prominently in emotional responding, but its exact role remains unclear. The present study tests two hypotheses: (1) Between-person differences in resting RSA are related to emotional reactivity, and (2) within-person changes in RSA are related to regulatory efforts. Pairs of women watched an upsetting film and discussed it. One woman in each of the experimental dyads was asked to either suppress or to reappraise during the conversation. Their partners and both members of the control dyads conversed naturally. Between-person differences in resting RSA were assessed with paced breathing, and within-person changes in RSA were calculated from baseline to the conversation accounting for respiration. Women with higher resting RSA experienced and expressed more negative emotion, and women who attempted to regulate their emotions either by suppressing or reappraising showed larger increases in RSA than controls. PMID- 17076819 TI - Schedule-induced electrodermal responding in children. AB - The current studies were designed to determine whether different intermittent schedules of reinforcement would have a differential effect on the skin conductance responses (SCRs) of children in the 8-12-year-old age range. Results of two experiments indicated that the amplitude of children's SCRs immediately following the occurrence of reinforcement were a function of the schedule of reinforcement, with larger SCRs associated with longer inter-reinforcement intervals. The findings are consistent with the research literature suggesting that long inter-reinforcement intervals can be evocative of aversively motivated emotions, as well as with research on the effects of interstimulus intervals on SCRs. Possible implications for these findings are discussed. PMID- 17076820 TI - The effects of the spontaneous presence of a spouse/partner and others on cardiovascular reactions to an acute psychological challenge. AB - The presence of supportive others has been associated with attenuated cardiovascular reactivity in the laboratory. The effects of the presence of a spouse and others in a more naturalistic setting have received little attention. Blood pressure and heart rate reactions to mental stress were recorded at home in 1028 married/partnered individuals. For 112 participants, their spouse/partner was present; for 78, at least one other person was present. Women tested with a spouse/partner present showed lower magnitude systolic blood pressure and heart rate reactivity than those tested without. Individuals tested with at least one nonspousal other present also displayed attenuated reactivity. This extends the results of laboratory studies and indicates that the spontaneous presence of others is associated with a reduction in cardiovascular reactivity in an everyday environment; spouse/partner presence would appear to be especially effective for women. PMID- 17076821 TI - The impact of emotions on the perception of dyspnea in pediatric asthma. AB - Inadequate symptom perception is increasingly recognized as a contributing factor in asthma morbidity. Recent research has shown that psychological factors can influence the perception of asthmatic symptoms such as dyspnea, but little is known, about the impact of emotions on the perception of dyspnea in pediatric asthma. Therefore, we examined the impact of viewing affective film clips of positive, neutral, and negative valence on perceived dyspnea during resistive load breathing in children with stable mild asthma. Perceived dyspnea decreased during the positive film compared to baseline and neutral film whereas the level of respiratory loading and respiratory resistance (R5) remained unchanged across conditions. The results underline the potential impact of psychological factors such as emotions on the perception of dyspnea in children with asthma. PMID- 17076822 TI - The psychosocial stress-induced increase in salivary alpha-amylase is independent of saliva flow rate. AB - The stress response of salivary alpha-amylase (sAA) has been suggested as an index for sympathetic nervous system activation. However, concurrent inhibition of the parasympathetic nervous system is discussed as a confounder due to suppression of saliva flow rate. Here we set out to test the influence of stress induced changes in flow rate on sAA secretion. Twenty-six subjects underwent the Trier Social Stress Test and a control condition. Saliva was sampled by passive drooling or salivettes. Saliva flow rate, sAA levels and output, salivary cortisol, and heart rate variability were measured. Flow rate increased only when sampled by passive drooling. Stress-induced increases in amylase levels were correlated with increases of amylase output but not with flow rate. Results indicate that flow rate is not a confounder of stress-induced sAA activation and suggest that valid measurements of sAA can be obtained by salivettes without the need for assessment of flow rate. PMID- 17076823 TI - Acute cortisol administration reduces subjective fatigue in healthy women. AB - Treatment with cortisol has been found to decrease fatigue and increase feelings of vigor in both patients and healthy male subjects. We obtained self-reports of mood before 35 mg cortisol or placebo ingestion, 70 min later, and after the healthy female subjects performed cognitive tasks for 1 h in a double-blind within-subject study. Cortisol decreased fatigue, increased vigor, and tended to decrease tension. Effects on fatigue were largest after task performance, when fatigue had increased, suggesting that improvement of fatigue by cortisol is observed when subjects are fatigued. This is the first study to demonstrate improvements in fatigue in healthy female subjects; this is particularly relevant because of the high prevalence of hypocortisolimic fatigue syndromes in women and recent evidence that many psychiatric disorders may involve stress-induced hypocortisolemia that is responsive to cortisol replacement. PMID- 17076824 TI - Sports-related eye injuries: floorball endangers the eyes of young players. AB - The objectives of this study were to determine the distribution of different sports-related eye injuries and to identify injury types to enable recommendations to be made about the use of protective eyewear. The study population comprises all 565 eye trauma patients examined at the Ophthalmology Emergency Clinic of the Helsinki University Central Hospital over a 6-month period. Data were collected from patient histories and questionnaires. In addition, three severe floorball eye injury cases are presented. Of the 565 eye traumas, 94 (17%) were sports related. Of these, 42 (45%) were associated with floorball. Countrywide, in Finland, estimated over 300 (+95% CI 228-415) floorball eye injuries occur annually. The mean age of floorball patients was 22 years. The most common finding (55%) in sports injury patients was hyphema. Clinically severe eye injuries during this period accounted for one-fourth of all cases. During the study period, no eye injury was found in an organized junior ice hockey, where facial protection is mandatory. Floorball is estimated to belong to the highest risk group in sports, and thus, the use of protective eyewear is strongly recommended. We conclude that national floorball federations should make protective eyewear mandatory. PMID- 17076825 TI - Achilles tendon pain intensity and level of neovascularization in athletes as determined by color Doppler ultrasound. AB - The cause of pain in Achilles tendinopathy is thought to be related to the presence of neovascularization in the tendon as seen on color Doppler ultrasound. Asymptomatic pathology has been observed in patellar tendons of elite athletes. The purpose of this study was to examine the prevalence of Achilles tendon pain and the characteristics of Achilles tendons in a young athletic population. Sixty four varsity athletes underwent color Doppler ultrasound imaging to determine tendon thickness, presence of degeneration and neovascularization. The presence of swelling and tenderness was determined, and Achilles tendon pain was rated on three visual analogue scales (VAS) (pain during exercise, pain at rest, pain during daily activities) as well as on the VISA-A scale. Tendon symptoms were not related to the presence of neovascularization. There was a low prevalence of Achilles tendinopathy, tenderness, and neovascularization in this population. Neovascularization was seen in both a painful and a non-painful tendon. PMID- 17076826 TI - Effect of physical activity and muscle morphology on endothelial function and arterial stiffness. AB - Physically active persons have a reduced risk of atherosclerotic disease. Arterial stiffness and endothelial dysfunction are important risk factors for cardiovascular disease. A high proportion of type I (slow-twitch) muscle fibers in skeletal muscle is associated with a favorable cardiovascular risk profile. We tested physical activity and muscle fiber-type distribution as determinants of endothelial function and arterial stiffness in middle-aged men. Fifty-four men (median age 58) who underwent a muscle biopsy in 1984 were re-studied in 2003. Aortic pulse wave velocity (PWV) and pulse wave reflection were assessed by applanation tonometry. Endothelial function was tested by examining the effects of salbutamol and nitroglycerin on pulse wave reflection. In multiple regression analyses aortic PWV (R2=0.51) correlated positively with age (P=0.017), BMI (P=0.001), and systolic blood pressure (P=0.004). A high augmentation index (R2=0.33) was associated with smoking (P<0.001), high LDL cholesterol (P=0.02), and elevated diastolic blood pressure (P=0.03). Impaired endothelial function (R2=0.37) was associated with high age (P=0.04), high LDL cholesterol (P=0.017), high triglycerides (P=0.027), and high physical activity (P=0.005). Muscle fiber type distribution is not a determinant of arterial stiffness or endothelial function. Impaired endothelial function was observed in physically active men, underlining the need for further research. PMID- 17076827 TI - Plasma levels of interleukin-6 and C-reactive protein are associated with physical inactivity independent of obesity. AB - It is recognized that the path from physical inactivity and obesity to lifestyle related diseases involves low-grade inflammation, indicated by elevated plasma levels of inflammatory markers. Interestingly, contracting skeletal muscle is a major source of circulating interleukin-6 (IL-6) in response to acute exercise, but with a markedly lower response in trained subjects. As C-reactive protein (CRP) is induced by IL-6, we hypothesized that basal levels of IL-6 and CRP reflect the degree of regular physical activity when compared with other markers of inflammation associated with lifestyle-related morbidity. Fasting plasma/serum levels of IL-6, IL-18, CRP, tumur necrosis factor-alpha (TNF-alpha), soluble TNF receptor II (sTNF-RII), and adiponectin were measured in healthy non-diabetic men and women (n=84). The amount of leisure-time physical activity (LTPA) was assessed by interview. Obesity was associated with elevated insulin, C-peptide, triglycerides, low-density lipoprotein, IL-6, CRP, and adiponectin (all P<0.05). Importantly, physical inactivity was associated with elevated C-peptide (P=0.036), IL-6 (P=0.014), and CRP (P=0.007) independent of obesity, age, gender, and smoking. Furthermore, the LTPA score was inversely associated with IL-6 (P=0.017) and CRP (P=0.005), but with neither of the other markers. The results indicate that low levels of IL-6 and CRP - not IL-18, TNF-alpha, sTNF-RII, or adiponectin - reflect regular physical activity. PMID- 17076828 TI - Pain and negative mood during rehabilitation after anterior cruciate ligament reconstruction: a daily process analysis. AB - Daily diary methods were used to examine changes in pain and negative mood over the first 6 weeks of rehabilitation after surgical reconstruction of the anterior cruciate ligament (ACL). Participants (58 men and 33 women) completed measures of personal factors (i.e., age, athletic identity, neuroticism, optimism) before surgery and indices of daily pain, negative mood, and stress for 42 days after surgery. Multilevel modeling revealed that, as would be expected, daily pain ratings decreased significantly over the course of the study and that the rate of decline in pain ratings decreased over time. Age and daily negative mood were positively associated with daily pain ratings. Daily negative mood also decreased significantly over the course of the study and was positively associated with neuroticism, daily pain, and daily stress. Athletic identity and optimism interacted with time since surgery in predicting daily negative mood such that participants with high levels of athletic identity and low levels of optimism reported greater decreases in daily negative mood over time. Overall, the findings reveal a pattern of improved psychological functioning over the early stages of post-operative ACL rehabilitation. PMID- 17076829 TI - The double-bundle technique for anterior cruciate ligament reconstruction: a systematic overview. AB - In traditional anterior cruciate ligament reconstruction, there is a subset of patients complaining of knee instability, especially rotational instability, and athletes not able to return to their preinjury level of sports activity. Currently, controversy exists over the usefulness of the double bundle technique (DBT) in addressing these problems. In order to evaluate the DBT, we completed a literature review from 1969 to February 2006 focusing on anatomy, magnetic resonance imaging, graft incorporation, biomechanics, kinematics, surgical techniques, complications and outcome. The DBT is not a standardized technique, which makes it difficult to compare results. Cadaver studies have proven biomechanical advantages with respect to ap-stability, but assessing the rotational stability remains difficult. There is a lack of available outcome studies with sufficient follow-up to demonstrate the potential advantages of DBT. The theoretical advantages of DBT require careful evaluation with outcome, biomechanical and kinematic studies. In addition, studies are needed to address issues such as graft incorporation and complications. An advantage offered by DBT is the possibility to identify rupture patterns that can lead to surgical preservation of an intact and augmentation of an injured bundle. The approach of augmentating a single bundle technique reconstruction with adequate anterior posterior but poor rotational stability is promising. PMID- 17076830 TI - Truncus endurance, hip and ankle mobility and aerobic fitness in 15-year-old Norwegian adolescents in 1968 and 1997. AB - This study compared the physical performance of adolescents in 1997 and 1968. The material included 1329 15-year-old Norwegian adolescents in Akershus county in Norway in 1968, and 1105 15-year-old adolescents in the same region in 1997. The investigations were supervised by the same conductor on both occasions, and used identical physical tests. The test battery included dynamic endurance of abdominal and low back muscles, hip flexion, ankle dorsiflexion and aerobic fitness. Compared with 1968, low back extension endurance in 1997 was almost 10% lower in both genders, the endurance of the abdominal muscles was 4.8% poorer among boys and 8.1% higher among girls and the ratio between abdominal and low back extension endurance was 11.8% higher among boys and 26.4% higher in girls. Compared with 1968, hip flexion in 1997 was 8.3% higher in girls, while ankle dorsiflexion and aerobic fitness in boys were 15% and 13.4% poorer, and 4.8% poorer and 2.5% higher in girls, respectively. The data indicate a general decline during one generation in physical performance among boys, a decline in low back endurance, truncus balance and ankle mobility among girls and in a higher abdominal endurance among girls. PMID- 17076831 TI - Traumatic cervical instability in martial arts. PMID- 17076832 TI - Biological maturity and injury in elite youth football. AB - The purpose of the present study was to investigate injury according to biological maturity in elite under-14 youth football players based at the National Football Institute, France. Over 10 seasons, injury incidence, severity and distribution were compared in 233 players classed according to individual biological maturity determined by skeletal age into three cohorts as early, normal and late maturers.A non-significant higher injury incidence was found in early and normal maturers compared with late maturers. In contrast, the latter group sustained a significantly higher incidence of major injuries compared with early maturers (0.3 vs 0.6 vs 0.9, P=0.039). A significantly higher incidence of osteochrondoses was reported in normal and late maturers (0.3 vs 0.7 vs 0.9, P=0.014), whereas tendinopathy incidence was greater in early and normal maturers (0.06 vs 0.08 vs 0.02, P=0.033). Early maturers incurred the highest incidence of groin strains and re-injuries (P<0.05). There was no significant difference between groups in the seasonal disposition of injury.Biological maturity status did not significantly affect overall injury incidence in elite French youth football players, although there were differences between maturity groups when patterns of injury location, type, severity and re-injury were analyzed. PMID- 17076833 TI - Effect of inversion and ankle bracing on peroneus longus Hoffmann reflex. AB - This study examined peroneus longus (PL) Hoffmann reflex (H-reflex) during sudden inversion perturbation of the ankle/foot complex under an ankle brace and non brace condition. Ten healthy subjects volunteered. H-reflexes were tested on the up-sloping portion of the recruitment curve, utilizing a control trial M-wave above motor threshold to maintain consistency between subjects and conditions. The PL H/maximum M-wave (M(max)) ratio was established using the PL H-reflex and PL M(max) peak-to-peak measures. The mean ratio across five trials for each subject under each ankle brace (brace, no brace) and surface (flat, inversion) conditions was utilized for analysis. The 1 x 4 repeated measures ANOVA revealed a significant main effect for treatment condition (P<0.0001). The PL H/M(max) ratio significantly increased during sudden inversion-no ankle brace condition compared with the flat surface no-ankle brace condition (P=0.04). Application of an ankle brace had no effect on PL H/M(max) ratio during inversion (P=0.78). During this study PL H/M(max) ratios increased during an inversion perturbation in healthy ankles. This is believed to occur due to heightened sensorimotor demand placed on the nervous system during this motion. Moreover, application of an ankle brace during inversion does not appear to affect PL H/M(max) ratio. PMID- 17076834 TI - Passive knee joint range of motion is unrelated to the mechanical properties of the patellar tendon. AB - The physiological factors that govern passive joint range of motion (ROM) are poorly understood. The present study investigated the relation between passive knee joint ROM and the mechanical properties of the patellar tendon. Knee joint ROM was assessed in 43 individuals, and the subjects with the greatest ROM (flexible group, n=10) and lowest ROM (inflexible group, n=10) were selected for further analysis. In these groups an overall "lower extremity joint ROM score" was determined with 11 clinical tests. The elongation of the patellar tendon was assessed during graded maximal isometric knee extensor contractions using ultrasonography, and the mechanical properties of the patellar tendon were determined from corresponding load and tendon deformation data. The two groups were similar with respect to weight, height, tendon cross-sectional area and length, and were, furthermore, equally physically active. The knee joint ROM and lower extremity joint ROM score was significantly different between the groups (flexible: 136+/-7 degrees vs inflexible: 76+/-16 degrees , P<0.001 and flexible: -4.7+/-1.3 vs inflexible: 3.1+/-4.1, P<0.001). There was no difference between groups in maximal knee extensor force or the corresponding tendon deformation. The tendon stiffness (flexible: 3269+/-1591 vs inflexible: 3185+/-1457 N/mm), stress (flexible: 22.4+/-6.5 vs inflexible: 34.0+/-17.6 N/mm(2)), strain (flexible; 6.5+/-1.6 vs inflexible: 7.2+/-1.9%) and Young's modulus (flexible: 0.81+/-0.35 vs inflexible: 1.22+/-0.52 GPa) were not different between the two groups of subjects. These data suggest that differences in knee joint ROM cannot be explained by the mechanical properties of the patellar tendon. PMID- 17076835 TI - Transfusion medicine illustrated. Pica, peanuts, and plasma exchange. PMID- 17076836 TI - Transfusion medicine history illustrated. Transfusion in early medical photography. PMID- 17076837 TI - Microchimerism: a lasting legacy of transfusion? PMID- 17076838 TI - How do I encourage clinicians to transfuse mismatched blood to patients with autoimmune hemolytic anemia in urgent situations? PMID- 17076839 TI - Leukoreduction of blood transfusions does not diminish transfusion-associated microchimerism in trauma patients. AB - BACKGROUND: Transfusion of trauma patients can result in long-term survival of donor white blood cells (WBCs) or "transfusion-associated microchimerism" (TA MC). The aim was to determine whether leukoreduction of blood transfusions, advocated to reduce the immunomodulatory effect of transfusion, decreases the likelihood of developing TA-MC. STUDY DESIGN AND METHODS: A subgroup of trauma patients from a randomized trial was examined, evaluating the risk of infection following leukoreduced versus nonleukoreduced blood transfusion. Patients' blood was sampled at least 1 month after hospital discharge, and TA-MC was assessed with quantitative allele-specific polymerase chain reaction detection of differences at the HLA-DR locus or a panel of insertion-deletion polymorphism loci distributed throughout the chromosomal complement. At the time of blood sampling, a scripted interview was used to ascertain symptoms suggestive of chronic graft-versus host disease (cGVHD). RESULTS: For 67 patients evaluated, the mean age was 43 +/- 17 years and mean Injury Severity Score was 24 +/- 12. Median time from injury to blood sampling for TA-MC was 240 (interquartile range, 116-360) days. Nine of 32 patients (28%) in the nonleukoreduced transfusion group developed TA-MC compared to 13 of 35 patients (37%) in the leukoreduced group (p = 0.43). Subjects with TA-MC were no more likely than subjects without TA-MC to have at least one symptom suggestive of cGVHD (64% vs. 76%, respectively). CONCLUSIONS: TA-MC seems to be a prevalent condition among injured patients at the second of two regional trauma centers evaluated, suggesting that it is a common phenomenon after transfusion in the setting of injury. Although leukoreduction removes greater than 99.9 percent of donor WBCs, it fails to prevent or even substantially reduce the likelihood of developing TA-MC. TA-MC does not appear to be strongly associated with symptoms suggestive of cGVHD several months after transfusion. PMID- 17076840 TI - Enhanced ascertainment of microchimerism with real-time quantitative polymerase chain reaction amplification of insertion-deletion polymorphisms. AB - BACKGROUND: The characterization of microchimerism (MC) by gene amplification has been limited by few allogeneic markers, ascertainment bias, and assay analytic performance. To address this, a panel of 12 MC assays based on insertion-deletion (InDel) polymorphisms had been optimized. STUDY DESIGN AND METHODS: The InDel assays were validated with comprehensive in vitro spiking studies at the stochastic limit of detection. Their ability was also determined to ascertain MC of unknown source genotype with both theoretical and actual donor-recipient pairs, and the assays were applied to a clinical population of 73 trauma patients who received transfusions where MC was previously characterized by HLA-based assays alone. RESULTS: In the stochastic spiking experiments, all assays were sensitive to a single copy of target DNA, and no false-positive amplification occurred among 1128 samples studied. Among 219 theoretical donor-recipient pairs, informative alleles existed for 99.5 percent with both InDel and HLA compared to 91.3 percent with HLA alone. In the clinical population, 33 cases of MC were detected (9 more cases than by HLA-DR alone) in the nonleukoreduced (non-LR) group and 8 cases (1 more case than by HLA-DR) in the LR group for the short-term follow-up. Among 27 long-term follow-up samples, 8 cases were detected overall (3 more cases than by HLA-DR alone). CONCLUSION: It is concluded that an InDel-based assay panel has excellent technical performance characteristics while also allowing for ascertainment of some MC cases not detectable with HLA alone. The tandem use of both the InDel and the HLA provides a powerful tool for the enhanced ascertainment of MC. PMID- 17076841 TI - A KEL gene encoding serine at position 193 of the Kell glycoprotein results in expression of KEL1 antigen. AB - BACKGROUND: The KEL2/KEL1 (k/K) blood group polymorphism represents 578C>T in the KEL gene and Thr193Met in the Kell glycoprotein. Anti-KEL1 can cause severe hemolytic disease of the fetus and newborn. Molecular genotyping for KEL*1 is routinely used for assessing whether a fetus is at risk. Red blood cells (RBCs) from a KEL:1 blood donor (D1) were found to have abnormal KEL1 expression during evaluation of anti-KEL1 reagents. STUDY DESIGN AND METHODS: Kell genotyping methods, including KEL exon 6 direct sequencing, were applied. KEL cDNA from D1 was sequenced. Flow cytometry was used to assess KEL1 and KEL2 RBC expression. RESULTS: RBCs from the donor, her mother, and an unrelated donor gave weak or negative reactions with some anti-KEL1 reagents. Other Kell-system antigens appeared normal. The three individuals were homozygous for KEL C578 (KEL*2) but heterozygous for a 577A>T transversion, encoding Ser193. They appeared to be KEL*2 homozygotes by routine genotyping methods. Flow cytometry revealed weak KEL1 expression and normal KEL2, similar to that of KEL*2 homozygotes. CONCLUSION: Ser193 in the Kell glycoprotein appears to result in expression of abnormal KEL1, in addition to KEL2. The mutation is not detected by routine Kell genotyping methods and, because of unpredicted KEL1 expression, could lead to a misdiagnosis. PMID- 17076842 TI - Neopterin levels during the early phase of human immunodeficiency virus, hepatitis C virus, or hepatitis B virus infection. AB - BACKGROUND: A study was conducted to assess the diagnostic sensitivity of neopterin screening of blood donors with regard to the detection of window-phase specimens of human immunodeficiency virus (HIV), hepatitis C virus (HCV), and hepatitis B virus (HBV) infection. STUDY DESIGN AND METHODS: In total, 1002 diagnostic window-phase specimens from 98 seroconversion panels (29 HIV-1, 52 HCV, and 17 HBV) were analyzed with viral antigen detection, viral nucleic acid amplification testing (NAT), and neopterin quantitation assays. The study was completed by the analysis of 92 anti-hepatitis B core antigen (HBc)-reactive and 103 alanine aminotransferase (ALT)-elevated blood donor specimens. RESULTS: A significant association between elevated neopterin concentrations and the very early phase of HIV-1 infection was found. No significant correlation could be observed between neopterin levels and the early phase of HCV or HBV infection. Neopterin concentration was not increased in specimens from blood donors with anti-HBc reactivity or ALT elevation. CONCLUSIONS: Neopterin screening of blood donors may identify window-phase cases of HIV, but not of HCV or HBV infection. The diagnostic sensitivity of neopterin screening during the HIV window phase is similar to that of the p24 antigen test. With the introduction of viral NATs in blood screening, there is no additional benefit of neopterin screening with regard to the three blood-borne viruses HIV, HCV, and HBV. Acute phases of other infectious agents, however, have been reported to be detected by neopterin enzyme linked immunosorbent assays. PMID- 17076843 TI - Effects of a new perfluorocarbon emulsion on human plasma and whole-blood viscosity in the presence of albumin, hydroxyethyl starch, or modified fluid gelatin: an in vitro rheologic approach. AB - BACKGROUND: Artificial oxygen carriers such as perfluorocarbon (PFC) emulsions have reached Phase III clinical trials as alternatives to homologous blood, but their rheologic effects have not been characterized. In this study, the rheologic effects of PFC emulsion in the presence of clinically used volume expanders were investigated. STUDY DESIGN AND METHODS: The effects of a new PFC emulsion (small droplet size with narrow size distribution) at two PFC concentrations (4 and 8 g/dL) on plasma and whole-blood viscosity in the presence of human albumin solution (HAS), hydroxyethyl starch (HES), or modified fluid gelatin (MFG) were investigated. Three hematocrit (Hct) levels were investigated: 30, 20, and 13 percent. Plasma, PFC emulsions, and whole-blood viscosity, with a Couette viscometer, and RBC elongation, with an ektacytometer, were measured for shear rates of 0.2 to 128 per second. RESULTS: The two PFC concentrations increased plasma and whole-blood viscosities. Viscosity values similar to physiologic ones (Hct level, 40%) were observed at: 1) Hct level of 13 percent, with 4 or 8 g per dL MFG-PFC; 2) Hct level of 20 percent, with 4 g per dL MFG-PFC; and 3) Hct level of 30 percent, with 4 g per dL HES-PFC and 4 and 8 g per dL HAS-PFC. RBC deformability was unchanged. CONCLUSION: It is concluded that this new PFC emulsion increases plasma and blood viscosity and that among the three studied volume expanders, the interaction with MFG can result in viscosity values above the physiologic one even at low Hct values. The possible consequences of the increased viscosity at low Hct values are discussed. PMID- 17076844 TI - Acute transfusion reactions in the pediatric intensive care unit. AB - BACKGROUND: Acute transfusion reactions (ATRs) are probably underdiagnosed in critically ill children because associated symptoms can frequently be attributed to the patient's underlying disease. This study was undertaken to determine the incidence, type, imputability, and severity of ATRs observed in a tertiary care pediatric intensive care unit (PICU). STUDY DESIGN AND METHODS: All transfusions of labile blood product administered to consecutive patients admitted to our PICU, between February 2002 and February 2004, were prospectively recorded. For each transfusion, the bedside nurse recorded the patient's status before, during, and up to 4 hours after the transfusion, as well as the presence of any new sign or symptom suggesting an ATR. Three independent experts retrospectively reviewed all transfusion event reports and hospital charts. The presence, type, imputability, and severity of ATRs were adjudicated by consensus of two of three experts (Delphi method), with predefined criteria. RESULTS: A total of 2509 transfusions were administered to 305 patients during the study. Forty transfusion events (1.6%) were confirmed to be ATRs by expert consensus: 24 febrile nonhemolytic, 6 minor allergic, 4 isolated hypotension, 3 bacterial contamination, 1 major allergic (anaphylactic shock), 1 TRALI, and 1 hemolytic reaction. Imputability of ATRs was probable or possible in 35 cases (88%). ATRs led to an immediate vital threat in 15 percent of cases. CONCLUSION: Improved surveillance of transfusions given to PICU patients and better knowledge of these reactions by health care professionals should improve the safety of transfusions in the PICU. PMID- 17076845 TI - Safety and efficacy of therapeutic early onset granulocyte transfusions in pediatric patients with neutropenia and severe infections. AB - BACKGROUND: Bacterial and fungal infections in profound neutropenia after chemotherapy are associated with high mortality despite appropriate antibacterial and antifungal treatment. Granulocyte transfusions are used as a therapeutic addendum, but concern regarding pulmonary reactions often results in delayed use in clinical practice. Accordingly, many patients are already at advanced stages of their infectious disease once granulocytes are transfused. Thus, a prospective Phase II trial was conducted to test the safety and efficacy of therapeutic early onset granulocyte transfusions in immunocompromised children with neutropenia and severe infections. STUDY DESIGN AND METHODS: Twenty-seven children with hematologic disorder or malignancy and severe neutropenia with clinically and/or microbiologically documented severe infection unresponsive to standard treatment were included. They received granulocyte colony-stimulating factor (G-CSF) elicited, crossmatched granulocyte concentrates every other day until complete recovery from infection was documented. RESULTS: A median of two granulocyte transfusions with a median of 8 x 10(8) granulocytes per kilogram of body weight were administered. All transfusions were well tolerated, and no pulmonary symptoms were observed. A total of 92.6 percent of our patients were able to clear their initial infection, and 81.5 percent were alive and without signs or symptoms of their infection 1 month later. All six children with aspergillosis cleared their infection. CONCLUSIONS: G-CSF-elicited, crossmatched granulocyte concentrates are a safe and efficient therapeutic addendum in immunocompromised children with prolonged neutropenia and severe infections. Early transfusion of granulocyte concentrates can lead to an overall response rate of 92.6 percent without adverse events. Randomized clinical trials with an early-onset design are required to determine appropriate clinical applications. PMID- 17076846 TI - Estimating blood needs for very-low-birth-weight infants. AB - BACKGROUND: Red blood cell (RBC) transfusions are crucial for the care of very low-birth-weight (VLBW) infants. These infants frequently require multiple, small volume RBC transfusions, with potential exposure to multiple donors. Optimal protocols provide dedicated RBC units to reduce exposures and avoid RBC wastage. STUDY DESIGN AND METHODS: This study was a retrospective, single-institution review of RBC transfusions in VLBW infants. The RBC volume transfused during the entire hospitalization (VTH) and that transfused at 35 days were determined for all infants, 401 to 1250 g at birth, admitted to a Level III neonatal intensive care unit from January 1, 2000, through December 31, 2002. Multivariable models identified perinatal factors associated with volume transfused. RESULTS: The 640 infant cohort had a median birth weight (BW) of 818 g and gestational age (GA) of 26 weeks. Most infants (546 or 85%) required at least one RBC transfusion. Median number of RBC transfusions was 3 (range, 0-30) and median volume transfused was 82 mL (range, 9-737 mL). Of 328 infants who received all transfusions within a 35 day period, only 5 (1.5%) required at least 200 mL. VTH was inversely related to BW and GA. Multivariable models identified BW, GA, age at first transfusion, and use of inotropes as variables associated with higher volume transfused. CONCLUSION: Few VLBW infants use an entire RBC unit. One dedicated unit shared by two or more infants should meet their transfusion needs. GA, BW, and markers of illness severity predict increased RBC volume requirements. PMID- 17076847 TI - Population-based audit of fresh-frozen plasma transfusion practices. AB - BACKGROUND: In contrast to decreasing red blood cell (RBC) consumption in Finland, the use of fresh-frozen plasma (FFP) has been increasing since the 1990s, suggesting that FFP use may not always be optimal. To improve transfusion practices, knowledge of current FFP use and regional, national, and international comparison is necessary. STUDY DESIGN AND METHODS: Nine (of 21) Finnish hospital districts participated. Data concerning FFP-transfused patients in the years 2002 and 2003 were collected from existing computerized medical records into a yearly updated database as part of a Finnish benchmarking project on blood component use. RESULTS: Data included 11,590 FFP-transfused patients and 60,240 FFP units (71.2% of Finnish FFP use) delivered to Finnish hospitals during the study period. FFP was transfused most often to surgery patients (62.8% of FFP transfusion hospital visits) with blood circulatory system problems (32.3% of surgically treated and FFP-transfused patients). In only 65.9 percent of FFP transfused patients were coagulation variables measured at any point in the hospital episode, and FFP was usually transfused in paired doses. Mean FFP use in Finland is comparable to other countries. CONCLUSION: Although overall FFP use in Finland is similar to that of international figures, it does not ensure best practice. Perioperative staff, being the largest FFP user, should be encouraged to dose FFP based on coagulation variables and body weight. Improvement efforts should be directed to patient groups transfused with large amounts of FFP. PMID- 17076848 TI - A study of the agreement between patient self-assessment and study personnel assessment of bleeding symptoms. AB - BACKGROUND: Clinical trials investigating new platelet transfusion therapies frequently require the assessment of bleeding for the study outcome. These assessments are commonly performed by study personnel and can be time-consuming. The purpose of this study was to assess whether patients were able to reliably assess their bleeding status on a daily basis. STUDY DESIGN AND METHODS: Patients admitted to hospital to receive chemotherapy for acute leukemia or to undergo allogeneic peripheral blood progenitor cell transplant were included. Patients were given an introduction to a form for documenting the occurrence of 16 bleeding symptoms. Patients completed this form and were examined daily by a study assessor. A weekly health record review was also performed by a study assessor. The agreement between raters was determined by calculating the raw agreement, chance-corrected agreement, and chance-independent agreement. RESULTS: Thirty-five patients completed 458 assessment forms that were paired with 559 forms completed by a study assessor with 450 matched forms available for analysis (mean, 12.86 per patient). Agreement for most individual bleeding symptoms was high. Thirteen items had agreement greater than 90 percent and all items had agreement greater than 77 percent. The lowest agreement was seen for skin symptoms: petechiae (89.2%), purpura (80.9%), and ecchymosis (77.6%). The negative predictive value of patient self-assessment was high (range, 71.1%-100%) whereas the positive predictive value was lower (range, 0%-86.5%). CONCLUSION: The reliability was very good between patients and study assessors with the patients reporting excellent negative predictive value and variable positive predictive value. PMID- 17076849 TI - Large-scale expansion and transplantation of CD34(+) hematopoietic cells: in vitro and in vivo confirmation of neutropenia abrogation related to the expansion process without impairment of the long-term engraftment capacity. AB - BACKGROUND: Herein are reported the results obtained in all multiple myeloma patients transplanted with peripheral blood hematopoietic progenitor cells submitted to ex vivo expansion. STUDY DESIGN AND METHODS: Patients had blood progenitor cell mobilization with cyclophosphamide and filgrastim. CD34+ cells were expanded for 10 days in a medium containing granulocyte-colony-stimulating factor (G-CSF), stem cell factor, and megakaryocyte growth and development factor (MGDF). Twenty-seven patients underwent transplantation with expanded and nonexpanded cells and 7 patients underwent transplantation with expanded cells only. RESULTS: The median fold cell expansion was 29.1. The number of colony forming unit-granulocyte-macrophage (CFU-GM) and CD34+ cells, and the long-term culture-initiating cell (LTC-IC) activity increased with median fold values of 14.7, 2.75, and 2.25, respectively. Postmyeloablative neutropenia was abrogated in 24 of 27 patients transplanted with expanded cells plus nonexpanded cells. The median duration of severe neutropenia was 0 days and correlated with the number of cells and CFU-GM infused. Survival was similar to that of a historical control group. Our LTC-IC and NOD-SCID mice studies showed that the expanded cells are able of sustaining long-term hematopoiesis. Seven other patients received transplantation with expanded cells alone. Absolute neutropenia was abrogated in 6 patients. The median duration of neutropenia was 0 days. Two patients who received the lower number of total cells or CFU-GM had brief secondary neutropenia, which resolved after G-CSF injections. CONCLUSION: CD34+ cells expanded ex vivo can abrogate absolute and severe neutropenia after high-dose therapy. The results of the amplification process are strongly related to the delay of hematopoietic recovery. PMID- 17076850 TI - Primary and secondary hemostatic functionalities of rehydrated, lyophilized platelets. AB - BACKGROUND: The rehydrated, lyophilized (RL) platelet (PLT) is being developed as a hemostatic infusion agent for the control of active bleeding. The key to the method for preparing RL PLTs is a mild aldehyde stabilization that allows for freezing and lyophilizing without cellular rupture. RL PLTs have been shown to be effective at rapidly controlling bleeding in animal models of cardiopulmonary bypass induced PLT dysfunction and washout thrombocytopenia, yet the rehydrated cells have proved to be safe with respect to induction of pathologic intravascular coagulation. STUDY DESIGN AND METHODS: In vitro and in vivo studies were performed to better understand the differential effect of the RL PLT manufacturing method on primary and secondary hemostatic processes. The functionality of the von Willebrand factor (VWF) receptor (glycoprotein Ib) complex, the PAR receptors, integrin-mediated aggregation (inside-out signaling), and surface membrane prothrombin to thrombin conversion systems were investigated. RESULTS: RL PLTs were found to retain native VWF-mediated adhesion and surface thrombin generation functions. In contrast, the coupling of thrombin receptors to integrin inside-out signaling was largely inhibited. CONCLUSION: These results suggest that RL PLTs may stop bleeding by forming primary hemostatic plugs and providing a localized source of thrombin for secondary hemostatic processes, yet do not build up occlusive pathologic clots possibly because integrin functions for forming PLT-PLT aggregates are partially inhibited. PMID- 17076851 TI - Resistance of porcine circovirus and chicken anemia virus to virus inactivation procedures used for blood products. AB - BACKGROUND: Virus inactivation procedures are used to prevent contamination of plasma-derived blood products with viruses. Pasteurization or prolonged dry heat has proven effective against several enveloped and nonenveloped viruses and provides an additional layer of safety for plasma products. STUDY DESIGN AND METHODS: The resistance of porcine circovirus 2 (PCV2) and chicken anemia virus (CAV), two small, nonenveloped viruses, to standard (pasteurization, 10 hr at 60 degrees C; dry heating, 80 degrees C for 72 hr) and more extreme heat inactivation procedures (temperatures up to 120 degrees C) was determined. The ability of these procedures to inactivate PCV2 and CAV was measured by comparison of in vitro infectivity before and after treatment. RESULTS: Infectivity of PCV2 and CAV was reduced by approximately 1.6 and 1.4 log by pasteurization and by 0.75 and 1.25 log by dry-heat treatment, both substantially more resistant than other viruses previously investigated. PCV2 and CAV were additionally almost completely resistant to dry-heat treatment up to 120 degrees C for 30 minutes (mean log infectivity reductions, 1.25 and 0.6), although both were more effectively inactivated when the temperature of wet-heat treatment was increased to 80 degrees C (>3.2 and >3.6 log infectivity reduction). CONCLUSION: Although neither PCV2 nor CAV are known to infect humans, their inactivation properties may represent those of other small DNA viruses known to be present (e.g., TT virus, small anellovirus) or potentially present in human plasma. Findings of extreme thermal resistance demonstrate that recipients of plasma-derived therapeutics may potentially still be exposed to small DNA viruses, despite the implementation of viral inactivation steps. PMID- 17076852 TI - Biochemical, molecular characterization, and glycoproteomic analyses of alpha(1) proteinase inhibitor products used for replacement therapy. AB - BACKGROUND: Isoelectric focusing (IEF) of alpha(1)-proteinase inhibitor (A1PI) shows that commercial products and plasma have different glycoisoform band patterns. Those in Aralast (Grifols Biologicals) reflect an anodal shift of glycoisoforms, which has caused concern. The protein, including glycoproteomic analyses, and structural features of A1PI products were investigated by state-of the-art techniques. STUDY DESIGN AND METHODS: Batches from Aralast, Prolastin (Bayer), and Zemaira (Aventis Behring LLC) were analyzed by high-resolution IEF and high-performance size-exclusion chromatography (HP-SEC). Preparative separated isoforms from IEF were further purified by chromatography and subjected to mass spectrometry for sequence analyses, peptide mapping, and glycosylation analysis. Deamidation was quantified by enzymatic isoaspartate detection. Multiple sequence alignments and structural bioinformatics analyses were performed. RESULTS: In HP-SEC, Prolastin had the highest aggregate content at approximately 30 percent. Isoforms from all products purified by high-resolution IEF were sequenced with an amino acid coverage of more than 98 percent. Deamidation of Asn116 and Asn314 in A1PI was to found to some extent in all products and confirmed quantitatively by enzymatic analysis. There were no signs of methionine oxidation. Cys232 was found to be cysteinylated in A1PI in Prolastin and Aralast as in plasma, but not in Zemaira. All products showed truncation of the C-terminal lysine. Intact A1PI concentrates contained mainly diantennary, disialylated and smaller amounts of triantennary, trisialylated N glycans. The percentage of fucosylation was similar in all products. Site specific glycan analysis revealed bands M6 contained only diantennary glycans, whereas the more acidic bands M4 and M2 also carried triantennary structures. The most acidic isoforms, M2 in Prolastin and Zemaira and M0 in Aralast, additionally exhibited tetraantennary N-glycans. CONCLUSION: Protein chemical characterization of A1PI showed that all A1PI products to some extent differ from A1PI circulating in human plasma. Bioinformatic analysis indicated that removal of C-terminal Lys394 and cysteinylation of Cys232 are unlikely to affect structure and/or function of A1PI but cysteinylation may influence interaction between A1PI and its physiologic ligands. Aralast, Prolastin, and Zemaira contain the same set of N-glycans in the same ratios as those in normal human plasma A1PI. Tri- and tetraantennary structures are responsible for the partitioning into IEF isoforms, with the migration shift of Aralast not being due to any difference in the N glycosylation, but to the partial loss of the C-terminal lysine. PMID- 17076853 TI - Dispermic chimerism identified during blood group determination and HLA typing. AB - BACKGROUND: Chimerism is the presence of two or more genetically distinct cell populations in one organism. STUDY DESIGN AND METHODS: We report the identification of dispermic chimerism in a 19-year-old female volunteer blood donor. During routine ABO blood grouping strong reactions of the blood donors red blood cells (RBCs) with anti-A reagents and mixed-field reactions with anti-B reagents were observed, while serum-testing showed the absence of anti-A and anti B antibodies. AB0 blood group genotyping, HLA-typing and microsatellite analysis were performed using blood-samples, buccal mucosa and fibroblasts of the blood donor and blood-samples of her parents. RESULTS: AB0 blood group genotyping showed three ABO blood group alleles (0(1), A(2) and B) in the DNA-samples of the blood-donor. The evidence of chimerism was supported by the detection of three alleles for the HLA-A and HLA-DRB1 loci. Microsatellite analysis with ten markers revealed three alleles for loci D7S821 and D19S412. All studies carried out, the third allele was always of paternal origin. CONCLISION: The results suggested a case of a human dispermic chimerism. Our proposed explanation for the development of chimerism in the reported case is the fertilization of an oocyte and the corresponding second polar body by two different sperms. PMID- 17076854 TI - A novel IGnT allele responsible for the adult i phenotype. AB - BACKGROUND: The adult i phenotype has been characterized as the presence of a very low level of I antigen but a high quantity of I antigen on red blood cells (RBCs). It has been noted that this rare phenotype is partially associated with congenital cataracts. It has been demonstrated that the human I locus expresses three IGnT forms, IGnTA, IGnTB, and IGnTC, and that the IGnTC gene is responsible for the I antigen expression on RBCs. This report describes molecular genetic analysis of a Taiwanese person with the adult i phenotype but without congenital cataracts. STUDY DESIGN AND METHODS: The five exon regions of the IGnT gene of the adult i individual were amplified by polymerase chain reaction (PCR) and cloned, and the sequences were determined. The activity of the IGnT enzyme expressed from the mutant IGnTC gene identified in this i adult was analyzed. RESULTS: The presented adult i individual possesses wild-type IGnTA and IGnTB genes but a mutant IGnTC gene with a 243T>A nucleotide substitution, which predicts an amino acid alteration of Asn81Lys. PCR-restriction fragment length polymorphism analysis has been used to show that this IGnTC*243A allele is uncommon in the general Taiwanese population. The activity of the IGnT enzyme expressed from the mutant IGnTC*243A gene was significantly reduced when compared with that expressed from the wild-type IGnTC gene. CONCLUSION: A novel IGnTC allele with a 243T>A missense mutation was demonstrated in our adult i Taiwanese without congenital cataracts. The molecular basis revealed for this adult i case agrees with the proposed molecular genetic mechanism, accounting for the partial association of the adult i phenotype with congenital cataracts. PMID- 17076855 TI - Expression of the histo-blood group B gene predominates in AB-genotype cells. AB - BACKGROUND: It has been demonstrated that the 43-bp minisatellite sequence in the 5' region of the ABO gene plays an important role in its transcriptional regulation. It was determined in previous investigations that the structure of the minisatellite enhancer was specific to A, B, and O alleles. STUDY DESIGN AND METHODS: Real-time polymerase chain reaction (PCR) detection and a PCR restriction fragment length polymorphism (RFLP) strategy were used to compare the quantities of the A and B transcripts in AB-genotype cells, including peripheral blood cells and cancer cell line with the group AB phenotype. The 5' 3.7-kb regions of the A and B genes were cloned and the sequences compared. The transcriptional activities of the 5' segments of the A and B genes were compared with luciferase reporter assay. RESULTS: Both real-time PCR and PCR-RFLP analyses show that there is evidently more of the B transcript in the AB-genotype cells. It was demonstrated that the 5' segment of the B gene had a markedly higher transcription-activation activity relative to the A gene. This difference in transcription capability appears to result from the variation in minisatellite enhancer structures in the A and B genes, which contain one and four repeats of the 43-bp enhancer unit, respectively. CONCLUSION: Our study indicates that the majority of steady-state mRNA within AB-genotype cells is composed of the B transcript and that this phenomenon is due to the predominant expression of the B gene relative to the A gene. PMID- 17076856 TI - Prevalence of selected viral infections among blood donors deferred for potential risk to blood safety. AB - BACKGROUND: Health history questions identify blood donors believed to pose a higher risk of transmission of infectious diseases. This study assesses the current impact of some of these questions on blood safety as reflected by infectious disease markers. STUDY DESIGN AND METHODS: Donors who were deferred from donating blood due to health history question(s) were recruited at four different regions of the American Red Cross Blood Services. A blood sample was tested for serologic markers of blood-borne infections as performed for accepted blood donors. RESULTS: Of 497 deferred donors enrolled, 29 donors were deferred for having had "yellow jaundice, liver disease, or hepatitis since the age of 11" (Question 3), 1 of whom had hepatitis C virus antibodies (anti-HCV) and hepatitis B core antigen antibodies (anti-HBc), 2 had anti-HBc, and 1 had anti-HCV (p < 0.05 for both markers). Among 37 donors deferred for having "ever tested positive for hepatitis" (Question 4), 1 had hepatitis B surface antigen and anti-HBc and 3 had anti-HBc (p < 0.05 for both markers). Of 14 donors deferred for "having ever used a needle, even once, to take any illegal or nonprescription drug" (Question 12), 1 had anti-HCV, human T-lymphotropic virus-I antibodies and anti-HBc, 1 had anti-HCV and anti-HBc, and 2 had anti-HCV (p < 0.05 for all three markers). CONCLUSIONS: Blood donors deferred for standard blood donor questions regarding risk of viral hepatitis as well as those with a history of intravenous drug use were more likely to have higher hepatitis marker rates than those who were not deferred. No significant findings were identified for other markers or questions. PMID- 17076857 TI - Prospective, paired crossover comparison of multiple, single-needle plateletpheresis procedures with the Amicus and Trima Accel cell separators. AB - BACKGROUND: The Baxter Amicus Version 2.51 (A) and the Gambro BCT Trima Accel Version 5.0 (T) cell separators may produce multiple platelet (PLT) concentrates within a single donation. STUDY DESIGN AND METHODS: The single-needle multiple plateletpheresis procedures of the two devices were compared in a prospective, randomized, paired crossover study in 60 donors. The 120 donations were compared for donor comfort, collection efficiency, residual white blood cell (WBC) count, and (in selected patients) corrected count increment (CCI). RESULTS: The mean PLT yield and the resultant mean number of units per donation were significantly lower for A (6.06 x 10(11) vs. 7.48 x 10(11) and 2.57 vs. 3.19, respectively, both p < 0.001), in spite of a longer apheresis duration (89 min vs. 79 min; p < 0.001). This resulted in a higher collection rate of T (5.68 x 10(11) PLTs/hr vs. 4.10 x 10(11) PLTs/hr, p < 0.001). Residual WBC count of every unit was fewer than 5 x 10(6), but significantly fewer A-PLT donations contained more than 10(5) WBCs per unit (1 vs. 9, p = 0.008). Although the ACD-A consumption was slightly higher for A (489 mL vs. 469 mL, p = 0.04), a trend to a higher frequency of side effects was found for T (42.4% vs. 23.7%, p = 0.06). The 1-hour CCIs of 33 transfused A-PLT units were comparable with those of 43 T-PLT units (11.8 vs. 13.9, p = 0.480). CONCLUSIONS: Both cell separators showed safe collections of up to 4 PLT units per donation with adequate CCI. T produced a higher PLT yield despite shorter apheresis duration, but with slightly higher residual WBC counts and a trend to a higher side-effect frequency. PMID- 17076858 TI - Managing threats rather than risks in blood transfusion: robust design for a complex system. PMID- 17076859 TI - Clinical consequences of red cell storage in the critically ill. AB - Red cell (RBC) transfusions are a potentially life-saving therapy employed during the care of many critically ill patients to replace losses in hemoglobin to maintain oxygen delivery to vital organs. During storage, RBCs undergo a series of biochemical and biomechanical changes that reduce their survival and function. Additionally, accumulation of other biologic by-products of RBC preservation may be detrimental to recipients of blood transfusions. Laboratory studies and an increasing number of observational studies have raised the possibility that prolonged RBC storage adversely affects clinical outcomes. In this article, the laboratory and animal experiments evaluating changes to RBCs during prolonged storage are reviewed. Subsequently, the clinical studies that have evaluated the clinical consequences of prolonged RBC storage are reviewed. These data suggest a possible detrimental clinical effect associated with the transfusion of stored RBCs; randomized clinical trials further evaluating the clinical consequences of transfusing older stored RBCs are required. PMID- 17076860 TI - Individual nucleic amplification technology does not prevent all hepatitis B virus transmission by blood transfusion. PMID- 17076861 TI - Are weak D red blood cells really immunogenic? PMID- 17076863 TI - Reactions to blood donation in Caucasian teenagers. PMID- 17076866 TI - Impact of rofecoxib withdrawal on cyclooxygenase-2 utilization among patients with and without cardiovascular risk. AB - OBJECTIVE: To compare how cyclooxygenase-2 (COX-2) users with and without cardiovascular (CV) risks responded to the withdrawal of rofecoxib on September 30, 2004. METHODS: The data, from a pharmacy claims database, consisted of patients who filled at least one prescription for a COX-2 agent in the 6 months before September 30, 2004 (n = 32,898) or September 30, 2003 for the control cohort (n = 37,930). Baseline drug utilization was used to determine comorbidities, and CV risk status was assessed with surrogate pharmacy markers. The first difference estimator was used to compare changes in utilization after September 30, 2004 with changes in utilization in a control cohort of patients who were treated with COX-2 over a similar time frame in 2003/2004. RESULTS: The reduction in COX-2 utilization depended on baseline CV risk status. Among celecoxib and valdecoxib users, patients without CV risks reduced their utilization of COX-2, as measured by days of supply, by between 16.2% and 22.7%. The reduction was 32% for patients with one CV risk marker and 55.8% for patients with three or more markers, a proxy for the severity of CV risk. The corresponding figures for rofecoxib users were 47.5% (no CV risk), 55.4% (one marker) and 64.8% (three or more markers). CONCLUSIONS: Our results suggest that patients and physicians used newly available information about COX-2 inhibitor agents and their side effects to reduce treatment. They also provide support for the notion that patients and providers were discriminating in their response to new information as a significant proportion of patients remained on treatment after extensive publicity concerning potential risks. PMID- 17076867 TI - Estimating the cost-effectiveness of quality-improving interventions in oral anticoagulation management within general practice. AB - OBJECTIVES: A clinical trial, "Belgian Improvement Study on Oral Anticoagulation Therapy (BISOAT)," significantly improved the quality after implementing four different quality-improving interventions in four randomly divided groups of general practitioners (GPs). The quality-improving interventions consisted of multifaceted education with or without feedback reports on their performance, international normalized ratio (INR) testing by the GP with a CoaguChek device or computer-assisted advice for adapting oral anticoagulation therapy. The quality improvement in INR control versus baseline was similar in the four groups. The aim of the current study was to calculate the cost-effectiveness and influencing factors of the four quality-improving interventions compared with usual care. METHODS: Activity-based costing techniques with questionnaires were used to determine the global costs per patient per month in the different intervention groups. Effectiveness data were obtained from the BISOAT study. Cost effectiveness was expressed as cost per additional day within a 0.5 range from INR target. RESULTS: The one-time cost of multifaceted education was 49,997 euro for the whole study. Monthly continuous costs per intervention ranged between 37 euro and 54 euro per patient. Using the CoaguChek in combination with the multifaceted education was associated with net savings and quality improvement, hence dominated usual care. Sensitivity analyses showed improved cost effectiveness with extended duration and with increased program size. CONCLUSION: Implementation of the combination multifaceted education with the use of the CoaguChek is a cost-effective new organizational model of oral anticoagulation management in general practice. PMID- 17076868 TI - Too much ado about propensity score models? Comparing methods of propensity score matching. AB - OBJECTIVE: A large number of possible techniques are available when conducting matching procedures, yet coherent guidelines for selecting the most appropriate application do not yet exist. In this article we evaluate several matching techniques and provide a suggested guideline for selecting the best technique. METHODS: The main purpose of a matching procedure is to reduce selection bias by increasing the balance between the treatment and control groups. The following approach, consisting of five quantifiable steps, is proposed to check for balance: 1) Using two sample t-statistics to compare the means of the treatment and control groups for each explanatory variable; 2) Comparing the mean difference as a percentage of the average standard deviations; 3) Comparing percent reduction of bias in the means of the explanatory variables before and after matching; 4) Comparing treatment and control density estimates for the explanatory variables; and 5) Comparing the density estimates of the propensity scores of the control units with those of the treated units. We investigated seven different matching techniques and how they performed with regard to proposed five steps. Moreover, we estimate the average treatment effect with multivariate analysis and compared the results with the estimates of propensity score matching techniques. The Medstat MarketScan Data Base provided data for use in empirical examples of the utility of several matching methods. We conducted nearest neighborhood matching (NNM) analyses in seven ways: replacement, 2 to 1 matching, Mahalanobis matching (MM), MM with caliper, kernel matching, radius matching, and the stratification method. RESULTS: Comparing techniques according to the above criteria revealed that the choice of matching has significant effects on outcomes. Patients with asthma are compared with patients without asthma and cost of illness ranged from 2040 dollars to 4463 dollars depending on the type of matching. After matching, we looked at the insignificant differences or larger P-values in the mean values (criterion 1); low mean differences as a percentage of the average standard deviation (criterion 2); 100% reduction bias in the means of explanatory variables (criterion 3); and insignificant differences when comparing the density estimates of the treatment and control groups (criterion 4 and criterion 5). Mahalanobis matching with caliber yielded the better results according all five criteria (Mean = 4463 dollars, SD = 3252 dollars). We also applied multivariate analysis over the matched sample. This decreased the deviation in cost of illness estimates more than threefold (Mean = 4456 dollars, SD = 996 dollars). CONCLUSION: Sensitivity analysis of the matching techniques is especially important because none of the proposed methods in the literature is a priori superior to the others. The suggested joint consideration of propensity score matching and multivariate analysis offers an approach to assessing the robustness of the estimates. PMID- 17076869 TI - Longitudinal metric properties of disability rating scales for Parkinson's disease. AB - OBJECTIVES: This study analyzes the longitudinal metric attributes of three Parkinson's disease (PD) disability scales, taking Hoehn and Yahr (HY) staging as the reference measure of PD progression. METHODS: A sample of 87 PD patients was assessed during regular medical visits, using the HY, the Unified Parkinson's Disease Rating Scale--Activities of Daily Living Section (UPDRS-ADL), the Schwab and England Scale (SES), and the Intermediate Scale for Assessment of PD (ISAPD), across a follow-up period of 2.6 +/- 1.0 years. RESULTS: The following cross sectional attributes were analyzed, at baseline and again on conclusion of the study: floor and ceiling effects, convergent validity, reliability, and standard error of measurement, all of which were found to be adequate. Longitudinal reproducibility values (intraclass correlation coefficient) were 0.81 (ISAPD) to 0.84 (UPDRS-ADL). Insofar as longitudinal validity was concerned, the change scores of the three disability scales correlated significantly with the HY change score, absolute value r = 0.33 to 0.45, P < 0.003. Slightly lower values were found when taking the annual rate of change, absolute value r = 0.20 to 0.36. The three scales were acceptable, even though there were small differences among them. The "minimal clinically important difference" proposed for these scales is: SES, -6; UPDRS-ADL, +2; ISAPD, +1.5 points. CONCLUSIONS: The three scales proved adequate for longitudinal assessment of PD disability. UPDRS-ADL was more precise and ISAPD more consistent. Magnitude of change and correlation with change in HY were slightly higher with the ISAPD. Effect size and standardized response mean for the minimal change in HY were higher for the UPDRS-ADL. PMID- 17076870 TI - Integrating the patient's perspective into device evaluation trials. AB - Innovations in medical device technology have greatly expanded the range of therapeutic options available to physicians and their patients. The understanding of treatment effects from the patient's perspective is an essential component of a comprehensive assessment of any new therapy, including medical devices. The term "patient-reported outcomes" (PROs) has been growing in use to refer to a cluster of variables such as health-related quality of life, symptoms, physical functioning, psychological well-being, treatment satisfaction, and treatment preferences. As in drug trials, the use of PROs in device evaluation has several methodological challenges, ranging from general concerns about interpretation, to more specific issues related to study design and regulatory approval (use of PROs as primary end points, incorporation in labeling, and product promotion). Successful approaches for integrating PROs into device evaluation trials include the careful selection of appropriate, interpretable PRO end points, accounting for possible confounding factors, and the use of alternatives to placebo controlled trial designs, such as single-arm pre-post, observational, and registry studies, when the use of placebo control groups is not feasible. This article discusses the potential value and difficulties in measuring PROs in device studies. PMID- 17076871 TI - Immunohistochemical properties of ocular adenomas, adenocarcinomas and medulloepitheliomas. AB - Ocular medulloepitheliomas, adenomas and adenocarcinomas share a common phenotype and originate from the optic cup neuroectoderm. This can make it very difficult to differentiate between these tumors histopathologically. Therefore, this study focused on identifying a combination of immunologic markers that might be used in the diagnosis of these tumors. These markers included AE1/AE3, CK7, CK20, and telomerase reverse transcriptase (TERT). Routine immunohistochemical staining was performed on 27 whole globes diagnosed with one of these tumors. The tumors that immunostained for TERT showed increasing immunoreactivity as the tumor types increased in aggressiveness. None of the tumor types were immunopositive for CK7. CK20 immunostaining was found in the adenomas but not in the adenocarcinomas or medulloepitheliomas. AE1/AE3 expression was present more consistently in the adenocarcinomas and less frequently in the adenomas. AE1/AE3 expression was present in only one of six medulloepitheliomas. Furthermore, CK20 and TERT showed inverse expression patterns, i.e. TERT increased in expression and CK20 decreased in expression with increasing aggressiveness. These results may be important diagnostic and prognostic indicators for these tumors. PMID- 17076872 TI - Classification of feline intraocular neoplasms based on morphology, histochemical staining, and immunohistochemical labeling. AB - The objectives of this study were to evaluate morphologic, histochemical, and immunohistochemical characteristics of well-differentiated and anaplastic intraocular neoplasms of cats, and to develop a diagnostic algorithm for, and investigate the association of ruptured lenses with these neoplasms. Seventy-five feline globes with intraocular neoplasms were stained with hematoxylin and eosin and examined by light microscopy. Morphologic diagnoses included 33 intraocular sarcomas, 17 diffuse iris melanomas, 15 lymphosarcomas, three ciliary adenomas, one metastatic carcinoma, and six undifferentiated intraocular neoplasms. Sections of these globes were then stained with periodic acid Schiff (PAS), and immunohistochemical (IHC) labels for various cellular markers. Histochemical staining and IHC labeling confirmed cellular differentiation in 73/75 neoplasms but was discordant with morphologic diagnoses in 8/75. These included four neoplasms morphologically diagnosed as lymphosarcomas but which expressed differentiation antigens consistent with melanoma (n = 3) or ciliary adenocarcinoma (n = 1), and four tumors morphologically diagnosed as intraocular sarcomas that expressed differentiation antigens for melanoma (n = 2), metastatic carcinoma (n = 1), or remained undifferentiated (n = 1). Immunohistochemical labeling suggested a diagnosis in 5/6 morphologically undifferentiated neoplasms including one intraocular sarcoma, two diffuse iridal melanomas, and two ciliary adenocarcinomas. Based upon morphologic, histochemical, and IHC characterization, ruptured lens capsules were detected in 28/30 intraocular sarcomas, 3/24 diffuse iris melanomas and 1/11 lymphosarcomas, but not in ciliary epithelial neoplasms, metastatic carcinomas, or undifferentiated intraocular neoplasms. An algorithm is provided that facilitates stain and IHC label selection for differentiating anaplastic intraocular feline neoplasms. PMID- 17076873 TI - Amniotic membrane transplantation for corneal surface reconstruction after excision of corneolimbal squamous cell carcinomas in nine horses. AB - OBJECTIVE: The purpose of this study was to evaluate the usefulness and effectiveness of permanent amniotic membrane transplantation as an adjunctive treatment to superficial keratectomy alone or combined with strontium-90 irradiation for treatment of equine corneolimbal squamous cell carcinoma (SCC) to decrease corneal scarring and recurrence rate. STUDY: The retrospective case study included 11 horses (n = 12 eyes) diagnosed and treated for ocular SCC that involved the limbus and cornea. Nine of those horses (n = 9 eyes) were treated between 2002 and 2006, with superficial lamellar keratectomy alone or combined with strontium-90 irradiation and followed by placement of a permanent amniotic membrane graft in the surgical defect. The level of scarring (i.e. the clarity of the cornea) resulting with the use of amniotic membrane was subjectively compared to cases where a permanent bulbar conjunctival graft was performed following keratectomy combined with strontium-90 irradiation or cryotherapy (n = 3 eyes). Recurrence was defined as the postoperative and postirradiation regrowth of SCC in the same site and globe. RESULTS: The nine horses that received an amniotic membrane graft after keratectomy alone or combined with irradiation showed a minimal level of scarring in a cornea that regained a greater transparency in comparison to the horses that were treated with a bulbar conjunctival graft. All of the horses that received an amniotic membrane graft had 226 +/- 218 days of follow-up without tumor recurrence (mean +/- SD), ranging from 21 days to 778 days. CONCLUSIONS: The combination of superficial keratectomy alone or associated with beta-irradiation and permanent amniotic membrane transplantation is an effective treatment of corneal or corneolimbal SCC in horses. The placement of an amniotic membrane material represents an alternative surgical procedure to bulbar conjunctival grafts, especially if there is a lack of bulbar conjunctiva tissue available after tumor resection or if a particularly large corneal resection is necessary. The amniotic membrane is incorporated into the corneal defect and seems to create noticeably much less scarring than a corneal defect covered by bulbar conjunctiva. PMID- 17076877 TI - High-throughput genomic technology in research and clinical management of breast cancer. Exploiting the potential of gene expression profiling: is it ready for the clinic? AB - Gene expression profiling is a relatively new technology for the study of breast cancers, but within the past few years there has been a rapid rise in interest in its potential to improve the clinical management of breast cancer. This technology has contributed to our knowledge of the molecular pathology of breast tumours and shows promise as a tool to predict response to therapy and outcome, such as risk of metastasis. Microarray technology is continually developing and it is becoming apparent that, despite the various platforms available, robust conclusions can still be drawn that apply across the different array types. Gene expression profiling is beginning to appear in the breast cancer clinic but it is not yet fully evaluated. This review explores the questions that must be addressed before this technology can become an everyday clinical tool. PMID- 17076878 TI - Leaner and meaner genomes in Escherichia coli. AB - A 'better' Escherichia coli K-12 genome has recently been engineered in which about 15% of the genome has been removed by planned deletions. Comparison with related bacterial genomes that have undergone a natural reduction in size suggests that there is plenty of scope for yet more deletions. PMID- 17076879 TI - A Melanesian alpha-thalassemia mutation suggests a novel mechanism for regulating gene expression. AB - A Melanesian variant of the genetic disease alpha-thalassemia has recently been shown to be due to a single-nucleotide polymorphism located between the adult alpha-globin genes and their enhancers. The finding that this mutation creates a novel promoter provides support for a mechanism of gene regulation by facilitated chromatin looping. PMID- 17076880 TI - Getting a buzz out of the bee genome. AB - The honey bee Apis mellifera displays the most complex behavior of any insect. This, and its utility to humans, makes it a fascinating object of study for biologists. Such studies are now further enabled by the release of the honey-bee genome sequence. PMID- 17076881 TI - Long-term consequences of arsenic poisoning during infancy due to contaminated milk powder. AB - Arsenic toxicity is a global health problem affecting many millions of people. The main source of exposure is drinking water contaminated by natural geological sources. Current risk assessment is based on the recognized carcinogenicity of arsenic, but neurotoxic risks have been overlooked. In 1955, an outbreak of arsenic poisoning occurred among Japanese infants, with more than 100 deaths. The source was contaminated milk powder produced by the Morinaga company. Detailed accounts of the Morinaga dried milk poisoning were published in Japanese only, and an overview of this poisoning incident and its long-term consequences is therefore presented. From analyses available, the arsenic concentration in milk made from the Morinaga milk powder is calculated to be about 4-7 mg/L, corresponding to daily doses slightly above 500 microg/kg body weight. Lower exposures would result from using diluted milk. Clinical poisoning cases occurred after a few weeks of exposure, with a total dose of about 60 mg. This experience provides clear-cut evidence for hazard assessment of the developmental neurotoxicity. At the present time, more than 600 surviving victims, now in their 50s, have been reported to suffer from severe sequelae, such as mental retardation, neurological diseases, and other disabilities. Along with more recent epidemiological studies of children with environmental arsenic exposures, the data amply demonstrate the need to consider neurotoxicity as a key concern in risk assessment of inorganic arsenic exposure. PMID- 17076882 TI - Serum neopterin levels in patients with replicative and nonreplicative HBV carriers. AB - BACKGROUND: Infection by hepatitis B virus (HBV) causes complicated biochemical, immunological and histological changes in host immune response against the virus which can be specific or non-specific. Recent attention has focused on neopterin as a marker for the activation of cell mediated immunity. The aim of this study was to define the pattern of neopterin levels in replicative and nonreplicative HBV carriers. METHODS: Thirty HBV replicative carriers and 25 nonreplicative HBV carriers and 30 healthy adult patients were included this study. Hepatitis markers were determined by commercial kit based on chemilumminesans assay. HBV DNA was quantified by hybrid capture system. Serum neopterin levels were measured by the method of competitive enzyme-linked immunosorbent assay. Results were expressed as mean +/- SD and ranges. RESULTS: In the nonreplicative group, except for one patient, all the patients' HBeAg were negative and anti-HBe were positive. That particular patient was HBeAg positive and anti-HBe negative. In the replicative group, 23 out of 30 patients have positive HBeAg and negative anti-HBe; 7 out of 30 patients have negative HBeAg and positive anti-HBe. Serum neopterin concentrations were 14.5 +/- 10.0 (4.2-41) nmol/L in replicative HBV carriers, 8.9 +/- 4.3 (2.1-22) nmol/L in nonreplicative HBV carriers and 7.1 +/- 2.2 (4.0-12) nmol/L in the control group. Serum neopterin levels and the rates of abnormal serum neopterin levels in the replicative group were higher than the control group (P < 0.01 and P < 0.05). In the nonreplicative group, serum neopterin levels were not different from those of the control. There was a difference between replicative and nonreplicative groups in the respect of neopterin levels. CONCLUSION: In the hepatitis B infected carriers, elevated neopterin levels may be an indicator of the presence of replication. PMID- 17076883 TI - Coping strategies, stress, physical activity and sleep in patients with unexplained chest pain. AB - BACKGROUND: The number of patients suffering from unexplained chest pain (UCP) is increasing. Intervention programmes are needed to reduce the chest pain and suffering experienced by these patients and effective preventive strategies are also required to reduce the incidence of these symptoms. The aim of this study was to describe general coping strategies in patients with UCP and examine the relationships between coping strategies, negative life events, sleep problems, physical activity, stress and chest pain intensity. METHOD: The sample consisted of 179 patients younger than 70 years of age, who were evaluated for chest pain at the emergency department daytime Monday through Friday and judged by a physician to have no organic cause for their chest pain. The study had a cross sectional design. RESULTS: Emotive coping was related to chest pain intensity (r = 0.17, p = 0.02). Women used emotive coping to a greater extent than did men (p = 0.05). In the multivariate analysis was shown that physical activity decreased emotive coping (OR 0.13, p < 0.0001) while sex, age, sleep, mental strain at work and negative life events increased emotive coping. Twenty-seven percent of the patients had sleep problems 8 to 14 nights per month or more. Permanent stress at work during the last year was reported by 18% of the patients and stress at home by 7%. Thirty-five percent of the patients were worried often or almost all the time about being rushed at work and 23% were worried about being unable to keep up with their workload. Concerning total life events, 20% reported that a close relative had had a serious illness and 27% had reasons to be worried about a close relative. CONCLUSION: Our results indicated that patients with more intense UCP more often apply emotive coping in dealing with their pain. Given that emotive coping was also found to be related to disturbed sleep, negative life events, mental strain at work and physical activity, it may be of value to help these patients to both verbalise their emotions and to become cognizant of the influence of such factors on their pain experience. PMID- 17076884 TI - Parents' knowledge and behaviour concerning sunning their babies; a cross sectional, descriptive study. AB - BACKGROUND: For centuries, sunlight has been used for therapeutic purposes. Parents still sun their infants to treat neonatal jaundice, nappy rash or mostly to supply vitamin D for bone development as a consequence of health beliefs. In this study we aimed to assess knowledge and behaviour of parents about benefits of sunlight and sun protection. METHODS: In this study, parents attending to governmental primary healthcare units for their children's routine vaccinations, upon their informed consent, were consecutively enrolled during one month. Data were collected by a semi-structured questionnaire. RESULTS: The mean age of 118 enrolled parents and their babies were 27.9 +/- 6.5 years and 8.3 +/- 5.8 months, respectively. Most of the participants were mothers (93.2%), housewives (81.4%) with an educational level of > or =6 years (71.2%). Sunlight was considered beneficial for bone development (86.4%), diaper rash (5.9%) and neonatal jaundice (12.7%). In case of neonatal jaundice 72.0% of the participants reported that they would consult a physician. Most of the participants (82.2%) were sunning their babies outdoors. Nearly half (49.7%) of them got this information from medical staff. Fifty two percent of the parents were sunning their babies before 10-11 a.m. and/or after 3 p.m. Only 13.6% of parents reported using sunscreen for their babies, and the majority of them were using sun protecting factor > or = 15. One forth of the sunscreen users was using sunscreen according to their physicians' advice. CONCLUSION: Most of the participants were aware of the benefits of sunlight; especially for bone development. However they were displaying inappropriate behaviour while sunning their babies for health reasons. More education should be given to parents about the danger of sunlight at primary health care units while advising to sun their babies, if any. PMID- 17076885 TI - Evaluation of the accuracy of serum MMP-9 as a test for colorectal cancer in a primary care population. AB - BACKGROUND: Bowel cancer is common and is a major cause of death. Meta-analysis of randomised controlled trials estimates that screening for colorectal cancer using faecal occult blood (FOB) test reduces mortality from colorectal cancer by 16%. However, FOB testing has a low positive predictive value, with associated unnecessary cost, risk and anxiety from subsequent investigation, and is unacceptable to a proportion of the target population. Increased levels of an enzyme called matrix metalloproteinase 9 (MMP-9) have been found to be associated with colorectal cancer, and this can be measured from a blood sample. Serum MMP-9 is potentially an accurate, low risk and cost-effective population screening tool. This study aims to evaluate the accuracy of serum MMP-9 as a test for colorectal cancer in a primary care population. METHODS/DESIGN: People aged 50 to 69 years, who registered in participating general practices in the West Midlands Region, will be asked to complete a questionnaire that asks about symptoms. Respondents who describe any colorectal symptoms (except only abdominal bloating and/or anal symptoms) and are prepared to provide a blood sample for MMP9 estimation and undergo a colonoscopy (current gold standard investigation) will be recruited at GP based clinics by a research nurse. Those unfit for colonoscopy will be excluded. Colonoscopies will be undertaken in dedicated research clinics. The accuracy of MMP-9 will be assessed by comparing the MMP-9 level with the colonoscopy findings, and the combination of factors (e.g. symptoms and MMP-9 level) that best predict a diagnosis of malignancy (invasive disease or polyps) will be determined. DISCUSSION: Colorectal cancer is a major cause of morbidity and mortality. Most colorectal cancers arise from adenomas and there is a period for early detection by screening, but available tests have risks, are unacceptable to many, have high false positive rates or are expensive. This study will establish the potential of serum MMP-9 as a screening test for colorectal cancer. If it is confirmed as accurate and acceptable, this serum marker has the potential to assist with reducing the morbidity and mortality from colorectal cancer. PMID- 17076886 TI - Eating and weight related cognitions in people with Schizophrenia : a case control study. AB - BACKGROUND: Patients with antipsychotic-induced weight gain (WG) regularly report on unsuccessful dietary trials, which suggests strong biological weight gain drive that is extremely hard to overcome with thoughts, such that behaviour doesn't change despite some intent to change. The purpose of the present study was to assess cognitions specifically related to restrained eating in severely overweight patients with schizophrenia treated with antipsychotic drugs. METHODS: Forty outpatients with schizophrenia and 40 controls without psychiatric disability were included. Both groups were composed of one subgroup severely overweight (defined as a BMI > 28), and a comparison sample (BMI<28). The revised version of the Mizes Anorectic cognitive questionnaire (MAC-R) was used in this cross-sectional case-control study. RESULTS: Gender was significantly related to eating disorders cognition, women scoring higher than men. Patients with schizophrenia in general scored higher on the MAC-R total scale and on the MAC-R subscale 2, the latter score representing rigid weight regulation and fear of weight gain. When comparing the two groups of subjects with BMI < 28, it appeared that patients with schizophrenia also scored higher on MAC-R total scale, the subscales 2 and 3, the latter subscale 3, indicating altered self control and self-esteem. CONCLUSION: As is the case in weight gain of subjects without schizophrenia, the present results suggest that the cognitive distortions, as assessed by the MAC-R, may play an important role in weight gain also in patients with schizophrenia, and in weight gain associated with antipsychotic pharmacotherapy. Particular attention to these processes may help to improve the management of antipsychotic drugs induced weight gain. PMID- 17076887 TI - RiboSubstrates: a web application addressing the cleavage specificities of ribozymes in designated genomes. AB - BACKGROUND: RNA-dependent gene silencing is becoming a routine tool used in laboratories worldwide. One of the important remaining hurdles in the selection of the target sequence, if not the most important one, is the designing of tools that have minimal off-target effects (i.e. cleaves only the desired sequence). Increasingly, in the current dawn of the post-genomic era, there is a heavy reliance on tools that are suitable for high-throughput functional genomics, consequently more and more bioinformatic software is becoming available. However, to date none have been designed to satisfy the ever-increasing need for the accurate selection of targets for a specific silencing reagent. RESULTS: In order to overcome this hurdle we have developed RiboSubstrates http://www.riboclub.org/ribosubstrates. This integrated bioinformatic software permits the searching of a cDNA database for all potential substrates for a given ribozyme. This includes the mRNAs that perfectly match the specific requirements of a given ribozyme, as well those including Wobble base pairs and mismatches. The results generated allow rapid selection of sequences suitable as targets for RNA degradation. The current web-based RiboSubstrates version permits the identification of potential gene targets for both SOFA-HDV ribozymes and for hammerhead ribozymes. Moreover, a minimal template for the search of siRNAs is also available. This flexible and reliable tool is easily adaptable for use with any RNA tool (i.e. other ribozymes, deoxyribozymes and antisense), and may use the information present in any cDNA bank. CONCLUSION: RiboSubstrates should become an essential step for all, even including "non-RNA biologists", who endeavor to develop a gene-inactivation system. PMID- 17076888 TI - Accuracy of spinal orthopaedic tests: a systematic review. AB - BACKGROUND: The purpose of this systematic review was to critically appraise the literature on the accuracy of orthopaedic tests for the spine. METHODS: Multiple orthopaedic texts were reviewed to produce a comprehensive list of spine orthopaedic test names and synonyms. A search was conducted in MEDLINE, MANTIS, CINAHL, AMED and the Cochrane Library for relevant articles from inception up to December 2005. The studies were evaluated using the tool for quality assessment for diagnostic accuracy studies (QUADAS). RESULTS: Twenty-one papers met the inclusion criteria. The QUADAS scores ranged from 4 to 12 of a possible 14. Twenty-nine percent of the studies achieved a score of 10 or more. The papers covered a wide range of tests for spine conditions. CONCLUSION: There was a lack of quantity and quality of orthopaedic tests for the spine found in the literature. There is a lack of high quality research regarding the accuracy of spinal orthopaedic tests. Due to this lack of evidence it is suggested that over reliance on single orthopaedic tests is not appropriate. PMID- 17076889 TI - Comprehensive analysis of gene expression patterns of hedgehog-related genes. AB - BACKGROUND: The Caenorhabditis elegans genome encodes ten proteins that share sequence similarity with the Hedgehog signaling molecule through their C-terminal autoprocessing Hint/Hog domain. These proteins contain novel N-terminal domains, and C. elegans encodes dozens of additional proteins containing only these N terminal domains. These gene families are called warthog, groundhog, ground-like and quahog, collectively called hedgehog (hh)-related genes. Previously, the expression pattern of seventeen genes was examined, which showed that they are primarily expressed in the ectoderm. RESULTS: With the completion of the C. elegans genome sequence in November 2002, we reexamined and identified 61 hh related ORFs. Further, we identified 49 hh-related ORFs in C. briggsae. ORF analysis revealed that 30% of the genes still had errors in their predictions and we improved these predictions here. We performed a comprehensive expression analysis using GFP fusions of the putative intergenic regulatory sequence with one or two transgenic lines for most genes. The hh-related genes are expressed in one or a few of the following tissues: hypodermis, seam cells, excretory duct and pore cells, vulval epithelial cells, rectal epithelial cells, pharyngeal muscle or marginal cells, arcade cells, support cells of sensory organs, and neuronal cells. Using time-lapse recordings, we discovered that some hh-related genes are expressed in a cyclical fashion in phase with molting during larval development. We also generated several translational GFP fusions, but they did not show any subcellular localization. In addition, we also studied the expression patterns of two genes with similarity to Drosophila frizzled, T23D8.1 and F27E11.3A, and the ortholog of the Drosophila gene dally-like, gpn-1, which is a heparan sulfate proteoglycan. The two frizzled homologs are expressed in a few neurons in the head, and gpn-1 is expressed in the pharynx. Finally, we compare the efficacy of our GFP expression effort with EST, OST and SAGE data. CONCLUSION: No bona-fide Hh signaling pathway is present in C. elegans. Given that the hh-related gene products have a predicted signal peptide for secretion, it is possible that they constitute components of the extracellular matrix (ECM). They might be associated with the cuticle or be present in soluble form in the body cavity. They might interact with the Patched or the Patched-related proteins in a manner similar to the interaction of Hedgehog with its receptor Patched. PMID- 17076890 TI - Comparison of 18F SPECT with PET in myocardial imaging: a realistic thorax cardiac phantom study. AB - BACKGROUND: Positron emission tomography (PET) imaging with fluorine-18 (18F) Fluorodeoxyglucose (FDG) and flow tracer such as Rubidium-82 (82Rb) is an established method for evaluating an ischemic but viable myocardium. However, the high cost of PET imaging restricts its wider clinical use. Therefore, less expensive 18F FDG single photon emission computed tomography (SPECT) imaging has been considered as an alternative to 18F FDG PET imaging. The purpose of the work is to compare SPECT with PET in myocardial perfusion/viability imaging. METHODS: A nonuniform RH-2 thorax-heart phantom was used in the SPECT and PET acquisitions. Three inserts, 3 cm, 2 cm and 1 cm in diameter, were placed in the left ventricular (LV) wall to simulate infarcts. The phantom acquisition was performed sequentially with 7.4 MBq of 18F and 22.2 MBq of Technetium-99m (99mTc) in the SPECT study and with 7.4 MBq of 18F and 370 MBq of 82Rb in the PET study. SPECT and PET data were processed using standard reconstruction software provided by vendors. Circumferential profiles of the short-axis slices, the contrast and viability of the inserts were used to evaluate the SPECT and PET images. RESULTS: The contrast for 3 cm, 2 cm and 1 cm inserts were for 18F PET data, 1.0 +/- 0.01, 0.67 +/- 0.02 and 0.25 +/- 0.01, respectively. For 82Rb PET data, the corresponding contrast values were 0.61 +/- 0.02, 0.37 +/- 0.02 and 0.19 +/- 0.01, respectively. For 18F SPECT the contrast values were, 0.31 +/- 0.03 and 0.20 +/- 0.05 for 3 cm and 2 cm inserts, respectively. For 99mTc SPECT the contrast values were, 0.63 +/- 0.04 and 0.24 +/- 0.05 for 3 cm and 2 cm inserts respectively. In SPECT, the 1 cm insert was not detectable. In the SPECT study, all three inserts were falsely diagnosed as "viable", while in the PET study, only the 1 cm insert was diagnosed falsely "viable". CONCLUSION: For smaller defects the 99mTc/18F SPECT imaging cannot entirely replace the more expensive 82Rb/18F PET for myocardial perfusion/viability imaging, due to poorer image spatial resolution and poorer defect contrast. PMID- 17076891 TI - The FDA guidance for industry on PROs: the point of view of a pharmaceutical company. AB - The importance of the patients point of view on their health status is widely recognised. Patient-reported outcomes is a broad term encompassing a large variety of different health data reported by patients, as symptoms, functional status, Quality of Life and Health-Related Quality of Life. Measurements of Health-Related Quality of Life have been developed during many years of researches, and a lot of validated questionnaires exist. However, few attempts have been made to standardise the evaluation of instruments characteristics, no recommendations are made about interpretation on Health-Related Quality of Life results, especially regarding the clinical significance of a change leading a therapeutic approach. Moreover, the true value of Health-Related Quality of Life evaluations in clinical trials has not yet been completely defined. An important step towards a more structured and frequent use of Patient-Reported Outcomes in drug development is represented by the FDA Guidance, issued on February 2006. In our paper we aim to report some considerations on this Guidance. Our comments focus especially on the characteristics of instruments to use, the Minimal Important Difference, and the methods to calculate it. Furthermore, we present the advantages and opportunities of using the Patient-Reported Outcomes in drug development, as seen by a pharmaceutical company. The Patient-Reported Outcomes can provide additional data to make a drug more competitive than others of the same pharmacological class, and a well demonstrated positive impact on the patient' health status and daily life might allow a higher price and/or the inclusion in a reimbursement list. Applying extensively the FDA Guidance in the next trials could lead to a wider culture of subjective measurement, and to a greater consideration for the patient's opinions on his/her care. Moreover, prescribing doctors and payers could benefit from subjective information to better define the value of drugs. PMID- 17076892 TI - Transition of healthy to diseased synovial tissue in rheumatoid arthritis is associated with gain of mesenchymal/fibrotic characteristics. AB - The healthy synovial lining layer consists of a single cell layer that regulates the transport between the joint cavity and the surrounding tissue. It has been suggested that abnormalities such as somatic mutations in the p53 tumor suppressor gene contribute to synovial hyperplasia and invasion in rheumatoid arthritis (RA). In this study, expression of epithelial markers on healthy and diseased synovial lining tissue was examined. In addition, we investigated whether a regulated process, resembling epithelial to mesenchymal transition (EMT)/fibrosis, could be responsible for the altered phenotype of the synovial lining layer in RA. Synovial tissue from healthy subjects and RA patients was obtained during arthroscopy. To detect signs of EMT, expression of E-cadherin (epithelial marker), collagen type IV (indicator of the presence of a basement membrane) and alpha-smooth muscle actin (alpha-sma; a myofibroblast marker) was investigated on frozen tissue sections using immunohistochemistry. Fibroblast like synoviocytes (FLSs) from healthy subjects were isolated and subjected to stimulation with synovial fluid (SF) from two RA patients and to transforming growth factor (TGF)-beta. To detect whether EMT/fibrotic markers were increased, expression of collagen type I, alpha-sma and telopeptide lysylhydroxylase (TLH) was measured by real time PCR. Expression of E-cadherin and collagen type IV was found in healthy and arthritic synovial tissue. Expression of alpha-sma was only found in the synovial lining layer of RA patients. Stimulation of healthy FLSs with SF resulted in an upregulation of alpha-sma and TLH mRNA. Collagen type I and TLH mRNA were upregulated after stimulation with TGF-beta. Addition of bone morphogenetic protein (BMP)-7 to healthy FLS stimulated with SF inhibited the expression of alpha-sma mRNA. The finding that E-cadherin and collagen type IV are expressed in the lining layer of healthy and arthritic synovium indicates that these lining cells display an epithelial-like phenotype. In addition, the presence of alpha-sma in the synovial lining layer of RA patients and induction of fibrotic markers in healthy FLSs by SF from RA patients indicate that a regulated process comparable to EMT might cause the alteration in phenotype of RA FLSs. Therefore, BMP-7 may represent a promising agent to counteract the transition imposed on synoviocytes in the RA joint. PMID- 17076893 TI - Radiological, pathological and DNA remission in recurrent metastatic nasopharyngeal carcinoma. AB - BACKGROUND: Circulating plasma Epstein Barr Virus DNA (EBV-DNA) is a sensitive and specific marker of nasopharyngeal carcinoma (NPC). The mainstay of treatment of metastatic NPC is systemic chemotherapy and resection for solitary metastasis. Despite high response rate to chemotherapy, complete remission is uncommonly seen. CASE PRESENTATION: We report a case of recurrent metastatic NPC in a 43 year-old man, who achieved complete remission three times with chemotherapy and surgery. Serial plasma EBV-DNA levels were measured during the course of disease. The patient had three episodes of recurrences of NPC manifested as distant metastasis. Both time, rise in the plasma EBV-DNA level preceded detection of recurrences by imaging. Following systemic chemotherapy, he achieved complete remission each time, of which was confirmed by 18-flourodeoxyglucose positron emission tomography and hepatectomy pathology. The plasma EBV-DNA level dropped to zero copy/ml at the time of each remission. CONCLUSION: This case highlights the high chemosensitivity of NPC by illustrating a rare occurrence of complete response of metastatic NPC to chemotherapy. This case also underscores the usefulness of EBV-DNA as a useful tool in monitoring NPC by its ability to detect early recurrence and excellent correlation with treatment response. PMID- 17076894 TI - Transformation of Anaplasma phagocytophilum. AB - BACKGROUND: Tick-borne pathogens cause emerging zoonoses, and include fastidious organisms such as Anaplasma phagocytophilum. Because of their obligate intracellular nature, methods for mutagenesis and transformation have not been available. RESULTS: To facilitate genetic manipulation, we transformed A. phagocytophilum (Ap) to express a green fluorescent protein (GFP) with the Himar1 transposase system and selection with the clinically irrelevant antibiotic spectinomycin. CONCLUSION: These transformed bacteria (GFP/Ap) grow at normal rates and are brightly fluorescent in human, monkey, and tick cell culture. Molecular characterization of the GFP/Ap genomic DNA confirmed transposition and the flanking genomic insertion locations were sequenced. Three mice inoculated with GFP/Ap by intraperitoneal injection became infected as demonstrated by the appearance of morulae in a peripheral blood neutrophil and re-isolation of the bacteria in culture. PMID- 17076895 TI - CellProfiler: image analysis software for identifying and quantifying cell phenotypes. AB - Biologists can now prepare and image thousands of samples per day using automation, enabling chemical screens and functional genomics (for example, using RNA interference). Here we describe the first free, open-source system designed for flexible, high-throughput cell image analysis, CellProfiler. CellProfiler can address a variety of biological questions quantitatively, including standard assays (for example, cell count, size, per-cell protein levels) and complex morphological assays (for example, cell/organelle shape or subcellular patterns of DNA or protein staining). PMID- 17076896 TI - History and development of trauma registry: lessons from developed to developing countries. AB - BACKGROUND: A trauma registry is an integral component of modern comprehensive trauma care systems. Trauma registries have not been established in most developing countries, and where they exist are often rudimentary and incomplete. This review describes the role of trauma registries in the care of the injured, and discusses how lessons from developed countries can be applied toward their design and implementation in developing countries. METHODS: A detailed review of English-language articles on trauma registry was performed using MEDLINE and CINAHL. In addition, relevant articles from non-indexed journals were identified with Google Scholar. RESULTS: The history and development of trauma registries and their role in modern trauma care are discussed. Drawing from past and current experience, guidelines for the design and implementation of trauma registries are given, with emphasis on technical and logistic factors peculiar to developing countries. CONCLUSION: Improvement in trauma care depends on the establishment of functioning trauma care systems, of which a trauma registry is a crucial component. Hospitals and governments in developing countries should be encouraged to establish trauma registries using proven cost-effective strategies. PMID- 17076898 TI - ADDP and PS-PPh3: an efficient Mitsunobu protocol for the preparation of pyridine ether PPAR agonists. AB - A series of pyridine ether PPAR agonists were synthesized through an ADDP and PS PPh3 modified Mitsunobu protocol, which eliminated significant by-product formation. This method proved to be versatile, efficient and amenable to parallel synthesis. PMID- 17076897 TI - A consensus prognostic gene expression classifier for ER positive breast cancer. AB - BACKGROUND: A consensus prognostic gene expression classifier is still elusive in heterogeneous diseases such as breast cancer. RESULTS: Here we perform a combined analysis of three major breast cancer microarray data sets to hone in on a universally valid prognostic molecular classifier in estrogen receptor (ER) positive tumors. Using a recently developed robust measure of prognostic separation, we further validate the prognostic classifier in three external independent cohorts, confirming the validity of our molecular classifier in a total of 877 ER positive samples. Furthermore, we find that molecular classifiers may not outperform classical prognostic indices but that they can be used in hybrid molecular-pathological classification schemes to improve prognostic separation. CONCLUSION: The prognostic molecular classifier presented here is the first to be valid in over 877 ER positive breast cancer samples and across three different microarray platforms. Larger multi-institutional studies will be needed to fully determine the added prognostic value of molecular classifiers when combined with standard prognostic factors. PMID- 17076899 TI - Drug resistance to sulphadoxine-pyrimethamine in Plasmodium falciparum malaria in Mlimba, Tanzania. AB - BACKGROUND: Sulphadoxine-pyrimethamine (SP) has been and is currently used for treatment of uncomplicated Plasmodium falciparum malaria in many African countries. Nevertheless, the response of parasites to SP treatment has shown significant variation between individuals. METHODS: The genes for dihydrofolate reductase (dhfr) and dihydropteroate synthase (dhps) were used as markers, to investigate parasite resistance to SP in 141 children aged less than 5 years. Parasite DNA was extracted by Chelex method from blood samples collected and preserved on filter papers. Subsequently, polymerase chain reaction (PCR) and restriction fragment length polymorphism (PCR-RFLP) were applied to detect the SP resistance-associated point mutations on dhfr and dhps. Commonly reported point mutations at codons 51, 59, 108 and 164 in the dhfr and codons 437, 540 and 581 in the dhps domains were examined. RESULTS: Children infected with parasites harbouring a range of single to quintuple dhfr/dhps mutations were erratically cured with SP. However, the quintuple dhfr/dhps mutant genotypes were mostly associated with treatment failures. High proportion of SP resistance-associated point mutations was detected in this study but the adequate clinical response (89.4%) observed clinically at day 14 of follow up reflects the role of semi immunity protection and parasite clearance in the population. CONCLUSION: In monitoring drug resistance to SP, concurrent studies on possible confounding factors pertaining to development of resistance in falciparum malaria should be considered. The SP resistance potential detected in this study, cautions on its useful therapeutic life as an interim first-line drug against malaria in Tanzania and other malaria-endemic countries. PMID- 17076900 TI - An improved and highly sensitive microfluorimetric method for assessing susceptibility of Plasmodium falciparum to antimalarial drugs in vitro. AB - BACKGROUND: The standard in vitro protocol currently in use for drug testing against Plasmodium falciparum, based on the incorporation of the purine [3H] hypoxanthine, has two serious drawbacks. Firstly it is unsuitable for the testing of drugs that directly or indirectly impact on purine salvage or metabolism. Secondly, it relies on the use of expensive radiolabelled material, with added issues concerning detection, storage and waste disposal that make it unsuitable for use in many disease-endemic areas. Recently, the use of fluorochromes has been suggested as an alternative, but quenching of the fluorescence signal by the haemoglobin present in cultures of Plasmodium falciparum-infected erythrocytes severely limits the usefulness of this approach. METHODS: In order to resolve this problem, a new PicoGreen-based procedure has been developed which incorporates additional steps to remove the interfering haemoglobin. The 50% inhibitory concentration (IC50) values of chloroquine and pyrimethamine against P. falciparum laboratory lines 3D7 and K1 were determined using the new protocol. RESULTS: The IC50 values of chloroquine and pyrimethamine against P. falciparum laboratory lines 3D7 and K1 determined with the new fluorescence-based protocol were statistically identical to those obtained using the traditional 3H hypoxanthine incorporation method, and consistent with literature values. CONCLUSION: The new method proved to be accurate, reproducible and sensitive, and has the advantage of being non-radioactive. The improved PicoGreen method has the potential to replace traditional in vitro drug resistance assay techniques. PMID- 17076901 TI - Deriving utility scores for co-morbid conditions: a test of the multiplicative model for combining individual condition scores. AB - BACKGROUND: The co-morbidity of health conditions is becoming a significant health issue, particularly as populations age, and presents important methodological challenges for population health research. For example, the calculation of summary measures of population health (SMPH) can be compromised if co-morbidity is not taken into account. One popular co-morbidity adjustment used in SMPH computations relies on a straightforward multiplicative combination of the severity weights for the individual conditions involved. While the convenience and simplicity of the multiplicative model are attractive, its appropriateness has yet to be formally tested. The primary objective of the current study was therefore to examine the empirical evidence in support of this approach. METHODS: The present study drew on information on the prevalence of chronic conditions and a utility-based measure of health-related quality of life (HRQoL), namely the Health Utilities Index Mark 3 (HUI3), available from Cycle 1.1 of the Canadian Community Health Survey (CCHS; 2000-01). Average HUI3 scores were computed for both single and co-morbid conditions, and were also purified by statistically removing the loss of functional health due to health problems other than the chronic conditions reported. The co-morbidity rule was specified as a multiplicative combination of the purified average observed HUI3 utility scores for the individual conditions involved, with the addition of a synergy coefficient s for capturing any interaction between the conditions not explained by the product of their utilities. The fit of the model to the purified average observed utilities for the co-morbid conditions was optimized using ordinary least squares regression to estimate s. Replicability of the results was assessed by applying the method to triple co-morbidities from the CCHS cycle 1.1 database, as well as to double and triple co-morbidities from cycle 2.1 of the CCHS (2003 04). RESULTS: Model fit was optimized at s = .99 (i.e., essentially a straightforward multiplicative model). These results were closely replicated with triple co-morbidities reported on CCHS 2000-01, as well as with double and triple co-morbidities reported on CCHS 2003-04. CONCLUSION: The findings support the simple multiplicative model for computing utilities for co-morbid conditions from the utilities for the individual conditions involved. Future work using a wider variety of conditions and data sources could serve to further evaluate and refine the approach. PMID- 17076902 TI - Utilizing FEM-Software to quantify pre- and post-interventional cardiac reconstruction data based on modelling data sets from surgical ventricular repair therapy (SVRT) and cardiac resynchronisation therapy (CRT). AB - BACKGROUND: Left ventricle (LV) 3D structural data can be easily obtained using standard transesophageal echocardiography (TEE) devices but quantitative pre- and intraoperative volumetry and geometry analysis of the LV is presently not feasible in the cardiac operation room (OR). Finite element method (FEM) modelling is necessary to carry out precise and individual volume analysis and in the future will form the basis for simulation of cardiac interventions. METHOD: A Philips/HP Sonos 5500 ultrasound device stores volume data as time-resolved 4D volume data sets. In this prospective study TomTec LV Analysis TEE Software was used for semi-automatic endocardial border detection, reconstruction, and volume rendering of the clinical 3D echocardiographic data. With the software FemCoGen a quantification of partial volumes and surface directions of the LV was carried out for two patients data sets. One patient underwent surgical ventricular repair therapy (SVR) and the other a cardiac resynchronisation therapy (CRT). RESULTS: For both patients a detailed volume and surface direction analysis is provided. Partial volumes as well as normal directions to the LV surface are pre- and post interventionally compared. CONCLUSION: The operation results for both patients are quantified. The quantification shows treatment details for both interventions (e.g. the elimination of the discontinuities for CRT intervention and the segments treated for SVR intervention). The LV quantification is feasible in the cardiac OR and it gives a detailed and immediate quantitative feedback of the quality of the intervention to the medical. PMID- 17076903 TI - Occurrence of haemolytic Mannheimia spp. in apparently healthy sheep in Norway. AB - BACKGROUND: The occurrence of Mannheimia species in healthy sheep has only been investigated to a very limited extend since the genus and its five named species were established. The aim of the present study was to evaluate the occurrence of haemolytic Mannheimia species in apparently healthy sheep originating from four sheep flocks in South-Western Norway. METHODS: Typical beta-haemolytic Pasteurellaceae were isolated from nasal swabs and subsequently subjected to bacteriological examination. A total of 57 Mannheimia isolates were obtained in pure culture. All isolates were genotyped by amplified fragment length polymorphisms (AFLP) analysis and compared to six reference strains. The 16S rRNA gene sequences of two isolates were also determined. RESULTS: beta-haemolytic Mannheimia species were isolated from 24% to 64% of the sheep in the four flocks. A total of 26 haemolytic M. ruminalis-like strains were isolated among which, a considerable genetic diversity was found. Eighteen M. glucosida isolates were obtained from three flocks, whereas M. haemolytica was only isolated from two flocks, 16 of them being from only one of the flocks. CONCLUSION: We demonstrate that a relatively high number of apparently healthy sheep in Norway seem to carry the potentially pathogenic M. haemolytica and M. glucosida in the upper respiratory tract. An unexpectedly high number of haemolytic M. ruminalis-like organisms were also obtained in all four flocks. The usually non-haemolytic M. ruminalis are typically isolated from healthy ruminants. The significance of beta haemolytic M. ruminalis-like organisms is unknown and should be investigated in a future study. PMID- 17076904 TI - Dietary patterns and associated lifestyles in individuals with and without familial history of obesity: a cross-sectional study. AB - BACKGROUND: Familial history of obesity (FHO) and certain dietary habits are risk factors for obesity. The objectives of this cross-sectional study were 1) to derive dietary patterns using factor analysis in a population of men and women with and without FHO; 2) to compare mean factor scores for each dietary pattern between individuals with and without FHO; and 3) to examine the association between these patterns and anthropometric, lifestyle and sociodemographic variables. METHODS: A total of 197 women and 129 men with a body mass index <30 kg/m2 were recruited. A positive FHO (FHO+) was defined as having at least one obese first-degree relative and a negative FHO (FHO-) as no obese first-degree relative. Dietary data were collected from a food frequency questionnaire. Factor analysis was performed to derive dietary patterns. Mean factor scores were compared using general linear model among men and women according to FHO. Regression analyses were performed to study the relationship between anthropometric, lifestyle and sociodemographic variables, and each dietary pattern. RESULTS: Two dietary patterns were identified in both men and women : the Western pattern characterized by a higher consumption of red meats, poultry, processed meats, refined grains as well as desserts, and the Prudent pattern characterized by greater intakes of vegetables, fruits, non-hydrogenated fat, and fish and seafood. Similar Western and Prudent factor scores were observed in individual with and without FHO. In men with FHO+, the Western pattern is negatively associated with age and positively associated with physical activity, smoking, and personal income. In women with FHO-, the Prudent pattern is negatively associated with BMI and smoking and these pattern is positively associated with age and physical activity. CONCLUSION: Two dietary patterns have been identified among men and women with and without FHO. Although that FHO does not seem to influence the adherence to dietary patterns, results of this study suggest that anthropometric, lifestyle and sociodemographic variables associated with dietary patterns differ according to FHO and gender. PMID- 17076905 TI - Immunogenicity of a polyvalent HIV-1 candidate vaccine based on fourteen wild type gp120 proteins in golden hamsters. AB - BACKGROUND: One of the major obstacles in the design of an effective vaccine against HIV-1 is the hypervariability of the HIV-1 envelope glycoprotein. Most HIV-1 vaccine candidates have utilized envelope glycoprotein from a single virus isolate, but to date, none of them elicited broadly reactive humoral immunity. Herein, we hypothesised that a cocktail of HIV-1 gp120 proteins containing multiple epitopes may increase the breadth of immune responses against HIV-1. We compared and evaluated the immunogenicity of HIV-1 vaccines containing either gp120 protein alone or in combinations of four or fourteen gp120s from different primary HIV-1 isolates in immunized hamsters. RESULTS: We amplified and characterized 14 different gp120s from primary subtype B isolates with both syncytium and non-syncytium inducing properties, and expressed the proteins in Chinese Hamster Ovary (CHO) cell lines. Purified proteins were used either alone or in combinations of four or fourteen different gp120s to vaccinate golden hamsters. The polyvalent vaccine showed higher antibody titers to HIV-1 subtype B isolates MN and SF162 compared to the groups that received one or four gp120 proteins. However, the polyvalent vaccine was not able to show higher neutralizing antibody responses against HIV-1 primary isolates. Interestingly, the polyvalent vaccine group had the highest proliferative immune responses and showed a substantial proportion of cross-subtype CD4 reactivity to HIV-1 subtypes B, C, and A/E CONCLUSION: Although the polyvalent approach achieved only a modest increase in the breadth of humoral and cellular immunity, the qualitative change in the vaccine (14 vs. 1 gp120) resulted in a quantitative improvement in vaccine induced immunity. PMID- 17076906 TI - The influence of partnership centrality on organizational perceptions of support: a case study of the AHLN structure. AB - BACKGROUND: Knowledge of the structure and character of inter-organizational relationships found among health promotion organizations is a prerequisite for the development of evidence-based network-level intervention activities. The Alberta Healthy Living Network (AHLN) mapped the inter-organizational structure of its members to examine the effects of the network environment on organizational-level perceptions. This exploratory analysis examines whether network structure, specifically partnership ties among AHLN members, influences organizational perceptions of support after controlling for organizational-level attributes. METHODS: Organizational surveys were conducted with representatives from AHLN organizations as of February 2004 (n = 54). Organizational attribute and inter-organizational data on various network dimensions were collected. Organizations were classified into traditional and non-traditional categories. We examined the partnership network dimension. In- and out-degree centrality scores on partnership ties were calculated for each organization and tested against organizational perceptions of available financial support. RESULTS: Non traditional organizations are more likely to view financial support as more readily available for their HEALTR programs and activities than traditional organizations (1.57, 95% CI: .34, 2.79). After controlling for organizational characteristics, organizations that have been frequently identified by other organizations as valuable partners in the AHLN network were found significantly more likely to perceive a higher sense of funding availability (In-degree partnership value) (.03, 95% CI: .01, .05). CONCLUSION: Organizational perceptions of a supportive environment are framed not only by organizational characteristics but also by an organization's position in an inter-organizational network. Network contexts can influence the way that organizations perceive their environment and potentially the actions that organizations may take in light of such perceptions. By developing evidence-based understandings on the influence of network contexts, the AHLN can better target the particularities of its specific health promotion network. PMID- 17076907 TI - Sequence analysis and structure prediction of type II Pseudomonas sp. USM 4-55 PHA synthase and an insight into its catalytic mechanism. AB - BACKGROUND: Polyhydroxyalkanoates (PHA), are biodegradable polyesters derived from many microorganisms such as the pseudomonads. These polyesters are in great demand especially in the packaging industries, the medical line as well as the paint industries. The enzyme responsible in catalyzing the formation of PHA is PHA synthase. Due to the limited structural information, its functional properties including catalysis are lacking. Therefore, this study seeks to investigate the structural properties as well as its catalytic mechanism by predicting the three-dimensional (3D) model of the Type II Pseudomonas sp. USM 4 55 PHA synthase 1 (PhaC1P.sp USM 4-55). RESULTS: Sequence analysis demonstrated that PhaC1P.sp USM 4-55 lacked similarity with all known structures in databases. PSI-BLAST and HMM Superfamily analyses demonstrated that this enzyme belongs to the alpha/beta hydrolase fold family. Threading approach revealed that the most suitable template to use was the human gastric lipase (PDB ID: 1HLG). The superimposition of the predicted PhaC1P.sp USM 4-55 model with 1HLG covering 86.2% of the backbone atoms showed an RMSD of 1.15 A. The catalytic residues comprising of Cys296, Asp451 and His479 were found to be conserved and located adjacent to each other. In addition to this, an extension to the catalytic mechanism was also proposed whereby two tetrahedral intermediates were believed to form during the PHA biosynthesis. These transition state intermediates were further postulated to be stabilized by the formation of oxyanion holes. Based on the sequence analysis and the deduced model, Ser297 was postulated to contribute to the formation of the oxyanion hole. CONCLUSION: The 3D model of the core region of PhaC1P.sp USM 4-55 from residue 267 to residue 484 was developed using computational techniques and the locations of the catalytic residues were identified. Results from this study for the first time highlighted Ser297 potentially playing an important role in the enzyme's catalytic mechanism. PMID- 17076908 TI - Literacy and recent history of diarrhoea are predictive of Plasmodium falciparum parasitaemia in Kenyan adults. AB - BACKGROUND: Malaria is one of the most serious health problems in Kenya. In 2004, the Kenya Medical Research Institute and the US Army Medical Research Unit--Kenya surveyed adults in Samburu, Malindi, and Busia districts to determine socioeconomic risk factors for infection. METHODS: Sociodemographic, health, and antimalarial data were collected along with blood for malaria testing. A smear was considered negative only if no Plasmodium falciparum parasites were observed in 100 high-powered fields. Univariate analysis was performed with Pearson's Chi square test and univariate logistic regression. A multivariate logistic regression model was then created which included only variables found to be at least marginally significant in univariate analysis. RESULTS: A total of 1,141 subjects were recruited: 238 from Samburu, 442 from Malindi, and 461 from Busia. Smear positivities for P. falciparum were 1.7% in Samburu, 7.2% in Malindi and 22.3% in Busia. Interdistrict differences were statistically significant (p < 0.001) in univariate analysis and in a multivariate logistic regression model which included district, literacy, occupation, and recent illness as independent variables. In the model, literacy and recent diarrhoeal illness were positively and at least marginally significantly associated with parasitaemia (p = 0.023 and p = 0.067, respectively). Neither age, sex, occupation, history of malaria in the previous three months, nor use of antimalarials in the previous four weeks were significantly associated with parasitaemia. CONCLUSION: While district of residence was the variable most highly predictive for parasitaemia among Kenyan adults surveyed, both a recent history of diarrhoeal illness and literacy were at least marginally statistically significant predictors. PMID- 17076909 TI - Down-regulation of transcription elogation factor A (SII) like 4 (TCEAL4) in anaplastic thyroid cancer. AB - BACKGROUND: Anaplastic thyroid cancer (ATC) is one of the most aggressive human malignancies and appears to arise mainly from transformation of pre-existing differentiated thyroid cancer (DTC). However, the carcinogenic mechanism of anaplastic transformation remains unclear. Previously, we investigated specific genes related to ATC based on gene expression profiling using cDNA microarray analysis. One of these genes, transcription elongation factor A (SII)-like 4 (TCEAL4), encodes a member of the transcription elongation factor A (SII)-like gene family. The detailed function of TCEAL4 has not been described nor has any association between this gene and human cancers been reported previously. METHODS: To investigate the role of TCEAL4 in ATC carcinogenesis, we examined expression levels of TCEAL4 in ACLs as well as in other types of thyroid cancers and normal human tissue. RESULTS: Expression of TCEAL4 was down-regulated in all 11 ACLs as compared to either normal thyroid tissues or papillary and follicular thyroid cancerous tissues. TCEAL4 was expressed ubiquitously in all normal human tissues tested. CONCLUSION: To our knowledge, this is the first report of altered TCEAL4 expression in human cancers. We suggest that loss of TCEAL4 expression might be associated with development of ATC from DTC. Further functional studies are required. PMID- 17076910 TI - Developmental genomics reaches new heights. PMID- 17076911 TI - Lipids join the post-genomic era. PMID- 17076912 TI - Thresholds for health and thresholds for illness: panic disorder versus subthreshold panic disorder. AB - BACKGROUND: There is increasing evidence that subthreshold forms of psychopathology are both common and clinically relevant. To enable classification of these subthreshold forms of psychopathology, it may be useful to distinguish not only a threshold for illness but also for health. Our aim was to investigate this with regard to panic. METHOD: Data were derived from the Netherlands Mental Health Survey and Incidence Study (NEMESIS), which is based on a large representative sample of the adult general population (18-65 years) of The Netherlands (n=7076). The Composite International Diagnostic Interview was used as a diagnostic instrument. By defining two thresholds, three groups were formed: panic disorder, subthreshold panic disorder and no-panic. These groups were compared using multinomial regression analysis, chi2 and analysis of variance. RESULTS: The 12-month prevalence of panic disorder was 2.2% while that of subthreshold panic disorder was 1.9%. Symptom profiles and risk indicators associated with panic disorder and subthreshold panic disorder were similar, and half of the risk indicators were more strongly associated with panic disorder than with subthreshold panic disorder. Subthreshold panic disorder occupied an intermediate position between panic disorder and no-panic with regard to the number of symptoms, the percentage of subjects with co-morbidity, and functioning. CONCLUSIONS: Subthreshold panic disorder is common, and seems clinically relevant, but is milder than panic disorder. These results thus support the use of a double threshold in panic. Further research should focus on the positioning of the thresholds, the course of subthreshold panic disorder and its treatment options. PMID- 17076913 TI - False memory and schizophrenia: evidence for gist memory impairment. AB - BACKGROUND: In this study we examined patients' false memory, that is memory for a non-presented event, to search for a further source of converging evidence for the impairment of semantic memory in individuals with schizophrenia. In two experiments we compared the pattern of false memory created by the Deese-Roediger McDermott (DRM) paradigm between individuals with schizophrenia and those of a normal control group. METHOD: Experiment 1 tested participants on both recall and recognition of lists of semantically related words. Experiment 2 adopted the meaning recognition test, in addition to the standard recognition test, to assess the participants' gist memory. RESULTS: Individuals with schizophrenia performed worse than normal controls on both recall and recognition of studied words. The schizophrenia patients had higher rates of false recall and false recognition for semantically unrelated words than did the normal controls, suggesting an abnormal pattern of semantic activation in the former group. More importantly, no differences were found between the two groups with regard to false recall and false recognition of semantically related words. When the participants were tested for meaning recognition, however, the schizophrenia patients gave fewer 'old' responses to non-studied semantically related words than did the control group, indicating an impaired gist memory in schizophrenia patients. CONCLUSIONS: When asked to consciously retrieve word lists, individuals with schizophrenia showed impairment not only in item-specific memory but also in gist memory. The pattern of results is consistent with the storage deficit view of semantic memory in schizophrenia. PMID- 17076914 TI - Attentional biases for angry faces in unipolar depression. AB - BACKGROUND: Past research has demonstrated that depression is associated with dysfunctional processing of emotional information. Recent studies demonstrate that a bias in the attentional processing of negative information may be an important cognitive vulnerability factor underlying the onset and maintenance of depression. However, to date, the nature of this attentional bias is still poorly understood and further exploration of this topic to advance current knowledge of attentional biases in depression seems imperative. METHOD: This study examined attentional biases for angry facial expressions presented for 1000 ms in 20 patients with major depressive disorder (MDD) and 20 non-depressed control participants (NC) matched for age and gender using an emotional modification of the Exogenous Cueing task. RESULTS: Patients with MDD showed maintained attention for angry faces compared with neutral faces. In comparison with non-depressed participants they showed a stronger attentional engagement for angry faces. In contrast, the NC group directed attention away from angry faces, more rapidly disengaging their attention compared with neutral faces. CONCLUSIONS: This pattern of results supports the assumption that MDD is characterized by deficits in the attentional processing of negative, interpersonal information and suggests a 'protective' bias in non-depressed individuals. Implications in relation to previous research exploring cognitive and interpersonal functioning in depression are discussed. PMID- 17076915 TI - What predicts poor mother-infant interaction in schizophrenia? AB - BACKGROUND: Poor clinician-rated parenting outcome and observed interactive deficits in mothers with schizophrenia admitted to a psychiatric mother and baby unit (MBU) reflect continuing concerns over the parenting capacity of this group. However, little is known about whether interaction deficits are accounted for by severity of illness or adverse social circumstances typically experienced by these mothers. METHOD: Thirty-eight women with severe perinatal illness (schizophrenia n=13; affective disorders n=25) and their infants were observed in play interaction a week prior to MBU discharge. Clinical and sociodemographic data were also obtained. RESULTS: Mothers with schizophrenia and their infants were rated to have poorer interactive behaviour than the affective disorders group. Infant avoidance of the mother was associated with a lack of maternal sensitivity and responsiveness. The deficits in mother-infant interaction found in the schizophrenia group could not be accounted for by our measures of illness severity or factors relating to adverse social circumstances. CONCLUSIONS: The results replicate and extend previous findings showing poor interactive behaviours in mothers with schizophrenia, their infants, and in the dyad, in a range of areas following clinical recovery. The findings suggest that factors other than illness duration, dose of medication, marital status and occupational status are explanatory for the interactive deficits associated with maternal schizophrenia. Parenting interventions that aim to improve maternal sensitivity need to be developed specifically for this group. PMID- 17076916 TI - Contrasting models of genetic co-morbidity for cannabis and other illicit drugs in adult Australian twins. AB - BACKGROUND: The use of cannabis and other illicit drugs (OIDs) and their co morbid misuse are frequently reported in the literature. Correlated vulnerabilities and causal or gateway influences have been implicated in this association. We investigated the source of this co-morbidity between cannabis use (experimentation, early and repeated use, and problems) and OID experimentation and problems using genetic models proposed by Neale and Kendler (American Journal of Human Genetics 1995, 57, 935-953). METHOD: In a sample of 4152 same-sex male and female adult Australian twin individuals, we fit 13 genetically informative models of co-morbidity to data on experimentation, early use, repeated use of cannabis and co-morbid OID experimentation, and to abuse/dependence (A/D) problems with cannabis and OIDs. RESULTS: Model-fitting results suggest that common genetic, shared and unique environmental factors are responsible for the association between cannabis experimentation, early use, repeated use and A/D problems and OID experimentation or problems. The liability causation model, which is a reduced form of the correlated vulnerabilities model, also fit very well. In women, we found evidence for high-risk cannabis experimenters and repeated users to be at increased risk for OID experimentation, despite being below the risk threshold on the liability distribution for OID experimentation (extreme multiformity). CONCLUSIONS: Co-morbid cannabis and OID use and misuse are due partly to a common predisposition to substance use disorders. Putative causal effects could not be ruled out. These models warrant further research, so that features of the correlated vulnerabilities model and the gateway models can be studied jointly in a single series of adaptive nested models. PMID- 17076917 TI - Cognitive self-therapy for chronic depression and anxiety: a multi-centre randomized controlled study. AB - BACKGROUND: Non-professional treatment programmes are presumed to relieve the extensive need for care of anxiety and depression disorders. This study investigates the effectiveness of cognitive self- therapy (CST) in the treatment of depression or generalized anxiety disorder. METHOD: Patients (n=151) were randomized to receive CST or treatment as usual (TAU) in a trial lasting for 18 months, measuring symptoms (SCL-90; main outcome), social functions, quality of life and utilization of care. RESULTS: Patients in both conditions improved significantly, but no difference was found between the conditions. Reduction of symptoms, improvement of social functions and medical utilization were maintained at the end of the 18 months. Medical care utilization (therapist contact and hospitalization) was lower for CST than for TAU. No suicides occurred. CONCLUSIONS: Cognitive self-therapy is likely to decrease the need for care of chronic depression and anxiety disorders, but it has not been proven to be more effective than treatment as usual. PMID- 17076918 TI - Postpartum psychosis: two cohorts compared, 1875-1924 and 1994-2005. AB - BACKGROUND: There has been a long-standing debate as to whether postpartum psychoses are distinct from other psychoses. While the outcomes of postpartum psychosis are in general thought to be good, the disorder is linked to a high rate of suicide. METHOD: We have utilized a database of 3872 admissions to the North Wales Asylum during the period 1875-1924 to extract data on the prevalence, course and clinical features of postpartum psychoses during this period. We have collected first admissions for postpartum psychosis between 1994 and 2005 in North West Wales to establish a current incidence rate for the disorder. RESULTS: The incidence of psychoses with a first onset in the postpartum period in North West Wales has fallen in the modern period, while the incidence of postpartum psychoses in women with a pre-existing mental illness remains the same. Some features of the clinical picture and course of postpartum psychoses differ from other psychoses, but may be gender rather than disorder linked. CONCLUSIONS: These findings suggest that psychoses with their first onset in the postpartum period may be vanishing. If replicated, this would support claims that these disorders are distinct from other disorders. Alternately, if regarded as affective disorders, establishing the basis for the apparent decline in frequency of these disorders may have implications for other affective disorders. PMID- 17076919 TI - Clinically defined vascular depression in the general population. AB - BACKGROUND: Vascular depression is regarded as a subtype of depression, especially in, but not entirely restricted to, the elderly, characterized by a specific clinical presentation and an association with (cerebro)vascular risk and disease. It could have major implications for treatment if subjects at risk for such a depression could be easily identified by their clinical presentation in general practice. METHOD: We studied the symptom profile of depression in subjects with and without vascular risk factors in two large Dutch community based studies, the Rotterdam Study and the Amsterdam Study of the Elderly (AMSTEL). RESULTS: We could not confirm the specific symptom profile in depressed subjects with vascular risk factors in either of the two cohorts. Depressed subjects with vascular risk factors showed more loss of energy and more physical disability than those without vascular risk factors. However, presumed specific symptoms of vascular depression, namely psychomotor retardation and anhedonia, were not significantly associated with any of the vascular risk indicators. Loss of energy was significantly associated with myocardial infarction and peripheral arterial disease. CONCLUSIONS: In these two large community-based studies we identified some differences between vascular and non-vascular depressed subjects but found no evidence for a specific symptom profile of vascular depression as previously defined. PMID- 17076920 TI - Subtype analysis of Cryptosporidium parvum isolates from calves on farms around Belgrade, Serbia and Montenegro, using the 60 kDa glycoprotein gene sequences. AB - Cryptosporidium parvum is composed of multiple subtypes, which appear to be zoonotic or anthroponotic. Therefore, in order to understand the molecular epidemiology of this species and to control the zoonotic transmission, it is essential to identify the isolates at the subtype level. In the present study, 60.2% of 103 examined dairy calves from 10 farms in the Belgrade area were positive for Cryptosporidium infection by PCR, and all of them were identified as C. parvum by PCR-RFLP of SSUrRNA and COWP genes. Eighteen C. parvum isolates selected randomly from 9 positive farms were classified by molecular phylogenetic analysis of the 60 kDa glycoprotein gene sequences into 3 subtype allele families, IIa and IId, which are known to be zoonotic, and a new family IIj. Furthermore 10 isolates in IIa were classified into 3 subtypes IIaA16G1R1b, IIaA18G1R1, IIaA20G1R1, 2 in IId into IIdA18G1b, and 6 in IIj into IIjA16R2 and IIjA17R2. The last 2 subtypes in IIa and 2 subtypes in IIj were new subtypes. These results suggest that C. parvum isolates harboured in calves in the Belgrade area have zoonotic potential and C. parvum is genetically diverse only in a limited area. This is the first molecular epidemiological report about Cryptosporidium infection in Serbia and Montenegro. PMID- 17076921 TI - Host ecology and variation in helminth community structure in Mastomys rodents from Senegal. AB - We studied patterns of variation in parasite communities of 2 closely related species of Mastomys rodents. These 2 species live in sympatry in South-eastern Senegal, but differ drastically in their habitat choice. We asked (a) whether the host species have the same parasites; (b) whether there is any observable pattern relative to the host species/habitat type in the structure of parasite communities; (c) whether the variability in parasite community for each host species is related to habitat characteristics. We analysed 220 and 264 individuals of each host species, sampled respectively in 10 and 11 trap sites. Twenty parasite taxa were recorded, and the majority were nematodes. Between-host species comparisons showed that helminth communities were slightly more diversified in M. natalensis. Many parasite species were found in both Mastomys. However, various helminth taxa varied in frequency and abundance between host species. Within each host species, helminth diversity, prevalence and/or abundance of some parasites were correlated with habitat or host population factors that may influence parasite life-cycles, such as village structure, or the presence/absence of a pool. Our results suggest that habitat characteristics have a strong impact on helminth community structure. PMID- 17076922 TI - Alternatives to anthelmintics for the control of nematodes in livestock. AB - Efficient and welfare-friendly livestock production demands the control of nematode infection. Current control measures rely upon anthelmintic treatment but are threatened by the widespread evolution of drug-resistance in parasite populations. Several methods have been advocated to control nematodes without relying on effective anthelmintics. These include grazing management, biological control, nutritional supplementation, vaccination, and genetic approaches. Each method has its advantages and disadvantages. There are several grazing management schemes that can reduce the severity of infection but they are insufficient on their own to control infection. Biological control includes the use of predatory fungi to control nematode populations and the use of pasture species that can reduce the intensity of infection. Fungi can control nematodes but the current requirement for daily feeding means that this approach will be most useful for animals that are handled daily. Feeding supplementary protein can control nematode infection. The method is simple but can be expensive and may not be cost effective for some marginal enterprises. Genetic approaches include the use of resistant breeds and selective breeding. Some breeds will thrive in conditions that kill animals from other breeds but substitution of resistant breeds is not always feasible. Selective breeding is effective and inexpensive but requires a high level of expertise. The most appropriate method or set of methods to minimize the adverse consequences of nematode infection may vary among farms. PMID- 17076923 TI - Molecular and phylogenetic characterization of Cryptosporidium and Giardia from pigs and cattle in Denmark. AB - The genetic diversity of Cryptosporidium spp. and Giardia duodenalis from dairy cattle and pigs in Denmark was determined in the present study. Faecal samples from 1237 pigs and 1150 cattle originating from 50 sow herds and 50 dairy herds, respectively, were analysed for the presence of the two parasites by immunofluorescence microscopy. A large proportion of the (oo)cyst containing samples were selected for molecular characterization. Sequencing and phylogenetic analysis of the 18S rDNA locus and/or the HSP70 gene of 183 pig and 154 cattle isolates of Cryptosporidium revealed the presence of C. suis, pig genotype II, C. parvum (cattle genotype), C. bovis, Cryptosporidium deer-like genotype and a novel C. suis-like genotype. For both cattle and pigs, a host age-related change in distribution of species/genotypes was observed. The zoonotic C. parvum (cattle genotype) was most prevalent in young calves. For Giardia, 82 and 145 isolates from pigs and cattle, respectively, were analysed at the 18S rDNA locus and/or the gdh gene. Giardia isolates belonging to the zoonotic Assemblage A was found in both young and older calves, as well as in weaners and piglets, whereas cows seemed to be infected purely by isolates of the livestock group, Assemblage E. PMID- 17076924 TI - Molecular detection and characterization of piroplasms infecting cervids and chamois in Northern Spain. AB - Wildlife can act as reservoir of different tick-borne pathogens of veterinary and zoonotic importance. To investigate the role of wild ruminants as reservoir of piroplasm infection, 28 red deer, 69 roe deer and 38 chamois from Northern Spain were examined by reverse line blot (RLB) hybridization. The survey detected a prevalence of 85.7% in red deer, 62.3% in roe deer and 28.9% in chamois. Four different piroplasms were identified: Theileria sp. OT3 (previously described in sheep) as the most prevalent (85.7% in red deer, 46.4% in roe deer and 26.3% in chamois); Theileria sp. 3185/02 (previously described in a red deer in Central Spain) more abundant in red deer (53.6%) than in roe deer (10.1%) but absent from chamois; Babesia divergens detected in 6 roe deer; Theileria ovis present in 1 chamois. Mixed infections (Theileria sp. OT3 and Theileria sp. 3185/02) were only found in red and roe deer. Sequencing analysis of the 18S rRNA gene confirmed the RLB results and showed 99.7% identity between Theileria sp. 3185/02 and T. capreoli, suggesting that they are the same species. Tick distribution and contact of wild ruminants with domestic animals are discussed in terms of piroplasm infection. The results suggest that a considerable number of wildlife ruminants are asymptomatic carriers that may serve as reservoirs of the infection posing a serious concern in terms of piroplasmosis control. PMID- 17076925 TI - Transstadial and transovarial persistence of Babesia divergens DNA in Ixodes ricinus ticks fed on infected blood in a new skin-feeding technique. AB - Although Babesia divergens is the the principal confirmed zoonotic Babesia sp. in Europe, there are gaps in our knowledge of its biology and transmission by the tick Ixodes ricinus. In order to reproduce the part of the parasite cycle that occurs in the vector, an in vitro animal skin feeding technique on blood containing in vitro cultivated B. divergens was developed. Parasite DNA was detected in all samples of salivary glands of nymphs and adults that had fed on parasitized blood as larvae and nymphs, respectively, indicating acquisition as well as a transtadial persistence of B. divergens. PCR performed on eggs and larvae produced by females that had fed on parasitized blood demonstrated the existence of a transovarial transmission of the parasite. Gorging B. divergens infected larvae on non-infected gerbils showed persistance of the parasite over moulting into the resulting nymphs. These results indicate that the parasitic stages infective for the vector (i.e. the sexual stages) can be produced in vitro. To our knowledge, this is the first report of artificial feeding of I. ricinus via membrane as well as in vitro transmission of B. divergens to its vector. The opportunities offered by the use of such a transmission model of a pathogen by I. ricinus are discussed. PMID- 17076926 TI - Habitat and transmission--effect of tidal level and upstream host density on metacercarial load in an intertidal bivalve. AB - Transmission of parasites may be mediated by their habitat, consisting of abiotic and biotic components. I investigated the effect of 2 important habitat components in intertidal ecosystems, tidal level (abiotic) and density of upstream hosts (biotic), on the transmission of trematode cercariae to cockle (Cerastoderma edule) hosts. A field survey showed no general trend in metacercarial loads of cockles regarding tidal level but species-dependent reactions. Parasites originating from Littorina littorea (Himasthla elongata, Renicola roscovita) showed highest infection levels in the low intertidal while parasites originating from Hydrobia ulvae (H. continua, H. interrupta) showed highest infection levels in the mid-intertidal. This reflected the density of upstream hosts at both tidal levels and positive relationships between the density of upstream hosts and metacercarial load in cockles suggested the biotic habitat component to be the dominant factor in transmission. This was confirmed by a field experiment, manipulating tidal level and the density of infected upstream snail hosts. While tidal level had no significant effect on the number of metacercariae of H. elongata acquired by cockles, the effect of upstream host density was strong. In conclusion, although tidal level usually is a very important abiotic habitat component in intertidal ecosystems leading to conspicuous zonation patterns in free-living organisms, it seems of minor importance for trematode transmission. In contrast, the biotic component upstream host density is suggested to be the dominant predictor for trematode transmission to second intermediate hosts. Assessing the relative importance of abiotic and biotic habitat components in transmission is vital for the understanding of transmission processes in the field. PMID- 17076927 TI - Subcellular distribution of the Entamoeba histolytica 140 kDa FN-binding molecule during host-parasite interaction. AB - Entamoeba histolytica trophozoites recovered from the host-parasite interface during abscess development obtain different stimuli compared with long-term cultured cells. In order to have a better understanding about the mechanisms in which the 140 kDa fibronectin (FN)-binding molecule (EhFNR) is involved during the invasive process, we decided to compare the regulation process of this molecule among long-term cultured trophozoites, FN-stimulated trophozoites, and trophozoites recently recovered from a liver abscess. A cDNA clone (5A) containing a fragment of the EhFNR that shows identity to the C-terminal region of the intermediate galactose lectin subunit Igl, was selected with a mAb (3C10). Identity of EhFNR with Igl was confirmed by immunoprecipitation with 3C10 and EH3015 (against the Gal/GalNAc intermediate subunit) mAbs. The 3C10 mAb was used as a tool to explore the modulation of the amoebic receptor (EhFNR). Our results showed specific regulation of the EhFNR in FN-interacted amoebas, as well as in trophozoites recovered at different stages of abscess development. This regulation involved mobilization of the receptor molecule from internal vesicles to the plasma membrane. Therefore, we suggest that in the host-parasite interface, the EhFNR (Igl) plays an important role in the adhesion process during abscess development. PMID- 17076928 TI - Experimental study on heat transmission to the vestibule during CO2 laser use in revision stapes surgery. AB - We studied the transmission of heat to the vestibule during revision stapes surgery with a piston in situ, using a CO2 laser, in an in vitro model. A type K thermocouple was placed around the medial end of stainless steel and fluoroplastic wire pistons in a 'vestibule' filled with saline. The effect of laser hits on fluoroplastic wire and stainless steel stapes prostheses was investigated. The effect of introducing a vein graft to seal the stapedotomy was also examined. Greater temperature rises occurred with stainless steel than with fluoroplastic wire pistons. The addition of the vein graft reduced heat transmission. Application of the CO2 laser to fluoroplastic wire pistons, using the power settings suggested by the manufacturer, is not likely to damage the inner-ear structures. Application of 6 W laser energy to stainless steel pistons can potentially disturb the inner-ear function. PMID- 17076929 TI - Sialolithiasis in a residual Wharton's duct after excision of a submandibular salivary gland. AB - Treatment of salivary stones includes both surgical and non-surgical techniques. Surgical approaches range from excision of the sialolith, for those near the duct orifice, to removal of the affected salivary gland and its associated duct, for stones near the hilum of the gland. We present a case of two sialoliths triggering an acute infection in a residual Wharton's duct, 12 years after the removal of the associated submandibular gland. Excision of the sialoliths and treatment of the infected duct via sialodochoplasty was successfully performed in this patient. If the Wharton's duct is not removed with the associated submandibular gland, the potential for infection and continuous growth of dormant calcifications exists. We also address the aetiology, pathogenesis, and management of patients with sialolithiasis in the absence of a major salivary gland. PMID- 17076930 TI - Otoplasty: experience with a modification using a drill, and literature review. AB - More than 200 techniques have been described for correction of prominent ears, indicating that there is no single, widely accepted procedure that has been adopted by most surgeons. This article presents a simplified surgical method for correction of prominent ears. One hundred and twenty-eight otoplasties were performed on 70 patients using the described technique. The main modification of the technique was the use of a diamond burr drill to thin the cartilage posteriorly. Good aesthetic results were obtained in most patients. PMID- 17076931 TI - Ameloblastoma: a rare nasal polyp. AB - Ameloblastoma is an odontogenic neoplasm of enamel organ type tissue which does not undergo transformation to the point of enamel formation. We present the second case in the English literature of maxillary ameloblastoma that presented with nasal obstruction and rhinorrhoea, and the first to be excised using a combined maxillotomy and endoscopic ethmoidectomy. The patient had no previous dental history. The unusual presenting symptoms, as well as the highly destructive nature of these lesions when arising in the maxilla, make them worthy of consideration in the differential diagnosis of nasal and maxillary masses. We discuss the clinical features, pathology and management of these lesions and review the literature. PMID- 17076932 TI - Management of benign inverted sinonasal papilloma avoiding external approaches. AB - Even though endoscopic removal of inverted papillomas has gained popularity, many studies advocate supplementary external approaches. The impact of including the current surgical staging system into the pre-operative clinical and radiological assessment has not been systematically evaluated. We present our experience with total endoscopic management of inverted papillomas and compare the accuracy of the pre-operative predicted extent of surgery, with the actually performed surgery. From 1997 to 2005 data from 51 patients with inverted papillomas were prospectively collected and subsequently reviewed. All have been operated on endoscopically without an external approach. The overall recurrence rate was 3.9 per cent. Pre-operative prediction of extent of surgery was accurate in 26 of 51 (51 per cent). The main reasons for the inaccurate pre-operative prediction were the variable sizes and locations of the inverted papilloma bases, particularly in the maxillary sinus and the frontal recess. Our results encourage us to recommend endoscopic management as the standard treatment of benign inverted papillomas. PMID- 17076933 TI - Image-guided surgical microscope with mounted minitracker. AB - A new image-guided microscope using augmented reality overlays has been developed. Unlike other systems, the novelty of our design consists in mounting a precise mini and low-cost tracker directly on the microscope to track the motion of the surgical tools and the patient. Correctly scaled cut-views of the pre operative computed tomography (CT) stack can be displayed on the overlay, orthogonal to the optical view or even including the direction of a clinical tool. Moreover, the system can manage three-dimensional models for tumours or bone structures and allows interaction with them using virtual tools, showing trajectories and distances. The mean error of the overlay was 0.7 mm. Clinical accuracy has shown results of 1.1-1.8 mm. PMID- 17076934 TI - The efficacy of the dopamine D2/D3 antagonist tiapride in maintaining abstinence: a randomized, double-blind, placebo-controlled trial in 299 alcohol-dependent patients. AB - In this investigation, the hypothesis was tested whether the selective dopamine D2/D3 receptor antagonist tiapride is effective in maintaining abstinence after detoxification in alcohol-dependent patients. The rationale of the study was based on the relevance of the dopaminergic system for addictive behaviour as well as some preliminary studies. A multi-centre, randomized, double-blind, placebo controlled, parallel-group study was conducted. A total of 299 detoxified alcohol dependent patients (ICD-10: F10.2) received either tiapride (300 mg/d) or placebo over a 24-wk study period. Subjects with severe comorbid psychiatric disorder such as schizophrenia or Wernicke-Korsakoff syndrome were excluded. Primary outcome variable was the time to first relapse with relapse defined as any alcohol consumption after detoxification. Data analysis was done with Kaplan Meier estimates with log-rank test (one-sided, p<0.05). Tiapride was not superior to placebo in maintaining abstinence. The time to first relapse was 71 d in the tiapride group and 92 d in the placebo group (log-rank test, p=0.9895). Relapse rate was higher in the intervention group (54.4%) than in the control group (40.7%). Like the dopamine antagonist flupenthixol, tiapride was not effective in maintaining alcohol abstinence. Regarding the high success rate in the placebo group the influence of psychosocial treatment in studies investigating drug effects on the course of alcohol dependence has to be considered. PMID- 17076935 TI - Discriminative stimulus properties of the selective norepinephrine reuptake inhibitor, reboxetine, in rats: a characterization with alpha/beta-adrenoceptor subtype selective ligands, antidepressants, and antagonists at neuropeptide receptors. AB - Although little information is available concerning discriminative stimulus (DS) properties of antidepressants, rats can be trained to recognize the selective norepinephrine (NE) reuptake inhibitor, reboxetine (2.5 mg/kg i.p.). By analogy to reboxetine (effective dose50, 1.1), 'full' (80%) substitution dose50 was obtained with the NE reuptake inhibitors, nisoxetine (4.9), nomifensine (0.5) and BW1555,U88 (1.0). Full substitution was also attained with the NE/serotonin (5 HT) reuptake inhibitors, S33005 (0.3), venlafaxine (4.8) and duloxetine (26.8), and the tricyclics, imipramine (2.5) and clomipramine (2.9). In contrast, the 5 HT reuptake inhibitors, citalopram, sertraline and paroxetine (all >2.5), and the 5-HT reuptake inhibitors/5-HT2 receptor antagonists, nefazodone and trazodone (both >10.0), did not substitute for reboxetine. The 'atypical' antidepressants, mirtazapine (>10.0) and mianserin (>2.5), similarly failed to substitute. DS properties of reboxetine were dose-dependently blocked by the alpha1-adrenoceptor (AR) antagonists, prazosin (inhibitory dose50, 0.3) and WB4101 (0.5), but resistant to the alpha2-AR antagonists, atipamezole (>0.63), idazoxan (>2.5) and RX821,002 (>0.08), and to the beta1-AR and beta2-AR antagonists, betaxolol (>2.5) and ICI118,551 (>10.0). Interestingly, the neurokinin-1 receptor antagonist, GR205,171, stereospecifically substituted for reboxetine (1.1) compared to its less active isomer, GR226,206 (>10.0). The corticotrophin-releasing factor-1 antagonists, DMP695 (>40), CP154,526 (>10.0) and SN003 (>40.0), and the melanin concentrating hormone-1 antagonist, SNAP-7941 (>40.0), failed to substitute for reboxetine. In conclusion, DS properties of reboxetine are mimicked by antidepressants recognizing NE transporters, and require functionally intact alpha1-ARs for their expression. The neurokinin-1 antagonist, GR205,171, mimics the interoceptive properties of reboxetine, possibly reflecting its elevation of extracellular levels of NE in corticolimbic structures. PMID- 17076936 TI - Neonatal basolateral amygdala lesions affect monoamine and cannabinoid brain systems in adult rats. AB - There is evidence for neurodevelopment disturbances in schizophrenia. In rats, a neonatal basolateral amygdala lesion induces behavioural features in adults reminiscent of the symptomatology of schizophrenia. Dopamine plays a key role in the pathogenesis of schizophrenia, and cannabis use has been implicated in the risk for developing schizophrenia. The effects of an excitotoxic, bilateral basolateral amygdala lesion on postnatal days 7 or 21 were compared when the rats were adult. The behavioural response to a novelty challenge and the level of dopamine receptors and cannabinoid receptors in the brain using in-vitro autoradiography was determined. In brain tissue punches concentrations of monoamines and metabolites were determined by high-performance liquid chromatography. The neonatal lesion, but not the later lesion induced behavioural hyperactivity and biochemical effects. The neonatal lesion reduced the density of dopamine D2-like, but not D3-, and less markely D1-like receptors and increased dopamine turnover. These effects were observed in the mesolimbic, but not in the striatal regions. In contrast, density of cannabinoid receptors was increased in the striatal, but not the mesolimbic regions of these animals. Noradrenergic neurotransmission was reduced in both regions. The present findings contribute to the idea that the neonatal basolateral amygdala lesion induces features in adults reminiscent of the neurodevelopmental disturbances in schizophrenia, with a focus on the amygdala-prefrontal cortex-nucleus accumbens circuit. PMID- 17076937 TI - CO2 challenge induced HPA axis activation in panic. AB - The hypothalamic-pituitary-adrenal axis (HPA axis) plays a critical role in stress management. Involvement of this physiological axis in the underlying mechanisms of panic disorder (PD) has been suggested. Studies using 35% CO(2) inhalation to provoke panic found no evidence for robust increases in cortisol levels in PD. However, cortisol levels alone may not be conclusive, as this hormone is merely the end product of a complex physiological axis. Sixteen PD patients and 16 healthy control subjects underwent a 35% CO(2) inhalation and a placebo inhalation on separate days according to a fixed order, double-blind design. Both serum and salivary cortisol, as well as adrenocorticotropic hormone (ACTH) were measured at regular time intervals. Cortisol and ACTH levels increased in the patient and control groups following 35% CO(2) inhalation. The magnitude of the increase was similar in patients and controls despite marked differences in anxiety. This study is the first to document a clear HPA response following 35% CO(2) inhalation in both PD patients and controls. This effect occurs independently of the specific panicogenic properties of the CO(2) challenge. It remains to be clarified whether panic is initially accompanied by major HPA axis activation or whether other stress-responsive systems underlie panic. PMID- 17076938 TI - Outbreak of norovirus illness associated with a swimming pool. AB - On 3 February 2004, the Vermont Department of Health received reports of acute gastroenteritis in persons who had recently visited a swimming facility. A retrospective cohort study was conducted among persons attending the facility between 30 January and 2 February. Fifty-three of 189 (28%) persons interviewed developed vomiting or diarrhoea within 72 h after visiting the facility. Five specimens tested positive for norovirus and three specimen sequences were identical. Entering the smaller of the two pools at the facility was significantly associated with illness (RR 5.67, 95% CI 1.5-22.0, P=0.012). The investigation identified several maintenance system failures: chlorine equipment failure, poorly trained operators, inadequate maintenance checks, failure to alert management, and insufficient record keeping. This study demonstrates the vulnerability of recreational water to norovirus contamination, even in the absence of any obvious vomiting or faecal accident. Our findings also suggest that norovirus is not as resistant to chlorine as previously reported in experimental studies. Appropriate regulations and enforcement, with adequate staff training, are necessary to ensure recreational water safety. PMID- 17076939 TI - Rubella seroprofile of the Italian population: an 8-year comparison. AB - The objective of this study is to evaluate how increasing MMR infant vaccination coverage in recent years has modified the epidemiology of rubella in Italy. A cross-sectional population-based seroprevalence study of rubella antibodies was conducted on 3094 sera, in 2004, and results were compared with data obtained by the same method in 1996. The overall proportion of rubella-seropositive individuals was found to be significantly higher in 2004 with respect to 1996 (84.6% vs. 77.4%). However, an increase in seropositivity was observed only in the 1-19 years age groups. Recent increases in childhood MMR vaccination coverage, therefore, have not had an impact on seroprevalence in women of childbearing age, over 5% of whom remain susceptible to rubella. Preconception screening and postpartum vaccination of susceptible women are fundamental if the WHO target of less than one case of congenital rubella syndrome per 100,000 live births is to be attained. PMID- 17076940 TI - Transmission of bovine leukaemia virus within dairy herds by simulation modelling. AB - In Argentina, bovine leukaemia virus (BLV) infection is common in dairy herds. The country currently has a National Voluntary Control Programme but relatively few farms have enrolled. However, there is increased interest from authorities and farmers to implement regional compulsory programmes but there is scarce quantitative information of the transmission of BLV in cattle herds. This information is a prerequisite to develop effective BLV control strategies. Mathematical modelling offers ways of integrating population-level knowledge and epidemiological data to predict the outcomes of intervention scenarios. The purpose of the current paper is to gain understanding about the dynamics of the transmission of BLV in dairy herds from Argentina by simulation and to compare various BLV transmission models and select the one that is most appropriate. The hypothetical herd is conceptually described in terms of BLV status as a population of individuals that are protected by maternal antibodies (M), that are susceptible (S), that are in the latent period (E) or that are infectious (I). BLV is spread by horizontal and vertical transmission. We used an age-structured population model and within-herd transmission was simulated by Monte Carlo techniques. The next-generation approach has been used for the systematic computation of the basic reproduction ratio (R0). Parameter values for disease transmission were derived from previously published data; rates of entry, exit or transition between age groups were calculated based on our previous study, observational data, expert opinions and literature. With these parameter values the probability of a minor outbreak was estimated to be 10%, the probability of extinction was estimated as <0.001% and the expected time to extinction as more than 80 years. The probability of a minor outbreak and changes in prevalence were different when the index case was an adult cow compared to introduction by a heifer. Prediction of prevalences from MSI models fit the data satisfactorily. R0 was estimated as 9.5. The sensitivity analysis on R0 showed that all measures directed to reduce the transmission rate are potentially effective given operational control measures. An important prediction of these models is that, even in a relatively small, closed dairy herd, the time-scale for a BLV outbreak may be as long as several years and within-herd control of BLV requires intensive efforts. PMID- 17076941 TI - Modification of the Hormann technique of hyoid suspension in obstructive sleep apnoea. AB - Hyoid suspension is a procedure to stabilise the retrolingual space in obstructive sleep apnoea. Using the Hormann technique, a steel wire is slung around the body of the hyoid and fixed to the upper rim of the thyroid cartilage. It was observed, however, that the steel wire may lie very superficially to the pharyngeal mucosa. Evaluation of computed tomography (CT) scans showed a mean distance between the hyoid and the pharynx of only 3 mm. A modification is presented by threading a steel wire through a hole that is drilled through the hyoid bone. Thus pharyngeal exposure of the steel ligature and possible perforation of the mucosa is avoided. PMID- 17076942 TI - Chaperones in ENT out-patients; the patients' perspective. AB - Chaperones are used in order to avoid misunderstandings that can lead to false accusations. Consultations and examinations may be a minefield especially if the doctors are accused of sexual misconduct. The allegations may have irreparable effects on their reputation and may lead to removal from the medical register and to criminal charges being brought. We present the results of a questionnaire from 85 patients, exploring patient experience in ENT out-patients particularly with respect to examination of the ear under the microscope. This procedure necessitates close contact between the patient and doctor for several minutes and there may be inadvertent contact with intimate areas on the patient. Up to 30 per cent of patients expressed a preference for a chaperone during the ENT consultation. There was noticeably a greater proportion of men than women patients stating a preference for a chaperone during the consultation. Interestingly, there was a strong positive correlation between the presence of friends/relatives and the preference for a chaperone. Patients should be given the freedom to choose for or against the presence of a chaperone in clinic consultations. The most commonly predicted scenario requiring a chaperone is with the male doctor and female patient. This study shows that based on patient feedback, this scenario, although common is not the exclusive area in which chaperones should be used. PMID- 17076943 TI - Non-arteritic anterior ischemic optic neuropathy (NAION) and phosphodiesterase type-5 inhibitors. AB - OBJECTIVE: To determine whether a causative relationship exists between non arteritic anterior ischemic optic neuropathy (NAION) and the use of phosphodiesterase-5 (PDE-5) inhibitors for the treatment of erectile dysfunction. METHODS: A comprehensive review of the literature was performed to identify the contemporary understanding of NAION pathophysiology, epidemiology, and occurrence in men using the oral PDE-5 inhibitors sildenafil (Viagra, Pfizer), vardenafil (Levitra, Bayer AG), and tadalafil (Cialis, Lilly-ICOS LLC) for the treatment of erectile dysfunction. RESULTS: NAION is the second most common acquired optic neuropathy in men aged 50 years and older. Risk factors for NAION, cardiovascular disease, and erectile dysfunction are shared and include age, dyslipidemia, diabetes, hypertension, and cigarette smoking. To date, less than 50 cases of NAION associated with PDE-5 use have been reported to the United State's Food and Drug Administration (FDA) and five Canadian cases alerted to Health Canada. Given the large number of men safely using these agents and a limited number of events, it is not possible to determine whether NAION is directly linked to the use of PDE-5 inhibitors, underlying cardiovascular risk factors, ocular anatomical defects, a combination of these variables, or as yet unidentified factors. CONCLUSIONS: PDE-5 inhibitors have gained widespread use for the treatment of erectile dysfunction due to their safety, efficacy, and ease of use. Their role in the pathogenesis of NAION remains controversial. Reasonable and informed consent regarding the possible but low risk of NAION with the use of sildenafil, vardenafil and tadalafil is recommended. Loss or decreased vision, whether painful or painless, demands urgent patient assessment and immediate cessation of PDE-5 inhibitor use. PMID- 17076944 TI - Urological surveillance and management of patients with neurogenic bladder: Results of a survey among practicing urologists in Canada. AB - OBJECTIVE: To determine current trends in management and surveillance of the neurogenic bladder population by Canadian urologists who routinely work with and provide care for these patients. METHODS: A questionnaire was mailed to members of the Canadian Urological Association. The assessment and follow-up of upper and lower urinary tract function in neurogenic bladder patients, their optimal frequency and management of related infections were the topics of inquiry. RESULTS: One hundred and five of 602 urologists responded, for a response rate of 18%. Twenty-three (22%) of the respondents did not treat neurogenic bladder patients. Four out of five urologists who treated these patients favored a yearly renal ultrasound for routine surveillance of the upper urinary tract and routinely performed urodynamic studies for evaluation of the lower tract. Only a relatively small percentage used videourodynamics. Clean intermittent catheterization (CIC) was preferred for the management of neurogenic bladder in patients with emptying difficulties. Asymptomatic urinary tract infection (UTI) was preferably not treated. Symptomatic UTI was treated with antibiotics for 7 to 10 days by about 75% of the responding urologists, while 23% of female patients and 9% of male patients were treated for less than 7 days. Although most of the urologists had access to established treatment modalities, like CIC education and bladder augmentation, only 30% made use of botulinum toxin injections. CONCLUSIONS: This study confirms that most urologists in Canada, working with neurogenic bladder patients, follow principles reported in the literature regarding the need for evaluation, surveillance, and management of the urinary tract. However, there is no consensus on the specific methods used for surveillance of the urinary system. The results emphasize the need for clear guidelines in this field of urology in Canada. PMID- 17076945 TI - Effective analgesia and decreased length of stay for patients undergoing radical prostatectomy: Effectiveness of a clinical pathway. AB - OBJECTIVES: To assess the impact of a clinical pathway (CP) on length of stay (LOS), complications, readmission rates, and patient satisfaction for patients undergoing a radical retropubic prostatectomy (RRP). MATERIALS AND METHODS: A standardized CP for all patients undergoing RRP was developed and implemented. Post-operatively, patients enrolled in the CP received oral ibuprofen and acetaminophen analgesia, with oral and subcutaneous narcotics available for breakthrough pain. Patients enrolled in the CP were compared to a pre-CP historical cohort. Patients were asked to complete a short, validated satisfaction questionnaire 10 days post-operatively. RESULTS: Sixty-eight consecutive patients underwent a RRP following CP implementation and were compared to a historical cohort of 147 pre-CP patients. Median LOS decreased by 50% (4 days versus 2 days, p < 0.0001) while complication and readmission rates were unchanged. Patient satisfaction was high in all domains. Overall, 29.4% of patients treated within the CP required no narcotic analgesia during their admission. CONCLUSIONS: The implementation of a CP for patients undergoing a RRP is a simple and effective method for reducing LOS without compromising complication, readmission rates or patient satisfaction. PMID- 17076946 TI - Evaluation of mechanical bowel preparation methods in urinary diversion surgery. AB - OBJECTIVES: We performed the first prospective, randomized, multi-center comparison of overall quality and patient tolerability of polyethylene glycol (PEG) and sodium phosphate (NaP) solution for mechanical bowel preparation prior to urinary diversion surgery. METHODS: Between 2001 and 2003, 36 patients at six institutions underwent major urological reconstructive surgery incorporating small intestine (35 radical cystectomy with urinary diversion and 1 bladder augmentation). Patients were prospectively randomized to receive either oral polyethylene glycol (group 1, n = 16) or sodium phosphate (group 2, n = 20) for mechanical bowel preparation prior to surgery, according to our multi institutional IRB-approved protocol. All patients completed a questionnaire the morning of surgery to assess the tolerability and side effects of each agent. Quality of the bowel preparation was recorded based on intraoperative findings of the attending surgeon, who was blinded to the preparation method. RESULTS: Both bowel cleansing regimens were safe and well tolerated. Patient-reported ease of use and subjective incidence of side effects were statistically similar in the two groups, and a statistically non-significant trend to more bloating in the PEG group was also noted (p = 0.085). Surgeon-scored overall quality of preparation adequacy revealed no significant differences between oral sodium phosphate and polyethylene glycol solutions (p = 0.555). Postoperative complications were rare for each bowel preparation agent. CONCLUSIONS: Performance characteristics of oral sodium phosphate and polyethylene glycol bowel preparations appear to be similar. Each method is safe, efficacious, and well-tolerated when used prior to urinary diversion surgery. The cost for the NaP preparation was $1.40 versus $19.70 for the PEG bowel preparation. Sodium phosphate may have a slight advantage because of its convenience and economic advantage. PMID- 17076947 TI - Prostatic intraepithelial neoplasia in TURP specimens and subsequent prostate cancer. AB - PURPOSE: Prostatic intraepithelial neoplasia (PIN) is considered as a precursor lesion for adenocarcinoma of the prostate. Most data supporting this relationship comes from the short-term follow-up of patients with repeated biopsies. We report a study in which patients were followed-up for 11 years to assess the relationships between the presence of high grade PIN, low grade PIN, and atypical adenomatous hyperplasia (AAH) and the subsequent occurrence of prostate cancer. MATERIALS AND METHODS: For 601 men treated by TURP in 1990-1993, prostate specimens were reviewed to assess the presence of high grade PIN, low grade PIN, and AAH. Incidental carcinoma was observed in 67 men. Follow-up of the 534 men without incidental prostate cancer was conducted until December 2003 and 24 new prostate cancers were diagnosed. Multivariate regression models were used to assess the relationships between PIN and AAH and prostate cancer on both cross sectional and prospective data. RESULTS: High grade PIN (odds ratio (OR) = 6.16, 95% confidence interval (CI): 3.28-11.58), low grade PIN (OR = 3.06, 95% CI: 1.45 6.46), and AAH (OR = 2.06, 95% CI: 1.00-4.29) were significantly associated with incidental prostate cancer. In the prospective study, only high grade PIN was associated with a statistically significant increased risk of prostate cancer: hazard ratio = 3.12, 95% CI: 1.15-8.49. CONCLUSION: Although high grade PIN, low grade PIN, and AAH were all associated with incidental prostate cancer, the long term prospective study showed that only high grade PIN was a significant determinant of the subsequent occurrence of prostate cancer. PMID- 17076948 TI - Modified anatrophic nephrolithotomy: A useful treatment option for complete complex staghorn calculi. AB - INTRODUCTION: Management of complete staghorn calculi represents a challenging problem for urologists. We describe our technique and clinical experience with modified anatrophic nephrolithotomy in patients harboring large, extensively branched staghorn calculi. MATERIALS AND METHODS: From October 1996 to February 2005 twenty-six patients with complete staghorn calculi defined as filling the entire collecting system or at least 80% of it, were treated employing a modification of the classical anatrophic nephrolithotomy technique. The mean patient age was 46 (range 16-70) years and the mean stone size was 3150 (range 1375-4800) mm2. Intra-operative data, complications and stone-free rates were recorded. Long-term follow-up was completed in 22 patients with a mean duration of 38 (range 12-96) months. Renal function was evaluated by 99mTc dimercapto succinic acid renal scintigraphy before and 6 months after treatment. RESULTS: The mean ischemia time was 36 (range 20-45) minutes, mean operative time was 195 (range 170-235) minutes and the mean blood loss was 475 (range 300-750) ml. Length of hospital stay averaged 8.8 days. One patient developed significant hematuria requiring renal angiography and embolization of a pseudoaneurysm. Overall, 22 patients (85%) were rendered stone-free at discharge while 23 patients (88%) were observed to be stone-free after 3 months. Long-term follow-up demonstrated recurrent stone fragments less than 4 mm in three patients. Isotope studies revealed that renal function remained unchanged in 55%, improved in 32% and became worse in the small number of remaining patients. CONCLUSIONS: Modified anatrophic nephrolithotomy is a valuable treatment option for patients with complete staghorn calculi. Because of its efficacy, safety and simplicity we believe that the use of this surgical procedure is warranted in patients with a large, extensively branched, complex renal stone burden. PMID- 17076949 TI - Laparoscopic excision of para-aortic ectopic pheochromocytoma. AB - Ectopic pheochromocytomas (paragangliomas) are rare tumors that are traditionally treated with open surgery. We present our technique of such a tumor treated laparoscopically and review the safety, feasibility and advantages of this approach. PMID- 17076950 TI - Subtotal urethral duplication. AB - Urethral duplication is considered a rare congenital anomaly. The exact embryology is still controversial. A patient with subtotal urethral duplication is presented and the technique utilized to excise the anomalous structure is detailed. PMID- 17076951 TI - Fetus in fetu. AB - Fetus in fetu (FIF) is a very rare condition, with a reported incidence of one in 500,000 live births. It most likely represents a monozygotic diamniotic twin that implants itself and grows within the body of its normal karyotypically identical sibling, which typically manifests as a fetiform abdominal mass in a newborn or infant. The mass is located in the retroperitoneum in most cases, including our example, and is commonly surrounded by encapsulated fluid. However, FIF has been reported to occur in other locations, such as within the cranium, the scrotum, and the oral cavity. Usually only one fetus is present but very rarely multiple fetuses may also be present. Five fetuses in the cranium of a 1-day-old female infant with hydrocephalus (although three of these fetuses were composed of extremities only) had been reported. The fetus itself is incomplete, containing a variable number of identifiable organs. The lung, the liver, the adrenal gland, the pancreas, and the genital organs may be seen in the more complex specimens. The presence of a head with eyes, hair, and teeth has been reported, but most of these fetuses are anencephalic. We are presenting a FIF mimicking a solid and cystic renal mass in a 6-month-old boy. PMID- 17076952 TI - Fibrous pseudotumor of the epididymis and tunica vaginalis. AB - We present a case of fibrous pseudotumor of the epididymis and tunica vaginalis which is one of the rarest paratesticular tumors. Fibrous pseudotumor presents as a solid scrotal mass with normal serum markers. Pathological examination is the only reliable way to make the diagnosis with these tumors exhibiting characteristic macroscopic and microscopic features. It is important to recognize this entity since local excision is the treatment of choice and orchiectomy is unnecessary. PMID- 17076953 TI - Non-small cell lung carcinoma metastatic to the kidney. AB - INTRODUCTION: Renal metastasis by non-small cell lung cancer is uncommon and usually described at autopsy. CASE PRESENTATION: We report the rare case of a 77 year-old man with localized non-small cell lung carcinoma who underwent extirpative surgery and 2 years later presented with gross hematuria and was noted to have a large infiltrating renal mass on computed tomography. CONCLUSIONS: In patients with a prior history of resected localized non-small cell lung carcinoma, presence of a renal lesion should prompt the clinician to consider the possibility of metastasis. Further evaluation should consider this scenario. PMID- 17076954 TI - High-flow priapism: An overview of diagnostic and therapeutic concepts. AB - We describe the case of a 23 year-old man with high-flow priapism following blunt perineal trauma. He was treated successfully with super-selective embolization with a resorbable material (gel foam). We will review the pathophysiology, etiology, and diagnosis and treatment approaches of high-flow priapism based on a review of the medical literature. PMID- 17076955 TI - Intravesical botulinum type A toxin injection in patients with overactive bladder: Trigone versus trigone-sparing injection. AB - OBJECTIVE: Botulinum toxin type A (BTX-A) has been successfully used in the treatment of patients with overactive bladder (OAB) symptoms refractory to anticholinergic therapy, with most studies performing trigone-sparing detrusor injections. Increasing evidence suggest that sensory nerve dysfunction contributes to the pathophysiology of OAB and, for this reason, targeting the afferent innervation of the bladder trigone during injection may provide clinical benefit. MATERIALS AND METHODS: We conducted a pilot study to assess the benefit of trigonal-inclusion during BTX-A injection. A total of 40 patients with OAB refractory to anticholinergic treatment received trigone or trigone-sparing injection of BTX-A. Patients were evaluated using UDI-6 and IIQ-7 questionnaires prior to the BTX-A applications, at 3 weeks and 6 months after treatment. RESULTS: At 3-week follow-up, 15/24 (63%) and 10/16 (63%) patients showed improvement of symptoms in the trigone versus trigone-sparing groups, respectively. Combined 3-week UDI-6 and IIQ-7 scores improved from 37.3 prior to treatment to 27.4 (p < 0.05) and 35.5 to 27.2 (p < 0.05) in the trigone and trigone-sparing groups, respectively. Six-month follow-up demonstrated continued but diminished levels of symptom improvement when compared to pre-treatment and 3 week symptom scores. Improvement in symptom scores between the trigone and trigone-sparing groups was not significant. CONCLUSION: No significant difference in symptom score or treatment response was noted between the trigone and trigone sparing groups. Further study using pre- and post-operative urodynamic study is needed to evaluate possible benefit to trigonal injection in a select sensory urgency cohort, and to assess for urodynamic improvement following trigonal injection. PMID- 17076956 TI - The effectiveness and quality of routine child and adolescent mental health care outreach clinics. AB - OBJECTIVES: Little evidence has been available about the functioning and outcomes of child mental health services operating at the primary care interface. This study investigated the effectiveness and quality of one particular model of outreach clinic. DESIGN: The study used a quasi-experimental design comparing outcomes of children attending the outreach clinics with a waiting list comparison. The intervention group (n=88) was assessed at referral, four months and twelve months later. The comparison group (n=99) completed measures at referral and four months. METHOD: The main carers of participant children completed self-report measures of problem severity, impact, burden, parental stress and parent satisfaction. Clinicians provided clinical activity information. RESULTS: There were significant reductions in children's problem severity, distress and impairment, particularly for behavioural difficulties. In contrast, the results showed no effect upon the outcomes of children's emotional problems or parental stress. High ratings of satisfaction were obtained for many aspects of care. CONCLUSIONS: The children receiving care through this system of decentralised, outreach clinics derived clinical benefits that were maintained over time. The outreach clinics received strong consumer support suggesting the service offered a high level of acceptability to parents. PMID- 17076957 TI - Severity of Alzheimer's disease and effect on premorbid measures of intelligence. AB - OBJECTIVES: To assess the robustness of the National Adult Reading Test (NART), Wechsler Test of Adult Reading (WTAR), Cambridge Contextual Reading Test (CCRT), Spot the Word (STW), and a demographic regression equation in estimating premorbid ability in people with Alzheimer's disease (AD). DESIGN: A cross sectional multiple measures design. METHOD: Sixty-six probable AD participants were classified into stages of severity according to their Mini Mental State Examination (MMSE) score (minimal 24 to 28; mild 14 to 23). Their scores on the NART, WTAR, CCRT, STW and a demographic equation were compared with 32 healthy age matched controls. RESULTS: Significant between-group differences were found for the tests of reading ability (NART, WTAR and CCRT). The mild group made significantly more errors than the control and minimal groups. For the mild group, there was an advantage of putting the NART words into context (CCRT), but this group still made significantly more errors than the control and minimal groups. The NART was compared with the demographic estimate and there was a significant difference for the mild group with the demographic estimate providing a higher estimate. This was not the case for the control or minimal groups. There was no significant difference between the groups on the lexical decision task (STW). CONCLUSION: Reading of irregular words is compromised in AD for those with a MMSE score in the range of 14 to 23. A lexical decision task may provide a more accurate estimate of premorbid intelligence in those with mild AD. Further research is needed to provide evidence of STW's correlation with current ability. PMID- 17076958 TI - The role of clinical and social cognitive variables in parasuicide. AB - OBJECTIVES: The central aim of the present study was to investigate the extent to which social cognitive variables could mediate the effects of past self-harm behaviour and clinical variables on intentions to engage in deliberate self-harm (DSH) and suicidality in the next three months. In addition, we aimed to extend the application of the theory of planned behaviour (TPB) beyond distal health outcomes to a behaviour that is proximal and extreme. Design and method. A prospective study design was employed. Ninety parasuicide patients admitted via accident and emergency to a general hospital completed measures of hopelessness, depression, anxiety, past self-harm behaviour, standard TPB and group identity measures within days of an overdose. Three months later, participants were asked to complete a measure of suicidal thinking and behaviour. RESULTS: There was clear evidence that the social cognitive variables were significant predictors of intention to engage in DSH and suicidality three months later. Depression was the only clinical variable which remained significant when all variables were entered into the final model to predict intentions. Attitudes, self-efficacy and intention mediated the clinical variables-suicidality relationship. CONCLUSIONS: The TPB is a useful framework for understanding suicidal behaviour. The results extend the application of the TPB beyond distal health outcomes to a behaviour that is both proximal and extreme. Future research should explore the implications for screening assessment and suicide prevention. PMID- 17076959 TI - Screening for psychological distress using internet administration of the Hospital Anxiety and Depression Scale (HADS) in individuals with chronic fatigue syndrome. AB - OBJECTIVES: To investigate the factor structure and internal consistency of the Hospital Anxiety and Depression Scale (HADS) in individuals with Chronic Fatigue Syndrome (CFS) using an Internet administered version of the instrument. DESIGN: Between subjects. METHOD: Confirmatory factor analysis (CFA) and internal consistency analysis of the HADS was used to determine the psychometric characteristics of the instrument in individuals with CFS and a control group with data captured via an Internet data collection protocol. RESULTS: CFA revealed that a 3-factor solution offered the most parsimonious account of the data. Internal consistency estimations of the anxiety and depression subscales were found to be acceptable for both groups. The CFS group was found to have significantly higher HADS-assessed anxiety and depression scores compared with controls, however, there was also evidence found that Internet administration of the instrument may inflate HADS subscale scores as an artifact of testing medium. CONCLUSIONS: The HADS is suitable for use for screening individuals with CFS in terms of the factor structure of the instrument, however, clinicians should be aware that this instrument assesses 3 domains of affective disturbance rather than 2 as is interpreted within the current HADS anxiety and depression subscale scoring system. Researchers need also be aware that Internet administration of negative affective state measures such as the HADS is likely to inflate scores and need to ensure that comparisons between clinical groups are made with control group data gathered using the same collection methodology. PMID- 17076960 TI - The effectiveness of computerized cognitive behavioural therapy in routine care. AB - OBJECTIVES AND DESIGN: The efficacy of a Computerized Cognitive Behavioural Therapy (CCBT) package, Beating the Blues, has been demonstrated in a large randomized controlled trial. The current study tests the generalizability of this finding in a naturalistic non-randomized trial. METHOD: 219 patients with anxiety and/or depression were recruited to receive CCBT in routine care. The Clinical Outcomes in Routine Evaluation-Outcome Measure (CORE-OM) and Work and Social Adjustment scale (WSA) were administered pre-treatment, immediately on completing treatment and at 6 months post-treatment. Single-item self-report measures of anxiety and depression were also collected during each treatment session. RESULTS: Completer and intention-to-treat analysis demonstrated statistically and clinically significant improvements on the CORE-OM, WSA and in self-reported anxiety and depression. Intention-to-treat analysis indicated an average 0.29 point drop on the CORE-OM, equating to an uncontrolled pre-post effect size of 0.50. Research completers achieved an average 0.61-point drop equating to an uncontrolled pre-post size of 1.00 on the same measure. Where data was available (18%), these benefits were maintained at week 32 (6 months follow-up). CONCLUSION: CCBT can be an effective first line tool within a stepped care framework for the management of common mental health problems. PMID- 17076961 TI - Cue exposure in the treatment of alcohol dependence: effects on drinking outcome, craving and self-efficacy. AB - OBJECTIVES: The aim of the present study was to investigate beneficial effects of cue exposure treatment for alcohol dependence by contrasting it with a well established treatment approach. We hypothesized that cue exposure treatment is associated with a stronger decline of craving, a stronger increase in self efficacy and has beneficial effects on drinking behaviour after discharge. DESIGN AND METHODS: Sixty-three patients with a diagnosis of alcohol dependence were recruited from an in-patient alcohol-detoxification facility. Patients were sequentially assigned to either cue exposure or a standard cognitive-behavioural treatment. We assessed self-reports of craving and self-efficacy prior to treatment participation and at the end of treatment. Drinking behaviour was assessed in the 6-month period following discharge. RESULTS: Both treatments were associated with a reduction of self-reported craving and an increase in self reported measures of self-efficacy. A significant time x treatment interaction indicated a greater increase in self-reported measures of self-efficacy after cue exposure treatment. Measures of drinking behaviour showed clearly that both treatments were efficacious. Relapse rates and drinking-related variables were not significantly different for the two treatments at the 6-month follow-up. There was preliminary evidence that suggests that cue exposure therapy may be more effective for patients with severe alcohol dependence. CONCLUSIONS: With respect to drinking behaviour, cue exposure and standard cognitive-behavioural treatment seem to be equally effective for patients with a moderate severity of alcohol dependence. Further studies are necessary to specify criteria for differential treatment indication. PMID- 17076962 TI - Personalizing and externalizing biases in deluded and depressed patients: are attributional biases a stable and specific characteristic of delusions? AB - OBJECTIVES: The purpose of this study was to explore whether explicit and implicit attributional styles of delusional patients were associated to their clinical state, and whether attributions biases are specific to delusional psychopathology or also appear in other disorders (i.e. depression). DESIGN AND METHODS: A cross-sectional design was used. The sample consisted of 136 participants (40 acute deluded participants, 25 remitted deluded participants, 35 depressed patients and 36 normal controls). The Internal, Personal and Situational Attributions Questionnaire (IPSAQ) and the Pragmatic Inferential Test (PIT) were used to assess explicit and implicit attributional style, respectively. RESULTS: All participants, with the exception of the depressed patients group, showed an externalizing bias (EB) for negative events. Although both acute and remitted deluded patients showed a similar overall pattern of explicit attributions, the personalizing bias (PB) was significantly greater in the acute group. The magnitude of this bias, which was also found in the depressed patients, was significantly related to the patient's degree of severity, as assessed by the total BPRS score (r=.45, p<.001). The results on the implicit attributions were more equivocal, perhaps due the low reliability of the PIT. CONCLUSIONS: Attributional biases seem to be a stable characteristic of delusions. Yet, the PB might be a rather unspecific characteristic that varies with the degree of the severity of psychopathology. The implications of these findings for understanding the role of attributional biases in depression and delusion formation are discussed. PMID- 17076963 TI - Identifying potential predictors of traumatic reactions to psychotic episodes. AB - OBJECTIVES: The experience of a psychotic episode can sometimes lead to post traumatic stress disorder (PTSD) symptoms. The objective of the research was to identify candidate predictors of such negative reactions for future prospective study. We examined six predictors identified from the PTSD and psychosis literatures in a retrospective study: a history of previous trauma, a history of previous episodes of psychosis, perceived helplessness and uncontrollability at the time of the index psychotic episode, the content of persecutory delusions at episode and the perceived presence of crisis support after the psychotic episode. DESIGN: The design was a cross-sectional self-report and interview study of people with recently remitted symptoms of psychosis. METHOD: 36 individuals with delusions and hallucinations that had remitted in the past year were assessed for the presence of PTSD symptoms in reaction to their most recent psychotic episode. Measures of the potential predictors were also taken at this point and associations with PTSD symptoms tested. RESULTS: 61% of the individuals with remitted positive symptoms had a reaction to their psychotic episode that was potentially severe enough to receive a PTSD diagnosis. Higher levels of PTSD symptoms were associated with all six predictors tested. CONCLUSIONS: The study provides further evidence that negative reactions to psychotic episodes are relatively common. Clinicians may wish to assess for such symptoms. The study extended these findings by identifying a number of candidate psychological predictors of PTSD reactions such as perceptions of uncontrollability and absence of support. Prospective longitudinal studies are required to test the causal significance of these factors. More broadly, the findings indicate that traumatic stress in response to intra-psychic events such as delusions can be understood in similar ways to traumatic stress arising from physical traumas such as disasters. PMID- 17076964 TI - Content and affect in persecutory delusions. AB - OBJECTIVES: The study aimed to explore the content of persecutory delusions and its potential links with levels of affective disturbance. Detailed examinations of the phenomenology of delusional beliefs have been rare, but are important for furthering theoretical and clinical understanding. DESIGN: A cross-sectional investigation of 70 individuals with current persecutory delusions was conducted. METHODS: Taped semi-structured clinical interviews were transcribed for each participant. Using a coding frame devised for the current study, a detailed description of persecutory content was made. Scores on the Beck Depression and Anxiety Inventories, the Rosenberg Self-esteem Scale and the Psychotic Symptom Rating Scales were used as indicators of emotional distress. RESULTS: Data were gathered on the identity and type of persecutor, pervasiveness of threat and the power of the persecutor. Reliability was good. Beliefs involving multiple persecutors, human in nature and identifiable to the individual were common. For the majority threat was severe, ongoing and enduring and coupled with frequent feelings of vulnerability. Specific aspects of delusional content were found to be associated with emotional distress. For example, if participants felt more power in the face of persecution this was coupled with lower depression and higher self-esteem. CONCLUSIONS: Persecutory delusions are beliefs concerning severe threat, particularly of physical harm including death, which is personally significant, frequently involving multiple persecutors known to the individual. Depression is higher in those who felt less powerful than their persecutors. Associations, such as this, with emotional distress support a direct role for emotion in delusion formation and maintenance. It is consistent with cognitive models of delusions which emphasize the importance of considering emotional distress in the context of belief appraisal, although interpretation of the results is limited by the cross-sectional study design. Recognizing these links may in turn aid therapists in identifying aspects of beliefs that might be targeted to facilitate emotional change. PMID- 17076965 TI - A pilot study to investigate the effectiveness of emotion recognition remediation in schizophrenia using the micro-expression training tool. AB - OBJECTIVE: Emotion recognition impairments are a common feature of schizophrenia. This pilot study investigates the effectiveness of the 'micro-expressions training tool' (METT) to help improve this skill. METHOD: Twenty patients with schizophrenia and 20 healthy matched control participants completed the assessment, training and practice subsections of the METT. They were additionally evaluated pre- and post-training on an emotion-matching task (EMT). RESULTS: Both groups improved with METT training; patients with schizophrenia improved to a level that did not distinguish them from pre-trained controls (on both METT and EMT assessments). CONCLUSIONS: Patients with schizophrenia make significant improvements in emotion recognition following training with this tool, suggesting that brief remediation therapy may be a valuable adjunct to existing treatment programmes. PMID- 17076966 TI - Social support and psychological outcome in people with Parkinson's disease: Evidence for a specific pattern of associations. AB - OBJECTIVE: The aim of this study was to explore the relationship between social support and psychological functioning in people with Parkinson's disease. METHOD: 34 participants with idiopathic Parkinson's disease completed a comprehensive range of social support assessments and measures of depression, anxiety, stress, general psychological distress and positive affect. RESULTS: A clear pattern of relationships emerged, with the less satisfaction with social support, the higher the depression, anxiety and stress scores. Conversely, positive affect was related to more quantitative assessments of social support, such as the number of close relationships. CONCLUSION: The relationship between social support and psychological outcome in people with PD is complex. Furthermore, the complexity of this relationship should be addressed in any therapeutic attempts to relieve psychological distress and promote happiness. PMID- 17076967 TI - Management of cSSTIs: the role of daptomycin. AB - BACKGROUND: Skin and soft tissue infections (SSTIs) and complicated SSTIs (cSSTIs), particularly those caused by Gram-positive pathogens, are among the most common human bacterial infections. The emergence of resistance to antibiotics such as methicillin and vancomycin has compromised treatment options for these infections and stimulated the search for new antimicrobial therapies. Daptomycin, the first in a class of agents known as cyclic lipopeptides, is a novel antibiotic with potent activity against most Gram-positive pathogens, including methicillin-resistant Staphylococcus aureus. SCOPE: This review examines the novel properties of daptomycin and describes its therapeutic efficacy and tolerability, particularly in the treatment of cSSTIs. The data search strategy included identification of original research papers, review articles, meeting reports and editorials by searches of MEDLINE and references from relevant articles. FINDINGS: In vitro studies have demonstrated that daptomycin has superior bactericidal activity compared with vancomycin and the newer anti-Gram-positive agents, quinupristin/dalfopristin and linezolid. Robust, randomised, phase III clinical trials have shown daptomycin to be effective and well tolerated for the treatment of cSSTIs caused by Gram-positive bacteria, with equivalent clinical success rates and a similar safety profile to those of comparator agents. Data from these studies suggest a trend toward shorter duration of therapy and faster resolution of symptoms with daptomycin. CONCLUSIONS: Given the pressing need for new antibiotics to combat infections caused by Gram-positive organisms, and to overcome the problem of resistance to conventional antibiotics, daptomycin is a welcome addition to the treatment options for the management of cSSTIs. PMID- 17076968 TI - Does heart failure exacerbation increase response to warfarin? A critical review of the literature. AB - BACKGROUND: Numerous factors, such as other drugs, diet, and age, are well documented as altering response to warfarin. Less attention has been focused on the effect of disease states on the response to oral anticoagulants. Decompensated heart failure is reported to increase response to warfarin, but documentation is limited. OBJECTIVE: The purpose of this review is to critically examine the evidence of a possible effect of heart failure exacerbations on response to warfarin. RESEARCH DESIGN AND METHODS: A literature search was completed of the last 60 years using several databases, including PubMed, MEDLINE, EMBASE, and SCOPUS. Key terms in our search included 'warfarin' AND 'heart failure' and 'heart failure exacerbation' (or 'decompensated heart failure') AND 'effect on warfarin'. When relevant citations were found, the references cited by those authors were checked. RESULTS: Several reports from 1946-1989 suggested that decompensated heart failure increases response to oral anticoagulants. Unfortunately, these early reports have important limitations. More recent reports, since the widespread use of the international normalized ratio (INR), also suggest that heart failure exacerbations are associated with increased response to warfarin. Patient populations are small in these reports. CONCLUSIONS: Heart failure exacerbations may be associated with an increased response to warfarin and other vitamin K antagonists, but many reports are inadequate, and it appears that not all patients are susceptible to this effect. More frequent monitoring of INR in patients with decompensated heart failure is warranted. It is prudent to initiate warfarin at lower doses in patients with a history of heart failure and to monitor INR every 1-2 weeks during times of instability in ambulatory patients, and daily INRs in hospitalized patients. Given the large number of variables that impact on warfarin dose requirement, it is difficult to clearly establish the effect of decompensated heart failure on response to warfarin. Further studies must take all of these variables into account. PMID- 17076969 TI - A French cost-consequence analysis of the renoprotective benefits of irbesartan in patients with type 2 diabetes and hypertension. AB - OBJECTIVES: We performed a cost-consequence analysis in a French setting of the renoprotective benefit of irbesartan in hypertensive type 2 diabetes patients over a 25-year period. RESEARCH DESIGN AND METHODS: A previously published Markov model simulated progression from microalbuminuria to overt nephropathy, doubling of serum creatinine, end-stage renal disease and death. Three treatment strategies with analogous blood pressure control were compared: (A) control- conventionally medicated antihypertensive therapy (excluding angiotensin converting enzyme inhibitors, other angiotensin-2-receptor antagonists and dihydropyridine calcium channel blockers) initiated at microalbuminuria; (B) early irbesartan--(300 mg daily added to control, initiated with microalbuminuria) and (C) late irbesartan--(300 mg daily, initiated with overt nephropathy). Probabilities came from the Irbesartan in Reduction of Microalbuminuria-2 study, Irbesartan in Diabetic Nephropathy Trial and other sources. Clinical and economic outcomes were projected over 25 years. Annual discount rates were 3%. RESULTS: Compared to control, early use of irbesartan added (mean +/- standard deviation) 1.51 +/- 0.08 undiscounted life years (discounted: 0.94 +/- 0.05 years), while late irbesartan added 0.07 +/- 0.01 (0.04 +/- 0.01) years/patient. Early irbesartan added 1.03 +/- 0.06 discounted quality-adjusted life years (QALYs), while late irbesartan added 0.06 +/- 0.01 QALYs. Early and late irbesartan treatments were projected to save 22,314 +/- 1273 euro and 6619 +/- 820 euro/patient, respectively versus control. Sensitivity analysis showed that even over short time horizons both irbesartan treatments were superior to the control group. CONCLUSIONS: In France, early irbesartan treatment improved quality and length of life and reduced costs in hypertensive patients with type 2 diabetes and microalbuminuria. Late irbesartan therapy is beneficial, but earlier irbesartan leads to better outcomes. PMID- 17076970 TI - The onset of effect for escitalopram and its relevance for the clinical management of depression. AB - OBJECTIVE: To analyse the significance of 'onset of effect' on clinical outcome in the treatment of depression and the contribution of individual Montgomery Asberg Depression Rating Scale (MADRS) items to improvements in the MADRS total score. RESEARCH DESIGN AND METHOD: All five published placebo-controlled clinical studies in depression as per January 1, 2005, with escitalopram, were included in this pooled analysis. RESULTS: Of the 1636 patients who were randomised to either escitalopram (882) or placebo (754), 1333 completed 8 weeks of treatment (707 escitalopram and 626 placebo). A statistically significant difference (p < 0.05) between the MADRS total score responses of escitalopram and placebo treatments was observed at week 1. All 10 MADRS single items showed a significant treatment effect at week 8. For items representing core symptoms of depression (apparent sadness, reported sadness, inner tension, concentration difficulties, inability to feel, pessimistic thoughts and suicidal thoughts) the effect was detected early (week 1) and for other items (reduced sleep, reduced appetite and lassitude) the effect was detected later (week 6-8). Of the patients who showed an onset of effect (> or = 20% reduction in MADRS) after 2 weeks, and who remained on escitalopram until week 8, 63% were in remission at week 8 (mean MADRS score of 6.1). CONCLUSION: Onset of treatment response at 2 weeks is an important indicator of subsequent remission at 8 weeks. It would therefore be a reasonable clinical recommendation that if patients fail to show a measurable clinical improvement within 2 weeks, a dose increase should be considered at this time. PMID- 17076971 TI - Sildenafil citrate for erectile dysfunction in men with diabetes and cardiovascular risk factors: a retrospective analysis of pooled data from placebo controlled trials. AB - OBJECTIVE: Cardiovascular (CV) risk factors are associated with an increased risk of erectile dysfunction (ED). In men with diabetes mellitus (DM), pooled from clinical trials of sildenafil treatment for ED, this retrospective analysis determined efficacy and safety, overall and in subgroups with additional CV risk (i.e., hypertension, dyslipidemia, and smoking). RESEARCH DESIGN AND METHODS: From the manufacturer's database of worldwide research, 12-week data from men with DM were pooled from randomized, double-blind, placebo-controlled trials of flexible-dose sildenafil (25, 50, or 100 mg, PRN) for ED. MAIN OUTCOME MEASURES: Question 3 (achieving an erection), question 4 (maintaining an erection), and the Erectile Function domain of the International Index of Erectile Function; percentage of successful intercourse attempts according to patient event logs; and response to a global efficacy question (GEQ). Differences between groups were determined using logistic regression (percentage of responders according to GEQ) and analysis of covariance (all other outcomes). RESULTS: Inclusion criteria were met by 11 trials and by 974 men with DM and ED who were randomized to placebo (n = 482) and sildenafil (n = 492) within the selected trials. For all outcomes, overall and regardless of additional CV risk, the benefit was greater for sildenafil versus placebo (p < or = 0.0001), including 3-fold more men responding that sildenafil treatment improved their erections (62% vs. 18%) and a more than doubling of the mean +/- standard error percentage of successful sexual intercourse attempts (52.6 +/- 5.0 vs. 22.4 +/- 5.1). Adverse events were mild to moderate and included (sildenafil vs. placebo) headache (5% vs. 2%), flushing (7% vs. 2%), and dyspepsia (4% vs. 0%), which is consistent with the profile in the general population of men treated with sildenafil for ED. CONCLUSION: This retrospective analysis of pooled data showed that sildenafil was well tolerated and improved erectile function and intercourse success in men with ED and DM, regardless of additional CV risk factors. PMID- 17076972 TI - Duloxetine in the treatment of women with stress urinary incontinence: results from DESIRE (Duloxetine Efficacy and Safety for Incontinence in Racial and Ethnic populations). AB - OBJECTIVE: To evaluate the effectiveness and safety of duloxetine for the treatment of African-American and Hispanic women with stress urinary incontinence. RESEARCH DESIGN AND METHODS: The 10-week (a 2-week lead in period followed by 8 weeks of active treatment), open-label, multicenter study of duloxetine 40 mg twice daily included women with stress urinary incontinence or stress predominant mixed incontinence. Efficacy was measured by the median percent change from baseline to endpoint of weekly incontinence episode frequency. The primary objective assessed the treatment response in a pre specified group of women (n = 2960; 2321 Caucasian, 271 African-American, and 368 Hispanic) with similar baseline incontinence and comorbidity characteristics as the subjects enrolled in the placebo-controlled trials of duloxetine for the treatment of stress urinary incontinence. The efficacy in African-American and Hispanic women was compared with Caucasians using a predefined non-inferiority subpopulation analysis. Safety measures included adverse events, laboratory test results, and vital signs. RESULTS: All three subgroups reported significant (all p < 0.001) median percent decreases in weekly incontinence episode frequency: 65.7% (African-American), -73.0% (Hispanic), and -75.0% (Caucasian). Non-inferior efficacy was demonstrated for African-American and Hispanic women compared to the Caucasian women. Common adverse events included nausea (21.8%, 28.0%, 25.3%), dry mouth (7.7%, 11.4%, 11.9%), and fatigue (9.2%, 5.7%, 11.6%) for the African American, Hispanic, and Caucasian groups, respectively. CONCLUSION: Duloxetine was efficacious and well tolerated for the treatment of African-American, Hispanic, and Caucasian women with stress urinary incontinence. The trial design was successful in enrolling a diverse population of patients. The most important limitations include the lack of placebo control, the short study duration, and the exclusion of patients with less than seven incontinence episodes per week. PMID- 17076973 TI - A randomized, controlled, open-label trial of a single day of mebendazole versus a single dose of tinidazole in the treatment of giardiasis in children. AB - BACKGROUND: Giardia duodenalis is the most commonly detected parasite in the intestinal tract of humans and 5-nitroimidazole compounds, quinacrine and furazolidone have been used against giardiasis. However, cases refractory to treatment with these drugs are becoming more common worldwide. OBJECTIVE: To compare the efficacy and safety of mebendazole versus tinidazole in the treatment of giardiasis. RESEARCH DESIGN AND METHODS: 122 children (aged 5 to 15 years) of both sexes with confirmed Giardia duodenalis cysts or trophozoites in their stool samples were randomly separated into two groups of 61 individuals. Each group received either mebendazole 200 mg three times for 1 day or tinidazole 50 mg/kg in a single dose. The evaluation of the efficacy was based on parasitological response. Parents or legal guardians of each child were asked to provide three fecal samples on days 3, 5, and 7 after treatment completion. A child was considered to be cured if no Giardia trophozoites or cysts were found in any of the three post-treatment fecal specimens evaluated by direct wet mounts and/or after Ritchie concentration techniques. RESULTS: The frequency of cure was higher for tinidazole (81.97%) than for mebendazole (63.93%); the difference was statistically significant (p < 0.05). Transient abdominal pain was more common in children treated with mebendazole (p < 0.05), whereas loss of appetite, bitter taste, headache, vomiting, and nausea were more common in the tinidazole-treated group (p < 0.05). CONCLUSIONS: Three doses of mebendazole, in a single day, are inferior to a single dose tinidazole in the treatment of giardiasis. PMID- 17076974 TI - Retrospective analysis of transient elevations in alanine aminotransferase during long-term treatment with acetaminophen in osteoarthritis clinical trials. AB - BACKGROUND: In two recent osteoarthritis trials, alanine aminotransferase (ALT) elevations were observed more frequently in patients receiving acetaminophen 3.9 g daily than in patients receiving placebo, and the rates were higher than aminotransferase values observed in some previous osteoarthritis studies with acetaminophen. OBJECTIVE: To retrospectively analyze ALT data from McNeil osteoarthritis clinical studies involving acetaminophen in order to assess the frequency and magnitude of ALT elevations and rate of ALT resolution while patients remained on acetaminophen treatment. A review of the literature revealed a few reports of isolated aminotransferase elevations occurring during acetaminophen therapy, but these reports were not included in the analysis because they did not include enough information to evaluate the frequency, magnitude, and rate of ALT elevations while patients remained on acetaminophen treatment. RESEARCH DESIGN AND METHODS: Nine controlled clinical trials were identified in which at least one of the treatments was acetaminophen alone. Studies were included if patients had ALT and aspartate aminotransferase (AST) values obtained at baseline and an ALT value at an additional visit after initiating therapy. Seven studies met these criteria and were included in this analysis. In these studies, patients received acetaminophen 1950-4000 mg/day for 4 weeks up to 12 months. Laboratory testing was performed at weeks 0 and 4 in the three 4-week studies; at weeks 0, 2, 4, 8, and 12 in the two 12-week studies; at weeks 0, 1, 2, 4, 6, 8, and 13 in the 13-week study; and at months 0, 1, 3, 6, 9, and 12 in the 12-month study. The pooled data set consisted of patient demographics, dosing records, aminotransferase and bilirubin laboratory values, and adverse events. RESULTS: There were no reports of hepatotoxicity or hepatic failure in any acetaminophen-treated patient (n = 1530). In the seven studies, 1039 patients had both baseline AST and ALT activity < or = upper limit of the reference range and an on-treatment ALT measurement. While on long-term acetaminophen treatment, 181 of 1039 (17.4%) patients had an ALT value that exceeded the upper limit of the reference range. None of the 1039 patients had an on-treatment ALT level > 3 times upper limit of the reference range in conjunction with a serum bilirubin > upper limit of the reference range, and no patient had an ALT level > 10 times upper limit of the reference range. Of the 1039 patients, 44 (4.2%) had an on-treatment ALT level > 1.5 times upper limit of the reference range, and 31 of the 44 patients had a subsequent measurement of ALT. Of these 31 patients, 29 (93.5%) had documented resolution or decreasing ALT while on treatment. An ALT level > 1.5 times upper limit of the reference range was not associated with a higher frequency of symptoms potentially related to hepatic origin. LIMITATIONS: The studies included in this analysis were limited to McNeil studies, none of which were designed to specifically evaluate the patterns of ALT activity. Thus, the incidence of ALT elevations after any specific duration of dosing, and the temporal pattern of ALT elevations, cannot be accurately determined. In addition, methodological differences existed across the studies. CONCLUSION: This analysis involving > 1000 acetaminophen-treated patients shows that low-level, transient ALT elevations usually resolve or decrease with continued acetaminophen treatment, are unaccompanied by signs or symptoms of liver injury, and, as such, appear to be clinically insignificant. Maximum recommended daily doses of acetaminophen did not cause liver failure or dysfunction. PMID- 17076975 TI - Increasing importance of dry eye syndrome and the ideal artificial tear: consensus views from a roundtable discussion. AB - BACKGROUND: Dry eye syndrome is a highly prevalent, yet largely under diagnosed, condition that can substantially affect quality of life. Left untreated, dry eye is associated with chronic eye pain and increased risk of ocular surface disease. Current demographic changes and lifestyle factors indicate that the dry eye syndrome patient population will increase significantly, ensuring that general practitioners and ophthalmic clinicians alike will experience more patients presenting with dry eye symptoms. Greater public and practitioner awareness of emerging research, technologies, and therapies is crucial to ensuring appropriate interventions to meet specific patient needs and result in clinically favorable outcomes. ROUNDTABLE ASSEMBLY: In August 2005, a team of ocular surface experts convened for a 1-day roundtable session to discuss the latest information on diagnosing and treating dry eye syndrome and real-world issues in artificial tear therapy, including preservative use. ROUNDTABLE DISCUSSION: The discussion centered on the mild to moderate dry eye patient and critical features of the ideal artificial tear, which are preservative-free formulation, protection from microbial contamination, cost-effective, non-blurring, and easy to use. Products that match this profile have the advantage of being able to benefit the myriad of patients who comprise the dry eye syndrome population. Ocular surface health should always remain a top priority. Preferred Practice Pattern Dry Eye Syndrome Medical Treatment guidelines should be modified to recommend the use of preservative-free formula artificial tear products for all levels of dry eye conditions in consideration of the medical benefit they offer to dry eye syndrome sufferers. CONCLUSION: The growing prevalence of dry eye syndrome demands increased attention. Further research, enhanced diagnostic tests, increased use of preservative-free artificial tear formulations as first-line therapy, greater patient-practitioner interaction, and patient education are warranted. PMID- 17076976 TI - Asthma control in Switzerland: a general practitioner based survey. AB - BACKGROUND: Achievement of optimal asthma control is the goal of the Global Initiative for Asthma (GINA) guidelines. METHODS: In a survey involving 281 physicians, asthma control was assessed using the Juniper asthma control questionnaire (ACQ); physicians were also asked to judge patients' asthma control subjectively. RESULTS: In total, 2127 patients were included. Follow-up was available in 1893 (89%) patients (885 females). The mean time between visits was 62 +/- 29.3 days; mean age was 45 years (+/- 19 years) and 30% were smokers. Well controlled asthma was found in 298 patients (16%). Smokers were less likely to have well-controlled asthma (smokers 12% vs. non-smokers 18%). Physicians assessed asthma control to be good in 292 patients (15%), sufficient in 504 (27%), insufficient in 954 (50%) and poor in 137 (7%) patients. Of the 292 patients assessed by their physicians as 'good asthma control', only 142 (49%) were confirmed as 'well-controlled' by the ACQ. At the first visit, 1308 (69%) patients were pre-treated with any inhaled corticosteroids (ICS). Pretreatment with leukotriene receptor antagonists (LTRAs) was reported in 127 patients (7%). Add-on therapy with the LTRA montelukast was the most frequent treatment adjustment at the first visit. Out of 1893 patients who had a follow-up visit, 298 (16%) were well controlled at the first visit and 1170 (62%) at the follow-up visit. CONCLUSION: Asthma control is insufficient in the majority of patients. Improvement of asthma control can be achieved by using objective measures such as the ACQ in regular clinical practice and adapting therapy. PMID- 17076977 TI - Assessing glucose meter accuracy. AB - BACKGROUND: Self-monitoring of blood glucose plays an important role in the management of diabetes, particularly in patients treated with insulin. Blood glucose meters must have sufficient accuracy to allow patients and clinicians to monitor glycemic control and then modify treatment accordingly. Although several standards have been proposed, a consensus about how to measure glucose meter accuracy is not available. SCOPE: An informal review of recently published articles dealing with glucose meter accuracy was conducted to provide recommendations for evaluating accuracy relevant to the practicing clinician. FINDINGS: Whether in clinical practice or in accuracy studies, the accuracy of blood glucose meters is typically determined by comparing meter results to those obtained by a clinical laboratory method on samples collected from the same patient at the same time. However, multiple sample-related, analysis-related, and data display-related factors can have an impact on accuracy determination. Clinicians need to recognize these factors and take steps to minimize their impact when assessing the accuracy of glucose meters used by their patients. CONCLUSIONS: Although modern glucose meters show variable results with respect to analytical measures of accuracy, error grid analysis demonstrates that measurements deemed clinically acceptable approach 100%. PMID- 17076978 TI - Comparing bimatoprost and travoprost in black Americans. AB - OBJECTIVE: To compare the intraocular pressure-lowering efficacy and safety of topical bimatoprost 0.03% with that of travoprost 0.004% for the treatment of black patients with open-angle glaucoma (OAG) and ocular hypertension (OHT). RESEARCH DESIGN AND METHODS: Multicenter, prospective, randomized, investigator masked trial of 94 black patients previously diagnosed with OAG or OHT. All patients completed washout of ocular hypotensive medications before study participation. Patients were assigned to either once-daily bimatoprost 0.03% or once-daily travoprost 0.004% for 3 months. MAIN OUTCOME MEASURES: The primary outcome measures were mean intraocular pressure (IOP), mean change from baseline IOP, and percentage of patients who reached a target IOP reduction. Secondary measures included ophthalmologic examination and adverse events. RESULTS: Both bimatoprost and travoprost significantly lowered IOP at all study visits (p < 0.001). Bimatoprost provided mean IOP reductions from baseline that ranged from 6.8 mmHg to 7.8 mmHg (27% to 31%). Travoprost provided mean IOP reductions from baseline that ranged from 6.2 mmHg to 6.9 mmHg (25% to 28%). By month 3, 85% of participants in the bimatoprost group had a mean IOP reduction of at least 20%, compared with 68% of those in the travoprost group. Furthermore, 31.9% of those in the bimatoprost group had a mean IOP reduction of more than 40% at month 3 compared with 20.9% of those in the travoprost group. There were no significant differences in biomicroscopy, ophthalmoscopy, or visual acuity. Ocular redness was the most commonly reported adverse event in both treatment groups. No serious adverse events were reported. CONCLUSIONS: Bimatoprost and travoprost each effectively lowered IOP in this population of black patients. More patients achieved clinically relevant IOP reductions with bimatoprost. PMID- 17076979 TI - Carotid intima-media thickness as a surrogate marker for cardiovascular disease in intervention studies. AB - BACKGROUND: Cardiovascular trials using clinical endpoints to assess efficacy typically require follow-up of large numbers of participants for 3-5 years. This disadvantage has encouraged the search for well-validated surrogate markers for cardiovascular disease (CVD). These markers may provide earlier indications of efficacy in trials involving fewer participants. One approach gaining interest in recent years is the measurement of atherosclerotic progression, a major underlying cause of CVD. SCOPE: This review article aims to further substantiate the evidence supporting the use of measurement of carotid intima-media thickness (CIMT) as a surrogate marker for atherosclerosis and cardiovascular risk. FINDINGS: CIMT has consistently been related to future CVD events in population studies. CIMT is significantly related with other markers for CVD risk, such as elevated levels of risk factors and presence of atherosclerosis in the coronary arteries. Furthermore, almost all lipid-lowering trials and a large number of blood pressure lowering trials have consistently shown a reduction in progression of CIMT. In addition, the ultrasound technique for measuring CIMT is safe and highly reproducible. CONCLUSION: Thus, CIMT may be used as a surrogate endpoint in clinical trials to enable the benefits of new therapies or regimens to be more rapidly translated into clinical practice. PMID- 17076980 TI - Lipid-lowering effects of colesevelam HCl in combination with ezetimibe. AB - OBJECTIVE: The primary aim of this study was to compare the effect of colesevelam HCl in combination with ezetimibe to ezetimibe monotherapy on low-density lipoprotein cholesterol (LDL-C) levels in subjects with primary hypercholesterolemia. METHODS: Subjects with primary hypercholesterolemia (N = 86) were enrolled in a multicenter, randomized, double-blind, placebo-controlled, parallel-group study. After a 4- to 8-week washout period, subjects received colesevelam HCl 3.8 g/day plus ezetimibe 10 mg/day or colesevelam HCl placebo plus ezetimibe 10 mg/day for 6 weeks. The primary efficacy endpoint was the mean percent change in LDL-C during randomized treatment. Secondary endpoints included mean absolute change in LDL-C, mean absolute and mean percent change in levels of high-density lipoprotein cholesterol (HDL-C), non-HDL-C, total cholesterol (TC), apolipoprotein (apo) A-I and apo B, and median absolute and percent changes in triglycerides (TG) and high-sensitivity C-reactive protein from baseline to end of treatment. Of the 86 subjects randomized to treatment, 85 were included in the intent-to-treat analysis. RESULTS: After 6 weeks of treatment, colesevelam HCl plus ezetimibe produced a mean percent change in LDL-C of -32.3% versus -21.4% with ezetimibe monotherapy (p < 0.0001). Colesevelam HCl plus ezetimibe was significantly more effective than ezetimibe alone at producing mean percent reductions in TC, non-HDL-C, and apo B and increases in apo A-I (p < 0.005 for all). Neither treatment regimen resulted in significant changes in median TG levels compared with baseline (p = NS). Both treatments were safe and generally well tolerated. CONCLUSIONS: Colesevelam HCl plus ezetimibe combination therapy significantly reduced mean LDL-C, TC, non-HDL-C, and apo B levels and increased apo A-I levels (p < 0.005 for all) without significantly increasing median TG levels in hypercholesterolemic subjects compared with ezetimibe alone. Although limited in that atherosclerotic coronary heart disease outcomes were not evaluated, this study demonstrated that combining colesevelam HCl with ezetimibe is a therapeutic option in hypercholesterolemic patients, such as those in whom statins are contraindicated and/or who may have intolerances to statin therapy. PMID- 17076981 TI - Improvement in quality of life with omalizumab in patients with severe allergic asthma. AB - BACKGROUND: Patients with severe persistent asthma experience daily symptoms and frequent serious exacerbations that contribute to a significant impairment of health-related quality of life (QoL). METHODS: A pooled analysis was completed of six controlled clinical trials that evaluated the effect of add-on omalizumab on asthma-related QoL in patients with severe persistent allergic (IgE-mediated) asthma. Asthma-related QoL was assessed at baseline and treatment endpoint using the well-validated Juniper Asthma Quality of Life Questionnaire (AQLQ). Change from baseline in AQLQ total score was compared between treatments using analysis of covariance methods. The percentage of patients who achieved a clinically meaningful (> or = 0.5-point) improvement in AQLQ total score was compared using the Mantel-Haenszel Chi-square test. RESULTS: The pooled patient population comprised 2548 patients (omalizumab, n = 1342; control, n = 1206), of whom 96% had severe persistent asthma according to the GINA 2002 classification. Omalizumab produced significantly greater improvements in AQLQ total score vs the control group (mean increases of 1.01 and 0.61 points, respectively; p < 0.001). In addition, significantly more omalizumab-treated patients achieved a clinically meaningful improvement in AQLQ total score than patients in the control group (66.3% vs 52.4%; p < 0.001). CONCLUSIONS: Add-on therapy with omalizumab improves QoL to a significant and clinically meaningful level in patients with severe persistent allergic asthma. PMID- 17076982 TI - Intramuscular aripiprazole or haloperidol and transition to oral therapy in patients with agitation associated with schizophrenia: sub-analysis of a double blind study. AB - OBJECTIVE: A sub-population analysis of 325 patients with agitation (Positive and Negative Syndrome Scale Excited Component [PEC] score > or = 15 and < or = 32; score of > or = 4 on > or = 2 items) associated with schizophrenia in a randomized, double-blind study investigating the efficacy and tolerability of intramuscular (IM) aripiprazole 9.75 mg, IM haloperidol 6.5 mg, or IM placebo and the transition to oral therapy. RESEARCH DESIGN AND METHODS: Over 24 h, patients could receive up to three IM injections; the second and third administered > or = 2 and > or = 4 h, respectively, after the first, if deemed clinically necessary. Following IM treatment, oral aripiprazole or haloperidol was administered for 4 days. The primary efficacy measure was the mean change in PEC score from baseline at 2 h. RESULTS: At 2 h, mean improvements in PEC scores with IM aripiprazole ( 8.0) were significantly greater versus IM placebo (-5.7; p < or = 0.01), and similar versus IM haloperidol (-8.3). Secondary efficacy measures also significantly improved with active IM treatment versus IM placebo. Continuation with oral treatment provided continued efficacy with both active treatments. The safety profiles of IM and oral aripiprazole were similar. The incidence of extrapyramidal symptom-related adverse events was 0% with IM aripiprazole, 1.6% with IM placebo and 16.5% with IM haloperidol. CONCLUSION: Intramuscular aripiprazole is effective in patients with acute agitation associated with schizophrenia, comparable to IM haloperidol, and enables convenient transfer to oral aripiprazole therapy. PMID- 17076983 TI - Collagen hydrolysate for the treatment of osteoarthritis and other joint disorders: a review of the literature. AB - BACKGROUND: There is a need for an effective treatment for the millions of people in the United States with osteoarthritis (OA), a degenerative joint disease. The demand for treatments, both traditional and non-traditional, will continue to grow as the population ages. SCOPE: This article reviews the medical literature on the preclinical and clinical research on a unique compound, collagen hydrolysate. Articles were obtained through searches of the PubMed database (www.pubmed.gov) through May 2006 using several pairs of key words (collagen hydrolysate and osteoarthritis; collagen hydrolysate and cartilage; collagen hydrolysate and chondrocytes; collagen hydrolysate and clinical trial) without date limits. In addition, other sources of information, such as abstracts presented at scientific congresses and articles in the German medical literature not available on PubMed, were reviewed and included based on the authors' judgment of their relevance to the topic of the review. FINDINGS: According to published research, orally administered collagen hydrolysate has been shown to be absorbed intestinally and to accumulate in cartilage. Collagen hydrolysate ingestion stimulates a statistically significant increase in synthesis of extracellular matrix macromolecules by chondrocytes (p < 0.05 compared with untreated controls). These findings suggest mechanisms that might help patients affected by joint disorders such as OA. Four open-label and three double-blind studies were identified and reviewed; although many of these studies did not provide key information--such as the statistical significance of the findings- they showed collagen hydrolysate to be safe and to provide improvement in some measures of pain and function in some men and women with OA or other arthritic conditions. CONCLUSION: A growing body of evidence provides a rationale for the use of collagen hydrolysate for patients with OA. It is hoped that ongoing and future research will clarify how collagen hydrolysate provides its clinical effects and determine which populations are most appropriate for treatment with this supplement. PMID- 17076984 TI - Budgetary impact of treating acute bipolar mania in hospitalized patients with quetiapine: an economic analysis of clinical trials. AB - OBJECTIVE: To present a tool that allows estimation of the budget impact of treatments for acute mania in bipolar I disorder from a US healthcare payer perspective. METHODS: Using discrete event simulation, the course of individuals is simulated beginning with hospitalization. Discharge depends on symptom level measured by the Young Mania Rating Scale (YMRS). The treatment effect is determined using time-dependent regression equations derived from trial data, and decision rules obtained from clinical experts. Outcomes include: time to response and symptom resolution; proportion of subjects reaching each outcome; number of adverse events. Costs were obtained from hospital discharge databases, the National Medicare Physician Fee Schedule and RedBook. Different scenarios are examined, each describing the proportion of subjects on the various treatments (lithium, divalproex sodium, olanzapine, risperidone, and quetiapine--monotherapy and in combination with lithium). Analyses are intention-to-treat over 100 days, corresponding to follow-up in mania trials. Despite its flexibility and structural adaptability, the model has some important limitations related to the characteristics of the clinical trials. These include focus on inpatient management of acute mania, use of the YMRS as the model driver, polypharmacy restricted to two-drug regimens, no explicit consideration of titration and dose changes, and relatively short time horizon. RESULTS: Scenarios with a greater proportion of quetiapine users (5% vs. 40% and 100%) result in a smaller impact on the healthcare budget (6912, 6277, and 5525 dollars per patient, respectively) and improvements in patient outcomes (e.g., 43%, 47%, and 54% responding at day 21; 74%, 77%, and 80% remitting by day 84). Sensitivity analyses showed that the budget impact is influenced by drug prices, discharge criteria and side-effect management. CONCLUSION: Results suggest that increased use of quetiapine for bipolar mania in the US is economically justified and improves health outcomes. In addition, this model illustrates that discrete event simulation is a useful and versatile tool for budget impact analyses. PMID- 17076985 TI - The effect of 24 months of combination statin and extended-release niacin on carotid intima-media thickness: ARBITER 3. AB - OBJECTIVE: The ARBITER 2 trial showed that extended-release niacin (ERN) when added to statin monotherapy slowed the progression of carotid atherosclerosis over 12 months. Whether longer treatment with ERN would have a greater effect on carotid intima-media thickness (CIMT) is unknown. RESEARCH DESIGN AND METHODS: We examined the long-term effects of ERN on high density lipoprotein (HDL-C) cholesterol and CIMT during 12-24 months treatment with ERN in ARBITER 2 participants who were either continued or were crossed over (from placebo) to ERN 1000 mg daily. MAIN OUTCOME MEASURES: Among 149 subjects completing ARBITER 2, 130 (88%) enrolled in ARBITER 3. The prespecified primary endpoints were the within-group change in CIMT and HDL-C in patients receiving placebo for 12 months (n = 71), ERN for 12 months (comprised of subjects from ERN treatment during ARBITER 2 (n = 78) and those crossed over to ERN from placebo after ARBITER 2 (n = 47)), and ERN for 24 months spanning ARBITER 2 and 3 (n = 57). Five subjects discontinued the study due to flushing side effects. The study was completed by 104 subjects (47 crossed over from placebo; 57 with ERN continued from ARBITER 2). RESULTS: HDL-C increased in the ERN group from 39.5 +/- 6.7 to 48.6 +/- 13.3 mg/dl (p < 0.001) along with modest reductions in LDL-C and TG. Among 125 participants treated with ERN for 12 months, there was a net regression of CIMT of -0.027 +/- 0.011 mm (p < 0.001 vs. placebo). Among 57 participants treated with ERN for 24 months, there was additional significant regression of CIMT of 0.041 +/- 0.021 mm (p = 0.001 vs. placebo). Controlling for changes in LDL and triglycerides, only changes in HDL-C were independently associated with regression of CIMT (beta = -0.25; p = 0.001). CONCLUSION: When added to statin therapy, ERN significantly increases HDL-C and induces atherosclerosis regression measured by CIMT over 24 months. Limitations to this study include its open-label design and the inability to relate CIMT effects to clinical outcomes. PMID- 17076986 TI - A large, naturalistic, community-based study of rivastigmine in mild-to-moderate AD: the EXTEND Study. AB - BACKGROUND: Previous studies in Alzheimer's Disease (AD) suggest a benefit from switching from one cholinesterase (ChE) inhibitor to another in the event of treatment failure on the index agent. This observational, open-label study sought to evaluate the efficacy of the ChE inhibitor rivastigmine on cognition, functional autonomy and behavior in patients with mild-to-moderate AD previously treated with other ChE inhibitors (switched patients) as well as in those previously ChE-inhibitor-naive (de novo users). METHODS: Patients were eligible for a switch if they experienced a lack or loss of efficacy or had experienced intolerance to prior ChE inhibitor therapy. Rivastigmine was initiated at a dose of 1.5 mg b.i.d. and titration was done as per standard medical practice. Efficacy was assessed using the mini-mental state examination (MMSE) and an abbreviated version of the Clinician's Global Impression of Change (CGI-C) at Month 3 and Month 6. Caregiver burden was also assessed at Month 6 using a self rated scale. RESULTS: Overall, 2633 subjects were enrolled in this study. The mean MMSE improved from 20.6 at baseline to 21.5 at Month 6. More patients improved than deteriorated on every domain of the CGI-C. Caregivers felt less burdened after the 6 month evaluation period. Efficacy parameters demonstrated favorable results for both de novo and switched patients, but more so in the first group. LIMITATIONS: Open-label studies have an inherent potential for bias by both the caregiver and the physician. In this study, there was also a large percentage of missing patient records for each of the follow-up visits (Months 3 and 6). CONCLUSIONS: Patients with mild-to-moderate AD switched from previous ChE inhibitor therapy to rivastigmine can obtain measurable benefits, although the treatment effect may be less than in de novo patients. Further research into switching cholinesterase inhibitors is warranted. PMID- 17076987 TI - Factors associated with the effect-size of thiazolidinedione (TZD) therapy on HbA(1c): a meta-analysis of published randomized clinical trials. AB - OBJECTIVE: To examine factors affecting the size of the HbA(1c) response to thiazolidinedione (TZD) therapy. RESEARCH DESIGN AND METHODS: Meta-analysis of randomized TZD controlled trials which were identified using PubMed, EBSCO and Sci-lit databases and were published in English. Sociodemographic and clinical data were extracted from each trial. HbA(1c) effect size was defined as either a placebo-subtracted change in HbA(1c) or a change in HbA(1c) from baseline. Weighted multivariable regression was used to examine factors associated with changes in HbA(1c). Bootstrapped smearing estimates were computed to obtain reliable estimates of HbA(1c) effect size. RESULTS: Forty-two trials yielded 60 trial arms which represented 8322 patients treated with thiazolidinediones. Weighted placebo-subtracted change in HbA(1c) was -0.99% +/- 0.02% with an average baseline HbA(1c) of 9.1% +/- 1.0%. Weighted bootstrapped smearing estimate of the placebo-subtracted change in HbA(1c) was -1.02% +/- 0.004%. After controlling for other variables, the baseline HbA(1c) level had a significant negative association with placebo-subtracted HbA(1c) change (p = 0.004) and also with change in HbA(1c) from baseline (p = 0.002). Longer trial duration was associated with greater placebo-subtracted HbA(1c) change (p = 0.01) but not with the change in HbA(1c) from baseline. The multivariable models explained 72% of the variation in placebo-subtracted HbA(1c) change. It was not possible to estimate effects of the run-in period and obesity on TZD effect size. CONCLUSION: Baseline HbA(1c) and trial duration significantly impacted the effect size of TZD therapy on HbA(1c). Age, gender, duration of diabetes and prior use of anti diabetic therapy were not associated with the TZD effect size. PMID- 17076988 TI - The use of rhinitis medications in children receiving initial controller therapy for asthma. AB - BACKGROUND: Due to common features of asthma and allergic rhinitis, a single therapeutic approach to treating both of these conditions has been proposed. OBJECTIVE: To compare and contrast the use of rhinitis medications in a group of children initiating various controller therapies for asthma. METHODS: A retrospective, observational study using an integrated managed care database of children aged 4-17 years with an initial medical claim for asthma and an initial pharmacy claim for fluticasone propionate (FP) and salmeterol in a single inhaler (FSC), FP alone, montelukast (MON), or combination FP + MON. Outcomes included the percentage of children initiating controller asthma therapy with prescriptions for non-sedating antihistamine (NSA) and intranasal corticosteroids (INCS) and the mean number of prescriptions for NSA and INCS. RESULTS: A total of 5247 children were included. The percentage of children who filled prescriptions for NSA or INCS and the mean number of prescriptions dispensed was similar among children treated with FSC, FP, MON, and FP + MON. There were no significant differences in the relative risk of dispensing either a NSA or INCS across cohorts. Observational studies are limited by their use of administrative data and lack of access to patient records. CONCLUSIONS: Children started on common asthma controller therapy are frequent users of rhinitis medications. The quantity and frequency of these medications is not different between dispensed asthma regimens. PMID- 17076989 TI - The key role of adult stem cells: therapeutic perspectives. AB - BACKGROUND: The origin, function and physiology of totipotent embryonic cells are configured to construct organs and create cross-talk between cells for the biological and neurophysiologic development of organisms. Adult stem cells are involved in regenerating tissues for renewal and damage repair. FINDINGS: Adult stem cells have been isolated from adult tissue, umbilical cord blood and other non-embryonic sources, and can transform into many tissues and cell types in response to pathophysiological stimuli. Clinical applications of adult stem cells and progenitor cells have potential in the regeneration of blood cells, skin, bone, cartilage and heart muscle, and may have potential in degenerative diseases. Multi-pluripotent adult stem cells can change their phenotype in response to trans-differentiation or fusion and their therapeutic potential could include therapies regulated by pharmacological modulation, for example mobilising endogenous stem cells and directing them within a tissue to stimulate regeneration. Adult stem cells could also provide a vehicle for gene therapy, and genetically-engineered human adult stem cells have shown success in treatment of genetic disease. CONCLUSION: Deriving embryonic stem cells from early human embryos raises ethical, legal, religious and political questions. The potential uses of stem cells for generating human tissues are the subject of ongoing public debate. Stem cells must be used in standardised and controlled conditions in order to guarantee the best safety conditions for the patients. One critical point will be to verify the risk of tumourigenicity; this issue may be more relevant to embryonic than adult stem cells. PMID- 17076990 TI - The incremental costs of recommended therapy versus real world therapy in type 2 diabetes patients. AB - BACKGROUND: The goals of diabetes management have evolved over the past decade to become the attainment of near-normal glucose and cardiovascular risk factor levels. Improved metabolic control is achieved through optimized medication regimens, but costs specifically associated with such optimization have not been examined. OBJECTIVE: To estimate the incremental medication cost of providing optimal therapy to reach recommended goals versus actual therapy in patients with type 2 diabetes. METHODS: We randomly selected the charts of 601 type 2 diabetes patients receiving care from the outpatient clinics of Massachusetts General Hospital March 1, 1996-August 31, 1997 and abstracted clinical and medication data. We applied treatment algorithms based on 2004 clinical practice guidelines for hyperglycemia, hyperlipidemia, and hypertension to patients' current medication therapy to determine how current medication regimens could be improved to attain recommended treatment goals. Four clinicians and three pharmacists independently applied the algorithms and reached consensus on recommended therapies. Mean incremental medication costs, the cost differences between current and recommended therapies, per patient (expressed in 2004 dollars) were calculated with 95% bootstrap confidence intervals (CIs). RESULTS: Mean patient age was 65 years old, mean duration of diabetes was 7.7 years, 32% had ideal glucose control, 25% had ideal systolic blood pressure, and 24% had ideal low density lipoprotein cholesterol. Care for these diabetes patients was similar to that observed in recent national studies. If treatment algorithm recommendations were applied, the average annual medication cost/patient would increase from 1525 to 2164 dollars. Annual incremental costs/patient increased by 168 dollars (95% CI 133-206 dollars) for antihyperglycemic medications, 75 dollars (57-93 dollars) for antihypertensive medications, 392 dollars (354-434 dollars) for antihyperlipidemic medications, and 3 dollars (3-4 dollars) for aspirin prophylaxis. Yearly incremental cost of recommended laboratory testing ranged from 77-189 dollars/patient. LIMITATIONS: Although baseline data come from the clinics of a single academic institution, collected in 1997, the care of these diabetes patients was remarkably similar to care recently observed nationally. In addition, the data are dependent on the medical record and may not accurately reflect patients' actual experiences. CONCLUSION: Average yearly incremental cost of optimizing drug regimens to achieve recommended treatment goals for type 2 diabetes was approximately 600 dollars/patient. These results provide valuable input for assessing the cost-effectiveness of improving comprehensive diabetes care. PMID- 17076991 TI - Using meta-regression in performing indirect-comparisons: comparing escitalopram with venlafaxine XR. AB - BACKGROUND: In the absence of well-powered, randomised, direct-comparison trials, indirect comparisons are the only option for comparing treatment strategies. Several methodologies have been developed and each has sparked criticism. Using direct comparisons of escitalopram versus venlafaxine extended release (XR), we explore the differences between the two compounds through indirect comparisons. METHODS: The CENTRAL, Medline and Embase databases were interrogated, focusing on randomized placebo-controlled clinical trials involving adult patients treated for major depressive disorder in the acute phase. Corresponding authors were contacted to reduce missing data. Effect sizes were derived from each study's primary outcome. For indirect comparisons, a global effect size was computed through meta-regression. For direct comparisons, the studies were considered separately due to missing data. Non-inferiority assessments were employed. The conclusion of the meta-regression was then compared with the conclusions made in direct comparison trials. RESULTS: Ten placebo-controlled studies--six assessing escitalopram and four assessing venlafaxine XR--and two direct comparison studies were retrieved. Escitalopram was found to be non-inferior to venlafaxine XR in both indirect and direct comparisons with results of mean -0.02 (unilateral 95% confidence interval [CI] -0.16 to infinity) and 0.23 (95% CI -0.01 to infinity), respectively. Results obtained by both indirect and direct comparisons were similar. Investigating the influence of age, gender repartition and severity at baseline suggests that results are consistent. Results were also considered robust against publication bias. CONCLUSIONS: This empirical finding suggests that escitalopram is non-inferior to venlafaxine XR. This reinforces the evidence found in direct comparisons trials. Indirect comparisons through meta-regression may be suitable to support decision-making. To fully assess its potential, further evaluation of this methodology, using other examples, is needed. PMID- 17076992 TI - Cardiovascular risk assessment in type 2 diabetes mellitus. PMID- 17076993 TI - What to inject when oral agents fail? PMID- 17076994 TI - Pramlintide: A new tool in diabetes management. AB - The amylin analogue pramlintide acts in concert with insulin to regulate glucose metabolism. It reduces postprandial hyperglycemia by suppressing postprandial glucagon secretion, regulating gastric emptying, and reducing food intake. In clinical use, pramlintide reduces postprandial glycemic excursions and improves A(1c) without the weight gain and increased risk of hypoglycemia typically seen with intensification of diabetes therapy. PMID- 17076995 TI - Is glycemic variability important to assessing antidiabetes therapies? AB - Traditionally, statistical estimation of glycemic variability includes computing standard deviation of glucose readings or related statistics (eg, M value, mean amplitude of glucose excursions, and so forth). We advocate an alternative approach using risk measures of variability, which have substantial clinical and numerical advantages. In addition, continuous glucose monitoring (CGM) data have clinically important inherent temporal structure that should be taken into consideration. Thus, temporal variability methods are discussed for the analysis and interpretation of CGM output. PMID- 17076996 TI - Is the metabolic syndrome a real clinical entity and should it receive drug treatment? AB - Metabolic syndrome represents a cluster of risk factors commonly associated with obesity and diabetes that in turn are associated with an increased risk of cardiovascular disease. The worldwide prevalence of metabolic syndrome, obesity, and diabetes is on the rise, leading to serious public health concerns surrounding the downstream consequences in the form of premature cardiovascular disease. In this review, we examine the debate surrounding the characterization and diagnosis of metabolic syndrome and discuss potential treatment options. PMID- 17076998 TI - Are sulfonylureas passe? AB - Sulfonylureas and similarly acting rapid insulin secretagogues have long been available to manage type 2 diabetes. These agents have a well understood mechanism of action and are in large part well tolerated. However, sulfonylurea therapy is unlikely to sustain adequate long-term glycemic control and has potential side effects, including hypoglycemia and weight gain. Additional concerns exist regarding detrimental effects of certain sulfonylureas on ischemic preconditioning and cardiac outcomes. Fortunately, newer agents such as glimepiride appear less likely to adversely affect the myocardium. These agents can often be used successfully as one component of combination therapy for diabetes management. PMID- 17076997 TI - How do different GLP-1 mimetics differ in their actions? AB - Glucagon-like peptide-1 (GLP-1) mimetics have been developed to overcome the pharmacokinetic limitations of GLP-1 for the treatment of type 2 diabetes. Their mechanisms of action and clinical effects appear particularly interesting because they target the main pathophysiologic mechanisms involved in type 2 diabetes. GLP 1 receptor agonists are more powerful and are particularly advantageous by their weight loss-inducing capacity, whereas dipeptidyl peptidase IV inhibitors exhibit a better tolerance profile. However, their ultimate role is still to be defined in the therapeutic strategy of type 2 diabetes. PMID- 17076999 TI - Are thiazolidinediones good or bad for the heart? AB - Type 2 diabetes is a global epidemic contributing to significant cardiovascular morbidity and mortality. The high prevalence of cardiovascular disease can largely be attributed to the metabolic syndrome with its multiple cardiovascular risk factors, including central obesity, hypertension, glucose intolerance, chronic inflammation, and dyslipidemia. The peroxisome proliferator-activated receptor-gamma agonists, the thiazolidinediones, may potentially correct the inflammatory disarray, endothelial dysfunction, dyslipidemia, and plaque vulnerability associated with diabetic cardiovascular disease through their effects on insulin resistance and fat metabolism, yet they can also exacerbate congestive heart failure. This review summarizes basic science, animal, and human data on the effects of thiazolidinediones on cardiovascular disease. PMID- 17077000 TI - Does a low-fat dietary intervention reduce CVD risk? PMID- 17077001 TI - Orlistat: Current issues for patients with type 2 diabetes. AB - Orlistat, a pancreatic lipase inhibitor, was approved by the US Food and Drug Administration (FDA) in the spring of 1999 as an adjunct to lifestyle intervention for weight loss. This paper seeks to examine current issues regarding orlistat use in patients with type 2 diabetes. There are a number of trials that demonstrate the benefits of orlistat over placebo for reducing body weight and improving other health parameters. Of some interest are the preliminary explorations of interaction on cytokine levels, where a possible cardiovascular benefit is plausible. Implications of the FDA approval of over-the counter use and the pharmaceutical development of another lipase inhibitor are also examined. PMID- 17077002 TI - Community-based programming to treat childhood obesity. AB - Overweight and obesity among children and adolescents have more than doubled in the past 30 years. This has in turn led to increases in debilitating chronic diseases in children, including type 2 diabetes mellitus. Clinicians may find community-based weight management programs to be useful adjunctive resources to assist youth and families to practice and maintain healthy nutrition and physical behaviors. PMID- 17077003 TI - What can the National Weight Control Registry teach us? AB - Given the high obesity rate and its economic burden, it is critical to better understand weight loss and maintenance. The National Weight Control Registry (NWCR) provides useful information about the strategies used by successful weight maintainers long term; recent data suggest that a diet with less food group variety will provide a lower calorie intake and that medical triggers have been associated with better initial weight loss and maintenance. The NWCR data suggest that long-term weight loss maintenance can be achieved and that health care practitioners may need to adjust weight control programs accordingly. PMID- 17077004 TI - [Construction and expression of the eukaryotic plasmids containing different subunits of human IL-12 gene]. AB - AIM: To clone and construct the eukaryotic expression plasmids containing different subunits of human IL-12 and to investigate its expression following transient transfection into COS-7 cells. METHODS: Human leukaemia cells (HL-60 and THP-1) were stimulated by DMSO, IFN-gamma and LPS, and the target genes including IL-12 p40 and p35 subunits were amplified by RT-PCR. P40 and p35 fragments were then connected and the recombinant plasmids pcDNA-p40, pcDNA-p35 and pcDNA-p40-p35 were constructed by double restriction enzymes digestion and connection. Positive plasmids were transiently transfected into COS-7 cells with siPORT XP-1 reagent. The expression of target gene was detected by RT-PCR and ELISA. RESULTS: The recombinant plasmids were confirmed by PCR amplification, double restriction enzymes digestion and DNA sequencing. Target gene expression was detected by RT-PCR and ELISA in the transfected COS-7 cells. CONCLUSION: The successful construction of the recombinant plasmids pcDNA-p40, pcDNA-p35 and pcDNA-p40-p35 provides a useful tool for further research on the immune regulatory role of IL-12. PMID- 17077005 TI - [RelB gene expression in murine bone marrow-derived mature and immature dendritic cells]. AB - AIM: To explore the expression of avian reticuloendotheliosis viral(v-rel) oncogene-related B (RelB) mRNA in vitro in murine mature and immature myeloid dendritic cells (DCs). METHODS: The bone marrow was collected from the femur and tibias of C57BL/6 mice in sterile condition. Bone marrow precursors were cultured with recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM CSF) and interleukin-4 (rmIL-4) to produce immature DCs. Immature DCs were stimulated by LPS 18 h before the end of culture to become mature. DCs' phenotype was detected by flow cytometry (FCM). RelB expression and protein level were determined by RT-PCR and immunofluorescence staining. RESULTS: The expression level of co-stimulatory molecules (CD86 and CD40) and MHC-II class molecule were low in immature DCs, whereas high in mature DCs. RelB expression was significantly higher in mature DCs than in immature DCs (P<0.01). CONCLUSION: RelB expression is closely associated with the maturative situation of DCs. Inhibition of RelB expression in DCs may induce tolerogenic DCs. PMID- 17077006 TI - [The effect of hyperoxia on transdifferentiation of type II alveolar epithelial cells in premature delivery rat]. AB - AIM: To investigate the effect of hyperoxia on the transdifferentiation of type II alveolar epithelial cells (AECII) in premature delivery rats. METHODS: AECII from fetal rat lung were cultured primarily to establish a model of cells damaged by hyperoxia. The morphological changes of the cells were observed by inverted phase contrast microscope and transmission electron microscope. AECII-specific surfactant protein C (SP-C) and AECI-specific aquaporin 5 (AQP5) were detected by immunocytochemical staining. The expression levels of SP-C, AQP5 mRNAs and their protein were detected by RT-PCR and flow cytometry. RESULTS: With the time of exposure to hyperoxia, primarily cultured AECII spread and flattened, losing lamellar bodies and microvilli. They rapidly lost their characteristics and gained some AECI appearance. With their morphological changes, AECII stopped expressing AECII-specific protein SP-C but began expressing AECI-associated protein AQP5. Compared with exposure to air at the same time in air group for 24, 48 and 72 hours, the expression rate of SP-C mRNA in positive cells and fluorescence index (FI) decreased markedly in 3 groups, but compared with exposure to air for 24 and 48 hours the expression of AQP5 increased significantly in 2 groups. The expression of AQP5 began decreasing and showed no marked difference compared with air group at the same time of for 72 hours. CONCLUSION: The transdifferentiation of AECII induced by hyperoxia may play a key role in the repair of alveolar epithelial cell injury in premature delivery rats. PMID- 17077007 TI - [The influence on the BK channel beta1 subunit expression of SO caused by high cholesterol in rabbits]. AB - AIM: To investigate the effects on protein expression of big-conductance Ca(2+) sensitive K(+) channel (BKca) beta1 subunit caused by high cholesterol in Rabbit Oddi's sphincter (SO) cells. METHODS: The rat-anti-rabbit polyclonal antiserum against beta1 subunits of BKca channel of SO cell was prepared. And the protein expression of BKca channel beta1 subunit of SO tissue was detected by semi quantitative immunohistochemical staining. RESULTS: The protein expression of BKca channels beta1 subunit of SO tissue in HC group was reduced, and there's statistically significant difference between the HC group and the control group. CONCLUSION: High cholesterol can reduce the protein expression of BK Channel's beta1 subunit in Rabbits' SO which suggests high cholesterol can affect the function of BKca channel. PMID- 17077008 TI - [Transfection of GFP mRNA in dendritic cells and analysis of some factors involved]. AB - AIM: To investigate the transfection of green fluorescent protein (GFP) mRNA in dendritic cells (DC) and analyze some factors which influence the transfection efficiency. METHODS: GFP (as a report gene) mRNA with cap was synthesized, in vitro, with mMESSAGE RNA Transcription Kit containing T7 RNA polymerase, and then the poly(A) was added to the GFP caped-mRNA by yeast poly(A) polymerase. DC were generated from the monocytes isolated from human peripheral blood by stimulation of GM-CSF and IL-4. The GFP mRNA was transfected into DC mediated by transfection reagent. The transfection efficiency and the expression levels were measured by flow cytometry. RESULTS: GFP expression in DC has been obtained by transfecting its mRNA synthesized in vitro. The transfection reagents, mRNA concentrations and cell densities have the significant effects on the transfection. The high level of transfection efficiency (up to 27%) was obtained using Transmessenger Transfection Kit with 1 microg gfp mRNA in 200 microL X-VIVO-15 serum-free medium at the cell density of 2.5x10(9)/L. CONCLUSION: The high-level transfection efficiency of gfp gene in DC could been achieved by using GFP mRNA in the optimum transfection conditions. PMID- 17077009 TI - [Effect of calpain inhibitor I on ikappaBalpha expression and cytokine secretion in RAW264.7 cells attacked with LPS]. AB - AIM: To explore the effect of calpain inhibitor I(CI-I) on ikappaBalpha expression and cytokine secretion in RAW264.7 cells after LPS attack. METHODS: RAW264.7 cells were pretreated with CI-I for 1 h and attacked with LPS. The ikappaBalpha expression and TNF-alpha and IL-6 secretion were detected by Western blot and ELISA, respectively. RESULTS: CI-I inhibited the decrease of ikappaBalpha expression in RAW264.7 cells treated with LPS. The secretion of both TNF-alpha and IL-6 secretion was notably increased after RAW264.7 cells were treated with LPS for 4 h or 8 h. CI-I and dexamethasone(DEX) inhibited these effects, and the combination of DEX and CI-I had synergistic effect. CONCLUSION: CI-I and DEX can inhibit the decrease of ikappaBalpha expression and prevent TNF alpha and IL-6 secretion in RAW264.7 cells attacked with LPS, which contributes to the alleviation of cellular injury. PMID- 17077010 TI - [Preliminary study on lymphotactin expression of monocyte-derived dendritic cells]. AB - AIM: To induce dendritic cells (DCs) from human peripheral blood monocytes in vitro and to investigate the kinetics lymphotactin (Lptn) mRNA expression. METHODS: Monocytes were isolated from normal human peripheral blood cells by density gradient centrifugation and the plastic-adherent cells were cultured with the cytokines (rhGM-CSF, rhIL-4, rhTNF-alpha). The immature and mature DCs' surface antigens CD1a and CD83 were analyzed by flow cytometry (FCM). The morphology of mature DCs induced by rhTNF-alpha was observed under electron microscope. Lptn cDNA was amplified by RT-PCR, cloned to vector pGM-T Easy and completely sequenced. The RT-PCR products of cells were separated on gel and analyzed by semi-quantification. RESULTS: The cells cultured for 7 d exhibited special dendritic morphology of mature DCs and highly expressed CD83. The sequence of cDNA was confirmed and was identical to that of NO. U23772 published in GenBank. The Lptn mRNA expression of DCs was stronger in DCs cultured for 7 d than in those for 5 d, whereas it was not detected in those for 3 d. CONCLUSION: The monocyte-derived dendritic cells can express Lptn mRNA in a maturation dependent manner. PMID- 17077011 TI - [High level expression of recombinant human sCD40L-IZ and identification of its biological activation]. AB - AIM: To clone and express the recombinant human soluble CD40 ligand(CD40L) with biological activation. METHODS: The isoleucine zipper (IZ) gene was fused at N terminal of CD40L gene coding E107-L261 contributing to form trimer. The gene coding hexahistidine was fused at the N-terminal of IZ because the sCD40L-IZ fusion protein could be purified by affinity chromatography. Then the fusion gene was amplified and cloned into expression plasmid pET30a. RESULTS: The protein sCD40L-IZ with His-tag at the N-terminal was effectively expressed in E. coli BL21 (DE3) as inclusion bodies and a denaturation and refolding procedure was performed to acquire soluble sCD40L-IZ. The fusion protein sCD40L-IZ was conveniently purified using HiTrap affinity column with above 95% purity. The gel filtration chromatography and non-reduced SDS-PAGE identified the trimeric structure of the recombinant protein. The microscope analysis showed the sCD40L IZ interacted with the membrane CD40 on XG2, a multiple myeloma cell line. CONCLUSION: The recombinant human trimeric CD40L-IZ was expressed in prokaryotic cells successfully, which has provided a basis for further study of the relationship between CD40L and apoptosis, CD40L and pathogenesis of illness. It is also useful to clinical treatment. PMID- 17077012 TI - [Analysis of in vitro delivering CD8+ T epitopes by attenuated bacteria]. AB - AIM: To analyze the efficiency of delivery for CD8(+) T cell epitopes by recombinant bacteria vectors. METHODS: The recombinant E. coli strain 13A or 25A and recombinant Salmonella typhimurium strain SL7207 expressing CD8(+) T cell epitopes of Lymphocytic Choriomeningitis Virus (LCMV) and Ovalbumin (OVA), which were fused with green fluorescent protein (GFP) marker at the C-terminal, were infected into LK(b) cells, LL(d) cells or bone marrow dendritic cells (BMDC). The efficiency of presentation for CD8(+) T cell epitopes by recombinant bacteria was analyzed by in vitro antigen presentation assay. RESULTS: After the infection of LK(b) cells, LL(d) cells or BMDC by recombinant bacteria, about 90% of cells were GFP positive. The results indicated that attenuated strain 13A, 25A and SL7207 had better invasive capacity to LK(b) and LL(d) cells, while BMDC had stronger capacity for uptaking recombinant bacteria. At 2 h post infection, CD8(+) T cell epitopes presented on the surface of those LK(b), LL(d) and BMDC cells infected by 13A (ptG2F) and SL7207(ptG2F) could be recognized by B3Z or nV1H7 T hybridoma cells, which were specific for OVA peptide p257-264 or LCMV peptide p118-132, respectively. But the efficiency of presentation for OVA or LCMV CD8(+) T cell epitope was decreased or increased respectively at 48 h post infection. However, LK(b), LL(d) and BMDC cells infected by 25A (ptG2F) did not effectively stimulate the specific B3Z or nV1H7 T cells. Furthermore, the presentation efficiency of BMDC was higher than those of LK(b) and LL(d) cells under the same condition. CONCLUSION: CD8(+) T cell epitopes can be delivered and presented to T lymphocytes by APC by attenuated bacteria in vitro. PMID- 17077013 TI - [Serum levels and clinical significance of soluble CD40 in liver disease]. AB - AIM: To measure the level of serum soluble CD40 (sCD40) in patients with acute hepatitis, hepatitis gravis and primary carcinoma of the liver, and to evaluate the relationship of sCD40 with biochemical marks and disease prognosis. METHODS: Patients with acute hepatitis (n=49) hepatitis gravis (n=22) and primary carcinoma of the liver (n=13) were studied, and serum sCD40 was determined in these patients and compared with that of healthy controls (n=44) by enzyme linked immunosorbent assay (ELISA). The binding capacity of serum sCD40 to its ligand CD40L was detected by flow cytometry (FCM) in vitro. RESULTS: Concentration of sCD40 was significantly higher in patients with liver disease than that in healthy controls (P<0.001), but no significant difference was found between the three types of liver disease (P=0.475). In the hepatitis gravis group, sCD40 concentration in dead patients was higher compared with that in the survivals (P<0.05). Level of sCD40 in patients with acute hepatitis was correlated with serum alanine transaminase (ALT) and aspartic transaminase (AST). The serum sCD40 could bind CD40L in vitro. CONCLUSION: These data suggest that sCD40 is an important serological marker in liver disease to evaluate acute injury of hepatocytes, and it shows a relevance with the prognosis of hepatitis gravis. The highly elevated level of sCD40 suggest the involvement of CD40 and its ligand CD40L in liver disease. PMID- 17077014 TI - [Study of the humoral immune suppression of recombinant human collagen type II peptide 250-270]. AB - AIM: To investigate the modulation of rhC II 250-270 peptide on special humoral immune response in the course of oral administration. METHODS: ELISA was used to determine antigen-specific antibodies in mice sera. The frequency of anti-C II and anti-C II (250-270) antibody-forming spleen cells was measured by ELISPOT. RESULTS: The level of C II- and C II (250-270)- specific IgG in serum from the mice fed with rhC II (250-270) were (0.82+/-0.02) and (0.84+/-0.04) respectively, and lower significantly than those of collagen-induced arthritis (CIA) control group. The anti-C II (250-270) antibody responses were suppressed obviously (P<0.01). The frequency of antibody-forming cells in the spleen from rhC II (250 270)-fed mice were (158+/-9 counts/well) and (181+/-10 counts/well) respectively, and also were reduced significantly when compared with that in CIA control group (247+/-16 counts/well)(P<0.05). CONCLUSION: Oral rhC II (250-270) could induce specific suppression of humoral responds in CIA. These findings together with a better understanding of the mechanisms of oral tolerance and regulation of humoral immune response in CIA, will help the development of innovative therapeutic intervention for rheumatoid arthritis. PMID- 17077015 TI - [Effect of HepG2 cells modified with CD80-IgG1 Fc fusion protein on anti-tumor immune response by lymphocytes]. AB - AIM: To investigate the anti-tumor immune response of lymphocytes elicited by HepG2 cells modified with CD80-IgG1 Fc fragment fusion protein (CD80-Fc). METHODS: HepG2 cells were modified with CD80-Fc, then the expression of CD80 on the cell surface was analyzed by flow cytometry (FCM). After mixed lymphocyte tumour cell reaction (MLTR) of the modified HepG2 cells and peripheral lymphocytes of healthy volunteers, the proliferation and cytotoxicity of the lymphocytes were tested by MTT colorimetry and LDH release assay,respectively. RESULTS: CD80-Fc could be efficiently bound on HepG2 cells. HepG2 cells modified with CD80-Fc fusion protein dramatically elicited proliferation and cytotoxicity of normal lymphocytes. CONCLUSION: CD80 may play an important role in anti-tumour immune response. HepG2 cells modified with CD80-Fc fusion protein can elicited potent anti-tumor immune response. This fusion protein provides a convenient means for further potential use in immunotherapy of tumor. PMID- 17077016 TI - [In situ analysis of distribution of immunocompetent cells in tumor's local draining lymph nodes]. AB - AIM: To observe the distribution of immunocompetent cells (ICCs) in tumor's local draining lymph nodes (LDLN) at different stages of disease including non metastasis, micro-metastasis and late metastasis. METHODS: 71 LDLN from 22 breast cancer, 28 LDLN from 7 gastric carcinoma were analysed by using catalyzed signal amplification (CSA) immunohistochemical staining. Monoclonal antibodies (mAbs) to perforin, granzyme B, CD8, CD56, CD68, S-100, CD134 and CD25 were used to detect the amount and functional change of ICCs. RESULTS: Paracortical hyperplasia and sinus histocytosis was mainly observed in tumor's LDLN. As it was reported, the density of S100(+) dendritic cells (DCs) was decreased as the draining lymph nodes became tumor-containing from the status of tumor free (P<0.05), and the morphology of DCs turned to be inactivated. As the lymph nodes were invaded by tumors, the densities of CD134(+) lymphocytes and CD25(+) cells were significantly increased (P<0.01). Furthermore, there was a significant trend of decreasing in the number of activated cytotoxic T lymphocytes (granzyme B(+) cells) in LDLN as tumor progressed from non/micro-metastasis, early metastasis to advanced stage metastasis (P<0.01). There was no obvious difference in distribution of ICC between non/micro-metastasis sentinel lymph nodes and non sentinel lymph nodes. CONCLUSION: The dynamic change of the ICCs in LDLN implied that immune response of tumor bearing host tends to be inhibited with the progress of tumors. PMID- 17077017 TI - [Effect of IL-1 receptor antagonist on cell apoptosis in rat corneal grafts]. AB - AIM: To observe the apoptosis of cells in rat corneal grafts at acute rejection phase and explore the effects of IL-1 receptor antagonist (IL-1ra) on cell apoptosis. METHODS: The penetrating corneal transplantation model was established. Corneal grafting was divided into four groups, namely, normal Wistar rat control group (no grafting, group A), isograft (Wistar rat-->Wistar rat, group B), allograft (Wistar rat-->SD rat) with normal saline treatment (group C) and allograft (Wistar rat-->SD rat) with IL-1ra treatment (group D). Cell apoptosis in corneal grafts was detected by TUNEL staining at 7 d, 10 d and 14 d after transplantation, and an automatic image analyzer was used to analyze the results, which were expressed as positive unit (PU). The changes of cellular ultrastructure in corneal grafts were observed under transmission electron microscope. RESULTS: (1)The average survival time of corneal grafts in C and D groups was (10.38+/-1.85) d and (13.56+/-1.94) d, respectively, with significant difference (P<0.01). (2)As compared with normal and non-rejected corneas, cell apoptosis and necrosis commonly existed in corneal grafts which rejection had occur. (3)In normal corneas, there were merely a very small number apoptotic cells in epithelial laminal, and apoptotic cells were found hardly in stromal laminal and endothelial cell layers. However, sporadic apoptotic cells were found in all layers of corneal grafts in B, C and D groups at 10 d after transplantation, the average PU being of no notably difference (P>0.05). Apoptosis obviously increased in nearby regions of wound and central area of corneal grafts in C and D groups, especially in C group. The apoptotic cells were distributed mainly in basal layer of epithelial cells and stroma of superficial layer. CONCLUSION: Cell apoptosis plays an important role in corneal graft rejection reaction. IL-1ra treatment can prolong the survival time of corneal grafts by means of suppression of cell apoptosis in corneal grafts. PMID- 17077018 TI - [The characterization of Th1/Th2 profile in end-stage renal disease patients and the correlation with the apoptosis of T lymphocyte]. AB - AIM: To investigate the characterization of Th1/Th2 profile in patients with end stage renal disease (ESRD) and the correlation with the apoptosis of the peripheral blood T cells; and to study the influence of different dialysis membranes on the apoptosis of T lymphocyte. METHODS: T cells from 10 non-dialyzed (ND) patients, 45 maintenance hemodialysis patients with cellulose acetate (CA) membrane, low-flux (PS-LF) and high-flux polusulfone (PS-HF) membrane, and 8 healthy volunteers (HC) were separated and stimulated with PHA for 24 hours in vitro. Then the apoptosis of T cells and supernatants levels of IFN-gamma and IL 4 were detected by Flow cytometry (FCM) and ELISA. RESULTS: In ESRD patients, the expression of Annexin V in T lymphocyte was higher than that of group HC (P<0.05), group CA was higher than group PS-HF and PS-LF (P<0.05). The level of IFN-gamma of ESRD patients was decreased significant compared with that in group HC (P<0.05), and there was negative correlation. between the Annexin V and IFN gamma. IL-4 was increased in ESRD patients (P<0.05) and it was positive correlated with Annexin V. CONCLUSION: ESRD patients showed suppressed secretion of IFN-gamma, increased secretion of IL-4 and apoptosis of T lymphocytes. PMID- 17077019 TI - [The abnormality of FcgammaRIIB1-mediated signaling and the hyperactivity of B cells from patients with systemic lupus erythematosus]. AB - AIM: To evaluate the effect of FcgammaRIIB(1) (CD32) representative self inhibitory adjustive mechanism of B cells on pathogenesis of systemic lupus erythematosus (SLE) by observing the expression characteristic and functional state of molecules on the surface of B cells from SLE patients. METHODS: The peripheral blood mononuclear cells (PBMC) were prepared by density gradient centrifugation, and the B cells were isolated from PBMC by magnetic activated cell sorting (MACS). The fluxes of intracytoplasmic calcium ([Ca(2+)](i)) of B cells activated by different activators were measured by fluorescence spectrophotometric method. The IgG production by B cells cultured with activators was assayed by ELISA. The expression levels of CD32, CD19, and IgM on the surface of B cells were measured by flow cytometry. RESULTS: (1)After B cells were stimulated with goat anti-human mu chain F(ab')(2) fragments and whole IgG respectively, the ratio of [Ca(2+)](i) response by F(ab')(2) fragments to whole IgG was significantly lower in SLE B cells compared to rheumatoid arthritis (RA)(P<0.05) or normal (P<0.01) B cells. (2)The ratio of total IgG production by B cells cultured with staphylococcal protein A (SPA) to SPA plus IgG anti-mu chain was significantly lower in SLE patients compared to RA patients or normal individuals (P<0.05). (3)There was no obvious difference in the expression of CD19, CD32, and IgM on the surface of B cells from SLE, RA patients and normal individuals (P>0.05). CONCLUSION: The inhibitory signaling abnormality of CD32 possibly contributes to the mechanism of hyperactivity of human SLE B cells. PMID- 17077020 TI - [Evaluation of the relationship between IL-18 levels in urine and parameters of renal pathological changes in patients with lupus nephritis]. AB - AIM: To evaluate the relationship between IL-18 levels in urine and parameters of renal pathological changes in patients with lupus nephritis (LN). METHODS: IL-18 levels in morning free urine and 24-hour's urine in 19 normal persons and 55 patients with LN were measured by ELISA. The correlation between IL-18 levels and parameters of renal pathological changes, namely activity index (AI) and chronicity index (CI), were analyzed by liner correlation analysis method. RESULTS: IL-18 levels in morning free urine and 24-hour's urine in LN group were elevated significantly compared with control group. In both groups IL-18 levels in morning free urine were (247.1+/-317.5) ng/L and (20.3+/-14.5) ng/L, respectively, P<0.001; those in 24-hour's urine were (192.1+/-170.1) ng/d and (21.0+/-3.8) ng/d, respectively, (P<0.001). There was close positive correlation between IL-18 levels in morning free urine and 24-hour's urine and LN patient's AI (for morning free urine: r=0.602, P<0.001; for 24-hour's urine: r=0.461, P<0.005) but there was no correlation between IL-18 levels in morning urine and 24-hour's urine and CI (P>0.05). Patients with LN were divided into three groups (high, moderate and low) according to AI value. There was distinct difference of IL-18 levels in urine among the three groups: IL-18 levels in morning urine were (69.2+/-82.7) ng/L, (193.5+/-106.1) ng/L and (580.7+/-453.1) ng/L, respectively, (P<0.001); those in 24-hour's urine were (103.5+/-141.4) ng/d, (188.8+/-124.0) ng/d and (333.1+/-183.2) ng/d, respectively. CONCLUSION: It is very simple and convenient to detect IL-18 levels in morning free urine, so it is a good method for evaluating renal pathological activity of LN. PMID- 17077021 TI - [Construction and preliminary application of a human natural phage antibody library with diversity]. AB - AIM: To construct a human natural phage single-chain antibody (scFv) library with diversity. METHODS: V(H) and V(L) genes were amplified by RT-PCR and hemi-PCR from peripheral blood lymphocytes of healthy persons. The V genes were assembled to form scFv by overlap PCR and cloned into phagemid pCANTAB-5E, and then transformed into E. coli TG1 by electroporation to construct a human natural phage scFv library. The diversity and gene family of antibody gene were analysed by sequencing and the specific antibodies against various antigens were screened through bio-panning. RESULTS: A human natural phage scFv library with diversity and 2x10(8) sink size was constructed successfully. The specific human scFvs against 5 antigens were obtained by bio-panning. CONCLUSION: A human natural phage scFv library with diversity is constructed successfully and can be applied to human antibody preparation. PMID- 17077022 TI - [Cytotoxic mechanism of anti-human death receptor 5 monoclonal antibody mDRA-6]. AB - AIM: To investigate the cytotoxic action and its mechanism of a novel anti-human DR5 monoclonal antibody (mAb mDRA-6). METHODS: The cytotoxic action of mAb mDRA-6 on Jurkat cells and the effects of inhibitors of caspase 8 and caspase 9 on apoptosis of Jurkat cells induced with mAb mDRA-6 were detected by flow cytometry. The effects of mAb mDRA-6 on the morpha of Jurkat cells was observed by fluorescence microscope. The apoptosis of Jurkat cells was detected by flow cytometry with Annexin V-FITC/PI staining. The DNA fragmentation in Jurkat cells was analysed by agrose gel electrophoresis. RESULTS: mAb mDRA-6 exerted cytotoxicity on Jurkat cells in dose-dependent and time-dependent manner. Jurkat cells treated with mDRA-6 exhibited typical apoptostic features in morphology, namely, membrane crenation, bubbling, chromatin condensation, and formation of apoptotic bodies. The flow cytometry analysis showed that phosphatidylserine (PS) was highly expressed in Jurkat cells treated with mDRA-6. Agrose gel electrophoresis indicated that DNA fragmentation occurred in Jurkat cells. Inhibitor of caspase 8 inhibited the apoptosis of Jurkat cells induced with mDRA 6 while Inhibitor of caspase 9 showed less effect. CONCLUSION: mDRA-6 may exert cytotoxicity by inducing Jurkat cell apoptosis through signal transduction pathway of death receptors, which may be a useful tool in treating tumors with DR5 as target molecule and exploring the functional domain of DR5. PMID- 17077023 TI - [Preparation of antibody against Memo protein and analysis of expression profile of Memo in mouse tissues]. AB - AIM: To prepare and characterize antibody against Memo protein and to detect the tissue distribution of Memo in mice. METHODS: Fusion protein GST-Memo was expressed and purified, and polyclonal antibody against Memo was prepared by immunizing mice. A FLAG-tagged eukaryotic expression vector pcDNA3-FLAG-Memo was constructed. The specificity of the antibody was detected by Western blot. RESULTS: An eukaryotic expression vector pcDNA3-FLAG-Memo was obtained. The polyclonal antibody was found to be specific to Memo. Memo protein was widely expressed in mouse tissues using the obtained antibody in Western blot. CONCLUSION: Antibody specific to Memo has been successfully obtained, which provides useful tool for investigation into Memo-associated mechanisms of tumor metastasis and invasiveness. PMID- 17077024 TI - [Expression of anti-keratin human Fab in Pichia pastoris and optimization of expression condition]. AB - AIM: To express human anti-keratin Fab in Pichia pastoris secretively and optimize the expression condition. METHODS: Genes of kappa chain and Fd fragment of anti-keratin antibody from the plasmids p3MH/Fab were subcloned into vectors pPIC9K and pPICZalphaA respectively. After confirmed by DNA sequence analysis, the recombinant plasmids pPIC9K/L and pPICZalphaA/Fd were transducted into the genome of GS115 Pichia pastoris using two-step integrating technology. Mut(+) multiple insert transformants were screened by G418 and Zeocin. The pH value and methanol concentration was adjusted to optimize the expression condition. RESULTS: Under the optimized expression condition, the Fab of anti-keratin antibody was efficiently secreted into the medium. Western blot assay proved that the expressed protein had specific keratin binding activity. CONCLUSION: The successful expression of the anti-keratin Fab in Pichia pastoris has laid a solid foundation for its further application. PMID- 17077025 TI - [Expression of recombinant human hemangiopoietin and preparation of its polyclonal antibody]. AB - AIM: To express the recombinant fusion protein of hemangiopoietin (HAPO) and prepare the rabbit-anti-human HAPO polyclonal antibody. METHODS: The sequence encoding HAPO was amplified by PCR and cloned into plasmid pET32c to construct recombinant prokaryotic expression system. The recombinant expression vectors were identified by enzyme digestion analysis and transformed into E. coli. The HAPO protein was purified by affinity chromatography. Rabbits were immunized with the HAPO protein, and the immune sera of rabbits were collected. Antibodies (IgG) obtained from the immune sera were purified. RESULTS: The purified HAPO protein was successfully obtained. The purified polyclonal antibody of rabbit-anti-human HAPO was also obtained from the immune sera of rabbits, and could response to human HAPO. CONCLUSION: A prokaryotic expression system of human HAPO has been prepared and the polyclonal antibody against HAPO has been prepared, which can be used to determine HAPO protein. PMID- 17077026 TI - Polyphasic classification of Alternaria isolated from hazelnut and walnut fruit in Europe. AB - Brown apical necrosis of English walnut and grey necrosis of hazelnut are destructive fruit diseases caused by a complex of opportunistic fungi including several small-spored catenulate Alternaria taxa. Thirty Alternaria isolates recovered from walnut and hazelnut fruit that were pathogenic on their respective host were compared along with type or representative isolates of A. alternata, A. tenuissima, A. arborescens, and A. infectoria using morphological and molecular criteria. Morphological examination using standardized procedures separated the walnut and hazelnut isolates into three morphological groups: the A. alternata group, the A. tenuissima group, and the A. arborescens group based upon common characteristics of the conidium and the sporulation apparatus. To evaluate genetic relationships among these groups, AFLP markers, inter simple sequence repeat (ISSR) markers, and histone gene sequence data were compared. Based upon AFLP data, the A. alternata and A. tenuissima groups comprised a single lineage, and the A. arborescens group comprised a separate lineage. ISSR data supported the grouping by AFLP data except for three isolates of the A. alternata group that clustered with the A. arborescens group. Base substitution of the H4 gene supported the discrimination of the A. arborescens group from the A. alternata and A. tenuissima groups. Tests of hypotheses based upon groupings derived from the various data sets supported the discrimination of the A. arborescens group but did not support the discrimination of the A. alternata group from the A. tenuissima group. PMID- 17077027 TI - Episodic remembering creates access to involuntary conscious memory: demonstrating involuntary recall on a voluntary recall task. AB - The idea that involuntary memory retrieval might occur during voluntary memory retrieval can be traced back to Ebbinghaus (1885/1964). However, methodological challenges have stood in the way of testing this idea. Recent diary studies of naturally occurring involuntary memories have provided insights into how this idea could be tested (Berntsen, 1998; Mace, 2005b, 2006). These studies have shown that the contents of an involuntary memory sometimes trigger additional involuntary memories, a process called chaining. It was hypothesised that chaining should also occur on autobiographical memory tasks, and this was tested in the present study by asking participants in an autobiographical memory task to report the presence of involuntary memories while they were performing the task. The results showed that involuntary memories did occur when participants were recalling the past. The results also showed involuntary memories dissociating from voluntary memories on two independent measures, thereby enhancing the credibility of the results. PMID- 17077028 TI - Remembering the past and imagining the future: a role for nonvisual imagery in the everyday cognition of blind and sighted people. AB - Images can be generated in all sensory modalities. Nevertheless, research on the everyday use of mental imagery, for example in autobiographical memory tasks, has suggested that it is only visual images that facilitate memory retrieval (e.g., Williams, Healy, & Ellis, 1999). If this is the case, individuals born without sight may be forced to rely more on verbal encoding (Goddard & Pring, 2001). This paper explores the presence and everyday role of nonvisual sensory imagery in 16 individuals with and 16 without sight. Using a cue word paradigm, contrary to previous research, Experiment 1 suggested that for both blind and sighted people, nonvisual imageries have a significant role to play in the generation of autobiographical memories. These results were reinforced by similar findings in Experiment 2, which used the same cue word method to explore the role of visual and nonvisual (auditory) imagery when generating future events. The results refute the claim that "useful" imagery in everyday tasks is exclusively visual. PMID- 17077029 TI - Event plausibility does not determine children's false memories. AB - In this paper we ask how the plausibility of an event affects the likelihood that children will develop a false memory for it. Over three interviews 6-year-olds and 10-year-olds were shown two true photos and two false photos-a plausible and less plausible event-and reported what they could remember about those events. Children also rated their confidence that the events happened, and how much they could remember about the events. By the final interview, within each age group, there were no differences in children's confidence ratings for the two false events. In addition, within each age group, the rate of false memories was the same for each event; across age groups, younger children developed more false memories than older children. PMID- 17077030 TI - Global gist extraction in children and adults. AB - The DRM paradigm was used to examine the role of global gist extraction in producing false memories in children and adults. First-graders, third-graders, and adults watched a videotape of a woman reading seven DRM lists, and then took a recognition memory test. Blocked (vs random) presentation and instructions to attend to the theme of lists were manipulated to enhance gist processing. In the first experiment, blocked presentation increased false recognition relative to random presentation in adults but not in first-graders or third-graders. In the second experiment, instructions to attend to list themes increased false recognition in third-graders and not in adults or first-graders. The results suggest a developmental pattern in which children become more adept at global gist extraction as they get older. These results are consistent with fuzzy trace theory's prediction of better gist processing as children grow older. PMID- 17077031 TI - Effects of response-stimulus interval manipulation and articulatory suppression on task switching. AB - We conducted three experiments using a list paradigm to examine how articulatory suppression and response-stimulus interval (RSI) manipulation affected task switching. Experiments 1 and 2 tested task-switching performance under a short and long RSI and three concurrent task conditions (control, articulatory suppression, and tapping) without external task cues. The results indicated that alternation had a greater effect under articulatory suppression than under the control and tapping conditions, and that articulatory suppression costs were unrelated to the RSI. In Experiment 3, an external task cue was provided with each stimulus, and the negative effect of articulatory suppression on alternation cost was eliminated. These results indicated that articulatory suppression effects did not differ between conditions of short and long RSI and that the availability of verbal representations of task information was independent of RSI length. This paper discussed the possible roles played by the phonological loop in task-switching paradigms. PMID- 17077032 TI - Children's memory for complex autobiographical events: does spacing of repeated instances matter? AB - Often, when children testify in court they do so as victims of a repeated offence and must report details of an instance of the offence. One factor that may influence children's ability to succeed in this task concerns the temporal distance between presentations of the repeated event. Indeed, there is a substantial amount of literature on the "spacing effect" that suggests this may be the case. In the current research, we examined the effect of temporal spacing on memory reports for complex autobiographical events. Children participated in one or four play sessions presented at different intervals. Later, children were suggestively questioned, and then participated in a memory test. Superior recall of distributed events (a spacing effect) was found when the delay to test was 1 day (Experiment 1) but there was little evidence for a spacing effect when the delay was 1 week (Experiment 2). Implications for understanding children's recall of repeated autobiographical events are discussed. PMID- 17077033 TI - Strategic grouping in the spatial span memory task. AB - Spatial short-term memory performance was examined in relation to participants' strategies. A total of 20 adult participants viewed and reproduced sequences of locations that varied in length (five, six, seven, or eight locations) and spatial separability (a manipulation of the configurations). In trial-by-trial self-reports, participants described five types of strategies. Chunking the spatial sequences into groups of three or four locations was the sole strategy associated with increased accuracy. Participants demonstrated considerable variability in the strategies that they selected, suggesting that cognitive resources are allocated to strategy selection, execution, and monitoring in the spatial span task. Spatially separable sequences were more accurately recalled than nonseparable sequences, independent of strategic grouping, suggesting two levels of grouping in the spatial span task. PMID- 17077034 TI - The role of free choice in memory for past decisions. AB - After choosing between different options, people tend to remember the features of the options in ways that favour the chosen alternative. The present experiment examined how limitations on freedom to choose between options affected this memory bias. Participants were given a series of two-option choices and were either allowed free choice between options or were assigned to an option. Participants assigned to an option were led to believe that either the selection was random or was made in their best interest based on their personality profile. Results indicated that the choice and best interest conditions demonstrated memory attributions that favoured their received options, whereas the assignment condition did not. These findings support the view that memory biases towards received options are not unique to free choice situations, but may stem from expectations and implicit theories about how and why the choice was made. PMID- 17077035 TI - Colon, colonography, and conservatism. PMID- 17077036 TI - Computerized diagnosis of emphysema. PMID- 17077037 TI - Diagnostic performance of computed tomography colonography in symptomatic patients and in patients with increased risk for colorectal disease. AB - PURPOSE: To evaluate the diagnostic performance (colorectal lesions) of computed tomography (CT) colonography in 111 patients, a majority of whom were at high risk for colorectal neoplasia. MATERIAL AND METHODS: After bowel preparation, CT colonography was performed, immediately followed by conventional colonoscopy. The diagnostic performance of CT colonography was analyzed relative to lesion size, histological diagnosis, and diagnostic certainty. RESULTS: The sensitivity of CT colonography increased with lesion size (P<0.001), and was 91% (21/23) for lesions > or = 10 mm. All 10 carcinomas and 86% (19/22) of adenomas > or = 5 mm were detected. Unconfirmed or false-positive CT findings were generally small and/or reported with low diagnostic certainty. The specificity of CT colonography would be 45% (30/66; 95% CI 34% to 57%) if patients with findings of any size and any diagnostic certainty were selected for follow-up, and 92% (85/92; 95% CI 85% to 96%) if only patients with CT findings > or = 10 mm classified as certain were selected. CONCLUSION: CT colonography had a high sensitivity for lesions > or = 5 mm. The diagnostic performance increased with lesion size and degree of diagnostic certainty, and was higher for adenomas. PMID- 17077038 TI - Cine magnetic resonance imaging of the small bowel: comparison of different oral contrast media. AB - PURPOSE: To evaluate several substances regarding small bowel distension and contrast on balanced steady-state free precession (bSSFP) cine magnetic resonance (MR) images. MATERIAL AND METHODS: Luminal contrast was evaluated in 24 volunteers after oral application of two different contrast agent groups leading to either bright lumen (pineapple, blueberry juice) or dark lumen (tap water, orange juice) on T1-weighted images. Bowel distension was evaluated in 30 patients ingesting either methylcellulose or mannitol solution for limiting intestinal absorption. Fifteen patients with duodeno-jejunal intubation served as the control. Quantitative evaluation included measurement of luminal signal intensities and diameters of four bowel segments, qualitative evaluation assessed luminal contrast and distension on a five-point scale. RESULTS: Quantitative and qualitative evaluation of the four contrast agents revealed no significant differences regarding luminal contrast on bSSFP images. Quantitative evaluation revealed significantly lower (P<0.05) small bowel distension for three out of four segments (qualitative evaluation: two out of four segments) for methylcellulose in comparison to the control. Mannitol was found to be equal to the control. CONCLUSION: Oral ingestion of tap water or orange juice in combination with mannitol is recommended for cine MR imaging of the small bowel regarding luminal contrast and small bowel distension on bSSFP sequences. PMID- 17077039 TI - Magnetic resonance-guided biopsy of suspicious breast lesions with a handheld vacuum biopsy device. AB - PURPOSE: To evaluate a handheld vacuum-assisted device system for magnetic resonance image (MRI)-guided breast lesion biopsy. MATERIAL AND METHODS: In 32 patients, a total of 42 suspicious breast lesions (mean diameter 7.5 mm for mass lesions, 11.6 mm for non-masslike diffuse lesions) seen with MRI (no suspicious changes in breast ultrasound or mammography) were biopsied (27 lateral, 15 medial) using a 10G vacuum-assisted breast biopsy device under MR guidance. Histology of biopsy specimens was compared with final histology after surgery or follow-up in benign lesions. RESULTS: In all biopsies, technical success was achieved. Histology revealed 11 lesions with ductal carcinoma in situ (DCIS) or invasive cancer, three with intermediate lesions (LCIS) and 28 with benign breast lesions (adenosis, infected hematoma). In one patient with discordant results of MRI and histology, surgical excision revealed medullary cancer. In the follow-up (mean 18 months) of the histological benign lesions, no breast cancer development was observed. Besides minor complications (hematoma, n = 6), with no further therapeutic interventions, no complications occurred. CONCLUSION: MRI-guided breast lesion biopsy using a handheld vacuum-assisted device is a safe and effective method for the work-up of suspicious lesions seen with breast MRI without changes in mammography or ultrasound. In the case of discordant histology of vacuum biopsy and breast MRI appearance, surgical excision is recommended. PMID- 17077040 TI - Quantitative assessment of regional emphysema distribution in patients with chronic obstructive pulmonary disease (COPD). AB - PURPOSE: To compare objective and subjective assessment of the distribution of emphysema in unselected patients with chronic obstructive pulmonary disease (COPD). MATERIAL AND METHODS: 167 patients were computed tomography (CT) scanned, and the relative area (RA-910) of emphysema in each CT slice was plotted against table position. The craniocaudal distribution was calculated as the slope of the regression line, and grouped as upper-lung-zone predominance (ULP), lower-lung zone predominance (LLP), or mild/homogeneous distribution (MHE). CT scans were also classified as ULP, LLP, and MHE based on visual assessment of three high resolution CT (HRCT) slices, and the leading pattern of emphysema was classified as centrilobular (CLE), paraseptal (PSE), panlobular (PLE), or no emphysema (NE). RESULTS: By objective classification, scans were divided into almost equal numbers of ULP, LLP, and MHE, whereas visual evaluation classified more scans as ULP (P<0.001) and very few as LLP (P<0.0001). In patients with CLE, 49% had ULP by objective classification, whereas LLP was the commonest leading pattern in PSE, PLE, and NE. CONCLUSION: We found significant discrepancies between the objective and subjective distributions of emphysema in various morphological patterns, which may be of clinical importance in, for instance, lung-volume reduction surgery. PMID- 17077041 TI - Bronchial stump fistula: treatment with covered, retrievable, expandable, hinged stents--preliminary clinical experience. AB - PURPOSE: To evaluate the preliminary clinical efficacy and retrievability of a covered, retrievable, metallic hinged stent in the treatment of bronchial stump fistula (BPF). MATERIAL AND METHODS: Between April 2003 and March 2005, eight patients with bronchial stump fistula after pneumonectomy or lobectomy were treated with two types of covered, retrievable, metallic hinged stents. Type A stent was placed in six patients and type B in two under fluoroscopic guidance. The stent was removed with a retrieval set when BPF was healed or complications occurred. RESULTS: Stent placement in the bronchial tree was technically successful in all patients, without procedure-related complications. Immediate closure of the BPF was achieved in all patients after the procedure. Stents were removed from all patients but one. Removal of the stents was difficult in two patients due to tissue hyperplasia. Patients were followed for 4-16 months. Placement of the stents remained stable in all patients except one due to severe cough. Permanent closure of BPF was achieved in seven (87.5%) of eight patients. CONCLUSION: Use of a covered, retrievable, expandable, metallic hinged stent is a simple, safe, and effective procedure for the closure of bronchial stump fistula. Retrieval of the stent seems feasible. PMID- 17077042 TI - Scimitar syndrome versus meandering pulmonary vein: evaluation with three dimensional computed tomography. AB - We report two cases of abnormal configuration of the pulmonary venous system of the right lung evaluated with the use of three-dimensional spiral computed tomography (3D-CT). In the first case, an unusual form of scimitar syndrome was detected, consisting of two scimitar veins uniting into a common trunk immediately before their confluence into the inferior vena cava (IVC). The second case concerns a meandering pulmonary vein that followed a circuitous course through the right lung and came in close proximity to the IVC, before draining the entire right lung to the left atrium. In both cases, 3D-CT enabled a non invasive, detailed anatomic evaluation of the right pulmonary vascular bed. PMID- 17077044 TI - Pulmonary perfusion in acute pulmonary embolism: agreement of MRI and SPECT for lobar, segmental and subsegmental perfusion defects. AB - PURPOSE: To assess prospectively the agreement of magnetic resonance (MR) pulmonary perfusion with single-photon emission computed tomography (SPECT) perfusion for perfusion defects down to the subsegmental level in patients with suspected pulmonary embolism (PE). MATERIAL AND METHODS: In 41 patients with suspected PE, contrast-enhanced MR pulmonary perfusion (3D-FLASH, TR/TE 1.6/0.6 ms) was compared to SPECT perfusion on a per-examination basis as well as at the lobar, segmental, and subsegmental level. RESULTS: The MRI protocol was completed in all patients, and mean examination time was 3 min 56 s. MR perfusion showed a very high agreement with SPECT (kappa value per examination 0.98, and 0.98, 0.83, and 0.69 for lobar, segmental, and subsegmental perfusion defects, respectively). Of 15 patients with PE, MR perfusion detected 14 cases. CONCLUSION: The very high agreement of MR perfusion with SPECT perfusion enables the detection of subtle findings in suspected PE. PMID- 17077045 TI - Imaging of post-traumatic cardiac rhabdomyolysis with normal coronary arteries. AB - Numerous cases of acute myocardial infarction (AMI) have been reported in the literature following closed chest injuries, due to post-traumatic dissection or thrombosis of a coronary artery. In the follow-up of AMI, wall thickness during diastole and systole provides important information on heart viability. Multidetector computed tomography (MDCT) is currently the only noninvasive instrumental investigation which provides an appreciable assessment of the coronary arteries, as well as heart wall thickness measurements. We describe and discuss the clinical and imaging findings, especially of MDCT, in a case of post traumatic regional myocardial necrosis with normal coronary arteries. PMID- 17077046 TI - Bleeding pulmonary artery pseudoaneurysm secondary to squamous cell lung cancer: computed tomography findings and endovascular management. AB - A case of bleeding pulmonary artery pseudoaneurysm secondary to squamous cell lung cancer is reported. The patient presented with massive hemoptysis, diagnosis was made with multidetector computed tomography, and the pseudoaneurysm was successfully embolized with platinum coils. Hemoptysis ceased following the procedure. PMID- 17077047 TI - Low-field magnetic resonance imaging of the lumbar spine: reliability of qualitative evaluation of disc and muscle parameters. AB - PURPOSE: To determine the intra- and interobserver reliability in grading disc and muscle parameters using low-field magnetic resonance imaging (MRI). MATERIAL AND METHODS: MRI scans of 100 subjects representative of the general population were evaluated blindly by two radiologists. Criteria for grading lumbar discs were based on the spinal nomenclature of the Combined Task Force and the literature. Consensus in rating was achieved by evaluating 50 MRI examinations in tandem. The remaining 50 examinations were evaluated independently by the observers to determine interobserver agreement and re-evaluated by one of the observers to determine intra-observer agreement. RESULTS: Intra- and interobserver agreement was substantial when grading changes in the lumbar discs. Interobserver agreement was fair to moderate in grading the lumbar muscles, whereas intra-observer agreement was almost perfect. CONCLUSION: Convincing reliability was found in the evaluation of disc- and muscle-related MRI variables. PMID- 17077048 TI - Reproducibility of ultrasound and magnetic resonance imaging measurements of tendon size. AB - PURPOSE: To investigate the intra- and inter-tester reproducibility of measurements of the Achilles tendon, tibialis anterior tendon, and the tibialis posterior tendon in football players using ultrasound (US) and magnetic resonance imaging (MRI). MATERIAL AND METHODS: Eleven asymptomatic football players were examined. Using a standardized US scanning protocol, the tendons were examined by two observers with US for thickness, width, and cross-sectional area. One observer conducted the procedure twice. The subjects also underwent an MRI examination, and the assessment of tendon size was conducted twice by two observers. RESULTS: The best reproducibility judged by coefficient of variation (CV) and 95% confidence interval was determined for the Achilles tendon on both US and MRI. The variability of US on measurements on the tibialis anterior and tibialis posterior tendons was less than that when using MRI. In 12 out of 18 measurements, there were systematic differences between observers as judged by one-sided F-test. CONCLUSION: The reproducibility of the three tendons was limited. Precaution should be taken when looking for minor quantitative changes, i.e., training-induced hypertrophy, and when doing so, the Achilles tendon should be used. PMID- 17077049 TI - Increasing benefit of magnetic resonance imaging in multiple sclerosis. AB - Magnetic resonance imaging (MRI) has emerged as an essential tool of multiple sclerosis (MS) diagnosis and has opened up completely new prospects in MS research and treatment trials. It is a sensitive method that gives direct evidence of tissue pathology and has greatly increased our knowledge of MS. In clinical work, MRI is used to confirm and exclude the diagnosis of MS. The international recommendation is that every suspected MS patient should undergo at least one brain MRI. T2-weighted images are the standard tool in clinical work, and functional imaging methods are mainly used in MS research. The subtypes and the course of the disease cause variation in MRI findings. Here, we present a general overview of MR findings in MS. PMID- 17077050 TI - Diffusion tensor imaging and magnetic resonance spectroscopy of the brain in a patient with Sturge-Weber syndrome. AB - Quantitative brain MR spectroscopy (MRS) and diffusion tensor imaging (DTI) were used to characterize one patient with Sturge-Weber syndrome. Choline increases and N-acetylaspartate decreases were observed in pathologic frontal gray matter tissue compared to contralateral unaffected brain tissue without any change in the diffusion tensor imaging parameters (fractional anisotropy, apparent diffusion coefficient). The N-acetylaspartate decreases and/or choline increases observed here and in eight previously described Sturge-Weber patients probably reflect neuronal loss or dysfunction and demyelination as a result of recurrent seizures. PMID- 17077051 TI - Can dynamic susceptibility contrast magnetic resonance imaging replace single photon emission computed tomography in the diagnosis of patients with Alzheimer's disease? A pilot study. AB - PURPOSE: To compare single-photon emission computed tomography (SPECT) and magnetic resonance imaging (MRI) in a cohort of patients examined for suspected dementia, including patients with no objective cognitive impairment (control group), mild cognitive impairment (MCI), and Alzheimer's disease (AD). MATERIAL AND METHODS: Twenty-four patients, eight with AD, 10 with MCI, and six controls were investigated with SPECT using 99mTc-hexamethylpropyleneamine oxime (HMPAO) and dynamic susceptibility contrast magnetic resonance imaging (DSC-MRI) with gadobutrol. Three observers performed a visual interpretation of the SPECT and MR images using a four-point visual scale. RESULTS: SPECT was superior to DSC-MRI in differentiating normal from pathological. All three observers showed statistically significant results in discriminating between the control group, AD, and MCI by SPECT, with a P value of 0.0006, 0.04, and 0.01 for each observer. The statistical results were not significant for MR (P values 0.8, 0.1, and 0.2, respectively). CONCLUSION: DSC-MRI could not replace SPECT in the diagnosis of patients with Alzheimer's disease. Several patient- and method-related improvements should be made before this method can be recommended for clinical practice. PMID- 17077052 TI - Magnetic resonance imaging of the ovaries of healthy women: determination of normal values. AB - PURPOSE: To establish normal values for the volume and maximal diameter of ovaries and ovarian follicles and for the number of ovarian follicles in magnetic resonance imaging (MRI) based on menstrual cycle phase and age. MATERIAL AND METHODS: We performed MRI of the pelvis on 100 healthy women. Volume of the ovaries and largest ovarian follicles and the number of ovarian follicles were determined by menstrual cycle phase and age. RESULTS: The mean volume of the ovaries significantly increased with age and reached its peak between 31 and 40 years, and subsequently decreased. The mean volume of the largest ovarian follicles also significantly increased with age to reach its peak at 41-50 years. The highest mean numbers of ovarian follicles were found at 20-40 years. When the volumes of ovaries and of the largest ovarian follicles, and the number of ovarian follicles were compared between the first and second phase of the menstrual cycle, no significant differences were found. CONCLUSION: The volume and maximal diameter of ovaries and ovarian follicles and the number of ovarian follicles differ significantly with age, but not between the two phases of the menstrual cycle. Knowledge of MRI-related normal values can be expected to aid the early identification of ovarian pathologies. PMID- 17077053 TI - Imaging of primary adrenal lymphoma: case report and literature review. AB - Primary adrenal lymphoma is very rare, and correct preoperative diagnosis is difficult because of its nonspecific imaging findings. We report a case of bilateral primary adrenal lymphoma, with the largest dimension of 18 cm, which is the largest one reported so far, to our knowledge. Computed tomography (CT) and magnetic resonance (MR) findings were not specific enough to establish a correct diagnosis, but positron emission tomography/CT (PET/CT) showed high fluorodeoxyglucose (FDG) uptake, indicating its malignant nature. We describe the imaging findings and review the related literature to highlight this uncommon entity. PMID- 17077054 TI - Myomatous erythrocytosis syndrome: a case report. AB - We report the case of a 36-year-old woman with erythrocytosis due to ectopic erythropoietin production by a very large uterine leiomyoma. Awareness of this uncommon condition is important so that the correct diagnosis can be suggested prior to surgery and radical resection can be avoided. PMID- 17077055 TI - Pseudoaneurysm of the renal interlobar artery in Behcet's disease. AB - Behcet's disease is a rare clinical event characterized by recurrent oral and genital ulcers, and iridocyclitis. Vascular involvement is a rare but serious form of the disease. We present a case of renal interlobar artery pseudoaneurysm occurring in a 20-year-old male patient with a 5-year history of Behcet's disease, who had been admitted because of right flank pain. Findings obtained on abdominal ultrasonography, abdominal computed tomography, and renal angiography confirmed a right renal pseudoaneurysm and subcapsular hematoma. PMID- 17077056 TI - Data analysis in radiology research. PMID- 17077058 TI - High protein high fibre snack bars reduce food intake and improve short term glucose and insulin profiles compared with high fat snack bars. AB - The replacement in the diet of refined carbohydrate and fat with fibre and protein has been shown to promote satiety and improve glucose and insulin profiles. It is less clear whether the macronutrient composition of individual foods such as snacks have any meaningful impact on metabolic parameters and satiety. We examined if the consumption of higher protein higher fibre snack bars would result in reducing outcome measures such as food intake and glucose and insulin patterns compared to a conventional isocaloric high fat high refined carbohydrate snack bar. Twenty three women were randomized in a single blind cross over study with 2 interventions, a high fat high sugar snack bar and a comparatively higher protein, higher fibre snack bar intervention. Snack bars were eaten at mid morning and mid afternoon, and a standard breakfast and ad libitum buffet lunch. The glucose and insulin responses over 9 hours were significantly lower (P = 0.014 and P = 0.012 respectively) during the high protein snack bar intervention. Peak glucose levels were also 16% lower after the morning HP bar (P <0.001). The morning high protein bar reduced the energy intake at the buffet lunch meal by 5% (4657 +/- 1025KJ vs 4901 +/- 1186KJ, P < 0.05). Altering the macronutrient composition of a snack bar can assist in reducing the energy intake at a subsequent meal and improve short term glucose and insulin profiles. PMID- 17077059 TI - Precision in nutritional information declarations on food labels in Australia. AB - Nutrition labels guide consumers in making their food choice. New requirements for mandatory nutrition labelling have been in force in Australia since late 2002. The present study, analysing 350 samples comprising 70 different products for nutritional compounds declared on the label, is the first larger attempt to quantify the precision in nutrition labelling of food products on the Australian market. A significant discrepancy between actual and declared values was detected with an average variation in precision of -13% to +61% for individual nutritional components. There is no tolerance limit established in the Australian food legislation but a +/- 20% discrepancy is allowed in some countries and others have separate upper and lower limits and allow a maximum discrepancy of -;20% for beneficial nutritional compounds and +20% for unfavourable compounds. Only 16% of the 70 products in the study would fully comply should a leeway of +/-20% be introduced for any nutritional compound on the label. With separate upper and lower limits, 51% of products would fully comply. Compliance improved to 27% and 70% of products, respectively, when excluding variations in minor amounts irrelevant to consumers (counting all variations of less than 1g/100g, or 10kJ/100g for energy and 10mg/100g for sodium, potassium, calcium and cholesterol, as compliant). It is proposed that adoption of an upper and lower tolerance limit, excluding minor amounts, be considered as a way of better assisting the consumer in making relevant comparisons of product nutritional value and that any nutritional component should comply with the limit for the product to comply. Applying such a limit, 30% of products in the survey would not be compliant. PMID- 17077060 TI - Under-reporting of energy intake affects estimates of nutrient intakes. AB - Under-reporting of energy intake is a common problem in nutritional epidemiological studies. The aim of the present study was to determine the effect of under-reporting of energy intake on the estimates of nutrient intakes. In this cross-sectional study, 901 subjects aged >16 y were randomly selected from participants of the Tehran Lipid and Glucose Study. Dietary intake was assessed by means of two 24-hour dietary recalls. Basal metabolic rate (BMR) was determined according to age, sex and weight. The ratio of energy intake (EI) to BMR was calculated. Under-reporting of energy intake was defined as EI:BMR< 1.35 and normal-reporting of dietary intake as EI:BMR = 1.35-2.39. To obtain energy adjusted amounts of macro- and micronutrients, the residual method was used. Under-reporting of energy intake was revealed in 31% of the subjects and was more common among females (40%) than males (19%, P <0.01). The mean age of females who under-reported was significantly lower than the normal-reporting females (32 +/- 13 vs. 35 +/- 14 y, P <0.05); however, the age difference between the two groups was not significant in men. Under-reporters had higher BMIs compared to normal reporters in both genders. The absolute intakes of macro- and micronutrients (except for B12 in females and B6 and zinc in both genders) were lower in under reporters, but following adjustment, no significant differences were seen. The results have revealed that under-reporting of energy intake affects the estimates of nutrient intakes; thus in studies aimed at determining the association between a certain chronic disease and a nutrient intake, we suggest adjustments be made for energy intake. PMID- 17077061 TI - Calibration and reliability of a school food checklist: a new tool for assessing school food and beverage consumption. AB - There is a pressing need in Australia and other countries to develop systems for monitoring secular trends in childhood obesity and related behavioural and environmental determinants. Energy from foods and beverages consumed at school is an accessible indicator of children's eating patterns and we have developed a school food checklist (SFC) to measure this. The SFC records the number of serves and source (home, canteen, vending machine) of 20 food and beverage categories. This study aims to assess the accuracy and to calibrate the SFC by comparing it to a weighed record (WR) and to evaluate inter-recorder reliability. Participants were 910 primary school children aged 5 to 12 years from a rural township in Victoria, Australia. WR were collected from a non-random sub-sample of 106 and a second sub-sample (n=46) had intake measured twice using the SFC to assess inter recorder reliability. Mean energy values were 2992 kJ +/- 924 and 3008 kJ +/- 952 for the SFC and WR respectively and the correlation coefficient was strong (Pearson r = 0.77). The mean difference between the WR and SFC methods was 15 kJ (95% CI, -107 kJ to 138 kJ) and the limits of agreement (+2 standard deviations) were +/- 1270 kJ. The SFC overestimated the energy/serve of breads and fruit drinks and under-estimated energy/serve from fat spreads, biscuits/crackers, muesli/fruit bars and fruit. Inter-recorder reliability was good (kappa 0.51). The SFC was designed to measure energy from food and beverages in schools. It has good accuracy and reliability and the revised version should further improve accuracy of the instrument. PMID- 17077062 TI - Primary school children from northeast Thailand are not at risk of selenium deficiency. AB - Selenium has important roles as an antioxidant, in thyroid hormone metabolism, redox reactions, reproduction and immune function, but information on the selenium status of Thai children is limited. We have assessed the selenium status of 515 northeast Thai children (259 males; 256 females) aged 6 to 13 years from 10 rural schools in Ubon Ratchthani province. Serum selenium (n=515) was analyzed by Graphite Furnace Atomic Absorption Spectrophotometry and dietary selenium intake by Hydride Generation Absorption Spectrophotometry from one-day duplicate diet composites, from 80 (40 females; 40 males) randomly selected children. Inter relationships between serum selenium and selenium intakes, and other biochemical micronutrient indices were also examined. Mean (SD) serum selenium was 1.46 (0.24) micro mol/L. Concentrations were not affected by infection or haemoglobinopathies, but were dependent on school (P< 0.001), sex (P=0.038), and age group (P=0.003), with serum zinc as a significant covariate. None of the children had serum selenium concentrations indicative of clinical selenium deficiency (i.e. <0.1 micro mol/L). Significant correlations existed between serum selenium and serum zinc (r= 0.216; P < 0.001), serum retinol (r = 0.273; P < 0.001), urinary iodine (r = -0.110; P = 0.014), haemoglobin (r = 0.298; P <0.001), and haematocrit (r = 0.303; P< 0.001). Mean (SD) dietary selenium intake was 46 (22) micro g/d. Children with low serum selenium concentrations had a lower mean selenium intake than those with high serum selenium concentrations (38 +/- 17 vs.51 +/- 24 micro g/d; P< 0.010). In conclusion, there appears to be no risk of selenium deficiency among these northeast Thai children. PMID- 17077063 TI - Obesity and undernutrition in sub-Saharan African immigrant and refugee children in Victoria, Australia. AB - The study assessed the anthropometric status of 337 sub-Saharan African children aged between 3-12 years who migrated to Australia. These children were selected using a snowball sampling method stratified by age, gender and region of origin. The prevalence rates for overweight and obesity were 18.4% (95%CI: 14 - 23%) and 8.6% (95%CI: 6% -12%) respectively. The prevalence rates for the indicators of undernutrition were: wasting 4.3% (95%CI: 1.6%-9.1%), underweight 1.2% (95%CI: 0.3%-3.0%), and stunting 0.3 (95%CI: 0.0%-1.6%). Higher prevalence of overweight/obesity was associated with lower household income level, fewer siblings, lower birth weight, western African background, and single parent households (after controlling for demographic and socio-economic factors). Higher prevalence rates for underweight and wasting were associated with lower household income and shorter lengths of stay in Australia respectively. No effect was found for child's age, gender, parental education and occupation for both obesity and undernutrition indices. In conclusion, obesity and overweight are very prevalent in SSA migrant children and undernutrition, especially wasting, was also not uncommon in this target group. PMID- 17077064 TI - Risk factors for malnutrition among rural Nigerian children. AB - Protein Energy Malnutrition (PEM) remains a major public health problem in the developing world. The aim of this study was to determine the current nutritional status and the influence of feeding practices and family characteristics on the nutritional status of under-five rural Nigerian children. It was conducted using a cross sectional, community based survey design. From 344 households, 420 children were studied. Using the modified Wellcome Classification, the prevalence of PEM was 20.5 percent whereas the prevalence of underweight, wasting and stunting using the World Health Organization/ National Centre for Health Statistics (WHO/NCHS) standards were 23.1 percent, 9 percent and 26.7 percent respectively. Young age was significantly associated with a higher prevalence of underweight (P = 0.004). Overcrowding, low maternal income and the use of infant formula feeds in children who have attained the age of 6 months and above were associated with a higher prevalence of wasting (P = 0.029, P = 0.031 and P = 0.005 respectively). Improved living standard of families, empowerment of mothers with the aim of augmenting family income and parental education on appropriate feeding practices may help in reducing the incidence of under-five malnutrition in communities. The low prevalence rate of malnutrition was probably due to activities of the NGO in this community. This method of intervention is similarly achievable in any other community. PMID- 17077065 TI - Low density lipoprotein subclasses in Asian and Caucasian adolescent boys. AB - South Asian adults are known to have very high rates of Coronary heart disease (CHD) and insulin resistance and, even as adolescents, may show higher risk factors for CHD. The aim of this study was to investigate the prevalence of small, dense low density lipoprotein (sdLDL) subclasses in a cohort of adolescent boys. The specific objective was to investigate the relationship between measures of fatness, ethnicity and LDL diameter in this cohort. Preformed native (non denaturing) polyacrylamide 3-13% gradient gels and a multipurpose vertical electrophoresis system were used for the separation of LDL sub-fractions in a single school year cohort of boys aged 15-16 years (n=135). Latex beads and thyroglobulin standards were used to construct a calibration curve in order to calculate LDL particle diameters by regression (Total Lab Software v1.11). ANOVA was used to compare LDL size among different ethnic groups (SPSS and Stat View). The study sample was comprised of 45.2% Caucasians, 41.5% East Asians and 13.3% from the Indian subcontinent (South Asians). There was a non-significant trend for South Asians to have a lower LDL diameter than either Caucasians or East Asian boys which was independent of % total body fat (%TBF) and body mass index (BMI). This is the first adolescent cohort to examine sdLDL which included Caucasians, East and South Asians. It appears that the higher risk profile for CHD and diabetes noted in South Asian adults may be evident even during adolescence. PMID- 17077066 TI - Is the BMI cut-off level for Japanese females for obesity set too high? A consideration from a body composition perspective. AB - The aim of the study was to determine the sensitivity and specificity of the BMI cut-off points that have been applied in the National Nutritional Survey in Japan (NNS-J) for young Japanese females. The relationship between the body mass index (BMI) and percentage body fat (%BF) of 139 young Japanese females (aged 18-27 years old) was examined using dual energy x-ray absorptiometry (DXA) and anthropometry. The sensitivity and specificity of the BMI cut-off point that has been in use in Japan was compared with the cut-off point proposed for Asia Pacific population (the WPRO classification). The regression equation calculated for the relationship between the BMI and %BF using the DXA scan data was %BF = 15.152+2.058*BMI; R2 = 0.612, SEE = 3.66. From this equation the BMI value that corresponded with %BF of 30% was calculated to be 21.9kg/m2 (95%CI: 16.9-28.6). From the sensitivity and specificity analysis, it was found that more than 90% of Japanese females with %BF >30% were misclassified as "not obese" when the BMI value of 25kg/m2 was used to identify obese individuals. The misclassification reduced to 60% when the BMI value of 23kg/m2 was used as a cut-off point. The results indicate that the current classification used in the NNS-J may not precisely reflect the obesity status of young Japanese females and a considerable proportion of females with a large fat deposition would be misclassified as not obese. The current study shows the importance of including detailed body composition assessments to determine obesity level of individuals. PMID- 17077067 TI - The relationship between obesity and health related quality of life of women in a Turkish city with a high prevalence of obesity. AB - The purpose of this study was to demonstrate the relationship between body weight and HRQOL in a representative sample of nonpregnant women in reproductive age period. The data of this cross-sectional study was extracted from a survey: Manisa Demographic and Health Survey (MDHS) conducted in Manisa city in 2000. The study population of MDHS is a representative sample of 1602 reproductive (15-49) age women. World Health Organization Quality of Life Questionnaire abbreviated version (WHOQOL-BREF), which was composed of four domain factors (physical, psychological, social relations and the environment), was used to assess HRQOL. Each of four domains had a possible score ranged between 0 (poor HRQOL) and 20 (excellent HRQOL). The mean age of the women was 35.29+/- 8.19 years. Among them, 35.8 % had normal weight (BMI 18.5 to 24.9), 32.3 % were overweight (BMI 25.0 to 29.9) while 31.9 % were moderate and 3.4% were morbidly obese. After adjusting for age, level of education and co-morbid illnesses, subjects with a BMI higher than normal value, had significantly lower HRQOL scores, compared to normal weight individuals on each of the domains, except for the environmental domain. Our results suggested that the body weight alone could negatively affect HRQOL. In other words, obesity not only increased the risk of morbidity and mortality, but also affected the perceived health and life quality negatively. In conclusion, in addition to age, socioeconomic status and co-morbid illnesses, body weight should also be controlled in studies examining HRQOL. PMID- 17077068 TI - Obesity among university students, Tehran, Iran. AB - Evaluating the nutritional status of individuals and population groups is an important tool in public health and a feasible indicator of standards of living. The objective of this study was to determine the frequency of obesity and present nutritional status of university students of Iran University of Medical Sciences in Tehran. The survey was conducted between Oct 2004 and June 2005. The statistical population included all students from School of medicine, nursing and midwifery, health services, management, science, and rehabilitation. The method of sampling was multistage random. The sample size for the survey was 1,150 students. We used a self-administered 24h food recall questionnaire. We categorized BMI to two groups in bivariate analysis (BMI <25 kg m(-2) as obese and less than 25 kg m(-2) as non-obese). Mean BMI for all subjects was 21.7 +/- 2.9 kg m(-2). Almost 88% of the subjects were classified into a non-obese group (BMI <25 kg m(-2)). About 10% were underweight and 12.4% of the students had a BMI more than 25 kg m(-2). A significant difference was observed for BMI between males and females; 7.9% of males versus 22.5% of females had BMIs over 25. About 18% of students aged 23 years and over had BMIs over 25 versus 7.7% of students aged under 18. Intakes of fiber, pre-vitamin A, folacin and iron were significantly different between BMI groups. Intakes of these nutrients were higher in the obese students than the students with BMIs less than 25 kg m(-2). Our results indicate that about 12.4% of the students had a BMI more than 25 kg m(-2). There was a significant association between BMI, and smoking habits, age, sex, place of resident and having specific dietary regimen. PMID- 17077069 TI - Alternative anthropometric measurements for the Thai elderly: Mindex and Demiquet. AB - This cross-sectional study examined the relationship between alternative anthropometric indices and the nutritional and metabolic status of the Thai elderly. Four rural communities, each from the 4 main regions of Thailand were surveyed. A total of 2,324 subjects, 60 years old and over were included in the study. Mindex and Demiquet had a very strong relationship to body mass index with the r values of 0.93 and 0.88, respectively. Fat weight had the highest correlation with body mass index in older women, r = 0.94 (P< 0.001) and with Mindex, r = 0.93 (P< 0.001). In regard to anthropometric measurements, the mid arm circumference had the strongest relationship to all three Quetelet indices, r = 0.76-0.87 (P< 0.001), while subscapular skinfold thickness performed best among skinfold measurements, r = 0.69-0.74 (P< 0.001). BMI achieved a significantly higher degree of correlation with triceps and supra-iliac skinfold thickness, mid arm circumference and fat weight than Demiquet (P< 0.001 for all variables). The lymphocyte count was the only laboratory test that related rather well to all three Quetelet indices. All three Quetelet indices had nearly the same pattern of relationship to various nutritional parameters. The cut-off points of Mindex denoting under-nutrition, overweight and obesity I in women were 55.95, 69.55 and 75.60 kilogram/metre, respectively. At the same time, the cut-off points of Demiquet denoting under-nutrition, overweight and obesity I in men were 75.60, 93.98 and 102.16 kilogram/metre2, respectively. All this information supports the benefit of using Mindex and Demiquet as alternatives to body mass index for nutritional assessment in older Asian people, especially for the malnourished ones. PMID- 17077070 TI - Persistence of iodine deficiency in Gangetic flood-prone area, West Bengal, India. AB - In 2000, India revoked the ban on production and sale of non-iodised salt. We conducted a study in the north 24 Parganas district in the state of West Bengal to assess the prevalence of goitre, status of urinary iodine excretion (UIE) level and to estimate iodine content of salts at the household level. We surveyed 363 school children aged eight to ten years selected using a multistage cluster sampling technique. We estimated goitre prevalence and urinary iodine excretion (UIE) using methods and criteria recommended by the World Health Organization. We estimated the iodine content of salt samples collected from the households of the study subjects using spot iodine testing kit. Of the 363 children, 73 (20%) had goitre. The median UIE was 160 micro g/l (normal: > or =100 micro g/l) and only 6% children had a level below 50 micro g/l. Only 253 of 363 salt samples (70%) were sufficiently iodised. The combination of high goitre prevalence with normal median urinary excretion indicates that the North 24 Parganas district is in transition from iodine deficient to iodine sufficient state. However, the persistence of non-iodised salt consumption indicates that an intensification of universal salt iodisation program is needed. In 2000, India revoked the ban on production and sale of non-iodised salt. We conducted a study in the north 24 Parganas district in the state of West Bengal to assess the prevalence of goitre, status of urinary iodine excretion (UIE) level and to estimate iodine content of salts at the household level. We surveyed 363 school children aged eight to ten years selected using a multistage cluster sampling technique. We estimated goitre prevalence and urinary iodine excretion (UIE) using methods and criteria recommended by the World Health Organization. We estimated the iodine content of salt samples collected from the households of the study subjects using spot iodine testing kit. Of the 363 children, 73 (20%) had goitre. The median UIE was 160 micro g/l (normal: > or =100 micro g/l) and only 6% children had a level below 50 micro g/l. Only 253 of 363 salt samples (70%) were sufficiently iodised. The combination of high goitre prevalence with normal median urinary excretion indicates that the North 24 Parganas district is in transition from iodine deficient to iodine sufficient state. However, the persistence of non-iodised salt consumption indicates that an intensification of universal salt iodisation program is needed. PMID- 17077071 TI - Status of iodine nutrition in pregnant and lactating women in national capital district, Papua New Guinea. AB - Urinary Iodine excretion is a useful and important indicator of the iodine status of a population. This study attempts to determine the urinary iodine concentration of non-pregnant, pregnant and lactating women, resident in the National Capital District of Papua New Guinea, so as to evaluate their status of iodine nutrition. The study population was made up of 56 non-pregnant, 40 lactating and 212 pregnant women. Of the 212 pregnant women, 14 were in the first, 64 in the second, and 134 in the third Trimester of pregnancy. Casual urine samples were collected and analysed for urinary iodine by Sandell-Kolthoff reaction. The median urinary iodine concentration for the non-pregnant, lactating and pregnant women was 163.0 micro g/L, 134.0 micro g/L and 180.0 micro g/L, respectively. Median urinary iodine for the first, second and third trimesters were 165.0 micro g/L, 221.5 micro g/L and 178.0 micro g/L, respectively. The 20th percentile urinary iodine values were higher than 50 micro g/L for all the groups. This indicates adequate intake of dietary iodine and optimal status of iodine nutrition amongst women in the various groups. Mild to severe status of iodine nutrition was found in 30.4% of non-pregnant, 35.0% of lactating, 22.2% of pregnant women, 28.5% of women in the first, 18.8% in the second, and 23.1% in the third trimester of pregnancy. To achieve optimal iodine nutrition in pregnant and lactating women, an increase in their intake of dietary iodine is recommended. PMID- 17077072 TI - The relationship of neonatal serum vitamin B12 status with birth weight. AB - Earlier studies have shown a relationship between maternal vitamin B12 status and birth weight. This study extends those findings directly in terms of neonatal vitamin B12 status and birth weight. One hundred and twelve women were followed from the first trimester of pregnancy and maternal blood was obtained in all three trimesters along with cord blood at birth of their neonates. The maternal and cord serum vitamin B12 concentrations were examined in relation to birth weight. There was a significant correlation between vitamin B12 concentration in maternal antenatal serum during each of the trimesters of pregnancy and cord serum (all P< 0.01). Neonates that were born with lower birth weights (categories of <2500 g and 2500-2999g) had significantly lower mean cord serum vitamin B12 concentrations when compared to those who were > or = 3000g (P = 0.02 and P = 0.05 respectively). A similar, however, non significant trend was observed for antenatal vitamin B12 concentrations at first and third trimesters. Cord serum vitamin B12 concentrations were significantly correlated with birth weight, up to 40 weeks of pregnancy (r=0.28, P=0.01) but not beyond that (> or =40 weeks gestation). Vitamin B12 status in the mother was related to neonatal vitamin B12 status as measured by cord serum vitamin B12 concentration. In addition, low neonatal vitamin B12 concentrations were adversely associated with low birth weights. PMID- 17077073 TI - Anti-inflammatory and anticatabolic effects of short-term beta-hydroxy-beta methylbutyrate supplementation on chronic obstructive pulmonary disease patients in intensive care unit. AB - Elevated inflammatory markers and muscle wasting were common in chronic obstructive pulmonary disease (COPD) patients. The purpose of this study was to investigate the effect of 7-day beta-hydroxy-beta-methylbutyrate (HMB) supplementation on inflammation, protein metabolism, and pulmonary function in COPD patients in an intensive care unit. Thirty-four COPD patients who required mechanical ventilators were randomly assigned to HMB (n=18) or control (n=16) groups. The HMB group received HMB 3 g/d for 7 days. White blood cell count, C reactive protein, and creatinine were significantly lower, while cholesterol and total protein were significantly higher after HMB supplementation. The body weight remained unchanged in both groups. Ten subjects (55.6%) in the HMB group and 4 subjects (25.0%) in the control group had improved pulmonary function, indicated by their ventilator modes. This short-term study suggests that HMB supplementation may have anti-inflammatory and anticatabolic effect and improve pulmonary function in COPD patients in an intensive care unit setting. PMID- 17077074 TI - Effects of dietary supplements on the Fischer ratio before and after pulmonary rehabilitation. AB - Recently, efforts in comprehensive pulmonary rehabilitation for COPD have been made, including education, physical therapy, occupational therapy, nutrition, nursing, medication and counseling. Each patient focuses on a different element. Supplying adequate nutrition, among others, is essential for comprehensive pulmonary rehabilitation, as well as survival. In this study, the utility of efficient nutritional supplement therapy before and after pulmonary physical therapy was investigated by adding an amino acid drink with a high Fisher ratio to comprehensive pulmonary rehabilitation. The subjects were eight patients with COPD with obstructive ventilation disorder as severe as 31.5 +/- 6% of FEV 1.0%. Pulmonary physical therapy was performed for eight weeks in a group administered one bottle of dietary supplement with a high Fisher ratio abundant in branched chain amino acids once daily (200 kCal/ 200 mL, Fisher ratio 40), and in another group without administration. Before and after the physical therapy, six-minute waking examination, QOL assessment (using CRQ), serum protein and serum Fisher ratio were comparatively examined between the two groups. After the eight weeks of pulmonary physical therapy, serum Fisher ratios were evidently reduced and serum protein measurements were also decreased in the group without dietary supplement abundant in branched chain amino acids. Accordingly, more amino acid is needed due to enhanced consumption of muscular protein during pulmonary physical therapy, during which nutrient ingestion including a sufficient amount of branched amino acid is necessary. It is an important element in continuing comprehensive pulmonary rehabilitation for a longer period. PMID- 17077075 TI - Influencing health through intestinal microbiota modulation and probiotics. AB - Science has progressed fast in providing answers to probiotic health benefits to the consumers. This symposium also proves that progress is fast. However, several challenges still need to be solved and more effective strains and strain combinations discovered. This will pave the way from good probiotics to specific products for clearly identified target populations. The intestinal microbiota and its interaction(s) with probiotics challenges researchers to turn to the next new page to discover new approaches and treatment modalities that utilize probiotics as means of providing good nutrition with clear health benefits to all consumers. PMID- 17077076 TI - Intestinal colonisation, microbiota and future probiotics? AB - The human intestine is colonized by a large number of microorganisms, collectively termed microbiota, which support a variety of physiological functions. As the major part of the microbiota has not yet been cultured, molecular methods are required to determine microbial composition and the impact of specific dietary components including probiotics. Probiotics are viable microbial food supplements, which have a beneficial impact on human health. Health-promoting properties have been demonstrated for specific probiotic products. The most significant demonstrations for probiotic efficacy include prevention and treatment of antibiotic associated diarrhea, rotavirus diarrhea and allergy prevention. Lactobacillus rhamnosus GG (=ATCC 53103) and Bifidobacterium lactis Bb12 are the among the best-characterized and most studied probiotic strains with demonstrated impact on human health. New complex targets for probiotics include irritable bowel syndrome and Helicobacter pylori infection. For future probiotics the most important target is a demonstrated clinical benefit supported by knowledge on the mechanistic actions in the microbiota of the target population. Molecular and genomics-based knowledge of the composition and functions of the microbiota, as well as deviations from the balanced microbiota, will advance the selection of new and specific probiotics. Potential combinations of specific probiotics may prove to be the next step to reduce the risk on intestinal diseases and reconstruct specific microbial deviations. PMID- 17077077 TI - Safety of probiotics. AB - New species and more specific strains of probiotic bacteria are constantly being sought for novel probiotic products. Prior to the incorporation of novel strains into food or therapeutic products a careful evaluation of their efficacy is required and an assessment made as to whether they share the safety status of traditional food organisms. Food organisms intrinsic to the production of traditional foods have been arbitrarily classified as safe in the absence of scientific criteria. Evidence for the safety and efficacy of probiotics has until recently been largely anecdotal or based on relatively little, and often poorly designed research. The demonstration of efficacy in probiotics offers vast opportunities for the development of human and veterinary products. The introduction of a new probiotic culture demands that it be at least as safe as its conventional counterparts. Many bacteria are being tested to find a putative probiotic, yielding conflicting data, sometimes for the same organism. Comparisons between studies and organisms cannot be readily made because of non standardized dosing procedures. Information is not readily available on the equivalence of formulations for different probiotic preparations. There is vigorous debate on what constitutes appropriate safety testing for novel probiotic strains proposed for human consumption. Conventional toxicology and safety evaluation is of limited value in assessing the safety of probiotics. The addition of novel bacterial strains to foods and therapeutic products requires reconsideration of the procedures for safety assessment. This paper provides an overview of these issues. PMID- 17077078 TI - Protection mechanism of probiotic combination against human pathogens: in vitro adhesion to human intestinal mucus. AB - In this study we evaluated the ability of commercial strains (L. rhamnosus GG, L. rhamnosus LC705, and P. freudenreichii ssp. shermanii JS) in combination with B. breve 99 or B. lactis Bb12 to inhibit, displace and compete with model pathogens in order to test their influence on the adhesion of selected pathogens to immobilized human intestinal mucus. Our results demonstrate that specific probiotic combinations are able to enhance the inhibition percentages of pathogens adhesion to intestinal mucus when compared to individual strains. This suggests that combinations of probiotic strains are useful and more effective in inhibition of pathogen adhesion than individual strains. Such combinations should be assessed in clinical studies in subjects where the intestinal microbiota aberrancies have been identified. PMID- 17077079 TI - Clinical studies on alleviating the symptoms of irritable bowel syndrome. AB - Irritable bowel syndrome (IBS) is one of the most common diagnoses in gastroenterology, but current therapies are inefficient. Recent clinical trials suggest beneficial effects of certain probiotics in IBS. Because of the heterogeneity of IBS a probiotic combination may be more efficient than a single strain. We screened for optimal strains, and developed a multispecies probiotic combination consisting of L. rhamnosus GG, L. rhamnosus Lc705, P. freudenreichii ssp. shermanii JS and Bifidobacterium breve Bb99. The clinical efficacy of the probiotic combination was evaluated in IBS patients in a randomised, double blind, placebo-controlled six-month intervention. During six months the subjects received daily either probiotic supplementation or placebo. IBS symptoms were followed by symptom diaries. The probiotic supplementation demonstrated significant value in reducing IBS symptoms. At the end of the study period the total symptom score (abdominal pain + distension + flatulence + rumbling) had reduced with 42% in probiotic group versus 6% for instance anti-inflammatory effects, balancing of the microbiota or motility-related effects induced by the probiotic. The probiotic activity may be enhanced by synergistic effects of the combination that each strain alone would not hold. In conclusion, we found a probiotic combination of LGG and three other strains to be effective in alleviating IBS symptoms. PMID- 17077080 TI - Stress-induced inactivation of the c-Myb transcription factor through conjugation of SUMO-2/3 proteins. AB - Post-translational modifications, such as phosphorylation, acetylation, ubiquitination, and SUMOylation, play an important role in regulation of the stability and the transcriptional activity of c-Myb. Conjugation of small ubiquitin-like modifier type 1 (SUMO-1) to lysines in the negative regulatory domain strongly suppresses its transcriptional activity. Here we report conjugation of two other members of the SUMO protein family, SUMO-2 and SUMO-3, and provide evidence that this post-translational modification negatively affects transcriptional activity of c-Myb. Conjugation of SUMO-2/3 proteins is strongly enhanced by several different cellular stresses and occurs primarily on two lysines, Lys(523) and Lys(499). These lysines are in the negative regulatory domain of c-Myb and also serve as acceptor sites for SUMO-1. Stress-induced SUMO 2/3 conjugation is very rapid and independent of activation of stress-activated protein kinases of the SAPK and JNK families. PIAS-3 protein was identified as a new c-Myb-specific SUMO-E3 ligase that both catalyzes conjugation of SUMO-2/3 proteins to c-Myb and exerts a negative effect on c-Myb-induced reporter gene activation. Interestingly, co-expression of a SPRING finger mutant of PIAS-3 significantly suppresses SUMOylation of c-Myb under stress. These results argue that PIAS-3 SUMO-E3 ligase plays a critical role in stress-induced conjugation of SUMO-2/3 to c-Myb. We also detected stress-induced conjugation of SUMO-2/3 to c Myb in hematopoietic cells at the levels of endogenously expressed proteins. Furthermore, according to the negative role of SUMO conjugation on c-Myb capacity, we have observed rapid stress-induced down-regulation of the targets genes c-myc and bcl-2 of c-Myb. Our findings demonstrate that SUMO-2/3 proteins conjugate to c-Myb and negatively regulate its activity in cells under stress. PMID- 17077081 TI - Altered -3 substrate specificity of Escherichia coli signal peptidase 1 mutants as revealed by screening a combinatorial peptide library. AB - Signal peptidase functions to cleave signal peptides from preproteins at the cell membrane. It has a substrate specificity for small uncharged residues at -1 (P1) and aliphatic residues at the -3 (P3) position. Previously, we have reported that certain alterations of the Ile-144 and Ile-86 residues in Escherichia coli signal peptidase I (SPase) can change the specificity such that signal peptidase is able to cleave pro-OmpA nuclease A in vitro after phenylalanine or asparagine residues at the -1 position (Karla, A., Lively, M. O., Paetzel, M. and Dalbey, R. (2005) J. Biol. Chem. 280, 6731-6741). In this study, screening of a fluorescence resonance energy transfer-based peptide library revealed that the I144A, I144C, and I144C/I86T SPase mutants have a more relaxed substrate specificity at the -3 position, in comparison to the wild-type SPase. The double mutant tolerated arginine, glutamine, and tyrosine residues at the -3 position of the substrate. The altered specificity of the I144C/I86T mutant was confirmed by in vivo processing of pre-beta-lactamase containing non-canonical arginine and glutamine residues at the -3 position. This work establishes Ile-144 and Ile-86 as key P3 substrate specificity determinants for signal peptidase I and demonstrates the power of the fluorescence resonance energy transfer-based peptide library approach in defining the substrate specificity of proteases. PMID- 17077082 TI - Optical manipulation reveals strong attracting forces at membrane contact sites between endoplasmic reticulum and chloroplasts. AB - Eukaryote cells depend on membrane lipid trafficking from biogenic membranes, like the endoplasmic reticulum (ER), to other membranes in the cell. Two major routes for membrane lipid transport are recognized: vesicular trafficking and lipid transfer at zones of close contact between membranes. Specific ER regions involved in such membrane contact sites (MCSs) have been isolated, and lipid transfer at MCSs as well as protein-protein interactions between the partaking membranes have been demonstrated (reviewed by Holthuis, J. C. M., and Levine, T. P. (2005) Nat. Rev. 6, 209-220). Here we present the first demonstration of the physical association between membranes involved in MCSs: by using optical imaging and manipulation, strong attracting forces between ER and chloroplasts are revealed. We used Arabidopsis thaliana expressing green fluorescent protein in the ER lumen and observed leaf protoplasts by confocal microscopy. The ER network was evident, with ER branch end points apparently localized at chloroplast surfaces. After rupture of a protoplast using a laser scalpel, the cell content was released. ER fragments remained attached to the released chloroplasts and could be stretched out by optical tweezers. The applied force, 400 pN, could not drag a chloroplast free from its attached ER, which could reflect protein-protein interactions at the ER-chloroplast MCSs. As chloroplasts rely on import of ER synthesized lipids, we propose that lipid transfer occurs at these MCSs. We suggest that lipid transfer at the MCSs also occurs in the opposite direction, for example to channel plastid-synthesized acyl groups to supply substrates for ER-localized synthesis of membrane and storage lipids. PMID- 17077083 TI - Interaction of FoxO1 and TSC2 induces insulin resistance through activation of the mammalian target of rapamycin/p70 S6K pathway. AB - Both TSC2 (tuberin) and forkhead transcription factor FoxO1 are phosphorylated and inhibited by Akt and play important roles in insulin signaling. However, little is known about the relationship between TSC2 and FoxO1. Here we identified TSC2 as a FoxO1-binding protein by using a yeast two-hybrid screening with a murine islet cDNA library. Among FoxOs, only FoxO1 can be associated with TSC2. The physical association between the C terminus of TSC2 (amino acids 1280-1499) and FoxO1 degrades the TSC1-TSC2 complex and inhibits GTPase-activating protein activity of TSC2 toward Rheb. Overexpression of wild type FoxO1 enhances p70 S6K phosphorylation, whereas overexpression of TSC2 can reverse these effects. Knockdown of endogenous FOXO1 in human vascular endothelial cells decreased phosphorylation of p70 S6K. Prolonged overexpression of wild type FoxO1 enhanced phosphorylation of serine 307 of IRS1 and decreased phosphorylation of Akt and FoxO1 itself even in the presence of serum. These data suggest a novel mechanism by which FoxO1 regulates the insulin signaling pathway through negative regulation of TSC2 function. PMID- 17077084 TI - Structural and spectroscopic characterization of P450 BM3 mutants with unprecedented P450 heme iron ligand sets. New heme ligation states influence conformational equilibria in P450 BM3. AB - Two novel P450 heme iron ligand sets were generated by directed mutagenesis of the flavocytochrome P450 BM3 heme domain. The A264H and A264K variants produce Cys-Fe-His and Cys-Fe-Lys axial ligand sets, which were validated structurally and characterized by spectroscopic analysis. EPR and magnetic circular dichroism (MCD) provided fingerprints defining these P450 ligand sets. Near IR MCD spectra identified ferric low spin charge-transfer bands diagnostic of the novel ligands. For the A264K mutant, this is the first report of a Cys-Fe-Lys near-IR MCD band. Crystal structure determination showed that substrate-free A264H and A264K proteins crystallize in distinct conformations, as observed previously in substrate-free and fatty acid-bound wild-type P450 forms, respectively. This, in turn, likely reflects the positioning of the I alpha helix section of the protein that is required for optimal configuration of the ligands to the heme iron. One of the monomers in the asymmetric unit of the A264H crystals was in a novel conformation with a more open substrate access route to the active site. The same species was isolated for the wildtype heme domain and represents a novel conformational state of BM3 (termed SF2). The "locking" of these distinct conformations is evident from the fact that the endogenous ligands cannot be displaced by substrate or exogenous ligands. The consequent reduction of heme domain conformational heterogeneity will be important in attempts to determine atomic structure of the full-length, multidomain flavocytochrome, and thus to understand in atomic detail interactions between its heme and reductase domains. PMID- 17077085 TI - Self-chaperoning of the type III secretion system needle tip proteins IpaD and BipD. AB - Bacteria expressing type III secretion systems (T3SS) have been responsible for the deaths of millions worldwide, acting as key virulence elements in diseases ranging from plague to typhoid fever. The T3SS is composed of a basal body, which traverses both bacterial membranes, and an external needle through which effector proteins are secreted. We report multiple crystal structures of two proteins that sit at the tip of the needle and are essential for virulence: IpaD from Shigella flexneri and BipD from Burkholderia pseudomallei. The structures reveal that the N-terminal domains of the molecules are intramolecular chaperones that prevent premature oligomerization, as well as sharing structural homology with proteins involved in eukaryotic actin rearrangement. Crystal packing has allowed us to construct a model for the tip complex that is supported by mutations designed using the structure. PMID- 17077086 TI - Cysteine-scanning analysis of the nucleobase-ascorbate transporter signature motif in YgfO permease of Escherichia coli: Gln-324 and Asn-325 are essential, and Ile-329-Val-339 form an alpha-helix. AB - The nucleobase-ascorbate transporter (NAT) signature motif is a conserved sequence motif of the ubiquitous NAT/NCS2 family implicated in defining the function and selectivity of purine translocation pathway in the major fungal homolog UapA. To analyze the role of NAT motif more systematically, we employed Cys-scanning mutagenesis of the Escherichia coli xanthine-specific homolog YgfO. Using a functional mutant devoid of Cys residues (C-less), each amino acid residue in sequence (315)GSIPITTFAQNNGVIQMTGVASRYVG(340) (motif underlined) was replaced individually with Cys. Of the 26 single-Cys mutants, 16 accumulate xanthine to > or =50% of the steady state observed with C-less YgfO, 4 accumulate to low levels (10-25% of C-less), F322C, N325C, and N326C accumulate marginally (5-8% of C-less), and P318C, Q324C, and G340C are inactive. When transferred to wild type, F322C(wt) and N326C(wt) are highly active, but P318G(wt), Q324C(wt), N325C(wt), and G340C(wt) are inactive, and G340A(wt) displays low activity. Immunoblot analysis shows that replacements at Pro-318 or Gly-340 are associated with low or negligible expression in the membrane. More extensive mutagenesis reveals that Gln-324 is critical for high affinity uptake and ligand recognition, and Asn-325 is irreplaceable for active xanthine transport, whereas Thr-332 and Gly-333 are important determinants of ligand specificity. All single-Cys mutants react with N-ethylmaleimide, but regarding sensitivity to inactivation, they fall to three regions; positions 315-322 are insensitive to N-ethylmaleimide, with IC(50) values > or =0.4 mM, positions 323-329 are highly sensitive, with IC(50) values of 15-80 microM, and sensitivity of positions 330-340 follows a periodicity, with mutants sensitive to inactivation clustering on one face of an alpha-helix. PMID- 17077087 TI - p53 suppresses the Nrf2-dependent transcription of antioxidant response genes. AB - Cells respond to the shift of intracellular environment toward pro-oxidant conditions by activating the transcription of numerous "antioxidant" genes. This response is based on the activation of the Nrf2 transcription factor, which transactivates the genes containing in their promoters the antioxidant response cis-elements (AREs). If the oxidative stress provokes DNA damage, a second response of the cell takes place, based on the activation of p53, which induces cell cycle arrest and/or apoptosis. Here we have explored the cross-talk between these two regulatory mechanisms. The results show that p53 counteracts the Nrf2 induced transcription of three ARE-containing promoters of the x-CT, NQO1, and GST-alpha1 genes. Endogenous transcripts of these antioxidant genes accumulate as a consequence of Nrf2 overexpression or exposure to electrophile diethylmaleate, but these effects are again blocked by p53 overexpression or endogenous p53 activation. Chromatin immunoprecipitation experiments support the hypothesis that this p53-dependent trans-repression is due to the direct interaction of p53 with the ARE-containing promoters. Considering that p53-induced apoptosis requires an accumulation of reactive oxygen species, this negative control on the Nrf2 transactivation appears to be aimed to prevent the generation of a strong anti oxidant intracellular environment that could hinder the induction of apoptosis. PMID- 17077088 TI - Structural and functional properties of two human FXYD3 (Mat-8) isoforms. AB - Six of 7 FXYD proteins have been shown to be tissue-specific modulators of Na,K ATPase. In this study, we have identified two splice variants of human FXYD3, or Mat-8, in CaCo-2 cells. Short human FXYD3 has 72% sequence identity with mouse FXYD3, whereas long human FXYD3 is identical to short human FXYD3 but has a 26 amino acid insertion after the transmembrane domain. Short and long human FXYD3 RNAs and proteins are differentially expressed during differentiation of CaCo-2 cells. Long human FXYD3 is mainly expressed in nondifferentiated cells and short human FXYD3 in differentiated cells and both FXYD3 variants can be co immunoprecipitated with a Na,K-ATPase antibody. In contrast to mouse FXYD3, which has two transmembrane domains for lack of cleavage of the signal peptide, human FXYD3 has a cleavable signal peptide and adopts a type I topology. After co expression in Xenopus oocytes, both human FXYD3 variants associate stably only with Na,K-ATPase isozymes but not with H,K-ATPase or Ca-ATPase. Similar to mouse FXYD3, short human FXYD3 decreases the apparent K(+) and Na(+) affinity of Na,K ATPase over a large range of membrane potentials. On the other hand, long human FXYD3 decreases the apparent K(+) affinity only at slightly negative and positive membrane potentials and increases the apparent Na(+) affinity of Na,K-ATPase. Finally, both short and long human FXYD3 induce a hyperpolarization activated current, similar to that induced by mouse FXYD3. Thus, we have characterized two human FXYD3 isoforms that are differentially expressed in differentiated and non differentiated cells and show different functional properties. PMID- 17077089 TI - Arg-158 is critical in both binding the substrate and stabilizing the transition state oxyanion for the enzymatic reaction of malonamidase E2. AB - Malonamidase E2 (MAE2) from Bradyrhizobium japonicum is an enzyme that hydrolyzes malonamate to malonate and has a Ser-cis-Ser-Lys catalytic triad at the active site. The crystal structures of wild type and mutant MAE2 exhibited that the guanido group of Arg-158 could be involved in the binding of malonamate in which the negative charge of the carboxyl group could destabilize a negatively charged transition-state oxyanion in the enzymatic reaction. In an attempt to elucidate the specific roles of Arg-158, site-directed mutants, R158Q, R158E, and R158K, were prepared (see Table 1). The crystal structure of R158Q determined at 2.2 Angstrom resolution showed that the guanido group of Arg-158 was important for the substrate binding with the marginal structural change upon the mutation. The k(cat) value of R158Q significantly decreased by over 1500-fold and the catalytic activity of R158E could not be detected. The k(cat) value of R158K was similar to that of the wild type with the K(m) value drastically increased by 100-fold, suggesting that Lys-158 of R158K can stabilize the negative charge of the carboxylate in the substrate to some extent and contribute to the stabilization of the transition-state oxyanion, but a single amine group of Lys-158 in R158K could not precisely anchor the carboxyl group of malonamate compared with the guanido group of Arg-158. Our kinetic and structural evidences demonstrate that Arg-158 in MAE2 should be critical to both binding the substrate and stabilizing the transition-state oxyanion for the catalytic reaction of MAE2. PMID- 17077090 TI - The Drosophila heterogeneous nuclear ribonucleoprotein M protein, HRP59, regulates alternative splicing and controls the production of its own mRNA. AB - The Drosophila heterogeneous nuclear ribonucleoprotein M, HRP59, is a nuclear protein that associates co-transcriptionally with pre-mRNA and is necessary for the correct expression of a subset of mRNAs. We show here that the hrp59 pre-mRNA is alternatively spliced to generate two different mRNAs that differ in the presence of exon 3. Exon 3-containing transcripts make up the majority of hrp59 transcripts and encode for the functional protein, HRP59-1. Transcripts that lack exon 3 contain a premature translation termination codon and are targeted to the nonsense mediated decay pathway. We show that exon 3 inclusion is itself inhibited by HRP59 and that changes in the HRP59 protein levels affect the splicing activity of the cell. We propose that the ability of HRP59 to regulate the alternative splicing of its own pre-mRNA serves in a negative feedback loop that controls the levels of the HRP59 protein and maintains the homeostasis of the splicing environment. PMID- 17077091 TI - Influence of environmental changes on integrating radon detectors: results of an intercomparison exercise. AB - An intercomparison exercise for passive integrating radon detectors has been carried out with the participation of 12 detection systems from 10 laboratories. The detection systems comprise three commonly used in radon integrating measurements, tracks, activated charcoal canisters and electrets. The exposures were carried out in the radon and thoron chambers at the Institute of Energy Techniques (INTE) of the Technical University of Catalonia (UPC), which is considered to be the Spanish reference chamber. The detectors were exposed to three different temperatures (10, 20 and 30 degrees C) and relative humidities (30, 45 and 80%). Furthermore, in three exposures radon concentration was drastically changed during the exposure period in order to study the efficiency of canister collection. The results indicated that only the charcoal canister response was found to be significantly influenced by external climatic conditions and radon fluctuations. Those track detectors, which are unable to measure thoron concentrations show thoron sensitivity and thus interfere with precise measurement of radon. Detectors for measuring thoron concentration show quite a different response, which could be related to their traceability. PMID- 17077092 TI - Scanning an individual monitoring database for multiple occurrences using bi-gram analysis. AB - Maintaining the integrity of the databases is one of the important aspects of quality assurance at individual monitoring services and national dose registers. This paper presents a method for finding and preventing the occurrence of duplicate entries in the databases that can occur, e.g. because of a variable spelling or misspelling of the name. The method is based on bi-gram text analysis techniques. The methods can also be used for retrieving dose data in historical databases in the framework of dose reconstruction efforts of persons of whom the spelling of the name as originally entered, possibly decades ago, is uncertain. PMID- 17077093 TI - Evaluation of the photoneutron field produced in a medical linear accelerator. AB - The doses and spectra of photoneutrons produced in a medical linear accelerator with photon energies of 10 and 15 MV were evaluated. The Monte Carlo code, MCNPX, was used to simulate the transport of these photoneutrons around the head for 10 and 15 MV photons. The fully-described geometry of the accelerator head was used in this calculation. The photoneutron energy spectra and doses for various photon field sizes were calculated at each of 20 positions. The results indicate that the maximum dose equivalents are observed in 20 x 20 cm(2) case among photon fields. It was found the neutron average energy at isocenter for a 0 x 0 cm(2) field is 0.38 MeV for 10 MV and is 0.45 MeV for 15 MV. The neutron doses at 10 positions around the head in the treatment room of the operation facility at 10 and 15 MV were measured using the bubble detectors. Measurements were compared with the calculations under the same geometry in the experiment. It was found that the majority of the calculated results agreed to within the standard deviations of the measurements. These above results can be applied in the verification of maximum allowed neutron leakage percentage of treatment dose defined in the IEC. We have been employing them to derive the empirical formula for neutron dose equivalent level at the maze entrance of medical accelerator treatment rooms in a study that is still underway. PMID- 17077094 TI - Indoor radon in rural dwellings of the South-Pannonian region. AB - The results of indoor radon survey in the South-Pannonian Province Vojvodina (Serbia and Montenegro) are presented. The sampling strategy was oriented towards suburban and urban regions in the Province. For the dwellings typical for such regions the geometric mean annual radon activity concentration of 76.1 Bq m(-3) is measured (1000 measurements). This result leads to the annual dose estimate of 4.3 mSv y(-1), which is above the recommended action limit of ICRP. For urban dwellings in Novi Sad (the Province capital), the annual mean value of 54 Bq m( 3) (220 measurements) is obtained. By comparison of these two results it is concluded that radon surveys based on measurements in urban environment may seriously underestimate the radon-related health risk. The elevated radon levels could not be explained by elevated uranium levels of surface soil. PMID- 17077095 TI - Dependence network modeling for biomarker identification. AB - MOTIVATION: Our purpose is to develop a statistical modeling approach for cancer biomarker discovery and provide new insights into early cancer detection. We propose the concept of dependence network, apply it for identifying cancer biomarkers, and study the difference between the protein or gene samples from cancer and non-cancer subjects based on mass-spectrometry (MS) and microarray data. RESULTS: Three MS and two gene microarray datasets are studied. Clear differences are observed in the dependence networks for cancer and non-cancer samples. Protein/gene features are examined three at one time through an exhaustive search. Dependence networks are constructed by binding triples identified by the eigenvalue pattern of the dependence model, and are further compared to identify cancer biomarkers. Such dependence-network-based biomarkers show much greater consistency under 10-fold cross-validation than the classification-performance-based biomarkers. Furthermore, the biological relevance of the dependence-network-based biomarkers using microarray data is discussed. The proposed scheme is shown promising for cancer diagnosis and prediction. AVAILABILITY: See supplements: http://dsplab.eng.umd.edu/~genomics/dependencenetwork/ PMID- 17077096 TI - Simple sequence repeats in organellar genomes of rice: frequency and distribution in genic and intergenic regions. AB - MOTIVATION: Simple sequence repeats (SSRs) are abundant across genomes. However, the significance of SSRs in organellar genomes of rice has not been completely understood. The availability of organellar genome sequences allows us to understand the organization of SSRs in their genic and intergenic regions. RESULTS: We have analyzed SSRs in mitochondrial and chloroplast genomes of rice. We identified 2528 SSRs in the mitochondrial genome and average 870 SSRs in the chloroplast genomes. About 8.7% of the mitochondrial and 27.5% of the chloroplast SSRs were observed in the genic region. Dinucleotides were the most abundant repeats in genic and intergenic regions of the mitochondrial genome while mononucleotides were predominant in the chloroplast genomes. The rps and nad gene clusters of mitochondria had the maximum repeats, while the rpo and ndh gene clusters of chloroplast had the maximum repeats. We identified SSRs in both organellar genomes and validated in different cultivars and species. PMID- 17077097 TI - OmicBrowse: a browser of multidimensional omics annotations. AB - OmicBrowse is a browser to explore multiple datasets coordinated in the multidimensional omic space integrating omics knowledge ranging from genomes to phenomes and connecting evolutional correspondences among multiple species. OmicBrowse integrates multiple data servers into a single omic space through secure peer-to-peer server communications, so that a user can easily obtain an integrated view of distributed data servers, e.g. an integrated view of numerous whole-genome tiling-array data retrieved from a user's in-house private-data server, along with various genomic annotations from public internet servers. OmicBrowse is especially appropriate for positional-cloning purposes. It displays both genetic maps and genomic annotations within wide chromosomal intervals and assists a user to select candidate genes by filtering their annotations or associated documents against user-specified keywords or ontology terms. We also show that an omic-space chart effectively represents schemes for integrating multiple datasets of multiple species. AVAILABILITY: OmicBrowse is developed by the Genome-Phenome Superbrain Project and is released as free open-source software under the GNU General Public License at http://omicspace.riken.jp. PMID- 17077098 TI - GECO--linear visualization for comparative genomics. AB - In order to understand and interpret phylogenetic and functional relationships between multiple prokaryotic species, qualitative and quantitative data must be correlated and displayed. GECO allows linear visualization of multiple genomes using a client/server based approach by dynamically creating .png- or .pdf formatted images. It is able to display ortholog relations calculated using BLASTCLUST by color coding ortholog representations. Irregularities on the genomic level can be identified by anomalous G/C composition. Thus, this software will enable researchers to detect horizontally transferred genes, pseudogenes and insertions/deletions in related microbial genomes. AVAILABILITY: http://bioinfo.mikrobio.med.uni-giessen.de/geco2/GecoMainServlet PMID- 17077099 TI - Towards clustering of incomplete microarray data without the use of imputation. AB - MOTIVATION: Clustering technique is used to find groups of genes that show similar expression patterns under multiple experimental conditions. Nonetheless, the results obtained by cluster analysis are influenced by the existence of missing values that commonly arise in microarray experiments. Because a clustering method requires a complete data matrix as an input, previous studies have estimated the missing values using an imputation method in the preprocessing step of clustering. However, a common limitation of these conventional approaches is that once the estimates of missing values are fixed in the preprocessing step, they are not changed during subsequent processes of clustering; badly estimated missing values obtained in data preprocessing are likely to deteriorate the quality and reliability of clustering results. Thus, a new clustering method is required for improving missing values during iterative clustering process. RESULTS: We present a method for Clustering Incomplete data using Alternating Optimization (CIAO) in which a prior imputation method is not required. To reduce the influence of imputation in preprocessing, we take an alternative optimization approach to find better estimates during iterative clustering process. This method improves the estimates of missing values by exploiting the cluster information such as cluster centroids and all available non-missing values in each iteration. To test the performance of the CIAO, we applied the CIAO and conventional imputation-based clustering methods, e.g. k-means based on KNNimpute, for clustering two yeast incomplete data sets, and compared the clustering result of each method using the Saccharomyces Genome Database annotations. The clustering results of the CIAO method are more significantly relevant to the biological gene annotations than those of other methods, indicating its effectiveness and potential for clustering incomplete gene expression data. AVAILABILITY: The software was developed using Java language, and can be executed on the platforms that JVM (Java Virtual Machine) is running. It is available from the authors upon request. PMID- 17077100 TI - Estimating p-values in small microarray experiments. AB - MOTIVATION: Microarray data typically have small numbers of observations per gene, which can result in low power for statistical tests. Test statistics that borrow information from data across all of the genes can improve power, but these statistics have non-standard distributions, and their significance must be assessed using permutation analysis. When sample sizes are small, the number of distinct permutations can be severely limited, and pooling the permutation derived test statistics across all genes has been proposed. However, the null distribution of the test statistics under permutation is not the same for equally and differentially expressed genes. This can have a negative impact on both p value estimation and the power of information borrowing statistics. RESULTS: We investigate permutation based methods for estimating p-values. One of methods that uses pooling from a selected subset of the data are shown to have the correct type I error rate and to provide accurate estimates of the false discovery rate (FDR). We provide guidelines to select an appropriate subset. We also demonstrate that information borrowing statistics have substantially increased power compared to the t-test in small experiments. PMID- 17077101 TI - Effect of quinolone prophylaxis in afebrile neutropenic patients on microbial resistance: systematic review and meta-analysis. AB - OBJECTIVES: To assess the effect of quinolone prophylaxis following chemotherapy for malignancies on the emergence of resistant bacteria in neutropenic patients. METHODS: Systematic review and meta-analysis of randomized controlled trials comparing quinolone prophylaxis with placebo or no intervention, or another antibiotic, for the prevention of bacterial infections in afebrile neutropenic patients. The Cochrane Library, PubMed, Embase, conference proceedings and references were searched. Two reviewers independently applied selection criteria, carried out quality assessment and extracted the data. Relative risks (RR) with 95% confidence intervals (CIs) were estimated and pooled. Primary outcomes were rates of colonization and infection by quinolone-resistant bacteria. RESULTS: The search yielded 56 trials, 22 compared quinolones with placebo or no intervention. Data on colonization by resistant organisms could be extracted from 27 trials (48%). When compared with placebo or no intervention, there was a statistically non-significant increase in colonization with organisms resistant to quinolones (RR 1.68; 95% CI 0.71-4.00). There was no difference in the number of patients developing infections caused by resistant pathogens (RR 1.04; 95% CI 0.73-1.50). In trials comparing quinolones with trimethoprim/sulfamethoxazole, there were fewer incidents of colonization by bacteria resistant to the prophylactic agent in the quinolone arm than in the trimethoprim/sulfamethoxazole arm (RR 0.49; 95% CI 0.37-0.66). Data on baseline resistance of colonizing isolates, resistance development and cross-resistance to beta-lactam antibiotics were too scarce to analyse. CONCLUSIONS: Patients treated with quinolones have a non-significant increase in colonization by quinolone-resistant bacteria. There is no difference in the number of infections caused by pathogens resistant to quinolones. As quinolone prophylaxis reduces the risk of death in neutropenic patients, the risk associated with colonization and infections caused by quinolone-resistant organisms does not outweigh the gain. Future trials should focus on better documentation of infections caused by resistant organisms. PMID- 17077102 TI - Association between availability of heroin and methadone and fatal poisoning in England and Wales 1993-2004. AB - BACKGROUND: The UK heroin market is the biggest in Europe and approximately 70% of heroin deaths are due to fatal poisoning. Methadone treatment for heroin addiction in the UK, the 'British system', is unique as it is largely provided by General Practitioners. METHODS: The Office for National Statistics provided data on deaths, the Home Office provided law enforcement data on drug seizures and the Department of Health data on prescriptions. For methadone treatment we calculated the death rate per 1000 patient years. We used Spearman's rank correlation to assess the association between illicit drug seizures for heroin and methadone and deaths. RESULTS: Between 1993 and 2004 there were 7072 deaths involving heroin/morphine (86% males) and 3298 deaths involving methadone (83% male). From 1993-1997, directly age-standardized mortality rates for males were similar for both drugs, increasing from approximately 5 to 15 per million. Mortality rates for heroin continued to increase until 2000, subsequently decreasing from 30 to 20 per million by 2003, and rising again to 24 per million in 2004. In contrast, mortality rates for methadone decreased between 1997 and 2004 to just above 1993 levels. Among females the mortality rate for both drugs was lower than for males throughout the study period, remaining relatively stable. Methadone deaths per 1000 patient years remained similar between 1993 and 1997, after which they fell by three quarters. For both heroin/morphine and methadone, deaths were strongly associated with seizures (Spearmans' coefficient for males: heroin, P = 0.95, P < 0.001 and methadone, P = 0.83, P = 0.0013). CONCLUSIONS: Our study suggests the 'British System' can deliver substantial expansion of treatment without increased mortality risk. The fall in heroin/morphine deaths since 2000 may also be an indication of success of increasing methadone treatment. Data on mortality risk is needed to determine whether increased methadone treatment has reduced drug related deaths. PMID- 17077103 TI - Commentary: proportion not the cause of beauty but of health? PMID- 17077105 TI - Subthalamic stimulation modulates cortical control of urinary bladder in Parkinson's disease. AB - Subthalamic nucleus deep brain stimulation (STN-DBS) is an effective therapy for off-period motor symptoms and dyskinesias in advanced Parkinson's disease. Clinical studies have shown that STN-DBS also ameliorates urinary bladder function in Parkinson's disease patients by delaying the first desire to void and increasing bladder capacity. This study aimed at investigating the effect of STN DBS on the neural mechanisms underlying cerebral bladder control. Using PET to measure changes in regional cerebral blood flow (rCBF), 11 patients with bilateral STN-DBS were studied during urodynamic bladder filling in STN-DBS ON and OFF condition. A filled bladder led to a significant increase of rCBF in the anterior cingulate cortex, which was further enhanced during STN-DBS OFF. A significant interaction between bladder state and STN-DBS was observed in lateral frontal cortex with increased rCBF when the bladder was filled during STN-DBS OFF. The data suggest that STN-DBS ameliorates bladder dysfunction and that this modulation may result from facilitated processing of afferent bladder information. PMID- 17077106 TI - Occupational exposure during application and removal of antifouling paints. AB - Exposure data on biocides are relatively rare in published literature, especially for secondary exposure. This is also the case for antifouling exposure. Therefore, a field study was carried out measuring exposure to antifouling paints. Both primary exposure (rolling and spraying) and secondary exposure (during sand blasting) were studied. Exposure during rolling was measured in boatyards where paints containing dichlofluanid (DCF) were applied. Spraying was measured in dockyards (larger than boatyards) where paints containing copper were applied. Furthermore, during sand blasting the removal of old paint layers containing copper was measured. A total of 54 datasets was collected, both for inhalation and dermal exposure data. For paint and stripped paint bulk analyses were performed. The following values are all arithmetic means of the datasets. Inhalation of copper amounted to 3 mg m-3 during spraying and to 0.8 mg m-3 during sand blasting. Potential body exposure loading amounted to 272 mg h-1 copper during spraying and 33 mg h-1 during sand blasting. For dichlofluanid the inhalation exposure loading was 0.14 mg m-3 during rolling, whereas the potential body exposure loading was 267 mg h-1 and potential hand exposure loading 277 mg h 1. The results for primary exposure compare well to the very few public data available. For the secondary exposure (sand blasting) no comparable data were available. The present study shows that the exposure loading should be considered more extensively, including applicable protective gear. In this light the findings for the potmen during sand blasting suggest that personal protective equipment should be (re)considered carefully. PMID- 17077107 TI - Increased telomerase activity and human telomerase reverse transcriptase mRNA expression in the endometrium of patients with endometriosis. AB - BACKGROUND: Endometriosis is considered a frequent, benign disease with the ability to undergo neoplastic processes. The aim of this study was to evaluate the limitless replication potential of the endometrium in patients with endometriosis by examining human telomerase reverse transcriptase (hTERT) mRNA expression and telomerase activity. METHODS: Endometrium samples from 30 endometriosis patients and 30 patients without endometriosis were obtained via endometrial biopsy. The expression of hTERT mRNA was determined by real-time RT PCR assay, and telomerase activity was measured by telomerase repeat amplification protocol (TRAP) assay. RESULTS: The mean normalized hTERT (N hTERT) mRNA level was significantly higher in the endometriosis than in the control group (P = 0.013). The mean hTERT mRNA levels during the proliferative phase and during the secretory phase were higher in the endometriosis group than in the control group, although the difference was only significant for the secretory phase (P = 0.036). We found a prominent difference in endometrial telomerase activity between moderate-to-severe endometriosis and the control group (P = 0.048). The levels of hTERT mRNA and telomerase activity increased as the disease became more severe (P = 0.038, P = 0.016). CONCLUSIONS: This study showed the overexpression of hTERT mRNA and telomerase activity in the endometrium of endometriosis patients. These finding suggest that replication potential of endometrial cells may have an important role in the pathogenesis of endometriosis. PMID- 17077108 TI - Development of a device for transcatheter pulmonary artery banding: evaluation in animals. AB - AIMS: Pulmonary artery banding (PAB) is the first palliation in infants with complex congenital heart disease and elevated pulmonary blood flow. In older patients with corrected transposition of the great arteries, it may be used to re train the left ventricle. To date, the only option is surgical. We report the development and the evaluation of a device for transcatheter PAB. METHODS AND RESULTS: We intended to implant a pulmonary artery (PA) reducer percutaneously between the native pulmonic valve and the pulmonary bifurcation. Immediately following its insertion, we planned to implant a balloon expandable stent inside the restriction to calibrate the banding. Sheep were sacrificed acutely (group 1, n=6) and after 1 month of follow-up (group 2, n=6), the reducer was implanted successfully in all animals. It allowed the PA diameter to be reduced from 25 to 10.5 mm. Bare stents were successfully delivered inside the reducer. No paraprosthetic leak was found by injecting contrast dye. After the insertion procedure, signs of intolerance to obstruction were present in all animals and prompted us to dilate the stents from 12 to 16 mm. One animal from group 1 died before a balloon dilatation could be achieved. In the animals from group 2, the mean systolic gradient was 19 and 34.8 mmHg, respectively, at early and late evaluation. CONCLUSION: Implantation of a PA reducer is possible in sheep, through a transcatheter approach allowing intravascular PAB. Miniaturization of the device is necessary to enlarge its use from adulthood to childhood. PMID- 17077109 TI - Multiple myocardial abscesses successfully treated with medical management in an immunosuppressed patient. PMID- 17077110 TI - Sperm-FISH analysis in a pericentric chromosome 1 inversion, 46,XY,inv(1)(p22q42), associated with infertility. AB - No phenotypic effect is observed in most inversion heterozygotes. However, reproductive risks may occur in the form of infertility, spontaneous abortions or chromosomally unbalanced children as a consequence of meiotic recombination between inverted and non-inverted chromosomes. An odd number of crossovers within the inverted segment results in gametes bearing recombinant chromosomes with a duplication of the region outside of the inversion segment of one arm and a deletion of the terminal segment of the other arm [dup(p)/del(q) and del(p)/dup(q)]. Using fluorescence in-situ hybridization (FISH), the chromosome segregation of a pericentric inversion of chromosome 1 was studied in spermatozoa of a inv(1)(p22q42) heterozygous carrier. Three-colour FISH was performed on sperm samples using a probe mixture consisting of chromosome 1p telomere-specific probe, chromosome 1q telomere-specific probe and chromosome 18 centromere specific alpha satellite DNA probe. The frequency of the non-recombinant product was 80.1%. The frequencies of the two types of recombinants carrying a duplication of the short arm and a deletion of the long arm, and vice versa, were respectively 7.6 and 7.2%, and these frequencies were not statistically significant from the expected ratio of 1:1. Sperm-FISH allows the further understanding of segregation patterns and their effect on reproductive failure and allows an accurate genetic counselling. PMID- 17077111 TI - Another public health triumph. PMID- 17077112 TI - Methamphetamine detection in maternal and neonatal hair: implications for fetal safety. AB - BACKGROUND: Methamphetamine misuse is a serious health problem of epidemic proportions. Use of this drug, particularly during pregnancy, is difficult to ascertain. Sparse information is available on gestational exposure. OBJECTIVES: To quantify methamphetamine accumulation in hair, identify the use of methamphetamine with other drugs of abuse and characterise correlations between concentrations of methamphetamine in maternal and neonatal hair. SUBJECTS AND METHODS: Motherisk laboratory at the Hospital for Sick Children routinely carries out analysis of methamphetamine in hair. Mothers and infants with positive results for methamphetamine in hair were identified. Drugs present in hair were analysed by ELISA and positive results were confirmed by gas chromatgraphy/mass spectrometry. RESULTS: 396 people positive for methamphetamine in their hair were identified from our database. Almost 85% of them were positive for at least one other drug of abuse, mostly cocaine. Eleven mother-baby pairs with hair positive for methamphetamine were identified. Methamphetamine levels in hair ranged between 0.13 and 51.97 ng/mg in the mothers and between 0 and 22.73 ng/mg in the neonates. Methamphetamine levels in mothers and neonates correlated significantly. One (9%) neonate was negative for methamphetamine even though the mother was positive. CONCLUSION: To our knowledge, this is the first report on fetal exposure to methamphetamine during pregnancy, showing transplacental transfer of the drug, with accumulation in fetal hair. Hair measurement for methamphetamine in neonates is a useful screening method to detect intra-uterine exposure to the drug. The data also indicate that positive exposure to methamphetamine strongly suggests that the person is a polydrug user, which may have important implications for fetal safety. PMID- 17077113 TI - An evaluation of optometrists' ability to correctly identify and manage patients with ocular disease in the accident and emergency department of an eye hospital. AB - AIM: To assess optometrists' ability to correctly identify and manage patients with different ocular conditions seen in the accident and emergency (A&E) department of an eye hospital. METHODS: Randomly selected patients presenting to the A&E department were initially examined by one of two senior optometrists and a consultant ophthalmologist. A diagnosis and a management plan were made for each patient by the optometrist and by the consultant, who was blinded to the optometrists' plan. Agreement was assessed between optometrist and consultant ophthalmologist for primary and secondary diagnoses, management plan and whether patients could be seen by an optometrist only. Weighted kappa (kappa) statistics was used to assess the level of agreement in management between the two groups. RESULTS: A total of 150 patients were assessed. The agreement in primary diagnosis and management outcome between the two groups were 89.3% (95% CI 83.2% to 93.8%) and 79.3% (95% CI 84.8% to 94.8%), respectively. A high level of agreement in management outcome was found (kappa = 0.82). No sight-threatening conditions were misdiagnosed by the optometrist. CONCLUSIONS: There was good agreement in both the diagnosis and management plan between optometrists and the ophthalmologist. This study has shown that optometrists can potentially work safely in an A&E department of a busy eye hospital. PMID- 17077114 TI - Pretreatment of posterior subtenon injection of triamcinolone acetonide has beneficial effects for grid pattern photocoagulation against diffuse diabetic macular oedema. AB - AIM: To prospectively evaluate the efficacy of subtenon injection of triamcinolone acetonide (TA) before laser grid pattern photocoagulation (G-PC) for the treatment of diffuse diabetic macular oedema (DDME). METHODS: 42 eyes of 37 consecutive patients with DDME were studied. 1 week before G-PC, 21 eyes received TA subtenon injection, and the other eyes served as control. The clinical course of visual acuity (VA) and foveal thickness (FT) was monitored for up to 24 weeks after G-PC. Mean deviation (MD) of perimetry with 30-2 program on Humphrey Perimeter (Zeiss-Humphrey, Dublin, California, USA) was also measured. The average laser intensity was recorded. RESULTS: After TA injection, FT and VA were improved, and subsequent G-PC maintained the improvement for up to 24 weeks without recurrence of diffuse diabetic macular oedema. In contrast, G-PC without TA injection induced transient worsening of FT and VA, then both were gradually improved. At 24 weeks after G-PC, MD in the TA-injected eyes was better than those in control. The required laser intensity in TA-injected eyes was less than that for control. CONCLUSION: Subtenon injection of TA prior to G-PC allows for treatment with a lower intensity of laser spots and also prevents the decrease in central visual field sensitivity, all of which have clinical advantages for G-PC. PMID- 17077115 TI - The prevalence and severity of diabetic retinopathy, associated risk factors and vision loss in patients registered with type 2 diabetes in Luganville, Vanuatu. AB - AIM: To determine the prevalence and severity of diabetic retinopathy in patients with type 2 diabetes in Luganville, the second largest town in Vanuatu. Additionally, to investigate risk factors for retinopathy and the effect of retinopathy on visual acuity (VA) within this group. METHOD: All 83 registered patients with type 2 diabetes in Luganville, a town of 13 121 people, were invited for an interview and anthropometric measurements. A questionnaire including assessment of hypertension and glycaemic control, which are known risk factors for diabetic retinopathy, was administered. This sample accounted for approximately 1.07% of Luganville's adult population. Presenting VA was measured. The retina was photographed with a non-mydriatic fundus camera and images later independently graded for the extent of retinopathy. RESULTS: 68 (82%) of the 83 patients attended. The mean (SD) age was 54 (11) years and 31 (46%) were male. Diabetic retinopathy was present in 36 (52.9%) of the sample. Sight-threatening retinopathy requiring urgent referral was present in 15 (22.1%) patients. Presenting VA was worse than 6/12 in the better eye in n = 32 (47%) and in up to half of these cases the principal cause was retinopathy. In addition, four people had uniocular blindness resulting from diabetes. The mean body mass index was lower in those patients with diabetes with retinopathy than in those without (p = 0.010), but there were no other significant differences between the two groups and, specifically, no difference in the frequency of retinopathy risk factors. 42 (61.8%) patients had hypertension (>or=135/85 mm Hg) or were taking antihypertensive therapy. CONCLUSIONS: The prevalence of registered patients with diabetes in Luganville's adult population was 1.07%. Diabetic retinopathy was highly prevalent in the sample (in 36, 52.9%), and in 15 (22.1%) there was a significant threat to sight, with up to 25% of the sample possibly already affected by decreased VA or blindness resulting from diabetes-related eye disease. Retinopathy risk factors were also prevalent. A diabetes screening programme with baseline ophthalmic assessment and follow-up are urgently needed to enable timely intervention and treatment. PMID- 17077116 TI - Physical activity and the 15-year cumulative incidence of age-related macular degeneration: the Beaver Dam Eye Study. AB - BACKGROUND: Cardiovascular disease and age-related macular degeneration (AMD) may share common risk factors. Physical activity improves the cardiovascular risk profile; however, there have been few studies investigating a relationship between physical activity and the long-term incidence of AMD. METHODS: The 15 year cumulative incidence of AMD was determined through four examination phases at 5-year intervals of a population-based study conducted in Beaver Dam, Wisconsin, USA, initiated in 1988-90 (n = 3874 men and women between ages 43 and 86 years). Early AMD (pigment abnormalities or soft indistinct drusen), exudative AMD and geographic atrophy were determined by grading stereoscopic colour fundus photographs. Measures of physical activity were obtained through a questionnaire administered at the baseline examination. RESULTS: After controlling for age, sex, history of arthritis, systolic blood pressure, body mass index, smoking and education, people with an active lifestyle (defined as regular activity > or =3 times/week) at baseline were less likely to develop exudative AMD (odds ratio (OR) 0.3, 95% confidence interval (CI) 0.1 to 0.7) compared with people without an active lifestyle. After multivariate adjustment, increased categories of number of blocks walked per day decreased the risk of exudative AMD (OR 0.7, 95% CI 0.6 to 0.97). Physical activity was not related to the incidence of early AMD or pure geographic atrophy. CONCLUSIONS: These data show a protective effect of physical activity for incident exudative AMD, independent of body mass index and other confounders. They also suggest a possible modifiable behaviour that might be protective against developing AMD. PMID- 17077117 TI - Visual quality of life after macular hole surgery: outcome and predictive factors. AB - BACKGROUND: In the present study we evaluated the functional success after macular hole surgery in correlation to visual quality of life and looked for predictive factors determining surgical success. METHODS: Fifty-nine patients that underwent pars plana vitrectomy for idiopathic macular hole were included. Follow-up visits were performed in regular intervals after surgery and included a clinical examination, optical coherence tomography (OCT) and measurement of visual acuity. To assess the visual quality of life patients filled out the National Eye Institute 25-item Visual Function Questionnaire (VFQ-25) before and three months and one year after surgery. RESULTS: Macular hole closure was achieved in 57 of 59 patients (97%). Mean visual acuity increased from 20/100 preoperatively to 20/34 one year after surgery (p = 0.02). Despite good visual acuity (20/27) in the fellow eye, visual quality of life (VFQ composite score) rose from 75.9 +/- 14.4 (SD) to 81.5 +/- 14.2 one year after surgery (p<0.001). Although there was no correlation between the increase in visual quality of life and visual acuity, the increase in VFQ-25 could be well predicted: low visual acuity and significant impairment on VFQ-25 testing preoperatively made patients most likely to benefit from macular hole surgery. A relatively high retinal thickness measurement at the hole border measured on OCT further increases the predictive value. CONCLUSION: Macular hole surgery is associated with an increase in visual quality of life despite good visual acuity of the fellow eye. Preoperative visual acuity, VFQ-25 value and partly OCT may help to predict the increase in patients' vision related quality of life after surgery. PMID- 17077118 TI - Multifocal visual-evoked potential in unilateral compressive optic neuropathy. AB - AIM: To evaluate the effects of unilateral compressive optic neuropathy on amplitude and latency of multifocal visual evoked potentials (mfVEPs). METHODS: Static automated perimetry and mfVEP recordings were obtained from six patients with presumed meningiomas affecting one optic nerve. Monocular and interocular amplitude and latency analyses were performed and compared with normal control subjects. RESULTS: The change in the mfVEP amplitude agreed with visual field findings with regard to topography and severity of deviation from normal. The delay in recordable responses from affected eyes ranged from 7.6 to 20.7 ms (interocular analysis) and 7.9 to 13.9 ms (monocular analysis). CONCLUSIONS: Compressive optic neuropathy decreases the amplitude and increases the latency of the mfVEP. The changes in latency were similar to those seen in optic neuritis but larger than those in ischaemic optic neuropathy and glaucoma. PMID- 17077119 TI - Surgical and visual outcomes following exchange of opacified Hydroview intraocular lenses. AB - AIM: To report the clinical and surgical outcomes following exchange of opacified Hydroview intraocular lenses (IOLs), and to relate the final visual and anatomic results to clinical and surgical variables. METHODS: This is a prospective study of seventy-three eyes that underwent exchange of opacified Hydroview IOLs in Waterford Regional Hospital, Ireland. Preoperative, intraoperative and postoperative details were recorded. RESULTS: This study comprised 73 eyes of 71 consecutive patients undergoing IOL exchange, performed at mean (+/-SD) intervals of 36.64 (+/-9.9) months following the primary cataract surgery. The mean (+/-SE) follow-up following the exchange procedure was 13 (+/-1) months (range: 1-45 months). The secondary IOL was placed in the capsular bag, in the sulcus, and in the anterior chamber in 22 (30.1%), 24 (32.9%) and 27 (37%) cases, respectively. The IOL exchange procedure was uneventful in 36 eyes (49.3%), whereas intraoperative events such as posterior capsule rupture, vitreous loss and zonular dehiscence were seen in the remainder (50.7%). Following the IOL exchange procedure, a significant improvement in best corrected visual acuity (BCVA) was noted at one and at three months, and at the final visit (Wilcoxon signed ranks test: p<0.001, p = 0.006, and p<0.001, respectively). Following exclusion of eyes with visually consequential ocular comorbidity, a better final BCVA was noted among those eyes where the secondary IOL was placed in the capsular bag or in the sulcus when compared with placement of the secondary IOL in the anterior chamber (IOL in the bag or sulcus: 26 eyes (35.6%), median (IQR) final BCVA: 0.2 (0.10 0.40); IOL in the anterior chamber: 19 eyes (26.02%), median (IQR) final BCVA: 0.5 (0.20-0.60); Mann Whitney U Test: p = 0.004). CONCLUSION: IOL exchange is a technically challenging, but visually rewarding procedure. However, placement of the secondary IOL in the anterior chamber is associated with a poorer visual outcome when compared with placement of the secondary IOL in the sulcus or in the capsular bag. PMID- 17077120 TI - Pom1 kinase links division plane position to cell polarity by regulating Mid1p cortical distribution. AB - In fission yeast, Mid1p, a major determinant for division plane position, defines a medial cortical compartment where it recruits myosin II at the onset of mitosis to initiate contractile ring assembly. How Mid1p is restricted to the medial cortex is unknown. We report here that in a pom1 polarity mutant, which displays a monopolar growth pattern, Mid1p distribution expands towards the non-growing cell tip, uncoupling Mid1p localization from nuclear position. This accounts for the displacement of the contractile ring during mitosis. By contrast, Mid1p localization is normal in a bud6Delta strain, indicating that Mid1p misdistribution is not a general consequence of monopolar growth. We conclude that Pom1 kinase acts as a negative regulator of Mid1p distribution, excluding Mid1p from non-growing ends, whereas a Pom1-independent mechanism prevents Mid1p association with growing ends. Our work therefore provides evidence that cell polarity regulators influence the distribution of Mid1p, linking division plane position to cell polarity. PMID- 17077121 TI - Re-programming of newt cardiomyocytes is induced by tissue regeneration. AB - Newt hearts are able to repair substantial cardiac injuries without functional impairment, whereas mammalian hearts cannot regenerate. The cellular and molecular mechanisms that control the regenerative capacity of the newt heart are unknown. Here, we show that the ability of newt cardiomyocytes to regenerate cardiac injuries correlates with their ability to transdifferentiate into different cell types. Mechanical injury of the heart led to a severe reduction of sarcomeric proteins in the myocardium, indicating a partial de-differentiation of adult newt cardiomyocytes during regeneration. Newt cardiomyocytes implanted into regenerating limbs lost their cardiac phenotype and acquired skeletal muscle or chondrocyte fates. Reprogramming of cardiomyocytes depended on contact with the limb blastema because cardiomyocytes implanted into intact, non-regenerating limbs or cultured in vitro retained their original identity. We reason that signals from the limb blastema led to de-differentiation of cardiomyocytes, cell proliferation and re-differentiation into specialized cells and propose that the ability of cardiomyocytes to transdifferentiate into different cell types reflects the cellular program that enables heart regeneration. PMID- 17077122 TI - Genetic and molecular interactions of the Erv41p-Erv46p complex involved in transport between the endoplasmic reticulum and Golgi complex. AB - Erv41p and Erv46p are integral membrane proteins conserved across species. They were originally identified as abundant constituents of COPII-coated vesicles, and form a complex which cycles between the endoplasmic reticulum and Golgi complex. Yeast strains lacking these proteins are viable but display subtle secretory phenotypes. In order to obtain information about possible biological roles of this protein complex in endoplasmic reticulum to Golgi transport, we employed the Synthetic Genetic Array approach to screen for synthetic genetic interactions with the erv46 null mutation. We identified synthetic interactions with vma12, vma21, vma22 and vps1 deletion mutations. The vma21Delta mutation exacerbates transport defects caused by the erv46Delta mutation. Unexpectedly, yeast strains lacking Vma21p fail to sort the endoplasmic reticulum to Golgi v-SNARE, Bos1p, efficiently into COPII vesicles, yet these vesicles are fully fusion competent. In addition, we set out to identify, by a biochemical approach, proteins interacting with the Erv41p-Erv46p complex. We report a strong interaction between the Erv41p-Erv46p complex and endoplasmic reticulum glucosidase II. Strains lacking a cycling Erv41p-Erv46p complex display a mild glycoprotein processing defect. PMID- 17077123 TI - Influx of extracellular Ca2+ is necessary for electrotaxis in Dictyostelium. AB - Intracellular free Ca2+ ([Ca2+](i)) is a pivotal signalling element in cell migration and is thought to be required for chemotaxis of Dictyostelium. Ca2+ signalling may also be important for electrotaxis. However this suggestion has been controversial. We show that electric fields direct Dictyostelium cells to migrate cathodally and increase [Ca2+](i) in Dictyostelium cells, as determined by Fluo-3 AM imaging and (45)Ca2+ uptake. Omission of extracellular Ca2+([Ca2+](e)) and incubation with EGTA abolished the electric-field-stimulated [Ca2+](i) rise and directional cell migration. This suggests a requirement for [Ca2+](e) in the electrotactic response. Deletion of iplA, a gene responsible for chemoattractant-induced [Ca2+](i) increase, had only a minor effect on the electric-field-induced [Ca2+](i) rise. Moreover, iplA-null Dictyostelium cells showed the same electrotactic response as wild-type cells. Therefore, iplA independent Ca2+ influx is necessary for electrotactic cell migration. These results suggest that the [Ca2+](i) regulatory mechanisms induced by electric fields are different from those induced by cAMP and folic acid in Dictyostelium cells. Different roles of the iplA gene in chemoattractant-induced and electrically induced Ca2+ signalling, and different effects of [Ca2+](i) elevation on chemotaxis and electrotaxis indicate that the chemoattractant and electric cues activate distinctive initial signalling elements. PMID- 17077124 TI - Multiple domains in Siz SUMO ligases contribute to substrate selectivity. AB - Saccharomyces cerevisiae contains two Siz/PIAS SUMO E3 ligases, Siz1 and Siz2/Nfi1, and one other known ligase, Mms21. Although ubiquitin ligases are highly substrate-specific, the degree to which SUMO ligases target distinct sets of substrates is unknown. Here we show that although Siz1 and Siz2 each have unique substrates in vivo, sumoylation of many substrates can be stimulated by either protein. Furthermore, in the absence of both Siz proteins, many of the same substrates are still sumoylated at low levels. Some of this residual sumoylation depends on MMS21. Siz1 targets its unique substrates through at least two distinct domains. Sumoylation of PCNA (proliferating cell nuclear antigen) and the splicing factor Prp45 requires part of the N-terminal region of Siz1, the ;PINIT' domain, whereas sumoylation of the bud neck-associated septin proteins Cdc3, Cdc11 and Shs1/Sep7 requires the C-terminal domain of Siz1, which is also sufficient for cell cycle-dependent localization of Siz1 to the bud neck. Remarkably, the non-sumoylated septins Cdc10 and Cdc12 also undergo Siz1 dependent sumoylation if they are fused to the short PsiKXE SUMO attachment-site sequence. Collectively, these results suggest that local concentration of the E3, rather than a single direct interaction with the substrate polypeptide, is the major factor in substrate selectivity by Siz proteins. PMID- 17077125 TI - Macropinocytosis: regulated coordination of endocytic and exocytic membrane traffic events. AB - Macropinocytosis, a form of bulk uptake of fluid and solid cargo into cytoplasmic vacuoles, called macropinosomes, has been studied mostly in relation to antigen presentation. Early membrane traffic events occurring in this process are, however, largely unknown. Using human dendritic cells we show that a marked increase in the rate of macropinocytosis occurs a few minutes after application of two markers (small latex beads or dextran), depends on a slow intracellular Ca2+ concentration ([Ca2+](i)) rise that precedes the PI3K-dependent step, and is preceded and accompanied by exocytosis of enlargeosomes compensating in part for the macropinocytic plasma membrane internalization. Unexpectedly, macropinosomes themselves, which share markers with endosomes, undergo Ca2+ -dependent exocytosis so that, after approximately 20 minutes of continuous bead or dextran uptake, an equilibrium is reached preventing cells from overloading themselves with the organelles. Large [Ca2+](i) increases induced by ionomycin trigger rapid (<1 minute) exocytic regurgitation of all macropinosomes, whereas endosomes remain apparently unaffected. We conclude that, in dendritic cells, the rate of macropinocytosis is not constant but increases in a regulated fashion, as previously shown in other cell types. Moreover, macropinosomes are not simple containers that funnel cargo to an endocytic pathway, but unique organelles, distinct from endosomes by their competence for regulated exocytosis and other membrane properties. PMID- 17077126 TI - ARAP2 effects on the actin cytoskeleton are dependent on Arf6-specific GTPase activating-protein activity and binding to RhoA-GTP. AB - ARAP2 is a protein that contains both ArfGAP and RhoGAP domains. We found that it is a phosphatidylinositol (3,4,5)-trisphosphate-dependent Arf6 GAP that binds RhoA-GTP but lacks RhoGAP activity. In agreement with the hypothesis that ARAP2 mediates effects of RhoA, endogenous ARAP2 associated with focal adhesions (FAs) and reduction of ARAP2 expression, by RNAi, resulted in fewer FAs and actin stress fibers (SFs). In cells with reduced levels of endogenous ARAP2, FAs and SFs could be restored with wild-type recombinant ARAP2 but not mutants lacking ArfGAP or Rho-binding activity. Constitutively active Arf6 also caused a loss of SFs. The Rho effector ROKalpha was ineffective in restoring FAs. Conversely, overexpression of ARAP2 did not restore SFs in cells treated with a ROK inhibitor but induced punctate accumulations of paxillin. We conclude that ARAP2 is an Arf6GAP that functions downstream of RhoA to regulate focal adhesion dynamics. PMID- 17077127 TI - Kinesin 6 family member Subito participates in mitotic spindle assembly and interacts with mitotic regulators. AB - Drosophila Subito is a kinesin 6 family member and ortholog of mitotic kinesin like protein (MKLP2) in mammalian cells. Based on the previously established requirement for Subito in meiotic spindle formation and for MKLP2 in cytokinesis, we investigated the function of Subito in mitosis. During metaphase, Subito localized to microtubules at the center of the mitotic spindle, probably interpolar microtubules that originate at the poles and overlap in antiparallel orientation. Consistent with this localization pattern, subito mutants improperly assembled microtubules at metaphase, causing activation of the spindle assembly checkpoint and lagging chromosomes at anaphase. These results are the first demonstration of a kinesin 6 family member with a function in mitotic spindle assembly, possibly involving the interpolar microtubules. However, the role of Subito during mitotic anaphase resembles other kinesin 6 family members. Subito localizes to the spindle midzone at anaphase and is required for the localization of Polo, Incenp and Aurora B. Genetic evidence suggested that the effects of subito mutants are attenuated as a result of redundant mechanisms for spindle assembly and cytokinesis. For example, subito double mutants with ncd, polo, Aurora B or Incenp mutations were synthetic lethal with severe defects in microtubule organization. PMID- 17077128 TI - Studies of the common DIO2 Thr92Ala polymorphism and metabolic phenotypes in 7342 Danish white subjects. AB - CONTEXT: The type 2 iodothyronine deiodinase (D2) catalyzes the conversion of T(4) to the active form of thyroid hormone, which is a critical regulator of thermogenesis and glucose metabolism. A Thr92Ala polymorphism in the gene encoding D2 (DIO2) has been reported to associate with insulin resistance. OBJECTIVE: The aim of the present study was to assess the impact of the DIO2 Thr92Ala variant on type 2 diabetes (T2D), obesity, and related quantitative metabolic traits including measures of insulin resistance. Because DIO2 is activated through a beta-adrenergic receptor-dependent pathway, we further hypothesized that variation in the ADRB genes interacts with DIO2 Thr92Ala variant to influence metabolic traits. DESIGN AND PATIENTS: The DIO2 polymorphism was genotyped in a total of 7342 white subjects including 1405 T2D patients. RESULTS: We detected no significant association of the DIO2 Thr92Ala polymorphism with T2D or obesity. We observed nominal significant associations of genotype with increased area under the serum insulin curve during an oral glucose tolerance test (P = 0.03) and elevated fasting plasma glucose (P = 0.02) in homozygous Ala92 allele carriers, the latter strengthened by epistasis with the ADRB2 Gly16Arg variant in a double recessive model (P = 0.004). However, after permutation procedure, performed to correct for multiple hypothesis testing, the associations did not reach study-wide significance. CONCLUSIONS: The DIO2 Thr92Ala variant does not confer an increased risk of T2D, obesity, or insulin resistance. PMID- 17077129 TI - Sodium/iodide symporter (NIS) gene expression is the limiting step for the onset of thyroid function in the human fetus. AB - CONTEXT: Terminal differentiation of the human thyroid is characterized by the onset of follicle formation and thyroid hormone synthesis at 11 gestational weeks (GW). OBJECTIVE: This study aimed to investigate the ontogeny of thyroglobulin (Tg), thyroid peroxidase (TPO), sodium/iodide symporter (NIS), pendrin (PDS), dual oxidase 2 (DUOX2), thyroid-stimulating hormone receptor (TSHR), and thyroid transcription factor 1 (TITF1), forkhead box E1 (FOXE1), and paired box gene 8 (PAX8) in the developing human thyroid. DESIGN: Thyroid tissues from human embryos and fetuses (7-33 GW; n = 45) were analyzed by quantitative PCR to monitor mRNA expression for each gene and by immunohistochemistry to determine the cellular distribution of TITF1, TSHR, Tg, TPO, NIS, and the onset of T4 production. A broken line regression model was fitted for each gene to compare the loglinear increase in expression before and after the onset of T4 synthesis. RESULTS: TITF1, FOXE1, PAX8, TSHR, and DUOX2 were stably expressed from 7 to 33 GW. Tg, TPO, and PDS expression was detectable as early as 7 GW and was correlated with gestational age (all, P < 0.01), and the slope of the regression line was significantly different before and after the onset of T4 synthesis at 11 GW (all, P < 0.01). NIS expression appeared last and showed the highest fit by the broken line regression model of all genes (correlation age P < 0.0001, broken line regression P < 0.0001). Immunohistochemical studies detected TITF1, TSHR, and Tg in unpolarized thyrocytes before follicle formation. T(4) and NIS labeling were only found in developing follicles from 11 GW on. CONCLUSION: These results imply a key role of NIS for the onset of human thyroid function. PMID- 17077130 TI - Prognostic value of the sixth edition AJCC/UICC TNM classification for differentiated thyroid carcinoma with extrathyroid extension. AB - CONTEXT: The prognostic value of the sixth edition AJCC/UICC TNM classification is currently unclear. OBJECTIVE: The aim was to evaluate the prognostic value of the sixth edition. DESIGN AND PATIENTS: We retrospectively assessed 354 primary differentiated thyroid carcinomas (77 men and 277 women; age, 51.2 yr; follow-up, 107.6 months) between 1964 and 2003. Sixty percent of patients underwent lobectomy, 40% underwent subtotal/total thyroidectomy, and only 2% were given radioiodine. There were 153, 104, 86, and 11 patients in fifth stages I, II, III, and IV, and 175, 76, 14, 68, 10, and 11 patients in sixth stages I, II, III, IVA, IVB, and IVC, respectively. RESULTS: New T1-3 had no significant influence. In Cox proportional hazard analysis, T4a and T4b were significantly related to disease-specific survival (DSS). We separately analyzed 68 patients (age 45 yr or older) with extrathyroid extension. These T4 (fifth) tumors were reclassified as 6 T3, 52 T4a, and 10 T4b tumors. The 10-yr DSS rates were 100, 69.3, and 10.0% for T3, T4a, and T4b, respectively. T4b exhibited worse prognoses compared with T4a (P < 0.0001; hazard ratio, 10.1; 95% confidence interval, 4.1-25.3). Stages I and II in both editions achieved favorable prognoses. The 10-yr DSS rates were 67.0 and 27.3% in fifth stages III and IV, and 100, 74.5, 10.0, and 27.3% in sixth stages III, IVA, IVB, and IVC, respectively. DSS curves differed significantly between all sixth TNM stages (P < 0.0001). CONCLUSION: The sixth edition more accurately predicts different outcomes according to the revised criteria for the degree of extrathyroid extension. PMID- 17077131 TI - Gender, body weight, disease activity, and previous radiotherapy influence the response to pegvisomant. AB - CONTEXT/OBJECTIVE: To effectively normalize IGF-I in patients with acromegaly, various covariates may affect dosing and plasma concentrations of pegvisomant. We assessed whether sex, age, weight, and previous radiotherapy influence dosing of pegvisomant in patients with active disease. DESIGN: Data from 69 men and 49 women participating in multicenter, open-label trials of pegvisomant were retrospectively evaluated using multiple regression techniques. Sixty-nine subjects (39 men, 30 women) had undergone external beam pituitary radiotherapy. Serum IGF-I was at least 30% above age-related upper limit of normal in all patients at study entry. After a loading dose of pegvisomant (80 mg), patients were commenced on 10 mg/d. Pegvisomant dose was adjusted by 5 mg every eighth week until serum IGF-I was normalized. RESULTS: At baseline, men had significantly higher mean serum IGF-I levels than women despite similar GH levels. After treatment with pegvisomant, IGF-I levels were similar in men and women. A significant correlation between baseline GH, IGF-I, body weight, and the dose of pegvisomant required to normalize serum IGF-I was observed (all P < 0.001). Women required an average of 0.04 mg/kg more pegvisomant than men and a mean weight-corrected dose of 19.2 mg/d to normalize serum IGF-I [14.5 mg/d (men); P < 0.001]. Patients treated with radiotherapy required less pegvisomant to normalize serum IGF-I despite similar baseline GH/IGF-I levels (15.2 vs. 18.5 mg/d for no previous radiotherapy; P = 0.002). CONCLUSIONS: Sex, body weight, previous radiotherapy, and baseline GH/IGF-I influence the dose of pegvisomant required to normalize serum IGF-I in patients with active acromegaly. PMID- 17077132 TI - Smoking is associated with lower bone mineral density and reduced cortical thickness in young men. AB - CONTEXT: Smoking has previously been associated with reduced areal bone mineral density (aBMD) in elderly subjects, but the association remains controversial in adolescents. OBJECTIVE: The aim of this study was to determine whether smoking was associated with aBMD or volumetric BMD (vBMD) and bone size in young men. DESIGN AND SETTING: aBMD was measured using dual x-ray absorptiometry. vBMD and bone size were measured using peripheral quantitative computerized tomography (pQCT). Smoking habits were assessed using questionnaires. Levels of sex steroids, PTH, and 25-OH-vitamin D were measured in serum. PARTICIPANTS: The population-based Gothenburg Osteoporosis and Obesity Determinants (GOOD) study includes 1068 young men, age 18.9 +/- 0.6 yr (mean +/- SD). MAIN OUTCOME MEASURE: The main outcome measure was smoking as predictor of bone parameters and serum sex hormone levels. RESULTS: Of the study subjects, 8.7% smoked daily. Bone parameters were compared between smokers and nonsmokers. Smokers had significantly lower aBMD (dual x-ray absorptiometry) of the total body (crude: 2.1%; adjusted for age, height, weight, calcium intake, and physical activity: 1.8%), lumbar spine (crude: -4.3%; adjusted: -3.3%), and trochanter (crude: 6.6%; adjusted: -5.0%) than nonsmokers. Using peripheral quantitative computerized tomography, we found that smokers had lower cortical thickness of both the radius (crude: -2.8%; adjusted: -2.9%) and tibia (crude: -4.5%; adjusted: -4.0%) than the nonsmokers, whereas no difference was seen for cortical vBMD. Smokers had higher serum levels of total and free testosterone and lower 25 OH-vitamin D than nonsmokers. Adjustment for testosterone and/or 25-OH-vitamin D levels did not alter the associations between smoking and bone parameters. CONCLUSIONS: We demonstrate that smoking was associated with lower aBMD and reduced cortical thickness in young men. PMID- 17077133 TI - Monitoring thyroglobulin in a sensitive immunoassay has comparable sensitivity to recombinant human tsh-stimulated thyroglobulin in follow-up of thyroid cancer patients. AB - CONTEXT: Most thyroglobulin (Tg) assays have a sensitivity of 0.5-1 ng/ml. A minority of patients with undetectable T4-suppressed Tg levels have a recombinant human TSH (rhTSH)-stimulated Tg above 2 ng/ml and identifiable residual disease. OBJECTIVE: The objective was to determine whether a Tg assay with improved sensitivity could eliminate the need for rhTSH stimulation when baseline Tg is below 0.1 ng/ml. DESIGN: A retrospective study of two academic endocrine practices was conducted. POPULATION: A total of 194 patients undergoing rhTSH stimulation participated in the study. RESULTS: Of the 80 patients with Tg below 0.1 ng/ml, two (2.5%) had rhTSH-stimulated Tg above 2 ng/ml. One other patient with stimulation to 0.3 ng/ml and negative 123I scan had an ultrasound-detected malignant lymph node resected. None had 131I/123I imaging after rhTSH stimulation suggestive of local recurrence or distant metastasis. If T4-suppressed Tg was 0.1 0.5 or 0.6-2.0 ng/ml, rhTSH Tg was above 2 ng/ml in 24.2 and 82.4%, respectively. CONCLUSIONS: Patients with differentiated thyroid carcinoma and a T4-suppressed serum Tg below 0.1 ng/ml rarely have a rhTSH-stimulated Tg above 2 ng/ml, and none of these patients had 131I or 123I imaging after rhTSH stimulation suggestive of local recurrence or distant metastasis. We recommend monitoring such patients with a T4-suppressed Tg level and periodic neck ultrasonography. An increase in T4-suppressed serum Tg to a detectable level or the appearance of abnormal lymph nodes by physical or ultrasound exam should prompt further investigation. PMID- 17077134 TI - A brief exposure to moderate passive smoke increases metabolism and thyroid hormone secretion. AB - CONTEXT: Active smoking influences normal metabolic status and thyroid function. OBJECTIVE: The objective was to assess experimentally the effects of 1 h of moderate passive smoking in a controlled simulated bar/restaurant environment on the metabolism and thyroid hormone levels in healthy nonsmokers. PARTICIPANTS: Eighteen (nine females, nine males) healthy individuals (mean +/- sd: age, 25.3 +/- 3.1 yr; height, 174.0 +/- 10.1 cm; weight, 65.2 +/- 13.7 kg) participated in the study. DESIGN: In repeated-measures randomized blocks, participants visited the laboratory on 2 consecutive days. In the experimental condition, they were exposed to 1 h of moderate passive smoking at a carbon monoxide concentration of 23 +/- 1 ppm in an environmental chamber, whereas in the control condition participants remained in the same chamber for 1 h breathing normal atmospheric air. MAIN OUTCOME MEASURES: In both conditions, cotinine serum and urine levels, resting energy expenditure (REE), as well as concentration of T3, free T4, and TSH were assessed before participants entered the chamber and immediately after their exit. Heart rate and blood pressure were tested in 10-min intervals during all REE assessments. RESULTS: The mean +/- sd difference of serum and urine cotinine levels (-0.27 +/- 3.94 vs. 14.01 +/- 6.54 and 0.05 +/- 2.07 vs. 7.23 +/- 3.75, respectively), REE (6.73 +/- 98.06 vs. 80.58 +/- 120.91) as well as T3 and free T4 (0.05 +/- 0.11 vs. 0.13 +/- 0.12 and 0.02 +/- 0.15 vs. 0.22 +/- 0.20) were increased in the experimental compared with the control condition at baseline and follow-up (P < 0.05). No statistically significant variation was observed in the mean difference of the remaining parameters (P > 0.05). Serum and urine cotinine values were linearly associated with REE (P < 0.05). CONCLUSION: One hour of passive smoking at bar/restaurant levels is accompanied by significant increases in metabolism and thyroid hormone levels. PMID- 17077135 TI - Parturition itself is the basis for fetal adrenal involution. AB - CONTEXT: Newborn infants show a postnatal decline in androgen levels as the fetal adrenal glands involute. HYPOTHESIS: Placental factors up-regulate dehydroepiandrosterone sulfate (DHEA-S) generation. Hence, regardless of age, parturition will result in fetal adrenal involution and decline in DHEA-S levels. SUBJECTS AND METHODS: Premature neonates (n = 30) with gestational age 26-35 wk were studied. Adrenal volume by ultrasonography and serum DHEA-S, cortisol, and androstendione levels were followed weekly between d 1 and 28 of life. RESULTS: Serum DHEA-S was high on d 1 of life, declining rapidly regardless of gestational age during the first week of life (P < 0.001), and serum androstenedione and cortisol levels followed a similar pattern. Androstenedione levels showed a rise as of d 21 of life in boys but not in girls. The adrenals decreased in ultrasonographic volume from d 1 to 14 of life (P < 0.001), regardless of gestational age. CONCLUSIONS: Involution of the adrenal is faster than previously reported and, regardless of gestational age, occurs within the first week of life in terms of hormone secretion and within 2 wk in adrenal size. Involution involves a decline in DHEA-S but also in androstenedione and cortisol secretion, with a change in enzymatic activity. Males and females differ in their androstenedione levels and enzymatic activity. Parturition itself is the basis for fetal adrenal involution, supporting a key role for placental factors in maintaining the fetal adrenal and generating adrenal androgens. PMID- 17077136 TI - Methods and protocols for prediction of immunogenic epitopes. AB - T-cell recognition of peptide/major histocompatibility complex (MHC) is a prerequisite for cellular immunity. Recently, there has been an influx of bioinformatics tools to facilitate the identification of T-cell epitopes to specific MHC alleles. This article examines existing computational strategies for the study of peptide/MHC interactions. The most important bioinformatics tools and methods with relevance to the study of peptide/MHC interactions have been reviewed. We have also provided guidelines for predicting antigenic peptides based on the availability of existing experimental data. PMID- 17077137 TI - Enrichment analysis in high-throughput genomics - accounting for dependency in the NULL. AB - Translating the overwhelming amount of data generated in high-throughput genomics experiments into biologically meaningful evidence, which may for example point to a series of biomarkers or hint at a relevant pathway, is a matter of great interest in bioinformatics these days. Genes showing similar experimental profiles, it is hypothesized, share biological mechanisms that if understood could provide clues to the molecular processes leading to pathological events. It is the topic of further study to learn if or how a priori information about the known genes may serve to explain coexpression. One popular method of knowledge discovery in high-throughput genomics experiments, enrichment analysis (EA), seeks to infer if an interesting collection of genes is 'enriched' for a Consortium particular set of a priori Gene Ontology Consortium (GO) classes. For the purposes of statistical testing, the conventional methods offered in EA software implicitly assume independence between the GO classes. Genes may be annotated for more than one biological classification, and therefore the resulting test statistics of enrichment between GO classes can be highly dependent if the overlapping gene sets are relatively large. There is a need to formally determine if conventional EA results are robust to the independence assumption. We derive the exact null distribution for testing enrichment of GO classes by relaxing the independence assumption using well-known statistical theory. In applications with publicly available data sets, our test results are similar to the conventional approach which assumes independence. We argue that the independence assumption is not detrimental. PMID- 17077138 TI - Evolving research trends in bioinformatics. AB - The cross-disciplinary nature of bioinformatics entails co-evolution with other biomedical disciplines, whereby some bioinformatics applications become popular in certain disciplines and, in turn, these disciplines influence the focus of future bioinformatics development efforts. We observe here that the growth of computational approaches within various biomedical disciplines is not merely a reflection of a general extended usage of computers and the Internet, but due to the production of useful bioinformatics databases and methods for the rest of the biomedical scientific community. We have used the abstracts stored both in the MEDLINE database of biomedical literature and in NIH-funded project grants, to quantify two effects. First, we examine the biomedical literature as a whole and find that the use of computational methods has become increasingly prevalent across biomedical disciplines over the past three decades, while use of databases and the Internet have been rapidly increasing over the past decade. Second, we study the recent trends in the use of bioinformatics topics. We observe that molecular sequence databases are a widely adopted contribution in biomedicine from the field of bioinformatics, and that microarray analysis is one of the major new topics engaged by the bioinformatics community. Via this analysis, we were able to identify areas of rapid growth in the use of informatics to aid in curriculum planning, development of computational infrastructure and strategies for workforce education and funding. PMID- 17077139 TI - Structure of a heparin-dependent complex of Hedgehog and Ihog. AB - Hedgehog (Hh) signaling molecules mediate key tissue-patterning events during animal development, and inappropriate activation of Hh signaling in adults has been associated with human cancers. Recently, a conserved family of type I integral membrane proteins required for normal response to the Hh signal was discovered. One member of this family, Ihog (interference hedgehog), functions upstream or at the level of Patched (Ptc), but how Ihog participates in Hh signaling remains unclear. Here, we show that heparin binding induces Ihog dimerization and is required to mediate high-affinity interactions between Ihog and Hh. We also present crystal structures of a Hh-binding fragment of Ihog, both alone and complexed with Hh. Heparin is not well ordered in these structures, but a basic cleft in the first FNIII domain of Ihog (IhogFn1) is shown by mutagenesis to mediate heparin binding. These results establish that Hh directly binds Ihog and provide the first demonstration of a specific role for heparin in Hh responsiveness. PMID- 17077140 TI - Expression of TEL-JAK2 in primary human hematopoietic cells drives erythropoietin independent erythropoiesis and induces myelofibrosis in vivo. AB - Activation of JAK2 by chromosomal translocation or point mutation is a recurrent event in hematopoietic malignancies, including acute leukemias and myeloproliferative disorders. Although the effects of activated JAK2 signaling have been examined in cell lines and murine models, the functional consequences of deregulated JAK2 in the context of human hematopoietic cells are currently unknown. Here we report that expression of TEL-JAK2, a constitutively active variant of the JAK2 kinase, in lineage-depleted human umbilical cord blood cells results in erythropoietin-independent erythroid differentiation in vitro and induces the rapid development of myelofibrosis in an in vivo NOD/SCID xenotransplantation assay. These studies provide functional evidence that activated JAK2 signaling in primitive human hematopoietic cells is sufficient to drive key processes implicated in the pathophysiology of polycythemia vera and idiopathic myelofibrosis. Furthermore, they describe an in vivo model of myelofibrosis initiated with primary cells, highlighting the utility of the NOD/SCID xenotransplant system for the development of experimental models of human hematopoietic malignancies. PMID- 17077141 TI - Profile of F. Stuart Chapin III. PMID- 17077142 TI - Activation of estrogen receptor alpha increases and estrogen receptor beta decreases apolipoprotein E expression in hippocampus in vitro and in vivo. AB - Previous evidence indicates that, in carriers of apolipoprotein E4 (ApoE4), estrogen therapy increased the risk of late-onset Alzheimer's disease (AD), whereas in individuals carrying ApoE2/3, estrogen therapy reduced the risk of AD [Cauley JA, Zmuda JM, Yaffe K, Kuller LH, Ferrell RE, Wisniewski SR, Cummings SR (1999) J Bone Miner Res 14:1175-1181; Yaffe K, Haan M, Byers A, Tangen C, Kuller L (2000) Neurology 54:1949-1954]. Estrogen mechanisms of action are mediated by two estrogen receptors (ERs), ERalpha and ERbeta. In this study, we determined the relationship between ER subtype and estrogen regulation of ApoE expression in HT-22 cells ectopically transfected with ERalpha or ERbeta, in primary cultured rat hippocampal neurons in vitro and in rat hippocampus in vivo by both molecular biological and pharmacological analyses. Results of these analyses demonstrated that activation of ERalpha either by 17beta-estradiol or a specific-agonist, propylpyrazole triol, up-regulated ApoE mRNA and protein expression. In contrast, the ERbeta-selective agonist, diarylpropionitrile, down-regulated ApoE mRNA and protein expression. These results demonstrate that, in vitro and in vivo, ApoE expression can be differentially regulated depending on activation of ER subtypes. These data suggest that use of ER-selective ligands could provide therapeutic benefit to reduce the risk of AD by increasing ApoE expression in ApoE2/3 allele carriers and decreasing ApoE expression in ApoE4 allele carriers. PMID- 17077143 TI - NMDA receptors promote survival in somatosensory relay nuclei by inhibiting Bax dependent developmental cell death. AB - Naturally occurring cell death is a universal feature of developing nervous systems that plays an essential role in determining adult brain function. Yet little is known about the decisions that select a subset of CNS neurons for survival and cause others to die. We report that postnatal day 0 NMDA receptor subunit 1 (NMDAR1) knockout mice display an approximately 2-fold increase in cell death in the brainstem trigeminal complex (BSTC), including all four nuclei that receive somatosensory inputs from the face (principalis, oralis, interpolaris, and caudalis). Treatment with the NMDA receptor antagonist dizocilpine maleate (MK-801) for 24 h before birth also caused an increase in cell death that reached statistical significance in two of the four nuclei (oralis and interpolaris). The neonatal sensitivity to NMDA receptor hypofunction in the BSTC, and in its main thalamic target, the ventrobasal nucleus (VB), coincides with the peak of naturally occurring cell death and trigeminothalamic synaptogenesis. At embryonic day 17.5, before the onset of these events, NMDAR1 knockout does not affect cell survival in either the BSTC or the VB. Immunostaining for active caspase-3 and the neuronal marker Hu specifically confirms the presence of dying neurons in the BSTC and the VB of NMDAR1 knockout neonates. Finally, genetic deletion of Bax rescues these structures from the requirement for NMDA receptors to limit naturally occurring cell death. Taken together, the results indicate that NMDA receptors play a survival role for somatosensory relay neurons during synaptogenesis by inhibiting Bax-dependent developmental cell death. PMID- 17077144 TI - Essential role of RGS-PX1/sorting nexin 13 in mouse development and regulation of endocytosis dynamics. AB - RGS-PX1 (also known as sorting nexin 13) is a member of both the regulator of G protein signaling (RGS) and sorting nexin (SNX) protein families. Biochemical and cell culture studies have shown that RGS-PX1/SNX13 attenuates Galphas-mediated signaling through its RGS domain and regulates endocytic trafficking and degradation of the epidermal growth factor receptor. To understand the functions of RGS-PX1/SNX13 in vivo, we generated mice carrying targeted mutations of Snx13 and found that systemic Snx13-null mice were embryonic lethal around midgestation. Snx13-null embryos had significant overall growth retardation and defects in neural tube closure, blood vessel formation, and the formation of the placental labyrinthine layer. Moreover, the Snx13-null visceral yolk sac endoderm cells showed dramatic changes in the organization of endocytic compartments, abundant autophagic vacuoles, and abnormal localization of several endocytic markers, including megalin, a receptor for nutrients and proteins; ARH, a coat protein that binds megalin; LAMP2; and LC3. These changes suggest that Snx13-null embryos are defective in nutrient uptake and transport, which may contribute to the other developmental abnormalities observed. Taken together, our findings demonstrate an essential role for RGS-PX1/SNX13 in mouse development and provide previously undescribed insights into its cellular function in the regulation of endocytosis dynamics. PMID- 17077145 TI - Quantum dot/peptide-MHC biosensors reveal strong CD8-dependent cooperation between self and viral antigens that augment the T cell response. AB - Cytotoxic T lymphocytes (CTL) can respond to a few viral peptide-MHC-I (pMHC-I) complexes among a myriad of virus-unrelated endogenous self pMHC-I complexes displayed on virus-infected cells. To elucidate the molecular recognition events on live CTL, we have utilized a self-assembled biosensor composed of semiconductor nanocrystals, quantum dots, carrying a controlled number of virus derived (cognate) and other (noncognate) pMHC-I complexes and examined their recognition by antigen-specific T cell receptor (TCR) on anti-virus CD8(+) T cells. The unique architecture of nanoscale quantum dot/pMHC-I conjugates revealed that unexpectedly strong multivalent CD8-MHC-I interactions underlie the cooperative contribution of noncognate pMHC-I to the recognition of cognate pMHC I by TCR to augment T cell responses. The cooperative, CD8-dependent spread of signal from a few productively engaged TCR to many other TCR can explain the remarkable ability of CTL to respond to virus-infected cells that present few cognate pMHC-I complexes. PMID- 17077146 TI - Implications of the quaternary twist allosteric model for the physiology and pathology of nicotinic acetylcholine receptors. AB - Nicotinic acetylcholine receptors (nAChR) are pentameric ligand-gated ion channels composed of subunits that consist of an extracellular domain that carries the ligand-binding site and a distinct ion-pore domain. Signal transduction results from the allosteric coupling between the two domains: the distance from the binding site to the gate of the pore domain is 50 A. Normal mode analysis with a C(alpha) Gaussian network of a new structural model of the neuronal alpha7 nAChR showed that the lowest mode involves a global quaternary twist motion that opens the ion pore. A molecular probe analysis, in which the network is modified at each individual amino acid residue, demonstrated that the major effect is to change the frequency, but not the form, of the twist mode. The largest effects were observed for the ligand-binding site and the Cys-loop. Most (24/27) of spontaneous mutations known to cause congenital myasthenia and autosomal dominant nocturnal frontal lobe epilepsy are located either at the interface between subunits or, within a given subunit, at the interface between rigid blocks. These interfaces are modified significantly by the twist mode. The present analysis, thus, supports the quaternary twist model of the nAChR allosteric transition and provides a qualitative interpretation of the effect of the mutations responsible for several receptor pathologies. PMID- 17077147 TI - Targeting multiple kinase pathways in leukemic progenitors and stem cells is essential for improved treatment of Ph+ leukemia in mice. AB - It is generally believed that shutting down the kinase activity of BCR-ABL by imatinib will completely inhibit its functions, leading to inactivation of its downstream signaling pathways and cure of the disease. Imatinib is highly effective at treating human Philadelphia chromosome-positive (Ph(+)) chronic myeloid leukemia (CML) in chronic phase but not Ph(+) B cell acute lymphoblastic leukemia (B-ALL) and CML blast crisis. We find that SRC kinases activated by BCR ABL remain fully active in imatinib-treated mouse leukemic cells, suggesting that imatinib does not inactivate all BCR-ABL-activated signaling pathways. This SRC pathway is essential for leukemic cells to survive imatinib treatment and for CML transition to lymphoid blast crisis. Inhibition of both SRC and BCR-ABL kinase activities by dasatinib affords complete B-ALL remission. However, curing B-ALL and CML mice requires killing leukemic stem cells insensitive to both imatinib and dasatinib. Besides BCR-ABL and SRC kinases, stem cell pathways must be targeted for curative therapy of Ph(+) leukemia. PMID- 17077148 TI - Genetic basis for natural variation in seed vitamin E levels in Arabidopsis thaliana. AB - Vitamin E is an essential nutrient for humans and is obtained primarily from food, especially oil, derived from the seed of plants. Genes encoding the committed steps in vitamin E synthesis in plants (VTE, loci 1-5) have been isolated and used for tocopherol pathway engineering with various degrees of success. As a complement to such approaches we have used quantitative trait loci analysis with two sets of Arabidopsis thaliana recombinant inbred lines and have identified 14 QVE (quantitative vitamin E) loci affecting tocopherol content and composition in seeds. Five QVE intervals contain VTE loci that are likely QVE gene candidates. Nine QVE intervals do not contain VTE loci and therefore identify novel loci affecting seed tocopherol content and composition. Several near-isogenic lines containing introgressions of the accession with increased vitamin E levels were shown to confer significantly elevated tocopherol levels compared with the recurrent parent. Fine-mapping has narrowed QVE7 (a gamma tocopherol quantitative trait loci) to an 8.5-kb interval encompassing two genes. Understanding the basis of the QVE loci in Arabidopsis promises to provide insight into the regulation and/or metabolism of vitamin E in plants and has clear ramifications for improving the nutritional content of crops through marker assisted selection and metabolic engineering. PMID- 17077149 TI - Hagfish and lancelet fibrillar collagens reveal that type II collagen-based cartilage evolved in stem vertebrates. AB - The origin of vertebrates was defined by evolution of a skeleton; however, little is known about the developmental mechanisms responsible for this landmark evolutionary innovation. In jawed vertebrates, cartilage matrix consists predominantly of type II collagen (Col2alpha1), whereas that of jawless fishes has long been thought to be noncollagenous. We recently showed that Col2alpha1 is present in lamprey cartilage, indicating that type II collagen-based cartilage evolved earlier than previously recognized. Here, we investigate the origin of vertebrate cartilage, and we report that hagfishes, the sister group to lampreys, also have Col2alpha1-based cartilage, suggesting its presence in the common ancestor of crown-group vertebrates. We go on to show that lancelets, a sister group to vertebrates, possess an ancestral clade A fibrillar collagen (ColA) gene that is expressed in the notochord. Together, these results suggest that duplication and diversification of ColA genes at the chordate-vertebrate transition may underlie the evolutionary origin of vertebrate skeletal tissues. PMID- 17077150 TI - Akt1 deficiency affects neuronal morphology and predisposes to abnormalities in prefrontal cortex functioning. AB - There is accumulating evidence that AKT signaling plays a role in the pathogenesis of schizophrenia. We asked whether Akt1 deficiency in mice results in structural and functional abnormalities in prefrontal cortex (PFC). Exploratory transcriptional profiling revealed concerted alterations in the expression of PFC genes controlling synaptic function, neuronal development, myelination, and actin polymerization, and follow-up ultrastructural analysis identified consistent changes in the dendritic architecture of pyramidal neurons. Behavioral analysis indicated that Akt1-mutant mice have normal acquisition of a PFC-dependent cognitive task but abnormal working memory retention under neurochemical challenge of three distinct neurotransmitter systems. Thus, Akt1 deficiency creates a context permissive for gene-gene and gene-environment interactions that modulate PFC functioning and contribute to the disease risk associated with this locus, the severity of the clinical syndrome, or both. PMID- 17077151 TI - Wnt and TGF-beta signaling are required for the induction of an in vitro model of primitive streak formation using embryonic stem cells. AB - The establishment of the primitive streak and its derivative germ layers, mesoderm and endoderm, are prerequisite steps in the formation of many tissues. To model these developmental stages in vitro, an ES cell line was established that expresses CD4 from the foxa2 locus in addition to GFP from the brachyury locus. A GFP-Bry(+) population expressing variable levels of CD4-Foxa2 developed upon differentiation of this ES cell line. Analysis of gene-expression patterns and developmental potential revealed that the CD4-Foxa2(hi)GFP-Bry(+) population displays characteristics of the anterior primitive streak, whereas the CD4 Foxa2(lo)GFP-Bry(+) cells resemble the posterior streak. Using this model, we were able to demonstrate that Wnt and TGF-beta/nodal/activin signaling simultaneously were required for the generation of the CD4-Foxa2(+)GFP-Bry(+) population. Wnt or low levels of activin-induced a posterior primitive streak population, whereas high levels of activin resulted in an anterior streak fate. Finally, sustained activin signaling was found to stimulate endoderm commitment from the CD4-Foxa2(+)GFP-Bry(+) ES cell population. These findings demonstrate that the early developmental events involved in germ-layer induction in the embryo are recapitulated in the ES cell model and uncover insights into the signaling pathways involved in the establishment of mesoderm and endoderm. PMID- 17077152 TI - MyoD expression restores defective myogenic differentiation of human mesoangioblasts from inclusion-body myositis muscle. AB - Inflammatory myopathies (IM) are acquired diseases of skeletal muscle comprising dermatomyositis (DM), polymyositis (PM), and inclusion-body myositis (IBM). Immunosuppressive therapies, usually beneficial for DM and PM, are poorly effective in IBM. We report the isolation and characterization of mesoangioblasts, vessel-associated stem cells, from diagnostic muscle biopsies of IM. The number of cells isolated, proliferation rate and lifespan, markers expression, and ability to differentiate into smooth muscle do not differ among normal and IM mesoangioblasts. At variance with normal, DM and PM mesoangioblasts, cells isolated from IBM, fail to differentiate into skeletal myotubes. These data correlate with lack in connective tissue of IBM muscle of alkaline phosphatase (ALP)-positive cells, conversely dramatically increased in PM and DM. A myogenic inhibitory basic helix-loop-helix factor B3 is highly expressed in IBM mesoangioblasts. Indeed, silencing this gene or overexpressing MyoD rescues the myogenic defect of IBM mesoangioblasts, opening novel cell-based therapeutic strategies for this crippling disorder. PMID- 17077153 TI - muO-conotoxin MrVIB selectively blocks Nav1.8 sensory neuron specific sodium channels and chronic pain behavior without motor deficits. AB - The tetrodotoxin-resistant voltage-gated sodium channel (VGSC) Na(v)1.8 is expressed predominantly by damage-sensing primary afferent nerves and is important for the development and maintenance of persistent pain states. Here we demonstrate that muO-conotoxin MrVIB from Conus marmoreus displays substantial selectivity for Na(v)1.8 and inhibits pain behavior in models of persistent pain. In rat sensory neurons, submicromolar concentrations of MrVIB blocked tetrodotoxin-resistant current characteristic of Na(v)1.8 but not Na(v)1.9 or tetrodotoxin-sensitive VGSC currents. MrVIB blocked human Na(v)1.8 expressed in Xenopus oocytes with selectivity at least 10-fold greater than other VGSCs. In neuropathic and chronic inflammatory pain models, allodynia and hyperalgesia were both reduced by intrathecal infusion of MrVIB (0.03-3 nmol), whereas motor side effects occurred only at 30-fold higher doses. In contrast, the nonselective VGSC blocker lignocaine displayed no selectivity for allodynia and hyperalgesia versus motor side effects. The actions of MrVIB reveal that VGSC antagonists displaying selectivity toward Na(v)1.8 can alleviate chronic pain behavior with a greater therapeutic index than nonselective antagonists. PMID- 17077154 TI - Insertion events of CR1 retrotransposable elements elucidate the phylogenetic branching order in galliform birds. AB - Using standard phylogenetic methods, it can be hard to resolve the order in which speciation events took place when new lineages evolved in the distant past and within a short time frame. As an example, phylogenies of galliform birds (including well-known species such as chicken, turkey, and quail) usually show low bootstrap support values at short internal branches, reflecting the rapid diversification of these birds in the Eocene. However, given the key role of chicken and related poultry species in agricultural, evolutionary, general biological and disease studies, it is important to know their internal relationships. Recently, insertion patterns of transposable elements such as long and short interspersed nuclear element markers have proved powerful in revealing branching orders of difficult phylogenies. Here we decipher the order of speciation events in a group of 27 galliform species based on insertion events of chicken repeat 1 (CR1) transposable elements. Forty-four CR1 marker loci were identified from the draft sequence of the chicken genome, and from turkey BAC clone sequence, and the presence or absence of markers across species was investigated via electrophoretic size separation of amplification products and subsequent confirmation by DNA sequencing. Thirty markers proved possible to type with electrophoresis of which 20 were phylogenetically informative. The distribution of these repeat elements supported a single homoplasy-free cladogram, which confirmed that megapodes, cracids, New World quail, and guinea fowl form outgroups to Phasianidae and that quails, pheasants, and partridges are each polyphyletic groups. Importantly, we show that chicken is an outgroup to turkey and quail, an observation which does not have significant support from previous DNA sequence- and DNA-DNA hybridization-based trees and has important implications for evolutionary studies based on sequence or karyotype data from galliforms. We discuss the potential and limitations of using a genome-based retrotransposon approach in resolving problematic phylogenies among birds. PMID- 17077155 TI - Autologous haematopoeitic stem cell rescue (AHSCR) for severe rheumatic disease in children: guidance for BSPAR members--executive summary. PMID- 17077156 TI - Non-steroidal anti-inflammatory drugs--changes in prescribing may be warranted. PMID- 17077157 TI - Three months treatment of active spondyloarthritis with imatinib mesylate: an open-label pilot study with six patients. PMID- 17077158 TI - Arthritis as presenting manifestation of pure neuritic leprosy--a rheumatologist's dilemma. AB - OBJECTIVES: Leprosy classically presents with cutaneous and neurological manifestations. In diagnosed cases of leprosy, rheumatological involvement varies from 1% to 70%. A primary articular presentation without cutaneous manifestations is not yet known. Herein, we present our experience of five cases of leprosy that presented with predominant articular involvement in the absence of cutaneous manifestations. METHODS: The study was conducted in the Department of Clinical Immunology, Sanjay Gandhi Postgraduate Institute of Medical Sciences located in the state of Uttar Pradesh, one of the nine endemic states in India. Case records of patients with a definite diagnosis of leprosy were screened for the presenting manifestations, pattern of articular involvement, tenosynovitis, neurological signs and symptoms. Reports of nerve conduction study (NCS), nerve and synovial biopsy and other diagnostic tests were retrieved from laboratory records. Available radiographs were examined for evidence of juxta-articular osteopenia and erosions. RESULTS: Case records of 11,740 patients were screened, of which 28 had a diagnosis of leprosy. Twenty patients had presented with rheumatological complaints primarily. Five of the patients who presented with inflammatory arthritis with/without tenosynovitis (n = 4) and tenosynovitis alone (n = 1) had pure neuritic leprosy. All of these patients had thickened peripheral nerves and abnormal NCS. Sural nerve biopsy confirmed the diagnosis of leprosy in all these cases. CONCLUSION: A combination of tenosynovitis and thickened nerves in association with symmetric polyarthritis should raise a suspicion of leprosy even in the absence of cutaneous features. PMID- 17077159 TI - Better memory and neural efficiency in young apolipoprotein E epsilon4 carriers. AB - The apolipoprotein E (APOE) epsilon4 allele is the major genetic risk factor for Alzheimer's disease, but an APOE effect on memory performance and memory-related neurophysiology in young, healthy subjects is unknown. We found an association of APOE epsilon4 with better episodic memory compared with APOE epsilon2 and epsilon3 in 340 young, healthy persons. Neuroimaging was performed in a subset of 34 memory-matched individuals to study genetic effects on memory-related brain activity independently of differential performance. E4 carriers decreased brain activity over 3 learning runs, whereas epsilon2 and epsilon3 carriers increased activity. This smaller neural investment of epsilon4 carriers into learning reappeared during retrieval: epsilon4 carriers exhibited reduced retrieval related activity with equal retrieval performance. APOE isoforms had no differential effects on cognitive measures other than memory, brain volumes, and brain activity related to working memory. We suggest that APOE epsilon4 is associated with good episodic memory and an economic use of memory-related neural resources in young, healthy humans. PMID- 17077160 TI - Motivation modulates the activity of the human mirror-neuron system. AB - It is not known whether the mirror-neuron system is modulated by motivation, such as hunger. In this study, 2 groups of healthy participants underwent 2 functional magnetic resonance imaging scanning sessions separated by 1.5 h interval. During each session, participants were presented with video clips of another person grasping objects or grasping food. The first session was conducted after participants from group 1 had fasted. Then these participants were allowed to eat and were scanned again. Participants from group 2 had a meal before the first session. Food-related stimuli elicited specific hemodynamic response in the parahippocampal gyrus, orbitofrontal cortex, and amygdala, when participants were in a hungry state as compared with a satiated state. In addition, regions that belong to the mirror-neuron system, including the inferior frontal gyrus, and the posterior parietal cortex showed greater response when participants were hungry. Increased activity was also detected in the extrastriate body area. A positive correlation was observed between the self-report ratings of hunger and the hemodynamic activity in the inferior frontal gyrus as well as in the amygdala. Our results suggest that motivation to eat modulates the neural activity in the mirror-neuron system, facilitating the preparation or the intention to act. PMID- 17077162 TI - Proteinase 3 and CD177 are expressed on the plasma membrane of the same subset of neutrophils. AB - Proteinase 3 (PR3) is found in granules of all neutrophils but also on the plasma membrane of a subset of neutrophils (mPR3). CD177, another neutrophil protein, also displays a bimodal surface expression. In this study, we have investigated the coexpression of these two molecules, as well as the effect of cell activation on their surface expression. We can show that CD177 is expressed on the same subset of neutrophils as mPR3. Experiments show that the expression of mPR3 and CD177 on the plasma membrane is increased or decreased in parallel during cell stimulation or spontaneous apoptosis. Furthermore, we observed a rapid internalization and recirculation of mPR3 and plasma membrane CD177, where all mPR3 is replaced within 30 min. Our findings suggest that the PR3 found on the plasma membrane has its origin in the same intracellular storage as CD177, i.e., secondary granules and secretory vesicles and not primary granules. PR3- and CD177-expressing neutrophils constitute a subpopulation of neutrophils with an unknown role in the innate immune system, which may play an important role in diseases such as Wegener's granulomatosis and polycythemia vera. PMID- 17077163 TI - Ocular immune privilege is circumvented by CD4+ T cells, leading to the rejection of intraocular tumors in an IFN-{gamma}-dependent manner. AB - Although intraocular tumors reside in an immune-privileged site, they can circumvent immune privilege and undergo rejection, which typically follows one of two pathways. One pathway involves CD4(+) T cells, delayed-type hypersensitivity (DTH), and the culmination in ischemic necrosis of the tumor and phthisis (atrophy) of the eye. The second pathway is DTH-independent and does not inflict collateral injury to ocular tissues, and the eye is preserved. In this study, we used a well-characterized tumor, Ad5E1, to analyze the role of IFN-gamma in the nonphthisical form of intraocular tumor rejection. The results showed that IFN gamma induced tumor cell apoptosis, inhibited tumor cell proliferation, and promoted rejection by inhibiting angiogenesis. Microarray analysis revealed that IFN-gamma induced up-regulation of five antiangiogenic genes and down-regulation of four proangiogenic genes in Ad5E1 tumor cells. Although IFN-gamma knockout (KO) mice have progressively growing intraocular tumors, IFN-gamma was not needed for the elimination of extraocular tumors, as all IFN-gamma KO mice rejected s.c. tumor inocula. This represents a heretofore unrecognized role for IFN-gamma in circumventing ocular immune privilege and eliminating intraocular tumors. The findings also reveal that some IFN-gamma-independent tumor rejection processes are excluded from the eye and may represent a new facet of ocular immune privilege. PMID- 17077164 TI - Protein kinase C activity is required for cytotoxic T cell lytic granule exocytosis, but the theta isoform does not play a preferential role. AB - CTLs kill virus-infected, tumor, and transplanted targets via secretion of lytic agents including perforin and granzymes. Knowledge of the signals controlling this important process remains vague. We have tested the idea that protein kinase C (PKC)theta, a member of the novel PKC (nPKC) family, which has been shown to play a preferential role in critical Th cell functions, plays a similar, preferential role in CTL lytic granule exocytosis using T acute lymphoblastic leukemia-104 (TALL-104) human leukemic CTLs as a model. We provide evidence consistent with the idea that PKC activity is important for the degranulation step of lytic granule exocytosis, as opposed to upstream events. In contrast with previous work, our results with pharmacological agents suggest that conventional PKCs (cPKCs) and nPKCs may participate. Our results suggest that stimulation with soluble agents that bypass the TCR and trigger granule exocytosis activates PKCalpha and PKCtheta, which can both accumulate at the site of contact with a target cell, although PKCtheta did so more often. Finally, using a novel assay that detects granule exocytosis specifically in transfected, viable cells, we find that overexpression of constitutively active mutants of PKCalpha or PKCtheta can synergize with increases in intracellular [Ca(2+)] to promote granule exocytosis. Taken together, our results lend support for the idea that PKCtheta does not play a preferential role in CTL granule exocytosis. PMID- 17077161 TI - Attentional modulation of receptive field structure in area 7a of the behaving monkey. AB - Spatial attention modulates the activity of inferior parietal neurons. A statistically rigorous approach to classical retinotopic mapping was used to quantify the receptive fields of area 7a neurons under 2 attentional conditions. Measurements were made with retinal stimulation held constant and the locus of attention manipulated covertly. Both tasks required central fixation but differed in the locus of covert attention (either on the center fixation point or on a peripheral square target in one of 25 locations). The neuron's identity over the recording session was confirmed using chaos theory to characterize unique temporal patterns. Sixty-six percent of the neurons changed prestimulus activity based on task state. Retinotopic mapping showed no evidence for foveal sparing. Attentional factors influenced visual responses for approximately 30% of the neurons. Two types of modulation were equally observed. One group of cells had a multiplicative scaling of response, with equal instances of enhancement and suppression. A second group of cells had a complex interaction of visual and attentional signals, such that spatial tuning was subject to a nonlinear modulation across the visual field based on attentional constraints. These 2 cell groups may have different roles in the shift of attention preceding motor behaviors and may underlie shifts in parietal retinotopic maps observed with intrinsic optical imaging. PMID- 17077165 TI - Wild-type p53 inhibits nuclear factor-kappaB-induced matrix metalloproteinase-9 promoter activation: implications for soft tissue sarcoma growth and metastasis. AB - Human soft tissue sarcoma (STS) is a highly lethal malignancy in which control of metastasis determines survival. Little is known about the molecular determinants of STS dissemination. Here, we show that human STS express high levels of matrix metalloproteinase-9 (MMP-9) and that MMP-9 expression levels correlate with sequence analysis-defined p53 mutational status. Reintroduction of wild-type p53 (wtp53) into mutant p53 STS cell lines decreased MMP-9 mRNA and protein levels, decreased zymography-assessed MMP-9 proteolytic activity, and decreased tumor cell invasiveness. Reintroduction of wtp53 into STS xenografts decreased tumor growth and MMP-9 protein expression. Luciferase reporter studies showed that reintroduction of wtp53 into mutant p53 STS cells decreased MMP-9 promoter activity. Deletion constructs of the MMP-9 promoter identified a region containing a p53-responsive element that lacked a p53 consensus binding site but did contain a nuclear factor-kappaB (NF-kappaB) site. Mutating this NF-kappaB binding site eliminated the wtp53-repressive effect. Electrophoretic mobility shift assays confirmed decreased NF-kappaB binding in STS cells in the presence of wtp53. Our findings suggest a role for MMP-9 in STS progression and expand the role of p53 in molecular control of STS growth and metastasis. Therapeutic interventions in human STS targeting MMP-9 activity directly or via reintroduction of wtp53 merit further investigation. PMID- 17077166 TI - Improving measurement methods for behavior change interventions: opportunities for innovation. PMID- 17077167 TI - Item response modeling: an evaluation of the children's fruit and vegetable self efficacy questionnaire. AB - Perceived self-efficacy (SE) for eating fruit and vegetables (FV) is a key variable mediating FV change in interventions. This study applies item response modeling (IRM) to a fruit, juice and vegetable self-efficacy questionnaire (FVSEQ) previously validated with classical test theory (CTT) procedures. The 24 item (five-point Likert scale) FVSEQ was administered to 1578 fourth graders from 26 Houston schools. The IRM partial credit model indicated the five-point response options were not fully utilized. The questionnaire exhibited acceptable (>0.70) reliability except at the extremes of the SE scale. Differential item functioning (DIF) analyses revealed no response bias due to gender. However, DIF was detected by ethnic groups in 10 items. IRM of this scale expanded what was known from CTT methods in three ways: (i) areas of the scale were identified that were not as reliable, (ii) limitations were found in the response format and (c) areas of the SE scale levels were not measured. The FVSEQ can be improved by including items at the extreme levels of difficulty. DIF analyses identified areas where IRM can be useful to improve the functioning of measures. PMID- 17077168 TI - Neighborhood environment and risk of ischemic stroke: the brain attack surveillance in Corpus Christi (BASIC) Project. AB - The authors explored whether neighborhood-level characteristics are associated with ischemic stroke and whether the association differs by ethnicity, age, and gender. Using data from the Brain Attack Surveillance in Corpus Christi Project (January 2000-June 2003), they identified cases of ischemic stroke (n = 1,247) from both hospital and out-of-hospital sources. Census tracts served as proxies for neighborhoods, and neighborhood socioeconomic status scores were constructed from census variables (higher scores represented less disadvantage). In Poisson regression analyses comparing the 90th percentile of neighborhood score with the 10th, the relative risk of stroke was 0.49 (95% confidence interval (CI): 0.41, 0.58). After adjustment for age, gender, and ethnicity, this association was attenuated (relative risk (RR) = 0.79, 95% CI: 0.63, 1.00). There was no ethnic difference in the association of score with stroke (p for interaction = 0.79). Significant effect modification was found for age (p for interaction < 0.001) and gender (p for interaction = 0.04), with increasing scores being protective against stroke in men and younger persons. Associations were attenuated after adjustment for education (men: RR = 0.77, 95% CI: 0.55, 1.07; persons aged <65 years: RR = 0.65, 95% CI: 0.41, 1.02). Neighborhood characteristics may influence stroke risk in certain gender and age groups. Mechanisms for these associations should be examined. PMID- 17077169 TI - Prescription medications: beliefs, experiences, behavior, and adherence of sheltered housing residents. AB - BACKGROUND: Data on medication utilization among residents of sheltered housing complexes (SHCs) are limited. OBJECTIVE: To evaluate the extent of nonadherence to prescribed medications among residents of SHCs and identify factors associated with nonadherence. METHODS: A 58 item, pre-piloted questionnaire with validated items for patient self-assessment of medication risk; adherence; disability; health beliefs, experiences, and behavior; and general items on demographics, health, and medication use was mailed to 1137 SHC residents in Aberdeen, Scotland. Predictors of nonadherence (Morisky score >0) were identified using logistic regression. RESULTS: Of the 695 (61.1%) respondents, 645 (mean +/- SD age, 78.2 +/- 7.8 y) reported using prescribed drugs. Nonadherence was reported by 176 of 627 (28.1%) residents. The predictors of nonadherence were: disagreeing with the statement, "I ensure I have enough medicines so that I don't run out" (OR 0.48; 95% CI 0.30 to 0.77), agreeing with the statements, "I make changes in the recommended management to suit my lifestyle" (OR 1.32; 95% CI 1.09 to 1.60) and "I get confused about my medicines" (OR 1.39; 95% CI 1.10 to 1.77), younger age (OR 0.96; 95% CI 0.94 to 0.99), not getting help from someone to use the medications correctly (OR 2.20; 95% CI 1.21 to 3.99), and agreeing with the statement, "I am concerned about the side effects from my medicines" (OR 1.31; 95% CI 1.05 to 1.63). CONCLUSIONS: Over one-quarter of SHC residents were found to be nonadherent to prescribed drugs. Risk factors for nonadherence include younger age, confusion about drugs, lack of support for drug supply and administration, interference of treatment recommendations with lifestyle, and a perceived view of risks outweighing benefits in using recommended drug therapy. PMID- 17077170 TI - Ensuring that patients receive full cardiac pharmacotherapy services: a pharmacist's call to arms. AB - While clinical trials have proven that use of certain cardiovascular drugs improves morbidity and mortality, these drugs are not widely used, they are by and large prescribed in suboptimal doses, and their benefit is further diminished by nonadherence. Pharmacists have the ability to improve the survival of patients with cardiovascular risks. This can only be accomplished through the efforts of the individual pharmacist and the further development of health-system management services. Pharmacists need to leverage their access to patients, their large patient base, and their knowledge of refill dates to improve patient care. Support and development of more pharmacist-directed anticoagulation, hyperlipidemia, and hypertension clinics are also needed. PMID- 17077173 TI - Acid suppressive therapy and the effects on protease inhibitors. AB - OBJECTIVE: To review the literature associated with the pharmacokinetic interaction between protease inhibitors (PIs) and acid suppressive therapies and to characterize the impact of this interaction on virologic and immunologic outcomes. DATA SOURCES: A MEDLINE search (1966-October 2006) was conducted using the names of the 10 PIs and specific acid suppressive therapies including antacids, histamine(2)-receptor antagonists, and proton pump inhibitors. Abstracts and poster presentations from recent HIV/AIDS meetings were reviewed for relevance. References from retrieved articles, as well as product packaging and manufacturer information, were evaluated. STUDY SELECTION AND DATA EXTRACTION: Pertinent pharmacokinetic, immunologic, and virologic studies, in healthy and HIV-infected patients, evaluating the use of a PI and acid suppressive therapy were reviewed. DATA SYNTHESIS: Potential interactions between concomitant acid suppressive therapy and PIs were evaluated. Available information indicates that indinavir and atazanavir require an acidic gastric medium for adequate absorption. Indinavir pharmacokinetic parameters are variable with acid suppressive therapy but primarily result in decreased oral absorption. This interaction abates with concurrent ritonavir use. No immunologic or virologic data are available regarding the concomitant use of indinavir and acid suppressive therapy. The minimum concentration of atazanavir, area under the concentration-time curve, and maximum concentration are significantly reduced when used concurrently with acid suppressive therapy. Atazanavir 300 or 400 mg boosted with ritonavir 100 mg increases plasma concentrations when used with acid suppressive drugs. Virologic and immunologic outcomes appear stable when boosted atazanavir is used in HIV-positive patients. Atazanavir therapeutic monitoring should be considered when used in combination with acid suppressive therapy. CONCLUSIONS: Of the PIs reviewed, significant pharmacokinetic interactions exist between acid suppressive therapy and indinavir or atazanavir. These PIs should be used with low-dose ritonavir if acid suppressive therapy is necessary. PMID- 17077174 TI - Concomitant agranulocytosis and hepatotoxicity after treatment with carbimazole. AB - OBJECTIVE: To describe a case of agranulocytosis and severe hepatotoxicity associated with carbimazole treatment. CASE SUMMARY: A 37-year-old woman was diagnosed with severe hyperthyroidism resulting from Graves' disease. Treatment with carbimazole 30 mg/day was initiated. Within 15 days following the start of therapy, both minor (eg, pruritus, rash, urticaria, fever, arthralgias) and potentially life-threatening (eg, agranulocytosis, severe mixed hepatotoxicity with severe cholestatic jaundice) adverse effects developed. The patient's symptoms and laboratory abnormalities resolved following withdrawal of carbimazole. Treatment with other antithyroid drugs was not attempted, and (131)I ablation of the thyroid was successfully performed. Thyroid function was maintained with standard follow-up care. Agranulocytosis, identified following bone marrow biopsy, was treated with granulocyte colony-stimulating factor. DISCUSSION: Agranulocytosis and hepatotoxicity are rare adverse effects associated with carbimazole treatment and are usually dose- and age-related. The likelihood that carbimazole induced these undesirable events in our patient is rated as probable based on the Naranjo probability scale. We believe this case to be the first to describe minor and major adverse effects related to carbimazole therapy in a patient with Graves' disease. CONCLUSIONS: Major adverse effects associated with carbimazole are infrequent. However, clinicians need to be aware that the effects described here, including severe liver failure and bone marrow toxicity, may occur in patients receiving this drug. PMID- 17077176 TI - Injection of IL-12- and IL-18-encoding plasmids ameliorates the autoimmune pathology of MRL/Mp-Tnfrsf6lpr mice: synergistic effect on autoimmune symptoms. AB - IL-12 and IL-18 are mediators involved in the onset and progression of the autoimmune disease developing in MRL/Mp-Tnfrsf6(lpr) (lpr) mice, which display symptoms similar to the human systemic lupus erythematosus (SLE). The pathology is characterized by progressive lymphadenopathy and auto-antibody-mediated multiple organ failure, e.g. glomerulonephritis, or pneumonitis and a concomitant increase in serum levels for IFNgamma and tumor necrosis factor-alpha (TNFalpha). In this study, we intramuscularly injected lpr mice with plasmids encoding IL-12 and IL-18, either alone or in combination, in order to affect the development of the autoimmune disease. Five biweekly injections of the combined plasmids starting at 4-5 weeks of age diminished serum levels of TNFalpha and reduced the ability of lymphocytes from treated mice to produce IFNgamma in vitro. Injection of both plasmids synergistically attenuated the development of autoimmune syndromes, lymphoproliferation in secondary lymphoid organs, proteinuria and kidney damage, and pneumonitis. We conclude that IL-12 and IL-18 synergistically affect the pathogenesis of the T(h)1-dependent autoimmune syndrome of lpr mice and that approaches that target both IL-12 and IL-18 may be a therapeutic option in the treatment of autoimmune SLE. PMID- 17077175 TI - Lipid-containing mimetics of natural triggers of innate immunity as CTL-inducing influenza vaccines. AB - Anti-viral CD8(+) T cell responses can be induced using synthetic lipopeptides and a range of different lipid moieties have been examined in a variety of model systems and in man for this purpose. Nevertheless, only limited data exist on comparative efficacy of different lipopeptides in a single model of protection so that the optimal composition for vaccination purposes remains unknown. In this study, we examined different lipid structures from bacterial or non-bacterial sources coupled to peptides representing influenza viral epitopes recognized by CD8(+) and CD4(+) T cells. These were assessed in the context of intra-nasal (i.n.) immunization in the absence of added adjuvant. The strongest immunogens were those containing bacterially derived lipids that induced dendritic cell (DC) maturation via Toll-like receptor 2 (TLR2) binding. The number of DCs induced to mature in vitro was directly associated with the strength of the CD8(+) T cell mediated viral clearing responses in primed mice. Mice immunized with the TLR2 binding lipopeptides showed greatly enhanced numbers of specific IFN-gamma secreting CD8(+) T cells at the site of infection after i.n. exposure to virus, which resulted in enhanced protection of the pneumonic lung. Importantly, lipopeptide-pulsed DCs were able to induce the appropriate T cells, indicating that the self-adjuvanting effects could occur in the absence of free lipopeptide interacting with additional TLR2-bearing cells in vivo. This study defines a hierarchy of lipopeptide constructs that can program DC to prime memory CD8(+) T cells that on recall function to clear influenza virus from the infected lung. PMID- 17077177 TI - AILIM/ICOS-mediated elongation of activated T cells is regulated by both the PI3 kinase/Akt and Rho family cascade. AB - T-cell migration and movement is a critical component of a fully functional immune system. Activation-inducible lymphocyte immunomediatory molecule/inducible co-stimulator (AILIM/ICOS), which is a member of CD28 co-stimulatory receptor family, induces both activated T-cell migration underneath tumor necrosis factor alpha-treated human umbilical vein endothelial cell layers and also the morphological polarization of activated T cells. In our current study, we have investigated the signaling mechanisms underlying the morphological polarization of activated T cells, initiated by AILIM/ICOS signaling. AILIM/ICOS signaling induces the activation of phosphoinositide-3 (PI3)-kinase, the product of which, phosphatidylinositol 3,4,5-trisphosphate (PIP3), was found to be localized in the lamellipodia at the front part of the cells. Phosphorylated Akt is also co localized with PIP3 and filamentous actin in lamellipodia and the PI3-kinase/Akt signaling cascade has critical roles in T-cell polarization and lamellipodia formation via the re-organization of the actin cytoskeleton. Rho family members and their downstream effectors, Rho-associated kinase and p21-activated kinase (PAK), are also involved in AILIM/ICOS-mediated elongation. The PAK family members are serine/threonine kinase downstream effectors of both Rac and Cdc42. PAK3 is induced by the activation of T cells, whereas PAK1 is constitutively expressed in both naive and activated T cells. During the elongation, not only PAK1 but also PAK3 play an essential role through the phosphorylation of their conservative autophosphorylation sites and catalytic domain. Ser-244 phosphorylation, which is a putative Akt phosphorylation site, on PAK3 but not on PAK1 also regulates the morphological polarization of activated T cells by AILIM/ICOS signaling. Both the PI3-kinase/Akt and Rho family cascades operate coordinately to induce the forward migration of activated T cells by AILIM/ICOS signaling. PMID- 17077178 TI - Altered expression of vasoactive intestinal peptide receptors in T lymphocytes and aberrant Th1 immunity in multiple sclerosis. AB - Vasoactive intestinal peptide (VIP) has a unique property of regulating T(h)1 and T(h)2 immunity of CD4+ T cells. In this study, we demonstrated, for the first time, that differential expression of VIP receptors and a compensatory mechanism directly affect the responsiveness of CD4+ T cells and their T(h)1 and T(h)2 properties to VIP. The expression of VIP receptor-1 (VPAC1) and VPAC2 in CD4+ T cells changed reciprocally in the context of the activation state. In activated CD4+ T cells of healthy individuals, markedly decreased VPAC1 expression was compensated for by increased expression of VPAC2 induced by T cell activation. In contrast, there was altered expression of VPAC2 in activated CD4+ T cells derived from multiple sclerosis (MS) patients, which rendered CD4+ T cells less responsive to VIP and skewed the system to a predominantly in a T(h)1 direction. Detailed characterization with agonist peptides of VIP showed that residues Met and Ser at positions 17 and 25 of VIP were critical to its regulatory properties through interaction with VAPC2. Furthermore, altered levels of VPAC2 expression in T cells of MS patients were not associated with single-nucleotide polymorphism in the encoding region of the VPAC2 gene but with gene regulation as characterized by a distinct DNA footprinting pattern in the promoter region of the VPAC2 gene in MS as compared with controls. This study has provided new evidence for an intrinsic mechanism associated with an aberrant, pro-inflammatory state of CD4+ T cells in MS. PMID- 17077179 TI - Immunogenic HLA-B7-restricted peptides of hTRT. AB - Telomerase reverse transcriptase (TRT) is the first bona fide common tumor antigen. While several 9mer peptides of the human TRT have been identified for HLA-A2, little information exists on peptides for the remaining HLA types. Here, we used a multi-step approach to select and characterize a panel of HLA-B7 9mer peptides as candidate immunogens. In sequence, we used algorithm-based predictions, in vivo immunization of HLA-B7 transgenic (Tg) mice, in vitro immunization of human blood lymphocytes from two normal donors and two cancer patients, in vivo processing in HLA-B7 Tg mice and HLA-B7 supertype binding. We found a correlation between the in vivo immunogenicity and the actual HLA-B7 binding avidity of the seven predicted peptides. Furthermore, endogenous processing correlated with in vitro immunogenicity in human PBMC and HLA-B7 supertype binding. Peptide (1123)LPSDFKTIL(1131) (p1123) with the wider spectrum of supertype binding displayed the highest immunogenicity overall and was endogenously processed in several human lymphoblastoid cells. Since no single step of the screening/selection process could substitute for the whole approach, we conclude that the identification of MHC class I-restricted peptides for potential vaccination of cancer patients remains, by and large, an empirical process. PMID- 17077180 TI - Protein kinase D2 contributes to either IL-2 promoter regulation or induction of cell death upon TCR stimulation depending on its activity in Jurkat cells. AB - Members of protein kinase D (PKD) family serine/threonine kinases (PKD1, PKD2 and PKD3) are expressed in wide range of cells and regulate various cellular responses including immune responses. We have previously shown that PKD is involved in the signaling pathways of a human CD4(+) T cell clone stimulated with its cognate antigen. Contrary to foregoing publications, PKD1 mRNA was not detected in human T cells, Jurkat cells and mouse thymocytes and splenocytes. Instead, mass-spectrometric and reverse transcription-PCR analyses revealed that PKD2 was predominant in T cells. To investigate the roles of PKD2, wild-type (WT) and constitutively active (CA) PKD2 were expressed in Jurkat cells together with IL-2 promoter-driven reporter gene. Expression of WT-PKD2 enhanced IL-2 promoter activity upon stimulation with anti-CD3 mAb, while expression of CA-PKD2 inhibited IL-2 promoter activity and induced cell death. Although the cell death was suppressed by the treatment with caspase inhibitor, the IL-2 promoter activity was rarely recovered in CA-PKD2-expressing cells upon TCR stimulation. WT-PKD2 localized mainly in the cytoplasm translocated into the nucleus after TCR stimulation, while CA-PKD2 was present in both the cytoplasm and the nuclei before and after stimulation. Proteomic analyses revealed that CA-PKD2 enhanced the amount of phosphorylated SET protein, a histone chaperon that regulates histone acetylation, in Jurkat cells and the recombinant SET protein was phosphorylated by CA-PKD2 in vitro. The data provide a renewing insight into the subset of PKD family kinases expressed in T cells and suggest that PKD2 is involved in IL-2 promoter regulation and cell death depending on its activity upon TCR stimulation. PMID- 17077181 TI - Enhanced half-life of genetically engineered human IgG1 antibodies in a humanized FcRn mouse model: potential application in humorally mediated autoimmune disease. AB - The MHC class I-like Fc receptor FcRn plays an essential role in extending the half-life (t(1/2)) of IgG antibodies and IgG-Fc-based therapeutics in the circulation. The goal of this study was to analyze the effect of human IgG1 (hIgG1) antibodies with enhanced in vitro binding to FcRn on their in vivo t(1/2) in mice expressing human FcRn (hFcRn). Mutants of the humanized monoclonal Herceptin antibody (Hu4D5-IgG1), directed against human epidermal growth factor receptor 2 (p185 (HER2)), show altered pH-dependent binding to hFcRn in vitro. Two engineered IgG1 mutants (N434A and T307A/E380A/N434A) showed a considerably extended t(1/2) in vivo compared with wild-type antibody in mice expressing an hFcRn transgene (Tg) but not in mice expressing the endogenous mouse FcRn. The efficiency of hFcRn-mediated protection was dependent on hFcRn Tg copy number. Moreover, when injected into FcRn-humanized mice at a concentration sufficient to partially saturate hFcRn, the engineered IgG1 mutants with an extended serum t(1/2) were most effective in reducing the t(1/2) of a tracer hIgG1 antibody. Finally, administration of mutant with high binding to hFcRn ameliorated arthritis induced by passive transfer with human pathogenic plasma. These results indicate that Fc regions modified for high binding affinity to hFcRn increases serum persistence of therapeutic antibodies, that the same approach can be exploited as an anti-autoimmune therapy to promote the clearance of endogenous pathogenic IgG and that FcRn-humanized mice are a promising surrogate for hIgG therapeutic development. PMID- 17077182 TI - An early auxin-responsive Aux/IAA gene from wheat (Triticum aestivum) is induced by epibrassinolide and differentially regulated by light and calcium. AB - The plant hormone auxin plays a central role in regulating many aspects of plant growth and development. This largely occurs as a consequence of changes in gene expression. The Aux/IAA genes are best characterized among the early auxin responsive genes, which encode short-lived transcriptional repressors. In most plants examined, including Arabidopsis, soybean, and rice, the Aux/IAA genes constitute a large gene family. By screening the available databases, at least 15 expressed sequence tags (ESTs) have been identified from wheat (Triticum aestivum), which exhibit high sequence identity with Aux/IAA homologues in other species. One of these Aux/IAA genes, TaIAA1, harbouring all the four conserved domains characteristic of the Aux/IAA proteins, has been characterized in detail. The expression of TaIAA1 is light-sensitive, tissue-specific, and is induced within 15-30 min of exogenous auxin application. Also, the TaIAA1 transcript levels increase in the presence of a divalent cation, Ca(2+), and this effect is reversed by the calcium-chelating agent, EGTA. The TaIAA1 gene qualifies as the primary response gene because an increase in its transcript levels by auxin is unaffected by cycloheximide. In addition to auxin, the TaIAA1 gene is also induced by brassinosteroid, providing evidence that interplay between hormones is crucial for the regulation of plant growth and development. PMID- 17077183 TI - Ethylene-regulation of fruit softening and softening-related genes in peach. AB - To investigate the role of ethylene in peach fruit softening during ripening, stony hard peach fruit, in which ethylene production is suppressed during ripening, were treated with various concentrations of ethylene. There was no noticeable decrease in flesh firmness without ethylene treatment, while applied ethylene, in the range 0.1-100 microl l(-1), resulted in fruit softening. Furthermore, the fruit softened more rapidly when the applied ethylene concentration was higher. When ethylene treatment was interrupted, the degree of softening was greatly reduced. These results indicated that continuous ethylene treatment was required for the initiation and progression of fruit softening and that ethylene concentration is also an important factor in regulating the rate of softening. Eight genes, which putatively encode cell wall metabolism-related proteins, were investigated for mRNA accumulation patterns in the two different softening phenotypes of melting and stony hard peaches. All of the mRNAs investigated accumulated in fruit of the melting-flesh "Akatsuki" during ripening. By contrast, in the stony hard-flesh "Manami", the mRNAs for a putative endopolygalacturonase (PpPG), an alpha-L-arabinofuranosidase/beta-xylosidase (PpARF/XYL), and an expansin (PpExp3) showed either much lower levels or did not accumulate, and were identified as softening-related genes. Interruption of ethylene treatment indicated that these genes were regulated at the transcriptional level, and quickly responded to the presence or absence of ethylene before the softening response occurred, suggesting that ethylene directly regulates the transcription of these softening-related genes. These results suggested that cell wall metabolism, causing a rapid loss of firmness in peach fruit, may be controlled by ethylene at the transcriptional level. PMID- 17077184 TI - Cytokinins inhibit epiphyllous plantlet development on leaves of Bryophyllum (Kalanchoe) marnierianum. AB - When leaves of Bryophyllum marnierianum are detached from the plant, plantlets develop from primordia located at their margins. Leaves excised with a piece of stem attached do not produce plantlets. Severing the major leaf veins overcomes the inhibitory effect of the attached stem, indicating that the control agent is transmitted through the vascular system. A possible mechanism is that an inhibitory substance, possibly a known plant hormone, transported from the stem to the leaf, suppresses plantlet development. A number of hormones were tested for their ability to inhibit plantlet primordium development in whole isolated leaves. Auxins had no effect, indicating that apical dominance is not involved. The cytokinins zeatin, kinetin, and benzylaminopurine (BAP) strongly inhibited plantlet development, suggesting that they may be the or a factor involved in maintenance of plantlet primordium dormancy when the leaf is attached to the plant. This hypothesis was strongly supported by the finding that treatment of leaves attached to stems with a cytokinin antagonist (purine riboside) released the primordia from inhibition. In contrast to whole leaves, plantlet primordium development on leaf explants incubated on Murashige Skoog medium containing 3% sucrose was strongly stimulated by cytokinins. A possible explanation of these observations is that in whole leaves the cytokinin signal is transduced into an inhibitory signal whereas in the isolated primordium cytokinin has a direct stimulatory effect. The inhibitory cytokinin pathway must be dominant as long as the leaf is attached to the plant. A model is proposed which could explain these findings. This study points to a novel role of cytokinins in the maintenance of foliar plantlet primordium dormancy. PMID- 17077185 TI - Evidence that oxymorphone-induced increases in micronuclei occur secondary to hyperthermia. AB - Oxymorphone is a potent opioid analgesic. Oral administration of oxymorphone to rats at doses >or= 20 mg/kg and mice at 500 mg/kg produced an increase in micronucleated polychromatic erythrocytes (MPCEs). Oxymorphone does not produce chromosome aberrations in vitro, suggesting that the increased MPCEs in vivo may involve indirect mechanisms. Opioids are known to affect thermoregulatory mechanisms. Changes in body temperature can increase the incidence of MPCEs in rodents. Studies were conducted to examine the relationship between increased MPCEs in rats given oxymorphone and changes in body temperature. Single oral doses of oxymorphone associated with increased MPCEs (20 and 40 mg/kg) also produced a marked, rapid increase in body temperature. When animals were pretreated with sodium salicylate, peak body temperature was lower and returned to baseline more quickly than when oxymorphone was given alone. MPCEs were evaluated in rats after administration of oxymorphone (40 mg/kg) alone or following pretreatment with an oral dose of sodium salicylate. Oxymorphone alone produced a statistically significant increase in the incidence of MPCEs (3.6 per 1000 polychromatic erythrocytes vs. 0.4 in controls). The number of MPCEs in animals pretreated with sodium salicylate was similar to controls. Sodium salicylate alone had no effect on the number of MPCEs. Systemic oxymorphone exposure was not affected by sodium salicylate pretreatment; maximum plasma concentration (C(max)) and area-under-the-curve values were similar after administration of oxymorphone alone or following pretreatment with sodium salicylate. These results indicate that the increased incidence of MPCEs following oxymorphone administration is directly related to increased body temperature. PMID- 17077186 TI - Chronic exposure to a trichloroethylene metabolite in autoimmune-prone MRL+/+ mice promotes immune modulation and alopecia. AB - The industrial solvent trichloroethylene (TCE) is a widespread environmental contaminant known to impact the immune system. In the present study, female MRL+/+ mice were treated for 40 weeks with trichloroacetaldehyde hydrate (TCAH), a metabolite of TCE, in the drinking water. The results were compared with the data from an earlier study in which MRL+/+ mice were exposed to TCAH for 4 weeks. Following a 40-week exposure, the mice developed skin inflammation and dose dependent alopecia. In addition, TCAH appeared to modulate the CD4(+) T-cell subset by promoting the expression of an activated/effector (i.e., CD62L(lo)) phenotype with an increased capacity to secrete the proinflammatory cytokine interferon-gamma. However, unlike what was observed after only 4 weeks of exposure, TCAH did not significantly attenuate activation-induced cell death (AICD) or the expression of the death receptor FasL in CD4(+) T cells. Some metalloproteinases (MMPs) are thought to play a role in susceptibility to AICD by inducing FasL shedding. Thus, both the 4- and 40-week sera were tested for MMP-7 levels in an attempt to explain the disparate results of TCAH on AICD and FasL expression. Serum MMP-7 levels were significantly higher in mice exposed to TCAH for 4 weeks. In contrast, the serum MMP-7 levels were increased in all the mice by 40 weeks when compared with a nonautoimmune strain. Taken together, a chronic exposure to TCAH promotes alopecia and skin inflammation. The early effects of TCAH on MMP-7 levels may provide a mechanism by which TCAH promotes skin pathology. PMID- 17077187 TI - Phytochemicals induce breast cancer resistance protein in Caco-2 cells and enhance the transport of benzo[a]pyrene-3-sulfate. AB - We have previously reported that breast cancer resistance protein (BCRP) is involved in the transport of phase II metabolites of the food carcinogen benzo[a]pyrene (BP) in the human intestinal cell line Caco-2. Furthermore, the expression of BCRP seemed most likely to be aryl hydrocarbon receptor (AhR) dependent. Since numerous plant-derived anticarcinogens with AhR-agonistic activity have been identified to date, in the present study we investigated the effects of naturally occurring dietary compounds and tert-butyl hydroquinone (TBHQ) for their effects on BCRP expression. In Caco-2 cells, the most pronounced induction of BCRP expression could be observed after treatment with TBHQ (100 microM), dibenzoylmethane (DBM, 50 microM), and quercetin (25 microM), while green tea component (-)-epicatechin (50 microM) decreased BCRP expression. On mRNA level, quercetin, chrysin, flavone, and indole-3-carbinol showed a strong inducing effect, while genistein had no effect on BCRP mRNA expression. Curcumin and resveratrol showed a strong effect on BCRP induction in MCF-7 wild-type cells but no response in AhR-deficient MCF-7AHR(200) cells, supporting our hypothesis that BCRP is regulated via AhR-dependent signaling pathways. Inhibition of proteasome-mediated degradation of ligand-activated AhR caused a "superinduction" of BCRP mRNA. Antioxidant responsive element activators sulforaphane and diethylmaleate (DEM) had no inducing effect on BCRP mRNA expression. Caco-2 cells pretreated with quercetin or DBM showed an enhancement of apically transported benzo[a]pyrene-3-sulfate, indicating that induced BCRP was functionally active. In conclusion, apart from the modulation of detoxifying enzymes in the intestine, induction of BCRP by dietary constituents may contribute to the detoxification of food-derived procarcinogens such as BP. PMID- 17077188 TI - Fetal onset of aberrant gene expression relevant to pulmonary carcinogenesis in lung adenocarcinoma development induced by in utero arsenic exposure. AB - Arsenic is a human pulmonary carcinogen. Our work indicates that in utero arsenic exposure in mice can induce or initiate lung cancer in female offspring. To define early molecular changes, pregnant C3H mice were given 85 ppm arsenic in drinking water from days 8 to 18 of gestation and expression of selected genes in the fetal lung or in lung tumors developing in adults was examined. Transplacental arsenic exposure increased estrogen receptor-alpha (ER-alpha) transcript and protein levels in the female fetal lung. An overexpression of various estrogen-regulated genes also occurred, including trefoil factor-3, anterior gradient-2, and the steroid metabolism genes 17-beta-hydroxysteroid dehydrogenase type 5 and aromatase. The insulin growth factor system, which can be influenced by ER and has been implicated in the pulmonary oncogenic process, was activated in fetal lung after gestational arsenic exposure. In utero arsenic exposure also induced overexpression of alpha-fetoprotein, epidermal growth factor receptor, L-myc, and metallothionein-1 in fetal lung, all of which are associated with lung cancer. Lung adenoma and adenocarcinoma from adult female mice exposed to arsenic in utero showed widespread, intense nuclear ER-alpha expression. In contrast, normal adult lung and diethylnitrosamine-induced lung adenocarcinoma showed little evidence of ER-alpha expression. Thus, transplacental arsenic exposure at a carcinogenic dose produced aberrant estrogen linked pulmonary gene expression. ER-alpha activation was specifically associated with arsenic-induced lung adenocarcinoma and adenoma but not with nitrosamine induced lung tumors. These data provide evidence that arsenic-induced aberrant ER signaling could disrupt early life stage genetic programing in the lung leading eventually to lung tumor formation much later in adulthood. PMID- 17077189 TI - Studies on the physiological functions of the melanocortin system. AB - The melanocortin system refers to a set of hormonal, neuropeptidergic, and paracrine signaling pathways that are defined by components that include the five G protein-coupled melanocortin receptors; peptide agonists derived from the proopiomelanocortin preprohormone precursor; and the endogenous antagonists, agouti and agouti-related protein. This signaling system regulates a remarkably diverse array of physiological functions including pigmentation, adrenocortical steroidogenesis, energy homeostasis, natriuresis, erectile responses, energy homeostasis, and exocrine gland secretion. There are many complex and unique aspects of melanocortin signaling, such as the existence of endogenous antagonists, the agouti proteins, that act at three of the five melanocortin receptors. However, there is an aspect of melanocortin signaling that has facilitated highly reductionist approaches aimed at understanding the physiological functions of each receptor and peptide: in contrast to many peptides, the melanocortin agonists and antagonists are expressed in a limited number of very discrete locations. Similarly, the melanocortin receptors are also expressed in a limited number of discrete locations where they tend to be involved in rather circumscribed physiological functions. This review examines my laboratory's participation in the cloning of the melanocortin receptors and characterization of their physiological roles. PMID- 17077190 TI - Gut peptides in the regulation of food intake and energy homeostasis. AB - Gut hormones signal to the central nervous system to influence energy homeostasis. Evidence supports the existence of a system in the gut that senses the presence of food in the gastrointestinal tract and signals to the brain via neural and endocrine mechanisms to regulate short-term appetite and satiety. Recent evidence has shown that specific gut hormones administered at physiological or pathophysiological concentrations can influence appetite in rodents and humans. Gut hormones therefore have an important physiological role in postprandial satiety, and gut hormone signaling systems represent important pharmaceutical targets for potential antiobesity therapies. Our laboratory investigates the role of gut hormones in energy homeostasis and has a particular interest in this field of translational research. In this review we describe our initial studies and the results of more recent investigations into the effects of the gastric hormone ghrelin and the intestinal hormones peptide YY, pancreatic polypeptide, glucagon-like peptide-1, and oxyntomodulin on energy homeostasis. We also speculate on the role of gut hormones in the future treatment of obesity. PMID- 17077191 TI - The molecular control of corpus luteum formation, function, and regression. AB - The corpus luteum (CL) is one of the few endocrine glands that forms from the remains of another organ and whose function and survival are limited in scope and time. The CL is the site of rapid remodeling, growth, differentiation, and death of cells originating from granulosa, theca, capillaries, and fibroblasts. The apparent raison d'etre of the CL is the production of progesterone, and all the structural and functional features of this gland are geared toward this end. Because of its unique importance for successful pregnancies, the mammals have evolved a complex series of checks and balances that maintains progesterone at appropriate levels throughout gestation. The formation, maintenance, regression, and steroidogenesis of the CL are among the most significant and closely regulated events in mammalian reproduction. During pregnancy, the fate of the CL depends on the interplay of ovarian, pituitary, and placental regulators. At the end of its life span, the CL undergoes a process of regression leading to its disappearance from the ovary and allowing the initiation of a new cycle. The generation of transgenic, knockout and knockin mice and the development of innovative technologies have revealed a novel role of several molecules in the reprogramming of granulosa cells into luteal cells and in the hormonal and molecular control of the function and demise of the CL. The current review highlights our knowledge on these key molecular events in rodents. PMID- 17077192 TI - Progressive disorganization of the excitation-contraction coupling apparatus in aging human skeletal muscle as revealed by electron microscopy: a possible role in the decline of muscle performance. AB - An impairment of the mechanisms controlling the release of calcium from internal stores (excitation-contraction [EC] coupling) has been proposed to contribute to the age-related decline of muscle performance that accompanies aging (EC uncoupling theory). EC coupling in muscle fibers occurs at the junctions between sarcoplasmic reticulum and transverse tubules, in structures called calcium release units (CRUs). We studied the frequency, cellular localization, and ultrastructure of CRUs in human muscle biopsies from male and female participants with ages ranging from 28 to 83 years. Our results show significant alterations in the CRUs' morphology and cellular disposition, and a significant decrease in their frequency between control and aged samples: 24.4/100 microm(2) (n = 2) versus 11.6/100 microm(2) (n = 7). These data indicate that in aging humans the EC coupling apparatus undergoes a partial disarrangement and a spatial reorganization that could interfere with an efficient delivery of Ca(2+) ions to the contractile proteins. PMID- 17077193 TI - Oxidation-resistant membrane phospholipids can explain longevity differences among the longest-living rodents and similarly-sized mice. AB - Underlying causes of species differences in maximum life span (MLS) are unknown, although differential vulnerability of membrane phospholipids to peroxidation is implicated. Membrane composition and longevity correlate with body size; membranes of longer-living, larger mammals have less polyunsaturated fatty acid (PUFA). We determined membrane phospholipid composition of naked mole-rats (MLS > 28.3 years) and similar-sized mice (MLS = 3-4 years) by gas-liquid chromatography to assess if the approximately 9x MLS difference could be explained. Mole-rat membrane composition was unchanged with age. Both species had similar amounts of membrane total unsaturated fatty acids; however, mice had 9 times more docosahexaenoic acid (DHA). Because this n-3PUFA is most susceptible to lipid peroxidation, mole-rat membranes are substantially more resistant to oxidative stress than are mice membranes. Naked mole-rat peroxidation indices, calculated from muscle and liver mitochondrial membranes, concur with those predicted by MLS rather than by body size, suggesting that membrane phospholipid composition is an important determinant of longevity. PMID- 17077194 TI - Genetic modulation of hormone levels and life span in hybrids between laboratory and wild-derived mice. AB - Previously we showed that mouse stocks derived from wild-caught progenitors are long-lived relative to genetically heterogeneous mice derived from laboratory adapted strains. Here we replicate this life-span effect, and show that F2 hybrids between wild-derived and laboratory-derived stocks have intermediate survival patterns. Moreover, wild-derived mice are small, lean, and slow to mature, and have low serum insulin-like growth factor-I (IGF-I) relative to genetically heterogeneous mice. These traits, too, were at intermediate levels in the F2 hybrids. Furthermore, serum IGF-I at 6 months was a significant predictor of life span in two different populations of F2 hybrid mice. Pooling across stocks, life span was negatively correlated with body weight and serum IGF-I levels, and positively correlated with age at vaginal patency and serum leptin levels. Overall, these finding suggest that wild-derived mice harbor alleles that increase longevity, perhaps through effects on growth, maturation, and early-life hormone levels. PMID- 17077195 TI - Age-related influence on thiol, disulfide, and protein-mixed disulfide levels in human plasma. AB - In this study, plasma levels of both low-molecular-mass sulfhydryls/disulfides and mixed disulfides with proteins in 41 healthy humans aged 21-92 years were measured, with the aim of assessing whether there is a shift of the thiol/disulfide balance during aging and verifying some of the possible effects of the thiol imbalance. Our data suggest that aging is strictly correlated to a decrease in plasma glutathione and cysteinylglycine with the concomitant increase of most oxidized forms of thiols and a parallel increase in total cysteine and total homocysteine, probably due to an augmented efflux of these amino acids from various organs. The occurrence of two distinct regulatory systems for plasmatic pools of glutathione/cysteinylglycine on the one hand and cysteine/homocysteine on the other hand is hypothesized. PMID- 17077196 TI - Effect of aging on bone marrow-derived murine CD11c+CD4-CD8alpha- dendritic cell function. AB - Dendritic cells (DCs) are actively used as cellular adjuvant in cancer immunotherapy. However, although DC immunotherapies primarily target the elderly population, little is known about the effect of aging on DC functions. Here, we compared the T-cell stimulation, cytokine production, and tumor surveillance functions of bone marrow-derived CD11c(+)CD4(-)CD8alpha(-) DCs of old and young C57BL/6 mice. Old immature bone marrow-derived CD4(-)CD8alpha(-) DCs (imDCs) were 4 times less effective than were young DCs in stimulating syngeneic CD4(+) T-cell proliferation. Old imDCs also have decreased DC-specific/intracellular adhesion molecule type 3-grabbing, nonintegrin (DC-SIGN) expression compared to young DCs. Interestingly, mice treated with the ovalbumin peptide-pulsed young DCs exhibited significantly greater tumor regression than with ovalbumin peptide-pulsed old DCs. Old terminally differentiated bone marrow-derived DCs (tDC) also have increased interleukin-10, but decreased interleukin-6 and tumor necrosis factor alpha production. Taken together, these results have important implications in the clinical application of DC-based tumor immunotherapy in elderly persons. PMID- 17077197 TI - The five cardinal signs of inflammation: Calor, Dolor, Rubor, Tumor ... and Penuria (Apologies to Aulus Cornelius Celsus, De medicina, c. A.D. 25). PMID- 17077198 TI - Persistent changes in interleukin-6 and lower extremity function following hip fracture. AB - BACKGROUND: A hip fracture often heralds a period of functional decline in elderly persons. Although an inflammatory reaction would be expected following a hip fracture, whether the degree of this response is associated with adverse functional outcomes is unknown. METHODS: In a cohort from the Baltimore Hip Studies, women aged 65 years or older with a hip fracture were evaluated at 3 or 10 days (baseline) and 2, 6, and 12 months (follow-up) postfracture. Serum was analyzed for interleukin-6 (IL-6) level. A score of timed performance of 9 tasks, the Lower Extremity Gain Scale (LEGS) was calculated at each evaluation. We divided participants into tertiles based on their cytokine levels at 2, 6, and 12 months, and examined the LEGS score trajectories as a function of IL-6 tertile using generalized estimating equations, adjusting for age, prefracture function, body fat, pain, cognitive function, type of surgical repair, the number of in hospital complications, and the number of comorbid medical conditions. RESULTS: At baseline, 2, 6, and 12 months, respectively, 149, 95, 101, and 82 participants provided serum samples; of these participants 65, 78, and 59 also provided a LEGS measure at 2, 6, and 12 months, respectively. At 12 months postfracture the median (interquartile range) of serum IL-6 levels was 7.4 (4.0, 15.9) pg/mL. Participants in the lowest tertile of IL-6 level performed better on the LEGS than did those in the highest tertile (p =.008). At 12 months postfracture, participants in the lowest tertile scored 5.3 points better (95% confidence interval, 2.0-8.6) on the LEGS than did those in the highest tertile (p =.002). CONCLUSIONS: Higher IL-6 levels are adversely associated with recovery of lower extremity function after hip fracture. Factors that predict cytokine response and the potential mechanisms by which this effect is mediated warrant further study. PMID- 17077199 TI - The loss of skeletal muscle strength, mass, and quality in older adults: the health, aging and body composition study. AB - BACKGROUND: The loss of muscle mass is considered to be a major determinant of strength loss in aging. However, large-scale longitudinal studies examining the association between the loss of mass and strength in older adults are lacking. METHODS: Three-year changes in muscle mass and strength were determined in 1880 older adults in the Health, Aging and Body Composition Study. Knee extensor strength was measured by isokinetic dynamometry. Whole body and appendicular lean and fat mass were assessed by dual-energy x-ray absorptiometry and computed tomography. RESULTS: Both men and women lost strength, with men losing almost twice as much strength as women. Blacks lost about 28% more strength than did whites. Annualized rates of leg strength decline (3.4% in white men, 4.1% in black men, 2.6% in white women, and 3.0% in black women) were about three times greater than the rates of loss of leg lean mass ( approximately 1% per year). The loss of lean mass, as well as higher baseline strength, lower baseline leg lean mass, and older age, was independently associated with strength decline in both men and women. However, gain of lean mass was not accompanied by strength maintenance or gain (ss coefficients; men, -0.48 +/- 4.61, p =.92, women, -1.68 +/- 3.57, p =.64). CONCLUSIONS: Although the loss of muscle mass is associated with the decline in strength in older adults, this strength decline is much more rapid than the concomitant loss of muscle mass, suggesting a decline in muscle quality. Moreover, maintaining or gaining muscle mass does not prevent aging associated declines in muscle strength. PMID- 17077200 TI - Analysis of functional polymorphisms of metalloproteinase genes in persons with vascular dementia and Alzheimer's disease. AB - BACKGROUND: Vascular dementia (VAD) and Alzheimer's disease (AD) may share common neuropathological mechanisms. Matrix metalloproteinases (MMPs) may induce destruction of the extracellular matrix, neuronal dysfunction, and death. Increased expression of these molecules has been found in a number of neurological diseases, including cerebral ischemia and AD. Expression and activity of MMPs may be genetically influenced by common polymorphisms in the promoter regions of the corresponding genes. The purpose of this study was to evaluate whether functional polymorphisms of MMP genes are associated with dementia. METHODS: This is a cross-sectional study including a total of 599 individuals: 193 with VAD, 183 with AD, and 223 controls. Polymorphisms of the MMP-1, MMP-3, and MMP-9 genes were studied. RESULTS: MMP-1 2G2G, MMP-1 1G2G, MMP 3 5A5A, and MMP-9 TT genotypes were significantly and independently associated with VAD (odds ratio [OR] 2.5, 95% confidence interval [CI] 1.4-4.4, OR = 1.7, 95% CI, 1.0-2.7, OR = 2.9, 95% CI, 1.5-5.9, and OR = 6.8, 95% CI, 1.3-35.1, respectively). MMP-1 2G2G and MMP-3 5A5A genotypes were associated with increased risk of AD only in persons who carry the apolipoprotein E (APOE) epsilon4 allele (OR = 6.0, 95% CI, 2.3-15.5, and OR = 14.3, 95% CI, 3.2-63.0, respectively). Interestingly, the odds of VAD and AD was further increased in persons concomitantly carrying more than one MMP gene variation, compared to individuals that only had one high-risk genotype. CONCLUSIONS: Our study suggests that MMP gene polymorphisms are associated with VAD and AD, although these results need to be treated with caution until replicated. MMP genotypes may influence the risk of dementia and merit further investigation as potential genetic markers of disease. PMID- 17077201 TI - Predicting mortality from clinical and nonclinical biomarkers. AB - BACKGROUND: Few studies focus on "preclinical" warning signs associated with mortality. In this article, we investigate associations between all-cause mortality and two clusters of biological risk factors: (i) standard clinical measures related to cardiovascular disease and metabolic function; and (ii) nonclinical measures pertaining to hypothalamic-pituitary-adrenal axis activity, sympathetic nervous system activity, and inflammatory response. METHODS: Data come from the 2000 Social Environment and Biomarkers of Aging Study, a national sample of Taiwanese persons aged 54 years or older; 1497 persons were interviewed in their homes, and 1023 participated in a hospital examination. The analysis is based on 927 respondents with complete information. Logistic regression models describe the association between biomarkers and the 3-year probability of dying. RESULTS: Although both groups of biomarkers are significantly associated with mortality, the model with neuroendocrine and immune biomarkers has better explanatory and discriminatory power than the one with clinical measures. The association between these nonclinical measures and mortality remains strong after adjustment for the clinical markers, suggesting that the physiological effects of the nonclinical biomarkers are broader than those captured by the cardiovascular and metabolic system measures included here. CONCLUSIONS: Nonclinical markers are likely to provide warning signs of deteriorating health and function beyond what can be learned from conventional markers. Our findings are consistent with those of recent studies that (i) demonstrate the importance of neuroendocrine and immune system markers for survival, and (ii) indicate that standard clinical variables are less predictive of mortality in older than in younger populations. PMID- 17077202 TI - The influence of age on the effects of lifestyle modification and metformin in prevention of diabetes. AB - BACKGROUND: The incidence of type 2 diabetes increases with age. It is unknown whether interventions to prevent diabetes are as effective in elderly persons as in younger adults. METHODS: The Diabetes Prevention Program (DPP) demonstrated that an intensive lifestyle intervention (ILS) or metformin could prevent or delay diabetes. A predefined secondary outcome of DPP was to determine if treatment effects varied by age. RESULTS: At baseline, participants aged 60-85 years were leaner and had the best insulin sensitivity and lowest insulin secretion compared to younger age groups. Diabetes incidence rates did not differ by age in the placebo group, but ILS was more effective with increasing age (6.3, 4.9, and 3.3 cases per 100 person-years, in the 25-44, 45-59, and 60-85 year age groups, respectively; p(trend) =.007). Participants aged 60-85 years had the most weight loss and metabolic equivalent (MET)-hours of physical activity. The metformin group showed a trend toward higher diabetes incidence among older participants (6.7, 7.7, and 9.3 cases per 100 person-years in the 25-44, 45-59, and 60-85 year age groups, respectively; p(trend) =.07); and diabetes risk increased with age (hazard ratio [age 60-85 vs 25-44] 1.63, p =.02), after adjusting for the greater weight loss in the 60-85 year age group. CONCLUSIONS: Lifestyle modification was exceptionally effective in preventing diabetes in older individuals; this finding was largely explained by greater weight loss and physical activity. The limited effectiveness of metformin in older persons may reflect age-related differences in insulin action and secretion. A lifestyle modification program can be recommended for older individuals at high risk for type 2 diabetes. PMID- 17077203 TI - A twin study on the heritability of walking ability among older women. AB - BACKGROUND: This study examined the role of genetic and environmental factors explaining individual differences in women's walking ability in old age. METHODS: A maximal walking speed test over 10 meters and a 6-minute walking endurance test were done under standard conditions among 92 monozygotic and 105 dizygotic pairs of twin sisters reared together, aged 63-75 years. RESULTS: The mean maximum walking speed was 1.73 +/- 0.32 m/s and the mean distance covered in the 6-minute walking test was 525.6 +/- 77.3 m. Multivariate genetic modeling showed that a minor part of the variances in walking speed (16%, 95% confidence interval [CI]: 0%-54%) and endurance (20%, 95% CI: 0%-56%) were accounted for by genetic influences, and that the genetic influences were common to both traits. The corresponding proportions for common environmental factors were 37% (95% CI: 4% 58%) and 26% (95% CI: 0%-52%), and for individual environmental factors 46% (95% CI: 35%-59%) and 54% (95% CI 42%-68%), respectively. The environmental effects were partially common to both traits. CONCLUSIONS: Among relatively healthy older women, a modest portion of the variances of walking speed and endurance were accounted for by genetic factors, whereas shared and individual environmental factors explained most of the variance in both traits. PMID- 17077204 TI - Glucose-6-phosphate dehydrogenase deficiency in female octogenarians, nanogenarians, and centenarians. AB - BACKGROUND: Age-related skewing of X-chromosome inactivation leading to glucose-6 phosphate dehydrogenase (G6PD) deficiency in elderly women in a population with prevalent G6PD gene mutations was investigated. METHODS: G6PD activity was measured biochemically. G6PD mutations were detected by polymerase chain reaction (PCR) and allele-specific extension, and analyzed by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry and Sequenom MassARRAY. X-chromosome inactivation was quantified by semiquantitative PCR for the HUMARA gene, before and after HpaII digestion. RESULTS: In 173 women (median age: 90 years; range, 80-107 years), 18 heterozygotes for G6PD mutations were identified. Three heterozygotes were G6PD deficient, owing to skewed X-chromosome inactivation affecting the wild-type allele. Fifteen heterozygotes, with skewing apparently affecting the mutant alleles, had normal but significantly lower G6PD levels. At 1.73%, G6PD deficiency was significantly more frequent than expected from population screening at birth. CONCLUSION: Due to skewed X-chromosome inactivation, elderly women in populations with prevalent G6PD mutations are at risk of G6PD deficiency. PMID- 17077205 TI - Falls risk and functional decline in older fallers discharged directly from emergency departments. AB - BACKGROUND: There is currently no standard approach to falls risk assessment and management for older fallers presenting to the emergency department (ED) who are discharged directly home. Hence, this study was conducted to describe the prevalence of falls risk factors associated with older fallers presenting to the ED and to identify the factors associated with postdischarge decline in function in this group. METHODS: This cross-sectional study was performed with 300 community-dwelling individuals, aged 60 years or older, admitted to the ED following a fall, and discharged directly home. A home-based assessment after ED discharge was performed, which included the prevalence of falls risk factors, identification of functional decline, and objective measurements of balance, gait, depression, and falls efficacy. RESULTS: Fall-related injuries were sustained by 91% (95% confidence interval [CI], 87.2%-94.0%) of participants presenting to the ED. The most common falls risk factors identified in the home assessment were polypharmacy (79.0%, 95% CI, 73.9%-83.5%), home hazards (76.0%, 95% CI, 70.8%-80.7%), decreased balance (61.3%, 95% CI, 55.6%-66.9%), and arthritis (61.3%, 95% CI, 55.6%-66.9%). A decline in function was reported by 35% of participants (95% CI, 29.6%-40.7%). Sustaining a fracture, functional independence before the fall, being female, depression, and slower Timed Up and Go (TUG) scores were associated with a decline in function (p <.05). CONCLUSION: Older fallers discharged directly from the ED have a high prevalence of falls risk factors and are at risk of functional decline. PMID- 17077206 TI - The ordering of milestones in language development for children from 1 to 6 years of age. AB - PURPOSE: To scale language milestones in a group of 527 children to provide an instrument for screening language development. Procedure The questionnaire regarding these milestones was completed by parental report. It was evaluated whether the scaled milestones satisfied the assumptions of the Mokken item response model. RESULTS: The scalability of the final scale of 14 milestones was strong (H = .95), its reliability was high (rho = .96), and it satisfied the assumptions of the Mokken model. CONCLUSIONS: A single, unidimensional scale of diverse milestones was developed. It taps lexical, syntactic, and phonological skills, as well as both receptive and expressive language skills, and is well suited for mapping progress in language ability. PMID- 17077207 TI - Factors that influence lexical and semantic fast mapping of young children with specific language impairment. AB - PURPOSE: This purpose of this study was to investigate the lexical and semantic fast mapping ability of young children with specific language impairment (SLI) and normal language (NL), with a specific emphasis on the influence of phonological factors. METHOD: The study included 46 children (mean age 58 months), half with SLI and half with NL. Children were asked to fast map visual information only, visual-plus-nonlinguistic-auditory information, and visual-plus linguistic-auditory information. A mixed design was used to compare children across and within groups. RESULTS: Children with SLI performed worse than children with NL overall. The SLI group showed specific deficits in semantic fast mapping when they saw visual information only. This condition may have disrupted encoding because it varied from the expected auditory and visual pattern. The children with SLI also performed poorly when they were asked to map phonotactically infrequent linguistic information and when the difficulty of the task increased. A nonword repetition task was correlated with both semantic and lexical fast mapping. CONCLUSIONS: The findings are discussed in the light of their support for a limited capacity model of processing, as well as the impact of phonology on word learning. PMID- 17077208 TI - The relationship between phonological memory, receptive vocabulary, and fast mapping in young children with specific language impairment. AB - PURPOSE: This study assessed the fast mapping performance of children with specific language impairment (SLI) across the preschool to kindergarten age span in relation to their phonological memory and vocabulary development. METHOD: Fifty-three children diagnosed with SLI and 53 children with normal language (NL) matched for age and gender (30 three-year-olds, 18 four-year-olds, 28 five-year olds, and 30 six-year-olds) participated. Children's phonological memory was assessed using nonword repetition and digit span tasks. Receptive vocabulary was assessed using the Peabody Picture Vocabulary Test-III. Children learned the names for 8 objects during 2 fast mapping tasks. RESULTS: Overall, the NL group demonstrated significantly better performance on phonological memory and vocabulary measures across the age span; however, performance on the fast mapping task differed significantly only at age 5. Phonological memory and existing receptive vocabulary did not predict fast mapping ability. CONCLUSIONS: The phonological memory skills of preschoolers with NL and SLI followed a similar developmental pattern, but the SLI group consistently scored significantly lower than the NL group. Overall, the NL group showed significantly better receptive vocabulary, with evidence that between-group differences increased at age 6. Neither short-term phonological memory nor receptive vocabulary predicted fast mapping comprehension or production performance, even though both have been shown to correlate with later stages of word learning. PMID- 17077209 TI - Nonword repetition: a comparison of tests. AB - PURPOSE: This study compared performance of children on 2 tests of nonword repetition to investigate the factors that may contribute to the well-documented nonword repetition deficit in specific language impairment (SLI). METHOD: Twelve children with SLI age 7 to 11 years, 12 age-matched control children, and 12 control children matched for language ability completed 2 tests of nonword repetition: the Children's Test of Nonword Repetition (CNRep) and the Nonword Repetition Test (NRT). RESULTS: The children with SLI performed significantly more poorly on both tests than typically developing children of the same age. The SLI group was impaired on the CNRep but not the NRT relative to younger children with similar language abilities when adjustments were made for differences in general cognitive ability. The children with SLI repeated the lengthiest nonwords and the nonwords containing consonant clusters significantly less accurately than the control groups. CONCLUSION: The evidence suggests that the nonword repetition deficit in SLI may arise from a number of factors, including verbal short-term memory, lexical knowledge, and output processes. PMID- 17077210 TI - Individual differences in the onset of tense marking: a growth-curve analysis. AB - The purpose of this study was to explore individual differences in children's tense onset growth trajectories and to determine whether any within- or between child predictors could account for these differences. Twenty-two children with expressive vocabulary abilities in the low-average to below-average range participated. Sixteen children were at risk for specific language impairment (SLI), and 6 children had low-average language abilities. Spontaneous language samples, obtained at 3-month intervals between 2;0 and 3;0, were analyzed to examine change in a cumulative productivity score for 5 tense morphemes: third person singular present, past tense, copula BE, auxiliary BE, and auxiliary DO. Hierarchical linear modeling was used to model intercept and linear growth at 30 months and quadratic growth overall. A growth model that included mean length of utterance (MLU) and MLU growth better explained within-child productivity score growth trajectories than a parallel model with vocabulary and vocabulary growth. Significant linear growth in productivity scores remained even after a control for MLU was in place. When between-child predictors were added in the final conditional model, only positive family history approached statistical significance, improving the overall estimation of the model's growth parameters. The findings support theoretical models of language acquisition that claim relative independence of tense marking from other more general aspects of vocabulary development and sentence length. The trends for family history are also consistent with proposals implicating faulty genetic mechanisms underlying developmental language disorders. Systematic use of familial risk data is recommended in future investigations examining the relationship between late talking children and children at risk for SLI. PMID- 17077211 TI - The effect of word frequency on noun and verb definitions: a developmental study. AB - PURPOSE: Word frequency has profound effects in word recognition and production tasks. Here the influence of word frequency on definitions was investigated, and it was hypothesized that word frequency would have significant influence on responses provided for definitions of nouns and verbs. METHOD: Students from Grades 4, 7, 10, and college wrote definitions for high- and low-frequency nouns and verbs and rated their familiarity with the stimulus words. In the noun study, definitions were coded with the semantic response categories "use/purpose," "description," "association/relation," "partial explanation," "explanation," and "error." "Partial explanation" and "explanation" responses were subcategorized to code for use of critical attributes of meaning and class terms. In the verb study, definitions were coded for "synonym," "association/relation," "class," "partial explanation," "explanation," and "error." RESULTS: Results indicated that certain response categories (such as a class term or a critical attribute) were more characteristic in definitions of high- compared with low-frequency words, whereas responses (such as nonspecific class) were more characteristic in low- compared with high-frequency words. In addition, certain response categories increased with age (such as use of class terms and critical attributes), while errors decreased with age. Familiarity ratings served to validate the high- and low- frequency nature of the stimulus words. CONCLUSIONS: In general, word frequency had a significant impact on word definitions. Implications are discussed with respect to word familiarity, representation in the mental lexicon, acquisition of word meaning, and shared linguistic knowledge. PMID- 17077212 TI - Teachers and laypersons discern quality differences between narratives produced by children with or without SLI. AB - PURPOSE: To examine the functional impact of specific language impairment (SLI). Specific goals were to determine whether (a) subjective ratings of narrative quality differentiate children with SLI from their normally developing (ND) age mates, (b) laypersons and teachers differ in their ratings of narrative quality, (c) objective measures confirm previously reported problems in narration among children with SLI, and (d) objective measures of narrative structure and quality ratings relate. METHOD: Twenty-seven laypersons and 21 teachers used interval scaling to rate the quality of narratives produced by 20 5-7-year-olds, 10 with SLI and 10 ND age-mates. The narratives were also analyzed objectively for fluency, length, sentence-level syntax, and story grammar and themes. RESULTS: Subjective ratings differentiated the SLI and ND groups with 70% nonoverlap. No differences were observed between the laypersons' and teachers' numeric ratings; however, laypersons reported that they paid more attention to the "sparkle" or charm of the narratives. Objective measures of story length, grammaticality, and thematic development differentiated SLI and ND groups. Mean length of C-unit and number of thematic units positively predicted quality ratings. Clinical implications Intervention efforts aimed specifically at improving the quality of these children's oral narration may focus on increasing length, grammatical accuracy, and story development. Future clinical and research efforts aimed at addressing the broader functional impact of SLI are also critical given that the manifestations of SLI are noticeable to both teachers and laypersons. PMID- 17077213 TI - Dynamic assessment of school-age children's narrative ability: an experimental investigation of classification accuracy. AB - Two experiments examined reliability and classification accuracy of a narration based dynamic assessment task. PURPOSE: The first experiment evaluated whether parallel results were obtained from stories created in response to 2 different wordless picture books. If so, the tasks and measures would be appropriate for assessing pretest and posttest change within a dynamic assessment format. The second experiment evaluated the extent to which children with language impairments performed differently than typically developing controls on dynamic assessment of narrative language. METHOD: In the first experiment, 58 1st- and 2nd-grade children told 2 stories about wordless picture books. Stories were rated on macrostructural and microstructural aspects of language form and content, and the ratings were subjected to reliability analyses. In the second experiment, 71 children participated in dynamic assessment. There were 3 phases: a pretest phase, in which children created a story that corresponded to 1 of the wordless picture books from Experiment 1; a teaching phase, in which children attended 2 short mediation sessions that focused on storytelling ability; and a posttest phase, in which children created a story that corresponded to a second wordless picture book from Experiment 1. Analyses compared the pretest and posttest stories that were told by 2 groups of children who received mediated learning (typical and language impaired groups) and a no-treatment control group of typically developing children from Experiment 1. RESULTS: The results of the first experiment indicated that the narrative measures applied to stories about 2 different wordless picture books had good internal consistency. In Experiment 2, typically developing children who received mediated learning demonstrated a greater amount of pretest to posttest change than children in the language impaired and control groups. Classification analysis indicated better specificity and sensitivity values for measures of response to intervention (modifiability) and posttest storytelling than for measures of pretest storytelling. Observation of modifiability was the single best indicator of language impairment. Posttest measures and modifiability together yielded no misclassifications. CONCLUSION: The first experiment supported the use of 2 wordless picture books as stimulus materials for collecting narratives before and after mediation within a dynamic assessment paradigm. The second experiment supported the use of dynamic assessment for accurately identifying language impairments in school-age children. PMID- 17077214 TI - Reliability issues and solutions for coding social communication performance in classroom settings. AB - PURPOSE: To explore the utility of time-interval analysis for documenting the reliability of coding social communication performance of children in classroom settings. Of particular interest was finding a method for determining whether independent observers could reliably judge both occurrence and duration of ongoing behavioral dimensions for describing social communication performance. METHOD: Four coders participated in this study. They observed and independently coded 6 social communication behavioral dimensions using handheld computers. The dimensions were mutually exclusive and accounted for all verbal and nonverbal productions during a specified time frame. The technology allowed for coding frequency and duration for each entered code. Data were collected from 20 different 2-min video segments of children in kindergarten through 3rd-grade classrooms. Data were analyzed for interobserver and intraobserver agreements using time-interval sorting and Cohen's kappa. Further, interval size and total observation length were manipulated to determine their influence on reliability. RESULTS: The data revealed interval sorting and kappa to be a suitable method for examining reliability of occurrence and duration of ongoing social communication behavioral dimensions. Nearly all comparisons yielded medium to large kappa values; interval size and length of observation minimally affected results. Implications The analysis procedure described in this research solves a challenge in reliability: comparing coding by independent observers of both occurrence and duration of behaviors. Results indicate the utility of a new coding taxonomy and technology for application in online observations of social communication in a classroom setting. PMID- 17077215 TI - Effect of attentional state on frequency discrimination: a comparison of children with ADHD on and off medication. AB - Debate continues over the hypothesis that children with language or literacy difficulties have a genuine auditory processing deficit. Several recent studies have reported deficits in frequency discrimination (FD), but it is unclear whether these are genuine perceptual impairments or reflective of the comorbid attentional problems that exist in many children with language and literacy difficulties. The present study investigated FD in children with attention deficit hyperactivity disorder (ADHD) when their attentional state was altered with stimulant medication. Auditory thresholds were obtained using FD and frequency modulation detection (FM) tasks. In the FD task, participants judged which of 2 pairs contained a high-low frequency sound, and in the FM task, children judged which of two tones "wobbled" (i.e., modulated). Children with ADHD had significantly poorer and more variable FD performance when off compared to on stimulant medication, and did significantly worse than controls on all FD runs when off but not on stimulant medication. However, children with ADHD did not differ from controls on the FM task. These findings demonstrate that certain auditory discrimination tasks are influenced by the child's attentional status. In addition, significant relationships between FD and measures of language and reading were abolished when comorbid attentional difficulties were taken into account. The study has implications for design and interpretation of studies investigating links between auditory discrimination and difficulties in language and literacy. PMID- 17077216 TI - Stability of low-frequency residual hearing in patients who are candidates for combined acoustic plus electric hearing. AB - The purpose of this study was to investigate the stability over time of low frequency auditory thresholds to better determine if the new technique of using a short-electrode cochlear implant that preserves residual low-frequency acoustic hearing can be a long-term solution for those with severe-to-profound hearing loss at high frequencies. The present study determined the long-term rate of decline in acoustic hearing in patients who have a preexisting hearing loss yet have not been implanted with a cochlear implant. A retrospective analysis of patients' audiograms that fit into the range for candidacy for the short electrode device was performed to calculate the rate of change of threshold over time. The analysis of adult patients' data indicated that there was an average of only 1.05 dB hearing deterioration per year in the low frequencies and that presbycusis accounted for approximately one third to one half of this decline. The average deterioration of hearing threshold for pediatric patients was 1.2 dB per year; however, the rates of change in pediatric patients were considerably more variable (across individuals and across frequencies) than in adults. These data provide support for the idea that the short-electrode cochlear implant may be a practical solution for most adults in the long run, but this may not be the case for all pediatric patients. PMID- 17077217 TI - Song recognition by children and adolescents with cochlear implants. AB - PURPOSE: To assess song recognition and pitch perception in prelingually deaf individuals with cochlear implants (CIs). METHOD: Fifteen hearing children (5-8 years) and 15 adults heard different versions of familiar popular songs-original (vocal + instrumental), original instrumental, and synthesized melody versions and identified the song in a closed-set task (Experiment 1). Ten CI users (8-18 years) and age-matched hearing listeners performed the same task (Experiment 2). Ten CI users (8-19 years) and 10 hearing 8-years-olds were required to detect pitch changes in repeating-tone contexts (Experiment 3). Finally, 8 CI users (6 19 years) and 13 hearing 5-year-olds were required to detect subtle pitch changes in a more challenging melodic context (Experiment 4). RESULTS: CI users performed more poorly than hearing listeners in all conditions. They succeeded in identifying the original and instrumental versions of familiar recorded songs, and they evaluated them favorably, but they could not identify the melody versions. Although CI users could detect a 0.5-semitone change in the simple context, they failed to detect a 1-semitone change in the more difficult melodic context. CONCLUSION: Current implant processors provide insufficient spectral detail for some aspects of music perception, but they do not preclude young implant users' enjoyment of music. PMID- 17077218 TI - The development of temporal resolution: between-channel gap detection in infants and adults. AB - PURPOSE: Infants have a good ability to detect brief silent gaps between 2 short identical sound markers (within-channel gap detection), with thresholds between 2 and 11 ms. The present experiment traces the development of temporal resolution for between-channel gaps (i.e., gaps delineated by spectrally disparate markers). This ability appears crucial for the perception of complex stimuli such as speech and is thought to reflect more central auditory processing. METHOD: Infants age 6 7.5 months and adults were tested in a between-channel gap detection task using a conditioned head-turn procedure. Gaps were marked by 1- and 4-kHz Gaussian enveloped sine-tone markers. RESULTS: Infant gap thresholds were between 30 and 40 ms under conditions in which adult thresholds were between 10 and 20 ms. CONCLUSIONS: Unlike within-channel gap detection, the central temporal processing required for between-channel gap detection is still immature at 6 months of age. PMID- 17077219 TI - Birth control pills and nonprofessional voice: acoustic analyses. AB - PURPOSE: Two studies are presented here. Study 1 was aimed at evaluating whether the voice characteristics of women who use birth control pills that contain different progestins differ from the voice characteristics of a control group. Study 2 presents a meta-analysis that combined the results of Study 1 with those from 3 recent studies that compared voices of women who use and do not use birth control pills. METHOD: In Study 1, voice samples from 30 women with no history of voice training, who use pills with different progestins (drospirenone, desogestrel, gestodene), and 10 women who do not use the pill were recorded at specific time points across the menstrual cycle and were analyzed acoustically. In Study 2, results from Study 1 were analyzed jointly with results from three recent studies, which used similar methodologies. RESULTS: Results of Study 1 did not reveal acoustic differences in sustained phonation of vowels across the pill groups and controls. Results of the meta-analysis performed in Study 2 indicated that pill users exhibited lower jitter and shimmer values on sustained vowels, whereas no difference of fundamental frequency was observed among women who use the pill. CONCLUSIONS: These results support findings from previous studies, which suggested that no adverse effect on voice was detected among nonprofessional speakers who use new-generation monophasic birth control pills, for the measures studied. Furthermore, results of the meta-analysis suggested that some acoustic properties of the voice, which are reflected in perturbation measures in sustained vowels, may be improved among women who use the pill. PMID- 17077220 TI - Alterations in CNS activity induced by botulinum toxin treatment in spasmodic dysphonia: an H215O PET study. AB - Speech-related changes in regional cerebral blood flow (rCBF) were measured using H(2)(15)O positron-emission tomography in 9 adults with adductor spasmodic dysphonia (ADSD) before and after botulinum toxin (BTX) injection and 10 age- and gender-matched volunteers without neurological disorders. Scans were acquired at rest and during production of continuous narrative speech and whispered speech. Speech was recorded during scan acquisition for offline quantification of voice breaks, pitch breaks, and percentage aperiodicity to assess correlations between treatment-related changes in rCBF and clinical improvement. Results demonstrated that speech-related responses in heteromodal sensory areas were significantly reduced in persons with ADSD, compared with volunteers, before the administration of BTX. Three to 4 weeks after BTX injection, speech-related responses were significantly augmented in these regions and in left hemisphere motor areas commonly associated with oral-laryngeal motor control. This pattern of responses was most strongly correlated with the objective measures of clinical improvement (decreases in the frequency of voice breaks, pitch breaks, and percentage aperiodicity). These data suggest a pathophysiological model for ADSD in which BTX treatment results in more efficient cortical processing of sensory information, making this information available to motor areas that use it to more effectively regulate laryngeal movements. PMID- 17077221 TI - Articulation rate and vowel space characteristics of young males with fragile X syndrome: preliminary acoustic findings. AB - PURPOSE: Increased speaking rate is a commonly reported perceptual characteristic among males with fragile X syndrome (FXS). The objective of this preliminary study was to determine articulation rate-one component of perceived speaking rate and vowel space characteristics of young males with FXS. METHOD: Young males with FXS (n = 38), developmental age (DA)-matched males (n = 21), and chronological age (CA)-matched males (n = 16) were audiotaped while engaged in spontaneous conversation and a picture-naming task. Articulation rate in syllables per second during intelligible utterances and vowel space area/dispersion measures were acoustically determined for each speaker. RESULTS: Males with FXS did not articulate significantly faster than CA-matched males. Area and dispersion of the acoustic vowel space also were similar between the 2 groups. Males with FXS, however, used significantly shorter utterances and had a tendency to pause less often than CA-matched males. In addition, males with FXS exhibited greater intraspeaker variability of formants associated with the vowel /a/. CONCLUSIONS: These preliminary findings suggest that articulation rate may not be a primary factor contributing to perceived speaking rate of males with FXS. Limitations of the study relative to speech production tasks and utterance intelligibility are discussed. PMID- 17077222 TI - Interspeaker variation in habitual speaking rate: additional evidence. AB - PURPOSE: The purpose of the present study was to test the hypothesis that talkers previously classified by Y.-C. Tsao and G. Weismer (1997) as habitually fast versus habitually slow would show differences in the way they manipulated articulation rate across the rate continuum. METHOD: Thirty talkers previously classified by Tsao and Weismer (1997) as having habitually slow (n = 15; 7 males, 8 females) and habitually fast (n = 15; 8 males, 7 females) articulation rates produced a single sentence at 7 different rates, using a magnitude production paradigm. Hence, the participants were not randomly assigned to conditions. RESULTS: Quadratic regression functions relating measured to intended articulation rates were all statistically significant, and most important, there were significant differences between the slow and fast groups in the y intercepts of the functions, for both males and females. CONCLUSIONS: This study provides a constructive replication of Tsao and Weismer (1997), showing a difference between slow and fast talkers with a new set of speech materials and in a new task. The findings appear to be consistent with a biological basis for intertalker rate differences. PMID- 17077223 TI - Patient safety in obstetrics and gynecology: the call to arms. PMID- 17077224 TI - Emergency contraception: politics and science move forward. PMID- 17077225 TI - Medical education: what's relevant, what's irrelevant, and what's missing. PMID- 17077226 TI - Pregnancy loss rates after midtrimester amniocentesis. AB - OBJECTIVE: The purpose of this study was to quantify the contemporary procedure related loss rate after midtrimester amniocentesis using a database generated from patients who were recruited to the First And Second Trimester Evaluation of Risk for Aneuploidy trial. METHODS: A total of 35,003 unselected patients from the general population with viable singleton pregnancies were enrolled in the First And Second Trimester Evaluation of Risk for Aneuploidy trial between 10 3/7 and 13 6/7 weeks gestation and followed up prospectively for complete pregnancy outcome information. Patients who either did (study group, n=3,096) or did not (control group, n=31,907) undergo midtrimester amniocentesis were identified from the database. The rate of fetal loss less than 24 weeks of gestation was compared between the two groups, and multiple logistic regression analysis was used to adjust for potential confounders. RESULTS: The spontaneous fetal loss rate less than 24 weeks of gestation in the study group was 1.0% and was not statistically different from the background 0.94% rate seen in the control group (P=.74, 95% confidence interval -0.26%, 0.49%). The procedure-related loss rate after amniocentesis was 0.06% (1.0% minus the background rate of 0.94%). Women undergoing amniocentesis were 1.1 times more likely to have a spontaneous loss (95% confidence interval 0.7-1.5). CONCLUSION: The procedure-related fetal loss rate after midtrimester amniocentesis performed on patients in a contemporary prospective clinical trial was 0.06%. There was no significant difference in loss rates between those undergoing amniocentesis and those not undergoing amniocentesis. LEVEL OF EVIDENCE: II-2. PMID- 17077227 TI - Adverse events detected by clinical surveillance on an obstetric service. AB - OBJECTIVE: Adverse events are adverse patient outcomes resulting from medical care. We performed this study to estimate the rate of adverse events and potential adverse events-errors that have a high likelihood of causing patient harm-occurring during obstetric care. METHODS: This was a prospective cohort study of an obstetric unit in a teaching hospital. We included patients admitted consecutively to the hospital. A trained observer monitored patients for 72 triggers, which were predefined occurrences deemed likely to indicate an actual or potential adverse event. When a trigger occurred, the observer captured information describing it. A five-person multidisciplinary team, including the observer, three physicians, and a hospital risk manager, judged whether the trigger represented an adverse event or potential adverse event. Adverse events were further characterized as preventable. RESULTS: The cohort included 425 patients; 47% were in active labor. We identified 110 triggers. Nine were considered adverse events (risk 2%, 95% confidence interval [CI] 1-4%, rate 0.8 events per 100 patient days), and six were preventable (risk 1%, 95% CI 0-3%, rate 0.5 events per 100 patient days). The remaining triggers included 14 potential adverse events (risk 3%, 95% CI 2-5%, rate 1.3 events per 100 patient days). No adverse event resulted in permanent disability or death. Adverse events and potential adverse events were most commonly "system" problems, such as unavailable staff or operating rooms, or poor fetal outcomes, such as trauma to the newborn. CONCLUSION: Serious adverse events occur infrequently on an obstetric service. However, important quality problems are common and should be targeted for improvement. LEVEL OF EVIDENCE: II-2. PMID- 17077228 TI - Pregnancy outcome after age 50. AB - OBJECTIVE: To evaluate pregnancy complications occurring after age 50. METHODS: We compared the pregnancy outcomes of women aged 50-64 years with those aged 45 49 years and with the general population. RESULTS: During 5 years from January 1, 1999, to June 30, 2004, 123 women aged 45 years and older gave birth. Fifty-five percent were nulliparous, 24 of 123 were aged 50-64 years, and 99 of 123 women were aged 45-49 years. All women older than age 50 conceived via in vitro fertilization with oocyte donation. For these 123 women, the overall mean gestational age at delivery was 37.6+/-2.6 weeks. The mean birth weight was 2,684+/-754 g, significantly lower than the general population, and the incidences of multifetal pregnancies, diabetes, and hypertension were high. Women aged 50 years and older were more likely to be hospitalized during pregnancy than women younger than 50 years (63% versus 22%, P<.001). Neonatal outcome was generally good. Women aged 50 years and older gave birth to significantly more low birth weight babies than those younger than age 50 years (61% versus 32%, P=.002). Gestational age and birth weight were both significantly lower for singletons and multiples in women older than age 50 years compared with those younger than age 50 years (gestational age of singletons 36.9 versus 38.4 weeks, P=.005; birth weight of singletons 2,694 versus 3,027 g, P=.019; gestational age of multiples 35.1 versus 36.4 weeks, P=.01; birth weight of multiples 1,976 versus 2,310 g, P=.038, respectively). CONCLUSION: Pregnant women aged 50-64 years have increased risks of preterm birth, low birth weight babies, diabetes mellitus, hypertension, and hospitalization. LEVEL OF EVIDENCE: II-2. PMID- 17077229 TI - Progesterone receptor modulator for emergency contraception: a randomized controlled trial. AB - OBJECTIVE: Compare the efficacy and adverse effects of CDB-2914, a new progesterone receptor modulator, to levonorgestrel for emergency contraception. METHODS: We performed a randomized, double-blinded noninferiority trial, enrolling healthy women seeking emergency contraception within 72 hours of unprotected intercourse. Participants were randomly assigned to receive a single dose of 50 mg of CDB-2914, plus a placebo 12 hours later or two doses of 0.75 mg of levonorgestrel taken 12 hours apart. Follow-up was scheduled 5 to 7 days after the expected onset of the next menstrual period. Posttreatment pregnancy was established by a positive urine test at follow-up and confirmed by quantitative serum beta-hCG. Daily diaries were used from the time of emergency contraception use until next menses to record adverse effects and sexual activity. RESULTS: Product efficacy was evaluable in 775 of CDB-2914 users and 774 of levonorgestrel users. Pregnancies occurred in 7 (0.9%, 95% confidence interval 0.2-1.6%) and 13 (1.7%, 95% confidence interval 0.8-2.6%) women, respectively. Based on the estimated cycle day of unprotected intercourse, 85% and 69% of anticipated pregnancies, respectively, were averted. Nausea was reported by a somewhat greater percentage of CDB-2914 than levonorgestrel users (29% compared with 24%, P=.03), but the distribution of other adverse effects was similar in both groups. Women in both groups experienced considerable variation in menstrual cycle length as compared with their reported individual normal cycle lengths. CONCLUSION: CDB 2914 is at least as effective as levonorgestrel in preventing pregnancies after unprotected intercourse and has a similar side effect profile. LEVEL OF EVIDENCE: I. PMID- 17077230 TI - Impact of increased access to emergency contraceptive pills: a randomized controlled trial. AB - OBJECTIVE: To assess how a strategy to maximize access to emergency contraceptive pills would affect rates of pregnancy and sexually transmitted infections. METHODS: Sexually active women, 14-24 years old, were randomly assigned to two methods of access to emergency contraceptive pills: increased access (two packages of pills dispensed in advance with unlimited resupply at no charge) or standard access (pills dispensed when needed at usual charges). Participants were followed for 1 year to assess incidence of pregnancy, gonorrhea, chlamydia, and trichomonas. RESULTS: The numbers of women enrolled in the increased and standard access groups were 746 and 744, respectively. More than 93% of participants completed a full year of follow-up. The incidence of pregnancy was similar in both groups (increased access group: 9.9/100 woman years, 95% confidence interval [CI] 7.7-12.6; standard access group: 10.5/100 woman years, 95% CI 8.2-13.2). Aggregate rates of gonorrhea, chlamydia, and trichomonas were also similar in the two groups (increased access group: 6.9/100 woman years, 95% CI 5.1-9.1; standard access group: 7.6/100 woman years, 95% CI 5.7-9.9). The increased access group used emergency contraceptive pills substantially more often and sooner after coitus than the standard access group. No other differences were noted between groups in self-reported measures of sexual behavior and contraceptive use. CONCLUSION: This intensive strategy to enhance access to emergency contraceptive pills substantially increased use of the method and had no adverse impact on risk of sexually transmitted infections. However, it did not show benefit in decreasing pregnancy rates. LEVEL OF EVIDENCE: II-1. PMID- 17077231 TI - Number of oral contraceptive pill packages dispensed, method continuation, and costs. AB - OBJECTIVE: To estimate the effect of the number of cycles of oral contraceptive pills (OCPs) dispensed per visit on method continuation, pill wastage, use of services, and health care costs. METHODS: We used paid claims data for 82,319 women dispensed OCPs through the California Family PACT (Planning, Access, Care, and Treatment) Program in January 2003 to examine contraceptive continuation and service use. RESULTS: Women who received 13 cycles at their first visit in January 2003 received 14.5 cycles over the course of 2003 compared with 9.0 cycles among women receiving three cycles at first visit. When client characteristics are controlled, women who received 13 cycles were 28% more likely to have OCPs on hand and twice as likely to have sufficient OCP cycles for 15 months of continuous use compared with women who received three cycles. Oral contraceptive pill wastage was higher among women initially dispensed 13 cycles (6.5% of the cycles dispensed) than among women who received three cycles (2% of cycles). Despite having one fewer clinician visit, women dispensed 13 cycles were more likely to receive Pap and Chlamydia tests and less likely to have a pregnancy test than women initially dispensed fewer cycles. Over the course of the year, Family PACT paid 99 US dollars more for women who received three cycles and 44 US dollars more for women who received only one cycle than it did for women who received 13 cycles at their first visits of 2003. CONCLUSION: Dispensing a year's supply of OCP cycles to women is associated with higher method continuation and lower costs than dispensing fewer cycles per visit. LEVEL OF EVIDENCE: II-2. PMID- 17077232 TI - Sample bias among women with retained DNA samples for future genetic studies. AB - OBJECTIVE: To evaluate whether women who agree to future use of their biologic specimens for genetic studies reflect the larger study population from which they are derived. METHODS: Women were questioned as to the future disposition of their maternal and fetal DNA samples upon enrollment in a multicenter, observational study originally designed to identify factor V Leiden mutation carriers and prospectively ascertain the estimated rate of pregnancy-related venous thromboembolism and adverse pregnancy outcome. Univariate and multivariate analyses was carried out on the 5,003 of 5,188 enrolled women who indicated their desire regarding future disposition of their DNA samples. RESULTS: Among these 5,003 women, 20.1% desired that their samples be discarded and not available for future genetic studies. Multivariate analysis demonstrated that women who agreed to subsequent use of samples were less likely African-American (odds ratio [OR] 0.6, 95% confidence interval [CI] 0.4-0.7) or Hispanic (OR 0.4, 95% CI 0.3-0.5), and more likely to use tobacco (OR 1.2, 95% CI 1.0-1.6) than those who desired that their samples be discarded. CONCLUSION: Genetic samples from women agreeing to their use in a sample repository may not be representative of the index study cohort. This should be considered in their subsequent interpretation and generalizability. LEVEL OF EVIDENCE: III. PMID- 17077233 TI - Patient refusal of emergency cesarean delivery: a study of obstetricians' attitudes in Europe. AB - OBJECTIVE: To compare the attitudes of a large sample of obstetricians from eight European countries toward a competent woman's refusal to consent to an emergency cesarean delivery for acute fetal distress. METHODS: Obstetricians' attitudes in response to a hypothetical clinical case were surveyed through an anonymous, self administered questionnaire. The sample included 1,530 obstetricians (response rate 77%) from 105 maternity units (response rate 70%) in eight countries: France, Germany, Italy, Luxembourg, Netherlands, Spain, Sweden and the United Kingdom. RESULTS: In every country, the majority of obstetricians would keep trying to persuade the woman, telling her that failure to perform cesarean delivery might result in the fetus surviving with disability, or even that her own life might be endangered. In Spain, France, Italy, and, to a lesser extent, Germany and Luxembourg, a consistent proportion of physicians would seek a court order to protect fetal welfare or avoid possible legal liability or both. In the United Kingdom, Sweden, and Netherlands, several respondents (59%, 41%, and 37%, respectively) would accept the woman's decision and assist vaginal delivery. Only a small minority (from 0 in the United Kingdom to 10% in France) would proceed with cesarean delivery without a court order. CONCLUSION: Case law arising from a few countries (United States, Canada, and the United Kingdom) and professional guidelines favoring women's autonomy have not solved the underlying ethical conflict, and in Europe acceptance of a woman's right to refuse cesarean delivery, at least in emergency situations, is not uniform. Differing attitudes between obstetricians from the eight countries may reflect diverse legal and ethical environments. LEVEL OF EVIDENCE: III. PMID- 17077234 TI - Controlled-release misoprostol vaginal insert in parous women for labor induction: a randomized controlled trial. AB - OBJECTIVE: To assess the ability of a controlled-release misoprostol vaginal insert to induce labor using dose reservoirs of 25, 50, 100, and 200 microg. METHODS: This double-blind, dose ranging, randomized study was carried out in parous women requiring induction of labor at term. Each woman was randomly assigned to receive a single misoprostol vaginal insert that could remain in place for up to 24 hours and was removed for onset of active labor, an adverse event, or having reached 24 hours in situ. The primary outcome measure was time from insertion of the misoprostol vaginal insert to vaginal delivery of the neonate. RESULTS: A total of 124 women participated in the study. The median time to vaginal delivery was 27.5, 19.1, 13.1, and 10.6 hours for the 25-, 50-, 100-, and 200-microg doses, respectively. The percentage of women who delivered vaginally within 12 hours was 9%, 14%, 47%, and 53% (P<.001 using the 25-microg group as the comparator) and within 24 hours was 42%, 79%, 81%, and 70% (P=.003). Uterine hyperstimulation syndrome occurred in one woman who received the 25-mug, two women who received the 100-microg, and three women who received the 200 microg dose reservoirs. CONCLUSION: Misoprostol vaginal inserts effectively induced labor in pregnant parous women at term. LEVEL OF EVIDENCE: I. PMID- 17077235 TI - Effect on neonatal outcomes in gestational hypertension in twin compared with singleton pregnancies. AB - OBJECTIVE: We tested the hypothesis that gestational hypertension may have a more benign effect on neonatal outcomes in twin compared with singleton pregnancies, because the elevated blood pressure in twin pregnancies may partly or merely reflect the extra demand for blood supply. METHODS: A retrospective cohort study of 102,988 twin and 5,523,797 singleton live births using the U.S. birth cohort linked birth and infant death data sets, 1998-2000. Main outcomes are relative risks (RRs) of adverse neonatal outcomes: preterm birth, intrauterine growth restriction (less than the third percentile), low 5-minute Apgar score (less than 4), and neonatal death comparing gestational hypertensive with no-event healthy pregnancies for twins and singletons. RESULTS: For singletons, crude RRs (95% confidence intervals) comparing gestational hypertensive with healthy pregnancies were 2.23 (2.20-2.25) for preterm birth (17.4 compared with 7.8%), 2.49 (2.45 2.53) for intrauterine growth restriction (7.4 compared with 3.0%), 1.33 (1.21 1.45) for low 5-minute Apgar score (2.6 compared with 2.0 per 1,000), and 1.07 (0.96-1.19) for neonatal death (1.9 compared with 1.8 per 1,000), respectively. For twins, the corresponding RRs were much lower or showed reversed associations: 1.21 (1.19-1.24) (63.6 compared with 52.4%), 1.04 (0.98-1.11) (16.4 compared with 16.4%), 0.32 (0.23-0.46) (4.1 compared with 12.7 per 1,000), and 0.21 (0.14-0.30) (3.6 compared with 17.2 per 1,000), respectively. The adjusted odds ratios showed a similar risk pattern in twin compared with singleton pregnancies after controlling for maternal race, age, education, marital status, parity, smoking, alcohol use, perinatal care use, and mode of delivery. CONCLUSION: Gestational hypertension has a much more benign effect on neonatal outcomes in twin compared with singleton pregnancies. There might be a need for twin- or multiple fetus specific recommendations for hypertension management in pregnancy, but further interventional studies are needed to test the hypothesis. LEVEL OF EVIDENCE: II 2. PMID- 17077236 TI - Iron intake and risk of ovulatory infertility. AB - OBJECTIVE: To evaluate whether iron supplement use or greater intake of total, heme and nonheme iron is associated with lower risk of ovulatory infertility. METHODS: We conducted a prospective cohort study among 18,555 married, premenopausal women without a history of infertility who attempted a pregnancy or became pregnant between 1991 and 1999 (mean baseline age+/-standard deviation 32.6+/-3.6). Diet was assessed twice during follow-up and prospectively related to the incidence of infertility due to ovulatory disorder. RESULTS: During the 8 years of follow-up, 438 women reported infertility due to ovulatory disorder. Women who consumed iron supplements had a significantly lower risk of ovulatory infertility than women who did not use iron supplements (relative risk 0.60, 95% confidence interval 0.39-0.92), after adjusting for potential confounders. Total nonheme iron intake, primarily consumed as multivitamins and iron supplements, was inversely associated with the risk of infertility (relative risk Quintile 1 compared with 5, 95% confidence interval 0.39-0.92; P, trend .005.) Heme iron intake was unrelated to ovulatory infertility in multivariable adjusted analyses. CONCLUSION: Consumption of iron supplements and nonheme iron from other sources may decrease the risk of ovulatory infertility. LEVEL OF EVIDENCE: II-2. PMID- 17077237 TI - Pregnancy weight gain and risk of neonatal complications: macrosomia, hypoglycemia, and hyperbilirubinemia. AB - OBJECTIVE: To examine whether pregnancy weight gains outside the Institute of Medicine (IOM) recommendations and rates of maternal weight gain are associated with neonatal complications. METHODS: In a cohort of 45,245 women who delivered singletons at Kaiser Permanente Medical Care Program Northern California in 1996 1998 and who did not have gestational diabetes as of 24-28 weeks of gestation, we conducted a nested case-control study with three case groups: macrosomia (birth weight more than 4,500 g, n=391), neonatal hypoglycemia (plasma glucose less than 40 mg/dL, n=328), and hyperbilirubinemia (serum bilirubin 20 mg/dL or more, n=432) and one control group (n=652). Medical records were reviewed to ascertain the woman's prepregnancy and predelivery weight. RESULTS: Adjusting for age, race ethnicity, parity, plasma glucose screening value, and difference in weeks between delivery and time when last weight was measured, women who gained more than recommended by the IOM were three times more likely to have an infant with macrosomia (odds ratio [OR] 3.05, 95% confidence interval [CI] 2.19-4.26), and nearly 1.5 times as likely to have an infant with hypoglycemia (OR 1.38, 95% CI 1.01-1.89), or hyperbilirubinemia (OR 1.43, 95% CI 1.06-1.93) than women whose weight gain was in the recommended range. Women who gained less than the IOM recommendations were less likely than women in the recommended range to have an infant with macrosomia (OR 0.38, 95% CI 0.20-0.70), but equally likely to have an infant with hypoglycemia or hyperbilirubinemia. Similar results were obtained using other means of categorizing weight gain during pregnancy. CONCLUSION: Maternal weight gain above the IOM recommendations was associated with an increased risk of the outcomes studied. LEVEL OF EVIDENCE: II-2. PMID- 17077238 TI - Adhesion-related bowel obstruction after hysterectomy for benign conditions. AB - OBJECTIVE: To evaluate the occurrence of small bowel obstruction after hysterectomy. METHODS: Analysis of 326 cases of women who were admitted with a diagnosis of small bowel obstruction during the period 1998-2005. Among cases with small bowel obstruction after hysterectomy for benign conditions, we evaluated the type and technique of hysterectomy and whether the parietal peritoneum was sutured at the completion of the procedure. RESULTS: The main causes of bowel obstruction were intra-abdominal adhesions (41.9%) and abdominal malignancy (40.1%). After excluding oncologic cases, we found that, of 135 cases of adhesion-related small bowel obstruction, gynecologic operations played the largest role in the occurrence of bowel obstruction (n=68, 50.4%). Among all gynecologic operations for benign conditions, total abdominal hysterectomy (TAH) was the most common cause of small bowel obstruction (13.6 per 1,000 TAHs). We did not encounter small bowel obstruction after laparoscopic supracervical hysterectomy. The reduction in absolute risk of small bowel obstruction from TAH to laparoscopic supracervical hysterectomy is 13.6 per 1,000 cases; 73 patients would undergo laparoscopic supracervical hysterectomy to prevent one small bowel obstruction. The median interval between TAH and small bowel obstruction was 4 years. The adhesions were adherent to the previous laparotomy incision in 27 cases (75%) and to the vaginal vault in nine cases (25%). Peritoneal closure was not associated with small bowel obstruction. CONCLUSION: Hysterectomy plays a major role in the occurrence of adhesion-related small bowel obstruction. Closure of the parietal peritoneum does not contribute to the occurrence of adhesion related small bowel obstruction, and small bowel obstruction rarely occurs after laparoscopic supracervical hysterectomy. LEVEL OF EVIDENCE: II-3. PMID- 17077239 TI - Evaluation of adnexal masses with three-dimensional ultrasonography. AB - OBJECTIVE: To estimate whether three-dimensional ultrasonography and three dimensional power Doppler investigation can contribute to the differentiation between benign and malignant ovarian masses. METHODS: Women scheduled for surgical treatment of an adnexal mass were included in a multicenter prospective study. All women underwent two-dimensional and three-dimensional ultrasonographic examination in the week before surgery. All parameters were compared in women with benign tumors, borderline tumors, and malignant tumors using receiver operating characteristic analysis and likelihood ratios. RESULTS: We included 181 women; 144 had a benign mass, 26 had a malignancy, and 11 had a borderline tumor. At three-dimensional ultrasonography, the most striking difference was found in the presence of central vessels in an adnexal mass. Central vessels assessed by three-dimensional ultrasonography were present in 15% (21 of 144) of the benign masses, 69% (18 of 26) of the malignant masses, and 27% (3 of 11) of the masses of borderline malignancy. The likelihood ratios for presence of central vessels for a mass being malignant and/or borderline was 4.9 (95% confidence interval 2.1 12). Mean gray index and flow index were also significantly different between the groups, but other features were not. CONCLUSION: The central localization of vessels in an adnexal mass, as observed by three-dimensional ultrasonography, the mean gray index, and the flow index are potentially important parameters for distinguishing benign from malignant adnexal masses. LEVEL OF EVIDENCE: II-2. PMID- 17077240 TI - Ovarian cancer screening in women with a family history of breast or ovarian cancer. AB - OBJECTIVE: To evaluate positive predictive values of CA 125 or transvaginal ultrasonography screening for ovarian cancer according to family history of breast or ovarian cancer. METHODS: In the screening arm of a randomized controlled trial of screening compared with usual care, 28,460 women with family history data received baseline and annual CA 125 and transvaginal ultrasonography examinations. We analyzed CA 125 and transvaginal ultrasonography results from the first four rounds of screening. We classified women as average (n=22,687), moderate (n=2,572), or high (n=2,163) risk based on family history, or high risk due to a personal history of breast cancer (n=1,038). Cancers were identified by active follow-up of women with abnormal screening results and annual questionnaires. We calculated positive predictive values for screening combinations. RESULTS: Similar proportions (4.8-5.0%) of women in each group had abnormal screening results. Higher-risk women were more likely than lower-risk women to undergo biopsy after a positive screen. Screening identified 43 invasive ovarian cancers. The positive predictive values for abnormal screening results were 0.7% in average-risk, 1.3% in moderate-risk, and 1.6% in high-risk groups; one ovarian cancer occurred among the breast cancer survivors. The positive predictive values for postbaseline abnormal screening results were also higher in the higher-risk groups. The positive predictive values did not significantly differ across risk groups. CONCLUSION: Probabilities of abnormal annual CA 125 and transvaginal ultrasonography screens were similar across groups based on family history of breast or ovarian cancer. However, ovarian cancer was more likely to be diagnosed after an abnormal screening result among women at higher family history-based risk than among women at lower risk. LEVEL OF EVIDENCE: I. PMID- 17077241 TI - Causes of chronic vaginitis: analysis of a prospective database of affected women. AB - OBJECTIVE: To compare women with different chronic vaginal symptoms with a wide variety of sociodemographic, health, behavioral, and psychosocial characteristics. METHODS: Serially recruited subjects answered a questionnaire that asks about demographic information and symptoms and measures depression and stress scores. Patients underwent a standardized history, physical examination, and laboratory examination. Patients with recurrent vulvovaginal candidiasis, vulvar vestibulitis syndrome, desquamative inflammatory vaginitis, physiologic leukorrhea, and other diagnoses were compared with one another. Chi-square tests and one-way analysis of variance with Tukey honestly significant difference (HSD) post hoc analyses were used for categorical and continuous data analysis. RESULTS: Two hundred patients were enrolled in this study. The most common diagnoses were contact dermatitis (21%), recurrent vulvovaginal candidiasis (20.5%), atrophic vaginitis (14.5%), and vulvar vestibulitis syndrome (12.5%); 18% of women had 2 or more diagnoses. In the overall study sample, the mean age was 38.4 years, 78% were white, and 55% were college educated. Sixty-two percent had symptoms for over a year. Desquamative inflammatory vaginitis patients were older and less likely to be menstruating. Those with vulvar vestibulitis syndrome had more frequent complaints of dyspareunia. Recurrent vulvovaginal candidiasis patients felt that their symptoms had the greatest negative impact on work and social life. There were high rates of psychiatric disorder (43.5%), atopic disease (42.5%), and pain syndrome (56%) in all groups. CONCLUSION: Women with chronic vaginal symptoms have a variety of diagnoses, most of them noninfectious. LEVEL OF EVIDENCE: II-3. PMID- 17077242 TI - Comparison of serum markers in first-trimester down syndrome screening. AB - OBJECTIVE: To estimate patterns of total hCG and inhibin A levels in the late first trimester of Down syndrome pregnancies, compare them with that of free beta hCG, and assess screening performance of these markers individually and in combination with pregnancy-associated plasma protein-A (PAPP-A) and nuchal translucency. METHODS: Seventy-nine matched case-control sets of maternal serum samples (each Down syndrome case matched to 5 controls) from 11 through 13 completed weeks of gestation were taken from the sample bank of the First and Second Trimester Evaluation of Risk Consortium, a population-based study, and assayed for levels of free beta-hCG, total hCG, and inhibin A. Distribution characteristics and correlations of the multiples of the median values were estimated in cases and controls. Screening performance for each marker, alone and in combination with PAPP-A, nuchal translucency, and maternal age, was calculated. RESULTS: Median multiples of the median levels of free beta-hCG, total hCG, and inhibin A in cases were more elevated as gestation increased from 11 to 13 weeks, with univariate detection rates of 31%, 23%, and 29%, respectively, at a 5% false-positive rate. At 12 weeks, the multivariate detection rates at a 5% false-positive rate for nuchal translucency and PAPP-A (with maternal age) with either free beta-hCG, total hCG, or inhibin A were 84%, 83%, and 85%, respectively. The improvement in performance from nuchal translucency and PAPP-A to any of the three-marker tests was significant, while performance of any of the three-marker combinations was not significantly different from each other. CONCLUSION: Although levels of free beta-hCG in affected pregnancies were higher earlier than the levels of either total hCG or inhibin A, there was no significant difference in screening performance when either of the three markers was used with nuchal translucency and PAPP-A at 11-13 weeks of pregnancy. LEVEL OF EVIDENCE: II-2. PMID- 17077243 TI - Associations of physical activity and inactivity before and during pregnancy with glucose tolerance. AB - OBJECTIVE: To investigate associations of physical activity and television viewing before and during pregnancy with risk of gestational diabetes mellitus (GDM) and abnormal glucose tolerance, the combination of GDM with less severe impaired glucose tolerance. METHODS: We assessed duration and intensity of physical activity and time spent viewing television both before and during pregnancy among 1,805 women enrolled in Project Viva, a cohort study in eastern Massachusetts. We identified 1,493 (83%) women with normal glucose tolerance and 312 (17%) with abnormal glucose tolerance, including 91 (5%) with GDM based on clinical glucose tolerance test results. RESULTS: After adjustment for age, race or ethnicity, history of GDM, family history of diabetes, and prepregnancy body mass index, our data suggest that women who engaged in any vigorous physical activity in the year before pregnancy experienced a reduced risk of GDM (odds ratio [OR] 0.56, 95% confidence interval [CI] 0.33-0.95) and abnormal glucose tolerance (OR 0.76, 95% CI 0.57-1.00). Women who reported vigorous activity before pregnancy and light-to-moderate or vigorous activity during pregnancy appeared to have a lower risk of both GDM (OR 0.49, 95% CI 0.24-1.01) and abnormal glucose tolerance (OR 0.70, 95% CI 0.49-1.01) compared with women reporting these activities in neither time period. Walking and total physical activity provided modest benefits. We observed no association of television viewing before or during pregnancy with risk of GDM or abnormal glucose tolerance. CONCLUSION: Physical activity, especially vigorous activity before pregnancy and at least light-to-moderate activity during pregnancy, may reduce risk for abnormal glucose tolerance and GDM. LEVEL OF EVIDENCE: II-2. PMID- 17077244 TI - Improved survival with an intact DNA mismatch repair system in endometrial cancer. AB - OBJECTIVE: To correlate survival and surgical-pathologic factors with DNA mismatch repair status in patients with endometrial cancer. METHODS: Specimens from 336 patients with endometrial cancer were used to create a tissue microarray. Immunohistochemistry with antibodies against the mismatch repair genes MLH1, MSH2, MSH6, and PMS2 were used to stain the tissue microarray. Clinical, pathologic, and survival data were collected and correlated with the immunohistochemistry results. RESULTS: Mismatch repair deficiency was seen in 29% (84 of 294) of cases. Correlation was noted between lack of expression of MLH1 and an increased risk for lymphvascular space involvement (32% versus 21%, P=.05) and cervical involvement (26% versus 14%, P=.02). Lack of expression of either MLH1 or MSH2 was associated with thinner patients (85% had a body mass index less than 40 versus 73% of patients with normal expression, P=.02), as well as with the absence of a history of previous primary malignancy (0 verus 13 cases [4%], P=.023). The estimated disease-free survival is 88%; despite a small number of recurrences, there was a nonsignificant improvement in disease-free survival in tumors with an intact mismatch repair system (P=.1). Significantly improved disease-free survival was seen in patients with normal MLH1 and MSH2 expression compared with those with abnormal expression (92% versus 81%, P=.035). CONCLUSION: Defects in DNA mismatch repair in endometrial cancer is correlated with negative prognostic factors and worse progression-free survival (without a difference in overall survival) compared with tumors with an intact mismatch repair system. LEVEL OF EVIDENCE: II-3. PMID- 17077245 TI - Tenure of academic chairs in obstetrics and gynecology: a 25-year perspective. AB - OBJECTIVE: To assist in predicting future leadership needs, this longitudinal study examines turnover and net retention rates among chairs at university obstetrics and gynecology departments between 1981 and 2005. METHODS: A database of appointment dates and tenure of chairs at each of 125 Association of American Medical Colleges-approved United States medical schools was collated using membership listings from the Association of Professors of Gynecology and Obstetrics and from the Council of University Chairs in Obstetrics and Gynecology. Complete data from 118 departments were confirmed by selective correspondence at individual departments and further review by the investigators. RESULTS: A total of 260 individuals (232 men, 28 women) became new chairs between 1981 and 2005. The annual turnover rate increased gradually from 6.0% to 12.7%. Five-year net retention rates remained steady between 1982 and 1997 but dropped after 1997 (85.6% compared with 63.2%; P=.03). A chair's tenure ranged widely (1 to 23 years; median 8 years), regardless of gender or school type, size, or location. Approximately one half of interim chairs became permanent chairs, usually at their own institution. The number of new women chairs increased from none in 1981 to 17 (15.2% of total chairs) in 2005. CONCLUSION: Academic chair positions in obstetrics and gynecology experienced a doubling in annual turnover rates, while retention rates declined. The proportion of chairs occupied by women increased progressively. LEVEL OF EVIDENCE: II-2. PMID- 17077246 TI - An effective pressure pack for severe pelvic hemorrhage. AB - OBJECTIVE: To evaluate our experience with the "pelvic pressure pack," a surgical technique for controlling posthysterectomy bleeding. METHODS: This is an observational descriptive report of cases collected by the authors during the years 1968-2006. Packs were constructed of various materials (eg, pillow cases, gauze sheets, plastic X-ray cassette drapes, or orthopedic stockings) filled with gauze rolls introduced abdominally and exiting the vagina. RESULTS: We report 11 new cases (10 obstetric and one gynecologic). Massive red blood cell transfusion and coagulopathy occurred in all cases. The pelvic pressure pack successfully controlled bleeding in 82% (9 of 11) of cases. Postoperative febrile morbidity occurred in most cases. There were no maternal deaths. CONCLUSION: In the contemporary management of posthysterectomy bleeding, the pelvic pressure pack appears to be a valuable surgical option, affording correction of coagulopathy and further stabilization. Given near-universal postoperative febrile morbidity in our series, prophylactic broad-spectrum antibiotic therapy should be strongly considered. We believe all obstetrician-gynecologists should be familiar with this simple, potentially life-saving technique. LEVEL OF EVIDENCE: III. PMID- 17077247 TI - A complex ovarian cyst. AB - BACKGROUND: Acute pelvic pain in women is a common diagnostic dilemma encountered by the gynecologist. We describe a case of acute pelvic pain with an unusual etiology. CASE: A 40-year-old multipara presented within hours of the onset of acute pelvic pain. She was treated conservatively with analgesics after computed tomography of the abdomen and pelvis revealed a ruptured ovarian cyst. Upon return to the emergency room, she was admitted to a gynecologic service for pain control. The etiology of her acute pelvic pain eventually declared itself. CONCLUSION: The differential diagnosis of acute pelvic pain represents a variety of organ systems. PMID- 17077248 TI - Can you find the source of her pain? AB - BACKGROUND: The features of abdominal pain in this gravid patient mimicked more common diagnoses like preterm labor, chorioamnionitis, and appendicitis. CASE: A 40-year-old multipara presented at 30 weeks and 6 days with abdominal pain. The cause was not discovered until the time of cesarean delivery several days after admission. CONCLUSION: This common gynecologic problem can precipitate severe problems in a pregnant woman and should be considered part of the diagnosis in pregnant patients presenting with pain. PMID- 17077249 TI - Don't do anything rash! AB - BACKGROUND: Peripartum and postpartum dermatologic lesions may or may not be an indicator of morbidity or mortality for either the fetus or mother. CASE: Postpartum dermatologic lesions occurred in a 41-year-old multipara, 1 week after a repeat cesarean delivery for severe intrauterine growth restriction and nonreassuring fetal testing. The diagnosis, using histopathologic techniques, and the pharmacologic management are discussed. CONCLUSION: This case raises intriguing questions regarding prompt diagnosis, treatment, and the counseling of patients with dermatologic lesions with respect to current and future pregnancy outcomes. PMID- 17077250 TI - The honeymoon is over. AB - BACKGROUND: The importance of a broad differential diagnosis is highlighted by a complex patient case. CASE: A young woman returned from her honeymoon complaining of activity-limiting pain. The evaluation revealed multiple constitutional symptoms, a breast mass, a large pelvic mass, pulmonary abnormalities, an external iliac venous thrombosis, and lytic bone lesions. Biopsies from several sites revealed no evidence of neoplasia. CONCLUSION: Consideration of potential etiologies outside of one's practice specialty may be necessary to make a correct diagnosis. PMID- 17077251 TI - Evidence-based strategies for implementing guidelines in obstetrics: a systematic review. AB - OBJECTIVE: To estimate effective strategies for implementing clinical practice guidelines in obstetric care and to identify specific barriers to behavior change and facilitators in obstetrics. DATA SOURCES: The Cochrane Library, EMBASE, and MEDLINE were consulted from January 1990 to June 2005. Additional studies were identified by screening reference lists from identified studies and experts' suggestions. METHODS OF STUDY SELECTION: Studies of clinical practice guidelines implementation strategies in obstetric care and reviews of such studies were selected. Randomized controlled trials, controlled before-after studies, and interrupted time series studies were evaluated according to Effective Practice and Organization of Care criteria standards. TABULATION, INTEGRATION, AND RESULTS: Studies were reviewed by two investigators to assess the quality and the efficacy of each strategy. Discordances between the two reviewers were resolved by consensus. In obstetrics, educational strategies with medical providers are generally ineffective; educational strategies with paramedical providers, opinion leaders, qualitative improvement, and academic detailing have mixed effects; audit and feedback, reminders, and multifaceted strategies are generally effective. These findings differ from data on the efficacy of clinical practice guidelines implementation strategies in other medical specialties. Specific barriers to behavior change in obstetrics and methods to overcome these barriers could explain these differences. The proportion of effective strategies is significantly higher among the interventions that include a prospective identification of barriers to change compared with standardized interventions. CONCLUSION: Prospective identification of efficient strategies and barriers to change is necessary to achieve a better adaptation of intervention and to improve clinical practice guidelines implementation. In the field of obstetric care, multifaceted strategy based on audit and feedback and facilitated by local opinion leaders is recommended to effectively change behaviors. PMID- 17077252 TI - Antenatal herpes serologic screening: an appraisal of the evidence. AB - OBJECTIVE: Calls for universal antenatal type-specific herpes simplex virus (HSV) screening to prevent neonatal herpes have recently increased and would affect the four million pregnant women and their partners annually in the United States. We undertook this review to assess the appropriateness of such screening, making relevant comparisons to established antenatal human immunodeficiency virus (HIV) and hepatitis B virus (HBV) screening programs. DATA SOURCES: We conducted a full PubMed and bibliographic search for relevant literature in English available from 1966 through February 2006, using the terms "genital herpes," "neonatal herpes," "decision analysis" or "cost-effectiveness analysis," and "herpes and pregnancy" or "antenatal herpes screening." Comparison literature was obtained by replacing "herpes" with "HBV" or "HIV". METHODS OF STUDY SELECTION: We appraised antenatal type-specific HSV screening using well-established criteria for a good screening program, which we articulated as questions. Of 455 articles we selected those that addressed at least one of the questions and were pertinent to the U.S. population. TABULATION, INTEGRATION, AND RESULTS: We found that neonatal HSV is rare and its incidence is imprecisely defined. There is a lack of evidence supporting the effectiveness of interventions to prevent maternal acquisition of new infection in late pregnancy, which accounts for 60-80% of neonatal herpes. The consequences of universal screening are incompletely understood but include the potential for unnecessary cesarean deliveries and medical treatment, maternal psychosocial stress, and discord among partners. The available evidence indicates universal screening is not cost-effective. In contrast, antenatal HIV and HBV screening programs better satisfy accepted criteria for screening. CONCLUSION: On the basis of this appraisal, universal antenatal type-specific HSV screening to prevent neonatal herpes does not adequately satisfy criteria of a good screening program, and we recommend against its adoption. PMID- 17077253 TI - Duration of intrapartum prophylaxis for neonatal group B streptococcal disease: a systematic review. AB - OBJECTIVE: To examine published evidence regarding duration of intrapartum antibiotic prophylaxis administered to pregnant women colonized with group B Streptococcus (GBS) to reduce infant colonization with GBS and to prevent early onset GBS sepsis. DATA SOURCES: A search was conducted in The Cochrane Central Register of Controlled Trials (The Cochrane Library, Issue 1, 2006), MEDLINE (1966 to January 2006), EMBASE (1980 to January 2006), CINAHL (1982 to January 2006), and in protocols and guidelines of the Centers for Disease Control and Prevention, American Academy of Pediatrics, and American College of Obstetrics and Gynecology. METHODS OF STUDY SELECTION: All randomized controlled trials and observational studies in which duration of intrapartum antibiotic prophylaxis is reported relative to subsequent neonatal GBS colonization or sepsis were considered. Case series and study designs using historical cohorts or controls for comparison were excluded. TABULATION, INTEGRATION, AND RESULTS: Three prospective cohort studies and one case-control study met inclusion criteria. Heterogeneity of study design and assembly of cohorts precluded meta-analysis. A systematic review of the individual studies was performed. All studies were rated as fair or poor validity with regard to their ability to evaluate duration of intrapartum prophylaxis and transmission of GBS to the newborn. All 4 studies were largely composed of women with existing risk factors for GBS disease of the newborn. One study supported more than 1 hour of prophylaxis, two studies supported more than 2 hours of prophylaxis, and one was inconclusive. CONCLUSION: Despite unequivocal clinical guidelines recommending at least 4 hours of intrapartum antibiotic prophylaxis, there are no well-designed studies examining duration of intrapartum antibiotic prophylaxis for prevention of early-onset GBS disease of the newborn. We recommend continuing to initiate intrapartum prophylaxis according to the American College of Obstetricians and Gynecologists guidelines; however, the transmission of GBS to neonates exposed to less than 4 hours of intrapartum prophylaxis and their subsequent management require further study. PMID- 17077254 TI - Patient safety in obstetrics and gynecology: an agenda for the future. AB - The effect of medical errors and unsafe systems of care has had a profound effect on the practice of obstetrics and gynecology. From 1975 to 2000, medical malpractice costs for obstetrician-gynecologists have risen nearly four-fold higher than that of other medical costs. In addition, it has been estimated that defensive medicine may cost society $80 billion per year. Most importantly, many obstetrician-gynecologists are frustrated and seem to be abandoning the parts of their practice they perceive to put them at higher liability risk. This article discusses other medical specialty society efforts that have been successful in addressing the area of patient safety. Efforts to better track quality outcomes has been initiated by the American College of Surgeons through the National Surgical Quality Improvement Project, and the American Society of Anesthesiologists has demonstrated both dramatically improved outcomes and reduced liability costs through a concerted patient safety effort. The author proposes changes in four areas to specifically address patient safety in obstetrics and gynecology, including: the development of reliable and reproducible quality control measures (and a system to track them); national closed claim reviews to better understand and address the most important safety and liability areas for obstetrician-gynecologists; work prospectively with pharmaceutical and surgical device manufacturers to develop innovative new products that would increase the likelihood of safe outcomes; and create a culture of safety in obstetrics and gynecology by incorporating safety education into all levels of training. PMID- 17077255 TI - Images of American sexuality in debates over nonprescription access to emergency contraceptive pills. AB - The debate over emergency contraceptive pill access in the United States revolves around speculations about Americans' sexual lives. The recently released internal U.S. Food and Drug Administration (FDA) memo that expresses fears that adolescents will form "sex-based cults" around emergency contraceptive pills echoes arguments made against the nonprescription switch at the 2003 FDA hearings. In these hearings, opponents argued that nonprescription access would lead to adolescent promiscuity and disease transmission and that adult predators would use the drug to facilitate the sexual abuse of young women. In contrast, proponents of expanded access to emergency contraceptive pills overwhelmingly portrayed their model user as a responsible adult who experiences a torn condom during consensual sex. These imaginations of American sexuality are tied to competing models of the role of medical providers in women's sexual decision making. Opponents of the nonprescription switch argued that women need a learned intermediary, not only to determine their need for emergency contraception, but also to educate them about proper sexual behavior and protect them from abuse. Proponents advocated putting more responsibility for sexual health decision making in the hands of women, not doctors, and complained about the moralizing scrutiny of medical providers. In the absence of nonprescription access to emergency contraception, advance prescription of emergency contraceptive pills can ensure that contraceptive education is not tied to a specific sexual act and therefore not perceived as a judgment about women's sexual decisions. However, advance prescription does not help women who lack access to health care or women who make sexual and contraceptive decisions without consulting physicians. PMID- 17077256 TI - Revisiting pharmacists' refusals to dispense emergency contraception. AB - Pharmacists' refusals to fill prescriptions for emergency contraceptives for reasons of conscience have contributed to a national debate regarding the permissibility of such actions. Some in the medical community assert that pharmacists ought not to refuse to dispense emergency contraceptives on this basis. Three lines of argument have become prominent in defense of that position: 1) the professional status of pharmacists does not allow for refusal to dispense legitimately written prescriptions, 2) the medical facts regarding the mechanism of action of emergency contraception are often misunderstood, misrepresented, or both, and 3) refusals by pharmacists to fill legitimate prescriptions undermine patient care. In this commentary, these arguments are rejected as missing the central point of the issue, which is that pharmacists are autonomous, moral agents who are accountable for their choices and entitled-within limits-to decide in which activities they will participate. Pharmacists' professionalism is defended, their responsibilities in the provision of drug therapy are set forth in the context of pharmaceutical care, and these lead to the conclusion that pharmacists' refusals may be ethically justified. There are important limits on how pharmacists may respond when they are being asked to participate in actions they find morally objectionable. Notably, they must ensure that these prescriptions are filled by someone else in a timely manner and must refrain from any abusive or demeaning treatment of patients, as summed up in our Principle of Conscientious Refusal to Dispense. PMID- 17077257 TI - Thyroid disease in pregnancy. AB - Thyroid testing during pregnancy should be performed on symptomatic women or those with a personal history of thyroid disease. Overt hypothyroidism complicates up to 3 of 1,000 pregnancies and is characterized by nonspecific signs or symptoms that are easily confused with complaints common to pregnancy itself. Physiologic changes in serum thyroid-stimulating hormone (TSH) and free thyroxine (T(4)) related to pregnancy also confound the diagnosis of hypothyroidism during pregnancy. If the TSH is abnormal, then evaluation of free T(4) is recommended. The diagnosis of overt hypothyroidism is established by an elevated TSH and a low free T(4). The goal of treatment with levothyroxine is to return TSH to the normal range. Overt hyperthyroidism complicates approximately 2 of 1,000 pregnancies. Clinical features of hyperthyroidism can also be confused with those typical of pregnancy. Clinical hyperthyroidism is confirmed by a low TSH and elevation in free T(4) concentration. The goal of treatment with thioamide drugs is to maintain free T(4) in the upper normal range using the lowest possible dosage. Postpartum thyroiditis requiring thyroxine replacement has been reported in 2% to 5% of women. Most women will return to the euthyroid state within 12 months. PMID- 17077258 TI - Decision-to-incision times and maternal and infant outcomes. PMID- 17077259 TI - A surprising postnatal diagnosis. PMID- 17077260 TI - Decision-to-incision times and maternal and infant outcomes. PMID- 17077265 TI - ACOG Committee Opinion No. 347, November 2006: Using preimplantation embryos for research. AB - Human embryonic stem cell research promises an increased understanding of the molecular process underlying cell differentiation. Transplantation of embryonic stem cells or their derivatives may, in the future, offer therapies for human diseases. In this Committee Opinion, American College of Obstetricians and Gynecologists (ACOG) Committee on Ethics presents an ethical framework for examining issues surrounding research using preimplantation embryos and proposes ethical guidelines for such research. The Committee acknowledges the diversity of opinions among ACOG members and affirms that no physician who finds embryo research morally objectionable should be required or expected to participate in such research. The Committee supports embryo research within 14 days after evidence of fertilization but limits it according to ethical guidelines. The Committee recommends that cryopreserved embryos be the preferred source for research but believes that the promise of somatic cell nuclear transfer is such that research in this area is justified. The Committee opposes reproductive cloning. Intended parents for whom embryos are created should give informed consent for the disposition for any excess embryos. The donors of gametes or somatic cells used in the creation of such tissue should give consent for donation of embryos for research. Potential research projects should be described to potential donors as much as possible. Donation of embryos for stem cell research requires specific consent. The Committee believes that compensation for egg donors for research is acceptable, consistent with American Society for Reproductive Medicine guidelines. PMID- 17077266 TI - ACOG Committee Opinion No. 348, November 2006: Umbilical cord blood gas and acid base analysis. AB - Umbilical cord blood gas and acid-base assessment are the most objective determinations of the fetal metabolic condition at the moment of birth. Moderate and severe newborn encephalopathy, respiratory complications, and composite complication scores increase with an umbilical arterial base deficit of 12-16 mmol/L. Moderate or severe newborn complications occur in 10% of neonates who have this level of acidemia and the rate increases to 40% in neonates who have an umbilical arterial base deficit greater than 16 mmol/L at birth. Immediately after the delivery of the neonate, a segment of umbilical cord should be double clamped, divided, and placed on the delivery table. Physicians should attempt to obtain venous and arterial blood cord samples in circumstances of cesarean delivery for fetal compromise, low 5-minute Apgar score, severe growth restriction, abnormal fetal heart rate tracing, maternal thyroid disease, intrapartum fever, or multifetal gestation. PMID- 17077267 TI - ACOG Committee Opinion No. 349, November 2006: Menstruation in girls and adolescents: using the menstrual cycle as a vital sign. AB - Young patients and their parents often are unsure about what represents normal menstrual patterns, and clinicians also may be unsure about normal ranges for menstrual cycle length and amount and duration of flow through adolescence. It is important to be able to educate young patients and their parents regarding what to expect of a first period and about the range for normal cycle length of subsequent menses. It is equally important for clinicians to have an understanding of bleeding patterns in girls and adolescents, the ability to differentiate between normal and abnormal menstruation, and the skill to know how to evaluate patients' conditions appropriately. Using the menstrual cycle as an additional vital sign adds a powerful tool to the assessment of normal development and the exclusion of serious pathologic conditions. PMID- 17077268 TI - ACOG Committee Opinion No. 350, November 2006: Breast concerns in the adolescent. AB - Breast disease in the adolescent female encompasses an expansive array of topics. Benign disease overwhelmingly dominates the differential diagnosis and dictates a different protocol for care in the adolescent compared with the adult patient to avoid inappropriately high assessments of risk and unnecessary diagnostic procedures and surgery. There also are emerging issues pertaining to the care of the adolescent breast, such as breast augmentation, nipple piercing, and management of the adolescent patient with a family history of breast cancer. PMID- 17077269 TI - ACOG Committee Opinion No. 351, November 2006: The overweight adolescent: prevention, treatment, and obstetric-gynecologic implications. AB - The number of overweight adolescents has grown to epidemic proportions in the United States. Adolescent females who are overweight have significant health sequelae. The American College of Obstetricians and Gynecologists recommends that all adolescents be screened annually for overweight by determining weight and stature, calculating a body mass index for age percentile, and asking about body image and eating patterns. Health care providers should promote healthy eating and physical activity to adolescent patients and their parents during routine preventive health care visits. Adolescents with a body mass index greater than or equal to the 95th percentile for age should have an in-depth dietary and health assessment to determine psychosocial morbidity and risk for future cardiovascular disease. Obstetricians-gynecologists are strongly encouraged to provide this assessment. Additional research is needed to determine the most appropriate approach for the successful prevention and treatment of overweight adolescents. Until this research has been completed, it is best to extrapolate an approach from data and studies pertaining to children and adults, while remaining cognizant of the special needs that surround adolescent growth and development. Sound nutritional recommendations and regular physical activity are essential components of prevention and treatment plans. PMID- 17077270 TI - Stabilization of cellular mRNAs and up-regulation of proteins by oligoribonucleotides homologous to the Bcl2 adenine-uridine rich element motif. AB - Adenine-uridine rich elements (AREs) play an important role in modulating mRNA stability, being the target site of many ARE-binding proteins (AUBPs) that are involved in the decay process. Three 26-mer 2'-O-methyl oligoribonucleotides (ORNs) homologous to the core region of ARE of bcl2 mRNA have been studied for decoy-aptamer activity in UV cross-linking assays. Sense-oriented ORNs competed with the ARE motif for the interaction with both destabilizing and stabilizing AUBPs in cell-free systems and in cell lines. Moreover, ORNs induced mRNA stabilization and up-regulated both Bcl2 mRNA and protein levels in the cells. Bcl2 ORNs stabilized other ARE-containing transcripts and up-regulated their expression. These results indicate that Bcl2 ORNs compete for AUBP-ARE interactions independently of ARE class and suggest that in the cell, the default labile status of ARE-containing mRNAs depends on the combined interaction of such transcripts with destabilizing AUBPs. PMID- 17077271 TI - Roles of a Trypanosoma brucei 5'->3' exoribonuclease homolog in mRNA degradation. AB - The genome of the kinetoplastid parasite Trypanosoma brucei encodes four homologs of the Saccharomyces cerevisiae 5'-->3' exoribonucleases Xrn1p and Xrn2p/Rat1p, XRNA, XRNB, XRNC, and XRND. In S. cerevisiae, Xrn1p is a cytosolic enzyme involved in degradation of mRNA, whereas Xrn2p is involved in RNA processing in the nucleus. Trypanosome XRND was found in the nucleus, XRNB and XRNC were found in the cytoplasm, and XRNA appeared to be in both compartments. XRND and XRNA were essential for parasite growth. Depletion of XRNA increased the abundances of highly unstable developmentally regulated mRNAs, perhaps by delaying a deadenylation-independent decay pathway. Degradation of more stable or unregulated mRNAs was not affected by XRNA depletion although a slight decrease in average poly(A) tail length was observed. We conclude that in trypanosomes 5'- >3' exonuclease activity is important in degradation of highly unstable, regulated mRNAs, but that for other mRNAs another step is more important in determining the decay rate. PMID- 17077272 TI - tmRNA determinants required for facilitating nonstop mRNA decay. AB - In bacteria, ribosomes stalled on nonstop mRNAs are rescued by tmRNA in a unique process called trans-translation. The two known tmRNA functions in trans translation are (1) a tRNA-like function, which transfers the partially synthesized protein fragment to itself; and (2) an mRNA-like function, which enables ribosomes to resume and terminate translation on tmRNA as a surrogate template. We present evidence to demonstrate that tmRNA performs a third function, namely, facilitating the degradation of the causative defective mRNA. Our investigations have revealed the identity of key sequence determinants that promote the degradation of the nonstop mRNA. These sequence determinants are located in the distal part of the tmRNA open reading frame, encoding the ultimate, penultimate, and anti-penultimate amino acids of the peptide tag. We show that mutation of these tmRNA sequence elements has an adverse affect on the disposal of the nonstop mRNA, while leaving the tRNA and mRNA functions entirely unaffected. More significantly, specific mutations that change the nucleotide sequence of the peptide-reading frame without altering the nature or identity of the encoded amino acids still exhibit the characteristic defect in nonstop mRNA decay. In contrast, mutations in codons 3, 4, 5, and 6 of the tmRNA open reading frame do not have an adverse affect on degradation of defective mRNAs. Based on these results, we propose that tmRNA plays an important role in promoting the decay of nonstop mRNAs and that sequence elements in the distal segment of the peptide-reading frame make sequence-specific contributions that are crucial for this activity. PMID- 17077273 TI - Conformational organization of the 3' untranslated region in the tomato bushy stunt virus genome. AB - The 3' untranslated regions (UTRs) of positive-strand RNA viruses often form complex structures that facilitate various viral processes. We have examined the RNA conformation of the 352 nucleotide (nt) long 3' UTR of the Tomato bushy stunt virus (TBSV) genome with the goal of defining both local and global structures that are important for virus viability. Gel mobility analyses of a 3'-terminal 81 nt segment of the 3' UTR revealed that it is able to form a compact RNA domain (or closed conformation) that is stabilized by a previously proposed tertiary interaction. RNA-RNA gel shift assays were used to provide the first physical evidence for the formation of this tertiary interaction and revealed that it represents the dominant or "default" structure in the TBSV genome. Further analysis showed that the tertiary interaction involves five base pairs, each of which contributes differently to overall complex stability. Just upstream from the 3'-terminal domain, a long-distance RNA-RNA interaction involving 3' UTR sequences was found to be required for efficient viral RNA accumulation in vivo and to also contribute to the formation of the 3'-terminal domain in vitro. Collectively, these results provide a comprehensive overview of the conformational and functional organization of the 3' UTR of the TBSV genome. PMID- 17077274 TI - The canonical UPF1-dependent nonsense-mediated mRNA decay is inhibited in transcripts carrying a short open reading frame independent of sequence context. AB - Nonsense-mediated mRNA decay (NMD) is a surveillance mechanism that degrades mRNAs carrying premature translation termination codons. Generally, NMD is elicited if translation terminates >50-54 nucleotides (nt) upstream of an exon exon junction. We have previously reported that human beta-globin mRNAs carrying 5'-proximal nonsense mutations (e.g., beta15) accumulate to normal levels, suggesting an exception to the "50-54-nt boundary rule." In the present report, we demonstrate that the strength of the UPF1-dependent NMD of mutant beta-globin mRNAs is specifically determined by the proximity of the nonsense codon to the initiation AUG. This conclusion is supported by a parallel effect of the short ORF size on NMD of nonsense-containing alpha-globin mRNAs. To determine whether the short-ORF effect on NMD response is conserved in heterologous transcripts, we assessed its effects on a set of beta-globin/triosephosphate isomerase (TPI) hybrid mRNAs and on the TPI mRNA. Our data support the conclusion that nonsense mutations resulting in a short ORF are able to circumvent the full activity of the canonical UPF1-dependent NMD pathway. PMID- 17077275 TI - Gene expression profiles in anatomically and functionally distinct regions of the normal aged human brain. AB - In this article, we have characterized and compared gene expression profiles from laser capture microdissected neurons in six functionally and anatomically distinct regions from clinically and histopathologically normal aged human brains. These regions, which are also known to be differentially vulnerable to the histopathological and metabolic features of Alzheimer's disease (AD), include the entorhinal cortex and hippocampus (limbic and paralimbic areas vulnerable to early neurofibrillary tangle pathology in AD), posterior cingulate cortex (a paralimbic area vulnerable to early metabolic abnormalities in AD), temporal and prefrontal cortex (unimodal and heteromodal sensory association areas vulnerable to early neuritic plaque pathology in AD), and primary visual cortex (a primary sensory area relatively spared in early AD). These neuronal profiles will provide valuable reference information for future studies of the brain, in normal aging, AD and other neurological and psychiatric disorders. PMID- 17077276 TI - Profiling at mRNA, protein, and metabolite levels reveals alterations in renal amino acid handling and glutathione metabolism in kidney tissue of Pept2-/- mice. AB - PEPT2 is an integral membrane protein in the apical membrane of renal epithelial cells that operates as a rheogenic transporter for di- and tripeptides and structurally related drugs. Its prime role is thought to be the reabsorption of filtered di- and tripeptides contributing to amino acid homeostasis. To elucidate the role of PEPT2 in renal amino acid metabolism we submitted kidney tissues of wild-type and a Pept2(-/-) mouse line to a comprehensive transcriptome, proteome and metabolome profiling and analyzed urinary amino acids and dipeptides. cDNA microarray analysis identified 147 differentially expressed transcripts in transporter-deficient animals, and proteome analysis by 2D-PAGE and MALDI-TOF-MS identified 37 differentially expressed proteins. Metabolite profiling by GC-MS revealed predominantly altered concentrations of amino acids and derivatives. Urinary excretion of amino acids demonstrated increased glycine and cysteine/cystine concentrations and dipeptides in urine were assessed by amino acid analysis of urine samples before and after in vitro dipeptidase digestion. Dipeptides constituted a noticeable fraction of urinary amino acids in Pept2(-/-) animals, only, and dipeptide-bound glycine and cystine were selectively increased in Pept2(-/-) urine samples. These findings were confirmed by a drastically increased excretion of cysteinyl-glycine (cys-gly). Urinary loss of cys-gly together with lower concentrations of cysteine, glycine, and oxoproline in kidney tissue and altered expression of mRNA and proteins involved in glutathione (GSH) metabolism suggests that PEPT2 is predominantly a system for reabsorption of cys gly originating from GSH break-down, thus contributing to resynthesis of GSH. PMID- 17077277 TI - Identification of a gene network contributing to hypertrophy in callipyge skeletal muscle. AB - The callipyge mutation in sheep results in postnatal skeletal muscle hypertrophy in the pelvic limbs and loins with little or no effect on anterior skeletal muscles. Associated with the phenotype are changes in the expression of a number of imprinted genes flanking the site of the mutation, which lies in an intergenic region at the telomeric end of ovine chromosome 18. The manner in which these local changes in gene expression are translated into muscle hypertrophy is not known. Microarray-based transcriptional profiling was used to identify differentially expressed genes in longissimus dorsi skeletal muscle samples taken at birth and 12 wk of age from callipyge and wild-type sheep. The phenotype was only expressed at the latter developmental time and associated with decreased type 1 fibers (slow oxidative) and a shift toward type IIx and IIb fibers (fast twitch glycolytic). We have identified 131 genes in the samples taken at 12 wk of age that were differentially expressed as a function of genotype but not due to the fiber type changes. The gene expression changes occurring as a function of genotype in the samples taken at birth indicated that the transcriptional framework underpinning the phenotype was emerging prior to expression of the phenotype. Eight genes were differentially expressed as a function of genotype at both developmental times. A model is proposed describing a core network of genes and histone epigenetic modifications that is likely to underpin the fiber type changes and muscle hypertrophy characteristic of callipyge sheep. PMID- 17077278 TI - Hybrid cardiomyocytes derived by cell fusion in heterotopic cardiac xenografts. AB - Cardiomyocytes expressing host markers, such as the Y chromosome in sex mismatched transplants, have been described in human allografts, suggesting that circulating cells can contribute to cardiac regeneration. It has not been established, however, whether host-derived cardiomyocytes result from transdifferentiation of stem cells or cell fusion. To address this issue, we used heterotopic heart xenografts and looked for markers of donor and recipient cells. Golden Syrian hamsters or transgenic mice expressing nuclear beta-galactosidase under the control of the cardiac troponin I promoter served as organ donors, while GFP+ transgenic rats were used as recipients. GFP+ cells, including abundant CD-45+ inflammatory cells and rare undifferentiated cells expressing early cardiac markers (GATA-4 or MEF2C), were found in xenografts harvested two weeks after surgery. In addition, rare GFP+ mature cardiomyocytes were found in 7 of 8 hamster xenografts and 6 of 6 mouse xenografts. The proportion of these cells was very low (0.0001% to 0.0344% in hamster xenografts) but similar to the one observed in control rat heart allografts. Without exception, all GFP+ cardiomyocytes also expressed donor markers, i.e., hamster membrane antigens or lacZ, so they must derive from cell fusion, not transdifferentiation. PMID- 17077279 TI - Rosiglitazone inhibits mouse liver regeneration. AB - The remarkable regenerative potential of the liver is well known. Recent investigations have shown that this regenerative response is impaired in mouse models of fatty liver disease. Other studies demonstrate that mice engineered for liver-specific overexpression of the peroxisome proliferator activated receptor gamma (PPARgamma) develop significant hepatic steatosis. These observations suggest that precise regulation of hepatic PPARgamma activity may be essential for normal liver regeneration. To test this hypothesis, we analyzed the effects of PPARgamma-activating thiazolidinediones on liver regeneration in the rodent partial hepatectomy model. Thiazolidinediones with different PPARgamma-activating potencies were administered to mice, and those mice were subjected to partial hepatectomy and analyzed for resulting effects on hepatocellular proliferation and signaling pathways important during normal liver regeneration. The results showed that thiazolidinediones suppress liver regeneration with efficacies that correlate with their relative PPARgamma-activating potencies. These studies provide the first evidence linking regulation of PPARgamma activity and the hepatic regenerative response. PMID- 17077280 TI - Evidence of post-transcriptional regulation in the maintenance of a partial muscle phenotype by electrogenic cells of S. macrurus. AB - Electrocytes, the current-producing cells of electric organs (EOs) in electric fish, are unique in that they derive from striated muscle and they possess biochemical characteristics of both muscle and non-muscle cells. In the freshwater teleost Sternopygus macrurus, electrocytes are multinucleated cells that do not contract yet retain expression of some proteins common to skeletal muscle cells. Given the role that transcriptional regulation plays in the activation of the myogenic program in vertebrates, we examined the expression patterns of several genes associated with multiple functions of skeletal muscle in mature electrocytes of S. macrurus. Our expression analyses detected transcripts for alpha-actin, alpha-acetylcholine (ACh) receptor (alpha-AChR), desmin, muscle creatine kinase (MCK), myosin heavy chain (MHC) isoforms, titin, tropomyosin, and troponin-T genes in the EO. However, immunolabeling studies revealed that electrocytes do not contain MCK, MHCs, or tropomyosin or troponin-T proteins. These results underscore the contribution of gene regulatory mechanisms in the maintenance of the muscle-like phenotype of EO that may be transcriptional independent. We also report the classification and frequency of distinct transcripts from a random selection of 420 clones from an EO cDNA library. This is the first characterization of expressed genes in an EO, and it is an important step toward identifying mechanisms that affect different muscle protein systems for the evolution of highly specialized noncontractile tissues. Evidence of post transcriptional regulation in the maintenance of a partial muscle phenotype by electrogenic cells of S. macrurus. PMID- 17077281 TI - Regulation of hepatic cholesterol synthesis by a novel protein (SPF) that accelerates cholesterol biosynthesis. AB - Supernatant protein factor (SPF) is a novel cholesterol biosynthesis-accelerating protein expressed in liver and small intestine. Here, we report on the physiological role of SPF by using Spf-deficient mice. Although plasma cholesterol levels were similar in chow-fed Spf-/- and wild-type (WT) mice, fasting significantly decreased plasma cholesterol levels in Spf-/- mice but not in WT mice. While fasting reduced hepatic cholesterol synthesis rate in WT mice, a more pronounced reduction was observed in Spf-/- mice. The expression of cholesterogenic enzymes was dramatically suppressed by fasting both in WT and Spf /- mice. In contrast, hepatic SPF expression of WT mice was up-regulated by fasting in peroxisome proliferator-activated receptor alpha (PPAR-alpha) dependent manner. These results indicate that in WT mice, the decrease of hepatic cholesterol synthesis under fasting conditions is at least in part compensated by SPF up-regulation. Fibrates, which function as a PPAR-alpha agonist and are widely used as hypotriglycemic drugs, reduced hepatic cholesterol synthesis and plasma cholesterol levels by approximately one-half in Spf-/- mice but not in WT mice. These findings suggest that co-administration of fibrates and an SPF inhibitor may reduce not only plasma triglyceride but also cholesterol levels, indicating that SPF is a promising hypocholesterolemic drug target. PMID- 17077282 TI - The calcium sensing receptor is directly involved in both osteoclast differentiation and apoptosis. AB - Intracellular transduction pathways that are dependent on activation of the CaR by Ca(o)2+ have been studied extensively in parathyroid and other cell types, and include cytosolic calcium, phospholipases C, A2, and D, protein kinase C isoforms and the cAMP/protein kinase A system. In this study, using bone marrow cells isolated from CaR-/- mice as well as DN-CaR-transfected RAW 264.7 cells, we provide evidence that expression of the CaR plays an important role in osteoclast differentiation. We also establish that activation of the CaR and resultant stimulation of PLC are involved in high Ca(o)2+-induced apoptosis of mature rabbit osteoclasts. Similar to RANKL, Ca(o)2+ (20 mM) appeared to trigger rapid and significant nuclear translocation of NF-kappaB in a CaR- and PLC-dependent manner. In summary, our data suggest that stimulation of the CaR may play a pivotal role in the control of both osteoclast differentiation and apoptosis in the systems studied here through a signaling pathway involving activation of the CaR, phospholipase C, and NF-kappaB. PMID- 17077283 TI - Platelets induce differentiation of human CD34+ progenitor cells into foam cells and endothelial cells. AB - Recruitment of human CD34+ progenitor cells toward vascular lesions and differentiation into vascular cells has been regarded as a critical initial step in atherosclerosis. Previously we found that adherent platelets represent potential mediators of progenitor cell homing besides their role in thrombus formation. On the other hand, foam cell formation represents a key process in atherosclerotic plaque formation. To investigate whether platelets are involved in progenitor cell recruitment and differentiation into endothelial cells and foam cells, we examined the interactions of platelets and CD34+ progenitor cells. Cocultivation experiments showed that human platelets recruit CD34+ progenitor cells via the specific adhesion receptors P-selectin/PSGL-1 and beta1- and beta2 integrins. Furthermore, platelets were found to induce differentiation of CD34+ progenitor cells into mature foam cells and endothelial cells. Platelet-induced foam cell generation could be prevented partially by HMG coenzyme A reductase inhibitors via reduction of matrix metalloproteinase-9 (MMP-9) secretion. Finally, agonists of peroxisome proliferator-activated receptor-alpha and -gamma attenuated platelet-induced foam cell generation and production of MMP-9. The present study describes a potentially important mechanism of platelet-induced foam cell formation and generation of endothelium in atherogenesis and atheroprogression. The understanding and modulation of these mechanisms may offer new treatment strategies for patients at high risk for atherosclerotic diseases. PMID- 17077284 TI - 3'Phosphoinositide-dependent kinase-1 is essential for ischemic preconditioning of the myocardium. AB - Brief periods of ischemia and reperfusion that precede sustained ischemia lead to a reduction in myocardial infarct size. This phenomenon, known as ischemic preconditioning, is mediated by signaling pathway(s) that are yet to be fully defined. 3'-Phosphoinositide-dependent kinase-1 (PDK1) has been implicated in numerous cellular processes. However, the involvement of PDK1 in preconditioning has yet to be elucidated. Studying PDK1 is not as straightforward as it is for the majority of kinases, due to the lack of a specific inhibitor of PDK1. Therefore, we have taken advantage of PDK1 hypomorphic mutant mice with reduced expression of PDK1 to study the role of PDK1 in preconditioning. Whole heart and single cell models of preconditioning demonstrated that the hearts and cardiac cells from PDK1 hypomorphic mice could not be preconditioned. The cardioprotective effect of PDK1 was not related to the effect that preconditioning has on sarcolemmal membrane action potential as revealed by di-8 ANEPPS, a sarcolemmal-potential sensitive dye, and laser confocal microscopy. In contrast, experiments with JC-1, a mitochondrial membrane potential-sensitive dye, has demonstrated that intact PDK1 levels were required for preconditioning mediated regulation of mitochondrial membrane potential. Western blotting combined with functional experiments have shown that intact PDK1 levels were required for preconditioning-induced phosphorylation of protein kinase B (PKB), glycogen synthase kinase-3beta (GSK-3beta), and cardioprotection. We conclude that PDK1 mediates preconditioning in the heart by regulating activating PKB-GSK 3beta to regulate mitochondrial but not sarcolemmal membrane potential. 3'Phosphoinositide-dependent kinase-1 (PDK1) is essential for ischemic preconditioning of the myocardium. PMID- 17077285 TI - Glucocorticoid resistance in two key models of acute lymphoblastic leukemia occurs at the level of the glucocorticoid receptor. AB - Glucocorticoids (GCs) specifically induce apoptosis in malignant lymphoblasts and are thus pivotal in the treatment of acute lymphoblastic leukemia (ALL). However, GC-resistance is a therapeutic problem with an unclear molecular mechanism. We generated approximately 70 GC-resistant sublines from a GC-sensitive B- and a T ALL cell line and investigated their mechanisms of resistance. In response to GCs, all GC-resistant subclones analyzed by real-time polymerase chain reaction (PCR) showed a deficient up-regulation of the GC-receptor (GR) and its downstream target, GC-induced leucine zipper. This deficiency in GR up-regulation was confirmed by Western blotting and on retroviral overexpression of GR in resistant subclones GC-sensitivity was restored. All GC-resistant subclones were screened for GR mutations using denaturing high-pressure liquid chromatography (DHPLC), DNA-fingerprinting, and fluorescence in situ hybridization (FISH). Among the identified mutations were some previously not associated with GC resistance: A484D, P515H, L756N, Y663H, L680P, and R714W. This approach revealed three genotypes, complete loss of functional GR in the mismatch repair deficient T-ALL model, apparently normal GR genes in B-ALLs, and heterozygosity in both. In the first genotype, deficiency in GR up-regulation was fully explained by mutational events, in the second by a putative regulatory defect, and in the third by a combination thereof. In all instances, GC-resistance occurred at the level of the GR in both models. PMID- 17077286 TI - A novel formulation of oxygen-carrying matrix enhances liver-specific function of cultured hepatocytes. AB - Oxygen is an important component of the cellular microenvironment, mediating cell survival, differentiation, and function. Oxygen supply is a limiting factor during culture of highly metabolic cells such as hepatocytes. Here we present a simple formulation of a fluorocarbon-based oxygen carrier embedded in collagen gel that increases oxygen concentration in culture 6-fold. Rat hepatocytes cultured on oxygen carrier-collagen showed a significant increase in viability and function. Cytochrome P450IA1 activity was increased by 140% in serum-free cultures and by 820% in serum-containing cultures. The significantly higher hepatocellular function on oxygen carrier-collagen matrix persisted and increased during long-term culture. Long-term albumin secretion was increased by 350% in serum-free cultures and by 166% in serum-containing culture. Long-term urea secretion was increased by 79% in serum-free cultures and by 76% in serum containing cultures. We conclude that oxygen supply may limit hepatocyte function in vitro. This limitation can be overcome by addition of an oxygen carrier to the extracellular matrix. Culture of hepatocytes on oxygen-carrying matrix mimics the oxygen-rich environment of the liver and provides a simple method for enhanced long-term function. PMID- 17077287 TI - Acquisition of in vitro and in vivo functionality of Nurr1-induced dopamine neurons. AB - Neural precursor cells provide an expandable source of neurons and glia for basic and translational applications. However, little progress has been made in directing naive neural precursors toward specific neuronal fates such as midbrain dopamine (DA) neurons. We have recently demonstrated that transgenic expression of the nuclear orphan receptor Nurr1 is sufficient to drive dopaminergic differentiation of forebrain embryonic rat neural precursors in vitro. However, Nurr1-induced DA neurons exhibit immature neuronal morphologies and functional properties and are unable to induce behavioral recovery in rodent models of Parkinson's disease (PD). Here, we report on the identification of key genetic factors that drive morphological and functional differentiation of Nurr1-derived DA neurons. We show that coexpression of Nurr1, Bcl-XL, and Sonic hedgehog (SHH) or Nurr1 and the proneural bHLH factor Mash1 is sufficient to drive naive rat forebrain precursors into neurons exhibiting the biochemical, electrophysiological, and functional properties of DA neuron in vitro. On transplantation into the striatum of Parkinsonian rats, precursor cells engineered with Nurr1/SHH/Bcl-XL or Nurr1/Mash1 survived in vivo and differentiated into mature DA neurons that can reverse the behavioral deficits in the grafted animals. PMID- 17077288 TI - An invertebrate-like phototransduction cascade mediates light detection in the chicken retinal ganglion cells. AB - Prebilaterian animals perceived ambient light through nonvisual rhabdomeric photoreceptors (RPs), which evolved as support of the chordate visual system. In vertebrates, the identity of nonvisual photoreceptors and the phototransduction cascade involved in nonimage forming tasks remain uncertain. We investigated whether chicken retinal ganglion cells (RGCs) could be nonvisual photoreceptors and the nature of the photocascade involved. We found that primary cultures of chicken embryonic RGCs express such RP markers as transcription factors Pax6 and Brn3, photopigment melanopsin, and G-protein q but not markers for ciliary photoreceptors (alpha-transducin and Crx). To investigate the photoreceptive capability of RGCs, we assessed the direct effect of light on 3H-melatonin synthesis in RGC cultures synchronized to 12:12 h light-dark cycles. In constant dark, RGCs displayed a daily variation in 3H-melatonin levels peaking at subjective day, which was significantly inhibited by light. This light effect was further increased by the chromophore all-trans-retinal and suppressed by specific inhibitors of the invertebrate photocascade involving phosphoinositide hydrolysis (100 microM neomycin; 5 microM U73122) and Ca2+ mobilization (10 mM BAPTA; 1 mM lanthanum). The results demonstrate that chicken RGCs are intrinsically photosensitive RPs operating via an invertebrate-like phototransduction cascade, which may be responsible for early detection of light before vision occurs. PMID- 17077289 TI - Atherogenic properties of LDL particles modified by human group X secreted phospholipase A2 on human endothelial cell function. AB - Increasing evidence suggests that secreted phospholipases A2 (sPLA2s) play an important role in the pathophysiology of atherosclerosis. Among sPLA2s, the human group X (hGX) enzyme has the highest catalytic activity toward phosphatidylcholine, one of the major phospholipid species of cell membranes and low-density lipoprotein (LDL). Our study examined the presence of hGX sPLA2 in human atherosclerotic lesions and investigated the ability of hGX modified LDL to alter human endothelial cell (HUVEC) function. Our results show that hGX sPLA2 is present in human atherosclerotic lesions and that the hydrolysis of LDL by hGX sPLA2 results in a modified particle that induces lipid accumulation in human monocyte-derived macrophages. Acting on endothelial cells, hGX-modified LDL activates the MAP kinase pathway, which leads to increased arachidonic acid release, increased expression of adhesion molecules on the surface of HUVEC, and increased adhesion of monocytes to HUVEC monolayers. Together, our data suggest that LDL modified by hGX, rather than hGX itself may have strong proinflammatory and proatherogenic properties, which could play an important role in the propagation of atherosclerosis. PMID- 17077290 TI - Leptin signaling in neurotensin neurons involves STAT, MAP kinases ERK1/2, and p38 through c-Fos and ATF1. AB - The adipokine leptin signals energy status to the hypothalamus, which triggers a network of neuropeptide responses. Each hypothalamic cell type expresses a unique complement of neuropeptides, receptors, and second messengers; thus each likely responds specifically to peripheral hormones. We describe here the analysis of leptin signaling in a clonal population of mouse neurotensin (NT) -expressing hypothalamic neurons, N-39. Leptin induced phosphorylation of STAT3 and MAPK ERK1/2, but not the downstream effector of PI3K, Akt, and also induced c-Fos protein. We found activation of p38 MAPK by leptin, accompanied by phosphorylation of its downstream effector ATF-1. Phosphorylation of ATF-1 is blocked by the p38 MAPK inhibitor SB 203580. We linked this signaling directly to NT transcription. Protein binding analysis indicates that both ATF-1 and c-Fos are capable of binding to the mouse NT/N gene predominantly at physiological or high concentrations of leptin. The evidence indicates activation of distinct leptin signal transduction pathways that directly result in changes in NT gene expression and links these specific neurons to the control of energy homeostasis. PMID- 17077291 TI - The extracellular adherence protein from Staphylococcus aureus abrogates angiogenic responses of endothelial cells by blocking Ras activation. AB - The extracellular adherence protein (Eap), a broad-spectrum adhesin secreted by Staphylococcus aureus, was previously shown to curb acute inflammatory responses, presumably through its binding to endothelial cell (EC) ICAM-1. Examining the effect of Eap on endothelial function in more detail, we here show that, in addition, Eap functions as a potent angiostatic agent. Concomitant treatment of EC with purified Eap resulted in the complete blockage of the mitogenic and sprouting responses elicited by vascular endothelial growth factor (VEGF)165 or basic fibroblast growth factor (bFGF). Moreover, the induction of tissue factor and decay-accelerating factor were repressed by Eap, as determined by qRT polymerase chain reaction (qRT-PCR), with a corresponding reduction in Egr-1 protein up-regulation seen. This angiostatic activity was accompanied by a corresponding inhibition in ERK1/2 phosphorylation, while activation of p38 was not affected. Inhibition occurred downstream of tyrosine kinase receptor activation, as comparable effects were seen on TPA-induced ERK1/2 phosphorylation. Similar to previously described angiostatic agents like angiopoietin-1 or the 16-kDa prolactin fragment, Eap blockage of the Ras/Raf/MEK/ERK cascade was localized by pull-down assay at the level of Ras activation. Eap's combined anti-inflammatory and antiangiogenic properties render this bacterial protein not only an important virulence factor during S. aureus infection but open new perspectives for therapeutic applications in pathological neovascularization. PMID- 17077293 TI - Teach evolution, learn science: we're ahead of Turkey, but behind Iran. PMID- 17077294 TI - Reflections on the prize of prizes: Alfred Nobel. PMID- 17077295 TI - A globin for the brain. AB - The discovery that a myoglobin-like hemeprotein (called neuroglobin) is expressed in our brain raised considerable curiosity from the standpoints of biochemistry and pathophysiology alike. Neuroglobin is involved in neuroprotection from damage due to hypoxia or ischemia in vitro and in vivo; overexpression of neuroglobin ameliorates the recovery from stroke in experimental animals. The mechanism underlying this remarkable effect is still mysterious. Structural studies revealed that neuroglobin has a typical globin fold, and despite being hexacoordinated, it binds reversibly O2, CO, and NO, undergoing a substantial conformational change of the heme and of the protein. The possible mechanisms involved in neuroprotection are briefly reviewed. Neuroglobin is unlikely to be involved in O2 transport (like myoglobin), although it seems to act as a sensor of the O2/NO ratio in the cell, possibly regulating the GDP/GTP exchange rate forming a specific complex with the G(alpha beta gamma)-protein when oxidized but not when bound to a gaseous ligand. Thus it appears that neuroglobin is a stress responsive sensor for signal transduction in the brain, mediated by a ligand linked conformational change of the protein. PMID- 17077296 TI - The immunoregulatory effects of HIV-1 Nef on dendritic cells and the pathogenesis of AIDS. AB - Dendritic cells (DC) play a crucial role in the generation and regulation of immunity, and their interaction with HIV is relevant in the pathogenesis of AIDS favoring both the initial establishment and spread of the infection and the development of antiviral immunity. HIV-1 Nef is an essential factor for efficient viral replication and pathogenesis, and several studies have been addressed to assess the possible influence of endogenous or exogenous Nef on DC biology. Our findings and other reported data described in this review demonstrate that Nef subverts DC biology interfering with phenotypical, morphological, and functional DC developmental programs, thus representing a viral tool underlying AIDS pathogenesis. This review provides an overview on the mechanism by which Nef, hijacking DC functional activity, may favor both the replication of HIV-1 and the escape from immune surveillance. Overall, the findings described here may contribute to the understanding of Nef function, mechanism of action, and cellular partners. Further elucidation of genes induced through Nef signaling in DC could reveal pathways used by DC to drive HIV spread and will be critical to identify therapeutic strategies to bias the DC system toward activation of antiviral immunity instead of facilitating virus dissemination. PMID- 17077297 TI - Interruption of cell transformation and cancer formation. AB - A review of the results of X-ray and chemical carcinogen induction of transformation of mouse cells supports a two-step epigenetic model of transformation. According to this model, exposure induces an epigenetic regulatory alteration that makes the cells hypermutable so that when the cell population inheriting this alteration becomes sufficiently large, the second step, a mutation to the transformant phenotype, becomes increasingly likely. The epigenetic alteration in X-ray-exposed mouse cells has been demonstrated to be reversible by brief exposure to certain protease inhibitors. If the rodent cell experiments constitute a valid system for studying human cancer, then this two step model may herald rich opportunities for preventing and perhaps even treating cancer in humans. PMID- 17077298 TI - Reduced intestinal and renal amino acid transport in PDK1 hypomorphic mice. AB - The phosphoinositide-dependent kinase PDK1 activates the serum- and glucocorticoid-inducible kinase isoforms SGK1, SGK2, and SGK3 and protein kinase B, which in turn are known to up-regulate a variety of sodium-coupled transporters. The present study was performed to explore the role of PDK1 in amino acid transport. As mice completely lacking functional PDK1 are not viable, mice expressing 10-25% of PDK1 (pdk1(hm)) were compared with their wild-type (WT) littermates (pdk1(wt)). Body weight was significantly less in pdk1(hm) than in pdk1(wt) mice. Despite lower body weight of pdk1(hm) mice, food and water intake were similar in pdk1(hm) and pdk1(wt) mice. According to Ussing chamber experiments, electrogenic transport of phenylalanine, cysteine, glutamine, proline, leucine, and tryptophan was significantly smaller in jejunum of pdk1(hm) mice than in pdk1(wt) mice. Similarly, electrogenic transport of phenylalanine, glutamine, and proline was significantly decreased in isolated perfused proximal tubules of pdk1(hm) mice. The urinary excretion of proline, valine, guanidinoacetate, methionine, phenylalanine, citrulline, glutamine/glutamate, and tryptophan was significantly larger in pdk1(hm) than in pdk1(wt) mice. According to immunoblotting of brush border membrane proteins prepared from kidney, expression of the Na+-dependent neutral amino acid transporter B(0)AT1 (SLC6A19), the glutamate transporter EAAC1/EAAT3 (SLC1A1), and the transporter for cationic amino acids and cystine b(0,+)AT (SLC7A9) was decreased but the Na+/proline cotransporter SIT (SLC6A20) was increased in pdk1(hm) mice. In conclusion, reduction of functional PDK1 leads to impairment of intestinal absorption and renal reabsorption of amino acids. The combined intestinal and renal loss of amino acids may contribute to the growth defect of PDK1-deficient mice. PMID- 17077299 TI - The dopamine D3 receptor plays an essential role in alcohol-seeking and relapse. AB - Our study aimed to identify new candidate genes, which might be involved in alcohol craving and relapse. To find changes in gene expression after long-term alcohol consumption, we studied gene expression profiles in the striatal dopamine system by using DNA microarrays of two different alcohol-preferring rat lines (HAD and P). Our data revealed an up-regulation of the dopamine D3 receptor (D3R) after 1 yr of voluntary alcohol consumption in the striatum of alcohol preferring rats that was confirmed by qRT-polymerase chain reaction. This finding was further supported by the finding of up-regulated striatal D3R mRNA in nonselected Wistar rats after long-term alcohol consumption when compared with age-matched control animals. We further examined the role of the D3R in mediating alcohol relapse behavior using the alcohol deprivation effect (ADE) model in long-term alcohol drinking Wistar rats and the model of cue-induced reinstatement of alcohol-seeking behavior using the selective D3R antagonist SB-277011-A (0, 1, 3, and 10 mg/kg) and the partial agonist BP 897 (0, 0.1, 1, and 3 mg/kg). Both treatments caused a dose-dependent reduction of relapse-like drinking in the ADE model as well as a decrease in cue-induced ethanol-seeking behavior. We conclude that long-term alcohol consumption leads to an up-regulation of the dopamine D3R that may contribute to alcohol-seeking and relapse. We therefore suggest that selective antagonists of this pharmacological target provide a specific treatment approach to reduce alcohol craving and relapse behavior. PMID- 17077300 TI - Melanocortin 3 receptors control crystal-induced inflammation. AB - In this study we have characterized the anti-inflammatory profile of a selective melanocortin type 3 receptor (MC3-R) ligand [D-Trp8]-gamma-MSH, validating in vitro results with analyses in mice deficient for this receptor subtype. In wild type (WT) macrophages, [D-Trp8]-gamma-MSH activated MC3-R (as tested by accumulation of cyclic AMP) and inhibited (approximately 50%) the release of interleukin (IL)-1 and the chemokine KC (CXCL1), but was ineffective in cells taken from MC3-R null mice. In vivo, administration of 3-30 microg [D-Trp8]-gamma MSH significantly inhibited leukocyte influx and cytokine production in a model of crystal-induced peritonitis, and these effects were absent in MC3-R null mice or blocked by coadministration of an MC3-R antagonist. Finally, in a model of gouty arthritis, direct injection of urate crystals into the rat joint provoked a marked inflammatory reaction that was significantly inhibited (approximately 70%) by systemic or local administration of [D-Trp8]-gamma-MSH. In conclusion, using an integrated transgenic and pharmacological approach, we provide strong proof of concept for the development of selective MC3-R agonists as novel anti inflammatory therapeutics. PMID- 17077301 TI - HIF-dependent induction of adenosine A2B receptor in hypoxia. AB - Adenosine has been widely associated with hypoxia of many origins, including those associated with inflammation and tumorogenesis. A number of recent studies have implicated metabolic control of adenosine generation at sites of tissue hypoxia. Here, we examine adenosine receptor control and amplification of signaling through transcriptional regulation of endothelial and epithelial adenosine receptors. Initial studies confirmed previous findings indicating selective induction of human adenosine A2B receptor (A2BR) by hypoxia. Analysis of the cloned human A2BR promoter identified a functional hypoxia-responsive region, including a functional binding site for hypoxia-inducible factor (HIF) within the A2BR promoter. Further studies examining HIF-1alpha DNA binding and HIF-1alpha gain and loss of function confirmed strong dependence of A2BR induction by HIF-1alpha in vitro and in vivo mouse models. Additional studies in endothelia overexpressing full-length A2BR revealed functional phenotypes of increased barrier function and enhanced angiogenesis. Taken together, these results demonstrate transcriptional coordination of A2BR by HIF-1alpha and amplified adenosine signaling during hypoxia. These findings may provide an important link between hypoxia and metabolic conditions associated with inflammation and angiogenesis. PMID- 17077302 TI - Production of germline transgenic chickens expressing enhanced green fluorescent protein using a MoMLV-based retrovirus vector. AB - The Moloney murine leukemia virus (MoMLV) -based retrovirus vector system has been used most often in gene transfer work, but has been known to cause silencing of the imported gene in transgenic animals. In the present study, using a MoMLV based retrovirus vector, we successfully generated a new transgenic chicken line expressing high levels of enhanced green fluorescent protein (eGFP). The level of eGFP expression was conserved after germline transmission and as much as 100 microg of eGFP could be detected per 1 mg of tissue protein. DNA sequencing showed that the transgene had been integrated at chromosome 26 of the G1 and G2 generation transgenic chickens. Owing to the stable integration of the transgene, it is now feasible to produce G3 generation of homozygous eGFP transgenic chickens that will provide 100% transgenic eggs. These results will help establish a useful transgenic chicken model system for studies of embryonic development and for efficient production of transgenic chickens as bioreactors. PMID- 17077303 TI - Human ACE and bradykinin B2 receptors form a complex at the plasma membrane. AB - To investigate how angiotensin I-converting enzyme (ACE) inhibitors enhance the actions of bradykinin (BK) on B2 receptors independent of blocking BK inactivation, we expressed human somatic ACE and B2 receptors in CHO cells. Bradykinin and its ACE-resistant analog were the receptor agonists. B2 fused with green fluorescent protein (GFP) and ACE were coprecipitated with antisera to GFP or ACE shown in Western blots. Immunohistochemistry of fixed cells localized ACE by red color and B2-GFP by green. Yellow on plasma membranes of coexpressing cells also indicated enzyme-receptor complex formation. Using ACE-fused cyan fluorescent protein donor and B2-fused yellow fluorescent protein (YFP) acceptor, we registered fluorescence resonance energy transfer (FRET) by the enhanced fluorescence of donor on acceptor photobleaching, establishing close (within 10 nm) positions of B2 receptors and ACE. Bradykinin stimulation cointernalized ACE and B2 receptors. We expressed ACE fused to N terminus of B2 receptors, anchoring only receptors to plasma membranes. Here, in contrast to cells, where both ACE and B2 receptors are separately anchored, ACE inhibitors neither enhance activation of chimeric B2 nor resensitize desensitized B2 receptors. Heterodimer formation between ACE and B2 receptors can be a mechanism for ACE inhibitors to augment kinin activity at cellular level. PMID- 17077304 TI - Gene knockout of the KCNJ8-encoded Kir6.1 K(ATP) channel imparts fatal susceptibility to endotoxemia. AB - Sepsis, the systemic inflammatory response to infection, imposes a high demand for bodily adaptation, with the cardiovascular response a key determinant of outcome. The homeostatic elements that secure cardiac tolerance in the setting of the sepsis syndrome are poorly understood. Here, in a model of acute septic shock induced by endotoxin challenge with Escherichia coli lipopolysaccharide (LPS), knockout of the KCNJ8 gene encoding the vascular Kir6.1 K(ATP) channel pore predisposed to an early and profound survival disadvantage. The exaggerated susceptibility provoked by disruption of this stress-responsive sensor of cellular metabolism was linked to progressive deterioration in cardiac activity, ischemic myocardial damage, and contractile dysfunction. Deletion of KCNJ8 blunted the responsiveness of coronary vessels to cytokine- or metabolic-mediated vasodilation necessary to support myocardial perfusion in the wild-type (WT), creating a deficit in adaptive response in the Kir6.1 knockout. Application of a K(ATP) channel opener drug improved survival in the endotoxic WT but had no effect in the Kir6.1 knockout. Restoration of the dilatory capacity of coronary vessels was required to rescue the Kir6.1 knockout phenotype and reverse survival disadvantage in lethal endotoxemia. Thus, the Kir6.1-containing K(ATP) channel, by coupling vasoreactivity with metabolic demand, provides a vital feedback element for cardiovascular tolerance in endotoxic shock. PMID- 17077305 TI - Cytocidal effects of atheromatous plaque components: the death zone revisited. AB - OBJECTIVE: Earlier we suggested that atheroma lesions constitute a "death zone" containing toxic materials that may cause dysfunction and demise of invading macrophages to prevent the removal of plaque materials. Here we have assessed the cytotoxic effects of nonfractionated gruel and insoluble (ceroid-like) material derived from advanced human atheroma. METHODS AND RESULTS: The insoluble material within advanced atherosclerotic plaque was isolated following protease K digestion and extensive extraction with aqueous and organic solvents. FTIR, Raman, and atomic absorption spectroscopy suggested that, despite its fluorescent nature, this material closely resembled hydroxyapatite and dentin, but also contained a significant amount of iron and calcium. When added to J774 cells and human macrophages in culture, this insoluble substance was phagocytosed, and progressive cell death followed. However, an even more cytotoxic activity was found in the atheromatous "gruel" that contains abundant carbonyls/aldehydes. Cell death caused by both crude gruel and ceroid could be blocked by preincubating cells with the lipophilic iron chelator salicylaldehyde isonicotinoyl hydrazone, apoferritin, BAPTA/AM, or sodium borohydride, indicating that cellular iron, calcium, and reactive aldehyde(s) are responsible for the observed cytotoxicity. CONCLUSIONS: Toxic materials within atheromatous lesions include both ceroid and even more cytotoxic lipidaceous materials. The cytotoxic effects of these plaque components may help explain the persistence of atherosclerotic lesions. PMID- 17077306 TI - Expression of T-cadherin in tumor cells influences invasive potential of human hepatocellular carcinoma. AB - Overexpression of T-cadherin (T-cad) transcripts occurs in approximately 50% of human hepatocellular carcinomas (HCCs). To elucidate T-cad functions in HCC, we examined T-cad protein expression in normal and tumoral human livers and hepatoma cell lines and investigated its influence on invasive potential of HCC using RNA interference silencing of T-cad expression in Mahlavu cells. Whereas T-cad expression was restricted to endothelial cells (EC) from large blood vessels in normal livers, it was up-regulated in sinusoidal EC from 8/15 invasive HCCs. Importantly, in three of them (38%) T-cad was detected in tumor cells within regions in which E-cadherin expression was absent. Among six hepatoma cell lines, only Mahlavu expressed T-cad but not E-cadherin. T-cad exhibited a globally punctuate distribution in quiescent Mahlavu and additionally it concentrated at the leading edge of migrating cells. Matrigel invasion assay revealed that Mahlavu possess a high invasive potential that was significantly inhibited by T cad silencing. Wound healing and random motility assays demonstrated that inhibition of T-cad expression in Mahlavu significantly reduced their motility. We propose that T-cad expression in tumor cells might occur by cadherin-switching during epithelial-mesenchymal transition and may represent an additional mechanism contributing to HCC metastasis. PMID- 17077307 TI - Alpha-synuclein induces hyperphosphorylation of Tau in the MPTP model of parkinsonism. AB - Many neurodegenerative diseases associated with functional Tau dysregulation, including Alzheimer's disease (AD) and other tauopathies, also show alpha synuclein (alpha-Syn) pathology, a protein associated with Parkinson's disease (PD) pathology. Here we show that treatment of primary mesencephalic neurons (48 h) or subchronic treatment of wild-type (WT) mice with the Parkinsonism-inducing neurotoxin MPP+/MPTP, results in selective dose-dependent hyperphosphorylation of Tau at Ser396/404 (PHF-1-reactive Tau, p-Tau), with no changes in pSer202 but with nonspecific increases in pSer262 levels. The presence of alpha-Syn was absolutely mandatory to observe MPP+/MPTP-induced increases in p-Tau levels, since no alterations in p-Tau were seen in transfected cells not expressing alpha Syn or in alpha-Syn-/- mice. MPP+/MPTP also induced a significant accumulation of alpha-Syn in both mesencephalic neurons and in WT mice striatum. MPTP/MPP+ lead to differential alterations in p-Tau and alpha-Syn levels in a cytoskeleton bound, vs. a soluble, cytoskeleton-free fraction, inducing their coimmunoprecipitation in the cytoskeleton-free fraction and neuronal soma. Subchronic MPTP exposure increased sarkosyl-insoluble p-Tau in striatum of WT but not alpha-Syn-/- mice. These studies describe a novel mechanism for MPTP neurotoxicity, namely a MPTP-inducible, strictly alpha-Syn-dependent, increased formation of PHF-1-reactive Tau, suggesting convergent overlapping pathways in the genesis of clinically divergent diseases such as AD and PD. PMID- 17077308 TI - Moderate consumption of Cabernet Sauvignon attenuates Abeta neuropathology in a mouse model of Alzheimer's disease. AB - Recent studies suggest that moderate red wine consumption reduces the incidence of Alzheimer's disease (AD) clinical dementia. Using Tg2576 mice, which model AD type amyloid beta-protein (Abeta) neuropathology, we tested whether moderate consumption of the red wine Cabernet Sauvignon modulates AD-type neuropathology and cognitive deterioration. The wine used in the study was generated using Cabernet Sauvignon grapes from Fresno, California, and was delivered to Tg2576 in a final concentration of approximately 6% ethanol. We found that Cabernet Sauvignon significantly attenuated AD-type deterioration of spatial memory function and Abeta neuropathology in Tg2576 mice relative to control Tg2576 mice that were treated with either a comparable amount of ethanol or water alone. Chemical analysis showed the Cabernet Sauvignon used in this study contains a very low content of resveratrol (0.2 mg/L), 10-fold lower than the minimal effective concentration shown to promote Abeta clearance in vitro. Our studies suggest Cabernet Sauvignon exerts a beneficial effect by promoting nonamyloidogenic processing of amyloid precursor protein, which ultimately prevents the generation of Abeta peptides. This study supports epidemiological evidence indicating that moderate wine consumption, within the range recommended by the FDA dietary guidelines of one drink per day for women and two for men, may help reduce the relative risk for AD clinical dementia. PMID- 17077309 TI - The spleen as a target in severe acute respiratory syndrome. AB - It has been proposed that immune injury is the central mechanism of pathogenesis of the infectious disease, severe acute respiratory syndrome (SARS). To gain a better understanding of immune injury in the spleen, we investigated the number and distribution of various immune cell types in the spleens of SARS patients. We performed autopsies on six confirmed SARS cases, with six normal subjects as controls; spleen samples from these autopsies were examined with hematoxylin and eosin (H&E) sections, in situ hybridization for SARS virus genomic sequences, and immunohistochemistry with seven monoclonal antibodies to five cell types. The number and distribution of these cells were measured and analyzed using an image analysis system. SARS genomic sequences were detected in all SARS spleens. The SARS spleens all had severe damage to the white pulp and showed an alteration of the normal distribution of various cell types. Immunocytes in the red pulp were decreased by 68.0-90.7% except for CD68+ macrophages and human leukocyte antigen (HLA)-DR positive antigen-presenting cells (APC), which were decreased to a lesser degree. On average, CD68+ macrophages were increased in size by 2.21-fold. We hypothesize that the collapse of the splenic immune system plays a key role in the clinical outcome of these patients. PMID- 17077310 TI - Species specificity of mammalian connexin-26 to form open voltage-gated hemichannels. AB - Mutations of connexin-26 (Cx26) cause nonsyndromic hearing loss and other syndromes affecting ectoderm-derived tissues. While the exact mechanisms underlying these diseases remain elusive, Cx's are generally considered to mediate cell-to-cell communication by forming gap junction channels. We show here that unlike rat Cx26, human and sheep Cx26 form voltage-gated hemichannels when expressed in oocytes and Neuro2A cells. A single evolutionary amino acidic change at position 159 of the rodent protein, the replacement of aspartic acid with asparagine in the human and sheep proteins, accounts for this species specificity. At the resting potential and in normal millimolar extracellular calcium, open human Cx26 hemichannels can be detected both electrophysiologically and by dye uptake, although they did not affect cell viability. These hemichannels opened at approximately -50 mV and their activation increased by depolarization until they inactivate at positive membrane potentials. Single channel analysis revealed that activation and inactivation involved two distinct voltage gating mechanisms and that the fully open hemichannel displays a conductance twice that of the intercellular channel. The existence of a hemichannel that opens under physiological control of the membrane potential may have important implications for the normal and pathological activity of Cx26 in humans, particularly with respect to hearing and the epidermis. PMID- 17077311 TI - Design of phosphorylated dendritic architectures to promote human monocyte activation. AB - As first defensive line, monocytes are a pivotal cell population of innate immunity. Monocyte activation can be relevant to a range of immune conditions and responses. Here we present new insights into the activation of monocytes by a series of phosphonic acid-terminated, phosphorus-containing dendrimers. Various dendritic or subdendritic structures were synthesized and tested, revealing the basic structural requirements for monocyte activation. We showed that multivalent character and phosphonic acid capping of dendrimers are crucial for monocyte targeting and activation. Confocal videomicroscopy showed that a fluorescein tagged dendrimer binds to isolated monocytes and gets internalized within a few seconds. We also found that dendrimers follow the phagolysosomial route during internalization by monocytes. Finally, we performed fluorescence resonance energy transfer (FRET) experiments between a specifically designed fluorescent dendrimer and phycoerythrin-coupled antibodies. We showed that the typical innate Toll-like receptor (TLR)-2 is clearly involved, but not alone, in the sensing of dendrimers by monocytes. In conclusion, phosphorus-containing dendrimers appear as precisely tunable nanobiotools able to target and activate human innate immunity and thus prove to be good candidates to develop new drugs for immunotherapies. PMID- 17077312 TI - Evidence for local relaxin ligand-receptor expression and function in arteries. AB - Relaxin is a 6 kDa protein hormone produced by the corpus luteum and secreted into the blood during pregnancy in rodents and humans. Growing evidence indicates that circulating relaxin causes vasodilatation and increases in arterial compliance, which may be among its most important actions during pregnancy. Here we investigated whether there is local expression and function of relaxin and relaxin receptor in arteries of nonpregnant females and males. Relaxin-1 and its major receptor, Lgr7, mRNA are expressed in thoracic aortas, small renal and mesenteric arteries from mice and rats of both sexes, as well as in small renal arteries from female tammar wallabies (an Australian marsupial). Using available antibodies for rat and mouse Lgr7 receptor and rat relaxin, we also identified protein expression in arteries. Small renal arteries isolated from relaxin-1 gene deficient mice demonstrate enhanced myogenic reactivity and decreased passive compliance relative to wild-type (WT) and heterozygous mice. Taken together, these findings reveal an arterial-derived, relaxin ligand-receptor system that acts locally to regulate arterial function. PMID- 17077313 TI - Abnormalities in the pattern of AQP4 immunoreactivity. PMID- 17077315 TI - Up-regulated PAR-2-mediated salivary secretion in mice deficient in muscarinic acetylcholine receptor subtypes. AB - Protease-activated receptor-2 (PAR-2) is expressed in the salivary glands and is expected to be a new target for the treatment of exocrine dysfunctions, such as dry mouth; however, the salivary secretory mechanism mediated by PAR-2 remains to be elucidated. Therefore, mechanism of the PAR-2-mediated salivary secretion was investigated in this study. We found that a PAR-2 agonist peptide, SLIGRL-OH, induced salivary flow in vivo and dose-dependent increase in [Ca(2+)](i) submandibular gland (SMG) acinar cells in wild-type (WT) mice and mice lacking M(3) or both M(1) and M(3) muscarinic acetylcholine receptors (mAChRs), whereas secretions in PAR-2 knockout (PAR-2KO) mice were completely abolished. The saliva composition secreted by SLIGRL-OH was similar to that secreted by mAChR stimulation. Ca(2+) imaging in WT acinar cells and beta-galactosidase staining in PAR-2KO mice, in which the beta-galactosidase gene (LacZ) was incorporated into the disrupted gene, revealed a nonubiquitous, sporadic distribution of PAR-2 in the SMG. Furthermore, compared with the secretion in WT mice, PAR-2-mediated salivary secretion and Ca(2+) response were enhanced in mice lacking M(3) or both M(1) and M(3) mAChRs, in which mAChR-stimulated secretion and Ca(2+) response in acinar cells were severely impaired. Although the mechanism underlying the enhanced PAR-2-mediated salivary secretion in M(3)-deficient mice is not clear, the result suggests the presence of some compensatory mechanism involving PAR-2 in the salivary glands deficient in cholinergic activation. These results indicate that PAR-2 present in the salivary glands mediates Ca(2+)-dependent fluid secretion, demonstrating potential usefulness of PAR-2 as a target for dry mouth treatment. PMID- 17077316 TI - Dexamethasone inhibits epidermal growth factor-stimulated gastric epithelial cell proliferation. AB - Epidermal growth factor (EGF) is essential to heal gastric ulcers, whereas glucocorticoid delays rat gastric ulcer healing. We found that dexamethasone inhibited EGF-stimulated rat gastric epithelial cell (RGM-1) proliferation by cell count and DNA synthesis analysis of flow cytometry and attempted to elucidate the possible mechanistic pathway via Western blot analysis. EGF (10 ng/ml) treatment for 24 h significantly increased RGM-1 cell proliferation, and dexamethasone (10(-8) and 10(-6) M) markedly suppressed EGF-stimulated cell proliferation. Western blotting results demonstrated that the phosphorylated extracellular signal-regulated kinase (pERK) (pERK1/pERK2) significantly increased at 10 min after EGF treatment. This was followed by increase of cyclooxygenase (COX)-2 expression at 3 h after EGF treatment. The continued increase of COX-2 (up to 18 h) resulted in increased intracellular prostaglandin E(2) and cyclin D1 expression significantly after 8 and 12 h of EGF treatment. Dexamethasone substantially reduced EGF-stimulated COX-2 expression at 3 and 6 h and cyclin D1 expression at 8 and 12 h. Pretreatment of RGM-1 cells with dexamethasone or 2'-amino-3'-methoxyflavone (PD98059)-mitogen-activated protein kinase kinase inhibitor (5 x 10(-5) M) significantly reduced EGF-stimulated pERK1/pERK2 expression. Simultaneous treatment of RGM-1 cells with PD98059 and EGF also markedly decreased EGF-stimulated COX-2 expression at 6 h. These findings indicate that dexamethasone significantly suppresses EGF-stimulated gastric epithelial cell proliferation, and one of the pathways involved is via inhibiting activation of ERK1/ERK2, followed by inhibition of COX-2, cyclin D1 expression, and finally DNA synthesis. PMID- 17077317 TI - 5-Hydroxytryptamine (serotonin)2A receptors in rat anterior cingulate cortex mediate the discriminative stimulus properties of d-lysergic acid diethylamide. AB - d-Lysergic acid diethylamide (LSD), an indoleamine hallucinogen, produces profound alterations in mood, thought, and perception in humans. The brain site(s) that mediates the effects of LSD is currently unknown. In this study, we combine the drug discrimination paradigm with intracerebral microinjections to investigate the anatomical localization of the discriminative stimulus of LSD in rats. Based on our previous findings, we targeted the anterior cingulate cortex (ACC) to test its involvement in mediating the discriminative stimulus properties of LSD. Rats were trained to discriminate systemically administered LSD (0.085 mg/kg s.c.) from saline. Following acquisition of the discrimination, bilateral cannulae were implanted into the ACC (AP, +1.2 mm; ML, +/-1.0 mm; DV, -2.0 mm relative to bregma). Rats were tested for their ability to discriminate varying doses of locally infused LSD (0.1875, 0.375, and 0.75 microg/side) or artificial cerebrospinal fluid (n = 3-7). LSD locally infused into ACC dose-dependently substituted for systemically administered LSD, with 0.75 microg/side LSD substituting completely (89% correct). Systemic administration of the selective 5 hydroxytryptamine (serotonin) (5-HT)(2A) receptor antagonist R-(+)-alpha-(2,3 dimethoxyphenyl)-1-[2-(4-fluorophenylethyl)]-4-piperidine-methanol (M100907; 0.4 mg/kg) blocked the discriminative cue of LSD (0.375 microg/side) infused into ACC (from 68 to 16% drug lever responding). Furthermore, M100907 (0.5 microg/microl/side) locally infused into ACC completely blocked the stimulus effects of systemic LSD (0.04 mg/kg; from 80 to 12% on the LSD lever). Taken together, these data indicate that 5-HT(2A) receptors in the ACC are a primary target mediating the discriminative stimulus properties of LSD. PMID- 17077318 TI - ZM241385, DPCPX, MRS1706 are inverse agonists with different relative intrinsic efficacies on constitutively active mutants of the human adenosine A2B receptor. AB - The human adenosine A(2B) receptor belongs to class A G protein-coupled receptors (GPCRs). In our previous work, constitutively active mutant (CAM) human adenosine A(2B) receptors were identified from a random mutation bank. In the current study, three known A(2B) receptor antagonists, 4-{2-[7-amino-2-(2 furyl)[1,2,4]triazolo-[2,3-a][1,3,5]triazin-5-yl-amino]ethyl}phenol (ZM241385), 8 cyclopentyl-1,3-dipropylxanthine (DPCPX), and N-(4-acetylphenyl)-2-[4-(2,3,6,7 tetrahydro-2,6-dioxo-1,3-dipropyl-1H-purin-8-yl)phenoxy]acetamide (MRS1706) were tested on wild-type and nine CAM A(2B) receptors with different levels of constitutive activity in a yeast growth assay. All three compounds turned out to be inverse agonists for the adenosine A(2B) receptor because they were able to fully reverse the basal activity of four low-level constitutively active A(2B) receptor mutants and to partially reverse the basal activity of three medium level constitutively active A(2B) receptor mutants. We also discovered two highly constitutively active mutants whose basal activity could not be reversed by any of the three compounds. A two-state receptor model was used to explain the experimental observations; fitting these yielded the following relative intrinsic efficacies for the three inverse agonists ZM241385, DPCPX, and MRS1706: 0.14 +/- 0.03, 0.35 +/- 0.03, and 0.31 +/- 0.02, respectively. Moreover, varying L, the ratio of active versus inactive receptors in this model, from 0.11 for mutant F84L to 999 for two highly constitutively active mutants yielded simulated dose response curves that mimicked the experimental curves. This study is the first description of inverse agonists for the human adenosine A(2B) receptor. Moreover, the use of receptor mutants with varying levels of constitutive activity enabled us to determine the relative intrinsic efficacy of these inverse agonists. PMID- 17077319 TI - Identification of an infectious progenitor for the multiple-copy HERV-K human endogenous retroelements. AB - Human Endogenous Retroviruses are expected to be the remnants of ancestral infections of primates by active retroviruses that have thereafter been transmitted in a Mendelian fashion. Here, we derived in silico the sequence of the putative ancestral "progenitor" element of one of the most recently amplified family - the HERV-K family - and constructed it. This element, Phoenix, produces viral particles that disclose all of the structural and functional properties of a bona-fide retrovirus, can infect mammalian, including human, cells, and integrate with the exact signature of the presently found endogenous HERV-K progeny. We also show that this element amplifies via an extracellular pathway involving reinfection, at variance with the non-LTR-retrotransposons (LINEs, SINEs) or LTR-retrotransposons, thus recapitulating ex vivo the molecular events responsible for its dissemination in the host genomes. We also show that in vitro recombinations among present-day human HERV-K (also known as ERVK) loci can similarly generate functional HERV-K elements, indicating that human cells still have the potential to produce infectious retroviruses. PMID- 17077320 TI - Transcriptional interference among the murine beta-like globin genes. AB - Mammalian beta-globin loci contain multiple genes that are activated at different developmental stages. Studies have suggested that the transcription of one gene in a locus can influence the expression of the other locus genes. The prevalent model to explain this transcriptional interference is that all potentially active genes compete for locus control region (LCR) activity. To investigate the influence of transcription by the murine embryonic genes on transcription of the other beta-like genes, we generated mice with deletions of the promoter regions of Ey and betah1 and measured transcription of the remaining genes. Deletion of the Ey and betah1 promoters increased transcription of betamajor and betaminor 2 fold to 3-fold during primitive erythropoiesis. Deletion of Ey did not affect betah1 nor did deletion of betah1 affect Ey, but Ey deletion uniquely activated transcription from betah0, a beta-like globin gene immediately downstream of Ey. Protein analysis showed that betah0 encodes a translatable beta-like globin protein that can pair with alpha globin. The lack of transcriptional interference between Ey and betah1 and the gene-specific repression of betah0 did not support LCR competition among the embryonic genes and suggested that direct transcriptional interference from Ey suppressed betah0. PMID- 17077321 TI - Evidence for the multimeric structure of ferroportin. AB - Ferroportin (Fpn) (IREG1, SLC40A1, MTP1) is an iron transporter, and mutations in Fpn result in a genetically dominant form of iron overload disease. Previously, we demonstrated that Fpn is a multimer and that mutations in Fpn are dominant negative. Other studies have suggested that Fpn is not a multimer and that overexpression or epitope tags might affect the localization, topology, or multimerization of Fpn. We generated wild-type Fpn with 3 different epitopes, GFP, FLAG, and c-myc, and expressed these constructs in cultured cells. Co expression of any 2 different epitope-tagged proteins in the same cell resulted in their quantitative coimmunoprecipitation. Treatment of Fpn-GFP/Fpn-FLAG expressing cells with crosslinking reagents resulted in the crosslinking of Fpn GFP and Fpn-FLAG. Western analysis of rat glioma C6 cells or mouse bone marrow macrophages exposed to crosslinking reagents showed that endogenous Fpn is a dimer. These results support the hypothesis that the dominant inheritance of Fpn iron overload disease is due to the dominant-negative effects of mutant Fpn proteins. PMID- 17077322 TI - Expression of the granzyme B inhibitor PI9 predicts outcome in nasal NK/T-cell lymphoma: results of a Western series of 48 patients treated with first-line polychemotherapy within the Groupe d'Etude des Lymphomes de l'Adulte (GELA) trials. AB - Nasal NK/T-cell lymphoma is a rare disease entity with a poor outcome. Expression of antiapoptotic proteins has not been extensively investigated in this entity. Forty-eight patients with nasal T/NK-cell lymphoma who received first-line polychemotherapy (n = 44) or chemoradiotherapy (n = 4) were analyzed for expression of active caspase-3 (aC3), granzyme B protease inhibitor 9 (PI9), and Bcl-2 proteins. Lymphomas were CD3+/CD5-/granzyme B+ and EBV-associated. Median age was 46 years. Stage I/II disease was present in 75% of the cases and an International Prognostic Index (IPI) score less than 1 in 65%. With a median follow-up of 6.3 years, 5-year event-free survival (EFS) and overall survival (OS) rates were 39% and 49%, respectively. Apoptotic index was scored as high in 32% of cases and PI9 expression as positive in 68%, whereas 35% disclosed a high number of aC3+ tumor cells. Univariate analysis showed that absence of PI9 and low apoptotic index were associated with poor outcome, but not aC3 expression nor IPI score. By multivariate analysis, both parameters affected independently EFS (P = .02 and .08, respectively) and OS (P = .009 and .04). In view of its constitutive expression by normal NK cells, it is suggested that loss of PI9 expression in tumor cells may reflect some mechanism associated with progression. PMID- 17077323 TI - Bone morphogenic protein antagonist Drm/gremlin is a novel proangiogenic factor. AB - Angiogenesis plays a key role in various physiologic and pathologic conditions, including tumor growth. Drm/gremlin, a member the Dan family of bone morphogenic protein (BMP) antagonists, is commonly thought to affect different processes during growth, differentiation, and development by heterodimerizing various BMPs. Here, we identify Drm/gremlin as a novel proangiogenic factor expressed by endothelium. Indeed, Drm/gremlin was purified to homogeneity from the conditioned medium of transformed endothelial cells using an endothelial-cell sprouting assay to follow protein isolation. Accordingly, recombinant Drm/gremlin stimulates endothelial-cell migration and invasion in fibrin and collagen gels, binds with high affinity to various endothelial cell types, and triggers tyrosine phosphorylation of intracellular signaling proteins. Also, Drm/gremlin induces neovascularization in the chick embryo chorioallantoic membrane. BMP4 does not affect Drm/gremlin interaction with endothelium, and both molecules exert a proangiogenic activity in vitro and in vivo when administered alone or in combination. Finally, Drm/gremlin is produced by the stroma of human tumor xenografts in nude mice, and it is highly expressed in endothelial cells of human lung tumor vasculature when compared with non-neoplastic lung. Our observations point to a novel, previously unrecognized capacity of Drm/gremlin to interact directly with target endothelial cells and to modulate angiogenesis. PMID- 17077324 TI - Endolyn (CD164) modulates the CXCL12-mediated migration of umbilical cord blood CD133+ cells. AB - Hematopoietic stem cell/hematopoietic progenitor cell (HSC/HPC) homing to specific microenvironmental niches involves interactions between multiple receptor ligand pairs. Although CXCL12/CXCR4 plays a central role in these events, CXCR4 regulators that provide the specificity for such cells to lodge and be retained in particular niches are poorly defined. Here, we provide evidence that the sialomucin endolyn (CD164), an adhesion receptor that regulates the adhesion of CD34+ cells to bone marrow stroma and the recruitment of CD34+CD38(lo/-) cells into cycle, associates with CXCR4. The class II 103B2 monoclonal antibody, which binds the CD164 N-linked glycan-dependent epitope or CD164 knockdown by RNA interference, significantly inhibits the migration of CD133+ HPCs toward CXCL12 in vitro. On presentation of CXCL12 on fibronectin, CD164 associates with CXCR4, an interaction that temporally follows the association of CXCR4 with the integrins VLA-4 and VLA-5. This coincides with PKC zeta and Akt signaling through the CXCR4 receptor, which was disrupted on the loss of CD164 though MAPK signaling was unaffected. We therefore demonstrate a novel association among 3 distinct families of cell-surface receptors that regulate cell migratory responses and identify a new role for CD164. We propose that this lends specificity to the homing and lodgment of these cells within the bone marrow niche. PMID- 17077325 TI - ATM deficiency disrupts Tcra locus integrity and the maturation of CD4+CD8+ thymocytes. AB - Mutations in ATM (ataxia-telangiectasia mutated) cause ataxia-telangiectasia (AT), a disease characterized by neurodegeneration, sterility, immunodeficiency, and T-cell leukemia. Defective ATM-mediated DNA damage responses underlie many aspects of the AT syndrome, but the basis for the immune deficiency has not been defined. ATM associates with DNA double-strand breaks (DSBs), and some evidence suggests that ATM may regulate V(D)J recombination. However, it remains unclear how ATM loss compromises lymphocyte development in vivo. Here, we show that T cell receptor beta (TCRbeta)-dependent proliferation and production of TCRbeta(low) CD4+CD8+ (DP) thymocytes occurred normally in Atm-/- mice. In striking contrast, the postmitotic maturation of TCRbeta(low) DP precursors into TCRbeta(int) DP cells and TCRbeta(hi) mature thymocytes was profoundly impaired. Furthermore, Atm-/- thymocytes expressed abnormally low amounts of TCRalpha mRNA and protein. These defects were not attributable to the induction of a BCL-2 sensitive apoptotic pathway. Rather, they were associated with frequent biallelic loss of distal Va gene segments in DP thymocytes, revealing that ATM maintains Tcra locus integrity as it undergoes V(D)J recombination. Collectively, our data demonstrate that ATM loss increases the frequency of aberrant Tcra deletion events, which compromise DP thymocyte maturation and likely promote the generation of oncogenic TCR translocations. PMID- 17077326 TI - Gene-expression profiling of systemic anaplastic large-cell lymphoma reveals differences based on ALK status and two distinct morphologic ALK+ subtypes. AB - With the use of microarray gene-expression profiling, we analyzed a homogeneous series of 32 patients with systemic anaplastic large-cell lymphoma (ALCL) and 5 ALCL cell lines. Unsupervised analysis classified ALCL in 2 clusters, corresponding essentially to morphologic subgroups (ie, common type vs small cell and "mixed" variants) and clinical variables. Patients with a morphologic variant of ALCL had advanced-stage disease. This group included a significant number of patients who experienced early relapse. Supervised analysis showed that ALK+ALCL and ALK- ALCL have different gene-expression profiles, further confirming that they are different entities. Among the most significantly differentially expressed genes between ALK+ and ALK- samples, we found BCL6, PTPN12, CEBPB, and SERPINA1 genes to be overexpressed in ALK+ ALCL. This result was confirmed at the protein level for BCL-6, C/EBPbeta and serpinA1 through tissue microarrays. The molecular signature of ALK- ALCL included overexpression of CCR7, CNTFR, IL22, and IL21 genes but did not provide any obvious clues to the molecular mechanism underlying this tumor subtype. Once confirmed on a larger number of patients, the results of the present study could be used for clinical and therapeutic management of patients at the time of diagnosis. PMID- 17077327 TI - FGF2 posttranscriptionally down-regulates expression of SDF1 in bone marrow stromal cells through FGFR1 IIIc. AB - The chemokine stromal cell-derived factor-1 (SDF-1) is constitutively expressed by bone marrow stromal cells and plays key roles in hematopoiesis. Fibroblast growth factor 2 (FGF2), a member of the FGF family that plays important roles in developmental morphogenic processes, is abnormally elevated in the bone marrow from patients with clonal myeloid disorders and other disorders where normal hematopoiesis is impaired. Here, we report that FGF2 reduces SDF-1 secretion and protein content in bone marrow stromal cells. By inhibiting SDF-1 production, FGF2 compromises stromal cell support of hematopoietic progenitor cells. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis revealed that bone marrow stromal cells express 5 FGF receptors (FGFRs) among the 7 known FGFR subtypes. Blocking experiments identified FGFR1 IIIc as the receptor mediating FGF2 inhibition of SDF-1 expression in bone marrow stromal cells. Analysis of the mechanisms underlying FGF2 inhibition of SDF-1 production in bone marrow stromal cells revealed that FGF2 reduces the SDF-1 mRNA content by posttranscriptionally accelerating SDF-1 mRNA decay. Thus, we identify FGF2 as an inhibitor of SDF-1 production in bone marrow stromal cells and a regulator of stromal cell supportive functions for hematopoietic progenitor cells. PMID- 17077328 TI - MEK blockade converts AML differentiating response to retinoids into extensive apoptosis. AB - The aberrant function of transcription factors and/or kinase-based signaling pathways that regulate the ability of hematopoietic cells to proliferate, differentiate, and escape apoptosis accounts for the leukemic transformation of myeloid progenitors. Here, we demonstrate that simultaneous retinoid receptor ligation and blockade of the MEK/ERK signaling module, using the small-molecule inhibitor CI-1040, result in a strikingly synergistic induction of apoptosis in both acute myeloid leukemia (AML) and acute promyelocytic leukemia (APL) cells with constitutive ERK activation. This proapoptotic synergism requires functional RAR and RXR retinoid receptors, as demonstrated using RAR- and RXR-selective ligands and RAR-defective cells. In the presence of MEK inhibitors, however, retinoid-induced chromatin remodeling, target-gene transcription, and granulocytic differentiation are strikingly inhibited and apoptosis induction becomes independent of death-inducing ligand/receptor pairs; this suggests that apoptosis induction by combined retinoids and MEK inhibitors is entirely distinct from the classical "postmaturation" apoptosis induced by retinoids alone. Finally, we identify disruption of Bcl-2-dependent mitochondrial homeostasis as a possible point of convergence for the proapoptotic synergism observed with retinoids and MEK inhibitors. Taken together, these results indicate that combined retinoid treatment and MEK blockade exert powerful antileukemic effects and could be developed into a novel therapeutic strategy for both AML and APL. PMID- 17077329 TI - Immune regulation by pretenders: cell-to-cell transfers of HLA-G make effector T cells act as regulatory cells. AB - Trogocytosis is the uptake of membrane fragments from one cell by another and has been described for immune cells in mice and humans. Functional consequences of trogocytosis are emerging, but a dramatic immune function has still to be associated with it. Here we show that some resting, and most activated, CD4+ and CD8+ T cells acquire immunosuppressive HLA-G1 from antigen-presenting cells (APCs) in a few minutes. Acquisition of HLA-G through membrane transfers does not change the real nature of the T cells but immediately reverses their function from effectors to regulatory cells. These regulatory cells can inhibit allo proliferative responses through HLA-G1 that they acquired. These data demonstrate that trogocytosis of HLA-G1 leads to instant generation of a new type of regulatory cells, which act through cell-surface molecules they temporarily display but do not express themselves. Such regulatory cells whose existence is most likely limited in space and time might constitute an "emergency" immune suppression mechanism used by HLA-G-expressing tissues to protect themselves against immune aggression. In addition, T cells acquire from HLA-G-expressing APCs their HLA-G-dependent capability to induce the slower differentiation of regulatory cells that act independently of HLA-G. These data re-emphasize the significance of HLA-G expression in normal and pathologic situations. PMID- 17077330 TI - IL-21 receptor signaling is integral to the development of Th2 effector responses in vivo. AB - Interleukin 21 (IL-21) is a member of the common gamma-chain family of cytokines, which influence a broad spectrum of immunologic responses. A number of studies have examined the function of IL-21, but its specific role in Th1/Th2-cell differentiation and related effector responses remains to be clarified. Thus, we generated IL-21R-deficient mice and have investigated the role of IL-21R signaling using a series of in vivo experimentally induced disease models. We first addressed the role of IL-21R signaling in Th2 immune responses by examining allergic airway inflammation, and Nippostrongylus brasiliensis and Heligmosomoides polygyrus antihelminth responses. In each of these systems, IL 21R signaling played a clear role in the development of Th2 responses. Comparatively, IL-21R signaling was not required for the containment of Leishmania major infection or the development of experimental autoimmune myocarditis, indicative of competent Th1 and Th17 responses, respectively. Adoptive transfer of T cells and analysis of IL-21R+/+/IL-21R-/- chimera mice revealed that IL-21R-signaling was central to Th2-cell survival or migration to peripheral tissues. Overall, our data show IL-21 plays a crucial role in supporting polarized Th2 responses in vivo, while appearing superfluous for Th1 and Th17 responses. PMID- 17077331 TI - 4q loss is potentially an important genetic event in MM tumorigenesis: identification of a tumor suppressor gene regulated by promoter methylation at 4q13.3, platelet factor 4. AB - In this study, we have elucidated the chromosomal imbalances in the multistep pathogenesis and delineated several critical tumor suppressor gene (TSG) loci in multiple myeloma (MM). By using comparative genomic hybridization, allelotyping, and multicolor interphase fluorescence in situ hybridization, 5 MM cell lines and bone marrow CD138+ plasma cells from 88 Chinese patients with monoclonal gammopathy of undetermined significance (MGUS) and early and advanced stages of MM were investigated. In all MGUS and MM samples, chromosome copy number abnormalities were detected. A higher number of chromosomal imbalances and specific genetic alterations are involved in MGUS to MM transition (-6q, +3p, and +1p) and MM progression (+2p and +9q). In addition to -13q, we first found high frequencies (42% to 46%) of -4q involving high percentages (70% to 74%) of clonal plasma cells in both MGUS and MM, suggesting that inactivation of TSG in this region is also a potentially critical genetic event in MM tumorigenesis. By high resolution allelotyping, we defined a common deletion region on 4q13.3 and found that a candidate TSG, platelet factor 4, was frequently silenced by promoter hypermethylation in MM (15 of 28) and MM cell lines (5 of 5). These data have opened up a new approach in the molecular targeting therapy and provide novel insights into MM tumorigenesis. PMID- 17077332 TI - Emodin and DHA potently increase arsenic trioxide interferon-alpha-induced cell death of HTLV-I-transformed cells by generation of reactive oxygen species and inhibition of Akt and AP-1. AB - Adult T-cell leukemia (ATL) is an aggressive lymphoproliferative disease of poor clinical prognosis associated with infection by the human T-cell leukemia virus type I (HTLV-I). The use of arsenic trioxide (As2O3) has been shown to effectively treat acute promyelocytic leukemia (APL) with greater than 80% of patients achieving complete remission. The combination of arsenic and interferon has also shown promising results in the treatment of ATL. The requirement for slow dosage increases of arsenic and the time required to achieve a pharmacologic active dose in patients is a major obstacle because median survival of patients with ATL is about 6 months. In this study we report a potent synergistic effect of the combination of arsenic trioxide and interferon alpha (As/IFN-alpha) with emodin and DHA on cell-cycle arrest and cell death of HTLV-I-infected cells. Importantly, we found that clinically achievable doses of DHA and emodin allowed for reduced arsenic concentrations by 100-fold while still remaining highly toxic to tumor cells. Our data provide a rationale for combined use of As/IFN-alpha with emodin and DHA in patients with ATL refractory to conventional therapy. PMID- 17077333 TI - Therapy with high-dose dexamethasone (HD-DXM) in previously untreated patients affected by idiopathic thrombocytopenic purpura: a GIMEMA experience. AB - In idiopathic thrombocytopenic purpura (ITP), corticosteroids have been widely recognized as the most appropriate first-line treatment, even if the best therapeutic approach is still a matter of debate. Recently, a single high-dose dexamethasone (HD-DXM) course was administered as first-line therapy in adult patients with ITP. In this paper we show the results of 2 prospective pilot studies (monocentric and multicentric, respectively) concerning the use of repeated pulses of HD-DXM in untreated ITP patients. In the monocenter study, 37 patients with severe ITP, age at least 20 years and no more than 65 years, were enrolled. HD-DXM was given in 4-day pulses every 28 days, for 6 cycles. Response rate was 89.2%; relapse-free survival (RFS) was 90% at 15 months; long-term responses, lasting for a median time of 26 months (range 6-77 months) were 25 of 37 (67.6%). In the multicenter study, 95 patients with severe ITP, age at least 2 years and no more than 70 years, were enrolled. HD-DXM was given in 4-day pulses every 14 days, for 4 cycles; 90 patients completed 4 cycles. Response rate (85.6%) was similar in patients classified by age (<18 years, 36 of 42=85.7%; >or=18 years, 41 of 48=85.4%, P=not significant), with a statistically significant difference between the second and third cycle (75.8% vs 89%, P=.018). RFS at 15 months 81%; long-term responses, lasting for a median time of 8 months (range 4-24 months) were 67 of 90 (74.4%). In both studies, therapy was well tolerated. A schedule of 3 cycles of HD-DXM pulses will be compared with standard prednisone therapy (eg, 1 mg/kg per day) in the next randomized Gruppo Italiano Malattie EMatologiche dell'Adulto (GIMEMA) trial. PMID- 17077334 TI - Patient-centered care. PMID- 17077335 TI - Bisphosphonate use in children with bone disease. PMID- 17077337 TI - Applications of porous tantalum in total hip arthroplasty. AB - Porous tantalum is an alternative metal for total joint arthroplasty components that offers several unique properties. Its high volumetric porosity (70% to 80%), low modulus of elasticity (3 MPa), and high frictional characteristics make it conducive to biologic fixation. Tantalum has excellent biocompatibility and is safe to use in vivo. The low modulus of elasticity allows for more physiologic load transfer and relative preservation of bone stock. Because of its bioactive nature and ingrowth properties, tantalum is used in primary as well as revision total hip arthroplasty components, with good to excellent early clinical results. In revision arthroplasty, standard and custom augments may serve as a structural bone graft substitute. Formation of a bone-like apatite coating in vivo affords strong fibrous ingrowth properties and allows for substantial soft-tissue attachment, indicating potential for use in cases requiring reattachment of muscles and tendons to a prosthesis. Development of modular components and femoral stems also is being evaluated. The initial clinical data and basic science studies support further investigation of porous tantalum as an alternative to traditional implant materials. PMID- 17077338 TI - Sacral fractures. AB - Sacral fractures most commonly occur after pelvic ring injuries but occasionally in isolation. Although the true incidence of sacral fractures is unknown, an estimated 30% are identified late. Sequelae of inappropriately treated or untreated sacral fractures include persistent pain, decreased mobility, and neurologic compromise. Because these fractures often result from high-energy trauma, concomitant injuries should be suspected. A thorough physical examination, including a detailed neurologic assessment and radiographic evaluation, is necessary to determine treatment. Computed tomography of the pelvis/sacrum can provide significant information about fracture pattern. Surgical intervention, often as a combination of neural decompression and stabilization, is indicated in patients with neurologic deficits, significant soft-tissue compromise, and lumbosacral instability. Patient satisfaction with surgical intervention has not been definitively documented, although neurologic improvement with timely intervention has been noted. PMID- 17077339 TI - Slipped capital femoral epiphysis: current concepts. AB - Slipped capital femoral epiphysis is a common hip disorder in adolescents, with an incidence of 0.2 (Japan) to 10 (United States) per 100,000. The etiology is unknown, but biomechanical and biochemical factors play an important role. Symptoms at presentation include pain in the groin, thigh, or knee. Ambulatory patients also may present with a limp. Nonambulatory patients present with excruciating pain. The slipped capital femoral epiphysis is classified as stable when the patient can walk and unstable when the patient cannot walk, even with the aid of crutches. Because the epiphysis slips posteriorly, it is best seen on lateral radiographs. The treatment of choice for stable slipped capital femoral epiphysis is single-screw fixation in situ. This method has a high probability of long-term success, with minimal risk of complications. In the patient with unstable slipped capital femoral epiphysis, urgent hip joint aspiration followed by closed reduction and single- or double-screw fixation provides the best environment for a satisfactory result, while minimizing the risk of complications. PMID- 17077340 TI - Malignant tumors of the hand and wrist. AB - Malignant tumors in the hand and wrist compose a wide variety of lesions involving skin, soft tissues, and bone. Although these lesions are found elsewhere in the body, many have unique characteristics at this anatomic location. Skin tumors predominate; the most common are squamous cell carcinomas, followed in frequency by basal cell carcinomas and malignant melanomas. Other soft-tissue malignancies are less common but may present more difficult diagnostic problems. They often appear as painless masses that sometimes have been present for months or even years and deceptively appear to be benign. A missed or delayed diagnosis of these tumors can have devastating consequences. Bone malignancies involve both primary lesions, of which chondrosarcomas are the most common, and metastatic lesions. Regardless of cell type, treatment of malignant tumors in the hand and wrist requires special considerations because of the important function of these structures. Orthopaedic surgeons should be familiar with the spectrum of these tumors, the work-up necessary to arrive at a precise diagnosis, and the treatment that will achieve the most favorable outcome. PMID- 17077341 TI - Elbow valgus instability in the throwing athlete. PMID- 17077342 TI - Glucocorticoids produce whole body insulin resistance with changes in cardiac metabolism. AB - Insulin resistance is viewed as an insufficiency in insulin action, with glucocorticoids being recognized to play a key role in its pathogenesis. With insulin resistance, metabolism in multiple organ systems such as skeletal muscle, liver, and adipose tissue is altered. These metabolic alterations are widely believed to be important factors in the morbidity and mortality of cardiovascular disease. More importantly, clinical and experimental studies have established that metabolic abnormalities in the heart per se also play a crucial role in the development of heart failure. Following glucocorticoids, glucose utilization is compromised in the heart. This attenuated glucose metabolism is associated with altered fatty acid supply, composition, and utilization. In the heart, elevated fatty acid use has been implicated in a number of metabolic, morphological, and mechanical changes and, more recently, in "lipotoxicity". In the present article, we review the action of glucocorticoids, their role in insulin resistance, and their influence in modulating peripheral and cardiac metabolism and heart disease. PMID- 17077343 TI - Ovariectomy worsens secondary hyperparathyroidism in mature rats during low-Ca diet. AB - Estrogen deficiency impairs intestinal Ca absorption and induces bone loss, but its effects on the vitamin D-endocrine system are unclear. In the present study, calciotropic hormones levels, renal vitamin D metabolism, 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]-dependent intestinal calcium absorption, and bone properties in 3-mo-old sham-operated (sham) or ovariectomized (OVX) rats fed either a normal-Ca (NCD; 0.6% Ca, 0.65% P) or a low-Ca (LCD; 0.1% Ca, 0.65% P) diet for 2 wk were determined. LCD increased serum 1,25(OH)2D3 levels in both sham and OVX rats. Serum parathyroid hormone [PTH(1-84)] levels were highest in OVX rats fed LCD. Renal 25-hydroxyvitamin D1alpha-hydroxylase (1-OHase) protein expression was induced in both sham and OVX rats during LCD, while renal 1-OHase mRNA expression was highest in OVX rats fed LCD. Renal vitamin D receptor (VDR) and mRNA expressions in rats were induced by ovariectomy in rats fed NCD but suppressed by ovariectomy in rats fed LCD. The induction of intestinal calcium transporter-1 and calbindin-D9k mRNA expressions by LCD were not altered by ovariectomy. As expected, bone Ca content, cancellous bone mineral density, and bone strength index in proximal metaphysis of rat tibia were reduced by both ovariectomy and LCD (P<0.05) as analyzed by two-way ANOVA. Taken together, the data demonstrate that ovariectomy alters the responses of circulating PTH levels, renal 1-OHase mRNA expression, and renal VDR expression to LCD. These results suggest that estrogen is necessary for the full adaptive response to LCD mediated by both PTH and 1,25(OH)2D3. PMID- 17077344 TI - AS160 phosphorylation is associated with activation of alpha2beta2gamma1- but not alpha2beta2gamma3-AMPK trimeric complex in skeletal muscle during exercise in humans. AB - We investigated time- and intensity-dependent effects of exercise on phosphorylation of Akt substrate of 160 kDa (AS160) in human skeletal muscle. Subjects performed cycle exercise for 90 min (67% VO2 peak, n=8), 20 min (80% VO2 peak, n=11), 2 min (110% of peak work rate, n=9), or 30 s (maximal sprint, n=10). Muscle biopsies were obtained before, during, and after exercise. In trial 1, AS160 phosphorylation increased at 60 min (60%, P=0.06) and further at 90 min of exercise (120%, P<0.05). alpha2beta2gamma3-AMP-activated protein kinase (AMPK) activity increased significantly to a steady-state level after 30 min, whereas alpha2beta2gamma1-AMPK activity increased after 60 min of exercise with a further significant increase after 90 min. alpha2beta2gamma1-AMPK activity and AS160 phosphorylation correlated positively (r2=0.55). In exercise trials 2, 3, and 4, alpha2beta2gamma3-AMPK activity but neither AS160 phosphorylation nor alpha2beta2gamma1-AMPK activity increased. Akt Ser473 phosphorylation was unchanged in all trials, whereas Akt Thr308 phosphorylation increased significantly in trial 3 and 4 only. These results show that AS160 is phosphorylated in a time-dependent manner during moderate-intensity exercise and suggest that alpha2beta2gamma1- but not alpha2beta2gamma3-AMPK may act in a pathway responsible for exercise-induced AS160 phosphorylation. Furthermore, we show that AMPK complexes in skeletal muscle are activated differently depending on exercise intensity and duration. PMID- 17077345 TI - Pentoxifylline inhibits Ca2+-dependent and ATP proteasome-dependent proteolysis in skeletal muscle from acutely diabetic rats. AB - Previous studies from this laboratory have shown that catecholamines exert an inhibitory effect on muscle protein degradation through a pathway involving the cAMP cascade. The present work investigated the systemic effect of pentoxifylline (PTX; cAMP-phosphodiesterase inhibitor) treatment on the rate of overall proteolysis, the activity of proteolytic systems, and the process of protein synthesis in extensor digitorum longus muscles from normal and acutely diabetic rats. The direct in vitro effect of this drug on the rates of muscle protein degradation was also investigated. Muscles from diabetic rats treated with PTX showed an increase (22%) in the cAMP content and reduction in total rates of protein breakdown and in activity of Ca2+-dependent (47%) and ATP proteasome dependent (23%) proteolytic pathways. The high content of m-calpain observed in muscles from diabetic rats was abolished by PTX treatment. The addition of PTX (10(-3) M) to the incubation medium increased the cAMP content in muscles from normal (22%) and diabetic (51%) rats and induced a reduction in the rates of overall proteolysis that was accompanied by decreased activity of the Ca2+ dependent and ATP proteasome-dependent proteolytic systems, in both groups. The in vitro addition of H-89, an inhibitor of protein kinase A (PKA), completely blocked the effect of PTX on the reduction of proteolysis in muscles from normal and diabetic rats. The present data suggest that PTX exerts a direct inhibitory effect on protein degradative systems in muscles from acutely diabetic rats, probably involving the participation of cAMP intracellular pathways and activation of PKA, independently of tumor necrosis factor-alpha inhibition. PMID- 17077346 TI - Lactate release from adipose tissue and skeletal muscle in vivo: defective insulin regulation in insulin-resistant obese women. AB - To study the local tissue lactate production in the normal state and its possible disturbances in insulin resistance, rates of lactate release from adipose tissue (AT) and skeletal muscle (SM) were compared postabsorptively and during a hyperinsulinemic euglycemic clamp in 11 healthy nonobese and 11 insulin-resistant obese women. A combination of microdialysis, to measure interstitial lactate, and the 133Xe clearance technique, to determine local blood flow, were used. In the controls, local blood flow increased by 40% in SM (P<0.05) and remained unchanged in AT, whereas the interstitial-plasma difference in lactate doubled in AT (P<0.005) and was unaffected in SM during hyperinsulinemia. In the obese, blood flow and interstitial-plasma difference in lactate remained unchanged in both tissues during hyperinsulinemia. The lactate release (micromol100 g-1min-1) was 1.17+/-0.22 in SM and 0.43+/-0.11 in AT among the controls (P<0.01) and 0.86+/ 0.23 in SM and 0.83+/-0.25 in AT among the obese women in the postabsorptive state. During insulin infusion, lactate release in the controls increased to 1.92+/-0.26 in SM (P<0.005) and to 1.14+/-0.22 in AT (P<0.005) but remained unchanged in the obese women. It is concluded that AT and SM are both significant sources of lactate release postabsorptively, and AT is at least as responsive to insulin as SM. The ability to increase lactate release in response to insulin is impaired in AT and SM in insulin-resistant obese women, involving defective insulin regulation of both tissue lactate metabolism and local blood flow. PMID- 17077347 TI - Prostate-specific antigen and prostate cancer prognosis. PMID- 17077348 TI - Childhood cancer--treatment at a cost. PMID- 17077349 TI - Are Bayes' days upon us? Statistical methods could change the conduct of clinical trials. PMID- 17077350 TI - Stat bite: Number of cancer survivors by site, 2003. PMID- 17077351 TI - Living well post-cancer: care planning for survivors. PMID- 17077352 TI - Geriatric oncology aims for clinical trials. PMID- 17077353 TI - Negative data from lung cancer trial may change practice guidelines, study designs. PMID- 17077354 TI - Detection of life-threatening prostate cancer with prostate-specific antigen velocity during a window of curability. AB - BACKGROUND: Prostate-specific antigen (PSA) level is typically used as a dichotomous test for prostate cancer, resulting in overdiagnosis for a substantial number of men. The rate at which serum PSA levels change (PSA velocity) may be an important indicator of the presence of life-threatening disease. METHODS: PSA velocity was determined in 980 men (856 without prostate cancer, 104 with prostate cancer who were alive or died of another cause, and 20 who died of prostate cancer) who were participants in the Baltimore Longitudinal Study of Aging for up to 39 years. The relative risks (RRs) of prostate cancer death and prostate cancer-specific survival stratified by PSA velocity were evaluated in the three groups of men by Cox regression and Kaplan-Meier analyses. Statistical tests were two-sided. RESULTS: PSA velocity measured 10-15 years before diagnosis (when most men had PSA levels below 4.0 ng/mL) was associated with cancer-specific survival 25 years later; survival was 92% (95% confidence interval [CI] = 84% to 96%) among men with PSA velocity of 0.35 ng/mL per year or less and 54% (95% CI = 15% to 82%) among men with PSA velocity above 0.35 ng/mL per year (P<.001). Furthermore, men with PSA velocity above 0.35 ng/mL per year had a higher relative risk of prostate cancer death than men with PSA velocity of 0.35 ng/mL per year or less (RR = 4.7, 95% CI = 1.3 to 16.5; P = .02); the rates per 100,000 person-years were 1240 for men with a PSA velocity above 0.35 ng/mL per year and 140 for men with a PSA velocity of 0.35 ng/mL per year or less. CONCLUSIONS: PSA velocity may help identify men with life-threatening prostate cancer during a period when their PSA levels are associated with the presence of curable disease. PMID- 17077355 TI - New primary neoplasms of the central nervous system in survivors of childhood cancer: a report from the Childhood Cancer Survivor Study. AB - BACKGROUND: Subsequent primary neoplasms of the central nervous system (CNS) have frequently been described as late events following childhood leukemia and brain tumors. However, the details of the dose-response relationships, the expression of excess risk over time, and the modifying effects of other host and treatment factors have not been well defined. METHODS: Subsequent primary neoplasms of the CNS occurring within a cohort of 14,361 5-year survivors of childhood cancers were ascertained. Each patient was matched with four control subjects by age, sex, and time since original cancer diagnosis. Tumor site-specific radiation dosimetry was performed, and chemotherapy information was abstracted from medical records. Conditional logistic regression was used to estimate odds ratios (ORs), to calculate 95% confidence intervals (CIs), and to model the excess relative risk (ERR) as a function of radiation dose and host factors. For subsequent gliomas, standardized incidence ratios (SIRs) and excess absolute risks (EARs) were calculated based on Surveillance, Epidemiology, and End Results data. RESULTS: Subsequent CNS primary neoplasms were identified in 116 individuals. Gliomas (n = 40) occurred a median of 9 years from original diagnosis; for meningiomas (n = 66), it was 17 years. Radiation exposure was associated with increased risk of subsequent glioma (OR = 6.78, 95% CI = 1.54 to 29.7) and meningioma (OR = 9.94, 95% CI = 2.17 to 45.6). The dose response for the excess relative risk was linear (for glioma, slope = 0.33 [95% CI = 0.07 to 1.71] per Gy, and for meningioma, slope = 1.06 [95% CI = 0.21 to 8.15] per Gy). For glioma, the ERR/Gy was highest among children exposed at less than 5 years of age. After adjustment for radiation dose, neither original cancer diagnosis nor chemotherapy was associated with risk. The overall SIR for glioma was 8.7, and the EAR was 19.3 per 10,000 person-years. CONCLUSIONS: Exposure to radiation therapy is the most important risk factor for the development of a new CNS tumor in survivors of childhood cancers. The higher risk of subsequent glioma in children irradiated at a very young age may reflect greater susceptibility of the developing brain to radiation. PMID- 17077356 TI - Statins, fibrates, and melanoma risk: a systematic review and meta-analysis. AB - BACKGROUND: Large randomized, controlled clinical trials of lovastatin and gemfibrozil for heart disease prevention have reported statistically significantly lower melanoma incidences in persons receiving these medications. Results of in vitro animal model and human case-control studies also suggest that statins and fibrates may reduce the risk of melanoma. METHODS: We performed a systematic review of trials that randomly assigned participants to receive statins or fibrates versus an alternative therapy for a minimum of 6 months. Trials were identified by searching five electronic databases and the reference lists of eligible publications. Unpublished data were solicited from trial investigators and pharmaceutical companies. A meta-analysis was performed using a fixed-effects model, and odds ratios (ORs) with 95% confidence intervals (CIs) were calculated to estimate pooled treatment effects. All statistical tests were two-sided. RESULTS: We obtained data on incident melanomas from 20 of 36 qualifying randomized controlled trials (12 statin trials and eight fibrate trials), with a total of 70,820 participants. A total of 127 melanomas occurred among the 39,426 participants in the statin trials (59 among the 19,872 statin group participants and 68 among the 19,554 control group participants). A total of 27 melanomas occurred among the 31,394 participants enrolled in the fibrate trials (seven among the 12,324 fibrate group participants and 20 among the 19,070 control group participants). Overall, incidence of melanoma was not statistically significantly associated with the use of either statins (OR = 0.87, 95% CI = 0.61 to 1.23) or fibrates (OR = 0.45, 95% CI = 0.20 to 1.01). In a subgroup analysis by drug, only lovastatin use (in one trial) was statistically significantly associated with lower incidence of melanoma (OR = 0.52, 95% CI = 0.27 to 0.99). CONCLUSIONS: These findings do not validate the possibility that statins or fibrates prevent melanoma. PMID- 17077357 TI - Effects of intravenously administered recombinant vesicular stomatitis virus (VSV(deltaM51)) on multifocal and invasive gliomas. AB - BACKGROUND: An ideal virus for the treatment of cancer should have effective delivery into multiple sites within the tumor, evade immune responses, produce rapid viral replication, spread within the tumor, and infect multiple tumors. Vesicular stomatitis virus (VSV) has been shown to be an effective oncolytic virus in a variety of tumor models, and mutations in the matrix (M) protein enhance VSV's effectiveness in animal models. METHODS: We evaluated the susceptibility of 14 glioma cell lines to infection and killing by mutant strain VSV(deltaM51), which contains a single-amino acid deletion in the M protein. We also examined the activity and safety of this strain against the U87 and U118 experimental models of human malignant glioma in nude mice and analyzed the distribution of the virus in the brains of U87 tumor-bearing mice using fluorescence labeling. Finally, we examined the effect of VSV(deltaM51) on 15 primary human gliomas cultured from surgical specimens. All statistical tests were two-sided. RESULTS: All 14 glioma cell lines were susceptible to VSV(deltaM51) infection and killing. Intratumoral administration of VSV(deltaM51) produced marked regression of malignant gliomas in nude mice. When administered systemically, live VSV(deltaM51) virus, as compared with dead virus, statistically significantly prolonged survival of mice with unilateral U87 tumors (median survival: 113 versus 46 days, P = .0001) and bilateral U87 tumors (median survival: 73 versus 46 days, P = .0025). VSV(deltaM51) infected multifocal gliomas, invasive glioma cells that migrated beyond the main glioma, and all 15 primary human gliomas. There was no evidence of toxicity. CONCLUSIONS: Systemically delivered VSV(deltaM51) was an effective and safe oncolytic agent against laboratory models of multifocal and invasive malignant gliomas, the most challenging clinical manifestations of this disease. PMID- 17077358 TI - Efficacy and antivascular effects of EphA2 reduction with an agonistic antibody in ovarian cancer. AB - BACKGROUND: EphA2 is an oncoprotein and tyrosine kinase receptor that is overexpressed in ovarian and many other cancers. We investigated the effects of reduced EphA2 levels on tumor growth and the tumor microenvironment in an orthotopic ovarian cancer model. METHODS: The effect of the EphA2-agonistic monoclonal antibody EA5, alone or in combination with paclitaxel, on the growth of ovarian cancer cells (SKOV3ip1, HeyA8, and HeyA8MDR [taxane-platinum resistant]) was determined in vitro and in vivo by immunoblotting, 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, and immunohistochemical analysis. Expression of EphA2 and markers of angiogenesis (CD31, vascular endothelial growth factor [VEGF], and basic fibroblast growth factor), proliferation (proliferating cell nuclear antigen), and endothelial cell apoptosis (CD31-terminal deoxynucleotidyl transferase biotin-deoxyuridine triphosphate nick-end labeling colocalization) and phosphorylation of Src were analyzed by immunoblotting, immunohistochemistry, immunofluorescence, and in situ hybridization in tumors from treated mice. Statistical tests were two-sided. RESULTS: EA5 antibody treatment led to a more than 90% reduction in EphA2 expression in HeyA8 tumors in vivo. In mice bearing orthotopic SKOV3ip1 or HeyA8 tumors, 4 weeks of EA5 treatment resulted in tumors that weighed 31% and 45% less, respectively, than those in control (IgG-treated) mice (95% confidence interval [CI] = -0.09% to 71% and 20% to 70%, P = .27 and .01, respectively). Combination therapy with EA5 and paclitaxel reduced tumor weight by 77% and 80% (95% CI = 63% to 91% and 68% to 91%), respectively, compared with paclitaxel alone and by 92% and 88% (95% CI = 87% to 97% and 80% to 94%), respectively, compared with IgG alone. Combination therapy also reduced the weight of HeyA8MDR tumors by 47% (95% CI = 24% to 72%) compared with paclitaxel. Mice bearing SKOV3ip1 or HeyA8 tumors that were treated with combination therapy survived longer than those treated with paclitaxel alone (median survival = 144 versus 69 days and 46 versus 37 days, respectively). EA5-treated tumors had reduced microvascular density, proliferation, and VEGF protein and mRNA levels, with increased endothelial cell apoptosis. EphA2 was associated with Src, which was rapidly dephosphorylated after EA5 treatment. CONCLUSIONS: EA5 in combination with paclitaxel decreased tumor growth in an orthotopic ovarian cancer mouse model through antiangiogenic mechanisms associated with reduced levels of VEGF and phosphorylated Src. Humanized antibody constructs against EphA2 are worthy of future study. PMID- 17077359 TI - Re-evaluating adjuvant breast cancer trials: assessing hormone receptor status by immunohistochemical versus extraction assays. AB - BACKGROUND: Tumor levels of steroid hormone receptors, a factor used to select adjuvant treatment for early-stage breast cancer, are currently determined with immunohistochemical assays. These assays have a discordance of 10%-30% with previously used extraction assays. We assessed the concordance and predictive value of hormone receptor status as determined by immunohistochemical and extraction assays on specimens from International Breast Cancer Study Group Trials VIII and IX. These trials predominantly used extraction assays and compared adjuvant chemoendocrine therapy with endocrine therapy alone among pre- and postmenopausal patients with lymph node-negative breast cancer. Trial conclusions were that combination therapy provided a benefit to pre- and postmenopausal patients with estrogen receptor (ER)-negative tumors but not to ER positive postmenopausal patients. ER-positive premenopausal patients required further study. METHODS: Tumor specimens from 571 premenopausal and 976 postmenopausal patients on which extraction assays had determined ER and progesterone receptor (PgR) levels before randomization from October 1, 1988, through October 1, 1999, were re-evaluated with an immunohistochemical assay in a central pathology laboratory. The endpoint was disease-free survival. Hazard ratios of recurrence or death for treatment comparisons were estimated with Cox proportional hazards regression models, and discriminatory ability was evaluated with the c index. All statistical tests were two-sided. RESULTS: Concordance of hormone receptor status determined by both assays ranged from 74% (kappa = 0.48) for PgR among postmenopausal patients to 88% (kappa = 0.66) for ER in postmenopausal patients. Hazard ratio estimates were similar for the association between disease-free survival and ER status (among all patients) or PgR status (among postmenopausal patients) as determined by the two methods. However, among premenopausal patients treated with endocrine therapy alone, the discriminatory ability of PgR status as determined by immunohistochemical assay was statistically significantly better (c index = 0.60 versus 0.51; P = .003) than that determined by extraction assay, and so immunohistochemically determined PgR status could predict disease-free survival. CONCLUSIONS: Trial conclusions in which ER status (for all patients) or PgR status (for postmenopausal patients) was determined by immunohistochemical assay supported those determined by extraction assays. However, among premenopausal patients, trial conclusions drawn from PgR status differed--immunohistochemically determined PgR status could predict response to endocrine therapy, unlike that determined by the extraction assay. PMID- 17077360 TI - Re: Completion of therapy by Medicare patients with stage III colon cancer. PMID- 17077361 TI - Re: Response of a KIT-positive extra-abdominal fibromatosis to imatinib mesylate and KIT genetic analysis. PMID- 17077362 TI - The Cover. Saint Jerome with Saint Paula and Saint Eustochium. PMID- 17077363 TI - A piece of my mind. The hospitalist's story. PMID- 17077364 TI - IOM: overhaul drug safety monitoring. PMID- 17077366 TI - Health experts denounce EPA's soot limits. PMID- 17077365 TI - Critics say drug-eluting stents overused. PMID- 17077367 TI - Cilia database offers clues to diseases. PMID- 17077368 TI - Fondaparinux in patients with ST-segment elevation myocardial infarction. PMID- 17077369 TI - Fondaparinux in patients with ST-segment elevation myocardial infarction. PMID- 17077370 TI - Fondaparinux in patients with ST-segment elevation myocardial infarction. PMID- 17077371 TI - Radiosurgery plus whole-brain radiation therapy for brain metastases. PMID- 17077372 TI - Mutations in families at high risk for breast cancer. PMID- 17077374 TI - Effect of constraint-induced movement therapy on upper extremity function 3 to 9 months after stroke: the EXCITE randomized clinical trial. AB - CONTEXT: Single-site studies suggest that a 2-week program of constraint-induced movement therapy (CIMT) for patients more than 1 year after stroke who maintain some hand and wrist movement can improve upper extremity function that persists for at least 1 year. OBJECTIVE: To compare the effects of a 2-week multisite program of CIMT vs usual and customary care on improvement in upper extremity function among patients who had a first stroke within the previous 3 to 9 months. DESIGN AND SETTING: The Extremity Constraint Induced Therapy Evaluation (EXCITE) trial, a prospective, single-blind, randomized, multisite clinical trial conducted at 7 US academic institutions between January 2001 and January 2003. PARTICIPANTS: Two hundred twenty-two individuals with predominantly ischemic stroke. INTERVENTIONS: Participants were assigned to receive either CIMT (n = 106; wearing a restraining mitt on the less-affected hand while engaging in repetitive task practice and behavioral shaping with the hemiplegic hand) or usual and customary care (n = 116; ranging from no treatment after concluding formal rehabilitation to pharmacologic or physiotherapeutic interventions); patients were stratified by sex, prestroke dominant side, side of stroke, and level of paretic arm function. MAIN OUTCOME MEASURES: The Wolf Motor Function Test (WMFT), a measure of laboratory time and strength-based ability and quality of movement (functional ability), and the Motor Activity Log (MAL), a measure of how well and how often 30 common daily activities are performed. RESULTS: From baseline to 12 months, the CIMT group showed greater improvements than the control group in both the WMFT Performance Time (decrease in mean time from 19.3 seconds to 9.3 seconds [52% reduction] vs from 24.0 seconds to 17.7 seconds [26% reduction]; between-group difference, 34% [95% confidence interval {CI}, 12% 51%]; P<.001) and in the MAL Amount of Use (on a 0-5 scale, increase from 1.21 to 2.13 vs from 1.15 to 1.65; between-group difference, 0.43 [95% CI, 0.05-0.80]; P<.001) and MAL Quality of Movement (on a 0-5 scale, increase from 1.26 to 2.23 vs 1.18 to 1.66; between-group difference, 0.48 [95% CI, 0.13-0.84]; P<.001). The CIMT group achieved a decrease of 19.5 in self-perceived hand function difficulty (Stroke Impact Scale hand domain) vs a decrease of 10.1 for the control group (between-group difference, 9.42 [95% CI, 0.27-18.57]; P=.05). CONCLUSION: Among patients who had a stroke within the previous 3 to 9 months, CIMT produced statistically significant and clinically relevant improvements in arm motor function that persisted for at least 1 year. Trial Registration clinicaltrials.gov Identifier: NCT00057018. PMID- 17077375 TI - Statin therapy and risks for death and hospitalization in chronic heart failure. AB - CONTEXT: Whether statin therapy has beneficial effects on clinical outcomes in patients with heart failure is unclear. OBJECTIVE: To evaluate the association between initiation of statin therapy and risks for death and hospitalization among adults with chronic heart failure. DESIGN, SETTING, AND PATIENTS: Propensity-adjusted cohort study of adults diagnosed with heart failure who were eligible for lipid-lowering therapy but had no previous known statin use, within an integrated health care delivery system in northern California between January 1, 1996, and December 31, 2004. Statin use was estimated from filled outpatient prescriptions in pharmacy databases. MAIN OUTCOME MEASURES: All-cause death and hospitalization for heart failure during a median of 2.4 years of follow-up. We examined the independent relationships between statin therapy and risks for adverse events overall and stratified by the presence or absence of coronary heart disease after multivariable adjustment for potential confounders. RESULTS: Among 24,598 adults diagnosed with heart failure who had no prior statin use, those initiating statin therapy (n = 12,648; 51.4%) were more likely to be younger, male, and have known cardiovascular disease, diabetes, and hypertension. There were 8235 patients who died. Using an intent-to-treat approach, incident statin use was associated with lower risks of death (age- and sex-adjusted rate of 14.5 per 100 person-years with statin therapy vs 25.3 per 100 person-years without statin therapy; adjusted hazard ratio, 0.76 [95% confidence interval, 0.72-0.80]) and hospitalization for heart failure (age- and sex-adjusted rate of 21.9 per 100 person-years with statin therapy vs 31.1 per 100 person-years without statin therapy; adjusted hazard ratio, 0.79 [95% confidence interval, 0.74-0.85]) even after adjustment for the propensity to take statins, cholesterol level, use of other cardiovascular medications, and other potential confounders. Incident statin use was associated with lower adjusted risks of adverse outcomes in patients with or without known coronary heart disease. CONCLUSION: Among adults diagnosed with heart failure who had no prior statin use, incident statin use was independently associated with lower risks of death and hospitalization among patients with or without coronary heart disease. PMID- 17077376 TI - Prevalence of and risk factors for lifetime suicide attempts among blacks in the United States. AB - CONTEXT: Lack of data on the lifetime prevalence and age at onset of suicide ideation, plans, and attempts among blacks in the United States limits the creation and evaluation of interventions to reduce suicide among black Americans. OBJECTIVE: To examine the prevalence and correlates of suicide ideation, planning, and attempts across 2 ethnic classifications of blacks in a nationally representative sample. DESIGN, SETTING, AND PARTICIPANTS: Data are from the National Survey of American Life, a national household probability sample of 5181 black respondents aged 18 years and older, conducted between February 2001 and June 2003, using a slightly modified adaptation of the World Health Organization World Mental Health version of the Composite International Diagnostic Interview. Bivariate and survival analyses were used to delineate patterns and correlates of nonfatal suicidal behavior. MAIN OUTCOME MEASURES: Self-reports of lifetime suicide ideation, planning, and attempts. RESULTS: Survey respondents, categorized as African Americans and Caribbean Americans, reported lifetime prevalence of 11.7% for suicide ideation and 4.1% for attempts. Among the respondents who reported ideation, 34.6% transitioned to making a plan and only 21% made an unplanned attempt. Among 4 ethnic-sex groups, the 7.5% lifetime prevalence for attempts among Caribbean black men was the highest among black Americans. The greatest risk of progressing to suicide planning or attempt among ideators occurred within the first year after ideation onset. Blacks at higher risk for suicide attempts were in younger birth cohorts, less educated, Midwest residents, and had 1 or more Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition disorders. CONCLUSIONS: This study documents the burden of nonfatal suicidality among US blacks, notably Caribbean black men, and individuals making planned attempts. Advancing research on the transition from suicide planning to attempt is vital to the efficacy of health care professionals' ability to screen blacks at risk for suicide. PMID- 17077377 TI - Multiple serotonergic brainstem abnormalities in sudden infant death syndrome. AB - CONTEXT: The serotonergic (5-hydroxytryptamine [5-HT]) neurons in the medulla oblongata project extensively to autonomic and respiratory nuclei in the brainstem and spinal cord and help regulate homeostatic function. Previously, abnormalities in 5-HT receptor binding in the medullae of infants dying from sudden infant death syndrome (SIDS) were identified, suggesting that medullary 5 HT dysfunction may be responsible for a subset of SIDS cases. OBJECTIVE: To investigate cellular defects associated with altered 5-HT receptor binding in the 5-HT pathways of the medulla in SIDS cases. DESIGN, SETTING, AND PARTICIPANTS: Frozen medullae from infants dying from SIDS (cases) or from causes other than SIDS (controls) were obtained from the San Diego Medical Examiner's office between 1997 and 2005. Markers of 5-HT function were compared between SIDS cases and controls, adjusted for postconceptional age and postmortem interval. The number of samples available for each analysis ranged from 16 to 31 for SIDS cases and 6 to 10 for controls. An exploratory analysis of the correlation between markers and 6 recognized risk factors for SIDS was performed. MAIN OUTCOME MEASURES: 5-HT neuron count and density, 5-HT(1A) receptor binding density, and 5 HT transporter (5-HTT) binding density in the medullary 5-HT system; correlation between these markers and 6 recognized risk factors for SIDS. RESULTS: Compared with controls, SIDS cases had a significantly higher 5-HT neuron count (mean [SD], 148.04 [51.96] vs 72.56 [52.36] cells, respectively; P<.001) and 5-HT neuron density (P<.001), as well as a significantly lower density of 5-HT(1A) receptor binding sites (P or =20 weeks' gestation born to US residents. We calculated birthweight-specific infant mortality rates (deaths in each birthweight category per 1000 live births in that category), and overall and cause-specific infant mortality rates (deaths per 100000 live births) in infancy (0-364 days) and in the neonatal (0-27 days) and postneonatal (28-364 days) periods. RESULTS: Birthweight-specific infant mortality rates declined among AIAN and White infants across all birthweight categories, but AIAN infants generally had higher birthweight-specific infant mortality rates. Infant mortality rates declined for both groups, yet in 1998 2000, AIAN infants were still 1.7 times more likely to die than White infants. Most of the disparity was because of elevated post-neonatal mortality, especially from sudden infant death syndrome, accidents, and pneumonia and influenza. CONCLUSIONS: Although birthweight-specific infant mortality rates and infant mortality rates declined among both AIAN and White infants, disparities in infant mortality persist. Preventable causes of infant mortality identified in this analysis should be targeted to reduce excess deaths among AIAN communities. PMID- 17077401 TI - Environmental, social, and personal correlates of having ever had sexual intercourse among American Indian youths. AB - OBJECTIVES: We examined the correlates of having ever had sexual intercourse among American Indians aged 13 to 18 years in Minnesota. METHODS: To assess key environmental, social, and individual correlates of sexual experience, we analyzed data from 4135 American Indian youths who participated in the 1998 and 2001 Minnesota Student Surveys. RESULTS: Forty-two percent of those aged 13 to 15 years and 69% of those aged 16 to 18 years reported that they had ever had sexual intercourse. Correlates of sexual experience varied by age and gender. School connections had the strongest negative associations with sexual experience in young girls, and living with a father had negative associations with sexual experience for younger, but not older, youths. Sexual experience was most strongly and positively associated with risk behaviors such as substance use, violence exposure, and violence perpetuation. CONCLUSIONS: The strongest correlates of sexual experience for American Indian youths were high-risk behaviors and exposure to violence. Future work is needed to develop and employ measures that reflect youth assets and that specifically reflect the experiences of American Indian youths. PMID- 17077402 TI - Social circumstances and education: life course origins of social inequalities in metabolic risk in a prospective national birth cohort. AB - OBJECTIVES: We investigated the relative importance of education and childhood and adult social class in the risk of metabolic syndrome. METHODS: We conducted a prospective birth cohort study of 1311 men and 1318 women aged 53 years in 1999, when metabolic syndrome components were measured. Logistic regression analyses were used to calculate relative index of inequality estimates. RESULTS: Relative to men and women at the highest education levels, men (odds ratio [OR]=2.0; 95% confidence interval [CI]=1.2, 3.2) and women (OR=2.7; 95% CI=1.5, 4.6) with the least education were at twice the risk or more of having the metabolic syndrome. Adjustment for childhood and adult social class strengthened this result among men and weakened it among women. Childhood social class was independently associated with the metabolic syndrome in women (OR=2.0; 95% CI=1.1, 3.6) but not in men (OR=1.1; 95% CI= 0.7, 1.8). Associations between adult social class and the metabolic syndrome or its components were largely accounted for by childhood socioeconomic measures. CONCLUSIONS: Educational differences should be considered in the design of interventions aimed at reducing the burden of the metabolic syndrome in socially disadvantaged groups. PMID- 17077403 TI - Health and health care for the 21st century: for all the people. PMID- 17077404 TI - Beginning to begin: reports from the battle on obesity. PMID- 17077405 TI - Effect of televised, tobacco company-funded smoking prevention advertising on youth smoking-related beliefs, intentions, and behavior. AB - OBJECTIVE: To relate exposure to televised youth smoking prevention advertising to youths' smoking beliefs, intentions, and behaviors. METHODS: We obtained commercial television ratings data from 75 US media markets to determine the average youth exposure to tobacco company youth-targeted and parent-targeted smoking prevention advertising. We merged these data with nationally representative school-based survey data (n = 103,172) gathered from 1999 to 2002. Multivariate regression models controlled for individual, geographic, and tobacco policy factors, and other televised antitobacco advertising. RESULTS: There was little relation between exposure to tobacco company-sponsored, youth-targeted advertising and youth smoking outcomes. Among youths in grades 10 and 12, during the 4 months leading up to survey administration, each additional viewing of a tobacco company parent-targeted advertisement was, on average, associated with lower perceived harm of smoking (odds ratio [OR]=0.93; confidence interval [CI]=0.88, 0.98), stronger approval of smoking (OR=1.11; CI=1.03,1.20), stronger intentions to smoke in the future (OR=1.12; CI=1.04,1.21), and greater likelihood of having smoked in the past 30 days (OR=1.12; CI=1.04,1.19). CONCLUSIONS: Exposure to tobacco company youth-targeted smoking prevention advertising generally had no beneficial outcomes for youths. Exposure to tobacco company parent-targeted advertising may have harmful effects on youth, especially among youths in grades 10 and 12. PMID- 17077406 TI - Addressing health care disparities and increasing workforce diversity: the next step for the dental, medical, and public health professions. AB - The racial/ethnic composition of our nation is projected to change drastically in the coming decades. It is therefore important that the health professions improve their efforts to provide culturally competent care to all patients. We reviewed literature concerning health care disparities and workforce diversity issues- particularly within the oral health field--and provide a synthesis of recommendations to address these issues. This review is highly relevant to both the medical and public health professions, because they are facing similar disparity and workforce issues. In addition, the recent establishment of relationships between oral health and certain systemic health conditions will elevate oral health promotion and disease prevention as important points of intervention in the quest to improve our nation's public health. PMID- 17077407 TI - Ascertainment of Hispanic ethnicity on California death certificates: implications for the explanation of the Hispanic mortality advantage. AB - OBJECTIVES: We determined the size and correlates of underascertainment of Hispanic ethnicity on California death certificates. METHODS: We used 1999 to 2000 vital registration data. We compared Hispanic ethnicity reported on the death certificate to Hispanic ethnicity derived from birthplace for the foreign born and an algorithm that used first and last name and percentage of Hispanics in the county of residence for the US-born. We validated death certificate nativity by comparing data with that in linked Social Security Administration records. RESULTS: Ethnicity and birthplace information was concordant for foreign born Hispanics, who have mortality rates that are 25% to 30% lower than those of non-Hispanic Whites. Death certificates likely underascertain deaths of US-born Hispanics, particularly at older ages, for persons with more education, and in census tracts with lower percentages of Hispanics. Conservative correction for under-ascertainment eliminates the Hispanic mortality advantage for US-born men. CONCLUSIONS: Hispanic ethnicity is accurately ascertained on the California death certificate for immigrants. Immigrant Hispanics have lower age-adjusted mortality rates than do non-Hispanic Whites. For US-born Hispanics, the mortality advantage compared with non-Hispanic Whites is smaller and may be explained by underreporting of Hispanic ethnicity on the death certificate. PMID- 17077408 TI - Oral and pharyngeal cancer incidence and mortality among Hispanics, 1996-2002: the need for ethnoregional studies in cancer research. AB - OBJECTIVES: We investigated whether oral cavity and pharyngeal cancer (OPC) incidence and mortality statistics among Hispanics in New York State differed from those among Hispanics in the United States as a whole. METHODS: OPC incidence and mortality statistics for 1996-2002 were obtained from the New York State Cancer Registry and compared with national statistics released by the Surveillance, Epidemiology, and End Results (SEER) program for the same period. RESULTS: Among Hispanic men, OPC incidence rates were approximately 75% and 89% higher in New York State and New York City, respectively, than national rates reported by the SEER program. No notable differences were identified among Hispanic women. Incidence rates among New York State Hispanic men were 16% higher than those of their non-Hispanic White counterparts. The difference was twice as high (32%) among Hispanic men in New York City. Mortality rates among both men and women exhibited patterns similar to the incidence patterns. CONCLUSIONS: Ethnoregional differences exist in the incidence and mortality rates of OPC in the United States. New York State Hispanic men exhibit much higher incidence and mortality rates than US Hispanics as reported by the SEER program. PMID- 17077409 TI - Racial differences in 30-day mortality for pulmonary embolism. AB - OBJECTIVES: Previous studies reported a higher incidence of in-hospital mortality for Black patients who had pulmonary embolism than for White patients. We used a large statewide database to compare 30-day mortality (defined as death within 30 days from the date of latest hospital admission) for Black and White patients who were hospitalized because of pulmonary embolism. METHODS: The study cohort consisted of 15531 discharged patients who had been treated for pulmonary embolism at 186 Pennsylvania hospitals between January 2000 and November 2002. We used random-effects logistic regression to model 30-day mortality for Black and White patients, and adjusted for patient demographic and clinical characteristics. RESULTS: The unadjusted 30-day mortality rates were 9.0% for White patients, 10.3% for Blacks, and 10.9% for patients of other or unknown race. When adjusted for severity of disease using a validated clinical prognostic model for pulmonary embolism, Black patients had 30% higher odds of 30-day mortality compared with White patients at the same site (adjusted odds ratio = 1.3; 95% confidence interval, 1.1,1.6). Neither insurance status nor hospital volume was a significant predictor of 30-day mortality. CONCLUSION: Black patients who had pulmonary embolism had significantly higher odds of 30-day mortality compared with White patients. PMID- 17077410 TI - Rudolf Carl Virchow: medical scientist, social reformer, role model. PMID- 17077411 TI - Advancing health disparities research within the health care system: a conceptual framework. AB - We provide a framework for health services-related researchers, practitioners, and policy makers to guide future health disparities research in areas ranging from detecting differences in health and health care to understanding the determinants that underlie disparities to ultimately designing interventions that reduce and eliminate these disparities. To do this, we identified potential selection biases and definitions of vulnerable groups when detecting disparities. The key factors to understanding disparities were multilevel determinants of health disparities, including individual beliefs and preferences, effective patient-provider communication; and the organizational culture of the health care system. We encourage interventions that yield generalizable data on their effectiveness and that promote further engagement of communities, providers, and policymakers to ultimately enhance the application and the impact of health disparities research. PMID- 17077414 TI - Economics of central line--associated bloodstream infections. AB - Hospital-acquired infections add considerable morbidity and mortality to patient care. However, a detailed economic analysis of these infections on an individual case basis has been lacking. The authors examined both the hospital revenues and expenses in 54 cases of patients with central line-associated bloodstream infections (CLABs) over 3 years in 2 intensive care units and compared these financial data with patients who were matched for age, severity of illness on admission, and principal diagnosis. The average payment for a case complicated by CLAB was $64 894, and the average expense was $91733 with gross margin of -$26 839 per case and a total loss from operations of $1 449 306 in the 54 cases. The costs of CLABs and the associated complications averaged 43% of the total cost of care. The elimination of these preventable infections constitutes not only an opportunity to improve patient outcomes but also a significant financial opportunity. PMID- 17077412 TI - Residential segregation and injection drug use prevalence among Black adults in US metropolitan areas. AB - OBJECTIVES: We analyzed the relations of two 1990 dimensions of racial residential segregation (isolation and concentration) with 1998 injection drug use prevalence among Black adult residents of 93 large US metropolitan statistical areas (MSAs). METHODS: We estimated injection drug use prevalence among Black adults in each MSA by analyzing 3 databases documenting injection drug users' encounters with the health care system. Multiple linear regression methods were used to investigate the relationship of isolation and concentration to the natural logarithm of Black adult injection drug use prevalence, controlling for possible confounders. RESULTS: The median injection drug use prevalence was 1983 per 100000 Black adults (interquartile range: 1422 to 2759 per 100000). The median isolation index was 0.48 (range: 0.05 to 0.84): in half the MSAs studied, the average Black resident inhabited a census tract where 48% or more of the residents were Black. The multiple regression model indicates that an increase of 0.50 in the isolation index was associated with a 23% increase in injection drug use prevalence among Black adults. Concentration was unrelated to the outcome. CONCLUSIONS: Residential isolation is positively related to Black injection drug use prevalence in MSAs. Research into the pathways linking isolation to injection drug use is needed. PMID- 17077415 TI - Adverse outcomes from hospital-acquired infection in Pennsylvania cannot be attributed to increased risk on admission. AB - In July 2005, Pennsylvania became the first state in the nation to publicly report statewide data on hospital-acquired infections (HAI). The published research brief revealed that 11 668 hospitalizations with HAI had markedly different mortality rates, lengths of stay (LOS), and charges than cases without HAI did. To avoid a possibly biased comparison, a 5 to 1 propensity-matched cohort study was performed. Nine cohorts (ie, heart failure, chronic obstructive pulmonary disease, respiratory failure, pneumonia, hip fracture, major surgical complications, colonic resection, diabetes, and gastrointestinal bleeding) were examined for differences in mortality, LOS, and hospital charges. Statistically significant increases in mortality, LOS, and charges were found among HAI cases. HAI cases had more than a 4 times higher median charge than nonHAI controls did. Observed differences in mortality, LOS, and charges between HAI and non-HAI cases in Pennsylvania cannot be explained on the basis of increased disease-specific severity at the time of admission. PMID- 17077416 TI - Factors associated with risk of surgical wound infections. AB - Surgical wound infections are the most common hospital-acquired infections among patients who undergo inpatient surgery. Risk of infection is a function of both patient susceptibility and exposure. The authors studied all discharges in Pennsylvania from October 1, 2004, through September 30, 2005, in which a circulatory (n= 65 940), neurological (n= 6706), or orthopedic (n = 107 825) procedure was performed using data from the Pennsylvania Health Care Cost Containment Council. They estimated the impact of patient-specific factors on risk of infection and compared the ability of these factors to predict infections relative to hospital effects. Results suggested that for all 3 types of procedures, patient-specific factors were a significant determinant of risk of surgical wound infection. However, prediction of infection was improved by 23% to 33% when hospital fixed effects were included. Although patient-specific factors had a statistically significant association with risk of infections, much of the risk of surgical wound infections is determined by hospital factors. PMID- 17077417 TI - The Lake Wobegon effect: when all the patients are sicker. PMID- 17077418 TI - Will financial incentives stimulate quality improvement? Reactions from frontline physicians. AB - Pay-for-performance is being applied at the physician level to stimulate improvements in quality of care and cost efficiency; however, little is known about how physicians will respond. We interviewed physicians exposed to a financial incentive program in California to identify possible barriers to the successful application of financial incentives by exploring physicians' opinions of and experiences with pay-for-performance programs. Reasons physicians cited for quality deficiencies included insurance coverage limitations and lack of patient compliance, time, and proper physician oversight. Physicians believe that they play a significant role and have a moderate to high degree of control over quality of care and that it is important to self-monitor. Physicians expressed the need for accurate and timely data, peer comparisons, and more patient time, staff support, and consultations with colleagues to successfully monitor and deliver quality care. Many support increased pay for delivering high-quality care but question measurement accuracy, bonus payment financing, and health plan involvement. PMID- 17077419 TI - Attitudes toward nurse practitioner-led chronic disease management to improve outpatient quality of care. AB - To understand the acceptability for a model of chronic disease management, in which primary care patients see nurse practitioners for structured visits using an evidence-based encounter form, the authors sent a mailed survey to primary care physicians and nurse practitioners. A total of 212 subjects completed the survey, for a total response rate of 53% (physicians, 44%; nurse practitioners, 61%). Most physicians (79.5%) reported that nurse practitioners saw patients in their practice. Most physicians (80.0%) and nurse practitioners (95.7%) believed that the proposed model of care would improve the control of chronic illnesses. In addition, most physicians (73.8%) and nurse practitioners (87.6%) believed that the model of care would be of interest to similar providers. Overall, the high level of support for the model and the presence of nurse practitioners in most physician offices suggests that future studies are warranted to understand how best to implement this. PMID- 17077420 TI - Crossing the quality chasm: the role of information technology departments. AB - Integrating information technology (IT) into medical settings is considered essential to transforming hospitals into 21st-century health care institutions. Yet the role of IT departments in maximizing the effectiveness of information systems is not well understood. This article reports a 3-round Delphi panel of Veterans Administration personnel experienced with provider order entry electronic systems. In round 1, 35 administrative, clinical, and IT personnel answered 10 open-ended questions about IT strategies and structures that best support successful transformation. In round 2, panelists rated item importance and ranked proposed strategies. In round 3, panelists received aggregate feedback and rerated the items. Four domains emerged from round 1: IT organization, IT performance monitoring, user-support activities, and core IT responsibilities (eg, computer security, training). In rounds 2 and 3, IT performance monitoring was rated the most important, closely followed by clinical support. Strategies associated with each domain are identified and discussed. PMID- 17077421 TI - Changing physician practice behavior to measure and improve clinical outcomes. AB - Physician practice behavior often produces poor clinical outcomes in the management of cardiovascular disease risk factors in spite of effective treatments and guidelines. The behavior of 165 physicians in 2 settings (suburban and urban) was studied. After collecting baseline clinical data, including systolic blood pressure and low-density lipoprotein cholesterol, a series of interventions was conducted, including academic detailing. Low-density lipoprotein cholesterol decreased 10.4% in suburban patients with cardiovascular disease in the intervention group (P = .001) and 10.5% in the enhanced intervention group (P = .001). Systolic blood pressure decreased 1.11% (P = .357) in the intervention group and 5.13% in the enhanced intervention group (P < .001). In urban hypertensive patients, systolic blood pressure decreased 5.03% (P = .001) and low-density lipoprotein cholesterol decreased 7.01% (P < .001). Combining urban and suburban data, low-density lipoprotein cholesterol decreased 9.32% (P < .001) and systolic blood pressure decreased 4.00% (P < .001). Providing physicians with their clinical outcomes, reviewing national guidelines, and setting expectations, associated with modest practice systems innovations, can produce significant measurable clinical improvements. PMID- 17077422 TI - Impact of a PDA-based diabetes electronic management system in a primary care office. AB - The objective was to determine if a personal digital assistant (PDA) diabetes management system improves processes and outcomes of diabetes care in a primary care setting. Outcomes and processes of care were compared between and within 2 groups of patients with type II diabetes. The intervention group of 58 patients was managed for 16 months with the aid of a PDA diabetes management system. The control group of 115 patients from the same clinic was managed by 1 of 5 physicians not using the diabetes management system. PDA use improved 2 processes of care: annual number of foot and eye examinations (P < .0001 for both). PDA use did not have significant effect on outcomes of care. This PDA disease management system can improve providers' ability to meet practice guidelines for processes of care. However, like other recent studies, there was no improvement in outcomes of diabetes care. PMID- 17077423 TI - Trends in the pattern of care for lung cancer and their correlation with new clinical evidence: experiences in a university-affiliated medical center. AB - The authors surveyed the pattern of care (POC) of lung cancer (LC) using data on 4565 patients from the cancer registry of their university-affiliated hospital institution for the period of 1991 to 2002. New clinical evidence was retrieved from the citations used in the level 1 recommendations in guidelines and textbooks. Using this evidence, indexes (1-6) were chosen, including stage I to II non-small-cell LC (NSCLC): no adjuvant radiotherapy (ART; I); stage III NSCLC: equivocal ART (II), neoadjuvant chemotherapy (C/T; III), and chemoradiotherapy (CRT; IV); stage 4 NSCLC: C/T (V) and limited-stage small-cell LC: CRT (VI). Odds ratios of these index events in the postevidence period versus preevidence period were calculated to show trends in the POC. Trends in the POC were consistent with new clinical evidence with statistical significance. The age and gender adjustment odds ratio was 2.22 to 7.06 for beneficial indexes (3-6) and 0.12 to 0.4 for detrimental indexes (1-2). PMID- 17077424 TI - Care transitions: a threat and an opportunity for patient safety. PMID- 17077426 TI - The schizophrenia drug-treatment paradox: pharmacological treatment based on best possible evidence may be hardest to practise in high-income countries. AB - Most people with schizophrenia live in low- and middle-income countries in which clinicians/policy makers are not the first targets of marketing. Because it is years after a drug is first launched that the full effects become known with confidence, the evidence upon which to base practice in low- and middle-income countries may be less biased than that in richer nations. PMID- 17077427 TI - Suicidal behaviour in youths with depression treated with new-generation antidepressants: meta-analysis. AB - BACKGROUND: Concern exists that antidepressants can cause suicidality in youths with depression. AIMS: To determine the pooled risk of self-harm and suicidal behaviour from randomised trials of newer antidepressants. METHOD: A meta analysis was carried out to calculate odds ratios for the combined data. RESULTS: Self-harm or suicide-related events occurred in 71 of 1487 (4.8%) of depressed youths treated with antidepressants v. 38 of 1254 (3.0%) of those given placebo (fixed effects odds ratio 1.70, 95% CI 1.13-2.54, P=0.01). There was a trend for individual suicidal thoughts, attempts and self-harm to occur more often in youths taking antidepressants than in those given placebo, but none of these differences was statistically significant. CONCLUSIONS: Antidepressants may cause a small short-term risk of self-harm or suicidal events in children and adolescents with major depressive disorder. PMID- 17077428 TI - Progression of mild cognitive impairment to dementia: a challenge to current thinking. AB - BACKGROUND: Studies of conversion from mild cognitive impairment to dementia suggest a linear progression over time. Conversion rates during lifetime may extend to 80-90%. AIMS: This study examines the time-dependent evolution from mild cognitive impairment to dementia. Current assumptions regarding yearly and lifetime conversion rates are challenged. METHOD: A community sample of 1045 dementia-free individuals aged 75 years and over was examined by neuropsychological testing based on 6 years of observation. RESULTS: Approximately 60-65% of people with mild cognitive impairment develop clinical dementia during their life. Progression from mild cognitive impairment to dementia appears to be time dependent, occurring primarily within the initial 18 months. CONCLUSIONS: Further long-term studies are needed to examine the time dependent evolution from mild cognitive impairment to dementia and to establish age-specific conversion rates during lifetime. PMID- 17077429 TI - Music therapy for in-patients with schizophrenia: exploratory randomised controlled trial. AB - BACKGROUND: Music therapy may provide a means of improving mental health among people with schizophrenia, but its effects in acute psychoses have not been explored. AIMS: To examine the feasibility of a randomised trial of music therapy for inpatients with schizophrenia, and explore its effects on mental health. METHOD: Up to 12 weeks of individual music therapy plus standard care were compared with standard care alone. Masked assessments of mental health, global functioning and satisfaction with care were conducted at 3 months. RESULTS: Of 115 eligible patients 81 (70%) were randomised. Two-thirds of those randomised to music therapy attended at least four sessions (median attendance, eight sessions). Multivariate analysis demonstrated a trend towards improved symptom scores among those randomised to music therapy, especially in general symptoms of schizophrenia. CONCLUSIONS: A randomised trial of music therapy for in-patients with schizophrenia is feasible. The effects and cost-effectiveness of music therapy for acute psychosis should be further explored in an explanatory randomised trial. PMID- 17077430 TI - Factors associated with suicidal ideation in the general population: five-centre analysis from the ODIN study. AB - BACKGROUND: Very few studies have examined the cross-national prevalence of suicidal ideation in the general population or variables associated with it. AIMS: To examine the risk factors for suicidal ideas in the general population. METHOD: As part of a five-country two stage epidemiological study of depressive disorder (the ODIN study) a random sample of over 12 000 people were screened using the Beck Depression Inventory (BDI). There followed detailed analysis of item 9 of the BDI, which measured the severity of suicidal ideation. RESULTS: Age, marriage, concern by others and severity of depressed mood independently increased or decreased the odds of suicidal ideation overall. An interaction between life events and social supports was identified, although this differed between men and women. Only concern by others and severity of depression were independently associated with serious suicidal ideation. The study does not allow for interpretation of the direction of the association. CONCLUSIONS: A number of social, clinical and demographic variables were independently associated with all suicidal ideation and with serious suicidal ideation. Longitudinal studies are required to confirm whether these are risk factors for or the result of suicidal ideation or have some other relationship. PMID- 17077431 TI - Individual- and area-level predictors of self-harm repetition. AB - BACKGROUND: No ecological studies have examined the relationship between area characteristics, individual characteristics and self-harm repetition. AIMS: To investigate the association between area-level factors and incidence and repetition of self-harm, and to identify which area-level factors are independently associated with repetition after adjustment for individual factors. METHOD: Prospective cohort study using the Manchester Self-Harm database. Adults who were resident in Manchester and presented to an emergency department following self-harm between 1997 and 2002 were included (n = 4743). The main outcome measure was repeat self-harm within 6 months of the index episode. RESULTS: Four individual factors (previous self-harm, previous psychiatric treatment, employment status, marital status) and one area-based factor (proportion of individuals who were of White ethnicity) were independently associated with repetition. CONCLUSIONS: Repetition of self-harm may be more strongly related to individual factors than to area characteristics. We need to better understand the processes underlying ecological associations with suicidal behaviour before embarking on area-based interventions. PMID- 17077432 TI - Changes in genetic and environmental influences on depressive symptoms across adolescence and young adulthood. AB - BACKGROUND: Depression rises markedly in adolescence, a time when increased and new genetic influences have been reported. AIMS: To examine'new'and'stable' genetic and environmental factors on depressive symptoms in adolescence and young adulthood. METHOD: A questionnaire survey investigated a sample of twin and sibling pairs at three time points over an approximately 3-year period. Over 1800 twin and sibling pairs reported depressive symptoms at the three time points. Data were analysed using multivariate genetic models. RESULTS: Depressive symptoms at all time points were moderately heritable with substantial non-shared environmental contributions. Wave 1 genetic factors accounted for continuity of symptoms at waves 2 and 3. 'New'genetic effects at wave 2 also influenced wave 3 symptoms. New non-shared environmental influences emerged at each time point. CONCLUSIONS: New genetic and environmental influences may explain age-related increases in depression across development. PMID- 17077433 TI - Outcome and prognostic factors for adolescent female in-patients with anorexia nervosa: 9- to 14-year follow-up. AB - BACKGROUND: Earlier studies have indicated poor long-term outcomes for patients with anorexia nervosa. AIMS: To study health and social outcomes of adolescent in patients with anorexia nervosa in relation to prognostic factors. METHOD: A register study based on based on socio-economic and health data was conducted for a national cohort of female residents in Sweden born between 1968 and 1977, including 748 in-patients with anorexia nervosa. RESULTS: At follow-up 9-14 years after hospital admission, 8.7% of patients with anorexia nervosa had persistent psychiatric health problems demanding hospital care and 21.4% were dependent on society for their main income; the stratified relative risks were 5.8 (95% CI 4.7 7.6) and 2.6 (2.3-3.0) respectively, compared with the general female population. The mortality rate for patients with anorexia nervosa was 1.2% and the stratified risk ratio for maternity was 0.6 (95% CI 0.5-0.7). Long duration of hospital care and psychiatric comorbidity were predictors of persistent psychiatric problems and financial dependency on society. CONCLUSIONS: The outcome in this cohort of adolescent in-patients with anorexia nervosa was considerably better than that reported in previous studies. PMID- 17077434 TI - Comparative effectiveness of second-generation antipsychotics and haloperidol in acute schizophrenia. AB - BACKGROUND: There is little information on the comparative effectiveness of second-generation antipsychotic agents. AIMS: To determine if any of five second generation antipsychotics or haloperidol is more effective in treating acutely ill patients with schizophrenia, schizoaffective disorder or schizophreniform disorder. METHOD: A sample of 327 newly admitted patients were randomised to open label treatment with aripiprazole, haloperidol, olanzapine, quetiapine, risperidone or ziprasidone for a minimum of 3 weeks. Measures of effectiveness were improvement in mental status so that the patient no longer required acute in patient care, and changes in Brief Psychiatric Rating Scale (BPRS) scores. RESULTS: By the first measure, haloperidol (89%), olanzapine (92%) and risperidone (88%) were significantly more effective than aripiprazole (64%), quetiapine (64%) and ziprasidone (64%). Changes in BPRS ratings were not significant among treatments. CONCLUSIONS: Haloperidol, olanzapine and risperidone are superior to aripiprazole, quetiapine and ziprasidone for the acute treatment of psychosis in hospitalised patients with schizophrenia, schizoaffective disorder or schizophreniform disorder. PMID- 17077435 TI - Crisis resolution/home treatment teams and psychiatric admission rates in England. AB - BACKGROUND: Introduction of crisis resolution/home treatment teams has been associated with a reduction in hospital admissions in trials. Between 2001 and 2004 there was a rapid expansion in the numbers of these teams in England. AIMS: To examine whether national implementation of these teams was associated with comparable reductions in admissions. METHOD: Observational study using routine data covering working age adult patients in 229 of the 303 local health areas in England from 1998/9 to 2003/4. RESULTS: Admissions fell generally throughout the period, particularly for younger working age adults. Introduction of crisis resolution teams was associated with greater reductions for older working age women (35-64 years); teams always on call were associated with additional reductions for older men and younger women. By the end of the study admissions had fallen by 10% more in the 34 areas with crisis resolution teams in place since 2001, and by 23% more in the 12 of these on call around the clock than in the 130 areas without such teams by 2003/4. Reductions in bed use were smaller. Introduction of assertive outreach teams was not associated with overall reductions in admissions. CONCLUSIONS: Introduction of crisis resolution teams has been associated with reductions in admissions. PMID- 17077436 TI - Clinical and social determinants of duration of untreated psychosis in the AESOP first-episode psychosis study. AB - BACKGROUND: Despite considerable research investigating the relationship between a long duration of untreated psychosis (DUP) and outcomes, there has been much less considering predictors of a long DUP. AIMS: To investigate the clinical and social determinants of DUP in a large sample of patients with a first episode of psychosis. METHOD: All patients with a first episode of psychosis who made contact with psychiatric services over a 2-year period and were living in defined catchment areas in London and Nottingham, UK were included in the AESOP study. Data relating to clinical and social variables and to DUP were collected from patients, relatives and case notes. RESULTS: An insidious mode of onset was associated with a substantially longer DUP compared with an acute onset, independent of other factors. Unemployment had a similar, if less strong, effect. Conversely, family involvement in help-seeking was independently associated with a shorter duration. There was weak evidence that durations were longer in London than in Nottingham. CONCLUSIONS: These findings suggest that DUP is influenced both by aspects of the early clinical course and by the social context. PMID- 17077437 TI - Sustained attention and executive functions in euthymic young people with bipolar disorder. AB - BACKGROUND: Persistent neuropsychological impairments have been reported in the euthymic phase of bipolar affective disorder. However, the findings have been confounded by multiple episodes, chronic illness and residual mood symptoms. AIMS: To assess sustained attention and executive functioning in euthymic young people with bipolar I disorder who had had no more than two affective episodes. METHOD: Thirty euthymic patients (with illness duration of less than 5 years and no more than two affective episodes) and 30 matched healthy individuals were assessed for sustained attention and executive functioning. RESULTS: The bipolar group (mean age 22.4 years, s.d.=2.52; duration of illness 20.87 months, s.d.=14.72), showed impairment on tasks of attention and executive functioning. Multivariate logistic regression analysis demonstrated that deficits in executive functioning differentiated cases from controls. There was no correlation between residual depressive symptoms and neuropsychological performance. CONCLUSIONS: Deficits in attention and executive functioning were present in young people who had experienced only a few episodes of bipolar disorder, suggesting that the deficits are possibly trait abnormalities. Whether these deficits worsen with progression of illness needs to be examined in longitudinal studies. PMID- 17077438 TI - Klinefelter's syndrome (karyotype 47,XXY) and schizophrenia-spectrum pathology. AB - Klinefelter's syndrome, characterised by a 47,XXY chromosomal pattern, has largely been associated with physical abnormalities. Here, we report high levels of schizophrenia-spectrum pathology in 32 men with this syndrome in comparison with 26 healthy controls. This may have implications for treatment of have implications for treatment of Klinefelter's syndrome and suggests that the X chromosome may be involved in the aetiology of schizophrenia. PMID- 17077439 TI - Modafinil and unconstrained motor activity in schizophrenia: double-blind crossover placebo-controlled trial. AB - Avolition affects quality of life in chronic schizophrenia. We investigated the effect of modafinil upon unconstrained motor activity in 18 male patients. In a randomised crossover design study, wrist-worn actigraphic monitors were used to objectively record motor activity over a 20 h period. Patients' total activity was significantly greater when given the drug. These data suggest that modafinil increases quantifiable motor behaviour in schizophrenia and may have an impact on avolition. PMID- 17077440 TI - Neuropsychological performance at the age of 13 years and adult schizophreniform disorder: prospective birth cohort study. AB - We examined neuropsychological functioning at age 13 years in adolescents who later developed schizophreniform disorder, compared with healthy controls and with adolescents diagnosed as having had a manic episode or depression or anxiety disorder. Participants were from an unselected birth cohort. Attentional, executive and motor impairments at age 13 were found in those who later fulfilled diagnostic criteria for schizophreniform disorder, suggesting that these impairments may be the earliest emerging neuropsychological impairments in schizophrenia-related disorders. PMID- 17077441 TI - The ecological fallacy and the gender ratio of suicide in China. AB - China is the only country in which the suicide rate is higher among women than men. We provide a demographic perspective on the gender differential in suicide in China. This shows that the male/female ratio of suicide increased between 1991 and 2001 and there is reason to believe this trend will continue. Among the population subgroups, only young women living in rural areas had much higher suicide rates than their male counterparts. It is argued that consideration of the gender ratio of suicide in China must take age-, gender- and region-specific suicide patterns and the population structure into account. The increasing urbanisation of China is likely to be associated with more male suicides and we predict that before long the male suicide rate will overtake that of females. PMID- 17077442 TI - Physical contact with child and adolescent patients. PMID- 17077443 TI - Collaborative therapy: framework for mental health. PMID- 17077444 TI - Cognitive impairment in euthymic patients with bipolar disorder. PMID- 17077446 TI - Acute and transient psychotic disorders and puerperal psychosis. PMID- 17077447 TI - Delay in onset of action of antidepressants. PMID- 17077449 TI - Primary agoraphobia as a specific phobia. PMID- 17077452 TI - Toward a national framework for the secondary use of health data: an American Medical Informatics Association White Paper. AB - Secondary use of health data applies personal health information (PHI) for uses outside of direct health care delivery. It includes such activities as analysis, research, quality and safety measurement, public health, payment, provider certification or accreditation, marketing, and other business applications, including strictly commercial activities. Secondary use of health data can enhance health care experiences for individuals, expand knowledge about disease and appropriate treatments, strengthen understanding about effectiveness and efficiency of health care systems, support public health and security goals, and aid businesses in meeting customers' needs. Yet, complex ethical, political, technical, and social issues surround the secondary use of health data. While not new, these issues play increasingly critical and complex roles given current public and private sector activities not only expanding health data volume, but also improving access to data. Lack of coherent policies and standard "good practices" for secondary use of health data impedes efforts to strengthen the U.S. health care system. The nation requires a framework for the secondary use of health data with a robust infrastructure of policies, standards, and best practices. Such a framework can guide and facilitate widespread collection, storage, aggregation, linkage, and transmission of health data. The framework will provide appropriate protections for legitimate secondary use. PMID- 17077454 TI - [The evaluation of bioavailability and pharmacokinetic parameters of a new antimicrobial preparation trichomikocid]. AB - Antimicrobial preparations are not effective enough for the normalisation of vaginal microflora, regeneration of it's mucosal layer and thus there are still conditions Amaintained for recidives or reinfection of the disease. In addition, there is a limited arsenal of combinations of trichomikocidal preparations. It is very important problem of modern medicine and pharmacology to create new drugs, especially of natural origin, to treat urogenital trichomonias and trichomona fungal infections. The purpose of the trial was to study biopenetration and pharmacokinetic parameters of preparation trichomikocid suppositories for vaginal and rectal use. In vitro and in vivo investigations have shown that the level of biological penetration is higher during the use of trichomikocid suppositories for vaginal and rectal use (95% and 96%) than during the oral use (75%). Correlations between the data of in vivo and in vitro investigations has been found. PMID- 17077453 TI - Evaluation and certification of computerized provider order entry systems. AB - Computerized physician order entry (CPOE) is an application that is used to electronically write physician orders either in the hospital or in the outpatient setting. It is used in about 15% of U.S. Hospitals and a smaller percentage of ambulatory clinics. It is linked with clinical decision support, which provides much of the value of implementing it. A number of studies have assessed the impact of CPOE with respect to a variety of parameters, including costs of care, medication safety, use of guidelines or protocols, and other measures of the effectiveness or quality of care. Most of these studies have been undertaken at CPOE exemplar sites with homegrown clinical information systems. With the increasing implementation of commercial CPOE systems in various settings of care has come evidence that some implementation approaches may not achieve previously published results or may actually cause new errors or even harm. This has lead to new initiatives to evaluate CPOE systems, which have been undertaken by both vendors and other groups who evaluate vendors, focused on CPOE vendor capabilities and effective approaches to implementation that can achieve benefits seen in published studies. In addition, an electronic health record (EHR) vendor certification process is ongoing under the province of the Certification Commission for Health Information Technology (CCHIT) (which includes CPOE) that will affect the purchase and use of these applications by hospitals and clinics and their participation in public and private health insurance programs. Large employers have also joined this focus by developing flight simulation tools to evaluate the capabilities of these CPOE systems once implemented, potentially linking the results of such programs to reimbursement through pay for performance programs. The increasing role of CPOE systems in health care has invited much more scrutiny about the effectiveness of these systems in actual practice which has the potential to improve their ultimate performance. PMID- 17077455 TI - [Principles of the immune modulation therapy against papilloma virus infections]. AB - The aim of the study was to investigate the effect of Immudone on local immune responses of cervical mucous in patients with papilloma virus cervicitis. 80 women of reproductive age with chronic cervicitis, 60 women with no signs of infection and 70 bioptates obtained from the base of vaginal section of the cervix. Colposcopic, bacterioscopic, patho-histological and immunohistochemical investigations have been performed. After using of Immudone healing of ectopia was observed in the most of cases. As a result of local application of Immudone in patients with PVI in the tissue of cervix the rate of CD4+ cells was increased, expression of receptors of interleukin-2 was increased, CD4/CD8 ratio also increased, quantity of macrophages and natural killers was increased. Local sIgA synthesis was restored. PMID- 17077456 TI - [Peculiarities of the functional state of hypothalamus-hypophysis structures and endocrine status in patients with normogonadotropic primary-ovarian deficiency]. AB - 47 (aged from 17 to 30) women have been selected after the assay with exogenous estradiol, which proved the intactness of the mechanism of the negative feedback between ovaries and hypophysis. At the absence of genetic determination, significantly high indexes of premorbid are identified (infectious and viral diseases, surgical interventions into the area of pelvis minor, autoimmune processes). Menarche and the formation period are within the age norm limits; menstrual irregularities are characterized by the hypomenstrual syndrome. In none of the cases voluminal processes in the hypothalamic-hypophyseal area were identified. The hormonal profile was characterized by significant (p<0.001) decrease of FSH (follicle stimulating hormone), PRL (prolactin) and E(2) (estradiol). The impulse secretion of gonadotropins (had been studied in ten women) evidenced a normal secretion rhythm of gonadotropin releasing hormone by hypothalamus, while the assay with exogenous luliberin showed normal reactivity of gonadotrophs of the hypophysis to hypothalamic stimulation. PMID- 17077457 TI - [Ultrasound pecularities of various forms of normogonadotropic ovarian deficiency]. AB - As a result of examination of 237 (aged 16-31) women with various forms of gynecological neuroendocrine syndrome 3 groups of patients have been identified with identical ultrasonic characteristics of ovaries. The main complaints by the patients have been misbalance of menstrual periods. In 27 patients "diffuse loss of structure" was identified with vanishing of the borderline between the cortical and medullary substance with impaired folliculogenesis, developed against the autoimmune processes mainly (of the thyroid gland and ovaries). In 73 patients "diminished detailing of the structure of the follicular apparatus" is observed with absence of the development of the dominant follicle at ultrasonic monitoring. In this group of women hyperprolactinemia (in 97.3%) with sub clinical course of hypothyreosis (in 49.5%) was identified. In 137 women "hyperplasia of the stroma of the ovary and uniform hardening with indispensable enlargement of the ovaries with single isometric follicular cavities" and considerable fibrosis of the capsule was identified in the absolute majority against the insulin-resistance and compensatory hyperinsulinemia". PMID- 17077458 TI - [Use of orlistat (xenical) in the treatment of women with obesity and disorders of menstrual cycle]. AB - Obesity poses a serious threat for health, being a risk factor for development of heart diseases, diabetes type II, tumors, and reproductive function failure. The aim of this study is to investigate the effect of orlistat (xenical) on the character of menstrual cycle and some metabolic indicators in women with obesity. 17 patients of reproductive age with I-III degree of obesity were investigated. The visceral type of obesity and disorders of menstrual cycle were observed in all patients: oligomenorrhea was observed in 9 (52.9%), amenorrhea in 4 (23.5%)and metrorrhagia in 4 (23.5 %) patients. All the patients received orlistat (xenical) 120 mg 3 times per day during 6 months. Orlistat (xenical) therapy results in significant reduction of body weight (12.3%), body mass index (13.3%), improvement of lipid and carbohydrates metabolisms. Normalization of hormonal levels was registered. As a result of all this the restoration of menstrual cycle and ovulation is registered. Orlistat (xenical) is effective in the treatment of women with obesity and menstrual cycle disorders. PMID- 17077459 TI - [Regulon in the treatment of hyperandrogenism in teenage girls]. AB - The aim of study was to assess the efficiency of the use of combined oral contraceptive with dezogestrel-regulon in the treatment of hyperandrogenism in teenage girls. 14-18 y.o. girls with signs of hyperandrogenism have been investigated. Investigation included: physical examination, assessment of body mass, hirsute number, ultrasonography, FSH, LH, prolactin, testosterone, 17 OH progesterone in blood. RESULTS: All the patients received Regulon from the 5th up to 21st day of menstrual cycle during 6 months. After the 6 months of treatment the regulation of menstrual cycle have been registered in 91% of patients. Reduction of hirsute number (from 24.1+/-1.0 to 11.0+/-1.2 ), ovarian volume by ultrasound (from 16.7+/-3.0 sm(3) to 7.1+/-0.9 sm(3)), level of LH (from 12.7+/ 2.4 to 8.0+/-1.2 IU/L) and testosterone (from 4.2 +/-0.2 to 2.8+/-0.6 nmol/l) were registered. Regulon is effective for the treatment of hyperandrogenism in teenage girls. PMID- 17077460 TI - [Functional state of kidneys and upper urogenital tract in patients with intraligamentary ovarian cysts in pre and postoperative periods during the treatment with spasmolytics]. AB - 25 patients with different types of intraligamentary ovarian cysts were observed. The main diagnostic method of the patients with intraligamentary cysts was ultrasound Dopplerometry of arteries iliaca interna, uterine and renalis using the following indicators: pulsatory index, resistance index, maximal blood speed. Determination of these indicators was conducted before operation, on 10 and 30 days after operation. In post-operation period we found that in the side free from the tumor's development, all doppleromethric indices were substantially normalised. PMID- 17077461 TI - [Transvaginal hydrolaparoscopy: treatment of ovarian endometriomas-preliminary identification of their dislocation by method of aspiration drainage]. AB - Transvaginal hydrolaparoscopy (THL) is considered as a first-line endoscopic method in the investigation of infertility. During the last period it has played important role in the rehabilitation of the natural fertility. The rapid development of this method in the operative settings is very attractive by it's low-invasiveness. Coagulation of endometrial cysts using THL method is very effective. In this case operation can be performed on an outpatient basis. The detection of small endometriomas that are not seen during THL is not a problem only for fertiloscopy. We offered method detecting the location of ovarian small endometrial cysts, which are not discovered by THL. For this purpose we suggest to perform aspiration drainage of small cysts under ultrasound control before THL. This routine manipulation allows not only the determination of the amount of endometrial cysts, but also leaves the aspiration hole on the ovarian surface which indicates on the localization of endometrial inversion. Thus it helps to determine, without any difficulties, the site of coagulation during THL. PMID- 17077462 TI - [Differential-diagnostic features of clinical development of urogenital mycoplasmosis]. AB - Every year urogenital mycoplasma draws the increasing attention, as serious pathological disease of women of reproductive age. During the urogenital mycoplasmosis, parameters of clinical semiology are expressed more significantly than without this disease. Among women with a reproductive pathology on a background of urogenital mycoplasmosis most often took place pathological discharge with a putrefactive smell, irritation, an itch and burning of genitals. Though frequency of such clinical symptoms as the painful sexual intercourse, pain conceded the pathological discharge. It is established, that during urogenital infections among pregnant women and women with a reproductive pathology the wide scale of specific symptoms are peculiar for the mycoplasmosis. PMID- 17077463 TI - [Radio wave and cryodestruction in the complex therapy of cervical lesions]. AB - We have determined the effectiveness of cryodestruction and radio wave surgery during the treatment of background diseases of cervical lesions. To 52 patients with background diseases of cervical lesions the radio wave surgery has been performed and for 81 patients cryodestruction has been carried out, 65 patients got laser, anti-virus and immunomodulating therapy after cryosurgical treatment. The usage of radio wave surgery and cryodestruction helps to avoid unpleasant postoperative consequences such as pain, edema, infection and postoperative shock from loss of blood, which quite often arise after application of traditional methods of surgical operation. The advantage of cryodestruction and radio wave surgery at treatment of background diseases of cervical lesions in comparison with other invasive methods of treatment of given pathology is the absence of scars and preservation of cervical lesions' reproductive function. During the radio wave surgery operation the depth of destruction of tissues are being controlled by the adjustment of the device's capacity, which in our opinion is an advantage of radio wave method of treatment of cervical lesions' pathology in comparison with the method of cryodestruction. PMID- 17077464 TI - [Clinico-morpho-ultrasonographical characteristics of large uterine leiomyoma in females during late reproductive and premenopausal period]. AB - The transvaginal and abdominal ultrasound examination is an effective method of defining the internal multinodular structure of large uterine leiomyoma. Ultrasonic Dopplerography allows us to evaluate uterine internal pattern and the blood flow in the patients with large uterine leiomyoma. The incidence of leiomyoma was the same in late reproductive and premenopausal uteri, although the average number of leiomyoma and the average size of the largest leiomyoma were higher in the premenopausal women. The correlation of the morphological and preoperational results of the ultrasound examination of 21 female with large uteral leiomyoma has been carried out. It has been found that the echographic picture of the nodules is defined by the combinations of the various components possessing the properties of hypoechogeneity (necrosis and the edema of the fibrous structures). Different variants of leiomyoma clinical course in the patients are of interest. The conformity of the echographic picture of large leiomyoma and their morphological structure were revealed at the examination of all the types have made up 94%. The data obtained allowed us to optimize the complex of the diagnostic investigations necessary for revealing proliferative process. PMID- 17077465 TI - [The characteristics of proliferation activity in squamous metaplasia and reserve cell hyperplasia of uterine cervix]. AB - The aim of our study was to investigate the characteristics of proliferation activity in squamous metaplasia and reserve cell hyperplasia of uterine cervix. Four groups of patients were selected: 1st group--34 reproductive patients with reserve cell hyperplasia, 2nd group--21 reproductive patients with squamous metaplasia, 3rd group--12 postmenopausal patients with reserve cell hyperplasia, 4th group--29 postmenopausal patients with squamous metaplasia. These groups included the patients with and without infections. Cervical biopsies were studied by immunohistochemistry using anti-Ki-67 monoclonal antibody. Ki-67(+) reserve cells were counted in reserve cell hyperplasia. Ki-67(+) squamous cells were counted in squamous metaplasia. The results showed that in both squamous metaplasia and reserve cell hyperplasia the number of Ki-67(+) cells, i.e. the intensity of proliferation is depended on the co-existence of infection, as well as on the age of the patient. These data are important in the complex treatment of cervical pseudoerosions. PMID- 17077466 TI - [The morphological characteristics of cervical squamous metaplasia in reproductive women]. AB - The aim of our study was to identify the morphological characteristics of cervical squamous metaplasia in reproductive women. We have performed the biopsy in patients who had the abnormal transformation zone by colposcopy. Cervical biopsies were studied histologically by HandE. The results showed that cervical squamous metaplasia has different clinical importance in reproductive patients. In reproductive women it is associated with cervical intraepithelial neoplasia. In reproductive women assessment of pseudoerosions has important meaning. PMID- 17077467 TI - [Clinical, hormonal and histological features in ovarian stromal hyperthecosis]. AB - The aim of our study was to investigate clinical, hormonal and histological features of ovarian stromal hyperthecosis We have studied 104 patients with a histological diagnosis of ovarian stromal hyperthecosis and hyperplasia. Analyzing the results we can conclude that clinical features of ovarian stromal hyperthecosis and hyperplasia with polycystic ovarian disease and without it were identical and characterised by virilization syndrome, disorders of menstrual cycle and reproductive function; metabolic and vegeto-vascular disorders; breast and endometrial hyperplasias, high blood level of testosterone . The histological study of ovarian stromal hyperthecosis has showed that superficial part of ovarian cortex is hypocellular and fibrotic, contains scattered follicles with few granulosa cells, luteinized internal theca cells, hyperplasia of deep cortex and medulla, corpus luteus is absent. PMID- 17077468 TI - [The morpho-functional changes of the follicular apparatus and yellow body of the ovary in women of the reproductive age with myoma]. AB - It have been studied the morpho-functional changes of the ovaries in women of reproductive age with proliferative myoma of uteri. Based on the analysis of the result of the investigation on the myomatic knots of the women in the condition of high activity of proliferation of myocytes a very complexes, multifactorial pathogenetic mechanisms of the myoma has been outlined. Switch in the common changes in the dynamic oxidative stress and hyperestrogenemia promotes development of the myoma. PMID- 17077469 TI - [Impaired immunological status due to the urogenital chlamydiosis in women of reproductive age and its correction]. AB - The aim of our research was to assess clinical-immunological properties of disease course and efficiency of complex etiotropic, enzyme-immunomodification therapy in women with urogenital chlamydiosis. Out of examined 818 women chlamydia was found in 276. The diagnoses of chlamydiosis in all women was assessed using the following methods: immuno-enzymatic, immunofluorescent and DNA hybridization. Immune responses were studied in regard of forms and severity of the disease and its dynamics, as well as the regimens of treatments and their results. Vilkoxon-Mann-Witney non-parametric criterion U was used for statistical evaluation of the results. For measuring of identity of the various forms of treatment Spirman's coefficient r was used. Urogenital chlamydiosis is characterised by expressed misbalance of immune response of depressive character, the intensity of which depends upon the severity of genital pathology and obstetric anamnesis as well. Oral form of plaferon--LB enhances the immune competent ability of the organism. The most expressive clinical effect was received using the combination of the antibiotic, enzymes and immuno-correction (95%). PMID- 17077470 TI - [Contrast sono hysterosalpingography in the study of endometrial abnormalities and tubal patency in infertile patients]. AB - The aim of this study was to assess the diagnostic value and the usefulness of sono hysterosalpingography (SHG) in the detection of uterine abnormalities and tubal patency, compared with other diagnostic methods among patient with infertility. In a prospective study, 86 patients in the initial stage of the infertility treatment were examined by SHG using saline NaCl infundibular and Echovist as contrast media. Further status of the tubes and uterine cavity was assessed by the "gold standards", laparoscopy and hysteroscopy. Sensitivity, specificity, positive predictive value and the negative predictive value of SHG using NaCl infundibular for evaluation of the uterine cavity were 98.8%, 92.5%, 91.4% and 92%, respectively. Sensitivity, specificity, positive predictive value and the negative predictive value of SHG for the assessment of the tubal status were 97.6%, 89.2%, 76.3% and 77.4%, respectively. There were no evident complications during or after the procedure. Sono hysterosalpingography is useful in making decisions regarding further procedures for the diagnosis and treatment of infertility. This method is simple, safe and cheap for early assessment of the reproductive status of uterine cavity and fallopian tubes and perspective as a routine, first-line infertility investigation. PMID- 17077471 TI - [The role of ethiopathologic approach in pelvic prolapse surgery]. AB - Prior to examining and describing pelvic organ support defects in females, all clinicians should use optimal pelvic prolapse classification. We proposed our original classification which includes both the prolapse grade and etiopathological causes. In our classification crucial point of descent is isthmus of uterus. Such conception will aid in providing a comprehensive and correct surgical treatment to reduce prolapse recurrence. According to ethiopathologic principles in pelvic prolapse surgery, in our study success was obtained in 97.3%. PMID- 17077472 TI - [Assessment of long follow-up results after surgical treatment of stress urinary incontinence using tvt and pubovaginal short skin flap sling]. AB - The aims of present study were to evaluate the quality of women's life (QoL) after surgical treatment of stress urinary incontinence (SUI) and to compare long follow-up results of two different procedures. Quality of life questionnaire represents a useful and cheap tool in the investigation of outcomes of the surgical treatment of stress incontinence. By providing an objective measure of the impact of results on women's lives, quality of life questionnaires allow more comprehensive understanding of disease severity and its effect on psychosocial function. In addition, they have an increasingly important role in clinical research, allow comparing two and more different methods of treatment. QoL of patients after TVT was significantly higher, then after pubovaginal sling using short skin flap (subjective cure 80% versus 66% after 48 months follow up). These benefits in QoL, coupled with the minimally invasive nature of the procedure, suggest that TVT could become a favored treatment option in women with SUI. PMID- 17077473 TI - [Clinical-morphological parallels of malignant ovarian tumours]. AB - The detection of the malignant ovarian tumor takes place at a later stage because of a symptomless process of the tumor formation at an earlier stage, hence leading to the delayed diagnostics and inefficient treatment. The objective of the research is the development of an efficient treatment of the malignant ovarian tumor patients, finding the most adequate methods and a delineation of the proper tactics for the achievement of the improved long-term results. We have studied the medical cards of the patients checked into A. Gvamichava Gynecological Clinic of the National Oncology Center in 1990-2000. The research has led to the conclusion that the epithelial malignant ovarian tumors outnumber the non-epithelial malignant ovarian tumors by 5 to 2. The most widespread histotype of the epithelial malignant ovarian tumors is the serous carcinoma, most frequently detected among the 50-69 year old patients and diagnosed at the third and the fourth stages. In non-epithelial malignant ovarian tumors, the most widespread histotype is the granule-cellular tumors, most frequently detected in the 50-69 age group, diagnosed at the II and the III stages. In our clinic, the surgical treatment, with a subsequent chemotherapy is preferential. PMID- 17077474 TI - [Comparative analysis of methods for diagnostics of chlamydial infection in women of reproductive age]. AB - The aim of this study was to compare the effectiveness of three methods of diagnostics of chlamydial infection: polymerase chain reaction (PCR), direct immunofluorescence (DFA) and enzyme immunoassay (EIA). We calculated sensitivity, specificity, positive and negative predictive values for each method of diagnostics. Blood, cervical specimens for detection of Chlamydia trachomatis were obtained from 103 women of reproductive age. The results of investigation show that: 1. PCR, which has high sensitivity (92.1+/-3.4%) and specificity (95.0+/-3.4%), is an effective method of diagnostics of chlamydial infection. 2. While the positive results of PCR in cervical specimens are strong evidence of presence of chlamydial infections, the negative results of PCR in cervical specimens do not prove the absence of chlamydial infection in internal genital organs of women with infertility. For the diagnostics of chlamydial infection in women of reproductive age simultaneous use of two methods of diagnostics is recommended. PMID- 17077475 TI - [Risk factors of urogenital chlamydiosis in women of reproductive age]. AB - The aim of this study is to assess the risk factors of urogenital chlamydiosis in women of reproductive age. 489 women in the ages of 18-49 years have been questioned and investigated. 268 (55.4%) of them had urogenital chlamydiosis. The control group was formed with 214 women without chlamydial infection. For the assessment of risk factors the odds ratio was calculated. 85.7% of patients were urban citizens with the following education levels: 55.6% of them held high school diplomas and 28.2% had university degrees. The results show that the risk factors of urogenital chlamydiosis are: age <30; presence of more than one sexual partner; more than 2-3 episodes of past pelvic inflammatory diseases and sexually transmitted infections (syphilis, gonorrhea, bacterial vaginosis, trichomoniasis), presence of following sexually transmitted infections: bacterial vaginosis, gonorrhea, trichomoniasis. The assessment of risk factors could be useful for elaborating the screening programs of urogenital chlamydiosis in women of reproductive age. In case of systematic assessment of risk factors probability for prediction dynamics of urogenital chlamydiosis is increased. This could be used as a basis for the chlamydial infection control program. PMID- 17077476 TI - [Infertility structure in patients operated without effect due to polycystic ovaries]. AB - The aim of the investigations was to study the efficiency of demedullation of both ovaries in the case of polycystic ovary syndrome (POS), determination of infertility structure in patients in case of ineffective operation of POS and determination of additional treatment tactics in these patients. In all 245 patients tests of functional diagnostics were carried out as well as the ultrasound investigation of pelvic organs and adrenal glands, X-ray of skull and sella turcica, hysterosalpingography were performed. In blood serum the basal levels of prolactin, LH, FSH, estradiol, progesterone and cortisol were investigated by RIA. In twenty four-hours urine 17 KS, DHA and 17 OKS levels were determined. In all patients laparoscopy, hysteroscopy, chromopertubation with diagnostic curettage of uterus cavity were performed. According to the data in operated patients without a positive effect the most frequent cause of infertility was anovulation. Therefore, even successful surgical intervention and recovery of ovarian function do not guarantee post operational adhesions, which can cause infertility of tuboperitoneal genesis. Treatment tactics of patients operated due to POS without effect can't be standard and depend on revealed pathology discovered in post operational period. PMID- 17077477 TI - [Female reproductive function and immune status after M. Stark's modified cesarean section]. AB - With the aim of studying the causes of peculiarities of female reproductive function and secondary sterility as a result of 100 cesarean sections by M. Stark's modified method, we have attempted to determine the immune status of organisms of women in post-operational period, in post-operational purulent septic complications and in post-operational process of uterus involution. As a result of hysteroscopy and metrosalpingographic tests it has been established that a secondary sterility after cesarean section carried out according to M. Stark modification is manifested 2.5 times less than after traditional method of cesarean section which widely used nowadays. PMID- 17077478 TI - The copper-containing amine oxidase from Arthrobacter globiformis: refinement at 1.55 and 2.20 A resolution in two crystal forms. AB - Copper-containing amine oxidases are found in all the major kingdoms of life. They catalyse the oxidation of organic amines in the presence of molecular dioxygen to aldehydes and hydrogen peroxide. The catalytic centres contain a Cu atom and a topaquinone cofactor formed autocatalytically from a tyrosine residue in the presence of Cu and molecular oxygen. The structure of the Cu-containing amine oxidase from Arthrobacter globiformis, which was previously refined at 1.8 A resolution in space group C2 with unit-cell parameters a = 157.84, b = 63.24, c = 91.98 A, beta = 112.0 degrees [Wilce et al. (1997), Biochemistry, 36, 16116 16133], has been re-refined with newly recorded data at 1.55 A resolution. The structure has also been solved and refined at 2.2 A resolution in a new crystal form, space group C2, with unit-cell parameters a = 158.04, b = 64.06, c = 69.69 A, beta = 111.7 degrees. PMID- 17077479 TI - Expression, purification, crystallization and structure of human adipocyte lipid binding protein (aP2). AB - Human adipocyte lipid-binding protein (aP2) belongs to a family of intracellular lipid-binding proteins involved in the transport and storage of lipids. Here, the crystal structure of human aP2 with a bound palmitate is described at 1.5 A resolution. Unlike the known crystal structure of murine aP2 in complex with palmitate, this structure shows that the fatty acid is in a folded conformation and that the loop containing Phe57 acts as a lid to regulate ligand binding by excluding solvent exposure to the central binding cavity. PMID- 17077481 TI - Structure of the ligand-binding domain (LBD) of human androgen receptor in complex with a selective modulator LGD2226. AB - The androgen receptor (AR) is a ligand-inducible steroid hormone receptor that mediates androgen action, determining male sexual phenotypes and promoting spermatogenesis. As the androgens play a dominant role in male sexual development and function, steroidal androgen agonists have been used clinically for some years. However, there is a risk of potential side effects and most steroidal androgens cannot be dosed orally, which limits the use of these substances. 1,2 Dihydro-6-N,N-bis(2,2,2-trifluoroethyl)amino-4-trifluoromethyl-2-quinolinone (LGD2226) is a synthetic nonsteroidal ligand and a novel selective AR modulator. The crystal structure of the complex of LGD2226 with the androgen receptor ligand binding domain (AR LBD) at 2.1 A was solved and compared with the structure of the AR LBD-R1881 complex. It is hoped that this will aid in further explaining the selectivity of LGD2226 observed in in vitro and in vivo assays and in developing more selective and effective therapeutic agents. PMID- 17077480 TI - Fortuitous structure determination of 'as-isolated' Escherichia coli bacterioferritin in a novel crystal form. AB - Escherichia coli bacterioferritin was serendipitously crystallized in a novel cubic crystal form and its structure could be determined to 2.5 A resolution despite a high degree of merohedral twinning. This is the first report of crystallographic data on 'as-isolated' E. coli bacterioferritin. The ferroxidase active site contains positive difference density consistent with two metal ions that had co-purified with the protein. X-ray fluorescence studies suggest that the metal composition is different from that of previous structures and is a mix of zinc and native iron ions. The ferroxidase-centre configuration displays a similar flexibility as previously noted for other bacterioferritins. PMID- 17077482 TI - Structures of PmSOD1 and PmSOD2, two superoxide dismutases from the protozoan parasite Perkinsus marinus. AB - Perkinsus marinus, a facultative intracellular parasite of the eastern oyster Crassostrea virginica, is responsible for mass mortalities of oyster populations. P. marinus trophozoites survive and proliferate within oyster hemocytes, invading most tissues and fluids, thus causing a systemic infection that eventually kills the host. The phagocytosis of P. marinus trophozoites lacks a respiratory burst, suggesting that the parasite has mechanisms that actively abrogate the host's oxidative defense responses. One mechanism and the first line of defense against oxidative damage is the dismutation of superoxide radical to molecular oxygen and hydrogen peroxide by superoxide dismutases (SODs). P. marinus possesses two iron cofactored SODs, PmSOD1 and PmSOD2. Here, the crystallization and X-ray structures of both PmSOD1 and PmSOD2 are presented. PMID- 17077483 TI - Structure of the orthorhombic form of human inosine triphosphate pyrophosphatase. AB - The structure of human inosine triphosphate pyrophosphohydrolase (ITPA) has been determined using diffraction data to 1.6 A resolution. ITPA contributes to the accurate replication of DNA by cleansing cellular dNTP pools of mutagenic nucleotide purine analogs such as dITP or dXTP. A similar high-resolution unpublished structure has been deposited in the Protein Data Bank from a monoclinic and pseudo-merohedrally twinned crystal. Here, cocrystallization of ITPA with a molar ratio of XTP appears to have improved the crystals by eliminating twinning and resulted in an orthorhombic space group. However, there was no evidence for bound XTP in the structure. Comparison with substrate-bound NTPase from a thermophilic organism predicts the movement of residues within helix alpha1, the loop before alpha6 and helix alpha7 to cap off the active site when substrate is bound. PMID- 17077484 TI - The nucleotide-binding site of Aquifex aeolicus LpxC. AB - The structure of recombinant Aquifex aeolicus UDP-3-O-acyl-N-acetylglucosamine deacetylase (LpxC) in complex with UDP has been determined to a resolution of 2.2 A. Previous studies have characterized the binding sites of the fatty-acid and sugar moieties of the substrate, UDP-(3-O-hydroxymyristoyl)-N-acetylglucosamine, but not that of the nucleotide. The uracil-binding site is constructed from amino acids that are highly conserved across species. Hydrophobic associations with the Phe155 and Arg250 side chains in combination with hydrogen-bonding interactions with the main chain of Glu154 and the side chains of Tyr151 and Lys227 position the base. The phosphate and ribose groups are directed away from the active site and interact with Arg137, Lys156, Glu186 and Arg250. The orientation of the phosphate-ribose tail is not conducive to catalysis, perhaps owing to the position of an inhibitory Zn(2+). However, based on the position of uracil revealed in this study and on the previously reported complex of LpxC with an inhibitor, a model is proposed for substrate binding. PMID- 17077485 TI - Crystallization and structural analysis of GADPH from Spinacia oleracea in a new form. AB - Two crystalline forms of GADPH (D-glyceraldehyde-3-phosphate dehydrogenase) from Spinacia oleracea were obtained using sitting-drop vapor diffusion. Despite the very low concentration of GADPH in the solutions, two crystalline forms were obtained, one of which was the previously reported C222 space group with unit cell parameters a = 155.3, b = 181.7, c = 107.6 A and the other of which belonged to a new space group I4(1)22, with unit-cell parameters a = b = 120.9, c = 154.5 A. Diffraction data were measured from both native and derivatives, yielding structures at a resolution limit of 3.0 A. Differences at the NAD(+)/NADP(+) binding site seen in these structures compared with the previously reported structure with bound coenzyme suggest that conformational changes associated with pyridine-nucleotide binding may play a role in the regulation of this enzyme. PMID- 17077486 TI - Crystallization, preliminary crystallographic analysis and phasing of the thiosulfate-binding protein SoxY from Chlorobium limicola f. thiosulfatophilum. AB - The 22 kDa SoxYZ protein complex from the green sulfur bacterium Chlorobium limicola f. thiosulfatophilum is a central player in the sulfur-oxidizing (Sox) enzyme system of the organism by activating thiosulfate for oxidation by SoxXA and SoxB. It has been proposed that SoxYZ exists as a heterodimer or heterotetramer, but the properties and role of the individual components of the complex thus far remain unknown. Here, the heterologous expression, purification, and the crystallization of stable tetrameric SoxY are reported. Crystals of SoxY diffract to 2.15 A resolution and belong to space group C222(1), with unit-cell parameters a = 41.22, b = 120.11, c = 95.30 A. MIRAS data from Pt(2+)- and Hg(2+) derivatized SoxY crystals resulted in an interpretable electron-density map at 3 A resolution after density modification. PMID- 17077487 TI - Purification, crystallization and preliminary X-ray analysis of the glucosamine-6 phosphate N-acetyltransferase from human liver. AB - Glucosamine-6-phosphate N-acetyltransferase from human liver, which catalyzes the transfer of an acetyl group from acetyl coenzyme A (AcCoA) to the primary amine of D-glucosamine 6-phosphate to form N-acetyl-D-glucosamine 6-phosphate, was expressed in a soluble form from Escherichia coli strain BL21 (DE3). The protein was purified to homogeneity using Ni(2+)-chelating chromatography followed by size-exclusion chromatography. Crystals of the protein were obtained by the hanging-drop vapour-diffusion method and diffracted to 2.6 A resolution. The crystals belonged to space group P4(1)2(1)2 or P4(3)2(1)2, with unit-cell parameters a = b = 50.08, c = 142.88 A. PMID- 17077488 TI - New crystal forms of Diocleinae lectins in the presence of different dimannosides. AB - Studying the interactions between lectins and sugars is important in order to explain the differences observed in the biological activities presented by the highly similar proteins of the Diocleinae subtribe. Here, the crystallization and preliminary X-ray data of Canavalia gladiata lectin (CGL) and C. maritima lectin (CML) complexed with Man(alpha1-2)Man(alpha1)OMe, Man(alpha1-3)Man(alpha1)OMe and Man(alpha1-4)Man(alpha1)OMe in two crystal forms [the complexes with Man(alpha1 3)Man(alpha1)OMe and Man(alpha1-4)Man(alpha1)OMe crystallized in space group P3(2) and those with Man(alpha1-2)Man(alpha1)OMe crystallized in space group I222], which differed from those of the native proteins (P2(1)2(1)2 for CML and C222 for CGL), are reported. The crystal complexes of ConA-like lectins with Man(alpha1-4)Man(alpha1)OMe are reported here for the first time. PMID- 17077489 TI - Crystallization and preliminary X-ray diffraction analysis of the Bacillus subtilis replication termination protein in complex with the 37-base-pair TerI binding site. AB - The replication terminator protein (RTP) of Bacillus subtilis binds to specific DNA sequences that halt the progression of the replisome in a polar manner. These terminator complexes flank a defined region of the chromosome into which they allow replication forks to enter but not exit. Forcing the fusion of replication forks in a specific zone is thought to allow the coordination of post-replicative processes. The functional terminator complex comprises two homodimers each of 29 kDa bound to overlapping binding sites. A preparation of RTP and a 37-base-pair TerI sequence (comprising two binding sites for RTP) has been purified and crystallized. A data set to 3.9 A resolution with 97.0% completeness and an R(sym) of 12% was collected from a single flash-cooled crystal using synchrotron radiation. The diffraction data are consistent with space group P622, with unit cell parameters a = b = 118.8, c = 142.6 A. PMID- 17077490 TI - Expression, purification, crystallization and preliminary X-ray diffraction analysis of alpha-11 giardin from Giardia lamblia. AB - Alpha-11 Giardin, a protein from the annexin superfamily, is a 35.0 kDa protein from the intestinal protozoan parasite Giardia lamblia which triggers a form of diarrhea called giardiasis. Here, the cloning, expression, purification and the crystallization of alpha-11 giardin under two different conditions and in two different space groups is reported. Crystals from the first condition diffracted to 1.1 A and belong to a primitive orthorhombic space group, while crystals from the second condition, which included calcium in the crystallization solution, diffracted to 2.93 A and belong to a primitive monoclinic space group. Determination of the detailed atomic structure of alpha-11 giardin will provide a better insight into its biological function and might establish whether this class of proteins is a potential drug target against giardiasis. PMID- 17077491 TI - Cloning, crystallization and preliminary X-ray studies of XC2981 from Xanthomonas campestris, a putative CutA1 protein involved in copper-ion homeostasis. AB - Divalent metal ions play key roles in all living organisms, serving as cofactors for many proteins involved in a variety of electron-transfer activities. However, copper ions are highly toxic when an excessive amount is accumulated in a cell. CutA1 is a protein found in all kingdoms of life that is believed to participate in copper-ion tolerance in Escherichia coli, although its specific function remains unknown. Several crystal structures of multimeric CutA1 with different rotation angles and degrees of interaction between trimer interfaces have been reported. Here, the cloning, expression, crystallization and preliminary X-ray analysis of XC2981, a possible CutA1 protein present in the plant pathogen Xanthomonas campestris, are reported. The XC2981 crystals diffracted to a resolution of 2.6 A. They are cubic and belong to space group I23, with unit-cell parameters a = b = c = 130.73 A. PMID- 17077492 TI - Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of DapA (Rv2753c) from Mycobacterium tuberculosis. AB - Dihydrodipicolinate synthase from Mycobacterium tuberculosis (DHDPS, DapA, Rv2753c) has been cloned, heterologously expressed in Escherichia coli, purified using standard chromatographic techniques and crystallized in a monoclinic crystal form. Preliminary diffraction data analysis suggests the presence of two independent tetramers in the asymmetric unit in almost the same relative orientation. PMID- 17077493 TI - Purification and crystallization of the human EF-hand tumour suppressor protein S100A2. AB - S100A2 is a Ca(2+)-binding EF-hand protein that is mainly localized in the nucleus. There, it acts as a tumour suppressor by binding and activating p53. Wild-type S100A2 and a S100A2 variant lacking cysteines have been purified. CD spectroscopy showed that there are no changes in secondary-structure composition. The S100A2 mutant was crystallized in a calcium-free form. The crystals, with dimensions 30 x 30 x 70 microm, diffract to 1.7 A and belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 43.5, b = 57.8, c = 59.8 A, alpha = beta = gamma = 90 degrees. Preliminary analysis of the X-ray data indicates that there are two subunits per asymmetric unit. PMID- 17077494 TI - Crystallization of Ranasmurfin, a blue-coloured protein from Polypedates leucomystax. AB - Ranasmurfin, a previously uncharacterized approximately 13 kDa blue protein found in the nests of the frog Polypedates leucomystax, has been purified and crystallized. The crystals are an intense blue colour and diffract to 1.51 A with P2(1) symmetry and unit-cell parameters a = 40.9, b = 59.9, c = 45.0 A, beta = 93.3 degrees . Self-rotation function analysis indicates the presence of a dimer in the asymmetric unit. Biochemical data suggest that the blue colour of the protein is related to dimer formation. Sequence data for the protein are incomplete, but thus far have identified no model for molecular replacement. A fluorescence scan shows a peak at 9.676 keV, indicating that the protein binds zinc and suggesting a route for structure solution. PMID- 17077495 TI - Expression, crystallization and preliminary X-ray crystallographic analyses of two N-terminal acetyltransferase-related proteins from Thermoplasma acidophilum. AB - N-terminal acetylation is one of the most common protein modifications in eukaryotes, occurring in approximately 80-90% of cytosolic mammalian proteins and about 50% of yeast proteins. ARD1 (arrest-defective protein 1), together with NAT1 (N-acetyltransferase protein 1) and possibly NAT5, is responsible for the NatA activity in Saccharomyces cerevisiae. In mammals, ARD1 is involved in cell proliferation, neuronal development and cancer. Interestingly, it has been reported that mouse ARD1 (mARD1(225)) mediates epsilon-acetylation of hypoxia inducible factor 1alpha (HIF-1alpha) and thereby enhances HIF-1alpha ubiquitination and degradation. Here, the preliminary X-ray crystallographic analyses of two N-terminal acetyltransferase-related proteins encoded by the Ta0058 and Ta1140 genes of Thermoplasma acidophilum are reported. The Ta0058 protein is related to an N-terminal acetyltransferase complex ARD1 subunit, while Ta1140 is a putative N-terminal acetyltransferase-related protein. Ta0058 shows 26% amino-acid sequence identity to both mARD1(225) and human ARD1(235). The sequence identity between Ta0058 and Ta1140 is 28%. Ta0058 and Ta1140 were overexpressed in Escherichia coli fused with an N-terminal purification tag. Ta0058 was crystallized at 297 K using a reservoir solution consisting of 0.1 M sodium acetate pH 4.6, 8%(w/v) polyethylene glycol 4000 and 35%(v/v) glycerol. X ray diffraction data were collected to 2.17 A. The Ta0058 crystals belong to space group P4(1) (or P4(3)), with unit-cell parameters a = b = 49.334, c = 70.384 A, alpha = beta = gamma = 90 degrees. The asymmetric unit contains a monomer, giving a calculated crystal volume per protein weight (V(M)) of 2.13 A(3) Da(-1) and a solvent content of 42.1%. Ta1140 was also crystallized at 297 K using a reservoir solution consisting of 0.1 M trisodium citrate pH 5.6, 20%(v/v) 2-propanol, 20%(w/v) polyethylene glycol 4000 and 0.2 M sodium chloride. X-ray diffraction data were collected to 2.40 A. The Ta1140 crystals belong to space group R3, with hexagonal unit-cell parameters a = b = 75.174, c = 179.607 A, alpha = beta = 90, gamma = 120 degrees. Two monomers are likely to be present in the asymmetric unit, with a V(M) of 2.51 A(3) Da(-1) and a solvent content of 51.0%. PMID- 17077496 TI - Overexpression, crystallization and preliminary X-ray crystallographic analysis of phosphopantetheine adenylyltransferase from Enterococcus faecalis. AB - Phosphopantetheine adenylyltransferase, an essential enzyme in the coenzyme A biosynthetic pathway, catalyzes the reversible transfer of an adenylyl group from ATP to 4'-phosphopantetheine, yielding 3'-dephospho-CoA and pyrophosphate. Enterococcus faecalis PPAT has been overexpressed in Escherichia coli as a fusion with a C-terminal purification tag and crystallized at 297 K using a reservoir solution consisting of 0.1 M sodium HEPES pH 7.5, 0.8 M sodium dihydrogen phosphate and 0.8 M potassium dihydrogen phosphate. X-ray diffraction data were collected to 2.70 A at 100 K. The crystals belong to the primitive tetragonal space group P4(1) (or P4(3)), with unit-cell parameters a = b = 160.81, c = 225.68 A. Four copies of the hexameric molecule are likely to be present in the asymmetric unit, giving a crystal volume per protein weight (V(M)) of 3.08 A(3) Da(-1) and a solvent content of 60.1%. PMID- 17077497 TI - Preliminary crystallographic characterization of PrnB, the second enzyme in the pyrrolnitrin biosynthetic pathway. AB - Pyrrolnitrin is the active ingredient of drugs for the treatment of superficial fungal infections and was used as a lead structure for the development of fludioxonil. It is an effective agent for plant diseases caused by the fungal pathogen Rhizoctonia solani. Pyrrolnitrin is made in four steps, the second of which, catalyzed by PrnB, is a novel chemical rearrangement of 7 chlorotryptophan. PrnB was overproduced in Pseudomonas fluorescens (BL915) and well diffracting crystals were obtained of a triple cysteine-to-serine mutant by sitting-drop vapour diffusion. Crystals grown in the presence of L-7 chlorotryptophan, D-tryptophan and L-tryptophan are reported. Data sets for each are reported with high-resolution limits of 2.0, 1.75 and 1.75 A, respectively. Two crystals (PrnB in the presence of D-tryptophan and L-7-chlorotryptophan) belong to space group C2 with similar unit-cell parameters (a = 68.6, b = 79.5, c = 92.7 A, alpha = gamma = 90.0, beta = 103.8 degrees). Crystals grown in the presence of L-tryptophan belong to space group C222(1) and have unit-cell parameters a = 67.7, b = 80.1, c = 129.5 A. All crystals contain a monomer in the asymmetric unit. PMID- 17077498 TI - Expression, purification and crystallization of a PCI domain from the COP9 signalosome subunit 7 (CSN7). AB - A core fragment of Arabidopsis thaliana COP9 signalosome (CSN) subunit 7 was expressed in Escherichia coli. The protein was purified to homogeneity and screened for crystallization. Crystallization conditions were refined using the sitting-drop vapour-diffusion method. Crystals were obtained using polyethylene glycol 8000 as a precipitant and have a thick rod-like morphology. Their crystallographic symmetry is orthorhombic, space group C222(1), with unit-cell parameters a = 57.2, b = 86.2, c = 72.6 A and a diffraction limit of 2.06 A. PMID- 17077499 TI - Crystallization and initial crystallographic characterization of the Corynebacterium glutamicum nitrilotriacetate monooxygenase component A. AB - Safety and environmental concerns have recently dictated the proper disposal of nitrilotriacetate (NTA). Biodegradation of NTA is initiated by NTA monooxygenase, which is composed of two proteins: component A and component B. The NTA monooxygenase component A protein from Corynebacterium glutamicum was crystallized using the sitting-drop vapour-diffusion method in the presence of ammonium sulfate as the precipitant. X-ray diffraction data were collected to a maximum resolution of 2.5 A on a synchrotron beamline. The crystal belongs to the monoclinic space group C2, with unit-cell parameters a = 111.04, b = 98.51, c = 171.61 A, beta = 101.94 degrees . The asymmetric unit consists of four molecules, corresponding to a packing density of 2.3 A(3) Da(-1). The structure was solved by molecular replacement. Structure refinement is in progress. PMID- 17077500 TI - Crystallization and preliminary X-ray analysis of human S100A13. AB - S100A13 is a member of the S100 family of EF-hand-containing calcium-binding proteins and plays an important role in the secretion of fibroblast growth factor 1 and interleukin 1alpha, two pro-angiogenic factors released by the endoplasmic reticulum/Golgi-independent non-classical secretory pathway. Human S100A13 was heterologously expressed in Escherichia coli, purified and crystallized by the hanging-drop vapour-diffusion method using PEG 3350 as the precipitant. The crystals diffracted X-rays from a synchrotron-radiation source to 1.8 A resolution and the space group was assigned as primitive orthorhombic P2(1)2(1)2(1). PMID- 17077501 TI - Overexpression, crystallization and preliminary X-ray crystallographic analysis of a putative transposase from Thermoplasma acidophilum encoded by the Ta0474 gene. AB - IS200 transposases, originally identified in Salmonella typhimurium LT2, are present in many bacteria and archaea and are distinct from other groups of transposases. To facilitate further structural comparisons among IS200-like transposases, structural analysis has been initiated of a putative transposase from Thermoplasma acidophilum encoded by the Ta0474 gene. Its 137-residue polypeptide shows high levels of sequence similarity to other members of the IS200 transposase family. The protein was overexpressed in intact form in Escherichia coli and crystallized at 297 K using a reservoir solution consisting of 100 mM Na HEPES pH 7.5 and 20%(v/v) ethanol. X-ray diffraction data were collected to 1.78 A. The crystals belong to the monoclinic space group P2(1), with unit-cell parameters a = 65.00, b = 34.07, c = 121.58 A, alpha = 90, beta = 100.20, gamma = 90 degrees. Four monomers, representing two copies of a dimeric molecule, are present in the asymmetric unit, giving a crystal volume per protein weight (V(M)) of 2.02 A(3) Da(-1) and a solvent content of 39.2%. PMID- 17077502 TI - Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the regulator AcrR from Escherichia coli. AB - This paper describes the cloning, expression, purification and preliminary X-ray data analysis of the AcrR regulatory protein. The Escherichia coli AcrR is a member of the TetR family of transcriptional regulators. It regulates the expression of the AcrAB multidrug transporter. Recombinant AcrR with a 6xHis tag at the C-terminus was expressed in E. coli and purified by metal-affinity chromatography. The protein was crystallized using hanging-drop vapor diffusion. X-ray diffraction data were collected from cryocooled crystals at a synchrotron light source. The best crystal diffracted to 2.5 A. The space group was determined to be P3(2), with unit-cell parameters a = b = 46.61, c = 166.16 A. PMID- 17077503 TI - Crystallization and preliminary X-ray analysis of the aromatic prenyltransferase CloQ from the clorobiocin biosynthetic cluster of Streptomyces roseochromogenes. AB - Crystals of recombinant CloQ (subunit MW = 35 626 Da; 324 amino acids), an aromatic prenyltransferase from Streptomyces roseochromogenes, were grown by vapour diffusion. The protein crystallizes in space group I4(1)22, with unit-cell parameters a = b = 135.19, c = 98.13 A. Native data from a single crystal were recorded to a resolution of 2.2 A in-house. Preliminary analysis of these data indicated that the asymmetric unit corresponds to a monomer, giving an estimated solvent content of 60.6%. CloQ is involved in the biosynthesis of the aminocoumarin antibiotic clorobiocin, which targets the essential bacterial enzyme DNA gyrase. PMID- 17077504 TI - Crystallization and preliminary X-ray diffraction analysis of phospholipid-bound Sfh1p, a member of the Saccharomyces cerevisiae Sec14p-like phosphatidylinositol transfer protein family. AB - Sec14p is the major phosphatidylinositol (PtdIns)/phosphatidylcholine (PtdCho) transfer protein in the budding yeast Saccharomyces cerevisiae and is the founding member of a large eukaryotic protein superfamily. This protein catalyzes the exchange of either PtdIns or PtdCho between membrane bilayers in vitro and this exchange reaction requires no external input of energy or of other protein cofactors. Despite the previous elucidation of the crystal structure of a detergent-bound form of Sec14p, the conformational changes that accompany the phospholipid-exchange reaction remain undefined. Moreover, a structural appreciation of how Sec14p or its homologs bind their various phospholipid substrates remains elusive. Here, the purification and crystallization of yeast Sfh1p, the protein most closely related to Sec14p, are reported. A combination of electrospray ionization mass-spectrometry and collision-induced decomposition mass-spectrometry methods indicate that recombinant Sfh1p loads predominantly with phosphatidylethanolamine. Unlike phospholipid-bound forms of Sec14p, this form of Sfh1p crystallizes readily in the absence of detergent. Sfh1p crystals diffract to 2.5 A and belong to the orthorhombic primitive space group P2(1)2(1)2(1), with unit-cell parameters a = 49.40, b = 71.55, c = 98.21 A, alpha = beta = gamma = 90 degrees. One Sfh1p molecule is present in the asymmetric unit (V(M) = 2.5 A(3) Da(-1); V(s) = 50%). Crystallization of a phospholipid-bound Sec14p-like protein is a critical first step in obtaining the first high resolution picture of how proteins of the Sec14p superfamily bind their phospholipid ligands. This information will significantly extend our current understanding of how Sec14p-like proteins catalyze phospholipid exchange. PMID- 17077505 TI - Expression, purification, crystallization and preliminary X-ray diffraction analysis of mitochondrial thioredoxin Trx3 from Saccharomyces cerevisiae. AB - There are three thioredoxin isoforms in the yeast Saccharomyces cerevisiae: two cytosolic/nuclear thioredoxins, Trx1 and Trx2, and one mitochondrial thioredoxin, Trx3. In the present work, S. cerevisiae Trx3 overexpressed in Escherichia coli was purified and crystallized. The Trx3 crystals were obtained by the hanging drop vapour-diffusion method. A data set diffracting to 2.0 A resolution was collected from a single crystal. The crystal belongs to space group P3(1), with unit-cell parameters a = b = 49.57, c = 94.55 A, alpha = beta = 90, gamma = 120 degrees. The asymmetric unit is assumed to contain two subunits of Trx3, with a V(M) value of 2.62 A(3) Da(-1) and a solvent content of 53%. PMID- 17077506 TI - Crystallization and preliminary X-ray diffraction analysis of two N-terminal fragments of the DNA-cleavage domain of topoisomerase IV from Staphylococcus aureus. AB - DNA topoisomerase IV removes undesirable topological features from DNA molecules in order to help maintain chromosome stability. Two constructs of 56 and 59 kDa spanning the DNA-cleavage domain of the A subunit of topoisomerase IV from Staphylococcus aureus (termed GrlA56 and GrlA59) have been crystallized. Crystals were grown at 291 K using the sitting-drop vapour-diffusion technique with PEG 3350 as a precipitant. Preliminary X-ray analysis revealed that GrlA56 crystals belong to space group P2(1), diffract to a resolution of 2.9 A and possess unit cell parameters a = 83.6, b = 171.5, c = 87.8 A, beta = 90.1 degrees, while crystals of GrlA59 belong to space group P2(1)2(1)2, with unit-cell parameters a = 41.5, b = 171.89, c = 87.9 A. These crystals diffract to a resolution of 2.8 A. This is the first report of the crystallization and preliminary X-ray analysis of the DNA-cleavage domain of a topoisomerase IV from a Gram-positive organism. PMID- 17077507 TI - Substrate deacylation mechanisms of serine-beta-lactamases. AB - The substrate deacylation mechanisms of serine-beta-lactamases (classes A, C and D) were investigated by theoretical calculations. The deacylation of class A proceeds via four elementary reactions. The rate-determining process is the tetrahedral intermediate (TI) formation and the activation energy is 24.6 kcal/mol at the DFT level. The deacylation does not proceed only by Glu166, which acts as a general base, but Lys73 also participates in the reaction. The C3 carboxyl group of the substrate reduces the barrier height at the TI formation (substrate-assisted catalysis). In the case of class C, the deacylation consists of two elementary processes. The activation energy of the TI formation has been estimated to be 30.5 kcal/mol. Tyr150Oeta is stabilized in the deprotonated state in the acyl-enzyme complex and works as a general base. This situation can exist due to the interaction with two positively charged side chains of lysine (Lys67 and Lys315). The deacylation of class D also consists of two elementary reaction processes. The activation energy of the TI formation is ca. 30 kcal/mol. It is thought that the side chain of Lys70 is deprotonated and acts as a general base. When Lys70 is carbamylated, the activation energy is reduced to less than 20 kcal/mol. This suggests that the high hydrolysis activity of class D with carbamylated Lys70 is due to the reduction of activation energy for deacylation. From these results, it is concluded that the contribution of the lysine residue adjacent to the serine residue is indispensable for the enzymatic reactions by serine-beta-lactamases. PMID- 17077508 TI - Pathways of chemical degradation of polypeptide antibiotic bacitracin. AB - We described the main pathways of bacitracin (Bc) decomposition, chromatographically set the position of its major degradation products and evaluated microbiological activity of isolated components of Bc and its degradation products. All processes of Bc decomposition under stress and accelerated test conditions were monitored with HPLC, performed mainly on a new type reversed-phase (RP-18e) monolithic silica column (Chromolith) enabling fast separation times and some of them also on conventional HPLC columns. Diode array detection, preparative HPLC and FAB mass spectrometry were used for identification of individual Bc components. We found that the major decomposition mechanism in water solutions of Bc is oxidation, and in alkaline solutions, deamidation. In oxidation process the components B1, B2 and B3 and A are oxidized into their corresponding oxidative products H1, H2, H3 and F respectively by the same mechanism. A detailed study of oxidative degradation products revealed that HPLC separation with an acid mobile phase caused splitting of peaks of components H2, H3 and F into two peaks but the peak of component H1 did not split due to its special structural properties. For the component A we confirmed gradual formation of desamido product through an intermediate. We found oxidative degradation products of Bc to be relatively stable, and desamido degradation products to be rather unstable. The estimation of kinetics of Bc decomposition was presented with a semi-quantitative model. Microbiological activity of individual isolated active components of Bc was established and the negligible antimicrobial activity of the degradation products was confirmed. PMID- 17077509 TI - Cloning and heterologous expression of a Ca2+-activated chloride channel isoform from rat brain. AB - We previously reported the cloning of a calcium-activated chloride channel (CLCA) from rat brain (Biochem. Biophys. Res. Commun., 334, 569-576 (2005)), which we designated rbCLCA1. We further showed that rbCLCA1 is expressed in the central nervous system and peripheral organs, and may be functionally expressed in mammalian HEK293 cells. In the present study, we report the successful cloning of a second CLCA from rat cerebrum (designated rbCLCA2), using reverse transcription PCR (RT-PCR) with primers specific for rbCLCA1. We have begun to clone this cDNA based on the rbCLCA1-like sequence. The full-length rbCLCA2 cDNA, obtained via 5' and 3' rapid amplification of cDNA ends (RACE), is 2900 bp long and encodes a putative polypeptide of 905 amino acids having at least two major transmembrane domains and showing 85.2% identity to rbCLCA1. RT-PCR analysis revealed that, similar to rbCLCA1, rbCLCA2 was predominantly expressed in the rat cerebrum, cerebellum, kidney, stomach, spinal cord, lung and small intestine, but not in the heart, large intestine, liver, orand spleen. Whole-cell patch clamp studies in HEK293 cells transiently co-transfected with expression vectors encoding rbCLCA2 and EGFP allowed us to identify the presence of niflumic acid (a CLCA channel blocker)-sensitive and voltage-dependent chloride currents in cells expressing rbCLCA2 but not EGFP alone. Treatment of these cells with ionomycin, a Ca2+ ionophore, significantly increased the novel currents in cells expressing rbCLCA2 and EGFP, but not those expressing EGFP alone, indicating that activation of the rbCLCA2 current is Ca2+-dependent. In sum, we herein report the cloning of a second member of the rbCLCA family from rat brain and its functional expression in vitro, thus adding to our knowledge of anion channels and facilitating future exploration of brain and other organ physiology. PMID- 17077510 TI - Long-chain fatty acids induce lipid droplet formation in a cultured human hepatocyte in a manner dependent of Acyl-CoA synthetase. AB - Lipid droplets (LDs) are intracellular storage sites of neutral lipids, which accumulate in fatty liver disease. Here, we investigated the effects of fatty acids and glucose on LD formation in a cultured human hepatocyte, HuH7, by adding them to culture media. Fatty acids with carbohydrate chains C12-C18 efficiently induced LDs, but those of C8 and C10 were ineffective. Glucose did not induce LD formation even in the presence of insulin. Oleic acid induced significant increases in cellular neutral lipids, and cell fractionation revealed that most of the newly synthesized neutral lipids were concentrated in LDs together with LD proteins. The LD formation was not abrogated by removal of medium glucose but was significantly inhibited by an ACSL inhibitor, triacsin C. These results demonstrate that long-chain fatty acids contribute to LD formation to a greater extent than glucose, possibly by being taken up into the cells, activated by ACSL, reconstituted into neutral lipids and then stored in LDs. Pregnenolone and lithium did not suppress oleic acid-dependent LD formation, despite previous reports of their ability to inhibit LD formation in macrophages and adipocytes suggesting differences among LD formations in these cells. PMID- 17077511 TI - Oxidized low-density lipoprotein-binding specificity of the Asp-hemolysin-related synthetic peptides from Aspergillus fumigatus. AB - Oxidatively modified low-density lipoprotein (OxLDL) is present in atherosclerotic lesions and has been proposed to play an important role in atherogenesis. In the present study, in order to clarify the structure-binding activity relationship of Asp-hemolysin-related peptides to OxLDL, we investigated the interaction between Asp-hemolysin-related peptides consisting of 4 to 29 amino acid residues and OxLDL. The incubation of OxLDL with each Asp-hemolysin related peptide resulted in the formation of an Asp-hemolysin/OxLDL complex. In particular, the tetrapeptide, YKDG (P-4), bound to OxLDL and inhibited the OxLDL induced macrophage proliferation in a dose-dependent manner. Furthermore, we demonstrated that lysophosphatidylcholine (LysoPC) extracted from OxLDL inhibited the binding of P-21 to OxLDL in a dose-dependent manner and synthetic [14C]LysoPC bound to P-21. We propose here that the YKDG region is one of the important sites for the binding of these peptides to OxLDL, and LysoPC as a typical lipid moiety of OxLDL is attributed to the binding of OxLDL to these peptides. PMID- 17077512 TI - Identification and characteristics of a novel testis-specific gene, Tsc24, in human and mice. AB - Testis-specific genes are essential for the spermatogenesis in mammalian male reproduction. We have identified a novel gene, Tsc24, from the results of the Affymetrix Genechip analysis in the developmental stage of days 4, 9, 18, 35, 54 and 6 months of postnatal Balb/C mouse testis. The scaling signal intensities of Tsc24 in the six stages of mouse testis showed that the expression of Tsc24 was not detected on day 4, 9 or 18 but on day 35, 54 and 6 months. The full cDNA length of mouse Tsc24 was 899 bp, with a 624 bp open reading frame encoding a 207 amino acid protein with a predicted molecular weight of 23.997 kDa. The results of semi-quantitative RT-PCR showed that the expression of mouse Tsc24 can only be detected after the mouse was 35 d old and the expressing level increased gradually from day 35 to 6 months. Of the eight tissues (liver, spleen, heart, lung, brain, kidney, epididymis, and testis) examined in mice, and of the 12 tissues (liver, kidney, muscle, brain, spleen, adipose, lung, heart, epididymis, testis, ovary and uterus) examined in human, Tsc24 was exclusively expressed in the testis, but in none of the other studied tissues. The result of subcellular localization of GFP-Tsc24 fusion protein in Cos-7 cells supports that Tsc24 protein is expressed in nuclear. Our study should be a basis for function characterization of the Tsc24 gene, leading to the elucidation of the molecular events underlying mammalian male reproduction. PMID- 17077513 TI - Functional relationships between Rad18 and WRNIP1 in vertebrate cells. AB - The WRNIP1 protein interacts with WRN, the product of the causative gene for Werner syndrome. Mutation of the Saccharomyces cerevisiae gene MGS1, the yeast counterpart of WRNIP1, confers synthetic lethality with mutation of RAD18. To examine the functional relationship between WRNIP1 and Rad18 in higher eukaryotic cells, we generated WRNIP1-/-/-/RAD18-/- lines from chicken DT40 cells and compared them with single mutant cell lines. Unlike the corresponding yeast mutant, WRNIP1-/-/-/RAD18-/- cells are viable but grow more slowly than single mutants and wild type cells, and they show an additive or synergistic elevation in the frequency of sister chromatid exchanges. As reported, WRNIP1-/-/- cells and RAD18-/- cells are moderately and severely sensitive to camptothecin (CPT), respectively. Unexpectedly, the severe CPT sensitivity of RAD18-/- cells is slightly suppressed by disruption of the WRNIP1 gene. PMID- 17077514 TI - 2-aminophenoxazine-3-one suppresses the growth of mouse malignant melanoma B16 cells transplanted into C57BL/6Cr Slc mice. AB - Since phenoxazine is an essential structure of actinomycin D, which exerts a strong anticancer effect, we examined the anticancer effect of 2-aminophenoxazine 3-one (Phx-3) on mouse malignant melanoma B16 cells in vitro and in vivo. Phx-3 inhibited proliferation of the B16 cells in a dose-dependent manner in vitro. We furthermore studied the in vivo effects of Phx-3 on mouse malignant melanoma B16 cells transplanted in female C57BL/6Cr Slc mice. Treatment with Phx-3 (0.5 mg/kg) completely suppressed the growth of mouse malignant melanoma B16 cells transplanted in mice as compared with the control group. Phx-3 was found to exert few adverse effects, in terms of bodyweight loss, changes in serum levels of blood biochemical parameters such as aspartate transaminase (AST), alanine transaminase (ALT), blood urea nitrogen (BUN) and creatinine, dysfunction of the liver and the kidney examined by pathological methods, piloerection and wasting, when mice were treated with a dose of 0.5 mg/kg. These results suggest that Phx-3 may be used to treat patients affected by malignant melanoma in future. PMID- 17077515 TI - Antidiabetic, antioxidant and antihyperlipidemic status of Heliotropium zeylanicum extract on streptozotocin-induced diabetes in rats. AB - The potential role of the methanolic extract of Heliotropium zeylanicum (BURM.F) LAMK (MEHZ) in the treatment of diabetes along with its antioxidant and antihyperlipidemic effects was studied in streptozotocin-induced diabetic rats. Oral administration of (MEHZ) 150 and 300 mg/kg/d for 14 d significantly decreased the blood glucose level and considerably increased the body weight, food intake, and liquid intake of diabetic-induced rats. MEHZ significantly decreased thiobarbituric acid reactive substances and significantly increased reduced glutathione, superoxide dismutase and catalase in streptozotocin-induced diabetic rats at the end of 14 d of treatment. The study also investigated the antihyperlipidemic potential of MEHZ. The results show that the active fraction of MEHZ is promising for development of a standardized phytomedicine for the treatment of diabetes mellitus. PMID- 17077516 TI - Anti-ulcer effect of tea catechin in rats. AB - Oral administration of tea catechin dose-dependently prevented absolute ethanol induced (50, 100, 200 mg/kg) or restraint plus water immersion stress-induced acute gastric mucosal injury (300, 400 mg/kg) in rats. When the effect of test compound was evaluated on the 15th day after acetic acid injection to rats, repeated oral administration of tea catechin (25, 50, 100 mg/kg twice daily) dose dependently accelerated the healing of acetic acid-induced chronic gastric ulcers. Tea catechin (10(-5)-10(-1) g/100 ml) concentration-dependently scavenged superoxide anions in vitro. Tea catechin (100, 200 mg/kg orally) markedly inhibited the increase in thiobarbituric acid-reactive substances in the injured mucosa of rats treated with 50% ethanol. Tea catechin (50, 100 mg/kg twice orally, daily) markedly inhibited the increase in content of thiobarbituric acid reactive substances in the ulcerated region of acetic acid-induced gastric ulcers on the 7th and 15th days. In addition, at 50, 100 and 200 mg/kg orally, it dose dependently prevented the decrease in gastric mucosal hexosamine content induced by absolute ethanol, although it failed to inhibit the basal gastric acid secretion. These results suggest that tea catechin may primarily protect gastric mucosa from acute gastric mucosal injury and promote the healing of chronic gastric ulcers by its antioxidant activity and gastric mucus-increasing actions. PMID- 17077517 TI - 2-methoxycinnamaldehyde reduces IL-1beta-induced prostaglandin production in rat cerebral endothelial cells. AB - Prostaglandin E2 (PGE2) works as a common final mediator of the febrile. Guizhi Tang, one of the most famous traditional Chinese medical formula used to treat influenza, common cold and other pyretic conditions, was previously reported to reduce the production of PGE 2 in rats. 2-Methoxycinnamaldehyde is a principle compound isolated from Guizhi-Tang. The aim of the present study was to investigate the effects of 2-methoxycinnamaldehyde on PGE2 production of rat cerebral endothelial cells (CECs). 2-Methoxycinnamaldehyde dose-dependently inhibited interleukin (IL)-1beta-induced PGE2 production in CECs with IC50 values of 174 microM. IL-1beta stimulation increased the protein, activity and mRNA expression of cyclooxygenase (COX)-2 but not COX-1. 2-Methoxycinnamaldehyde reduced IL-1beta-induced protein and activity of COX-2, but did not influence the COX-2 mRNA expression. Our results show that prostaglandin production in CECs during stimulated conditions is sensitive to inhibition by 2 methoxycinnamaldehyde and suggest that 2-methoxycinnamaldehyde may reduce COX-2 protein level and activity but not COX-2 mRNA. PMID- 17077518 TI - Effects of benidipine, a long-lasting dihydropyridine-Ca2+ channel blocker, on cerebral blood flow autoregulation in spontaneously hypertensive rats. AB - Chronic hypertension shifts cerebral blood flow (CBF) autoregulation towards higher blood pressure. We examined whether or not benidipine, a long-lasting dihydropyridine calcium channel blocker (CCB), improves the CBF autoregulation in spontaneously hypertensive rats (SHRs). CBF was analyzed by laser-Doppler flowmetry during stepwise hypotension by controlled bleeding. The lower limit of CBF autoregulation was calculated as the mean arterial blood pressure at which CBF decreased by 10% of the baseline. Mean arterial blood pressure and cerebral vascular resistance in SHRs were higher than those in normotensive Wistar rats. Oral administration of benidipine (3 mg/kg) for 8 d lowered the mean arterial blood pressure and cerebral vascular resistance, which were equivalent to the effects of amlodipine (3 mg/kg), another CCB, or candesartan (1 mg/kg), an Angiotensin II type-1 receptor blocker. The lower limit of CBF autoregulation in SHRs (142+/-4 mmHg) was significantly shifted to a higher-pressure level compared with Wistar rats (59+/-2 mmHg). The lower limit of CBF autoregulation was significantly lower in the benidipine-treated group (91+/-4 mmHg) than that in the control SHRs, and similar to that of the amlodipine group (97+/-6 mmHg). Benidipine reduced the lower limit of CBF autoregulation more effectively than candesartan (109+/-4 mmHg). In conclusion, benidipine shifted the limit of CBF autoregulation towards lower blood pressure in SHRs under hypotensive conditions by hemorrhage. These results suggest that benidipine may be useful for the treatment of hypertensive patients with the elderly or cerebrovascular disorders, in whom autoregulation of CBF is impaired. PMID- 17077519 TI - Antibacterial spectrum of plant polyphenols and extracts depending upon hydroxyphenyl structure. AB - The relationship between the structure and antibacterial activity of 22 polyphenols was analyzed by using minimum inhibitory concentration (MIC) as a criterion against 26 species of bacteria which can grow in Mueller-Hinton medium. There was no clear correlation between Gram-staining and bacterial susceptibility to polyphenols, and the extent of the susceptibility was approximately dependent on the species of bacteria. In the same Gram-negative bacteria, the antibacterial activity of the polyphenols against Aeromonas hydrophila, Vibrio parahaemolyticus and Vibrio vulnificus was comparatively strong. On the other hand, the activity against 11 species of the Enterobacteriaceae was comparatively weak, and the activity against six species of aerobic bacteria causing plant disease was moderate. Polyphenols having pyrogallol groups showed strong antibacterial activity, and those with catechol and resorcinol rings showed lower activity. The structure-activity relationship was extended to 26 polyphenol-rich plant extracts which could have potent antibacterial activity suitable for commercial use. PMID- 17077520 TI - Evaluation of the anti-inflammatory, analgesic and antipyretic activities of the natural polyphenol chlorogenic acid. AB - Phenolic compounds are numerous and ubiquitous in the plant kingdom, being particularly present in health-promoting foods. Epidemiological evidences suggest that the consumption of polyphenol-rich foods reduces the incidence of cancer, coronary heart disease and inflammation. Chlorogenic acid (CGA) is one of the most abundant polyphenol compounds in human diet. Data obtained from in vivo and in vitro experiments show that CGA mostly presents antioxidant and anti carcinogenic activities. However, the effects of CGA on the inflammatory reaction and on the related pain and fever processes have been explored less so far. Therefore, this study was designed to evaluate the anti-inflammatory, antinociceptive and antipyretic activities of CGA in rats. In comparison to control, CGA at doses 50 and 100 mg/kg inhibited carrageenin-induced paw edema beginning at the 2nd hour of the experimental procedure. Furthermore, at doses 50 and 100 mg/kg CGA also inhibited the number of flinches in the late phase of formalin-induced pain test. Such activities may be derived from the inhibitory action of CGA in the peripheral synthesis/release of inflammatory mediators involved in these responses. On the other hand, even at the highest tested dose (200 mg/kg), CGA did not inhibit the febrile response induced by lipopolysaccharide (LPS) in rats. Additional experiments are necessary in order to clarify the true target for the anti-inflammatory and analgesic effects of CGA. PMID- 17077521 TI - Anti-inflammatory activity of crude extract and fractions of Nectandra falcifolia leaves. AB - The present study evaluated the effect of the crude extract of the leaves of Nectandra falcifolia (NEES) Castiglioni and its fractions in different experimental models of inflammation (paw edema, pleurisy, and ear edema). Carrageenan-induced edema of the paw and pleurisy were evaluated in Wistar rats (180-220 g), which were treated with different doses of the total extract (250, 500 mg.kg-1). Edema of the ear, induced by croton oil, and determination of myeloperoxidase activity were evaluated in Swiss mice (25-35 g). In this experiment, the crude extract of Nectandra falcifolia (Nf) (1.25, 2.5, 5.0, 7.5 mg) and the hexane, chloroform, ethyl-acetate and hydromethanol fractions (5.0 mg) were applied topically, immediately after application of the oil. The crude extract of Nf (500 mg.kg-1) significantly reduced edema of the paw compared to the control group. Similarly, at doses of 250 and 500 mg.kg-1 it significantly reduced the volume of pleural inflammatory exudate compared to the control animals. However, it did not change the number of migrated cells. At doses of 2.5, 5.0 and 7.5 mg, the crude extract significantly inhibited edema of the ear and the influx of neutrophils. The fractions from Nectandra falcifolia (hexane, chloroform, ethyl acetate and hydromethanol) also inhibited edema of the ear. Taken together, the results demonstrated that the crude extract and its fractions administered to animals orally or topically showed an anti-inflammatory effect. PMID- 17077522 TI - Biological activities of 2alpha-substituted analogues of 1alpha,25 dihydroxyvitamin D3 in transcriptional regulation and human promyelocytic leukemia (HL-60) cell proliferation and differentiation. AB - Biological activities of 2alpha-substituted 1alpha,25-dihydroxyvitamin D3 analogues were evaluated in vitro. Their binding affinity was examined with calf thymus cytosolic vitamin D receptor (VDR) and rat plasma vitamin D-binding protein (DBP). In addition, the transcriptional activity of the analogues was measured using a rat 25-hydroxyvitamin D3-24-hydroxylase gene promoter, a human osteocalcin gene promoter, and VDR-GAL4 system. This study investigated the biological activities of 2alpha-substituted analogues in comparison with 2beta substitued analogues at the molecular level, with regard to the structural differences of alkyl, hydroxyalkyl, hydroxyalkoxy substituents at the 2-position of 1alpha,25-dihydroxyvitamin D3. PMID- 17077523 TI - The anti-inflammatory effect of Gigukjiwhangwhangami through the inhibition of nuclear factor-kappaB activation in the peripheral blood mononuclear cells of patients with cerebral infarction. AB - The Korean genuine medicine "Gigukjiwhangwhangami (GJWGM)" has long been used for various cerebrovascular diseases. However, the exact mechanism that accounts for the anti-inflammatory effect of GJWGM is not completely understood. The aim of the present study is to elucidate how GJWGM modulates the inflammatory reaction in lipopolysaccaride (LPS)-stimulated peripheral mononuclear cells from patients with cerebral infarction. Production of cytokine was measured by the ELISA and RT PCR method. The level of nuclear factor-kappaB (NF-kappaB)/Rel A protein and NF kappaB DNA binding activity were determined by the Western blot analysis and TF EIA method. We showed that GJWGM inhibited the production of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1alpha, IL-6, and IL-8 induced by LPS in dose dependent manner (p<0.05). Maximal inhibition rate of TNF-alpha, IL-1beta, IL-6 and IL-8 production by GJWGM was about 54.34%, 41.37%, 44.04%, and 54.46%, respectively. GJWGM inhibited the TNF-alpha and IL-8 mRNA expression. In addition, we showed that the inhibitory mechanism of GJWGM is through the suppression of NF-kappaB pathway. Our study suggests that an important molecular mechanism by GJWGM reduce inflammation, which may explain its beneficial effect in the regulation of inflammatory reactions. PMID- 17077524 TI - Dykellic acid inhibits cell migration and tube formation by RhoA-GTP expression. AB - Dykellic acid, a novel factor initially identified from the culture broth of Westerdykella multispora F50733, has been shown to inhibit matrix metalloprotease 9 activity, caspase-3 activity, B cell proliferation and LPS-induced IgM production, suggesting that this factor may have anti-cancer effects. In an effort to further address the possible anti-tumoral effects of dykellic acid, we used wound healing, invasion and RhoA-GTP assays to examine the effects of dykellic acid on cell migration, invasion and angiogenesis. Our results revealed that dykellic acid dose-dependently inhibits B16 cell migration and motility, and inhibits HUVEC tube formation. Western blot analysis of the active form of RhoA (RhoA-GTP) showed that dykellic acid treatment decreased the levels of RhoA-GTP. These findings collectively suggest that dykellic acid may have both anti metastatic and anti-angiogenic acitivites, and provides the first evidence for the involvement of RhoA in dykellic acid-induced effects. PMID- 17077525 TI - Schedule-dependent antitumor activity and toxicity of combinations of 5 fluorouracil and cisplatin/carboplatin against L1210 leukemia-bearing mice. AB - The combination of cisplatin (CDDP) and 5-fluorouracil (5-FU) has been reported to show marked therapeutic effects on experimental tumors and human malignancies, such as head and neck cancer. In these clinical studies, CDDP was administered on day 1 and followed by a 5-d continuous infusion of 5-FU. However, it was repeatedly shown that the sequence of 5-FU followed by CDDP is more active and less toxic in tumor-bearing animals. Thus, the optimal administration schedule of CDDP and 5-FU against L1210 murine leukemia was examined and compared with that of the combination of 5-FU and carboplatin (JM-8). The combinations of 5-FU (days 1 to 5) and CDDP, given either on day 1 or on day 5, showed a similar level of antitumor activity and toxicity. On the other hand, the combinations of 5-FU (days 1 to 5) and JM-8 given on day 5 showed significantly higher antitumor activity and rather less toxicity, as compared with the combinations on the reverse sequence. Thus, the treatment sequence of platinum compounds followed by a 5-d continuous infusion of 5-FU in many clinical studies appears to be extremely favorable to CDDP than JM-8. In addition, pathological examinations of died mice showed that accumulation of ascites and pleural effusion was inhibited most effectively by JM-8, given alone or in combination with 5-FU. These results strongly suggest that the combination of 5-FU followed by JM-8 will be expected to show more excellent antitumor activity against human malignancies, and may be especially useful in patients who are unable to tolerate CDDP-related toxicity. PMID- 17077526 TI - Effect of congestive heart failure on mexiletine pharmacokinetics in a Japanese population. AB - OBJECTIVE: The goal of this study was to evaluate the influence of congestive heart failure (CHF) on the clearance of mexiletine. METHODS: The mexiletine clearance/bioavailability (CL/F) ratio was estimated in 584 inpatients receiving mexiletine therapy. The study population consisted of 210 patients with CHF [CHF group; 116 inpatients with New York Heart Association (NYHA) class I-II (group NYHA I-II) CHF and 94 inpatients with NYHA class III-IV (group NYHA III-IV) CHF] and 374 inpatients without CHF (Non-CHF group). Serum levels of mexiletine were determined by high performance liquid chromatography (HPLC). RESULTS: Mexiletine clearance was significantly lower in the CHF group when compared with the Non-CHF group (0.264+/-0.093 vs. 0.393+/-0.082 l/h/kg, mean+/-S.D., p<0.05). Further, the CL/F ratio was 50% lower in group NYHA III-IV when compared with the Non-CHF group, and the CL/F ratio tended to change in inverse proportion to NYHA class. CONCLUSION: CHF status significantly affects mexiletine clearance. Therefore, dose adjustments and careful monitoring are likely required in CHF patients receiving mexiletine. PMID- 17077527 TI - Skin penetration flux and lag-time of steroids across hydrated and dehydrated human skin in vitro. AB - To study the effect of hydration on skin absorption, we investigated penetration across human skin of twelve model chemicals having steroidal structure but different molecular weight and compared the steady-state penetration rate (J) and lag-time (t) across hydration intact skin (Jh and th) with that across dehydrated intact skin (Jd and td). Stratum corneum (SC) thickness of hydrated (52 microm) is 3.3 times that of dehydrated skin (16 microm). Transepidermal water loss (TEWL) of hydrated (7.6+/-2.1 g/m2/h) is twice that of dehydrated skin (3.4+/-1.6 g/m2/h, p<0.05) which are similar to in vivo values, suggesting the SC barrier function was recovered. The ratio of Jh/Jd ranged between 0.7 and 3.6 (average of 1.9). On the other hand, the ratio of th/td was almost constant (average of 0.8). Ratios of Jh/Jd and th/td were independent of MW and Ko/w. In percutaneous absorption experiments in vitro, skin was preserved in culture medium until use and SC might swell during that time. Therefore, we consider the possibility that J and t varied between hydrated and dehydrated skin. We confirmed the difference of J and t between hydrated and dehydrated skin in vitro and now need to define these results under in vivo condition. PMID- 17077528 TI - The use of heat treatment to eliminate drug interactions due to grapefruit juice. AB - Grapefruit juice (GJ) contains components that may increase the bioavailability of drugs; however, approaches to the removal of these components have been little investigated. It is known that furanocoumarin derivatives (FCs), such as bergamottin (BG) and 6',7'-dihydroxybergamottin (DHB) in GJ, induce such drug interactions. In the present study, it was found that the heat treatment of grapefruit juice decreases concentrations of BG and DHB as well as their interactions both in vitro and in vivo. We incubated GJ for 10, 20, 30, 40, 50, and 60 min at 37, 62, 72, and 95 degrees C; FCs in each sample were then measured, using high-performance liquid chromatography (HPLC). The concentrations of BG and DHB were decreased in a time- and temperature-dependent manner, by 82.5 and 97.9% respectively, after incubation for 1 h at 95 degrees C. In contrast, the concentration of bergaptrol (BT) increased in a time- and temperature dependent manner (27.7% after 60 min at 95 degrees C). In addition, the effect of each GJ sample on testosterone 6beta-oxidation in human liver microsomes was observed. The inhibitory effects of GJ heated to 95 degrees C were decreased in a time-dependent manner, as in the case of BG and DHB concentrations. Furthermore, 2 ml of GJ treated for 60 min at 95 degrees C was administered into the rat duodenum. After 30 min, nifedipine (NFP) was administered intraduodenally at a dose of 3 mg/kg body weight. The concentrations of NFP in the plasma samples were determined by HPLC. No significant increase in the AUC of NFP was observed in the rats given heat-treated GJ. These results suggest that the heat treatment of GJ reduces the concentrations of FCs, thus eliminating the potential for drug interactions. PMID- 17077529 TI - Antitumor activity of liposomal naphthoquinone esters isolated from Thai medicinal plant: Rhinacanthus nasutus KURZ. AB - We previously observed that rhinacanthins-C, -N and -Q, three main naphthoquinone esters isolated from the roots of Thai medicinal plant; Rhinacanthus nasutus KURZ. (Acanthaceae) induced apoptosis of human cervical carcinoma HeLaS3 cells. Since these rhinacanthins showed limited solubility in aqueous medium, we attempted to entrap them into liposomal membrane: Liposomalization enabled injection of the drugs and the drugs were expected to transfer to lipoproteins in the bloodstream. Liposomal formulations of rhinacanthins-C, -N and -Q showed strong antiproliferative activity against HeLaS3 cells with the IC50 values of 32, 17, 70 microM; 19, 17, 52 microM and 2.7, 2.0 and 5.0 microM for the exposure time of 24, 48, and 72 h, respectively. These liposomes suppressed the tumor growth in Meth-A sarcoma-bearing BALB/c mice at the dose of 5.0 mg/kg/d for 10 d. Among rhinacanthins, liposomal rhinacanthin-N significantly suppressed solid tumor growth. Based on these results, our findings demonstrated that rhinacanthin N suppressed tumor growth in vivo, and suggested that liposomes are useful for preparing injectable formulation of hydrophobic drugs. PMID- 17077530 TI - Protective effects of tea melanin against 2,3,7,8-tetrachlorodibenzo-p-dioxin induced toxicity: antioxidant activity and aryl hydrocarbon receptor suppressive effect. AB - We examined the protective ability of tea melanin against 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD)-induced toxicity in C57BL6J mice. Reduced tea melanin (RTM) and non-reduced tea melanin (NRTM) were incorporated to distinguish anti-oxidant activity from alternative pathways. The mice were given a single oral dose of TCDD (100 microg/kg body weight) and then they were administered daily with NRTM or RTM (40 mg/kg, p.o.) for next 14 d. RTM protected the animals against TCDD-induced lipid peroxidation, inhibition of glutathione peroxidase, alteration in reduced and oxidized glutathione concentrations, loss of body weight, and increased relative liver weight. NRTM was less effective as compared to RTM because of its inferior antioxidant activity, but it still displayed a strong protective effect against TCDD toxicity owing to its similar suppression of the activity of the aryl hydrocarbon receptor. Both NRTM and RTM suppressed the expression of CYP1A1 gene and prevented the activation of cytochrome P450 isozyme in the livers of animals exposed to TCDD. These results suggest that tea melanin might be a potential agent offering dual protection against the development of TCDD-induced oxidative stress. PMID- 17077531 TI - Antimicrobial effect of trans-cinnamaldehyde, (-)-perillaldehyde, (-) citronellal, citral, eugenol and carvacrol on airborne microbes using an airwasher. AB - Citral, trans-cinnamaldehyde, (-)-perillaldehyde, (-)-citronellal, eugenol and carvacrol were tested for their influence on microbial count in air by vaporizing with an air washer. The highest antibacterial activity was observed when (-) perillaldehyde was sprayed. The average reduce of germ count was 53%. On the other hand, the antimicrobial activity of eugenol was the lowest of these six compounds. The average reduction of germ count was 13%. When water without volatile compounds was sprayed, the colony forming units increased. These results suggest the utility of selected aroma-compounds for the control of bacteria in the room. PMID- 17077532 TI - Pseudomonas aeruginosa alkaline protease can facilitate siderophore-mediated iron uptake via the proteolytic cleavage of transferrins. AB - In order to determine whether Pseudomonas aeruginosa alkaline protease AprA is involved in facilitating siderophore-mediated iron-acquisition from human transferrins, we measured bacterial growth, the production of siderophore and AprA, iron-acquisition from transferrins, and the proteolytic cleavage of transferrins in an alkaline minimal medium (pH 8.3) containing human transferrins as an iron source and compared these on a time scale. The growth of P. aeruginosa was found to be stimulated in proportion to the iron-saturation levels of transferrins. AprA production and the proteolytic cleavage of transferrins began concomitantly with siderophore production from the early growth phase when P. aeruginosa was actively growing and consuming most iron for growth. However, the AprA-free, but siderophore-containing, culture ultrafiltrates could also remove iron from transferrin. These results indicate that alkaline protease AprA can facilitate the siderophore-mediated iron-uptake of P. aeruginosa via the proteolytic cleavage of transferrins. However, the proteolytic cleavage by AprA is not essentially required for iron-acquisition from transferrins. PMID- 17077533 TI - Antioxidant activity of a novel extract from bamboo grass (AHSS) against ischemia reperfusion injury in rat small intestine. AB - Production of free radical species in cells and body tissues is known to cause many pathological disorders. Therefore, free radical scavengers play an important role in the prevention of various human diseases. Bamboo grass, Sasa senanensis, is a native Japanese plant. Sasa has been used for medicine in Japan for many centuries. In this study, we investigated the antioxidative activity of Absolutely Hemicellulose Senanensis (AHSS), a novel extract from Sasa. In the first part of this study, we found that AHSS has antioxidant activities by the assay using superoxide anion-2-methyl-6-methoxyphenylethynylimidazopyrazynone (MPEC) reaction kit. We then confirmed its antioxidative activity using a rat ischemia and subsequent reperfusion (I/R) injury model. Breakdown of the intestinal wall caused by intestinal I/R was attenuated by pretreatment with AHSS. Moreover, AHSS inhibited the production of lipid peroxide by intestinal I/R. AHSS could be an important source of ingredients for use in functional foods and other applications. PMID- 17077534 TI - Antidepressant-like effects of sarsasapogenin from Anemarrhena asphodeloides BUNGE (Liliaceae). AB - The aim of this study was to investigate the effects of sarsasapogenin from Anemarrhena asphodeloides BUNGE (Liliaceae) on the forced swimming test, and the central noradrenergic, dopaminergic and serotonergic activities in mice. Our results showed that sarsasapogenin treatment at 12.5, 25 and 50 mg/kg (p.o.) for 14 d significantly reduced the duration of immobility in the forced swimming test. These doses that affected the immobile response did not affect locomotor activity. In addition, the neurochemical assays showed that sarsasapogenin produced a marked increase of noradrenaline and serotonin levels at 50 mg/kg in both the hypothalamus and the hippocampus. Moreover, sarsasapogenin showed a monoamine oxidase inhibitory activity in the mouse brain. These findings suggest that the antidepressant activity of sarsasapogenin may involve the central monoaminergic neurotransmitter systems. PMID- 17077535 TI - Trypanocidal activity of triterpenes from Arrabidaea triplinervia and derivatives. AB - Ethanol extract from Arrabidaea triplinervia leaves showed in vitro activity (ED100 5.0 mg/ml) against trypomastigotes of Trypanosoma cruzi, the etiologic agent of Chagas; disease. Bioactivity-directed fractionation of this extract led to the isolation of ursolic and oleanolic acids as trypanocidal compounds besides pomolic acid (not tested) and alpinetine (inactive). A series of natural and synthetic derivatives of ursolic and oleanolic acids was simultaneously assayed for structure activity relationships (SAR) studies. Ursolic acid (ED100 0.4 mg/ml) was four times more active than oleanolic acid (ED100 1.6 mg/ml). The presence of free hydroxy and/or carboxy groups is necessary for the trypanocidal activity as could be deduced from the effect of the acetates, methyl ester, and aldehyde derivatives. PMID- 17077536 TI - Antiallergic activity of aqueous extracts and constituents of Taxus yunnanensis. AB - The H2O, H2O/MeOH (1:1) extracts from the wood of Taxus yunnanensis showed a remarkable inhibitory effect on induced histamine release from the human basophilic cell line, KU812. The eleven constituents purified from the wood extracts of Taxus yunnanensis were tested by an in vitro histamine release inhibition assay. Among them, secoisolarciresinol and taxiresinol were found to show inhibitory activities. A new neolignan, 2-[2-hydroxy-5-(3-hydroxypropyl)-3 methoxyphenyl]-1-(4-hydroxy-3-methoxyphenyl)propane-1,3-diol, was isolated from the wood of Taxus yunnanensis. PMID- 17077537 TI - Development and application of RAPD-SCAR marker for identification of Phyllanthus emblica LINN. AB - Correct genotype identification of medicinal plant material remains important for botanical drug industry. Limitations of chemical and morphological approaches for authentication have generated need for newer methods in quality control of botanicals. The present study was carried out to develop DNA based marker for identification of Phyllanthus emblica LINN. A putative marker (1.1 kb) specific for P. emblica was identified by Random Amplified Polymorphic DNA (RAPD) technique. Sequence Characterized Amplified Region (SCAR) marker was developed from the RAPD amplicon. The SCAR marker was found useful for identification of P. emblica in its commercial samples and Triphalachurna, a multi-component Ayurvedic formulation. PMID- 17077538 TI - 8-hydroxydihydrochelerythrine and 8-hydroxydihydrosanguinarine with a potent acetylcholinesterase inhibitory activity from Chelidonium majus L. AB - Ethanol extract of the aerial portion of Chelidonium majus L. inhibited acetylcholinesterase (AChE) activity without a significant inhibition of butyrylcholinesterase (BuChE). Using mass spectrometry and NMR studies, three active constituents were isolated and identified: 8-hydroxydihydrochelerythrine (1), 8-hydroxydihydrosanguinarine (2), and berberine (3). Compounds 1-3 showed potent inhibitory activity against AChE, with IC50 (microM) values of 0.61-1.85. Compound 1 exhibited competitive and selective inhibition for AChE. PMID- 17077539 TI - Particulate and microbial contamination in in-use admixed intravenous infusions. AB - We compared particulate and microbial contamination in residual solutions of peripheral intravenous admixtures after the termination of drip infusion between intravenous fluids admixed with glass ampoule drugs and those admixed with pre filled syringe drugs. The mean number of particles>or=1.3 microm in diameter per 1 ml of residual solution was 758.4 for fluids (n=60) admixed with potassium chloride in a glass ampoule (20 ml volume), 158.6 for fluids (n=63) admixed with potassium chloride in a pre-filled syringe (20 ml volume), 736.5 for fluids (n=66) admixed with sodium chloride in a glass ampoule (20 ml volume), 179.2 for fluids (n=15) admixed with sodium chloride in a pre-filled syringe (20 ml volume), 1884.5 in fluids (n=30) admixed with dobutamine hydrochloride in 3 glass ampoules (5 ml volume), and 178.9 (n=10) in diluted dobutamine hydrochloride in pre-filled syringes (50 ml volume: For these samples alone, particulate and microbial contamination were evaluated in sealed products.) Thus, for potassium chloride or sodium chloride for injection, the number of particles>or=1.3 microm in diameter in the residual intravenous solution was significantly higher for fluids admixed with glass ampoule drugs than for those admixed with pre-filled syringe drugs (p<0.0001). For dobutamine hydrochloride for injection, the number of particles>or=1.3 microm in diameter in the residual intravenous solution was estimated to be higher for fluids admixed with its glass ampoule drug than for those admixed with its pre-filled syringe drug. Observation of the residual solutions of fluids admixed with potassium chloride, sodium chloride, or dobutamine hydrochloride in glass ampoules using an electron microscope with an X ray analyzer showed glass fragments in each residual solution. Therefore, for the prevention of glass particle contamination in peripheral intravenous admixtures, the use of pre-filled syringe drugs may a useful method. No microbial contamination was observed in any of the residual solutions of 5 types of admixture. PMID- 17077540 TI - Effects of isopropyl palmitate on the skin permeation of drugs. AB - The model penetrants oxaprozin, nimesulide, gliclazide, and ribavirin, because of their different lipophilicities, were selected to assess the enhancing activity of pre-treatment solutions consisting of isopropyl palmitate (IP) in ethanol (5%, 10%, 15%and 20%, w/w, respectively) across excised rat skin using Franz diffusion cells and HPLC detection. All pre-treatment solutions produced a significant increase in the flux and permeation of all four penetrants (p<0.001) and a relationship between penetrant lipophilicity and enhancement effect was observed. The general order of IP effectiveness at concentration was 20%>15%>10%>5% (w/w). The lag-time of drugs did not significantly change except for ribavirin. PMID- 17077541 TI - Correlation between plasma glucagon-like peptide 2 levels and proliferative makers in small intestinal injury in rats induced by methotrexate administration. AB - Glucagon-like peptide 2 (GLP-2) is a potent intestinal epithelium-specific growth factor that has been shown to reduce the severity of inflammatory disorders of the intestine in rodent models. We examined whether a relationship exists between plasma level of GLP-2 and the degree of intestinal injury induced by chemotherapeutic agents in the rat. Methotrexate (MTX) was administrated orally for 6 consecutive days at doses of 1.25, 2.5, and 5.0 mg/kg body weight per day. Mucosal samples of rat duodenum, jejunum, and ileum were used for assessment of mucosal weight, DNA and protein content. Plasma GLP-2 levels were measured on day 8. MTX significantly reduced body weight. The values of all indices tended to decrease in all segments with increases in MTX dose. Plasma GLP-2 levels were significantly higher in the MTX 2.5 mg/kg/d group (p<0.05) and the MTX 5.0 mg/kg/d group (p<0.01) than in the control group. Correlations were found between plasma GLP-2 levels and mucosal weight, DNA and protein content. We concluded that plasma GLP-2 levels reflect the degree of intestinal injury following MTX administration in this preclinical model. PMID- 17077542 TI - The pharmacokinetics of low-dose thalidomide in Japanese patients with refractory multiple myeloma. AB - Thalidomide has been used for the treatment of refractory multiple myeloma, the dosage in Japan is lower than in other countries; however, there is little information on the pharmacokinetics and their relationship with the drug response. The aim of this study was to characterize the pharmacokinetics of low dose thalidomide in Japanese patients with refractory multiple myeloma, and to examine the relationship between pharmacokinetics and adverse events. On the first and second days, a 100 mg capsule was administered to 8 Japanese patients after breakfast and blood samples were obtained. The plasma concentrations were measured using HPLC and analyzed based on a one-compartment model. If intolerable adverse events were not observed for 14 d, the dose was increased to 200 mg. The average apparent volume of distribution (Vd/F), apparent total clearance (CL/F) and area under the plasma concentration-time curve from 0 to infinity (AUC0 infinity), which were 45.3 l, 5.5 l/h and 21.7 microg.h/ml, respectively, with smaller Vd/F and CL/F and larger AUC0-infinity than in Caucasian populations. This pharmacokinetic difference may explain the dose difference between Japan and other countries. Adverse events were associated with AUC0-infinity, which was best correlated with plasma concentration at 12 h after administration. The 12-h time point was suggested to be a capable indicator for "safety-oriented" therapeutic drug monitoring of thalidomide. PMID- 17077543 TI - Trace determination of steroids causing age-related diseases using LC/MS combined with detection-oriented derivatization. AB - With the rapid shift to an aging society in Japan, age-related diseases, such as osteoporosis, dementia and cancer, are sharply increasing. The measurement of steroids related to these diseases in biological fluids and tissues is useful for elucidation of the nature, diagnosis and treatment of such diseases. LC/MS is considered to be the most promising method for this purpose due to its specificity and versatility, but it sometimes does not demonstrate the required sensitivity for trace amounts of steroids, because steroids have a rather low response using either electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI). To overcome this problem, the author developed detection-oriented derivatization procedures for steroids in LC/MS. For ESI-MS, introduction of a permanently charged moiety is effective. Based on this, 2 hydrazino-1-methylpyridine was developed and used in monitoring prostatic 5alpha dihydrotestosterone, a good index for the follow-up of patients affected by prostate cancer under androgen deprivation therapy and salivary dehydroepiandrosterone, which is now often designated as an anti-aging hormone. A proton-affinitive Cookson-type reagent, 4-[2-(6,7-dimethoxy-4-methyl-3-oxo-3,4 dihydroquinoxalyl)ethyl]-1,2,4-triazoline-3,5-dione, was used for the determination of 1alpha-hydroxyvitamin D3 [1alpha(OH)D3], a synthetic prodrug of the active form of vitamin D3, in human plasma, and this new LC/positive-APCI-MS method enabled the pharmacokinetic study of 1alpha(OH)D3 in humans. Electron capture APCI-MS based on derivatization with 2-nitro-4 trifluoromethylphenylhydrazine was used for the analysis of neurosteroids, which affect brain excitability through action at the neurotransmitter receptors. With this method, the stress-induced rapid biosynthesis of pregnane-type neurosteroids in rat brains was demonstrated. PMID- 17077544 TI - Particle design of three-component system for sustained release using a 4-fluid nozzle spray-drying technique. AB - We prepared composite particles of acetaminophen (Act) with chitosan (Cht) and hydroxypropylmethylcellulose phthalate (HPMCP) as a carrier using a newly developed 4-fluid nozzle spray-dryer. Cht dissolves in acid solutions and forms a gel, but it is insoluble in alkaline solutions. On the other hand, HPMCP is insoluble in acid solutions, but it dissolves in alkaline solutions. Therefore, we tested a preparation of controlled release composite particles using the characteristics of these carriers. Act and Cht mixtures in prescribed ratios were dissolved in an acid solution. Composite particles of Act and HPMCP in prescribed ratios were dissolved in alkaline solutions. We evaluated the composite particles of the three components (Act, Cht, and HPMCP) by preparing solid dispersions using a 4-fluid nozzle spray-dryer. Observation of particle morphology by scanning electron microscopy (SEM) revealed that the particles from the spray drying process had atomized to several microns and had all become spherical. We investigated the physical properties of the composite particles by powder X-ray diffraction, differential scanning calorimetry, and dissolution rate analysis to clarify the effects of crystallinity on the dissolution rate. Powder X-ray diffraction peaks and the heat of fusion of Act in the spray-dried samples decreased in proportion to the carrier content, indicating that the drug was amorphous. These results indicate that the Act-Cht-HPMCP system formed a solid dispersion. Furthermore, we investigated the interaction between the drug and the carrier using FT-IR analysis. FT-IR spectroscopy of the Act solid dispersions suggested that the Act carbonyl and Cht amino groups formed a hydrogen bond. On the other hand, interaction by hydrogen bond was observed between the carbonyl group of HPMCP with the amino group of Act. In the three-component Act-Cht-HPMCP system, the 4-fluid nozzle spray-dried preparation with a mixing ratio of 1 : 2.5 : 2.5 obtained sustained release preparation in all pH test solutions. PMID- 17077545 TI - Comparative study of chemical constituents of rhubarb from different origins. AB - A comparative study of the pharmacologically active constituents of 24 rhubarb samples, which were identified genetically as Rheum tanguticum, 3 intraspecies groups of R. palmatum and R. officinale, was conducted using reversed-phase high performance liquid chromatography (HPLC) methods. Thirty compounds belonging to anthraquinones, anthraquinone glucosides, dianthrones, phenylbutanones, stilbenes, flavan-3-ols, procyanidins, galloylglucoses, acylglucoses, gallic acid, and polymeric procyanidins were analyzed quantitatively. The drug samples derived from the same botanical source showed similar chromatographic profiles, and the comparable specific shape that appeared in the 10-directed radar graphs constructed on the basis of the results of quantitative analysis indicated the relationship between chemical constituent patterns and genetic varieties of rhubarb samples. PMID- 17077546 TI - Optimization and physicochemical characterization of thermosensitive poloxamer gel containing puerarin for ophthalmic use. AB - The purpose of this study was to systematically optimize an ophthalmic thermosensitive poloxamer analogs gel containing puerarin that was a free flowing liquid below the room temperature and could shift to a gel with an eligible gel strength and bioadhesive force in physiological condition (dilution by the simulated tear fluid and at 35.0 degrees C). A two-factor, five-level central composite design (CCD) was employed to the optimization procedure. The effect of formulation variables (the w/v concentration of poloxamer 407 (X1) and poloxamer 188 (X2)) on a number of response variables (the gelation temperature before (Y1) and after (Y2) the simulated tear fluid diluted, the difference between them (Y3)) was systemically investigated. A second order polynomial equation was fitted to the data. The resulting equation and response surface plots were used to predict the responses in the optimal region. Finally, 21.0% (w/v) poloxamer 407 and 5.0% (w/v) poloxamer188 were chosen as the optimal poloxamer gel matrix. The influence of the other ingredients on the physicochemical properties of the formulation was also investigated. Hydroxypropyl-beta-cyclodextrin (HPCD) enhanced the gelation temperature and reduced the gel strength and the bioadhesive force, while puerarin and benzalkonium chloride (BC) had a comparatively smaller influence. All the isotonicity agents studied had the gelation temperatures lowered, and the gel strengths and the bioadhesive forces enhanced. But only sodium chloride appears to be a promising isotonicity agent for the poloxamer gel containing puerarin, HPCD and BC. PMID- 17077547 TI - Novel approach to DPI carrier lactose with mechanofusion process with additives and evaluation by IGC. AB - The effect of lactose carrier surface property on the inhalation profile of dry powder inhaler (DPI) was evaluated using a micronized drug (Compound A) by inverse gas chromatography (IGC). Mechanofusion with magnesium stearate (Mg-St) or sucrose stearate increased the fine particle fraction (FPF), considered to be due to decrease in the interaction between Compound A and the lactose carrier. The effect of Compound A concentration on FPF was smaller in mechanofusion processed lactose compared to intact lactose, especially when processed with Mg St. The relationship between the IGC parameters of the lactose and FPF was also investigated. FPF increased as both the dispersive component of the surface energy and acidity similarity between the lactose carriers and Compound A increased. Although further investigation is necessary, it could be suggested that acidity similarity decreases the interaction between Compound A and lactose, thus contributing to the increase in the FPF. In conclusion, (1) mechanofusion with Mg-St or sucrose stearate could be an effective method to improve FPF of a DPI drug formulation; (2) IGC would be a valuable method to investigate the interaction between a drug and the DPI carrier; and (3) a relationship between surface acidity and inhalation profile was suggested. PMID- 17077548 TI - 4-(Pyrrolidinyl)methoxybenzoic acid derivatives as a potent, orally active VLA-4 antagonist. AB - A novel series of benzoic acid derivatives as VLA-4 antagonists were synthesized. Optimization, focusing on activity and lipophilicity needed for cell permeability, resulted in the identification of 15b and 15e with good activity (IC50 = 1.6 nM each) and moderate lipophilicity (Log D = 2.0, 1.8). Furthermore, 15e demonstrated efficacy in murine asthma model by an oral dose of 30 mg/kg. PMID- 17077549 TI - Structures of new flavonoids, erycibenins D, E, and F, and NO production inhibitors from Erycibe expansa originating in Thailand. AB - A new flavanol, erycibenin D, and two new flavans, erycibenins E and F, were isolated from the stems of Erycibe expansa originating in Thailand. The structures of new flavonoids were elucidated on the basis of chemical and physicochemical evidence. In addition, the inhibitory activities of the isolated constituents from E. expansa on lipopolysaccharide-activated nitric oxide production in mouse peritoneal macrophages were examined. Among the principal constituents, two isoflavones, clycosin (IC50 = 13 microM) and erythrinin B (18 microM), and two rotenoids, deguelin (26 microM) and rotenone (27 microM), were found to show potent inhibitory activity. PMID- 17077550 TI - Synthesis and evaluation of 1-arylsulfonyl-3-piperazinone derivatives as factor Xa inhibitors V. A series of new derivatives containing a spiro[imidazo[1,2 a]pyrazine-2(3H),4'-piperidin]-5(1H)-one scaffold. AB - We have already reported unique compounds containing a N,O-spiro acetal structure as an orally active factor Xa (FXa) inhibitor. This time, we described a N,N spiro acetal structure as an analogue of the N,O-spiro acetal structure for an orally active FXa inhibitor. The synthesis of these analogues could be achieved in a similar fashion to the N,O-spiro acetal synthesis. Consequently, FXa inhibitory activity was increased and more active compounds could be found (M58163: IC50 = 0.61 nM, M58169: IC50 = 0.58 nM). Additionally, the absolute configuration could be determined by X-ray crystallography analysis (M58169: (R) config.). PMID- 17077551 TI - Structures of new cucurbitane-type triterpenes and glycosides, karavilagenins and karavilosides, from the dried fruit of Momordica charantia L. in Sri Lanka. AB - Three new cucurbitane-type triterpene called karavilagenins A, B, and C and five new cucurbitane-type triterpene glycosides called karavilosides I, II, III, IV, and V were isolated from the dried fruit of Sri Lanka Momordica charantia L. (Cucurbitaceae) together with two known cucurbitane-type triterpenes, 19(R) methoxy-5beta,19-epoxycucurbita-6,23-dien-3beta,25-diol and 5,19-epoxycucurbita 6,23-diene-3,25-diol, and nine known cucurbitane-type triterpene glycosides, goyaglycosides-b, -c, and -d, and momordicosides F1, F2, G, I, K, and L. The structures of karavilagenins and karavilosides were elucidated on the basis of chemical and physicochemical evidence. PMID- 17077552 TI - Glycosides of 14,15-seco and 13,14:14,15-disecopregnanes from the roots of Cynanchum sublanceolatum. AB - Thirty-two new 13,14-seco and 13,14:14,15-disecopregnane glycosides were obtained from the roots of Cynanchum sublanceolatum (Asclepiadaceae). These glycosides were confirmed to be possessing five known compounds, cynajapogenin A, glaucogenin-A, 13-hydroxycynajapogenin A, 2alpha-hydroxyanhydrohirundigenin, atratogenin A and one new pregnane, 13-epi-cynajapogenin A, as their aglycones, using both spectroscopic and chemical methods. PMID- 17077553 TI - Solubility prediction of drugs in water-polyethylene glycol 400 mixtures using Jouyban-acree model. AB - A numerical method is proposed for predicting solubility of drugs in water-PEG 400 mixtures based on the Jouyban-Acree cosolvency model. The accuracy of the proposed method is evaluated by computing mean percentage deviation (MPD) and compared with that of log-linear model of Yalkowsky. The overall MPDs of the Jouyban-Acree model and the most accurate version of Yalkowsky's model are 39.8 (+/-46.7) % and 175.8 (+/-266.4) %, respectively, and the mean difference is statistically significant (p < 0.0005). The proposed method produces acceptable residual distribution and the probability of solubility prediction with residual log of solubility <0.5 unit is 0.86. The applicability of the proposed method could be extended for predicting the solubility of drugs in water-PEG 400 mixtures at various temperatures. The impact of various log P values computed using different software is also studied and the results of ANOVA revealed that there are no significant differences between the accuracy of the predicted solubilities employing various log P values. PMID- 17077554 TI - Synthesis and anti-tumor activity of a fluorinated analog of medroxyprogesterone acetate (MPA), 9alpha-fluoromedroxyprogesterone acetate (FMPA). AB - We synthesized 9alpha-fluoromedroxyprogesterone acetate (FMPA) in order to test whether it is a more potent anti-angiogenic agent than medroxyprogesterone acetate (MPA), which has been widely used as a therapeutic agent for breast and endometrium cancers. FMPA was previously synthesized in 10 steps (total yield: 1%). An efficient synthesis of FMPA has been achieved in 6 steps (total yield: 12%). We examined the anti-tumor effect of FMPA, complexed with dimethyl-beta cyclodextrin (DM-beta-CyD), on rat mammary carcinomas induced by 7,12 dimethylbenz[a]anthracene (DMBA). FMPA showed great anti-tumor effect on DMBA induced rat mammary carcinomas. PMID- 17077555 TI - Oxidative photo-decarboxylation in the presence of mesoporous silicas. AB - FSM-16, a mesoporous silica, was found to catalyze oxidative photo decarboxylation of alpha-hydroxy carboxylic acid, phenyl acetic acid derivatives and N-acyl-protected alpha-amino acids to afford the corresponding carbonyl compounds. Furthermore, FSM-16 proved to be re-usable by re-calcination at 450 degrees C after the reaction. PMID- 17077556 TI - Function of an N-heterocyclic carbene ligand based on concept of chiral mimetic. AB - Function of a new N-heterocyclic carbene ligand based on the concept of a chiral mimetic is described. With (4R,5R)-4,5-diphenyl-1,3-dialkyl-4,5-dihydro-3H imidazol-1-ium tetrafluoroborates as N-heterocyclic carbene precursors, Rh catalyzed enantioselective arylation (up to 27% ee) of aromatic aldehydes with arylboronic acids and Pd-catalyzed enantioselective intramolecular alpha arylation (up to 66% ee) of N-(2-bromoaryl)-N-alkyl-2-arylpropanamides are investigated. PMID- 17077557 TI - Xylogranatins A-D, new mexicanolides from the fruit of a Chinese mangrove Xylocarpus granatum. AB - Four new mexicanolides with a Delta(14,15) double bond, named xylogranatins A and D (1-4), were isolated from the fruit of a Chinese mangrove Xylocarpus granatum, together with two known 8,9,30-phragmalin ortho esters, xyloccensins P and Q. Their structures were elucidated on the basis of spectroscopic data, especially 2D NMR techniques including HSQC, HMBC and NOESY. PMID- 17077558 TI - Quantitative structure-activity relationships of selective antagonists of glucagon receptor using QuaSAR descriptors. AB - In the present paper, quantitative structure activity relationship (QSAR) approach was applied to understand the affinity and selectivity of a novel series of triaryl imidazole derivatives towards glucagon receptor. Statistically significant and highly predictive QSARs were derived for glucagon receptor inhibition by triaryl imidazoles using QuaSAR descriptors of molecular operating environment (MOE) employing computer-assisted multiple regression procedure. The generated QSAR models revealed that factors related to hydrophobicity, molecular shape and geometry predominantly influences glucagon receptor binding affinity of the triaryl imidazoles indicating the relevance of shape specific steric interactions between the molecule and the receptor. Further, QSAR models formulated for selective inhibition of glucagon receptor over p38 mitogen activated protein (MAP) kinase of the compounds in the series highlights that the same structural features, which influence the glucagon receptor affinity, also contribute to their selective inhibition. PMID- 17077559 TI - Hepatitis B virus inhibiting constituents from Herpetospermum caudigerum. AB - From the ethanol extract of the seeds of Herpetospermum caudigerum wall, one new lignan compound 1, was isolated and characterized along with three known compounds 2, 3 and 4. The structure elucidation of the isolated new compound was performed on the basis of spectroscopic and chemical evidence. The structures of known compounds were determined by comparison of spectral data and physical data with those previously reported. The activity inhibiting hepatitis b virus was evaluated. Preliminary studies showed that compound 1 and 2 displayed promising inhibitory potential against hepatitis b virus. PMID- 17077560 TI - Sesquiterpene coumarins from the roots of Ferula sinkiangensis and Ferula teterrima. AB - Three new natural sesquiterpene coumarins, isofeterin (1), lehmannolol (3), sinkianone (4), and one known compound, lehmannolone (2), were isolated from the roots of Ferula teterrima and Ferula sinkiangensis. Their chemical structures were established on the basis of spectroscopic analysis, including X-ray crystallography and CD spectrum measurements for determining the absolute configuration of compound 2. PMID- 17077561 TI - New sesquiterpene lactones from the aerial parts of Pseudoelephantopus spicatus. AB - Two new sesquiterpene lactones, spicatolide C (1) and spicatocadinanolide A (2), have been isolated along with the known piptocarphol isomers (3, 4) and one eudismane type sesquiterpene (5) from the EtOAc extract of the aerial parts of Pseudoelephantopus spicatus. The structures and the relative stereochemistries of the new metabolites were determined by spectroscopic methods. PMID- 17077562 TI - Semisynthesis of isetexane diterpenoid analogues and their cytotoxic activity. AB - Isetexane diterpene analogues were semisynthesized from demethylsalvicanol isolated from Perovskia abrotanoides (Labiatae). The structure and cytotoxic activity relationships (SAR) of the natural parent diterpene, demethylsalvicanol, and its semisynthetic analogues were studied by using P388 murine leukemia cells. PMID- 17077563 TI - Cucurbitacin B 2-sulfate and cucurbitacin glucosides from the root bark of Helicteres angustifolia. AB - A new sulfated cucurbitacin, cucurbitacin B 2-sulfate (1) and a new cucurbitacin glucoside, cucurbitacin G 2-O-beta-D-glucopyranoside (2) together with two known cucurbitacin glucosides, arvenin I and arvenin III were isolated from the root bark of Helicteres angustifolia. The structures of these compounds were established on the basis of spectroscopic and chemical evidence. These four compounds taste of strong bitterness. Compound 1 is a first sulfated cucurbitacin found in plants. PMID- 17077564 TI - A facile and clean direct cyanation of heteroaromatic compounds using a recyclable hypervalent iodine(III) reagent. AB - The facile and clean direct cyanating reaction of pyrroles and thiophenes has been achieved using a recyclable hypervalent iodine(III) reagent 1b by a simple solid-liquid separation of the products and the reagent. PMID- 17077565 TI - Scandium-catalyzed carbon-carbon bond formations using alpha-organosulfanyl and organoselanyl-alpha-fluoroacetic acid derivatives. AB - The scandium-catalyzed reactions of alpha-organosulfanyl and organoselanyl-alpha fluoroacetates 1-2, acetamides 3-4 and acetonitrile 5 with soft nucleophiles proceeded to give the products 6a-b, 7a-c, 8a-c, 9a-e in good to high yields. We also successfully performed the scandium-catalyzed intramolecular cyclization reactions and obtained the unique 5-methylene-2-oxotetrahydropyrans 16-17. PMID- 17077566 TI - Efficient synthesis of isothiocyanates based on the tandem Staudinger/aza-Wittig reactions and mechanistic consideration of the tandem reactions. AB - The tandem and stepwise Staudinger/aza-Wittig reactions of several azides were examined in detail. The tandem reaction method (Method I) exhibited superior results in the yield of the corresponding isothiocyanates bearing an electron withdrawing group than the conventional stepwise method (Method II) which involves the sequential treatment of the azides with triphenylphosphine and then carbondisulfide. The mechanistic consideration for both reaction methods was proposed on the basis of the 1H-NMR analyses. PMID- 17077567 TI - Aerobic oxidation of benzyl- and allylic alcohols under visible light irradiation of a fluorescent lamp in the presence of catalytic iodine. AB - Benzyl alcohols and allylic alcohols were found to be oxidized to the corresponding aldehydes in the presence of a catalytic amount of iodine under irradiation of a fluorescent lamp. PMID- 17077568 TI - Dirhodium(II) tetrakis(perfluorobutyrate)-catalyzed 1,4-hydrosilylation of alpha,beta-unsaturated carbonyl compounds. AB - The use of dirhodium(II) catalysts in the 1,4-hydrosilylation of alpha,beta unsaturated ketones and aldehydes was explored. Dirhodium(II) tetrakis(perfluorobutyrate), Rh2(pfb)4, proved to be the catalyst of choice for this process, providing the corresponding silyl enol ethers in high yields. PMID- 17077569 TI - Adrenal insufficiency complicated with antiphospholipid syndrome. PMID- 17077570 TI - Cocoa supplementation for copper deficiency associated with tube feeding nutrition. AB - OBJECTIVE: Because of an increasing number of case reports of copper deficiency associated with long-term tube feeding nutrition in Japan, we identified patients with copper deficiency associated with long-term tube feeding and described the prevalence, clinical data and cocoa treatment for these patients. MATERIALS AND METHODS: We conducted a retrospective study to identify patients who were referred from long-term care institutions and had copper deficiency associated with tube feeding. We reviewed all serum copper concentration data during a 6 year period. We also compared admission and post-treatment peripheral blood counts. RESULTS: Among 210 consecutive admissions with nutritional tube feeding from long-term care institutions (N=210), we identified 13 (6.2%) patients with copper deficiency (aged 46-91; 12 women). All patients had anemia, while most had neutropenia. The range of serum copper concentrations of these patients was 0.1 2.4 microg/L (normal; 6.8-12.8 microg/L). Their feeding formulas revealed a low copper content (5 to 12 microg per 100 kcal of each formula). Cocoa powder was used as the treatment. With cocoa supplements, the blood leukocyte count and hemoglobin significantly improved in all patients. Median leukocyte counts were 1,800 /mm(3)at admission and 6,300/mm(3) at follow-up (p=0.001). Median hemoglobin were 7.0 g/dl at admission and 10.3 g/dl at follow-up (p=0.001). Two patients developed transient tachycardia as a possible adverse effect of cocoa. CONCLUSION: We identified many cases with copper deficiency associated with tube feeding in Okinawa, Japan. Cocoa supplement appeared to be a safe and effective treatment. Increasing the copper content of Japanese tube feeding formulas should be considered for its prevention. PMID- 17077571 TI - Diffuse hepatoid adenocarcinoma in the peritoneal cavity. AB - Hepatoid adenocarcinoma (HAC) is a rare tumor that occurs mostly in the gastrointestinal tract. We encountered a 68-year-old man suffering from HAC with severe ascites. The serum and ascitic alpha-fetoprotein (AFP) levels were markedly high. Computed tomography (CT) revealed that multiple nodular lesions were disseminated only in the peritoneal cavity. Based on laparoscopic and histological findings, this tumor was diagnosed as diffuse HAC which was developed primarily in the peritoneum. Despite treatments with appropriate chemotherapy, he died six months after diagnosis. According to literature updates, this is the first documented case of diffuse HAC which developed in the peritoneum. PMID- 17077572 TI - Successive, rapid recurrences of in-stent restenoses in a woman with angina pectoris. AB - A 62-year-old woman was admitted because of chest pain. Coronary angiography demonstrated stenosis of the left anterior descending coronary artery, where two bare metal stents had been implanted. Over 5 months in-stent restenosis occurred four times, and was treated at first with angioplasty but finally with two sirolimus-eluting stent insertions. Six months after the second insertion, angiography showed restenosis within one stent. The mechanism underlying this succession of rapid restenoses is unknown, but sirolimus-eluting stents appeared to show relative resistance. PMID- 17077573 TI - A pregnant woman with complications of lymphangioleiomyomatosis and idiopathic thrombocytopenic purpura. AB - A pregnant 26-year-old woman developed hemosputum, dyspnea and pneumothorax. Lymphangioleiomyomatosis was suspected based on multiple cystic lesions on chest computed tomography. Additionally, moderate thrombocytopenia occurred during the last trimester. Hyperplasia of megakaryocytes in a bone marrow specimen and a high serum titer of platelet-associated IgG led to a diagnosis of idiopathic thrombocytopenic purpura. High-dose intravenous gammaglobulin promptly restored her platelet count, and the patient successfully gave birth to a healthy baby by cesarean section. After delivery, lymphangioleiomyomatosis was diagnosed by lung biopsy that was obtained during a video-assisted thoracoscopic abscission for recurrent pneumothorax. Underlying lymphangioleiomyomatosis and idiopathic thrombocytopenic purpura may be obviated by pregnancy. PMID- 17077574 TI - Renal segmental hypoplasia, Ask-Upmark kidney, in a patient with adult-onset hypertension. PMID- 17077575 TI - Thymidine phosphorylase gene mutations cause mitochondrial neurogastrointestinal encephalomyopathy (MNGIE). PMID- 17077576 TI - Cutaneous and nodal cryptococcosis in an ATL Patient. PMID- 17077577 TI - The production of a diabetic mouse using constructs encoding porcine insulin promoter-driven mutant human hepatocyte nuclear factor-1alpha. AB - A diabetic mouse model was produced using a mutant human hepatocyte nuclear factor-1alpha gene (HNF1alphaP291fsinsC) regulated by the porcine insulin promoter. The functionality of two different constructs containing HNF1alphaP291fsinsC, termed PD1 and PD2 (cytomegalovirus enhancer minus and plus), were examined in transgenic mice. The blood glucose levels and body weights of the PD1 transgenic mice did not differ from their non-transgenic littermates over the period from 3 to 8 weeks of age. Conversely, the PD2 transgenic mice exhibited hyperglycemia and decreased body weight. Western blot analysis demonstrated that mutant HNF-1alpha protein (HNF1alphaP291), derived from the PD2 transgene, was expressed in the PD2 mice. Morphometric studies of the pancreas of a PD2 mouse revealed that the number of pancreatic islets present was less than that in the non-transgenic mice, indicating disturbed islet neogenesis. These results suggest that impaired insulin secretion in disrupted islets causes hyperglycemia. In addition, the phenotype of PD2 transgenic mice similar to that of the HNF-1alpha gene-deficient mouse, which displays growth retardation and impaired viability. These results indicate that HNF1alphaP291 expression driven by the porcine insulin promoter, together with the cytomegalovirus enhancer, induces a diabetic phenotype in transgenic mice. PMID- 17077578 TI - Calbindin-D9k mRNA expression in the rat uterus following exposure to methoxychlor: a comparison of oral and subcutaneous exposure. AB - Calbindin-D(9k) (CaBP-9k) is a cytosolic calcium-binding protein that is induced by estrogenic compounds possibly through estrogen receptors. We compared CaBP-9k mRNA expression in the uterus with uterotrophic response in immature rats exposed to methoxychlor (MC), an environmental chemical with estrogenic activity. MC was orally or subcutaneously administered to 3-week-old female Sprague-Dawley rats for 3 days. The weights of the uterus and vagina significantly increased in the oral treatment group at a dose of 50, 100 and 200 mg/kg, but those of the subcutaneous (SC) treatment group only increased at 200 mg/kg. Northern blot analysis showed that CaBP-9k mRNA expression was significantly induced in a dose dependent manner at doses of 50, 100 and 200 mg/kg/day in the oral treatment group. SC administration of MC induced significant expression at only a dose of 200 mg/kg/day; this was similar to the uterotrophic response. MC has an estrogenic effect on the uterus as shown by the increase in weight and induction of CaBP-9k mRNA expression, which were much greater following exposure via oral gavage than via the SC route. The strong correlation between the results of in vivo uterotrophic assay and CaBP-9k mRNA expression suggests that CaBP-9k mRNA expression in the rat uterus may be used as an early gene marker for detection of the estrogenic effects of putative environmental chemicals. PMID- 17077579 TI - Applicability of a progesterone-based timed artificilal insemination protocol after follicular fluid aspiration using the ovum pick-up technique in suckled beef cows. AB - We conducted a progesterone-based timed AI protocol after follicular fluid aspiration using the ovum pick-up (OPU) technique to examine its applicability to the suckled beef cow. A total of 19 beef cows were randomly allocated to one of the following three groups based on the number of days postpartum: 13 to 60 days (Group A: suckled; early postpartum period, n=9), 61 to 150 days (Group B: suckled; mid postpartum period, n=6), or 151 to 281 days (Group C: non-suckled; prolonged open period, n=4) postpartum. These cows were treated with follicular fluid aspiration and insertion of a progesterone-releasing intravaginal device (PRID) on day 0. The PRID was removed and 500 microg of cloprostenol was intramuscularly administered on day 7. A dose (100 microg) of fertirelin acetate was injected intramuscularly 48 hours later, and this was followed by a timed AI (TAI) after another 18 hours (day 10). Serum samples were taken on days 0, 7, 9, 10, 12, 17, 24 and 31 for determination of the estradiol-17beta (E(2)) and progesterone concentrations. Pregnancy diagnosis was made by rectal palpation approximately 60 days after TAI. There was no significant difference in the peripheral E(2) concentrations among the three groups during the period of the hormonal treatment. The average progesterone concentrations in Group A on day 17 were significantly higher than those in Group B and exceeded 1.0 ng/ml on day 17 and thereafter. There was no significant difference in the numbers of collected immature oocytes among the three groups. The pregnancy rates in Groups A, B, and C were 77.8% (7/9), 83.3% (5/6) and 50.0% (2/4), respectively. In conclusion, this timed AI protocol is applicable to suckled beef cows within the period of 60 days postpartum. PMID- 17077580 TI - Evidence for existence of cAMP-Epac signaling in the heads of mouse epididymal spermatozoa. AB - Cyclic adenosine 3',5'-monophosphate (cAMP) signaling regulates the expression of fertilizing ability in mammalian spermatozoa. Many articles indicate that this signaling is mediated mainly via protein kinase A. Recently, a guanine nucleotide exchange factor for small G protein Rap1 (an exchange protein directly activated by cAMP: Epac) was discovered as a new mediator of cAMP signaling in somatic cells. The aim of this study was to reveal the existence of cAMP-Epac signaling in mouse spermatozoa. Northern blot analysis and in situ hybridization suggested that Epac1 and Epac2 mRNAs were transcribed in the seminiferous epithelia of the testis. This shows that expression of Epac mRNAs is present in mouse testicular germ cells. Indirect immunofluorescence with specific polyclonal antibodies suggested possible co-localization of Epac1 and Rap1 proteins in the heads of epididymal spermatozoa. Moreover, treatment of epididymal spermatozoa with an Epac-specific cAMP analog, 8-pMeOPT-2'-O-Me-cAMP, induced activation of Rap1, as revealed with a commercial kit for pull-down assay. These results indicate the existence of cAMP-Epac signaling in the heads of mouse epididymal spermatozoa. PMID- 17077581 TI - Successful mouse cloning of an outbred strain by trichostatin A treatment after somatic nuclear transfer. AB - Although the somatic cloning technique has been used for numerous applications and basic research of reprogramming in various species, extremely low success rates have plagued this technique for a decade. Further in mice, the "clonable" strains have been limited to mainly hybrid F1 strains such as B6D2F1. Recently, we established a new efficient cloning technique using trichostatin A (TSA) which leads to a 2-5 fold increase in success rates for mouse cloning of B6D2F1 cumulus cells. To further test the validity of this TSA cloning technique, we tried to clone the adult ICR mouse, an outbred strain, which has never been directly cloned before. Only when TSA was used did we obtain both male and female cloned mice from cumulus and fibroblast cells of adult ICR mice with 4-5% success rates, which is comparable to 5-7% of B6D2F1. Thus, the TSA treatment is the first cloning technique to allow us to successfully clone outbred mice, demonstrating that this technique not only improves the success rates of cloning from hybrid strains, but also enables mouse cloning from normally "unclonable" strains. PMID- 17077582 TI - Selection of donor nuclei in somatic cell-mediated gene transfer using a co transfection method. AB - In this study, we introduced a co-transfection method for the selection of donor nuclei in somatic cell-mediated nuclear transfer. Two vectors were constructed in our experiment. One was pMSCV-GFP carrying the neomycin-resistant gene (Neo(r)) and the green fluorescent protein (GFP) reporter gene; the other was pBC1-GFP carrying the mammary gland-specific promoter and target gene GFP. Ovine adult fibroblasts were co-transfected with pMSCV-GFP and pBC1-GFP. The data from this work demonstrated that the GFP genes in both vectors could successfully co integrate into the genomes of ovine adult fibroblasts in three of the four transgenic cell clones assayed. Furthermore, PCR analysis of transgenic embryos proved that the GFP genes in both vectors could co-integrate into the genomes of the reconstructed embryos. Subsequently, analysis of the developmental rate of the reconstructed embryos after nuclear transfer indicated that the blastocyst rate from the co-transfected donor cells was similar (approximate 8 percent) to that from individual pMSCV-GFP transfected donor cells. The influence of co transfection resulting in modification of donor nuclei on development of reconstructed embryos was also investigated. The results of flow cytometric analysis indicated that the co-transfected ovine fibroblasts had similar quiescent characteristics in terms of cell cycle (G0+G1 percent: 73.20 +/- 4.04) to the individual pMSCV-GFP transfected fibroblasts (G0+G1 percent: 70.77 +/- 1.19) after they were treated with serum starvation for five days. Our results suggest that the co-transfection method can be used for selection of donor cell clones in somatic cell-mediated gene transfer experiments. It can be potentially extended to applications related to expression of functional protein in mammary glands and other transgenic research relevant to nuclear transfer. PMID- 17077583 TI - Effect of danazol on NK cells and cytokines in the mouse uterus. AB - Danazol, which has been used as a medicine for endometriosis, has a valid effect in pretreatment of patients receiving in vitro fertilization and embryo transfer, although its reproductive mechanism remains unclear. BALB/c mice were subcutaneously injected with danazol for 2 weeks. Blood and uteri were collected and cytokines were assayed. Following danazol treatment, an increase in pregnancy ratio was evident that was accompanied by up-regulation in serum macrophage colony stimulating factor (M-CSF). RT-PCR analysis revealed that expression of M CSF and Ly49, a phenotypic marker of natural killer (NK) cells, was up-regulated in the uteri of the danazol-treated mice. In immunohistochemical analysis, M-CSF and Ly49, together with alpha5 integrin, were clearly detected in the endometrium of the danazol-treated mice with very similar pattern of localization. These results suggest that danazol has an effect to promote pregnancy that induces recruitment of NK cells and a concomitant increase in the expression of M-CSF and alpha5 integrin in the uterus. PMID- 17077584 TI - Possible mechanism for testicular focal necrosis induced by hCG in rats. AB - Possible mechanisms for testicular focal necrosis induced by human chorionic gonadotropin (hCG) were examined in Fischer 344 rats. A single s.c. injection of 2000 IU/kg hCG produced focal necrosis 2 days later in testicular tissues such as the seminiferous tubules in the frontal lower part of the testis (FLPT) of 11 week-old F344/Jcl rats. This hCG-induced necrosis was suppressed by an oral treatment (concomitant or delayed by 3 hr) with cyclooxygenase inhibitors (indomethacin, rofecoxib) or prostaglandin (PG) receptor blocker (AA-2414). Focal necrosis was also induced by intratesticular injection of PGF(2alpha) or PGE(2) with this necrosis suppressed by previous oral treatment with AA-2414, and the PGF(2) level in the testis increased 4 hr after hCG treatment. These findings suggested that de novo synthesis of PGs beginning at 3-4 hr was responsible for induction of necrosis. No necrosis was induced by hCG in the Leydig cell-devoid testis produced by ethane dimethanesulfonate treatment. Necrosis of spontaneously induced Leydig cell tumor mass was also induced by hCG, suggesting that Leydig cells are responsible for induction of necrosis. An injection of dye into the testicular artery and laser Doppler flowmetry revealed a continuous reduction of blood flow at the FLPT at 6-48 hr after hCG treatment; contrary to this, the upper part showed an early recovery from the reduced flow. From these results, the mechanism of the hCG-induced necrosis was concluded to be: 1) hCG stimulates Leydig cells to synthesize PGs de novo; 2) PGs induce the intratesticular arteries to contract in the FLPT; and 3) obstruction of blood flow (ischemia) for more than 12 hr induced focal necrosis in the testis. PMID- 17077585 TI - High amount of dinophysistoxin-3 in Mytilus chilensis collected in Seno de Reloncavi, chile, during massive human intoxication associated with outbreak of Vibrio parahaemolyticus. AB - This study describes the detection of high amount of 7-O-acyl-derivative dinophysistoxin-1 (Dinophysistoxin-3) in filter bivalves collected on February 2005 in the Seno de Reloncavi, Puerto Montt City, Southern Chile, in the same period of time where an intoxication episode was associated with the presence of Vibrio parahaemolyticus in shellfish. The Diarrhetic Shellfish Poisoning (DSP) mouse bioassay of mussel extract samples, performed as described for regulatory testing, were negative to DSP toxins. Therefore, the same mussel samples collected from 8 places of Seno de Reloncavi were then analyzed by the HPLC-FLD method with pre-column derivatization procedure for DSP toxins. The samples showed mainly 7-O-acyl-derivative dinophysistoxin-1 (Dinophysistoxin-3) in concentrations ranging from 190.3 +/- 6.8 to 311.1 +/- 4.8 ng of DSP toxin/g hepatopancreas and less amounts of Dinophysistoxin-1 ranging from 1.9 +/- 1.5 to 11.7 +/- 4.6 ng of DSP toxin/g hepatopancreas. After alkaline hydrolysis of the mussel extracts, 279.4 +/- 7.2 ng of Dinophysistoxin-1 /g hepatopancreas (mean +/ SEM, N=6) were found in mussel extracts (Zone 8). These data showed that these shellfish samples are contaminated with the ester form 7-O-acyl-derivatives of Dinophysisyoxin-1, far beyond the safe regulatory limit. This paper also shows a direct relation between lipid content in the mussel tissue extracts and the levels of Dinophysistoxin-3. The 7-O-acyl-derivative dinophysistoxin-1 ester was the only compound associated with DSP toxins detected in the shellfish samples, and in view of the fact that metabolic transformation of Dinophysistoxin-3 into Dinophysistoxin-1 in humans has recently been described in the literature, the consumption of shellfish contaminated with 7-O-acyl-derivatives dinophysistoxin-1 could be a major reason that explains the diarrhetic symptoms shown by the intoxicated patients. PMID- 17077586 TI - Physicochemical and cell-based approach for early screening of phospholipidosis inducing potential. AB - Some of the principal requisites of toxicity screening methods in drug discovery are their ease to perform and high throughput, as well as the possibility to predict the occurrence of clinical events. Phospholipidosis is one of the toxicities often induced by potential drugs. Several physicochemical methods for the prediction of phospholipidosis have been reported. The purpose of the present study was to examine the predictability of methods based on lipophilicity and charge parameters. We employed a test set of 33 compounds including 11 in-house compounds. The phospholipidosis-inducing potential (PLIP) of the test set compounds was determined by the fluorescence-labeled lipid accumulation assay using isolated rat hepatocytes. This assay was verified by transmission electron microscopy (EM). The usefulness of the ClogP - most basic pK(a) (pK(a) -MB) plot to the PLIP of compounds was examined. This plot was unable to predict the PLIP of zwitterions. In order to improve its predictability, the net charge of a given molecule (NC) was introduced instead of pK(a) - MB, since the NC corresponds directly to the ionization state of compounds in the organelles. Compounds with high ClogP (> 1) and high NC (1< or =NC< or =2) tended to be positive. This finding was also confirmed using 30 additional validation set compounds obtained from the literature. The ClogP - NC plot differentiated positive and negative compounds with more than 98% accuracy (62/63), indicating its usefulness in drug discovery. PMID- 17077587 TI - Contribution of intestinal calcium absorption to 1,25-dihydroxyvitamin D3-induced calcium action in the total parenteral nutrition rat. AB - To investigate the contribution of intestinal calcium (Ca) absorption to 1,25 dihydroxyvitamin D(3) (1,25(OH) (2)D(3))-induced Ca action, we assessed parameters related to Ca metabolism after a single dosing of 1,25(OH)(2)D (3) in the total parenteral nutrition (TPN) solution or 5% D-mannitol (MAN) solution treatment with rats. Animals were divided into 6 groups (vehicle, 100 microg/kg p.o. and 25 microg/kg i.v.; n=8) in Experiment 1 and 8 groups (vehicle, 1, 10 and 100 microg/kg p.o.; n=6) in Experiment 2 at TPN or MAN solution treatment. In both experiments, the parameters related to Ca metabolisms, urinary Ca and urinary deoxypyridinoline on 0-24 hr or serum Ca, osteocalcin and parathyroid hormone at 24 hr were measured after 1,25(OH)(2)D (3) dosing. 1,25(OH)(2)D(3) related increased urinary Ca or serum Ca were observed in both experiments. Decrease rates in change of urinary Ca in TPN solution treatment rats were 36.3% (100 microg/kg p.o.) or 47.1% (25 microg/kg i.v.) of MAN solution treatment rats in Experiment 1, and 29.0% (1 microg/kg), 56.2% (10 microg/kg) or 35.3% (100 microg/kg) of MAN solution treatment rats in Experiment 2. Decrease rates in change of serum Ca at 72 hr in TPN solution treatment rats were 57.3% (100 microg/kg p.o.) or 44.5% (25 microg/kg i.v.) of MAN solution treatment rats in Experiment 1, and were 57.0% (100 microg/kg) of MAN solution treatment rats in Experiment 2. There were no differences in the change of serum Ca in the 1,25(OH)(2)D(3) 1 or 10-microg/kg group in Experiment 2. Our results suggest that differences in urinary Ca or serum Ca between MAN solution treatment rats and TPN solution treatment rats express the contribution of intestinal Ca absorption to 1,25(OH)(2)D(3)-induced Ca action in the conditions of the study. PMID- 17077588 TI - DNA microarray analysis of pulmonary fibrosis three months after exposure to paraquat in rats. AB - Although paraquat (PQ) is known to induce pulmonary fibrosis, how it does so is not entirely clear. To elucidate the mechanisms involved, the profile of gene expression in the lung at three months after exposure to PQ (7 mg/kg, s.c., daily for eight administrations) was investigated in rats using a DNA microarray. Changes in gene expression that were considered to reflect damage to the lung, a change in the balance of electrolytes and fluid, and alveolar remodeling were observed. The products of these genes were: CSF-1 receptor, which is a receptor of inflammatory cytokines that activates monocyte/macrophages; TGF-beta type II receptor, which is a receptor of TGF-betas involved in wound healing and fibrosis; a subunit of Na+/K(+)-ATPase, an amiloride-sensitive cation channel, and a subunit of the potassium channel, all of which regulate the alveolar fluid balance and play a role in clearing lung edema; the adenosine A2a receptor, which has a protective function in the lung and interacts with dopamine D1 and D2 receptors to regulate the function of amiloride-sensitive cation channels; cofilin, which is involved in the depolymerization and cleavage of actin filaments; LIM motif-containing protein kinase 1, which negatively regulates the activity of cofilin; SHPS-1, which regulates the integrin-mediated reorganization of the cytoskeleton; and sodium channel beta 2, which is involved in cell adhesion and migration. These results indicate that PQ-induced pulmonary fibrosis does not merely terminate as cicatrices three months after the discontinuation of PQ treatment, but that dynamic functional change continues in the lung. PMID- 17077589 TI - Lack of hinokitiol (beta-thujaplicin) carcinogenicity in F344/DuCrj rats. AB - Chronic toxicity and carcinogenicity of hinokitiol (beta-thujaplicin), used as an antibiotic and fungicidal agent of a food additive, was examined in both sexes of F344/DuCrj (F344) rats. In this chronic toxicity study, groups of 10 rats of each sex were given a diet containing hinokitiol at doses of 0, 0.005, 0.015 and 0.05% for 52 weeks. No treatment-related adverse effects were noted in the survival rate, general condition, body weights, food consumption, urinalysis, hematology and clinical chemistry. Slight but significant elevation of spleen and liver weights was noted in both sexes given 0.05% hinokitiol, along with an increase in hemosiderin deposits in male spleens, related to chelator binding of iron, together with slight centrilobular hypertrophy of male hepatocytes. However, these alterations were negligible and not toxicologically significant. In the carcinogenicity study, groups of 50 female and 50 male rats were given a diet containing hinokitiol at doses of 0, 0.005, 0.015 and 0.05% (excluding 0.005% in females). No treatment-related changes in survival rate, general condition, body weight, food consumption, hematology and organ weights were noted. Detailed histopathological examination revealed no treatment-related increase in the incidences of any neoplastic lesions. The results demonstrate that hinokitiol is not carcinogenic in F344 rats of either sex. PMID- 17077590 TI - Sirc-cvs cytotoxicity test: an alternative for predicting rodent acute systemic toxicity. AB - An in vitro crystal violet staining method using the rabbit cornea-derived cell line (SIRC-CVS) has been developed as an alternative to predict acute systemic toxicity in rodents. Seventy-nine chemicals, the in vitro cytotoxicity of which was already reported by the Multicenter Evaluation of In vitro Toxicity (MEIC) and ICCVAM/ECVAM, were selected as test compounds. The cells were incubated with the chemicals for 72 hrs and the IC(50) and IC(35) values (microg/mL) were obtained. The results were compared to the in vivo (rat or mouse) "most toxic" oral, intraperitoneal, subcutaneous and intravenous LD(50) values (mg/kg) taken from the RTECS database for each of the chemicals by using Pearson's correlation statistics. The following parameters were calculated: accuracy, sensitivity, specificity, prevalence, positive predictability, and negative predictability. Good linear correlations (Pearson's coefficient; r>0.6) were observed between either the IC(50) or the IC(35) values and all the LD(50) values. Among them, a statistically significant high correlation (r=0.8102, p<0.001) required for acute systemic toxicity prediction was obtained between the IC(50) values and the oral LD(50) values. By using the cut-off concentrations of 2,000 mg/kg (LD(50)) and 4,225 microg/mL (IC(50)), no false negatives were observed, and the accuracy was 84.8%. From this, it is concluded that this method could be used to predict the acute systemic toxicity potential of chemicals in rodents. PMID- 17077591 TI - QT-RR relationships and suitable QT correction formulas for halothane anesthetized dogs. AB - Several QT correction (QTc) formulas have been used for assessing the QT liability of drugs. However, they are known to under- and over-correct the QT interval and tend to be specific to species and experimental conditions. The purpose of this study was to determine a suitable formula for halothane anesthetized dogs highly sensitive to drug-induced QT interval prolongation. Twenty dogs were anesthetized with 1.5% halothane and the relationship between the QT and RR intervals were obtained by changing the heart rate under atrial pacing conditions. The QT interval was corrected for the RR interval by applying 4 published formulas (Bazett, Fridericia, Van de Water, and Matsunaga); Fridericia's formula (QTcF = QT/RR(0.33)) showed the least slope and lowest R(2) value for the linear regression of QTc intervals against RR intervals, indicating that it dissociated changes in heart rate most effectively. An optimized formula (QTcX = QT/RR(0.3879)) is defined by analysis of covariance and represents a correction algorithm superior to Fridericia's formula. For both Fridericia's and the optimized formula, QT-prolonging drugs (d,l-sotalol, astemizole) showed QTc interval prolongation. A non-QT-prolonging drug (d,l-propranolol) failed to prolong the QTc interval. In addition, drug-induced changes in QTcF and QTcX intervals were highly correlated with those of the QT interval paced at a cycle length of 500 msec. These findings suggest that Fridericia's and the optimized formula, although the optimized is a little bit better, are suitable for correcting the QT interval in halothane-anesthetized dogs and help to evaluate the potential QT prolongation of drugs with high accuracy. PMID- 17077592 TI - Characterization of an immortalized hepatic stellate cell line established from metallothionein-null mice. AB - Hepatic stellate (HS) cells were isolated from the livers of metallothionein (MT) null and control mice and used to establish IMS/MT(-) and IMS/N cell lines, respectively, using SV40 virus transformation. Cellular morphology, incorporation of vitamin A and expression of alpha-SMA, desmin and SV40 T-antigen were used to confirm that both cell lines were immortal HS cells. The growth rates of both cell lines were similar and there was little difference between cell line sensitivity to zinc. MT-null IMS/MT(-) cells were more sensitive to cadmium and mercury, although both cell lines accumulated almost equal amounts of cadmium during a 24-hr culture period. As HS cells play an important role in hepatic fibrosis and are activated by heavy metals such as cadmium or reactive oxygen, the MT-null HS cell line derived in this study should be a useful experimental model for examination of the role of MT in HS cell activation. PMID- 17077593 TI - Genotoxicity studies OF p-dimethylaminoazobenzene (DAB). AB - The potential genotoxicity of the rodent liver carcinogen p dimethylaminoazobenzene (DAB) was evaluated in compliance with the guidelines for genotoxicity studies of drugs (Notification No. 1604, Nov. 1, 1999, Ministry of Health and Welfare, Japan) and the OECD guidelines for testing chemicals. DAB was clearly positive in both the bacterial reverse mutation test (Ames test) and in vitro chromosomal aberration test in the presence of metabolic activation, whereas it was weakly positive at toxic doses in the rat bone marrow micronucleus test. It has been reported that DAB was clearly positive in in vivo genotoxicity tests, i.e., a mouse alkaline single cell gel electrophoresis (comet) assay and a young rat liver micronucleus test. These results suggest that the test system using the liver is effective for in vivo genotoxicity assessment of chemicals that show mutagenicity in in vitro genotoxicity tests in the presence of metabolic activation. PMID- 17077594 TI - Management of the pain associated with endometriosis: an update of the painful problems. AB - Endometriosis is a condition characterized by ectopic endometrial tissues located outside of the uterus, most commonly found on the pelvic peritoneum or ovary. Endometriosis, which occurs in 7-10% of women in the general population and 71 87% of women with chronic pelvic pain, is associated with dysmenorrhea, chronic pelvic pain, and infertility. There is considerable debate about the effectiveness of various interventions for endometriosis. This review discusses the benefits and drawbacks of pharmacologic and surgical treatments for the pain associated with endometriosis. Laparoscopic surgery has been demonstrated to relieve the pain associated with endometriosis. Hormonal therapies, such as gonadotropin-releasing hormone (GnRH) analogues or the weak androgen danazol, have also been effective at relieving the pain associated with endometriosis. Oral contraceptives appear to be as effective as GnRH analogues for pain relief. Although both surgical and pharmacologic treatments have been effective for relief of the pain associated with endometriosis, the recurrence rate remains significant. The management of pain associated with endometriosis has thus not been satisfied. Larger unified clinical trials are needed to evaluate the effectiveness of new treatments in managing the pain associated with endometriosis. PMID- 17077595 TI - Expression profiles of sex steroid receptors in desmoid tumors. AB - Desmoid tumors are benign fibrous neoplasms which arise from the fibrous tissue of intra- and extra- abdominal sites, but their clinical management is sometimes difficult because of extensive infiltration into the surrounding tissues. Desmoid tumors commonly occur in women, especially after childbirth. Recently, both clinical and experimental findings indicate the possible roles of sex steroids in the development and progression of desmoid tumors but detailed information is still ambiguous. In this study, we first examined immunoreactivity of sex steroid receptors in desmoid tumors (27 cases) by immunohistochemistry and compared the findings with those in reactive self-limiting lesions associated with fibrosis (8 cases). Estrogen receptor (ER) alpha and ERbeta immunoreactivities were detected in 7.4% (2/27) and 7.4% (2/27) of desmoid tumors, respectively. One desmoid tumor expressed both ERalpha and ERbeta. Progesterone receptor (PR)-A and PR-B were detected in 25.9% (7/27) and 33.3% (9/27), respectively, and androgen receptor (AR) in 52.9% (14/27). In reactive lesions with fibrosis, only AR was detected in 37.5% (3/8). Sex steroid receptor mRNAs was further examined by reverse transcription and polymerase chain reaction (RT-PCR) analysis using fresh frozen tissues, demonstrating the expression of PR (PR-A and/or PR-B) and AR mRNAs in eight desmoid tumors examined and all cases of reactive fibrosis. These results indicate that sex steroid hormones might play an important role in the pathogenesis of desmoid tumors and could lead to the introduction of novel hormone therapeutic approaches in managing patients with recurrent desmoid tumors. PMID- 17077596 TI - Thoracic myelopathy in Japan: epidemiological retrospective study in Miyagi Prefecture during 15 years. AB - Thoracic myelopathy is defined as spinal cord compression in the thoracic region, leading to sensory and motor dysfunctions in the trunk and lower extremities, and can be caused by various degenerative processes of the spine. Thoracic myelopathy is rare, and there are many unsolved problems including its epidemiological and clinical features. We have established a registration system of spinal surgeries, which covered almost all surgeries in Miyagi Prefecture, and enrolled the data of 265 patients with thoracic myelopathy from 1988 to 2002. The annual rate of surgery gradually increased and averaged 0.9 per 100,000 inhabitants, which was less than 1/10 of that for cervical myelopathy. About 20 patients with thoracic myelopathy are operated on in Miyagi Prefecture each year. It frequently develops in middle-aged males. About half of the cases were caused by ossification of the ligamentum flavum, followed by ossification of the posterior longitudinal ligament, intervertebral disc herniation and posterior spur. Patients usually noticed numbness or pain in the legs and the preoperative duration was long, averaging 2 years. Its symptomatic similarities to lumbar disorders might cause difficulty in making a correct diagnosis. Since thoracic myelopathy can markedly restrict the activities of daily life, even general physicians should recognize this entity. PMID- 17077597 TI - Reduced exposure to mercury in patients receiving enteral nutrition. AB - The exposure to mercury (Hg) of various groups of people with different dietary backgrounds has been assessed because of its hazardous effects, but little is known about that in patients receiving enteral nutrition. Therefore, we studied the Hg exposure in 25 patients with severe motor disabilities, who received liquid enteral feedings for more than one year, by determining total mercury (T Hg) in their hair samples with inductively coupled plasma-mass spectrometry. The geometric mean of the T-Hg level in hair from the patients was 88 ng/g hair (+/- 1 geometric standard deviation [GSD], 34 - 228 ng/g), whereas that for the control group on a normal diet was 1,900 ng/g (+/- 1 GSD, 1,022 - 3,531 ng/g). The T-Hg levels in the patients' hair were far lower than those in the controls (p < 0.001). The T-Hg levels in the enteral feedings used were below the detection limit of cold-vapor atomic absorption spectrophotometry (< 10 ng/g). The present study has shown that Hg exposure is low in patients receiving enteral nutrition, indicating that food is a primary source of Hg exposure. PMID- 17077598 TI - Improvement of psychiatric symptoms after electroconvulsive therapy in young adults with intractable first-episode schizophrenia and schizophreniform disorder. AB - Schizophrenia is a serious psychiatric disorder that develops mainly in young adults. Electroconvulsive therapy (ECT) is known to be effective and safe in patients with schizophrenia with acute psychotic exacerbation. Because of the shortage of systematic studies, we conducted a prospective naturalistic study to examine the short-term effects of acute ECT and its safety in young adults with medically intractable first-episode schizophrenia. Subjects were seven consecutive patients, 15-35 years of age, with first-episode schizophrenia or schizophreniform disorder (Diagnostic and Statistical Manual of Mental Disorders, 4th edition; DSM-IV), who had failed to respond to neuroleptics. The seven patients were treated with a first course of ECT, and their clinical symptoms were evaluated on the basis of the Brief Psychiatric Rating Scale (BPRS) (18 items, rated 0-6) and Global Assessment of Functioning (GAF) Scale. The GAF Scale is presented in DSM-IV as a means of assessing global functioning of a psychiatric patient. Scores range from 1-100; the higher GAF score indicates the higher global functioning. Adverse effects resulting from acute ECT were also evaluated. The total BPRS score 1 week after the final session improved significantly compared to the total pre-ECT BPRS score. The GAF score also improved significantly compared to the pre-ECT GAF score. There were no adverse effects during the acute ECT course, except for mild delirium. We conclude that ECT may be an effective and safe treatment option for young adults with intractable first-episode schizophrenia. PMID- 17077599 TI - Establishment and characterization of a clonal human extraskeletal Ewing's sarcoma cell line, EES1. AB - Ewing's sarcoma, a small round cell sarcoma arising in soft tissue as well as the bone, is one of the most malignant tumors in children and young adults. Few established cell lines of extraskeletal Ewing's sarcoma (EES) have been reported, which made it difficult to examine the biological features of EES. Therefore, we have established a new clonal cell line of EES. We report its morphological characters, results of chromosomal and immunohistochemical analysis. A piece of tumor obtained from the 18-year-old female patient with EES was xenografted in a nude mouse. In vitro subcultured cells were then obtained from this xenograft. A clonal cell line was subsequently established by limiting dilution and designated EES1. EES1 cells had a doubling time of 24 hours. In the xenografted tumor, the cells expressed vimentin, CD99 (MIC2), neuron specific enolase (NSE) and cytokeratin. The original tumor cells also expressed vimentin, CD 99, and NSE, but was negative for cytokeratin. The morphological and immunohistochemical features of this cell line established, except for cytokeratin expression, were consistent with those of the primary tumor. Cytogenetic analysis of EES1 revealed chromosomal translocation of t(11; 12)(q24;ql2). The chimeric fusion of the Ewing's sarcoma gene in band 22q12 with the Friend leukemia virus integration-1 gene in band 11q24 was also demonstrated. Fluorescence in situ hybridization further confirmed the presence of translocation involving the Ewing's sarcoma gene in both the primary tumor and EES1 cells. In conclusion, we have established a human EES cell line EES1, which will provide a useful model for studying various aspects of human EES. PMID- 17077600 TI - Description and evaluation of an innovative course on learning and study skills for the first year medical students. AB - The purpose of this study is to share educational structure and evaluation results of an innovative course on effective learning and study skills for the first year medical students. In Turkey, undergraduate medical education takes six years and each year nearly 5,000 high school graduates start medical schools. However, many students experience frustration and failure because of their lack in the learning and studying strategies. At the Ege University Faculty of Medicine, preclinical curriculum consists of the body function systems-based teaching blocks. Year one has three blocks. We implemented an effective learning and study skills course at the first and third blocks of the 2003-2004 curriculum. We evaluated the course by students' feedbacks derived from block questionnaires and students' homework performance analysis. At the first block questionnaire, out of 297, 163 students (54.8%) clearly stated that the course positively influenced their learning process. Structured analysis of the first block's students' homework showed that an average of 206 students (69.3%) can sufficiently describe their learning and studying approach, while 218 (73.4%) can identify the strengths and weaknesses of the curriculum. The average student scores were 74 +/- 10 and 68 +/- 11 out of 100 for first and third blocks, respectively. We interpreted these results as students enjoyed the course and learned the content. In conclusion, a course on effective learning and study skills is likely to assist first year medical students in improving their learning and adaptation to the school. PMID- 17077601 TI - Glutamine administration enhances the healing of lung parenchymal injuries and reduces air leakage in rats. AB - Beneficial effects of glutamine on wound healing are well known. Parenchymal injuries in the lung cause air leakage that resolves with wound healing. We aimed to determine the effect of glutamine on the healing of lung injuries. Wistar albino female rats were randomized in three groups. One group (control, n = 7) received intraperitoneal injection of 0.9% sodium chloride (1.5 ml /day), while other group (GLN, n = 7) received glutamine (1.5 g/kg/day), beginning two days prior to the operation for total four days. After thoracotomy, a lung parenchymal lesion was made with a scalpel in the right upper lobe. Only thoracotomy was performed to sham group (n = 4). Air leakage was observed in the isolated lungs of control group, but not GLN and sham groups, at 5 cm H(2)O of positive airway pressure (p < 0.001). The threshold of positive airway pressure for air leakage was 4.85 +/- 0.37 and 19.42 +/- 4.54 cm H(2)O for control and GLN groups, respectively (p < 0.001). For measurement of collagen content in the healing parenchyma, digital images were processed to calculate the stained area percentage (SAP). SAP for immature collagen, a marker for wound healing, was 0.36 +/- 0.18% and 1.48 +/- 0.83% (p = 0.02) in control and GLN groups, respectively, but no significant difference was noted in SAP for mature collagen. The grade of inflammation was not significantly different between control and GLN groups. We conclude that glutamine enhances lung parenchymal healing by increasing immature collagen secretion. PMID- 17077602 TI - Persistent oxidative stress after myocardial infarction treated by percutaneous coronary intervention. AB - Acute myocardial infarction causing cardiac ischemia is responsible for the majority of cardiac related deaths. Medical interventions that ensure rapid reperfusion, such as percutaneous coronary intervention, are aimed to allow myocardial re-oxygenation. However, this generates reactive oxygen species, resembling ischemia-reperfusion type of injury based on oxidative stress. In the present study we monitored dynamic changes of total serum peroxides, total antioxidant capacity and soluble intercellular adhesion molecule-1 as well as the titer of antibodies against oxidized low-density lipoproteins in the blood during the convalescence period of 32 patients with acute myocardial infarction treated by percutaneous coronary intervention. Samples were taken at admittance and at two hours, four hours, three days and seven days following percutaneous coronary intervention. Total antioxidant capacity dropped to 82% (p < 0.05). The titer of antibodies against oxidized low-density lipoproteins transiently decreased within the first three days, and increased afterwards. The values of serum peroxides and soluble intercellular adhesion molecule-1 increased continuously in respect to the initial levels reaching the maximum at the time of release from hospital. These findings indicate a persistent oxidative stress that might be associated with intravascular inflammation in patients during convalescence and release from hospital. PMID- 17077603 TI - Enhanced growth of hepatic hemangiomatosis in two adults after postmenopausal estrogen replacement therapy. AB - Liver hemangiomatosis is defined as extensive hemangioma in the liver. Although hemangioma is the most common hepatic tumor, diffuse hepatic hemangiomatosis is very rare. Most cases of hepatic hemangiomatosis are seen in infancy, but it is extremely rare in adults. This is the first report, showing the enhanced growth of diffuse hepatic hemangiomatosis after hormone replacement therapy. We report herein two unrelated women, 47 and 42 year-old, from different regions of Turkey, who admitted to hospital because of right abdominal pain with diffuse hepatic hemangiomatosis, developed after hormone replacement therapy for menopause. The patients were healthy, except for hemangiomatosis, and their physical examination, routine laboratory tests, and tumor marker levels were within normal limits. It should be noted that their abdominal ultrasonography was normal before hormone therapy, but ultrasonography on admission revealed numerous, ill defined, diffusely located liver nodules in both patients. Dynamic magnetic resonance imaging and scintigraphy have revealed that these lesions are compatible with hemangiomatosis. These results suggest that hepatic hemangiomatosis was induced by estrogen therapy. Consequently, hormone replacement therapy was discontinued, and the patients were followed up for 3 years. Their physical examination and blood chemistry, including liver enzymes, remained within normal range, and the follow-up examination with ultrasonography showed no changes in size of lesions. Because of the possible association of hemangioma with estrogen administration, decisions should be made carefully about estrogen therapy for patients who already have hemangioma, and the periodic ultrasonograpy examination should be planned to detect possible new growth of liver hemangiomatosis. PMID- 17077604 TI - Contrast-enhanced dynamic magnetic resonance imaging of primary pulmonary leiomyosarcoma. AB - An abnormal opacity was found incidentally on chest radiography in a 19-year-old Japanese man with lumbago. Chest computed tomography (CT) revealed a 55 x 40 mm well-defined mass with smooth margins in the lower lobe of the right lung (S6). Contrast-enhanced dynamic CT showed that the lesion was not enhanced, suggesting a cystic lesion, such as a bronchogenic cyst. However, contrast-enhanced dynamic magnetic resonance imaging (MRI) revealed enhancement of the lesion, which had a clear margin and homogenous consistency, suggesting a benign pulmonary solid tumor. Video-assisted thoracoscopic surgery, S6 segmentectomy, was performed, disclosing the lesion to be solid. Histopathologically, the tumor consists of spindle cells with ample eosinophilic cytoplasm arranged in an interlacing pattern. Immunohistochemical staining revealed positive desmin reactivity but was negative for alpha smooth muscle actin and HHF-35 (muscle specific actin), indicating atypical phenotypic differentiation. The lesion was accordingly diagnosed as leiomyosarcoma with low grade malignancy, and right lower lobectomy was performed. The patient has remained free from recurrence for 18 months without any additional chemotherapy or radiation therapy. Contrast-enhanced dynamic MRI should be carried out to facilitate accurate preoperative diagnosis and selection of the correct surgical plan. PMID- 17077605 TI - Recurrent Crow-Fukase syndrome associated with increased serum levels of vascular endothelial growth factor: a case report and review of the literature. AB - Crow-Fukase syndrome (CFS) is a rare multi-system disorder, characterized by polyneuropathy, organomegaly, endocrinopathy, M-proteins, skin changes and anasarca, with or without myeloma. The pathophysiology, diagnosis, and treatment of CFS are controversial. CFS may be associated with the overproduction of vascular endothelial growth factor (VEGF). However, there have been no reports of monitoring the serum VEGF level after recurrence, to the best of our knowledge. We report a 54-year-old man with CFS presenting with a 3-year history of ascites, anasarca, weakness of the lower extremities, and plasmacytoma in the scapula. At the initial examination, the VEGF level was 1,590 pg/ml (the VEGF level of a healthy control, 78.4 +/- 75.2 pg/ml). After initial treatment with chemotherapy and irradiation of the affected shoulder, the VEGF level decreased to 154 pg/ml and the symptoms disappeared. Twenty one months later, gate disturbance and anasarca recurred, and the VEGF level was over 2,000 pg/dl. After total scaplectomy, the VEGF level decreased to 730 pg/dl and the symptoms disappeared. The serum level of VEGF well correlated to the clinical course of the patient. In conclusion, measurement of the VEGF level is useful for diagnosing CFS and for monitoring its clinical course. PMID- 17077606 TI - [Recent advance in development of viral and non-viral vectors for gene therapy]. PMID- 17077607 TI - [Characterization of adenovirus serotype 35 vectors using genetically modified animals and non-human primates]. AB - Recombinant Adenovirus (Ad) vectors are considered to be a promising gene delivery vehicle of high utility because they are easy to construct, can be produced at high titers, and efficiently transduce various types of cells. Ad vectors commonly used in the world, including clinical trials, are composed of Ad serotype 5 (Ad5), which belongs to subgroup C. In recent years, however, it has become apparent that Ad5 vectors have some drawbacks, such as high seroprevalence of anti-Ad5 antibodies in adults and low transduction efficiencies of Ad5 vectors in cells lacking a primary receptor for Ad5, coxsackievirus and adenovirus receptor (CAR). To overcome these limitations of Ad5 vectors, we have developed a novel type of Ad vector, which is composed of Ad serotype 35 (Ad35), belonging to subgroup B. Ad35 vectors recognize human CD46, not CAR, as a cellular receptor for infection. Human CD46 is expressed in almost all of human cells, leading to a broad tropism of Ad35 vectors to human cells, in contrast, expression of rodent CD46 is limited to the testis. Therefore, in vivo transduction properties of Ad35 vectors are not appropriately evaluated in normal mice. In order to evaluate the in vivo transduction properties of Ad35 vectors, Ad35 vectors were applied to human CD46-transgenic mice and nonhuman primates, which express CD46 in a similar pattern to humans. The data obtained using CD46-transgenic mice and nonhuman primates would provide valuable information towards clinical applications of Ad35 vectors. PMID- 17077608 TI - [Utility of AAV vectors derived from novel serotypes]. AB - AAV vector is derived from nonpathogenic virus and has a number of attractive features as a vector for human gene transfer including safety, broad tissue specificity, and low immunogenicity following gene transfer. Moreover, persistent transgene expression (for years) was demonstrated in multiple animal experiments. For these reasons, applications to a wide spectrum of diseases are expected, and several clinical trials have been conducted. Although it is too early to conclude the outcome, the efficacy of treatment was not sufficiently substantiated in most of the trials despite confirming the safety of the vector. These results are primarily due to low levels of transgene expression. One of the approaches to improve this situation is the use of alternative serotypes of AAV. Traditionally, serotype 2 was considered to be a prototype of AAV, and the majority of studies including human clinical trials have been conducted using this serotype. On the other hand, there are five "classical" serotypes, and several have been additionally discovered from tissues of primates including humans. These serotypes are considered to be valuable resources for vector development to overcome the shortcomings of serotype 2. This review focuses on the difference in expression levels and tissue specificity of various serotype-derived vectors and summarizes current status in the treatment of candidate diseases. PMID- 17077609 TI - [Optimization of in-vivo gene transfer through regulating biological response to vectors]. AB - The spatiotemporal distribution of transgenes determines the therapeutic efficacy of in vivo gene transfer. The important parameters of gene transfer are the level, duration, and cell specificity of expression, and the number of transfected cells. Interaction of vectors with blood cells, antigen-presenting cells, serum proteins, and other biological components affects the tissue distribution of vectors and the profile of transgene expression. Although plasmid DNA is less immunogenic than viral vectors, it can induce inflammatory cytokine release, due mainly to the presence of unmethylated CpG dinucleotides (CpG motifs). It was clearly demonstrated that intravenous injection of a plasmid DNA/cationic liposome complex resulted not only in the induction of inflammatory cytokines, but also in the activation of nuclear factor kappaB (NF-kappaB) in the lung. Insertion of additional NF-kappaB-binding sequences into conventional plasmid DNA resulted in a high transgene expression in the lung, suggesting that the biological response to vectors can be used to increase transgene expression. In a marked contrast to this strategy, long-term transgene expression was achieved by reducing the number of the CpG motifs in plasmid DNA. A plasmid encoding murine interferon (IFN)-beta or IFN-gamma with reduced numbers of CpG motifs was highly effective in inhibiting metastatic tumor growth in mice. These results clearly demonstrate the importance of the regulation of biological responses to plasmid vectors to optimize plasmid-based in vivo gene transfer. PMID- 17077610 TI - [Development of plasmid DNA-based gene transfer]. AB - Gene therapy based on ultrasound with microbubbles offers a novel approach for the prevention and treatment of variety of diseases. The major development of gene transfer has importantly contributed to intense investigation of the potential of gene therapy in cancer or cardiovascular medicine. The amazing advances in molecular biology have provided a dramatic improvement of the technology that is necessary to transfer target genes into somatic cells. Gene transfer methods have been surprisingly improved. In fact, some of them (retroviral vectors, adenoviral vectors or liposome based vectors, etc.) have been used in the clinical trials already. But some severe side effects were reported in clinical gene therapy using such viral, so people desire safe and efficient clinical gene therapy. Recently, ultrasound-mediated gene transfer has been reported to augment the transfection efficiency and facilitate local gene expression. Interestingly, gene transfer into the fetal central nervous system was successfully achieved by intrauterine injection with microbubble-enhanced ultrasound. Compared to other viral vectors, there are some theoretical advantages including safety, simplicity of preparation, and local gene transfer. Thus, we focused on the development of gene transfer using naked plasmid DNA with an ultrasound or microbubble-enhanced ultrasound method. PMID- 17077611 TI - [Development of non-viral vector based on the quantitative comparison of intracellular trafficking with viral vector]. AB - For the development of efficient gene vector, intracellular processes such as cellular uptake, endosomal release and nuclear delivery must be overcome. Viruses have also evolved and have developed sophisticated mechanisms for controlling intracellular trafficking for the efficient delivery of their genomes to nuclei in host cells for symbiosis. In the light of these mechanisms, various kinds of artificial devices have been developed to overcome the intracellular barriers. However, in the majority of studies, variation of the transfection activity before and after the modification of devices was evaluated, and intracellular trafficking remained unclear. Therefore, it is understand to recognize which of the intracellular barrier should be intensively improved to enhance the transfection activity. To clarify the rate-limited process in the current non viral vector, we compared the intracellular trafficking between adenovirus and LipofectAMINE PLUS. As a result, we found that difference of the transfection efficiency between adenovirus and LipofectAMINE PLUS was dominantly derived from the differences on transcription activity. Therefore it is essential to consider the regulation of the intranuclear events to improve the transfection activity of artificial vector. PMID- 17077612 TI - [Synthesis of fluorine compounds based on special properties of fluorine compounds]. AB - This review describes new syntheses of organofluorine compounds taking advantage of the special properties of fluorine compounds as synthones. The main reactions presented are as follows: 1) Trifluoromethylation of aryl or alkyl halides. Application of this reaction for the synthesis of fluorine analogues of nucleic acid bases is discussed. 2) Some syntheses of trifluoromethyl compounds using the Diels-Alder or the 1,3-dipolar reaction, trifluoromethylated carbene, and photoreaction. 3) The Friedel-Crafts reaction of 3,3,3-trifluoropropene, where linear alkylation occurs due to electronic effect of the trifluoromethyl group in good contrast with the Friedel-Crafts reaction of propene. 4) The ene reaction of trifluoromethylated carbonyl compounds, which work as good enophiles. Application of this reaction to the synthesis of trifluoro analogues of terpenes is discussed. 5) The ene reaction of trifluoromethylated imines. 6) Reaction of halothane, which has a highly acidic hydrogen and two halogens adjacent to the trifluoromethyl group and shows interesting reactivities with various bases and metals to give products with unexpected structures. 7) Reaction of 2-bromo-2,2 difluoroacetate with Cu, where the cross-coupling reaction, Michael-type reaction, and radical reaction for different types of difluoroacetates are presented. 8) Reaction of 2-bromo-2,2-difluoroacetate in the presence of Rh catalyst. This reaction provided a new methodology for the introduction of fluoroalkyl substituents to the alpha-position of alpha,beta-unsaturated ketones. The Rh catalyst solved some difficulties in the introduction of difluoroacetate to carbonyl compounds (Reformatsky reaction). Application of this reaction to imines provided easy access to beta,beta-difluoro-beta-lactams. PMID- 17077613 TI - [Mechanism of cell proliferation--cell cycle, oncogenes, and senescence]. AB - Cell proliferation is regulated through a transition between the G0 phase and cell cycle. We isolated a mammalian temperature-sensitive mutant cell line defective in the function from the G0 phase to cell cycle. Senescent human somatic cells fail to enter into the cell cycle from the G0 phase with stimulation by any growth factor. Telomere shortening was found to be a cause of cellular senescence, and reexpression of telomerase immortalized human somatic cells. Immortalized human somatic cells showed normal phenotypes and were useful not only for basic research but also for clinical and applied fields. The importance of p53 and p21 activation/induction i now well accepted in the signal transduction process from telomere shortening to growth arrest, but the precise mechanism is largely unknown as yet. We found that the MAP kinase cascade and histone acetylase have an important role in the signaling process to express p21. Tumor tissues and cells were found to have strong telomerase activity, while most normal somatic human tissues showed very weak or no activity. Telomerase activity was shown to be a good marker for early tumor diagnosis because significant telomerase activity was detected in very early tumors or even in some precancerous tissues compared with adjacent normal tissues. Telomere/telomerase is a candidate target for cancer chemotherapeutics, and an agent that abrogated telomere functions was found to kill tumor cells effectively by inducing apoptosis whereas it showed no effect on the viability of normal cells. PMID- 17077614 TI - Regulatory mechanism of food factors in bone metabolism and prevention of osteoporosis. AB - Aging induces a decrease in bone mass, and osteoporosis with its accompanying decrease in bone mass is widely recognized as a major public health problem. Bone loss with increasing age may be due to decreased bone formation and increased bone resorption. Pharmacologic and nutritional factors may prevent bone loss with aging, although chemical compounds in food and plants which act on bone metabolism are poorly understood. We have found that isoflavones (including genistein and daidzein), which are contained in soybeans, have a stimulatory effect on osteoblastic bone formation and an inhibitory effect on osteoclastic bone resorption, thereby increasing bone mass. Menaquinone-7, an analogue of vitamin K(2) which is abundant in fermented soybeans, has been demonstrated to stimulate osteoblastic bone formation and to inhibit osteoclastic bone resorption. Of various carotenoids, beta-cryptoxanthin, which is abundant in Satsuma mandarin (Citrus unchiu MARC), has a stimulatory effect on osteoblastic bone formation and an inhibitory effect on osteoclastic bone resorption. The supplementation of these factors has a preventive effect on bone loss induced by ovariectomy in rats, which are an animal model of osteoporosis, and their intake has been shown to have a stimulatory effect on bone mass in humans. Factors with an anabolic effect on bone metabolism were found in extracts obtained from wasabi leafstalk (Wasabi japonica MATSUM), the marine alga Sargassum horneri, and bee pollen Cistus ladaniferus. Phytocomponent p-hydroxycinnamic acid was also found to have an anabolic effect on bone metabolism. Food chemical factors thus play a role in bone health and may be important in the prevention of bone loss with increasing age. PMID- 17077615 TI - [Synthesis and structure-activity relationships of bioactive compounds using sterols]. AB - Sterols are widely and abundantly distributed in nature. It is convenient to utilize them for the preparation of useful compounds such as pharmaceuticals with steroid and secosteroid skeletons. This paper describes the synthesis and structure-activity relationships of naturally occurring active forms of vitamin D analogues, sterols having neurite outgrowth activity, and liver X receptor agonist. The active form of vitamin D(4) showed similar biological activities but had higher affinity to the vitamin D-binding protein compared with the corresponding vitamins D(2) and D(3). This shows that the active form of vitamin D(4) is a good candidate for an agent to replace the active forms of vitamins D(2) and D(3). In the course of screening for low molecular-weight compounds that exhibit neurite outgrowth activity in the culture broth, we found that the natural product dictyosterol showed strong activity. From screening of the analogues, it was found that the double bond between C22 and C23 in the side chain of the sterol is essential for its activity. Ergost-22-ene-1alpha,3beta diol was found to serve as a stronger liver X receptor agonist than 24(S), 25 epoxycholesterol, which regulates the expression of genes involved in lipid metabolism. Structure-function study showed that the 1alpha-hydroxyl group, the saturated steroid structure, and the double bond between C22 and C23 are needed to function as a liver X receptor agonist. PMID- 17077616 TI - [Investigation for proper use of probiotics in Nagasaki University Hospital of Medicine and Dentistry]. AB - Antimicrobial agents sometimes cause the adverse effects of diarrhea and loose stool. Antibiotic-resistant lactic acid bacteria are used to prevent these adverse effects. The bacteria are not resistant to several antimicrobial agents, although the bacterium preparations are sometimes prescribed the antimicrobial agents concomitantly. Therefore this paper reports that the minimal inhibitory concentration of three new antimicrobial agents against antibiotic-resistant lactic acid bacteria were determined using a microdilution method with cation adjusted Mueller-Hinton broth. Furthermore, we investigated antimicrobial agents that are prescribed concomitantly with antibiotic-resistant lactic acid bacterium preparations or a clostridium butyricum preparation. The bacteria were susceptible to the three new antimicrobial agents. Approximately 50% of the bacterium preparations were prescribed alone, and 30% were prescribed concomitantly with antimicrobial agents that show antimicrobial activity against the bacteria. Consequently, we suggest that pharmacists need to confirm prescriptions and to provide more drug information on antibiotic-resistant lactic acid bacterium preparations. PMID- 17077617 TI - [Analysis of the factors associated with drugs remaining in the Diskhaler following inhalation of fluticasone propionate]. AB - Because it is well known that drug remains in the fluticasone propionate Diskhaler (FP-DH) following a single inhalation, the following patient information is recommended. "Please inhale more than once or twice if any drug remains in the device after inhalation". It is believed the inspiratory flow rate of the individual patient has an influence on the amount of drug that remains in the device. If the dosing performance of FP-DH is dependent on inspiratory effort, establishment of a method of inhalation that makes it independent of inspiratory flow rate is important in clinical practice. In the present study, we investigated the influence of various methods of inhalation of drug remaining in the FP-DH. No significant differences were observed regarding the drug remaining in the device among the inhalation times examined (range, 0.5-2.5 s) or the number of inhalations (range, 1-3 times). On the other hand, the amount of drug remaining in the device did decrease by tapping the device before the second inhalation. The results suggest that the amount of drug remaining in the device can be decreased by tapping the device after the first inhalation if the patient's inspiratory flow rate is low. PMID- 17077618 TI - Antinociceptive, anti-inflammatory and antipyretic effects of Solanum nigrum chloroform extract in animal models. AB - AIM: The present study was carried out to evaluate the antinociceptive, anti inflammatory and antipyretic effects of chloroform extract of Solanum nigrum leaves using various animal models. METHODS: The extract was prepared by soaking (1:20; w/v) the air-dried powdered leaves (20 g) in chloroform for 72 hrs followed by evaporation (40 degrees C) under reduced pressure to dryness (1.26 g) and then dissolved (1:50; w/v) in dimethylsulfoxide (DMSO). The supernatant, considered as the stock solution with dose of 200 mg/kg, was diluted using DMSO to 20 and 100 mg/kg, and all doses were administered (s.c.; 10 ml/kg) in mice/rats 30 min prior to tests. RESULTS: The extract exhibited significant (p<0.05) antinociceptive activity when assessed using the abdominal constriction, hot plate and formalin tests. The extract also produced significant (p<0.05) anti inflammatory and antipyretic activities when assessed using the carrageenan induced paw edema and brewer's yeast-induced pyrexia tests. Overall, the activities occurred in a dose-independent manner. CONCLUSION: The present study demonstrated that the lipid-soluble extract of S. nigrum leaves possessed antinociceptive, anti-inflammatory and anti-pyretic properties and confirmed the traditional claims. PMID- 17077619 TI - [Usefulness of small group discussion using KJ method regarding early exposure for first-year pharmaceutical science students]. AB - We evaluated the usefulness of small group discussion using the KJ method (KJ SGD) in regards to early exposure for first-year pharmaceutical science students. Fourteen students were divided into 3 groups. They were asked to discuss the societal role of pharmaceutical science faculty members and then write the results of their discussion. Thereafter, each of the groups presented the contents of their written results to all of the students. Discussion sessions were held on two separate occasions, the first prior to early exposure and the second after early exposure. After receiving early exposure, the students were asked to revise their previous written results. Further, a questionnaire was given to the students following each discussion session, which revealed that more than 92% understood the KJ-SGD method and most answered that it was useful. In addition, all answered that their perceptions were changed after early exposure. Our findings showed that KJ-SGD was useful and that the students were able to effectively understand the contents following early exposure. PMID- 17077620 TI - Determination of liquiritin, naringin, hesperidin, thymol, imperatorin, honokiol, isoimperatorin, and magnolol in the traditional Chinese medicinal preparation Huoxiang-zhengqi liquid using high-performance liquid chromatography. AB - High-performance liquid chromatography was employed to determine the contents of the eight marker components liquiritin, naringin, hesperidin, thymol, imperatorin, honokiol, isoimperatorin, and magnolol in the traditional Chinese medicinal preparation Huoxiang-zhengqi liquid. The separation was performed on a C(18) column by stepwise gradient elution with water-methanol-acetonitrile (0.01 min, 68:30:2; 20 min, 60:38:2; 50 min, 34:64:2; 65 min, 34:64:2; 75 min, 28:70:2; 85 min, 68:30:2) as the mobile phase at a flow rate of 1 ml/min, with UV detection at 283 nm. Eight regression equations showed good linear relationships between the peak area ratio of each marker to internal standard and amounts. The recoveries of the markers listed above were 97.4, 98.5, 97.4, 98.6, 97.8, 99.2, 97.0, and 97.5%, respectively. The repeatability and reproducibility (relative standard deviation) of the method were less than 2.2 and 3.0%, respectively. PMID- 17077621 TI - [Possibility of interactions between prescription drugs and OTC drugs (2nd report)--interaction between levodopa preparation and OTC Kampo medicines for upset stomach]. AB - Our series of studies aimed to examine the possibility of interactions between prescription drugs and over-the-counter (OTC) drugs by monitoring plasma drug concentrations in rats. When a levodopa preparation indicated for patients with Parkinson's disease was administered in combination with Takeda Kampo Ichoyaku K matsu (A), Taisho Kampo Ichoyaku (B), or Kanebo Kampo Ichoyaku H(C), which are OTC kampo medicines for upset stomach, the plasma levodopa concentration-time curves were shifted downward and the AUC for levodopa was significantly lowered. These results indicate that there may be some interactions between the levodopa preparation and these OTC kampo medicines when ingested together, which leads to a reduction in the bioavailability of levodopa. On the other hand, concomitant administration of the levodopa preparation with Takeda Kampo Ichoyaku A-matsu (D) did not alter any of the pharmacokinetic parameters for levodopa. According to the package inserts for the OTC kampo medicines, A, B and C, but not D, contain metallic additives, such as aluminum silicate and magnesium stearate. In addition, combination with a kampo basis of D (Koshaheiisan-ka-shakuyaku) showed no detectable change in levodopa bioavailability. From these results, it was concluded that metallic additives may play an essential role in generating the drug-interaction between levodopa preparation and OTC kampo medicine for upset stomach. PMID- 17077622 TI - Antinociceptive and anti-inflammatory activities of Dicranopteris linearis leaves chloroform extract in experimental animals. AB - The present study was carried out to establish the antinociceptive and anti inflammatory properties of Dicranopteris linearis leaves chloroform extract in experimental animals. The antinociceptive activity was measured using the abdominal constriction, formalin and hot plate tests, while the anti-inflammatory activity was measured using the carrageenan-induced paw edema. The extract, obtained after 72 h soaking of the air-dried leaves in chloroform followed by evaporation under vacuo (40 degrees C) to dryness, was dissolved in dimethyl sulfoxide to the doses of 20, 100 and 200 mg/kg and administered subcutaneously 30 min prior to subjection to the above mentioned assays. The extract, at all doses used, was found to exhibit significant (p<0.05) antinociceptive activity in a dose-dependent manner. However, the significant (p<0.05) anti-inflammatory activity observed occur in a dose-independent manner. As a conclusion, the chloroform extract of D. linearis possesses antinociceptive and anti-inflammatory activity and thus justify its traditional uses by the Malays to treat various ailments. PMID- 17077624 TI - The impact of new comorbidities on nutritional status in continuous ambulatory peritoneal dialysis patients. AB - OBJECTIVE: To study the prevalence and risk factors for malnutrition in a peritoneal dialysis (PD) center with an active PD program. METHODS: We assessed the nutritional status in 205 continuous ambulatory peritoneal dialysis (CAPD) patients, including stable and unstable patients, by subjective global assessment (SGA), dietary diaries and biochemistry index. Serum C-reactive protein (CRP) levels were examined as inflammatory marker. Fluid status including extracellular water (ECW), intracellular water, and total body water (TBW) was evaluated by multiple-frequency bioelectrical impedance analysis and brachial blood pressure was measured. New comorbidities included systemic infection, congestive heart failure and trauma that occurred within 1 month or less. Cardiovascular disease (CVD) was recorded too. Dialysis adequacy and residual renal function were calculated by a standard technique. RESULTS: Based on SGA, 15.6% of our CAPD patients were malnourished. The malnourished patients had advanced age, higher CRP and ECW/TBW levels than normally nourished patients (age: 68.78 +/- 11.92 vs. 59.26 +/- 13.46 years, p = 0.001; CRP: 11.98 +/- 20.22 vs. 5.56 +/- 8.30 mg/l, p = 0.004; ECW/TBW: 0.55 +/- 0.16 vs. 0.52 +/- 0.04, p = 0.049). Patients with malnutrition were more prone to have CVD (53.13 vs. 31.79%, p = 0.004) and new comorbidities (65.62 vs. 4.62%, p = 0.023). Multivariate analysis showed new comorbidities, mostly systemic infection, which were associated with nutritional status (p < 0.001). Both ECW/TBW and new comorbidities were associated with serum CRP, CVD and malnutrition (p < 0.001-0.05). In contrast, some traditional factors which were recognized as contributing to malnutrition such as residual renal function, dialysis adequacy, metabolic acidosis, total protein loss, diabetes and Charlson indexes were not different between normally nourished and malnourished patients in the present study. CONCLUSIONS: Our results suggest that only 15.6% of patients were malnourished in our PD program. Old age, inflammation, CVD, fluid overload and new comorbidities were all associated with malnutrition, with new comorbidities, mostly systemic infections, being the most significant risk factor. However, many traditional factors such as residual renal function, dialysis adequacy and diabetes were not. PMID- 17077623 TI - Evidence that C-reactive protein or IL-6 are not surrogates for all inflammatory cardiovascular risk factors in hemodialysis patients. AB - BACKGROUND/AIMS: In otherwise healthy adults, high C-reactive protein (CRP) levels are associated with cardiovascular disease and have been linked to an inflammatory state. The presence of vascular disease is also associated with increased expression of adhesion molecules, including soluble intercellular adhesion molecule (sICAM), vascular endothelial growth factor (VEGF) and leukocyte-derived myeloperoxidase (MPO). These associations suggest potential mechanisms whereby inflammation may injure the vascular endothelium, but the recognition of how these mediators act in concert remain poorly characterized. That the prevalence of atherosclerosis and markers of inflammation are increased in renal failure patients suggests that inflammation causes accelerated vascular disease. METHODS: In hemodialysis patients, we examined the relationships between plasma CRP and sICAM, VEGF and MPO longitudinally. We determined whether episodes of a high CRP value were paralleled by simultaneous increases in mediators of inflammatory injury or molecules associated with endothelial cell adhesion or growth and whether CRP levels correlated with those of VEGF and MPO. RESULTS: Episodic increases in CRP were accompanied by higher levels of VEGF, sICAM and MPO. However, there was no correlation between serum CRP levels or other acute phase proteins and either MPO or VEGF, nor was there a constant temporal relationship between MPO and CRP. By contrast, MPO and VEGF levels were closely correlated with one another during episodes of inflammation (p = 0.0001), and CRP and interleukin-6 levels were also correlated. Increases in MPO tended to be restricted to patients with grafts or catheters, and not those with AV fistulas. CONCLUSIONS: These results suggest that high plasma levels of CRP or other acute phase proteins in cross-sectional studies should be interpreted cautiously when defining mechanisms underlying cardiovascular disease in the hemodialysis patient population. One, or more than one inflammatory repertoire may be activated, one involving hepatic acute phase proteins and the other neutrophil activation and each may contribute separately to outcomes. Better prognostic information may be obtained by measurement of more markers than CRP alone, such as MPO and VEGF. PMID- 17077625 TI - Sequential hemoperfusion and continuous venovenous hemofiltration in treatment of severe tetramine poisoning. AB - OBJECTIVE: It was the aim of this study to observe the effects of sequential hemoperfusion (HP) and continuous venovenous hemofiltration (CVVH) on patients with severe tetramine poisoning and to evaluate the ability of these modalities to remove tetramine. METHODS: Eighteen patients diagnosed as having severe tetramine poisoning were treated by blood purification, additional to routine medical therapy. Blood purification procedures included HP using activated charcoal for 3-5 h and consecutive CVVH for 24-36 h. Patients' clinical conditions, blood routine tests and serum chemical tests were evaluated every day after admission. Plasma tetramine concentrations were determined before and after HP. During CVVH, tetramine concentrations in plasma before and after passing through the filter and ultrafiltration at 2 and 12 h were also determined. RESULTS: Eight patients received blood purification within 12 h after onset of poisoning, and 10 patients received blood purification more than 12 h later. Early-treated patients showed a higher cure rate (100 vs. 60.0%; p < 0.05, chi(2) test) and shorter coma time than late-treated patients (26.0 +/- 23.2 h, range 5 70, vs. 59.7 +/- 27.7 h, range 20-96; p < 0.01, rank test). The mean plasma tetramine concentrations in early- and late-treated patients were comparable (0.095 +/- 0.036 vs. 0.134 +/- 0.110 mg/l; p > 0.05). Mean plasma tetramine concentration was reduced from 0.124 +/- 0.082 to 0.080 +/- 0.055 mg/l after HP. At 2 h of CVVH, mean plasma tetramine concentration was 0.078 +/- 0.064 mg/l, at 12 h of CVVH, 0.074 +/- 0.059 mg/l, and the ultrafiltration sieving coefficient at 2 and 12 h was 0.839 +/- 0.409 and 0.686 +/- 0.253 mg/l, respectively. CONCLUSION: Early sequential HP and CVVH therapy may significantly improve the outcome of patients with severe tetramine intoxication. HP can rapidly reduce the plasma concentration of tetramine, and CVVH can attenuate the plasma tetramine concentration rebound after HP by continuously removing tetramine from the plasma. PMID- 17077626 TI - The effect of intravenous iron on oxidative stress in hemodialysis patients at various levels of vitamin C. AB - BACKGROUND/AIMS: Vitamin C levels decrease during hemodialysis (HD), which deteriorates antioxidant defense. Vitamin C may also act pro-oxidatively, via reduction in Fe(III). We sought to determine whether intravenous iron (Fe(iv)) induced oxidative stress differs in HD patients with low and physiological vitamin C levels and whether intravenous vitamin C (C(iv)) administration during HD would change the response to Fe(iv). PATIENTS AND METHODS: Twenty patients with vitamin C deficiency (median 15.7 micromol/l, range 8.0-22.7) received Fe(iv) (100 mg iron sucrose between 150 and 180 min of HD). After 4 weeks of oral supplementation, the levels of vitamin C were comparable with those of controls (60.1 micromol/l, range 47.4-70.9). Patients were subsequently treated with (1) Fe(iv), (2) Fe(iv) and continuous 2 mg/min C(iv) throughout HD, (3) saline (S), and (4) S+C(iv). Plasma thiobarbituric acid reacting substances (TBARS) and vitamin C were assessed before, during and after FE(iv)(S), and 15, 30 and 60 min after infusion. RESULTS: Fe(iv) induced a comparable rise in TBARS in patients with vitamin C deficiency (before Fe(iv), 1.9 micromol/l, range 1.4-1.9; after Fe(iv), 2.6 micromol/l, range 2.3-2.9; p < 0.01) and in those with normal vitamin C (before Fe(iv), 1.9 micromol/l, range 1.7-2.1; after Fe(iv), 2.6 micromol/l, range 2.5-2.9; p < 0.01). Fe(iv)+C(iv) resulted in a greater increase in TBARS (after Fe(iv), 3.1 micromol/l, range 2.8-3.2) compared with Fe(iv) (p < 0.01). CONCLUSION: Iron sucrose-induced oxidative stress is comparable in HD patients with vitamin C deficiency and in those with normal vitamin C. We documented a pro oxidative effect of vitamin C during Fe(iv)+C(iv) administration. PMID- 17077627 TI - Digitalis treatment decreases mortality and morbidity in heart failure patients. Reanalysis of digitalis treatment. PMID- 17077628 TI - Navel piercing as a cause for Streptococcus viridans endocarditis: case report, review of the literature and implications for antibiotic prophylaxis. AB - We describe a case of Streptococcus viridans endocarditis in a 17-year-old female with congenital ventricular septal defect, that followed shortly after performing naval piercing. She was eventually treated with penicillin with complete recovery. We review other cases from the literature of piercing-associated endocarditis, and suggest that individuals at particularly high risk might be considered for antibiotic prophylaxis prior to such procedures. PMID- 17077629 TI - Non-compaction on autopsy in Duchenne muscular dystrophy. AB - Left ventricular hypertrabeculation (LVHT)/non-compaction is frequently associated with neuromuscular disorders. Recently, LVHT has been detected in a 28 year patient with Duchenne muscular dystrophy. Here, the patho-anatomic findings of this patient are presented, which showed LVHT located within in the apex and the anterior and lateral wall, being the most demanded segments during systole. The septum and the left ventricular outflow tract were not involved. The patho anatomic specimen also showed aberrant bands and false tendons, a frequent finding in hearts with LVHT. The patho-anatomic findings were in line with those of LVHT patients with or without neuromuscular disorders. PMID- 17077630 TI - Pioglitazone improves endothelial function in non-diabetic patients with coronary artery disease. AB - OBJECTIVE: To test the hypothesis that pioglitazone, a peroxisome proliferator activated receptor-gamma agonist, will improve endothelial function in non diabetic subjects with coronary artery disease, we conducted a prospective study to evaluate the effect of this medication on the brachial artery vasomotor function and circulating markers of endothelial activation. METHODS: Baseline characteristics were collected. After initial endothelial function assessment, patients were treated with pioglitazone hydrochloride 30 mg daily. The medication was continued for 12 weeks and endothelial function was reassessed as well as the inflammatory markers. The study medication then was stopped, and all the tests were repeated 12 weeks later. RESULTS: Seventeen subjects completed all three study phases. Mean age was 58 (range: 36-77 years). Compared with the baseline, the endothelium-dependent vasodilation improved significantly with the treatment (p < 0.001) from 4.4 +/- 3.9 to 8.4 +/- 4.1%, a relative increase of 91%. After withdrawal of treatment, the endothelium-dependent vasodilation returned towards baseline values. There was no change in endothelium-independent vasodilatation (12.27 +/- 6.35 to 13.9 +/- 9.23%, to 12.42 +/- 5.35%, p = 0.177). The urine asymmetric dimethlyarginine levels decreased significantly with the treatment, but also returned to the initial values after the wash-out period (1.27 +/- 0.5 micromol/ml to 0.97 +/- 0.3 micromol/ml to 1.34 +/- 0.5 micromol/ml, p = 0.017). No difference in the lipid profile, C-reactive protein, erythrocyte sedimentation rate, or fibrinogen levels was seen. CONCLUSION: Pioglitazone rapidly improves endothelial function in non-diabetic patients with coronary artery disease. This improvement is associated with a change in mean urinary asymmetric dimethylarginine levels, although a cause and effect cannot be determined from this investigation. PMID- 17077631 TI - Agitation in Dutch institutionalized patients with dementia: factor analysis of the Dutch version of the Cohen-Mansfield Agitation Inventory. AB - BACKGROUND/AIMS: To establish the construct validity of the Dutch version of the Cohen-Mansfield Agitation Inventory (CMAI-D) in institutionalized patients with dementia. METHODS: The CMAI-D was administered to a large sample of 1,437 patients with moderate to severe dementia, receiving nursing home or outreaching nursing home care. Exploratory factor analysis was used to examine the behavioral dimensions underlying CMAI-D observations. RESULTS: A restricted 3-factor solution showed 3 factors, i.e. physical aggression, physically nonaggressive behavior and verbally agitated behavior, with prevalences of 62, 67 and 62%, respectively. An unrestricted factor solution revealed 3 additional behavioral dimensions: hiding/hoarding, vocal agitation and a factor of miscellaneous items (i.e. repetitious mannerisms, spitting), which occurred in 30, 28 and 35% of the patients, respectively. CONCLUSION: The 3-factor solution of physical aggression, physically nonaggressive behavior and verbally agitated behavior corroborates earlier findings in other patient samples and therefore establishes the construct validity in institutionalized patients with severe dementia. The robustness of these findings across different care settings suggests that agitated behaviors have a common basis. In addition, unrestricted factor analysis showed 3 other important independent behavioral symptoms in dementia, but they are in fact too small to be used as a subscale. These findings might add to the taxonomy of agitation and aggression in dementia. PMID- 17077632 TI - Apolipoprotein E genotype influences presence and severity of delusions and aggressive behavior in Alzheimer disease. AB - AIM: We investigated differences in the prevalence and severity of 10 neuropsychiatric and behavioral symptoms according to apolipoprotein E (APOE) genotype and dementia severity in Alzheimer disease (AD). METHODS: Neuropsychiatric and behavioral symptoms of 110 AD patients were assessed using the Neuropsychatric Inventory. Dementia severity was assessed using the Mini Mental State Examination (MMSE). RESULTS: There were 27 APOE-epsilon4-negative patients, 65 heterozygous patients and 18 homozygous patients. There was a significant association between the number of APOE epsilon4 alleles and prevalence and severity of neuropsychiatric and behavioral symptoms that was mainly attributable to delusions and agitation/aggression, which were more common and severer among homozygous APOE epsilon4 carriers. In addition, the presence of hallucinations, anxiety, apathy and aberrant motor behavior increased with deteriorating MMSE score, independently of APOE epsilon4 status. CONCLUSIONS: The present study showed that the APOE epsilon4 genotype modifies neuropsychiatric and behavioral phenotype in AD. In particular, it was shown that delusions and agitation/aggression were more common and severer among homozygous APOE epsilon4 carriers than among heterozygous or APOE-epsilon4-negative patients. PMID- 17077633 TI - Intracranial pressure parameters in idiopathic normal pressure hydrocephalus patients with or without improvement of cognitive function after shunt treatment. AB - BACKGROUND/AIMS: Previous research has shown improvements in the clinical triad (gait, incontinence and cognitive dysfunction) after shunt surgery in idiopathic normal pressure hydrocephalus (iNPH) patients with intracranial pulse pressure amplitudes >4-5 mm Hg, the pulse amplitudes being <4 mm Hg in the nonresponders. However, it is unknown whether similar differences exist regarding change in cognitive functioning after shunt surgery. The aim of this study was to compare preoperative intracranial pressure (ICP) parameters between iNPH that either improved or not improved in cognitive function after shunt treatment. METHODS: Neuropsychological testing was performed before and after surgery in 27 consecutive iNPH patients treated with ventriculoperitoneal shunts. The ICP recordings were performed as part of routine preoperative assessment, stored as raw data files, and analyzed retrospectively. The mean ICP as well as single ICP wave amplitudes were computed and analyzed in consecutive 6-second time windows. RESULTS: Significant improvement in neuropsychological tests, defined as a 4 point improvement in Folstein Mini-Mental State Examination or improvement by one standard deviation in 50% of subtests of Dementia Rating Scale, was found in 12 patients (44%; shunt responders). In these patients, mean ICP was similar though the mean ICP wave amplitude was significantly higher than in the shunt nonresponders. CONCLUSIONS: While preoperative mean ICP was similar, the mean ICP wave amplitudes were considerably higher in iNPH patients with significant change in cognitive function following shunt surgery. PMID- 17077634 TI - An adult case of megalencephalic leukoencephalopathy with subcortical cysts with S93L mutation in MLC1 gene: a case report and diffusion MRI. PMID- 17077635 TI - Isolated vomiting due to a unilateral dorsal vagal complex lesion. PMID- 17077636 TI - Change of dialect after stroke: a variant of foreign accent syndrome. PMID- 17077637 TI - Sequelae of thallium poisoning: clinical and neurophysiological follow-up. PMID- 17077638 TI - Postirradiation spinal root cavernoma. PMID- 17077639 TI - Multiple system atrophy with progressive nocturnal hypoxemia: case report with polysomnography and continuous positive airway pressure treatment. PMID- 17077640 TI - Neuroimage: monomelic amyotrophy. PMID- 17077641 TI - Louis Pierre Gratiolet (1815-1865): the cerebral lobes and fissures. AB - Early anatomists referred to parts of the brain as 'posterior' and 'anterior.' But after comparing many primate brains, Gratiolet noticed a pattern of folds or sulci and convolutions marked by fissures of increasing complexity in the more advanced mammalian brains. The primary fissures he could also identify in the smoother ape brains. He was the first to separate four brain lobes (frontal, parietal, temporosphenoidal, occipital) and a fifth central lobe or insula. He demonstrated a sequence of closure of the cranial sutures, and he identified and named the optic radiation. Because he was reluctant to equate brain size with intelligence, Gratiolet was involved in a prolonged polemic with Broca. PMID- 17077642 TI - Linkage disequilibrium and haplotype homozygosity in population samples genotyped at a high marker density. AB - OBJECTIVE: Analyze the information contained in homozygous haplotypes detected with high density genotyping. METHODS: We analyze the genotypes of approximately 2,500 markers on chr 22 in 12 population samples, each including 200 individuals. We develop a measure of disequilibrium based on haplotype homozygosity and an algorithm to identify genomic segments characterized by non-random homozygosity (NRH), taking into account allele frequencies, missing data, genotyping error, and linkage disequilibrium. RESULTS: We show how our measure of linkage disequilibrium based on homozygosity leads to results comparable to those of R(2), as well as the importance of correcting for small sample variation when evaluating D'. We observe that the regions that harbor NRH segments tend to be consistent across populations, are gene rich, and are characterized by lower recombination. CONCLUSIONS: It is crucial to take into account LD patterns when interpreting long stretches of homozygous markers. PMID- 17077643 TI - Regulation of Wolffian duct development. AB - Wolffian ducts (WDs) are the embryonic structures that form the male internal genitalia. These ducts develop in both the male and female embryo. However, in the female they subsequently regress, whereas in the male they are stabilised by testosterone. The WDs then develop into separate but contiguous organs, the epididymis, vas deferens and seminal vesicles. Recently, considerable progress has been made in identifying genes that are involved in these different stages of development which is described in this review. In addition, WD development in (atypical forms of) cystic fibrosis and intersex disorders, such as the complete androgen insensitivity syndrome, 17beta-hydroxysteroid dehydrogenase deficiency and LH-receptor defects, is discussed. The apparent increase in male reproductive tract disorders is briefly discussed from the perspective of the potential endocrine-disrupting effects of the numerous chemicals in the environment to which the developing male foetus can be exposed. PMID- 17077644 TI - Inhibition of IL-4/IL-13 does not enhance the efficacy of allergen immunotherapy in murine allergic airway inflammation. AB - BACKGROUND: Successful allergen-specific immunotherapy (SIT) is associated with reduced Th2 cytokine production and the induction of IL-10-producing regulatory T cells. To improve treatment efficacy, we investigated the impact of an IL-4/IL-13 inhibitor during SIT. METHODS: BALB/c mice were sensitized intranasally with ovalbumin (OVA) for 4 weeks. Subsequently, they were subjected to intranasal SIT, with OVA being administered at doses increasing from 1 mug to 1 mg over 3 weeks with or without an IL-4/IL-13 inhibitor. Serum OVA-specific antibodies were measured and bronchoalveolar lavage (BAL) fluids were checked for airway eosinophilia. Subsequently, lung tissue was examined histologically for inflammatory infiltrates. Cytokines were detected in BAL fluids and spleen cell cultures. Furthermore, CD4 CD25 double-positive spleen T cells were checked for intracellular IL-10 production by flow cytometry. RESULTS: OVA sensitization resulted in persistent IgE synthesis and an eosinophil-rich allergic airway inflammation combined with increased IL-4 and IL-5 levels. Therefore, intranasal SIT could efficiently reverse the allergic phenotype. This was associated with decreased IL-4 and IL-5 levels, and increased IL-10 levels in BAL fluids as well as increased amounts of IL-10-producing CD25+ regulatory T cells. However, mice treated with the IL-4/IL-13 inhibitor during SIT did not produce significantly different results . CONCLUSION: The use of an IL-4/IL-13 inhibitor as an adjuvant for SIT did not enhance anti-allergic effects. Thus, the observed reversal of Th2 responses during SIT may not be the keystone for successful therapy, but rather other factors, e.g. IL-10-producing regulatory T cells, may be crucial. PMID- 17077645 TI - Interleukin-6 genetic ablation protects from trinitrobenzene sulfonic acid induced colitis in mice. Putative effect of antiinflammatory cytokines. AB - BACKGROUND: Interleukin (IL)-6 is a proinflammatory cytokine implicated in the pathogenesis of inflammatory bowel disease. IL-6 is locally upregulated in inflammatory bowel disease patients and in murine models of experimental colitis. Treatment with anti-IL-6 receptor antibody ameliorates intestinal inflammation. OBJECTIVE: It was the aim of this study to investigate the role of genetic IL-6 deficiency in trinitrobenzene sulfonic acid (TNBS)-mediated colitis, an experimental model inflammation that shares several features with Crohn's disease in humans. METHODS: Acute colitis was induced in wild-type and IL-6-deficient (Il 6(-/-)) mice by intracolonic administration of TNBS. Forty-eight hours after treatment, the local and systemic features of inflammation, i.e. food intake, weight loss, histological markers of colitis, cytokine expression by real-time PCR, food intake and body weight changes, were assessed. RESULTS: In wild-type mice, TNBS administration increased both IL-6 serum levels and local expression of IL-6 by 36 and 9 fold, respectively, compared with control, vehicle-injected mice. Compared with the wild-type mice, the Il-6(-/-) mice had significantly reduced intestinal inflammation as evidenced by epithelial damage, neutrophil infiltration, colon thickness and proinflammatory cytokine expression, following treatment with TNBS. Moreover, baseline levels of the antiinflammatory cytokines IL-10 and tumor growth factor-beta were significantly elevated in Il-6(-/ )compared with the wild-type mice. CONCLUSIONS: Our results demonstrate that Il 6(-/-)are partially protected from the development of TNBS-induced acute experimental colitis, most likely via their significantly elevated baseline levels of antiinflammatory cytokines. PMID- 17077646 TI - Rapid endocytosis of copper-zinc superoxide dismutase into human endothelial cells: role for its vascular activity. AB - Cytosolic CuZn-SOD (SOD1) is a dimeric, carbohydrate-free enzyme with a molecular weight of about 32 kDa and also circulates in human blood plasma. Due to its molecular mass it has been believed that the enzyme cannot penetrate the cell membrane. Here we report that rapid endocytosis of FITC-CuZn-SOD into human endothelial cells occurs within 5 min. Moreover, relaxation of rat aortic rings in response to CuZn-SOD is associated with a lag time of 45-60 s and only observed in the presence of intact endothelial cells. The results indicate acute and rapid endothelial cell endocytosis of CuZn-SOD, possibly via activation of a receptor-mediated pathway. Intracellular uptake via endocytosis may contribute to the vascular effects of CuZn-SOD, including vasodilation, and is likely to play a role in regulation of vascular tone and diseases such as atherosclerosis. PMID- 17077647 TI - Airway smooth muscle relaxations induced by dipyrone. AB - BACKGROUND: Dipyrone differs from other nonsteroidal anti-inflammatory drugs with a distinctive spasmolytic effect; however, the mechanism of action is not clear yet. The possible mechanism behind this effect was investigated on airway smooth muscle tone in vitro. METHOD: The effect of dipyrone on inositol phosphate (IP) accumulation was evaluated on guinea pig trachea smooth muscle. Changes in intracellular free calcium levels and IP accumulation were evaluated in LTK8 cells. RESULTS: Dipyrone (0.01, 0.1, 1 mmol/l) had a relaxing effect on histamine (0.02 mmol/l)- and adenosine triphosphate- (ATP) (0.01 mol/l)-induced contractions, but not potassium chloride (KCl)-stimulated contractions. This relaxing effect was not observed with indomethacin. Indomethacin did not inhibit the relaxation response of dipyrone. In isolated tracheal smooth muscle, histamine- (0.02 mmol/l) and ATP (0.01 mmol/l)-induced IP accumulation was significantly inhibited by dipyrone (1 mmol/l). ATP (0.01 mmol/l)-induced IP accumulation was also significantly inhibited by dipyrone (0.01 mmol/l) in LTK8 cells. Dipyrone inhibited ATP-induced (0.01 and 0.1 mmol/l) intracellular calcium increase in LTK8 cells. CONCLUSION: Inhibition of the release of intracellular Ca(2+) may play a role in the smooth muscle relaxing effect of dipyrone. The inhibitor effect of dipyrone on IP accumulation may be due to direct inhibition of phospholipase C (PLC) or impairment of the activation of PLC by G protein coupled receptor (GPCR). PMID- 17077648 TI - Developmental and behavioral consequences of prenatal fluoxetine. AB - BACKGROUND: Fluoxetine, a selective serotonin reuptake inhibitor, is the most commonly prescribed antidepressant drug for pregnant women. Studies regarding the teratogenic effect of fluoxetine on human and animal models are mainly concerned with structural malformation (congenital anomalies). AIM: Hence, the present study was planned to evaluate the postnatal behavioral effects of fluoxetine on albino rats. METHODS: Three groups of female rats received either distilled water or doses of fluoxetine 8 and 12 mg/kg orally from the 6th to the 20th day of pregnancy. Weaning of the pups was done on the 21st day followed by a battery of behavioral tests to assess for any behavioral effect. The tests included negative geotaxis, open field exploration, rota-rod test, elevated plus maze and passive avoidance test. RESULTS: In the present study there was no change in the gestational length of pregnancy, no premature birth or miscarriage during pregnancy. A high dose of in utero fluoxetine resulted in a decrease in birth weight of the offspring and also reduced weight gain during the preweaning period. No major congenital abnormalities were observed in the offspring exposed to fluoxetine. Prenatal fluoxetine exposure at high dose caused an initial transient delay in motor development and this poor motor activity was transient and not permanent. However, prenatal exposure to fluoxetine at a higher dose showed a favorable effect on learning and memory in water maze and passive avoidance tests. CONCLUSIONS: From the present study, it may be concluded that prenatal fluoxetine causes a transient delay in motor development but does not adversely affect the postnatal behavioral consequences. PMID- 17077649 TI - Direct anatomical localization of the subthalamic nucleus on CT with comparison to Schaltenbrand-Wahren atlas. AB - Stimulation of the subthalamic nucleus (STN) has become an accepted treatment for motor symptoms of Parkinson's disease. Generally, localization of the target is based on stereotactic atlas coordinates and magnetic resonance (MR) images. In this paper a method of direct anatomical localization of the STN on computed tomography (CT) images is described and compared with the Schaltenbrand-Wahren atlas. Serial CT scans were obtained at 1.5-mm interval and scan thickness of 3 mm, with the CT gantry and glabella-inion plane of the patient's head vertical. The STN was identified as a dense structure medial to the inner semicircular border of the internal capsule in the subthalamic region. Its visibility was increased by changing the window and level settings on the CT workstation computer. The center of the STN in relationship to the intercommissural line, determined by this technique, was similar to those found on standard atlases. PMID- 17077650 TI - A simple stereotactic method for frameless deep brain stimulation. AB - BACKGROUND: Deep brain stimulation (DBS) is widely used to treat advanced Parkinson's disease, other movement and psychiatric disorders. DBS implantation requires application of a stereotactic frame throughout a lengthy procedure, making it uncomfortable and tiring. We designed a stereotactic cube to stage the operation, perform frameless microelectrode recording (MER) and fix the DBS. METHODS: The 15-mm cube is implanted in a burr hole using bone cement. It contains 5 parallel trajectories (central + 4 around). It is aligned by stereotactic frame so that central trajectory reaches the target. Frameless MER is performed by attaching a micro-driver to the cube using 2-5 cannulae (4 cm). The DBS is fixed to the cube by a mini-plate and 1 screw. Ninety-six cubes were compared with 43 Bennet spheres (BS). RESULTS: No cube moved compared to 2 (5%) BS (p < 0.05). The final trajectory was central in 64.4% of cubes compared to 47.5% of BS, and the final target was >2 mm out in no cubes compared to 12.5% of BS (p < 0.01). Infection and haemorrhage were observed in 2.5% and 3.3% of cubes, respectively, while 5% of BS developed infection, 5% haemorrhage and 7.5% skin erosion. CONCLUSIONS: This method is simple and effective in staging DBS procedures, performing frameless MER and DBS implantation, fixation and revision. PMID- 17077651 TI - Gamma knife radiosurgery in the treatment of choroidal neovascularization (wet type macular degeneration). AB - We evaluated retrospectively our institutional experience in the treatment of macular degeneration with gamma knife radiosurgery (GKR). Treatment was delivered in a single shot of 12 Gy. Seven patients were treated between March of 1999 and May of 2000. The median duration of follow-up was 2.2 years. The majority of patients maintained stable visual acuity after treatment. Our series indicates that GKR may be useful as a salvage treatment for patients who have failed or are ineligible for other treatments for their macular degeneration. Further studies are needed to better define the role of GKR in the treatment of macular degeneration. PMID- 17077652 TI - Ophthalmic cyclosporine use in ocular GVHD. AB - PURPOSE: To evaluate the safety and efficacy of topical ophthalmic cyclosporine in patients with dry eyes secondary to graft-versus-host disease (GVHD). METHODS: Records of consecutive patients with ocular GVHD treated with topical cyclosporine were reviewed. RESULTS: Thirty-two eyes of 16 patients were included with a mean follow-up of 90 days. Dry eye symptoms improved in 10 patients (62.5%), and corneal fluorescein staining improved in all eyes (P = 0.0039). Global physician impression was favorable in 12 patients (75%). Ocular burning or irritation was noted in 3 patients (19%), which improved with either continuation of cyclosporine at the current dosing regimen or reduction in dose and/or concentration of cyclosporine. CONCLUSION: Ophthalmic cyclosporine seems to be safe and may be beneficial for patients with ocular GVHD, particularly those with more severe dry eyes. PMID- 17077653 TI - Changes in corneal thickness in patients with treated and untreated ocular hypertension. AB - PURPOSE: The elevated intraocular pressure (IOP) in eyes with ocular hypertension is often accompanied by increased corneal thickness. We tested the hypotheses that chronically elevated IOP causes a slow increase in corneal thickness and that lowering the IOP reverses this slow increase. METHODS: Fifty patients with ocular hypertension were randomized to medication and observation groups in the Mayo Clinic site of the Ocular Hypertension Treatment Study. Central corneal thickness was measured using an optical pachymeter at baseline and annually for 6 years. The rates of change of corneal thickness was compared between the groups. Epithelial thickness was measured by confocal microscopy 8 years after the baseline examination. RESULTS: Corneal thickness increased 1.5 +/- 3.3 microm/yr in the observation group (n = 23) and decreased -1.3 +/- 2.8 microm/yr in the medication group (n = 27, P = 0.002). Both rates were significantly different from zero (P = 0.04 and P = 0.02, respectively). Epithelial thickness was 46.4 +/ 4.9 microm in the observation group and 41.3 +/- 4.4 microm in the medication group (P = 0.008). CONCLUSION: Results of this single-center series imply that corneal thickness increases slowly in eyes with ocular hypertension and decreases slowly if the IOP is lowered by topical medications. These phenomena could be explained by a causal relationship between elevated IOP and a slow increase in corneal thickness. A decrease in epithelial thickness accounts for a portion of the thinning that occurs with treatment. If confirmed in a larger series, these findings indicate that the effects of previous treatment on thickness should be considered if corneal thickness is to be used as a discriminant factor in the management of patients with ocular hypertension. PMID- 17077654 TI - Efficacy of a new warm moist air device on tear functions of patients with simple meibomian gland dysfunction. AB - PURPOSE: To evaluate the safety and efficacy of an original warm moist air device on tear functions and ocular surface of patients with simple meibomian gland dysfunction (MGD). METHODS: Fifteen patients with simple MGD and 20 healthy volunteers were recruited in an initial prospective interventional clinical trial to evaluate the safety and short-term effects of the warm moist air device. The device was applied to the eyes of the subjects for 10 minutes. Temperatures of the eye lids and corneas were measured with an infrared thermometer. Symptoms of ocular fatigue were scored using visual analog scales (VASs). Schirmer test, tear film break-up time (BUT), DR-1 tear film lipid layer interferometry, fluorescein staining, and rose bengal staining were also performed before and after the application of the eye steamer. After the initial study, another 2-week prospective clinical trial was carried out in 10 patients with MGD who received the warm moist air treatment. Ten other patients were also recruited and received warm compress treatment with hot towels for 2 weeks to evaluate the long-term effects of the warm moist air device and the warm compresses on tear film lipid layer thickness and ocular surface health. The warm moist air device and the warm compresses were applied for 10 minutes twice a day. The changes in VAS scores for symptoms, BUT values, fluorescein, and rose bengal staining scores were examined before and after each treatment during the second trial. RESULTS: VAS scores of ocular fatigue improved significantly with short- and long-term applications of the warm moist air device in both studies. The mean corneal surface and eye lid temperatures showed significant elevation within safe limits 10 minutes after the moist air application. The mean BUT prolonged significantly in the patients receiving warm moist air applications but did not change significantly in those treated with warm compresses. DR-1 tear film lipid layer interference showed evidence of lipid expression in the patients and controls, with thickening of the tear film lipid layer after 10 minutes of warm moist air device use. In the 2 week trial, tear film lipid layer thickness increased in both warm moist air device and warm compress groups, with a greater extent of increase in the warm moist air device group. CONCLUSION: Warm moist air device use provided symptomatic relief of ocular fatigue and improvement of tear stability in patients with MGD. The new warm moist air device seems to be a safe and promising alternative in the treatment of MGD. PMID- 17077655 TI - Association between meibomian gland changes and aging, sex, or tear function. AB - PURPOSE: To study changes in the lid margin and meibomian glands and their association with aging, sex, and tear function. METHODS: We examined 354 eyes in 177 subjects (76 men and 101 women; 21-93 years; mean age, 63.0 +/- 14.3 years) with no ocular symptoms or ocular surface disorders. Anatomic changes in the lid margin were studied using slit-lamp biomicroscopy. Meibomian gland function and morphology were evaluated on the basis of meibum expression and meibography, respectively. Tear function and ocular surface epithelium were assessed with the Schirmer test, by tear film break-up time, and with a fluorescein staining test. RESULTS: Eyes with abnormal lid margin anatomy, hyposecretion of meibum, and meibomian gland dropout were seen in 26 (7.3%), 46 (12.4%), and 68 eyes (18.6%), respectively, with a significant association between each finding and aging (P = <0.0001, 0.0498, and <0.0001, respectively). In patients < or =69 years of age, no significant association was found between meibomian gland-related findings and sex. However, a high incidence of abnormal lid margin and gland dropout was noted in men > or =70 years of age compared with women. No significant association was found between changes in the lid margin and meibomian glands and tear function in patients > or =40 years of age. CONCLUSION: Among symptom-free subjects, we found that changes in the lid margin and meibomian glands were closely related to aging. Among elderly subjects, changes in the anatomic lid margin and meibomian gland morphology were observed more frequently in men than in women. Tear function showed no association with either changes in the lid margin or function of the meibomian glands. PMID- 17077656 TI - Keratoprosthesis: current techniques. AB - PURPOSE: To compare and contrast the techniques and results of keratoprosthesis (KPro) using the Cardona device (1975-1985) with those related to the use of the Dohlman-Doane type 1 device (2003-2005). METHODS: We conducted a retrospective review of 31 cases of Cardona KPro implanted between 1975 and 1981 and a review of 25 cases of Dohlman-Doane type 1 KPro implanted between 2003 and 2005. Procedures were limited to eyes with a poor prognosis for traditional penetrating keratoplasty. RESULTS: In the Cardona cases, 39% (12/31) of patients achieved a visual acuity of 20/40 or better at some point postoperatively, degrading to 16% (5/31) at the end of the observation period. Fifty percent (15/31) required subsequent surgical revision of the KPro. Sixteen percent of cases (5/31) developed endophthalmitis, and 58% (18/31) developed retroprosthetic membranes. There were 5 dislocations of the prosthesis and an additional 3 frank extrusions (26%). Of the 25 Dohlman-Doane cases, there have been no reoperations, endophthalmitis, dislocations, or extrusions to date (8/15/05). Forty-eight percent (12/25) achieved 20/200 acuity or better, and 12% (3/25) achieved 20/40 or better. Retroprosthetic membranes formed in 3 cases. CONCLUSION: Although KPro remains a technique to be used in poor prognosis keratoplasty, improvements in the design of the device and surgical technique over the past 30 years have reduced the overall morbidity and severity of complications and improved the prognosis for visual rehabilitation. PMID- 17077657 TI - Immunologic and clinical manifestations of infectious scleritis after pterygium excision. AB - PURPOSE: To present the immunologic status and clinical manifestations of patients who had infectious scleritis after pterygium excision. METHODS: This prospective noncomparative study of immunologic status and clinical manifestations involved 18 eyes of 18 patients with infectious scleritis (16 bacterial and 2 fungal infections) with a history of pterygium excision from 1999 to 2001. RESULTS: The period between pterygium excision and scleritis ranged from 1 to 36 years. None of the 18 patients had any history of systemic autoimmune disease. Elevated erythrocyte sedimentation rates were found in 9 (50%) of 18 patients. Elevated serum C-reactive protein levels were found in 9 (75%) of 12 patients. Other serologic tests were all negative. The most common pathogen of infectious scleritis, Pseudomonas aeruginosa, was present in 13 patients (72.2%), significantly higher than any others (P < 0.05). All of the eyeballs were salvaged, and 9 (50%) of 18 patients had final vision better than 2/20. CONCLUSION: There was no underlying autoimmune disease associated with infectious scleritis in this study. Early diagnosis and aggressive medical and surgical treatment might have saved the eyes. PMID- 17077658 TI - Mooren ulcer: an immunopathologic study. AB - PURPOSE: To determine the pattern and distribution of mononuclear cells, adhesion, and co-stimulatory molecules in the conjunctiva of patients with Mooren ulcer. METHODS: Conjunctival biopsy specimens were obtained from 6 patients with Mooren ulcer and 6 healthy individuals. Immunohistochemistry was performed on frozen sections of the cryopreserved human conjunctivas using monoclonal antibodies directed against CD1alpha, CD3, CD4, CD8, CD20, CD25, CD57, and CD68 cells; the adhesion molecules E-selectin, vascular cell adhesion molecule-1 (VCAM 1), very late activation-4 (VLA-4), ICAM-1, and LFA-1; and the co-stimulatory molecules CD28, B7-1, B7-2, and CTLA-4. RESULTS: Differences in expression on the conjunctival epithelium from patients with Mooren ulcer and normal subjects were noted only for VCAM-1, VLA-4, ICAM-1, and LFA-1. The ratio of CD4+/CD8+ cells in Mooren ulcer specimens was significantly higher (3.5-fold). However, in the substantia propria, Mooren ulcer specimens revealed significantly increased numbers of CD1alpha+, CD3+, CD4+, CD20+, CD28+, B7-1+, B7-2+, and CD68+ cells. The ratios of CD4+/CD8+ cells and B7-2+/antigen-presenting cells in Mooren ulcer specimens were significantly higher (5-fold). All tested adhesion molecules showed significant up-regulation in the patients' conjunctivas. Mooren ulcer vascular endothelial cells prominently expressed E-selectin, VCAM-1, VLA-4, and ICAM-1 compared with normal conjunctiva. CONCLUSION: The simultaneous presence of multiple types of inflammatory cells, adhesion, and co-stimulatory molecules in Mooren ulcer conjunctiva suggests that their interaction may contribute to a sustained immune activation as at least part of the pathogenic mechanism of this disorder. PMID- 17077659 TI - Efficacy of topical cyclosporine 0.05% in the treatment of dry eye associated with graft versus host disease. AB - PURPOSE: To assess the efficacy of topical cyclosporine 0.05% (Restasis) in patients with dry eye associated with graft versus host disease after stem cell transplantation. METHODS: After completing a 3-month run-in period of using only artificial tears to control dry eye symptoms in both eyes, patients who failed to achieve adequate relief (n = 8) were instructed to instill topical cyclosporine twice a day. Visual acuity, slit-lamp appearance, and intraocular pressure were evaluated every 2 weeks for a minimum of 3 months. In addition, Schirmer basal secretion tests, noninvasive fluorescein breakup time, and tear lysozyme were also performed. Patients were also given a dry eye questionnaire regarding symptoms of burning, tearing, and blurred vision. RESULTS: Dry eye signs improved significantly with cyclosporine treatment in 7 of 8 patients. Cyclosporine provided statistically significant improvements in Schirmer basal secretion scores (P = 0.003), tear breakup time (P = 0.002), and tear lysozyme levels (P = 0.033) after 3 months of treatment. CONCLUSION: The findings in this prospective study suggest that dry eye associated with graft versus host disease can be effectively treated with topical cyclosporine, especially in patients unresponsive to other treatment modalities. These findings should be further evaluated in large-scale, controlled clinical trials. PMID- 17077660 TI - Validity of fluorexon disodium versus sodium fluorescein for use in Goldmann tonometry. AB - PURPOSE: To evaluate the safety, validity, and comfort of 0.35% fluorexon disodium and 0.4% benoxinate (Flura-Safe) compared with the gold standard of 0.25% sodium fluorescein and 0.4% benoxinate for Goldmann applanation tonometry (GAT). METHODS: This was a double-masked, randomized, crossover clinical trial. Subjects received either the standard or study formulation for GAT on visit 1 and the other formulation 1 week later. At each visit, tonometer mire quality, adequacy of fluorescence, ease of intraocular pressure (IOP) measurements, the IOP value, and anesthetizing efficacy of the formulation were assessed. Subjects graded general comfort, soreness and irritation, and burning and stinging of each formulation at 1 and 5 minutes after drop instillation. RESULTS: Sixty-seven subjects completed the study. The mean IOP was 13.9 +/- 2.7 with fluorexon and 13.9 +/- 2.8 mm Hg with fluorescein OD and 14.0 +/- 2.8 with fluorexon and 13.9 +/- 2.5 mm Hg with fluorescein OS. The measurements with the 2 formulations were highly correlated for OD and OS, and the differences between the 2 measurements were not clinically significant. There was also no significant difference between the 2 drops in mire clarity, adequacy of fluorescence, or corneal anesthesia. However, fluorexon was statistically more comfortable (P = 0.039) and caused less stinging and burning (P = 0.014) at 1 minute versus the fluorescein formulation. CONCLUSION: Not only was the new fluorexon product accurate and effective in GAT, it was also statistically more comfortable and had a less stinging and burning effect at 1 minute after drop instillation than the traditional fluorescein formulation. Because fluorexon is less likely to stain soft contact lenses, this may be the dye-anesthetic formulation of choice for practices that routinely perform GAT. PMID- 17077661 TI - Oral cimetidine adjuvant therapy for recalcitrant, diffuse conjunctival papillomatosis. AB - PURPOSE: To describe a case of recurrent, advanced conjunctival papillomatosis, treated by oral cimetidine (CIM) combined with secondary surgical intervention and an application of intraoperative mitomycin C. METHODS: Case report and literature review. RESULTS: A 9-year-old boy suffered from recurrent, progressive, diffuse multifocal conjunctival papillomatosis over the left upper and lower palpebral and the fornical conjunctiva. He underwent 3 separate surgeries; however, they did not prevent tumor recurrence. The recurrent lesions were more severe and extensive than before the surgeries. To avoid postoperative symblepharon, ankyloblepharon, dry eye, and possible corneal neovascularization after extensive lesion excision, oral CIM at a dosage of 200 mg 4 times daily was administered for 4 months before surgery. A debulking excision of the residual tumor with an intraoperative application of mitomycin C was performed as a secondary therapy after the main mass decreased in size. Postoperative oral CIM was continued for 6 months. The papillomatosis cleared without recurrence or symblepharon, ankyloblepharon, conjunctival scarring, or corneal neovascularization after 4 years of follow-up examinations. CONCLUSION: Oral CIM can be used as an initial, nonsurgical strategy for treating cases of massive, recalcitrant conjunctival papillomatosis. If there is tumor shrinkage, surgical debulking with applications of mitomycin C may be sufficient to eliminate any residual tumor tissue without inducing conjunctival scarring or corneal neovascularization. PMID- 17077662 TI - Astigmatism after mersilene and nylon suture use for penetrating keratoplasty. AB - PURPOSE: To evaluate postoperative astigmatism after the use of Mersilene and nylon sutures for penetrating keratoplasty. METHODS: Two groups of 12 patients each (age, 20-50 years) suffering from keratoconus underwent penetrating keratoplasty. Continuous suturing combined with 12 interrupted sutures was used in each case: Mersilene 11-0 sutures were used in group 1, whereas nylon 10-0 sutures were used in group 2. Sutures were removed to control astigmatism when required. All patients were monitored for 36 months. RESULTS: During the follow up period, sutures were removed in each eye as indicated by astigmatism or breakage. In group 1 (Mersilene 11-0 sutures), no breakage of sutures was recorded. In group 2 (nylon 10-0 sutures), breakage of sutures was recorded in 11 of 12 cases. However, continuous sutures were removed in 2 patients in group 2. After the first year, the astigmatism remained within +/-1.00 diopter in all except 1 patient in group 1 and in 5 patients in group 2. No suture-related complications were recorded except for scarring with photophobia in 1 patient in group 1 and suture breakage in 11 eyes in group 2. CONCLUSION: Mersilene sutures can provide a stable postoperative astigmatism, whereas nylon sutures have a tendency to break and cause changes in corneal astigmatism. PMID- 17077663 TI - Reliability of grading lissamine green conjunctival staining. AB - PURPOSE: To assess the reliability of a lissamine green grading scale for conjunctival images. METHODS: A 20-second video clip of the right eye of 288 contact lens-wearing individuals was recorded using a digital slip-lamp camera after instilling liquid lissamine green. A single nasal and temporal still image were selected. A masked reader used the Oxford grading scale to grade the images on two occasions whereas a second masked reader graded each image on 1 occasion. kappa statistics and 95% confidence intervals (CIs) were used to determine the within- and between-grader reliability overall and when the sample was stratified by age, sex, contact lens type, and disease severity. RESULTS: There was substantial within-grader reliability for both the nasal (kappasimple = 0.69, 95% CI, 0.63-0.75) and temporal (kappasimple = 0.73, 95% CI, 0.67-0.79) images. There was moderate between-grader reliability for both the nasal (kappasimple = 0.51, 95% CI, 0.44-0.58) and temporal (kappasimple = 0.51, 95% CI, 0.44-0.58) images. Age, sex, and contact lens type did not affect within- or between-examiner reliability. There may have been an influence of disease severity on within examiner reliability, because grading of the temporal images was significantly less reliable in the images with more significant staining. CONCLUSION: Within- and between-grader reliability of lissamine green staining seems to be at least substantial to moderate. Because the extent of conjunctival staining may influence reliability, this should be considered when studies may include patients with significant staining. PMID- 17077664 TI - Ulcerative keratitis caused by Haemophilus influenzae. AB - PURPOSE: To report the clinical course, treatments, antibiotic sensitivities, and visual outcomes for eyes with ulcerative keratitis caused by Haemophlilus influenzae. METHODS: The medical records of 10 patients with culture-proven H. influenzae-associated corneal ulcer in a tertiary referral center in a 5-year period (1998-2003) were retrospectively reviewed. RESULTS: The median age was 61 years (range, 2-83 years). The mean follow-up time was 9.4 months (range, 2-31 months). Risk factors such as previous surgeries (5), herpes simplex keratitis (2), leukoma adherens with calcification (1), and exposure keratitis (1) were identified in 9 patients. In vitro testing of the H. influenzae isolates showed that 6 of 10 were resistant to ampicillin; all were sensitive to amoxicillin/clavulanic acid, ceftriaxone, and cefuroxime. All cases responded well to medical antibiotic treatment. Final best spectacle-corrected visual acuity ranged from no light perception to 20/30. CONCLUSION: H. influenzae is a rare cause of corneal ulceration. Predisposing factors for infection are usually present. Accurate diagnosis and treatment may preserve ocular integrity and visual acuity. PMID- 17077665 TI - Technique for preparation of the corneal endothelium-Descemet membrane complex for transplantation. AB - PURPOSE: Replacing diseased corneal endothelium with a preparation of Descemet membrane carrying functional endothelium and no stroma may be a feasible method for treating corneal endothelial decompensation. To obtain a viable donor of a Descemet membrane endothelium disc, we modified the Descemet membrane stripping technique and monitored the percentage of endothelial damage to the donor tissue preparation. METHODS: Forty-eight human corneas were used. Cornea buttons were mounted on an artificial anterior chamber, endothelial side up. Endothelia were stained with alizarin red, examined under the microscope, and photographed at 5 different sites (microscope, x100; digital magnification, x2.83). A 6 x 7-mm rectangular piece of endothelium-Descemet membrane complex was obtained using a Grieshaber microsurgical knife and Kelman-McPherson forceps. Digital photographs of endothelia were analyzed with a computer, and the percentage of endothelial damage was calculated. Specimens were processed for hematoxylin-eosin staining. RESULTS: Forty of 48 endothelium-Descemet membrane preparations (83.3%) were complete peels with minimal endothelial damage. Endothelial damage before and after the surgery was 1.57 +/- 2.11% and 2.61 +/- 1.77%, respectively. Eight preparations (16.7%) failed because of tearing. Multiple hematoxylin-eosin stained sections showed the presence of endothelium with intact Descemet membrane and no stromal tissue. CONCLUSION: We modified the technique of Melles and obtained a sheet of Descemet membrane and endothelium with minimal endothelial damage and with no remaining stroma observed. This simple technique can be used to obtain the endothelium-Descemet membrane complex in minutes. It may be useful for corneal endothelium transplantation. PMID- 17077666 TI - Inhibition of inflammatory cytokine production in human corneal cells by dexamethasone, but not cyclosporin. AB - PURPOSE: To evaluate the modulatory effects of anti-inflammatory agents, dexamethasone (Dex) and cyclosporin A (CsA), on the production of cytokines and chemokines by human corneal cells in vitro following stimulation by the pro inflammatory cytokine after interleukin 1beta (IL-1beta). METHODS: A human corneal epithelial (HCE) cell line and human corneal fibroblasts (HCFs) were stimulated in culture with IL-1beta and treated with Dex or CsA. The gene expression for selected cytokines and chemokines was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). The secretion of cytokines and chemokines was measured by enzyme-linked immunosorbent assay. RESULTS: IL-1beta enhanced the mRNA and/or protein levels of granulocyte-macrophage colony stimulating factor (GM-CSF), IL-8, and monocyte chemotactic protein (MCP)-1 in HCE and IL-6, IL-8, MCP-3, and regulated on T-cell activation expressed secreted (RANTES) in HCFs. Treatment with CsA did not inhibit cytokine production in either HCE or HCFs. In contrast, Dex treatment inhibited the IL-1beta-induced production of GM-CSF, IL-6, IL-8, MCP-3, and RANTES, but not MCP-1. CONCLUSION: These results show that Dex, but not CsA, has direct immunosuppressive effects on the resident corneal cells, HCE and HCFs. This suggests that the clinically observed immunosuppressive effects of topical CsA are mediated primarily through the immune cells. PMID- 17077667 TI - Ocular surface epithelial cells up-regulate HLA-G when expanded in vitro on amniotic membrane substrates. AB - PURPOSE: To study the modulation of immunoregulatory genes in ocular surface epithelial cells cultured on amniotic membrane (AM). METHODS: Microarray analysis was performed in a conjunctival epithelial cell line (CCL20.2) expanded on denuded AM. Among the genes that were upregulated by an AM substrate compared with collagen-coated dishes, the fetal nonclassic major histocompatibility complex molecule, HLA-G, was found to be the only immunoregulatory gene up regulated by more than 2.5-fold. Because CCL20.2 is contaminated by HeLa cells, expression of HLA-G mRNA was confirmed in primary-cultured limbal (LE) and conjunctival epithelial (CE) cells by reverse transcriptase-polymerase chain reaction (RT-PCR), semiquantitative real-time PCR, immunocytochemistry, and Western blot analysis. A functional assay was performed using an HLA-G transfected K-562 human erythroleukemia cell line. RESULTS: Freshly dissociated limbal epithelial cells express HLA-G mRNA; however, protein levels were low. Western blots and immunocytochemistry showed that both LE and CE cells upregulated the HLA-G protein when cultured on collagen-coated dishes and on AM. HLA-G mRNA levels were significantly higher in CE cultured on AM compared with collagen. Natural killer (NK) cell-induced cell lysis of an HLA class 1-negative K-562 human erythroleukemia cell line was slightly reduced when transfected with LE-derived HLA-G mRNA. CONCLUSION: CE and LE cells express functional HLA-G when expanded ex vivo, which may affect inflammation and immune reaction when transplanted to the ocular surface. PMID- 17077668 TI - Absence of blood and lymphatic vessels in the developing human cornea. AB - PURPOSE: The normal human cornea is devoid of both blood and lymphatic vessels and actively maintains this avascularity (corneal angiogenic privilege). Whether and when corneal angiogenic privilege is achieved during development is unknown. METHODS: This study analyzed whether the cornea is primarily devoid of both blood and lymphatic vessels during intrauterine development or whether secondary regression of pre-existing vessels occurs before delivery. Indirect double immunohistochemistry was performed on 4-microm serial pupil-optic disc sections of paraffin-embedded human eyes stillborn at gestational ages of 17 to 41 weeks with antibodies against von Willebrand factor (vWF; factor VIII-associated antigen) as a panendothelial marker and with antibodies against lymphatic vessel endothelial hyaluronate receptor 1 (LYVE1) as a marker specific for lymphatic vascular endothelium. RESULTS: Human corneas were devoid of both vWF+++/LYVE-1(-) blood vessels and vWF+/LYVE-1+++ lymphatic vessels at all time-points analyzed. In contrast, there were numerous blood and lymphatic vessels detectable in the adjacent conjunctiva. CONCLUSION: The normal human cornea is primarily avascular and devoid of both blood and lymphatic vessels. Corneal angiogenic privilege is already achieved very early during fetal intrauterine development. This suggests early and strong expression of both antiangiogenic and antilymphangiogenic factors in the human cornea during development. PMID- 17077669 TI - Noninvasive diagnosis of epithelial downgrowth after penetrating keratoplasty using immunohistochemical analysis of resected corneal buttons. AB - PURPOSE: To describe a noninvasive technique that can be used to support a clinically suspected diagnosis of epithelial downgrowth in patients who have previously undergone penetrating keratoplasty (PK). METHODS: Previously resected corneal buttons of 14 patients who developed signs suggestive of epithelial downgrowth after PK were examined retrospectively using immunohistochemical analysis to determine if occult epithelial cells were in fact present on the posterior surface of the cornea. Many of these patients were diagnosed with pseudophakic corneal edema or graft failure and had undergone multiple intraocular surgical procedures. In all cases, the initial histopathologic evaluation did not disclose the presence of epithelial cells using light microscopy after standard handling and staining techniques. Fourteen age- and diagnosis-matched control specimens were also examined. RESULTS: Thirteen of 14 specimens (92.9%) exhibited positive staining with anti-keratin AE1/AE3 markers, indicating that epithelial cells were already present on the endothelial surface at the time of the original PK. None of the control specimens displayed positive staining of the retrocorneal surface. CONCLUSION: Retrospective immunohistochemical analysis of corneal specimens can establish the diagnosis of epithelial downgrowth in a noninvasive manner. Early diagnosis may allow for this condition to be treated more successfully if aggressive intervention is undertaken. Alternatively, it may spare the patient from additional (and potentially futile) intraocular surgery. In the clinical setting of suspected epithelial downgrowth, ophthalmic pathologists should re-evaluate PK specimens from patients initially diagnosed with pseudophakic corneal edema or graft failure. PMID- 17077670 TI - Correlation between clinical in vivo confocal microscopic and ex vivo histopathologic findings of Salzmann nodular degeneration. AB - PURPOSE: To report clinical, in vivo confocal microscopy and ex vivo histopathologic findings of Salzmann nodular degeneration (SND). METHODS: A 48 year-old woman with symptoms of ocular irritation and decreased visual acuity caused by SND in both eyes was treated by corneal scraping and phototherapeutic keratectomy (PTK). Slit-lamp biomicroscopy, in vivo confocal microscopy, ex vivo light microscopy, immunohistology, and corneal topography were performed. RESULTS: In vivo confocal microscopy showed an irregular network of highly reflective structures representing activated keratocytes, which could be seen by light microscopy and characterized immunohistologically as myofibroblasts. Unstructured areas with increased reflectivity correlated with irregularly arranged collagen fibers and hyaline deposits in the nodulus. Epithelial cells in vivo appeared atypically shaped and elongated. These observations were consistent with decreased thickness of the epithelium over the nodules showed by histopathology. Treatment led to a dramatic reduction of hyperopia. Two months after surgery, uncorrected visual acuity (UCVA) in the right eye was 20/32 and 20/20 with a refraction of -0.75 -0.75/0 degrees. UCVA in the left eye was 20/40 and 20/20 with a refraction of +0.50 -1.75/165 degrees. Corneal topography showed regular astigmatism. CONCLUSION: In vivo confocal microscopy confirmed the clinicopathologic findings of Salzmann's nodular degeneration. Observations by in vivo confocal microscopy were consistent with the histopathologic descriptions of SND. PMID- 17077671 TI - Acute hydrops in keratoconus masquerading as acute corneal transplant rejection. AB - PURPOSE: To document 2 cases that developed acute hydrops corneae within the host tissue after corneal transplantation. Subsequent extension across the transplant host interface led to edema of the transplanted tissue, with consequent misdiagnosis and treatment of acute transplant rejection. METHODS: The patients attended the Anterior Segment Service at Moorfields Eye Hospital, London, UK. RESULTS: Both patients had a reduction in vision, ocular discomfort, and stromal edema of the transplanted button and adjacent host cornea. No break in the Descemet membrane was noted in either case. CONCLUSION: It is important to differentiate hydrops of the host cornea from acute transplant rejection. This distinction will allow the judicious use of steroids and may allay patient anxiety about possible rejection. PMID- 17077672 TI - "Steel wool keratopathy": a clinical sign of chronic inflammation. AB - PURPOSE: To introduce into the clinical nomenclature a sign frequently observed in our patients with persistent corneal inflammation associated with herpetic stromal keratitis. METHODS: Case reports and review of the literature. RESULTS: Four representative patients with herpesvirus stromal keratitis are presented. Herpes simplex virus-1 (HSV-1) was confirmed by culture in 1 case and by polymerase chain reaction in a second case. In the remaining 2 cases, the diagnosis was made based on characteristic clinical findings for herpes simplex virus and varicella zoster virus (VZV). On clinical examination, all 4 representative cases of stromal keratitis revealed a well-defined, localized region of intertwined, metallic-like, polychromatic material in the corneal stroma, a sign we have termed steel wool keratopathy. We have only rarely observed this finding in patients with stromal keratitis not caused by a herpesvirus. CONCLUSION: Steel wool keratopathy seems to represent a focal region of stromal degeneration or deposition associated with chronic inflammation. Although we most often observe this finding in patients with stromal keratitis secondary to HSV or VZV, we cannot exclude the possibility that this sign represents the sequelae of chronic/recurrent inflammation rather than a specific pathologic response to herpetic antigens. PMID- 17077673 TI - Multifocal squamous cell carcinoma of the conjunctiva with intraocular penetration in a patient with AIDS. AB - PURPOSE: To describe the clinical findings and to study the histopathology of conjunctival masses in a patient with HIV infection. METHODS: A case report. RESULTS: The histopathology from a 38-year-old Thai female patient who presented with visual loss on the left eye was studied. She had been diagnosed with AIDS 3 years before and was treated with highly active antiretroviral therapy and anti tuberculous drugs for 1 year. The left eye was blind and showed 2 separate whitish masses on the bulbar conjunctiva in adjacent to the inferomedial limbus. Multiple nodules under the conjunctiva were seen inferiorly. Computerized tomography of the orbit revealed slight thickening of the left lower eyelid but could not show any globe involvement. The patient underwent orbital exenteration. Gross pathology showed multiple areas of the tumor. Histopathology revealed dysplastic squamous epithelium invading into the substantia propria of the conjunctiva with intraocular penetration. The anterior surface of the iris was filled with tumor cells containing keratin pearls. CONCLUSION: Squamous cell carcinoma can be multifocal and aggressive in HIV-infected patients. Rapid progression to intraocular penetration can be observed. PMID- 17077674 TI - Inadvertent retention of Descemet Membrane in penetrating keratoplasty. AB - PURPOSE: To present a case of inadvertently retained Descemet Membrane (DM). METHODS: Case report. RESULTS: A 41-year-old male with previous history of retinal detachment surgery underwent a repeat penetrating keratoplasty; the first graft decompensated secondary to silicone oil. Postoperatively, an inadvertently retained DM was observed forming a physical barrier to the silicone oil. CONCLUSION: Retained DM is a rare complication of penetrating keratoplasty, with only a few cases reported in the literature. This membrane served a protective role from the effects of silicone oil on the donor cornea but later opacified and required surgical intervention. PMID- 17077675 TI - Corneal topographic features in a family with nanophthalmos. AB - PURPOSE: To characterize the corneal topographic features in a family with nanophthalmos. METHODS: Two observational case reports and a review of the literature. Corneal shape was analyzed using a topographic modeling system (TMS) and an Orbscan topographic system, with further analysis based on Fourier series and Zernicke polynomials. A control group was used for comparison. RESULTS: Two female family members with nanophthalmos belonging to the same generation showed significant irregular corneal astigmatism and corneal steepening. CONCLUSION: Topographic corneal steepening and irregular astigmatism seems to be associated in 1 family with inherited nanophthalmos. Higher degrees of irregular astigmatism, which were not evident without a topographic analysis, may account, in part, for the unexplained visual reduction. PMID- 17077677 TI - Long-term results after bare sclera pterygium resection with excimer smoothing and local application of mitomycin C. PMID- 17077679 TI - Long-term results after bare sclera pterygium resection with excimer smoothing and local application of mitomycin C. PMID- 17077681 TI - Fluid resuscitation and the septic kidney. AB - Acute renal failure is a common complication of sepsis in the critically ill patient. Fluid resuscitation is considered a cornerstone for preservation of function in the septic kidney. This is generally based on the assumption that fluid therapy will restore and maintain adequate renal blood flow. This principle, however, has not yet been formally evaluated in humans. Thus, it remains unclear how fluid therapy may affect renal blood flow in septic acute renal failure. Further, there is new evidence to imply that the choice, timing and amount of fluid used for resuscitation in sepsis may have a direct impact on kidney function. Thus, in this editorial, we consider the relevant literature and more recent insights into the effect of fluid resuscitation on the septic kidney. PMID- 17077682 TI - Acute kidney injury: epidemiology and diagnostic criteria. AB - PURPOSE OF REVIEW: To review recent advances in the definitions and diagnostic criteria for acute renal failure and acute kidney injury. To explore how these changes impact the epidemiology and clinical implications for patients in the intensive care unit. RECENT FINDINGS: Recently published consensus criteria for the definition of acute renal failure/acute kidney injury have led to significant changes in how we think about this disorder. Studies from around the world, both in and out of the intensive care unit, have shown a dramatic incidence of acute kidney injury and high associated mortality. This review considers these new findings and their historical context, and attempts to shed new light on this old problem. SUMMARY: Small changes in kidney function in hospitalized patients are important and impact on outcome. RIFLE criteria provide a uniform definition of acute kidney injury and are increasingly used in literature. PMID- 17077683 TI - The epidemiology of acute renal failure in the world. AB - PURPOSE OF REVIEW: The aim of this article is to describe our current understanding of the epidemiology of acute renal failure, especially in severe cases requiring renal replacement therapy. Some data from an international observational study (the Beginning and Ending Supportive Therapy (BEST) for the kidney study) are also presented. RECENT FINDINGS: Multiple epidemiological studies for acute renal failure have been conducted in different populations using various criteria and reported a wide range of incidence and mortality. In several multicenter studies reporting the incidence of renal replacement therapy requirement in a general intensive care unit population, however, renal replacement therapy requirement and hospital mortality was quite similar among the studies, approximately 4% and 60%, respectively. In North and South America, nephrologists typically manage acute renal failure patients with intermittent renal replacement therapy. On the other hand, in Asia, Australia and Europe, intensivists are commonly responsible for managing these patients with continuous renal replacement therapy. SUMMARY: Although the epidemiology of acute renal failure is varied among regions and centers, the average incidence of renal replacement therapy requirement and hospital mortality in critical illness are similar in multicenter studies. Continuous renal replacement therapy is gaining popularity and intensivists are becoming responsible for managing patients with it. More studies are needed to understand acute renal failure epidemiology worldwide. To conduct such studies, consensus criteria for acute renal failure and a well performed acute renal failure-specific severity score will be required. PMID- 17077684 TI - Epidemiology of renal recovery after acute renal failure. AB - PURPOSE OF REVIEW: Recovery of renal function after acute renal failure is an important clinical determinant of patient morbidity. Herein, the epidemiology of renal recovery after acute renal failure will be described, along with potential predictive factors and interventions. RECENT FINDINGS: Renal recovery has been variably defined, most often as recovery to independence from renal replacement therapy. A recent consensus definition for acute renal failure has been published and included provisions for defining renal recovery. Renal recovery to renal replacement therapy independence occurs in the majority by hospital discharge and peaks by 90 days. All of older age, female sex, co-morbid illnesses, especially chronic kidney disease, and late initiation of renal replacement therapy or conventional intermittent renal replacement therapy have been coupled with non recovery. Analysis of the literature suggests several interventions may influence recovery. SUMMARY: The prognosis is generally good for recovery after acute renal failure. Most patients will be independent of renal replacement therapy by 90 days. Additional research is necessary, however, to understand recovery rates not only to independence from renal replacement therapy, but also to complete and partial recovery. Future studies need to consider the health economic implications for survival and non-recovery. Finally, questions on the role of various interventions require characterization in randomized controlled trials to determine how they may influence renal prognosis. PMID- 17077685 TI - Current worldwide practice of dialysis dose prescription in acute renal failure. AB - PURPOSE OF REVIEW: The practice of renal replacement therapy for acute renal failure varies greatly around the world. This is especially true for the dose of renal replacement therapy. This review describes the epidemiology of dialysis prescription and delivery around the world based on recent observational studies and international surveys. Furthermore, a brief speculation on the concept of adequacy of therapy is provided. RECENT FINDINGS: Following the publication of the first large prospective randomized trial on the effects of treatment dose on outcome of patients with acute renal failure a certain consensus has been achieved on the adequacy of 35 ml/kg/h of clearance in continuous therapies. Surveys conducted in 1998 had demonstrated that such adequate therapy was only prescribed in a minority of patients. Recent observations based on questionnaires and surveys confirm that the number of centers prescribing adequate dialysis dose is increasing, but there is still a large number of institutions where prescription is made with no specific adequacy targets and effective delivery is not measured. Several barriers have been identified to reach adequacy targets including logistics, costs, personnel and others. SUMMARY: The analysis of the epidemiology of dialysis dose around the world shows a complex picture. Wide variations in practice are present although a significant effort has been made to standardize the prescription and delivery of therapy. PMID- 17077686 TI - The epidemiology of acute renal failure: 1975 versus 2005. AB - PURPOSE OF REVIEW: Data from the mid 1970s relating to the clinical features, pathogenesis, treatment and outcome of patients affected by acute renal failure were compared with data reported in the literature in 2005. RECENT FINDINGS: The differences in patient characteristics over the last 30 years are profound and claims that the outcome of acute renal failure has not changed over time are equally misleading. Thirty years later patients are older, have more comorbidities, develop renal failure in association with interventions (liver transplantation, lung transplantation, heart and lung transplantation) that essentially did not exist in the 1970s, are sicker with more complex symptoms, receive more effective therapies and survive at higher rates once outcome is adjusted for illness severity. SUMMARY: The treatment of critically ill patients with acute renal failure has changed dramatically over the last 30 years. As treatment has increased in efficacy and sophistication, so has the severity of the condition. PMID- 17077687 TI - The long-term outcome after acute renal failure. AB - PURPOSE OF REVIEW: Research priorities in critical care are increasingly focusing on long-term outcomes and prognosis for survivors of critical illness. This review will focus on long-term outcomes after acute renal failure. RECENT FINDINGS: Few studies have described the long-term outcomes after acute renal failure. Rates of survival are variable and range from 46 to 74%, 55 to 73%, 57 to 65% and 65 to 70% at 90 days, 6 months, 1 year and 5 years, respectively. All of older age, co-morbid illness, illness severity, septic shock, and renal replacement therapy after cardiac surgery have been associated with reduced survival. Recovery to independence from renal replacement therapy is expected in 60-70% of survivors by 90 days. Health-related quality of life is generally good and perceived as acceptable. Survivors often experience difficulty with mobility and limitations in activities of daily living. Renal replacement therapy is costly and achieves marginal cost-effectiveness in terms of quality-adjusted survival for those with a higher probability of survival. SUMMARY: The long-term survival after acute renal failure is poor. Yet, most survivors recover sufficient function to become independent from renal replacement therapy. While perceived health-related quality of life is good, survivors have a lower health related quality of life compared with the general population. Further research is needed to explore the relationship between survival, markers of morbidity and costs after acute renal failure. PMID- 17077689 TI - Measurement of acid-base resuscitation endpoints: lactate, base deficit, bicarbonate or what? AB - PURPOSE OF REVIEW: Inadequate oxygen delivery to the tissues frequently results in significant metabolic acidosis. The resultant cellular and organ dysfunction can increase morbidity, mortality and hospital stay. Early diagnosis of shock can lead to early resuscitation efforts that can prevent ongoing tissue injury. This review focuses on the metabolic, hemodynamic and regional perfusion endpoints utilized in the diagnosis of metabolic acidosis resulting from shock. Resuscitation strategies aimed at supranormal oxygen delivery will be discussed. RECENT FINDINGS: Serum pH, lactate, base deficit and bicarbonate have all been extensively studied as clinical markers of metabolic acidosis in shock. While their trend helps guide resuscitation, no single marker or specific value can be utilized to guide resuscitation for all patients. Hemodynamic parameters and regional tissue endpoints are designed to identify compensated shock before it progresses to uncompensated shock. Resuscitation strategies initiated in the early phases of shock can reduce complications and death. Efforts to resuscitate patients to supranormal oxygen delivery endpoints have demonstrated mixed success, with several notable complications. SUMMARY: Despite the large number of endpoints available to the clinician, none are universally applicable and none have independently demonstrated improved survival when guiding resuscitation. Patients who respond well to initial resuscitation efforts demonstrate a survival advantage over nonresponders. PMID- 17077690 TI - Can we safely restrict transfusion in trauma patients? AB - PURPOSE OF REVIEW: Transfusion of red blood cells in the trauma patient can be lifesaving. The question is how much and when? It is important to weigh the risks and benefits of red blood cell transfusions, as well alternatives to transfusion as these products are not benign. RECENT FINDINGS: We explore the evidence, and provide the rationale for current and future red blood cell transfusion strategies within a framework of prehospital and hospital care of the trauma patient. We also describe how red blood cell transfusion trends are changing in trauma, discuss alternatives to red blood cell transfusion and present evidence from randomized controlled trials that support a lower transfusion trigger. SUMMARY: Optimal transfusion practice and use of alternatives in trauma is a rapidly expanding and important area of research. Strong clinical evidence derived by future randomized controlled trials in the area of transfusion triggers as well as transfusion alternatives is required to determine their roles in clinical practice. PMID- 17077691 TI - Role of thoracoscopy in acute management of chest injury. AB - PURPOSE OF REVIEW: To review the literature on the use of video-assisted thoracoscopic surgery for the diagnosis and treatment of intrathoracic injuries. RECENT FINDINGS: Video-assisted thoracoscopic surgery is a relatively recent innovation. It was originally promoted for the treatment of retained hemothorax and the diagnosis of diaphragm injury. It is highly effective for the management of those problems. Recent studies have focused on video-assisted thoracoscopic surgery for treatment of chest wall bleeding, diagnosis of transmediastinal injuries, pericardial window and persistent pneumothorax. In properly selected patients, video-assisted thoracoscopic surgery is extremely efficacious in managing these problems. SUMMARY: The role of video-assisted thoracoscopic surgery in the management of acute chest injury is expanding. It is an invaluable tool for the trauma surgeon. PMID- 17077692 TI - Failure of nonoperative management of abdominal solid organ injuries. AB - PURPOSE OF REVIEW: Nonoperative management of the spleen has been the conventional approach for dealing with blunt splenic injury in children for 25 years. Following acceptance in the field of pediatric surgery, nonoperative management of blunt injury to the liver and spleen became the template in adult trauma surgery. It has proven to be of unequivocal benefit to the majority of hemodynamically stable pediatric and adult patients who have suffered blunt liver or splenic trauma. Offsetting these gains, has been the presence of failures. RECENT FINDINGS: The recent literature has focused on factors which may impact the nonoperative management success or failure rate. These factors include initiation of guidelines, risk of overwhelming postsplenectomy infection, character of clinical judgment, role of computed tomography in detecting associated intraabdominal injuries, the presence of more than one solid organ injury, risk of associated hollow viscus injury, and the drawbacks of angioembolization. SUMMARY: Despite the failures of nonoperative management outlined in this review, the approach has been generally successful. Efforts at improving organ salvage rates and diminishing failures with this approach continue. Notwithstanding our enthusiasm to advance this method of patient care, we must avoid imperiling a subpopulation of patients in our attempt to improve nonoperative management success rates. PMID- 17077694 TI - Management and infrastructure in critical care: a priority? PMID- 17077693 TI - The impact of endogenous triggers on trauma-associated inflammation. AB - PURPOSE OF REVIEW: Inflammation immediately starting after trauma is a consequence of an efficient host defense system that is not only capable of sensing exogenous and pathogen-derived danger signals, but also endogenous, multifunctional alarm signals, which both can initiate an inflammatory response. RECENT FINDINGS: Even in the absence of infection, Toll-like receptors play an important role in inflammation via recognition of host-derived, endogenous 'damage signals' like heat shock proteins and 'alarmins' such as the nuclear protein high-mobility group box protein 1, which are presented as a result of tissue trauma. In addition to the Toll-like receptors, a number of other receptors are involved in the host inflammatory response, including the new family of nucleotide oligomerization domain-like receptors capable of sensing the presence of danger signals in the cytoplasm. Important links occur between the Toll-like receptors as key inducers of the pro-forms of interleukin-1beta and interleukin-18 and the activation of certain nucleotide oligomerization domain like receptors, resulting in inflammasome formation--an essential process leading to the secretion of these proinflammatory cytokines. SUMMARY: In addition to improved insights into the regulation of traumatic inflammation and the etiology of the systemic inflammatory response syndrome, some endogenous immune triggers seem to have the potential to serve as novel biomarkers in predicting post traumatic complications. PMID- 17077695 TI - Team care: beyond open and closed intensive care units. AB - PURPOSE OF REVIEW: Evidence supporting dedicated intensivist staffing in intensive care units is growing. Despite clinical and economic benefits, medical staff politics and a shortage of intensivists impede the intensivist model. The purpose of this paper is to accelerate patient's exposure to the benefits of intensivists, and introduce team care in the intensive care unit. RECENT FINDINGS: The cost savings achieved through intensivist staffing range from $510,000 to $3.3 million. The intensivist model may only have been adopted by 4% of intensive care units. Barriers to implementing the model are shortage of intensivists, reimbursement for intensivists, and political will. Four attributes make the model ideal: physical presence, knowledge of critical care practice, coordination of team care, and unit management. It may be helpful to not label intensive care units as open or closed and consider team care, whereby hospitals seek to achieve the attributes of the model given their resources and culture. SUMMARY: Intensivists save lives and costs. By working toward team care, hospitals may achieve a successful intensivist model, and patients may realize the benefits of spending less for healthcare and living longer. To achieve this model, physician and hospital leaders must form a partnership. PMID- 17077696 TI - The evolving role of the acute care nurse practitioner in critical care. AB - PURPOSE OF REVIEW: The newest nurse practitioner role is the acute care nurse practitioner. This paper presents the latest data on the role from both a US and international perspective. RECENT FINDINGS: Now present in the USA for at least 15 years, the acute care nurse practitioner role has become well established in critical care settings and is moving into international healthcare. The few outcome studies conducted to date demonstrate the acute care nurse practitioner provides quality patient and family care, improves patient satisfaction, is cost effective, and is an answer to the hospital's shortage of medical residents with new restrictions on working hours. SUMMARY: The role of acute care nurse practitioners in critical care is increasing worldwide. Most countries are experimenting with this latest nurse practitioner as an extended-role healthcare provider with many potential benefits to patients and their families, as well as the healthcare system. PMID- 17077697 TI - The perspective of families of the critically ill patient: their needs. AB - PURPOSE OF REVIEW: An appreciation of a family's difficult experience in dealing with their critically ill loved ones has created the concept of 'family-centered care'. The purpose of this paper is to review the literature on the needs of the family during their intensive care unit experience. RECENT FINDINGS: Families consistently highlight three major issues that they deem could be improved from their perspective. Increased information about their loved ones, proximity to the patient and a more flexible visiting policy stand out as relevant issues to families. SUMMARY: Medical staff and administrators should recognize that families of critically ill patients have particular needs that help them cope with having their loved ones in an intensive care unit. Simple changes in philosophy and policy would greatly decrease the anxiety these families experience. PMID- 17077698 TI - Ethical, legal and organizational issues in the ICU: prediction of outcome. AB - PURPOSE OF REVIEW: Since the development of the first general outcome prediction models, these instruments have been widely used in the intensive care unit. Last updated in the early 1990s, these models are now severely outdated. RECENT FINDINGS: In recent months, researchers and users assisted in several attempts at improving the existing models through customization or expansion or in the development of new models, such as the Simplified Acute Physiology Score (SAPS) 3 and the Acute Physiology and Chronic Health Evaluation (APACHE) IV. SUMMARY: Although not similar, especially in the choice of the reference population, these models aim at replacing older general outcome models, the predictions from which no longer reflect the current case-mix outcomes of intensive care. The objective of this review is to present and discuss, to the clinician working in the intensive care unit, these different strategies and to give an updated version of the general outcome prediction models available in 2006. PMID- 17077699 TI - Bibliography. Current world literature. Renal system. PMID- 17077700 TI - Aligning internal organizational factors with a service excellence mission: an exploratory investigation in health care. AB - In today's competitive health care environment, service excellence is rapidly becoming a major differentiating advantage between health care providers. Too often, senior executives talk about their commitment to a mission statement that extols the virtues of providing world class service to their patients only to undermine those statements with what they do, write, and say. This article presents an exploratory investigation into a new application of an internal mission alignment instrument that seeks to assess the extent to which an organization's internal processes are aligned with its service mission. This instrument was sent to 250 randomly selected employees from all clinical departments of a large southeastern hospital to explore the underlying alignment factors. A factor analysis of the data revealed eight factors that predicted beneficial employee outcomes such as organizational commitment and satisfaction with the job and organization. PMID- 17077701 TI - Using foresight methods to anticipate future threats: the case of disease management. AB - We describe a unique foresight framework for health care managers to use in longer-term planning. This framework uses scenario-building to envision plausible alternate futures of the U.S. health care system and links those broad futures to business-model-specific "load-bearing" assumptions. Because the framework we describe simultaneously addresses very broad and very specific issues, it can be easily applied to a broad range of health care issues by using the broad framework and business-specific assumptions for the particular case at hand. We illustrate this method using the case of disease management, pointing out that although the industry continues to grow rapidly, its future also contains great uncertainties. PMID- 17077702 TI - Using path analysis to examine causal relationships among balanced scorecard performance indicators for general hospitals: the case of a public hospital system in Taiwan. AB - Examining whether the causal relationships among the performance indicators of the balanced scorecard (BSC) framework exist in hospitals is the aim of this article. Data were collected from all twenty-one general hospitals in a public hospital system and their supervising agency for the 3-year period, 2000-2002. The results of the path analyses identified significant causal relationships among four perspectives in the BSC model. We also verified the relationships among indicators within each perspective, some of which varied as time changed. We conclude that hospital administrators can use path analysis to help them identify and manage leading indicators when adopting the BSC model. However, they should also validate causal relationships between leading and lagging indicators periodically because the management environment changes constantly. PMID- 17077703 TI - Antecedents of clinical information technology sophistication in hospitals. AB - Grounded in the resource-based theory and the innovation diffusion theory, this article develops and tests a research model for assessing the antecedents of hospital innovativeness with regard to clinical information technology (IT) applications. A cross-sectional survey was conducted in a sample of U.S. hospitals (n = 74) to assess three dimensions of clinical IT sophistication. Secondary data were used to measure the antecedents, namely, four groups of organizational capacity variables. Bivariate and regression analyses were conducted to identify significant associations. A significant percentage (45-61%) of the variance in clinical IT sophistication was explained, mostly by leadership and knowledge sharing capacities. In particular, IT tenure and technical knowledge resources were significantly related to clinical IT sophistication. Surprisingly, managerial tenure and hospital's belonging to a network showed significant negative associations with two dimensions of the clinical IT sophistication construct. To address the challenges they face, hospitals should consider encouraging career development for current individuals in charge of IT activities, and attracting professionals with an IT background who have the knowledge and ability to trigger new ideas and favor the adoption and use of clinical IT applications in these settings. PMID- 17077704 TI - Can organizational support for technology influence job satisfaction? AB - The relationship between organizational support for telemedicine and nurses' job satisfaction in home health agencies (HHAs) is investigated in this study. Drawing from the theory of perceived organizational support, we propose that HHAs that provide strong support for telemedicine will have higher nurse job satisfaction levels than those that do not. Primary data were collected from 29 HHAs and 917 nurses from 2004 to 2005. The results of the analysis indicate a strong, positive, significant association between organizational support for telemedicine and home health nurses' job satisfaction. We conclude that technological implementation, with sufficient organizational support, can be a strong competitive strategy to attract and retain high-quality nurses. PMID- 17077705 TI - The hospitalist movement in the United States: agency and common agency issues. AB - Agency relationships and their effect on patient representation in the hospitalist role are examined in this article. Emphasis is on relationship(s) with patients, primary care physicians, health plans, and hospitals. PMID- 17077706 TI - Benefiting from networks by occupying central positions: an empirical study of the Taiwan health care industry. AB - At issue is whether network resources imply some resources available to all members in networks or available only to those occupying structurally central positions in networks. In this article, two conceptual models, the additive and interaction models of the firm, are empirically tested regarding the impact of hospital resources, network resources, and centrality on hospital performance in the Taiwan health care industry. The results demonstrate that: (1) in the additive model, hospital resources and centrality independently affect performance, whereas network resources do not; and (2) no evidence supports the interaction effect of centrality and resources on performance. Based on our findings in Taiwanese practices, the extent to which the resources are acquired externally from networks, we suggest that while adopting interorganizational strategies, hospitals should clearly identify those important resources that reside in-house and those transferred from network partners. How hospitals access resources from central positions is more important than what network resources can hospitals acquire from networks. Hospitals should improve performance by exploiting its in-house resources rather than obtaining network resources externally. In addition, hospitals should not only invest in hospital resources for better performance but should also move to central positions in networks to benefit from collaborations. PMID- 17077707 TI - Hospital perceived value. AB - The creation, distribution and communication of value have been considered to be the key element of marketing (American Marketing Association, 2004, www.marketingpower.com). The aim of this article is to identify the indicators of perceived value in a hospital context. The results show that perceived quality and emotions are key dimensions of perceived value. PMID- 17077708 TI - Connection, regulation, and care plan innovation: a case study of four nursing homes. AB - We describe how connections among nursing home staff impact the care planning process using a complexity science framework. We completed six-month case studies of four nursing homes. Field observations (n = 274), shadowing encounters (n = 69), and in-depth interviews (n = 122) of 390 staff at all levels were conducted. Qualitative analysis produced a conceptual/thematic description and complexity science concepts were used to produce conceptual insights. We observed that greater levels of staff connection were associated with higher care plan specificity and innovation. Connection of the frontline nursing staff was crucial for (1) implementation of the formal care plan and (2) spontaneous informal care planning responsive to changing resident needs. Although regulations could theoretically improve cognitive diversity and information flow in care planning, we observed instances of regulatory oversight resulting in less specific care plans and abandonment of an effective care planning process. Interventions which improve staff connectedness may improve resident outcomes. PMID- 17077709 TI - Analyzing administrative costs in hospitals. AB - Using an integrated, management-based approach, this article presents a conceptual framework for analyzing administrative costs in hospitals; the framework differentiates among three types of administrative costs-operational, payer-based, and regulatory--on the basis of the locus of control. An illustration of the framework uses 2003 data for Florida hospitals. PMID- 17077712 TI - Keeping Patients Safe: a collaboration among nurse administrators and researchers. AB - Translating the recommendations from the Institute of Medicine's landmark report, Keeping Patients Safe: Transforming the Work Environment of Nurses, into practice and healthcare policy will require an extensive collaboration among nurse administrators and nurse researchers. The insights and skills of both groups are critical for evaluating the recommendations and their implications for practice, for filling gaps in research, and for influencing significant health system change. PMID- 17077713 TI - Hospital restructuring and nursing leadership: a journey from research question to research program. AB - The 1990s brought new fiscal realities to healthcare, leading to nursing job loss estimates in tens of thousands following widespread hospital restructuring to manage costs and improve efficiency. This research aimed at examining (a) how multiple episodes of hospital restructuring leading to layoff of nurses affected nurses who remained employed and (b) whether and how nursing leadership mitigated or intensified the negative effects of hospital restructuring on nurses. This dissertation comprised 3 empirical studies leading to 5 publications. The first study was a systematic literature review; the second and third used structural equation modeling to develop and test theoretical models addressing nursing practice environments and effects of hospital restructuring on nurses. The combined findings in this dissertation illustrate that hospital restructuring had significant negative physical/emotional health effects on nurses who remained employed. Nurses who worked for resonant (emotionally intelligent) leadership reported positive health and well-being, and opportunities to provide quality patient care. Nurses who worked for dissonant leadership reported greater negative effects of hospital restructuring. These findings led to a beginning theory of relational energy--a mechanism of mitigation whereby resonant nursing leaders invest energy into collaborative relationships with nurses, thereby positively influencing health and well-being, and, ultimately, outcomes for patients. PMID- 17077714 TI - Nurses information-processing patterns: impact on change and innovation. AB - This descriptive study, based on the Organizational Engineering conceptual framework, investigates the information-processing styles of nursing staff (RNs, LPNs, nursing assistants, and unit secretaries) that impact how they accept and deal with change and innovation. The study sample was made up of 578 nursing staff and nurse managers. The major finding is that both nursing staff overall and RNs as a separate group are clustered in the "Conservator" (Logical Process- Hypothetical Analyzer) quadrant of the Organizational Engineering Model. In contrast, the nurse managers were significantly more likely to be in the "Changer" (Reactive Stimulator--Relational Innovator) quadrant. PMID- 17077715 TI - The canaries in the coal mine speak: why someone should (and should not) become a nurse. AB - Nurses in acute care hospitals are like the canaries used to test for lethal gases in mines--if they remain, the conditions must not be extreme. Staff nurses from 8 states responded to a questionnaire giving reasons why someone should and should not be a nurse. The almost 700 nurses responding reported that many of the reasons one should be a nurse are also the reasons one should not be one (eg, pay/benefits, respect for nursing). Extrinsic factors dominated the reasons to be a nurse, with both intrinsic and extrinsic factors being the reasons not to do so. PMID- 17077716 TI - Is patient acuity a proxy for patient characteristics of the AACN Synergy Model for Patient Care? AB - PURPOSE: To test the feasibility of using patient acuity indicators as proxy measures for the AACN Synergy Model for Patient Care patient characteristics. DESIGN AND METHODS: Secondary data analysis of a convenience sample of 481 patient ratings from adult and pediatric critical and noncritical medical surgical patients. FINDINGS: Combining dichotomous patient acuity indicators into multilevel categorical variables allowed evaluation of linear relationships between acuity and patient characteristics indicators. Of the 8 patient characteristics, only participation in care exhibited meaningful correlations with the acuity indicators. According to these findings, acuity indicators should not be substituted for the Synergy Model patient characteristics. PMID- 17077717 TI - The impact of staff nurse empowerment on person-job fit and work engagement/burnout. AB - Workplace empowerment is an important strategy for creating positive nursing work environments in a time of a severe nursing shortage. The purpose of this study was to test a model linking staff nurse perceptions of empowerment to their perceived fit with 6 areas of work life and work engagement/burnout using Kanter's work empowerment theory. We tested the model in a cross-sectional correlational survey design with a random sample of 322 staff nurses in acute care hospitals across Ontario. Overall, staff nurses perceived their work environment to be only somewhat empowering. Fifty-three percent reported severe levels of burnout. Overall empowerment had an indirect effect on emotional exhaustion (burnout) through nurses' perceived fit in 6 areas of work life. The final model fit statistics revealed a good fit (chi2 = 32.4, df = 13, GFI = 0.97, IFI = 0.97, CFI = 0.97, RMSEA = 0.07). These findings have important implications in the current nursing shortage. PMID- 17077718 TI - Barriers and enablers to changing organizational culture in nursing homes. AB - PURPOSE: To discuss the barriers and enablers of changing organizational culture in 3 nursing homes undergoing a culture change initiative, and suggest actions for program enhancement. METHODS: Interview data with staff (n = 64) and families (n = 14) from 3 culture-change facilities in a larger mixed-methods pilot study were used to identify barriers and enablers. FINDINGS: The 3 sites ranged from 120 to 139 beds and did not differ in staff characteristics. Barriers included exclusion of nurses from culture-change activities, perceived corporate emphasis on regulatory compliance and the "bottom line," and high turnover of administrators and caregivers. Enablers included a critical mass of "change champions," shared values and goals, resident/family participation, and empowerment at the facility level. RECOMMENDATIONS: Involve all levels of staff, residents, and community in culture-change activities. Align incentives and rewards with the new values. Empower individual homes to make decisions at the facility level. Work with corporate partners to enable rapid translation and implementation of recommendations based on the findings. PMID- 17077719 TI - Nursing research in leadership/management and the workplace: narrowing the divide. PMID- 17077720 TI - Professional boundary crossings and boundary violations and their implications. PMID- 17077721 TI - Costal cartilage autografts to simulated degenerative intervertebral discs in the rat. AB - STUDY DESIGN: An autograft of costal cartilage was transplanted into the rat intervertebral space in the proximal tail following 2 weeks of simulated degeneration by chondroitinase ABC (CABC). OBJECTIVES: The purpose of this study was to evaluate costal cartilage transplantation into a degenerated disc as a possible therapy. SUMMARY OF BACKGROUND DATA: Reversal of degenerative disc dehydration is an attractive goal. Costal cartilage is plentiful, hydrophilic, and avascular, leading us to speculate that it would survive transplantation into the degenerated disc, increase proteoglycan content, and restore disc height. MATERIALS AND METHODS: Costal cartilage fragments were transplanted into a single proximal intervertebral disc in each of the rats' tails following a 2-week period of simulated degeneration. The intervertebral space was measured on radiographs under 2.5x magnification taken pretreatment and 21 days posttreatment. Each specimen was sagittally sectioned, mounted, and stained. The slides were graded for proteoglycan content. RESULTS: A 64% increase in intervertebral disc height was observed in the implant group compared with a 4% increase in sham operated group and a 39% increase in the CABC only group. Histology demonstrated a viable implant in 7 of 9 rats. The transplant group had significantly more proteoglycan staining than either the CABC group or sham group (P < 0.05). CONCLUSIONS: Costal cartilage transplantation may rehydrate degenerated intervertebral discs and might serve as a promising model for understanding and perhaps modifying this complex degenerative disease. PMID- 17077722 TI - A comparison of pullout strength for pedicle screws of different designs: a study using tapped and untapped pilot holes. AB - STUDY DESIGN: The pullout strengths of various pedicle screw designs are compared using tapped and untapped pilot holes. OBJECTIVE: The objective of this study is to compare the pullout strength of various pedicle screw designs. The designs are compared using tapped and untapped pilot holes. By using several different screw designs, it is possible to gain an understanding of whether there is a correlation between tapping a pilot hole and the ultimate pullout strength. SUMMARY OF BACKGROUND DATA: Most bone screws originally developed were intended to be installed in a pretapped pilot hole. This same technology has been carried over to the development of more modern bone screws for use in spinal fixation applications. Many pedicle screws in use today are still intended to be installed in a tapped hole. Preparing the vertebrae and tapping of a pilot hole involve additional trauma to the patient as well as increased operating time. METHODS: Pedicle screws from various manufacturers are installed in tapped and untapped pilot holes and then loaded to failure. A uniform synthetic material was used to provide a consistent test of each screw design by eliminating variability seen in bone. RESULTS: Tapping pilot holes did not increase the pullout strength of the screws tested in this study. It was observed during testing that tapping some of the holes degraded the material. This degradation led to pullout strengths that were lower than in the untapped case, and generally larger standard deviations. CONCLUSIONS: The pullout strength was not increased by tapping for the screws in this study. Screws placed in untapped holes generally had higher pullout strengths and lower standard deviations. The results of this study suggest that tapping does not increase pullout strength in bone with densities near 20 lb/ft3, which correlates with low density cancellous or osteoporotic bone. PMID- 17077723 TI - Comparative study of radiographic disc height changes using two different interbody devices for transforaminal lumbar interbody fusion: open box vs. fenestrated tube interbody cage. AB - STUDY DESIGN: Retrospective comparative study of the postoperative subsidence of two interbody devices following posterior or transforaminal lumbar interbody fusion (PLIF/TLIF) for degenerative spondylolisthesis of the lumbar spine. OBJECTIVE: To assess certain radiograph characteristics of PLIF/TLIF using two interbody fusion devices at L4-L5. SUMMARY OF BACKGROUND DATA: PLIF can achieve spinal stabilization with vertebral body support and direct neural decompression. Although various interbody devices have been used in PLIF procedures, no radiographic studies have compared the load-bearing capabilities of open box and fenestrated tube interbody cages. METHODS: Seventy-five patients who underwent one-level PLIF in the L4-L5 [corrected] segment for degenerative spondylolisthesis were retrospectively reviewed with a minimum 2-year follow-up. Fenestrated tube (Group FT: n = 30 [corrected]) or open box (Group OB: n = 45 [corrected]) cages were used for the PLIF procedure. The following radiographic parameters were evaluated to compare the load-bearing capabilities: disc space height (DH); percent increase and decrease of disc height (% IDH and % DDH, respectively); and percent coverage of the cage on the endplate (% CC). RESULTS: There were no significant differences in the baseline data, including age, segmental instability and osteoporotic status, between the two groups. Anterior %IDH and % CC were significantly higher in Group OB than in Group FT (% IDH: 69.4% vs. 57.3%; % OC: 24.5% vs. 12.9%), and anterior and posterior % DDH were significantly higher in Group FT than in Group OB (anterior: -2.9% vs. -.1%; posterior: -6.6% vs. -.3%). Although the restored DH gradually reduced over time in both groups, significant reduction to the preoperative level only occurred in Group FT. CONCLUSIONS: The load-bearing capabilities of the open box cage are superior to those of the fenestrated tube cage. Since there were no significant differences between the baseline status of the two groups, the larger cross sectional area and stable framework design of the open box cage appears to bring about a greater load-bearing capability. Therefore, the open box cage seems to be biomechanically more advantageous as an interbody device for PLIF than the fenestrated tube cage. PMID- 17077724 TI - Changes in the flexion relaxation response following an exercise intervention. AB - STUDY DESIGN: Pre and post 12-week training study using surface electromyography to measure the flexion relaxation response. OBJECTIVE: To evaluate whether the active or passive phases of the flexion relaxation measurement changes following an exercise intervention in patients with low back pain. SUMMARY OF BACKGROUND DATA: Impaired neuromuscular activation is an area of specific interest in patients with chronic nonspecific low back pain (LBP). The flexion relaxation phenomenon is commonly measured in LBP patients; however, there is insufficient evidence about the changes in this measure following an intervention. METHODS: Fifteen subjects with chronic LBP (7 females, 8 males) performed a 12-week training intervention. The main outcome measures were the Oswestry disability index, visual analog scale, and flexion relaxation response analyzed by the raw electromyograph (EMG) signal, the relative EMG signal, and the flexion relaxation ratio. RESULTS: Disability and pain scores improved significantly after the 12 week intervention. There were no changes in the active components of the flexion relaxation measurement but an approximate 67% decrease in the amount of activity measured during the relaxation phase at full trunk flexion. CONCLUSION: The data suggest that afferent feedback changes may be explaining why there is improved electrical relaxation following an exercise intervention. PMID- 17077725 TI - Patient and staff dosimetry in vertebroplasty. AB - STUDY DESIGN: Eleven vertebroplasty operations were studied in terms of radiation dose. OBJECTIVE: Doses to patients and staff associated with vertebroplasty were measured. Occupational doses were compared with the annual dose limits, and the effectiveness of the used radiation protection means was estimated. Patient dose was estimated by means of both surface and effective dose, and the radiation induced risk was evaluated. SUMMARY OF BACKGROUND DATA: Vertebroplasty is a recent minimally invasive technique for the restoration of vertebral body fractures. It involves fluoroscopic exposure, and so, it demands dose measurements for both patient and staff exposed to radiation. METHODS: Thermoluminescent dosimeters (TLDs) were placed on the medical personnel and the effective dose was derived. Slow films were placed to patients' skin to measure entrance surface dose. Furthermore, a Rando phantom loaded with TLDs was irradiated under conditions simulating vertebroplasty, in order to estimate effective dose to the patient. RESULTS: Mean fluoroscopy time was 27.7 minutes. Patient's mean skin dose was 688 mGy, while effective dose was calculated to be 34.45 mGy. It was estimated that the primary operator can perform about 150 vertebroplasty operations annually without exceeding the annual dose constraints, whereas occupational dose can be reduced by 76% using mobile shielding. CONCLUSIONS: Measures have to be taken to reduce patient's skin dose, which, in extreme cases, may be close to deterministic effects threshold. The highest dose rates, recorded during the procedure, were found for primary operator's hands and chest when no shielding was used. PMID- 17077727 TI - Endovascular embolization of iatrogenic vertebral artery injury during anterior cervical spine surgery: report of two cases and review of the literature. AB - STUDY DESIGN: Case description. OBJECTIVES: To report 2 cases of vertebral artery injury (VAI) during anterior cervical decompression surgery and review the pertinent medical literature. SUMMARY OF BACKGROUND DATA: The incidence of VAI during an anterior surgical approach to the cervical spine is rare but potentially lethal. The spine surgeon should be aware of the detailed surgical anatomy and be prepared to manage an inadvertent injury to the vertebral artery. METHODS: In the first case presented, infection was the cause of VAI. During aggressive irrigation and pus drainage, massive bleeding was encountered. For the second case, the vertebral artery was injured during decompression of cervical spondylosis while drilling the neural foramen. Intraoperative direct packing with hemostatic agents provided effective control of hemorrhage. RESULTS: Ten days after surgery, sudden neck swelling and mental deterioration occurred because of rebleeding from a pseudoaneurysm in the first case. In the second case, a pseudoaneurysm was detected by angiography, obtained 4 days after surgery. Both pseudoaneurysms were successfully occluded by an endovascular technique without any neurologic sequelae. CONCLUSIONS: In case of bleeding control by hemostatic packing, there remains a risk of delayed hemorrhage from pseudoaneurysm. Postoperative vertebral angiography is helpful to avoid life-threatening complications. Endovascular treatment can be a good alternative in the treatment of VAI. PMID- 17077728 TI - Benign intraspinal osteoblastoma stemming from C7 lamina in cervicothoracic junction: a case report. AB - STUDY DESIGN: A case report of a rare benign osteoblastoma successfully treated with total en bloc excision. OBJECTIVES: To describe the clinical and radiographic features of an adolescent female patient with intraspinal osteoblastoma in cervicothoracic junction and the effect of surgical treatment. SUMMARY OF BACKGROUND DATA: A rare case of massive benign osteoblastoma stemming from left side lamina of C7 and extension to spinal canal from C6 to T1 resulting in spinal cord compressed severely is reported. And the excellent results achieved from surgical intervention are presented. METHODS: A 13-year-old girl complained of gradually increasing pain of her lower part of the neck and left shoulder radiating to the left upper extremity, and the neurologic deficits were found in physical examination. A massive neoplasm located in intraspinal cervicothoracic junction and the spinal cord compressed severely was shown in radiographs, MRI, and CT. The neoplasm was removed totally with instrumentation performed in surgical intervention. She was followed up for 2.5 years; clinical symptoms and plain radiograph and CT films were investigated. RESULTS: The specimen of the removed mass was proven osteoblastoma pathologically. Her symptoms were relieved and the neurologic deficits recovered completely. Recurrence of the tumor was not seen at the final follow-up. CONCLUSIONS: A case of massive intraspinal osteoblastoma stemming from C7 lamina was successfully treated by excised grossly with instrumentation for reconstruction of stability. PMID- 17077730 TI - The effect of infliximab, a monoclonal antibody against TNF-alpha, on disc herniation resorption: a randomized controlled study. AB - STUDY DESIGN: Randomized, controlled study. OBJECTIVE: To evaluate the effect of infliximab on herniated nucleus pulposus (HNP) resorption. SUMMARY OF BACKGROUND DATA: Although the effects of tumor necrosis factor alpha (TNF-alpha) on HNP resorption are not fully understood, TNF-alpha appears to be an essential mediator in HNP resorption. METHODS: As part of a substudy of the FIRST II study, magnetic resonance images (MRIs) were obtained from 21 patients who were candidates for discectomy at weeks 0, 2, 12, and 26 after receiving a single infusion of either 5 mg/kg infliximab (11 patients) or placebo (10 patients). The volume (mm3) of HNP, thickness (mm) and extent (%) of rim enhancement, and presence of nerve root edema were assessed. RESULTS: HNP volume decreased significantly from baseline to 6 months in both treatment groups (P < 0.01), with no difference noted between the infliximab and placebo groups. By week 2, rim enhancement thickness increased significantly in the infliximab group compared with the placebo group (P = 0.003). Two patients in each group required back surgery before the 6-month assessment. CONCLUSIONS: Infliximab did not appear to interfere with disc herniation resorption over a 6-month period. PMID- 17077731 TI - Is fusion necessary for surgically treated burst fractures of the thoracolumbar and lumbar spine?: a prospective, randomized study. AB - STUDY DESIGN: A prospective clinical trial was conducted. OBJECTIVES: To compare the results of fusion versus nonfusion for surgically treated burst fractures of the thoracolumbar and lumbar spine. SUMMARY OF BACKGROUND DATA: The operative results of surgically treated burst fractures with short segmental fixation have been well documented. There is no report comparing the results of fusion and nonfusion. METHODS: Fifty-eight patients were included in this study, with the inclusion criteria as follows: neurologically intact spine with a kyphotic angle > or = 20 degrees, decreased vertebral body height > or = 50% or a canal compromise > or = 50%, incomplete neurologic deficit with a canal compromise 50%, complete neurologic deficit, and multilevel spinal injury or multiple traumas. All patients were randomly assigned to fusion or nonfusion groups, and operative treatment with posterior reduction and instrumentation was carried out. Posterior fusion with autogenous bone graft was performed for the fusion group (n = 30), and no fusion procedure was done for the nonfusion group (n = 28). The average follow-up period was 41 months (range, 24-71 months). RESULTS: The average loss of kyphotic angle was not statistically significant between these 2 groups. The radiographic parameters were statistically significantly better in the nonfusion group, including angular change in the flexion-extension lateral view (4.8 degrees vs. 1.0 degrees), lost correction of decreased vertebral body height (3.6% vs. 8.3%), intraoperative estimated blood loss (303 mL vs. 572 mL), and operative time (162 minutes vs. 224 minutes). The scores on the low back outcome scale were not statistically significant for these 2 groups. CONCLUSIONS: The short-term results of short segmental fixation without fusion for surgically treated burst fractures of the thoracolumbar spine were satisfactory. The advantages of instrumentation without fusion are the elimination of donor site complications, saving more motion segments, and reducing blood loss and operative time. PMID- 17077733 TI - The reciprocal relationship between thoracic and spinal deformity and its effect on pulmonary function in a rabbit model: a pilot study. AB - STUDY DESIGN: Investigate the association between growth of the spine and thorax under conditions that create symmetric or asymmetric growth disturbances of the spine or thorax in a growing rabbit. OBJECTIVE: Prove growth of the spine and growth of the thorax are directly related. SUMMARY OF BACKGROUND DATA: Understanding the association between growth of the thorax and growth of the spine may explain the occurrence of thoracic insufficiency in patients with congenital scoliosis. METHODS: Symmetric or asymmetric growth disturbances of the spine or thorax were established in 5-week-old rabbits. During growth of the rabbits, structural changes in the spine, thorax, and lung volume were assessed using serial CT scans. Measures of pulmonary function, spine, and thoracic deformity were related to one another across groups. RESULTS: The mean Cobb angle and distortion of the thoracic cage were significantly greater for the unilateral tethered rib group. There were no significant differences in total lung volume among the experimental groups. However, the left/right lung volume ratio was significantly different for the unilateral tethered rib group compared with the control group. CONCLUSIONS: Unilateral deformity of the spine or thorax induces both a scoliosis and thoracic cage deformity with asymmetric lung volumes. PMID- 17077734 TI - An in vitro organ culturing system for intervertebral disc explants with vertebral endplates: a feasibility study with ovine caudal discs. AB - STUDY DESIGN: Whole ovine caudal intervertebral discs with vertebral endplates were cultured under uniaxial diurnal loading for 7 days. OBJECTIVES: To establish and characterize an organ culture system for intervertebral discs, in which disc cells may be "maintained" in their native three-dimensional environment under load. SUMMARY OF BACKGROUND DATA: In vitro culturing of entire discs with preserved biologic and structural integrity would be a useful model to study the effects of nutrition and mechanical loading. METHODS: To maintain endplate permeability, sheep were systemically anticoagulated before death and their caudal vasculature was evacuated with saline postmortem. The first 4 caudal discs were explanted with their adjacent endplates and cultured in bioreactors under uniaxial diurnal loading (0.2 MPa for 8 hours and 0.8 MPa for 16 hours) for 4 or 7 days. Solute transport into the center of the disc was measured after 4 days of culture using a low molecular weight fluorescent marker. Cell viability, glycosaminoglycan synthesis rate, and gene expression profile were measured after 7 days of culture and compared with fresh tissue. RESULTS: Fluorescent images showed that solutes could diffuse into the disc under both static and diurnal loading, but penetration through the endplate increased with diurnal loading. Cell viability and glycosaminoglycan synthesis rates remained unchanged after 7 days of culture. Expression of catabolic genes was significantly up-regulated, whereas anabolic genes tended to be down-regulated after 7 days. CONCLUSIONS: With this novel preparation and culturing technique, endplate permeability could be maintained, which allowed culturing of intact disc explants with endplates for up to 7 days. PMID- 17077735 TI - Hematologic abnormalities within the first week after acute isolated traumatic cervical spinal cord injury: a case-control cohort study. AB - STUDY DESIGN: Case-control cohort study. OBJECTIVE: To evaluate 1) the hematologic abnormalities within the first week following isolated acute cervical spine trauma or spinal cord injury (SCI); and 2) the influence of age, sex, and severity of SCI on these hematologic abnormalities. SUMMARY OF BACKGROUND DATA: Given that autonomic nervous system has a critical role in the regulation of the hematopoietic system, we sought to evaluate the potential association between hematologic abnormalities within the first week posttrauma and the severity of SCI. METHODS: All consecutive individuals with isolated acute cervical spine trauma admitted to our institution from 1998 to 2000 were reviewed. Exclusion criteria included preexisting medical comorbidities and polytrauma. The study population was divided into a SCI group and a control group (individuals with spine trauma without neurologic impairment). The SCI group was subdivided into patients with motor complete SCI (American Spinal Injury Association [ASIA] Grades A/B) and individuals with motor incomplete SCI (ASIA Grades C/D). RESULTS: There were 21 SCI individuals (15 male, 6 female; ages 17-83 years; mean, 57 years) and 11 controls (6 male, 5 female; ages 18-75 years; mean, 41 years). When controlled for age, SCI individuals showed a significantly higher frequency of reduced hemoglobin concentration (RHC), leukocytosis, lymphopenia, and thrombocytopenia than controls within the first week posttrauma. Blood hemoglobin concentration, lymphocyte, and platelet counts in the SCI group were significant lower than the controls. The SCI group showed a significant higher leukocyte count than the controls. The degree of RHC and lymphopenia was significantly correlated with the severity of SCI. CONCLUSION: Our results indicate that patients with isolated cervical SCI have significantly greater frequency of RHC, leukocytosis, lymphopenia, and thrombocytopenia than controls during the first week posttrauma. The degree of RHC and lymphopenia was significantly associated with the severity of SCI. While the mechanisms underlying these clinically important hematologic consequences of SCI remain undetermined, evidence from the literature suggests that acute autonomic denervation of the hematopoietic system could play a key role. PMID- 17077736 TI - Segmental motor paralysis after cervical laminoplasty: a prospective study. AB - STUDY DESIGN: A prospective study. OBJECTIVE: To examine the prevalence and clinical manifestations of segmental motor paralysis after cervical laminoplasty and to investigate the presence of intramedullary high-signal intensity area (HIA) on postoperative T2-weighted magnetic resonance imaging (MRI). SUMMARY OF BACKGROUND DATA: Almost all previous studies had been retrospectively conducted, and MRIs of all subjects, including a control population, have never been investigated. METHODS: In the 79 patients of the prospective group, the sole examiner evaluated muscle strengths daily for perioperative 3 weeks and MRI scans were performed before and after surgery. Medical records of the 79 consecutive patients before this study were also investigated (control group). RESULTS: Ten patients from the prospective group developed segmental motor paralysis (proximal in 5, distal in 2, and diffuse in 3), whereas paralysis occurred in only 4 patients from the control group (proximal in 3 and diffuse in 1). On postoperative MRI, a linear HIA corresponded to the paralyzed segment more frequently than a focal or no HIA. CONCLUSION: In the prospective cases, distal or diffuse paralysis was found more frequently than in the retrospective control group. Linear HIAs were significantly more likely to present in the paralyzed segments. PMID- 17077737 TI - Microendoscopic discectomy for lumbar disc herniation: surgical technique and outcome in 873 consecutive cases. AB - STUDY DESIGN: A retrospective review involving 873 consecutive cases of lumbar disc herniation treated by microendoscopic discectomy (MED) was conducted and a mean 28-month follow-up was performed. OBJECTIVE: The purpose of this study was to describe the MED technique for lumbar disc herniation and report long-time outcome and complications. SUMMARY OF BACKGROUND DATA: The conception of MED was introduced in 1997. Long-time outcome has not been described. METHOD: A total of 873 consecutive patients with lumbar disc herniation were treated with the METRx system. Oswestry Disability Index (ODI) was used to quantify pain relief. The degree of pain and disability was also measured by visual analog scale (VAS) and modified MacNab criteria. A control group of 358 patients treated with standard open discectomy was used for comparison. RESULTS: There was significant improvement in the mean preoperative and postoperative VAS and ODI score for the MED and open groups, and there was no statistical difference of the pain improvement between the two groups. For the MED group, average length of hospital stay was 4.8 days; mean time to return to work or normal activities was 15 days; average operative blood loss per level was 44 mL. These were significantly less than open group. CONCLUSIONS: MED is an effective microendoscopic system with fine long-term outcome in treating lumbar disc herniation. The endoscopic approach allows smaller incisions and less tissue trauma, compared with standard open microdiscectomy. Strict adherence to well-defined preoperative selection criteria could ensure optimal postoperative outcome. PMID- 17077738 TI - Nontuberculous pyogenic spinal infection in adults: a 12-year experience from a tertiary referral center. AB - STUDY DESIGN: We performed a retrospective review of 48 cases of pyogenic spinal infection presenting over a 12-year period to the National Spinal Injuries Unit (NSIU) of the Republic of Ireland. The NSIU is the tertiary referral center for all adult spinal injuries and diseases of the spine warranting surgical intervention in the Republic of Ireland. OBJECTIVES: The objective of this study was to analyze the presentation, etiology, management, and outcome of nontuberculous pyogenic spinal infection in adults. SUMMARY OF BACKGROUND DATA: Pyogenic spinal infection encompasses a broad range of clinical entities, including spondylodiscitis, septic discitis, vertebral osteomyelitis, and epidural abscess. Management of pyogenic spinal infection can involve conservative methods and surgical intervention. METHODS: The medical records, radiologic imaging, and bacteriology results of 48 patients with pyogenic vertebral osteomyelitis from 1992 through 2004 were reviewed. The Hospital Inpatient Enquiry (HIPE) System and the NSIU Database were used to identify our study cohort. RESULTS: The average age of presentation was 59 years with an even distribution between males and females. Most patients (21 of 48) were symptomatic for between 2 and 6 weeks before presenting to hospital. The most frequently isolated pathogen was Staphylococcus aureus, in 23 of 48 cases (48%); 35 of 48 cases (73%) were managed by conservative measures alone, including antibiotic therapy and spinal bracing. However, in 13 of 48 cases (27%), surgical intervention was required because of neurologic compromise or mechanical instability. CONCLUSIONS: In the majority of cases, conservative management of pyogenic spinal infection with antibiotic therapy and spinal bracing is very successful. However, in a minority of cases, surgical intervention is warranted and referral to a specialist center is appropriate. PMID- 17077739 TI - Magnetic resonance imaging of the lumbar spine: findings in female subjects from administrative and nursing professions. AB - STUDY DESIGN: Cross-sectional magnetic resonance imaging (MRI) study. OBJECTIVE: To find out whether different patterns of lumbar disc degeneration related to different physical work types (static vs. dynamic) can be found. SUMMARY OF BACKGROUND DATA: The association of lumbar disc degeneration with physical workload as well as the association of disc degeneration with low back pain (LBP) has been debated controversially. Nevertheless, many of the mainly invasive treatment concepts are based on disc degeneration models. METHODS: An MRI study was performed in female subjects aged 45 to 62 years with persistent LBP and in age-matched controls. Subjects (n = 109) were selected from nursing and administrative professions. A questionnaire was used to assess the subjects' exposure to physical workplace factors. Disc degeneration, disc herniation, nerve root compromise, high intensity zones, endplate changes, and facet joint osteoarthritis were evaluated based on sagittal T1- and T2-weighted as well as axial T2-weighted images. RESULTS: The two occupational groups could clearly be distinguished by the physical workplace factors: administrative work was associated with predominantly sitting postures whereas nursing work was associated with frequent walking, pushing, pulling, and lifting. Except for endplate (Modic) changes at L5-S1, MRI findings did not differ between the four groups, namely, nurses and secretaries with and without LBP. When analyzing the MRI findings within the two vocational groups, nerve root compromise and endplate changes in the lower lumbar spine were found to be significant (P < 0.05) risk factors for LBP. Disc degeneration, disc herniation, nerve root compromise, and facet joint osteoarthritis were present in more than 50% of the intervertebral spaces studied. CONCLUSIONS: These findings give evidence that in subjects performing nonheavy work, patterns of lumbar disc degeneration are not associated with the job type and characteristic physical loadings. PMID- 17077740 TI - United States' trends and regional variations in lumbar spine surgery: 1992-2003. AB - STUDY DESIGN: Repeated cross-sectional analysis using national Medicare data from the Dartmouth Atlas Project. OBJECTIVE: To describe recent trends and geographic variation in population-based rates of lumbar fusion spine surgery. SUMMARY OF BACKGROUND DATA: Lumbar fusion rates have increased dramatically during the 1980s and even more so in the 1990s. The most rapid increase appeared to follow the approval of a new surgical implant device. METHODS: Medicare claims and enrollment data were used to calculate age, sex, and race-adjusted rates of lumbar laminectomy/discectomy and lumbar fusion for fee-for-service Medicare beneficiaries over age 65 in each of the 306 US Hospital Referral Regions between 1992 and 2003. RESULTS: Lumbar fusion rates have increased steadily since 1992 (0.3 per 1000 enrollees in 1992 to 1.1 per 1000 enrollees in 2003). Regional rates of lumbar discectomy, laminectomy, and fusion in 1992-1993 were highly correlated to rates of discectomy, laminectomy (R2 = 0.44), and fusion (R2 = 0.28) in 2002-2003. There was a nearly 8-fold variation in regional rates of lumbar discectomy and laminectomy in 2002 and 2003. In the case of lumbar fusion, there was nearly a 20-fold range in rates among Medicare enrollees in 2002 and 2003. This represents the largest coefficient of variation seen with any surgical procedure. Medicare spending for inpatient back surgery more than doubled over the decade. Spending for lumbar fusion increased more than 500%, from 75 million dollars to 482 million dollars. In 1992, lumbar fusion represented 14% of total spending for back surgery; by 2003, lumbar fusion accounted for 47% of spending. CONCLUSIONS: The rate of specific procedures within a region or "surgical signature" is remarkably stable over time. However, there has been a marked increase in rates of fusion, and a coincident shift and increase in cost. Rates of back surgery were not correlated with the per-capita supply of orthopedic and neurosurgeons. PMID- 17077741 TI - Lumbar fusion outcomes in Washington State workers' compensation. AB - STUDY DESIGN: Retrospective population-based cohort study. OBJECTIVE: To evaluate the influence of lumbar intervertebral fusion devices on clinical and disability outcomes among Washington State compensated workers with chronic back pain. SUMMARY OF BACKGROUND DATA: The efficacy of lumbar fusion for chronic low back pain remains controversial. Recent randomized controlled trials have shown results of fusion to be equivalent to those of structured exercise and cognitive intervention. Lumbar fusion rates, however, continue to increase nationally, fueled, in part, by introduction of new fusion devices, including intervertebral cages in 1996. It is not known whether these newer devices have improved outcomes, and particularly in compensated workers. METHODS: In this population based retrospective cohort study, we identified Washington State injured workers who underwent lumbar fusion between 1994 and 2001 from Washington State Workers' Compensation system administrative database. All the data for this study were obtained from either the claims or medical bill payment databases. Multiple logistic regression analyses were used to examine the association between the surgical technique (interbody cages and/or instrumentation) and the risk of disability, reoperation, and complications following lumbar fusion. RESULTS: Among the 1,950 eligible subjects, fusions with cages increased from 3.6% in 1996 to 58.1% in 2001. Overall disability rate at 2 years after fusion was 63.9%, reoperation rate 22.1%, and rate for other complications 11.8%. Use of cages or instrumentation was associated with increased complication risk compared with bone-only fusions without improving disability or reoperation rates. Legal, work related, and psychologic factors predicted worse disability. Discography and multilevel fusions predicted greater reoperation risk. Degenerative disc disease and concurrent decompression procedures predicted lower reoperation risk. CONCLUSIONS: Use of intervertebral fusion devices rose rapidly after their introduction in 1996. This increased use was associated with an increased complication risk without improving disability or reoperation rates. PMID- 17077742 TI - Back pain prevalence and visit rates: estimates from U.S. national surveys, 2002. AB - STUDY DESIGN: Review and analysis of data from two U.S. national surveys in 2002. OBJECTIVES: To examine the prevalence of back pain and physician visits for back pain in the United States. SUMMARY OF BACKGROUND DATA: National data on the prevalence of back pain become available only intermittently. METHODS: We summarized published data from the 2002 National Health Interview Survey (NHIS) on the prevalence of back pain and compared it with earlier surveys. We also analyzed the 2002 National Ambulatory Medical Care Survey (NAMCS) to determine physician visit rates for back pain. RESULTS: In the 2002 NHIS, there were 31,044 adult respondents. Low back pain lasting at least a whole day in the past 3 months was reported by 26.4% of respondents, and neck pain was reported by 13.8%. Among racial groups, American Indians and Alaska Natives had the highest prevalence of low back pain, and Asian Americans had the lowest. Prevalence generally declined with greater levels of education and increasing income. Prevalence estimates were consistent with those from previous surveys, although methodologic differences limited comparisons. NAMCS data suggested that the proportion of all physician visits attributable to low back pain (2.3% in 2002) has changed little since the early 1990s. CONCLUSIONS: About one fourth of U.S. adults report low back pain in the past 3 months; the proportion of physician visits attributed to back pain has changed little in the past decade. PMID- 17077744 TI - Pain drawing scoring is not improved by inclusion of patient-reported pain sensation. AB - STUDY DESIGN: This is a retrospective study of 250 patients who describe low back pain with pain drawings. A computer application using artificial neural networks was designed to analyze pain drawings and evaluate the contribution of pain sensation to drawing classification. OBJECTIVE: The primary goal of this study was to assess the contribution of patient recorded pain sensation marks in classifying pain drawings into one of five broadly defined categories. The hypothesis was that including pain sensation would improve classification. SUMMARY OF BACKGROUND DATA: With no perfect diagnostic test for patients with low back pain, many approaches have been proposed and are used. One common diagnostic tool is the pain drawing. Several quantitative methods have been proposed to score the drawings. Some methods use pain sensation in the scoring; however, the contribution of pain sensation has not been defined. METHODS: A custom computer application classified the pain drawing. Data consisted of 250 pain drawings from patients with low back pain. RESULTS: Patient recorded pain sensation is not necessary in computer-based scoring of pain drawings. CONCLUSION: Patient reported pain sensation does not improve classification when quantitatively scoring pain drawings. PMID- 17077743 TI - Prevalence and physical determinants of low back pain in a rural Chinese population. AB - STUDY DESIGN: This was a community-based cross-sectional study that is part of an ongoing study of genetics and osteoporosis. OBJECTIVES: To estimate the 1-year self-reported prevalence of low back pain (LBP); and to assess the association between physical exposures (occupation, physical stress, and vibration) and LBP in a rural Chinese population. This study also explored how these associations change when individuals with LBP report additional pains in other parts of the body. SUMMARY OF BACKGROUND DATA: Little is known about the prevalence and determinants of LBP in rural developing populations, even though approximately half of the world's population, mostly from the developing world, is engaged in agriculture, a known strenuous activity. METHODS: Families with at least three participating siblings residing in the surrounding rural region of Anqing city, Anhui Province, China, were invited to participate in the study through public announcements and direct invitations by village physicians. A questionnaire including demographics, physical exposures, and musculoskeletal pain was administered to 13,965 men and women (age, 25-64 years). Generalized estimating equations were used to estimate LBP prevalence and examine associations between LBP and physical exposures. RESULTS: The 1-year prevalence of LBP was 64%. Twenty five percent reported no pain at any body site. Women had higher prevalence of LBP than men across all age groups. Being a farmer, reporting moderate or heavy physical stress, and having had former or current exposures to vibration were positively associated with LBP (P < 0.05). Physical exposures were also associated with cases of LBP combined with other musculoskeletal pains, and generally, the more pains individuals reported, the more likely it was that they were farmers and were exposed to vibration. CONCLUSION: A high prevalence of LBP and LBP with additional musculoskeletal pain existed in this rural Chinese sample. We found evidence of a link between physical exposures and LBP, and LBP with additional musculoskeletal pain. Further understanding of the characteristics and risk factors of LBP in rural developing areas is needed. PMID- 17077746 TI - Polish cultural adaptation of the Roland-Morris Questionnaire for evaluation of quality of life in patients with low back pain. AB - STUDY DESIGN: Cultural adaptation and cross-sectional psychometric testing. OBJECTIVES: The aim of this study was to validate the Polish translation of the Roland-Morris Questionnaire (RMQ), which evaluates the functional level of activity of patients with discogenic low back pain with radicular radiation, caused by intervertebral herniation, according to category A1 in the Kirkaldy Willis classification and Type QTF6 in classification Quebec Task Force. SUMMARY OF BACKGROUND DATA: The main concern of patients with low back pain is the functional limitation that the symptoms cause. It is important to find a validation tool by means of which their functional level can be assessed. The RMQ was compared with MOS Short Form-36 Health Survey Questionnaire (SF-36), to pain scale, to depression scale, to functional scale that measures activities of daily living, and to measuring forward and side bending of the spine. METHODS: Independently, two professionals made a translation and then two native speakers made a translation-back of the RMQ. Then 104 patients with lumbar disc herniation completed the RMQ, SF-36, VAS and FIR (ADL scale). The study group comprised 39 women and 65 men with an average age of 44 years. RESULTS: The significant correlation between the RMQ and other questionnaires was found. RMQ and SF-36 Physical, r = 0.47, P < 0.00001, RMQ and Pain, r = 0.462, P < 0.00001. Cronbach's alpha showed a value of 0.88 (scale 0-1). CONCLUSIONS: The correlation between severity of disease and quality of life, between bending of spine and quality of life, between depression and quality of life, and between activities of daily living and quality of life has been found. According to this study, the RMQ can be used as a valid tool in the assessment of the functional level of patients with lumbar pain and previous lumbar herniation. The RMQ is fast and easy to complete; it is valid, reliable, and sensitive. We recommend the questionnaire to be used in research of quality of life in low back pain. PMID- 17077747 TI - Balloon kyphoplasty and vertebroplasty for vertebral compression fractures: a comparative systematic review of efficacy and safety. AB - STUDY DESIGN: Systematic review and meta-regression. OBJECTIVES: To compare the efficacy and safety of balloon kyphoplasty and vertebroplasty for the treatment of vertebral compression fractures, and to examine the prognostic factors that predict outcome. SUMMARY OF BACKGROUND DATA: A previous systematic review of vertebroplasty by Levine et al in 2000 identified seven case series studies and no controlled studies. METHODS: A number of electronic databases were searched through March 1, 2004. Citation searches of included studies were undertaken and contact was made with experts in the field. No language restrictions were applied. All controlled and uncontrolled studies were included with the exception of case reports. Prognostic factors responsible for pain relief and cement leakage were examined using meta-regression. RESULTS: The following studies were included: balloon kyphoplasty (three nonrandomized comparative studies against conventional medical therapy and 13 case series), vertebroplasty (one nonrandomized comparative study against conventional medical care and 57 cases series), balloon kyphoplasty versus vertebroplasty (one nonrandomized comparative study). The majority of studies were undertaken in older women with osteoporotic vertebral compression fractures with long-term pain that was refractory to medical treatment. At this time, there is no good quality direct comparative evidence of balloon kyphoplasty versus vertebroplasty. From indirect comparison of case series evidence, the procedures appear to provide similar gains in pain relief while for balloon kyphoplasty there is better documentation of gains in patient functionality and quality of life. The level of cement leakage and number of reported adverse events (pulmonary emboli and neurologic injury) in balloon kyphoplasty was significantly lower than for vertebroplasty. These findings were confirmed by meta-regression analysis. CONCLUSIONS: There is Level III evidence to support balloon kyphoplasty and vertebroplasty as effective therapies in the management of patients with symptomatic osteoporotic vertebral compression fractures refractory to conventional medical therapy. Although there was a good ratio of benefit to harm for both procedures, balloon kyphoplasty appears to offer the better adverse event profile. These conclusions need to be updated on the basis of the findings of ongoing randomized controlled trials. PMID- 17077749 TI - Re: Rampersaud YR, Moro ER, Neary MA, et al. Intraoperative adverse events and related postoperative complications in spine surgery: implications for enhancing patient safety founded on evidence-based protocols. Spine 2006;31:1503-10. PMID- 17077750 TI - Cortical activations, psychiatric symptoms, and climacteric women. PMID- 17077751 TI - Medial frontal cortex perfusion abnormalities as evaluated by positron emission tomography in women with climacteric symptoms. AB - OBJECTIVE: To identify functional changes in the brains of women with climacteric symptoms. Images of regional cerebral blood flow (rCBF) were compared statistically between women with and women without symptoms to identify changes in rCBF. Results may provide a better understanding of the neural basis of the symptoms, which are divided into three symptom clusters: vasomotor, psychological, and somatic. DESIGN: The study participants consisted of 12 women with moderate to severe climacteric symptoms (age 47.5 +/- 5.9 years, mean +/- SD) and 7 women with no symptoms (control group; age 49.6 +/- 4.2 years, mean +/- SD). The study participants were patients at a menopause clinic, and the latter were healthy volunteer nurses and hospital staff. Climacteric symptoms were evaluated by an assessment of the severity of 17 symptoms immediately before positron emission tomography examination of rCBF. The symptoms had been used previously to generate the Kupperman Kohnenki Shogai Index, a modified Kupperman Menopausal Index adapted to Japanese women. rCBF was measured by positron emission tomography with the CO2 dynamic inhalation method. RESULTS: Reductions in relative rCBF in the patient group were observed in the bilateral rectal gyrus and in the left subcallosal gyrus on a voxel-by-voxel basis as compared with the control group. CONCLUSIONS: The present study revealed reductions in relative rCBF of the prefrontal cortex of Japanese women with moderate to severe climacteric symptoms. This area is close to that previously addressed in studies of familial bipolar depression and familial unipolar depression, although our participants did not satisfy criteria for depression. This reduction of rCBF may be related to the three climacteric symptom clusters, but further studies are needed for evaluation of its significance. Our results should stimulate investigations into the positron emission tomography rCBF change of these women as to the integration of multiple entities in climacteric symptoms. PMID- 17077752 TI - Implementing the NICE pressure ulcer guideline. AB - This article explores the practical implementation of the NICE (2005) guideline on pressure ulcer prevention by those that the guideline was created for, including: service managers, commissioners, clinical governance and education leads, patients/carers and both the specialist and non-specialist. PMID- 17077753 TI - Alternative ways of managing access to wound products. PMID- 17077754 TI - Easing compression hosiery application: ActiGlide. PMID- 17077755 TI - Evidence, prescribing and the Drug Tariff. PMID- 17077756 TI - We listen but do we hear? The importance of patient stories. AB - Patients' perspectives on their illness and care are increasingly recognized as the focus for the organization of health care, yet patient stories are rarely examined on their own. Mostly, they are dissected for data, and so the impact is lost. This article presents a number of story tellers, all patients with leg wounds attending a social clinic. Reading their stories helps us understand the experience of being a patient and living with difficult and painful leg wounds. They also show how nurses can play a critical role in optimizing the power of the story in the patient's journey towards physical and psychological healing. PMID- 17077758 TI - Risks of long-term catheterization in the community--and managing them. AB - Catheter-associated urinary tract infections continue to be a major source of morbidity in hospital and the community. In part, this may be the result of ritualistic practices, compounded by poor communication on discharge from hospital. This article highlights many of the issues that lead to continued high rates of CAUTI, and challenges health professionals and health-care providers to use all available guidance and tools to reduce the incidence of CAUTI across the NHS. PMID- 17077759 TI - Challenges and strategies: weight management in type 2 diabetes. AB - This article examines the link between obesity and type 2 diabetes and discusses some of the challenges health-care professionals face in helping people with diabetes achieve and maintain a healthy weight. Increasing numbers of people in both developed and developing countries are being classified as overweight or obese, which is resulting in growing numbers of people being diagnosed with type 2 diabetes. Worryingly this includes large numbers of children developing type 2 diabetes as the younger age groups are becoming overweight or obese. The health care practitioner's role is explored and strategies to promote and achieve a healthy lifestyle for people with type 2 diabetes are offered. Such strategies are linked to obesity management through, diet, exercise and behavioural therapy. PMID- 17077760 TI - Validation and usefulness of the Tamil version of the GHQ-12 in the community. AB - The 12-item General Health Questionnaire (GHQ-12), increasingly used to screen for common mental disorders (CMD) in primary care, has been validated in different languages and cultures. However, the validity and usefulness of the Tamil version has not been established for use in the community. This article reports a study to validate the Tamil version of the GHQ-12 in a rural setting in southern India. The research found that 16.5% of people interviewed satisfied international criteria for CMD. However, the predictive value of the tool as a tool for screening was found to differ from that found in studies undertaken in a hospital setting. While screening instruments need to be validated in specific types of populations before being employed, such tests may be less useful in community surveys where the prevalence of common mental disorders is low. Strategies which artificially increase the prevalence of common mental disorders in the sample screened will improve the performance of the instrument. PMID- 17077761 TI - Understanding and tackling fear of crime among older people. AB - In this article, brought to you in association with Help the Aged, Alan Burnett examines the causes of fear of crime among older people, and proposes ways to reduce it. PMID- 17077762 TI - The Warrington workload tool: determining its use in one trust. AB - With recent advances in chronic disease management and with earlier discharge from the acute sector, it has become increasingly important for district nurses to monitor and evidence their increasing workload, to monitor demand and capacity. The district nursing service in the author's trust has employed an adaptation of the Warrington workload tool, over several years, to demonstrate and evidence their workload. However, many dispute the accuracy of such tools. This audit set out to discover how accurately the tool was being calculated. The findings have cast doubt on the tool's accurate completion and highlighted an ongoing training need to ensure improvement in its use. The tool is to be further supported with the inclusion of a caseload profiling tool to provide qualitative evidence of the service that the nurses now provide. It was also identified that the service needs evidence that their resource needs are met in response to workload tool data to ensure the tool continues to be used in a proactive manner. PMID- 17077763 TI - Compensation for psychiatric injury: evolution of a law of nervous shock. AB - District nurses will be well aware that if a person receives an injury because of someone else's negligence they are entitled to compensation for the harm that has been caused. However, where the injury is in the form of psychiatric harm the law has traditionally imposed rules that set out the conditions to be met before a successful claim for damages can be made. Even in today's enlightened society the law still does not always treat a psychiatric injury in the same way as a physical injury. In this article Richard Griffith outlines the development of the law in relation to psychiatric injury--historically called "nervous shock"--and considers the current approach to claims for damages where a person suffers psychiatric harm because of another's negligent act. PMID- 17077764 TI - Lessons from Stoke Mandeville. PMID- 17077765 TI - Breast cancer awareness: star, posh, hot and prime. PMID- 17077766 TI - Post-exposure prophylaxis:access and fairness. PMID- 17077767 TI - Risks and benefits of needle use in patients after axillary node surgery. AB - Lymphoedema may be triggered by any type of injection. This is not just an issue for the cured breast cancer patient but for any patient who has undergone under arm (axillary) lymph node removal to any degree for any type of cancer. Standard advice given to patients following axillary node removal is to avoid any injection or blood pressure measurement on the ipsilateral arm. The evidence base in this area is severely lacking. Patients have reported frustration that nurses are not informed of contraindications in carrying out such procedures on patients at risk of developing swelling. This article discusses the current evidence available on the subject of non-accidental skin puncture (NASP) relating to the patient at risk of lymphoedema and provides guidelines for any professionals conducting such procedures for patients with a history of cancer. The results from a small audit of the guidelines are cited and they reveal that out of 14 patients who underwent NASP procedures in the at risk arm, no patients reported swelling to that limb within a month of these procedures. PMID- 17077768 TI - Evidence for the prevention of venous thromboembolism. AB - Patients undergoing major orthopaedic surgery are at high risk of developing venous thromboembolism (VTE). VTE is preventable and venous prophylaxis consensus groups recommend that each patient is assessed for risk of VTE and then stratified into one of the three categories of risk. Rick stratification enables the choice of the most appropriate preventative interventions. This article examines a decision making framework for VTE prevention with particular focus on a validated risk assessment model (RAM) to facilitate risk stratification. The relevant literature is also scrutinised in terms of the best venous antithrombotic strategies, for patients undergoing major orthopaedic surgery, according to scientific evidence. PMID- 17077769 TI - Softform Premier Active mattress: a novel step-up/step-down approach. AB - In the UK it is estimated that as many as 412,000 patients who are already ill will develop pressure ulcers (Bennett et al, 2004), which are an unnecessary and expensive complication to treat (Hitch, 1995). One way to try and reduce these risks is by investing in suitable mattresses. The author's investigations into existing mattress stock in the years 1996-97 illustrated the inadequacy of the NHS standard mattress (Santy, 1995; Fox, 1997). Investigators (e.g. Rithalia, 1996) were recognizing the inadequacy of the pink marbled standard NHS foam mattress for pressure reduction (Medical Device Directorate, 1993; Dunford, 1994) which included reports necessary to help develop guidelines in pressure ulcer prevention and management (Coull, 2004), as well as make value-for-money recommendations about product purchases (Fletcher et al, 1994; Value For Money Update, 1994; Cullum et al, 1995). PMID- 17077770 TI - Care of patients with haemoglobin abnormalities: history and biology. AB - Haemoglobinopathies refer to a range of genetically inherited disorders of red blood cell haemoglobin and include sickle cell disorders and thalassaemias. They occur most commonly in populations whose ancestors come from Africa, Asia, Mediterranean Islands, and the Middle and Far East. Haemoglobin (Hb) abnormalities (or haemoglobinopathies) are caused by (i) abnormalities of the protein structure; (ii) imbalanced globin chain production owing to reduced rate of synthesis of normal a or b globin chains; or (iii) a combination of the two. This article will focus on the biological basis of sickle cell disorders and will discuss the history and pathology of the conditions. PMID- 17077771 TI - Pathophysiology, diagnosis and management of acute pancreatitis. AB - Pancreatitis is an inflammation of the pancreas that can be either acute or chronic in origin and is caused by a varied range of aetiologies. However, in both types of the disease alcohol use may be a major contributor. In acute pancreatitis the symptoms can range from mild, which resolve within a few days, to severe, which is associated with both multi-organ failure and a high mortality necessitating admission to a high dependency or intensive care unit. As the incidence of acute pancreatitis is increasing it is imperative that nurses are familiar with the causes, underlying pathophysiology, diagnostic tests and the potentially vast range of complications that can occur in acute pancreatitis. This article reviews the current medical and nursing management of this condition. PMID- 17077772 TI - Treatment decision following a fingertip injury. PMID- 17077773 TI - Differential diagnosis in advanced nursing practice. AB - Differential diagnosis has become a key element of advanced nursing roles that are incorporating history taking and physical assessment into their remit of professional responsibilities. The aim of this article is to review the relevant definitions of different types of diagnoses, to describe the process of formulating a differential diagnosis including using clinical reasoning skills, and to use a case study as an example of differential diagnosis in advanced clinical practice. PMID- 17077774 TI - Comparing no-touch and tympanic thermometer temperature recordings. AB - Temperature is a vital sign which can be measured using various types of clinical thermometers. Pulmonary artery temperature is considered the 'gold standard', but this measurement is not usually clinically practical. There is currently no consensus for optimal alternative site or equipment. This research compares 178 simultaneous measurements from 5 clinical areas, using two types of thermometers: tympanic and no-touch temporal. No-touch thermometers were all set to oral equivalent. Tympanic thermometers were adjusted to either oral (n=105) or core (n=73) equivalent. Maximum acceptable difference was identified as 1oC. Two data sets (oral/core; oral/oral) were analysed using Bland-Altman method on Excel programmes, comparing all thermometers and separating oral and core-equivalent tympanics. The two thermometers were found not to be equivalent. As a simple comparison between two thermometers, this research cannot identify which thermometer is more accurate. PMID- 17077775 TI - Reductionism and holism in coronary heart disease and cardiac nursing. AB - Reductionism and holism are two contrasting philosophies which provide scientific knowledge of disease processes, health dynamics and care interventions. While reductionism focuses on specific and perhaps narrow concepts, it enhances our in depth knowledge of key health issues. Holism focuses on understanding how all the significant factors affecting the particular health issue are involved, so a more informed decision can be made about health intervention. This article explores the contribution each makes to our understanding of coronary heart disease (CHD) and to the preparation of nurses working in cardiac nursing. It proposes that pre and post-registration nursing curricula reflect both reductionist and holistic approaches and therefore cardiac nurses are suitably trained to manage reductionist as well as holistic care for clients with CHD. PMID- 17077776 TI - Telephone advice: advice given to a nursing home for pressure care. AB - This article considers the law which applies to the giving of advice over the telephone and the respective liability of the adviser and the recipient of the advice. It also covers the law relating to the exclusion of liability. PMID- 17077777 TI - Accelerating workforce redesign: threat or stimulus. PMID- 17077779 TI - "A tour of duty"... world nursing perspectives. PMID- 17077780 TI - A personal and ethical perspective on chronic wound pain. AB - Pain is a rarely acknowledged complication of chronic wounds, yet for those who live with it, it is often the over-riding preoccupation. In this article, the author--who herself has suffered for many years with wound pain--discusses why pain is not widely acknowledged by health professionals caring for wounds, and highlights why awareness of it is central to holistic care. PMID- 17077781 TI - Nurse case managers: patient care implications at a Pakistani university. AB - The role of the nurse in hospital is varied and some are choosing to incorporate more managerial and administrative skills into their clinical role. One such role is that of the nurse case manager (NCM). This particular role concentrates on involving the family and the patient in his or her own care, facilitation of the care plan, and open discussions between the patient, medics and nursing staff. NCMs in the author's hospital have made a remarkable contribution to patient care. It is a challenging and exceedingly demanding role in both developing and developed countries, but one that is increasingly important. The NCMs are involved in coordination, facilitation of core process and mobilization of resources, not only in hospital but at the patient's home. In this short introductory article the role of NCM is highlighted and the author discusses how this diverse role is concerned with patient care. NCMs work with multidisciplinary teams to enhance the patient's care process. Their attention is also given to cost reduction and clinical pathway management. PMID- 17077782 TI - Occupational back pain in Iranian nurses: an epidemiological study. AB - The aim of this study was to identify the prevalence and risk factors for low back pain (LBP) in nursing personnel and to analyze how individual and occupational characteristics contribute to the risk of LBP. Following ethical approval, 1226 nurses were randomly recruited from 13 general hospitals in northern Iran. Different questionnaires were designed to cover personal and professional data, the prevalence, and associations of risk factors with LBP. Results indicated that prevalence of LBP in nurses was over 50%. Lifting was the most common mechanism for LBP (30.4%). Prolonged standing and rest were found to be the significant aggravating and relieving factors (57.6% and 59.2%, respectively). Absence from work because of LBP in the month before the questionnaire was completed was reported by 33.7% of the sample. The results demonstrate that the magnitude of LBP among nursing personnel appears to be high and therefore more resources should be allocated to prevent such an injury occurring in the nursing profession. PMID- 17077783 TI - International prisoner health care: how nurses can do more. AB - This short article provides a summary account of the authors' recent visit to an international prisoner healthcare conference in Estonia, where there were a number of medical model presentations. The authors have explored the nursing contribution to prisoners' health care and have offered some examples of where this good practice is evident in England. The article highlights the value of nursing initiatives and how this should be given greater emphasis within international prisoner health care in the future. PMID- 17077784 TI - Nursing care delivery in Japanese psychiatric units. AB - Japan has the largest number of psychiatric beds per population in the world. At the same time, little research had been done to describe nursing care delivered in psychiatric units. To describe contents of nursing care, the framework for care classification and measuring methods must be examined. The purpose of this study was to develop a nursing intervention classification and examine the correlation between amount of care and patients' mental states. Data were collected from 39 patient care records from 4 randomly selected psychiatric hospitals. Patients' function and condition were evaluated by their doctors using the Global Assessment of Functioning (GAF) and the Brief Psychiatric Rating Scale (BPRS). Six care categories were extracted: basic activities of daily living, psychological support/interpersonal skills, medical support, monitoring, family support and miscellaneous. By examining three cases with this classification, there were significant correlations between the GAF and BPRS scores and amount of care for two acutely ill psychotic patients. PMID- 17077785 TI - Integrating simulation training into the nursing curriculum. AB - The use of simulation is gaining momentum in nurse education across the UK. The Nursing and Midwifery Council is currently investigating the use of simulation in pre-registration nursing. This article gives a brief history of simulation, discusses competence issues and why simulation is best placed to teach nurses in today's health service. An innovative approach to implementing simulation into the nursing curriculum is introduced. PMID- 17077786 TI - Centres of Excellence for the care of people with progressive ataxias. AB - This article provides a description of the progressive ataxias and how such a diagnosis impacts on a person's life. The nurse's role in helping these patients, who can face numerous difficulties in everyday life, is highlighted. Moreover, the authors discuss the development of Ataxia Specialist Centres of Excellence for the diagnosis and management of people with ataxia, and the benefits believe these centres will bring to the care of these patients. PMID- 17077787 TI - From spirituality to coping strategy: making sense of chronic illness. AB - This article explores how individuals might make sense of chronic illness. The spiritual aspect of self is described both as being central to finding meaning in suffering with a chronic illness and also the source of hope in meeting the challenges faced. Culture as the template for interpreting the significance of chronic ill health at a personal, familial and societal level is also considered. A conceptual model for understanding life transitions is modified to incorporate the spiritual and cultural perspectives of making sense of chronic illness in relation to coping skills. In understanding how patients make sense of their circumstances nurses are more likely to be able to offer appropriate support to effect coping. PMID- 17077788 TI - An introduction to thoracic surgery: assessment, diagnostics, treatment. AB - Thoracic surgery is an exciting and dynamic specialty. Many of the conditions treated can be regarded as life-limiting so practitioners require expert skills in communication as well as knowledge of the specific illnesses and complications that can arise. This article explains the types of surgery performed and discusses the factors that need to be taken into consideration before operating on malignant lung disease. The specific post-operative care required by patients following thoracic surgery is also discussed. PMID- 17077789 TI - A comparison of two methods of blood pressure measurement. AB - In current practice, a two-stage approach to measuring blood pressure (BP) has been widely accepted as the most accurate and reliable method. However, by changing the local haemodynamics, this procedure might alter the blood pressure. In a study of 39 subjects, blood pressure was measured using two indirect methods (two-stage and one-stage approaches). Results showed no statistically significant difference in values for systolic blood pressure obtained from the two methods. Statistically significant lower diastolic blood pressure values were obtained using the two-stage compared to the one-stage approach. It is proposed that initial inflation of the cuff to estimate systolic blood pressure in the two stage approach might lead to reactive hyperaemia and, therefore, a lower diastolic value. This two-stage approach might not provide the accurate readings it claims, and in addition it requires more time and subjects the patient to longer periods of stress. PMID- 17077790 TI - IV therapy: nurses must be competent to perform new duties. AB - This article considers the professional duty of the nurse to keep up to date and the extent, if any, to which she could refuse to take on expanded role activities, which in the past have normally been undertaken by other health professionals. It also considers the implications of the role of the new healthcare post of registered medical care practitioner. PMID- 17077791 TI - Developing older people's champions. PMID- 17077793 TI - Palliative care and education in Uganda. PMID- 17077794 TI - Supportive and palliative care in end stage renal failure: the need for further research. AB - In the UK, government guidelines related to chronic illness are being used to enhance supportive care of renal patients. Good practice includes ensuring the views of patients and carers are taken into account when developing and evaluating palliative care services. The literature relating to a renal patient's decision not to dialyse is sparse and the evidence base upon which to support effective nursing practice is currently weak. Much can be drawn from the discipline of cancer nursing, however, further research needs to be undertaken to develop new knowledge within the realm of supportive renal care. Having a fuller understanding of how, and when, renal patients finally make difficult end-of-life decisions is crucial in order to offer effective support and deliver an efficient and valuable renal supportive care service. PMID- 17077795 TI - Palliative care and support for people with neurodegenerative conditions and their carers. AB - AIM: to identify and compare the needs for supportive care/palliative care services of people in Australia with Motor Neurone Disease (MND), Multiple Sclerosis (MS), Huntington's Disease (HD) and Parkinson's Disease (PD) and the needs of their carers. DESIGN: national, descriptive survey of service use and support needs for 503 patients and 373 carers. Patients and carers were recruited from the databases of the disease associations in three Australian states. Patients were excluded if recently diagnosed or too sick or disabled to complete the survey. This was a specification of the ethics committee overseeing this project. FINDINGS: the MND and HD groups had the greatest needs for support and appeared most receptive to a palliative approach. The MS group was least likely to have a carer and more likely to live alone. HD and PD groups had the highest rates of anxiety and HD groups had the highest rate of depression. CONCLUSIONS: patients and carers who received more tailored services and more palliative care services were the most satisfied. Therefore, results highlight the need for tailored and flexible models of care for these groups with unique care requirements. PMID- 17077796 TI - Characteristics and views of family carers of older people with heart failure. AB - AIMS: to explore the characteristics and views of the family carers of older people with heart failure. METHOD: 213 family carers of heart failure patients >60 years were recruited from UK general practitioner (GP) practices. Carer strain, quality of life (QOL) and service satisfaction questionnaires were completed every 3 months for 2 years, as well as 16 interviews with patients and carers, and 9 focus groups with health care professionals. RESULTS: 76% of carers were female, 70% were >60 years and 73% were spousal carers. Predictors of carer strain were symptoms of depression, age and patient NYHA. Predictors of lower QOL were: spousal carer; 2+ health conditions and symptoms of depression. Qualitative findings related to the change in circumstances, impact of responsibilities and health conditions of the family carers. CONCLUSION: carers were mainly older women, often experiencing multiple health conditions. Addressing the practical and emotional support required presents a challenge for specialist palliative care in responding to calls for increased involvement in heart failure. A shared care model with liaison between specialist heart failure nurses, cardiologists, primary care teams and hospice services is advocated. PMID- 17077797 TI - People with chronic obstructive pulmonary disease at the end of life: a review of the literature. AB - Based on 2004 data, chronic obstructive pulmonary disease (COPD) is the fourth leading cause of death in the world, surpassed only by cardiovascular disease, pneumonia and HIV/AIDS. The terminal trajectory of patients with COPD is distinct from that of cancer patients. The unpredictability of prognosis for people with COPD poses different challenges in end-of-life decision-making from those faced by individuals with terminal cancer. The use of a traditional cancer-based service model to predict the need for palliative care services is not helpful for people with COPD. Drastic improvements in end of life care for the people with COPD are essential, especially with the projected rise in cases over the coming years. PMID- 17077798 TI - Taking on a critical new role for hospices. PMID- 17077799 TI - Towards a definition of death anxiety. AB - There is currently no consensus about what the notion of death anxiety means. This commentary explores the complexity of death anxiety, asserts the distinction between anxiety and fear, and attempts a definition that encompasses the many facets associated with the term. PMID- 17077800 TI - The Liverpool Care Pathway in hospices: an exploratory study of doctor and nurse perceptions. AB - BACKGROUND: The Liverpool Care of the Dying Pathway (LCP) was developed to transfer the hospice model of care into other care settings. It is a multiprofessional document providing an evidence-based framework for the dying phase. Widely used in hospitals it has now been adopted for use in hospices. METHOD: A qualitative study exploring hospice doctors' and nurses' perceptions of the impact of the LCP using semi-structured interviews of a purposive sample were undertaken in a 30 bed hospice in the north-west of England. Data was thematically analysed for emerging themes. RESULTS AND DISCUSSION: Eleven interviews were undertaken (eight nurses, three doctors). The results suggest that despite some initial scepticism the LCP has a valuable place in the hospice setting according to the doctors and nurses sampled. Alongside the potential for use in audit and research, perceived benefits include improving documentation, promoting continuity of care and enhancing communication and the care of relatives. Maintaining ongoing education and feedback on the use of the document were highlighted as important mechanisms for ensuring that the delivery of care remained optimum. PMID- 17077801 TI - Respite care for children, young people and families--are their needs addressed? AB - In recent years there has been an increasing demand for respite care in the community setting, as advances in medicine and technology have enhanced the life expectancy of children and young people with a life-limiting illness. However, the need for flexible respite provision in the home frequently presents a challenge for those health care professionals involved in caring for these children and their families. Following a review of literature this paper will demonstrate how a family-centred approach to respite provision and palliative care can enhance the quality of life for a child nearing the end of life and support the expressed needs of the child's family with accessible respite care. PMID- 17077802 TI - Adult bereavement in five English hospices: types of support. AB - METHOD: In-depth organisational case studies of five English hospices. ANALYSIS: Thematic analysis of qualitative interviews and focus groups and scrutiny of documentary material provided by the hospice bereavement services. FINDINGS: All of the hospices ran events where bereaved people met with other bereaved people who had been in contact with the hospice. They all offered some form of more intensive one-to-one support, spiritual support, and ran remembrance events. Three types of one-to-one support were offered: counselling; befriending; support from paid bereavement staff. Ongoing telephone support from trained bereavement volunteers appeared to be an acceptable and cost-effective way of providing low intensity bereavement support. Running social bereavement support groups is difficult and resource intensive. "Drop-in" events appear to perform a useful role for both bereaved people and bereavement services. There seemed to be no readily available sources of support for people with particularly complex bereavement problems. CONCLUSIONS: All of the hospices were offering appropriate types of bereavement support that clearly met Components 1 and 2 of bereavement support in the NICE guidance. In the absence of any agreement about "best practice" for adult bereavement support services it is the integration of bereavement support as a central aspect of hospice activity that is most likely to improve bereavement support for adults in the future. PMID- 17077803 TI - Canada's Compassionate Care Benefit: views of family caregivers in chronic illness. AB - AIM: Based on a pilot evaluation of Canada's recently introduced Compassionate Care Benefit (CCB), the purpose of this paper is to highlight the experiences of family caregivers caring for people with non-malignant advanced chronic illness. DESIGN: Using Patton's (1997) utilization-focused evaluation approach, 25 telephone interviews were conducted with three groups of family caregivers: those who had successfully applied for the CCB; those who were unsuccessful in their applications; those who had never applied for the benefit. FINDINGS: The CCB has a number of limitations, particularly for caregivers of patients diagnosed with non-malignant advanced chronic illness. The central limitations are: difficulties associated with accurate prognostication; limited definition of "family member"; insufficient length of the funding period. CONCLUSION: By modelling similar programmes internationally, such as those in Sweden, Norway, and the Netherlands, Canada would likely find the CCB to have greater relevance and accessibility to Canadian caregivers, particularly those caring for people with non-malignant advanced chronic illness. PMID- 17077805 TI - Long-term outcome among men with conservatively treated localised prostate cancer. AB - Optimal management of clinically localised prostate cancer presents unique challenges, because of its highly variable and often indolent natural history. There is an urgent need to predict more accurately its natural history, in order to avoid unnecessary treatment. Medical records of men diagnosed with clinically localised prostate cancer, in the UK, between 1990 and 1996 were reviewed to identify those who were conservatively treated, under age 76 years at the time of pathological diagnosis and had a baseline prostate-specific antigen (PSA) measurement. Diagnostic biopsy specimens were centrally reviewed to assign primary and secondary Gleason grades. The primary end point was death from prostate cancer and multivariate models were constructed to determine its best predictors. A total of 2333 eligible patients were identified. The most important prognostic factors were Gleason score and baseline PSA level. These factors were largely independent and together, contributed substantially more predictive power than either one alone. Clinical stage and extent of disease determined, either from needle biopsy or transurethral resection of the prostate (TURP) chips, provided some additional prognostic information. In conclusion, a model using Gleason score and PSA level identified three subgroups comprising 17, 50, and 33% of the cohort with a 10-year prostate cancer specific mortality of <10, 10-30, and >30%, respectively. This classification is a substantial improvement on previous ones using only Gleason score, but better markers are needed to predict survival more accurately in the intermediate group of patients. PMID- 17077807 TI - Interactions between nitric oxide and corticosterone in the regulation of progenitor cell proliferation in the dentate gyrus of the adult rat. AB - It is well established that L-NAME, a generic NOS inhibitor, stimulates neurogenesis in the dentate gyrus of the adult rat and corticosterone reduces it. These experiments explore the interaction between L-NAME and corticosterone. L NAME (50 mg/kg), as expected, increased proliferation, but also lowered plasma corticosterone levels. However, the stimulating action of L-NAME depends on the presence of rhythmic changes in plasma corticosterone, as it is abolished in rats treated with a subcutaneous implant of corticosterone, which flattens the diurnal rhythm. Adrenalectomized rats implanted with corticosterone also failed to respond to L-NAME. Giving them a single daily injection of corticosterone (2 mg/kg) in an attempt to replicate the diurnal rhythm restored the sensitivity of the progenitor cells to L-NAME. The mechanism for this result remains to be investigated. Excess corticosterone given by daily injection (40/mg/kg) reduced proliferation but did not alter the response to L-NAME, even though this occurred from a lower baseline. nNOS was demonstrable only in the inner (proliferative) layer of the dentate gyrus in control rats, and did not alter following excess corticosterone treatment. iNOS was detectable at low levels in control rats, but was increased markedly following corticosterone. eNOS was evident throughout the dentate gyrus, and also increased after corticosterone (particularly in the hilus). Aminoguanidine (100 mg/kg/day; an iNOS antagonist) significantly increased proliferation in corticosterone-treated rats (40 mg/kg/day) but not in controls without additional corticosterone, confirming that iNOS plays a role in corticosterone-regulated neurogenesis. Corticosterone may thus act on progenitor cells in part at least through increased nitric oxide (NO) formation. The effects of reduced NO on neurogenesis may rely on a dual mechanism: corresponding reductions in plasma corticosterone and increased induction of iNOS (and/or eNOS) within the dentate gyrus. The possibility that NO acts downstream of glucocorticoids in the dentate gyrus is suggested. PMID- 17077808 TI - Association of 4-repeat allele of the dopamine D4 receptor gene exon III polymorphism and response to methylphenidate treatment in Korean ADHD children. AB - In the present study, we investigated the association between the 4-repeat allele at the dopamine receptor D4 (DRD4) gene and the response to treatment with methylphenidate (MPH) in Korean children with attention deficit hyperactivity disorder (ADHD). The study subjects were 83 children with ADHD (8.40+/-1.73 years) who were recruited from two child psychiatric clinics in South Korea. All of the drug-naive ADHD children were treated with MPH for about 8 weeks. An improvement of more than or equal to [corrected] 50% in the ADHD Rating Scale-IV (ARS) scores after 8 weeks of treatment compared with the baseline ARS scores before the treatment was considered as a 'good response', whereas an improvement of less than [corrected] 50% was considered as a 'poor response'. After the genotyping for DRD4 was performed, we investigated the association between the genotype at DRD4 and the response to MPH treatment. We performed a comparison of the response to MPH treatment between the two largest groups, viz. the subjects with and without the 4/4 genotype at DRD4. According to the ARS scores of the subjects as assessed by their parents and by their teachers, we found that while 71.1 and 80.0% (32/45 and 24/30), respectively, of those with a good response to MPH treatment showed the 4/4 genotype at DRD4, only 31.6 and 37.7% (12/38 and 20/53), respectively, of those with a poor response to MPH treatment showed the 4/4 genotype at DRD4 (Pearson chi2-values=12.926 and 13.737, respectively, both df=1, and both p<0.01). Our findings support the existence of an association between the 4-repeat allele at DRD4 and good response to MPH in Korean ADHD children. PMID- 17077809 TI - Striatal vs extrastriatal dopamine D2 receptors in antipsychotic response--a double-blind PET study in schizophrenia. AB - Blockade of dopamine D2 receptors remains a common feature of all antipsychotics. It has been hypothesized that the extrastriatal (cortical, thalamic) dopamine D2 receptors may be more critical to antipsychotic response than the striatal dopamine D2 receptors. This is the first double-blind controlled study to examine the relationship between striatal and extrastriatal D2 occupancy and clinical effects. Fourteen patients with recent onset psychosis were assigned to low or high doses of risperidone (1 mg vs 4 mg/day) or olanzapine (2.5 mg vs 15 mg/day) in order to achieve a broad range of D2 occupancy levels across subjects. Clinical response, side effects, striatal ([11C]-raclopride-positron emission tomography (PET)), and extrastriatal ([11C]-FLB 457-PET) D2 receptors were evaluated after treatment. The measured D2 occupancies ranged from 50 to 92% in striatal and 4 to 95% in the different extrastriatal (frontal, temporal, thalamic) regions. Striatal and extrastriatal occupancies were correlated with dose, drug plasma levels, and with each other. Striatal D2 occupancy predicted response in positive psychotic symptoms (r=0.62, p=0.01), but not for negative symptoms (r=0.2, p=0.5). Extrastriatal D2 occupancy did not predict response in positive or negative symptoms. The two subjects who experienced motor side effects had the highest striatal occupancies in the cohort. Striatal D2 blockade predicted antipsychotic response better than frontal, temporal, and thalamic occupancy. These results, when combined with the preclinical data implicating the mesolimbic striatum in antipsychotic response, suggest that dopamine D2 blockade within specific regions of the striatum may be most critical for ameliorating psychosis in schizophrenia. PMID- 17077810 TI - Chronic treatment with desipramine improves cognitive performance of rats in an attentional set-shifting test. AB - Alterations in central monoaminergic neurotransmission are important in the actions of many antidepressants. This study tested the hypothesis that tonic elevation of noradrenergic (NA) neurotransmission in medial prefrontal cortex (mPFC) by chronic treatment with the selective norepinephrine (NE) reuptake blocker desipramine (DMI) may contribute to the beneficial cognitive effects of this antidepressant drug (AD). Male Sprague-Dawley rats were treated with DMI acutely (15 mg/kg, i.p.) or chronically for 21 days (7.5 mg/kg/day via osmotic minipump) before assessing performance on an attentional set-shifting test. The extradimensional set-shifting component of this test reflects a process of cognitive flexibility that is dependent upon mPFC, and that we have shown previously to be facilitated by NA activity in mPFC. Microdialysis was performed to measure NE release in mPFC concurrently with behavioral testing. Acute DMI treatment produced an increase in extracellular NE levels in mPFC, and a modest improvement in overall performance across all task stages of the attentional set shifting test, but failed to produce a significant improvement in any of the individual specific tasks comprising the test sequence. Chronic DMI treatment tonically elevated basal extracellular NE levels in mPFC, associated with a significant improvement in performance specifically on the extradimensional set shifting component of the test. There was also a significant reduction in set loss errors in rats treated chronically with DMI. Hence, tonic elevation of NA transmission in mPFC by chronic DMI treatment was associated with a time dependent facilitation of cognitive flexibility that may contribute to the mechanism whereby chronic treatment with ADs, specifically NE reuptake blockers, may exert a beneficial therapeutic effect on cognition in depressed patients. PMID- 17077811 TI - FG 7142 specifically reduces meal size and the rate and regularity of sustained feeding in female rats: evidence that benzodiazepine inverse agonists reduce food palatability. AB - Benzodiazepine receptor inverse agonists reduce food intake in males, but their actions in females, in whom stress-related eating disorders are more common, as well as their behavioral mode of action remain unclear. The consummatory effects of benzodiazepine receptor ligands have alternately been hypothesized to reflect changes in the hedonic evaluation of food or secondary effects of anxiety-related or cognitive properties. To test the anorectic mode of action of benzodiazepine inverse agonists, the effects of FG 7142 on feeding microstructure were studied in nondeprived female Wistar rats (n=32). Microstructure analysis used a novel meal definition that recognizes prandial drinking. On pharmacologically synchronized diestrus I, rats were pretreated (-30 min dark onset) with the benzodiazepine partial inverse agonist FG 7142 (i.p. 0, 3.75, 7.5, 15 mg/kg) in a between-subjects design. FG 7142 delayed the onset of (16-541%), decreased the amount eaten (36-52%) and drunk (63-87%), and reduced the time spent drinking (59 87%) within the first nocturnal meal. Dose-dependent incremental anorexia continued 6 h into the dark cycle, whereas FG 7142 did not suppress the quantity, duration or rate of drinking past the first meal. Treated rats ate smaller meals (17-42%) of normal duration. This reflected that FG 7142 slowed feeding within meals (9-38%) by decreasing the regularity and maintenance of feeding from pellet to-pellet. FG 7142 did not influence postprandial satiety; meal frequency and inter-meal intervals were unaffected. FG 7142 anorexia was blocked by the benzodiazepine receptor antagonist flumazenil in a 2:1 molar ratio (n=17 rats). The very early, nonspecific (+10 min), but not subsequent (2.5, 4.5 h) feeding specific phase, of FG 7142 anorexia was mirrored by anxiogenic-like behavior in FG 7142-treated (7.5 mg/kg) female rats (n=48) in the elevated plus-maze. Thus, benzodiazepine receptor inverse agonists preferentially lessen the maintenance of feeding in female rats, effects opposite to those of palatable food. PMID- 17077812 TI - A prospective cohort study on sustained effects of low-dose ecstasy use on the brain in new ecstasy users. AB - It is debated whether ecstasy use has neurotoxic effects on the human brain and what the effects are of a low dose of ecstasy use. We prospectively studied sustained effects (>2 weeks abstinence) of a low dose of ecstasy on the brain in ecstasy-naive volunteers using a combination of advanced MR techniques and self report questionnaires on psychopathology as part of the NeXT (Netherlands XTC Toxicity) study. Outcomes of proton magnetic resonance spectroscopy (1H-MRS), diffusion tensor imaging (DTI), perfusion-weighted imaging (PWI), and questionnaires on depression, impulsivity, and sensation seeking were compared in 30 subjects (12M, 21.8+/-3.1 years) in two sessions before and after first ecstasy use (1.8+/-1.3 tablets). Interval between baseline and follow-up was on average 8.1+/-6.5 months and time between last ecstasy use and follow-up was 7.7+/-4.4 weeks. Using 1H-MRS, no significant changes were observed in metabolite concentrations of N-acetylaspartate (NAA), choline (Cho), myo-inositol (mI), and creatine (Cr), nor in ratios of NAA, Cho, and mI relative to Cr. However, ecstasy use was followed by a sustained 0.9% increase in fractional anisotropy (FA) in frontoparietal white matter, a 3.4% decrease in apparent diffusion (ADC) in the thalamus and a sustained decrease in relative regional cerebral blood volume (rrCBV) in the thalamus (-6.2%), dorsolateral frontal cortex (-4.0%), and superior parietal cortex (-3.0%) (all significant at p<0.05, paired t-tests). After correction for multiple comparisons, only the rrCBV decrease in the dorsolateral frontal cortex remained significant. We also observed increased impulsivity (+3.7% on the Barratt Impulsiveness Scale) and decreased depression ( 28.0% on the Beck Depression Inventory) in novel ecstasy users, although effect sizes were limited and clinical relevance questionable. As no indications were found for structural neuronal damage with the currently used techniques, our data do not support the concern that incidental ecstasy use leads to extensive axonal damage. However, sustained decreases in rrCBV and ADC values may indicate that even low ecstasy doses can induce prolonged vasoconstriction in some brain areas, although it is not known whether this effect is permanent. Additional studies are needed to replicate these findings. PMID- 17077813 TI - Postischemic gene transfer of soluble Flt-1 protects against brain ischemia with marked attenuation of blood-brain barrier permeability. AB - Brain edema is a major and often mortal complication of brain ischemia. Vascular endothelial growth factor (VEGF) is also known as a potent vascular permeability factor and may play detrimental roles at the acute stage of brain infarction. Our goal in this study was to explore protective effects of gene transfer of soluble flt-1 (sFlt-1), a natural inhibitor of VEGF, on focal brain ischemia. Adenoviral vector encoding sFlt-1 or beta-galactosidase as control was injected into the lateral ventricle 90 mins after photochemical distal middle cerebral artery occlusion in male spontaneously hypertensive rats. The transduced sFlt-1 was released to the cerebrospinal fluid from the ventricular wall and significantly increased 6 h, 1 and 7 days after sFlt-1 transfection. One day after brain ischemia, sFlt-1 gene transfer significantly reduced infarct volume (by 35%), brain edema (by 35%), and blood-brain barrier permeability (Evans blue extravasation; by 69%) with diminished phosphorylation of focal adhesion kinase (FAKtyr397 and FAKtyr861) in the ischemic vessels. Seven days after ischemia, sFlt-1 gene transfer also significantly attenuated infarct volume (by 29%) and monocyte/macrophage infiltration (by 27%), although there were no reductions in angiogenesis by sFlt-1 overexpression. These results suggest that sFlt-1 gene therapy targeting brain edema in acute stage of brain ischemia may be useful for brain infarction. PMID- 17077814 TI - Transport pathways for clearance of human Alzheimer's amyloid beta-peptide and apolipoproteins E and J in the mouse central nervous system. AB - Amyloid beta-peptide (Abeta) clearance from the central nervous system (CNS) maintains its low levels in brain. In Alzheimer's disease, Abeta accumulates in brain possibly because of its faulty CNS clearance and a deficient efflux across the blood-brain barrier (BBB). By using human-specific enzyme-linked immunosorbent assays, we measured a rapid 30 mins efflux at the BBB and transport via the interstitial fluid (ISF) bulk flow of human-unlabeled Abeta and of Abeta transport proteins, apolipoprotein E (apoE) and apoJ in mice. We show (i) Abeta40 is cleared rapidly across the BBB via low-density lipoprotein receptor-related protein (LRP)1 at a rate of 0.21 pmol/min g ISF or 6-fold faster than via the ISF flow; (ii) Abeta42 is removed across the BBB at a rate 1.9-fold slower compared with Abeta40; (iii) apoE, lipid-poor isoform 3, is cleared slowly via the ISF flow and across the BBB (0.03-0.04 pmol/min g ISF), and after lipidation its transport at the BBB becomes barely detectable within 30 mins; (iv) apoJ is eliminated rapidly across the BBB (0.16 pmol/min g ISF) via LRP2. Clearance rates of unlabeled and corresponding 125I-labeled Abeta and apolipoproteins were almost identical, but could not be measured at low physiologic levels by mass spectrometry. Amyloid beta-peptide 40 binding to apoE3 reduced its efflux rate at the BBB by 5.7-fold, whereas Abeta42 binding to apoJ enhanced Abeta42 BBB clearance rate by 83%. Thus, Abeta, apoE, and apoJ are cleared from brain by different transport pathways, and apoE and apoJ may critically modify Abeta clearance at the BBB. PMID- 17077815 TI - Erythropoietin-induced neurovascular protection, angiogenesis, and cerebral blood flow restoration after focal ischemia in mice. AB - Restoration of local blood supply in the post-ischemic brain plays a critical role in tissue repair and functional recovery. The present investigation explored beneficial effects of recombinant human erythropoietin (rhEPO) on vascular endothelial cell survival, angiogenesis, and restoration of local cerebral blood flow (LCBF) after permanent focal cerebral ischemia in adult mice. Saline or rhEPO (5,000 U/kg, intraperitoneal) was administered 30 mins before ischemia and once daily after ischemic stroke. Immunohistochemistry showed an enhancing effect of rhEPO on expression of EPO receptor (EPOR) of endothelial cells in the penumbra region 3 to 21 days after the ischemic insult. The treatment with rhEPO decreased ischemia-induced cell death and infarct volume 3 days after stroke. Specifically, rhEPO reduced the number of terminal deoxynucleotidyl transferase biotin-dUPT nick end labeling- and caspase-3-positive endothelial cells in the penumbra region. Colocalization of the vessel marker glucose transporter-1 (Glut 1) and cell proliferation marker 5-bromo-2'-deoxyuridine indicated enhanced angiogenic activity in rhEPO-treated mice 7 to 21 days after stroke. Western blot showed upregulation of the expression of angiogenic factors Tie-2, Angiopoietin 2, and vascular endothelial growth factor in rhEPO-treated animals. Local cerebral blood flow was measured by laser scanning imaging 3 to 21 days after stroke. At 14 days, LCBF in the penumbra was recovered to preischemia levels in rhEPO-treated mice but not in control mice. Our data suggest that rhEPO treatment upregulates the EPOR level in vascular endothelial cells, confers neurovascular protection, and enhances angiogenesis. We further show a promoting effect of rhEPO on LCBF recovery in the ischemic brain. These rhEPO-induced effects may contribute to therapeutic benefits in the treatment of ischemic stroke. PMID- 17077816 TI - Capillary-oxygenation-level-dependent near-infrared spectrometry in frontal lobe of humans. AB - Brain function requires oxygen and maintenance of brain capillary oxygenation is important. We evaluated how faithfully frontal lobe near-infrared spectroscopy (NIRS) follows haemoglobin saturation (SCap) and how calculated mitochondrial oxygen tension (PMitoO2) influences motor performance. Twelve healthy subjects (20 to 29 years), supine and seated, inhaled O2 air-mixtures (10% to 100%) with and without added 5% carbon dioxide and during hyperventilation. Two measures of frontal lobe oxygenation by NIRS (NIRO-200 and INVOS) were compared with capillary oxygen saturation (SCap) as calculated from the O2 content of brachial arterial and right internal jugular venous blood. At control SCap (78%+/-4%; mean+/-s.d.) was halfway between the arterial (98%+/-1%) and jugular venous oxygenation (SvO2; 61%+/-6%). Both NIRS devices monitored SCap (P<0.001) within approximately 5% as SvO2 increased from 39%+/-5% to 79%+/-7% with an increase in the transcranial ultrasound Doppler determined middle cerebral artery flow velocity from 29+/-8 to 65+/-15 cm/sec. When SCap fell below approximately 70% with reduced flow and inspired oxygen tension, PMitoO2 decreased (P<0.001) and brain lactate release increased concomitantly (P<0.001). Handgrip strength correlated with the measured (NIRS) and calculated capillary oxygenation values as well as with PMitoO2 (r>0.74; P<0.05). These results show that NIRS is an adequate cerebral capillary-oxygenation-level-dependent (COLD) measure during manipulation of cerebral blood flow or inspired oxygen tension, or both, and suggest that motor performance correlates with the frontal lobe COLD signal. PMID- 17077824 TI - Tackling investigator noncompliance: Suspend the surgeries. PMID- 17077825 TI - Tackling investigator noncompliance. PMID- 17077826 TI - Tackling investigator noncompliance: Cooperation counts. PMID- 17077827 TI - Tackling investigator noncompliance: Decide now. PMID- 17077828 TI - Tackling investigator noncompliance: Stop and investigate. PMID- 17077829 TI - Tackling investigator noncompliance: a word from OLAW and USDA. PMID- 17077830 TI - Fecal bulking in a frequently mated female rat. Colonic obstruction due to severe vaginal distension. PMID- 17077832 TI - The new transgenic mouse core: no longer a facility just for making mice. AB - The times they are a-changin' for GEM and the facilities that maintain them. Core facilities are expanding beyond their original conception as producers of transgenic mice to encompass a wide range of services, including research animal maintenance. In this paper, the authors describe the logistics and administration of the newly dubbed Mouse Genetics Core Facility at the Memorial Sloan-Kettering Cancer Center as a blueprint for other institutions seeking to expand and update their own transgenic cores for research in the twenty-first century. PMID- 17077833 TI - An interdisciplinary performance-based approach to training laboratory animal technicians. AB - Experience has proven that comprehensive training and education make it easier to attract and retain highly qualified animal care technicians, as well as to ensure that research facilities reach maximum performance. The authors outline the training approach used at SoBran, Inc., by describing the subject matter covered in their initial orientation period and in ongoing training sessions. The authors also address recordkeeping methods and training-program assessment. PMID- 17077834 TI - Spontaneous otitis media in Wistar rats: an overlooked pathology in otological research. AB - The rat is commonly employed in otological research, but spontaneous ear infections can confound the results of experimental procedures--wasting time, money, and animals. The authors focus on the incidence of spontaneous otitis media in Wistar rats. They compare disease incidence in animals housed in standard cages with those housed in barrier units, showing that 20% of their conventionally housed animals developed spontaneous otitis media, whereas only 5% of their animals housed in isolated units were infected. These results underscore the importance of strict control of the shipping, housing conditions, and manipulation of animals to be used in otological research. PMID- 17077837 TI - Sail through those interviews. PMID- 17077838 TI - European competition. PMID- 17077839 TI - Dislocation propagation versus dislocation nucleation. PMID- 17077840 TI - Dislocation propagation versus dislocation nucleation. PMID- 17077841 TI - Universal tools for biomolecular attachment to surfaces. PMID- 17077842 TI - Nurturing science in developing countries. PMID- 17077844 TI - Computational methods: a search engine for catalysts. PMID- 17077846 TI - Electrowetting: shake, rattle and roll. PMID- 17077845 TI - Correlated oxides: all aboard the manganite rollercoaster. PMID- 17077847 TI - Nanofabrication: best of both worlds. PMID- 17077848 TI - Quantum dots: artificial atoms for quantum optics. PMID- 17077850 TI - All systems go! PMID- 17077852 TI - ROS as a tumour suppressor? PMID- 17077853 TI - Ahead of the curve: mitochondrial fusion and phospholipase D. PMID- 17077854 TI - PERing in their sleep. PMID- 17077855 TI - Working out coupled monoubiquitination. PMID- 17077856 TI - SePARating polarity and proliferation in ErbB2 oncogenesis. PMID- 17077858 TI - What is a PhD? PMID- 17077859 TI - Food for tomorrow? How the scientific discipline of molecular gastronomy could change the way we eat. PMID- 17077860 TI - The return of Romeo. Scientists' international mobility and the future of research in Europe. PMID- 17077861 TI - Genetic genealogy goes global. Although useful in investigating ancestry, the application of genetics to traditional genealogy could be abused. PMID- 17077862 TI - Saving a fragile legacy. Biotechnology and microbiology are increasingly used to preserve and restore the world's cultural heritage. PMID- 17077863 TI - Food allergies on the rise? Determining the prevalence of food allergies, and how quickly it is increasing, is the first step in tackling the problem. PMID- 17077864 TI - Do orphan G-protein-coupled receptors have ligand-independent functions? New insights from receptor heterodimers. AB - G-protein-coupled receptors (GPCRs) are important drug targets and are involved in virtually every biological process. However, there are still more than 140 orphan GPCRs, and deciphering their function remains a priority for fundamental and clinical research. Research on orphan GPCRs has concentrated mainly on the identification of their natural ligands, whereas recent data suggest additional ligand-independent functions for these receptors. This emerging concept is connected with the observation that orphan GPCRs can heterodimerize with GPCRs that have identified ligands, and by so doing regulate the function of the latter. Pairing orphan GPCRs with their potential heterodimerization partners will have a major impact on our understanding of the extraordinary diversity offered by GPCR heterodimerization and, in addition, will constitute a novel strategy to elucidate the function of orphan receptors that needs to be added to the repertoire of 'deorphanization' strategies. PMID- 17077865 TI - The structure of the Sec complex and the problem of protein translocation. AB - Proteins synthesized in the cytosol either remain there or are localized to a specific membrane and subsequently translocated to another cellular compartment. These extracytosolic proteins have to cross, or be inserted into, a phospholipid bilayer-a process governed by membrane-bound protein transporters designed to recognize and receive appropriate polypeptides and thread them through the membrane. One such translocation complex, SecY/Sec61, is found in every cell, in either the plasma membrane of bacteria and archaea or the endoplasmic reticulum membrane of eukaryotes. Recent structural findings, combined with previous genetic and biochemical studies, have helped to describe how the passage of proteins through the membrane might occur, but several points of uncertainty remain. PMID- 17077868 TI - Simulation Procedure in Periodic Cancer Screening Trials. AB - A general simulation procedure is described to validate model fitting algorithms for complex likelihood functions that are utilized in periodic cancer screening trials. Although screening programs have existed for a few decades, there are still many unsolved problems, such as how age or hormone affects the screening sensitivity, the sojourn time in the preclinical state, and the transition probability from disease-free state to the preclinical state. Simulations are needed to check reliability or validity of the likelihood function combined with the associated effect functions. One bottleneck in the simulation procedure is the very time consuming calculations of the maximum likelihood estimates (MLE) from generated data. A practical procedure is presented, along with results for when both sensitivity and transition probability into the preclinical state are age-dependent. The procedure is also suitable for other applications. PMID- 17077866 TI - The role of synaptic ion channels in synaptic plasticity. AB - The nervous system receives a large amount of information about the environment through elaborate sensory routes. Processing and integration of these wide ranging inputs often results in long-term behavioural alterations as a result of past experiences. These relatively permanent changes in behaviour are manifestations of the capacity of the nervous system for learning and memory. At the cellular level, synaptic plasticity is one of the mechanisms underlying this process. Repeated neural activity generates physiological changes in the nervous system that ultimately modulate neuronal communication through synaptic transmission. Recent studies implicate both presynaptic and postsynaptic ion channels in the process of synapse strength modulation. Here, we review the role of synaptic ion channels in learning and memory, and discuss the implications and significance of these findings towards deciphering the molecular biology of learning and memory. PMID- 17077869 TI - The clinical characteristics of tinnitus in patients with vestibular schwannoma. AB - OBJECTIVES: To review the symptoms, signs, and clinical findings in a large series of patients diagnosed with unilateral sporadic vestibular schwannoma (VS) to describe the clinical characteristics of tinnitus in this population. Further, to ascertain which of the proposed mechanisms of tinnitus generation in VS was supported. DESIGN: Retrospective case note and database review. SETTING: Tertiary university teaching hospital departments of audiology and neuro-otology. PARTICIPANTS: Nine hundred forty-one patients with unilateral sporadic VS, diagnosed during the period 1986 to 2002. Twenty-three additional patients were excluded due to missing clinical data. MAIN OUTCOME MEASURES: The presence or absence of tinnitus, and its rated subjective severity were analyzed in conjunction with data regarding patient demographics, symptoms, signs, and diagnostic audiovestibular test findings. RESULTS: No statistical association at the 5% level was found between tinnitus presence/absence and patient age, gender, 2- to 4-kHz audiometric thresholds, ipsilateral auditory brainstem response abnormality, length of history, tumor side, nor caloric test abnormality. Statistically significant associations were found between tinnitus presence/absence and tumor size (p = 0.012) and type of hearing loss (progressive, sudden, fluctuant, nil) with a tendency for patients without hearing loss to be less likely to experience tinnitus. Statistically significant associations were identified between classification of tinnitus severity and age at diagnosis (p < 0.001) (greater age being associated with greater tinnitus severity), abnormal findings on caloric testing (p = 0.01) (abnormal calorics being associated with greater tinnitus severity), and tinnitus as a principal presenting symptom (p < 0.001) (this being associated with greater tinnitus severity). CONCLUSIONS: The analysis does not identify any single one of the proposed mechanisms for tinnitus as being the obvious culprit. In fact, even in a homogeneous group of patients such as this, there is evidence of multiple mechanisms that are not mutually exclusive. The association between increased tinnitus severity in older patients, patients with canal pareses on caloric testing, and with tinnitus as a principal presenting symptom should be borne in mind by the clinician. PMID- 17077870 TI - Virtual reality augmentation in skull base surgery. AB - OBJECTIVE: Skull base anatomy is complex and subject to individual variation. Understanding the complexity of surgical anatomy is faster and easier with virtual models created from primary imaging data of the patient. This study was designed to investigate the usefulness of virtual reality in image guidance for skull base procedures. DESIGN: Primary volumetric image data from 110 patients was acquired using magnetic resonance, computed tomography (CT), and CT angiography. Pathologies included lesions in the anterior, middle, and posterior skull base. The data were transferred to an infrared-based image-guidance system for creation of a virtual operating field (VOF) with translucent surface modulation and optional "fly-through" video mode. During surgery, the target registration error for anatomical landmarks was assessed and the VOF was compared with the patient's anatomy in the operative field. RESULTS: Complex structures like the course of the sigmoid sinus, the carotid artery, and the outline of the paranasal sinuses were well visualized in the VOF and were recognized by the surgeon instantly. Perception was greatly facilitated as compared with routine mental reconstruction of triaxial images. Accurate assessment of the depth of field and very small objects was not possible in VOF images. CONCLUSION: Supported by sound anatomical knowledge, creation of a virtual operating field for a surgical approach in an individual patient offers a deja vu experience that can enhance the capabilities of a surgical team in skull base approaches. In addition, application of this technique in image-guided procedures assists in targeting or avoiding hidden anatomical structures. PMID- 17077871 TI - Unusual tumors of the posterior fossa skull base. AB - Five unusual cases of posterior skull base tumors were treated through different skull base approaches. Two or more staged operations were required to achieve total or near-total excision and decompression of two extensive tumors. Total excision of an extensive en plaque meningioma of the foramen magnum that encircled the brain stem and cervical spinal cord could not be achieved through the extreme lateral and suboccipital craniectomy approach. However, the vital structures were decompressed and the patient's postoperative morbidity was acceptable. An extended middle fossa approach was required to excise a hemangiopericytoma of the middle and posterior fossae in a 12-year-old. Extension of the tumor into the posterior fossa precluded a retromastoid approach because the mass draped the lower cranial nerves posteriorly. Two men had undifferentiated adenocarcinomas involving the jugular foramen and middle to posterior fossa, respectively. The origin of one was renal in a 37-year-old man. A 63-year-old man survived 1.5 years after a good decompression of his extensive tumor and irradiation. The histological diagnosis of paraganglioma of the occipital bone was a surprise in a 25-year-old man with pure bony involvement. These cases indicate that the appropriate selection of skull base approaches and their combination can provide the needed access to achieve adequate excision or decompression of masses located in challenging anatomical regions of the skull base. Furthermore, good surgical excision improves palliation in radioresistant metastatic tumors of the skull base. PMID- 17077872 TI - Surgical management of petroclival chordomas: report of eight cases. AB - In the management of skull base chordomas, surgical treatment is essential to achieve long-term control. A petroclival chordoma growing laterally in the skull base is one of the most challenging tumors for neurosurgeons. We have treated petroclival chordomas based on the principle of maximal surgical resection of the tumor with minimal morbidity. Lateral skull base approaches were used to approach petroclival chordomas in eight patients. The surgical procedure involved removal of soft tumor tissue and extensive drilling of adjacent bony structures. Gross total resection of the tumor was achieved in six patients. Subtotal resection in the remaining two patients was associated with acceptable morbidity. In cases of petroclival chordomas, lateral skull base approaches can be used as a primary procedure, although those approaches may be associated with high rates of morbidity and mortality. PMID- 17077873 TI - Conservative management of acoustic neuroma. AB - AIM OF STUDY: To identify those patients with vestibular schwannoma (acoustic neuroma) in whom treatment becomes necessary. METHOD: Retrospective chart review. RESULT: A total of 205 patients with small tumors were followed for a mean of 40.8 months. The longest follow-up was 180 months. One hundred and ninety-seven patients had a follow-up of more than 12 months. Eight patients with a follow-up of less than 12 months were excluded from the study. In 136 patients (66.3%) the tumor did not grow. Forty-seven patients (23.9%) showed some evidence of slow growth. Eight of 197 patients (4%) had rapid growth and 6 patients (3%) had radiological evidence of tumor regression. Fifteen patients came to surgery. Five of these showed rapid growth, four developed ataxia in whom tumor growth was slow, three had ataxia without tumor growth, two patients developed brainstem compression, and one patient elected to proceed to surgery, although there were no tumor growth or symptoms. CONCLUSION: Few patients with small tumors will come to surgery in the short term. Perhaps the majority of patients with such small tumors will not need surgery. Long-term follow-up studies of 20 years or more are required to be come more confident about the natural history of these tumors. This study continues. PMID- 17077874 TI - Direct Facial-to-Hypoglossal Neurorrhaphy with Parotid Release. AB - OBJECTIVE: Facial nerve paralysis or compromise can be caused by lesions of the temporal bone and cerebellopontine angle and their treatment. When the facial nerve is transected or severely compromised and primary end-to-end repair is not possible, hypoglossal-facial nerve anastomosis remains the most popular method for accomplishing three main goals: restoring facial tone, restoring facial symmetry, and facilitating return of voluntary facial movement. Our objectives are to evaluate the surgical feasibility and long-term outcomes of our technique of direct facial-to-hypoglossal neurorrhaphy with a parotid-release maneuver. DESIGN: Prospective cohort. SETTING: Academic tertiary care referral center. PATIENTS: Ten patients with facial paralysis from proximal nerve injury underwent the facial-hypoglossal neurorrhaphy with a parotid-release maneuver. MAIN OUTCOME MEASURES: The Repaired Facial Nerve Recovery Scale, questionnaires, and photographs. RESULTS: Facial-hypoglossal neurorrhaphy with parotid release was technically feasible in all cases, and anastomosis was performed distal to the origin of the ansa hypoglossi. All patients had good return of facial nerve function. Nine patients had scores of C or better, indicating strong eyelid and oral sphincter closure and mass motion. There was no hemilingual atrophy and no subjective tongue dysfunction. CONCLUSIONS: The parotid-release maneuver mobilizes additional length to the facial nerve, facilitating a tensionless communication distal to the ansa hypoglossi. The technique is a viable option for facial reanimation, and our patients achieved good clinical outcomes with continual improvement. PMID- 17077875 TI - Early recurrence of an intracranial epidermoid cyst due to low-grade infection: case report. AB - Intracranial epidermoid cysts are slow-growing lesions, which may recur after incomplete resection, but do so over many years. The authors present the case of an epidermoid that repeatedly recurred over a short period, which was discovered to be secondary to a low-grade bacterial infection. A 30-year-old woman presented with signs and symptoms related to brainstem and cranial nerve compromise from a large cerebellopontine angle epidermoid cyst. She underwent a subtotal excision of a macroscopically and histologically typical epidermoid. The lesion recurred within 1 year of surgery, but with atypical radiological appearances. No systemic or local evidence of infection was found during a second resection. A further large recurrence with the same atypical features occurred after another 6 months, necessitating a third surgical resection. On this occasion, the atypical cyst contents grew coagulase-negative Staphylococcus after prolonged culture in an enrichment medium. Following a course of appropriate antibiotics, the residual cyst contents completely resolved. This case highlights an unusual complication of an epidermoid cyst. Early recurrence of an epidermoid, even in the absence of overt evidence of infection, should produce a high index of suspicion that there may be a low-grade infective cause. Surgical excision coupled with antibiotics resulted in a symptomatic and radiological recovery. PMID- 17077876 TI - Midfacial Degloving: The Best Alternative for Treatment of Trans-sphenoidal Meningocele of the Pterygopalatine Fossa. AB - Trans-sphenoid anterobasal temporal lobe meningoceles are rare and can be associated with temporal lobe epilepsy or recurrent meningitis. Surgical treatment is described via complicated infratemporal or intracranial approaches with high morbidity. A 32-year-old man presented with an 18-year history of two types of seizures and confirmed epileptic activity in electroencephalogram. A trans-sphenoid meningocele in the left pterygopalatine fossa was found on computed tomography and magnetic resonance imaging. A midfacial degloving was chosen instead of a large neurosurgical approach to remove it and to repair the dura defect with lyophilized dura, collagen, and abdominal fat. The postoperative course was uneventful without visible scars and the patient remains free of seizures without antiepileptic medication 3 years after surgery. Although midfacial degloving is not described yet as a treatment of trans-sphenoid meningoceles, this approach proved to be fast, safe, effective, and reliable and should be considered as the only alternative to large external approaches. PMID- 17077877 TI - Australian guidelines for the performance of the sweat test for the diagnosis of cystic fibrosis: report from the AACB Sweat Testing Working Party. PMID- 17077878 TI - Computerised order entry systems and pathology services--a synthesis of the evidence. AB - Computerised Physician Order Entry (CPOE) systems have been promoted in Australia and internationally for their potential to improve the quality of care. The existing research of the effect of CPOE on pathology laboratories has been variable, pointing to the potential to increase efficiency and effectiveness and contribute to enhancing the quality of patient care on the one hand, while leading to significant disruptions in work organisation with a negative impact on departmental relations on the other hand. In this paper we provide an overview of the research evidence about the impact of CPOE on four areas associated with pathology services; a) efficiency of the ordering process, e.g. test turnaround times, b) effectiveness as measured by test ordering volumes and test order appropriateness, c) quality of care, particularly its effects on patient care and d) work organisation patterns, which can be severely disrupted by CPOE. We discuss the possible ramifications of CPOE and offer three broad, but important recommendations for pathology laboratories, based on our own research experience investigating CPOE implementations over three years. Firstly, pathology laboratories need to be active participants in planning the implementation of CPOE. Secondly, the importance of building a firm organisational foundation for the introduction of the new system that includes openness and responsiveness to feedback. And thirdly, the implementation process needs to be underpinned by a strong commitment to a multi-method evaluation at every stage of the process to be able to measure the impact of the system on work practices and outcomes. PMID- 17077879 TI - Communication systems in healthcare. AB - The care of patients now almost inevitably seems to involve many different individuals, all needing to share patient information and discuss their management. As a consequence there is increasing interest in, and use of, information and communication technologies to support health services. Yet, while there is significant discussion of, and investment in, information technologies, communication systems receive much less attention and the clinical adoption of even simpler services like voice-mail or electronic mail is still not commonplace in many health services. There remain enormous gaps in our broad understanding of the role of communication services in health care delivery. Laboratory medicine is perhaps even more poorly studied than many other areas, such as the interface between primary care and hospital services. Given this lack of specific information about laboratory communication services, this paper will step back and generally review the components of a communication system, including the basic concepts of a communication channel, service, device and interaction mode. The review will then try and summarise some of what is known about specific communication problems that arise across health services in the main, including the community and hospital service delivery. PMID- 17077881 TI - Recent advances in tailoring the aggregation of heavier alkaline earth metal halides, alkoxides and aryloxides from non-aqueous solvents. AB - This overview on one of the subjects treated in our group deals with the synthesis and study of low-dimensional polymer and molecular solid state structures formed with alkaline earth metal ions in non-aqueous solvents. We have chosen several synthetic approaches in order to obtain such compounds. The first concept deals with the "cutting out" of structural fragments from a solid state structure of a binary compound, which will be explained with reference to BaI2. Depending on the size and concentration of oxygen donor ligands, used as chemical scissors on BaI2, three-, two-, one- and zero-dimensional derived adducts of BaI2 are obtained, comparable to a structural genealogy tree for BaI2. A second part deals with the supramolecular approach for the synthesis of low dimensional polymeric compounds based on alkaline earth metal iodides, obtained by the combination of metal ion coordination with hydrogen bonding between the cationic complexes and their anions. Certain circumstances allow rules to be established for the prediction of the dimensionality of a given compound, contributing to the fundamental problem of structure prediction in crystal engineering. A third section describes a synthetic approach for generating pure alkaline earth metal cage compounds as well as alkali and alkaline earth mixed metal clusters. A first step deals with different molecular solvated alkaline earth metal iodides which are investigated as a function of the ligand size in non-aqueous solvents. These are then reacted with some alkali metal compound in order to partially or totally eliminate alkali iodide and to form the targeted clusters. These unique structures of ligand stabilized metal halide, hydroxide and/or alkoxide and aryloxide aggregates are of interest as potential precursors for oxide materials and as catalysts. Approaches to two synthetic methods of the latter, sol-gel and (MO)CVD (metal-organic chemical vapour deposition), are investigated with some of our compounds. (D. C. Bradley, Chem. Rev., 1989, 89, 1317-22; K. G. Caulton and L. G. Hubert-Pfalzgraf, Chem. Rev., 1990, 90, 969-95; T. P. Hanusa, Coord. Chem. Rev., 2000, 210, 329-367; T. P. Hanusa, in Comprehensive Coordination Chemistry II, ed. J. A. McCleverty and T. J. Meyer, Elsevier, Amsterdam, 2004, vol. 3, 1 92). Finally, the physical properties of some of our compounds are described qualitatively in order to show the wide spectrum of possibilities and potential applications for chemistry in this field. PMID- 17077882 TI - Synthesis, structure and magnetic properties of the first Mn(II)-Gd(III) heteronuclear complex. AB - The synthesis and crystal structure of a novel pentanuclear Mn(II)-Gd(III) complex is reported; the magnetic measurement conclusively demonstrated anti ferromagnetic behaviour between Mn(II) and Gd(III) for this complex. PMID- 17077880 TI - Clinical proteomics: present and future prospects. AB - Advances in proteomics technology offer great promise in the understanding and treatment of the molecular basis of disease. The past decade of proteomics research, the study of dynamic protein expression, post-translational modifications, cellular and sub-cellular protein distribution, and protein protein interactions, has culminated in the identification of many disease related biomarkers and potential new drug targets. While proteomics remains the tool of choice for discovery research, new innovations in proteomic technology now offer the potential for proteomic profiling to become standard practice in the clinical laboratory. Indeed, protein profiles can serve as powerful diagnostic markers, and can predict treatment outcome in many diseases, in particular cancer. A number of technical obstacles remain before routine proteomic analysis can be achieved in the clinic; however the standardisation of methodologies and dissemination of proteomic data into publicly available databases is starting to overcome these hurdles. At present the most promising application for proteomics is in the screening of specific subsets of protein biomarkers for certain diseases, rather than large scale full protein profiling. Armed with these technologies the impending era of individualised patient tailored therapy is imminent. This review summarises the advances in proteomics that has propelled us to this exciting age of clinical proteomics, and highlights the future work that is required for this to become a reality. PMID- 17077883 TI - Rational ligand design for metal ion recognition. Synthesis of a N-benzylated N2S3-donor macrocycle for enhanced silver(I) discrimination. AB - Four previously documented ligand design strategies for achieving Ag(I) discrimination have been applied to the design of a new N-benzylated N2S3-donor macrocycle; the latter shows high selectivity for Ag(I) over Co(II), Ni(II), Cu(II), Zn(II), Cd(II) and Pb(II) in log K and bulk membrane transport studies. PMID- 17077884 TI - Reactivity of a palladium fluoro complex towards silanes and Bu3SnCH=CH2: Catalytic derivatisation of pentafluoropyridine based on carbon-fluorine bond activation reactions. AB - The chloro and azido complexes trans-[PdCl(4-C5NF4)(PiPr3)2] (3) and trans [Pd(N3)(4-C5NF4)(PiPr3)2] (4) can be prepared by reaction of [PdF(4 C5NF4)(PiPr3)2] (2) with Et3SiCl or MeSiN3, respectively. In contrast, reactions of 2 with Ph3SiH or Me2FSiSiFMe2 give the products of reductive elimination 2,3,5,6-tetrafluoropyridine (5) or 4-(fluorodimethylsilyl)tetrafluoropyridine (6) as well as [Pd(PiPr3)2] (1). In a catalytic experiment, pentafluoropyridine can be converted with Ph3SiH into 5 in 62% yield, when 10% of 2 is employed as catalyst. Treatment of trans-[PdF(4-C5NF4)(PiPr3)2] (2) with Bu3SnCH=CH2 in THF at 50 degrees C results in the formation of [Pd(PiPr3)2] (1) and 4 vinyltetrafluoropyridine (7). Complex 2 is also active as a catalyst towards a Stille cross-coupling reaction of pentafluoropyridine with Bu3SnCH=CH2 to give 4 vinyltetrafluoropyridine (7) with a TON of 6. The molecular structure of the complex 3 has been determined by X-ray crystallography. PMID- 17077885 TI - Experimental and computational evidence of the intermolecular motifs in the crystal packing of luminescent pentacoordinated gallium(III) complexes. AB - This paper reports the synthesis, characterization, photophysical and structural properties of the homologous series of good emitting pentacoordinated GaQ'2L complexes 1-3, where Q' is 2-methyl-quinolin-8-olate and L is a phenolate substituted in para position with respect to the oxygen donor atom. A combined approach between the experimental structural analysis (i.e. the molecular fragments involved in intermolecular pi-pi interactions) and the computational study (i.e. the nature of the molecular orbitals residing therein) is discussed in order to compare the charge transport aptitude of complexes 1-3, in relation to the facing of their LUMO/LUMO, HOMO/HOMO and HOMO/LUMO arising from the molecular packing. The different phenolate ligands significantly change the packing characteristics of 1-3, with indirect effects on the electron hopping kinetics responsible for conduction in amorphous thin films. The observed preference for pyridyl-pyridyl stacking proves a faster electron conduction in comparison to hole conduction in the three complexes studied. PMID- 17077886 TI - Sulfate selective anion recognition by a novel tetra-imidazolium zinc metalloporphyrin receptor. AB - Imidazolium groups have been successfully incorporated into the structure of a "picket fence" porphyrin molecule to produce a novel tetra-imidazolium zinc metalloporphyrin anion receptor. UV/visible spectroscopic studies reveal that this receptor is selective for sulfate anions, capable of strongly complexing sulfate in competitive water-DMSO (5 : 95) solvent mixtures. Cyclic and square wave voltammetric studies demonstrate the receptor's ability to sense a variety of anions electrochemically. PMID- 17077887 TI - Synthesis and characterizations of cyclic octanuclear mixed-valence vanadium(IV,V) clusters with polyoxometalate counterions. AB - Hydrothermal reaction of Na2WO4, VOSO4, 2,2'-bpy and H3PO4 has afforded in high yield the compound [V(IV)2V(V)6O14(bpy)8(PO4)2][PW11V(V)O40](bpy).12H2O (1). Compound 1 contains a novel octanuclear mixed valence V(IV,V) cluster, [V(IV)2V(V)6O14(bpy)8(PO4)2]4+, with [PW11V(V)O40]4- as counterion. In the vanadium cluster, four V(V) centers are localized and the remaining two V(IV) and two V(V) ions are disordered over four crystallographically equivalent positions. The isostructural compound [V(IV)2V(V)6O14(bpy)8(PO4)2][PMo11V(V)O40](bpy).3H2O (2) has also been synthesized. Thermodiffractometry experiments indicate that 2 is stable up to 360 degrees C. Redox activities for both the vanadium and molybdenum centers have been observed by solid-state electrochemical measurements performed on mechanically attached microparticles of 2. Magnetic measurements performed on have shown the occurrence of weak ferromagnetic interactions between the V(IV) centres (J = +0.34 cm(-1), H(ex) = -JS1 x S2), and combined with DFT calculations, have allowed to propose a localization of the two V(IV) centers on two of the four equivalent crystallographic sites. Finally high field electron paramagnetic resonance has evidenced the magnetic axial anisotropy of the paramagnetic centers (g(x) = g(y) = 1.975(3); g(z) = 1.939(4)). PMID- 17077888 TI - Synthesis and characterization of a novel paramagnetic macromolecular complex [Gd(TTDASQ-protamine)]. AB - Adenocarcinomas in rats and humans frequently contain perivascular, degranulating mast cells that release heparin. Protamine is a low-molecular weight, cationic polypeptide that binds to heparin and neutralizes its anticoagulant properties. A novel magnetic resonance imaging (MRI) contrast agent containing protamine was synthesized. TTDASQ, the derivative of TTDA (3,6,10-tri(carboxymethyl)-3,6,10 triazadodecanedioic acid), was also synthesized and the kinetic stability of [Gd(TTDASQ)]- chelate containing phosphate buffer and ZnCl2 to measure the relaxation rate (R1) at 20 MHz was studied by transmetallation with Zn(II). The water-exchange rate (k(ex)298) of [Gd(TTDASQ)]- is 6.4 x 10(6) s(-1) at 25.0 +/- 0.1 degrees C which was obtained from the reduced 17O relaxation rates (1/T(1r) and 1/T(2r)) and chemical shift (omega(r)) of H(2)17O, and it is compared with that previously reported for the other gadolinium(III) complex, [Gd(DO3ASQ)]. The binding affinity assay showed that the (TTDASQ)3-pro19 has higher activity toward heparin. On the other hand, the effect of heparin on the relaxivity of the [Gd(TTDASQ)3-pro19] conjugate shows the binding strength (K(A)) is 7669 dm3 mol( 1) at pH 7.4 and the relaxivity (r(b)1) of the [Gd(TTDASQ)3-pro19]-heparin adduct is 30.9 dm3 mmol(-1) s(-1). PMID- 17077889 TI - Synthesis, structure and catalase-like activity of dimanganese(III) complexes of 1,5-bis(X-salicylidenamino)pentan-3-ol (X = 3- and 5-methyl). Influence of phenyl ring substituents on catalytic activity. AB - The diMn(III) complexes [Mn2(5-Me-salpentO)(mu-MeO)(mu-AcO)(H2O)Br] (1) and [Mn2(3-Me-salpentO)(mu-MeO)(mu-AcO)(MeOH)2]Br (2), where salpentOH = 1,5 bis(salicylidenamino)pentan-3-ol, were synthesised and structurally characterized. The two complexes include a bis(micro-alkoxo)(micro-acetato) triply-bridged diMn(III) core with an Mn...Mn separation of 2.93-2.94 A, the structure of which is retained upon dissolution. Complexes 1 and 2 show catalytic activity toward disproportionation of H2O2, with first-order dependence on the catalyst, and saturation kinetics on [H2O2], in methanol and DMF. In DMF, the two complexes are able to disproportionate at least 1500 eq. of H2O2 without significant decomposition, while in methanol, they rapidly lose activity with formation of a non-coupled Mn(II) species. Electrospray ionisation mass spectrometry, EPR and UV/vis spectroscopy used to monitor the reaction suggest that the major active form of the catalyst occurs in the Mn2(III) oxidation state during cycling. The correlation between log(k(cat)) and the redox potentials of 1, 2 and analogous complexes of other X-salpentOH derivatives indicates that, in this series, the oxidation of the catalyst is probably the rate-limiting step in the catalytic cycle. It is also noted that formation of the catalyst-peroxide adduct is more sensitive to steric effects in DMF than in methanol. Overall, kinetics and spectroscopic studies of H2O2 dismutation by these complexes converge at a catalytic cycle that involves the Mn2(III) and Mn2(IV) oxidation states. PMID- 17077890 TI - Insertion reaction of carbon dioxide into Sn-OR bond. Synthesis, structure and DFT calculations of di- and tetranuclear isopropylcarbonato tin(IV) complexes. AB - The reaction of carbon dioxide with the stannane nBu2Sn(OiPr)2 and distannoxane [nBu2(iPrO)Sn]2O leads to the selective insertion into one Sn-OiPr bond generating the corresponding nBu2Sn(OiPr)(OCO2(i)Pr) and nBu2(iPrO)SnOSn(OCO2(i)Pr)nBu2 species. Both compounds are characterised by multinuclear NMR, FT-IR and single-crystal X-ray crystallography. In the solid state, they adopt a dimeric arrangement with bridging isopropoxy and terminal isopropylcarbonato ligands. The X-ray crystal structure of the dinuclear stannane shows that the Sn2O2 ring and the two Sn-OCO2C fragments are nearby coplanar. The same holds for the ladder-type tetranuclear distannoxane. The dimeric structures are also evidenced by solution NMR in non-coordinating solvents. Interestingly, the assignment of the exo and endo tin resonances of the dimeric distannoxane is unambiguous using a labeled 13CO2 experiment. The stability of the dimeric association has been probed in the stannane series on the basis of DFT calculations. PMID- 17077891 TI - Structure, equilibrium and ligand exchange dynamics in the binary and ternary dioxouranium(VI)-ethylenediamine-N,N'-diacetic acid-fluoride system: A potentiometric, NMR and X-ray crystallographic study. AB - The structure, thermodynamics and kinetics of the binary and ternary uranium(VI) ethylenediamine-N,N'-diacetate (in the following denoted EDDA) fluoride systems have been studied using potentiometry, 1H, 19F NMR spectroscopy and X-ray diffraction. The UO2(2+)-EDDA system could be studied up to -log[H3O+] = 3.4 where the formation of two binary complexes UO2(EDDA)(aq) and UO2(H3EDDA)3+ were identified, with equilibrium constants logbeta(UO2EDDA) = 11.63 +/- 0.02 and logbeta(UO2H3EDDA3+) = 1.77 +/- 0.04, respectively. In the ternary system the complexes UO2(EDDA)F-, UO2(EDDA)(OH)- and (UO2)2(mu-OH)2(HEDDA)2F2(aq) were identified; the latter through 19F NMR. 1H NMR spectra indicate that the EDDA ligand is chelate bonded in UO2(EDDA)(aq), UO2(EDDA)F- and UO2(EDDA)(OH)- while only one carboxylate group is coordinated in UO2(H3EDDA)3+. The rate and mechanism of the fluoride exchange between UO2(EDDA)F- and free fluoride was studied by 19F NMR spectroscopy. Three reactions contribute to the exchange; (i) site exchange between UO2(EDDA)F- and free fluoride without any net chemical exchange, (ii) replacement of the coordinated fluoride with OH- and (iii) the self dissociation of the coordinated fluoride forming UO2(EDDA)(aq); these reactions seem to follow associative mechanisms. (1)H NMR spectra show that the exchange between the free and chelate bonded EDDA is slow and consists of several steps, protonation/deprotonation and chelate ring opening/ring closure, the mechanism cannot be elucidated from the available data. The structure (UO2)2(EDDA)2(mu-H2EDDA) was determined by single crystal X-ray diffraction and contains two UO2(EDDA) units with tetracoordinated EDDA linked by H2EDDA in the "zwitterion" form, coordinated through a single carboxylate oxygen from each end to the two uranium atoms. The geometry of the complexes indicates that there is no geometric constraint for an associative ligand substitution mechanism. PMID- 17077892 TI - Thermodynamic, kinetic and solid-state study of divalent metal complexes of 1,4,8,11-tetraazacyclotetradecane (cyclam) bearing two trans (1,8 )methylphosphonic acid pendant arms. AB - Divalent metal complexes of macrocyclic ligand 1,4,8,11-tetraazacyclotetradecane 1,8-bis(methylphosphonic acid)) (1,8-H4te2p, H4L) were investigated in solution and in the solid state. The majority of transition-metal ions form thermodynamically very stable complexes as a consequence of high affinity for the nitrogen atoms of the ring. On the other hand, complexes with Mn2+, Pb2+ and alkaline earth ions interacting mainly with phosphonate oxygen atoms are much weaker than those of transition-metal ions and are formed only at higher pH. The same tendency is seen in the solid state. Zinc(II) ion in the octahedral trans O,O-[Zn(H2L)] complex is fully encapsulated within the macrocycle (N4O2 coordination mode with protonated phosphonate oxygen atoms). The polymeric {[Pb(H2L)(H2O)2].6H2O}n complex has double-protonated secondary amino groups and the central atom is bound only to the phosphonate oxygen atoms. The phosphonate moieties bridge lead atoms creating a 3D-polymeric network. The [{(H2O)5Mn}2(micro-H2L)](H2L).21H2O complex contains two pentaaquamanganese(II) moieties bridged by a ligand molecule protonated on two nitrogen atoms. In the complex cation, oxygen atoms of the phosphonate groups on the opposite sites of the ring occupy one coordination site of each metal ion. The second ligand molecule is diprotonated and balances the positive charge of the complex cation. Complexation of zinc(II) and cadmium(II) by the ligand shows large differences in reactivity of differently protonated ligand species similarly to other cyclam like complexes. Acid-assisted dissociations of metal(II) complexes occur predominantly through triprotonated species [M(H3L)]+ and take place at pH < 5 (Zn2+) and pH < 6 (Cd2+). PMID- 17077893 TI - Photo-assisted chlorination of cycloalkanes with iron chloride heterogenized with Amberlite. AB - Photoexcitation under mild temperature and pressure conditions of FeCl4- heterogenized with Amberlite causes the conversion of several cycloalkanes to the corresponding monochlorinated products with selectivity higher than 95%. PMID- 17077894 TI - Synthesis of estradiol-pheophorbide a conjugates: evidence of nuclear targeting, DNA damage and improved photodynamic activity in human breast cancer and vascular endothelial cells. AB - The synthesis, physico-chemical properties, cellular localization and photocytotoxicity of estradiol-pheophorbide a conjugates in estrogen-dependent cancer and vascular endothelial cells are described with the aim of increasing the photodynamic activity by targeting the nucleus of both tumor and blood vessel cells. PMID- 17077895 TI - Photochemical synthesis of benzoyl spiro[2.2]pentanes. AB - In the present study, we describe the photochemical behaviour of 2-mesyloxy phenyl ketones 8 and 12 bearing a cyclopropane moiety in the side-chain. Irradiation of 8 and 12 leads to the corresponding benzoyl spiro[2.2]pentanes as a consequence of an initial gamma-H-shift, subsequent elimination of MsOH (accompanied by a spin-center shift) and cyclization of the resulting 1,3 diradicals. In contrast, a corresponding phenyl ketone without a mesyloxy group in the 2-position, and thus a potential reactant of the "classical" Norrish-Yang reaction, shows no photochemical reaction. By means of quantum chemical calculations we discovered that in the presence of a mesyloxy group the activation barrier for the photochemical gamma-H-shift is substantially decreased. Furthermore, a photoinduced skeletal rearrangement of benzoyl spiro[2.2]pentane to 2-cyclobutylidene-acetophenone could be observed. Compared to the common methods used to synthesize spiro[2.2]pentanes, the photochemical preparation of benzoyl spiro[2.2]pentane presented herein is the first example where a bond between the spiro atom and an adjacent atom is formed. PMID- 17077896 TI - Intravenous repeated-dose toxicity study of ZnPcS2P2-based-photodynamic therapy in Wistar rats. AB - The purpose of the current study was to investigate the potential repeated-dose toxicity of ZnPcP2S2-based photodynamic therapy (ZnPc-PDT) in Wistar rats. The animals were administered ZnPcS2P2 intravenously ten times successively every 4 d and irradiated with a 670 nm laser light for 6 min at subsequent 48 h and 72 h. At the end of the treatment period, 10 rats/sex/group were sacrificed, while 5 rats/sex/group were sacrificed after a two-week recovery period. During the test period, clinical signs, mortality, body weights, food and water consumption, ophthalmoscopy, hematology, serum biochemistry, urinalysis, organ weights, gross findings and histopathology were examined. The association between the increased liver weight and hepatic spotty and lytic necrosis seen in high dose females corroborates the conclusion that high dose ZnPc-PDT could induce hepatic injury in Wistar rats and they are probably related to the abnormality of certain biochemical parameters of females in the high dose group. Furthermore, microscopic examination for the ZnPc-PDT groups shows the presence of some Kelly and khaki granules in Kupffer cells and endothelia of the livers, epithelia of the renal tubules, marginal sinus and medulla of the spleens, alveolar walls of the lungs, reticular cells and macrophages of the mesenteric lymph nodes, testicular Leydig cells, epididymal epithelial cells, endometrial stromal cells, and interstitial cells and corpora lutea of the ovaries from all or most of the animals. There were no adverse effects on mortality, clinical signs, food and water consumption, ophthalmoscopy, uranalysis, hematology, serum biochemistry, body weights and necropsy findings in control, low and mid dose groups. Based on these results, it was concluded that the intravenous repeated-dose of ZnPcP2S2 PDT induced the abnormalities of liver weights, hepatic biochemistry and histopathology, and pigmentation in the several important organs in Wistar rats at 4 mg kg(-1) d(-1). The target organ was determined to be liver (and spleen perhaps), but this was not so obvious in males. The no-observed-adverse-effect level (NOAEL) was considered to be 1.0 mg kg(-1) for both sexes. PMID- 17077897 TI - Spectroscopic characterization and photobleaching kinetics of hypericin-N-methyl pyrrolidone formulations. AB - Hypericin (HY) is a promising photosensitizer in photodynamic therapy (PDT). It was recently reported that appropriate use of N-methyl pyrrolidone (NMP) enhanced in vivo PDT efficacy of HY and enhanced in vivo delivery of HY. This present study further investigates the use of NMP and other known non-toxic pharmaceutical additives, polyvinylpyrrolidone (PVP, K29/32) and copolyvidonum (S630), for formulating HY to enhance its delivery with photodynamic activity as a goal in mind. Hence, the first objective of this study was to characterize the solubilization of HY by NMP, K29/32 and S630. Thermodynamic considerations were used to explain the solvation process. Photobleaching is another important property of photosensitizers. There is no report on the photostability of HY in pharmaceutical formulations used for PDT. Therefore, the second objective of this study was to investigate the photobleaching of HY in these formulations. The fluorescence of HY was found to increase significantly in higher concentrations of NMP or when 5% of polymer was co-mixed with 5% of NMP solution. The photobleaching of HY in these formulations followed first-order kinetics. The loss of fluorescence paralleled to the loss of absorption of HY. The formulation of HY with 40% NMP was found to be the most stable. PMID- 17077898 TI - Two-photon absorption of copper tetrasulfophthalocyanine induces phototoxicity towards Jurkat cells in vitro. AB - The feasibility to induce oxygen-independent tumour cell kill by two-photon excitation of copper tetrasulfophthalocyanine (CuPcS4) was studied in Jurkat cells in vitro. Following incubation with CuPcS4 cells were transferred to a closed cuvette and irradiated with 532 nm pulsed-laser or 680 nm continuous-laser light to evaluate the effect of either two- or one-photon excitation, respectively. Cell survival was measured using MTT and Trypan Blue exclusion tests. Cell viability decreased 10-20% following two-photon excitation while one photon illumination did not affect cell survival. These data confirm that two photon excitation of CuPcS4 to the upper excited triplet state results in the formation of toxic species suggesting its potential use as a sensitizer for the photodynamic treatment of poorly oxygenated tumours. PMID- 17077899 TI - The potential application of chlorin e6-polyvinylpyrrolidone formulation in photodynamic therapy. AB - Much research has been focused on developing effective drug delivery systems for the preparation of chlorins as potential photosensitizers for PDT. This report describes the evaluation of a new water-soluble formulation of chlorin e6 consisting of a complex of trisodium salt chlorin e6 and polyvinylpyrrolidone (Ce6-PVP) for application in photodynamic therapy (PDT) with 2 specific aims: (i) to investigate its fluorescence kinetics in skin, normal and tumor tissue after intravenous administration, and (ii) to investigate its PDT efficacy. Our results demonstrate that this new formulation possesses photosensitizing properties with rapid accumulation in tumor tissue observed within 1 h after intravenous administration. Although high selectivity in tumor tissue was found between the period of 3 and 6 h, the efficacy of Ce6-PVP mediated PDT was best at 1 h drug light interval. It is suggested that, the extent of tumor necrosis post PDT is dependent on the plasma concentration of Ce6-PVP, implying a vascular mediated cell death mechanism. A faster clearance rate of Ce6-PVP from the skin of nude mice was observed compared to Ce6. The new formulation of Ce6-PVP seems to show promise as an effective therapeutic agent. PMID- 17077900 TI - Modulation of dual fluorescence in a 3-hydroxyquinolone dye by perturbation of its intramolecular proton transfer with solvent polarity and basicity. AB - A representative of a new class of dyes with dual fluorescence due to an excited state intramolecular proton transfer (ESIPT) reaction, namely 1-methyl-2-(4 methoxy)phenyl-3-hydroxy-4(1H)-quinolone (QMOM), has been studied in a series of solvents covering a large range of polarity and basicity. A linear dependence of the logarithm of its two bands intensity ratio, log(I(N*)/I(T*)), upon the solvent polarity expressed as a function of the dielectric constant, (epsilon- 1)/(2epsilon + 1), is observed for a series of protic solvents. A linear dependence for log(I(N*)/I(T*)) is also found in aprotic solvents after taking into account the solvent basicity. In contrast, the positions of the absorption and the two emission bands of QMOM do not noticeably depend on the solvent polarity and basicity, indicating relatively small changes in the transition moment of QMOM upon excitation and emission. Time-resolved experiments in acetonitrile, ethyl acetate and dimethylformamide suggest an irreversible ESIPT reaction for this dye. According to the time-resolved data, an increase of solvent basicity results in a dramatic decrease of the ESIPT rate constant, probably due to the disruption of the intramolecular H-bond of the dye by the basic solvent. Due to this new sensor property, 3-hydroxyquinolones are promising candidates for the development of a new generation of environment-sensitive fluorescence dyes for probing interactions of biomolecules. PMID- 17077901 TI - Low ionic strength reduces cytosine photoreactivity in UVC-irradiated isolated DNA. AB - Exposure of DNA to far-UV radiation leads to the formation of several types of dimeric lesions, including cyclobutane dimers and (6-4) photoproducts. In order to gain insights into the main parameters driving DNA photochemistry, the effect of ionic strength on the yield of formation of these photoproducts was investigated in UVC-irradiated samples of isolated genomic DNA. The main consequence of lowering the ionic strength was a decrease in the UVC-induced formation of thymine-cytosine and cytosine-cytosine photoproducts. The reactivity of thymine-thymine and cytosine-thymine doublets was hardly affected. Evidence was obtained against a major role played by duplex denaturation in these observations. A more likely explanation is a change in the DNA structure as the result of a larger extent of protonation at low counter-ions concentration. PMID- 17077902 TI - Photoinduced oxygen uptake for 9,10-anthraquinone in air-saturated aqueous acetonitrile in the presence of formate, alcohols, ascorbic acid or amines. AB - The photolysis of 9,10-anthraquinone (AQ), 2-methyl- and 2,3-dimethyl-AQ was studied in air-saturated acetonitrile-water in the presence of various donors: formate, ascorbic acid, alcohols, e.g. 2-propanol or methanol, and amines, e.g. ethylenediaminetetraacetate (EDTA). The photoreaction is initiated by H-atom or electron transfer from the donor to the AQ triplet state. The conversion of oxygen into hydrogen peroxide occurs via the superoxide radical and its conjugate acid. The quantum yield of oxygen uptake (Phi(-O2)) increases with increasing donor concentration. Phi(-O2) = 0.3-0.6 in the presence of 1 M 2-propanol and 3 10 mM ascorbic acid or EDTA. The properties of the quinone and donor radicals involved and the pH and concentration dependences of Phi(-O2) are described. PMID- 17077903 TI - Photochemistry of 2,6-diisopropylphenol (propofol). AB - The photochemistry of the anaesthetic agent propofol (PPF) was investigated in three different solvents of quite different polarity (cyclohexane, methanol and phosphate buffer pH 7) by means of nanosecond laser flash photolysis and absorption spectroscopy. GC-MS spectrometry measurements of PPF in cyclohexane have revealed the formation of two major products upon low intensity UV continuous irradiation of PPF in aerated solution: the diphenol derivative of PPF and 2,6-diisopropyl-p-benzoquinone (PPFQ). Only the diphenol compound was obtained in anaerobic solution. PPF phenoxyl radical (PPF ) generation has been assigned as the original step leading to the formation of both the diphenol compound and PPFQ in cyclohexane as revealed by laser flash photolysis at 266 nm and by electron paramagnetic resonance spectroscopy as well. Investigation of PPF by nanosecond flash photolysis at 266 nm in the other solvents revealed the occurrence of different photochemical processes depending on the nature and the polarity of the solvent. A reaction scheme is proposed in order to discuss the mechanism of reaction of PPF in all media. PMID- 17077904 TI - Pyrene-p-tert-butylcalixarenes inclusion complexes formation: a surface photochemistry study. AB - Diffuse reflectance and luminescence techniques were used to study the photophysics and photochemistry of pyrene within p-tert-butylcalix[n]arenes with n = 4, 6, and 8, and to study their ability to form inclusion complexes in heterogeneous media. Evidences for inclusion complex formation were found for the three hosts under study. Ground state diffuse reflectance results have shown the formation of ground state dimers of pyrene inside the cavity of calix[6]arene and calix[8]arene, with this feature much more evident for calix[6]arene. For calix[4]arene, only a monomer fits inside the cavity and the presence of pyrene microcrystals outside the cavity was detected. A luminescence lifetime distribution analysis was performed, revealing the presence of prompt emissions from the pyrene microcrystals outside the cavity in the case of calix[4]arene and from the constrained dimers inside the cavities of calix[6]arene and calix[8]arene. Transient absorption results have shown the presence of pyrene radical cation and also of trapped electrons for the three hosts under study. The formation of the phenoxyl radical of the calixarene following the laser pulsed excitation of pyrene at 355 nm is increased for calix[6]arene and calix[8]arene. This feature is particularly relevant for calix[6]arene, suggesting a very favourable situation for the hydrogen atom abstraction to occur. The analysis of the degradation products revealed the presence of hydroxypyrene as a major photodegradation product for the three hosts. Dihydro-hydroxypyrene was also formed in the case of calix[6]arene and calix[8]arene. The formation of the calixarene's phenoxyl radical and subsequent hydrogen abstraction is consistent with the formation of dihydro-dihydroxypyrene. PMID- 17077905 TI - Nonlinear regression analysis of kinetics of the photocatalytic decolorization of an azo dye in aqueous TiO2 slurry. AB - The kinetics of decolorization of an anionic monoazo dye of acid class named C.I. Acid Red 27 (AR27) was investigated in the UV/TiO2 process with nonlinear regression analysis. The experimental results indicated that the kinetics of decolorization of AR27 in this process fit well by pseudo-first order kinetics. With nonlinear regression analysis a model was developed for pseudo-first order rate constant (k(ap,UV/TiO2)) as a function of operational parameters such as TiO2 dosage, initial concentration of AR27, concentration of O2 and UV-light intensity (I0) as following: k(ap,UV/TiO2) = 0.0025 [TiO2](0.65) [AR27]0(-0.96) [O2](0.16)I0. This rate expression can be used for predicting k(ap,UV/TiO2) at different conditions. PMID- 17077907 TI - Establishment and support of a children's hospital--the community as champion. PMID- 17077906 TI - Imaging gene expression in live transgenic mice after providing luciferin in drinking water. AB - Mice expressing the firefly luciferase gene luc under the control of various gene promoters are used to image long-term changes in tumor growth, infection, development, and circadian rhythms. This novel approach enables ongoing regulation of gene expression to be visualized through repeated imaging of luciferase bioluminescence. Typically, luciferin, the luciferase substrate, is injected into mice before they are anaesthetized for imaging. To avoid the effects of handling and stress from injection on expression of the transgene, oral luciferin delivery methods were tested as an alternative to current methods. For unobscured imaging, a transgenic mouse line containing luc controlled by the enhancer and promoter for the major immediate-early gene of human cytomegalovirus (CMV) was crossed with a hairless albino mouse stock (HRS/J), resulting in the Hr CMV line. Mice given food and water ad libitum readily drank 1-5 mM luciferin in water or apple juice and could be imaged repeatedly on subsequent days without any apparent adverse effects. Oral and injected luciferin produced similar patterns of luminescence in the body areas examined: abdomen, tail vertebrae, gonads, hind leg, foreleg and others, although the tail showed a slightly brighter relative luminescence after oral luciferin. These results show that luciferin is not appreciably degraded in the digestive tract and can be easily administered orally to avoid injection and any concomitant effects on behavior that could alter gene expression. PMID- 17077908 TI - The rise and fall of children's hospitals in South Africa. PMID- 17077909 TI - The history of a War Memorial Children's Hospital in Cape Town. PMID- 17077910 TI - Undernutrition, brain growth and intellectual development. PMID- 17077911 TI - Observations on the origin of congenital intestinal atresia. PMID- 17077912 TI - Appendicitis--pitfalls and medicolegal implications. PMID- 17077913 TI - The history of paediatric trauma care in Cape Town. PMID- 17077914 TI - Fifty years of paediatric anaesthesia--new approaches to an old technique. PMID- 17077915 TI - Childhood pneumonia--progress and challenges. AB - Remarkable progress has been made in the development of antimicrobial therapy, effective vaccines and pneumonia management guidelines in the past 50 years. However, pneumonia is currently the leading cause of death in children younger than 5 years in developing countries, accounting for approximately 20% of childhood deaths. This article reviews changes in the epidemiology, management and prevention of childhood pneumonia in developing countries, specifically in Africa and South Africa, and addresses future challenges. MAIN FINDINGS: The HIV epidemic has sharply increased the incidence, severity of, and mortality due to, childhood pneumonia. Bacterial infection remains a major cause of pneumonia mortality. Additional pathogens such as Pneumocystis jirovecii and Gram-negative bacteria are found in HIV-infected children, associated with a high mortality. Mycobacterium tuberculosis is an important cause of acute pneumonia in both HIV infected and uninfected children. Use of case management guidelines can substantially reduce neonatal, infant and under-5 mortality and pneumonia specific mortality. General preventive interventions including micronutrient supplementation with zinc and vitamin A, and immunisations can substantially reduce the burden of childhood pneumonia. Despite a lower efficacy in HIV infected children, vaccination protects against disease in a significant proportion of children. In South Africa, new advances over the past 50 years have included greater access to primary health care for children, the use of Integrated Management of Childhood Illness guidelines in primary care, development of guidelines for diagnosis and management of childhood pneumonia and adoption of an expanded immunisation programme that includes coverage for Haemophilus influenzae type b. The pneumococcal conjugate vaccine recently licensed in South Africa also has the potential to significantly reduce the burden of childhood pneumonia. Recent roll-out of the national antiretroviral programme can reduce the incidence and severity of HIV-associated pneumonia through the prevention of HIV infection, use of cotrimoxazole prophylaxis and treatment with antiretrovirals. CONCLUSION: Available, effective interventions for prevention and treatment of childhood pneumonia exist; the challenge is to achieve widespread implementation and high coverage rates in developing countries. Greater access to newer vaccines and to antiretroviral therapy and co trimoxazole prophylaxis in HIV-infected children is necessary to further reduce the burden of childhood pneumonia and the discrepancies in global child lung health. PMID- 17077916 TI - The Allergy Clinic: 50 years. PMID- 17077917 TI - Sydenham's chorea--clinical and therapeutic update 320 years down the line. PMID- 17077918 TI - Why are some South African children with Down syndrome not being offered cardiac surgery? AB - About 1 in 1,000 children has Down syndrome. Extra chromosomal material results in a myriad of potential problems for the affected individual. About 40% of Down syndrome children will have cardiac abnormalities, ranging from the simple arterial duct to the complex atrioventricular septal defect. Virtually all these defects are amenable to surgical correction and extended survival is possible. In South Africa many of these children do not undergo cardiac surgery. PMID- 17077919 TI - Is cardiac surgery warranted in children with Down syndrome? A case-controlled review. AB - OBJECTIVES: To compare children with Down syndrome and children without Down syndrome and investigate whether there is a significant difference in the burden that is placed on the health care system between these two groups only in respect of the repair of congenital heart disease at Red Cross War Memorial Children's Hospital, Cape Town, South Africa. DESIGN: This study is a retrospective case control review. SETTING: Red Cross War Memorial Children's Hospital, Cape Town, South Africa. SUBJECTS: The sample group of 50 Down syndrome children who had received cardiac surgery between January 1998 and June 2003 was compared with a control group of 50 nonsyndromic children who had received cardiac surgery during the same period. OUTCOME MEASURES: Sex and diagnoses (cardiac and noncardiac), number of days spent in hospital and in ICU, complication rates, re-operation rates, early mortality rates, planned further cardiac surgery. Costs of these outcomes were not quantified in exact monetary terms. RESULTS: There was no significant difference between the two groups in terms of the burden that was placed on the health care system. Similar complication rates, re-operation rates and early mortality rates were recorded for both groups. The Down syndrome group appeared to benefit more from cardiac surgery than the non-Down syndrome group. CONCLUSION: Denying cardiac surgery to children with Down syndrome does not improve the efficiency of resource allocation. It is therefore not reasonable to suggest that the problem of scarce resources can be ameliorated by discriminating against children with Down syndrome. PMID- 17077920 TI - Four decades of conjoined twins at Red Cross Children's Hospital--lessons learned. AB - Conjoined twins represent a rare but fascinating congenital condition, the aetiology of which remains obscure. Over the past four decades, the paediatric surgeons at Red Cross Children's Hospital have been involved in the management of 46 pairs of conjoined twins, of which 33 have been symmetrical and 12 asymmetrical. Seventeen symmetrical twins have undergone separation with 22 children (65%) surviving; all of the live asymmetrical twins survived separation. We describe the important features of this unique cohort, outline our approach to management and present the results of this approach. We consider some of the ethical and moral dilemmas we have confronted, and discuss the prenatal diagnosis, obstetric implications and postnatal care of these children, including the relevant investigations and anaesthetic and surgical management. Specific aspects related to the cardiovascular system, hepatobiliary and gastrointestinal tracts, urogenital tract, central nervous system and musculoskeletal system are highlighted. PMID- 17077921 TI - The utility of computed tomography for recent-onset partial seizures in childhood. AB - OBJECTIVES: To determine the diagnostic yield of computed tomography (CT) of the head in children presenting for the first time with partial seizures in a region with a high prevalence of tuberculosis and neurocysticercosis. DESIGN: Prospective cohort study. SETTING: The secondary-level ambulatory service of Red Cross Children's Hospital, Cape Town. SUBJECTS: Children aged 6 months-12 years with a first partial seizure. Outcome measures. Abnormal CT findings; clinically unsuspected abnormal CT findings. RESULTS: Of 118 enrolled children, CT findings were available for 94 (80%). Sixteen (33%) of 49 children scheduled to return later for an initial CT scan failed to do so. Thirty-two scans (34%) were reported normal, 45 (48%) showed single or multiple granulomas, and 17 (18%) showed other findings. All 8 children with persistent specific CT findings were suspected of having the condition before CT scan. Of 68 cases with prospectively recorded clinically expected CT findings, normal scans were expected in 2 cases (3%) and occurred in 33 cases (49%). CONCLUSIONS: Routine CT scan for children presenting with a first partial seizure in an area with a high prevalence of neurocysticercosis failed to identify findings other than neurocysticercosis that meaningfully altered clinical management. Assuming a 70% relative reduction of seizures with albendazole treatment for neurocysticercosis, routine CT scanning in the study population would require 11 scans and 5 courses of albendazole to prevent 1 child from having seizures, compared with no CT scans and 11 courses of albendazole with blanket use of albendazole. PMID- 17077922 TI - The impact of subspecialty services on health care delivery--a community health centre based study. AB - OBJECTIVES: The objective was to evaluate the role of a paediatric surgical consultant at a primary health care facility. DESIGN: Descriptive and prospective. SETTING: In the process of planning and implementation of the 2010 health plan of the Provincial Government of the Western Cape, a shift occurred in the delivery of health care to children from a provincially based hospital system to a municipally based primary health care system. To contribute towards enabling this process, the Department of Paediatric Surgery at Red Cross War Memorial Children's Hospital established a paediatric surgical day clinic at a local community health centre during 2001. SUBJECTS: Information was obtained from patient data sheets containing details of consultations at the sub-specialist surgical clinic at Michael Mapongwana Community Health Centre. RESULTS: Over a 58 month period 1 171 children were seen, of whom 655 were male and 427 female. Their ages ranged from 0 to 19 years, the largest group being under 1 year. Eighty per cent of patients were accompanied by their mothers. The correct diagnosis was established by the nurse practitioners in 71%. General paediatric surgical conditions predominated, followed by medical, dermatological, orthopaedic, trauma, otolaryngo-pharyngology, infectious diseases, ophthalmology, urology, neurosurgery, malignancy and maxillofacial conditions. The details are set out in the report. In total 597 patients were referred directly to an appropriate care facility and 574 patients could be managed entirely at the clinic level. CONCLUSIONS: This study demonstrated the significant public health problem of paediatric surgical disease. It emphasized the preventative and cost effective role of a surgical clinic at primary health care level. The clinic allowed for timely surgical intervention in 65% of surgical cases, thereby decreasing inappropriate tertiary referrals. We believe that bringing specialists into the community can only strengthen the 2010 health care plan. PMID- 17077923 TI - The evolving management of Burkitt's lymphoma at Red Cross Children's Hospital. AB - BACKGROUND: Treatment for Burkitt's lymphoma at Red Cross Children's Hospital has evolved from the use of aggressive surgery and less intensive chemotherapy to a conservative surgical approach with more intensive chemotherapy. METHODS: The study was a retrospective folder review of patients diagnosed with Burkitt's lymphoma at RCCH between 1984 and 2004. RESULTS: Ninety-two children were treated for Burkitt's lymphoma at RCCH between 1984 and 2004. There were 10 patients with group A or fully resected disease, 52 with group B or extensive localised disease, and 30 with dissemination to the bone marrow and/or central nervous system or group C disease. Protocol 1 (less intensive chemotherapy based on the COMP regimen) was used from 1984, with protocol 2 (more intensive chemotherapy based on the LMB regimen) introduced in 1988 for group C disease, 1991 for group B disease and 1996 for group A disease. Overall 5-year survival increased from 20% with protocol 1 to 66% with protocol 2 for group C disease, and from 76.5% with protocol 1 to 88.2% with protocol 2 for group B disease. There were more admissions for neutropenic fever in patients on protocol 2 and more episodes of mucositis, and these patients required more red cell and platelet transfusions. With a more conservative surgical approach, biopsy largely replaced attempts to partially resect the tumour at primary surgery, and there was a consequent decline in surgical complications. CONCLUSIONS: Intensive chemotherapy with protocol 2 has resulted in improved survival for group C and group B patients, but with more morbidity. Protocol 1, which is less intensive with less morbidity, remains a viable strategy for group A and group B disease in resource-poor settings. PMID- 17077924 TI - Overview of a paediatric renal transplant programme. AB - INTRODUCTION: Renal transplantation is the therapy of choice for children with end-stage renal failure. There are many challenges associated with a paediatric programme in a developing country where organs are limited. METHODS: A retrospective review was undertaken of 149 paediatric renal transplants performed between 1968 and 2006 with specific emphasis on transplants performed in the last 10 years. Survival of patients and grafts was analysed and specific problems related to drugs and infections were reviewed. RESULTS: On review of the total programme, 60% of the transplants have been performed in the last 10 years, with satisfactory overall patient and graft survival for the first 8 years post transplant. At this point, transfer to adult units with non-compliance becomes a significant problem. Rejection is less of a problem than previously but infection is now a bigger issue--specifically tuberculosis (TB), cytomegalovirus (CMV) and Epstein-Barr virus (EBV) infections with related complications. A wide variety of drugs are available for tailoring immunosuppression to minimise side-effects. CONCLUSION: It is possible to have a successful paediatric transplant programme in a developing country. However, to improve long-term outcomes certain issues need to be addressed, including reduction of nephrotoxic drugs and cardiovascular risk factors and providing successful adolescent to adult unit transition. PMID- 17077925 TI - Liver transplantation at Red Cross War Memorial Children's Hospital. AB - The liver transplant programme for infants and children at Red Cross War Memorial Children's Hospital is the only established paediatric service in sub-Saharan Africa. Referrals for liver transplant assessment come from most provinces within South Africa as well as neighbouring countries. PATIENTS AND METHODS: Since 1987, 81 children (range 6 months-14 years) have had 84 liver transplants with biliary atresia being the most frequent diagnosis. The indications for transplantation include biliary atresia (48), metabolic (7), fulminant hepatic failure (10), redo transplants (3) and other (16). Four combined liver/kidney transplants have been performed. Fifty-three were reduced-size transplants with donor/recipient weight ratios ranging from 2:1 to 11:1 and 32 children weighed less than 10 kg. RESULTS: Sixty patients (74%) survived 3 months-14 years post-transplant. Overall cumulative 1- and 5-year patient survival figures are 79% and 70% respectively. However, with the introduction of prophylactic intravenous ganciclovir and the exclusion of hepatitis B virus (HBV) IgG core Ab-positive donors, the 1-year patient survival is 90% and the projected 5-year paediatric survival is > 80%. Early (< 1 month) post-liver-transplant mortality was low. Causes include primary malfunction (1), inferior vena cava thrombosis (1), bleeding oesophageal ulcer (1), sepsis (1) and cerebral oedema (1). Late morbidity and mortality was mainly due to infections: de novo hepatitis B (5 patients, 2 deaths), Epstein-Barr virus (EBV)- related post-transplantation lymphoproliferative disease (12 patients, 7 deaths) and cytomegalovirus (CMV) disease (10 patients, 5 deaths). Tuberculosis (TB) treatment in 3 patients was complicated by chronic rejection (1) and TB-drug induced subfulminant liver failure (1). CONCLUSION: Despite limited resources, a successful paediatric programme has been established with good patient and graft survival figures and excellent quality of life. Shortage of donors because of infection with HBV and human immunodeficiency virus (HIV) leads to significant waiting-list mortality and infrequent transplantation. PMID- 17077926 TI - Deaths at Red Cross Children's Hospital, Cape Town 1999-2003--a study of death notification forms. AB - OBJECTIVES: The availability of cause-specific mortality data for children in South Africa is limited. Hospital-based data have the potential to contribute to understanding of the causation of childhood death in South Africa. The objectives of the study were to gain insights into the causes of death in a South African children's hospital. DESIGN: Prospective, descriptive study of death notification forms. SETTING: Red Cross War Memorial Children's Hospital, Cape Town. METHODS: Data from 1999 to 2003 were analysed by direct and underlying causes of death (using a modified Global Burden of Diseases (GBD) classification) and demographic variables. Death rates per 1 000 hospital admissions were calculated for certain common causes of death. Seasonal correlates of mortality were examined. RESULTS: There were 1 978 deaths. The number of deaths per year increased by 11.4% over the period. The death rate rose from 15.9 to 18.4 per 1 000 admissions from 1999 to 2002, declining to 17.4/1 000 in 2003. The death rate was higher for females than for males (18.4/1 000 versus 17.6/1 000, p = 0.007). Sixty per cent of deaths occurred in children less than 1 year old. GBD group I diseases (infectious, nutritional, perinatal) accounted for the greatest proportion of deaths (58.6%), followed by noncommunicable diseases (29.1%), and injuries (7.9%). HIV/AIDS accounted for 60% of infectious deaths (31.6% of all deaths). Diarrhoea-related mortality was 3 times higher in summer than in winter. Congenital conditions dominated GBD group II (57.5%). CONCLUSION: The analysis shows the value of routinely recording data on childhood hospital deaths. The results mirror those of the South African Medical Research Council's Burden of Disease studies but also reflect the hospital's tertiary functions. Female children were at higher risk of death. Childhood HIV-related deaths are a major challenge to the health system. PMID- 17077927 TI - Surgical treatment for "brain compartment syndrome" in children with severe head injury. AB - OBJECTIVES: Traumatic brain injury accounts for a high percentage of deaths in children. Raised intracranial pressure (ICP) due to brain swelling within the closed compartment of the skull leads to death or severe neurological disability if not effectively treated. We report our experience with 12 children who presented with cerebral herniation due to traumatic brain swelling in whom decompressive craniectomy was used as an emergency. DESIGN: Prospective, observational. SETTING: Red Cross Children's Hospital. SUBJECTS: Children with severe traumatic brain injury and cerebral swelling. OUTCOME MEASURES: Computed tomography (CT) scanning, ICP control, clinical outcome. RESULTS: Despite the very poor clinical condition of these children preoperatively, aggressive management of the raised pressure resulted in unexpectedly good outcomes. CONCLUSION: Aggressive surgical measures to decrease ICP in the emergency situation can be of considerable benefit; the key concepts are selection of appropriate patients and early intervention. PMID- 17077928 TI - Selective cerebral hypothermia for post-hypoxic neuroprotection in neonates using a solid ice cap. AB - OBJECTIVE: The main objective of this study was to study the safety and efficacy of a simple, cost-effective method of selective head cooling with mild systemic hypothermia in newborn infants with hypoxic ischaemic encephalopathy. DESIGN: Ethical approval was obtained for a randomized controlled study in which 20 asphyxiated neonates with clinical signs of hypoxic ischaemic encephalopathy would be randomised into cooled and non-cooled groups. However, after cooling the first 4 babies, it was clear that repeated revisions to the cooling technique had to be made which was inappropriate in the context of a randomised controlled trial. The study was therefore stopped and the data for the 4 cooled infants are presented here in the form of a technical report. Hypothermia was achieved by applying an insulated ice cap to the heads of the infants and replacing it at 2-3 hourly intervals, aiming to achieve a target rectal temperature of 35-35.5 degrees C and a target scalp temperature of 10-28 degrees C. SETTING: This study was carried out between July 2000 and September 2001 in the neonatal units of Groote Schuur Hospital and Mowbray Maternity Hospital, Cape Town. SUBJECTS: Term infants with signs of encephalopathy were recruited within the first 8 hours of life if they had required resuscitation at birth and had significant acidosis within the first hour of life. RESULTS: Target rectal temperature was achieved in all infants, but large variations in incubator and scalp temperatures occurred in 3 of the 4 infants. Reducing the target core temperature in a stepwise manner did not prevent excessive temperature variation and resulted in a longer time to reach target temperature. There was least variation in scalp temperature when the ice pack was covered in two layers of mutton cloth before application, but the resulting scalp temperatures were above the target temperature. The maximum scalp temperature variation was reduced from 22 degrees C to 12 degrees C using this method. Nasopharyngeal temperatures varied excessively within less than a minute, suggesting that air cooling via mouth breathing was occurring. The surface site that correlated best with deep rectal temperature was the back, with the infant supine. During cooling, the respiratory rate and heart rate dropped while the mean arterial blood pressure was elevated. There were no irreversible adverse events due to cooling, but infants did become agitated and exhibited shivering which required sedation and analgesia. CONCLUSIONS: Nasopharyngeal temperature monitoring was not reliable as an acute clinical indicator of brain temperature in these spontaneously breathing infants, and the back temperature in supine infants correlated better with deep rectal temperature than did exposed skin temperature. This method of cooling achieved systemic cooling but there were large variations in regional temperatures in 3 of the 4 infants. The variations in temperature were probably due to the excessive cooling effect of the ice cap, coupled with the use of external heating to maintain systemic temperature at 35 35.5 degrees C. Variation in temperature was reduced when additional insulation was provided. However, the additional insulation resulted in the loss of the selective cerebral cooling effect. This cooling technique was therefore not an appropriate method of selective head cooling, but did successfully induce systemic hypothermia. This method of insulating an ice cap could therefore be used to induce whole-body cooling but the use of lower core temperatures of 33-34 degrees C is recommended as this will probably result in fewer regional temperature fluctuations. Ideally a more uniform method of cooling should be used. PMID- 17077929 TI - Secular trends in risk behaviour of Cape Town grade 8 students. AB - OBJECTIVE: To compare prevalence rates of selected risk behaviours and age of first intercourse of grade 8 students in Cape Town between 1997 and 2004. DESIGN: Cross-sectional surveys in 1997 and 2004. Survival analysis was used to estimate the cumulative incidence of first intercourse. The log-rank statistic was used to compare the survival distributions. When comparing data from the two studies we used a logistic regression model with the factors year, race and age group to test the difference in reported risk behaviours between 1997 and 2004 within each gender. SETTING: Public high schools in Cape Town. SUBJECTS: Multistage cluster samples of 1 437 and 6 266 grade 8 students in 1997 and 2004 respectively. OUTCOME MEASURES: Ever having had sexual intercourse; for those that had, whether any method was used to prevent pregnancy or disease at last intercourse, and (if so) what was used; use of tobacco, alcohol and marijuana; violence-related behaviours; and suicidal behaviour. RESULTS: There was a significant delay in first intercourse in 2004 compared with 1997. For males, levels of condom use were lower in 2004 than in 1997, while for females levels of injectable contraceptive use were lower. There were significant increases in past month use of cigarettes for males and marijuana for both genders. Rates of perpetration of violence behaviour remained stable or decreased from 1997 to 2004, while the rate of suicidal behaviour for males increased. CONCLUSIONS: School-based interventions that address sexual risk behaviours should be expanded to include other risk behaviours. PMID- 17077930 TI - Antiretroviral treatment for children. AB - OBJECTIVE: To describe the response of children during their first year on highly active antiretroviral therapy (HAART). DESIGN: Retrospective, descriptive. SETTING: Tertiary, referral hospital. SUBJECTS: All HIV-infected children commenced on HAART from 1 August 2002 until 31 December 2004. OUTCOME MEASURES: Children were retrospectively restaged using the WHO 4-stage clinical classification and CDC immunological staging system. After commencing HAART, patients were assessed at monthly intervals for the first 6 months and thereafter mostly 3-monthly. Baseline and 6- monthly CD4 counts and viral loads were performed. RESULTS: Of 409 children commenced on HAART, 50.6% were < 2 years old, 62.7% had severe clinical disease and 76.6% had severe immune suppression. After 1 year, 65.8% were alive and continued HAART at the hospital, 11.2% had been transferred to another antiretroviral site, 15.4% had died, 4.6% were lost to follow-up and treatment had been discontinued in 2.9%. Kaplan-Meier survival estimate for 407 children at 1 year was 84% (95% confidence interval (CI) 80 - 87%). On multivariate analysis, survival was adversely affected in children with WHO stage 4 v. stage 2 and 3 disease (adjusted hazard ratio (HR): 5.26 (95% CI 2.25 - 12.32), p = 0.000), age < 12 months (adjusted HR: 2.46 (95% CI 1.48 - 4.09), p = 0.001) and CD4 absolute count (per 100 cell increase) (adjusted HR: 0.93 (95% CI 0.88 - 0.98), p = 0.013). In a separate multivariate model including only children with an initial viral load (N = 367), viral load > or = 1 million copies/ml (adjusted HR: 1.84 (95% CI 1.03 - 3.29)) and taking a protease inhibitor (PI)-based regimen (adjusted HR: 2.25 (95% CI 1.10 - 4.61)) were additionally independently associated with poorer survival; however, young age was not a significant predictor of mortality, after adjusting for viral load (p = 0.119). After 1 year of HAART 184/264 (69.7%) of children had a viral load < 400 copies/ml. Comparative analysis showed significant improvements in growth, immunological status and virological control. CONCLUSION: HAART can improve the health of many HIV-infected children with advanced disease, including those aged less than 2 years in resource-limited settings. PMID- 17077931 TI - Determining the prevalence of malnutrition in hospitalized paediatric patients. AB - AIM: To determine the prevalence of malnutrition in hospitalised paediatric patients at Red Cross War Memorial Children's Hospital. METHOD: A 1-day cross sectional survey was completed in all medical and surgical wards and some specialist outpatient clinics. RESULTS: A total of 227 children participated in the study. Thirty-five per cent of patients were moderately malnourished (< or = 2 z-score), of whom 70% had no road to health card with them. Thirty-four per cent of children under 60 months of age received supplements in addition to a normal ward diet, 7.8% were enterally fed and less than 1% were parenterally fed. Almost 14% of children were found to be overweight/obese, which is higher than the national average of 6%. The prevalence of HIV infection on the day of the audit was 18% across all age groups compared with the Western Cape antenatal prevalence of 15.7% (2005). CONCLUSION: The overall prevalence of undernutrition was 34%, which is comparable with similar studies. However, the proportion of overweight children (14%) was greater than the national average. In view of the level of malnutrition seen, a nutrition risk-screening tool, identifying risk factors for malnutrition such as food access and vulnerability, should be developed. The tool should be used to assess nutrition status and risk during the course of hospitalisation, in addition to planning appropriate nutrition care plan interventions for discharge. PMID- 17077932 TI - [Disseminated skin ulcerations associated with chronic renal failure]. PMID- 17077933 TI - [Teaching and simulation. Methods, demands, evaluation and visions]. AB - Since 1st October 2003 the new German "Approbationsordnung fur Arzte" (Medical Licensing Regulations) requires an increasing amount of small group teaching sessions and encourages a multidisciplinary and more practical approach to the related topics. In 2004 the German Society of Anaesthesiology and Intensive Care Medicine has provided almost all anaesthesia faculties of German Universities with equipment for full-scale simulation. This article describes methods for a simulation-based medical education training program. Basic requirements for a successful training program using full scale simulators are the provision of an adequate logistical and material infrastructure, teacher attendance of train-the trainer courses, implementation in the medical curriculum and an instructor student ratio of 1:3, equivalent to that for bedside teaching. If these requirements were fulfilled, medical students scored the simulation scenarios "induction of anaesthesia", "acute pulmonary embolism", "acute management of a multiple trauma patient" and "postoperative hypotension" as 1.5, 1.6, 1.5 and 1.5, respectively, on a scale of 1-6. These scores were better than those given for other segments of the curriculum. PMID- 17077934 TI - [Anaesthesia for cardiac catheterization in children]. AB - Cardiac catheterization in children with congenital heart disease or in adults with completely or partially corrected cardiac defects, is a growing field of activity for anaesthesiologists. This requires not only the willingness for interdisciplinary co-operation, but also detailed knowledge about the pathophysiology of congenital heart diseases. In interventional paediatric cardiology significant innovations have occurred during recent years including stenting of a patent ductus arteriosus and of peripheral pulmonary artery stenosis. Furthermore, radiofrequency catheter ablation for recurrent tachyarrhythmia, or resynchronisation therapy with biventricular pacing in the setting of congestive heart failure, or implantation of cardioverter defibrillators are increasingly being employed which require anaesthesia support. PMID- 17077935 TI - ["We must relate to ourselves". Substance, causality, and narrative text]. AB - Since Aristotle the concept of "substance" has described those features of an object which remain constant over time despite varying additional stochastic factors (coincidences). "Causality" places several or more chronological events into a direct relationship. The change of stochastic factors on a substance always has a causal justification. Based on these metaphysical considerations the present contemplations will apply the concept of substance to people. The causal explanation for changes during the lifetime is achieved by narrative sentences. These are, therefore, decisive for the maintenance of the identity of individuals and collectives despite all time-linked changes. The conclusion is reached that speech and thought processes based on speech patterns are essential for humans. PMID- 17077936 TI - Impact of static and dynamic A-form heterogeneity on the determination of RNA global structural dynamics using NMR residual dipolar couplings. AB - We examined how static and dynamic deviations from the idealized A-form helix propagate into errors in the principal order tensor parameters determined using residual dipolar couplings (rdcs). A 20-ns molecular dynamics (MD) simulation of the HIV-1 transactivation response element (TAR) RNA together with a survey of spin relaxation studies of RNA dynamics reveals that pico-to-nanosecond local motions in non-terminal Watson-Crick base-pairs will uniformly attenuate base and sugar one bond rdcs by approximately 7%. Gaussian distributions were generated for base and sugar torsion angles through statistical comparison of 40 RNA X-ray structures solved to <3.0 A resolution. For a typical number (>or=11) of one bond C-H base and sugar rdcs, these structural deviations together with rdc uncertainty (1.5 Hz) lead to average errors in the magnitude and orientation of the principal axis of order that are <9% and <4 degrees, respectively. The errors decrease to <5% and <4 degrees for >or=17 rdcs. A protocol that allows for estimation of error in A-form order tensors due to both angular deviations and rdc uncertainty (Aform-RDC) is validated using theoretical simulations and used to analyze rdcs measured previously in TAR in the free state and bound to four distinct ligands. Results confirm earlier findings that the two TAR helices undergo large changes in both their mean relative orientation and dynamics upon binding to different targets. PMID- 17077937 TI - Expression of MHC I and NK ligands on human CD133+ glioma cells: possible targets of immunotherapy. AB - Mounting evidence suggests that gliomas are comprised of differentiated tumor cells and brain tumor stem cells (BTSCs). BTSCs account for a fraction of total tumor cells, yet are apparently the sole cells capable of tumor initiation and tumor renewal. BTSCs have been identified as the CD133-positive fraction of human glioma, whereas their CD133-negative daughter cells have limited proliferative ability and are not tumorogenic. It is well established that the bulk tumor mass escapes immune surveillance by multiple mechanisms, yet little is known about the immunogenicity of the CD133-positive fraction of the tumor mass. We investigated the immunogenicity of CD133-positive cells in two human astrocytoma and two glioblastoma multiforme samples. Flow cytometry analyses revealed that the majority of CD133-positive cells do not express detectable MHC I or natural killer (NK) cell activating ligands, which may render them resistant to adaptive and innate immune surveillance. Incubating CD133-positive cells in interferon gamma (INF-gamma) significantly increased the percentage of CD133-positive cells that expressed MHC I and NK cell ligands. Furthermore, pretreatment of CD133 positive cells with INF-gamma rendered them sensitive to NK cell-mediated lysis in vitro. There were no consistent differences in immunogenicity between the CD133-positive and CD133-negative cells in these experiments. We conclude that CD133-posistive and CD133-negative glioma cells may be similarly resistant to immune surveillance, but that INF-gamma may partially restore their immunogenicity and potentiate their lysis by NK cells. PMID- 17077938 TI - Primary diffuse leptomeningeal oligodendrogliomatosis causing sudden death. AB - Primary diffuse leptomeningeal oligodendrogliomatosis is a rare malignancy of central nervous system without evidence of a primary intraparenchymal focus. We present a 25-year-old woman with the postmortem diagnosis of primary diffuse leptomeningeal oligodendrogliomatosis. She was paraplegic and had sensory loss at the level of thoracal 8-9 for nearly 15 months. There was no symptom due to increased intracranial pressure. Unexpectedly she died just before biopsy. Autopsy revealed a nodular lesion at the level of thoracal 10 vertebra and diffuse oligodendroglioma throughout the leptomeninges of the brain and spine without intraaxial focus. To our knowledge, this is the first case of primary diffuse leptomeningeal oligodendrogliomatosis causing sudden death in the literature. PMID- 17077939 TI - Increased expression of aquaporin 1 in human hemangioblastomas and its correlation with cyst formation. AB - Aquaporins (AQPs) is a water channel family which facilitates the passage of water across cell membranes. Recently, expression of aquapporin 1 (AQP1) was found to be involved in not only water transport but also tumorigenesis. In present study, we analyzed the expression of AQP1 in 26 consecutive cases of human hemangioblastomas. Significant upregulation of AQP1 expression was found in hemangioblastomas compared with control brain (P=0.002). In hemangioblastomas, expression of AQP1 was predominantly localized on membranes of stromal cells. The expression level of AQP1 in cystic group of hemangioblastomas is much higher than that of solid group (P=0.021). Most hemangioblastomas showed a negative expression of AQP1 on endothelial cells. These results imply that increased expression of AQP1 in stromal cells may play a role in cyst formation and tumorigenesis of heman-gioblastomas. PMID- 17077940 TI - De novo 617G-A nucleotide mutation in the ACVR1 gene in a Taiwanese patient with fibrodysplasia ossificans progressiva. AB - Fibrodysplasia ossificans progressiva (FOP) is a rare congenital disease with autosomal dominant transmission characterized by the presence of malformations of the big toes and of postnatal progressive heterotopic endochondral osteogenesis. We report the case of 3-year-old girl with dysplasia of the first metatarsal bones and progressive heterotopic ossificans of the right thigh due to previous diphtheria-tetanus-pertussis immunizations and several inappropriate surgical interventions. Direct sequence analysis identified a 617G-A nucleotide mutation in the patient but not in her parents or brother. Pedigree analysis suggests that a de novo mutation in the ACVR1 gene is responsible for the disease in this family. This is the first report of the results of a mutation analysis in a sporadic case of FOP in a Taiwanese patient. PMID- 17077941 TI - Photosynthetic and biochemical activities in flag leaves of a newly developed superhigh-yield hybrid rice (Oryza sativa) and its parents during the reproductive stage. AB - Responses of net photosynthetic rates to intercellular CO(2) concentration (P (n)/C (i) curves) and photochemical characteristics were investigated in flag leaves of newly developed superhigh-yield hybrid rice (Oryza sativa L.) LiangYouPeiJiu (LYPJ) and its maternal PeiAi64S (PA64S) and paternal WuMang9311 (WM9311) lines grown in the field during the reproductive stage. The results showed that photosynthetic functions, such as the electron transport activities of photosystems and photophosphorylation, assessed in vivo from P (n)/C (i) curves under field conditions declined more or earlier than those obtained in vitro. The degradation of polypeptides of thylakoid membranes was slower than those for P (Ca=360) (light-saturated net photosynthetic rate measured at 360 mumol mol(-1)) and CE (carboxylation efficiency, obtained from the initial slope of the P (n)/C (i) curve). The initial inhibition of the PSII electron transport and oxygen-evolving activity induced by senescence occurred before the degradation of the oxygen-evolving complex. In comparison, LYPJ had intermediate photosynthetic functions in the early stage of leaf development, but greater photochemical activities in the mid and late stages. WM9311 showed a similar pattern of changes but lower values, and PA64S had higher values in the early stage but showed a faster rate of senescence than LYPJ. These findings implied that the hybrid LYPJ demonstrated intermediate photosynthetic activities between its parents in the early stage of leaf development, whereas it had higher photosynthetic activities than its parents in the mid and late stages, which may be responsible for its high yield. PMID- 17077942 TI - Latitude, socioeconomic prosperity, mobile phones and hip fracture risk. AB - INTRODUCTION: Epidemiological observations suggest that sunlight exposure is an important determinant of hip fracture risk. The aim of this ecological study was to examine the relationship between latitude and hip fracture probability. METHODS: Hip fracture incidence and mortality were obtained from literature searches and 10-year hip fracture probability computed from fracture and death hazards. RESULTS: There was a significant association between latitude and 10 year hip fracture probability. For each 10 degrees change in latitude from the equator (e.g., from Paris to Stockholm), fracture probability increased by 0.3% in men, by 0.8% in women and by 0.6% in men and women combined. There was also a significant association between economic prosperity and hip fracture risk as judged by gross domestic product (GDP)/capita or the use of mobile phones/capita. A US $10,000 higher GDP/capita was associated with a 1.3% increase in hip fracture probability. The association between latitude and hip fracture probability persisted after adjusting for indices of economic prosperity. CONCLUSIONS: These findings provide support for an important role of sunlight exposure in the global variation of hip fracture risk. In addition, there is a need to identify the factors related to socioeconomic prosperity that may provide mechanisms for the variation in hip fracture probability worldwide. PMID- 17077943 TI - Height and bone mineral density in androgen insensitivity syndrome with mutations in the androgen receptor gene. AB - INTRODUCTION AND HYPOTHESIS: Androgen insensitivity syndrome (AIS) constitutes a natural model to study effects of androgens and estrogens on growth and bone density. We evaluated height and bone density in patients with AIS with mutations in the androgen receptor (AR) gene. METHODS: A retrospective analysis was conducted of eight subjects with complete AIS (CAIS) and four with partial AIS (PAIS) submitted to gonadectomy followed by estrogen replacement, and three with PAIS who did not undergo gonadectomy. Standing height and bone mineral apparent density (BMAD) by DXA were measured and compared with male (z (m)) and female (z (f)) reference populations. The z-scores were compared with a value of zero using the one-sample t-test. RESULTS: Final heights of patients with CAIS and PAIS were intermediate between those predicted for females and males. BMAD of the lumbar spine in CAIS and PAIS after gonadectomy and estrogen replacement (z (f) = - 1.56 +/- 1.04, P = 0.006, and z (m) = - 0.75 +/- 0.89, P = 0.04) indicated vertebral bone deficit, whereas BMAD at the femoral neck was normal. No patient reported fractures. CONCLUSION: Subjects with AIS had mean final height intermediate between mean normal male and female, and decreased bone mineral density in the lumbar spine. These data suggest an important role for androgens in normal male growth and bone density not replaced by estrogens. PMID- 17077944 TI - An anthropometric study of relationships between the clival angle and craniofacial measurements in adult human skulls. AB - STATING BACKGROUND: The clival angle, between the ethmoidal plane and the clival plane, measures the cranial base angulation. Relations between cranial base angulation and facial patterns have been discussed and present contradictory evidence. The aim of this study is to determine whether correlations could be found between the clival angle and craniofacial measurements. METHODS: This study was carried out on a sample of 235 human skulls. Twenty landmarks were marked. The clival angle and 16 other variables were measured and compared using analysis of variance. RESULTS: Statistical analysis showed a relationship between the clival angle and six measurements (P < 0.05). There was a positive correlation between the clival angle and the cranial lengths, the nasomaxillary sagittal measurements and palatal length but the width of the foramen magnum had a negative correlation. No significant relationship was found between the clival angle and the cranial height and widths, the palatal width, the length of the foramen magnum and the mandibular measurements. CONCLUSIONS: The trends found in this study with regard to the craniofacial lengths show that the nasomaxillary complex in humans has a stable position lying beneath the anterior cranial base. Transversal, vertical and mandibular variables have a more independent relationship. PMID- 17077945 TI - Common hepatic artery arising from the left gastric artery: a rare anatomic variation identified on a cadaveric liver donor. AB - Anatomical variations of the arterial supply of the liver are not uncommon. The typical normal "textbook" anatomy of the hepatic artery is found only in approximately half of the cases. Some of the variations such as the presence of a right or left hepatic branch are more common, but other ones are extremely rare. We describe here a rare case in which the common hepatic artery arose from the left gastric artery, found during a cadaveric liver donor harvesting and confirmed with imaging studies. Cases like this one highlight the importance of knowing the hepatic arterial anatomy and the possibility of its numerous variations by the transplant and hepatobiliary surgeon. PMID- 17077946 TI - Hypopituitarism. AB - Hypopituitarism is the partial or complete insufficiency of anterior pituitary hormone secretion and may result from pituitary or hypothalamic disease. The reported incidence (12-42 new cases per million per year) and prevalence (300-455 per million) is probably underestimated if its occurrence after brain injuries (30-70% of cases) is considered. Clinical manifestations depend on the extent of hormone deficiency and may be non specific, such as fatigue, hypotension, cold intolerance, or more indicative such as growth retardation or impotence and infertility in GH and gonadotropin deficiency, respectively.A number of inflammatory, granulomatous or neoplastic diseases as well as traumatic or radiation injuries involving the hypothalamic-pituitary region can lead to hypopituitarism. Several genetic defects are possible causes of syndromic and non syndromic isolated/multiple pituitary hormone deficiencies. Unexplained gonadal dysfunctions, developmental craniofacial abnormalities, newly discovered empty sella and previous pregnancy-associated hemorrhage or blood pressure changes may be associated with defective anterior pituitary function.The diagnosis of hypopituitarism relies on the measurement of basal and stimulated secretion of anterior pituitary hormones and of the hormones secreted by pituitary target glands. MR imaging of the hypothalamo-pituitary region may provide essential information. Genetic testing, when indicated, may be diagnostic.Secondary hypothyroidism is a rare disease. The biochemical diagnosis is suggested by low serum FT4 levels and inappropriately normal or low basal TSH levels that do not rise normally after TRH. L-thyroxine is the treatment of choice. Before starting replacement therapy, concomitant corticotropin deficiency should be excluded in order to avoid acute adrenal insufficiency. Prolactin deficiency is also very rare and generally occurs after global failure of pituitary function. Prolactin deficiency prevents lactation. Hypogonadotropic hypogonadism in males is characterized by low testosterone with low or normal LH and FSH serum concentrations and impaired spermatogenesis. Hyperprolactinemia as well as low sex hormone binding globulin concentrations enter the differential diagnosis. Irregular menses and amenorrhea with low serum estradiol concentration (<100 pmol/l) and normal or low gonadotropin concentrations are the typical features of hypogonadotropic hypogonadism in females. In post menopausal women, failure to detect high serum gonadotropin values is highly suggestive of the diagnosis. In males, replacement therapy with oral or injectable testosterone results in wide fluctuations of serum hormone levels. More recently developed transdermal testosterone preparations allow stable physiological serum testosterone levels. Pulsatile GnRH administration can be used to stimulate spermatogenesis in men and ovulation in women with GnRH deficiency and normal gonadotropin secretion. Gonadotropin administration is indicated in cases of gonadotropin deficiency or GnRH resistance but is also an option, in alternative to pulsatile GnRH, for patients with defective GnRH secretion. PMID- 17077947 TI - Growth hormone deficiency in the adult. AB - Growth hormone deficiency (GHD) in adults may be of either adult or childhood onset and may occur as isolated GHD or as multiple hormone deficiencies. Adult onset GHD (AoGHD) usually results from damage to the pituitary gland or hypothalamus. GH is frequently undetectable in normal subjects and thus GHD cannot be distinguished from the normal state using a single random GH measurement. In general, a stimulation test is required to recognize GHD. Insulin tolerance test (ITT) has been considered the gold standard by the most important scientific societies, although alternative tests, in particular GHRH plus arginine have been proposed as valuable alternative to ITT. The clinical syndrome associated with AoGHD is characterized by a wide array of symptoms and important chronic complications, such as cardiovascular complications, which may be responsible for an increased mortality. The rationale for GH replacement in adults GHD patients is justified by the beneficial effects on some clinical end points, such as quality of life (QoL) and cardiovascular risk factors, whereas the effects on mortality risk are still controversial. Over the recent years, guidelines on the use of rhGH as a substitution treatment in adult hypopituitarism have been issued by international (Growth hormone research society-GRS, Endocrine Society) and relevant national (National Institute of Clinical Excellence-UK, NICE) institutions. The aim of the paper is to review and discuss these guidelines. PMID- 17077948 TI - Acromegaly. AB - Acromegaly is a slowly progressive disease characterized by 30% increase of mortality rate for cardiovascular disease, respiratory complications and malignancies. The estimated prevalence of the disease is 40 cases/1000000 population with 3-4 new cases/1000000 population per year. The biochemical diagnosis is based upon the demonstration of high circulating levels of GH and IGF-I. A random GH level lower than 0.4 microg/l and an IGF-I value in the age- and sex-matched normal range makes the diagnosis of acromegaly unlikely. In doubtful cases, the lack of GH suppressibility below 1 microg/l (0.3 microg/l according to recent reports) after an oral glucose load will confirm the diagnosis. A pituitary adenoma is demonstrated in most cases by CT scan or MRI. A negative X-ray finding or the presence of empty sella do not exclude the diagnosis. Cardiovascular complications (acromegalic cardiomyopathy and arterial hypertension) should be looked for and, if present, followed-up by echocardiography and 24h-electrocardiogram. Sleep apnoea, when clinically suspicious, should be confirmed by polisomnography. At the moment of diagnosis all patients should undergo colonscopy. Lipid profile should be obtained and glucose tolerance evaluated. Surgery, radiotherapy and medical treatment represent the therapeutic options for acromegaly. The outcome of transsphenoidal surgery is far better for microadenomas (80-90%) than for macroadenomas (less than 50%), which unluckily represent more than 70% of all GH-secreting pituitary tumours. Therefore, pituitary surgery is the first line treatment for microadenomas. Medical therapy is based on GH-lowering drugs (somatostatin receptor agonists and, in some cases, dopaminergic agents) and GH receptor antagonists (pegvisomant). The former are traditionally indicated after unsuccessful surgery and while awaiting the effectiveness of radiation therapy. However, GH-lowering drugs are also used as primary therapy when surgery is contraindicated or in the case of large GH-secreting macroadenomas which are not likely to be completely removed by surgery. These compounds may also be indicated in the preoperative management of some acromegalic patients in order to lower the risk of surgical and anaesthetic complications. For the moment pegvisomant is indicated for patients resistant to the GH-lowering drugs and there is no evidence for drug-induced enlargement of the pituitary tumour. In order to avoid this possibility, however, a combination of pegvisomant and GH-lowering compound can also be conceived. With pegvisomant, IGF-I plasma levels are the marker of therapeutic efficacy and normalize in 97% of patients. Radiotherapy is employed sparingly due to the number of side effects (80% of hypopituitarism). It is indicated after unsuccessful surgical and/or medical treatment and allows the control of hormonal secretion and tumour growth in approx. 40% and 100% of cases, respectively. Acromegaly is defined as controlled when, in the absence of clinical activity, IGF-I levels are in the age- and sex-matched normal range and GH is normally suppressible by the oral glucose load. PMID- 17077949 TI - Isolated corticotrophin deficiency. AB - Isolated ACTH deficiency (IAD) is a rare disorder, characterized by secondary adrenal insufficiency (AI) with low or absent cortisol production, normal secretion of pituitary hormones other than ACTH and the absence of structural pituitary defects. In adults, IAD may appear after a traumatic injury or a lymphocytic hypophysitis, the latter possibly due to autoimmune etiology. Conversely, a genetic origin may come into play in neonatal or childhood IAD. Patients with IAD usually fare relatively well during unstressed periods until intervening events spark off an acute adrenal crisis presenting with non specific symptoms, such as asthenia, anorexia, unintentional weight loss and tendency towards hypoglycemia. Blood chemistry may reveal mild hypoglycemia, hyponatremia and normal-high potassium levels, mild anemia, lymphocytosis and eosinophilia. Morning serum cortisol below 3 microg/dl are virtually diagnostic for adrenal insufficiency. whereas cortisol values comprised between 5-18 microg/dl require additional investigations: insulin tolerance test (ITT) is considered the gold standard but-when contraindicated-high or low dose-ACTH stimulation test with serum cortisol determination provides a viable alternative. Plasma ACTH concentration and prolonged ACTH infusion test are useful in differential diagnosis between primary and secondary adrenal insufficiency. For some patients with mild, near-to-asymptomatic disease, glucocorticoid replacement therapy may not be required except during stressful events; for symptomatic patients, replacement doses i.e., mean daily dose 20 mg (0.30 mg/kg) hydrocortisone or 25 mg (0.35 mg/kg) cortisone acetate, are usually sufficient. Administration of mineralocorticoids is generally not necessary as their production is maintained. PMID- 17077950 TI - Cushing's disease. AB - Cushing's disease, i.e., pituitary ACTH-secreting adenoma causing excess glucocorticoid secretion, is a rare disease with significant mortality and morbidity. Timely diagnosis and appropriate treatment can alter the course of the disease and are therefore mandatory. First step of the diagnostic work-up is the endogenous glucocorticoid excess by measurement of urinary free cortisol, cortisol circadian rhythmicity or suppression by low doses of dexamethasone. In patients with equivocal results, second line tests, such as the dexamethasone suppressed CRH test and desmopressin stimulation, usually enable the diagnosis to be confirmed. Measurement of plasma ACTH then allows the distinction between ACTH dependent (e.g., pituitary or extrapituitary neuroendocrine tumors) and ACTH independent causes (e.g., adrenal tumors). The last step in the diagnostic algorithm is often the most fraught with problems as the distinction between Cushing's disease and ectopic ACTH secretion relies on judicious interpretation of several diagnostic procedures. Positive responses to stimulation with CRH and inhibition by high doses of dexamethasone, if concurrent, enable a pituitary origin to be established whereas conflicting results call for inferior petrosal sinus sampling, the latter to be performed in experienced centres only. Visualisation of the tumor at pituitary imaging is helpful but not required for the diagnosis, as microadenomas often remain undectected by MRI and/or CT scan and, on the other hand, visualisation of a non-secreting incidentaloma may be misleading. Surgical removal of the pituitary tumor is the optimal treatment choice and should be attempted in every patient. Surgical failures as well as relapses can be treated by radiotherapy, medical therapy or, if necessary, bilateral adrenalectomy. Finally, patients cured of Cushing's disease require long-term monitoring given the risk of relapse and clinical burden of associated ailments. PMID- 17077951 TI - Anti-androgenic therapy using oral spironolactone for acne vulgaris in Asians. AB - BACKGROUND: Few studies have addressed anti-androgenic therapy using oral spironolactone for acne in Asians. Obtaining this race-specific information is important because Westerners and Asians respond differently to hormone therapy. This study aimed to examine the efficacy and safety of oral spironolactone used to treat acne in Asians. METHODS: Spironolactone (initial dose, 200 mg/day) was administered orally to 139 Japanese patients (116 females and 23 males) with acne. Serum laboratory data, including various hormones and electrolytes, were examined for 25 of the subjects. RESULTS: Most of the female patients who completed the 20-week regimen exhibited excellent improvement (evaluated by a photographic grading scale), although some discontinued treatment because of menstrual disturbances or other reasons. The treatment was less efficacious for the males than for the females, and because gynecomastia developed in three male patients, spironolactone treatment for males was stopped. Examination of the serum of 25 patients did not identify any toxicity associated with the treatment. Drug eruptions and edema in the lower extremities were each seen in three patients. CONCLUSION: Oral spironolactone is effective and safe for the treatment of acne in Asian females, and can be a good option for severe, recurring, and widespread types of the condition. PMID- 17077952 TI - The use of metatarsal bone grafts in the treatment of saddle nose deformities. AB - The results from segmental fourth metatarsal bone reconstruction of significant nasal dorsal defects are described. Solid bone graft reconstructions frequently lead to an unnatural hardness of the nasal tip. Rib cartilage reconstructions are pliable and soft, but pose a problem because they easily undergo warpage. The operation is performed using the open approach. An outer fourth metatarsal bone graft used for the bone component extends at least two-thirds of the dorsum length. It is secured in place with a compression screw and a Kirschner wire. The cartilage component comprises an abbreviated L-strut constructed of septal cartilage. It is slotted into the metatarsal bone in a tongue-in-groove manner and sutured to it through a drill hole in the bone. The dorsal profile is completed in this manner. PMID- 17077953 TI - Modified punch technique for incomplete earlobe cleft repair. PMID- 17077954 TI - Octyl-2-cyanoacrylate adhesive for skin closure and prevention of infection in plastic surgery. AB - No complete scientific study has yet investigated the incidence of surgical-site infections in plastic surgery operations. However, it has been noted that in the case of wound infection, the aesthetic and sometimes the functional results become invalidated by delay and an alteration of the healing processes, thus necessitating surgical correction. This study aimed to evaluate the effectiveness of applying tissue adhesive on surgical wounds in plastic surgery as a protection capable of reducing the onset of infection, and to verify the existence of a statistically significant difference between treated and untreated wounds, and to determine patients' satisfaction with their cicatricial results. PMID- 17077955 TI - Use of gabapentine (neurantin) for relief of intractable pain developed after face-lift surgery. AB - Complications of rhytidectomy, technically one of the most challenging operations of the aesthetic surgery, have been widely reported in the literature and nerve injuries are one of these possible complications during facial rejuvenation surgery. Beside the injuries of motor branches of the facial nerve, complaints of numbness and development of neuromas due to sensorial nerve injuries were reported in the literature. In this particular reported case, however, complaints of a 52-year-old woman, presented with diffuse intractable pain in her left cheek radiating to the neck and behind the ear after facial rejuvenation surgery, improved dramatically by use of gabapentine. Although there was no apparent neuroma formation on her neck, MRI findings are inconclusive and she had not responded to injection of local anesthetics at all. PMID- 17077956 TI - Reduction otoplasty: correction of the large or asymmetric ear. AB - Reduction otoplasty is an uncommon procedure performed for macrotia and ear asymmetry. Techniques described in the literature for this procedure are few. The authors present their ear reduction approach that not only achieves the desired reduction effectively and accurately, but also addresses and creates the natural anatomic proportions of the ear, leaving a scar well hidden within the fold of the helix. PMID- 17077957 TI - Intraabdominal pressure in abdominoplasty patients. AB - Abdominal compartment syndrome is directly related to an increase in intraabdominal pressure (IAP), which can lead in severe cases to serious clinical consequences. Routine measurement of IAP in specific cases has been advocated by some surgical specialties. However, few studies in plastic surgery have focused on the use of IAP. The authors review the literature and describe a method of IAP analysis used for 12 patients who underwent abdominoplasty. PMID- 17077958 TI - Dysphagia secondary to osteoid osteoma of the transverse process of the second cervical vertebra. AB - Dysphagia due to bony lesions of the cervical spine is rare. Almost all reported cases have been secondary to cervical osteophyte formation. We present an unusual case of a 58-year-old male who presented with dysphagia of insidious onset. Investigations revealed osteoid osteoma arising from the transverse process of the second cervical vertebra extending anteriorly in a curvilinear manner. The surgical management of this case is discussed in this report. To our knowledge, this is the first reported case of dysphagia secondary to osteoid osteoma of cervical spine. PMID- 17077959 TI - Squamous cell carcinoma of the larynx in HIV-positive patients: difficulties in diagnosis and management. AB - Patients who are infected with human immunodeficiency virus (HIV) are at increased risk of developing laryngeal squamous cell carcinoma. This malignancy on average appears in a younger age group at a more advanced stage and has a more aggressive course in HIV patients. These patients have difficult management challenges, diagnostically, in staging, and particularly in determining the optimal treatment for each individual patient because their underlying HIV infection can markedly increase morbidity associated with active treatments. They frequently have problems associated with swallowing both before and after treatment. We present two cases that highlight difficulties in the diagnosis and management of these patients as well as post-treatment complications, with particular emphasis on swallowing problems. PMID- 17077960 TI - Stabilizing selection of protein function and distribution of selection coefficient among sites. AB - In this study, I take a new approach to modeling the evolutionary constraint of protein sequence, introducing the stabilizing selection of protein function into the nearly-neutral theory. In other words, protein function under stabilizing selection generates the evolutionary conservation at the sequence level. With the help of random mutational effects of nucleotides on protein function, I have derived the distribution of selection coefficient among sites, called the S distribution whose parameters have clear biological interpretations. Moreover, I have studied the inverse relationship between the evolutionary rate and the effective population size, showing that the number of molecular phenotypes of protein function, i.e., independent components in the fitness of the organism, may play a key role for the molecular clock under the nearly-neutral theory. These results are helpful for having a better understanding of the underlying evolutionary mechanism of protein sequences, as well as human disease-related mutations. PMID- 17077961 TI - Peptidology: short amino acid modules in cell biology and immunology. AB - Short amino acid motifs, either linear sequences or discontinuous amino acid groupings, can interact with specific protein domains, so exerting a central role in cell adhesion, signal transduction, hormone activity, regulation of transcript expression, enzyme activity, and antigen-antibody interaction. Here, we analyze the literature for such critical short amino acid motifs to determine the minimal peptide length involved in biologically important interactions. We report the pentapeptide unit as a common minimal amino acid sequence critically involved in peptide-protein interaction and immune recognition. The present survey may have implications in defining the dimensional module for peptide-based therapeutical approaches such as the development of novel antibiotics, enzyme inhibitors/activators, mimetic agonists/antagonists of neuropeptides, thrombolitic agents, specific anti-viral agents, etc. In such a therapeutical context, it is of considerable interest that low molecular weight peptides can easily cross biological barriers, are less susceptible to protease attacks, and can be administered at high concentrations. In addition, small peptides are a rational target for strategies aimed at antigen-specific immunotherapeutical intervention. As an example, specific short peptide fragments might be used to elicit antibodies capable of reacting with the full-length proteins containing the peptide fragment's amino acid sequence, so abolishing the risk of cross reactivity. PMID- 17077962 TI - Non-sufficient cell cycle control as possible clue for the resistance of human malignant glioma cells to clinically relevant treatment conditions. AB - OBJECTIVES: Human gliomas have a catastrophic prognosis with a median survival in the range of one year even after therapeutic treatment. Relatively high resistance towards apoptotic stimuli is the characteristic feature of malignant gliomas. Since cell cycle control has been shown to be the key mechanism controlling both apoptosis and proliferation, this study focuses on DNA damage analysis and protein expression patterns of essential cell cycle regulators P53 and P21waf1/cip1 in glioma under clinically relevant therapeutic conditions. MATERIAL AND METHODS: U87MG cell line, characterised by wild p53-phenotype relevant for the majority of primary malignant glioblastomas, was used. Glioma cells underwent either irradiation or temozolomide treatment alone, or combined radio/chemo treatment. DNA damage was analysed by the "Comet Assay". Expression rates of target proteins were analysed using "Western-Blot" technique. RESULTS AND CONCLUSIONS: "Comet Assay" demonstrated extensive DNA damage caused by temozolomide treatment alone and in combination with irradiation, correlating well with the low survival rate observed under these treatment conditions. In contrast, irradiation alone resulted in a relatively low DNA damage, correlating well with a high survival rate and indicating a poor therapeutic efficiency of irradiation alone. Unusually low up-regulation of P53 and P21waf1/cip1 expression patterns was produced by the hereby tested stressful conditions. A deficit in cell cycle control might be the clue to the high resistance of malignant glioma cells to established therapeutic approaches. PMID- 17077963 TI - Purification of branched-chain amino acid aminotransferase from Helicobacter pylori NCTC 11637. AB - Branched-chain amino acid aminotransferase was purified by several column chromatographies from Helicobacter pylori NCTC 11637, and the N-terminal amino acid sequence was analyzed. The enzyme gene was sequenced based on a putative branched-chain amino acid aminotransferase gene, ilvE of H. pylori 26695, and the whole amino acid sequence was deduced from the nucleotide sequence. The enzyme existed in a homodimer with a calculated subunit molecular weight (MW) of 37,539 and an isoelectric point (pI) of 6.47. The enzyme showed high affinity to 2 oxoglutarate (K (m) = 0.085 mM) and L-isoleucine (K (m) = 0.34 mM), and V (max) was 27.3 micromol/min/mg. The best substrate was found to be L-isoleucine followed by L-leucine and L-valine. No activity was shown toward the D enantiomers of these amino acids. The optimal pH and temperature were pH 8.0 and 37 degrees C, respectively. PMID- 17077964 TI - Protein-carbonyl accumulation in the non-replicative senescence of the methionine sulfoxide reductase A (msrA) knockout yeast strain. AB - The major enzyme of the methionine sulfoxide reductase (Msr) system is MsrA. Senescing msrA knockout mother yeast cells accumulated significant amounts of protein-carbonyl both at 5 generation-old (young) and 21 generation-old (old) cultures, while the control mother cells showed significant levels of protein carbonyl mainly in the old culture. The Msr activities of both yeast strains declined with age and exposure of cells to H(2)O(2) caused an accumulation of protein-carbonyl especially in the msrA knockout strain. It is suggested that a compromised MsrA activity may serve as a marker for non-replicative aging. PMID- 17077965 TI - The effect of extractants on degradation of L-glutamate and L-arginine in the course of shaking and filtration at low temperature. AB - The effects of demineralized water (DEMI H(2)O) and 0.5 M ammonium acetate (0.5 M AAc) on losses of L-glutamic acid and L-arginine in the course of shaking and filtration at low temperature (6 degrees C) were tested. The concentration of L glutamic acid decreased by 6.3% in DEMI H(2)O and by 4.9% in 0.5 M AAc, whereas the L-arginine concentration decreased by 6.0% (DEMI H(2)O) and 10.7% (0.5 M AAc). We found a significantly (P < 0.05) higher degradation of L-arginine in 0.5 M AAc compared with that of DEMI H(2)O. PMID- 17077966 TI - Protein tyrosine nitration in hydrophilic and hydrophobic environments. AB - In this review we address current concepts on the biological occurrence, levels and consequences of protein tyrosine nitration in biological systems. We focused on mechanistic aspects, emphasizing on the free radical mechanisms of protein 3 nitrotyrosine formation and critically analyzed the restrictions for obtaining large tyrosine nitration yields in vivo, mainly due to the presence of strong reducing systems (e.g. glutathione) that can potently inhibit at different levels the nitration process. Evidence is provided to show that the existence of metal catalyzed processes, the assistance of nitric oxide-dependent nitration steps and the facilitation by hydrophobic environments, provide individually and/or in combination, feasible scenarios for nitration in complex biological milieux. Recent studies using hydrophobic tyrosine analogs and tyrosine-containing peptides have revealed that factors controlling nitration in hydrophobic environments such as biomembranes and lipoproteins can differ to those in aqueous compartments. In particular, exclusion of key soluble reductants from the lipid phase will more easily allow nitration and lipid-derived radicals are suggested as important mediators of the one-electron oxidation of tyrosine to tyrosyl radical in proteins associated to hydrophobic environments. Development and testing of hydrophilic and hydrophobic probes that can compete with endogenous constituents for the nitrating intermediates provide tools to unravel nitration mechanisms in vitro and in vivo; additionally, they could also serve to play cellular and tissue protective functions against the toxic effects of protein tyrosine nitration. PMID- 17077967 TI - Role of laboratory variables in differentiating SARS-coronavirus from other causes of community-acquired pneumonia within the first 72 h of hospitalization. AB - The Centers for Disease Control and Prevention (CDC) recommend that SARS coronavirus (SARS-CoV) testing be considered in epidemiologically high-risk patients hospitalized with community-acquired pneumonia (CAP) if no alternative diagnosis is identified after 72 h. The aim of this study was to identify routine laboratory variables that might indicate the need for SARS-CoV testing. Routine hematological/biochemical variables in patients with laboratory-confirmed SARS (2003) were compared with those in consecutive patients hospitalized June December 2004 with radiologically confirmed CAP. Stepwise logistic regression analyses were performed to identify discriminating variables at baseline and by day 3 of hospitalization. Nasopharyngeal aspiration and antigen detection for influenza virus and respiratory syncytial virus using an immunofluorescence assay (IFA) were routinely performed in patients with CAP. Altogether, 181 patients with CAP (who remained undiagnosed by IFA) and 303 patients with SARS were studied. The mean intervals from symptom onset to admission were 3.1 and 2.8 days, respectively (p > 0.05). The etiological agent of CAP was identified retrospectively in only 39% of cases, the majority being bacterial pathogens. At baseline, age and absolute neutrophil count (ANC) were the only independent discriminating variables (p < 0.0001). Using a value of <4.4 x 10(9)/l as the cutoff for ANC, the sensitivity and specificity of ANC for discriminating SARS were 64 and 95%, respectively (AUC 0.90). By day 3 of hospitalization, age (p < 0.0001), change in ANC (p = 0.0003), and change in bilirubin (p = 0.0065) were discriminating variables. A model combining age <65 years, a change in ANC of >-3 x 10(9)/l, and a change in bilirubin of > or =0 mmol/l had a sensitivity of 43% and a specificity of 95% for SARS (AUC 0.90). There are only a few laboratory features (including lymphopenia) that clearly discriminate SARS from other causes of CAP. Nevertheless, when evaluating epidemiologically high-risk patients with CAP and no immediate alternative diagnosis, a low ANC on presentation along with poor clinical and laboratory responses after 72 h of antibiotic treatment may raise the index of suspicion for SARS and indicate a need to perform SARS-CoV testing. PMID- 17077968 TI - [MDCT classification of osseous ankle and foot injuries]. AB - Conventional radiography plays an essential role in the primary evaluation of acute ankle and foot trauma. In the case of complex injuries, however, subsequent computed tomography (CT) is nowadays recommended. In this connection, multidetector computed tomography (MDCT) allows better temporal, spatial, and contrast resolution compared with the conventional single-slice spiral CT. Multiplanar reformation and three-dimensional reconstruction of the acquired data sets are also helpful tools for critical assessment of therapeutic intervention. This report reviews the potential of the MDCT technique for accurate fracture classification, precise illustration of displaced components, and postoperative control of arrangement of typical lesions. PMID- 17077969 TI - In situ electrochemical contact angle study of hemoglobin and hemoglobin-Fe3O4 nanocomposites. AB - The electrochemical redox-induced contact angle changes of hemoglobin droplets in the absence and presence of tetraheptylammonium-capped Fe3O4 nanoparticles have been explored by using in situ electrochemical contact angle measurements. The results indicate that the electrochemical redox process may lead to some structure changes of hemoglobin (Hb), which could further induce the hydrophobic to-hydrophilic changes of the relative droplets. Our observations demonstrate that hemoglobin could self-assemble on the surface of the functionalized Fe3O4 nanoparticles as Hb-Fe3O4 nanocomposites, which may contribute to much more significant change of the electrochemical redox-induced contact angle values than that with free nano-Fe3O4. These results suggest that in situ electrochemical contact angle measurements could be readily applied as a new and convenient method to detect some specific biological process. PMID- 17077970 TI - Trends in monitoring pharmaceuticals and personal-care products in the aquatic environment by use of passive sampling devices. PMID- 17077971 TI - Loss of Catalase-1 (Cat-1) results in decreased conidial viability enhanced by exposure to light in Neurospora crassa. AB - Light is one of the most important factors inducing morphogenesis in Neurospora crassa. The reception of light triggers the generation of reactive oxygen species (ROS) including hydrogen peroxide (H(2)O(2)). Catalase-1 (Cat-1) is one of three catalases known to detoxify H(2)O(2) into water and oxygen. We reported that the photomorphogenetic characteristics of mutants in nucleoside diphosphate kinase-1 (NDK-1), a light signal transducer, are severely affected, and NDK-1 interacted with Cat-1 in a yeast two-hybrid assay. To disclose the function of Cat-1, we created a Cat-1 loss-of-function mutant (cat-1 ( RIP )) by the repeat induced point-mutation (RIPing) method. No Cat-1 activity was detected in the mutant strain. Forty guanines were replaced with adenines in the cat-1 gene of cat-1 ( RIP ), which caused 30 amino acid substitutions. The mutant strain grew normally, but its conidia and mycelia were more sensitive to H(2)O(2) than those of the wild type. The lack of Cat-1 activity also caused a significant reduction in the conidial germination rate. Furthermore, light enhanced this reduction in cat-1 ( RIP ) more than that in the wild type. Introduction of cat-1 into the mutant reversed all of these defective phenotypes. These results indicate that Cat-1 plays an important role in supporting the survival of conidia under oxidative and light-induced stress. PMID- 17077973 TI - Unusual histological findings in a child with idiopathic nephrotic syndrome. AB - We report a 2-year-old child with corticosteroid-resistant nephrotic syndrome whose renal biopsy revealed light microscopically normal glomeruli but highly unusual finger-like projections and arches of the glomerular basement membrane. To our knowledge, the association between nephrotic syndrome and this patient's rare ultrastuctural lesion has not been previously documented in the literature. It is not certain whether this basement membrane is pathogenic or a striking but coincidental developmental anomaly. PMID- 17077972 TI - Gene transcript and metabolite profiling of elicitor-induced opium poppy cell cultures reveals the coordinate regulation of primary and secondary metabolism. AB - Elicitor-induced sanguinarine accumulation in opium poppy (Papaver somniferum) cell cultures provides a responsive model system to profile modulations in gene transcripts and metabolites related to alkaloid biosynthesis. An annotated expressed sequence tag (EST) database was assembled from 10,224 random clones isolated from an elicitor-treated opium poppy cell culture cDNA library. The most abundant ESTs encoded defense proteins, and enzymes involved in alkaloid metabolism and S-adenosylmethionine-dependent methyl transfer. ESTs corresponding to 40 enzymes involved in the conversion of sucrose to sanguinarine were identified. A corresponding DNA microarray was probed with RNA from cell cultures collected at various time-points after elicitor treatment, and compared with RNA from control cells. Several diverse transcript populations were coordinately induced, with alkaloid biosynthetic enzyme and defense protein transcripts displaying the most rapid and substantial increases. In addition to all known sanguinarine biosynthetic gene transcripts, mRNAs encoding several upstream primary metabolic enzymes were coordinately induced. Fourier transform-ion cyclotron resonance-mass spectrometry was used to characterize the metabolite profiles of control and elicitor-treated cell cultures. Principle component analysis revealed a significant and dynamic separation in the metabolome, represented by 992 independent detected analytes, in response to elicitor treatment. Identified metabolites included sanguinarine, dihydrosanguinarine, and the methoxylated derivatives dihydrochelirubine and chelirubine, and the alkaloid pathway intermediates N-methylcoclaurine, N-methylstylopine, and protopine. Some of the detected analytes showed temporal changes in abundance consistent with modulations in the profiles of alkaloid biosynthetic gene transcripts. PMID- 17077974 TI - Management of ejaculatory duct obstruction: etiology, diagnosis, and treatment. AB - Abnormalities of the distal ejaculatory ducts related to infertility have been well-documented. Although there are no specific findings associated with ejaculatory duct obstruction, several clinical findings are highly suggestive. A diagnosis of ejaculatory duct obstruction is suggested in an infertile male with oligospermia or azoospermia with low ejaculate volume, normal secondary sex characteristics, testes, and hormonal profile, and dilated seminal vesicles, midline cyst, or calcifications on TRUS. Other causes of infertility may be concomitantly present and need to be evaluated and treated. Trans urethral resection of ejaculatory ducts (TURED) has resulted in marked improvement in semen parameters, and pregnancies have been achieved. Proper patient selection and surgical experience are necessary to obtain optimal results. In case of testicular dysfunction, chances of success are minimal. Extended follow-up periods are needed after TURED to examine the long-term effects of this procedure. Better understanding of the anatomy and pathology of the ejaculatory ducts will continue to refine diagnostic and therapeutic procedures for this disorder. PMID- 17077975 TI - Use of nomograms as predictive tools in bladder cancer. AB - Bladder cancer is a common genitourinary malignancy that demonstrates a great variation in risk of tumor recurrence and progression following treatment. The dramatic differences in clinical behavior dictate vastly differing treatments, which may range from simple surveillance to combination radical surgery with systemic chemotherapy. For non-muscle invasive bladder cancer prediction of the risk of recurrence and progression is necessary to assess the need for intravesical therapy and possible early cystectomy. In contrast, prediction of advanced disease response to primary treatment such as cystectomy and the response to systemic chemotherapy plays an important role in treatment assignment for patients with muscle invasive disease. To estimate these risk traditional risk grouping schemes such as the present TNM staging system has been used to guide patient treatment. More recently, improved prognostic tools such as nomograms have been developed to provide a more accurate assessment of outcomes. Clinicians are enthusiastically working to utilize these statistical methods in bladder cancer. We summarize the current status of outcome predictive models for bladder cancer; and focus particularly on the ability of nomograms to predict disease recurrence, progression, and patient survival. PMID- 17077976 TI - [From gene to therapy. Hereditary fever syndromes gout and inflammation]. PMID- 17077977 TI - Resonance tuning in a neuro-musculo-skeletal model of the forearm. AB - In rhythmic movements, humans activate their muscles in a robust and energy efficient way. These activation patterns are oscillatory and seem to originate from neural networks in the spinal cord, called central pattern generators (CPGs). Evidence for the existence of CPGs was found for instance in lampreys, cats and rats. There are indications that CPGs exist in humans as well, but this is not proven yet. Energy efficiency is achieved by resonance tuning: the central nervous system is able to tune into the resonance frequency of the limb, which is determined by the local reflex gains. The goal of this study is to investigate if the existence of a CPG in the human spine can explain the resonance tuning behavior, observed in human rhythmic limb movement. A neuro-musculo-skeletal model of the forearm is proposed, in which a CPG is organized in parallel to the local reflexloop. The afferent and efferent connections to the CPG are based on clues about the organization of the CPG, found in literature. The model is kept as simple as possible (i.e., lumped muscle models, groups of neurons are lumped into half-centers, simple reflex model), but incorporates enough of the essential dynamics to explain behavior-such as resonance tuning-in a qualitative way. Resonance tuning is achieved above, at and below the endogenous frequency of the CPG in a highly non-linear neuro- musculo-skeletal model. Afferent feedback of muscle lengthening to the CPG is necessary to accomplish resonance tuning above the endogenous frequency of the CPG, while feedback of muscle velocity is necessary to compensate for the phase lag, caused by the time delay in the loop coupling the limb to the CPG. This afferent feedback of muscle lengthening and velocity represents the Ia and II fibers, which-according to literature-is the input to the CPG. An internal process of the CPG, which integrates the delayed muscle lengthening and feeds it to the half-center model, provides resonance tuning below the endogenous frequency. Increased co-contraction makes higher movement frequencies possible. This agrees with studies of rhythmic forearm movements, which have shown that co-contraction increases with movement frequency. Robustness against force perturbations originates mainly from the CPG and the local reflex loop. The CPG delivers an increasing part of the necessary muscle activation for increasing perturbation size. As far as we know, the proposed neuro-musculo-skeletal model is the first that explains the observed resonance tuning in human rhythmic limb movement. PMID- 17077978 TI - Evaluation of an improved technique for lumen path definition and lumen segmentation of atherosclerotic vessels in CT angiography. AB - Vessel image analysis is crucial when considering therapeutical options for (cardio-) vascular diseases. Our method, VAMPIRE (Vascular Analysis using Multiscale Paths Inferred from Ridges and Edges), involves two parts: a user defines a start- and endpoint upon which a lumen path is automatically defined, and which is used for initialization; the automatic segmentation of the vessel lumen on computed tomographic angiography (CTA) images. Both parts are based on the detection of vessel-like structures by analyzing intensity, edge, and ridge information. A multi-observer evaluation study was performed to compare VAMPIRE with a conventional method on the CTA data of 15 patients with carotid artery stenosis. In addition to the start- and endpoint, the two radiologists required on average 2.5 (SD: 1.9) additional points to define a lumen path when using the conventional method, and 0.1 (SD: 0.3) when using VAMPIRE. The segmentation results were quantitatively evaluated using Similarity Indices, which were slightly lower between VAMPIRE and the two radiologists (respectively 0.90 and 0.88) compared with the Similarity Index between the radiologists (0.92). The evaluation shows that the improved definition of a lumen path requires minimal user interaction, and that using this path as initialization leads to good automatic lumen segmentation results. PMID- 17077979 TI - Time for a common standard for cost-effectiveness in Europe? PMID- 17077980 TI - [Heart rate reduction as a therapeutic strategy: novel options]. AB - Elevated heart rate is associated with increased cardiovascular mortality. Heart rate reduction optimises myocardial oxygen consumption and decreases angina pectoris symptoms. Thus, heart rate control is an important therapeutic strategy in coronary artery disease and, for example, chronic heart failure. The pacemaker current I(f) plays a central role in determining spontaneous activity of the sinus node. Ivabradine, a selective inhibitor of the I(f) channel, reduces heart rate without any effect on cardiac contractility and without lowering blood pressure. While beta-blockers remain the first choice for heart rate reduction, in cases of adverse effects ivabradine may be used to treat stable angina pectoris. Studies evaluating possible further uses, for example in heart failure or after acute myocardial infarction, are still warranted. PMID- 17077981 TI - [Treatment of cholestatic hepatic diseases: more than the substitution of fat soluble vitamins?]. AB - The clinical-biochemical syndrome of cholestasis is characterized by an alteration in bile constituents. As a consequence, the concentrations of bilirubin, bile acids, phospholipids and cholesterol are elevated. The main clinical symptoms of cholestasis are icterus and pruritus, and in severe cases xanthelasma and xanthoma. Primary intrahepatic cholestasis, caused by impaired bile secretion in the liver, should be separated from the extrahepatic secondary cholestasis which is a consequence of a biliary obstruction. This paper evaluates the therapy of liver diseases which developed as consequence of a primary disturbance in bile secretion. PMID- 17077982 TI - Suppression of colorectal tumor growth by regulated survivin targeting. AB - A major goal in cancer gene therapy is to develop efficient gene transfer protocols that allow tissue-specific and tightly regulated expression of therapeutic genes. The ideal vector should efficiently transduce cancer cells with minimal toxicity on normal tissues and persistently express foreign genes. One of the most promising regulatory systems is the mifepristone/RU486-regulated system, which has much lower basal transcriptional activity and high inducibility. In this work, we modified this system by incorporating a cancer specific promoter, the human telomerase reverse transcriptase (hTERT) promoter. By utilizing hTERT promoter to control the regulator, RU486 could specifically induce the expression of foreign genes in cancer cells but not in normal cells. In the context of this system, a dominant negative mutant of survivin (surDN) was controllably expressed in colorectal tumor cells. The surDN expression induced by RU486 showed a dosage- and time-dependent pattern. Regulated expression of surDN caused caspase-dependent apoptosis in colorectal tumor cells but had little effect on normal cells. Analysis of cell viability showed that RU486-induced expression of surDN suppressed colorectal tumor cell growth and had synergic effect in combination with chemotherapeutic agents. The potential of this system in cancer therapy was evaluated in experimental animals. Tumor xenograft models were established in nude mice with colorectal tumor cells, and RU486 was intraperitoneally administered. The results showed that conditional expression of surDN efficiently inhibited tumor growth in vivo and prolonged the life of tumor burdened mice. Synergized with the chemotherapeutic drug cisplatin, regulated surDN expression completely suppressed tumor growth. These results indicated that this modified RU486-regulated system could be useful in cancer-targeting therapy. PMID- 17077983 TI - Brain abscess in a neonate: an unusual presentation. AB - BACKGROUND: Neonatal brain abscesses are very rare and their clinical presentation is specific for this age group. They usually occur as a complication of bacterial meningitis or septicemia. They are most often caused by gram negative organisms, and mortality and morbidity are still significant in this particular group of patients in spite of antibiotics and modern radiological tools. METHODS: We report an unusual case of a multiloculated brain abscess in a term neonate caused by Staphylococcus aureus. The abscess developed in the absence of trauma, prior surgery, cyanotic heart disease, mother's disease, or immune defect. The onset of infection in this case was not clear and image features mimicked a brain tumor in the initial evaluation. The infant was successfully treated by primary surgical excision of the lesion and a 6-week total course of intravenous antibiotics. CONCLUSION: The interest of this case lies in the rarity of the causative organism and the atypical features of clinical and neurological images in a term neonate. PMID- 17077984 TI - Epithelioid hemangioendothelioma mimicking a nerve sheath tumor clinically and on MR imaging. AB - Epithelioid hemangioendothelioma is a tumor of the soft tissues arising from the vascular endothelium. It is considered an intermediate grade malignancy. A 42 year-old female patient presented with pain and tingling down her right arm and a mass at the right medial upper extremity. MRI revealed an oblong mass along the course of the neurovascular bundle. Given the clinical and MR findings, a nerve sheath tumor was suspected. At surgery, the mass was adherent to both the brachial artery medially and the median nerve posteriorly. Pathology revealed epithelioid hemangioendothelioma. The imaging characteristics of epithelioid hemangioendothelioma on ultrasound, CT, and MRI are reviewed. Epithelioid hemangioendothelioma can mimic a nerve sheath tumor clinically and radiologically and should be considered in the differential diagnosis of tumors involving or adjacent to a neurovascular bundle. PMID- 17077985 TI - Ubiquitous expression of mRFP-1 in vivo by site-directed transgenesis. AB - Progress in our understanding of the molecular cellular basis of immune function depends on our ability to track and image individual immune cells in vivo. To this end, the development of mouse models over-expressing various fluorescent proteins would represent an important experimental tool. In this report, we describe the generation and characterization of pUbi-mRFP-1 transgenic mice, in which the monomeric form of red fluorescent protein is ubiquitously expressed in various lymphoid and non-lymphoid tissues. Our newly generated pUbi-mRFP-1 mice are unique among previously reported mice transgenic for red fluorescent proteins because a single-copy of the mRFP-1 transgene driven by human ubiquitin C promoter has been integrated by homologous recombination into the mouse hypoxanthine-guanine phosphoribosyltransferase (HPRT) locus. We show that the distinct and uniform levels of mRFP-1 expression allow easy identification of transferred hematopoietic cells by FACS analysis or confocal microscopy, even when the transferred population represents a very small proportion in the target organ. Also, even in long-term experiments, we have seen no evidence of rejection of transferred pUbi-mRFP-1 lymphocytes. Due to its far-red spectrum, mRFP-1 is an ideal partner for dual imaging with green fluorescent proteins. We observed a good visual separation between donor lymphocytes derived from either mRFP-1 or eGFP transgenic mice in recipient animals. Our study suggests that the new pUbi mRFP-1 transgenic mouse strain offers new opportunities for studying cellular interactions and migratory patterns of cells, especially for dual imaging of different cell types. In summary, our results demonstrate that a controlled strategy of transgenesis provides an effective means of ubiquitously expressing fluorescent proteins in vivo. PMID- 17077986 TI - Ultrastructural morphometric analysis of ameloblasts exposed to fluoride during tooth development. AB - Since a considerable amount of the world population is exposed to high doses of fluoride, it is of special concern to investigate its action mechanisms during dental enamel development. In this study, the toxicity of fluoride in ameloblasts during enamel development was evaluated by means of ultrastructural morphometric analysis. A total of 18 male Wistar rats were distributed into three groups. In Group I, the animals received deionized drinking water ad libitum (negative control) and in Groups II and III, they received sodium fluorided (NaF) drinking water at doses of 7 and 100 ppm ad libitum, respectively, for 6 weeks. Morphometric data were expressed as volume density of the most significant organelles present in the secretory and maturation phases of amelogenesis such as RER, granules, lysosomes, phagic vacuoles, microfilaments and mitochondria. The results showed that the volume density of mitochondria in the 100 ppm experimental group was 29% (P < 0.05) higher than the control group in secretory ameloblasts. No remarkable differences were found in maturation ameloblasts for all organelles evaluated. Taken together, these data indicate that NaF at high doses is able to induce cellular damage in secretory ameloblasts, whereas no noxious effect was observed during maturation stage of amelogenesis as depicted by ultrastructural analysis. PMID- 17077987 TI - Using a sampling strategy to address psychometric challenges in tutorial-based assessments. AB - INTRODUCTION: Tutorial-based assessment, despite providing a good match with the philosophy adopted by educational programmes that emphasize small group learning, remains one of the greatest challenges for educators working in this context. The current study was performed in an attempt to assess the psychometric characteristics of tutorial-based evaluation upon adopting a multiple sampling approach that requires minimal recording of observations. METHOD: After reviewing the literature, a simple 3-item evaluation form was created. The items were "Professional Behaviour," "Contribution to Group Process," and "Contribution to Group Content." Explicit definition of these items was provided on an evaluation form. Twenty five tutors in five different programmes were asked to use the form to evaluate their students (N=169) after every tutorial over the course of an academic unit. Each item was rated using a 10-point scale. RESULTS: Cronbach's alpha revealed an appropriate internal consistency in all five programmes. Test retest reliability of any single rating was low, but the reliability of the average rating was at least 0.75 in all cases. The construct validity of the tool was supported by the observation of increasing ratings over the course of the academic unit and by the finding that more senior students received higher ratings than more junior students. CONCLUSION: Consistent with the context specificity phenomenon, the adoption of a "minimal observations often" approach to tutorial-based assessment appears to maintain better psychometric characteristics than do attempts to assess tutorial performance using more comprehensive measurement tools. PMID- 17077988 TI - GmZFP1 encoding a single zinc finger protein is expressed with enhancement in reproductive organs and late seed development in soybean (Glycine max). AB - The plant TFIIIA zinc finger proteins play important roles in plant growth and development. In this report, we isolated a single zinc finger gene, designated as GmZFP1, from soybean flowers by in silico mRNA subtraction strategy and RT-PCR. GmZFP1 is an intronless gene and encodes a protein of 210 amino acid residues with a predicted molecular mass of 23.5 KDa. GmZFP1 contains a single zinc finger domain and a DLN-box/EAR-motif at the C-terminus. To localize GmZFP1 mRNA in various tissues, semi-quantative RT-PCR assay was performed. GmZFP1 is expressed in all detected reproductive organs including flowers, developing seeds, pods, sepals, pistils with a low level, and more accumulated in petals, stamens, but not found in all detected vegetative organs with except of stem tips. Further, we found that the expression of GmZFP1 was higher in late seed development than in early seed development. To our knowledge, GmZFP1 is the first characterized gene encoding for single zinc finger protein in soybean, and may play a role as a transcriptional factor in reproductive organs development, especially functioning in petals, stamens and late developing seeds. PMID- 17077989 TI - DNA methylation, riboswitches, and transcription factor activity: fundamental mechanisms of gene-nutrient interactions involving vitamins. AB - Nutrient-gene interactions occur with a variety of nutrients including some minerals, vitamins, polyunsaturated fatty acids and other lipids. Fundamental molecular mechanisms that underlie many of the effects of nutrients on gene expression are presented herein. Two of the mechanisms described influence gene transcription: DNA methylation and transcription factor activation. Another mechanism, riboswitching, can regulate gene expression at different levels, for example, at the mRNA translation level. The first two mechanisms are widely distributed across animal phyla. Riboswitches are documented primarily in more primitive organisms, but may prove to be of wider relevance. Riboswitches are known for several vitamins; those involving thiamine are presented here. The role of folates and retinoids in DNA methylation and transcriptional factor (nuclear retinoid receptor) activities, respectively, is presented in the context of cell proliferation and differentiation, and related physiological or pathological effects during embryogenesis and cancer. PMID- 17077990 TI - Relation between phylogenetic position, lipid metabolism and butyrate production by different Butyrivibrio-like bacteria from the rumen. AB - The Butyrivibrio group comprises Butyrivibrio fibrisolvens and related Gram positive bacteria isolated mainly from the rumen of cattle and sheep. The aim of this study was to investigate phenotypic characteristics that discriminate between different phylotypes. The phylogenetic position, derived from 16S rDNA sequence data, of 45 isolates from different species and different countries was compared with their fermentation products, mechanism of butyrate formation, lipid metabolism and sensitivity to growth inhibition by linoleic acid (LA). Three clear sub-groups were evident, both phylogenetically and metabolically. Group VA1 typified most Butyrivibrio and Pseudobutyrivibrio isolates, while Groups VA2 and SA comprised Butyrivibrio hungatei and Clostridium proteoclasticum, respectively. All produced butyrate but strains of group VA1 had a butyrate kinase activity <40 U (mg protein)(-1), while strains in groups VA2 and SA all exhibited activities >600 U (mg protein)(-1). The butyrate kinase gene was present in all VA2 and SA bacteria tested but not in strains of group VA1, all of which were positive for the butyryl-CoA CoA-transferase gene. None of the bacteria tested possessed both genes. Lipase activity, measured by tributyrin hydrolysis, was high in group VA2 and SA strains and low in Group VA1 strains. Only the SA group formed stearic acid from LA. Linoleate isomerase activity, on the other hand, did not correspond with phylogenetic position. Group VA1 bacteria all grew in the presence of 200 microg LA ml(-1), while members of Groups VA2 and SA were inhibited by lower concentrations, some as low as 5 microg ml(-1). This information provides strong links between phenotypic and phylogenetic properties of this group of clostridial cluster XIVa Gram-positive bacteria. PMID- 17077991 TI - Maternal use of oral contraceptives and risk of hypospadias - a population-based case-control study. AB - The aim of this population-based case-control study was to examine the risk of isolated hypospadias in boys born to mothers who have used oral contraceptives in early pregnancy. The study was based on data from the Hungarian Case-Control Surveillance of Congenital Abnormalities from 1980 to 1996, and included 3,038 boys with hypospadias (cases), 24,799 boys without congenital abnormalities (CA free controls), and 11,881 boys with abnormalities other than hypospadias. We used unconditional logistic regression to adjust for birth order, maternal age, maternal employment status, maternal diabetes, and pre-eclampsia. When comparing cases with CA-free controls the OR for maternal use of OC was 1.21 (95% CI: 0.67 2.17). When comparing cases with boys with other abnormalities, the OR for maternal use of OC was 0.83 (95% CI: 0.46-1.50). Our data showed that self reported maternal use of oral contraceptives during pregnancy was not associated with an increased risk of hypospadias in the offspring. PMID- 17077992 TI - Gender differences in the association between westernization and metabolic risk among Greenland Inuit. AB - BACKGROUND: The Inuit have gone through an accelerated process of modernization especially since 1950. Primarily because of the dietary transition, westernisation is expected to influence the Inuit's metabolic risk in a negative way with respect to cardiovascular risk. The aim was to analyze metabolic risk factors among Inuit in Greenland and Denmark and their relation to westernization. METHODS: 1173 adult Inuit participated in a health survey in Greenland and Denmark. The examination included a 75 g OGTT. BMI, waist-to-hip ratio, and blood pressure were measured. P-glucose, s-insulin, lipids and urine albumin/creatinine ratio were analysed. Westernization was estimated by place of residence and language. RESULTS: The prevalence of the metabolic syndrome was 20.3% among men and 19.5% among women (p = 0.73). The association between the metabolic syndrome and westernization was different for men and women. For men there was an increase in prevalence of the metabolic syndrome with westernization within Greenland, but the variation was less pronounced than the difference between the migrants and the Inuit in Greenland. Age, family history of diabetes, and non-smoking were directly associated with the metabolic syndrome, whereas high physical activity was negatively associated with the metabolic syndrome. For women there was a significant negative association between westernization and the metabolic syndrome among the three population groups in Greenland, whereas the prevalence was not significantly lower among female migrants compared with Inuit women in Greenland. Age, family history of diabetes, non-smoking, and low education were associated with the metabolic syndrome. CONCLUSIONS: The effect of westernization on metabolic risk was different for men and women. For men physical inactivity due to a decrease in subsistence hunting and fishing seems to increase the metabolic risk; for women higher education is associated with a more favorable risk profile. PMID- 17077993 TI - On Heidegger, medicine, and the modernity of modern medical technology. AB - This paper examines medicine's use of technology in a manner from a standpoint inspired by Heidegger's thinking on technology. In the first part of the paper, I shall suggest an interpretation of Heidegger's thinking on the topic, and attempt to show why he associates modern technology with danger. However, I shall also claim that there is little evidence that medicine's appropriation of modern technology is dangerous in Heidegger's sense, although there is no prima facie reason why it mightn't be. The explanation for this, I claim, is ethical. There is an initial attraction to the thought that Heidegger's thought echoes Kantian moral thinking, but I shall dismiss this. Instead, I shall suggest that the considerations that make modern technology dangerous for Heidegger are simply not in the character - the ethos - of medicine properly understood. This is because there is a distinction to be drawn between chronological and historical modernity, and that even up-to-date medicine, empowered by technology, retains in its ethos crucial aspects of a historically pre-modern understanding of technology. A large part of the latter half of the paper will be concerned with explaining the difference. PMID- 17077994 TI - Associations between the CYP17, CYPIB1, COMT and SHBG polymorphisms and serum sex hormones in post-menopausal breast cancer survivors. AB - Several polymorphisms have been identified in genes that code for enzymes involved with estrogen biosynthesis and metabolism. Little is known about the functional relevance of these polymorphisms on sex hormones in vivo. We examined the association between CYP17, CYP1B1, COMT or SHBG genotypes and serum concentrations of estrone, estradiol, free estradiol, sex hormone-binding globulin (SHBG), testosterone, free testosterone and dehyroepiandrosterone in 366 post-menopausal breast cancer survivors in New Mexico, California and Washington. Hormone levels were determined by high performance liquid chromatography and radioimmunoassay in blood drawn approximately 2 years post-diagnosis. We used generalized linear regression to calculate mean hormone levels by genotype, adjusting for age, race/ethnicity, stage, study site, tamoxifen use, number of remaining ovaries, hormone therapy use, marital status and BMI. No associations were observed between any of the genotypes and sex hormones when analyzing the main effects. In subgroup analyses, androgen levels of Hispanic women with the variant (A2) CYP17 genotype were 46-87% higher than those of women with the wild type; androgen levels were 13-20% lower in non-Hispanic whites with the variant genotype; no difference by genotype was observed for African-American women. Current tamoxifen users with the variant asn(327) SHBG genotype had 81% higher serum SHBG and 39% lower free testosterone concentrations than women with the wild-type genotype. Non-tamoxifen users with the variant SHBG allele had elevated free estradiol levels. These results provide little evidence that the CYP17, CYP1B1, and COMT polymorphisms are associated with different sex hormone levels in post-menopausal breast cancer survivors. PMID- 17077995 TI - Evaluating the organization and delivery of breast cancer services: use of performance measures to identify knowledge gaps. AB - OBJECTIVES: This paper identifies gaps in our knowledge about the quality of breast cancer care in Canada to understand where programs and resources are required to enhance health services and research capacity. METHODS: A modified Delphi approach was employed involving a 15-member multidisciplinary panel of health professionals and two rounds of rating followed by deliberation to develop evidence- and consensus-based performance measures. A literature search for Canadian health services research in breast cancer was conducted based on the indicator topics. Eligible articles were identified in indexed databases of medical literature and funded research from 1995 to 2006. RESULTS: The multidisciplinary panel selected 34 indicators spanning access to services, patient outcomes, diagnosis and staging, surgery, adjuvant therapy, pathology, and follow-up care. A total of 78 articles (66 quantitative; 12 exploratory) on these topics were reviewed. Apart from two aspects of care (communication of treatment options, supportive care), the yield of Canadian breast cancer health services research did not increase subsequent to a review conducted 10 years ago which recommended greater efforts in this area. CONCLUSIONS: Research involving quantitative and qualitative methods is needed to increase our understanding about the organization and delivery of services for breast cancer diagnosis, treatment and follow-up care. Since it is unclear how to balance competing research demands, innovative strategies are required to assemble resources for health services research on breast cancer. This could include the promotion of partnerships between researchers and policy-makers across jurisdictions, and the pooling of resources between organizations, regions or networks. PMID- 17077996 TI - Influence of 17alpha-ethynylestradiol on CYP1A, GST and biliary FACs responses in male African sharptooth catfish (Clarias gariepinus) exposed to waterborne Benzo[a]Pyrene. AB - Dose dependent effects of Benzo[a]pyrene (BaP) on cytochrome P4501A (CYP1A), glutathione-S-transferase (GST) and fluorescent aromatic compounds (FACs) metabolites biomarker responses were studied in African sharptooth catfish (Clarias gariepinus) following 24 h of waterborne exposures. Based on biomass of C. gariepinus in different tanks, BaP concentrations of 1.60, 3.44, and 18.21 microg/L that corresponded to 0.5, 1.0, and 5.0 mg/kg body weight were used. Significant induction of EROD activities in gill filaments was observed at all doses and the accumulation of FACs metabolites in bile was significantly different between groups. Accumulation of FACs metabolites in bile strongly correlated (r (2) = 0.99) with BaP doses. Hepatic EROD activities were undetectable and no effect on GST activities was observed. The highest dose of BaP from the dose dependent study was further studied to assess the interactive and temporal responses of C. gariepinus on CYP1A, GST, and FACs metabolites biomarkers following exposure to either BaP alone, 17alpha-ethynylestradiol (EE(2)) alone or a combination of both compounds at concentrations of 54.17 microg/L for BaP, 51.38 microg/L for EE(2) and 54.44 microg/L for each of both compounds. Based on biomass in each tank, these concentrations corresponded to 5 mg/kg body weight. While a group of six fish was sacrificed on day 0 from the control tank only, other groups of six fish were sacrificed after 1, 3, and 6 days of exposure from the control and exposed groups. Maximum induction of gill filament and hepatic EROD activities was observed after 1 day of exposure. Both EROD activities in gill filaments and liver were significantly induced by exposure to BaP alone or co-administration with EE(2). Gill filament EROD induction was significantly inhibited (50%) by co-administration of BaP and EE(2) compared to administration of BaP alone. Levels of FACs in bile for BaP and BaP + EE(2) exposed groups were significantly different from the control at all doses and time points. A significant induction of GST activities was observed in fish exposed to BaP and BaP + EE(2) after 3 days. Exposure to EE(2) alone caused significant induction of this enzyme after day 6. This study reports for the first time the significant antagonistic influence of EE(2) on BaP in gills of fish following waterborne exposures. The results also indicate that chemical mixtures may affect biomarker responses differently from compounds administered alone and that the sensitivity of CYP1A to interactive chemicals is different in gills and liver. PMID- 17077997 TI - A toxicity and hazard assessment of fourteen pharmaceuticals to Xenopus laevis larvae. AB - The toxicity of fourteen widely used human pharmaceuticals was determined using the Frog Embryo Teratogenesis Assay-Xenopus (FETAX). Stage 9 Xenopus blastulae were exposed for 96 h to single concentrations of commonly prescribed selective serotonin reuptake inhibitors (SSRIs), statin blood lipid regulators, non steroidal anti-inflammatories, antibiotics, a stimulant, and an anti-epileptic. Toxicity, teratogenicity, minimum concentration to inhibit growth, and types and severity of associated malformations were determined. EC(10)s ranged from 3.0 mg/l to >100 mg/l and LC(10)s ranged from 3.6 mg/l to >100 mg/l. Toxicity varied between and within compound class of pharmaceutical. The fluoroquinolones, stimulants, anti-epileptics, and antibiotics tested were determined to be nontoxic and non-teratogenic at singular, water-soluble concentrations. The hazard quotients (HQ) for the pharmaceuticals ranged from 6.10 x 10(-7 )to 1.6 x 10(-4), all of which are orders of magnitude below EPA's levels for concern for harm to aquatic animals. Thus, based on the data from the present study, concentrations of individual pharmaceuticals currently detected in surface water are far below concentrations of effective and lethal concentrations. PMID- 17077998 TI - A phase I and pharmacokinetic study of silybin-phytosome in prostate cancer patients. AB - Silibinin is a polyphenolic flavonoid isolated from milk thistle with anti neoplastic activity in several in vitro and in vivo models of cancer, including prostate cancer. Silybin-phytosome is a commercially available formulation containing silibinin. This trial was designed to assess the toxicity of high-dose silybin-phytosome and recommend a phase II dose. Silybin-phytosome was administered orally to prostate cancer patients, giving 2.5-20 g daily, in three divided doses. Each course was 4 weeks in duration. Thirteen patients received a total of 91 courses of silybin-phytosome. Baseline patient characteristics included: median age of 70 years, median baseline prostate specific antigen (PSA) of 4.3 ng/ml, and a median ECOG performance status of 0. The most prominent adverse event was hyperbilirubinemia, with grade 1-2 bilirubin elevations in 9 of the 13 patients. The only grade 3 toxicity observed was elevation of alanine aminotransferase (ALT) in one patient; no grade 4 toxicity was noted. No objective PSA responses were observed. We conclude that 13 g of oral silybin phytosome daily, in 3 divided doses, appears to be well tolerated in patients with advanced prostate cancer and is the recommended phase II dose. Asymptomatic liver toxicity is the most commonly seen adverse event. PMID- 17077999 TI - A feeding stimulant for Manduca sexta from Solanum surattenses. AB - The acceptance of Solanum surattenses as a host plant for the larvae of Manduca sexta was explained by the presence of feeding stimulants in foliage. Bioassay guided fractionation of plant extracts resulted in the isolation of a highly active compound (1), which was identified as a furostan derivative {26-O-beta-D glucopyranosyl-(25R)-furosta-5-ene-3-beta-yl-O-alpha-L-rhamnopyranosyl-(1''-2')-O alpha-L-rhamnopyranosyl-(1'''-3'')-O-beta-D-glucopyranoside}. This compound has the same steroidal core substructure as that in a stimulant (indioside D) previously identified from potato foliage. However, the sugar substituents attached to the core are different. PMID- 17078000 TI - Temporal and spatial variation of terpenoids in eastern hemlock (Tsuga canadensis) in relation to feeding by Adelges tsugae. AB - The terpenoid content of eastern hemlock (Tsuga canadensis) foliage was measured over an annual cycle of development from bud opening, shoot elongation, shoot maturation, to bud-break at the start of the next growing season. The objective was to determine if variation in terpenoid composition is linked with spatial and temporal feeding preferences of the hemlock woolly adelgid (HWA; Adelges tsugae). The HWA has two periods of feeding over the course of 1 yr spanning two complete generations. There are two periods of feeding separated by a nonfeeding period where the adelgid estivates. HWA prefers to feed on mature, rather than young, expanding tissue. Feeding occurs in the leaf cushion at the base of the needle. The needle is the only tissue in hemlock with resin canals that store terpenoids. The needle and leaf cushion of both the current and previous years' growth were analyzed separately over a 1-yr period to examine the variation of terpenoid composition in space and time. Terpenoids were quantified by using headspace solid-phase microextraction/gas chromatography/mass spectrometry (SPME/GC/MS). New growth needles and leaf cushions do not resemble the previous year's growth either visually or in chemical composition until October/November, when the adelgid breaks estivation and begins feeding. Nearly all of the 23 terpenoids present exceeding 0.1% varied significantly either temporally or spatially, usually with complex interactions. Ordination and factor analysis revealed that terpenoids are less variable in mature leaf cushions than in young tissue. By entering a nonfeeding diapause during the late spring and summer, HWA avoids the unstable, variable levels of terpenoids in the immature leaf cushion and needles. PMID- 17078001 TI - Effect of a specific cyclooxygenase-gene polymorphism (A-842G/C50T) on the occurrence of peptic ulcer hemorrhage. AB - Cyclooxygenases (COX) catalyze the conversion of arachidonic acid to prostaglandins (PGs). COX-inhibiting drugs, such as nonsteroidal anti inflammatory drugs (NSAIDs), increase the risk for peptic ulcer disease. As a corollary, COX gene polymorphisms could be important in the pathogenesis of peptic ulcer disease because these affect prostaglandin formation and impair its protective effect at the level of the gastric mucosa. This study was designed to investigate the association between the functional single-nucleotide polymorphism, A-842G/C50T, in the COX-1 gene and peptic ulcer bleeding. We obtained DNA samples from 106 patients who underwent upper gastrointestinal endoscopy because of bleeding peptic ulcer disease and from 88 healthy control subjects. Genetic polymorphism in A-842G/C50T was determined by PCR followed by restriction-fragment-length-polymorphism analysis. Adjusted logistic regression analysis was performed to evaluate the associations. Risk factors associated with peptic ulcer bleeding were male gender (odds ratio, 4.78; 95% confidence interval, 2.6-8.8) and NSAID/aspirin-use (odds ratio, 38.39; 95% confidence interval, 14.2-103.6). The A-842G/C50T heterozygote was less frequent in peptic ulcer bleeding (n = 7) compared with healthy control subjects (n = 11). The adjusted risk for peptic ulcer bleeding among individuals who were heterozygote for the A-842G/C50T polymorphism was 0.75 (range, 0.19-3.01) compared with wild type. The COX-1 A-842G/C50T SNP does not influence the risk for developing peptic ulcer bleeding. PMID- 17078002 TI - Low intestinal glutamine level and low glutaminase activity in Crohn's disease: a rational for glutamine supplementation? AB - Intestinal glutamine utilization is integral to mucosal regeneration. We analyzed the systemic and intestinal glutamine status in Crohn's disease (CD) and evaluated the therapeutic effect of glutamine supplementation in an animal model of ileitis. In CD, glutamine concentrations were decreased systemically and in noninflamed and inflamed ileal/colonic mucosa. Mucosal glutaminase activities were depressed in the ileum independent of inflammation but were not different from controls in the colon. In experimental ileitis, oral glutamine feeding prevented macroscopic inflammation, enhanced ileal and colonic glutaminase activities above controls, and normalized the intestinal glutathione redox status. However, glutamine supplementation enhanced myeloperoxidase activity along the gastrointestinal tract and potentiated lipid peroxidation in the colon. In conclusion, glutamine metabolism is impaired in CD. In experimental ileitis, glutamine supplementation prevents inflammatory tissue damage. In the colon, however, which does not use glutamine as its principal energy source, immune enhancement of inflammatory cells by glutamine increases oxidative tissue injury. PMID- 17078003 TI - Involvement of cyclooxygenase-2--prostaglandin E2 pathway in interleukin-8 production in gastric cancer cells. AB - Prostaglandin E(2) (PGE(2)) is thought to play an important role in both inflammatory and anti-inflammatory effects. The effect of PGE(2) on the proinflammatory chemokine interleukin-8 (IL-8) in the gastric epithelial cells has not been defined yet. A gastric cancer cell line (MKN45) and primary gastric fibroblasts were cocultured with Helicobacter pylori standard strain (NCTC11637). The expressions of IL-8 and cyclooxygenase 2 (COX-2) mRNA were examined by reverse transcription polymerase chain reaction (RT-PCR) amplification. The amount of IL-8 antigen secreted by the MKN45 cells and gastric fibroblasts was measured by enzyme-linked immunosorbent assay (ELISA). We examined the effects of H pylori stimulation on IL-8 and COX-2 expression levels and the effects of COX-2 inhibitor on H pylori-induced IL-8 production in the MKN45 cells and gastric fibroblasts. Furthermore, we examined the expressions of subtypes of PGE(2) receptors, the effects of arachidonic acid and PGE(2) on IL-8 production, and the effects of PGE(2) on the total cellular cyclic adenosine monophosphate (cAMP) in MKN45 cells. MKN45 cells and gastric fibroblasts expressed IL-8 and COX-2 mRNA under stimulation with H pylori. The MKN45 cells produced IL-8 and PGE(2) antigen into the culture medium with H pylori stimulation, and the production level of IL 8 and PGE(2) antigen decreased significantly with COX-2 inhibitor pretreatment (concentration: 50 muM). On the other hand, the gastric fibroblasts strongly produced IL-8 antigen even in the unstimulated condition, and the amount of IL-8 antigen was not affected by H pylori stimulation and/or COX-2 inhibitor pretreatment. The MKN45 cells expressed IL-8 mRNA and released IL-8 antigen slightly, and the expression level of IL-8 mRNA and the amount of IL-8 antigen increased significantly with PGE(2) treatment in a dose-dependent manner. PGE(2) induced IL-8 production was inhibited by pretreatment with EP2 and EP4 antagonists. The MKN45 cells expressed EP2 and EP4 subtypes of PGE(2) receptors, and these expression levels were not affected by H pylori stimulation or PGE(2) treatment. The amount of IL-8 antigen increased slightly, but not significantly, with arachidonic acid treatment. PGE(2) treatment for 15 minutes increased the total cellular cAMP in the MKN45 cells. These results suggest that the COX-2 PGE(2) pathway may be involved in IL-8 production in gastric epithelial cells. PMID- 17078004 TI - Usefulness of the immunological rapid urease test for detection of Helicobacter pylori in patients who are reluctant to undergo endoscopic biopsies. AB - Cardiovascular diseases and liver cirrhosis are in the list of Helicobacter pylori-related extragastric diseases. Patients with cirrhosis and cardiovascular diseases under the control of aspirin have increased risk of gastrointestinal bleeding. The immunological rapid urease test (IRUT) provides a rapid and safe diagnostic test for H. pylori using gastric mucus collected at endoscopy. We investigated its usefulness in 93 patients with these extragastric diseases including 46 with H. pylori infection assessed by urea breath test and serology. Twenty H. pylori-infected patients received eradication therapy and the IRUT was assessed to evaluate the efficacy of bacterial eradication. The sensitivity, specificity, and positive and negative predictive values for IRUT were 96%, 90%, 90%, and 96%, respectively. The results of IRUT completely agreed with those of urea breath test following anti-H. pylori therapy. The IRUT has acceptable diagnostic performance in such cohorts reluctant to undergo endoscopic biopsies due to the risk of bleeding. PMID- 17078005 TI - Susceptibility-guided vs. empiric retreatment of Helicobacter pylori infection after treatment failure. AB - Successful eradication of Helicobacter pylori after failure of standard triple therapy is difficult because of the higher resistance to metronidazole and clarithromycin. We evaluated the efficacy of susceptibility-guided vs. empiric retreatment for H. pylori after at least one treatment failure and determined the prevalence of posttreatment antibiotic resistance. Forty-nine patients in whom at least one treatment regimen for H. pylori eradication had failed underwent gastric biopsy and culture and were retreated according to the in vitro susceptibility results. Findings were compared with those for 49 control patients referred to our center for a (13)C-urea breath test. H. pylori eradication was assessed by urea breath test at least 6 weeks after retreatment in both groups. Susceptibility-guided retreatment was associated with better eradication rates than empiric treatment. The difference remained significant in stratified and multivariate analysis. Susceptibility-guided retreatment appears to be significantly more effective than empiric retreatment in eradicating H. pylori after at least one previous treatment failure. PMID- 17078006 TI - Efficacy of probiotics and nutrients in functional gastrointestinal disorders: a preliminary clinical trial. AB - The purpose of this study was to evaluate the efficacy and safety of commonly used probiotics and nutrients available for functional gastrointestinal disorders (FGID). Five different combinations of probiotics and nutrients, or placebo, were taken daily over 12 weeks. In this randomized controlled clinical trial, men and women 21 to 72 years of age with FGID symptoms of indigestion, bloating, and abdominal discomfort were assigned to one of six groups, 12 patients per group. Gastrointestinal Quality of Life Index (GIQLI) and visual analogue scale (VAS) for GI symptoms, SF-36, lactulose and mannitol test (LMT), and urine indican levels were evaluated. GIQLI, VAS scores, and LMT did not change significantly (P > 0.05). There were clinically notable improvements in two of the combination formulations. While the nonsignificant improvements in GI symptoms could suggest that combination probiotics and nutrients may be beneficial in conditions such as FGID, no conclusive evidence was found in this pilot trial. Further investigations to explore the findings are discussed. PMID- 17078007 TI - Helicobacter pylori and Clostridium difficile in cystic fibrosis patients. AB - We describe the prevalence of H. pylori and toxigenic Clostridium difficile (CD) infection and its relationship with gastrointestinal symptoms and pancreatic sufficiency (PS) or insufficiency (PI) in cystic fibrosis (CF) patients. Stool specimens from 30 consecutive patients with CF, aged 1-44, and from 30 healthy similarly aged subjects were tested for the H. pylori antigen by specific monoclonal antibodies and for CD toxins by Tox A/B assay and Tox A assay. CF patients were assessed clinically and tested for specific H. pylori serum antibodies and for mutations. In CF patients, the prevalence of H. pylori antigen was 16.6% (5/30), compared to 30% (9/30) in controls. Of the 26 CF patients with PI, only 2 (7.6%) were infected by H. pylori, compared with 3 of the 4 (75%) patients with PS (P=0.001). H. pylori infection was diagnosed in 3 of 5 (60%) CF patients carrying mild mutations, compared to 1 of 25 (4%) CF patients carrying severe mutations (P=0.01). Fourteen of 30 (46.6%) stool specimens from CF patients tested positive in the ToxA/B assay, and 3 of 14 tested positive for ToxA. No significant differences in antibiotic use, severity of lung disease, PI, chronic abdominal pain, or genotype were found between the two groups. None of the controls was positive for CD toxins. Prevalence of H. pylori infection in CF patients was lower than in similarly aged non-CF controls. CF patients with PI or a history of distal intestinal obstruction syndrome and those carrying mutations associated with a severe phenotype were protected against H. pylori infection. Almost half of the CF patients were asymptomatic carriers of CD producing mostly toxin B. More studies are needed to confirm our results in a larger group of CF patients. PMID- 17078008 TI - High rate of percutaneous endoscopic gastrostomy site infections due to oropharyngeal colonization. AB - We prospectively analyzed peristomal infection agents during the first week following percutaneous endoscopic gastrostomy (PEG) placement by the pull technique in patients with head and neck cancer. Nasal and pharyngeal swabs were obtained from a consecutive series of cancer patients prior to PEG placement. All patients underwent antibiotic prophylaxis with cefotaxime and oral disinfection. PEG site infection was prospectively evaluated at days 2, 3, and 7 after insertion. Twenty-eight patients (25 males; mean age, 58 years) were included. Oropharyngeal or nasal colonization were observed in 68% (19/28) and 19% (5/28) of patients, respectively. Early infections occurred in 36% (10/28) of the patients, all with oropharyngeal colonization and concordance between agents (P=0.01). Three patients required hospital admission and 1 required surgery. Head and neck cancer patients with oropharyngeal colonization have a high risk of early PEG site infection with substantial morbidity owing to oral-stomal spread. PMID- 17078009 TI - Gastric electrical stimulation with parameters for gastroparesis enhances gastric accommodation and alleviates distention-induced symptoms in dogs. AB - Gastric electrical stimulation (GES) improves symptoms in patients with gastroparesis. We sought to determine if stimulation at fundus with parameters used for gastroparesis could affect gastric accommodation and distention-induced symptoms in dogs. Nine dogs were implanted with a gastric cannula at the anterior stomach and 1 pair of stimulation electrodes in the fundus. Assessment of gastric accommodation and a series of gastric distention were performed using a barostat. Stimulation parameters were of short pulse trains of 14 Hz, 5 mA, 0.3 ms, and 0.1 s on, 5 s off. GES at fundus significantly decreased fasting gastric tone. Fasting gastric volume was significantly increased from 56.3+/-10.4 mL at baseline to 102.4+/-23.1 mL with stimulation (P=.011). Postprandial gastric accommodation was significantly enhanced with stimulation. The extent of accommodation increased from 249.3+/-39.9 mL in the control session to 325.8+/ 25.1 mL with stimulation (P=.011). Symptom scores induced by balloon distention of the stomach were significantly lower during stimulation in comparison with those of baseline (P=.016). In conclusion, GES with parameters for gastroparesis enhances postprandial gastric accommodation and reduces visceral perception in normal dogs. This effect, if seen also in humans, may explain in part the symptomatic improvement associated with GES therapy in patients with gastroparesis. PMID- 17078010 TI - Significance of endoscopy in asymptomatic premenopausal women with iron deficiency anemia. AB - There is, currently, no consensus with regard to the role of endoscopy in the etiologic investigation of asymptomatic pemenopausal women suffering from iron deficiency anemia (IDA). We conducted a retrospective case-control study to evaluate the contribution of esophagogastroduodenoscopy (EGD) and colonoscopy to the etiologic diagnosis of a group of asymptomatic premenopausal women suffering from IDA. One hundred eight consecutive asymptomatic premenopausal women who fulfilled our entry criteria were included in our patient group between January 1998 and December 2004. One hundred thirty-five age-matched asymptomatic premenopausal women without anemia who had undergone EGD and colonoscopy for medical checkups were included in the control group. Clinically relevant lesions were detected in 7 of 108 (6.5%) of the patients and in 8 of 135 (5.9%) of the controls. There were no differences with regard to the frequency of clinically relevant lesions between the two groups (P > 0.05). Concomitant upper and lower GI lesions were not detected in any patients. In the upper GI tract, the only lesion found to be potentially causative of IDA anemia was a severe erosive gastritis, which was found in both the patient and the control groups. A source consistent with chronic bleeding was detected in the lower GI tract in 6 (5.6%) of the patients and 7 (5.2%) of the controls. Bleeding hemorrhoids represented the most frequently detected lesions in both the patient and control groups. Only one case of colon cancer was detected in the patient group. As IDA in the premenopausal women could not be attributed consistently to GI blood loss in this study, prospective studies should be conducted to validate our findings and to identify which subgroup of asymptomatic premenopausal women would benefit from a diagnostic endoscopic evaluation. PMID- 17078011 TI - Attitudes about stimulant medication for attention-deficit/hyperactivity disorder among African American families in an inner city community. AB - Limited information exists on views among African American families living in low income, inner-city communities regarding the treatment of children with attention deficit/hyperactivity disorder (ADHD). Parents of children treated for ADHD in an urban primary care setting were recruited to complete a survey to assess attitudes toward stimulant medications. Although most (71%) were initially hesitant to use stimulants based on what they heard in the lay press, 63% would recommend stimulant medication to a relative/friend whose child had ADHD. Approximately 17% believed stimulants led to drug abuse, 21% preferred counseling over medication, 21% felt medications had bad side effects, and 23% believed that too many children were medicated for ADHD. Most (90%) felt the medication was safe if a physician recommended it. Views did not differ between participants whose child had or had not received counseling. Additional studies are needed to clarify whether such views impact treatment choices and health outcomes. PMID- 17078012 TI - Validation of a real-time PCR for the quantitative estimation of a G143A mutation in the cytochrome bc1 gene of Pyrenophora teres. AB - A single nucleotide polymorphism (SNP) in the cytochrome b gene confers resistance to strobilurin fungicides for several fungal pathogens. Therefore, on the basis of a change at amino acid position 143 from glycine to alanine, a real time PCR assay was established for the quantitative detection of the analogous SNP in the cytochrome b sequence of Pyrenophora teres Drechsler, which causes barley net blotch. Allelic discrimination was achieved by using allele specific primers with artificially mismatched nucleic acid bases and minor groove binding probes. Validation parameters for the lower limits of the working range, namely limits of detection (LOD) and limits of quantification (LOQ), were statistically determined by the variance of calibration data, as well as by the variance of the 100% non-strobilurin-resistant allele DNA sample (blank values). It was found that the detection was limited by the variance of blank values (five in 801 458 copies; 0.0006%), whereas the quantification was limited by the variance of calibration data (37 in 801 458 copies; 0.0046%). The real-time PCR assay was finally used to monitor strobilurin-resistant cytochrome b alleles in barley net blotch field samples, which were already classified in in vivo biotests to be fully sensitive to strobilurins. All signals for strobilurin-resistant cytochrome b alleles were below the LOD, and therefore the results are in total agreement with the phenotypes revealed by biotests. PMID- 17078013 TI - Synthesis and fungicidal activity of N-2-(3-methoxy-4-propargyloxy) phenethyl amides. Part 3: stretched and heterocyclic mandelamide oomyceticides. AB - Novel analogues of mandipropamid have been designed and prepared. The synthetic approach to these stretched and heterocyclic mandelamides is outlined. Biological data demonstrate their high efficacy against important plant diseases like tomato and potato late blight (Phytophthora infestans De Bary) and grape downy mildew (Plasmopara viticola Berliner & de Toni). Structure-activity relationship studies are discussed. PMID- 17078014 TI - Development of a pheromone trap monitoring system for orange wheat blossom midge, Sitodiplosis mosellana, in the UK. AB - Field-trapping experiments with synthetic 2,7-nonadiyl dibutyrate, the female produced sex pheromone of the orange wheat blossom midge, Sitodiplosis mosellana (Gehin), demonstrated that pheromone traps were highly attractive to males and caught very few non-target organisms. Different formulations of pheromone were tested to identify the optimum release rate and dispenser type for use in pheromone traps in the UK. Key findings were that racemic pheromone was as effective as enantiomerically pure (2S,7R)-2,7-nonadiyl dibutyrate, that release rates higher than 0.5 microg day(-1) were not necessary and that the optimal formulation was a 1 mg pheromone loading in a rubber septum. Pheromone traps gave a reliable indication of peak midge emergence, onset of flight and abundance of midges throughout the season. A strong correlation between maximum trap catch and crop infestation levels was obtained. PMID- 17078017 TI - Proteomic analysis of cerebrospinal fluid discriminates malignant and nonmalignant disease of the central nervous system and identifies specific protein markers. AB - CNS diseases are often accompanied by changes in the protein composition of cerebrospinal fluid (CSF). SELDI-TOF-MS provides an approach for identifying specific protein markers of disease in biological fluids. We compared the CSF proteomes from patients with neoplastic and reactive/inflammatory CNS diseases to identify potential biomarkers. SELDI-TOF-MS was performed on CSF derived from lumbar puncture of 32 patients, including 10 with CNS malignancies, 12 with inflammatory or reactive conditions, and 10 with unknown CNS disease. Using the SAX-2 (strong anionic exchange) chip, we uncovered three conserved protein peak ranges within each disease category. For neoplastic diseases, we identified conserved peaks at 7.5-8.0 kDa (9/10 samples), 15.1-15.9 kDa (8/10 samples), and 30.0-32.0 kDa (5/10 samples). In reactive/inflammatory diseases, conserved peaks were found at 6.7-7.1 kDa (10/12 samples), 11.5-11.9 kDa (12/12 samples), and 13.3-13.7 kDa (9/12 samples). A protein from the 30.0 to 32.0 kDa peak range found in neoplastic CSF was identified by MALDI analysis as carbonic anhydrase, a protein overexpressed in many malignancies including high-grade gliomas. Similarly, cystatin C was identified in the 13.3-13.7 kDa peak range in non neoplastic CSF and was most prominent in inflammatory conditions. Our approach provides a rational basis for identifying biomarkers that could be used for detection, diagnosis, and monitoring of CNS diseases. PMID- 17078018 TI - Mass spectrometric genomic data mining: Novel insights into bioenergetic pathways in Chlamydomonas reinhardtii. AB - A new high-throughput computational strategy was established that improves genomic data mining from MS experiments. The MS/MS data were analyzed by the SEQUEST search algorithm and a combination of de novo amino acid sequencing in conjunction with an error-tolerant database search tool, operating on a 256 processor computer cluster. The error-tolerant search tool, previously established as GenomicPeptideFinder (GPF), enables detection of intron-split and/or alternatively spliced peptides from MS/MS data when deduced from genomic DNA. Isolated thylakoid membranes from the eukaryotic green alga Chlamydomonas reinhardtii were separated by 1-D SDS gel electrophoresis, protein bands were excised from the gel, digested in-gel with trypsin and analyzed by coupling nano flow LC with MS/MS. The concerted action of SEQUEST and GPF allowed identification of 2622 distinct peptides. In total 448 peptides were identified by GPF analysis alone, including 98 intron-split peptides, resulting in the identification of novel proteins, improved annotation of gene models, and evidence of alternative splicing. PMID- 17078019 TI - Proteomic analysis of glutathione transferases from the liver fluke parasite, Fasciola hepatica. AB - The parasite Fasciola hepatica causes major global disease of livestock, with increasing reports of human infection. Vaccine candidates with varying protection rates have been identified by pre-genomic approaches. As many candidates are part of protein superfamilies, sub-proteomics offers new possibilities to systematically reveal the relative importance of individual family proteins to vaccine formulations within populations. The superfamily glutathione transferase (GST) from liver fluke has phase II detoxification and housekeeping roles, and has been shown to contain protective vaccine candidates. GST were purified from cytosolic fractions of adult flukes using glutathione- and S-hexylglutathione agarose, separated by 2-DE, and identified by MS/MS, with the support of a liver fluke EST database. All previously described F. hepatica GST isoforms were identified in 2-DE. Amongst the isoforms mapped by 2-DE, a new GST, closely related to the Sigma class enzymes is described for the first time in the liver fluke. We also describe cDNA encoding putative Omega class GST in F. hepatica. PMID- 17078020 TI - Long standing priapism as presentation of lymphoma. PMID- 17078021 TI - Cognitive dysfunction in beta-thalassemia minor. AB - Beta-thalassemia minor is a common, hereditary, and mostly symptomless disease. Previous studies have shown that lower hemoglobin values are associated with poorer cognitive functions. We aimed to evaluate the cognitive function in patients with beta-thalassemia minor. Thirty-two male subjects with beta thalassemia minor and 32 sex-, age-, and education status-matched healthy subjects were enrolled in the study. Blood tests and P300 potentials were carried out. P300 potential latency in all patients was significantly longer than those in the control group (337.63+/-34.89 msec and 310.66+/-14.30 msec, respectively; t 4.046, P<0.001). The amplitudes of P300 in patient group were significantly lower than those in the control group (5.19+/-3.59 microV and 9.81+/-3.33 microV, respectively; t 5.349, P<0.001). In this study, we have found that P300 potentials are adversely affected for cognitive functions in patients with beta thalassemia minor. PMID- 17078022 TI - Consortium for osteogenesis imperfecta mutations in the helical domain of type I collagen: regions rich in lethal mutations align with collagen binding sites for integrins and proteoglycans. AB - Osteogenesis imperfecta (OI) is a generalized disorder of connective tissue characterized by fragile bones and easy susceptibility to fracture. Most cases of OI are caused by mutations in type I collagen. We have identified and assembled structural mutations in type I collagen genes (COL1A1 and COL1A2, encoding the proalpha1(I) and proalpha2(I) chains, respectively) that result in OI. Quantitative defects causing type I OI were not included. Of these 832 independent mutations, 682 result in substitution for glycine residues in the triple helical domain of the encoded protein and 150 alter splice sites. Distinct genotype-phenotype relationships emerge for each chain. One-third of the mutations that result in glycine substitutions in alpha1(I) are lethal, especially when the substituting residues are charged or have a branched side chain. Substitutions in the first 200 residues are nonlethal and have variable outcome thereafter, unrelated to folding or helix stability domains. Two exclusively lethal regions (helix positions 691-823 and 910-964) align with major ligand binding regions (MLBRs), suggesting crucial interactions of collagen monomers or fibrils with integrins, matrix metalloproteinases (MMPs), fibronectin, and cartilage oligomeric matrix protein (COMP). Mutations in COL1A2 are predominantly nonlethal (80%). Lethal substitutions are located in eight regularly spaced clusters along the chain, supporting a regional model. The lethal regions align with proteoglycan binding sites along the fibril, suggesting a role in fibril-matrix interactions. Recurrences at the same site in alpha2(I) are generally concordant for outcome, unlike alpha1(I). Splice site mutations comprise 20% of helical mutations identified in OI patients, and may lead to exon skipping, intron inclusion, or the activation of cryptic splice sites. Splice site mutations in COL1A1 are rarely lethal; they often lead to frameshifts and the mild type I phenotype. In alpha2(I), lethal exon skipping events are located in the carboxyl half of the chain. Our data on genotype-phenotype relationships indicate that the two collagen chains play very different roles in matrix integrity and that phenotype depends on intracellular and extracellular events. PMID- 17078023 TI - Transformation of adult retina from the regenerative to the axonogenesis state activates specific genes in various subsets of neurons and glial cells. AB - The purpose of this study was to identify the gene expression profile of the regenerating retina in vitro. To achieve this goal, three experimental groups were studied: (1) an injury control group (OC-LI group) that underwent open crush (OC) of the optic nerve and lens injury (LI) in vivo; (2) an experimental group (OC-LI-R group) that comprised animals treated like those in the OC-LI group except that retinal axons were allowed to regenerate (R) in vitro; and (3) an experimental group (OC-LI-NR group) that comprised animals treated as those in the OC-LI group, except that the retinas were cultured in vitro with the retinal ganglion cell (RGC) layer facing upwards to prevent axonal regeneration (NR). Gene expression in each treatment group was compared to that of untreated controls. Immunohistochemistry was used to examine whether expression of differentially regulated genes also occurred at the protein level and to localize these proteins to the respective retinal cells. Genes that were regulated belonged to different functional categories such as antioxidants, antiapoptotic molecules, transcription factors, secreted signaling molecules, inflammation related genes, and others. Comparison of changes in gene expression among the various treatment groups revealed a relatively small cohort of genes that was expressed in different subsets of cells only in the OC-LI-R group; these genes can be considered to be regeneration-specific. Our findings demonstrate that axonal regeneration of RGC involves an orchestrated response of all retinal neurons and glia, and could provide a platform for the development of therapeutic strategies for the regeneration of injured ganglion cells. PMID- 17078024 TI - Upregulation of CCAAT/enhancer binding protein beta in activated astrocytes and microglia. AB - The transcription factor CCAAT/enhancer binding protein beta (C/EBPbeta) regulates the expression of key genes in inflammation but little is known about the involvement of C/EBPbeta in glial activation. In this report, we have studied the patterns of astroglial and microglial C/EBPbeta expression in primary mouse cortical cultures. We show that both astrocytes and microglia express C/EBPbeta in untreated mixed glial cultures. C/EBPbeta is upregulated when glial activation is induced by lipopolysaccharide (LPS). The LPS-induced upregulation of glial C/EBPbeta is rapid (2 h at mRNA level, 4 h at protein level). It is elicited by low concentrations of LPS (almost maximal effect at 1 ng/mL) and it is reversed by the protein synthesis inhibitor cycloheximide. C/EBPbeta nuclear levels increase both in astrocytes and microglia after LPS treatment, and the response is more marked in microglia. The LPS-induced increase in microglial C/EBPbeta is prevented by coadministration of the MAP kinase inhibitors SB203580 (p38 inhibitor) + SP600125 (JNK inhibitor) or SB203580 + U0126 (ERK inhibitor). Systemic injection of LPS also increases brain nuclear levels of C/EBPbeta as shown by Western blot, and this increase is localized in microglial cells as shown by double immunofluorescence, in the first report to our knowledge of C/EBPbeta expression in activated glial cells in vivo. These findings support a role for C/EBPbeta in the activation of astrocytes and, particularly, microglia. Given the nature of the C/EBPbeta-regulated genes, we hypothesize that this factor participates in neurotoxic effects associated with glial activation. (c) 2006 Wiley-Liss, Inc. PMID- 17078025 TI - Intima-media thickness of the common carotid artery in highway toll collectors. AB - PURPOSE: To assess the effects of exposure to exhaust particles on intima-media thickness of the common carotid artery in highway toll collectors. METHODS: Sixty one highway toll collectors (HTCs) between 24 and 56 years of age (mean, 36.2 +/- 7.3) and 48 controls between 24 and 64 years of age (mean, 42.6 +/- 10.6) were evaluated with gray-scale sonography to measure intima-media thickness (IMT) of the common carotid artery (CCA). Subgroups were categorized according to duration of exhaust exposure and further divided according to tobacco use. RESULTS: CCA IMT was higher (0.8 +/- 0.2 mm) in HTCs than in the control group (0.6 +/- 0.1 mm; p < 0.001) and remained higher when subgroups with similar smoking habits were compared. In HTCs, IMT was greater when the number of years working in tollbooths was greater (p = 0.023). IMT was lower in HTCs with an exposure duration of < 10 years compared with a duration of 10-20 years (p = 0.017) or > 20 years (p value not significant). CONCLUSION: Air pollution has a widely acknowledged negative effect on humans. This study confirms that exposure to exhaust particles might cause wall thickening of carotid arteries. PMID- 17078026 TI - S100B expression defines a state in which GFAP-expressing cells lose their neural stem cell potential and acquire a more mature developmental stage. AB - During the postnatal development, astrocytic cells in the neocortex progressively lose their neural stem cell (NSC) potential, whereas this peculiar attribute is preserved in the adult subventricular zone (SVZ). To understand this fundamental difference, many reports suggest that adult subventricular GFAP-expressing cells might be maintained in immature developmental stage. Here, we show that S100B, a marker of glial cells, is absent from GFAP-expressing cells of the SVZ and that its onset of expression characterizes a terminal maturation stage of cortical astrocytic cells. Nevertheless, when cultured in vitro, SVZ astrocytic cells developed as S100B expressing cells, as do cortical astrocytic cells, suggesting that SVZ microenvironment represses S100B expression. Using transgenic s100b-EGFP cells, we then demonstrated that S100B expression coincides with the loss of neurosphere forming abilities of GFAP expressing cells. By doing grafting experiments with cells derived from beta-actin-GFP mice, we next found that S100B expression in astrocytic cells is repressed in the SVZ, but not in the striatal parenchyma. Furthermore, we showed that treatment with epidermal growth factor represses S100B expression in GFAP-expressing cells in vitro as well as in vivo. Altogether, our results indicate that the S100B expression defines a late developmental stage after which GFAP-expressing cells lose their NSC potential and suggest that S100B expression is repressed by adult SVZ microenvironment. PMID- 17078027 TI - Effects of inflammatory cytokines IL-1beta, IL-6, and TNFalpha on the intracellular localization of retinoid receptors in Schwann cells. AB - It was investigated whether retinoic acid (RA) and the proinflammatory cytokines IL-1beta, IL-6, and TNFalpha influence the intracellular distribution of retinoic acid receptors (RAR) and retinoid X receptors (RXR) in Schwann cells. This question arose because nuclear translocation of RARalpha, RXRalpha, and RXRbeta was observed after nerve injury, and because mutual interactions exist between the signal transduction pathways of RA and proinflammatory cytokines. Schwann cell primary cultures from the rat sciatic nerve were incubated with IL-1beta, IL 6, and TNFalpha, with all-trans RA and with a combination of IL-1beta and RA. After incubation periods ranging from 5 min to 5 h, the intracellular distributions of RARalpha, RARbeta, RXRalpha, and RXRbeta were analyzed. All three cytokines caused a shift of RARalpha from the cytosolic compartments into the cell nuclei. This was also observed with RA, and combining RA with IL-1beta produced an additive effect. IL-1beta and IL-6 also affected the distribution of RARbeta, although immunoreactivity of this receptor always remained stronger in the cytosol. No effect of the cytokines on RXRalpha or RXRbeta was observed, whereas RA treatment caused a stronger nuclear signal of both receptors. Effects on the subcellular localization of retinoid receptors may provide a link in a feedback loop between RA/RAR and cytokines. PMID- 17078028 TI - Calponin is expressed by fibroblasts and meningeal cells but not olfactory ensheathing cells in the adult peripheral olfactory system. AB - Olfactory ensheathing cells (OECs), the principal glial cells of the peripheral olfactory system, have many phenotypic similarities with Schwann cells of the peripheral nervous system. This makes reliably distinguishing these two cells types difficult, especially following transplantation into areas of injury in the central nervous system. In an attempt to identify markers by which these two cells types can be distinguished, a recent proteomic analysis of fetal OECs and adult Schwann cells identified the actin-binding protein calponin as a potential marker expressed by OECs but not Schwann cells. Since many studies designed with the translational goal of autologous transplantation in mind have used adult OECs, this study examined the expression of calponin by adult OECs, both in vivo within the peripheral olfactory system and in vitro. Calponin colocalized with strongly fibronectin positive fibroblasts in the olfactory mucosa (OM) and meningeal cells in the olfactory bulb (OB) but not with S100beta or neuropeptide Y positive OECs. In tissue culture, calponin was strongly expressed by fibronectin-expressing fibroblasts from OM, sciatic nerve and skin and by meningeal cells from the OB, but not by p75(NTR)- and S100beta-expressing OECs. These data, supported by Western blotting, indicate that calponin can not be used to distinguish adult OECs and Schwann cells. PMID- 17078029 TI - Value of parathyroid hormone assay for preoperative sonographically guided parathyroid aspirates for minimally invasive parathyroidectomy. AB - PURPOSE: The key to successful parathyroid surgery is accurate preoperative tumor localization. This study investigates the use of ultrasound (US)-guided parathyroid fine needle aspiration (FNA) as a confirmatory diagnostic method in patients with hyperparathyroidism undergoing minimally invasive parathyroidectomy. METHODS: Patients were selected for minimally invasive parathyroidectomy based on the finding of a single parathyroid adenoma identified with US and/or sestamibi scans and confirmation of the suspected parathyroid lesion via FNA and parathyroid hormone (PTH) assay. The value of aspirate obtained from the thyroid gland intraoperatively served as the negative control. RESULTS: A total of 56 tissue FNAs were performed in 27 patients. US detected masses suggestive of parathyroid lesion in all 27 patients, and 31 US-guided FNAs were performed. No complications related to the procedure were noted. Intraoperatively, FNA was performed in the thyroids of 25 patients undergoing minimally invasive parathyroidectomy. Aspirates from lesions subsequently confirmed as having developed from the parathyroid gland had a mean PTH level of 4,677 +/- 123 pg/ml (range, 3,600-5,000 pg/ml), which was significantly higher than thyroid aspirates, which yielded a mean PTH level of 48 +/- 7 pg/ml (range, 5-57 pg/ml). The sensitivity of US and sestamibi scans in the detection of abnormal parathyroid glands was 88% and 77%, respectively. The sensitivity of US guided FNA in confirming the parathyroid origin of a lesion was 100%. CONCLUSION: US-guided FNA for PTH assay can be performed safely for the confirmation of lesions identified with preoperative US for the selection of patients eligible for minimally invasive parathyroidectomy. PMID- 17078030 TI - Simvastatin regulates oligodendroglial process dynamics and survival. AB - Simvastatin, a lipophilic statin that crosses the blood-brain barrier, is being evaluated as a potential therapy for multiple sclerosis (MS) due to its anti inflammatory properties. We assessed the effects of simvastatin on cultures of rat newborn and human fetal oligodendrocyte progenitor cells (OPCs) and human adult mature oligodendrocytes (OLGs) with respect to cellular events pertaining to myelin maintenance and repair. Short-term simvastatin treatment of OPCs (1 day) induced robust process extension, enhanced differentiation to a mature phenotype, and decreased spontaneous migration. These effects were reversed by isoprenoid products and mimicked with an inhibitor of Rho kinase (ROCK), the downstream effector of the isoprenylated protein RhoA GTPase. Prolonged treatment (2 days) caused process retraction that was rescued by cholesterol, and increased cell death (4 days) partially rescued by either cholesterol or isoprenoid co treatment. In comparison, simvastatin treatment of human mature OLGs required a longer initial time course (2 days) to induce significant process outgrowth, mimicked by inhibiting ROCK. Prolonged treatment of mature OLGs was associated with process retraction (6 days) and increased cell death (8 days). Human-derived OPCs and mature OLGs demonstrated an increased sensitivity to simvastatin relative to the rodent cells, responding to nanomolar versus micromolar concentrations. Our findings indicate the importance of considering the short- and long-term effects of systemic immunomodulatory therapies on neural cells affected by the MS disease process. (c) 2006 Wiley-Liss, Inc. PMID- 17078031 TI - Biological structure and health implications from tooth size at Mission San Luis de Apalachee. AB - This study analyzes dental metric variation to examine the biological structure of the native population at Mission San Luis de Apalachee, a late 17th century mission located in the Apalachee Province of Spanish colonial Florida. Three topics are addressed: (1) comparison of tooth sizes among adult and subadults, (2) analysis of the bio-spatial structure of skeletons within the church area, and (3) comparison of phenotypic profiles of individuals interred within coffins in the ritual nucleus of the church: the altar region. Analyses indicate that subadults had smaller average tooth sizes than adults for the posterior dentition that was particularly evident in mandibular nonpolar molars and premolars. This disparity, also documented in two other mission populations, likely represents ontogenetic stress and resulting increased mortality among those most at risk for early death. Analysis of the spatial structure of graves failed to document biological structuring by side of the aisle or by burial row, although some gross differences were evident when front, middle, and rear church burials were compared. Individuals buried in coffins within the same row were phenotypically similar to one another. However, inter-row comparisons indicated lack of phenotypic similarity among all coffin interments. These analyses suggest maintenance of kin-structured burial for elites alone within the San Luis community. PMID- 17078032 TI - Mineralization of the mandibular third molar: a study of American blacks and whites. AB - The tempo of tooth mineralization is under significant genetic control, and the orderly progression of morphological changes-in concert with the long span during growth in which teeth form-makes "dental age" a useful measure of a person's degree of biological maturity. The third molar is of particular interest because (1) it is the last and most variable tooth to form and (2) it is the only tooth to complete formation after puberty, which has made it attractive in forensic and legal circles as an estimator of adulthood. Age standards are described here for mandibular third molar formation stages in a cross-sectional sample of 4,010 persons (age range: 3-25 years), with proportionate sample sizes of American blacks and whites and males and females. Formation was scored against the 15 grade ordinal scheme of Moorrees, and descriptive statistics were computed using proportional hazards survival analysis. Blacks achieved each formation stage significantly ahead of whites, but not in a uniform manner. Instead, there was an enhanced advancement in blacks during crown formation and during late stages of root formation. In both races formation proceeded faster in males, which is unique for the third molar, as prior studies suggest. Sample variance increases with the stage of formation, such that 95% confidence limits span 8 or more years for root formation stages. Consequently, the third molar provides a rough gauge of an individual's chronological age, but the considerable variability precludes any precise estimate, particularly in late adolescence where most forensic interest has focused. PMID- 17078033 TI - An investigation of skeletal indicators of vitamin D deficiency in adults: effective markers for interpreting past living conditions and pollution levels in 18th and 19th century Birmingham, England. AB - Vitamin D deficiency in adults is referred to as osteomalacia, and this condition has multiple causes related to factors such as environment, living conditions and cultural practices. Therefore, understanding the types and range of expression of osteomalacia in archaeological bone, using a number of investigative techniques, will have significant implications for interpretations made about past communities. This study aimed to understand the expression of vitamin D deficiency in the skeletons of adults through detailed analysis of human bone from the late 18(th) and 19(th) century churchyard of St. Martin's, Birmingham, England, at a gross, radiological and histological level. The collection from St. Martin's is unusual for the period as this central burial ground in Birmingham was used by a wide cross section of society rather than a narrow socio-economic group as at other sites of this date. Macroscopic and radiological analysis of 291 adults identified seven individuals with osteomalacia, and histological analysis using back-scattered scanning electron microscopy confirmed the findings. Detailed description of the range of pathological alterations observed in the archaeological skeletons are presented, and possible interpretations of the patterns seen considered. The results of this investigation will enable clear diagnosis and interpretation of this vitamin D deficiency disease, an important socio-economic indicator, to take place in the future. PMID- 17078034 TI - An elusive paleodemography? A comparison of two methods for estimating the adult age distribution of deaths at late Classic Copan, Honduras. AB - Comparison of different adult age estimation methods on the same skeletal sample with unknown ages could forward paleodemographic inference, while researchers sort out various controversies. The original aging method for the auricular surface (Lovejoy et al., 1985a) assigned an age estimation based on several separate characteristics. Researchers have found this original method hard to apply. It is usually forgotten that before assigning an age, there was a seriation, an ordering of all available individuals from youngest to oldest. Thus, age estimation reflected the place of an individual within its sample. A recent article (Buckberry and Chamberlain, 2002) proposed a revised method that scores theses various characteristics into age stages, which can then be used with a Bayesian method to estimate an adult age distribution for the sample. Both methods were applied to the adult auricular surfaces of a Pre-Columbian Maya skeletal population from Copan, Honduras and resulted in age distributions with significant numbers of older adults. However, contrary to the usual paleodemographic distribution, one Bayesian estimation based on uniform prior probabilities yielded a population with 57% of the ages at death over 65, while another based on a high mortality life table still had 12% of the individuals aged over 75 years. The seriation method yielded an age distribution more similar to that known from preindustrial historical situations, without excessive longevity of adults. Paleodemography must still wrestle with its elusive goal of accurate adult age estimation from skeletons, a necessary base for demographic study of past populations. PMID- 17078035 TI - Y-chromosomal variation in the Czech Republic. AB - To analyze the contribution of the Czech population to the Y-chromosome diversity landscape of Europe and to reconstruct past demographic events, we typed 257 males from five locations for 21 UEPs. Moreover, 141 carriers of the three most common haplogroups were typed for 10 microsatellites and coalescent analyses applied. Sixteen Hg's characterized by derived alleles were identified, the most common being R1a-SRY(10831) and P-DYS257*(xR1a). The pool of haplogroups within I M170 represented the third most common clade. Overall, the degree of population structure was low. The ages for Hg I-M170, P-DYS257*(xR1a), and R1a-SRY(10831) ap peared to be comparable and compatible with their presence during or soon after the LGM. A signal of population growth beginning in the first millennium B.C. was detected. Its similarity among the three most common Hg's indicated that growth was characteristic for a gene pool that already contained all of them. The Czech population appears to be influenced, to a very moderate extent, by genetic inputs from outside Europe in the post-Neolithic and historical times. Population growth postdated the archaeologically documented introduction of Neolithic technology and the estimated central value coincides with a period of repeated changes driven by the development of metal technologies and the associated social and trade organization. PMID- 17078036 TI - Environmental effects on skeletal versus dental development: Using a documented subadult skeletal sample to test a basic assumption in human osteological research. AB - This study examines the relationship between measures of skeletal and dental development and socioeconomic factors in a 20th century documented skeletal sample of children from Portugal. The skeletons are of known sex and chronological age, and include other biographic data, such as cause of death. Growth in the length of the long bone is used as a measure of skeletal growth, and schedules of tooth formation are used as a measure of dental development. These two measures of physiological age were compared to chronological age, to assess growth and developmental status. Socioeconomic indicators were obtained from the supporting documentation, and include the occupation of the father and the place of residence, which were used to build a socioeconomic classification based on two groups, one of low and the other of high socioeconomic status. Growth and development status was then compared in these two groups. Results show that socioeconomic differences are much more pronounced in skeletal growth than in dental development. This largely supports the assertion that dental development is buffered against environmental factors relative to skeletal development. However, in this study, skeletal maturation could not be assessed, and findings indicate that dental development can show significant delays at the lower end of the socioeconomic gradient. PMID- 17078037 TI - Carabelli's trait and tooth size of human maxillary first molars. AB - Carabelli's trait is a morphological feature that can occur on the protocone of human maxillary molars. This study tests the hypothesis that Carabelli's trait is correlated statistically with the dimensions of the crown's four principal cusps or whether, as a cingular feature, the trait truly accretes onto an otherwise unaffected crown. Computer-assisted image analysis was used to measure the 6 intercusp distances and 12 angular relationships among cusp tips on the permanent first molar of 300 young adult American whites. Carabelli's complex was scored using an 8-grade ordinal scheme. Crown size was quantified in three ways, namely as 1) maximum mesiodistal and buccolingual diameters, 2) the 6 intercusp distances, and 3) the 12 angular cusp arrangements. There was no sex difference in the morphological expression of Carabelli's trait in this sample. Overall crown size and intercusp distances were significantly and progressively larger in molars with larger Carabelli's trait expressions. There are graded size responses between crown size (mesiodistal and buccolingual diameters), sizes of the four principal cusps, and morphological stage of Carabelli's complex, though the statistical relationships are appreciably stronger in males than females. Carabelli's trait occurs preferentially in larger molars. In contrast, angular (shape) relationships among cusp tips are not discernibly affected by trait size in either sex. There is the situation, then, that Carabelli's trait is developmentally correlated with crown size, but with no apparent alteration of cusp arrangements, suggesting that the increases are isometric across the occlusal table. Why the association is much weaker in females remains speculative, but these data provide yet another line of evidence that, within a population, tooth size is associated in a positive fashion with crown complexity. PMID- 17078038 TI - Inferring Plio-Pleistocene southern African biochronology from facial affinities in Parapapio and other fossil papionins. AB - Buried in the same South African cave deposits as Australopithecus, fossil papionins have been referred to Parapapio (Pp. whitei, Pp. broomi, Pp. jonesi, Pp. antiquus), Papio (P. izodi, P. angusticeps, P. h. robinsoni), Theropithecus (e.g., T. darti), Gorgopithecus, or Dinopithecus on the basis of postcanine tooth size and descriptive morphology of the muzzle. The morphological patterns of variation that these papionins demonstrate can help to place the Australopithecus fossils into a biochronological context and provide valuable information for reconstructing regional Plio-Pleistocene turnover. To document these patterns of variation across fossil-bearing sites, we explore morphometric affinities within Parapapio, and between Parapapio and other Plio-Pleistocene taxa (Dinopithecus ingens, Papio angusticeps, Papio izodi, and Theropithecus darti) by analyzing a sample of interlandmark distances derived from 3-D coordinate data of the most complete fossil papionin specimens available. Bivariate and multivariate analyses show that Pp. whitei exhibits as much variation between sites and between individuals as Pp. broomi and Pp. whitei combined. Diversity in Parapapio at Makapansgat and Sterkfontein may suggest substantial time depth to the caves. Theropithecus darti, Dinopithecus ingens, Papio angusticeps, Pp. whitei from Bolt's Farm (BF 43), and Pp. jonesi from Sterkfontein (STS 565) differ considerably from one another. Other Parapapio specimens across sites form a separate cluster with P. izodi from Taung, suggesting a Pliocene age for this site. PMID- 17078039 TI - Bioavailability and pharmacokinetic model for ritonavir in the rat. AB - The aim of this study is to investigate in vivo the oral bioavailability of ritonavir and to evaluate the pharmacokinetic model that best describes the plasma concentration behavior after oral and intravenous administration. Male Wistar rats were intravenously administered at 3 mg dose of pure ritonavir and oral administered at 4.6 +/- 2.5 mg of diluted Norvir. Blood samples were taken by means of the jugular vein for a 24 h period of time. An analytical high performance liquid chromatography (HPLC) technique was developed in order to quantify ritonavir plasma concentrations. A nonlinear modeling approach was used to estimate the pharmacokinetic parameters of interest. Results showed that a two compartmental model with zero-order kinetic in the incorporation process of ritonavir into the body better fitted intravenous and oral data. The estimated oral bioavailability by means of noncompartmental and compartmental approaches resulted in 74% and 76.4%, respectively. These values confirm the ones obtained by other authors in the rat. In conclusion, a zero-order kinetic in the incorporation process at the administered doses suggests the saturation of the possible specialized transport mechanisms involved in the incorporation of ritonavir into the body. These results could justify the use of low doses of ritonavir when improving the bioavailability of other protease inhibitors (PIs) is required. PMID- 17078040 TI - High-level expression of DNA architectural factor HMGA2 and its association with nucleosomes in human embryonic stem cells. AB - The state of chromatin in human embryonic stem (hES) cells is a key factor determining stem cell identity. The non-histone chromatin-associated factor HMGA2 has been studied mostly in the mouse where its function seems critical for embryonic cell growth and adipocytic cell differentiation. Here we show that HMGA2 is highly expressed in two undifferentiated human embryonic stem cell lines at a level of at least 10(5) copies per individual stem cell. Interestingly, expression is further upregulated by a factor of three at day 7 of embryoid body formation, before it quickly drops to or below the level found in undifferentiated cells. We also show that HMGA2 is stably associated with inter- and metaphase hES cell chromatin, and that up to 12 HMGA2 protomers stably associate in vitro with a single nucleosome core particle of known atomic structure. Our data lend support to the possibility that HMGA2 interacts with nucleosomes in a way that imposes a global effect on the state of ES cell chromatin, which may contribute to the establishment of both ES cell identity and the initiation of specific differentiation programs. PMID- 17078041 TI - 3D in silico modeling of the human respiratory system for inhaled drug delivery and imaging analysis. AB - The efficacies of inhaled pharmacologic drugs could be improved if drugs could be targeted to appropriate sites within the human respiratory system. The spatial deposition patterns of particles can now be detected with a high degree of resolution using advanced techniques of imaging (e.g., SPECT). However, the effectiveness of such laboratory regimens has been limited by the inability to clearly identify airway composition within images. Therefore, we have developed a theoretical protocol to map airways within human lungs that is designed to be used in a complementary manner with laboratory investigations. The in silico model has two components: a mathematical model based on concepts of topology; and, a computer algorithm which tracks the millions of constituent lung airways. The in silico model produces 3D lung structures that are anatomically correct and can be customized to each patient. We have applied the protocol to a SPECT study where the interiors of lungs were partitioned into a series of ten nested shells. Airway composition in the respective shells provides a heretofore unavailable quantification of scintigraphy images. The protocol can be employed in a practical manner in the medical arena to aid in the interpretation of SPECT images, and to provide a platform for the design of human subject tests. PMID- 17078042 TI - A gene trap knockout of the abundant sperm tail protein, outer dense fiber 2, results in preimplantation lethality. AB - Outer dense fiber 2 (Odf2) is highly expressed in the testis where it encodes a major component of the outer dense fibers of the sperm flagellum. Furthermore, ODF2 protein has recently been identified as a widespread centrosomal protein. While the expression of Odf2 highlighted a potential role for this gene in male germ cell development and centrosome function, the in vivo function of Odf2 was not known. We have generated Odf2 knockout mice using an Odf2 gene trapped embryonic stem cell (ESC) line. Insertion of a gene trap vector into exon 9 resulted in a gene that encodes a severely truncated protein lacking a large portion of its predicted coil forming domains as well as both leucine zipper motifs that are required for protein-protein interactions with ODF1, another major component of the outer dense fibers. Although wild-type and heterozygous mice were recovered, no mice homozygous for the Odf2 gene trap insertion were recovered in an extended breeding program. Furthermore, no homozygous embryos were found at the blastocyst stage of embryonic development, implying a critical pre-implantation role for Odf2. We show that Odf2 is expressed widely in adults and is also expressed in the blastocyst stage of preimplantation development. These findings are in contrast with early studies reporting Odf2 expression as testis specific and suggest that embryonic Odf2 expression plays a critical role during preimplantation development in mice. PMID- 17078043 TI - Stanley Fahn Lecture 2005: The staging procedure for the inclusion body pathology associated with sporadic Parkinson's disease reconsidered. AB - The synucleinopathy known as sporadic Parkinson's disease (PD) is a multisystem disorder that severely damages predisposed nerve cell types in circumscribed regions of the human nervous system. A recent staging procedure for the inclusion body pathology associated with PD proposes that, in the brain, the pathological process (formation of proteinaceous intraneuronal Lewy bodies and Lewy neurites) begins at two sites and continues in a topographically predictable sequence in six stages, during which components of the olfactory, autonomic, limbic, and somatomotor systems become progressively involved. In stages 1 to 2, the Lewy body pathology is confined to the medulla oblongata/pontine tegmentum and anterior olfactory structures. In stages 3 to 4, the substantia nigra and other nuclei of the basal mid- and forebrain become the focus of initially subtle and, then, severe changes. During this phase, the illness probably becomes clinically manifest. In the final stages 5 to 6, the lesions appear in the neocortex. This cross-sectional study originally was performed on 168 autopsy cases using material from 69 incidental cases and 41 clinically diagnosed PD patients as well as 58 age- and gender-matched controls. Here, the staging hypothesis is critically reconsidered and discussed. PMID- 17078044 TI - The anterior visceral endoderm-turning heads. AB - The Anterior Visceral Endoderm is an extraembryonic tissue that plays a pivotal role during embryogenesis, being responsible for the proper orientation of the anterior-posterior axis of the embryo and for appropriate pattering of adjacent embryonic tissue. In this review I discuss the formation and migration of the AVE, and attempt to place some recent findings in the context of a working model. PMID- 17078045 TI - Impaired long-term potentiation-like plasticity of the trigeminal blink reflex circuit in Parkinson's disease. AB - We investigated the hypothesis that Parkinsons's disease (PD) is associated with abnormal plasticity of the neuronal circuits mediating blink reflex. We induced long-term potentiation (LTP)-like plasticity in trigeminal wide dynamic range neurons of the blink reflex circuit by pairing an high-frequency train of electrical stimuli over the right supraorbital nerve (SO) coincident with the R2 response elicited by a preceding SO stimulus. The facilitation of the R2 response after the induction protocol was markedly decreased in patients relative to controls. Treatment with dopaminergic drugs normalized the LTP-like plasticity of the R2 response. We conclude that nigrostriatal denervation disrupts LTP-like plasticity in the trigeminal reflex circuit. PMID- 17078046 TI - Controlling diffusion of 3He by buffer gases: a structural contrast agent in lung MRI. AB - PURPOSE: To study the influence of admixing inert buffer gases to laser-polarized (3)He in terms of resulting diffusion coefficients and the consequences for image contrast and resolution. MATERIALS AND METHODS: The diffusion coefficient of (3)He was altered by admixing buffer gases of various molecular weights ((4)He, N(2), and SF(6)). The influence of the pulse sequence and the diffusion coefficient on the appearance of MRI of (laser-polarized) gases was analyzed by comparison of basic theoretical concepts with demonstrative experiments. RESULTS: Excellent agreement between theoretical description and observed signal in simple gradient echoes was observed. A maximum signal gain can be predicted and was experimentally validated. Images acquired under such conditions revealed improved resolution. The nature and concentration of the admixed gas defines a structural threshold for the observed apparent diffusion coefficient (ADC) as demonstrated with diffusion-weighted MRI on a pig's lung flooded with suitable gas mixtures. CONCLUSION: A novel procedure is proposed to control the diffusion coefficient of gases in MRI by admixture of inert buffer gases. Their molecular mass and concentration enter as additional parameters into the equations that describe structural contrast. This allows for setting a structural threshold up to which structures contribute to the image. For MRI of the lung this enables images of very small structural elements (alveoli) only, or in the other extreme, all airways can be displayed with minimal signal loss due to diffusion. PMID- 17078047 TI - Development of a three-dimensional finite element model of a human mandible containing endosseous dental implants. II. Variables affecting the predictive behavior of a finite element model of a human mandible. AB - The purpose of this study was to propose a systematic approach to validate a finite element model (FEM) of the human mandible and to investigate the effects of changing the geometry and orthotropic material properties on the FEM predictions. Thirty-eight variables affecting the material properties, boundary conditions, and the geometry of a FEM of a human mandible, including two dental implants, were systematically changed, creating a number of FEMs of the mandible. The effects of the variations were quantified as differences in the principal strain magnitudes modeled by the original FEM (gold standard), prior to the sensitivity analyses, and those generated by the changed FEMs. The material properties that had the biggest impact on the predicted cortical principal strain were the shear moduli (up to 31% in difference from the unchanged state), and the absence of cancellous bone (up to 34%). Alterations to the geometry of the mandibular cross section, such as an increase in corpus dimensions, had the greatest effect on principal strain magnitudes (up to 16%). Changes in the cortical thickness in relation to the width of the corpus section modified strain more than alterations to the corpus depth (14% and 5%, respectively). The relatively small difference (up to 13.5%) between the predicted and measured interimplant distances indicates the accuracy of the FEM. Changes in geometry and orthotropic material properties could induce significant changes in strain patterns. These values must therefore be chosen with care when using finite element techniques for predicting stresses, strains, and displacements. PMID- 17078048 TI - Development of a three-dimensional finite element model of a human mandible containing endosseous dental implants. I. Mathematical validation and experimental verification. AB - The purpose of this study was to mathematically validate and clinically verify a finite element model (FEM) of the human mandible and to compare the functional deformation predicted by the model with that detected clinically. Mandibular surface strain, using 11 single strain gauges, and medial convergence (MC), using a custom-fabricated linear variable differential transformer (LVDT), were recorded in a dried human jaw. The mandible was treated with two endosseous implants, placed bilaterally in the premolar area, and mounted in a rig that simulated natural function of the jaw. Measurements were made in real time using a multichannel analogue/digital converter and a personal computer for data storage and analysis. A FEM of the mandible was constructed, using commercially available finite element software, based on CT scan images of the jaw. MC was predicted as the linear change in the orientation of the two implants in the horizontal plane. Predicted and measured values of MC ranged between 60 and 109 microm. The differences between the measured and predicted strain magnitudes were expressed as absolute percentages of the measured values and ranged between 3 and 18%. The limits of agreement between the predicted and measured strain values, as suggested by Bland and Altman (Lancet 1986; 1:307-310), were small enough for the predictions from the FEM to be considered clinically acceptable. The good agreement between the predicted and measured strain values indicates the accuracy of the present FEM. Finite element analysis is a powerful technique that provides a better insight into understanding the complex phenomena of mandibular functional deformation. PMID- 17078049 TI - Identification of a risk haplotype of the alpha-synuclein gene in Japanese with sporadic Parkinson's disease. AB - alpha-Synuclein is one of the main components of Lewy bodies, a pathological marker of Parkinson's disease (PD). Certain missense mutations of the alpha synuclein gene cause familial PD, but the role of the gene in sporadic PD is still controversial. We scrutinized polymorphisms of the alpha-synuclein gene in a Japanese population and investigated their associations with sporadic cases of PD. The 5' flanking region to intron 2 of the alpha-synuclein gene (3.8 kb) and two polymorphisms in intron 4 previously reported in Caucasian sporadic cases of PD were analyzed in 185 sporadic PD and 191 controls. Five novel single nucleotide polymorphisms (SNPs), 16 reported SNPs, and one reported polynucleotide polymorphism (PNP) were found. Most of the polymorphisms examined were in linkage disequilibrium. Significant associations with PD were found in 15 of 21 SNPs, especially in intron 1 (IVS1+155 TmAn PNP and the IVS1+719 C>T SNP, P < 0.0001). Haplotype analysis showed that T10A7-A-A and T11A6-G-G haplotypes at three loci (IVS1+155 - IVS1+273 - IVS1+608) were strongly negative and positive risk factors of sporadic PD, respectively (odds ratios were 0.23 [95% confidence interval, 0.16-0.32] and 1.51 [95% confidence interval, 1.29-1.75]). In conclusion, our findings indicate that genetic variations of the alpha-synuclein gene affect the development of sporadic PD. PMID- 17078050 TI - Hemiballism caused by a small cavernoma in the subthalamic nucleus. PMID- 17078051 TI - Report from a U.S. conference on essential tremor. AB - Seventy researchers met in Washington, DC, on 20-21 October 2005 to identify and discuss the most pressing research issues in essential tremor (ET). The conference attendees concluded that the following six objectives are of immediate and overriding importance: (1) a collaborative network of research centers; (2) an international committee for developing a standard protocol for the diagnosis and quantification of ET; (3) the identification of one or more genes for ET; (4) a centralized repository of DNA and, ideally, immortalized cell lines from well characterized ET families and healthy controls; (5) a reliable and efficient repository of optimally prepared and categorized brain samples for hypothesis driven neuropathological examinations in well-characterized ET patients; and (6) animal models of ET for screening promising drugs. The conference attendees hope that this statement from the United States will engender international collaboration in finding a cure for ET. PMID- 17078052 TI - Radiation-induced xerostomia in patients with head and neck cancer: a literature review. AB - A dry mouth or xerostomia is one of the most common complications during and after radiotherapy for head and neck cancer, because irreparable damage is caused to the salivary glands, which are included in the radiation fields. Xerostomia not only significantly impairs the quality of life of potentially cured cancer patients, it may also lead to severe and long-term oral disorders. Because management of xerostomia is rarely effective, prevention is paramount. Several strategies have been developed to avoid radiation-induced salivary dysfunction without compromising definitive oncologic treatment. These include salivary gland sparing radiation techniques, such as 3-dimensional conformal or intensity modulated radiotherapy, concomitant cytoprotectants, and surgical salivary gland transfer. However, these preventive approaches are not applicable to all patients, and comprehensive scientific research that incorporates new biological insights is warranted to optimize the therapeutic index of radiotherapy for head and neck cancer. PMID- 17078053 TI - Natural history of urothelial inverted papilloma. AB - BACKGROUND: Inverted urothelial papilloma is an uncommon urothelial neoplasm. Although it is traditionally regarded as a benign tumor, conflicting data on multiplicity, recurrence rate, and association with urothelial carcinoma have left uncertainties concerning its biologic behavior. METHODS: The authors analyzed the clinicopathological characteristics of 75 cases of inverted papilloma in the urinary tract without prior or concurrent urothelial carcinoma to determine its biologic behavior and prognosis, and to correlate these findings with surveillance strategies. RESULTS: These patients ranged in age from 26 to 85 years (mean, 60 years). Of the 46 patients for whom tobacco use history was available, 28 gave a history of smoking. Inverted papillomas were located in the urinary bladder (67 cases), prostatic urethra (4 cases), and ureter (4 cases). The majority of vesical tumors arose from the trigone or near the bladder neck. Common presenting complaints included hematuria, dysuria, and irritative voiding symptoms. In 1 case of vesical inverted papilloma, there was a recurrence. All other patients were free of tumor recurrence or progression during a mean follow up of 68 months (range, 2-240 months). CONCLUSIONS: Both the extremely low incidence of tumor recurrence (1%) and strikingly favorable prognosis suggest that inverted urothelial papilloma, when diagnosed according to strictly defined criteria, is a benign urothelial neoplasm not related to urothelial carcinoma. Therefore, complete transurethral resection of inverted papilloma is adequate surgical therapy, and surveillance protocols as rigorous as those employed in the management of urothelial carcinoma seem unnecessary. PMID- 17078054 TI - Nuclear factor-kappaB (NF-kappaB) is frequently expressed in lung cancer and preneoplastic lesions. AB - BACKGROUND: Nuclear factor-kappaB (NF-kappaB), a key transcription factor thought to play a major role in carcinogenesis, regulates many important signaling pathways involved in tumor promotion. Although NF-kappaB can be activated in lung cancer cell lines by tobacco exposure, there have been no studies of the expression of NF-kappaB in lung cancer pathogenesis. METHODS: The immunohistochemical expression of NF-kappaB p65 was investigated in 394 lung cancers (370 nonsmall cell lung carcinomas [NSCLC]; and 24 small cell lung carcinomas [SCLC]) and 269 lung normal epithelium and preneoplastic lesions, including hyperplasias, squamous metaplasias, dysplasias, and atypical adenomatous hyperplasias. RESULTS: High levels of nuclear immunohistochemical expression of NF-kappaB p65 were detected in the lung cancers, with significantly higher levels in SCLCs compared with NSCLCs (P<.0001). In adenocarcinomas the NF kappaB p65 expression level was significantly higher in advanced TNM stages (III IV) than in earlier stages (I-II) (P<.0001), and when NF-kappaB p65 is dichotomized using 50% as the cutoff point (high vs low), a higher NF-kappaB p65 expression level was detected in tumors having either K-RAS (P = .02) or EGFR (P = .009) mutations compared with wildtype tumors. A relatively high level of nuclear NF-kappaB p65 expression was detected in normal and mildly abnormal epithelium, and a progression with increasing histology severity was detected in preneoplastic lesions. CONCLUSIONS: NF-kappaB p65 nuclear expression is an early and frequent phenomenon in the pathogenesis of lung cancer. The findings indicate that NF-kappaB activation plays an important role in lung cancer pathogenesis and is a suitable target for the development of new lung cancer therapies and chemoprevention strategies. PMID- 17078055 TI - Timing of adjuvant chemotherapy initiation after surgery for stage III colon cancer. AB - BACKGROUND: An important advance in medical oncology has been the use of adjuvant chemotherapy for lymph node-positive colon cancer. However, to the authors' knowledge, the effect of the interval between surgery and the initiation of chemotherapy on survival has not been investigated. METHODS: The authors analyzed predictors and outcomes of time intervals to treatment after surgery among patients older than 65 years who were diagnosed with stage III colon cancer between 1992 and 1999 using Surveillance, Epidemiology, and End Results (SEER) Medicare data. Linear and logistic regression analyses were used to model predictors of delay, and Cox proportional hazards models were used to analyze the impact of treatment timing on survival. RESULTS: Among 4382 patients with colon cancer, 1122 patients (26%) began adjuvant chemotherapy within 1 month, 2391 patients (55%) began adjuvant chemotherapy in 1 to 2 months, 454 patients (10%) began adjuvant chemotherapy in 2 to 3 months, and 415 patients (9%) began adjuvant chemotherapy >/=3 months after surgery. Intervals of >/=3 months (delay) were associated with older age, increased comorbid conditions, well/moderately differentiated grade, and being unmarried. Colon cancer-specific mortality was associated with a delay in the initiation of chemotherapy (hazards ratio [HR], 1.48; 95% confidence interval [95% CI], 1.15-1.92), advanced age, increased comorbidity, poorly differentiated tumor grade, the presence of >/=4 positive lymph nodes, and undergoing surgery in a nonteaching hospital. All-cause mortality was associated with intervals >2 months between surgery and chemotherapy (2 to 3 months: HR, 1.41; 95% CI, 1.15-1.74; >/=3 months: HR, 1.62; 95% CI, 1.31-1.99) compared with <1 month. CONCLUSIONS: In the older population that was studied, only 9% of patients initiated adjuvant chemotherapy >3 months after the date of curative surgery. However, delay in initiation was associated with both cancer-specific and all-cause mortality. Determining whether these results were because of chemotherapy timing or other associated factors will require further study. PMID- 17078056 TI - Lay patient navigator program implementation for equal access to cancer care and clinical trials: essential steps and initial challenges. AB - BACKGROUND: Disparities in cancer detection, treatment, and outcomes among racial/ethnic minorities and low-income patients are well documented. One way to reduce these disparities is to use patient navigators to address barriers to care. However, little information about optimal characteristics of navigator programs or considerations for those interested in setting up such programs is available. METHODS: The design and implementation of a patient navigator program for underserved cancer patients in an urban, nonacademic community hospital setting is described. The program, which used lay navigators, was conceived as a component of the Urban Latino African American Cancer (ULAAC) Disparities Project in South Los Angeles, a National Cancer Institute (NCI)-sponsored project to improve cancer care and clinical trial access for minority and low-income patients. RESULTS: Careful initial planning, including input from a community advisory committee, was essential to smooth program implementation. Thirty-one volunteers completed navigator training in the program's first year of operation. Of 135 patients offered navigation services, 75 (56%) accepted, and preliminary feedback from patients, navigators, and providers suggests high levels of satisfaction with navigation. Standardized templates used by navigators and staff to record key information are proving helpful for monitoring quality and outcomes (such as effectiveness in addressing specific barriers to care) and continually improving the program. CONCLUSIONS: The ULAAC program represents a viable model for developing lay navigator programs in community hospitals. Preliminary assessments suggest that the program has a positive effect on minority and low income cancer patients' experience with care and reduces barriers to care. Additional time and research are needed, however, to fully assess the impact on care and outcomes. PMID- 17078057 TI - Telephone monitoring of distress in patients aged 65 years or older with advanced stage cancer: a cancer and leukemia group B study. AB - BACKGROUND: Significant barriers to cancer patients receiving mental health treatment for distress have been reported in the literature. The objective of the current study was to determine whether distress in older patients (aged 65 years and older) would be reduced with educational materials (EM) supplemented by monthly telephone monitoring (TM) (TM + EM) compared with the use of EM alone because of more timely referrals to appropriate health professionals. METHODS: One hundred ninety-two older patients with breast, prostate, and colorectal cancers who had advanced disease and currently were receiving treatment were randomized to receive either TM + EM or EM alone. One hundred thirty-one patients were evaluated by telephone interview for psychologic and physical distress and for social support at baseline and at 6 months using the Hospital Anxiety and Depression Scale (HADS), the European Organization for Research and Treatment of Cancer (EORTC)-QLQ-C30 quality-of-life questionnaire, and the Medical Outcomes Study (MOS) Social Support Survey. Patients who in the TM + EM group were called monthly for 6 months to monitor their distress using the HADS and EORTC physical symptom items and the MOS Social Support Survey items, with cutoff levels were established to indicate which patients were in greater distress. Those patients who scored above the cutoff levels were referred to their oncology nurse for referral to the appropriate professional. Patients in the EM group received written materials regarding cancer-related psychosocial issues and available resources. RESULTS: At 6 months, patients in the TM + EM group reported significantly less anxiety (HADS; P < .0001), depression (HADS; P = .0004), and overall distress (HADS; P < .0001) compared with patients in the EM group. CONCLUSIONS: Monthly monitoring of older patients' distress with TM and EM along with referral for appropriate help was found to be an efficient means of reducing patients' anxiety and depression compared with patients who received only EM. PMID- 17078058 TI - Rapidly progressive sporadic dentatorubral pallidoluysian atrophy with intracytoplasmic inclusions and no CAG repeat expansion. AB - A 48-year-old man developed progressive hemidystonia and postural impairment with falls, followed by choreoathetosis, hyporeflexia, ataxia, supranuclear vertical gaze palsy, and dementia, lasting only 3.5 years from symptom onset to death. Family history and genetic testing were unrevealing. Neuropathology showed findings identical to genetic dentatorubral pallidoluysian atrophy (DRPLA), except for the absence of intranuclear inclusions and the presence of intracytoplasmic inclusions in the pons, striatum, thalamus, and subthalamic nucleus. This case expands the clinical and neuropathological spectrum of DRPLA and supports the hypothesis that aggregates may not be intrinsically pathogenic. PMID- 17078059 TI - Successful antiparkinsonian medication withdrawal in patients with Parkinsonism and normal FP-CIT SPECT. AB - Between 4% and 14% of patients diagnosed with Parkinson's disease and entering clinical trials have normal presynaptic dopaminergic imaging. The effects of antiparkinsonian therapy have varied in these studies, and the consequences of stopping treatment are not reported. We present 11 patients who initially fulfilled diagnostic criteria and were treated for Parkinson's disease but in whom emerging diagnostic doubts led to antiparkinsonian therapy withdrawal, which was achieved without deterioration. Such cases represent a nondegenerative form of Parkinsonism, which does not benefit from dopaminergic therapy. Prospective vigilance regarding this category is of importance in clinical practice and clinical trials. PMID- 17078060 TI - Abnormalities in motor cortical plasticity differentiate manifesting and nonmanifesting DYT1 carriers. AB - A mutation in the DYT1 gene causes dominantly inherited childhood-onset primary dystonia, but intriguingly, only 30 to 40% of those who carry the mutation ever develop symptoms. We have used the unique model provided by this group of patients to investigate the hypothesis that abnormalities in brain plasticity underlie the pathophysiology of primary dystonia. We recruited 8 DYT1 gene carriers with dystonia, 6 DYT1 gene carriers without dystonia, 6 patients with sporadic primary dystonia (torticollis), and 10 healthy control subjects. Groups were age-matched. We compared the effect in these groups of subjects of repetitive transcranial magnetic stimulation (rTMS) delivered to the motor cortex, by assessing changes in corticospinal excitability following rTMS. rTMS was given in the form of theta burst stimulation (TBS) using the inhibitory protocol "cTBS" (total of 300 pulses in 50-Hz bursts given every 5Hz). DYT1 gene carriers with dystonia and subjects with torticollis had a significantly prolonged response to rTMS in comparison with healthy subjects. In contrast, DYT1 gene carriers without dystonia had no significant response to rTMS. These data demonstrate an excessive response to an experimental "plasticity probing protocol" in subjects with dystonia, but a lack of response in genetically susceptible individuals who have not developed dystonia. These preliminary data suggest that the propensity to undergo plastic change may affect the development of symptoms in genetically susceptible individuals and that this may be an important mechanism in the pathogenesis of primary dystonia in general. PMID- 17078061 TI - Inverse drug screens: a rapid and inexpensive method for implicating molecular targets. AB - Identification of gene products that function in some specific process of interest is a common goal in developmental biology. Although use of drug compounds to probe biological systems has a very long history in teratology and toxicology, systematic hierarchical drug screening has not been capitalized upon by the developmental biology community. This "chemical genetics" approach can greatly benefit the study of embryonic and regenerative systems, and we have formalized a strategy for using known pharmacological compounds to implicate specific molecular candidates in any chosen biological phenomenon. Taking advantage of a hierarchical structure that can be imposed on drug reagents in a number of fields such as ion transport, neurotransmitter function, metabolism, and cytoskeleton, any assay can be carried out as a binary search algorithm. This inverse drug screen methodology is much more efficient than exhaustive testing of large numbers of drugs, and reveals the identity of a manageable number of specific molecular candidates that can then be validated and targeted using more expensive and specific molecular reagents. Here, we describe the process of this loss-of-function screen and illustrate its use in uncovering novel bioelectrical and serotonergic mechanisms in embryonic patterning. This technique is an inexpensive and rapid complement to existing molecular screening strategies. Moreover, it is applicable to maternal proteins, and model species in which traditional genetic screens are not feasible, significantly extending the opportunities to identify key endogenous players in biological processes. PMID- 17078062 TI - Short-term effects of tetrabenazine on chorea associated with Huntington's disease. AB - We sought to assess the short-term clinical effects of tetrabenazine (TBZ) on choreic movements in Huntington's disease patients. A total of 10 patients on stable doses of TBZ were enrolled in this observational study. Patients took their evening dose of TBZ and presented the next day to the Baylor College of Medicine Movement Disorders Clinic without taking the usual morning dose. They were assessed using the Unified Huntington's Disease Rating Scale (UHDRS) motor assessment and Beck Depression Inventory. The usual morning dose of TBZ was then administered and patients were followed with serial UHDRS motor examinations approximately every 2 hours until choreic movements subsided and then returned. TBZ decreased the UHDRS chorea score on average 42.4% +/- 17.8%. The duration of effect varied from a minimum of 3.2 hours to a maximum of 8.1 hours (mean = 5.4 +/- 1.3). No patient experienced an adverse event related to TBZ or its withdrawal. During short-term follow-up after a single dose, TBZ improves chorea for approximately 5 hours. PMID- 17078063 TI - Mutations in LRRK2 other than G2019S are rare in a north American-based sample of familial Parkinson's disease. AB - A total of 956 individuals with Parkinson's disease (PD) from 430 multiplex PD pedigrees were screened for 12 previously reported, pathogenic LRRK2 mutations: R793M, L1114L, I1371V, R1441C, R1441G, R1441H, Y1699C, M1869T, I2012T, I2020T, G2385R, and IVS31 +3G > A. Previous screening identified the LRRK2 G2019S mutation in 5% of our families. Only 1 of the 12 newly screened mutations, R1441C, was detected in a single family in our patient cohort. These results indicate that, although the G2019S mutation remains the most common mutation identified in familial PD patients, other mutations in LRRK2 are infrequent. PMID- 17078065 TI - Expression of an Nkx3.1-CRE gene using ROSA26 reporter mice. AB - The genetic locus of Nkx3.1, an early murine marker of sclerotome and prostate development, was disrupted by a knock in of CRE recombinase via homologous recombination in embryonic stem cells. Cell fate mapping revealed previously unidentified cell lineages expanded from Nkx3.1-expressing cell populations and recapitulated reported Nkx3.1 expression patterns. In lineage trace experiments of E18.5 Nkx3.1-CRE; R26R embryos novel staining was observed in areas of the lungs, portions of the duodenum, and vertebral elements of the skeleton. beta galactosidase activity measured in Nkx3.1-CRE; R26R and Nkx3.2-CRE; R26R embryos was observed in overlapping regions of the sclerotome but no apparent change in Nkx3.1 expression was seen in the Nkx3.2 mutants by in situ hybridization. PMID- 17078066 TI - Ectopic expression of Tollo/Toll-8 antagonizes Dpp signaling and induces cell sorting in the Drosophila wing. AB - The wing imaginal disc of Drosophila consists of the primordia for the adult wing and the body wall. The zinc-finger transcription factor Teashirt (Tsh) is expressed in the region proximal to the wing primordium and regulates the formation of the wing-body wall boundary. Here, we report that Tollo/Toll-8, a member of Toll family transmembrane proteins, is also expressed proximal to the wing domain. Ectopic expression of Decapentaplegic (Dpp), a morphogen for wing development, represses tollo expression in the proximal domain. Likewise, misexpression of Tollo in the presumptive wing strongly antagonizes the effects of Dpp signaling. The extracellular domain of Tollo containing the Leucine-Rich Repeats (LRR) is required for the inhibition of Dpp signaling in the wing. Furthermore, clones of cells with Tollo overexpression are sorted out from the surrounding wild-type cells, resulting in the formation of epithelial folds around the clone boundaries. Tsh is ectopically induced at the border of Tollo expressing clones. Despite the strong effects of Tollo overexpression on Dpp signaling and cell sorting, loss-of-function tollo mutants are viable with normal external morphology. Our data suggest that Tollo function might be redundant but is sufficient to antagonize Dpp signaling and induce sorting of Tollo expressing cells from the wing cells to develop proximal cell fate. PMID- 17078067 TI - Computerized posturography balance assessment of patients with bilateral ventralis intermedius nuclei deep brain stimulation. AB - Bilateral ventralis intermedius nuclei (Vim) deep brain stimulation (DBS) improves tremor in patients with both essential tremor (ET) and Parkinson's disease (PD). In each condition, patients have individually noted both subjective improvement and worsening in balance. Computerized posturography (CP) is able to quantify some aspects of balance. Twenty-one patients (8 with PD and 13 with ET) with bilateral Vim DBS were recruited to undergo randomized-order identical CP testing (EquiTest system) while their DBS devices were both activated and deactivated. One PD patient could not complete any OFF assessment and is not included. Three PD patients could not tolerate portions of the OFF testing. Overall, sensory organization testing was improved by DBS activation in conditions that involved quiet standing with eyes open with no motion of the support surface, or with sway-referenced support surface motion, but worsened during quiet standing with eyes closed only in ET patients. Falls lessened with activation in ET patients. Motor control testing was not changed. Bilateral Vim DBS activation mostly improved balance, but may modestly worsen other specific features. PMID- 17078068 TI - Hemiparkinsonism and levodopa-induced dyskinesias following focal nigral lesion. PMID- 17078069 TI - Pisa syndrome without neuroleptic exposure in a patient with Parkinson's disease: A case report. PMID- 17078070 TI - Neurological manifestations in Wilson's disease: Report of 119 cases. AB - We describe the neurological manifestations of 119 patients with WD (93 index cases and 26 affected family members) seen between 1963 and 2004. The mean age at symptoms onset was 19.6 years (range, 7-37 years). Medical records were reviewed for the patient's first neurological examination. The most frequent neurological manifestations observed were dysarthria (91%), gait disturbance (75%), risus sardonicus (72%), dystonia (69%), rigidity (66%), tremor (60%), and dysphagia (50%). Less frequent manifestations were chorea (16%) and athetosis (14%). Rare neurological presentations were seizures (4.2%), and pyramidal signs (3%). PMID- 17078071 TI - Sacral nerve stimulation for voiding dysfunction: One institution's 11-year experience. AB - AIM: The purpose of this study was to review our institution's 11-year experience with SNS for the treatment of refractory voiding dysfunction. Dating back to 1993, it covers a span of time which describes the evolution of SNS as it includes PNE trials, non-tined (bone-anchored or fascial-anchored) leads, percutaneous tined leads with two-staged procedures, and even percutaneous pudendal trials. METHODS: A retrospective review was performed on SNS patients who received an implantable pulse generator (IPG) in our practice from 12/1993 to 12/2004. After Institutional Review Board approval, consents for chart review were obtained from 104 patients, representing 44% of this neuromodulatory patient population. RESULTS: Of our population, 87% were female and 13% were male. Average age at implant was 50 years +/- 13.4 years. Duration of symptoms before implantation was 116 months (range 9-600 months). Eighty percent were implanted for a predominant complaint of urinary urgency and frequency (U/F). Overall, 22% had U/F only, 38% had concomitant urge incontinence (UI), and 20% had concomitant mixed incontinence (MI). Twenty percent were treated for non-obstructive urinary retention (UR), with half of these associated with a neurogenic etiology. Additionally, 46.2% had pelvic pain, 58.6% had bowel complaints, and 51% reported sexual dysfunction. In patients with U/F, mean voiding parameters as described by pre-implant voiding diaries revealed the following: 12.4 (+/-5.1) voids per 24 hr; 2.3 (+/-1.8) voids per night; 5.0 (+/-4.7) leaks per 24 hr; and 2.3 (+/-2.6) pads per 24 hr. Statistically significant improvements post-implantation were noted with mean decreases in the following: 4.3 voids per 24 hr; 1.0 void per night; 4.4 leaks per 24 hr; and 2.3 pads per 24 hr (all P < 0.05). In the UR group a statistically significant improvement post-implantation was noted only in voids per night, with a mean decrease of 0.8 (P < 0.05). With a mean follow up of 22 months (range 3-162 months), sustained subjective improvement was >50%, >80%, and >90% in 69%, 50%, and 35% of patients, respectively. By quality of life survey, 60.5% of patients were satisfied and 16.1% were dissatisfied with current urinary symptoms. Only 13% (14 patients) abandoned therapy, making up a significant portion of those dissatisfied with current urinary symptoms. Good overall lead durability was seen (mean 22 months, range 1-121 months), with the first successful lead proving to be the most durable (mean 28 months, range 1.4 120 months). Lead durability decreased progressively with subsequent trials. Overall, 53% of patients experienced at least one reportable event (RE) attributable to either lead or IPG. A total of 126 REs were noted, with 97% mild to-moderate in severity. REs included lack of efficacy, loss of efficacy, infection, hematoma/seroma, migration, pain, undesirable change in sensation, and device malfunction. In this population, 47.1% of leads were tined while 52.9% were non-tined. Tined leads had an overall lower RE rate as compared to non-tined leads: 28% and 73%, respectively. CONCLUSIONS: SNS is an effective method for treating certain types of voiding dysfunction. Although 53% of patients experienced at least one RE, 97% were mild-to-moderate and did not appear to affect the continued use of this therapy. With improved technology, such as percutaneous tined leads, the RE rate is decreasing. Further analyses of subsets of this population are currently underway. PMID- 17078072 TI - Urinary flow disturbance as an early sign of autonomic neuropathy in diabetic children and adolescents. AB - AIMS: Urinary bladder dysfunction is a frequent complication of diabetes mellitus in adults. The aim of this study is to determine the early disturbances of the detrusor contractility in children, and adolescents with type 1 diabetes. MATERIALS AND METHODS: The bladder urine flow parameters were investigated in 37 children with type 1 diabetes, 10/37 with cardiovascular autonomic dysfunction (CAD) voided 482 +/- 128 mils (Group A), 27/37 without CAD voided 258 +/- 52 mils (Group B). From an earlier uroflow screening study 20 healthy children formed group control A, who had voided 488 +/- 94 mils, and another 30 healthy children formed group control B, who had voided 260 +/- 50 mils. The compared groups were matched for voided volume, age, weight, and height. There was no evidence of urinary tract abnormalities in any of the children. RESULTS: Time to maximum flow was longer in both diabetic groups as compared with controls (P < 0.01), and the acceleration (the ratio of maximum flow and the time to maximum flow) of diabetics was significantly lower (P < 0.01). Bladder emptying was complete in each subject. Bladder wall disturbances were not seen by ultrasound. CONCLUSIONS: The decreased acceleration of detrusor muscle contraction may be interpreted as an early sign of autonomic neuropathy in children and adolescent with type 1 diabetes, even in patients without CAD. Clinicians may be able to determine autonomic neuropathy by using uroflowmetry that is easy to perform, sensitive, reproducible, and needs only a child's minimal cooperation. PMID- 17078073 TI - Spectroscopic study of conformational changes accompanying self-assembly of HCV core protein. AB - Electron microscopy and infrared and Raman spectroscopy have been used here to study the morphology, size distribution, secondary and tertiary structures of protein particles assembled from a truncated hepatitis C virus (HCV) core protein covering the first 120 aa. Particles of pure protein, having similar morphology and size distribution of those of nucleocapsids found in sera from HCV-infected patients, have been visualized for the first time. The secondary structure of these protein particles involve beta-sheet enrichment in relation to its protein monomer. Tertiary/quaternary structure has also been studied using the dynamics of H/D exchange. With this aim infrared spectra were measured as a function of H/D exchange time and subsequently analyzed by principal component analysis and two-dimensional correlation spectroscopy. Temporal dynamics of exchange for these protein particles were as follows: arginine residues exchanged first, followed by turn and unordered structures, followed by beta-sheets which may act as linkers of protein monomers. PMID- 17078074 TI - Interaction of a blood coagulation factor on electrically polarized hydroxyapatite surfaces. AB - Although the polarization treatment of hydroxyapatite (HA) remarkably enhances the osteoconductivity, the mechanisms have not yet been completely understood. The interaction of proteins in blood and tissue fluids with biomaterials are reportedly triggers for later cellular responses and played a major role in osteoconductive processes. Considering this, we disclosed the interaction of polarized HA surface with a coagulation factor, fibrin stabilizing factor XIII (FXIII). The HA activated FXIII even in Ca2+ free buffer, based on the SDS-PAGE detections of alpha-polymer and gamma-dimer bands assigned to stabilized fibrin. The Ca2+ ions, possibly released from the HA surfaces, were examined whether they initiate the activation of the FXIII. It was experimentally proved by ICP analysis that the induced large negative charges on the electrically polarized HA significantly increased the released Ca2+ concentration for the short pre incubation time of 3 min. The more Ca2+ ions released from the negatively charged HA (N-HA) surfaces were more effective in the activation of the FXIII, resulting in the rapider disappearance of the gamma-chain bands in fibrin. The slightly lower Ca2+ concentration in the positively charged HA, compared to the nonpolarized HA activated the FXIII at an almost equal rate. The accelerated activation contributed to the stabilization of fibrin scaffold. Therefore, the polarity difference of the induced charges of the polarized HA surface altered the rate of the FXIII activation. The early stage interaction of the HA surfaces with blood proteins was considered to be an essential process of the accelerated new bone formation near implanted N-HA surface. PMID- 17078075 TI - Crystal structure of the catalytic domain of human DUSP5, a dual specificity MAP kinase protein phosphatase. PMID- 17078076 TI - Free standing carbon nanotube composite bio-electrodes. AB - Carbon nanotubes present a new material for the construction of electrodes for electrochemical devices such as batteries, capacitors, and actuators. Such electrodes require high conductivity, strength, and surface area. The latter two requirements are often incompatible. Electrodes composed entirely of carbon nanotubes (bucky paper) have high surface areas but are typically weak, and have insufficient conductivity for practical macroscopic applications. Here we report a technique that uses naturally occurring biopolymers to produce electrodes (free standing films) that exhibit conductivities of 300 S/cm. These composites also have considerable mechanical strength (up to 145 MPa) and sufficient specific capacitance of 19-27 F/g to enable them to be used as freestanding electrodes. One potential application that deserves special attention is that of biocompatible electrodes, where the binder is a biopolymer already used in a range of implants. Preliminary studies reported here show that the new carbon nanotube biopolymer electrodes can foster prolific L929 cell growth. PMID- 17078077 TI - Optimization of UV cross-linking density for durable and nontoxic collagen GAG dermal substitute. AB - Artificial dermal constructs, based upon collagen-glycosaminoglycan matrices (CGMs), provide new options in treating skin defects. However, their clinical effectiveness may be limited by cytotoxicity related to residual aldehydes left over from the manufacturing process. Although both chemical and dehydrothermal (DHT) cross-linking are used to produce CGMs, we hypothesize that optimized nonchemical cross-linking, using ultra-violet (UV) and DHT treatment combinations, may limit cytotoxicity without sacrificing mechanical strength. Porous CGMs were physically cross-linked using a combination of DHT and varying intensities of UV light. These were compared to glutaraldehyde cross-linked controls. Human keratinocytes were seeded in each matrix, and cellular proliferation measured using a microculture tetrazolium dye assay. A scoring system (based on the in vitro contraction rate, stiffness, and cellular growth of a small cylindrical specimen) was developed to assess the best overall physical cross-linking method. More cellular growth was observed in the 90-120 min UV cross-linked group than in the glutaraldehyde-treated group (p < 0.05). Stiffness was maximized after 0-30 min of UV cross-linking. On the basis of our scoring system, DHT combined with 45 min of UV cross-linking produced the best overall matrix in terms of cellular growth and physical durability. UV cross-linked collagen-based biomaterials could be a viable alternative for use in biological applications to eliminate glutaraldehyde-associated cytotoxicity. PMID- 17078078 TI - Continuous wave ultrasound enhances vancomycin release and antimicrobial efficacy of antibiotic-loaded acrylic bone cement in vitro and in vivo. AB - Although antibiotic-loaded bone cement (ALBC) is used as a drug delivery vehicle to decrease infection rates, the varied clinical effect of the antibiotic release remains controversial. The objective of this study is to investigate the enhancement of continuous wave ultrasound (CWU) on vancomycin release and antimicrobial efficacy of ALBC in vitro and in vivo. We measured vancomycin concentrations after a 0.5-h exposure of CWU. The results showed that CWU increased the drug elution by 2.57-27.44% when compared with the controls in vitro. Ultrasonic intensity and vancomycin load both had a significant effect on the cumulative drug elution at 10.5 h, with a significant interaction between each other. We also implanted ALBC specimens into hip joints of sixteen New Zealand White female rabbits after inoculations of Staphylococcus aureus around primary implants for 30 days. Vancomycin concentrations in the hip cavity and urinary elimination of vancomycin were both measured after intermittent exposures of CWU. The results showed that CWU increased local Tmax by 47.6 microg/mL and urinary elimination of vancomycin by 109.56 microg, but failed to prolong local T>MIC. On day 30 after the implantation, assessment in clinical performance, radiology, bacteriology, and histology all showed a tendency of decreased bacterial vitality and relieved inflammation in the infected hip treated by CWU. This study suggested that CWU could effectively enhance vancomycin release and antimicrobial efficacy of ALBC, which may be of clinical significance for treating prosthesis-related infections. PMID- 17078079 TI - The location and characterisation of the O-linked glycans of the human insulin receptor. AB - O-linked glycosylation is a post-translational and post-folding event involving exposed S/T residues at beta-turns or in regions with extended conformation. O linked sites are difficult to predict from sequence analyses compared to N-linked sites. Here we compare the results of chemical analyses of isolated glycopeptides with the prediction using the neural network prediction method NetOGlyc3.1, a procedure that has been reported to correctly predict 76% of O-glycosylated residues in proteins. Using the heavily glycosylated human insulin receptor as the test protein six sites of mucin-type O-glycosylation were found at residues T744, T749, S757, S758, T759, and T763 compared to the three sites (T759 and T763 correctly, T756- incorrectly) predicted by the neural network method. These six sites occur in a 20 residue segment that begins nine residues downstream from the start of the insulin receptor beta-chain. This region which also includes N linked glycosylation sites at N742 and N755, is predicted to lack secondary structure and is followed by residues 765-770, the known linear epitope for the monoclonal antibody 18-44. PMID- 17078080 TI - Protein adsorption and zeta potentials of a biphasic calcium phosphate ceramic under various conditions. AB - An investigation on the relationship between protein adsorption and zeta potentials of a biphasic calcium phosphate (BCP) ceramic was carried out. Zeta potentials of the BCP ceramic particles were measured at various aqueous solutions. Bovine serum album (BSA) and its competitive adsorption with lysozyme (LSZ) on BCP were investigated using conventional protein adsorption and polyacrylamide gel electrophoresis (PAGE) methods. The results showed that zeta potential and the amount of adsorbed BSA were both influenced by pH, ionic strength, Ca2+ and PO4(3-) concentrations in the buffers. The variation tendencies of BSA adsorption were consistent with that of zeta potentials to some extent. The co-adsorption of BSA and LSZ on BCP was confirmed by the PAGE gel pattern. The semi-quantitative analysis for the detected protein bands proved that LSZ had higher affinity for BCP than BSA and would preferentially bind to the surface. Electrostatic interaction played an important role in protein adsorption on the surface of the BCP ceramic particles. PMID- 17078081 TI - Differential tissue-on-tissue lubrication by ophthalmic formulations. AB - Tissue-on-tissue friction testing was used to determine how instillation of hydrophilic polymer-containing formulations between the "blinking" tissues would compare with lubrication by saline, alone, or an oil-emulsion preparation. Best results were obtained for a formulation that contained active demulcents polyethylene glycol (PEG 400) and propylene glycol (PG), as well as a gellable polymer hydroxypropyl guar (HP-Guar) in a borate-buffered solution, in comparison with hydroxypropylcellulose-containing and carboxymethylcellulose-containing formulations. Superior performance of all the formulations was found for lubricating tissue-on-tissue couples, compared with metal-oxide-to-metal oxide interfaces, or metal oxide-to-tissue interfaces. A reciprocating pin-on-disc type friction/wear test device articulated the intimal faces of preserved human umbilical cord vein segments under increasing loads during simulated continuous "eye-blinking" with addition of increasing weights up to 60 g/cm2, simulating maximal eyelid force on the orbital globe. The tissue-on-tissue couples moved from liquid phase lubrication to boundary lubrication. After residual formulations were rinsed away with saline, persistence of low friction at the highest loads was indicative of formulation substantivity. Human umbilical cord vein segments were utilized in saline-wetted tissue-on-tissue couples that showed variable starting coefficients of friction in the range 0.2-0.4, producing moderate tearing and disruption of the interfacial layers above the medial collagen zone. The best-performing formulations instilled to the tissues pre wetted with saline apparently reacted separately with each tissue face to produce a lower final and persistent coefficient of friction of about 0.05. Scanning electron microscopy and light microscopy of these guar-modified tissue specimens showed only a few superficial tissue disruptions, and some interphase swelling consistent with polymer uptake. The frictional values for lubricated couples having non-tissue members were considerably higher than the coefficients of friction measured for the similarly lubricated tissue-on-tissue couples, emphasizing the requirement that appropriate simulations are critical to obtaining clinically predictive data. PMID- 17078082 TI - Effect of photoactivation protocol and radiant exposure on monomer conversion and flexural strength of a resin composite after water and ethanol storage. AB - The use of soft-start photoactivation to reduce shrinkage stress has become widespread in restorative dentistry. However, an increased susceptibility to ethanol degradation of polymers formed by pulse-delay photoactivation was reported. It was hypothesized that reductions in flexural strength after ethanol storage were related not only to low curing rates, but also to the radiant exposure employed. A commercial composite was subjected to different curing protocols (continuous at high irradiance, continuous at low irradiance, and pulse delay) and radiant exposures (6, 12, and 24 J/cm2). After 48 h, differences in degree of conversion were minimal and no differences in strength were detected among specimens stored in water. Ethanol storage caused significant strength reductions in pulse-delay and low irradiance specimens that received 6 J/cm2. The results suggest that when low irradiances or pulse-delay methods are used, a relatively high radiant exposure is necessary to originate a polymer network structure similar to that obtained by continuous high irradiance photoactivation. PMID- 17078083 TI - Self-setting properties of a beta-dicalcium silicate reinforced calcium phosphate cement. AB - Beta-dicalcium silicate was used to reinforce the injectable calcium phosphate cement (iCPC) for the first time in this study. The influence of the content of beta-dicalcium silicate on the mechanical properties, setting time, rheological properties, injectability, phase evolution, microstructure, and biodegradability of iCPC was systematically investigated. The results demonstrated that the addition of 8 wt % beta-dicalcium silicate obviously enhanced the compressive strength of the CPC from 26.5 to 47.5 MPa, and did not significantly influence the biodegradability, setting time, injectability, phase evolution, and microstructure of the CPC. The beta-dicalcium silicate-reinforced iCPC with relatively high mechanical property should have potential prospects for the wider applications in surgery such as orthopedics, oral, and maxillofacial surgery. PMID- 17078084 TI - Potassium sensitivity test (PST) as a measurement of treatment efficacy of painful bladder syndrome/interstitial cystitis: a prospective study with cyclosporine A and pentosan polysulfate sodium. AB - AIMS: Potassium sensitivity test (PST) has been used as an optional tool in diagnosing painful bladder syndrome/interstitial cystitis (PBS/IC). The role of PST in the follow-up of patients with PBS/IC is elusive. We performed PST before and after treatment of PBS/IC with cyclosporine A (CyA) or pentosan polysulfate sodium (PPS), to test whether the result of repeated PST correlates with alleviation of PBS/IC-related symptoms. MATERIALS AND METHODS: Sixty-four patients who participated in a randomized clinical study comparing CyA and PPS in the treatment of PBS/IC were recruited to the present study. Patients underwent 0.4 M PST before and after 6 months of treatment. The primary end point was a change from positive PST to negative among patients who responded to both treatments determined by global response assessment (GRA). RESULTS: Potassium sensitivity test (PST) was more likely to change from positive to negative among patients who responded to their treatment according to GRA (P < 0.001). The PST change follows the clinical course (ICSI score, voiding frequency, VAS score), which was more beneficial in the CyA-treated patients. CONCLUSIONS: Potassium sensitivity test (PST) reflects symptom severity of PBS/IC patients. Change of pre-treatment positive PST to negative correlates well with clinical alleviation of PBS/IC-related symptoms. In patients with persistent symptoms it may be painful and does not offer additional information. Thus, we do not recommend PST to be used as a routine clinical test in monitoring of PBS/IC treatment efficacy. PMID- 17078085 TI - Addressing thrombogenicity in vascular graft construction. AB - Thrombosis is a major cause of poor patency in synthetic vascular grafts for small diameter vessel (< 6 mm) bypass. Arteries have a host of structural mechanisms by which they prevent triggering of platelet activation and the clotting cascade. Many of these are present in vascular endothelial cells. These mechanisms act together with perpetual feedback at different levels, providing a constantly fine-tuned non-thrombogenic environment. The arterial wall anatomy also serves to promote thrombosis as a healing mechanism when it has been severely injured. Surface modification of synthetic graft surfaces to attenuate the coagulation cascade has reduced thrombosis levels and improved patency in vitro and in animal models. Success in this endeavor is critically dependent on the methods used to modify the surface. Platelets adhere to positively charged surfaces due to their own negative charge. They also preferentially attach to hydrophobic surfaces. Therefore synthetic graft development is concerned with hydrophilic materials with negative surface charge. However, fibrinogen has both hydrophilic and hydrophobic binding sites-amphiphilic materials reduce its adhesion and subsequent platelet activation. The self-endothelializing synthetic graft is an attractive proposition as a confluent endothelial layer incorporates many of the anti-thrombogenic properties of arteries. Surface modification to promote this has shown good results in animal models. The difficulties experienced in achieving spontaneous endothelialisation in humans have lead to the investigation of pre-implantation in vitro endothelial cell seeding. These approaches ultimately aim to result in novel synthetic grafts which are anti thrombogenic and hence suitable for coronary and distal infrainguinal bypass. PMID- 17078086 TI - Role of core support material in veneer failure of brittle layer structures. AB - A study is made of veneer failure by cracking in all-ceramic crown-like layer structures. Model trilayers consisting of a 1 mm thick external glass layer (veneer) joined to a 0.5 mm thick inner stiff and hard ceramic support layer (core) by epoxy bonding or by fusion are fabricated for testing. The resulting bilayers are then glued to a thick compliant polycarbonate slab to simulate a dentin base. The specimens are subjected to cyclic contact (occlusal) loading with spherical indenters in an aqueous environment. Video cameras are used to record the fracture evolution in the transparent glass layer in situ during testing. The dominant failure mode is cone cracking in the glass veneer by traditional outer (Hertzian) cone cracks at higher contact loads and by inner (hydraulically pumped) cone cracks at lower loads. Failure is deemed to occur when one of these cracks reaches the veneer/core interface. The advantages and disadvantages of the alumina and zirconia core materials are discussed in terms of mechanical properties-strength and toughness, as well as stiffness. Consideration is also given to the roles of interface strength and residual thermal expansion mismatch stresses in relation to the different joining methods. PMID- 17078087 TI - Mapping activation levels of skeletal muscle in healthy volunteers: an MRI study. AB - PURPOSE: To use muscle functional MRI (mfMRI) to compare activation levels within and among triceps surae (TS) muscles. MATERIALS AND METHODS: Seven healthy males performed five sets of 10 repetitions of a unilateral heel-raise exercise. T2 weighted images were obtained before and immediately after the exercise. Pixels that showed T2 greater than the mean +1 SD of the region of interest (ROI) in pre exercise images and lower than the mean +1 SD of the ROI in post-exercise images were identified. The remaining T2 values in the post-exercise images were assigned to five categories indicated by color: red (highest level of activation), yellow, green, sky blue, and blue (lowest level of activation). The images were then used to construct three-dimensional (3D) images from which the volumes at each level of activation were determined. RESULTS: Within each of the TS muscles the % activated volumes with low and moderate levels of activation were larger than those with a high level of activation (P < 0.05). The % activated volumes with a high level of activation were larger in the medial gastrocnemius than the soleus (Sol; P < 0.05). The Sol had a larger % activated volume with a low level of activation than the lateral gastrocnemius (P < 0.05). Each activation level was nonuniformly distributed along the length within each TS muscle. CONCLUSION: There is substantial variation in the level of activation within and among the TS muscles; however, the activation level is mainly in the moderate to low range in all three muscles. PMID- 17078088 TI - Variability in RF-induced heating of a deep brain stimulation implant across MR systems. AB - PURPOSE: To compare the MRI-related heating per unit of specific absorption rate (SAR) profile of a conductive implant between two 1.5-Tesla/64 MHz MR systems using a transmit/receive (t/r) head coil configuration. MATERIALS AND METHODS: Deep brain stimulation (DBS) leads were configured within a gel-filled phantom of the human head and torso. Temperature variation at each of four contacts of the bilaterally-placed leads was monitored using fluoroptic thermometry. MRI was performed using the t/r head coils of two different-generation 1.5-Tesla MR systems from the same manufacturer. Temperature changes were normalized to SAR values for the head (DeltaT/SAR-H), and the slope of this DeltaT/SAR-H by time relationship was compared between the two scanners. RESULTS: The DeltaT/SAR-H for the implant ranged from 3.5 to 5.5 times higher on one MR system as compared to the other (P < 0.01) depending on the measurement site. CONCLUSION: The findings support previous observations that console-reported SAR does not constitute a reliable index of heating for elongated, conductive implants, such as the DBS hardware system tested. In contrast to our previous findings using a t/r body coil, the data presented here reveal marked differences between two MR systems using t/r head coils (the coil configuration was consistent with the implant manufacturer's imaging guidelines). J. Magn. Reson. Imaging 2006. (c) 2006 Wiley Liss, Inc. PMID- 17078089 TI - Are there differences between women with urge predominant and stress predominant mixed urinary incontinence? AB - OBJECTIVE: We sought to determine if there are differences in clinical and urodynamic parameters between women with urge predominant and those with stress predominant mixed urinary incontinence (MUI). METHODS: Charts of 99 female patients with complaints of MUI were reviewed. Patients were divided into two groups based on the subjective predominance of either stress incontinence (MSUI) or urge incontinence (MUUI). All patients completed a subjective evaluation including an AUA Symptom Index, Urogenital Distress Inventory (UDI-6), and Incontinence Impact Questionnaire (IIQ-7). Objective non-invasive measures included physical exam, 48-hr voiding diary, and a 24-hr pad test. Videourodynamics studies (VUDS), performed in all patients, were reviewed and the presence and characteristics of detrusor overactivity (DO) and stress incontinence were noted. RESULTS: There were no significant differences between groups with respect to symptom scores. MUUI patients had significantly higher pad usage, and lower maximum and average voided volumes than MSUI patients. They were also more likely to have lower urodynamic bladder capacities and demonstrable DO (70% vs. 26%) on VUDS with contractions occurring at lower bladder volumes and with higher amplitude. MSUI patients were more likely to have demonstrable SUI on physical examination (63% vs. 16%) and on VUDS (100% vs. 61%). CONCLUSIONS: There do appear to be differences in clinical and urodynamic parameters between patients with stress predominant and urge predominant MUI. These may help to determine which component of the mixed incontinence is more problematic. PMID- 17078090 TI - Ca2+-dependent conformational changes in the neuronal Ca2+-sensor recoverin probed by the fluorescent dye Alexa647. AB - Recoverin belongs to the superfamily of EF-hand Ca2+-binding proteins and operates as a Ca2+-sensor in vertebrate photoreceptor cells, where it regulates the activity of rhodopsin kinase GRK1 in a Ca2+-dependent manner. Ca2+-dependent conformational changes in recoverin are allosterically controlled by the covalently attached myristoyl group. The amino acid sequence of recoverin harbors a unique cysteine at position 38. The cysteine can be modified by the fluorescent dye Alexa647 using a maleimide-thiol coupling step. Introduction of Alexa647 into recoverin did not disturb the biological function of recoverin, as it can regulate rhodopsin kinase activity like unlabeled recoverin. Performance of the Ca2+-myristoyl switch of labeled recoverin was monitored by Ca2+-dependent association with immobilized lipids using surface plasmon resonance spectroscopy. When the Ca2+-concentration was varied, labeled myristoylated recoverin showed a 37%-change in fluorescence emission and a 34%-change in excitation intensity, emission and excitation maxima shifted by 6 and 18 nm, respectively. In contrast, labeled nonmyristoylated recoverin exhibited only minimal changes. Time-resolved fluorescence measurements showed biexponentiell fluorescence decay, in which the slower time constant of 2 ns was specifically influenced by Ca2+-induced conformational changes. A similar influence on the slower time constant was observed with the recoverin mutant RecE85Q that has a disabled EF-hand 2, but no such influence was detected with the mutant RecE121Q (EF-hand 3 is nonfunctional) that contains the myristoyl group in a clamped position. We conclude from our results that Alexa647 bound to cysteine 38 can monitor the conformational transition in recoverin that is under control of the myristoyl group. PMID- 17078091 TI - Towards in silico lead optimization: scores from ensembles of protein/ligand conformations reliably correlate with biological activity. AB - Accurately ranking protein/ligand interactions and distinguishing subtle differences between homologous compounds in a virtual focused library in silico is essential in a structure-based drug discovery program. In order to establish a predictive model to design novel inhibitors of dihydrofolate reductase (DHFR) from the parasitic protozoa, Cryptosporidium hominis, we docked a series of 30 DHFR inhibitors with measured inhibition constants against the crystal structure of the protein. By including protein flexibility and averaging the energies of the 25 lowest protein/ligand conformers we obtained more accurate total nonbonded energies from which we calculated a predicted biological activity. The calculated and measured biological activities showed reliable correlations of 72.9%. Additionally, visual analysis of the ensemble of protein/ligand conformations revealed alternative ligand binding pockets in the active site. Using the same principles we then created a homology model of DHFR from Toxoplasma gondii and docked 11 inhibitors. A correlation of 50.2% between docking score and activity validates both the method and the model. The correlations presented here are particularly compelling considering the high structural similarity of the ligands and the fact that we have used structures derived from crystallographic data and homology modeling. These docking principles may be useful in any lead optimization study where accurate ranking of similar compounds is desired. PMID- 17078092 TI - Clinical experience with urethral retro-resistance pressure measurement: a prospective pre- and postoperative evaluation in women with stress urinary incontinence. AB - AIMS: To identify possible correlations of urethral retro-resistance pressure (URP) with clinically and urodynamically proven stress urinary incontinence (SUI) and the outcome of anti-incontinence surgery. MATERIALS AND METHODS: URP was measured using the Monitorr system in women with clinically and urodynamically proven SUI without prolapse before and after anti-incontinence surgery (colposuspension n = 8, tension-free vaginal tape n = 6, tension-free transobturator tape n = 34). RESULTS: Forty-eight women (mean age 61.8 +/- 8.9 years) were evaluated preoperatively and on average 10 weeks postoperatively. Mean URP was 75.6 +/- 20.8 cm H(2)O preoperatively versus 75.4 +/- 17.9 cm H(2)O postoperatively (P = 0.898). The type of anti-incontinence surgery performed had no significant effect on postoperative URP. While no association was found between age and URP (P = 0.35), there was a positive correlation between URP and body mass index (BMI; r = 0.49, P = 0.0004). There was no correlation of URP with the preoperative pad test (P = 0.17) and urethral closure pressure at rest (P = 0.51). Finally, URP did not correlate significantly with the preserved length of the continence zone (0-1/3-2/3-3/3) as determined by the urethral stress profile (P = 0.37-0.72) or with the objective cure rate (negative pad test). CONCLUSIONS: Preoperative URP does not correlate with SUI in all women, has no predictive value, and does not correlate with the outcome of anti-incontinence surgery. However, there seems to be an association with biomechanical factors such as obesity, which may open up a new area of application for URP measurement in urogynecologic diagnosis. PMID- 17078093 TI - An all-atom, distance-dependent scoring function for the prediction of protein DNA interactions from structure. AB - We have developed an all-atom statistical potential function for the prediction of protein-DNA interactions from their structures, and show that this method outperforms similar, lower-resolution statistical potentials in a series of decoy discrimination experiments. The all-atom formalism appears to capture details of atomic interactions that are missed by the lower-resolution methods, with the majority of the discriminatory power arising from its description of short-range atomic contacts. We show that, on average, the method is able to identify 90% of near-native docking decoys within the best-scoring 10% of structures in a given decoy set, and it compares favorably with an optimized physical potential function in a test of structure-based identification of DNA binding-sequences. These results demonstrate that all-atom statistical functions specific to protein DNA interactions can achieve great discriminatory power despite the limited size of the structural database. They also suggest that the statistical scores may soon be able to achieve accuracy on par with more complex, physical potential functions. PMID- 17078094 TI - Long-term benefit to pallidal deep brain stimulation in a case of dystonia secondary to pantothenate kinase-associated neurodegeneration. AB - Pantothenate kinase-associated neurodegeneration (PKAN) is a rare autosomal recessive disorder with onset in childhood and rapid progression. There is no causative and insufficient symptomatic drug therapy. Deep brain stimulation (DBS) of the internal pallidum (GPi) has been reported to improve motor function. Most case reports, however, are limited to short observational periods. The impact of DBS on the progression and life expectancy in PKAN is unknown. We present a 5 year outcome and video documentation of bilateral GPi-DBS of an adolescent patient suffering from genetically defined PKAN. PMID- 17078095 TI - Safety characteristics of gadobenate dimeglumine: clinical experience from intra- and interindividual comparison studies with gadopentetate dimeglumine. AB - PURPOSE: To evaluate the safety and tolerability of gadobenate dimeglumine (Gd BOPTA) relative to that of gadopentetate dimeglumine (Gd-DTPA) in patients and volunteers undergoing MRI for various clinical conditions. MATERIALS AND METHODS: A total of 924 subjects were enrolled in 10 clinical trials in which Gd-BOPTA was compared with Gd-DTPA. Of these subjects, 893 were patients with known or suspected disease and 31 were healthy adult volunteers. Of the 893 patients, 174 were pediatric subjects (aged two days to 17 years) referred for MRI of the brain or spine. Safety evaluations included monitoring vital signs, laboratory values, and adverse events (AE). RESULTS: The rate of AE in adults was similar between the two agents (Gd-BOPTA: 51/561, 9.1%; Gd-DTPA: 33/472, 7.0%; P = 0.22). In parallel-group studies in which subjects were randomized to either agent, the rate of AE was 10.9% for Gd-BOPTA and 7.9% for Gd-DTPA (P = 0.21). In the subset of subjects receiving both agents in intraindividual crossover trials, the rate of AE was 8.0% for Gd-BOPTA and 8.5% for Gd-DTPA (P = 0.84). Results of other safety assessments (laboratory tests, vital signs) were similar for the two agents. CONCLUSION: The safety profile of Gd-BOPTA is similar to Gd-DTPA in patients and volunteers. Both compounds are equally well-tolerated in patients with various disease states undergoing MRI. PMID- 17078096 TI - A comparison of the clinical utility of p16(INK4a) immunolocalization with the presence of human papillomavirus by hybrid capture 2 for the detection of cervical dysplasia/neoplasia. AB - BACKGROUND: Evidence suggests that overexpression of p16(INK4a) protein indicates infection and genomic integration of high-risk human papillomavirus (HR HPV) and predicts progression to cervical high-grade squamous intraepithelial lesions (HSILs) and carcinoma. The authors compared the ability of p16(INK4a) and HR HPV detection by Hybrid Capture 2 (HC2) to detect the presence of significant cervical disease. METHODS.: Four hundred ThinPrep specimens (100 each in 4 categories: 100 specimens that were negative for intraepithelial lesions, 100 specimens of atypical squamous cells of undetermined significance [ASC-US], 100 specimens of low-grade squamous intraepithelial lesions [LSILs], and 100 specimens of HSILs) were analyzed. p16(INK4a) protein was immunolocalized using a specific monoclonal antibody, and the detection of HR HPV in all 400 specimens was determined using HC2. RESULTS: p16(INK4a) was found to be positive in 78% of HSIL specimens, 42% of LSIL specimens, and 36% of ASC-US specimens; whereas HC2 was positive in 92% of HSIL specimens, 81% of LSIL specimens, and 45% of ASC-US specimens. In the HSIL category, the sensitivity, which was calculated using Grade 2 or greater cervical intraepithelial neoplasia as the endpoint, was 78% (50 of 66 specimens) for p16(INK4a) and 91% (60 of 66 specimens) for HC2. For LSIL, the sensitivity was 75% (3 of 4 specimens) for p16(INK4a) and 100% (4 of 4 specimens) for HC2. In the ASC-US category, the sensitivity was 89% (8 of 9 specimens) for p16(INK4a) and 100% (9 of 9 specimens) for HC2. Overall, the sensitivity for HSIL was 92% for HC2 and 78% for p16(INK4a). The specificity for HC2 was 8.3% for HSIL, 16.9% for LSIL, and 48.7% for ASC-US; whereas the specificity for p16(INK4a) was 25% in HSIL, 59.1% in LSIL, and 68.4% in ASC-US. The overall specificity was 25% for HC2 and 56% for p16(INK4a). CONCLUSIONS: Although both p16(INK4a) and HC2 may aid in the clinical management of patients with clinically significant lesions, HC2 was found to have greater sensitivity, and p16(INK4a) greater specificity. The labeling of normal cells and bacteria may preclude the use of p16(INK4a) in automated screening or nonmorphologic assays. PMID- 17078097 TI - Deletion of the chromatin remodeling gene SPT10 sensitizes yeast cells to a subclass of DNA-damaging agents. AB - The Saccharomyces cerevisiae SPT10 protein possesses a DNA-binding domain that is fused to a putative histone acetyltransferase domain. It binds specifically to upstream-activating sequence elements in the core histone promoters and plays a direct role in histone gene regulation. SPT10 is also required for cell-cycle specific K56 acetylation at histone genes, allowing the recruitment of the nucleosome remodeling factor Snf5 and subsequent regulation of gene transcription. We reisolated the SPT10 gene in a functional genome-wide screen designed to identify haploid yeast mutants that are hypersensitive to the antitumor drug bleomycin, which acts by damaging DNA. In addition to bleomycin, we show that spt10Delta mutants are also hypersensitive to a limited set of genotoxic agents that create DNA strand breaks, but not to 254-nm ultraviolet light or 4-nitroquinoline-1-oxide, which generate helix distortion. The hypersensitivities of the spt10Delta mutant to the genotoxic agents are rescued by a single copy plasmid carrying the SPT10 gene. We further showed that spt10Delta mutants displayed a modest twofold increase spontaneous mutant frequency, as compared to the parent. Following exposure to bleomycin, these mutants accumulate unrepaired lesions, e.g., DNA strand breaks with blocked 3' ends in the chromosomal DNA. This defect is not due to the altered expression level or the enzymatic activities of a key DNA repair enzyme, APN1, which is known to repair DNA strand breaks with blocked ends. We propose that SPT10 mediates repair of a subset of DNA lesions by acetylating histones to promote recruitment of DNA repair enzymes. PMID- 17078098 TI - Distribution of paraoxonase-1 gene polymorphisms and enzyme activity in a Peruvian population. AB - Paraoxonase-1 (PON1) is a serum esterase associated with high density lipoproteins and capable of detoxifying toxic metabolites of organophosphorus (OP) compounds. Two major polymorphisms have been described in the coding region of the PON1 gene at positions 192 and 55 and at least five in the 5'-regulatory region, the most important at position -108. Depending on the substrate, PON1 192 Q/R polymorphism can affect PON1 enzymatic activity. In the present study, we have determined the distribution of the PON1 192 Q/R and -108 C/T polymorphisms in a Peruvian population and compared the distribution of these polymorphisms with those of other world populations. PON1 phenotype and enzyme activity also were measured as they can influence the population resistance to the toxicity of OP compounds. The genotype distribution at position 192 was: QQ = 0.236, QR = 0.607, and RR = 0.157; and distribution at position -108 was: CC = 0.315, CT = 0.596, and TT = 0.089. The frequencies of the high activity R and C alleles were 0.461 and 0.613, respectively. The frequency of the PON1 192 Q allele was significantly lower than that of American, Caucasian-American, European Brazilian, and Costa Rican samples. Outside the American continent, the frequency of this allele was lower than for all European countries, Thais, and Indians, but higher than for Chinese or Japanese. Regarding the toxicological importance of these polymorphisms, it was inferred that PON1 phenotyping (assessment of the R alloform) and genotyping (determination of the PON1 -108TT genotype) could be helpful as individual markers of susceptibility. PON1 phenotyping may be useful in further epidemiological studies involving agriculture workers occupationally exposed to OP compounds in developing countries. PMID- 17078099 TI - Genotoxicity in rats treated with the antidiabetic agent, rosiglitazone. AB - Rosiglitazone (RSG), a member of the thiazolidinedione class of antidiabetic agents, improves glycemic control by increasing insulin sensitivity. The therapeutic mode of action of RSG involves its activity as a highly selective and potent agonist for peroxisome proliferator-activated receptor-gamma. Although other drugs in this class have displayed unacceptable hepatotoxicity, RSG was approved for human use. The package insert indicates that RSG has minimal genotoxicity, but information on the genotoxicity of RSG is not available in the published literature. In this study, we used the single cell gel electrophoresis (SCGE)/Comet assay to investigate the DNA damage in peripheral blood and liver cells of rats treated with RSG. Sixteen male Sprague-Dawley rats were randomly distributed into four groups, and dosed daily by oral gavage with 0.0, 0.5, 1.0, and 2.0 mg/kg/day RSG. The rats dosed with 2.0 mg/kg/day RSG received an approximately 10-times the area under the curve concentration of the maximum recommended human daily dose. After 14 days of treatment, the rats were euthanized, and peripheral blood and liver were collected and processed for the Comet assay. A dose-dependent increase in DNA damage (as assessed by % tail DNA and Olive Tail Moment) was observed in the hepatocytes of RSG-treated groups, with significant increases detected between rats treated with all the doses of RSG and the control, and between rats treated with different RSG doses (P < 0.05 P < 0.0001). In contrast, DNA damage was detected in peripheral blood lymphocytes only in rats treated with the higher RSG doses (1.0 and 2 mg/kg/day). Taken together, the data indicate that RSG is able to induce primary DNA damage in rats, with greater damage being detected in liver cells than lymphocytes. PMID- 17078100 TI - Genotoxicity of occupational exposure to wood dust: Micronucleus frequency and nuclear changes in exfoliated buccal mucosa cells. AB - Occupational exposure to wood dust is associated with the occurrence of nasal cancer. In this study, we investigated micronuclei and nuclear changes (NCs: binucleates, karyorrhexis, karyolysis, and the "broken egg" effect) in exfoliated buccal cells of 20 workers exposed to wood dust and 20 age- and sex-matched controls. Micronucleus frequency and the frequency of each of the NCs were significantly higher for wood workers than controls (P < 0.01). Cigarette smoking was associated with increased frequencies of micronuclei and NCs in the buccal mucosa epithelium cells of both the control and exposed groups. Our findings indicate that buccal cells of wood workers display increased levels of genotoxicity and toxicity, and that these biomarker responses may be related to the increased cancer risk among wood workers. PMID- 17078101 TI - Influence of polymorphisms in xenobiotic-metabolizing genes and DNA-repair genes on diepoxybutane-induced SCE frequency. AB - Analysis of the combined effects of polymorphisms in genes encoding xenobiotic metabolizing enzymes (XMEs) and DNA repair proteins may be a key to understanding the role of these genes in the susceptibility of individuals to mutagens. In the present study, we performed an in vitro experiment on lymphocytes from 118 healthy donors that measured the frequency of diepoxybutane (DEB) induced sister chromatid exchanges (SCEs) in relation to genetic polymorphisms in genes coding for XMEs (CYP1A1, CYP2E1, GSTT1, EPHX, and NAT2), as well as DNA repair proteins (XRCC1, XRCC2, XRCC3, XPD, XPA, XPC, XPG, XPF, ERCC1, BRCA1, NBS1, and RAD51). We found that GSTT1(-) and CYP2E1 c1/c2 polymorphisms were associated with higher DEB-induced SCE frequencies, and that NAT2 G(590)A was associated with lower SCE induction by DEB. Analysis of the effect of pairs of genes showed that for a fixed GSTT1 genotype, the SCE level increased with an increasing number of Tyr alleles in EPHX codon 113. We found that among GSTT1(+) individuals the DEB induced SCE level was significantly lower when the EPHX 139 codon was His/Arg rather than His/His. An interaction between polymorphisms in CYP2E1 and at EPHX codon 113 was also observed. The results of our study confirm observations in cancer patients and in people exposed to xenobiotics indicating that sensitivity to mutagens depends upon a combined effect of a variety of "minor impact" genes. Moreover, our results indicate that polymorphisms in genes coding for XMEs have a greater influence on the genotoxic activity of DEB, measured by DEB-induced SCE frequency, than polymorphisms in genes encoding DNA repair proteins. PMID- 17078102 TI - Development of active safety surveillance system for traditional Chinese medicine: an empirical study in treating climacteric women. AB - PURPOSE: Traditional Chinese medicines (TCM) prescribed by doctors are regularly reimbursed by the National Health Insurance (NHI) in Taiwan. The safety of TCM should undergo the same scrutiny that most western medicines do. This study was to monitor adverse events (AEs) associated with a new mixture of TCM, TMN-1, used to treat climacteric symptoms. METHODS: For this multi-center, prospective observational study, we recruited 134 women with climacteric symptoms. During this 12-week study, the subjects made nine visits, took TMN-1 three times a day, and received routine hematologic tests, biochemical tests, and tests for gynecologically relevant hormones at baseline and after 4 and 12 weeks of beginning medication. At every visit, the subjects were asked by questionnaire about any AEs. All AEs were examined through a process of causality assessment (CA) by a research team. RESULTS: In total, we recorded 203 AEs, in order of decreasing incidences--cough, pharyngitis, rhinitis, abdominal pain, abdominal fullness, diarrhea, and pruritus with incidence rates of 2.57, 2.47, 1.88, 1.78, 1.68, 1.58, and 1.58 per 10(3) person-days, respectively. Most AEs were tolerable. Five of the AEs were judged to be "probable" adverse drug reactions (ADRs): Two events of diarrhea and one event each of nausea, abdominal pain, and abdominal fullness. CONCLUSION: This study demonstrates the effectiveness using active safety surveillance to document safety of TCMs. This surveillance system could probably be useful to document the safety of other alternative or complementary medicines. PMID- 17078103 TI - Continuous shipboard sampling system for determination of triple oxygen isotopes and O2/Ar ratio by dual-inlet mass spectrometry. AB - A continuous shipboard sampling system was developed for the determination of the isotopic composition of the triple oxygen isotopes and oxygen to argon (O(2)/Ar) ratios in dissolved air. In this system, dissolved air is separated by a hollow fiber membrane degassing module. This system collects dissolved air quantitatively and rapidly. The sample flow rate through the membrane is critical for the fractionation of the oxygen isotopes and the O(2)/Ar ratio and should be < 2 mL/min. Fractionation of oxygen between the liquid and gas phase of the air saturated water was found to be similar to that of earlier reports. The advantages of this method over existing techniques include rapid collection of samples (30 min/sample), high efficiency in extraction of gases from the liquid phase, and the lack of a sample preparation step (e.g. degassing). PMID- 17078104 TI - Sol-gel zirconia coating capillary microextraction on-line hyphenated with inductively coupled plasma mass spectrometry for the determination of Cr, Cu, Cd and Pb in biological samples. AB - A sol-gel zirconia coating was developed for the preconcentration/separation of trace Cr, Cu, Cd and Pb by capillary microextraction, and the adsorbed analytes were on-line eluted for detection using inductively coupled plasma mass spectrometry (ICP-MS). By immobilizing sol-gel zirconia on the inner surface of a fused-silica capillary, the sol-gel zirconia coating was simply prepared. Its adsorption properties, stability and the factors affecting the adsorption behaviors of Cr, Cu, Cd and Pb were investigated in detail. In the pH range from 7.8 to 10, the zirconia-coated capillary (35 cm x 0.15 mm) is selective towards Cr, Cu, Cd and Pb, and the analyzed ions could be desorbed quantitatively with 0.2 mL of 0.5 mol/L HNO(3) at a rate of 0.2 mL/min. With a consumption of 1.25 mL sample solution, an enrichment factor of 6.25, and detection limits (3sigma) of 9.9 pg/mL Cr, 17.9 pg/mL Cu, 4.5 pg/mL Cd and 3.7 pg/mL Pb were obtained. The precisions for nine replicate measurements of 1 ng/mL Cr, Cu, Cd and Pb were 4.9% Cr, 2.2% Cu, 2.0% Cd and 3.2% Pb (RSD), respectively. The proposed procedure has been applied to the determination of Cr, Cu, Cd and Pb in human urine, which was subjected to microwave-assisted digestion prior to analysis, and the recoveries for these elements were 89.2-101.8%. In order to validate the developed procedure, a NIES No.10-a Rice Flour-Unpolished certified reference material and a BCR No. 184 Bovine Muscle certified reference material were analyzed, and the results are in good agreement with the certified values. PMID- 17078105 TI - Measurement of deamidation of intact proteins by isotopic envelope and mass defect with ion cyclotron resonance Fourier transform mass spectrometry. AB - After synthesis and folding, proteins undergo many post-synthetic modifications, including cleavage, oxidation, glycosylation, methylation, racemization, phosphorylation, and deamidation. Of these modifications, non-enymatic deamidation is the most prevalent. Each asparaginyl and glutaminyl residue in a protein is a miniature molecular clock that deamidates with a genetically determined half-time. These half-times vary from a few hours to more than a century, depending on a primary, secondary, tertiary, and quaternary structure near the amide residue. It has been suggested that these clocks regulate many biological processes. A few such processes have been discovered. These discoveries have been difficult because deamidation is inconvenient to measure. While most post-synthetic changes are easily measured by mass spectrometry, deamidation increases molecular mass by only one nominal Dalton, so the deamidated isotopic envelope overlaps the undeamidated isotopic envelope. While peptide deamidation rate determination through deconvolution of these envelopes has been accomplished for several hundred peptides, deconvolution becomes more difficult as the molecular weight increases. In high-resolution mass spectrometers, this deconvolution is possible for larger molecules and an alternative method based on the 19 mDa mass defect between the deamidated envelope and the isotopic envelope of protein fragments can also be utilized. We herein report a comparison of the envelope deconvolution and the mass defect methods for measurement of deamidation in human eye lens crystallins, with special emphasis on betaB2 crystallin and gammaS crystallin. Measurement of extent of deamidation of betaB2 crystallin in a 7 Tesla ion cyclotron resonance Fourier transform mass spectrometer is found to be accurate to a relative standard deviation in a single measurement of about 4% for each method. The envelope deconvolution method is further illustrated by detection of deamidation in intact gammaS crystallin, a 20 904 Da protein, and discovery of the principal gammaS deamidation site. PMID- 17078106 TI - Generation of end-group information from polyethers by matrix-assisted laser desorption/ionisation collision-induced dissociation mass spectrometry. AB - A series of polyethers, namely poly(ethylene glycol) (PEG), poly(propylene glycol) (PPG), poly(butylene glycol) (PBG) and poly(tetramethylene glycol) (PTMeG), has been characterised by means of matrix-assisted laser desorption/ionisation collision-induced dissociation (MALDI-CID) using a hybrid sector orthogonal-time-of-flight (TOF) instrument. The data indicate that this technique can be used to generate information about the end-group functionality of these polymers, including in some cases information about branching of the alkyl chains of the initiating groups. Proposals are made for the fragmentation pathways for these polymers. PMID- 17078107 TI - The hepatoprotective activity of kinsenoside from Anoectochilus formosanus. AB - Carbon tetrachloride (CCl(4)) causes chronic hepatitis, featuring an increase in hepatic hydroxyproline, spleen weight and serum GPT levels and a decrease in plasma albumin levels. Crude extracts of fresh whole plants of Anoectochilus formosanus showed inhibition of chronic hepatitis induced by CCl(4) in mice. Bioactivity-guided fractionation and spectroscopic analysis revealed that kinsenoside was the most active compound. In an in vitro study, the LD(50) values for H(2)O(2)-induced cytotoxicity in BALB/c normal liver cells were significantly higher after kinsenoside pretreatment than after vehicle alone, further confirming that kinsenoside shows significant antihepatotoxic activity. PMID- 17078108 TI - Pycnogenol increases the probability of the contraction state in chick embryonic cardiomyocytes, indicating inotropic effects. AB - The influence of pycnogenol on the probability of contraction was studied in chick cardiomyocytes. Ventricles from 9-11 day chicken embryos were cultured. After 10-11 days in culture, stable spontaneous contractions were recorded and the contraction kinetics analysed. Isoproterenol and pycnogenol increased the probability of the contraction state. After pretreatment with the beta-receptor antagonist, propranolol reduced the isoproterenol- and pycnogenol-increased probability of contraction state. These data suggested that pycnogenol has inotropic effects via stimulation of beta-receptor mediated activity. PMID- 17078109 TI - Methanol extract of the seaweed Gloiopeltis furcata induces G2/M arrest and inhibits cyclooxygenase-2 activity in human hepatocarcinoma HepG2 cells. AB - It was previously reported that a methanol extract of Gloiopeltis furcata (MEGF), a kind of edible seaweed, inhibited the growth of several human cancer cell lines. In the present study, the effect of MEGF on the growth of human hepatocarcinoma HepG2 cells and its effect on the cyclooxygenases (COXs) expression were investigated. MEGF markedly reduced the viability of HepG2 cells and induced the G2/M arrest of the cell cycle in a concentration dependent manner. These effects were associated with the down-regulation of cyclin A, up regulation of cyclin-dependent kinase (Cdk) inhibitor p21 (WAF1/CIP1) and dephosphorylation of Cdc25C. Furthermore, it was found that MEGF decreased the levels of COX-2 mRNA and protein expression without significant changes in the levels of COX-1, which was correlated with a decrease in prostaglandin E(2) (PGE(2)) synthesis. These findings indicate that MEGF may have a possible therapeutic potential in hepatoma cancer patients. PMID- 17078110 TI - Effects of isoflavones from red clover (Trifolium pratense) on skin changes induced by ovariectomy in rats. AB - Estrogens have a profound influence on skin. The relative hypoestrogenism that accompanies menopause exacerbates the deleterious effects of both intrinsic and environmental aging. Estrogens improve skin in many ways. Among these, they increase collagen content, skin thickness and improve skin moisture. There is evidence that diets with high levels of phytoestrogenic isoflavones are associated with a low incidence of menopausal symptoms and osteoporosis. Plant extracts such as red clover, which contain high levels of isoflavones, have been used to reduce menopausal symptoms and have been shown to reduce bone loss in healthy women. In this study to investigate the effects of red clover isoflavones on skin aging, the histology of the skin, skin thickness and the amount of total collagen determined by a colorimetric method, were studied in ovariectomized rats after treatment for 14 weeks with a red clover extract standardized to contain 11% isoflavones determined by HPLC. In ovariectomized rats the thickness and keratinization of the epidermis were reduced; glands were less in number and vascularity was poor; the distribution and morphology of the collagen bundles and elastic fibers were altered. Whereas the skin of the ovariectomized rats treated with red clover isoflavones (20 and 40 mg of total isoflavones daily for 14 weeks) appeared well organized with a normal epidermis with uniform thickness and regular keratinization; vascularity, collagen and elastic fibers were well developed. The amount of collagen significantly increased in the treated group in comparison with the control group. These findings suggest that red clover isoflavones are effective in reducing skin aging induced by estrogen deprivation. PMID- 17078111 TI - Anethole, a potential antimicrobial synergist, converts a fungistatic dodecanol to a fungicidal agent. AB - Anethole shows synergistic effects on the antifungal activities of phytochemicals including polygodial and (2E)-undecenal against Saccharomyces cerevisiae and Candida albicans. It was found that a fungistatic dodecanol combined with a sublethal amount of anethole showed a fungicidal activity against S. cerevisiae. The MIC of dodecanol quickly reduced cell viability, but the cell viability recovered shortly after and then finally became no longer different from the control, indicating that the effect of dodecanol on this yeast was classified as sublethal damage. On the other hand, anethole completely restricted the recovery of cell viability. Therefore the expression of the synergistic effect was probably due to a blockade of the recovery process from dodecanol-induced stress. PMID- 17078112 TI - Effects of spirulina on the number of ovary mast cells in lead-induced toxicity in rats. AB - The present study investigated the protective effect of Spirulina against the lead-induced increase in mast cells in the ovary during the oestrous cycle of rats. In the ovary cortex and medulla of lead-exposed animals, there was a significant increase in the number of mast cells; however, when also treated with Spirulina, a decrease was observed. The number of mast cells when Spirulina (300 mg/kg) was used alone was not significantly different from that of the control group. These results indicate that Spirulina decreases the number of mast cells induced by lead in the cortex and medulla of rat ovary. PMID- 17078113 TI - Evaluation of the antithyroid, antioxidative and antihyperglycemic activity of scopoletin from Aegle marmelos leaves in hyperthyroid rats. AB - Scopoletin (7-hydroxy-6-methoxy coumarin) was isolated from the leaves of Aegle marmelos and evaluated for its potential to regulate hyperthyroidism, lipid peroxidation and hyperglycemia in levo-thyroxine-induced hyperthyroid rats. Scopoletin (1.00 mg/kg, p.o.) administered daily for 7 days to levo-thyroxine treated animals decreased the levels of serum thyroid hormones and glucose as well as hepatic glucose-6-phosphatase activity, demonstrating its potential to regulate hyperthyroidism and hyperglycemia. Scopoletin also inhibited hepatic lipid peroxidation and increased the activity of antioxidants, superoxide dismutase and catalase. Compared with the standard antithyroid drug, propylthiouracil, scopoletin exhibited a superior therapeutic activity, since unlike propylthiouracil, it also inhibited hepatic lipid peroxidation. These findings indicate that scopoletin has the potential to inhibit thyroid function and hyperglycemia without hepatotoxicity. PMID- 17078114 TI - Mechanisms of endothelin 1-stimulated proliferation in colorectal cancer cell lines. AB - BACKGROUND: The peptide endothelin (ET) 1 promotes proliferation in a number of epithelial cancers. The aim of this study was to identify the mechanism of ET-1 stimulated proliferation in colorectal cancer cells in vitro. METHODS: The effects of ET-1 on colorectal cancer cell lines HT29, LIM1215 and SW620 were studied. Cells were cultured with ET-1 plus antagonists/inhibitors to ET(A) or ET(B) receptors, G protein subtypes, phosphoinositide 3-kinase (PI3K) or protein kinase C (PKC). DNA replication and apoptosis were investigated by 5-bromo-2' deoxyuridine incorporation and Annexin V staining. Transactivation of the epidermal growth factor (EGF) receptor was investigated by blockade of the receptor in the presence of ET-1, measurement of levels of phosphorylated EGF receptor in the presence of ET-1, and comparing the effects of ET-1 and EGF on cell proliferation. RESULTS: ET-1 significantly stimulated growth of all cell lines via ET(A) receptors. ET-1 stimulated DNA replication, not apoptosis. ET-1 stimulated growth was inhibited by antagonism of pertussis toxin-sensitive G proteins, PI3K and PKC. Inhibition of the EGF receptor reduced the effect of ET 1. ET-1 increased levels of phosphorylated EGF receptor via the ET(A) receptor. CONCLUSION: ET-1 increased DNA replication in colorectal cancer cells via the ET(A) receptor. This mitogenic action was mediated via pertussis toxin-sensitive G proteins, PI3K, PKC and transactivation of the EGF receptor. PMID- 17078115 TI - Geographic origin of publications in surgical journals. AB - BACKGROUND: Publications in peer-reviewed journals are the main determinants of research rating and funding. The present study assesses worldwide scientific contributions in the field of surgical research. METHODS: Fifteen major surgical journals were selected for a bibliometric search in Medline/PubMed over a 6-year period (2000-2005). All articles with abstracts were totalled according to country of corresponding author. Publications (total and corrected for population size) and journal impact factor were assessed according to country. RESULTS: A total of 18,717 articles were identified. Fifteen countries generated 88.8 per cent of these: the USA produced 42.1 per cent, Japan 9.1 per cent and the UK 7.6 per cent. When corrected for population size, the Netherlands, Sweden and Switzerland topped the ranking; the USA was sixth. Ireland and Switzerland scored the highest mean impact factor. CONCLUSION: The USA is the most productive country in terms of absolute number of surgical publications in the selected journals. However, when population size is taken into consideration, certain smaller European countries were more prolific. PMID- 17078116 TI - Randomized clinical trial of the effect of postoperative intravenous fluid restriction on recovery after elective colorectal surgery. AB - BACKGROUND: Use of intravenous fluids is an important part of perioperative management. The aim of this study was to compare outcome following administration of restricted or standard postoperative intravenous fluids and sodium in patients undergoing elective colorectal surgery. METHODS: Eighty patients were randomized to restricted fluids (less than 2 litres water and 77 mmol sodium for 24 h after surgery) or a standard postoperative fluid regimen (3 litres water and 154 mmol sodium per day for as long as necessary). The primary endpoint was hospital stay. RESULTS: The median (i.q.r.) total intravenous fluid intake in the restricted group was 4.50 (4.00-5.62) litres compared with 8.75 (8.00-9.80) litres in the standard group (P < 0.001). Intravenous sodium intake was also significantly less in the restricted group (229 (131-332) versus 560 (477-667) mmol; P < 0.001). There was no difference in median time to first flatus (2.9 versus 2.9 days; hazard ratio (HR) 0.85 (95 per cent confidence interval (c.i.) 0.54 to 1.32); P = 0.466) or first bowel motion (4.7 versus 4.9 days; HR 1.06 (95 per cent c.i. 0.68 to 1.65); P = 0.802) between the restricted and standard groups, or in median hospital stay (7.2 versus 7.2 days; HR 1.03 (95 per cent c.i. 0.66 to 1.61); P = 0.902). CONCLUSION: Restriction of postoperative intravenous fluid and sodium does not reduce hospital stay following elective colorectal surgery. PMID- 17078117 TI - A jumping off point. PMID- 17078118 TI - Stand up and be quoted. PMID- 17078119 TI - Snapshots in surgery. Pulsatile scrotum. PMID- 17078121 TI - Russia cracks down on counterfeit drugs. PMID- 17078120 TI - [Prescriptions for psychotropic drugs: results and comments on the 2005 prescription drug report]. PMID- 17078122 TI - India's government aims to improve rural health. PMID- 17078123 TI - Never too old for eating disorders or body dissatisfaction: a community study of elderly women. AB - OBJECTIVE: The aim of the study is to examine eating behavior and body attitude in elderly women. METHOD: A randomly selected nonclinical sample of 1,000 women, aged 60-70 years, was contacted for our questionnaire survey covering current eating behavior, weight history, weight control, body attitude, and disordered eating (DSM-IV). RESULTS: The 475 (48%) women included in our analyses had a mean BMI of 25.1 but desired a mean BMI of 23.3. More than 80% controlled their weight and over 60% stated body dissatisfaction. Eighteen women (3.8%; 95% confidence interval: 2.3-5.9%) met criteria for eating disorders (ED; N = 1 anorexia nervosa, N = 2 bulimia nervosa, and N = 15 EDNOS) and 21 (4.4%) reported single symptoms of an ED. CONCLUSION: Although EDs and body dissatisfaction are typical for young women, they do occur in female elderly and therefore should be included in the differential diagnosis of elderly presenting with weight loss, weight phobia, and/or vomiting. PMID- 17078124 TI - Recruitment for an adolescent bulimia nervosa treatment study. AB - OBJECTIVE: This objective of this study was to investigate the recruitment rate for a clinical treatment trial for adolescents with bulimia nervosa. METHOD: Recruitment rates for a 5-year randomized controlled trial for adolescent bulimia nervosa were analyzed. RESULTS: Although the rate of randomization fluctuated over the course of the study, the overall rate of participants recruited was fairly consistent. The number of participants who were assessed for the study but who did not qualify, and the number of participants who qualified for the study but who chose not to join, were also consistent. CONCLUSION: These findings suggest that it is feasible to recruit adolescents for a bulimia nervosa treatment trial. A better understanding of the reasons behind a participant's decision to stay in a treatment study versus discontinue a treatment study, as well as differences between adult and adolescent patients, could aid in the design of future treatment studies. PMID- 17078125 TI - [Proceedings of the National Conference: The training and information on building safety. Bergamo, Italy, 22 November 2004]. PMID- 17078126 TI - [Proceedings of the National Conference: Research and demonstration of the scientific basis of proof of effectiveness in occupational medicine. Bergamo, Italy, 16 December 2005]. PMID- 17078127 TI - Viewpoint: Gender differences in heart disease. PMID- 17078128 TI - History of cardiology: Henri Louis Roger, MD. PMID- 17078129 TI - Selected papers from the 7th International Conference on Mercury as a Global Pollutant, Ljubljana, SLovenia, June 27-July 2, 2004. PMID- 17078130 TI - Notice of redundant publication. PMID- 17078131 TI - Key factors to consider in choosing an appropriate platform for implementing a learning portfolio. PMID- 17078132 TI - Negative marking can be justified in marking schemes for healthcare professional examinations. PMID- 17078133 TI - Handheld computers are a neglected teaching resource. PMID- 17078134 TI - A teaching ward experience: role blurring is not the answer. PMID- 17078135 TI - Graphic visual adaptation of House-Brackmann facial nerve grading for peripheral facial palsy: an alternative analysis of reliability. PMID- 17078136 TI - The collections treadmill. PMID- 17078137 TI - Prescribing decisions should consider cost. PMID- 17078138 TI - Diagnosis du jour. PMID- 17078139 TI - Explaining the benefits of vaccinations. PMID- 17078140 TI - Universities push their own agendas. PMID- 17078141 TI - Misconceptions in diabetes article. PMID- 17078142 TI - Photos did not empower the elderly. PMID- 17078143 TI - Caring for each other, 'warts and all'. PMID- 17078144 TI - Responses show nurses' prejudices. PMID- 17078145 TI - Support needed for neglected colleagues. PMID- 17078146 TI - Surveying primary health care nurses. PMID- 17078147 TI - [Medication for ADHD and the risk of cardiovascular mortality]. PMID- 17078148 TI - [Xenotransplantation: tour de force or dirty trick?]. PMID- 17078149 TI - In re Michael WW. PMID- 17078150 TI - In re Application of Gribetz. PMID- 17078151 TI - Dwight B. v. Board of Education. PMID- 17078152 TI - Doe v. Roe. PMID- 17078153 TI - Grace Plaza of Great Neck v. Elbaum. PMID- 17078154 TI - [For and against: is quality of life a meaningful indicator of outcome? Against]. PMID- 17078155 TI - Does long-term losartan- vs atenolol-based antihypertensive treatment influence collagen markers differently in hypertensive patients? A LIFE substudy. AB - BACKGROUND: The aim of this study was to investigate the effects of losartan- vs atenolol-based antihypertensive treatment on circulating collagen markers beyond the initial blood pressure (BP) reduction. METHODS: In 204 patients with hypertension and left ventricular (LV) hypertrophy we measured serum concentration of carboxy-terminal telopeptide of type I procollagen (ICTP), carboxy-terminal propeptide of type I procollagen (PICP), amino-terminal propeptide of type III procollagen (PIIINP), amino-terminal propeptide of type I procollagen (PINP) and LV mass by echocardiography at baseline and annually during 4 years of losartan- or atenolol-based antihypertensive treatment; 185 patients completed the study. RESULTS: Beyond the first year of treatment systolic and diastolic BP, LV mass index (LVMI) as well as collagen markers did not change significantly and were equal in the two treatment groups. Changes in PICP during first year of treatment were related to subsequent changes in LV mass index after 2 and 3 years of treatment (r=0.28 and r=0.29, both p<0.05) in patients randomized to losartan, but not atenolol. CONCLUSION: Long-term losartan vs atenolol-based antihypertensive treatment did not influence collagen markers differently, making a BP-independent effect of losartan on collagen markers unlikely. However, initial reduction in circulating PICP may predict later regression of LV hypertrophy during losartan-based antihypertensive treatment. PMID- 17078156 TI - [Atrial fibrillation in an oncological patient with remission]. PMID- 17078157 TI - Characterization of insomnia in patients with essential hypertension. AB - INTRODUCTION: Insomnia may increase risk of cardiovascular events. There is little data available reporting the prevalence and clinical relevance of insomnia in patients with essential hypertension. Therefore, the aim of the study was to investigate the relationship between insomnia and different clinical and biochemical parameters in essential hypertension patients. METHODS: Four hundred and thirty-two patients (mean age 47+/-13 years; 253 male, 179 female) with essential hypertension were screened for insomnia using the athens Insomnia Scale (AIS). Several variables including age, sex, known duration of hypertension, body mass index, creatinine, left ventricular mass index, coexisting disorders, smoking status and alcohol use were analysed. Twenty-four-hour ambulatory blood pressure measurements (ABPM) were performed. RESULTS: Among patients included in the study, 207 subjects (mean age: 49+/-13 years; 47.9%) had an AIS score of 15 or higher and were identified as insomniacs. Insomnia was more frequent in women than in men (60.9% vs 38.7%, p<0.001) and was reported more frequently in patients with coronary artery disease. Subjects with insomnia were older and had longer duration of hypertension. There were no differences between insomniacs and non-insomniacs in ABPM parameters. A relationship was found between the number of antihypertensive drugs and insomnia frequency. There were correlation between AIS score and age (r=0.21; p<0.001) and duration of hypertension (r=0.22; p<0.001). In the sub-group of untreated essential hypertension patients, there were negative correlations between AIS score and night fall in systolic and diastolic blood pressure. CONCLUSION: Our results showed that insomnia is common in patients with essential hypertension and indicate an association between insomnia and gender, known duration of hypertension and number of hypertensive drugs taken. Untreated essential hypertension insomniacs were characterized by less pronounced nocturnal fall in both systolic and diastolic blood pressure compared with non-insomniacs. PMID- 17078158 TI - [Spontaneous mesenteric hematoma of unknown origin]. PMID- 17078159 TI - [Double cardiac rupture post myocardial acute infarction]. PMID- 17078160 TI - [Crohn's disease and prosthetic hip infection due to an enteric fistula]. PMID- 17078161 TI - [Patient with vertigo, frustrated situation for the doctor and patient]. PMID- 17078162 TI - [Sacroiliitis and Behcet disease]. PMID- 17078163 TI - [Synchronous diagnosis of small-cell lung cancer and Hodgkin lymphoma]. PMID- 17078164 TI - Motherhood in Black and White: race and sex in American liberalism, 1930-1965. [Review of: Feldstein, R. Motherhood in Black and White: race and sex in American liberalism, 1930-1965. Ithaca, N.Y.: Cornell U. Pr., 2000]. PMID- 17078165 TI - The Mixed legacy of Charlotte Perkins Gilman. [Review of: Golden, C. J. and Zangrando, J. S., ed. The Mixed legacy of Charlotte Perkins Gilman. Newark: U. of Delaware Pr., 2000]. PMID- 17078166 TI - Women in labor: mothers, medicine, and occupational health in the United States, 1890-1980. [Review of: Hepler, A.L. Women in labor: mothers, medicine, and occupational health in the United States, 1890-1980. Columbus: Ohio State U. Pr., 2000]. PMID- 17078167 TI - Teaching sex: the shaping of adolescence in the 20th century. [Review of: Moran, J.P. Teaching sex: the shaping of adolescence in the 20th century. Cambridge, Mass.: Harvard U. Pr., 2000]. PMID- 17078168 TI - A tale of two drugs. PMID- 17078169 TI - New bill aims to reform US patent system. PMID- 17078170 TI - HIV prevention trials in danger of grinding to a standstill. PMID- 17078171 TI - William R. Keller. Interview by Sabine Louet. PMID- 17078172 TI - Life after statin patent expiries. PMID- 17078173 TI - Ranibizumab. PMID- 17078174 TI - Inner hygiene: constipation and the pursuit of health in modern society. [Review of: Whorton, J.C. Inner hygiene: constipation and the pursuit of health in modern society. New York: Oxford U. Pr., 2000]. PMID- 17078175 TI - Permanent waves: the making of the American beauty shop. [Review of: Willett, J.A. Permanent waves: the making of the American beauty shop. New York: New York U. Pr., 2000]. PMID- 17078176 TI - Assessment of self-monitoring of blood pressure in the diagnosis of isolated clinic hypertension. AB - BACKGROUND: There are no studies assessing cardiovascular morbidity, morality in patients with isolated clinical hypertension (ICH) with self-blood pressure monitoring (SBPM). OBJECTIVES: To determine the value of SBPM in the diagnosis of ICH. METHODS: Cohort study. New hypertensive and normotensive patients 15-75 years, without cardiovascular events history. VARIABLES: Oriented anamnesis hypertension; blood pressure measurements (BP): clinical BP, SBPM and ambulatory BP monitoring (ABPM); evaluation of target organ damage (TOD); electrocardiogram; retinography and microalbuminuria (MA). RESULTS: One hundred and thirty-five patients, 95 hypertensive (62.1% males; mean age 59.08+/-16.8 years), 40 normotensive (37.5% males; mean are 56.32+/-10.22 years). BP measurements (mmHG) in normotensives vs hypertensives: clinical BP, 125.36/76.74 vs 149.81/87.86 mmHg (p<0.0001) and SPPM, 114.90/69.96 vs 142.06/86.31 (p<0.0001). Twenty-four-hour ABPM: 135.41/81/81.74. Prevalence of TOD in hypertensive: 23.10% left ventricular hypertrophy (LVH), sustained hypertension (SH): clinic BP, 149.88/86.34 vs 152.51/89.55 (p>0.10); SBPM: 147.895/88.95 vs 128.17/79 (p<0.0001) and ABPM, 141.72/88.22 vs 131.66/80 (p=0.053 for systolic). TOD in SH vs ICH: LVH, 24.6% vs 19.2% (p=0.814); exudates or haemorrhages, 7.7% vs 9.8% (p=0.580). The risk of an occurrence of any TOD in ICH patients is lower for 125/80 (OR=2.5). CONCLUSIONS: VAMPAHICA will provide information about value SBPM in the diagnosis of ICH. Advanced retinopathy is relative frequent in ICH patients. If TOD is accepted as a surrogate endpoint, the diagnostic values of ICH will be probably decreased. PMID- 17078177 TI - American health care: government, market processes, and the public interest. [Review of: Feldman, R. D., ed. American health care: government, market processes, and the public interest. New Brunswick, N.J.: Transaction, 2000]. PMID- 17078178 TI - Estimated risk of a first stroke and conditioning factors in Spanish hypertensive women. The RIMH study. AB - Stroke is the leading cause of mortality in women in Spain. RIMHA is a cross sectional multicenter study in hypertensive women aged 55 or more in primary care to estimate the 10-year risk for a first stroke. Clinical history, cardiovascular risk factors and diseases, electrocardiogram, blood samples and blood pressure (BP) were recorded. Stroke and coronary risk were estimated using the appropriate Framingham scales; 12875 patients were included (mean age 68.0+/-8.5 years, 29.1% with diabetes, 19.7% with cardiovascular disease). Electrocardiographic left ventricular hypertrophy (LVH) was present in 19.2% BP was controlled in 42.9% of non-diabetic (BP<140/90 mmHg) and 9.7% of diabetic patients (BP<130/80 mmHg). The 10-year risk (+/- SD) for a first stroke was estimated as 15.8+/-16.3%, and the coronary risk as 12.0+/-6.3. In the multivariate analysis, the most contributing factors for stroke risk estimation were age, systolic BP, LVH and atrial fibrillation. In conclusion, the 10-year estimated stroke risk for Spanish hypertensive women aged 55 years or more was higher than the estimated coronary risk, in accordance with the high rates of morbidity and mortality due to stroke among women in Spain. The most powerful risk factors were older age, poor BP control, LVH and atrial fibrillation. PMID- 17078179 TI - Don't breathe the air: air pollution and U.S. environmental politics, 1945-1970. [Review of: Dewey, S.H. Don't breathe the air: air pollution and U.S. environmental politics, 1945-1970. College Station: Texas A. & M. U. Pr., 2000]. PMID- 17078180 TI - Multiple publication reconsidered. PMID- 17078181 TI - A social contract for the coal fields: the rise and fall of United Mine Workers of America Welfare and Retirement Fund. [Review of: Mulcahy, R.P. A social contract for the coal fields: the rise and fall of United Mine Workers of America Welfare and Retirement Fund. Knoxville: U. of Tennessee Pr., 2000]. PMID- 17078182 TI - A social contract for the coal fields: the rise and fall of United Mine Workers of America Welfare and Retirement Fund. [Review of: Mulcahy, R.P. A social contract for the coal fields: the rise and fall of United Mine Workers of America Welfare and Retirement Fund. Knoxville: U. of Tennessee Pr., 2000]. PMID- 17078183 TI - Predictors for high costs of hospital care in elderly hypertensive patients. AB - OBJECTIVE: To analyse predictors of high cost of care in elderly hypertensive patients, in particular costs related to short-term (<10 days) and long-term (< or = 10 days) institutional care. DESIGN: Health Economy (HE) sub-study in the Swedish Trial in Old Patients with Hypertension-2 (STOP Hypertension-2). SETTING: Outpatient clinics, hospitals and nursing homes in Sweden. SUBJECTS: Elderly (70 84 years) patients (n=6614) from the STOP Hypertension-2 cohort with a systolic or diastolic hypertension, or a combination thereof, were included. METHODS: Costs of institutional care were analysed and categorized as short-term (<10 days or long-term care (> or = 10 days). Costs were related to individual patients and calculations were made during follow-up in STOP Hypertension-2 from inclusion to end of study. Data was available from 99% of all scheduled visits during the median 5.3 years of follow-up in the 6614 elderly hypertensive patients. RESULTS: A multivariate analysis of potential predictors for inpatient short-term or long term care demonstrated that several clinical factors within the groups of target organ damage (TOD), associated clinical conditions (ACC), as well as additional risk factors (RF) predicted for an increased probability of inpatient care in elderly hypertensives. Specifically, predictors for heart failure (OR 1.73, p=0.005), diabetes (OR 1.36, p<0.0005) and older age (OR 1.05, p<0.0001). Predictors at entry for long-term care (> or = 10 days) were; presence of ischaemic heart disease (OR 1.65, p<0.0001), diabetes mellitus (OR 1.32, p=0.012), female gender (OR 0.80, p=0.0003) as well as older age (OR 1.02, p=0.046). High total costs for this cohort of elderly hypertensive patients were recorded in the group subjected to long-term care for cardiovascular as well as non-cardiovascular reasons. Male gender (p=0.004) and stroke (p=0.06) remained predictors for high costs for hospital care while stroke (p<0.0001) and old age (p<0.0001) predicted for high costs for nursing home care. CONCLUSION: In elderly hypertensives in STOP Hypertension-2, presence of cardiac disease, stroke, diabetes and older age at entry increased the probability as well as costs for both short- and long-term care. Level of systolic or diastolic blood pressure did not predict for hospitalization or cost outcome. Our results provide an economic argument for strict risk reduction focus in the management of elderly high-risk hypertensive patients. PMID- 17078184 TI - Shaping biology: the National Science Foundation and American biological research, 1945-1975. [Review of: Appel, T. Shaping biology: the National Science Foundation and American biological research, 1945-1975. Baltimore: Johns Hopkins U. Pr., 2000]. PMID- 17078185 TI - A new and untried course: Woman's Medical College and Medical College of Pennsylvania, 1850-1998. [Review of: Peitzman, S. J. A new and untried course: Woman's Medical College and Medical College of Pennsylvania, 1850-1998. New Brunswick, N.J.: Rutgers U. Pr., 2000]. PMID- 17078186 TI - Beaches, blood, and ballots: a black doctor's civil rights struggle. [Review of: Mason, G. R.; Smith, J. P. Beaches, blood, and ballots: a black doctor's civil rights struggle. Jackson: U. Pr. of Mississippi, 2000]. PMID- 17078187 TI - Control of hypertension in patients with peripheral artery disease. PMID- 17078188 TI - Prevention of type 2 diabetes mellitus with antihypertensive drugs. PMID- 17078189 TI - Treatment of hypertensive urgencies and emergencies. PMID- 17078190 TI - Skull wars: Kennewick Man, archaeology, and the battle for Native American identity. [Review of: Thomas, D. H. Skull wars: Kennewick Man, archaeology, and the battle for Native American identity. New York: Basic, 2000]. PMID- 17078191 TI - Tobacco use by Native North Americans: sacred smoke and silent killer. [Review of: Winter, J.C., ed., Tobacco use by Native North Americans: sacred smoke and silent killer. Norman: U. of Oklahoma Pr., 2000]. PMID- 17078192 TI - Final diagnosis: polymigratory arthritis, gastrointestinal ulcers, cholecystitis, and severe hypoalbuminemia secondary to Henoch-Schonlein purpura. PMID- 17078193 TI - Consensus statement on surgery journal authorship--2006. PMID- 17078194 TI - Colonial intimacies: Indian marriage in early New England. [Review of: Plane, A.M., Colonial intimacies: Indian marriage in early New England. Ithaca, N.Y.: Cornell U. Pr., 2000]. PMID- 17078195 TI - Colonial intimacies: Indian marriage in early New England. [Review of: Plane, A.M., Colonial intimacies: Indian marriage in early New England. Ithaca, N.Y.: Cornell U. Pr., 2000]. PMID- 17078196 TI - They say he was witched. PMID- 17078197 TI - Repatriation reader: who owns American Indian remains? [Review of: Mihesuah, D.A., ed. Repatriation reader: who owns American Indian remains? Lincoln: U. of Nebraska Pr., 2000]. PMID- 17078198 TI - Public vows: a history of marriage and the nation. [Review of: Cott, N.F. Public vows: a history of marriage and the nation. Cambridge, Mass.: Harvard U. Pr., 2000]. PMID- 17078199 TI - Suffering soldiers: Revolutionary War veterans, moral sentiment, and political culture in the early Republic. [Review of: Resch, J. Suffering soldiers: Revolutionary War veterans, moral sentiment, and political culture in the early Republic. Amherst: U. of Massachusetts Pr., 2000]. PMID- 17078200 TI - Immigrant mothers: narratives of race and maternity, 1890-1925. [Review of: Irving, K. Immigrant mothers: narratives of race and maternity, 1890-1925. Urbana: U. of Illinois Pr., 2000]. PMID- 17078201 TI - Cocaine: from medical marvel to modern menace in the United States, 1884-1920. [Review of: Spillaine, J. F. Cocaine: from medical marvel to modern menace in the United States, 1884-1920. Baltimore: Johns Hopkins U. Pr., 2000]. PMID- 17078202 TI - Cocaine: from medical marvel to modern menace in the United States, 1884-1920. [Review of: Spillaine, J. F. Cocaine: from medical marvel to modern menace in the United States, 1884-1920. Baltimore: Johns Hopkins U. Pr., 2000]. PMID- 17078203 TI - [The Twenty-third National Surgical Congress, May 24-27, 2006, Felix-Oradea]. PMID- 17078204 TI - Metaplasia--a transdifferentiation process that facilitates cancer development: the model of gastric intestinal metaplasia. AB - Metaplasia, in general, and intestinal metaplasia, in particular, are transdifferentiation processes triggered by environmental aggressions. The heterogeneity of different morphologic variants of metaplasia and distinction from close morphologic entities--heterotopia and hamartoma--is reviewed. Recent data on the characterization of the molecular and epidemiological data involved in the development and progression of lesions of gastric intestinal metaplasia are discussed. A redefinition of intestinal metaplasia is proposed. PMID- 17078205 TI - Sessile serrated adenoma: a brief history and current status. AB - Sessile serrated adenoma (SSA) is a polyp of the large intestine, which was first described in 1996. It presents as a solitary lesion or in a setting of a polyposis previously coined as "serrated adenomatous polyposis." The importance of correct recognition of this lesion and distinction from other serrated polyps of the colon is in that the newly described "serrated pathway" of colorectal carcinogenesis seems to apply mostly to SSA and not to other serrated polyps of the colon. This review describes the history of morphologic discovery of SSA and reports on current status of this lesion. PMID- 17078206 TI - Methods for detection of subtle mutations in cancer genomes. AB - With the realization that cancer is a genetic disease, detection of mutations in genomic DNA has become an important discipline in many areas of cancer research. Although the publication of the human genome sequence and the immense technological advancements have facilitated the analysis of cancer genomes, detection of mutations in tumor specimens may still be challenging and fraught with technical problems. In this review, we describe current technologies for detection of small DNA mutations, including mutation scanning techniques to search for unknown mutations, and diagnostic techniques to detect known cancer mutations. We outline the principles of the different techniques and discuss their advantages and limitations. We also discuss critical issues that must be considered before choosing methodology, including sensitivity, specificity, limit of detection, throughput and cost, quantity and quality of template DNA, available equipment, and personnel expertise. PMID- 17078207 TI - Familial breast cancer, underlying genes, and clinical implications: a review. AB - Family history has been a recognized risk factor for breast cancer for many years. We will, in this review, describe the research field in breast cancer susceptibility from the early epidemiological, loss of heterozygosity (LOH), and linkage studies in the 1990s leading to the discovery of BRCA1 and 2, and the subsequent search for other high- and low-risk genes at present. We will also describe the clinical implications of BRCA1 and 2. Finally, we will give a review of the field but also focus on our own research, molecular and clinical. PMID- 17078208 TI - Methyl-group metabolism and the response of colorectal cancer to 5-fluorouracil. AB - The major mechanism of action of 5-fluorouracil (5FU)-based therapies is thought to be inhibition of thymidylate synthase (TS). This enzyme catalyzes synthesis of the thymidine nucleotide precursor using a methyl group provided by a folate cofactor. In addition to TS activity levels, various elements of methyl-group metabolism could also be predictive for the response of colorectal cancer (CRC) to 5FU. These include the activity of enzymes involved in folate metabolism, the concentrations of intracellular folate intermediates, and surrogate markers of aberrant methyl-group metabolism, such as DNA methylation and microsatellite instability. The factors of age, gender, common genetic variants, and diet have been shown to influence both systemic and tumor methyl-group metabolism. This has important implications for the prediction of toxicity and response to 5FU, respectively. Identification of predictive factors within the methyl-group metabolism pathway should assist in targeting 5FU treatment to the most responsive CRC patient groups. This is particularly important for early-stage disease where conclusive demonstration of a survival benefit from 5FU in the overall CRC group has thus far proven difficult. PMID- 17078209 TI - siRNA: a potential tool for future breast cancer therapy? AB - The siRNA technology is a promising research tool for use in functional genomics, and it also shows potential for use in future therapy. Thus far, siRNAs have been used to specifically silence disease-associated alleles in animal models. The technology is still quite new, and the biological mechanisms underlying RNAi and siRNA-mediated knockdown of gene expression are not yet fully understood. The main issues when siRNAs are designed are efficiency and specificity, and it is of great importance to consider possible off-target effects in the siRNA design. One major challenge in siRNA-based therapy is the development of systems for efficient delivery to the target cells. A large number of tools have, over the last few years, been designed for the delivery of DNA and RNA for gene therapy, and extensive efforts are now placed into developing clinical applications of siRNAs in a range of human diseases, including breast cancer. PMID- 17078210 TI - [Some features of cardiac arrythmia treatment in outpatient practice]. PMID- 17078211 TI - [Is triple combination of different neurohormonal modulators recommended for treatment of mild-to-moderate congestive heart failure patients? (Results of Sadko-CHF study)]. AB - AIM: To assess effects of different variants of neurohormonal (NH) modulation with angiotensin converting enzyme (ACE-I) quinapril (Q), angiotensin-receptor blocker (ARB) valsartan (V) and their combination in addition to beta-adrenergic blocker bisoprolol (B) on functional status, quality of life (QOL), parameters of left ventricular (LV) remodeling, main indices of 24-h heart rate variability (HRV) and NH profile in patients with stable mild-to-moderate congestive heart failure (CHF). MATERIAL AND METHODS: Sixty three patients with CHF (NYHA class II III) as a result of ischemic heart disease and dilated cardiomyopathy with LV EF < 40% were randomly assigned to one of the treatment variants on 1:1:1 basis: B+Q (n = 22), B+V (n = 23) and combination of B+Q + V (n = 18). At baseline, all the patients in this study were on background B treatment and according to the study design Q or V were then added to B at randomization. NYHA FC, 6-min walking test (6MT), QOL, 2D-echocardiography, plasma renin activity (PRA), angiotensin II (AT II), aldosterone (Ald), norepinephrine (NE), epinephrine (E), brain natriuretic peptide (BNP) concentrations and 24-hour HRV parameters were investigated at baseline, 3 and 6 months after randomization. RESULTS: During the study NYHA FC improvement was revealed in all three treatment groups with comparative significant changes in 6MT distance by 20.4%, 19.1% and 19.4% in B+Q, B+V and B+Q+V groups, respectively. QOL maximally decreased in B+V combination (from 45 to 21 points). LV volumes significantly decreased and LV ejection fraction (EF) increased in all groups to the end of the study. Triple combination had no additional effect on LV volumes and LVEF changes compared to B+Q and B+V groups. Plasma NE concentrations decreased maximally in B+Q group (from 650 to 430 pg/ml, p = 0.007). The lesser effect was observed in the combination of B+Q+V, with any NE changes in B+ V group. The E concentration increased significantly (from 215 to 295 pg/ml, p = 0.024) in the B+Q+V group at the end of the study. Plasma A-II concentration did not differ from the baseline during the study in B+Q group, but significantly increased in B+V group and maximally in B+Q+V group (from 11.4 to 23.5 pg/ml, p = 0.009). To the end of the study plasma Ald concentrations remain reduced significantly only in B+V group. The level of BNP significantly decreased in all 3 treatment groups. Significant changes in HRV indices, both in time and frequency domain, were revealed in the B+Q group at 3-month follow-up and SDNN increased on month 24 (p = 0.039). These changes became insignificant at the end of the study. The lesser effect was revealed in B+Q+V group, with insignificant trend toward an increase of SDNN to the end of the study. HRV indices did not improve in the B+V group. CONCLUSION: During long-term treatment the triple combination of B+Q+ V has no significant advantages over B+Q and B+V by the functional status, QOL and parameters of LV remodeling in patients with mild-to moderate CHF. The combination of B+Q has more potent effect on 24-hour HRV parameters, sympatho-adrenal activity and renal function compared to B+V and B+Q+V groups in CHF patients in our study. The combination B+Q+V may have a negative effect on NH profile (excessive activation of ATII and E) in CHF patients. The triple combination is not recommended for therapy of stable mild-to moderate CHF patients. PMID- 17078212 TI - [Clinicofunctional characteristics of patients with ischemic heart disease after coronary artery bypass surgery as shown by 5-year follow-up]. AB - AIM: To assess immediate and 5-year follow-up results of coronary artery bypass surgery (CABS) in patients with ischemic heart disease (IHD). MATERIAL AND METHODS: Psychophysiological state (severity of myocardial lesion, psychological status, blood lipids) was evaluated in 58 IHD males aged 44 to 59 (mean age 53.7 +/- 4.2 years). The above psychophysiological parameters were studied before grafting and 3 months, 1 year, 3 years and 5 years after it. RESULTS: In patients with a considerable myocardial reserve, CABS resulted in a rapid and adequate restoration of the left ventricular function, good exercise tolerance, long compensation. Low myocardial reserve secondary to atherosclerotic and postinfarction cardiosclerosis was associated with a low effect of intracardiac and central hemodynamic rearrangement. Patients with marked hypercholesterolemia had a positive effect of CABS but anginal manifestations in them came earlier and were stronger. Psychic condition of the patients depended on their somatic state determined by the course of the underlying disease. CONCLUSION: Myocardial function recovery depends not only on revascularization but also on its initial condition. This should be taken in consideration in long-term monitoring of patients after CABS. PMID- 17078213 TI - [Comparative efficacy of carvedilol and capoten in the treatment of an uncomplicated hypertensive crisis]. AB - AIM: Efficacy and tolerability of carvedilol vs captopril were studied at 24-h monitoring of blood pressure (BP) in hypertensive patients with an uncompletated hypertensive crisis (HC). MATERIAL AND METHODS: The trial included 43 patients (23 males and 20 females aged 58 +/- 4.1 years) with an uncomplicated HC (duration of hypertension 9.4 +/- 1.1 years). Twenty patients of group 1 received oral carvedilol (25 mg), 23 patients of group 2--oral captopril (25 mg). The criterion of efficacy was a 15-25% decrease of BP within 60-120 min. RESULTS: Attenuation of HC symptoms and tolerability was the same in both groups. Forty five minutes after the drug intake systolic pressure lowered by 11.1% (p = 0.039) and 10.9% (p = 0.042) in group 1 and 2, respectively; diastolic pressure--by 14.9% (p = 0.037) and 17.9% (p = 0.018), respectively. Heart rate diminished significantly only in group 1 (by 10.9%, p = 0.043) 30 min after carvedilol intake. A maximal BP fall in group 1 occurred 180 min after the drug intake: systolic by 23.5% (p = 0.0001), diastolic by 26.9% (p < 0.0001). In group 2 the BP fall was 23.3% (p < 0.0001) and 29.1% (p < 0.0001) on min 165 and 150, respectively. Systolic and diastolic pressures lowered faster in group 1. The effect of carvedilol lasted longer--372.6 +/- 19.3 min and 245.1 +/- 13.7 min, respectively, p = 0. 0001. CONCLUSION: A hypotensive effect of carvedilol and captopril in hypertensive patients with an uncomplicated HC was the same. Carvedilol produced a longer and a more stable effect. PMID- 17078214 TI - [Characteristics of action of various drugs blocking atrioventricular conduction (beta-blockers, verapamil, diltiazem) in constant fibrillation tachyarrhythmia. Is monotherapy optimal?]. AB - AIM: To characterize actions of beta-blockers and Ca antagonists (verapamil and diltiazem) on the rate, structure and parameters of ventricular rhythm variability in constant cardiac fibrillation (CCF) and to evaluate validity of monotherapy with these drugs. MATERIAL AND METHODS: Thirty patients with CCF (mean age 64.5 +/- 9.5 years) received beta-blockers (n = 10, atenolol in a dose 50.0 +/- 23.2 mg/day or metoprolol in a dose 45.0 +/- 20.9 mg/day), verapamil (n = 10, 192.0 +/- 83.9 mg/day) and diltiazem (n = 10, 286.6 +/- 107.2 mg/day). The patients were studied with Holter ECG monitoring (Schiller MT-100, Switzerland) and high resolution ECG (electrocardioanalyser Cardis, Geolink-electronics, RF) with construction of periodograms of ff waves and interval histograms RR (IHrr), estimation of the rhythm variability (SDRR, rMSSD, PNN50). RESULTS: Beta-blockers (atenolol, metoprolol), verapamil and diltiazem had no significant effect on the period of ff waves. The degree of a mean heart rate lowering decreased in the following order: beta-blockers-verapamil-diltiazem (30.1 +/- 12.5, 25.0 +/- 18.8 and 22.0 +/- 23.6 beat/min, differences are insignificant), this corresponded to the degree of Rrmin increase (0.12 +/- 0.04, 0.08 +/- 0.07 and 0.07-0.08). In CCF the inhibiting effect of beta-blockers and verapamil is substrate-dependent: the shorter baseline Rrmin (and higher heart rate), the more potent is the effect due to action of beta-blockers and verapamil (r = -0.58 and r = -0.57, p < 0.05) and reduction of a mean heart rate (r = -0.74 and r = -0.84, p < 0.05). Dependence of diltiazem effect on initial Rrmin is inverse. In contrast to Ca antagonists (verapamil, diltiazem), beta-blockers increased latent conduction manifesting in a significant rise of Rrmax, range of RR intervals (the difference between Rrmax and Rrmin) and in increased latent conduction (Rrmax/Rrmin) by 0.4 versus 0 and 0.1 in the groups on verapamil and diltiazem). In addition to insufficient shift RRmod, there appeared non-optimal rhythm structure--combination of a large number of short and long RR in small number of middle ones. Verapamil and diltiazem improved the rhythm pattern due to proportional increase of RRmin shift RRmod (r = 0.72 and r = 0.71, p < 0.05) and absence of a distinct effect on latent conduction. The between groups differences by SDRR, RMSSD and PNN50 dynamics were insignificant. Diltiazem in doses 360-480 mg/ day moderately increased latent conduction, but was low effective in the presence of early peak RR (0.28-0.46 s). CONCLUSION: Monotherapy with AB-blocking drugs was possible only in patients with moderate tachycardia, no waves of fibrillation of large and middle periods (0.15 s and higher) and should be conducted under Rrmin control. In the other cases, the above drugs are either low effective or promote non-optimal rhythm structure. Therefore, combined therapy with AB-blocking drugs and cardiac glycosides is indicated for CCF patients. PMID- 17078215 TI - [Effects of atorvastatin and acipimox on the lipid spectrum of blood plasma, endothelial function and a clinical course of unstable angina pectoris]. AB - AIM: Assessment of effects of a 3-month course of atorvastatin and a nicotinic acid derivative acipimox on the lipid spectrum, platelet aggregation, results of brachial artery reactive hyperemia test and clinical outcomes in patients with unstable angina pectoris (UAP). MATERIAL AND METHODS: A total of 109 males with new-onset and progressive UAP entered the trial. They had LDLP cholesterol > or = 115 mg/dl. RESULTS: The addition of atorvastatin to combined therapy of UAP patients resulted in achievement of a target level of LDLP cholesterol level in 64% dose-adjusted patients while in acipimox patients the target level was achieved in 20% patients. In spite of a more potent effect on endothelial function, acipimox had an insignificant effect on the treatment outcomes. CONCLUSION: Hypolipidemic therapy with atorvastatin and acipimox for UAP leads to a significant reduction in LDLP cholesterol. The effect of atorvastatin comes much earlier and more frequently provides a target level of LDLP cholesterol. PMID- 17078216 TI - [New approaches to treatment of bradyarrhythmia in patients with obstructive sleep apnea syndrome]. AB - AIM: To confirm or reject effects of CPAP on sleep asystole and to elucidate possible dependence on correction of respiratory disorders. MATERIAL AND METHODS: Thirteen patients (11 males and 2 females aged 19 to 66 years) with bradyarrhythmia (BA) arising in sleep participated in the study. BA was caused by transient atrioventricular block of the degree II-III in 9 cases (69%), episodes of sinus node arrest and/or synoatrial block in 6 (46%) cases. Two (15%) patients had combination of BA forms. To diagnose sleep respiratory disorders, polysomnography (PSG) was made. Diagnostic criteria of sleep obstructive apnea syndrome (SOAS) were apnea/hypopnea index (AHI) more than 5 episodes for 1 hour sleep. The patients were divided into two groups. Individual selection of therapeutic pressure under PSG control was performed in SOAS patients (the study group). CPAP-therapy was effective in AHI < 5. In the control group (AHI < 5) such selection was made too. CPAP-therapy was effective in the controls if episodes of apnea/hypopnea were not registered through the night of monitoring. RESULTS: In the study group CPAP-therapy was effective. The AHI decreased from 73.2 to 4.4, oxygen saturation of arterial blood increased from 74 to 85%, mean duration of asystoles fell from 5.2 to 1.3 s, pauses with duration more than 2 s disappeared. In the control group sleep apnea/hypopnea episodes disappeared but in asystole CPAP was uneffective. CONCLUSION: CPAP-therapy is effective and pathogenetically sound method of treating patients with nocturnal bradyarrhythmia associated with sleep respiratory disorders. PMID- 17078217 TI - [Cardiomyoplasty with autological mononuclear cells of the bone marrow in patients with acute myocardial infarction]. AB - AIM: To study efficacy and safety of transplantation of bone marrow autologous mononuclear cells (BMAMC) in patients with acute myocardial infarction; to examine BMAMC distribution in the human body after intracoronary introduction. MATERIAL AND METHODS: The open controlled trial investigated 26 AMI patients (16 entered the study group and 10 were controls). Cell cardiomyoplasty with BMAMC was performed by intracoronary injection of the cells after stenting the coronary artery supplying blood to the infarction zone on AMI day 7-21. BMAMC were isolated by gradient centrifugation. Distribution of mononuclear cells was studied with radionuclear indication of the cells 99m-Tc-HMPAO. All the patients were examined with Tl-199 perfusion scintigraphy of the heart 2 weeks and 6 months after the treatment, echocardiography, 24-h ECG monitoring, 6-min walk test. RESULTS: All the patients were followed up for 6 months. Two patients (one in each group) developed recurrent myocardial infarction 3 months after the first. Radionuclide investigations revealed fixation of labelled mononuclear cells in the heart both in initial hours after the treatment and 24 hours after it. As shown by myocardial scintigraphy, intracoronary administration of the cells with short-term arterial occlusion was followed by much greater number of labeled cells. By follow-up month 6, in the study group, left ventricular ejection fraction increased more: 12.7 +/- 3.2% versus 10.4 +/- 2.5% in the control group (p = 0.09); moreover, a stable defect of myocardial perfusion reduced more (by 29 +/- 24% against 20 +/- 18%, respectively, p = 0.1). Malignant arrhythmia, complications during and after bone marrow aspiration, intracoronary administration of cell suspension were not registered. CONCLUSION: Intracoronary administration of BMAMC in AMI patients is safe and provides their transfer and fixation in the myocardium. BMAMC transplantation has a positive effect on recovery of perfusion and contractile function of left ventricular myocardium in AMI patients. PMID- 17078218 TI - [Splenectomy in programmed therapy of aplastic anemia]. AB - AIM: To define efficacy of splenectomy (SE) in current programmed therapy of aplastic anemia (AA). MATERIALS AND METHODS: SE efficacy was analysed in 2 stages: a retrospective study of efficacy of surgery as monotherapy (1986-1996) (74 AA patients) and of SE in programs of immunosuppressive therapy (IST) (1991 2002). Program treatment of AA patients was conducted on the base of IST algorithm developed in Hematological Research Center after many year investigations. RESULTS: SE as monotherapy improved AA course in 73.3% patients with non-severe AA (NAA) and 18.2% patients with severe AA (SAA). Three and five year survival in NAA postsplenectomy patients was 80%. One-year survivors after surgery were likely to survive long. Overall survival of SAA after SE was significantly less (p < 0.0001): 3-year survival - 6%. SE efficacy in programs including antilymphocytic globulin (ALG) and cyclosporin A (CsA) was studied in 69 AA patients. A 85.5% response was registered to program treatment including ALG, CsA and SE, being 81% in SAA and 1% in NAA patients. Efficacy of SE in combination with CsA at the first stage NAA treatment (a 30% positive response) was much inferior to ALG+CsA (68% response). At stage two treatment SE improved treatment results in most of SAA patients. Long-term survival in SAA patients after program treatment with SE is 60%. CONCLUSION: SE in the program of combined therapy in adult AA patients including CsA is an alternative to ALG in NAA patients. In SAA, SE can be included in the program at the first stage in ALG intolerance or in the absence of the drug, at the second stage--to overcome resistance to conducted therapy. PMID- 17078219 TI - [Ten-year outcomes of lymphogranulomatosis treatment according to the protocol MOPP-ABVD+radiotherapy]. AB - AIM: To analyse overall recurrence-free survival of lymphogranulomatosis (LGM) patients given polychemotherapy (PCT) MOPP (mustargen-caryolisin, vincristine, natulan, prednisolone) - ABVD (adriamycin, bleomycin, vinblastin, dacarbasin) in combination with radiotherapy (RT) for 10 years. MATERIAL AND METHODS: The trial included 211 LGM patients admitted to Hematological Research Center in 1990-1996 from other hospitals without random selection. The patients were examined by the standard program including biopsy of the affected organ or lymph node, bilateral trephine biopsy. Splenectomy was performed in 17 patients, 83 patients received PCT in other hospitals, 128 untreated patients received MOPP-ABVD therapy (3 courses of MOPP and 3 courses of ABVD). Forty one patients had defects in PCT, 16 of them rejected PCT and RT. The latter was performed 4 weeks after the 6th course, contraceptives were not prescribed to women. At LGM stage II-III RT was performed by the subradical program (no radiation to ilioinguinal lymph nodes) in doses 40-44 Gy on the foci and 32-36 Gy preventively, on massive and residual foci after PCT - 5-10 Gy additionally. RESULTS: Ten-year overall and recurrence free survival in the untreated group reached 83 and 80%, respectively, for pretreated patients - 46 and 36%, respectively. Causes of death of 26 patients were LGM progression, infection (tuberculosis, as a rule), secondary tumors and acute myeloblastic leukemia (AML). After remission 25 women gave birth to a healthy child and 12 healthy children were born to 9 males. CONCLUSION: MOPP-ABVD plus radiotherapy program according to subradical and radical variants was in the past effective but invalidating rescue therapy. Present-day programs consider the histological variant, stage and prognostic factors allowing an individual therapeutic approach with step-by-step reduction of RT in the treatment of LGM patients. Involvement of the bone marrow in primary patients had no influence on the treatment results. This refers this affection not to a generalized stage IV, but to stage III along with involvement of the lymph nodes and the spleen. PMID- 17078220 TI - [The role of telmisartan in the treatment of metabolic syndrome]. AB - AIM: To evaluate telmisartan effects on 24-h blood pressure profile, sensitivity to insulin, carbohydrate and lipid metabolism in patients with metabolic syndrome (MS) and arterial hypertension (AH). MATERIAL AND METHODS: Twenty patients with MS and AH of the degree I-II received telmisartan in a dose 80 mg/day. 24-h BP monitoring, tests for total cholesterol, HDLP, LDLP cholesterol, triglycerides, glucose, glucose tolerance test were made before and 24 weeks after the treatment. RESULTS: Telmisartan reduced all the study parameters of blood pressure, body mass, fasting and post-prandial hyperglycemia, postprandial hyperinsulinemia. Telmisartan raised peripheral tissue sensitivity to insulin, normalized phases of insulin secretion. Total cholesterol, LDLP cholesterol diminished while HDLP went up. CONCLUSION: Telmisartan has a specific ability to partially activate receptors stimulating proliferation of peroxisomes and improve regulation of carbohydrate and lipid metabolism, to reduce body mass. PMID- 17078221 TI - [Lipid reducing and antioxidant action of mexicor in patients with diabetes mellitus type 2]. AB - AIM: To study mexicor effects on functional activity of beta-cells, insulin resistance, lipid metabolism and lipid peroxidation in patients with diabetes mellitus (DM) type 2. MATERIAL AND METHODS: Twenty patients with DM type 2 participated in a double blind randomized trial of mexicor vs placebo. Before and after therapy the following parameters were studied: plasma glucose before meal, immunoreactive insulin, glycosilated hemoglobin, cholesterol, triglycerides, LDLP and HDLP cholesterol, malonic dialdehyde, dienic conjugates, superoxide dismutase, catalase, glutathione peroxidase and alpha-tocopherol. RESULTS: Mexicor significantly improved compensation of carbohydrate metabolism by glucose and glycosylated hemoglobin in the blood, insulin resistance value, lipid metabolism and lipid peroxidation in activation of antioxidant enzymes. CONCLUSION: Mexicor in therapy of DM type 2 improves carbohydrate and lipid metabolism, lipid peroxidation, activates antioxidant defence enzymes, functional activity of beta-cells, reduces insulin resistance. PMID- 17078222 TI - [Efficacy of vaccine therapy with ribomunil in patients with chronic obstructive pulmonary disease]. AB - AIM: To assess efficacy of vaccine therapy with ribomunil in prevention of exacerbations and prolongation of remission in patients with obstructive pulmonary disease (COPD). MATERIAL AND METHODS: Fifty three COPD patients were divided into two groups: 28 patients of the study group received ribomunil plus standard therapy; 25 controls received only standard treatment. The examination of the patients included blood count, sputum cytology, external respiration function test, immunological investigation (CD4+, CD8+, CD20+, CD25+ and CD95+; IL-4, IFN-gamma, complement). The examination was made before the treatment, 21 days, 3 and 6 months after its beginning. Ribomunil was given 3 tablets in the morning before meal for 4 days in a week for 3 weeks, 4 days each month for the following 5 months. RESULTS: As a result of ribomunil treatment lymphocyte subpopulation normalized, immune response shifted to Th-1 dependent immune response, external respiration flow indices improved, remission prolonged. CONCLUSION: Ribomunil is recommended for wide application in the practice of COPD treatment PMID- 17078223 TI - [Immunogenetic analysis of variants of a clinical course and prognosis of chronic glomerulonephritis in the west Siberia]. AB - AIM: To analyze association of HLA antigens with clinical variants of chronic glomerulonephritis (CGN)in 106 patients. MATERIAL AND METHODS: HLA association with the time of development of terminal chronic renal failure (CRF) from the moment of detection of initial CRF was studied in 80 patients. Determination of HLA of class I was conducted in the lymphocytotoxic test. DRB1 genes specificity was determined with DNA polymerase chain reaction. The control group consisted of 341 healthy citizens of Novosibirsk. Common HLA associations of latent and hypertensive, latent and nephrotic CGN variants were found demonstrating their succession. HLA associations with a mixed CGN variant defining its genetic isolation were established. It is shown that development of terminal CRF at different time since the disease onset is associated with different HLA alleles. The findings allow prognosis of clinical CGN variants, time of development of terminal CRF. PMID- 17078224 TI - [Treatment and prevention of critical states in cardiovascular diseases]. PMID- 17078225 TI - [Is the role of thiazine-like diuretics verified in therapy of arterial hypertension?]. PMID- 17078226 TI - [Efficacy of a new ACE inhibitor zofenopril (zocardis) in cardiovascular diseases]. PMID- 17078228 TI - Called to excellence. PMID- 17078227 TI - [Tiklopidin--30 years of clinical application]. PMID- 17078229 TI - Rx for compassion fatigue. PMID- 17078230 TI - Magnet recognition: the Rush experience. PMID- 17078231 TI - Magnet nursing and Kingdom of God principles. PMID- 17078232 TI - Principles for a healthy work environment. PMID- 17078234 TI - When it's time to leave. PMID- 17078233 TI - TV nurses often hurt more than heal. PMID- 17078235 TI - Healthcare: one body, many parts. PMID- 17078236 TI - Whose outcome is it? PMID- 17078237 TI - The other side of the bed. PMID- 17078238 TI - The day Max died. PMID- 17078239 TI - Words that heal. PMID- 17078240 TI - After Katrina nurses take action. PMID- 17078241 TI - Encountering discrimination. PMID- 17078242 TI - Can we pray with students? PMID- 17078243 TI - "You're welcome". PMID- 17078244 TI - Time for spiritual care? PMID- 17078245 TI - Postgraduate specialization: an alternative to undergraduate streaming and limited licensure. PMID- 17078246 TI - An ethicist's commentary on substituting cremains. PMID- 17078247 TI - Seroprevalence of Mycobacterium avium subspecies paratuberculosis, Neospora caninum, Bovine leukemia virus, and Bovine viral diarrhea virus infection among dairy cattle and herds in Alberta and agroecological risk factors associated with seropositivity. AB - A province-wide, cross-sectional seroprevalence and agroecological risk factor study of Mycobacterium avium subspecies paratuberculosis (MAP), Neospora caninum (NC), Bovine leukemia virus (BLV), and Bovine viral diarrhea virus (BVDv) genotypes 1 and 2 (BVDv1 and BVDv2) infection in dairy cattle herds in Alberta was conducted. Among adults, the seroprevalence of MAP, NC, and BLV was 9.1%, 18.5%, and 26.9%, respectively. For MAP, based on a herd test cutpoint of 2 or more seropositive cows, 58.8% of herds were infected. Herd-level seroprevalence for NC and BLV was 98.7% and 86.7%, respectively, based on a herd-test cutpoint of 1 seropositive cow. Among unvaccinated dairy heifers, the seroprevalence for BVDv1 and BVDv2 infection was 28.4% and 8.9%, respectively, while herd-level infection was 53.4% and 19.7%. Seroprevalence for MAP varied moderately by agroecological region, whereas that for NC, BLV, and BVDv1 and BVDv2 did not. For MAP, aridity and soil pH (correlated features of the region) were also important. PMID- 17078248 TI - Clinical characteristics and predictors of mortality for Cryptococcus gattii infection in dogs and cats of southwestern British Columbia. AB - Since 1999, Cryptococcus gattii has emerged as an important pathogen of humans and animals in southwestern British Columbia. Historically thought to be restricted to the tropics and subtropics, C. gattii has posed new diagnostic and treatment challenges to veterinary practitioners working within the recently identified endemic region. Clinical reports of canine and feline cryptococcosis caused by C. gattii diagnosed between January 1999 and December 2003 were included in this case series. The most common manifestations of disease were respiratory and central nervous system signs. Multivariate survival analysis revealed that the only significant predictor of mortality was the presence of central nervous system signs upon presentation or during therapy. Case fatality rates in both species were high. Further investigation into effective treatment regimes is warranted. PMID- 17078249 TI - Efficacy of orbifloxacin tablets for the treatment of superficial and deep pyoderma due to Staphylococcus intermedius infection in dogs. AB - Orbifloxacin tablets were administered orally to 23 dogs with superficial and/or deep staphylococcal pyoderma. Response to therapy was excellent in 95.6% of the dogs. Duration of therapy varied from 21 to 40 days (average 29 days) for dogs having only superficial infections, and from 25 to 150 days (average 72 days) for dogs having deep infections. Relapses occurred in 18% of the dogs within a 3 month period. One dog developed a presumed adverse cutaneous drug reaction. Under the conditions of this study, orbifloxacin was an effective, safe, and convenient antibiotic for the treatment of superficial and deep staphylococcal pyoderma in dogs. PMID- 17078250 TI - Segmental tracheal dysplasia in a mixed breed dog. AB - A 6-month-old, spayed female, mixed breed dog was referred for respiratory difficulty. Radiographic and bronchoscopic evaluations showed tracheal narrowing from the thoracic inlet to its bifurcation. Under anesthesia, the trachea ruptured and the dog died. Microscopic evaluation revealed underlying segmental tracheal and bronchial cartilaginous malformation (dysplasia). PMID- 17078251 TI - Septic epiglottic chondritis with abscessation in 2 young Thoroughbred racehorses. AB - Septic epiglottic chondritis with abscessation diagnosed in 2 Thoroughbred racehorses. Infected cartilage removed videoendoscopically followed by systemic antibiotics. The infectious process was successfully controlled, but permanent dorsal displacement of the soft palate (DDSP) with a shortened, deformed epiglottic cartilage developed. Surgery for the DDSP using bilateral partial sternothyroidectomy or laryngeal tie-forward failed. PMID- 17078252 TI - Isolation of a Mycobacterium microti-like organism from a rock hyrax (Procavia capensis) in a Canadian zoo. AB - A Mycobacterium tuberculosis complex organism was isolated from a zoo resident rock hyrax (Procavia capensis) imported into Canada from South Africa. The strain was identified biochemically as Mycobacterium microti. The spoligotype pattern obtained for this isolate was found to be rare. This represents the first report of isolation and spoligotyping of M. microti in North America. PMID- 17078253 TI - Diagnosis and successful treatment of Cryptococcus neoformans variety grubii in a domestic ferret. AB - A domestic ferret was presented for episodic regurgitation. Cytologic examination and culture of an enlarged submandibular lymph node revealed Cryptococcus neoformans variety grubii (serotype A). The ferret was successfully treated with itraconazole. This is the first documented case of Cryptococcus neoformans variety grubii in a ferret in the United States. PMID- 17078255 TI - Problematic client-veterinarian relationships: the "yes, buts". PMID- 17078254 TI - Chronic pneumonia and polyarthritis syndrome in a feedlot calf. AB - A feedlot heifer was diagnosed with chronic pneumonia and polyarthritis syndrome (CPPS), which was confirmed on postmortem examination. The syndrome is caused by Mycoplasma bovis; however, available data support the theory that other pathogens play a role in predisposing to this disease. Continued research on CPPS focuses on pathogenesis, treatment, and prevention. PMID- 17078256 TI - Veterinary diagnostic imaging. Retroperitoneal abscess and regional cellulitis secondary to a pyelonephritis within the left kidney. PMID- 17078257 TI - Patellar luxation: pathogenesis and surgical correction. PMID- 17078258 TI - [Inhospital prognosis of acute myocardial infarction: comparison of two periods to ten years of interval]. AB - Acute myocardial infarction prognosis depends on many factors relative to the patient, to the disease and to the quality and promptness of the treatment. The interventional revascularisation is superior to pharmacological revascularisation. The aim of the study is to compare acute myocardial infarction inhospital lethality in the coronary care unit before and after use of emergency angioplasty. The retrospective study compared inhospital lethality over two four years periods separated by an interval of ten years: [1991-1994] and [2001-2004]. Lethality had increased from 7.8% to 11.3%. During the second period emergency angioplasty (under 24 hours) primary (4.8%) or rescue (2.5%), not applied in the nineties, was not related to survival and did not allow an improvement of patients' hospital prognosis. The population of the second period was characterized by: a greater average age (60.8 vs. 58.6 years), more female subjects (20.1 vs. 14.6%), less nicotinic subjects (60.2 vs. 78.5%) and less patients receiving fibrinolytic treatment (43.0 vs. 50.4%). These parameters were as many factors related to the risk of hospital death in univariate analysis. The multivariate analysis showed that after adjustment on all the other factors the lack of thrombolysis remains unquestionably related to the hospital overlethality. The time delays for admission in the intensive care unit were relatively long and comparable over the two periods (8.2 vs. 7.5 hours). Within our hospital structure early angioplasty not being able to be realized in the hour, which follows the diagnosis on a large scale, and in a continual way, it would be preferable to always consider thrombolysis first in all cases apart from contraindications without excluding the possibility of a hybrid treatment. PMID- 17078259 TI - [Cognitive impairment induced by digoxin intake in patients older than 65 years]. AB - Cognitive impairment or clinical signs of dementia in an old patient who receives digoxin, should suggest a digitalis intoxication. Symptoms can be present although a normal digoxin serum concentration. It is recommended to stop the treatment to obtain a regression of dementia symptoms. PMID- 17078260 TI - Heart rate variability in rats with chronic hypoxic pulmonary hypertension. AB - The precise role of pulmonary hypertension as a possible factor inducing a decrease in heart rate variability is poorly known. Spectral analysis of heart rate variability (HRV) was carried out in 21 Wistar rats before and after exposure to normoxia (N = 10) or to 3 weeks of hypobaric hypoxia inducing chronic pulmonary hypertension and right ventricular hypertrophy (N= 11). Continuous ECG was recorded in conscious animal at rest. Compared to the control group, rats exposed to hypoxia had a similar heart rate but a lower overall HRV (total power, 27.9 +/- 15.2 vs. 57.6 +/- 24.7 ms2, P < 0.01). Low frequency power (0.25-0.8 Hz) and high frequency power (0.8-3 Hz) were similar in both groups suggesting that HRV was decreased in the very low frequency power (0-0.25 Hz). The effects of atropine and propranolol on heart rate and HRV were similar in rats exposed or not to hypoxia. HRV is decreased in rats with hypoxic induced pulmonary hypertension, mainly in the very low frequency band, suggesting an increase in sympathetic activity. However, this decrease is moderate and the modulation of HRV with pharmacologic autonomic blockade remains similar to that of normal rats. PMID- 17078261 TI - [Cardiac sequelae of penetrating cardiac wounds]. AB - OBJECTIVE: The aim of our work was to study cardiac sequelae of penetrating cardiac injuries. MATERIAL AND METHODS: It is a retrospective study including nine patients (eight men and one woman with an average age of 26.6 +/- 7.4 years) hospitalised between January 1st, 1988 and December 31, 2002 in cardiovascular surgery department of Tunis Rabta hospital for heart wounds. RESULTS: It was stab wounds in all patients. Heart lesions concerned primarily the right ventricle. In admission, two patients had cardiac tamponade and seven were in shock. Beating heart surgery without cardiopulmonary bypass in all patients performed surgical repair and it consisted in simple suture. Functional symptoms were observed in five patients at late outcome. Five patients had heart sequelae. It consisted of conduction abnormalities (right bundle branch block) in five patients. One patient had a small ventricular septal defect. Finally, partial permanent disability rates were less than 10%. CONCLUSION: Cardiac sequelae of penetrating cardiac wounds have good prognosis with the exception of a small partial permanent disability rate. Their prevention must be based on the fight against the urban violence and on the good initial management. PMID- 17078262 TI - [The left ventricular reconstructive surgery at the origin of the treatment of cardiac insufficiency]. AB - The surgical method of ventricular reconstruction described by Dor is recalled with the clinical report of a patient who presented a ventricular aneurysm. The left ventricular reconstructive surgery is based on an anatomical design of the heart described by Torrent-Guasp, where the normal orientation of the left ventricular muscle fibers, oblique in direction, is found parallel with the base of the heart at the time of ventricular dilation. By giving again an elliptic form to the left ventricle, the left ventricular reconstructive surgery improves the cardiac function of the patient who developed a bulky aneurysm after an infarction. Based on this concept, other techniques of ventricular reconstruction intended for patients presenting dilated cardiomyopathy, of ischemic origin or not, are being studied. PMID- 17078263 TI - [The role of B type natriuretic peptide in the assessment of post myocardial infarction prognosis]. AB - Recently cardiac peptides have received close attention as cardiovascular markers. Brain (B type) natriuretic peptide is a neurohormone synthesized predominantly in ventricular myocardium. Previous studies have shown that this hormone can provide prognostic information in patients with myocardial infarction. The aim of this review is to evaluate the impact of plasma levels of BNP on prediction of left ventricular ejection fraction and remodelling and major cardiac events after myocardial infarction. PMID- 17078264 TI - [Peripartum cardiomyopathy]. AB - Peripartum cardiomyopathy is a rare and under recognized form of dilated cardiomyopathy, defined as a heart failure in the last month of pregnancy or in the first five months post-partum with absence of determinable cause for cardiac failure and absence of demonstrable heart disease. The incidence of peripartum cardiomyopathy ranges from 1 in 1300 to 1 in 15,000 pregnancy. Advanced maternal age, multiparity, twin births, preeclampsia and black race are known risk factors. The etiology of peripartum cardiomyopathy remains unknown but viral, autoimmune or idiopathic myocarditis are highly suggested. The clinical presentation on patients with peripartum cardiomyopathy is similar to that of patients with systolic heart failure. The treatment is based on drugs for sympyomatic control. Studies in graeter populations are need to determine the role of immunosupressive treatment. About half patients of peripartum cardiomyopathy recover. The left ventricular ejection fraction and the left ventricular end-diastolic diameter are statistically significant prognostic factors. The risk of developing peripartum cardiomyopathy in subsequent pregnancies remains high. The place of dobutamine stress test in counseling the patients who desire pregnancy must be more studied. PMID- 17078265 TI - [Clinical experiences with the new electromagnetic ventricular assist devices]. AB - Recently, the third generation of partial electromagnetic artificial hearts appeared, which are a real technological advancement. These new ventricular assist devices are small, implantable, silent and increase the patients' quality of life in comparison with the electromechanical pump of second generation. Their clinical efficiency is at least as good as the second generation in light of the first experiences reported on animals and human beings. Their use as a bridge to transplant and especially as a destination therapy should increase in the coming years as they represent an answer to the health problem, which is the increasing population of patients in heart failure. PMID- 17078266 TI - [Osborn J wave. A new "channel pathology"? A case report]. AB - We report, at the time of a hypothermia major, the observation of an anomaly of the repolarisation on the electrocardiogram of surface, called "J wave", and described in an exhaustive way by Osborn, which attached its name there. It corresponds to the picking of the terminal section of the QRS, with heightening in dome, the J point is then elevated compared to the base line. It can be also seen among patients normothermic in physiological or pathological circumstances. Its physiopathology from now on is understood better, the J wave is the result of the difference of potential action between the epicarde and endocarde during phases 1 and 2 of the ventricular repolarisation. This gradient is related to the Ito current, also accused in the "channel pathologies", of which Brugada syndrome. PMID- 17078267 TI - [Beneficial effect of sildenafil following surgery for mitral stenosis complicated by pre-capillary pulmonary hypertension: report of two cases]. AB - Pulmonary hypertension is a serious disorder, difficult to treat especially in the severe forms. The treatment consists mainly of calcium channel blockers, anti coagulation, intravenous epoprostenol, inhaled nitric oxide and recent agents as bosentan and sildenafil. Sildenafil, a phosphodiesterase 5 specific inhibitor, has been largely evaluated in primary pulmonary hypertension, and in some cases of secondary pulmonary hypertension including parenchymal and thromboembolic diseases; it has not yet been evaluated in severe pulmonary hypertension with elevated pre-capillary resistance in operated mitral stenosis. We report the cases of two patients operated from mitral valve replacement for severe mitral stenosis with elevated pre-capillary resistance, where oral sildenafil, introduced empirically immediately after the surgical procedure at the dose of 50 mg/d, permitted a significant decrease in pulmonary pressures and resistances, allowing a rapid withdrawal of nitric oxide and reducing therefore hospitalization time in the intensive care unit. We think that this simple treatment, with or without association to nitric oxide, should be generalized to persistent pulmonary hypertension following cardiac surgery. PMID- 17078268 TI - [Superior mesenteric artery aneurysm revealing a Behcet disease: a case report]. AB - Arterial involvement in Behcet's disease is rare. It represents one of the fashions of expression of this pathology. We report an observation of a 32-year old man who was hospitalised for an intense abdominal pain. Doppler ultrasound and computed angiography showed a superior mesenteric artery aneurysm. The patient was operated in urgency and operating suites were simple. The etiologic inquiry of this aneurysm ended in the diagnosis of Behcet disease. The result underline the need to search for Behcet's disease in all cases of arterial involvement in young patient. The treatment of the aneurysm should not be delayed. PMID- 17078269 TI - [Anastomotic false aneurysms after under-diaphragmatic bypasses: about 25 cases]. AB - The purpose of our study is to determine the causes and the management of anastomotic aneurysms. We report the cases of 25 patients with a mean age of 64 years at the time of initial surgical revascularisation. The mean interval between the first operation and the occurrence of anastomotic aneurysm is 5 years (range 2 months-11 years). The treatment consists on the interposition of a graft in 8 patients, anastomotic angioplasty in 9 patients and the reconstruction of the anastomosis in 8 patients. Hospital mortality was 20%. Late death occurs 3 patients and the long-term morbidity was evaluated at 22%. A recurrence of anastomotic false aneurysm occurred in 4 patients (16%) (Range 7 months-1 year). In conclusion anastomotic false aneurysm is one of the major complications of vascular reconstruction; careful follow-up can detect the rare instances of anastomotic aneurysm and reoperation can be accomplished with a low-rate of adverse outcome. PMID- 17078270 TI - [Thoracic aortic dissection revealed by systemic cholesterol embolism]. AB - Systemic cholesterol embolism is a rare complication of atherosclerosis, and has various presentations. Arterial catheterisms are a common cause. However, the association with an aortic dissection has been exceptionally reported. We report the observation of a 70 year-old man, with coronary artery disease, hypertension, diabetes and dyslipidemia. Six months before hospitalization, a coronary angioplasty was performed due to recurrent angina. The association of purpuric lesions on the feet, with acute renal failure confirmed cholesterol embolism syndrome. Transoesophageal echocardiography showed a dissection of the descending thoracic aorta associated with complex atheroma. The evolution was marked by the pulpar necrosis of a toe and by a worsening of the renal failure, requiring definitive hemodialysis. Further echographic control highlighted the rupture of the intimal veil of the dissection. Cholesterol embolism syndrome may reveal an aortic dissection in patients without thoracic symptoms. In such cases, transoesophageal echocardiography is a useful and non-invasive examination. PMID- 17078271 TI - Obesity and periodontal disease. PMID- 17078272 TI - A conservative approach for the management of a crown-root fracture. AB - Subgingival crown-root fractures are major complications that jeopardize tooth survival and require the clinician to adopt a multidisciplinary approach combining different dental specialties to effectively treat the teeth and ensure long-term survival. Fragment reattachment is a conservative therapeutic option for uncomplicated crown-root fractures. However, biologic width involvement must be carefully evaluated and addressed. This article describes a case of restoration of a crown-root fracture by means of periodontal surgical exposure of the fracture, crown and root recontouring, and fragment reattachment. PMID- 17078273 TI - Systemic diseases among patients with and without alveolar bone loss. AB - OBJECTIVE: Growing evidence shows an association between periodontal and general health. This study was undertaken to compare the prevalence of hypertension, respiratory diseases, diabetes, arthritis, and allergies among patients with and without alveolar bone loss. METHOD AND MATERIALS: A total of 420 individuals aged 18 years and older were selected. Self-reported systemic diseases, age, gender, marital status, smoking, alveolar bone level, and number of teeth present were obtained from the subjects' dental records prior to treatment. Chi-square, Student t test, and logistic regression were used for data analysis. RESULTS: Hypertension, respiratory diseases, diabetes, and arthritis were significantly more prevalent among subjects with moderate to severe alveolar bone loss. Hypertension and diabetes remained significantly more prevalent among subjects with more severe alveolar bone loss even after adjusting for confounders. Also, subjects with moderate to severe alveolar bone loss were 4 times more likely than their counterparts to have 3 or more systemic conditions. CONCLUSION: Subjects with moderate to severe alveolar bone loss have an increased prevalence of systemic diseases, especially hypertension and diabetes mellitus. PMID- 17078274 TI - Correlation between maxillary central incisor form and face form in 4 racial groups. AB - OBJECTIVES: The purpose of this study was to evaluate the correlation between central incisor form and face form in 4 racial groups and to investigate if there was agreement among experts in categorizing the central incisor forms. METHOD AND MATERIALS: A total of 160 subjects (40 whites, 40 mulattos, 40 blacks, and 40 Asians) ranging from 18 to 33 years of age were selected. Digital photographic records were made, 1 full-face and 1 intraoral view of the maxillary right central incisor. The outline tracings of the tooth and face images were obtained using Adobe Photoshop 5.0 software. The outline tracings were printed in distinct transparencies, and 3 prosthodontists determined if there was correspondence between the tooth and the face forms by superimposition of the transparencies. If there was disagreement among the prosthodontists' evaluations, the prevalent decision was considered. The experts also classified the central incisor forms into square, ovoid, tapering, or combination at 2 different sessions. At the first session, no instructions were given. At the second session, the prosthodontists were instructed to follow Williams' method of classification. RESULTS: A correspondence between tooth and face forms was found in 23.75% of all cases. Agreement on the tooth form classifications among the prosthodontists occurred in 30.62% of all cases at the first session and 24.37% at the second session. CONCLUSION: There is not a highly defined correlation between central incisor form and face form in any racial group studied. In addition, the experts were not in fair agreement in categorizing tooth forms. PMID- 17078275 TI - Deep impacted mandibular second molar: a case report. AB - Impaction of mandibular second molars is relatively rare. A case of a deep impacted mandibular right permanent second molar in a 26-year-old woman is presented. This article describes the extraction of the tooth and reports the risks of the surgical procedure. Careful manipulation of tissues and correct application of mechanical principles involved in tooth extractions allowed surgical success. PMID- 17078276 TI - Dental care of patients with autoimmune vesiculobullous diseases: case reports and literature review. AB - Dental management of patients with autoimmune vesiculobullous disorders is complicated because of prominent involvement of oral mucosa, increased risk of oral disease, and difficulty in rendering dental care. Although these diseases are relatively uncommon, dental practitioners should be familiar with the oral sequelae of these conditions and their management. Pemphigus vulgaris, cicatricial pemphigoid, and epidermolysis bullosa represent the most common autoimmune oral vesiculobullous diseases. This case-illustrated review summarizes the pathogenesis, diagnostic features, and natural history of oral vesiculobullous disorders, placing an emphasis on the treatment and prevention of associated oral disease aimed at maintaining a healthy, functional dentition. PMID- 17078277 TI - Changing dietary behavior. PMID- 17078278 TI - Efficacy of a computerized shade selection system in matching the shade of anterior metal-ceramic crowns--a pilot study. AB - OBJECTIVE: The purpose of this study was to determine whether anterior crowns fabricated using a computerized shade selection system (ShadeScan, Cynovad) (experimental procedure) match adjacent teeth better than anterior crowns fabricated using conventional shade prescription and clinical slides (control). METHOD AND MATERIALS: Five subjects who required a crown to restore a maxillary central incisor were selected. Two metal-ceramic crowns were fabricated for each incisor, 1 using the experimental procedure and 1 using the control method. The shade selection method to be used for the first and second crowns was randomly assigned. The duration of each procedure was recorded. Each restoration was tried in in a double-blind manner and evaluated for its level of match to adjacent teeth using modified Ryge criteria. Data were analyzed within each subject using descriptive statistics and paired t test (alpha = .05). RESULTS: In 40% of the cases, both procedures did equally well. In the remaining 60% of the cases the control procedure (two-thirds of the cases) performed better than the experimental procedure (one-third of the cases). Duration of the control procedure was 14.4 +/- 5 minutes, and the experimental procedure was 5.2 +/- 3.3 minutes. A paired t test showed the difference was significant (P = .0045). CONCLUSION: The level of matching of crowns fabricated with the ShadeScan system was not different from crowns fabricated using the control. However, it took significantly less time to record the shade with the ShadeScan system. PMID- 17078279 TI - Voids and interlayer gaps in Class 1 posterior composite restorations: a comparison between a microlayer and a 2-layer technique. AB - OBJECTIVE: The main objective was to compare the presence of interlayer gaps between 2 hybrid composites placed using a microlayer and a 2-layer technique. METHOD AND MATERIALS: Standardized Class 1 cavities were prepared in 40 extracted posterior teeth. Two resin composite materials were used. The control group, group A, consisted of samples of the materials extruded out of the manufacturers' syringes. Group B consisted of 10 teeth restored using 2 layers per restoration. Group C consisted of 30 teeth restored using 6 microlayers per restoration. A scanning electron microscope was used to detect voids in the samples. RESULTS: Round, well-defined voids were found in 85% to 100% of the samples within the bulks of the resin composite materials only. A statistically significant relationship between the type of composite and presence of bubbles (Fisher exact test, P = .04) was found among the 2-layer technique group. No statistically significant difference (Fisher exact test, P = .48) was found among the microlayer technique group. Irregular interlayer gaps were found in 5% to 15% of specimens within areas between bulks of composite, as detected by an electron microscope based on the different densities of the bulks of composite and the interlayer areas. No such statistically significant relationship was found (Fisher exact, P = .62) among the study groups. CONCLUSIONS: Voids are routinely found in bulks of composite materials. Higher incidences of interlayer gaps in the 2-layer samples suggest that the use of a microlayering technique may result in fewer gaps. PMID- 17078280 TI - Comparison of K hand files and ProFiles 0.06/0.04 in simulated curved root canals prepared by students. AB - OBJECTIVES: The objective of this study was to compare the step-down technique in simulated curved root canals using K hand files (VDW) and ProFiles 0.06/0.04 (Dentsply Maillefer). METHOD AND MATERIALS: A total of 72 canals were blindly prepared by instructed students: 1 block with K files and a second one with ProFiles 0.06/0.04 in connection with the EndoStepper (SET). Postoperative images of the canals were taken, and an independent dentist assessed preparations for amount and position of material removed during preparation. Efficacy of the instruments is described in terms of canal shape, instrument failure, canal blockages, and preparation time. RESULTS: ProFile instrumentation provided well centered and tapered preparations while maintaining the apical foramen; in contrast, the use of K files resulted in frequent alterations of the root canal. In the apical portion, both techniques showed unprepared areas. The coronal diameter in hand-instrumented canals was more irregular and wider than in those mechanically prepared. There was no separation of a ProFile instrument. Hand instrumentation was faster than mechanical, but the difference was not statistically significant. CONCLUSION: Under the conditions of the study, the ProFile rotary system was simple and safe to use and created better preparations than hand instrumentation. PMID- 17078281 TI - Esthetic management of the upper labial frenum: a novel frenectomy technique. AB - An abnormal upper labial frenum is capable of retracting the gingival margin, creating a diastema, limiting lip movement, and in cases of a high smile line, affecting esthetics also. When an abnormal frenum is present, frenectomy is advised. This technique has been modified in many ways. But in most of the techniques the zone of attached gingiva and esthetics are not considered. This case report highlights a new technique of frenectomy that results in good esthetics, excellent color match, gain in attached gingiva, and healing by primary intention at the site of the thick, extensive abnormal frenum. PMID- 17078282 TI - [Drinking water supply in the Russian Federation]. PMID- 17078283 TI - [Hygienic characteristics of foodstuffs containing genetically modified components]. AB - The paper analyzes the results of the investigations of raw foods, foodstuffs for genetically modified components, conducted by the state sanitary and epidemiological service of the Russian Federation during its current sanitary inspection. The presented materials cover 2003-2004. The findings suggest that there is a great deal of foods containing genetically modified sources on the market and show the priority groups of foodstuffs and the distribution of these foods on the territory of the Russian Federation. PMID- 17078284 TI - [Role of hygienic studies in Moscow residents' health support]. AB - The Government of Moscow allocates substantial funds for the development of a sociohygienic monitoring (SGM) system and for the guidance of the population's health status monitoring in relation to environmental factors. The risk-assessing studies made jointly with the investigators of the A. N. Sysin Research Institute of Human Ecology and Environmental Hygiene, Russian Academy of Medical Sciences, make it possible to determine priorities in risk control and to introduce new technologies for controlling the public health associated with the influence of social and environmental factors. PMID- 17078285 TI - [Investigation of the bactericidal effectiveness of ultraviolet irradiation in different operating modes of recirculatory irradiators]. PMID- 17078286 TI - [Hygienic evaluation of water supply in the oil-producing areas of the Republic of Tatarstan]. AB - The paper gives hygienic characteristics of water supplying conditions in the oil producing areas of the Republic of Tatarstan. A long-term intensive oil-field development, without taking into account its environmental influence, is shown to have resulted in the pollution of water resources and broken the conditions of water consumption by the population. There has been a considerable increase in the total mineralization (1.8-2.3 g/l) of rivers, the levels of chlorides (300 1200 mg/l) and sulfates (300-1800 mg/l), and the salinity of springs. The implementation of nature-conserving and sanitary-and-hygienic measures have stabilized and improved the qualitative composition of water sources used for drinking water supply in the oil-producing areas of the Republic of Tatarstan, leading to a marked trend for improving water supplying conditions and accordingly the quality of the population's life. PMID- 17078287 TI - [Results of biomedical monitoring in the elimination of iodine deficiency in the Tyumen Region]. PMID- 17078288 TI - [The evaluation of contamination of water objects with hepatitis A virus (HAV) and the impact of HAV on morbidity trends in large seaports of the Ukraine]. AB - The authors studied changes in the contamination of urban water objects with hepatitis A virus (HAV) (over 11 years) and analyzed its association with HAV infection morbidity in Odessa residents. The study showed that HAV antigen was isolated in 5.5% of all cases. The seawater was maximally contaminated (6.93%), the water of the Dniestr River and that of the distribution network were less contaminated--3.9 and 4.9%, respectively. The maximum quantity of HAV-containing drinking water samples were found in 1994 (52.6%) and 2002 (11.7%). In 1994-2004, there was a decrease in the contamination of drinking water with HAV antigen from 52.6% to 0.59%, which correlated to the reduction in morbidity rates (r = 0.73; p < 0.05). PMID- 17078289 TI - [Ambient air benz[a]pyrene and cancer morbidity in Kemerovo]. AB - A statistically significant direct strong correlation was found between the annual average daily concentrations of air benz[a]pyrene and the lung and the gastric cancer morbidity rates in males and females, skin, thyroid, and ovarian cancer in females. The certain interval of the measured concentration of benz[a]pyrene and the recorded morbidity rate was shown to be characteristic of each of the above-mentioned tumors. PMID- 17078290 TI - [Hygienic assessment of carcinogenic and noncarcinogenic risks of oral intake of drinking water chemical agents]. AB - Based on the quantitative and qualitative determination of drinking water pollution by a large number of substances of varying toxicity and hazard, the authors calculated a possible risks of their adverse impact on the body by the development of carcinogenic and noncarcinogenic effects. PMID- 17078291 TI - [Hygienic evaluation of air ionization in public premises: analytical aspects]. PMID- 17078292 TI - [Characterization of the sanitary situation in the Kurgan Region]. AB - The aim of this survey was to study the ecological state in the Kurgan Region in the period of 1993 to 2003. The survey showed the west and northwest part of the Kurgan Region to be areas at risk for environment-dependent diseases. The major risk factors included a high pollution of water sources with limited water resources due to the radioactive pollution occurring in the 1940s to the 1950s in some areas of the region; transformation of natural landscapes caused by spring floods and soil erosions among other things. PMID- 17078293 TI - [Agricultural use of fertilizers: ecological and hygienic aspects]. AB - The paper considers the topical aspects of safe agricultural use of fertilizers. Temporary hygienic standards for the fertilizer levels of toxic elements have been developed: 8 mg/kg for cadmium; 15 mg/kg for lead; 10 mg/kg for arsenic. PMID- 17078294 TI - [Correlation between the probability and dosage characteristics of safety in the processing of radioactive liquids]. AB - The authors show it possible and expedient to use the probability characteristics for evaluation of the safe operation of radiation-dangerous plants. The calculated values of doses that might be following an accident at these plants fail to give an adequate account of reality since the doses are largely variates. To divide accidents into classes or levels, it is necessary to use the principle of non-excess of accident probability of the given class or level rather than the threshold principle of non-excess of doses. PMID- 17078295 TI - [Assessment of results of interlaboratory comparative tests for food toxic elements]. PMID- 17078296 TI - [Optimization of analytical methods for assay of ochratoxin A in the foodstuffs]. AB - Ochratoxin A is a mycotoxin produced by widespread mold fungi of the genera Aspergillius and Penicillium. It is a common contaminant of cereals, coffee, wine, dried fruits, and spicery. There is evidence that ochratoxin A has nephrotoxic, immunosuppressive, embryotoxic, teratogenic, and carcinogenic effects on many species of mammals. Its maximum allowable concentration established by the Alimentarius Code and the European Union is 5 microg/kg for cereals and 3 microg/kg for ready-made cereals. Two optimized techniques (silica gel column chromatography (CC) or immunoaffinity CC for extract purification and high performance liquid chromatography with fluorescence detection) are proposed for the detection, identification, and assay of ochratoxin A in food commodities. The detection limit was 0.05 and 0.5 microg/kg, respectively. The developed techniques were used to assay the content of ochratoxin A in 46 samples of the grain gathered in different regions of Russia. Nine samples were contaminated by ochratoxin A at concentrations of 0.05 to 5 microg/kg. PMID- 17078297 TI - [Modern military school education sociomedical analysis]. AB - The sociomedical status of senior military school students was studied, by examining the formed stereotype of a healthy lifestyle, spiritual and patriotic values, as well as physical health indices. It was established that among the significant spiritual and patriotic values, most (62.3%) cadets reported love for their country in the first three places: 68.4% of the schoolchildren called physical health; 47.6% put a readiness at defending the motherland in the first three places while 53.9% named physical health. Psychoemotional studies revealed a low level of anxiety and neurotization in the cadets than in the schoolchildren. Physical health studies (hand muscle strength, vital capacity, hypoxic resistance) indicated that the senior military school students had higher fitness than the general educational school children. These differences in the physical health indices of the adolescents are chiefly associated with the greater attention given by the cadets to their physical education than that shown by the general educational schoolchildren. PMID- 17078298 TI - [Morphofunctional features of the body's development in primary-form pupils]. AB - An experiment was carried out to study the functioning of the basic systems in 350 primary-form pupils from a multiartistic lyceum, a multidisciplinary gymnasium, and a traditional school during 3-year study. The time course of changes in the cardiorespiratory system suggests its higher stresses in of children from innovational schools. Overwork and inactivity are prominent among the school factors that influence the children's health. PMID- 17078299 TI - [Age-related changes in the psychophysiological parameters in male teenagers]. AB - The time course of changes in the time of a simple visuaI motor reaction, force and lability of the nervous system, stability of attention and memory was studied in 2104 urban and rural males aged 18-27 years to develop standard phychophysiological status rating tables. It was shown that the score of the central nervous system should be used to detect some functional impairments of the central nervous system. Positive changes in higher nervous performance were noted, which were more pronounced in young rural males. Rating scales were developed to diagnose the individual characteristics of the central nervous system in young men of call-up age. PMID- 17078300 TI - [The physical development and psychophysiological status of children born to mothers engaged in gas-chemical industry]. AB - The study has ascertained that the adaptive capacities of the body reflect the level of its health and the resistance to the influence of environmental factors. The organism of a child and his/her functional capacities of the basic systems may be regarded as an indicator of the quality of life in a mother exposed to poor factors of gas-chemical industry; and there is likely to be a long-term impact of gas-processing factors on the child through the maternal body. PMID- 17078301 TI - [Smoking in the able-bodied population: spread and sociodemographic characteristics]. PMID- 17078302 TI - [Characterization of the biological properties of acid-treated chrysotile asbestos fibers]. AB - Boiling of chrysotile of the textile brand PRZh1-50 in concentrated hydrochloric acid for 10, 15, and 20 minutes gave rise to three chrysotile-asbestos samples. The content of MgO decreased from to 24, 19, and 9%, respectively. As compared with the baseline values, the number and force of positively charged electrical centers were less in the samples containing 24 and 19% MgO and more in the sample having 9% MgO; the negatively charged centers were present in the former two samples and absent in the third one. When the samples were intrapleurally administered to rats, their hemolytic activity, induction of active oxygen radicals, mutagenic activity (micronuclear test using murine bone marrow cells), and the frequency of mesotheliomas were less in the treated samples than in the baseline ones; but there were no differences between the treated samples. Thus, the altered physicochemical properties of the fibrillar surface of asbestos diminished its biological aggressiveness; however, increased treatment rates failed to lead to its further decrease. There was no relationship of the biological properties to the number and force of electric charges of the surface. PMID- 17078303 TI - [Comparative evaluation of the safety of poly- and and oligoguanidines]. AB - The antimicrobial activity and microenvironmental safety of poly- and oligoguanidine antiseptics were compared. E. coli and St. aureus test strains were found to be more sensitive to chlorohexidine bigluconate (CHB) upon a long (24 hour) exposure and to polyhexamethylene guanidine derivatives on short (30 second) contact. It is concluded that unlike polyguanidine antiseptics, CHB is more ecologically dangerous; when used for rapid disinfection, it can provoke impairment in the microenvironmental balance outside the area of application. The findings show it expedient to assess the microenvironmental safety of antimicrobial agents in order to prevent ecological catastrophes. PMID- 17078304 TI - [Effect of licorice (Glycyrrhiza) on the activity of succinate dehydrogenase upon exposure to vibration]. PMID- 17078305 TI - [State sanitary and epidemiological surveillance of individual respiratory organ protectors at radiation-dangerous establishments]. PMID- 17078306 TI - [Design of an experiment studying the dose-time dependence of an effect of a combination of adverse factors]. PMID- 17078307 TI - [Impact of the personality characteristics of railway assemblers on occupational traumatism]. PMID- 17078308 TI - [An assay of food sulfamethoxazole residues]. PMID- 17078309 TI - The effects of nicotine on the metabolic and hormonal responses during acute cold exposure. AB - OBJECTIVE: To examine the effects of nicotine on the metabolic and hormonal responses during acute cold exposure. METHODS: Participants in this study included 6 men and 5 women between the ages of 19 and 25 years. Each subject performed 2 cold-air trials (CATs) consisting of a 30-minute baseline (BASE) period and a 120-minute exposure to 10 degree C air. One CAT was performed after a nicotine (NIC) dosing using a 21-mg transdermal patch, whereas the other CAT was performed after a placebo (PL) treatment. Blood samples for metabolic and hormonal measurements were obtained at the end of BASE and immediately after the cold exposure. RESULTS: When examining the sexes separately, there was no difference in norepinephrine between PL and NIC (P = .066). There was also no difference in epinephrine between PL and NIC in either sex (P = .634). From BASE to 120 minutes of the CAT, there was a significant decrease in cortisol (P = .036), but this response was similar between the 2 treatments (P = .077). Glucose and glycerol concentrations were not different between the PL and NIC treatments. At BASE, nonesterified fatty acid (NEFA) concentration was lower during PL compared with NIC (P = .021); however, at 120 minutes of the CAT, NEFA was greater during PL compared with NIC (P = .035). CONCLUSIONS: During 120 minutes of cold exposure, NIC resulted in alterations in the responses in NEFA, whereas the other blood measurements were not significantly different between the 2 groups. PMID- 17078310 TI - Pattern of injury and illness during expedition-length adventure races. AB - OBJECTIVE: To describe injuries and illnesses treated during an expedition-length adventure race and combine the results with those from previous studies to identify common patterns of injury and illness during these events. METHODS: The 2003 Subaru Primal Quest Expedition Length Adventure Race was held in Lake Tahoe, CA, from September 5 to 14, 2003. Eighty teams of 4 individuals participated. During the event, medical volunteers providing on-site medical care recorded each medical encounter on a medical encounter form. This information was used to describe the injuries and illnesses treated and was combined with previous investigations to identify common patterns of injury and illness during these events. RESULTS: During the 10-day study period, 356 patient encounters and 406 injuries and illnesses were recorded. The most frequent reason to require on-site medical care was injury of the skin and soft tissue (70.4%), with blisters the single most common of these injuries (45.6%). Other reasons were orthopedic injury (14.8%), respiratory illness (3.7%), and heat illness or dehydration (3.7%). CONCLUSIONS: The results of this and previous studies demonstrate a common pattern of injury and illness that includes a high frequency of skin and soft tissue injury, especially blisters. Injuries and illnesses such as altitude illness, contact dermatitis, and respiratory illness varied considerably among events. The number of patient encounters per athlete is similar among the studies, providing an approximation of the number of medical encounters expected given the number of participants. These results should assist medical providers for future events; however, it is imperative to carefully review the individual event to best predict the frequency of injury and illness. PMID- 17078311 TI - Back pain in whitewater rafting guides. AB - OBJECTIVE: The purpose of this study was to quantify rates of back pain among whitewater rafting guides and to look for correlations between the presence of back pain and specific activities associated with guiding. The secondary objective was to provide suggestions for outfitters according to the results of this study, which may be of assistance to their guide staff. METHODS: A mail-out mail-back survey was sent to 2510 rafting guides working in Washington, Oregon, Idaho, Utah, Colorado, and the Grand Canyon during the summer of 2004 to quantify whitewater raft guiding characteristics and incidence of back pain. RESULTS: Of the 390 surveys returned, 77.4% of guides reported back pain while guiding and 20.8% had back pain lasting longer than 1 week at the time of the survey. Stacking 5 or more inflated boats for transport was correlated with the presence of pain (chi(2) = 8.4, v = 1, P < .01), and loading and unloading rafts while guests are waiting was correlated with back pain lasting longer than 1 week (chi(2) = 8.1, v = 1, P < .01). CONCLUSIONS: The rates of back pain among, and activities of, whitewater rafting guides were reported. Rates of back pain among whitewater rafting guides who returned our survey appear similar to the general population. Although determining a particular cause of pain is difficult, the typical injury seems to be relatively minor in scope. PMID- 17078312 TI - Changes in metabolic and hematologic laboratory values with ascent to altitude and the development of acute mountain sickness in Nepalese pilgrims. AB - OBJECTIVE: During August of each year, thousands of Nepalese religious pilgrims ascend from 2050 m to 4500 m in 1 to 3 days. Our objectives were to evaluate the incidence of acute mountain sickness (AMS) among this large group of native people, to explore changes in serum electrolytes as subjects ascend to high altitude, and to attempt to determine whether decreased effective circulating volume is associated with the development of AMS. METHODS: This was a prospective study with 2 parts. In the first part, demographic, physiologic, and laboratory data were collected from a cohort of 34 pilgrims at both moderate (2050 m) and high altitude (4500 m). Changes that occurred with ascent were compared in subjects who did and did not develop AMS. The second part was a cross sectional study of a different group of 57 pilgrims at the high-altitude site to further determine variables associated with AMS. RESULTS: In the cohort of 34 subjects, Lake Louise score, heart rate, respiratory rate, blood urea nitrogen (BUN), BUN:creatinine ratio, and pH increased at high altitude, whereas oxygen saturation, bicarbonate, creatinine, and PCO2 decreased. Sixteen of these 34 subjects (42%) were diagnosed with AMS; these patients had a statistically significantly lower hematocrit, oxygen saturation, and self-reported water consumption than those without AMS. Of the 57 subjects enrolled in the cross sectional study, 31 (54%) were diagnosed with AMS. These pilgrims had higher heart rates and BUNs than did their non-AMS counterparts. CONCLUSIONS: Fifty-two percent of the subjects developed AMS. With ascent to altitude, subjects showed some evidence of decreased effective circulating volume, though there were no clinically significant changes. The data did not show whether decreased circulating volume is a significant risk factor in the development of AMS at high altitude. PMID- 17078313 TI - Digital amputation caused by climbing-rope entanglement. AB - Digital avulsions and amputations are not uncommon in outdoor activities. Rope entanglement is often the cause, though specific descriptions of the mechanism of injury have not been reported. We report a common scenario where entanglement during a fall resulted in an amputation injury. Although any roping activity may cause a loop formation leading to digital entrapment and subsequent amputation injury, activities involving soft and loose ropes may increase the chances of injury. Rope slack is to be avoided to minimize the likelihood of entanglement. This case report describes a digital amputation during an indoor climbing incident and also describes a possible mechanism for such injuries. PMID- 17078314 TI - Prolonged urinary incontinence and biliary dyskinesia following abdominal contact with jellyfish tentacles. AB - A 16-year-old girl was seriously stung on her abdomen by a jellyfish as she jumped on her small surfboard. She and her mother identified the animal from photographs as Chrysaora fuscescens. Within several minutes the girl developed a massive abdominal cutaneous eruption composed of hundreds of punctuate erythematous papules and macules, which persisted for 5 to 7 days. Persistent urinary incontinence and biliary dyskinesia appeared over the following night. It is theorized that a systemic uptake of venom occurred percutaneously after contact of the jellyfish tentacles with her abdominal skin. The result was an injury to the urinary and biliary bladders. This is the first case report of such sequellae after topical contact with a marine animal. The causal relationship of these abnormalities with the sting is suggested by their temporal association. The gallbladder disorder required surgical intervention, but spontaneous resolution of the urinary bladder dysfunction occurred within 20 months. PMID- 17078315 TI - Injuries at the 2005 World Championships in Rock Climbing. AB - OBJECTIVE: To evaluate the injury risk associated with indoor rock climbing competition. METHODS: All injuries reported to medical personnel at the 2005 World Championships in Rock Climbing were recorded and analyzed. RESULTS: Four hundred forty-three climbers (273 men, 170 women) from 55 countries participated in 3 separate disciplines totaling 520 climbing days. Only 4 of 18 acute medical problems that were treated were significant injuries, resulting in an injury rate of 3.1 per 1000 hours. CONCLUSIONS: Indoor rock climbing competition has a low injury risk and a very good safety profile. PMID- 17078316 TI - Spinal injuries in Scottish mountaineers. AB - OBJECTIVE: To identify the frequency and pattern of spinal injury sustained in mountaineering accidents. METHODS: All patients with spinal injuries sustained while mountaineering who were treated at the Queen Elizabeth National Spinal Injuries Unit from 1992 to 2001 were studied. Information was obtained from hospital notes, and each patient completed a questionnaire. RESULTS: Twenty-one patients with spinal injuries sustained in mountaineering accidents were identified. Fourteen patients (67.7%) were experienced mountaineers. Four (19.0%) were rock climbing, 6 (28.6%) were winter climbing, 6 (28.6%) were hill walking, and 5 (23.8%) were winter walking. Ten patients (47.6%) sustained cervical injuries, 5 (23.8%) sustained thoracic injuries, 5 (23.8%) sustained lumbar injuries, and 1 (4.8%) sustained cervical and lumbar injuries. Nineteen patients (90.5%) sustained fractures, 1 (4.8%) sustained a dislocation, and 1 (4.8%) sustained a fracture dislocation. Seventeen patients (81.0%) were neurologically intact, 2 (9.5%) sustained complete cord injury, and 2 (9.5%) sustained incomplete cauda equina lesions. Ten patients (47.6%) required internal fixation of their fractures. Fourteen patients (57.1%) sustained other significant injuries. CONCLUSIONS: This is the first study specifically aimed to describe patterns of spinal injuries and their outcomes in mountaineers. The group we studied had sustained fractures at differing spinal levels with no specific injury pattern. The incidence of cord injury was lower than in other spinal injury patient groups. The majority had also sustained other significant and potentially distracting injuries. Prehospital care providers should maintain a high suspicion of spinal injury in this group. PMID- 17078317 TI - Goats and gases: "The Prevention of Compressed Air Illness" by Haldane et al--A commentary. PMID- 17078318 TI - Painful fissures and diabetes mellitus at altitude. PMID- 17078319 TI - Nonbite exposures to rattlesnakes: a retrospective review from a statewide poison control system. PMID- 17078320 TI - High-altitude pulmonary edema at moderate altitude as first manifestation of pulmonary hypertension in a 14-year-old boy with Down Syndrome. PMID- 17078321 TI - Teaching and learning in urology nursing. PMID- 17078322 TI - An overview of adult-learning processes. AB - Part of being an effective instructor involves understanding how adults learn best. Theories of adult education are based on valuing the prior learning and experience of adults. Adult learners have different learning styles which must be assessed prior to initiating any educational session. Health care providers can maximize teaching moments by incorporating specific adult-learning principles and learning styles into their teaching strategies. PMID- 17078323 TI - Effective teaching strategies for the older adult with urologic concerns. AB - Older adults are able to learn and apply new knowledge, such as management of a urological condition, but their learning style differs from that of younger clients. An understanding of the older adult's learning style, and knowledge of specific teaching strategies will benefit the urologic nurse in the ongoing effort to be a more effective patient educator. PMID- 17078324 TI - Using Peplau's theory of interpersonal relations to guide the education of patients undergoing urinary diversion. AB - Patients diagnosed with bladder cancer may require a urinary diversion to maximize their health care outcomes. These patients, faced with sudden changes in their health status, develop complex unmet needs that can be addressed by a planned program of education. Peplau's theory of interpersonal relations offers a framework for patient teaching that emphasizes the importance of the nurse patient relationship. This therapeutic relationship enables the nurse to provide the patient with the information needed to understand the diagnosis, cooperate in the treatment plan, facilitate postoperative recovery, and return to a state of independence with quality of life. PMID- 17078325 TI - Engendering student empathy for disabled clients with urinary incontinence through experiential learning. AB - As part of a rehabilitation clinical course for senior baccalaureate nursing students, a disability-incontinence experiential learning activity is required. The assignment is intended to familiarize students with some of the challenges encountered by a client with mobility problems, including continence management issues using disposable undergarments. Wearing the undergarments dry and wet while being confined to a wheelchair provides insight and promotes empathy for patients with bladder control problems. PMID- 17078326 TI - Games as an innovative teaching strategy for overactive bladder and BPH. AB - A challenge for urologic nurses and nurse educators is how to present information to staff, students, and patients in a way that will capture their interest and engage them in the learning process. The use of adult-learning principles and innovative teaching strategies can make the learning experience dynamic, and encourage learners to take a more active role in their own learning. Games are a creative, fun, and interactive way to assist in the emphasis, review, reinforcement, and retention of information for urology nurses. PMID- 17078327 TI - Bringing Mohammed to the mountain: educating the community for continence. AB - The Bladder Health Mobile is an innovative community education initiative developed by the Simon Foundation for Continence. It is intended to provide education, increase public awareness, and promote early diagnosis and proper treatment of incontinence and other bladder control problems, while also facilitating dialogue between consumers and their health care professionals; the hope is to alleviate the stigma associated with bladder conditions. PMID- 17078328 TI - Cultural disparities in the diagnosis and treatment of prostate cancer. PMID- 17078329 TI - Continuing education changes affect urologic nursing journal. PMID- 17078330 TI - Getting ready for certification: obstructive uropathy. PMID- 17078331 TI - Implantable LHRH agonist for treatment of prostate cancer. PMID- 17078332 TI - The transparent organization: leadership in an open organization. AB - The impersonal approach to health care leadership is over. Specialization, hierarchies, and impersonal decisions have led the public to distrust health care organizations. The charges of unnecessary cardiac surgery and abuses led the public to question our integrity. Annison and Wilford (1998) note that the character of a person and the character of an organization lead one to trust or distrust. They note that openness is one of the most important characteristics upon which we judge the character of a person or an organization. As the operating framework of partnerships and transparency becomes one that our staff and patients expect, our ability to handle openness will be an important way in which we are judged. PMID- 17078333 TI - Update on infectious risks of blood transfusions. PMID- 17078334 TI - Cholesterol, cholesterol-lowering agents/statins, and urologic disease: Part IV- is vitamin E phosphate or another unique type of this vitamin the last chance for these supplements? AB - Vitamin E, once the highly espoused heart healthy paradigm of the dietary supplement world, has now been touted by many experts to be either heart unhealthy or simply ineffective based on the results of past and recent randomized trials in cardiovascular medicine. Natural and synthetic vitamin E supplements, especially in large dosages, have been the concern. Researchers have embarked on testing a variety of other novel forms of vitamin E, such as vitamin E phosphate and other forms of vitamin E, to determine safety issues as well as the impact on cardiovascular disease (CVD) risk markers and end-points. Until vitamin E demonstrates meaningful changes in some of the accepted markers of CVD prevention, it will be difficult to espouse their use specifically for these and other conditions. PMID- 17078335 TI - Air Force Dental Corps aims to stem shortage. PMID- 17078336 TI - The bisphosphonates struggle. PMID- 17078337 TI - Michigan Supreme Court clarifies peer review immunity. PMID- 17078338 TI - Dealing with Internet use and abuse. PMID- 17078339 TI - Adjustments: The hidden overhead. PMID- 17078340 TI - Why CareCredit works: the proof is in the numbers. PMID- 17078341 TI - Guidelines for management of patients with periodontal disease. PMID- 17078342 TI - Your official planning guide to "give kids a smile" day. PMID- 17078343 TI - A question of professionalism: are we treating patients as people or procedures? PMID- 17078344 TI - Top 10 tips for buying a dental practice (that you'll love). PMID- 17078345 TI - Evidence: what is it and why do we need it? PMID- 17078346 TI - Implementing the role of patient-navigator nurse at a university hospital centre. AB - A profile of the role and functions of an oncology patient-navigator nurse (OPN) and the preliminary phases to implementing this role within a team specializing in oncology are first presented. This is followed by a qualitative study that provides a descriptive assessment for implementing an initial OPN in the head and neck oncology area of a university hospital centre (UHC) with a supraregional model for oncology. Three groups of stakeholders (individuals with cancer and families, caregivers, network partners) were interviewed on three occasions: before, during and after implementation. The results show that this new role can be integrated within a team specializing in oncology. The beneficial effects of this role on the process of adaptation to illness, interdisciplinary work and continuity of care are described. Several recommendations are formulated, one being the importance of situating the implementation process from an organizational change perspective. PMID- 17078347 TI - Development of an oral care guide for patients undergoing autologous stem cell transplantation. AB - Nurses identified oral mucositis as a recurring issue in clinical practice. To meet this challenge, a group of nurses took a leadership role in developing an oral care guide. The University Health Network Nursing Research Utilization Model and the Neuman Systems Model served as conceptual frameworks. A flowchart was developed to ensure a coordinated and continuous provision of oral care. Educational presentations were conducted to familiarize nurses and members of the multidisciplinary team of the practice changes. The introduction of the oral care regimen as primary prevention, plus systematic oral assessment and monitoring had the potential to reduce the occurrence and severity of oral mucositis in patients undergoing autologous stem cell transplantation. PMID- 17078348 TI - Understanding the symptoms experienced by individuals with lung cancer. AB - The purpose of this study was to gain a better understanding of patients' experience of symptoms of lung cancer upon a first visit to a regional cancer centre and two months later. Sixteen patients were interviewed on two occasions about the symptoms, their impact and what they found most troublesome. Patients were also asked to describe any strategies they used to manage the symptoms. The most commonly identified symptoms were fatigue and pain. Participants described coping with multiple symptoms simultaneously, how those symptoms intensified over time, and using very few strategies. Participants described symptoms as troublesome because they interfered with activities of daily living or evoked emotional or cognitive responses. Fatigue was reported most frequently as troublesome. This work has implications for how patient assessments are conducted and how health care professionals listen to the patients. The patients' descriptions of their symptoms and what strategies they applied is often embedded within the patients' stories about living day-to-day with their lung cancer. PMID- 17078349 TI - Touched by a nurse: the imprint of exemplary oncology nursing care. AB - This paper examines the relationship between exemplary oncology nursing practice and professional fulfillment. What inspires nurses to choose oncology and, subsequently, what gives them impetus to continue in this field? To answer this question, oncology nurses were invited to recall moments in their careers when they were satisfied that they had chosen the "right" career. Data, in narrative form, were collected through an online research technique. Submissions were analysed using three approaches, narrative analysis (Priest, Roberts, & Woods, 2003), poetic interpretation (van Manen, 1990) and photovoice (Woolrych, 2004). Findings reveal that oncology nurses who provide excellent care, and make strong connections with their patients, are also usually very satisfied with their careers. Specifically, nurses provide exemplary care and report attaining professional fulfillment when they achieve connection with those in their care by affirming value and sharing humour. Second, caregivers feel they are making a difference when they "see patients through" the care trajectory. Nurses accomplish this in part by helping people live on, individualizing care, enabling hope, and helping individuals find meaning. It is anticipated that this paper will reawaken memories of similar experiences in caregivers, thus enhancing confidence, self-esteem and energy and reminding nurses that they do unquestionably leave an imprint. PMID- 17078350 TI - Philosophy at the table with nursing--the interface of transdisciplines in psychosocial oncology. PMID- 17078351 TI - Evaluate business plans, select strategic partners by taking a 10-step approach. PMID- 17078352 TI - The advent of the consumer-patient. PMID- 17078353 TI - Embracing leadership. PMID- 17078354 TI - The time has come. Describing and shaping RN practice. PMID- 17078355 TI - RN practice during a pandemic... How ready are you? PMID- 17078356 TI - The nurse practitioner: a new setting. PMID- 17078357 TI - Breaking down racial barriers. Honouring pioneer Aboriginal nurses from the Blood Reserve. PMID- 17078358 TI - Adventures in nursing. PMID- 17078359 TI - Health by the people. PMID- 17078360 TI - Like the energizer bunny, these retirees keep going and going. PMID- 17078361 TI - Biomeds work in shadow of nation's capital. PMID- 17078362 TI - Improving patient safety: more than repairs for today's biomeds. PMID- 17078363 TI - Interplay between IT and CE benefits patients, hospitals. PMID- 17078364 TI - The legacy of the CE and BMET: safer healthcare technologies. PMID- 17078365 TI - Pulse oximeters 101. PMID- 17078366 TI - Acoustic stethoscopes. PMID- 17078367 TI - Looking for trouble: proactive troubleshooting. PMID- 17078368 TI - JCAHO warns of tubing misconnects; communication is key. PMID- 17078369 TI - What is Health Level 7? AB - HL7 is a huge undertaking and continues to grow. We talked about certain main themes of the spec that I've found interesting and thought you might too. You can always find additional information-especially from the HL7 website itself at www.hl7.org or www.altisinc.com/ IE/hl7.html. The main thing to remember is that HL7 is a specification for describing, formatting, encoding, and sharing of clinical and administrative data in healthcare. An easy way to remember that is that the 7 in HL7 refers to the Application layer in the OSI model. The next time someone talks about their HL7 software or application, you'll know that they really mean their HL7 compliant software or application. Then be sure to ask them how many composites are in their OBX segments! PMID- 17078370 TI - Time to talk salary, but when? PMID- 17078371 TI - Preparing for the certification exam. PMID- 17078372 TI - Monthly reporting process empowers staff and strengthens teamwork. PMID- 17078373 TI - Recall management: best practice basics. PMID- 17078374 TI - Risk management for IEC 60601-1 third edition. AB - With the examples above we have demonstrated for our selected topics that our device meets the risk management requirements of 60601-1. An expansion of the tables to include all of the risk management requirements would be useful to identify how the manufacturer has met all of the risk management requirements of the standard. By setting up a file system under the manufacturer's quality system that meets the requirements of the 14971 risk management standard, the manufacturer will be able to effectively demonstrate compliance with a number of applicable standards, including 60601-1, and also be able to demonstrate the device meets FDA product safety requirements. The file system would also be useful in creating the technical file for the MDD. A risk management summary table similar to the one recommended by the GHTF will assist the manufacturer in tracking the documentation required to support this effort. A list of the hazards identified for the product is contained in the summary table and the summary table will also aid in determining if all of the required steps in the risk management process have been completed for each hazard. This table will become an important record in the risk management file. PMID- 17078375 TI - The tao of managing recalls and safety alerts. AB - Clinical engineering staff deal with a variety of risk management issues on a daily basis. These issues range from infection control, standards compliance, recalls and safety alerts, patient safety, and incident investigations all the way up to disaster preparedness. Managing device recalls and safety alerts entails several functions, such as processing, risk assessment, distribution, rectification, tracking, and monitoring. This paper discusses the basic elements of an effective recalls and safety alerts management system, thus offering the way to an effective system. The system enabling inputs, activities, and desired outcomes are discussed. The paper also presents our experience in implementing such a system and future possibilities. PMID- 17078377 TI - Time to reboot: new software standard to replace SW68. PMID- 17078376 TI - Simulation in medicine: addressing patient safety and improving the interface between healthcare providers and medical technology. AB - Medicine, as an industry in which human lives depend on the skill and performance of operators, must create and maintain a culture of safety, in addition to promoting the design of systems to mitigate errors. The use of medical simulation as a mechanism for training healthcare professionals in a safe environment is expanding rapidly. An important component of systems that ensure the safety of patients in the hospital setting is the interface between humans and technology in the hospital. The objective of this paper is to review: (1) the definition and a brief history of medical simulation, (2) examples of how current medical simulation centers are using simulation to address patient safety, and (3) examples of how simulation can be used to enhance patient safety through improvement of the interface between healthcare practitioners and medical technology. Medical simulation and human factors engineering can be used to examine and enhance the interface between healthcare practitioners and medical technology, with the potential to make a significant contribution to patient safety. PMID- 17078378 TI - Why can't biomed be more like IS? PMID- 17078379 TI - The ethics of gene therapy. AB - Recent developments have progressed in areas of science that pertain to gene therapy and its ethical implications. This review discusses the current state of therapeutic gene technologies, including stem cell therapies and genetic modification, and identifies ethical issues of concern in relation to the science of gene therapy and its application, including the ethics of embryonic stem cell research and therapeutic cloning, the risks associated with gene therapy, and the ethics of clinical research in developing new therapeutic technologies. Additionally, ethical issues relating to genetic modification itself are considered: the significance of the human genome, the distinction between therapy and enhancement, and concerns regarding gene therapy as a eugenic practice. PMID- 17078380 TI - Assessing and communicating the risks and benefits of gene transfer clinical trials. AB - The two most important ethical requirements for clinical studies of gene transfer are to ensure that: (i) a protocol's risks are minimized and acceptable in light of the potential benefit to society and to research participants; and that (ii) a valid, informed consent process takes place. The history of gene transfer studies in humans shows that the probability of harm or benefit to participating individuals is extremely low. However, because the risks and benefits of gene transfer remain unpredictable, and because increasing the potential for benefits to individuals is likely to be accompanied by increases in potential for adverse events, close monitoring of human gene transfer research will continue. Recent research on informed consent for phase I studies of serious illnesses has revealed an evolving discussion on the proper standards for disclosure; one important conclusion is that disclosures regarding potential benefits to individuals need to be less ambiguous than they currently are. PMID- 17078381 TI - Development of hematopoietic stem cell gene therapy for Wiskott-Aldrich syndrome. AB - Wiskott-Aldrich syndrome (WAS) is a complex primary immunodeficiency disorder associated with microthrombocytopenia, autoinnmunity and susceptibility to malignant lymphoma. At the molecular level, this rare disorder is caused by mutations in the gene encoding the Wiskott-Aldrich syndrome protein (WASP). WASP is a cytosolic adaptor protein mediating the rearrangement of the actin cytoskeleton upon surface receptor signaling. Allogenic hematopoietic stem cell (HSC) transplantation represents a curative approach but remains problematic in light of severe risks and side effects. Recently, HSC gene therapy has emerged as an alternative treatment option. Cumulative preclinical data obtained from WASP deficient murine models and human cells indicate a marked improvement of the impaired cellular and immunological phenotypes associated with WASP deficiency. The first clinical trial is currently being conducted to assess the feasibility, toxicity, and potential therapeutic benefit of transplanting autologous WASP reconstituted hematopoietic stem cells. PMID- 17078382 TI - Stem cell therapy after myocardial infarction: ready for clinical application? AB - The discovery of stem cells capable of generating angiogenic or contractile cells and structures might offer new treatment options for patients suffering from heart disease. In particular, embryonic stem cells are considered to have great potential for regenerative medicine and tissue engineering. Studies suggest that delivery or mobilization of stem and progenitor cells might improve tissue perfusion and contractile performance of the damaged heart; however, the underlying mechanisms are poorly understood. Fusion or trans-differentiation into cardiomyocytes or vascular cells are considered rare events of cellular engraftment, and adult stem cells are now considered as 'regenerator cells', acting via paracrine effects of cytokines, or by activation of resident stent cells, thereby supporting the myocardial healing mechanisms after injury. Administration of autologous hematopoietic stem cells or mobilization of endogenous stem cells has been shown to be safe after myocardial infarction or cardiomyopathies, whereas skeletal myoblasts are considered to be hazardous due to the occurrence of life-threatening arrhythmias. This review focuses on the use of adult human stem cells for treating myocardial infarction and cardiomyopathy, and discusses recent preliminary efficacy data, which suggest that 'regenerator cells' might have the potential to improve myocardial perfusion and contractile performance in patients suffering from myocardial infarction, severe ischemic heart disease and chronic heart failure. PMID- 17078383 TI - Gene therapy for chronic granulomatous disease. AB - Chronic granuloniatous disease (CGD) is a rare inherited imnunodeficiency characterized by recurrent, often life threatening bacterial and fungal infections due to a functional defect in the microbial-killing activity of phagocytic neutrophils. If regular care and conventional therapy fail, tile disease can be cured by bone marrow transplantation. This treatment is, however, only available to patients with human leukocyte antigen-identical sibling or matched unrelated donors. One therapeutic option for patients lacking suitable donors is the genetic modification of autologous hematopoietic stem cells. This review discusses the developments that have led to the realization of a successful gene therapy protocol for the correction of CGD. PMID- 17078384 TI - New serotypes of adenoviral vectors. AB - Since it was first demonstrated that recombinant adenovirus (Ad) vectors could be generated by inserting foreign genes into Ad genomes, they have been extensively used as eukaryotic expression vectors and therapeutic gene delivery vehicles. Originally, Ad vectors were based on the human Ad5 serotype and while most currently used Ad vectors are still based on Ad5, recent research has highlighted a series of problems that limit the efficacy and safety of these vectors. To circumvent the problems associated with Ad5-based vectors, new vectors have utilized various Ads of both human and nonhuman origin. In this review we introduce the recent advances made in the development of non-Ad5-based vectors. PMID- 17078385 TI - Gene therapy of surfactant protein B deficiency. AB - Surfactant protein B (SP-B) is encoded by a single gene, and the mature protein is expressed by alveolar type II epithelial (ATII) cells of the lungs. Studies in transgenic mice and its hereditary deficiency in humans have established its indispensable role in postnatal survival. An established gene therapy regime for treating SP-B deficiency could overcome the limitations of surfactant replacement therapies or lung transplantation. Among the various viral and non-viral gene delivery tools available, only adenoviral vectors have been tested for delivering SP-B cDNA to the lungs of animal models. This review discusses the various vectors that are available for delivering therapeutic genes into ATII cells. PMID- 17078386 TI - Gene therapy for cystic fibrosis lung disease: current status and future perspectives. AB - Cystic fibrosis (CF) is an autosomal recessive genetic disease affecting > 70,000 individuals worldwide. Despite improvements in current therapies, most patients do not survive beyond their early 30s. After cloning of the cystic fibrosis transmembrane regulator (CFTR) gene, there was considerable clinical interest in the possible therapeutic delivery of CFTR genes directly to the lung. Several clinical studies have since demonstrated proof-of-principle for correction of the underlying chloride defect in CF patients using viral and non-viral vectors. Inefficient gene transfer and host-antigen-specific immune responses caused by replication-deficient viral vectors have elevated non-viral approaches to becoming the field's most promising therapeutic contenders. Among these non-viral gene therapy vectors are cationic liposome/plasmid DNA complexes and compacted DNA nanoparticles carrying the CFTR gene, which have shown promise for the treatment of CF in phase I clinical trials. However, the levels of CFTR expression achieved in the respiratory epithelium were too low and only of limited duration. Improved strategies for efficient and prolonged expression of the transgene are therefore necessary. This review outlines the current repertoire of available gene vectors and discusses novel strategies to enhance the efficiency and selectivity of gene transfer for gene delivery into the lung. PMID- 17078387 TI - Progress towards the clinical application of helper-dependent adenoviral vectors for liver and lung gene therapy. AB - This review focuses on recent developments in helper-dependent adenoviral technology and preclinical studies for helper-dependent adenovirus-mediated liver and lung-directed gene therapy. Studies highlighting the tremendous potential of these vectors are reviewed, together with some important obstacles that will need to be addressed before clinical utility. PMID- 17078388 TI - Drug evaluation: tefibazumab--a monoclonal antibody against staphylococcal infection. AB - Inhibitex Inc is investigating tefibazumab, a humanized monoclonal antibody specific for the fibrin-binding surface epitope clumping factor A protein (expressed on the surface of most Staphylococcus aureus strains), for the potential intravenous prevention and/or treatment of S aureus infections. In June 2006, Inhibitex was seeking to outlicense certain development rights to the drug. Pending the outcome of partnering discussions, Inhibitex suspended the initiation of any additional clinical trials of tefibazumab. Preclinical in vivo studies were ongoing at that time. PMID- 17078389 TI - Drug evaluation: ISIS-301012, an antisense oligonucleotide for the treatment of hypercholesterolemia. AB - ISIS-301012 is an antisense oligonucleotide inhibitor of apolipoprotein B-100, which is being developed by Isis Pharmaceuticals Inc for the potential treatment of hypercholesterolemia. A subcutaneous injectable formulation is currently undergoing phase 11 clinical trials, while phase I trials are underway with an oral formulation of the drug. PMID- 17078390 TI - [The history of the General Hospital of Madrid (Hospital Provincial), house of a Centennial Urology Department]. AB - OBJECTIVES: To show the beginning of the General Hospital of Madrid, its localization, economical problems and medical protagonists, mainly the first medical director: Dr. Perez de Herrera. Also, to show its scientific and health care contributions to the people of Madrid and other cities, coming in the search for training or health-care. METHODS: We reviewed writings and books from well known doctors, experts on history of medicine, and documents about the old and new Madrid and its monuments. RESULTS AND CONCLUSIONS: The history of this hospital, medical school and temple, is engaged with the history of Madrid's provincial Charity. Economical difficulties were great to subside. At the end of the 19th century one of the eldest urology departments in Spain was consolidated. PMID- 17078391 TI - [Contribution to the anatomical-morphological study of the pelvic floor in the asymptomatic female: the use of MRI imaging]. AB - OBJECTIVES: To confirm the results of previous studies demonstrating the morphology of the levator ani muscle in the living subject is different to that described in classic anatomical works; to evaluate the anatomical-morphological differences of the pelvic floor between nulliparous and multiparous women in order to analyze the influence of pregnancy and delivery. METHODS: Comparative study of the morphological variations of the iliococcigeous fascicle of the levator ani muscle between two groups of females using T2 MRI.: the first group included 11 nulliparous women and the second group 9 multiparous women. The curvature radium of the fascicles was calculated in the frontal projections. The differences in height were also calculated. The anterior limit of the iliococcygeal muscle dome was determined in the sagittal plane. The software Image Tool 3.0 was employed for the measurements. The Kolmogorov-Smirnov test was employed to analyze the distribution of the study populations; mean value comparisons between groups were performed by the Student's t test. Finally, the various morphological measurements were compared in relation to various parameters: parity, side, body mass index (BMI), sports practice, menstrual cycle phase, and presence or absence of episiotomy. Data obtained were analyzed using the Fisher's exact test, with a statistical significance of p < 0.05. RESULTS: Morphological measurements: We observed that the concavity of the iliococcygeal fascicle dome is larger in nulliparous women (p = 0.03 for the right side and p = 0.04 for the left). Moreover, these women have the domes significantly more anterior (p < 0.001 for both sides). Comparisons between other variables: an association between nulliparous status and the presence of a higher and more anterior left dome, and multiparous status and a higher and more anterior right dome were (p = 0.02). CONCLUSIONS: The hypothesis of the pelvic floor morphology being a double dome with inferior-posterior concavity in the living asymptomatic female is confirmed. We also demonstrate the existence of anatomical morphological differences in the iliococcygeal muscle of the levator ani between nulliparous and multiparous females, which seems to confirm a relationship with pregnancy and/or delivery. PMID- 17078392 TI - [Inverted papilloma of the bladder]. AB - OBJECTIVES: To report a series of patients with bladder inverted papilloma, pointing out the clinical features, presentations, treatment options and outcomes. We also perform a bibliography review oriented to the diagnostic criteria, treatment options and prognosis of the inverted papilloma. METHODS: We reviewed the medical charts of all patients with the diagnosis of bladder inverted papilloma treated by our group between July 1996 and June 2003. RESULTS: 16 cases of inverted papilloma of the bladder were diagnosed in ten males and six females, with a mean age of 53.1 yr. (range 31-72). The most frequent reason for consultation was voiding urgency (43.7%), followed by hematuria (37.5%). In 14/16 patients the lesion was localized in the bladder trigone. Transurethral resection of the lesion was performed in all patients. Two cases (12.5%) presented associated transitional cell carcinoma. CONCLUSIONS: The inverted papilloma is a rare pathologic entity Despite the absence of agreement about its aggressiveness or innocuousness we consider its potential malignancy should be taken into consideration after the evidence of recurrences and its association with transitional cell carcinoma. PMID- 17078393 TI - [Immunosuppression and surgical complications after renal transplantation]. AB - OBJECTIVES: The new immunosuppressive regimens in kidney transplantation have diminished the rate of acute rejection and improved graft survival. However, the use of new agents results in the development of surgical complications. The authors analyze the incidence of such complications accordingly to the type of drug. METHODS: This study included 350 kidney transplantations performed between January 1997 and December 2004. The average age was 54 years. The incidence of diabetes mellitus was 8.5% and the rate of obese recipients (BMI >30 kg/m2) was 15.4%. The average follow-up rate was 44 +/- 5.6 months (5-96). A surgical complication was defined as any complication directly related to the surgical transplant that occurs along the first year after transplantation. RESULTS: The incidence of surgical complications was 34.8% (122/350). The rates of perigraft collections and bleeding posttransplant were significantly higher in the CsA group than in the Tacro one: 12% vs. 3.8% (p = 0.005) and 11.5% vs. 3% (p = 0.002). The Sirolimus and Everolimus-based immunosuppresive regimens led to a higher incidence of lymphocele (16% vs. 3.7%) (p = 0.012). There were no significant difference in the incidence of surgical complications in recipients immunosuppressed with and without MMF, and in diabetic vs. no diabetic patients. Surgical complications were higher in obese patients (66.5% vs. 33.5%) (p = 0.002). CONCLUSIONS: Recipients receiving Sirolimus/Everolimus demonstrated a significantly higher lymphocele rate. The CsA regimens were associated with bleeding and post transplant pararenal fluid collections. The introduction of MMF did not result in a significant increase in posttransplant surgical complications. The obesity was a risk factor associated with those complications. PMID- 17078394 TI - [Urodynamic factors associated to vesicoureteral reflux in adults]. AB - OBJECTIVES: Lower urinary tract dysfunction may affect the upper urinary tract with development of vesicoureteral reflux. The objective of our study is to evaluate what lower urinary tract dysfunctions are associated with vesicoureteral reflux in adults. METHODS: We performed a cross-sectional study in a sample of 236 patients (149 males and 87 females) with a mean age of 45.5 yr. (typical deviation 19.0 yr.) submitted for videourodynamic study for lower urinary tract functional symptoms. History and neurourologic physical examination, filling cystometry associated with videocystography, pressure/flow study associated with voiding videocystography and selective electromyography of the periurethral sphincter were done in all patients. The chi-square test was used for statistical analysis. RESULTS: The presence of vesicoureteral reflux was observed in 19 patients (8.1% of the sample). Fifteen cases (79%) showed reflux during the filling phase, 3 cases (16%) during the voiding phase, and in 1 (5%) in both. The existence of a vesicourethral neurogenic dysfunction was confirmed in 8 cases (42%); there was no neurogenic dysfunction in 11 cases (58%). The vesicoureteral reflux was classified as primary passive (without any urodynamic anomaly) in one case (5%), passive with diminished compliance in 4 cases (21%), passive evolutive in 3 cases (16%), active involuntary in 7 cases (38%), active associated with organic obstruction of the lower urinary tract in one case (5%), active associated with increased contractile potency in one case (5%), and secondary to abdominal press in 2 cases (10%). The only urodynamic anomalies associated with a significant increase of the prevalence of vesicoureteral reflux were: a filling pressure at maximum capacity greater than 12.5 H2O cm (prevalence of reflux 7 times) and voiding with abdominal press which increased the prevalence of reflux 2.8 times. CONCLUSIONS: Most patients with functional symptoms of the lower urinary tract and associated vesicoureteral reflux present lower urinary tract dysfunctions that may justify their reflux. Nevertheless, a statistically significant increase of the prevalence of vesicoureteral reflux was only found in patients with filling pressures at maximum capacity greater than 12.5 cm H2O and those voiding with the help of abdominal press. PMID- 17078395 TI - [Periurethral injections of dextranomer/hyaluronic acid copolymer in the treatment of female stress urinary incontinence: description of the technique and bibliographic review]. AB - OBJECTIVES: The system enables a non endoscopic, outpatient treatment of the female stress urinary incontinence. The article intends to familiarize the readers with the procedure. METHODS: We describe the technique, graphically, with all steps, and perform a bibliographic review. RESULTS: In accordance to the main clinical studies reviewed, the periurethral injection of dextranomer/ hyaluronic acid copolymer has an efficacy of near 75%, with a simple, reproducible technique with rare adverse events. CONCLUSIONS: The treatment with this type of periurethral injections may be valid as a minimally invasive treatment of the female stress urinary incontinence, because of its efficacy, tolerability and reproducibility. PMID- 17078396 TI - [Single trocar laparoscopic nephrectomy]. AB - OBJECTIVES: The usual laparoscopic hand assisted nephrectomy procedure requires either two or three trocars and a small incision to extract the organ. Our experience reveals that surgical invasion is significantly less when we use only one trocar and a retroperitoneal approach. METHODS: We start with a small low oblique incision close to the pubis and insert a trocar over the iliac crest. When the kidney is free we grasp the undissected vascular pedicle between thumb and middle finger and without moving the inserted hand, we remove the telescope and introduce the stapler. The index finger is used blind to check that the stapler is correctly positioned before triggering. Finally, we divide the ureter in the same way We have carried out 74 nephrectomies with this procedure. RESULTS: The mean operative time was 67.12 minutes. A retroperitoneal haematoma occurred in one patient and was drained by the same approach without further incident. One patient had an intestinal occlusion that in the reoperation was found to be caused by a loop of intestine trapped in a peritoneal tear: the postoperative was satisfactory. We lost one patient: an 89-year-old lady with a pyonephrosis and a retroperitoneal abscess. The operation was uneventful, but the patient died after five days; possibly because of septic shock. The other patients had no significant complications. CONCLUSIONS: The use of only one trocar markedly reduced operative invasion. The blind division en-bloc of the pedicle saves valuable time and does not offer any additional risk. The possibility of an arterio-venous fistula is remote because the stapler leaves the artery close to the vein, but separated from it. Cost is minimal because the surgeon only needs one trocar and a stapler. Consequently, we believe that this procedure with only one trocar could also be safely adopted for living donor nephrectomy PMID- 17078397 TI - [Idiopathic granulomatous orchitis: pathologic study of one case]. AB - OBJECTIVE: To report one case of idiopathic granulomatous orchitis, an extremely rare disease, in a 76-year-old patient. METHODS/RESULTS: The pathology department received a testicle with the clinical/radiological diagnosis of testicular tumor. The pathologic study showed absence of neoplasias and presence of morphological findings compatible with idiopathic granulomatous orchitis. CONCLUSIONS: The idiopathic granulomatous orchitis is an entity of unknown etiology, clinically or ultrasonographically not distinguishable from testicular neoplasias, the diagnosis of which is made after orchiectomy. PMID- 17078398 TI - [Penile leiomyosarcoma: case report and bibliographic review]. AB - OBJECTIVE: Primary penile leiomyosarcoma is a rare entity. Since 1930 only 30 cases have been reported in the bibliography. We wanted to add a new case to the international literature, in addition to a review of all available publications on the topic from 1957. A 54-year-old patient presented with a lobulated 8x4x3 cm lesion in the balanopreputial groove over two years; he was treated initially with partial penectomy and subsequently with total penectomy We analyze the evolution, progression and adjuvant treatments of this rare pathology. METHODS/RESULTS: We report one case with its clinical presentation, diagnostic tests performed for staging, treatment and follow-up. CONCLUSIONS: The first case of penile leiomyosarcoma was described by Levi in 1930. In 1957 Ashley and Edwards reported the first case in the British literature and in 1963 Pack reported the first in the American literature. MacKenzie et al. were the first to recognize two types of leiomyosarcoma: superficial and deep. The first develop from smooth muscle cells from the superficial dermal layers of the glans penis or distal third of the penis, they are generally asymptomatic and less malignant, rarely invading deeper structures and without involvement of the urethra on physical exam. The best prognostic predictors are type of tumor at presentation (superficial vs. deep) and treatment choice, being total penectomy more effective for failures of local resections than for primary treatment of deep tumors. PMID- 17078399 TI - [Penile fracture with associated urethral rupture]. AB - OBJECTIVE: We report two cases of penile fracture; one case with associated rupture of the corpus spongiosum and complete urethral section, and the other presenting a second cavernous fracture three years after a first episode. METHODS: The first patient, 32-years-old, suffered the fracture of the right corpus cavernosum during intercourse three years after a first fracture requiring surgical repair. This time he also underwent surgery immediately to find and repair the lesion, with good functional outcome. The second patient, 29-years old, suffered a transverse fracture at the bases of both corpora cavernosa associated with complete section of the urethra. He underwent emergency surgery to repair all lesions with good esthetic and functional results. RESULTS: Traumatic lesions of the penis with fracture of the corpora cavernosa are rare and the association with urethral rupture is exceptional, as well as it is the possibility of second fracture years ofter a first episode. CONCLUSIONS: Penile fracture is a rare reason for consultation in the emergency setting, which should be repaired immediately with evacuation of the hematoma, closure of the albuginea's lesion and repair of the urethral lesions when they appear. Results are better, avoiding mid and long-term complications. PMID- 17078400 TI - [Metastases from a rectal adenocarcinoma to the prepuce]. AB - OBJECTIVE: We report one case of metastatic dissemination of a rectal adenocarcinoma to the prepuce. METHODS: 61-year-old patient with the diagnosis of rectal adenocarcinoma treated 18 months before by surgery and chemotherapy. He presents with a painful enlargement of the penis associated with outgrowing erythematous lesions in the skin of the prepuce that bled on touch and did not allow the vision of the meatus and the glans penis. Physical examination showed the presence of hypogostric, penile and scrotal lymphedema. Circumcision was performed. RESULTS: The pathologic study reported a moderately differentiated intestinal type adenocarcinoma with high mitotic index infiltrating the squamous cell flat epithelium of the prepuce. CONCLUSIONS: Although extremely rare, tumor implants in the prepuce secondary to extra urologic tumors are exceptional. Surgical excision confirms the origin and may avoid bleeding and discomfort, and also may help with catheterization, which is many times necessary in the final stages. PMID- 17078401 TI - [Renal hydatidosis]. AB - OBJECTIVE: Our purpose is to report a case of renal hydatidosis and its treatment. METHODS: The characteristics of the case are presented and discussed. RESULTS AND CONCLUSIONS: The hydotic disease is a parasitic infestation caused by the larvae from echinococcosis granulosus. The kidney ranks third among all visceral localitations, compromising only 2% to 4% of all cases. Most of patients are asymptomatic for years and the correct preoperative diagnosis is difficult. PMID- 17078402 TI - [Prostate cancer and bilateral synchronic renal tumors. Case report]. AB - OBJECTIVE: To report one case of synchronic prostate cancer and bilateral renal tumors, radiologically suggestive of renal cell carcinoma. METHODS: We present the case of a 63-year-old diabetic male, heavy smoker, who presents at the outpatient office complaining of right upper quadrant abdominal pain and mild obstructive voiding symptoms. Abdominal examination was negative and digital rectal examination revealed a rock hard irregular prostate. RESULTS: The prostatic biopsy showed a moderately differentiated prostate adenocarcinoma. Abdominal ultrasound showed the existence of a 6 cm heterogeneous complex tumor at the lower pole and middle portion of the right kidney, and another at the upper pole-middle left kidney. CT scan confirmed the tumors, which enhanced with contrast. Chest x-ray and bone scan were negative for metastases. Two fine-needle aspiration biopsies of the right kidney tumor were unsuccessful and patient refused to undergo further diagnostic procedures. Treatment for the prostate cancer with LHRH analogues and antiandrogenic drugs was started. The three-month follow-up CT scan showed no variation. CONCLUSIONS: Despite the absence of a histological diagnosis due to refusal of the patient, CT scan images are typical of bilateral renal cell carcinoma, supporting the idea that in the presence of a genitourinary neoplasia we should suspect and rule out a synchronic second tumor of the same apparatus. PMID- 17078403 TI - [Metastasis of colon adenocarcinoma to the epididymis and spermatic cord]. AB - OBJECTIVE: To report the case of an epididymal and spermatic cord metastasis of a colon adenocarcinoma. METHODS/RESULTS: We present the case of a 67 years old man, who presents several months after subtotal left colectomy for a left colon adenocarcinoma (pT3 NoMo), with a symptomatic right palpable testicular tumour. Right orchiectomy was done, demonstrating colon adenocarcinoma metastasis. CONCLUSIONS: Epididymal and spermatic cord metastases are very infrequent and they usually are found incidentally after prostate cancer orchyectomy. PMID- 17078404 TI - [Acute pyelonephritis in ectopic crossed and fused kidney]. PMID- 17078405 TI - [Seminal vesicle cyst and ipsilateral renal agenesis]. PMID- 17078407 TI - A new form of presentation of renal actinomycosis: renal tumor with retroperitoneal bleeding. AB - OBJECTIVE: The aim of this case report is to add to the literature a new case of renal actinomycosis, but with a form of presentation that has never been reported: renal tumor with retroperitoneal bleeding. METHODS/RESULTS: We present the case of a 27-year-old woman, with a 5-month history of general syndrome and right flank pain. Radiological findings showed a right renal tumor with suspicious of retroperitoneal bleeding. Right radical nephrectomy was performed and the pathological examination of the specimen found actinomyces colonies. The patient received 8 weeks of Penicillin after surgery and had none sequelae. easily diagnosed because of non-specific clinical and radiological findings. Early diagnosis of renal actinomycosis is important to avoid surgery since actinomyces respond well to high doses of Penicillin. PMID- 17078408 TI - The intersection of patient safety and nursing research. AB - The individual and collective discussions of the patient safety issue in the United States have mounted from a low roar to a deafening din in the past 10 years. In this chapter the authors (1) discuss the context of patient safety over the past decade and the federal response to the problem, (2) briefly present Reason's theory of human error, which frames much of the safety research, and (3) provide a glossary of terms. PMID- 17078409 TI - Medication-related errors: a literature review of incidence and antecedents. AB - Patient safety has become a major concern for both society and policymakers. Since nurses are intimately involved in the delivery of medications and are ultimately responsible during the medication administration phase, it is important for nursing to understand factors contributing to medication administration errors. The purpose of this chapter is to identify the incidence of these errors and the associated factors in an attempt to better understand the problem and lessen future error occurrence. Literature review revealed both active failures and latent conditions established in Reason's theory remain prevalent in current literature where active failures often display themselves in the form of incorrect drug calculations, lack of individual knowledge, and failure to follow established protocol. Latent conditions are evidenced as time pressures, fatigue, understaffing, inexperience, design deficiencies, and inadequate equipment and may lie dormant within a system until combined with active failures to create opportunity for error. Although medication error research has shifted in emphasis toward identification of system problems inherent in error occurrence, no one force emerges as a clear antecedent, reinforcing the need for further research and replication of existing studies with emphasis placed on more dependable reporting measures through which nurses are not threatened by reprisal. PMID- 17078410 TI - Fall and injury prevention. AB - Falls and related injuries are increasingly being recognized as a nursing sensitive quality indicator, and they continue to be an unsolved patient safety problem in inpatient and outpatient care areas as well as in the community at large. The purpose of this review is to summarize the current research related to fall and injury prevention. The chapter is organized presenting research in (1) the community and (2) acute and long-term care settings. For each setting, the research that addresses risk factors, risk assessment instruments, and fall and injury prevention efforts are reviewed. There is a large body of research that investigates fall and injury prevention across the care continuum. In the community setting, targeted risk evaluation in the emergency department and management of vitamin D deficiency appear to be promising preventive methods. However, further research needs to explore staffing ratios, automated methods of assessing and communicating fall risk, improved methods and timing of risk evaluation and methods by which existing and new evidence might be translated into practice. PMID- 17078411 TI - Hospital-acquired infections as patient safety indicators. AB - Transmission of infection in the hospital has been identified as a patient safety problem adversely affecting patients, visitors, and health care workers. Prevention of infection should not be limited to the hospital epidemiology staff but also must involve the entire multidisciplinary team, including nurses. This chapter reviews the literature related to patient safety of nursing-authored studies of infection control in the hospital. The review indicated that there were key areas of research interest including drug resistance; hand hygiene products, procedures, and surveillance; preoperative skin preparations; health care worker transmission of infection; common procedures associated with an increased risk of transmission; and organizational issues. PMID- 17078412 TI - Patient safety in hospital acute care units. AB - The most visible threats to patient safety associated with nursing care occur on hospital inpatient units. Patient safety research is a new phenomenon, but it builds on the knowledge provided by quality-of-care research done previously. The purpose of this chapter is to describe the current state of the science in the area of nurse staffing and patient safety. The results of research studies published since the last round of reviews (1996-2005) are described by level of analysis, measures of nurse staffing and patient outcomes. Although research linking nurse staffing to the quality of patient care has increased markedly since 1996, the results of recent research projects do not yet provide a thorough and consistent foundation for producing solutions to the crisis in hospital nursing care. The inconsistencies are largely due to differing units of analysis (hospital, patient, care unit), variability in measures of nurse staffing, the variety of quality indicators chosen, the difficulty finding accurate measures of these indicators, and the difficulty creating risk-adjustment strategies for the indicators most sensitive to nursing care. Nursing administration and policy most urgently need research conducted with standardized data collected at the patient care unit level. PMID- 17078414 TI - Nurse home visit programs for the elderly. AB - Nurses are the largest professional provider of health care services in the home setting. However, nurse home visit programs are diverse. The purpose of this review was to examine the many factors that influence the effectiveness of nurse home visit programs for older adults. Donebedian's Quality Assessment Model was used to organize the review using the components of structure, process, and outcome. A total of 60 home visit studies were identified that met the following criteria: (1) nurses were a major or only provider of the intervention, (2) the intervention was delivered by home visits, and (3) the study included a comparison group. This review demonstrates the complexity of variables that determine the effectiveness of home visit interventions. Many studies demonstrated lower overall health care costs for the intervention group with either improved or at least no change in clinical outcomes. PMID- 17078413 TI - Medication safety within the perioperative environment. AB - With the widespread patient safety movement comes an increased public awareness of the risks inherent within the health care setting. More specifically, the highly publicized medication error cases that hit the media demonstrate the effect mediation errors have on patient safety within the perioperative environment. This awareness, however, has triggered limited research across the continuum of care within this complex environment. A current review of the state of the science related to medication safety within this setting reveals research primarily focused on the anesthesia domain of practice. Although application to the perioperative environment can be extrapolated from this research, there is a notable lack of nursing-initiated research that focuses on improved systems or processes related to medication safety within the perioperative continuum of care. This knowledge gap in the literature presents an excellent opportunity for nursing to grow a research program to improve medication safety within the perioperative environment in support of evidence-based practice. PMID- 17078415 TI - Nursing home safety: a review of the literature. AB - The number of older persons in the United States is rapidly growing and, based on this growth projection, the number of consumers needing nursing home (NH) care will likely triple in the next 10 years. Although NHs have been bombarded and scrutinized about the care that they provide, the concept of safety (specifically, error prevention) remains at the margin of most quality improvement efforts. The purpose of this review is to explore what has recently been written (2000-2005) about the evolution of the NH as an organization focused on safety and the most critical clinical processes that must be closely monitored for a safe NH environment to occur. After a thorough review of both organizational and clinical NH literature, 30 organizational studies and 39 clinically based studies were reviewed. The review revealed that, organizationally, teamwork, communication, and leadership all were critical in resident and staff outcomes and clinically, assessment was an important missing process at critical points in the residents' care for prevention and timely treatment of potentially dangerous conditions. The value of the registered nurse (RN) in this setting was clear in the many assessment issues noted and the lack of RN guidance for adherence to recognized practice guidelines. To explicate the role of the RN, first, better outcome measures must be developed that are nurse sensitive. A second clear agenda for NH research is the explication of the role of leadership, particularly nursing leadership, to create an environment where open and accurate communication can be accomplished among all of the diverse NH roles. This will help all members of the team to identify care improvement opportunities. Finally, a new frontier for the NH setting is the use of technology and the need to harness the information that has set in the NH system for years. Information mastery for staff and leadership is a necessary aspect of the organization that must be developed to provide sound information for strategic and focused change to occur. PMID- 17078416 TI - Informatics for patient safety: a nursing research perspective. AB - In Crossing the Quality Chasm, the Institute of Medicine (IOM) Committee on Quality of Health Care in America identified the critical role of information technology in designing a health system that produces care that is "safe, effective, patient-centered, timely, efficient, and equitable" (Committee on Quality of Health Care in America, 2001, p. 164). A subsequent IOM report contends that improved information systems are essential to a new health care delivery system that "both prevents errors and learns from them when they occur" (Committee on Data Standards for Patient Safety, 2004, p. 1). This review specifically highlights the role of informatics processes and information technology in promoting patient safety and summarizes relevant nursing research. First, the components of an informatics infrastructure for patient safety are described within the context of the national framework for delivering consumer centric and information-rich health care and using the National Health Information Infrastructure (NHII) (Thompson & Brailer, 2004). Second, relevant nursing research is summarized; this includes research studies that contributed to the development of selected infrastructure components as well as studies specifically focused on patient safety. Third, knowledge gaps and opportunities for nursing research are identified for each main topic. The health information technologies deployed as part of the national framework must support nursing practice in a manner that enables prevention of medical errors and promotion of patient safety and contributes to the development of practice-based nursing knowledge as well as best practices for patient safety. The seminal work that has been completed to date is necessary, but not sufficient, to achieve this objective. PMID- 17078417 TI - Organizational climate and culture factors. AB - Nurses and others have expressed a great deal of interest in the potential for incorporating notions about organizational culture and climate in research and practice aiming to improve health care safety. In this review, definitions and measures of these terms are explored, the state of the research literature connecting culture and climate with safety is reviewed, and directions for future research and leadership practice are outlined. PMID- 17078418 TI - Methodologies used in nursing research designed to improve patient safety. AB - Nursing research studies of patient safety for 2002-2005 were reviewed to determine methods used and methodological challenges within this field of research. Methods used in traditional clinical research and in health services research were often combined or adapted in innovative research designs to advance knowledge regarding nursing care and patient safety outcomes. This relatively new focus of complex research posed methodological challenges in areas such as measurement and the availability and analysis of data. The most frequent methods used included survey research, analysis of secondary data, and observational studies. This review points to the need to increase the incorporation of complex methodological training, including health services research, the analysis of secondary data and complex survey design in our doctoral programs, and to provide opportunities for researchers to gain further methodological training. Increased use of multi-site and multi-level studies is also needed. PMID- 17078419 TI - Nursing research in Ireland. AB - This review presents an analysis of research published by Irish nurses during the period 1990-2005. The analysis is the first effort made to identify the main characteristics of Irish nursing research. Overall, 213 published studies were identified for consideration, from which, 152 were included in the review. The studies were published in 60 journals, 4 books, and 8 research reports. Journal articles selected from 6 journals accounted for 52%. Inclusion criteria were quality of research design, sampling (including sample size), data analysis, scientific merit, and authorship. Each article was analyzed based on this schema. The major areas of research identified included clinical practice (56%) (e.g., medical surgical, mental health, intellectual disability, and maternal and child), nursing management and professional issues (19%), and nursing education (25%). PMID- 17078420 TI - [Ethical considerations of psychiatric treatment in chronic post traumatic stress following multiple casualty attacks]. AB - Israeli society has been repeatedly exposed to traumatic stress following terrorist attacks with multiple casualties. Individuals who are subjected to these incidents experience various mental reactions, including traumatism, fear, loss and bereavement. Unfortunately, evidence based medical knowledge is lacking regarding natural methods for dealing with incidents that may precipitate traumatic stress, ways of identifying people who are more likely to develop post traumatic symptoms, or suggestions on early interventions that may avert the onset of these symptoms. Ethical problems may arise, especially in situations involving mass traumatic stress, when there are insufficient therapeutic resources. In such cases, the issues of whether to intervene, who to treat, and how, will often be influenced by conflicting personal and social interests. Another concern involves the principle of autonomy - the individual's right to decide whether he/she wants treatment. Should "latent" victims be identified in the community and referred to therapy? Diverse ethical issues emerge in situations of traumatic stress, and may potentially evoke dilemmas among health policy makers and among the therapists who work in the field. This type of discussion may enhance the understanding, help formulate key principles and assist in making reasonable and appropriate decisions when dealing with such difficult and complex situations. PMID- 17078421 TI - [The effects of violence and terrorism on chronic physical illness]. AB - Over the past few years more than a thousand Israeli civilians were murdered and thousands were wounded in recurrent terrorist attacks. Many others had been exposed to the terrorists' attacks, either directly or indirectly. However, the impact of the terror on physical illness had not yet been explored. Stress may cause diseases like hypertension or ischemic heart diseases and may exaggerate asthma, inflammatory bowel diseases and others; this is in addition to posttraumatic disorders, depression and anxiety. This article reviews the influence of stress, and especially conflicts, on chronic physical illness and aims to encourage further research on this subject in Israel. PMID- 17078422 TI - [The Objective Structured Clinical Examination and its potential applications as an assessment and feedback tool for IDF primary care physicians--a pilot study]. AB - BACKGROUND: In recent years, the Israel Defense Forces IDF developed special training programs for junior military physicians at the beginning of their service and CME courses for senior military physicians. The Objective Structured Clinical Examination (OSCE) is a well established assessment tool in medical education that can be of vast use in military training programs for primary care physicians. AIM: To evaluate the OSCE as an assessment tool of military primary care physicians. METHODS: Forty-one military primary care physicians completed a 10-station OSCE. The clinical scenario choices were based on the most prevalent problems in the annual statistics of the Israeli Medical Corps between the years 1988-1998. Each station consisted of two parts: a doctor-SP (standardized patient) encounter and a written exercise. Both parts were evaluated by checklists designed and validated by a committee of OSCE experts and senior military physicians. RESULTS: The overall reliability of the exam was alpha = .88. The examinees highly valued the relevance of the scenarios to their every day practice (4.38 out of 5.0) and the reliability of the standardized patients (4.0 out of 5.0). A total of 92% request feedback on their performance. CONCLUSIONS: This pilot study found the OSCE to be a feasible, valid and reliable tool that can serve to assess the capability of physicians to deal with major issues in military primary health care. The OSCE can also play a major role in structured feedback provided to these physicians. PMID- 17078423 TI - [A Hebrew version of the FLACC scale: measurement of pain in non-verbal children]. AB - BACKGROUND: The Israeli Ministry of Health requires that every patient have their pain routinely and systematically measured when there are treated in any of the country's medical institutions. Measurement guides treatment and enables follow up of pain over time. Self-assessment of pain is the gold standard. Measurement is standardized by using scales representing intensity from "no pain" to "unbearable pain". Three-year-olds can assess their own pain, but younger children, or those who are non-verbal due to a medical procedure, cannot. Scales for this population rely on behavioral and physiological parameters, with assessment conducted by caretakers. Of the scales reviewed by the authors, the "FLACC" was chosen as appropriate for routine use due to its brevity and simplicity. AIMS: Translation and establishment of reliability and validity of the Hebrew version of the FLACC. METHODS: Subjects included 53 children aged 2 months to 8 years, who could not communicate verbally. Observations were conducted in the post-anesthesia care unit and the Intensive Care Unit. The FLACC was translated by the translation-back-translation technique. Inter-rater reliability was tested by two independent observers, and validity was assessed before and after provision of intravenous morphine or ketorolac. RESULTS: Inter rater reliability was high for the total FLACC score (r = 0.94, p < 0.001), as well as for the separate items (kappa 0.5-0.85). The FLACC was considered valid, as the change in scores paralleled the known pharmacological effect of the medications. CONCLUSIONS: The Hebrew version of the FLACC was found reliable and valid for caretakers to use with children who cannot communicate verbally. PMID- 17078424 TI - [Medical rehabilitation of patients with osteogenesis imperfecta (OI) and additional newly acquired disabilities]. AB - Adults with Osteogenesis Imperfecta (OI) are rarely referred to facilities of general rehabilitation medicine. Recently, two OI patients were admitted to our department because of additional neurological disabilities: stroke and spinal cord injury. In these circumstances, the hospitalization course has different characteristics. PMID- 17078425 TI - [Transcatheter treatment of combined congenital heart anomalies in an adult cyanotic patient]. AB - Contemporary catheterization techniques allow effective percutaneous transcatheter treatment of congenital heart defects as an attractive alternative to open heart surgery. This case report describes transcatheter treatment of congenital pulmonary valve stenosis and secundum atrial septal defect which were diagnosed in a 31 year old woman who presented with cyanosis. This illustrates difficulties in the diagnosis of congenital heart disease in adult patients and demonstrates that transcatheter techniques in the treatment of these congenital anomalies are effective. PMID- 17078426 TI - [Two cases of transient global amnesia (TGA) following sildenafil use]. AB - Only two cases connecting Sildenafil reception and acute memory impairment have been published. Two similar cases were observed in our clinic last year. Sildenafil is a potent inhibitor of cyclic guanosine monophosphate in the corpus cavernosum and therefore, increases the penile response to sexual stimulation and is used for erectile dysfunction. The most severe and life-threatening complications of Sildenafil are associated with combined administration with nitrates. The incidence of nonfatal myocardial infarction, stroke and death did not significantly differ between Sildenafil-treated and placebo-treated patients; therefore, Sildenafil does not appear contraindicated in subjects with ischemic heart disease (IHD). Scanty data are available regarding Sildenafil and cerebrovascular disease and there are only a few case reports regarding transient global amnesia (TGA) after Sildenafil use. Two cases of TGA are described immediately following the use of one dose of Sildenafil. The etiology of TGA is not yet completely understood but one of the hypothesizes suggests that the pathophysiology of this condition is related to intracranial vasomotor changes, especially due to venous congestion and venous ischemia of bilateral hippocampal structures. It is also well known that Sildenafil stimulates the relaxation of smooth muscle and causes vasomotor changes. Based on this report, as well as previous reports, it is suggested that a single dose of Sildenafil may stimulate TGA. PMID- 17078427 TI - [Limited geriatric assessment of the elderly in Kibbutzim of the Upper Galilee]. AB - BACKGROUND: Since comprehensive geriatric assessment (CGA) is very time and money consuming, its implementation is limited in Israel. OBJECTIVE: Implementation of the ambulatory limited geriatric assessment (LGA). TARGET POPULATION: Persons over 65 years of age living in Kibbutzim in the Upper Galilee. METHODS: As in the case of CGA, the emphasis in LGA is mainly placed on the functional status of the elderly. In order to identify geriatric syndromes, we used routine screening methods. According to the agreement with Clalit Health Services, a geriatrician is allowed to spend one hour per person. The elderly to be checked were selected by the medical staff. At the time of LGA, medical staff and patients' relatives supplied all information needed, including this data in computerized ambulatory cards. In such a way, a considerable amount of medical, functional, epidemiological, and psychosocial information was retrieved thus enabling a geriatrician to elaborate individual programs of follow-up and treatment. A geriatrician could request additional consultations and analyses. Two years later the medical staff in the kibbutzim completed anonymous questionnaires referring to the remote results of LGA. RESULTS: A total of 121 elderly persons were included in the LGA, performed during the period 2001-2003. The main causes of patients' referral to LGA were the known geriatric syndromes, namely functional and cognitive decline, anxiety and depression, falls etc. We assessed: (1) epidemiological data: age, gender, familial state, education, number of children, place of residence, (2) clinical data: number of both geriatric syndromes and drugs, recent changes in weight, hearing, and vision, (3) functional status: activities of daily living (ADL) and instrumental activities of daily living (IADL), falls, risk of self-inflicted injury, work, hobbies, social activity, the need for familial and social support, (4) cognitive and psychosocial status including depressive symptoms and anxiety, sleep disturbances, casualties in families or among friends, changes in mood and fears. After conducting the LGA, we recommended changes in the drug treatment, nursing, rehabilitation, institutionalization, and social help, if needed. Analysis of anonymous questionnaires showed that both medical staff and elderly in kibbutzim were satisfied with LGA implemented at the old persons' residence, they noted availability and high professional levels of LGA, additionally, they reported on the successful implementation of recommendations. The method became routine in the Upper Galilee. CONCLUSION: Since CGA is a very time- and money-consuming procedure, LGA has been tested and appears to be effective in the identification of geriatric syndromes. Within one hour of assessment, a geriatrician could retrieve a lot of the relevant information that allowed him to build individual programs for follow-up, prophylactic measures, drug and rehabilitation treatment and institutionalization. PMID- 17078428 TI - [The religious orthodox family in the pediatric hemato-oncology department]. AB - BACKGROUND: Extensive research has been conducted on the subject of cultural influences on behaviors, attitudes, beliefs and interpersonal relations of patients in the medical world. Cancer is a life-threatening illness and the intense emotional reactions to it enhance the influence of subcultural behaviors and attitudes towards the illness, the treatments and towards the medical staff. AIMS: This paper aims to describe what happens at the encounter between the medical subculture, which is mainly secular, and the orthodox subculture, of the child treated for cancer and his family. METHOD: A support group of orthodox mothers was organized. The discussions were recorded and analyzed with "content analysis" method. This article presents the themes that evolved from the analysis, which relate to the effects of the cultural encounter. RESULTS: Themes of the discussions describe the way the child and his family cope with the illness during hospitalization and at home between hospitalizations. Specific difficulties of fathers, mothers, children and adolescents become apparent. Reference was made to the dilemma of doctor vs. Rabbi, two authorities managing the treatment process. CONCLUSIONS: When the medical staff is aware and sensitive to the unique difficulties of the orthodox child and his family, this will assist in coping with the pain of the illness. Similarly, awareness of the orthodox community and leadership to the hospital culture will also help the family in its ordeal. PMID- 17078429 TI - [Treatment of pediatric pain and anxiety--the future is already here]. AB - Children may experience pain when treated in almost every field of medicine; in the Emergency department, the admission ward, or in the ambulatory clinic. Pain assessment and treatment should be an integral part of the practice of pediatrics; all of us should adopt the advocacy role to ensure humane and competent treatment of pain and suffering in children. Failure to provide adequate treatment for pain can be equated to substandard and unethical medical practice. This editorial emphasizes the progress that has been made in the research of pediatric pain and its projection on the clinical field. PMID- 17078430 TI - [The risks of combining medicine and herbal remedies]. AB - The risks of using herbal remedies, considered 'natural', should not be disregarded, as some have serious side effects and some interact with and influence conventional medical therapeutics. The effect may be pharmacokinetic by altering absorption or metabolism, and may be pharmacodynamic, by changing the final effect of the drug. St. John's wort, for example, an antidepressant herbal remedy, may pharmacodynamically interact with specific serotonin reuptake inhibitors (SSRI's), causing a serotonin syndrome. St. Johns wort also causes serious pharmacokinetic interactions by activating the cytochrome CYP3A4, dangerously decreasing blood levels of cyclosporin, warfarin, and theophylline, and reducing the efficacy of contraceptive pills and AIDS therapy. The article presents a review of a number of herbal remedies, commonly used in Israel, that have documented drug interactions, providing details of common indications, adverse reactions and drug interactions of each herbal remedy. Physicians should recognize the fact that patients use herbal remedies, purchased directly at pharmacies or health stores, and be aware of the potential interactions of these remedies with conventional drugs. PMID- 17078431 TI - [Exercise and the male reproductive system]. AB - In contrast to the effect of exercise training on the menstrual cycle in female athletes, the effect of physical activity on the male reproductive system is described far less extensively in the scientific literature. The male reproductive system consists of the hypothalamic-pituitary unit and the testes. The testes are responsible for the production of sperm and androgens, mainly testosterone. Androgens are responsible for the development of secondary male characteristics, muscle and bone growth, production of red blood cells, sex drive and other behavioral aspects. The effect of physical activity on the male reproductive axis depends on the intensity and duration of the activity, the fitness level of the individual, and his nutritional status. A single bout of short and intensive aerobic and anaerobic exercise usually increases serum testosterone level. Prolonged moderate to intense exercise (> 2 hours) leads to an initial increase followed by a decline to or below the baseline levels. Exercise training frequently results in a decrease of serum testosterone, and may rarely be associated with reduced libido, sperm production and fertility. In addition, the reduced testosterone levels may attenuate the exercise-associated muscle hypertrophy, reduce the repair of muscle damage, reduce post exercise muscle rehabilitation and may play an important role in the development of over training syndrome. Other consequences include decreased bone density and a possible effect on mood and behavior. Surprisingly, even in the medically well supervised elite athletes, changes in testosterone levels and their effects on performance and health are rarely evaluated. PMID- 17078432 TI - [Human immunodeficiency virus in newborn of infected mothers: pregnancy, breast feeding and prevention]. AB - The worldwide pandemic of human immunodeficiency virus (HIV) casts its shadow on childbirth, especially in developing countries, where the estimated rate of vertical transmission is 15%-40%, compared with 15%-25% in developed countries. To date, a total number of 90 children younger than 12 years of age are infected with the virus in Israel, and during the years 2001-2003, an average of 29 mothers living with AIDS gave birth annually. Mother-to-child transmission (MTCT) of HIV can occur during pregnancy, in the intrapartum period or postnatally, through breastfeeding. One third of all the breastfed newborn acquire the infection postnatally. HIV can penetrate through the damaged gastrointestinal mucosa into the newborn blood system. Risk increases during gastrointestinal infections and oral skin lesions. MTCT rate is 3.2 to 100 child years, especially if the mother is infected just before or after labor. Antiretroviral therapy during pregnancy and labor, and throughout the first 6 weeks of life in newborn may reduce MTCT by 3-folds. Further avoidance of breastfeeding lowers MTCT to 1% 2%. Most HIV infected women who gave birth in Israel originated from endemic countries. As breastfeeding is the preferred option in traditional cultures, early waning and alternative strategy of supplying substitutes is unique. This recommendation should be combined with medical follow-up for both the mother and her offspring, as well as psychosocial intervention, if needed. PMID- 17078433 TI - [The minimental state examination--an up-to-date review]. AB - The Mini-Mental State Examination (MMSE) was developed in 1975 as a brief test for the quantitative assessment of cognitive impairment in adults. It has since been widely used by physicians throughout the world. Its main use is for screening for cognitive impairment, which is a significant cause of morbidity and mortality in the elderly. The MMSE is also used for follow-up of cognitive changes in patients suffering from dementia and as a research instrument for assessing the efficacy of drug treatment in Alzheimer's disease. The MMSE examines orientation, immediate and short-term memory, attention and calculation, language and praxis. Construct validity of the test is considered good. An MMSE score of < or =23 or less is generally accepted as indicating cognitive impairment and was associated with the diagnosis of dementia in at least 79% of cases. The major variable that affects the MMSE's sensitivity is the level of cognitive impairment. The attainment of high levels of sensitivity increases with increased impairment. Specificity was found to be between 80-100%. Disadvantages of the MMSE include difficulty to identify mild cognitive impairment and difficulty in recording changes in cases of severe dementia. Furthermore, age, education, cultural and socioeconomic background can cause a considerable bias in the MMSE's scores. This paper aims to review the MMSE, highlight advantages and disadvantages, and describe possible clinical applications of the test. PMID- 17078434 TI - [Generic drug substitution]. AB - The rapidly rising health care expenditures, attributed mainly to the high cost of prescription drugs, have led governments around the world to look to generics as a means of containing costs in the pharmaceutical market. Generic drugs provide a less expensive alternative to brand name drugs due to the elimination of the need to perform lengthy and costly clinical trials, as required for innovative drugs. Essentially, generic substitution of drugs may be performed only after showing unequivocally that the generic formulation is identical in its active ingredients, strength, and route of administration as its innovative counterpart, and that they are bioequivalent to each other. Although the two are in essence the same, generic substitution is occasionally a controversial matter. PMID- 17078435 TI - The therapeutic effects of electrical acupuncture and auricular-plaster in 32 cases of chronic fatigue syndrome. AB - OBJECTIVE: To observe the therapeutic effects of electrical acupuncture and auricular-plaster therapy for chronic fatigue syndrome (CFS). METHOD: 64 CFS patients were randomly divided into two groups. 32 cases in the treatment group were treated by the electrical acupuncture and auricular-plaster therapy, and 32 cases in the control group with oral hydrocortisone. RESULTS: The total effective rates were respectively 93.75% in the treatment group and 75.00% in the control group, with a statistically significant difference between the two groups (P < 0.05). CONCLUSION: Electrical acupuncture and auricular-plaster therapy may show a better anti-fatigue effect than that of routine Western drugs. PMID- 17078436 TI - Point application of medicated cakes for treatment of asthma in children--a report of 60 cases. AB - OBJECTIVE: To evaluate the therapeutic effects of point application of medicated cakes for prevention and treatment of recurrent asthma in children. METHODS: Point application of medicated cakes was performed in 60 asthmatic children at the remission stage, with its effect compared with that of ketotifen used in the control group of 30 cases. RESULTS: With a total effective rate of 85%, the point application of medicated cakes was found to be superior to that of the control group (P < 0.05). CONCLUSION: Point application of medicated cakes is a simple, effective and safe therapy for asthmatic children. PMID- 17078437 TI - Treatment of cholelithiasis by acupuncture and oral decoction. AB - Thirty-six cases of cholelithiasis were treated by acupuncture at the Back-Shu and Front-Mu points plus oral decoction. The total effective rate was 97.2%, and the cured plus markedly effective rate was 83.3%, which were much better than those of 83.3% and 52.8% in the control group of 36 cases treated with oral decoction alone. The statistical differences between the two groups were respectively P < 0.05 and P < 0.01. PMID- 17078438 TI - Acupuncture treatment of generalized anxiety disorder. PMID- 17078439 TI - Acupuncture treatment for 68 cases of functional impairment induced by cerebral hemorrhage at the convalescence stage. PMID- 17078440 TI - Acupuncture and cupping for treatment of hiccup in cases of cerebrovascular accident. AB - OBJECTIVE: To observe the therapeutic effects of acupuncture and cupping for hiccup in cases of cerebrovascular accident. METHOD: 80 cases of hiccup due to cerebrovascular accident were randomly divided into the treatment group of 40 cases treated by acupuncture and cupping and the control group of 40 cases treated with Ritaline. RESULT: In the treatment group, 24 cases were cured, 8 cases markedly effective, 5 cases improved, and 3 cases failed, with a total effective rate of 92.5%. In the control group, 9 cases were cured, 12 cases markedly effective, 8 cases improved, and 11 cases failed, with a total effective rate of 72.5%. There was a statistically significant difference in the therapeutic effects between the two groups (mu = 3.3259, P = 0.0009). CONCLUSION: The effect of acupuncture and cupping for hiccup due to cerebrovascular accident was noticeably superior to Ritaline. PMID- 17078441 TI - Acupuncture treatment of Jacksonian epilepsy--a report of 98 cases. AB - OBJECTIVE: To evaluate the effect of acupuncture treatment for controlling Jacksonian epilepsy. METHOD: Penetrating needling is adopted together with scalp acupuncture and strong/electric needling on body points. RESULTS: Of the 98 cases treated for 5 courses, 52 cases were markedly effective, 22 effective, 10 poorly effective and 14 ineffective. The total effective rate was 85.7% (95% CI = 78.8% 92.6%). CONCLUSION: Acupuncture is quite effective for treating Jacksonian epilepsy. PMID- 17078442 TI - Sixty-four cases of scapulohumeral periarthritis treated by auricular plaster therapy. PMID- 17078443 TI - Treatment of the localized neurodermatitis by plum-blossom needle tapping and with the modified yangxue dingfeng tang--a clinical observation of 47 cases. AB - OBJECTIVE: To observe the therapeutic effects of plum-blossom needle tapping combined with the modified yangxue dingfeng tang (a decoction for nourishing the blood and expelling wind) for the localized neurodermatitis. METHODS: 141 cases of the localized neurodermatitis were randomly divided into 3 groups. Forty-seven patients in the treatment group received local tapping and oral medication of the modified yangxue dingfeng tang which consisted of sheng di huang ([Chinese characters: see text] Radix Rehmanniae), dang gui ([Chinese characters: see text] Radix Angelicae Sinensis), chi shao ([Chinese characters: see text] Radix Paeoniae Rubra), tian dong ([Chinese characters: see text] Radix Asparagi), mai dong ([Chinese characters: see text] Radix Ophiopogonis), jiang can ([Chinese characters: see text] Bombyx Batryticatus), mu dan pi ([Chinese characters: see text] Cortex Moutan), chuan xiong ([Chinese characters: see text] Rhizoma Chuanxiong), he shou wu ([Chinese characters: see text] Radix Polygoni Multiflori), and quan xie ([Chinese characters: see text] Scorpio). Forty-seven patients in the control group I were simply given the modified yangxue dingfeng tang; and 47 patients in the control group II were treated with oral benadryl and Vitamin C plus local external application of 10% urea ointment. The treatment course for all the 3 groups lasted 30 days. RESULTS: The short-term and long-term effects for the treatment group were much better than those of the 2 control groups (P < 0.05). CONCLUSION: The plum-blossom needle tapping plus the modified yangxue dingfeng tang exhibits a better and stable effect for the localized neurodermatitis. PMID- 17078444 TI - Thirty-six cases of pseudobulbar palsy treated by needling with prompt and deep insertion. PMID- 17078445 TI - TCM treatment for protrusion of lumbar intervertebral disc--a report of 100 cases. PMID- 17078446 TI - The effect of acupuncture on serum IgE level in patients with chronic urticaria. AB - In order to explore the effect of acupuncture on serum IgE level in patients with chronic urticaria, the changes of the immune indices were observed at various stages in the development of the disease. It was found that the serum IgE level in patients with chronic urticaria was higher than that of the normal healthy subjects; and that acupuncture had a benign regulatory effect on IgE, showing a favorable regulation on the immune functions in patients with chronic urticaria. PMID- 17078447 TI - Clinical observation on acupuncture treatment of depressive neurosis in 30 cases. PMID- 17078448 TI - He-Ne laser auricular irradiation plus body acupuncture for treatment of acne vulgaris in 36 cases. AB - In order to observe the therapeutic effects of He-Ne laser auricular irradiation plus body acupuncture for acne vulgaris, 68 cases of acne vulgaris were randomly divided into a treatment group of 36 cases treated with He-Ne laser auricular irradiation plus body acupuncture, and a control group of 32 cases treated with body acupuncture only. The results showed that the cure rate was 77.8% in the treatment group and 46.9% in the control group (P < 0.05), indicating that He-Ne laser auricular irradiation plus body acupuncture may exhibit better effects for acne vulgaris. PMID- 17078449 TI - Forty-six cases of hysteria treated mainly by acupuncture at Siguan points. PMID- 17078450 TI - The therapeutic effects of point massage for angina pectoris in 30 cases of coronary heart disease. PMID- 17078451 TI - Treatment of psoriasis vulgaris by oral administration of yin xie ping granules- a clinical report of 60 cases. PMID- 17078452 TI - Forty cases of tumorous fever treated with TCM method of removing blood heat and dissipating blood stasis. PMID- 17078453 TI - Dr. Sun Shidao's experience in TCM treatment of acne. PMID- 17078454 TI - Dr. Feng Xinghua's experience in treating ankylosing spondylitis. PMID- 17078455 TI - Dr. Yi Juqing's experience in treating female infertility. PMID- 17078456 TI - Effects of long-term use of the heat-clearing, diuresis-promoting and collateral mediating chinese drugs on changes of proteinuria in patients with chronic nephritis. AB - PURPOSE: To observe and evaluate the effects of long-term use of the heat clearing, diuresis-promoting and collateral-mediating Chinese drugs on changes of proteinuria in patients with chronic nephritis, to analyze the influence of renal function, blood pressure, proteinuria and TCM syndrome types on therapeutic effects of the therapy, and to analyze the indications and effects of the heat clearing, diuresis-promoting and collateral-mediating Chinese drugs when added with small dosage of Lei Gong Teng ([Chinese characters: see text] Radix et Rhizoma Tripterygii). METHOD: 43 cases of chronic nephritis were treated for one year with the heat-clearing, diuresis-promoting and collateral-mediating Chinese drugs. RESULT: A one-year treatment showed obvious reduction of proteinuria in patients with chronic nephritis. CONCLUSIONS: (1) The heat-clearing, diuresis promoting and collateral-mediating Chinese drugs are effective for a long-term treatment of proteinuria in patients with chronic nephritis. (2) Additional use of Lei Gong Teng can help control proteinuria. (3) Damp-heat deeply accumulated in the kidney is one of the pathologic characteristics for chronic nephritis with the damp-heat syndrome. PMID- 17078457 TI - Influence of organ pathology on the electrical parameters in organ projection areas of the skin. AB - Pathology of an internal organ causes significant rectification of electrical currents (diode phenomenon) in related skin areas--organ projection areas (OPAs), once the resistance 'breakthrough effect' has been induced in the skin. Also, the impedance of the diseased organ's projection areas is increased. The aim of this double-blind study was to confirm these phenomena statistically using a broad spectrum of measuring parameters. Skin impedance vs. measurement frequency, skin impedance vs. applied voltage, and skin resistance vs. voltage were evaluated at 335 auricular OPAs related to the healthy internal organs and 203 auricular OPAs related to the diseased organs. These measurements confirmed that the skin electrical characteristics of specific locations are dependent on the health state of the corresponding internal organs. It was found that the impedance of OPAs corresponding to the healthy internal organs equals 185 +/- 291 komega at 10 Hz and decreases to 64 +/- 49 komega at 10 kHz; and that the impedance of OPAs related to the diseased organs equals 7.5 +/- 2.2 Momega at 10 Hz and 99 +/- 50 komega at 10 kHz. The disparity of the impedances between the healthy and diseased organs related OPAs is greater at higher potentials. The skin resistance, when measured with a negatively polarized point electrode, undergoes a rapid resistance decrease of approximately two orders of magnitude, if the applied current is sufficient. After this reversible 'breakthrough effect' is obtained the skin exhibits rectification. The degree of rectification is low for OPAs related to the healthy organs. For OPAs related to the diseased organs, the resistance measured with a positive polarisation of measuring electrode could be five times greater than the resistance measured at the same voltage with the same but negatively polarised electrode. The disparity between the resistance measured with a negatively and with a positively polarised electrode at OPAs related to the diseased organs was greater at higher measurement voltages. The influence of organ pathology on the electrical parameters of related OPAs does not depend on the kind of internal organ and is not affected by the etiology of pathology. The degree of rectification or difference in impedance is proportional to the extent of the pathological process within the related organ. PMID- 17078458 TI - How to treat nocturnal enuresis in children? PMID- 17078459 TI - Acupuncture treatment of hypochondriac pain. PMID- 17078460 TI - A brief history of human constitutionology. AB - Human constitutionology studies human itself, which was proposed by Dr. Kuang. The study on human constitution in Chinese medicine has ever increased since the late 1970s. This review on the development of human constitutionology in Chinese Traditional Medicine (TCM) and western medicine is expected to give people with a different background to understand the establishment of this new subject, and its significance in promoting modernization of TCM and multi-disciplinary collaboration in human study and as an individualized model guiding the re orientation of modern medicine. PMID- 17078461 TI - A TCM theoretical study on the etiology and pathogenesis of metabolic syndrome. PMID- 17078462 TI - Mite allergens avoidance in asthma: what we need to do? PMID- 17078463 TI - Comparison of two specific allergen screening tests in different patient groups. AB - UniCAP and multiple-antigen simultaneous test (MAST) are among the widely used specific allergen tests. The Phadiatop and Fx5 are the multi-allergen UniCAP screening tests for inhalant allergens and common food allergens, respectively. We compared the differences between Phadiatop-Fx5 and MAST as screening allergen tests to clarify the optimal tests for different groups of allergic patients. Serum samples of 224 consecutive allergic patients were tested with Phadiatop, Fx5 and MAST. Results of these allergen tests were compared and analyzed in subgroups categorized by age, serum IgE levels and the clinical departments where the patients were treated. We found that among the 224 patients, 155 patients (69.2%) tested positive with Phadiatop while 137 (61.1%) tested positive with MAST for inhalant allergens. Twenty patients were Phadiatop(+)/MAST(-), while only 2 were Phadiatop(-)/MAST(+). There were 57 patients (25.4%) who tested positive with Fx5, while 32 (14.3%) tested positive with MAST for food allergens. Thirty-eight patients were Fx5(+)/MAST(-), while 13 were Fx5(-)/MAST(+). The disagreement between these two tests was more apparent in food allergen tests than in inhalant allergen tests. Most cases of disagreement occurred in younger age groups and in the patient group with IgE > 500 IU/ml. These results suggested that UniCAP allergen screening tests might be more effective in certain patient groups as screening tests. PMID- 17078464 TI - Lamotrigine adjunctive therapy in children with refractory epilepsy: a medical center study. AB - The aim of this study was to investigate the effects of adjunctive lamotrigine (LTG) in refractory epilepsy in Taiwan, especially including patients who had undergone ineffective epileptic surgeries. Totally, 27 recruited children and adolescents with refractory epilepsy were treated with LTG add-on therapy, but 23 were enrolled statistically during 2 years period trial. Ten of the 23 patients (43.5%), including 5 undergoing epileptic surgeries, before had a > 50% reduction in seizure frequency. Surprisingly, LTG was found to have excellent response in six patients with Lennox-Gastaut Syndrome (LGS), 5 of whom had received anterior 2/3 callosotomy prior to the trial. Non-seizure related positive effects were presented in 3 patients, including greater alertness and attentiveness in 2 patients and improvement in mood in one patient. This open trial found that LTG was an effective add-on anti-epileptic drug (AED) in Taiwanese children and adolescents with refractory epilepsy and in cases with LGS even after ineffective callosotomy. PMID- 17078465 TI - Distribution of allergens in children with different atopic disorders in central Taiwan. AB - Allergic disorders, including asthma, allergic rhinitis, and eczema, are the most common chronic childhood diseases. Exposure to house dust mites (HDMs) can exacerbate allergic disorders in sensitized individuals. The data for sensitization to HDMs and other frequent allergens amongst atopic children in Taiwan is limited. We studied 498 children (aged 2-16 years) with atopy in central Taiwan with CAP testing (Cationic Antimicrobial Protein system). Our results revealed a high prevalence of sensitization to Dermatophagoides pteronyssinus (Der p) (90.2%), Dermatophagoides farinae (Der f) (88.2%), Dermatophagoides microceras (Der m) (79.5%), and Blomia tropicalis (Blo t) (76.7%) amongst the children. In contrast to HDM, the sensitization rates for other aeroallergens including cockroaches, dog dander, cat dander, Aspergillus fumigatus, Candida albicans, Cladosporium herbarum, Penicillium notatum were not common among the study children. With respect to age, inhaled allergen sensitization predominated in older children, whereas the inverse occurred with food allergens. In addition, a relatively higher proportion of co-sensitization between Der m and the other three antigens, including Der p, Der f, Blo t was found. Our results suggest that HDMs, including Der p, Der f, Der m, and Blo t allergens, act as important inducers of symptoms in Taiwanese allergic children. PMID- 17078466 TI - Congenital mesoblastic nephroma presenting with massive hematuria and hemorrhagic shock: report of one case. AB - Congenital mesoblastic nephroma (CMN) is a rare benign tumor that occurs during the neonatal period and early infancy. The vast majority of these tumors present as asymptomatic palpable abdominal masses. We describe an unusual presentation of a CMN in a 10-month-old male infant who presented with massive hematuria and the development of hemorrhagic shock. Abdominal ultrasound showed a heterogeneous solid complex mass measuring 4.8 x 3.5 cm arising from the upper pole of the left kidney. The patient was resuscitated using intravenous fluids and blood transfusions because persistent massive bloody urine leading to progressive shock occurred the night of the admission day. Preoperative diagnosis was possible Wilms tumor of the left kidney. The histopathological findings were consistent with the character of a cellular variant of CMN. The patient was free of recurrence and metastasis at the 2-year follow-up examination. Our case report suggests that CMN is a rare benign renal tumor during infancy and may present with unusual massive hematuria and shock. PMID- 17078467 TI - Pallister-Killian syndrome: report of one case. AB - Pallister-Killian Syndrome (PKS) is a rare sporadic congenital anomaly disorder, characterized by multiple congenital anomalies, especially craniofacial dysmorphism. It is also associated with mental retardation, seizure, skin pigmentation, and visceral malformations such as congenital diaphragmatic hernia, congenital heart defect, anorectal anomalies, and genital malformation. This syndrome usually presents with tissue-limited mosaicism of supernumerary 12p isochromosome i (12p). Moreover, diagnosis of Pallister-Killian Syndrome (PKS) is difficult because the ratio of abnormal to normal karyotyping is much lower in peripheral lymphocytes than in skin fibroblasts. We report the first case in Taiwan, who has tetrasomy 12p mosaic in peripheral lymphocytes. PMID- 17078468 TI - Goldenhar syndrome (oculoauriculovertebral dysplasia): report of one case. AB - Goldenhar syndrome, also known as oculoauriculovertebral dysplasia, is an uncommon condition, characterized by a combination of anomalies: epibulbar dermoids or lipodermoids, preauricular appendices, malformation of the ears, hemifacial microsomia, vertebral anomalies, and others. The etiology of this disease has remained unclear; factors including chromosomal abnormalities, maternal diabetes mellitus or drug use, and influence of environment during pregnancy have been proposed. Here, we describe a case of Goldenhar syndrome in a 1-day-old female newborn, who presented with right external ear atresia, left preauricular appendices, cleft-like extension of the right oral angle, mandibular hypoplasia and relatively small hands. The literature on Goldenhar syndrome is briefly reviewed. PMID- 17078469 TI - Synchronous multifocal osteosarcoma: report of one case. AB - Synchronous multifocal osteosarcoma (SMOS), defined as more than one bone lesion at presentation, is a rare variant form of osteosarcoma. The onset is usually in childhood or early adolescence without pulmonary metastasis. The prognosis has been dismal. Whether SMOS represents a true multicentic origin or merely bone-to bone metastases remains controversial. Here, we report a case of SMOS in a 10 year-old girl, with the dominant primary sclerotic tumor arising from the right distal femur. Despite aggressive chemotherapy and limb salvage surgery, she died of progressive multiple axial skeletal and symmetrical metaphyseal long bone diseases within one year after diagnosis. No pulmonary metastasis was found before she died. PMID- 17078470 TI - Pathological fracture as a manifestation of extramedullary blastic crisis in chronic myelogenous leukemia: report of one case. AB - A three-year-old girl with chronic myelogenous leukemia (CML) experienced a pathological fracture of her femur after a demonstrated osteolytic bone lesion. Extramedullary disease (EMD) was diagnosed following the histologic findings of a biopsy of the osteolytic lesion. Frank blast crisis in the bone marrow followed one month later. This was the youngest patient to have been reported in English literature of Philadelphia chromosome positive (Ph+) CML with isolated bony EMD and pathological fracture. Treatment with a tyrosine kinase inhibitor, imatinib mesylate (Gleevec), in bone marrow accelerated phase of CML was failed to reverse the progression of blastic transformation, neither in the extramedullary bone lesion nor in the bone marrow. PMID- 17078471 TI - Detecting bacterial contamination in platelet products. AB - Bacterial contamination of platelets is an important cause of transfusion associated morbidity and mortality. It is currently the most frequent infectious complication of transfusion therapy, with between 1 in 1,000 and 1 in 3,000 platelet units being bacterially contaminated at time of transfusion. Several factors have contributed to the persistence of this problem including lack of sensitive detection methods, lack of recognition of the frequency of the problem, inadequate recognition of septic reactions by clinicians treating patients receiving platelet transfusions, differences in transfusion reactions between bacterial species and bacterial inocula transfused, and differing methodologies and time of testing for detection of bacteria in platelet units. There are also important correlations between the receipt of bacterially contaminated platelet units and the development of transfusion reactions and bacteremia. In the last few years the recognition of the importance of platelet bacterial contamination prompted the College of American Pathologists (CAP) and the American Association of Blood Banks (AABB) to set new standards requiring the screening of platelets for bacterial contamination. In the wake of these standards, an increasing number of approaches have been and are being developed to deal with this problem. The clinical sensitivity, specificity and predictive value of these detection methods vary considerably and need to be defined for routine laboratory practice. In this review, we focus on the practical aspects and feasibility of implementing FDA cleared detection methods for identifying bacterially contaminated platelet units. We also present details of a number of methods under development for at issue use. PMID- 17078472 TI - Preanalytic indicators of laboratory performances and quality improvement of laboratory testing. AB - Laboratory diagnosis is traditionally a three-part process that develops within the preanalytic, analytic and post-analytic phases. There is consolidated evidence that lack of standardization and monitoring of preanalytic variables, including procedures for patient identification, sample collection, handling and processing has an adverse influence on the reliability of test results, consuming valuable healthcare resources and compromising the patient's outcome. The preanalytic phase enfolds the greatest potential for quality improvement, once reliable strategies are identified and applied. A comprehensive quality program should outstrip the traditional confines of clinical laboratories, encompassing reliable monitoring policies of the state of quality across the entire process. Such an approach requires the adoption of a reliable global quality monitoring system based on a core set of broad, evidence-based preanalytic performance measures. The present article synthesizes current evidence on this topic, defining a tentative approach for implementation of a preanalytic quality monitoring system in clinical laboratories. PMID- 17078473 TI - Multicenter evaluation of a new 4th generation HIV screening assay Elecsys HIV combi. AB - Fourth-generation screening assays which permit a simultaneous detection of human immunodeficiency virus (HIV) antigen and antibody reduce the diagnostic window on average by four days in comparison to third-generation antibody assays. Recently, the new automated Elecsys HIV combi was compared in a multicenter study to alternative fourth- and third-generation assays, p24 antigen test and HIV-1 RNA RT-PCR. A total of 104 serocon-version panels, samples of the acute phase of infection after seroconversion (n = 33), anti-HIV-1 positive specimens (n = 572) from patients in different stages of the disease, 535 subtyped samples from different geographical locations, including group M (subtypes A-J) and group O, anti-HIV-2 positive sera (n = 364), dilutions of cell culture supernatants (n = 60) infected with different HIV-1 subtypes, selected performance panels, 8406 unselected samples from blood donors originating from different blood transfusion centers, 3810 unselected sera from daily routine and from hospitalized patients, 9927 unselected samples from South Africa and 1943 potentially interfering samples were tested with the Elecsys HIV combi. Elecsys HIV combi showed a comparable sensitivity to HIV-1 Ag stand-alone assays for early detection of HIV infection in seroconversion panels. The mean time delay of Elecsys HIV combi (last negative sample + 1 day) in comparison to HIV-1 RT-PCR for 92 panels tested with both methods was 3.23 days. The diagnostic window was reduced with Elecsys HIV combi between 1.56 and 5.32 days in comparison to third-generation assays. The specificity of Elecsys HIV combi in blood donors was 99.80% after repeated testing. Our results show that a fourth-generation assay with improved specificity and sensitivity like the Elecsys HIV combi is suitable for blood donor screening due to its low number of false positives and since it detects HIV p24 antigen with a comparable sensitivity to single antigen assays. PMID- 17078474 TI - Hereditary spherocytosis and other hemolytic anomalies distort diabetic control by glycated hemoglobin. AB - Glycated hemoglobin (%HbA1c) has become a well established and reliable long term parameter indicative of the mean glucose load of the preceding 8 -10 weeks. A normal life span of approximately120 days of the red blood cells (RBC) is an essential condition. Hemolytic affections are characterized by a shorter life span, reducing the overall glucose uptake and %HbA1c. Measured %HbA1c is no longer correlated with mean blood glucose of the preceding period, simulating false low values. Underestimation of this kind is demonstrated for several hemolytic conditions, among them hereditary spherocytosis (HS). This latter, often harmless anomaly may lead to serious underestimation of glucose load. Recent investigations show not only a much higher incidence of HS than hitherto admitted (approximately 1 in 250 persons) but also an abnormally high incidence of diabetes in this hemolytic affection. In the presence of hemolysis %HbA1c is to be interpreted with caution. This justifies systematic detection of HS in routine hematology using--if available--the increase of the percentage of hyperchromic RBC. PMID- 17078475 TI - Comparison of estimated glomerular filtration rates from serum creatinine and cystatin C in patients with impaired creatinine production. AB - Determination of the glomerular filtration rate (GFR) is important for the drug dosage adjustment and clinical management of patients. The aim of our study was the comparison of estimated GFR values from serum creatinine (eGFRcreatinine) and cystatin C (eGFRcystatin C) in patients with impaired creatinine production. A total of 564 serum samples from patients with kidney disease (n=179), liver (n=71) and kidney (n=182) transplants, critically ill patients (n=82) and healthy subjects (n=50) were analyzed for serum creatinine and cystatin C. The creatinine production rate (CPR) was significantly lower in the different groups of patients than in the control group (p<0.001). A negative correlation was found between the eGFRcreatinine/eGFRcystatin C ratio and CPR (r= -0.964, p<0.001). For CPR higher than 800 mg/24h both procedures for estimating the GFR classified values higher and lower than 60 mL/min with an acceptable agreement; however, for CPR less than 800 mg/24h the eGFRcreatinine led to false negatives in a high number of cases with eGFRcystatin C <60 mL/min. PMID- 17078476 TI - Evaluation of direct and indirect markers to assess the androgen status in healthy males during aging. AB - Evaluation of the male androgen status requires a marker that reflects the biologically active fraction of plasma testosterone. The serum sex hormone binding globulin (SHBG) concentration is not suitable here because of its wide inter-individual scatter. As potential biological markers of the active testosterone fraction we compared indirect methods calculated on the basis of SHBG and total testosterone measured by fully automated IMMULITE 2000 assays (DPC, Los Angeles, CA, USA), and total testosterone alone with direct free testosterone measured by RIA (DPC). Indirect methods were the free androgen index FAI, calculated free testosterone cFT, and calculated bio-available testosterone cBT. Further androgens measured were DHEAS and androstenedione. Blood samples were collected from a cohort of 446 healthy men aged between 20-99 years. All parameters except SHBG decreased significantly during aging. The direct free testosterone assay was significantly correlated with the indirect androgen parameters. This is in accordance with earlier results using LC-MS as the gold standard method. The strongest correlation was seen with cBT/measured albumin (r=0.750), though the direct testosterone RIA does not measure the entire unbound fraction of testosterone, and total testosterone can rapidly be measured with an automated assay system. It was found that a fixed albumin concentration of 43 g/L is a reasonable calculation basis for cBT in subjects of <70 years. In the elderly >70 years or persons with known pathologies of the androgen axis, it is commendable to measure the albumin concentration individually. In conclusion, calculated bio-available testosterone (cBT) is the best marker to reflect the bioactive testosterone fraction, i.e. the androgen status in males. PMID- 17078477 TI - Tartrate-resistant acid phosphatase 5b (TRACP 5b) as a marker of bone resorption. AB - Tartrate-resistant acid phosphatase (TRACP) is an enzyme that is expressed in high amounts by bone resorbing osteoclasts, inflammatory macrophages and dendritic cells. Two forms of TRACP circulate in human blood, TRACP 5a derived from macrophages and dendritic cells, and TRACP 5b derived from osteoclasts. Recent data have demonstrated the utility of TRACP 5b as a marker of osteoclast number and bone resorption, and serum TRACP 5a as a marker of inflammatory conditions. This review summarizes the scientific knowledge on the role of TRACP in osteoclastic bone resorption, the mechanism of TRACP 5b generation in osteoclasts and its secretion into the blood circulation, the methodology of measuring TRACP 5b, diagnostic evidence for the use of TRACP 5b as a resorption marker, and characteristics of TRACP 5b compared to other commonly used bone turnover markers. PMID- 17078478 TI - Observation of serum erythropoietin concentrations in female athletes for up to eight days after a marathon run. AB - Erythropoietin (EPO) was studied in 13 female marathon runners before and up to 8 days after a competition marathon run. The median baseline control value was 13.7 U/l. No change in EPO concentration was found immediately (15 min.) and one day after the run. However, a median increase in EPO concentration (18.1 U/l) was found on day three post-exercise (p< 0.05). On day 8 no change was found compared to pre-exercise values. This late increase in EPO concentration would seem to be responsible for the well known increase of red blood cell mass in long distance runners. PMID- 17078479 TI - Human adenovirus type 1 related to feline adenovirus: evidence of interspecies transmission. AB - Adenovirus is recognized to be a significant global enteropathogen in association with sporadic cases as well as outbreaks of acute gastroenteritis in humans. Based on the genetic analysis, one adenovirus strain bearing feline adenovirus gene was detected in a fecal specimen collected from a 1-year old female child with acute gastroenteritis in Japan. The human adenovirus detected and feline adenovirus shared high identities (100% and 97%) at the amino acid levels of hexon and fiber genes, respectively, and they belonged to the same human Ad1 cluster (known as the prototype Adenoid 71). These findings suggest that the interspecies transmission of adenovirus between humans and felines might occur in nature. This report is noteworthy because it is the first, to the best of our knowledge, providing evidence of adenovirus type 1 transmission between humans and animals, and highlights possible zoonoses in humans. Further epidemiological studies should be conducted to determine whether this adenovirus strain will be emergent in future. PMID- 17078480 TI - Detection and genetic characterization of norovirus strains circulating among infants and children with acute gastroenteritis in Japan during 2004-2005. AB - A total of 752 fecal specimens collected during the period of July 2004 to June 2005 from infants and children with acute gastroenteritis from four different regions (Maizuru, Tokyo, Sapporo, and Osaka) of Japan were tested for the presence of norovirus by RT-PCR. It was found that 139 (18.5%) fecal specimens were positive for norovirus. Norovirus infection was detected almost all year round with the highest prevalence in January. Norovirus GII was the most predominant genogroup (98.6%; 137 of 139). The genotypes detected in this study were GI/1, GII/1, GII/3, GII/4, and GII/6. Of these, NoV GII/4 (known as the Lordsdale virus cluster) was re-emerging and became the leading genotype (77.7%). Meanwhile, the incidence of NoV GII/3 (known as the Arg320 virus cluster) has dropped rapidly, accounting for only 15.8%. Another interesting feature of the study was the identification of Picton03/AU-like recombinant NoV for the first time in Japan. Based on the genetic analysis, it was interesting to note that NoV GII/4 in 2004-2005 made a distinct cluster in comparison to other NoV GII/4 circulating in 2002-2003 and 2003-2004. Of note, "new recombinant variant designated GIIb" within NoV GII/3, which was first detected in Saga City, Japan in 2003-2004 in only one case, had increased, spreading widely in Japan and representing 45.5% (10 of 22). Further epidemiological studies should be conducted to determine whether this new recombinant variant strain will be dominant in Japan in the coming year. PMID- 17078481 TI - Total plasma homocysteine in Bulgarian population measured by high-performance liquid chromatography with fluorescence detection--comparison with fluorescence polarization immunoassay. AB - Over the last decade, evidence has accumulated that elevated total homocysteine (tHcy) is an independent risk factor for vascular disease. Due to the variety of Hcy determinants (age, gender, ethnicity and lifestyle), it is now recommended that the distribution of plasma Hcy concentrations should be established for different populations. Therefore the objective of our study was to evaluate a modified HPLC with fluorescence detection procedure for reliable quantification of tHcy and to demonstrate its successful application to determine the distribution of tHcy levels in healthy Bulgarians. The presented method showed good analytical performance (intra- and interassay CVs were <3.9% and <6.7%, respectively; inaccuracy was <6.5%, and analytical recovery 95%-98%, the detection limit was 0.3 micromol/l) and no drug interference was registered. Comparison between HPLC-FD and FPIA using Passing-Bablok regression analysis (r=0.9906) showed good agreement. We describe the distribution of plasma tHcy in a group of 162 healthy Bulgarian adults and examined its relation with age and gender. Our results indicate that higher Hcy concentrations were associated with male sex and increasing age. The higher plasma Hcy observed in our population compared to the rest of Europe corresponds to the high prevalence and mortality of cardiovascular disease in Bulgaria. PMID- 17078482 TI - Antiretroviral therapy in AIDS patients with tuberculosis. AB - Tuberculosis associated with HIV infection continues to be an important problem throughout the world. Since the advent of HAART, the medication of HIV-infected patients who have to receive concomitant treatment for tuberculosis has become a difficult task. The two main problems faced by clinicians include the significant pharmacokinetic interactions between rifamycins, a cornerstone in antituberculosis therapy, and protease inhibitors and nonnucleoside reverse transcriptase inhibitors, which are essential components of antiretroviral combination regimens, as well as the best moment to initiate antiretroviral therapy in patients with tuberculosis. The therapy of choice for patients with no previous antiretroviral experience includes an antituberculous regimen with rifampin and an efavirenz-based antiretroviral regimen. No dose adjustments of these drugs seem to be necessary. Nevirapine can be an alternative to efavirenz in this situation. For patients who cannot take efavirenz, either due to resistance or intolerance, rifabutin and a boosted protease inhibitor can be coadministered, with the necessary dose adjustments. No definite recommendations can be given regarding the optimal timing of antiretroviral therapy, but a delay of two months after initiation of antituberculosis therapy would be advisable and seems to be safe in most patients. PMID- 17078483 TI - HIV evolutionary dynamics within and among hosts. AB - The HIV evolutionary processes continuously unfold, leaving a measurable footprint in viral gene sequences. A variety of statistical models and inference techniques have been developed to reconstruct the HIV evolutionary history and to investigate the population genetic processes that shape viral diversity. Remarkably different population genetic forces are at work within and among hosts. Population-level HIV phylogenies are mainly shaped by selectively neutral epidemiologic processes, implying that genealogy-based population genetic inference can be useful to study the HIV epidemic history. Such evolutionary analyses have shed light on the origins of HIV, and on the epidemic spread of viral variants in different geographic locations and in different populations. The HIV genealogies reconstructed from within-host sequences indicate the action of selection pressure. In addition, recombination has a significant impact on HIV genetic diversity. Accurately quantifying both the adaptation rate and the population recombination rate of HIV will contribute to a better understanding of immune escape and drug resistance. Characterizing the impact of HIV transmission on viral genetic diversity will be a key factor in reconciling the different population genetic processes within and among hosts. PMID- 17078484 TI - Do regulatory T-cells play a role in AIDS pathogenesis? AB - The impairment of adaptive immune responses to HIV and abnormalities in the immune regulatory function mechanisms during HIV infection have been regarded as key issues in AIDS pathogenesis since the early years of the pandemic. However, the multiple mechanisms underlying this impairment are still not fully understood. New emerging information shows that alterations in the number and/or function of regulatory T-cells may contribute to HIV pathogenesis. Thus, pharmacologic manipulation of regulatory T-cells as well as blocking the activity of other immunomodulatory molecules, such as indoleamine 2,3-dioxygenase, glucocorticoid-induced tumor necrosis factor receptor and PD1, might provide a valuable approach to redirect the immune system towards an efficient antiviral response. PMID- 17078485 TI - Human APOBEC3 proteins, retrovirus restriction, and HIV drug resistance. AB - Over 40 million people worldwide currently have HIV/AIDS. Many antiretroviral drugs have proven effective, but drug-resistant HIV variants frequently emerge to thwart treatment efforts. Reverse transcription errors undoubtedly contribute to drug resistance, but additional significant sources of viral genetic variation are debatable. The human APOBEC3F and APOBEC3G proteins can potently inhibit retrovirus infection by a mechanism that involves retroviral cDNA cytosine deamination. Here we review the current knowledge on the mechanism of APOBEC3 dependent retrovirus restriction and discuss whether this innate host-defense system actively contributes to HIV genetic variation. PMID- 17078486 TI - Reproductive options for HIV-serodiscordant couples. AB - The foundation of a family is a personal enterprise with deep biological and emotional motivations. Family life is a no lesser priority for individuals living with HIV than for others. Moreover, the dramatic reduction in HIV-related morbidity and mortality as a consequence of the widespread use of HAART has led to a growing number of HIV-infected individuals and their partners requiring education and counseling regarding HIV disease and reproduction. After careful evaluation of health status and fertility in both partners, two main alternatives may be considered--natural conception or assisted reproduction techniques. This review discusses the pros and cons of reproductive options in HIV-infected persons and proposes a protocol for their counseling. PMID- 17078487 TI - [Renal replacement therapy in the selected connective tissue diseases]. PMID- 17078488 TI - [In patients with Graves' disease signal-averaged P wave duration positively correlates with the degree of thyrotoxicosis]. AB - Coexistence of the goitre, proptosis and palpitations was observed in XIX century for the first time. Sinus tachyarytmias and atrial fibrillation are typical cardiac symptoms of hyperthyroidism. Atrial fibrillation occurs more often in patients with toxic goiter than in young patients with Grave's disease. These findings suggest that causes of atrial fibrillation might be multifactorial in the elderly. The aims of our study were to evaluate correlations between the parameters of atrial signal averaged ECG (SAECG) and the serum concentration of thyroid free hormones. 25 patient with untreated Grave's disease (G-B) (age 29,6 +/- 9,0 y.o.) and 26 control patients (age 29,3 +/- 6,9 y.o.) were enrolled to our study. None of them had history of atrial fibrillation what was confirmed by 24-hour ECG Holter monitoring. The serum fT3, fT4, TSH were determined in the venous blood by the immunoenzymatic method. Atrial SAECG recording with filtration by zero phase Butterworth filter (45-150 Hz) was done in all subjects. The duration of atrial vector magnitude (hfP) and root meat square of terminal 20ms of atrial vector magnitude (RMS20) were analysed. There were no significant differences in values of SAECG parameters (hfP, RMS20) between investigated groups. The positive correlation between hfP and serum fT3 concentration in group G-B was observed (Spearman's correlation coefficient R = 0.462, p < 0.02). No significant correlations were found between RMS20 and serum fT3 in G-B group and between hfP or RMS 20 and serum fT3 in group K. These findings suggest that occurrence of atrial fibrillation in patients with Grave's disease depends not only on hyperthyroidism but on serum concentration of fT3 also. PMID- 17078489 TI - [Analysis of some risk factors of coronary artery calcification in peritoneal dialysis patients]. AB - Cardiovascular disease (CVD) is a major cause of death in peritoneal dialysed patients (PD-pts). Coronary artery calcification (CAC) is likely to affect the development of CVD. Purpose of our study was to evaluate coronary artery calcification and risk factors of this calcification in PD-pts. We studied 62 patients (38 F, 24 M) undergoing peritoneal dialysis (PD). Coronary calcification was examined by ECG-gated multidetector CT (Light Speed Ultra) using Agatson (AG) and volumetric (V) methods. Patients were divided into 3 groups depending on mean value of estimated CAC: group A-no calcification, group B-CAC maximal value 400 mm3, group C-CAC value more than 400 mm3. As risk factors of CAC were evaluated: patients age, sex, dialysis duration, serum concentration of Ca, P, homocysteine CRP and fibrinogen, as well as, CaxP product, intact PTH; presence of diabetes or hypertension. Coronary artery calcification was detected in 68% of patients. In the whole observed population positive correlation between CAC determined by AG and V methods and CRP (r = 0.36, p < 0.05) as well as patients age (r = 0.5, p < 0.01) was observed. There was also positive correlation between CAC and fibrinogen concentration (AG CAC r = 0.58, p < 0.05; V CAC r = 0.72, p < 0.05). When compared group C with the groups A and B cardiovascular complications were in this group more frequent than in the last two: 4 patients from group C died because of cardiovascular complications. PMID- 17078490 TI - [Effect of the optimal neurohormonal blockade on long-term survival in patients with chronic heart failure]. AB - BACKGROUND: In recent years large scale clinical trials have cleary shown that a number of pharmacological treatments can improve the outcomes of patients (pts) with chronic heart failure (CHF). AIM: The aim of this study was to assess the effect of optimal neurohormal blockade in pts with chronic heart failure on survival during 12 month follow-up. METHODS: We analyzed data on 489 pts in NYHA II-IV class of HF, referred to our Dept. (mean age was 69 +/- 12). We define doptimal neurohormonal therapy as beta-blocker and ACE-inhibitor in pts with NYHA II, and beta-blocker, ACE-Inhibitor and spironolactone in patients with NYHA III IV class. Pts were divided into groups: group 1--optimal neurohormonal blockade (n = 232, mean age, 67 +/- 11), group 2--non-optimal neurohormonal blockade (n = 257, mean age, 70 +/- 13). Pts were followed for 12 month. RESULTS: Group with optimal therapy were frequent male gender, of ischemic aetiology, and NYHA class II (p < 0.05). Diabetes mellitus, hypertension, left ventricular ejection fraction did not differ the groups (p = NS). Pts with non-optimal therapy were more frequent with prior history of renal dysfunction and anemia at admission (p < 0.05). During 12 month follow-up 12% in optimal vs 40% in non-optimal therapy died (p < 0.005). The rehospitalisation rate during one-year was also significantly higher in pts receiving non-optimal therapy (69% vs 48%, p < 0.005). Cox multivariate analysis showed after adjusting for age, gender, etiology of HF, NYHA functional class, renal dysfunction, EF, had significantly 62% reduction in mortality and 41% reduction in cardiovascular rehospitalisation in pts receiving optimal therapy. CONCLUSIONS: The optimal neurohormonal therapy have favorable effects on outcomes in pts with CHF. This data strongly support that optimalization of care and evidence-based treatment of CHF pts can improve poor prognosis in this group. PMID- 17078491 TI - [Analysis of prognostic factors in patients with autoimmune hepatitis]. AB - Autoimmune hepatitis (AIH) is the chronic inflammatory liver disease of unknown etiology, demonstrating progressive injury of liver, finally leading to insufficiency of this organ. Possible association between clinical forms, course and prognosis of AIH has been considered recently. Aim of this study is to evaluate autoimmune hepatitis exacerbation risk in respect to epidemiological, clinical and laboratory signs demonstrated in patients with freshly diagnosed AIH. Retrospective study included 42 patients hospitalized in Liver Unit of Department of Infectious Diseases Medical University of Bialystok between 1999 and 2003, suspected for autoimmune hepatitis. In majority of patients onset of the disease was sudden and associated with significant increase of ALT activity. The most frequent, first sign of the disease was jaundice, that was observed in over 50% among patients, pruritus was reported by 1/4 patients. In 12 patients (41.4%) beginning of the disease was oligosymptomatic, i.e. without jaundice and pruritus. These patients demonstrated significantly lower ALT activity, bilirubin and gamma-globulin concentrations. Moreover prevalence of antinuclear antibodies in this group was less frequent, and frequency of hospitalization because of AIH exacerbations was also lower. Concluding, oligosynptomatic course of the disease accompanied by lower ALT activity can be related to better prognosis of AIH, and indicate mild course associated with low risk of exacerbations. On the c:her hand faster progression of the disease can be expected in younger patients demonstrating significant increase of ALT activity at the moment of initial diagnosis. PMID- 17078492 TI - [Septic shock in patients with blood diseases. Analysis of clinical situation, treatment and outcome in sixty patients]. AB - Septic shock is one of the major direct causes of death in patients in hematology departments. The aim of the study was to identify situations and factors associated with septic shock in patients with blood diseases. We analyzed the medical treatment of septic shock and its outcome, we based on data from clinical history. MATERIAL AND METHODS: The analyzed group consisted of 60 patients with septic shock, 32 women and 28 men, aged between 20 and 79 years with different blood diseases, hospitalized between 1998 and 2004. RESULTS: Value of APACHE III scale (acute physiology and chronic health evaluation III scale) at the beginning of septic shock was statistically significantly higher for the non-survivors of septic shock than for the survivors (p < 0.001). Septic shock in the analyzed group was associated in similar proportion with presence of Gram-positive (the most often cultured: Staphylococcus epidermidis) and Gram-negative bacteria (the most often cultured: Escherichia coli). The mortality due to septic shock caused by Gram-positive and Gram-negative microorganisms was also similar. We have found no differences in the mortality between neutropenic and non-neutropenic patients. If empirical treatment instituted at the beginning of shock was concordant with result of in vitro sensitivity testing received later, 81% of patients survived septic shock. The survival rate of patients with septic shock receiving discordant antimicrobial treatment was equal 11% and it was statistically less in comparison with patients, when the treatment was concordant (81% vs 11% p < 0.05). When the blood cultures were negative, the adequacy of antibiotic therapy was impossible to confirm and in this group 53% survival rate was noted (81% vs 53%, p < 0.05). In 37 cases, septic shock appeared in patients treated with antibiotics before the onset of septic shock. In this group of patients the highest percent of deaths was noted, when the antibiotic therapy after the moment of starting of septic shock was not changed and was higher in comparison to group, where the treatment was totally changed (64% vs 43% respectively, p < 0.05). CONCLUSIONS: The most important factors influencing survival in septic shock is patient's organ sufficiency and adequacy of applied antibiotics (early goal-directed antibiotic therapy)--in the same way the sensitivity of microorganisms to this treatment. The neutropenia didn't influence an outcome. The best results were achieved in patients whom combinations of three antibiotics were given: beta-lactam + aminoglicoside + glicopeptide antibiotic or carbapenem + glicopeptide antibiotic + fluorochinolon. PMID- 17078493 TI - [A case report of posttraumatic deep vein thrombosis of the lower limbs in a patient with factor V Leiden mutation and the congenital absence of the inferior vena cava]. AB - The congenital absence of the inferior vena cava (AIVC) is a rare vessels' malformation which may predispose to the development of thrombosis. Although AIVC is very rare, its occurrence should be considered in young patients, under 40 years old, with deep vein thrombosis (DVT). We are describing a case of a young male with bilateral deep vein thrombosis, in whom we defined three risk factors for DVT--trauma, factor V Leiden and the absence of the inferior vena cava. It is worth to consider the occurrence of malformation of the inferior vena cava in the young patients with deep vein thrombosis even when the other obvious risk factors are present (trauma, factor V Leiden mutation). The clinical state and the diagnostic process are discussed. PMID- 17078494 TI - [Massive bleeding episodes in patients with acquired haemophilia--case report]. AB - We present two cases with a history of acquired haemophilia with massive haemorrhage in the course of the disease. A 74-year-old man presented to the emergency room with an extensive, progressive swelling and haemorrhage in the region of left knee joint, both upper legs and forearms. Laboratory studies revealed the presence of the factor VIII inhibitor in the titer of 115 Bethesda U/mL, low level of factor VIII activity (19.2%) and severe anemia (Hb - 7.0 g%). The patient was treated with FFP transfusions and prednisone with cyclophosphamide to eradicate factor VIII inhibitor. The remission was achieved and lasts for a two years. A 52-year-old woman presented to the emergency room with an extensive subcutaneous haemorrhage in the region of right knee joint and right lower leg. Laboratory studies revealed the presence of the factor VIII inhibitor in the titer of 30 Bethesda U/mL. The factor VIII activity level was only 4%. The patient did not receive the FFP because the severity of the haemorrhage was low. She was treated with steroids. The factor VIII inhibitor disappeared after 2.5 months of therapy. PMID- 17078495 TI - [Polymyositis and systemic sclerosis symptoms as a clinical manifestation of a chronic graft versus host disease: a case report and literature review]. AB - A case of a patient presenting with polymyositis and systemic sclerosis symptoms due to chronic graft versus host disease is reported. The literature review about similar disease manifestations and hypotheses regarding the pathophysiological origin of the clinical syndromes combined with graft versus host reaction is presented. PMID- 17078497 TI - [Antithyroid drugs and lithium carbonate administration relationship to the results of radioiodine treatment of hyperthyroidism]. PMID- 17078496 TI - [Pneumatosis coli complicated with necrosis and perforation wall of sigmoid colon and diffuse peritonitis]. AB - Pneumatosis coli it is usually benign disease, etiology is still obscure. It can be asymptomatic. Surgery is indicated if are potential complications. Pneumatosis coli characterized by multiple gas-filled cysts present in the submucosa and subserosa. The size of cysta varies between 0.5 and 3 cm. Cysts can occur anywhere along the gastrointestinal tract, but most common are in the small bowel. They occur more frequently in the left than in the right colon. We show a rare case of pneumatosis coli which was compicated of necrosis sigmoid, perforation and diffuse peritonitis. PMID- 17078498 TI - [Disturbances of platelets function in patients with multiple myeloma]. PMID- 17078499 TI - [Influence of ethanol on calcium metabolism]. PMID- 17078500 TI - [Diagnosis and treatment of aspirin-induced asthma]. PMID- 17078501 TI - [Two faces of cytochrome c]. AB - This is an outline of the history of research on cytochrome c. Cytochromes were first discovered by Charles A. MacMunn (1886) and re-discovered by David Keilin (1925) who also identified their function in cell respiration. The role of cytochrome c in the mitochondrial electron transport chain has been well established, thus pointing to a vital role of this haemoprotein in cell function. Yet, towards the end of the last century, a novel role of cytochrome c, namely as a signal molecule for the programmed cell death (apoptosis), has been described. Differences in aminoacid composition of cytochrome c have also been used as markers of biochemical evolution. The article ends with a short biographic note on David Keilin. PMID- 17078502 TI - [The VDAC channel as the mitochondria function regulator]. AB - Regulation of mitochondria physiology, indispensable for proper cell activity, requires an efficient exchange of molecules between mitochondria and cytoplasm at the level of the mitochondrial outer membrane. The common pathway for the metabolite exchange between mitochondria and cytoplasm is the VDAC channel (voltage dependent anion channel), known also as mitochondrial porin. The channel was identified for the first time in 1976 and since that time has been extensively studied. It has been recognized that the VDAC channel plays a crucial role in the regulation of metabolic and energetic functions of mitochondria. In this article we review the VDAC channel relevance to ATP rationing, Ca2+ homeostasis, protection against oxidative stress and apoptosis execution. PMID- 17078503 TI - [Mitochondrial ion channels]. AB - Ion channels are proteins, which facilitate the ions flow throught biological membranes. In recent years the structure as well as the function of the plasma membrane ion channels have been well investigated. The knowledge of intracellular ion channels however is still poor. Up till now, the calcium channel described in endoplasmatic reticulum and mitochondrial porine are the examples of intracellular ion channels, which have been well characterized. The mitochondrial potassium channels: regulated by ATP (mitoK(ATP)) and of big conductance activated by Ca2+ (mitoBK(Ca)), which were described in inner mitochondrial membrane, play a key role in the protection of heart muscle against ischemia. In this review the last date concerning the mitochondrial ion channels as well as they function in cell metabolism have been presented. PMID- 17078504 TI - [Role of mitochondria in reactive oxygen species generation and removal; relevance to signaling and programmed cell death]. AB - Reactive oxygen species (ROS) are universal products of aerobic metabolism, which can be also produced in stress conditions. In eukaryotic cells, mitochondria are the main source of ROS. The main mitochondrial sites of ROS formation are electron carriers of respiratory chain. However, there are also other enzymatic sites capable of ROS generation in different mitochondrial compartments. Reactive oxygen species can cause serious damage to many biological macromolecules, such as proteins, lipids and nucleic acids, which oxidation leads to a lost of their biological properties and eventually to a cell death. Mitochondria, which are also exposed to harmful ROS action, have a defense system that decreases ROS production (first line of defense) or removes generated ROS (second line of defense). Mitochondrial antioxidant system involves proteins that decrease ROS formation, enzymes that directly react with ROS, and non-enzymatic antioxidants that also remove ROS and other oxygen derivatives. Mitochondrial ROS can also act as signal messengers and modify operation of many routes in different cell compartments. Mitochondrial ROS are also important in execution of programmed cell death. PMID- 17078505 TI - [Mechanisms and regulation of the programmed cell death]. AB - The programmed cell death usually is identified with apoptosis, though a scheduled sequence of events can be observed also in autophagy, mitotic catastrophe and, under certain circumstances, in necrosis. Apoptosis begins with activation of the initiator caspases (cysteine proteases) in the signaling complexes: the apoptosome (on the intrinsic or mitochondrial pathway) or the degradosome (on the extrinsic or death receptor pathway). The proteolytic cascade then leads, through activation of downstream caspases and DNases, to digestion of cell components. Mitochondria play a central role in apoptosis by releasing cytochrome c--the essential component of the apoptosome, Smac/Diablo and OmiI/HtrA2--that bind the caspase inhibitors (IAPs), and endonuclease G and AIF- that are responsible for DNA degradation. Those factors get out of mitochondrium through the Bax and Bak protein-containing channels. The process is fast and complete, probably due to mechanoenzyme--driven remodeling of the organellum structure as well as to phospholipid peroxidation and proteolysis in the inner membrane. The release of the mitochondrial factors can be stimulated by protein p53, histone H1.2 and poly(ADP-ribose) that are sent from the nucleus in consequence of a cyto- and genotoxic stress, under the control of cAbl kinase. PMID- 17078506 TI - [Mechanism of peptide bond formation on the ribosome--controversions]. AB - During past five years there have been published many experimental data concerning structure and function of the ribosome. With the presentation of atomic structures we obtained a new data about composition of peptidyl transferase center. It is now obvious that PTC is composed entirely of rRNA. It is also known that the proper substrate alignment is the major factor for ribosome's catalytic activity. However, more detailed mechanism of peptide bond formation on the ribosome still remains unclear. Several issues remain unsolved. For example, are there any chemical components coming from ribosome itself, that enhance the rate of the reaction? Do intact ribosomes perform peptidyltransfer in the same way as the isolated ribosomal subunits that have been the source of most of the data? In this article we present different opinions and controversions around peptide bond formation on the ribosome. PMID- 17078507 TI - [The present-day look at lipid peroxidation]. AB - Lipid peroxidation occurs in physiological conditions and it consists in oxidation of polyunsaturated fatty acids, which are basic biological membrane components. There are two pathway of lipid peroxidation: nonenzymatic and enzymatic. Both of the processes result in generation of various reactive products. Nonenzymatic lipid peroxidation consists in free radical oxidation of polyunsaturated fatty acids while in enzymatic lipid peroxidation the generation of lipid hydroxyperoxides is achieved by insertion of an oxygen molecule at the active centre of lipoxygenase. In this manuscript mechanisms of two processes are described in details. We concentrate on final products of oxidation fatty acids: isoprostanes and neuroprostanes which are regarded as the most important biomarkers of lipid peroxidation as well as on the role of enzymes participating in enzymatic lipid peroxidation process. Moreover we discuss catalytic activity of glutathione peroxidases and transferases--enzymes preventing results of oxidation polyunsaturated fatty acids. PMID- 17078508 TI - [DNA homologous recombination repair in mammalian cells]. AB - DNA double-strand breaks (DSBs) are the most serious DNA damage. Due to a great variety of factors causing DSBs, the efficacy of their repair is crucial for the cell's functioning and prevents DNA fragmentation, chromosomal translocation and deletion. In mammalian cells DSBs can be repaired by non-homologous end joining (NHEJ), homologous recombination (HRR) and single strand annealing (SSA). HRR can be divided into the first and second phase. The first phase is initiated by sensor proteins belonging to the MRN complex, that activate the ATM protein which target HRR proteins to obtain the second response phase--repair. HRR is precise because it utilizes a non-damaged homologous DNA fragment as a template. The key players of HRR in mammalian cells are MRN, RPA, Rad51 and its paralogs, Rad52 and Rad54. PMID- 17078509 TI - [The role of IL-6/sIL-6R complex and its natural inhibitor sgp130 in modulation of inflammatory process]. AB - IL-6/sIL-6R complex belongs to one of the most pleiotropic ligand-receptor system. Participating in leukocytes migration and apoptosis it plays a crucial role in modulation of inflammatory process. Firstly it contributes to resolution of acute inflammatory response regulating a maintaining of organism's homeostasis. Secondly it is a main factor responsible for transition between acute and chronic inflammatory reaction, that underlies the pathogenesis of chronic inflammatory diseases. Presented work focus on the IL-6/sIL-6R structure, function, formation, mechanism of action as well as the role of its natural inhibitor sgp130 in mentioned processes. PMID- 17078510 TI - [Adjuvants--essential components of new generation vaccines]. AB - Adjuvants are essential components of vaccines that augment an immunological reaction of organism. New vaccines based on recombinant proteins and DNA, are more save than traditional vaccines but they are less immunogenic. Therefore, there is an urgent need for the development of new, improved vaccine adjuvants. There are two classes of adjuvants: vaccine delivery systems (e.g. emulsions, microparticles, immune-stimulating complexes ISCOMs, liposomes) and immunostimulatory adjuvants (e.g. lipopolysaccharide, monophosphoryl lipid A, CpG DNA, or muramylpeptides). The discovery of more potent and safer adjuvants may allow to development better prophylactic and therapeutic vaccines against chronic infectious (e.g., HSV, HIV, HCV, HBV, HPV, or Helicobacter pylori) and noninfectious diseases as multiple sclerosis, insulin-dependent diabetes, rheumatoid arthritis, allergy and tumors (e.g., melanoma, breast, or colon cancer). PMID- 17078511 TI - [Subviral molecules of RNA associated with plant ss(+)RNA viruses]. AB - Plant ss(+)RNA viruses besides their genome RNAs often are associated with additional subviral RNA molecules which occur naturally or are generated de novo during infection. There are such molecules like: satellite, defective, defective interfering and chimeric RNAs. Subviral RNAs can not replicate and encapsidate by oneself. Helper viruses supply the protein complexes that are necessary to these processes. The subviral molecules are characterized by small size. Recombination, deletion and accumulation of mutation are the main ways of arising subviral elements, although the origin of satRNAs is unknown. The unique feature of subviral RNAs is their ability to modify of infection progress caused by helper virus. They can attenuate or enhance the intensity of disease symptoms. The overall influence on disease development depends on three-component complex consisting of: plant host-virus' strain--subviral RNA. This article is a synthetic review of information concerning subviral RNA molecules of plant viruses, their structure, functions and origin. PMID- 17078512 TI - [PVD-layering for increased retention of glass fibre reinforced endodontic posts]. AB - For esthetical and biomechanical reasons root canal posts made of fibre reinforced composite (FRC) have gained an important role in clinical application. Additionally, in contrast to metal or ceramic posts, FRC-posts offer the option of removal. Prior to adhesive placement of FRC-posts the root canal dentin of the non vital tooth and the post surface have to be preconditioned. Up to now the post preconditioning has to be proceeded in the chair side technique. This leads to an additional time expense in the clinical treatment schedule. Also a certain risk of errors in application during chair side conditioning procedure is of concern. Modern PVD-technologies can help to make the treatment by the manufacturer well in advance of the clinical use more efficient and reliable, as well as saving clinicians valuable chair-time. For this reason the apical surfaces of the posts were intensively cleaned and activated, PVD-layered and coated by a conserving transparent layer. This coating has the meaning to protect the surface against environmental contamination and allows the try-in of the posts without any risk of damage of the preconditioned surface. To prove the stability of the layer system under simulated clinical conditions pull out tests after 180 days'storage in physiological saline solution have been performed. PMID- 17078513 TI - [In vitro tests of internal tooth whitening agents on colored solutions do not replace tests on teeth]. AB - Two in vitro tests for the assessment of whitening agents were evaluated. A simple, reproducible and cost-effective model using coloured solutions was compared to a tooth model. The following bleaching agents were investigated: 1) Sodium perborate tetrahydrate/30% H2O2, 2) sodium perborate tetrahydrate/3% H2O2, 3) thermocatalytic procedure with 30% H2O2 and 4) Opal-escence Endo. In the model using coloured solutions, the light absorption of the solutions was assessed one and 24 hours after application of the whitening agents. In discoloured teeth, whitening agents were placed in a simulated access cavity. In groups 1, 2, and 4, the agents were left in the cavity for three days, whereas in group 3 the substance was applied three times for 30 seconds. Bleaching processes were repeated three times. All bleaching agents proved effective in the model with coloured solutions, as well as in the tooth model. The effect was most pronounced after the first bleaching step. The results showed, however, that the simplified experiment with coloured solutions cannot replace the tooth model. PMID- 17078514 TI - [Composite resin copings for a perio-overdenture. A newly developed method of making copings for retention with spherical elements]. AB - Today's dentistry should aim at minimizing cost, simplifying techniques, allowing procedural revi sions and finally at facilitating corrections and repairs. The development of direct or indirect light-cured composite resin copings, coupled with ball attachments (Dalbo-Rotex according to Brunner) and a suprastructure which is veneered by means of light-cured composite or acrylic resin, may offer a cost-effective perio-overdenture without compromising on periodontal health and providing optimal esthetics. Since July 2003, eighteen new perio-overdentures with a total of 70 copings made of light-cured microhybrid composite resin were delivered. The aim of this article is to discuss the technical aspects and preliminary results regarding this technique. PMID- 17078515 TI - [Biphosphonate-associated osteonecrosis of the maxilla. Case report and review of the literature]. AB - In recent years, a growing number of reports in the literature have linked osteonecrosis of the jaw bones with intravenously administered bisphosphonates prescribed for the treatment of hypercalcemia of malignancy due to bone lesions of multiple myeloma or bone metastases in patients with breast or prostate cancer. Furthermore, an association between chronic oral bisphosphonate use in patients with osteoporosis or Paget's disease, and bone necrosis in the mandible or maxilla has been demonstrated in numerous case reports and case series in the last couple of years. Therapeutically, osteonecrosis of the jaws seems to be difficult to treat surgically, often resulting in a recurring or even progressing lesion. In the present case report of a bisphosphonate-associated osteonecrosis of the maxilla in a patient with osteoporosis, the current literature will be discussed, and open research questions and potential problems for our daily dental practice routine will be addressed. PMID- 17078516 TI - Doctors in Spanish criminal law: medical criminal responsibility for deaths and injuries caused by negligence in present-day Spain. AB - The doctor who makes a mistake can be taken before a criminal judge in present day Spain. In this paper, I explore some of the articles that refer to doctors' practices in the Spanish Criminal Code, namely article 142.1 (negligent homicide, considered as a major offence), article 152.3 (negligent injury, considered as a less serious offence), as well as the evolution in the views of medical negligence in the last ten years in Spain. Then, I develop a statistical study of three main aspects, namely the number of reports and lawsuits that have been recorded, the number of criminal proceedings that have been undertaken, and the number of judgments that have acquitted or convicted a doctor. This leads to an analysis of the consequences of the application of the system, as well as its effects on insurance policies. In the last section of my paper, I offer some conclusions. PMID- 17078517 TI - Consent for organ and tissue retention in British law in the light of the Human Tissue Act 2004. AB - The experience from the scandals in hospitals at Liverpool and Bristol in the UK where retention of tissue and organs was undertaken without the consent of the parents and relatives raised serious concerns regarding the efficacy of the existing Human Tissue Act 1961, in England and the operation of the law by medical practitioners. In the aftermath of these damaging scandals a combination of public distrust and government overreaction has led to the enactment of new legislation, the Human Tissue Act 2004, which is aiming to prevent any further instances of the retention of organs and tissue from dead children or adults without their next of kin's consent or knowledge. However, scientists have expressed concerns that such changes might seriously endanger several medical research programmes, and also tissue and organ donation for transplantation. The aim of this article is to highlight important issues raised by existing practice in the post-mortem examinations in the UK and the lessons learnt from this and to discuss the benefits and the potential problems arising from the new Act. PMID- 17078518 TI - Compensation of asbestos victims in France. AB - The alarming development of pathologies linked to asbestos led to the creation in France of two funds to indemnify the victims of asbestos-related illnesses: the FCAATA (Fund for asbestos workers who take early retirement), which compensates for their reduced life expectancy, and the FIVA (Indemnification fund for asbestos victims) which ensures full compensation for harm suffered by asbestos victims. PMID- 17078519 TI - Alternative, complementary and traditional medicine in Malaysia. AB - This paper sets out the practice of traditional, alternative and/or complementary medicine in Malaysia. It gives an overview of the types of alternative medicine available, and the legal regulation, or lack of it within the current setting. The relevant policies and governmental action in this area are highlighted. Relevant case law decisions in this area are also included. The practice of spiritual healing as one form of traditional medicine, and its role within the spectrum of alternative medicine is dealt with briefly. The significant question of integration of alternative medicine within the existing allopathic system is addressed. The paper concludes that as interest in, and usage of alternative medicine is not likely to decrease, certain measures must be taken by the relevant authorities to ensure among others, the safety and efficacy of these medicines. PMID- 17078520 TI - Comparisons of the interactions of health care delivery and medico-legal practice between Australia and Singapore. AB - Australia and Singapore have similar standards of health care. The one major difference in the two health care systems is the cost to the patient at the point of care. The Medicare system in Australia provides partial to complete subsidy for health care delivery in the public hospitals. In Singapore, the patient has to bear the cost of their health care when needed, with some government subsidies. Studies in the variations between two health care systems, where the costs to the government and individuals are clearly dissimilar, but the health outcomes are similar, can be educational for health law specialists. The methods in which patients obtain recompense for their grievances can help both countries understand how to determine and improve standards of health care communication. Having worked in both systems, the relative values of each and their effects on medical litigation will be discussed. PMID- 17078521 TI - The tension between cost-utility analysis and preventive public health. AB - Public health programs must often work under severe budgetary constraints. It would appear then that cost-utility analysis could offer decision-makers a practical approach to prioritizing public health interventions. Unfortunately, cost-utility analysis is a poor tool for programmatic decision-making in public health because it systematically downgrades interventions aimed at prevention, and because it is not designed to help guide multi-dimensional, multi-input community public health programs. Thus, a different kind of prioritization framework is needed, one that is better aligned with community action and the preventive aims of public health. PMID- 17078522 TI - Biotechnology entrepreneurship and ethics: principles, paradigms, and products. AB - Biotechnology, whether in the context of new drugs derived from DNA and genetic technology, genetically modified food, or biologics making use of living cells, raises ethical concerns at a variety of different levels. At the research level, there is concern that the very nature of research is being subverted, rather than enhanced, by entrepreneurship. This area of ethical concern has intensified in the United States as a result of the conflicts of interests resulting from the growing alliance between University academia and private industry in the research enterprise. As we travel down the research path into development of a drug or technology, ethical questions arise with respect to protecting human subjects and society from danger and exploitation by researchers. As development gives way to marketing and dissemination of a new product, government regulators are pressed to get drugs and biologics through the regulatory pipeline into the market faster, walking an ethical tightrope between speed and safety. As new biotechnology products enter the market place, doctors and patients traverse yet another tightrope, that between unknown risk and the promise of benefit. And finally, patent protection is increasingly viewed as a unethical culprit in keeping prices high and depriving the global poor from lifesaving drugs and biologics. Bioethics has, to date, been largely a creation of Western research and medicine. As such it is wholly inadequate to respond to the cascade of ethical issues that flow from a vibrant biotechnology industry. And if biotechnology is in its infancy, as most believe, it is crucial that scientists, entrepreneurs and governments engage in dialogue about the ethical and societal questions raised on the road of scientific progress. PMID- 17078523 TI - Ethics in clinical drug trial research in private practice. AB - INTRODUCTION: Private clinics and clinicians have been involved in clinical drug trials for approximately two decades. This paper reviews the ethical consideration inherent in this process. METHODS: Involvement of a single community based, private, Australian neurological clinic in the conduct of trials was audited. Changes in ethical considerations were analysed. RESULTS: The clinic previously audited its clinical trial involvement, starting with pharmaceutical company orchestrated trials. These were vetted by hospital based ethics committees (ECs) which then refused to review private research. A private EC accommodating NH & MRC standards was formed to assess private research. Indemnity concerns forced return to institutional ECs with government guaranteed indemnification. Trials evolved to investigator initiated, company sponsored studies thence a company asking the clinic to devise, sponsor and manage a trial. The latter relegated trial co-ordination to the clinic which would control publication thereby creating new ethical standards. DISCUSSION: Private practice trial involvement evolved from reluctant inclusion to a pivotal role in privately sponsored studies. Access to ECs is government endorsed and publication is independent for investigator-sponsored trials. There has been modification of standard operating procedures and enhanced ethical standards. PMID- 17078524 TI - Ethical misconduct by abuse of conscientious objection laws. AB - This paper addresses laws and practices urged by conservative religious organizations that invoke conscientious objection in order to deny patients access to lawful procedures. Many are reproductive health services, such as contraception, sterilization and abortion, on which women's health depends. Religious institutions that historically served a mission to provide healthcare are now perverting this commitment in order to deny care. Physicians who followed their calling honourably in a spirit of self-sacrifice are being urged to sacrifice patients' interests to promote their own, compromising their professional ethics by conflict of interest. The shield tolerant societies allowed to protect religious conscience is abused by religiously-influenced agencies that beat it into a sword to compel patients, particularly women, to comply with religious values they do not share. This is unethical unless accompanied by objectors' duty of referral to non-objecting practitioners, and governmental responsibility to ensure supply of and patients' access to such practitioners. PMID- 17078525 TI - Milestones in the evolution of standards for experimental treatment or research. AB - The abuses in experimentation that marked the 20th century has resulted in regulations. Standards for experimental treatment or for research involving human subjects has been a major development of the twentieth century, coming about in response to the horrendous experiments carried out by Nazi Germany and also in the United States and elsewhere. How these regulations have fared is discussed herein. PMID- 17078526 TI - Collection of fetal blood for stem cell research and therapy. AB - Stem cell research has generated novel therapeutic opportunities at the expense of new ethical and legal problems. Its promoters recommended early clamping of the umbilical cord to maximize the amount of acquired fetal blood. Fear has been expressed, therefore, that the donor could be compromised by this approach. Actually, the problem is more complex than generally assumed. In certain clinical situations the neonate may benefit from or become harmed by additional blood volume. Gravity influences the direction of umbilical blood flow and, thus the consequences of early or delayed cord clamping. Therefore, vaginal birth promotes blood flow from the placenta to the fetus, whereas delivery by cesarean section usually has the opposite effect. Largely ignored in the course of the relevant debates, the above facts require consideration. The controversy may be beneficial in the long run by drawing attention to this relatively neglected aspect of perinatal medicine. PMID- 17078527 TI - M@TIC. Medicine Assisted by Information and Communication Technology: conflicts, responsibility and liability. AB - The term M@TIC, is that which encompasses all Medicine Assisted by Technology on Information and Communication. The development of these techniques brings up many ethical and legal conflicts, mainly because medical science has developed much faster than the law. Justice cannot be properly served if we do not have the regulations to help mankind to avoid aberrant behavior by medical practitioners in this field. M@TIC is still regarded as an experimental research procedure. Not all that is technically feasible is ethically acceptable. There are many potential risks associated with M@TIC and it is largely considered, and so it is understood by doctors, that any damage to the patient would be the fault of the system, never the responsibility of the doctor. It frightens one to think that this fact may be used, in the future, as a shield to protect negligent medical doctors from malpractice suits. PMID- 17078528 TI - Privacy and confidentiality--old concept, new challenges. AB - This paper concerns a privacy and confidentiality problem in cases of exceptional situations. It is primarily aimed at using DNA samples, and at a breach of genetic data protection. European legal provisions are especially mentioned. PMID- 17078529 TI - Light and electron microscopic studies on liver histology in chicks fed aflatoxin. AB - This study was carried out under both light and electron microscopy to investigate the effects on liver carbohydrate and lipid metabolism caused by aflatoxin (AF) fed to chicks. Twenty broiler chicks were used. The birds were housed in electrically heated battery cages and exposed to light for 24 h. Feed and water were provided ad libitum. The animals were allocated to two groups each made up of 10 broilers. Total aflatoxin levels of zero (0) and 5 mg of AF/kg feed (81.05% AFB1, 8.79% AFG1, 6.06% AFB2, and 4.10% AFG2) added to a commercial diet, were fed to chicks from hatching up to 3 weeks of age, when the experiment was terminated. The chicks were executed by cervical dislocation and liver samples were obtained for light and electron microscopy. Oil red 'O', Sudan Black B, periodic acid Schiff (PAS) and Best's carmine stains were used to reveal fat and glycogen in the liver. Histological changes in hepatocytes included increased lipid droplets, high glycogen content, and mild mononuclear cell infiltration in the portal areas. Ultrastructural findings were destruction of rough endoplasmic reticulum (rER), reduction in mitochondrial size, enlargement of bile canaliculi, and cisternal dilatation of the smooth endoplasmic reticulum (sER). PMID- 17078530 TI - [Investigations on the influence of selected compulsory measures on clinically relevant haematological and blood-chemical parameters of racing pigeons (Columba livia f. dom.)]. AB - In the presented study the influence of stress and environmental factors on selected haematological and blood-chemical parameters in racing pigeons was examined. Blood was taken at three defined days and haematological as well as blood-chemical parameters of clinical relevance were determined. In comparison to reference values published for pigeons, the majority of the values obtained in this study were within physiological borders. The daily handling of the pigeons did not have any significant effect on the examined parameters. Also the heterophile/lymphocyte ratio did not show any changes characteristic of a stress reaction. In contrast, after change of the housing dies with pigeons originating normally from a flock, the first blood sampling should be performed after a 4 or 5-day lasting period of acclimatisation to the the individual housing conditions. PMID- 17078531 TI - Changes in haematological profile of common pheasant (Phasianus colchicus) induced by transit to pheasantry. AB - The aim of this study was to assess haematological changes in hand-reared pheasants (Phasianus colchicus) transported from intensive housing facilities to a pheasantry. Selected haematological parameters were monitored in a group of 100 pheasants (50 males and 50 females) aged of 9 weeks that were transported for 4 hours by a covered lorry in crates, with a total body weight of 12 +/- 0.5 kg per crate (Group C12 - floor space: 290 cm2/kg) and with a total body weight of 18 +/ 0.5 kg per crate (Group C18 - floor space: 195 cm2/kg). Blood samples were taken from 10 randomly selected males and 10 females before transport (CON group) and 20 hours after transport (C12 and C18 groups). Examinations consisted in determining the total erythrocyte and leukocyte counts, haematocrit values, haemoglobin levels and differential leukocyte counts, whereby the proportions of heterophil, basophil and eosinophil granulocytes, lymphocytes and monocytes of the total leukocytes were computed. The changes in the parameters of red blood cell count were manifested by an increase (P < 0.01) in the haemoglobin level, MCH (mean cell haemoglobin) and MCHC (mean cell haemoglobin concentration) values and a decrease (P < 0.01) in the total erythrocyte count and haematocrit level in both C12 and C18 pheasants, when compared with the control group of non transported pheasants. C18 pheasants exhibited also a significant increase (P < 0.05) in MCV (mean cell value) value. When analyzing differential leukocyte counts, C18 pheasants showed a decrease (P < 0.01) in heterophil counts and H/L ratio, whereas values in C12 pheasants did not differ from the non-transported control group. Individual counts of lymphocytes were decreased (P < 0.05) in C12 pheasants, whereas basophil counts were increased (P < 0.01) in both C12 and C18 pheasants. Total leukocyte count was decreased (P < 0.01) in C12 and C18 pheasants. In conclusion, the specific requirements of pheasants, as primarily wild animals, for the density in crates should be respected during transportation and they should be transported at lower densities than other poultry species, at least 290 cm2/kg live weight should be provided. PMID- 17078532 TI - [Effects of non-starch-polysaccharides in nutrition of monogastric animals]. AB - Polysaccharides are widespread biopolymers, which quantitatively represent the most important group of nutrients in botanical feed. In animal nutrition polysaccharides, which cannot be degraded by endogeneous enzymes and therfore reach the colon almost indigested, are summarized as "non-starch-polysaccharides (NSP)". Individual NSP groups have different chemical and physical characteristics, that result in various effects on physiology of intestine and on organism in general. They affect e.g. digesta passage, nutrient resorption, microbial growth and activity, lipid metabolism and immune systeme. The physiological effects lead to conclusion that some polysaccharides are suitable tools in health management of farm animals. However, limitations result from antinutritive effects and from little scientific knowledge about modes of action in vivo. PMID- 17078533 TI - [Animal nutrition for veterinarians--case study: colic in ponies in a "petting zoo" caused by ingestion of higher amounts of wheat (Triticum aestivum L.)]. AB - Nutritional disorders in domestic or pet animals depend not seldom on special situations, for example when different influences coincide concerning keeping, housing and offering of feed or water. These lead suddenly to high-risk situations, that can cause sometimes the death of the animal. The following case report deals with two ponies, that were kept on a so-called "petting zoo" and showed repeatedly colic symptoms (always after a weekend). During surgery in the stomach light yellow balls were found, that can be explained by ingestion of 1.0 to 1.5 kg wheat. This cereals consist--depending on variety--gluten, that can cause the observed conglobates. Due to the forming of such balls that can lead to health disorders in form of colics, gastritis or ruptures of the stomach the feeding of high amounts of wheat should be avoided. Snacks rich in fiber or high amounts of roughage (straw, hay) that are fed before opening of the zoo could be a practical alternative. PMID- 17078534 TI - [Immunosuppressants for genetic disorders--drug treatment of choice]. PMID- 17078535 TI - Landfill leachate production, quality and recirculation treatment in northeast China. AB - Landfill leachate recirculation treatment process is a promising and economic way in northeast China due to the distinct season variation. In order to study the impact of recirculation on leachate production and its quality, two simulated sanitary landfill reactors, one with leachate recirculation and the other without, were placed outdoor in Harbin. The two-year experimental results indicated that leachate production was reduced by 86% with leachate recirculation and less or no facility for leachate treatment required. Recirculation could improve the leachate quality dramatically with lower COD, SS, NH4(+)-N and higher pH level. Recirculation also could enhance waste degradation and stabilization and improve the efficiency of landfill. This study confirmed that leachate recirculation was a very feasible way for on-site landfill leachate treatment in cold areas. PMID- 17078536 TI - Treatment of phosphate-containing oily wastewater by coagulation and microfiltration. AB - The oily wastewater generated from pretreatment unit of electrocoating industry contains oils, phosphate, organic solvents, and surfactants. In order to improve the removal efficiencies of phosphate and oils, to mitigate the membrane fouling, coagulation for ceramic membrane microfiltration of oily wastewater was performed. The results of filtration tests show that the membrane fouling decreased and the permeate flux and quality increased with coagulation as pretreatment. At the coagulant Ca (OH)2 dosage of 900 mg/L, the removal efficiency of phosphate was increased from 46.4% without coagulation to 99.6%; the removal of COD and oils were 97.0% and 99.8%, respectively. And the permeate flux was about 70% greater than that when Ca(OH)2 was not used. The permeate obtained from coagulation and microfiltration can be reused as make-up water, and the recommended operation conditions for pilot and industrial application are transmembrane pressure of 0.10 MPa and cross-flow velocity of 5 m/s. The comparison results show that 0.2 microm ZrO2 microfilter with coagulation could be used to perform the filtration rather than conventional ultrafilter, with very substantial gain in flux and removal efficiency of phosphate. PMID- 17078537 TI - Suspended particle effects on ClO2/ultraviolet light combined disinfection of effluent. AB - The concentration of suspended solids of effluent often varies in a wide range, therefore the dose of ultraviolet light (UV) in disinfection process needs to be adjusted to meet the disinfection criterion at a high frequency, and the desired disinfection effect is difficult to be ensured. The particles size and particle associated fecal coliform (F.C.) contribution, and their influence on UV disinfection were investigated when ClO2 and UV combined disinfection process was used. The results showed that suspended solids content had a major impact on UV disinfection efficiency, especially the large particle size fraction. Particles (D>10 microm) associated F. C. were difficult to be disinfected and were the main part of the tailings of F.C. inactivation curve. Pre-ClO2 oxidation could reduce the number of particles in effluent, and make large particles decrease to small ones. Therefore, the influence of particles on UV disinfection could be reduced after pre-ClO2 oxidation, and the resistance ability to particle loadings of combined process was enhanced. Moreover, the combined process has a lot of advantages, such as low toxicity, low operational/maintenance costs; it is also convenient to be established in the existing wastewater plant or the new planned one. PMID- 17078538 TI - Biodegradation of high concentration phenol containing heavy metal ions by functional biofilm in bioelectro-reactor. AB - Functional microorganisms to high concentration phenol containing Cr6+ and Pb2+ were cultured and biofilm was formed on polypropylene packings in bioelectro reactor. It was found that the biodegradation capability of such biofilm to phenol changed with the applied voltage. Under the optimal electric field conditions (voltage of 3.0 V, electric field of strength 17.7 V/m and current density of 1.98 A/m2), biodegradation efficiency of phenol aof concentration of 1200 mg/L increased 33% compared to the instance without applying electric field. However, voltage had inverse effect on biodegradation, as microorganisms were killed under strong electric field. Voltage had little effect on heavy ions elimination. Higher absorption rate of Cr6+ and Pb2+ was observed when changing pH from acidic to neutral. The experiment results indicated that, after treatment, 10 L phenol of 2400 mg/L was biodegraded completely within 55 h and concentrations of Cr6+ and Pb2+ dropped to less than 1 mg/L within 12 h and 6 h, from initial values of 50 mg/L and 30 mg/L, respectively. PMID- 17078539 TI - Supercritical gasification for the treatment of o-cresol wastewater. AB - The supercritical water gasification of phenolic wastewater without oxidant was performed to degrade pollutants and produce hydrogen-enriched gases. The simulated o-cresol wastewater was gasified at 440-650 degrees C and 27.6 MPa in a continuous Inconel 625 reactor with the residence time of 0.42-1.25 min. The influence of the reaction temperature, residence time, pressure, catalyst, oxidant and the pollutant concentration on the gasification efficiency was investigated. Higher temperature and longer residence time enhanced the o-cresol gasification. The TOC removal rate and hydrogen gasification rate were 90.6% and 194.6%, respectively, at the temperature of 650 degrees C and the residence time of 0.83 min. The product gas was mainly composed of H2, CO2, CH4 and CO, among which the total molar percentage of H2 and CH4 was higher than 50%. The gasification efficiency decreased with the pollutant concentration increasing. Both the catalyst and oxidant could accelerate the hydrocarbon gasification at a lower reaction temperature, in which the catalyst promoted H2 production and the oxidant enhanced CO2 generation. The intermediates of liquid effluents were analyzed and phenol was found to be the main composition. The results indicate that the supercritical gasification is a promising way for the treatment of hazardous organic wastewater. PMID- 17078540 TI - Effects of UV intensity and water turbidity on microbial indicator inactivation. AB - The effects of UV intensity and turbidity on selected microbial indicator inactivation were investigated. Results showed that UV disinfection was effective in killing all the selected microbial indicators, the resistance order of the microorganisms was as follows: MS-2 coliphage > Bacillus subtilis > E. coli > Staphylococcus aureus and Candida albicans. UV intensity had influence on the inactivation of all the microorganisms, high UV disinfection efficency was obtained with higher UV intensity. Turbidity had impact on the bacteria inactivation rate, but there was no evidence that turbidity had any negative contribution to MS-2 coliphage. Under the same UV dosage, higher UV intensity could overcome the negative influence of turbidity on UV performance, enhanced microorganism inactivation effect in turbidity water. PMID- 17078541 TI - Removal of organic matter and nitrogen from distillery wastewater by a combination of methane fermentation and denitrification/nitrification processes. AB - The distillery wastewater of Guangdong Jiujiang Distillery, which is characteristic of containing high organic matters and rich total nitrogen, was treated by a combination of methane fermentation and denitrification/nitrification processes. 80% of COD in the raw wastewater was removed by methane fermentation at the COD volumetric loading rate of 20 kg COD/(m3 x d) using the expanded granule sludge bed (EGSB) process. However, almost all the organic nitrogen in the raw wastewater was converted into ammonia by ammonification there. Ammonia and volatile fatty acids (VFA) remaining in the anaerobically treated wastewater were simultaneously removed utilizing VFA as an electron donor by denitrification occurring in the other EGSB reactor and nitrification using PEG-immobilized nitrifying bacteria with recirculation process. An aerobic biological contact oxidization reactor was designed between denitrification/nitrification reactor for further COD removal. With the above treatment system, 18000-28000 mg/L of COD in raw wastewater was reduced to less than 100 mg/L. Also, ammonia in the effluent of the system was not detected and the system had a high removal rate for 900-1200 mg/L of TN in the raw wastewater, only leaving 400 mg/L of nitrate nitrogen. PMID- 17078542 TI - Kinetics study on photochemical oxidation of polyacrylamide by ozone combined with hydrogen peroxide and ultraviolet radiation. AB - An investigation on the process of ozone combined with hydrogen peroxide and ultraviolet radiation has been carried out in order to establish the kinetics for photochemical oxidation of polyacrylamide (PAM) in aqueous solution. Effects of operating parameters, including initial PAM concentration, dosages of ozone and hydrogen peroxide, UV radiation and pH value on the photochemical oxidation of PAM, have been studied. There was an increase in photochemical oxidation rate of PAM with increasing of dosages of O3, H2O2 and ultraviolet radiation. Upon increasing of the initial PAM concentration, the photochemical oxidation rate of PAM decreased. Slight effect of pH value on the photochemical oxidation rate of PAM was observed in the experiments. The kinetics equation for the photochemical oxidation of PAM by the system has been established. PMID- 17078543 TI - Performance of hybrid constructed wetland systems for treating septic tank effluent. AB - The integrated wetland systems were constructed by combining horizontal-flow and vertical-flow bed, and their purification efficiencies for septic tank effluent were detected when the hydraulic retention time (HRT) was 1 d, 3 d, 5 d under different seasons. The results showed that the removal efficiencies of the organics, phosphorus were steady in the hybrid systems, but the removal efficiency of total nitrogen was not steady due to high total nitrogen concentration in the septic tank effluent. The average removal rates of COD (chemical oxygen demand) were 89%, 87%, 83%, and 86% in summer, autumn, winter and spring, respectively, and it was up to 88%, 85%, 73%, and 74% for BOD5 (5 d biochemical oxygen demand) removal rate in four seasons. The average removal rates of TP (total phosphorous) could reach up to 97%, 98%, 95%, 98% in four seasons, but the removal rate of TN (total nitrogen) was very low. The results of this study also indicated that the capability of purification was the worst in winter. Cultivating with plants could improve the treated effluent quality from the hybrid systems. The results of the operation of the horizontal-flow and vertical-flow cells (hybrid systems) showed that the removal efficiencies of the organics, TP and TN in horizontal-flow and vertical-flow cells were improved significantly with the extension of HRT under the same season. The removal rate of 3 d HRT was obviously higher than that of 1 d HRT, and the removal rate of 5 d HRT was better than that of 3 d HRT, but the removal efficiency was not very obvious with the increment of HRT. Therefore, 3 d HRT might be recommended in the actual operation of the hybrid systems for economic and technical reasons. PMID- 17078544 TI - Role of extracellular exopolymers on biological phosphorus removal. AB - Three sequencing batch reactors supplied with different carbon sources were investigated. The system supplied with glucose gained the best enhanced biological phosphorus removal although all of the three reactors were seeded from the same sludge. With the measurement of poly-beta-hydroxyalkanoate (PHA) concentration, phosphorus content in sludge and extracellular exopolymers (EPS) with scanning electron microscopy (SEM) combined with energy dispersive spectrometry (EDS), it was found that the biosorption effect of EPS played an important role in phosphorus removal and that the amount of PHA at the end of anaerobic phase was not the only key factor to determine the following phosphorus removal efficiency. PMID- 17078545 TI - Estimating the change of porosity in the saturated zone during air sparging. AB - Air sparging is a remedial method for groundwater. The remedial region is similar to the air flow region in the saturated zone. If soil particles are transported during air sparging, the porosity distributions in the saturated zone change, which may alter the flow path of the air. To understand better the particle movement, this study performed a sandbox test to estimate the soil porosity change during air sparging. A clear fracture was formed and the phenomenon of particle movement was observed when the air injection was started. The moved sand filled the porous around the fracture and the reparked sand filled the fracture, reducing the porosity around the fracture. The results obtained from the photographs of the sandbox, the current measurements and the direct sand sample measurements were close to each other and are credible. Therefore, air injection during air sparging causes sand particle movement of sand, altering the characteristic of the sand matrix and the air distribution. PMID- 17078546 TI - Spatial and temporal variations of water quality in Cao-E River of eastern China. AB - Evaluation and analysis of water quality variations were performed with integrated consideration of water quality parameters, hydrological-meteorologic and anthropogenic factors in Cao-E River, Zhejiang Province of China. Cao-E River system has been polluted and the water quality of some reaches are inferior to Grade V according to National Surface Water Quality Standard of China (GB2002). However, mainly polluted indices of each tributary and mainstream are different. Total nitrogen (TN) and total phosphorus (TP) in the water are the main polluted indices for mainstream that varies from 1.52 to 45.85 mg/L and 0.02 to 4.02 mg/L, respectively. TN is the main polluted indices for Sub-watershed I, II, IV and V (0.76 to 18.27 mg/L). BOD5 (0.36 to 289.5 mg/L), CODMn (0.47 to 78.86 mg/L), TN (0.74 to 31.09 mg/L) and TP (0 to 3.75 mg/L) are the main polluted indices for Sub-watershed III. There are tow pollution types along the river including nonpoint source pollution and point source pollution types. Remarkably temporal variations with a few spatial variations occur in nonpoint pollution type reaches (including mainstream, Sub-watershed I and II) that mainly drained by arable field and/or dispersive rural dwelling district, and the maximum pollutant concentration appears in flooding seasons. It implied that the runoff increases the pollutant concentration of the water in the nonpoint pollution type reaches. On the other hand, remarkably spatial variations occur in the point pollution type reaches (include Sub-watershed III, IV and V) and the maximum pollutant concentration appears in urban reaches. The runoff always decreases the pollutant concentration of the river water in the seriously polluted reaches that drained by industrial point sewage. But for the point pollution reaches resulted from centralized town domestic sewage pipeline and from frequent shipping and digging sands, rainfall always increased the concentration of pollutant (TN) in the river water too. Pollution controls were respectively suggested for these tow types according to different pollution causes. PMID- 17078547 TI - Surface-exchange of NOx and NH3 above a winter wheat field in the Yangtze Delta, China. AB - A four-dynamic-chamber system was constructed to measure NOx and NH3 surface exchange between a typical wheat field and the atmosphere in the Yangtze Delta, China. The average fluxes ofNO, NO2 and NH3 were 79, -5.6 and -5.1 ngN/(m2 x s), and 91, -1.8 and 23 ngN/(m2 x s), respectively for the wheat field and the bare soil. The NO flux was positively correlated with soil temperature and the fluxes of NO2 and NH3 were negatively correlated with their ambient concentrations during the investigated period. The compensation point of NO2 between the wheat field and the atmosphere was 11.9 microg/m3. The emissions of NO-N and NH3-N from the urea applied to the wheat field were 2.3% and 0.2%, respectively, which indicated that the main pathway of N loss from the investigated winter wheat field was NO. Application of a mixture of urea and lignin increased the emissions of NO, but also greatly increased the yield of the winter wheat. PMID- 17078548 TI - Chemical characteristics of aerosol particles (PM2.5) at a site of Horqin Sand land in northeast China. AB - The objective of this study was to characterize the mass concentration and chemical composition of aerosol particles (PM2.5) collected at Tongliao (Inner Mongolia Autonomous Region, China), a site in Horqin Sand-land in northeast China. During spring 2005, the mass concentration for PM2.5 was (126 +/- 71)microg/m3 in average. Five dust storm events were monitored with higher concentration of (255 +/- 77)microg/m3 in average than the non dusty days of (106 +/- 44)microg/m3. Concentrations for 20 elements were obtained by the PIXE method. Mass concentrations of Al, Mg, Si, K, Ca, Ti, Mn, and V, which increased with the PM2.5 concentration, were higher than the pollution elements (S, Cl, Zn, Ar, Se, Br, and Pb). Enrichment factor relative to crust material was also calculated, which showed dust trace elements were mainly from earth upper crust and pollution elements were dominated the anthropogenic aerosols. The Si/Al, Ca/Al, and Fe/Al ratios in PM2.5 samples at Tongliao were 4.07, 0.94, and 0.82, respectively, which were remarkably different with those on other source regions, such as "Western desert source region", "North desert source region" and central Asia source. Air mass back-trajectory analysis identified three kinds of general pathways were associated with the aerosol particle transport to Tongliao, but have the similar elemental ratios, implying that elemental signatures for dust aerosol from Horqin Sand-land were different with other regions. PMID- 17078549 TI - Temporal variations of CO2 concentration near land surface and its response to meteorological variables in Heihe River Basin, northwest China. AB - Atmospheric CO2 concentration (CC) near land surface and meteorological variables have been measured at four sites, named Yeniugou (alpine meadow and permafrost), Xishui (mountainous forest), Linze (oasis edge) and Ejina (lower desert), respectively, in Heihe River Basin, northwest China. The results showed that, the half hourly CC at night was larger than in daytime, and the daily averaged CC was the largest in winter. The averaged CC of 932 d at the Linze was about 418 ppm, was about 366 ppm in the 762 d at the Ejina. In the same period from September 23 to November 9, 2004, the averaged CC was about 625, 334, 436 and 353 ppm, at Yeniugou, Xishui, Linze and Ejina, respectively. The linear relationship between daily averaged CC and air temperature T was negative, between CC and relative humidity (RH) was positive. The linear CC-atmospheric pressure (AP) relationship was negative at the Linze and Yeniugou, was positive at the Ejina. The relationship between CC and global radiation R was exponent, and soil temperature Ts was negative linear, and soil water content was complex. The correlation between CC and wind speed was not existent. Using meteorological variables together to simulate CC, could give good results. PMID- 17078550 TI - Numerical simulation of PAHs sorption/desorption on soil with the influence of Tween80. AB - In this paper, the influences of inionic surfactant Tween80 on polycyclic aromatic hydrocarbons (PAHs) sorption/desorption on artificially contaminated soil were studied, and gamma model was applied to simulate the influences. Results showed that, with the use of Tween 80, the sorption behaviors of PAHs on soil altered significantly. Adsorbed Tween 80 increased the sorption amount of PAHs while the dissolved Tween80 increased the apparent solubility of PAHs. These two processes exert influences on the sorption coefficient of PAHs in soil-water system, which can be depicted by apparent sorption coefficient. The partition coefficients (the soil/water partition coefficient of PAHs and surfactants obtained from sorption experiments) and statistical parameters used in the amended gamma model were obtained in independent experiments. With these parameters, the gamma model could provide a satisfactory independent prediction of PAHs release from soil to aqueous phase at two surfactant concentrations. PMID- 17078551 TI - Effects of Glomus mosseae on the toxicity of heavy metals to Vicia faba. AB - A glasshouse pot experiment was conducted to investigate effects of the arbuscular mycorrhizal fungus Glomus mosseae on the growth of Vicia faba and toxicity induced by heavy metals (HMs) (Cu, Zn, Pb and Cd) in a field soil contaminated by a mixture of these metals. There was also uninoculation treatment (NM) simultaneously. Mycorrhizal (GM) plants hav e significantlyincreased growth and tolerance to toxicity induced by heavy metals compared with NM plants. P uptake was significantly increased in GM plants. Mycorrhizal symbiosis reduced the transportation of HMs from root to shoot by immobilizing HMs in the mycorrhizal, shown by increasing the ratios of HMs from root to shoot. Oxidative stress, which can induce DNA damage, is an important mechanism of heavy metal toxicity. GM treatment decreased oxidative stress by intricating antioxidative systems such as peroxidases and non-enzymic systems including soluble protein. The DNA damage induced by heavy metals was detected using comet assay, which showed DNA damage in the plants was decreased by the GM treatment. PMID- 17078552 TI - Removal of heavy metals from a contaminated soil using tartaric acid. AB - This study reports the feasibility of remediation of a heavy metal (HM) contaminated soil using tartaric acid, an environmentally-friendly extractant. Batch experiments were performed to test the factors influencing remediation of the HM contaminated soil. An empirical model was employed to describe the kinetics of HM dissolution/desorption and to predict equilibrium concentrations of HMs in soil leachate. The changes of HMs in different fractions before and after tartaric acid treatment were also investigated. Tartaric acid solution containing HMs was regenerated by chestnut shells. Results show that utilization of tartaric acid was effective for removal of HMs from the contaminated soil, attaining 50%-60% of Cd, 40%-50% of Pb, 40 %-50% of Cu and 20%-30% of Zn in the pH range of 3.5-4.0 within 24 h. Mass transfer coefficients for cadmium (Cd) and lead (Pb) were much higher than those for copper (Cu) and zinc (Zn). Sequential fractionations of treated and untreated soil samples showed that tartaric acid was effective in removing the exchangeable, carbonate fractions of Cd, Zn and Cu from the contaminated soil. The contents of Pb and Cu in Fe-Mn oxide fraciton were also significantly decreased by tartaric acid treatment. One hundred milliliters of tartaric acid solution containing HMs could be regenerated by 10 g chestnut shells in a batch reactor. Such a remediation procedure indicated that tartaric acid is a promising agent for remediation of HM contaminated soils. However, further research is needed before the method can be practically used for in situ remediation of contaminated sites. PMID- 17078553 TI - Field released transgenic papaya effect on soil microbial communities and enzyme activities. AB - Soil properties, microbial communities and enzyme activities were studied in soil amended with replicase (RP)-transgenic or non-transgenic papaya under field conditions. Compared with non-transgenic papaya, significant differences (P<0.05) were observed in total nitrogen in soils grown with transgenic papaya. There were also significant differences (P<0.05) in the total number of colony forming units (CFUs) of bacteria, actinomycetes and fungi between soils amended with RP transgenic plants and non-transgenic plants. Compared with non-transgenic papaya, the total CFUs of bacteria, actinomycetes and fungi in soil with transgenic papaya increased by 0.43-1.1, 0.21-0.80 and 0.46-0.73 times respectively. Significantly higher (P<0.05) CFUs of bacteria, actinomycetes and fungi resistant to kanamycin (Km) were obtained in soils with RP-transgenic papaya than those with non-transgenic papaya in all concentrations of Km. Higher resistance quotients for Kmr (kanamycin resistant) bacteria, actinomycetes and fungi were found in soil planted with RP-transgenic papaya, and the resistance quotients for Kmr bacteria, actinomycetes and fungi in soils with transgenic papaya increased 1.6-4.46, 0.63-2.5 and 0.75-2.30 times. RP-transgenic papaya and non-transgenic papaya produced significantly different enzyme activities in arylsulfatase (5.4 5.9x), polyphenol oxidase (0.7-1.4x), invertase (0.5-0.79x), cellulase (0.23 0.35x) and phosphodiesterase (0.16-0.2x). The former three soil enzymes appeared to be more sensitive to the transgenic papaya than the others, and could be useful parameters in assessing the effects of transgenic papaya. Transgenic papaya could alter soil chemical properties, enzyme activities and microbial communities. PMID- 17078554 TI - Toxic effects of acetochlor and methamidophos on earthworm Eisenia fetida in phaiozem, northeast China. AB - Acetochlor and methamidophos are two important agrochemicals which are widely applied to agricultural production in northeast China. The investigation on the earthworm Eisenia fetida as an important type of soil animals exposed to single and binary-combined contamination of acetochlor and methamidophos was thus carried out. The single toxic effect test showed that the two agrochemicals had their toxicity to the earthworms living in phaiozem. Acetochlor had a stronger acute toxic effect on the earthworms than methamidophos. The mortality of the earthworms exposed to individual acetochlor and methamidophos changed with an increase in the exposure time and the exposed concentrations. The LD50 value of acetochlor and methamidophos toxic to the earthworms was 115.6-275.3 and 29.5 228.6 mg/kg, respectively. The weight of the earthworms was a more sensitive index compared to the mortality in indicating toxic effects of acetochlor and methamidophos in phaiozem. When considering both the mortality and the body weight change, the combined pollution of acetochlor and methamidophos in phaiozem resulted in their synergic toxic effects on the earthworms. PMID- 17078555 TI - An investigation of cellular distribution of manganese in hyperaccumlator plant Phytolacca acinosa Roxb. using SRXRF analysis. AB - Phytolacca acinosa Roxb. (P. acinosa) is a recently discovered manganese hyperaccumulator plant from southern China. It is a good candidate for phytoremediation of manganese(Mn) polluted soil for its high biomass and fast growth. Knowledge of the tissue localization and identification of heavy metals can provide essential information on metal toxicity and bioaccumulation mechanisms. Synchrotron radiation X-ray fluorescence spectroscopy (SRXRF) microprobe was used in this study to investigate the cellular distributions of Mn and other elements in root, stem, leaf, petiole and midrib of P. acinosa. The highest Mn content was found in the vascular tissues of root, stem, petiole and midrib. Cortex in root played a key role in Mn absorption and Mn was limited in the vascular bundle during the process of transportation in stem. Moreover, Mn content in leaf epidermis was higher than that in mesophyll, which suggested that the sequestration of Mn in leaf epidermis might be one of the detoxification mechanisms of P. acinosa. The significance of other elemental (such as P, S, K, Ca, Fe, Zn and Cu) distribution patterns and the correlation with Mn were also discussed. PMID- 17078556 TI - Dynamics of soil inorganic nitrogen and their responses to nitrogen additions in three subtropical forests, south China. AB - Three forests with different historical land-use, forest age, and species assemblages in subtropical China were selected to evaluate current soil N status and investigate the responses of soil inorganic N dynamics to monthly ammonium nitrate additions. Results showed that the mature monsoon evergreen broadleaved forest that has been protected for more than 400 years exhibited an advanced soil N status than the pine (Pinus massoniana) and pine-broadleaf mixed forests, both originated from the 1930's clear-cut and pine plantation. Mature forests had greater extractable inorganic N pool, lower N retention capacity, higher inorganic N leaching, and higher soil C/N ratios. Mineral soil extractable NH4(+) N and NO3(-)-N concentrations were significantly increased by experimental N additions on several sampling dates, but repeated ANOVA showed that the effect was not significant over the whole year except NH4(+)-N in the mature forest. In contrast, inorganic N (both NH4(+)-N and NO3(-)-N) in soil 20-cm below the surface was significantly elevated by the N additions. From 42% to 74% of N added was retained by the upper 20 cm soils in the pine and mixed forests, while 0%-70% was retained in the mature forest. Our results suggest that land-use history, forest age and species composition were likely to be some of the important factors that determine differing forest N retention responses to elevated N deposition in the study region. PMID- 17078557 TI - Dissipation of chlorpyrifos in pakchoi-vegetated soil in a greenhouse. AB - The dissipation of chlorpyrifos in pakchoi-vegetated soil was investigated in the summer and autumn in a greenhouse and field, respectively. The dissipation of chlorpyrifos in pakchoi-grown soil was comparatively described by fitting the residue data to seven models (1st-order, 1.5th-order, 2nd-order, RF 1st-order, RF 1.5th-order, RF 2nd-order, and bi-exponential or two-compartment models). Statistical analysis was performed using the SPSS 11.5 statistical package. The bi-exponential model was selected as the optimal model according to the coefficient of determination r2. The dissipation half-lives (DT50) of chlorpyrifos in pakchoi-vegetated soil at the recommended dose in the summer and autumn, calculated by the bi-exponential model, were 0.6 and 1.2 d in a greenhouse, 0.4 and 1.0 d in a field, respectively; the corresponding values at double dose were 1.2 and 2.1 d in a greenhouse, 0.5 and 1.3 d in a field, respectively. The kinetic data indicate the dissipation of chlorpyrifos in pakchoi-grown soil in a greenhouse is slower than that in a field, and dissipates slower in the autumn than in the summer. PMID- 17078558 TI - Microwave-assisted regeneration of hypercrosslinked polymeric adsorbent saturated with nitrophenols. AB - Thermal regeneration of hypercrosslinked polymeric adsorbent (HPA) was performed at 67 degrees C. Experiments were carried out using a multimode microwave oven operating at 2450 MHz and a thermostatic water bath so as to compare the effect of the different heating mechanisms on the regeneration efficiency of the HPA saturated with nitrophenols. The temperature rise action of the heterogeneous regeneration system under microwave irradiation was described. Additionally, a complete structure and chemical characterization of the regenerated HPA was also carried out in order to study the influence of the subsequent regeneration cycles on the structure and the adsorption capacities of the adsorbents. Structure characterization of the regenerated HPA was accomplished by means of the analysis of FTIR spectra. PMID- 17078559 TI - Photodegradation of bisphenol A in Fe(III)-oxalate complexes solution. AB - The aqueous photodegradation of bisphenol A (BPA) in the presence of Fe(III) oxalate complexes (Fe(III)-Ox), which are common compositions of natural water, was investigated in this study. BPA underwent rapid indirect photolysis in Fe(III)-Ox solution under simulated solar irradiation, proceeding pseudo-first order kinetics. The photolysis rate increased with decreasing pH or initial BPA level and increasing Fe(III)/oxalate concentration ratio. Hydroxyl radicals (*OH), which were generated from the photochemical processes of Fe(III)-Ox complexes and contributed to the photooxidation of BPA, were determined by molecular probe and electron spin resonance (ESR) methods with the steady-state concentration of 2.56 x 10(-14) mol/L. Superoxide anion radical (O2*-) was considered as the precursor of *OH and qualitatively determined by adding nitro blue tetrazolium as well as ESR experiments. Based on the structural analysis of the intermediate photoproducts of BPA in Fe(III)-Ox complexes solution, the possible degradation pathways of BPA were proposed, involving *OH addition, alkyl scission and alky oxidation. The results indicate that the photochemical reactivity of Fe(III) may affect the environmental fate of BPA in natural water significantly. PMID- 17078560 TI - Dependence of bisphenol A photodegradation on the initial concentration of oxalate in the lepidocrocite-oxalate complex system. AB - To understand the degradation of endocrine disrupting chemicals (EDCs) in natural environment with existence of iron oxides and carboxylic acids, the dependence of bisphenol A (BPA) photodegradation on the initial concentration of oxalate (COX) in lepidocrocite (gamma-FeOOH) aqueous suspension was investigated under both UV and visible lights in this study. Lepidocrocite powder was home-prepared by a hydrothermal process. It was found that BPA degradation was promoted greatly in the presence of oxalate owing to the formation of lepidocrocite-oxalate complex. And there was an optimal COX, which was 2.0 and 2.4 mmol/L, under UV and visible lights, respectively. The first-order kinetic constant, k value increased 38 times from 0.17 x 10(-2) min(-1) in the absence of oxalate to 6.39 x 10(-2) min( 1) in the presence of oxalate with an optimal COX (2.0 mmol/L) under UV irradiation, and almost 306 times from 0.02 x 10(-2) min(-1) in the absence of oxalate to 6.11 x 10(-2) min(-1) in the presence of oxalate with an optimal COX (2.4 mmol/L) under visible irradiation. The BPA degradation rate increased and the first-order kinetic constants decreased with the increase in BPA initial concentration. The dependence of the variation of pH value, total-Fe and Fe2+ during the photoreaction on COX was also investigated. The pH value increased obviously with the reaction time. Total-Fe increased dramatically at the first 5 min and then decreased quickly under UV irradiation and slowly under visible irradiation. The initial concentration of oxalate is a main factor to affect BPA photodegradation in aqueous suspension under both UV and visible lights. PMID- 17078561 TI - Photo-degradation of acid-red 3B dye catalyzed by TiO2 nanotubes. AB - TiO2 nanotube precursor was synthesized by the hydrothermal reaction of TiO2 powders with NaOH solution and the properties of the nanotube materials were tuned using different post-treatments. Transmission electron microscopic (TEM) observation revealed that the nanotube could be obtained by either a direct rinse with acid solution or rinse with distilled water followed by acid solution. The results of X-ray diffraction (XRD) and inductively coupled plasma (ICP) analysis indicated that the nanotube material was composed of H2Ti2O5 x H2O. In addition, the photocatalytic activities of the resulting catalysts were found to be strongly dependent on the post-treatment. The results of the photocatalytic reaction showed that the degradation of Acid-red 3B dye fitted pseudo-zero-order kinetics and TiO2 nanotube prepared under direct rinse with acid solution exhibited a higher catalytic efficiency compared to other catalysts. PMID- 17078562 TI - Expression of c-fos and oxidative stress on brain of rats reared on food from mercury-selenium coexisting mining area. AB - Wanshan mercury mine is the largest mercury deposit in Guizhou Province of China, but there were few reports on mercury toxic effect in the mining area. In order to study the neurotoxicity of food from Wanshan mercury mine area and probe into the effect of food from Wanshan mercury miner area on the changes of brain oxidative damage and expression of c-fos gene. The rats were exposed to mercury contaminated food for 20 d. The content of malondialdehyde (MDA), superoxide dismutase (SOD), GSH-peroxidase (GSH-px) and Glutathione (GSH) in rat brain was measured, and the effect of mercury contaminated rice on the expression of c-fos mRNA in rat brain and the expression of c-FOS protein in cortex, hippocampus were observed using reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemical methods. The results showed the levels of GSH, MDA, SOD and of GSH-dependent enzymes in the rat brain changed between exposure groups and control group; The mercury polluted rice induced significantly the expression of c-fos mRNA; the c-FOS positive cells in hippocampus and cortex of exposure groups were significant different from control group (P<0.01). It could be concluded that oxidative stress signals could contribute to the induction of immediate early genes (IEGs); free radicals and their by-products might not only cause oxidative damage, but also influenced gene expression; IEGs c-fos participated in the toxicity process of brain injury by mercury polluted food. PMID- 17078563 TI - Biodegradability of four phthalic acid esters under anaerobic condition assessed using natural sediment. AB - Biodegradability of di-n-butyl phthalate (DBP), butylbenzyl phthalate (BBP), di ethylhexyl phthalate (DEHP), and di-isononyl phthalate (DINP) under an anaerobic condition was evaluated using three natural sediment microcosms obtained from ponds in Osaka, which had not been significantly polluted by the chemicals. The degradabilities of the four phthalic acid esters (PAEs) were analyzed by a first order kinetic model with a lag phase and ranked as DBP>BBP>>DEHP>DINP. The PAEs with shorter alkyl-chains, DBP and BBP, were degraded with quite short lag phases near to zero and short half-lives of a few days. The PAEs with longer alkyl chains, DEHP and DINP, were degraded with lag phases of 5-30 d and the quite long half-lives of a couple of hundred days. Although no data was available on the anaerobic biodegradability of DINP before this study, it was clarified that DINP can be degraded with slow degradation rates. The fact that all the three intact sediments were capable of biodegradation of the PAEs suggests that potential of anaerobic biodegradation of PAEs is widespread in the aquatic environment. PMID- 17078564 TI - Degradation of malathion by Pseudomonas during activated sludge treatment system using principal component analysis (PCA). AB - Popular descriptive multivariate statistical method currently employed is the principal component analyses (PCA) method. PCA is used to develop linear combinations that successively maximize the total variance of a sample where there is no known group structure. This study aimed at demonstrating the performance evaluation of pilot activated sludge treatment system by inoculating a strain of Pseudomonas capable of degrading malathion which was isolated by enrichment technique. An intensive analytical program was followed for evaluating the efficiency of biosimulator by maintaining the dissolved oxygen (DO) concentration at 4.0 mg/L. Analyses by high performance liquid chromatographic technique revealed that 90% of malathion removal was achieved within 29 h of treatment whereas COD got reduced considerably during the treatment process and mean removal efficiency was found to be 78%. The mean pH values increased gradually during the treatment process ranging from 7.36-8.54. Similarly the mean ammonia-nitrogen (NH3-N) values were found to be fluctuating between 19.425 28.488 mg/L, mean nitrite-nitrogen (NO3-N) ranging between 1.301-2.940 mg/L and mean nitrate-nitrogen (NO3-N) ranging between 0.0071-0.0711 mg/L. The study revealed that inoculation of bacterial culture under laboratory conditions could be used in bioremediation of environmental pollution caused by xenobiotics. The PCA analyses showed that pH, COD, organic load and total malathion concentration were highly correlated and emerged as the variables controlling the first component, whereas dissolved oxygen, NO3-N and NH3-N governed the second component. The third component repeated the trend exhibited by the first two components. PMID- 17078565 TI - Partial least square modeling of hydrolysis: analyzing the impacts of pH and acetate. AB - pH and volatile fatty acids both might affect the further hydrolysis of particulate solid waste, which is the limiting-step of anaerobic digestion. To clarify the individual effects of pH and volatile fatty acids, batch experiments were conducted at fixed pH value (pH 5-9) with or without acetate (20 g/L). The hydrolysis efficiencies of carbohydrate and protein were evaluated by carbon and nitrogen content of solids, amylase activity and proteinase activity. The trend of carbohydrate hydrolysis with pH was not affected by the addition of acetate, following the sequence of pH 7>pH 8>pH 9>pH 6>pH 5; but the inhibition of acetate (20 g/L) was obvious by 10%-60 %. The evolution of residual nitrogen showed that the effect of pH on protein hydrolysis was minor, while the acetate was seriously inhibitory especially at alkali condition by 45%-100 %. The relationship between the factors (pH and acetate) and the response variables was evaluated by partial least square modeling (PLS). The PLS analysis demonstrated that the hydrolysis of carbohydrate was both affected by pH and acetate, with pH the more important factor. Therefore, the inhibition by acetate on carbohydrate hydrolysis was mainly due to the corresponding decline of pH, but the presence of acetate species, while the acetate species was the absolutely important factor for the hydrolysis of protein. PMID- 17078566 TI - Influence of temperature on performance of anaerobic digestion of municipal solid waste. AB - The influence of temperature on the performance of anaerobic reactors for treating the organic fraction of municipal solid waste (OFMSW) was studied. Batch digestion of OFMSW was carried out for 32 d at different temperature (25 degrees C, 35 degrees C, 45 degrees C and 55 degrees C) conditions for total solid concentrations (TS) 17% with the ratio of total organic carbon to nitrogen (C/N) being, 25:1 respectively, while keeping other parameters constant such as inoculum, start-up pH, reactor volume (2 L) and so on. Temperature can influence the methanogenic bacteria activity, accordingly inhibiting the OFMSW biodegradation and stabilization efficiency. Anaerobic reactors excelled at TS reduction, total volatile solid reduction, chemical oxygen demand reduction, increasing cumulative biogas production, whose rate was at temperature (35 degrees C and 55 degrees C) conditions. Methane concentration in the biogas was above 65% in four reactors. In addition, the fluctuation of temperatures resulted in the biogas production variation. The data obtained indicated that temperature had a significant influence on anaerobic process. PMID- 17078567 TI - Phosphorus accumulation and eutrophication in feed-supply freshwater fishponds. AB - The rapid growth and intensification of freshwater fishery can cause imbalances between phosphorus (P) input in feed and its output in produce. This aquaculture can result in enriching exogenous P in fishponds and, consequently, accelerates the process of eutrophication. To assess relations among input, accumulation, release of P and as a consequence degrading water quality in terms of chlorophyll a (Chl-a) in freshwater fishponds, fourteen fishponds with feed supply, nine fishponds without feed supply, and five non-fish ponds in Shaoxing Plain, southeast China were selected for comparing P accumulation in their waters and surface sediments. Surface sediment samples were collected from each pond to evaluate their total P, water soluble P, Olsen P, algal available P, and P fractions. Water samples were also collected from the ponds to measure concentrations of dissolved P and Chl-a. Total P in the sediments ranged from 0.88 to 1.73 g/kg in the fishponds with feed supply, that in the non-fish ponds ranged from 0.47 to 0.86 g/kg. Organic P, accounted for 23% to 60% of total P in the sediments, was an important P fraction and increased linearly with increasing organic matter. Long-term application of feeds resulted in increased P availability in the bottom sediments and degradation of water quality in the freshwater fishponds. Compared with non-fish ponds, sediments from the feed supplied fishponds contained considerably higher Olsen P, algal available P, and water soluble P. Higher proportions of the labile P (NH4Cl-P) and potentially labile P (NaOH-IP) were also found in the sediments from the fishponds. High solubility of P in the sediments resulted in elevation of P and chlorophyll-a concentration in the pond water. The dissolved P concentration in the pond water increased in the order of non-fish ponds (12 microg/L) < fishponds without feed supply (24 microg/L) < fishponds with feed supply (66 microg/L). Linear correlations between concentrations of total P, Olsen-P, algal available P, water soluble P and P concentration in saturation extracts in the sediments and dissolved P in the pond water indicated that there was a buffering action of the sediment constituents on the dissolved P. PMID- 17078568 TI - Gridded inventories of historical usage for selected organochlorine pesticides in Heilongjiang River Basin, China. AB - The use of technical HCH (1, 2, 3, 4, 5, 6-hexachlorocyclohexane), DDT (dichlorodiphenyltrichloroethane) and lindane in Heilongjiang River Basin (HRB) of China was studied. Between 1952 and 1984 the total usage in the HRB was 108900 t for technical HCH, and 4900 t for DDT, respectively, and the use of DDT due to dicofol application in the HRB was 220 t from 1984 to 2003. The usage of lindane in HRB was 109 t from 1991 to 2000. The results show that the highest technical HCH use (41800 t) in HRB was on maize, accounting for 38.3% of the total usage on all crops, followed by the use on wheat (28000 t, 25.7%) and on soybean (26600 t, 24.4%). The highest DDT use (2300 t) was on soybean, accounting for 46.8% of the total usage on all crops, followed by the use on maize (1500 t, 31.3%) and on sorghum (600 t, 13.2%). The major use of lindane was mainly on wheat. Gridded usage inventories of these three OCPs (organochlorine pesticides), in HRB with a 1/6 degrees latitude by 1/4 degrees longitude resolution have been created by using different gridded cropland as surrogates. Some soil samples have also been collected in HRB. Data of usage and soil concentrations of sigma HCH in HRB match quite well to those in Shanghai region, and much higher soil concentration of sigma DDT in Shanghai region could be due to much heavier dicofol use in this region. PMID- 17078569 TI - Prediction of the concentration of chlorophyll-a for Liuhai urban lakes in Beijing City. AB - The weekly water quality monitor data of Liuhai lakes between April 2003 and November 2004 in Beijing City were used as an example to build an artificial neural networks (ANN) model and a multi-varieties regression model respectively for predicting the fresh water algae bloom. The different predicted abilities of the two methods in Liuhai lakes were compared. A principle analysis method was first used to select the input variables of the models to avoid the phenomenon of collinearity in the data. The results showed that the input variables for the artificial neural networks were T, TP, transparency(SD), DO, chlorophyll-a (Chl a), pH and the output variable was Chl-a. A three layer Levenberg-Marguardt feed forward learning algorithm in ANN was used to model the eutrophication process of Liuhai lakes. 20 nodes in hidden layer and 1 node of output for the ANN model had been optimized by trial and error method. A sensitivity analysis of the input variables was performed to evaluate their relative significance in determining the predicted values. The correlation coefficient between predicted value and observed value in all data and in test data were 0.717 and 0.816 respectively in the artificial neural networks. The stepwise regression method was used to simulate the linear relation between Chl-a and temperature, of which the correlation coefficient was 0.213. By comparing the results of the two models, it was found that neural network models were able to simulate non-linear behavior in the water eutrophication process of Liuhai lakes reasonably and could successfully estimate some extreme values from calibration and test data sets. PMID- 17078570 TI - [Chronological evolution of the renal transplant in Spain]. AB - INTRODUCTION: We try to establish the chronological evolution that has taken the renal transplant in Spain. Supported on the heuristic one we gather from the first experiences and scientific works published at the beginning of the sixties up to those of the finals of the XXth century in which the procedure is already consolidated. MATERIAL AND METHOD: We check the Spanish bibliography urology, the publications of renal transplant, those of dialysis and the nefrology and all the bibliographical references we have located beside contributing our personal experience. RESULTS: The zeal to help the patients in renal terminal insufficiency was the engine that stimulated the Spanish urologist to begin with the practice of the renal transplants. In May, 1960 the Dr. Martinez-Pineiro carries out the first one, without satisfactory result and is in July, 1965 when the team of the Dr. Gil-Vernet realizes a transplant of corpse with long billiard cue sobrevida of the patient and of the organ. With the advances in the treatments immunosuppressants, the procedure is consolidated as the best option for the renal insufficiency treatment and the Spain principal hospitals add to the transplants. CONCLUSIONS: The Spanish urologist, with their effort, dedication and confidence in the results of the technology, are the beginning and principal persons in charge of the surgical activity of the implant and only when the benefit was evident the Administration moved to establishing the procedure for its development; after the constitution of the ONT, with its unfolding of means, they achieves that Spain reaches and guarantees a world primacy so much in number of donors as of implants. PMID- 17078571 TI - [Incidental prostatic cancer: T1a-T1b. Our experience after observation/radical surgery and literature review]. AB - OBJECTIVES: To analyse the progress of T1a and T1b prostate cancer diagnosed in our hospital. MATERIAL AND METHODS: Retrospective study of 40 patients in T1a-T1b clinical stage diagnosed with prostate adenocarcinoma in our hospital, from 1986 to 1999. A restaging biopsy was performed on the 16 T1a patients after initial diagnosis and control. A radical prostatectomy was performed on the 24 T1b patients. They were all monitored every six months with rectal exam and PSA. We analysed biological and/or clinical progression, time to progression, mortality caused by the tumour and survival. RESULTS: None of the 16 patients with T1a clinical stage presented tumour progression, with a median follow-up of 90 months. 12,5% of the 24 T1b cases presented tumour progression, with a median follow-up of 70 months. Cancer-specific mortality was one patient (4,16 %) in the T1b group. CONCLUSIONS: Observation and follow-up with PSA and rectal exam appears to be a good option for T1a clinical stage, given the good prognosis. Our results show that patients with T1a clinical stage and good prognostic factors could be at a similar risk of suffering from a new prostate cancer as the normal population, although prospective studies are required to validate these results. T1b cases require active treatment and closer monitoring. PMID- 17078572 TI - [Immunohistochemical expression of p53, p21, p16, and cyclin D1 in superficial bladder cancer. A tissue microarray study]. AB - INTRODUCTION AND OBJECTIVES: To retrospectively assess the relationship between immunohistochemical expression of p53, p21, p16, and cyclin D1, with recurrence, progression and survival in superficial bladder cancer. METHODS: 163 patients undergoing transurethral resection for superficial bladder cancer between February 1995 and March 2004. Tumor samples were included in a tissue microarray support that was serially sectioned for immunohistochemical staining. Grade and stage associations for each marker were evaluated by the Chi-square test. Assessment of the relationship with recurrence, progression, and survival Kaplan Meier curves and log-rank test were used. RESULTS: There were no statistically significant differences in marker expression depending on tumor grade and stage, with the exception of Cyclin D1, that was significantly different depending on tumor stage (p=0.030). p21 expression was related to tumor recurrence (p=0.035), progression (p=0.008) and survival (p=0.034). p16 expression was also related to recurrence (p=0.048) and survival (p=0.047), but not to tumor progression (p=0.116). p53 and Cyclin D1 were not statistically associated with tumor recurrence, progression or survival. CONCLUSIONS: In our experience, only p16 and p21 may be useful in the management of superficial bladder tumors, as they are predictors of recurrence and survival in Ta and T1 patients. PMID- 17078573 TI - [Stage pT0 bladder tumors after radical cystectomy: a review of our series]. AB - OBJECTIVES: To evaluate the data of progression and survival in 43 patients who underwent cystectomy with stage pT0 according to classification TNM-2002. MATERIALS AND METHODS: between 1988 and 2003 in our center had realized 420 cystectomies, 43 patients (10.2%) had not tumor in the cystectomy specimen. RESULTS: In these 43 cases the initial clinical stage (in the transuretral resection of bladder) was T1 in 10 cases (23,3 %), T2 in 31 cases (72%) and T3 in 2 cases. As far as the degree 24 patients presented G2 (55.8%) and 19 (44.2%) were G3. Median time from the transuretral resection to the cistectomy was of 44 days at a median follow-up of 89.3 months. Progression-free survival in the 43 patients was of 180,6 months, but during the follow-up it appeared progression in 7 patients, with disease free survival at 36 months (3-126), since the date of the cistectomia. During the follow up, 5 patients died. When we analyzed the cancer-specific survival according to tumor stage, for the T2 with an average cancer-specific survival is of 180 months, decreasing to 35 months considerably for T3. Similar it happens with the degree of differentiation, significantly diminishing as it advances the degree, with an average of cancer-specific survival for the G3 at 122.6 months. In the same way it happens with pathological positive lymph nodes in the radical cistectomy, with a cancer-specific survival of 188 months when it is N0 and of 54 months if the adenopathy was positive (N+). CONCLUSION: In our experiencie urothelial carcinoma pT0 present a prolonged free period of disease (medium of 180 months). The associated factors of risk to a smaller free period of disease are high degree of differentiation (G3, 116 months), the infiltration of deep layers in the transuretral resection (T3, 32 months) and the ganglionary affectation (pN+ 45 months). PMID- 17078574 TI - [Solid renal mass up to 4 cm. Analysis of the diagnostic procedures, TNM staging and surgical treatment]. AB - OBJECTIVES: We perform a retrospective review of renal tumors operated with a maximum size of 4 cm to determine if the diagnostic methodology was adequate, the TMN staging prognostic accuracy (UICC 2002) and the goal surgical treatment. MATERIAL AND METHOD: Between 1984 to 2005, 78 renal units form 74 patients (4 bilateral synchronous) operated at the Service of Urology of the Hospital Aleman de Buenos Aires. Age distribution, sex, presentation form (incidental and symptomatic), diagnostic methodology, laboratory variables, surgical treatment (partial or radical surgery), histopathology, Fuhrman grade, tumor size, bilaterality, multicentricity, TNM staging, evolution and survival (Kaplan Meier) were analyzed. RESULTS: 78 renal units from a total of 192, 40.62% were analyzed. The median age was 58.72 years. 69% were male and 31% female. The presentation form was 91.90 % incidental and symptomatic 8.10% (Hematuria in 5 and lung metastasis in 1). The diagnosis was performing with ultrasound and CT scan 63 cases, in 4 was also used resonance magnetic imaging (RMI) and in 7 ultrasound and RMI. Arteriography was used in 11 cases, 3 showed tumor and 8 were normal. Biopsy was performing in 5 cases, all positive for clear cells carcinoma. Globular sedimentation was the only one laboratory abnormality in 12 cases. Surgical treatment was radical surgery in 35 renal units (44.87%) and conservative surgery in 43 units (55.13%). Pathology clear cells carcinoma (CCC) 79.48%, papillary carcinoma 1.28%, angiomyolipoma (AML) 8.97%, oncocytoma 7.69% and adenoma 2.56%. The Fuhrman grade was 1 in 76.19%; 2 in 20.63% and 3 in 3.18%. Bilateral tumor were found in 4 cases 2 CCC, 1 CCC and AML and 1 CCC and adenoma. Tumor median size was 2.93 cm. Staging was T1a 96.82%, T3a 1.59% and T3aM+ 1.59%. Follow-up could be made in 54 of 61 cases. At median follow-up of 52.25 months, 50 cases were disease free, 3 died by progression at 18, 33 and 82 months and all of them were symptomatic tumors, 1 died by a non related cause. Survival rate was 94%. CONCLUSIONS: 1)Ultrasound and CT scan obtained a highest diagnostic accuracy for solid renal mass. Biopsy in selective cases could contribute to achieve a correct treatment strategy. 2) Conservative surgery was the goal treatment in selected tumors up to 4 cm. and we believe that TNM staging should contemplate the presentation form to improve the prognostic value. PMID- 17078575 TI - [Modificated plication technique of the tunica albuginea in penis curvature correction]. AB - We present our penile curvature treatment experience in Peyronie's disease and cogenital curvature, using a modificated plication technique of the tunica albuginea with Prolene inverted sutures covered with Vicryl ones. The first suture gives security to the plication and the second one hides the first dots avoiding the nonabsorbable sutures bother. PMID- 17078576 TI - [Efficacy and safety of two dosing regimens with Tadalafil in Spanish men with erectile dysfunction: results from the SURE study in 14 European countries]. AB - OBJECTIVE: [corrected] To compare the efficacy and safety of tadalafil 20 mg administered 3 times/week (SCH) vs. on demand (OD) in a cohort of Spanish men with erectile dysfunction (ED), since Tadalafil period of responsiveness lasts up to 36 hours post-dosing. MATERIAL AND METHODS: The 418 Spanish patients participating in the European multicenter, crossover, open-label SURE clinical trial (comprising 4262 men) were randomly assigned to one of the treatment sequences: tadalafil 20 mg SCH for 5-6 weeks followed by tadalafil 20 mg OD for 5 6 weeks, or the inverse sequence. At completion, patients were asked to select the regimen they preferred to receive in an extension phase. RESULTS: In both regimens, tadalafil led to a similar improvement in erectile function compared to baseline. However, the SCH regimen showed statistically significant higher scores for several IIEF questions (i.e. sexual desire domain). Normal erectile function (IIEF EF domain score > or = 26) was achieved by 69.3% of patients on SCH and 64.3 % on OD, with a sexual intercourse success rate (SEP3) of 75.6% and 72.2% respectively (p<0.05). Nevertheless, more patients preferred to receive tadalafil OD for the extension phase (55.9% vs 44.1%, p<0.05). Tadalafil was well tolerated in both regimens. The most common TEAEs (> or = 5%) were headache, dyspepsia and back pain. There were no clinically significant differences in the incidence of TEAEs between regimens. CONCLUSIONS: Tadalafil 20 mg is efficacious and well tolerated for the treatment of ED, regardless the regimen of administration (OD or SCH). Patients can choose the pattern of administration that fits better with their expectations. PMID- 17078577 TI - [Regenerative medicine in andrology: tissue engineering and gene therapy as potential treatment options for penile deformations and erectile dysfunction]. AB - Tissue engineering and gene therapy are currently investigated in animal studies for reconstructing penile tissue or treating erectile dysfunction. This review aims to ecamine these experimental efforts from the last years and tries to give a brief introduction to the basic methodology of these new techniques from the field of regenerative medicine. PMID- 17078578 TI - [Complications in laparoscopic nephrectomy]. AB - OBJECTIVE: To report the complications and their respective management in our series of laparoscopic nephrectomy. MATERIAL AND METHODS: Between November 1992 and March 2004, 319 patients underwent laparoscopic ablative renal surgery: 152 were women and 167 were men, mean age was 45 years (range: 0,5-82 years). There were 70 radical nephrectomies, 16 radical nephroureterectomies, 63 partial nephrectomies, 124 simples nephrectomies, 15 simples nephroureterectomies, 8 heminephrectomies and 23 living donor nephrectomies. RESULTS: The complication rate was 5.64%. The most frequent complications were bleeding (2.5%) and retroperitoneal haematoma (1.2%). There was not statically difference in complications rate by age, sex, or approach via: transperitoneal versus retroperitoneal. Complication rate was alike with laparoscopic or hand assisted technique. There was not mortality in the series. CONCLUSIONS: We have a low complication rate in laparoscopic nephrectomy. We believe that laparoscopy is a safe technique for patients that require ablative renal surgery when the surgical team has experience in advanced laparoscopic surgery. PMID- 17078579 TI - [Use of a virtual immersion computer simulator as a model for basic training in laparoscopic urology]. AB - BACKGROUND: to date, it has not been defined the best method for teaching urologic laparoscopy, however it is well recognized that it involves a steep learning curve. METHODS: A course of Laparoscopic Urology was done in our Institute. The program included skill practices in a virtual immersion simulator which evaluated, the score and time to complete each activity. This was done in a group of residents with previous experience with this virtual simulator (group 1) and another group of residents with no experience (group 2). Four different basic tasks were performed in the virtual simulator, which included: coordination, cutting, clip application and performing a simple suture. RESULTS: When we compared the scores between both groups the mean scores for each task were superior in group 1 compared to the group 2, with no statistically significant difference, however when we compared the time to complete each task, it was shorter in group 1 compared to group 2 with a statistically significant difference. CONCLUSIONS: The performance of residents without experience in a virtual simulator was similar to that of previously trained residents, however it takes less time to complete each task as the resident gains experience in these simulators. The use of virtual simulators for laparoscopy training are useful when learning basic techniques allowing the surgeon to improve hand dexterity and coordination in laparoscopic surgery. PMID- 17078580 TI - [Multiple schwannoma of the penis]. AB - Schwannoma or neurilemmoma designate a tumour that originates in the Schwann cells of peripheral nerves, therefore occurring anywhere in the body. They are very rare in the penis. In the literature are reported twenty seven cases of penile schwannoma. We report a case of multiple schwannoma of the penis and make a brief literature revision about this rare pathology. PMID- 17078581 TI - [Papillary carcinoma after 13 years of correct function of renal graft]. AB - It is known the greater probability appearance of malignancy injuries in patients with renal graft due to its inmunosupresion. We expose a case in which after thirteen years of correct operation of the renal transplant a tumor is diagnosed of accidental form by means of ultrasonography of graft's control. It was demonstrated by percutaneous biopsy that it was a carcinoma to papilar and later transplanctectomy was made. We raised a reflection about the novo tumors on renal graft given to the high number of patients with funcionante transplant during long years and the little evidence in Literature, proposing a possible registry of such valuing its behavior and comparing it with the well-known ones on native kidneys without inmunosupresion situation. PMID- 17078582 TI - [Prostate adenocarcinoma metastatic to penis]. AB - Prostate cancer is a disease that appears with a very high frequency. However, the presence of metastasis to the penis is rare. Only 98 cases reported in the literature up to 2003. We present one case of a patient with painless metastatic nodules on the penis secondary to a prostate cancer. PMID- 17078583 TI - [Small cell neuroendocrine tumour of the bladder: with reference to a case and bibliographical revision]. AB - INTRODUCTION AND OBJECTIVES: The small cell neuroendocrine tumour is an infrecuent neoplasia, with inmunohistochemistry being the key to diagnosis. We present a new case making reference to treatment and its evolution there after. MATERIAL AND METHODS: The clinic, diagnosis and treatment of this tumour is described. Bibliographical revision follours. CONCLUSIONS: The neuroendocrine tumour of small cell is an infrecuent neoplasia, in which the inmunohistochemistry study is key in the diagnosis. The differential diagnosis includes the high degree diferentiation transitionals cells carcinoma and primary and secondary linfoma. The standard treatment is based on chemotherapy plus surgery. PMID- 17078584 TI - [Granulomatous pneumonitis secondary to BCG bladder instillation]. AB - We report two cases of pulmonary BCG-induced pneumonitis recently diagnosed amongst our patients. The first case is a 81 year old man under BCG bladder instillation treatment because of high grade superficial bladder cancer who developed a severe interstitial granulomatous pneumonitis. The patient was treated with corticosteroids and tuberculostatics with a rapid and complete response. The second patient is a young man who presented only with persistent fever and also had a complete response after treatment. PMID- 17078585 TI - [In reference to the article "Epidemiological situation of prostate cancer in Spain]. PMID- 17078586 TI - [Lithiasis in ureterocele]. PMID- 17078587 TI - [Severe condylomatosis treated with CO2 laser]. PMID- 17078588 TI - [Silicone autoinjection in penis]. PMID- 17078589 TI - [Symptomatic vesical lithiasis]. PMID- 17078590 TI - [What is the frequency of cardiovascular risk factors and co-morbidity in patients with rheumatic diseases?]. AB - Analysis of national data from the health ministry programme of reduction of the cardiovascular risks (2002-2005) shows a high frequency of cardiovascular disease and cardiovascular risk factors in the general population. It is of interest to analyse these data in relation to the practice of rheumatology. In addition, the frequency of cardiovascular pathologies is higher in patients with rheumatoid arthritis and spondylarthropothy. These notions are also very important since these two populations are often treated with non-steroidal anti-inflammatory drugs for a long duration. General knowledge shown in this article concerning the cardiovascular risk factors and co-morbidities in the patients with rheumatic pathologies allows, within the context of a therapeutic decisional strategy in rheumatology, a better estimation of the individual benefit/risk ratio of each prescription and more particularly that of non-steroidal anti-inflammatory drugs. PMID- 17078591 TI - [What do we know about the cardiovascular toxicity of the NSAIDs?]. AB - Following the arterial thrombotic risk of rofecoxib (myocardial infarct and cerebral ischemic accidents) that led to its withdrawal from the market, the other coxibs then the NSAIDs have also been blacklisted. The factors responsible for the cardiovascular risk associated with the ingestion of the NSAIDs, selective or not, are not clearly identified. The objective of this review was to collect the available data from the literature, in order to better evaluate of the risk and its causes, principally on the basis of the results of randomised studies, but also of case reports and meta-analyses. There is an increase in the risk of arterial thrombotic events under coxibs and traditional NSAIDs, however the risk is variable for both classes. The cardiovascular risk linked to celecoxib seems variable and modest, and at a standard dose and for usual treatment durations, the risk is probably inexistant. While the real risk of classical NSAIDs is difficult to appreciate from the available results, it can be concluded that the cardiovascular risk of naproxen is low. While part of the cardiovascular consequences of rofecoxib could be associated with increased arterial pressure, these effects are not exclusive to the coxibs since they have been observed with the conventional NSAIDs. However the increase in arterial pressure cannot probably explain everything. Similarly the cardiac insufficiency associated more particularly with rofecoxib, especially in some groups of patients (very old subjects) is not a new type of complication and does not seem to be more frequent with coxibs than with classical NSAIDs. No short-term arterial thrombotic risk of the coxibs and NSAIDs has been clearly demonstrated. PMID- 17078592 TI - [Are there any differences in the cardiovascular tolerance between classical NSAIDs and coxibs?]. AB - Three placebo-controlled studies have demonstrated deleterious cardiovascular (CV) effects of rofecoxib, celecoxib, and pare/valdecoxib. It remains to be determined whether this CV toxicity is specific to coxibs, or shared with all non steroidal anti-inflammatory drugs (NSAIDs). Seven meta-analyses show that, in comparison with non-specific NSAIDs, the risk of thrombotic CV accident is increased with rofecoxib and celecoxib, but not with valdecoxib or lumiracoxib. Concerning the risk of thrombotic CV accident, seven of the ten observational studies which have evaluated the risk have found an increased risk for the non specific NSAIDs in comparison with non-exposed subjects. The seven observational studies, which evaluated the risk of coxibs, have all found an increased risk with rofecoxib, and two with celecoxib. Three studies out of six have shown on increase of risk with rofecoxib and one study out of five with celecoxib. Two of the three studies, which have compared rofecoxib with celecoxib, have found on increased risk with rofecoxib. Concerning the risk of arterial hypertension, aedemas or congestive cardiac insufficiency, a meta-analysis and a randomised trial have shown a deleterious effect of rofecoxib in comparison with celecoxib and non-specific NSAIDs. Two studies have shown a deleterious effect of the non selective NSAIDs and three a deleterious effect of rofecoxib in comparison with non-exposed subjects. Three studies have demonstrated a deleterious effect of rofecoxib in comparison with non-specific NSAIDs. No study has shown any deleterious effect of celecoxib in comparison with subjects non-exposed or exposed to non-specific NSAIDs. These studies suggest that all the NSAIDs, specific or not, increase the CV and renal risk. This risk seems variable from a compound to another one and must be evaluated, for each patient, according to the susceptibility and associated risk factors. While waiting for other long-term controlled studies, the available data show the existence of a risk of CV secondary effect linked to the class of NSAIDs, specific (coxibs) or not. PMID- 17078593 TI - [How to evaluate the cardiovascular and renal risk at the individual level?]. AB - The cardiovascular impact of the non-steroidal anti-inflammatory drugs and the higher cardiovascular mortality during treatment of inflammatory rheumatism impose a rigorous evaluation of the cardiovascular risk of rheumatic patients. Large epidemiological studies have identified risk factors for cardiovascular diseases such as the age, male gender, family history (infarct, stroke), tobacco consumption, systolic arterial pressure, renal insufficiency, hypercholesterolemia, diabetes mellitis, sedentariness, obesity and "electric" ventricular hypertrophy. Some equations make it possible to evaluate the absolute cardiovascular risk at the individual level, which corresponds to the onset risk of a stroke in the 10 years to come in a subject according to the number and importance of each of his risk factors. It has been demonstrated that the correction of one or more risk factors reduce the overall cardiovascular risk justifying the strategies for evaluating this risk to define therapeutic intervention thresholds. The impact of a long-term anti-inflammatory treatment or an inflammatory disease such as rheumatoid arthritis has not been the subject of specific epidemiological study allowing these elements to be included in an equation of the estimation of the cardiovascular risk. However, the introduction of on anti-inflammatory treatment, likely to increase the cardiovascular risk of a patient, certainly justifies an evaluation of the absolute cardiovascular risk. PMID- 17078594 TI - [How to watch over a patient treated with a NSAID in relation to the cardiovascular and renal risk?]. AB - Despite the cardiovascular risk attributable to the NSAIDs, these drugs are among the most prescribed treatments in the world. Recently to manage this risk during chronic inflammatory rheumatisms a surveillance plan has been developed based on a decision algorithm. Given that the arterial thrombotic risks (myocardial or cerebral) are observed essentially during long-term treatments, two types of situation have been envisaged, short-term treatments and long-term treatments. Before any short-term NSAID prescription (less than one month), the cardio-renal risk should be evaluated. A pre-therapeutic check-up should include the search for risk factors and the treatment surveillance should imply a clinical and biological check-up carried out after 2 to 3 weeks of treatment. Before any long term NSAID prescription (more than one month), the arterial thrombotic risk (cardiac or cerebral) should be evaluated and the patient's history determined. Pre-therapeutic check-up and surveillance should be carried out and reconsidered at least every three months. In the case of high risk the advice of a cardiologist or a nephrologist should be obtained and drastic therapeutic measures taken. There are cases where the prescription of NSAIDs or coxibs is possibly not recommended. PMID- 17078595 TI - [What are the cardiovascular complications of the analgesics and glucocorticoids?]. AB - When prescribing a non-steroidal anti-inflammatory treatment but also an analgesic or a glucocorticoid, the cardiovascular risk of the patient should be assessed. The analgesics have few cardiovascular side effects and the main complications observed are linked essentially to the vagal action of the opioids. Acetaminophen is considered by several scientific societies to be the first line analgesic treatment, particularly in case of cardiovascular risk but with caution since cardiovascular toxicity of acetaminophen cannot be totally excluded. An overdose of dextropropoxyphene can result in cardiotoxicity. On the other hand, the glucocorticoids need to be prescribed cautiously, at the lowest possible dose and for the shortest possible duration due to the non-negligible cardiovascular risk, hypertension, dyslipidemia, hypokaliemia. PMID- 17078596 TI - [Are the NSAIDs able to compromising the cardio-preventive efficacy of aspirin?]. AB - OBJECTIVES: Some studies have recently suggested a potential pharmacodynamic interaction between aspirin and some non-selective non-steroidal anti inflammatory drugs (NSAIDs). We have evaluated the reality of this pharmacodynamic interaction and analyse its clinical pertinence. METHODS: Literature review (Medline search - December 2005). RESULTS: Several ex vivo studies show that some non-selective NSAIDs can block the active site of Cox1 thus preventing aspirin from exerting its platelet anti-aggregating cardio preventive action. Cox2 selective molecules do not act at this site. The few studies, mainly case reports, have analysed the potential loss of the cardiovascular preventive benefit of aspirin in patients receiving concomitantly non-selective anti-inflammatory drugs with controversial results. IN PRACTICE: It seems necessary to know the existence of this pharmacodynamic interaction between aspirin at a low dose and some non-selective anti-inflammatory drugs notably ibuprofen and naproxen. In the absence of a clear clinical demonstration, it is advisable to avoid the non-selective NSAIDs in patients treated with a low dose of aspirin. It might be advisable to switch to an anti-aggregating treatment other than aspirin (clopidrogel, etc.) in these cases. At the present time, however, there are no data on which to base such a recommendation. PMID- 17078597 TI - [What is the impact of cardiovascular and renal complications on the benefit/risk ratio of the NSAIDs?]. AB - Long-term studies of NSAIDs have demonstrated cardiovascular toxicity which poses a general problem of the benefit risk between the relief of the patient's symptoms and the risk of digestive and cardiovascular toxicity. The digestive complications and the risk factors associated with the ingestion of NSAIDs have been defined by numerous studies but the prevention of this digestive toxicity can be attempted. The benefit-risk ratio caused by the digestive and cardiovascular toxicities of the NSAIDs and COX-2 inhibitors must be evaluated for each individual before starting a treatment. For a treatment of short duration in patients with a low risk of digestive and cardiovascular complications, the risk of digestive toxicity must be taken into consideration first and the COX-2 inhibitors are favoured. For a treatment of long duration the risk must be evaluated with the greatest care. In the case of cardiovascular risk factors, the cardiovascular risk must be evaluated and the treatment must be prescribed at the lowest dose for the shortest duration. In patients with very high risk of digestive and cardiovascular complications, since the digestive and cardiovascular extra-mortality associated with the ingestion of NSAIDs or COX-2 inhibitors is high, it is advised to abandon this type of long-term treatment. PMID- 17078598 TI - [Which information should be given to the patient concerning the cardiovascular and renal risk of the NSAIDs?]. AB - OBJECTIVE: Patient information is a topical subject. The aim of this review is to present the rare studies concerning the degree of patient information on the cardiovascular and renal risks of the non-steroidal anti-inflammatory drugs and to suggest some recommendations on this subject. METHODOLOGY: After analysis of the literature (Medline search - January 2006), a series of recommendations has been drown up following discussion among experts from different specialties (clinical epidemiology rheumatology cardiology nephrology gastroenterology). RECOMMENDATIONS: It appears necessary to warn all patients of the potential risk of hyperkolemia, renal insufficiency and/or hydrosodium retention (decompensation of an arterial hypertension or a cardiac insufficiency), especially subjects who present risk factors such as age over 75 years, a dehydration, a pre-existing renal disorder etc. Concerning the risk of arterial thrombotic incident (coronary or cerebral), it is important to indicate that this risk is rare and seems to be observed particularly during prolonged treatments and at high dose. PMID- 17078599 TI - Experimental and theoretical characterization of aluminum-based binary superatoms of AL12X and their cluster salts. AB - The geometric and electronic structures of aluminum binary clusters, AlnX (X = Si and P), have been investigated, using mass spectrometry, anion photoelectron spectroscopy, photoionization spectroscopy, and theoretical calculations. Both experimental and theoretical results show that Al12Si has a high ionization energy and low electron affinity and Al12P has a low ionization energy, both with the icosahedral structure having a central Si or P atom, revealing that Al12Si and Al12P exhibit rare-gas-like and alkali superatoms, respectively. Experiments confirmed the possibility that the change in the total number of valence electrons on substitution could produce ionically bound binary superatom complexes, the binary cluster salts Al12P+ F- and Al12B- Cs+. PMID- 17078600 TI - Intrinsic anion oxidation potentials. AB - Anions of lithium battery salts have been investigated by electronic structure calculations with the objective to find a computational measure to correlate with the observed (in)stability of nonaqueous lithium battery electrolytes vs oxidation often encountered in practice. Accurate prediction of intrinsic anion oxidation potentials is here made possible by computing the vertical free energy difference between anion and neutral radical (Delta Gv) and further strengthened by an empirical correction using only the anion volume as a parameter. The 6 311+G(2df,p) basis set, the VSXC functional, and the C-PCM SCRF algorithm were used. The Delta Gv calculations can be performed using any standard computational chemistry software. PMID- 17078601 TI - A simple multichromophore design for energy transfer in distyrylbenzenes with pyrene pendants. AB - A set of Don --> Acc <-- Don multichromophores was synthesized based on a simple design with two pyrene donors (absorbing antennae) connected to a bis-1,4-(3,4,5 trimethoxystyryl)benzene core acceptor using various flexible, nonconjugated tethers. Excitation of the pyrene donors at 276 nm in solution yields near exclusive emission from the core chromophore at 445-450 nm, with energy transfer efficiencies up to 92%, far better than achieved with simple mixtures. The simple tethering design imposes a high "local" concentration of the pyrene near the acceptor core unit that is maintained even at very low multichromophore concentrations. Solvent effects on absorption and emission spectra are very small, except in cases where a pi-conjugating O-C=O moiety of the tethering group is directly attached to the core chromophore, rather than being placed in the middle of the tether. Energy transfer in the systems is effective due to good donor-acceptor energy matching. The optimal energy transfer efficiency was achieved using an eight-atom flexible linker. PMID- 17078602 TI - Photoinduced intramolecular charge transfer reaction in (E)-3-(4-Methylamino phenyl)-acrylic acid methyl ester: A fluorescence study in combination with TDDFT calculation. AB - A donor-acceptor substituted aromatic system (E)-3-(4-Methylamino-phenyl)-acrylic acid methyl ester (MAPAME) has been synthesized, and its photophysical behavior obtained spectroscopically has been compared with the theoretical results. The observed dual fluorescence from MAPAME has been assigned to emission from locally excited and twisted intramolecular charge transfer states. The donor and acceptor angular dependency on the ground and excited states potential energy surfaces have been calculated both in vacuo and in acetonitrile solvent using time dependent density functional theory (TDDFT) and TDDFT polarized continuum model (TDDFT-PCM), respectively. Calculation predicts that a stabilized twisted excited state is responsible for red shifted charge transfer emission. PMID- 17078603 TI - Distribution of internal states of CO from O (1D) + CO determined with time resolved fourier transform spectroscopy. AB - Following collisions of O (1D) with CO, rotationally resolved emission spectra of CO (1 < or = v < or = 6) in the spectral region 1800-2350 cm(-1) were detected with a step-scan Fourier transform spectrometer. O (1D) was produced by photolysis of O3 with light from a KrF excimer laser at 248 nm. Upon irradiation of a flowing mixture of O3 (0.016 Torr) and CO (0.058 Torr), emission of CO (v < or = 6) increases with time, reaches a maximum approximately 10 micros. At the earliest applicable period (2-3 micros), the rotational distribution of CO is not Boltzmann; it may be approximately described with a bimodal distribution corresponding to temperatures approximately 8000 and approximately 500 K, with the proportion of these two components varying with the vibrational level. A short extrapolation from data in the period 2-6 micros leads to a nascent rotational temperature of approximately 10170 +/- 600 K for v = 1 and approximately 1400 +/- 40 K for v = 6, with an average rotational energy of 33 +/ 6 kJ mol(-1). Absorption by CO (v = 0) in the system interfered with population of low J levels of CO (v = 1). The observed vibrational distribution of (v = 2):(v = 3):(v = 4):(v = 5):(v = 6) = 1.00:0.64:0.51:0.32:0.16 corresponds to a vibrational temperature of 6850 +/- 750 K. An average vibrational energy of 40 +/ 4 kJ mol(-1) is derived based on the observed population of CO (2 < or = v < or = 6) and estimates of the population of CO (v = 0, 1, and 7) by extrapolation. The observed rotational distributions of CO (1 < or = v < or = 3) are consistent with results of previous experiments and trajectory calculations; data for CO (4 < or = v < or = 6) are new. PMID- 17078604 TI - Ultrafast relaxation dynamics of the excited states of Michler's thione. AB - Ultrafast relaxation dynamics of the S2 and S1 states of 4,4'-bis(N,N dimethylamino)thiobenzophenone (Michler's thione, MT) have been investigated in different kinds of solvents, using steady-state absorption and emission as well as femtosecond transient absorption and fluorescence up-conversion spectroscopic techniques. Steady-state fluorescence measurements, following photoexcitation to the S2 state of MT, reveal weak fluorescence from the S2 state (phi F approximately 10(-3) in nonpolar and 10(-4) in polar solvents) but much weaker fluorescence from the S1 state. Yield of fluorescence from the S2 state is reduced in polar solvents because of reduced energy gap between the S2 and S1 states, Delta E(S2-S1), as well as interaction with the solvent molecules. Occurrence of S2-fluorescence in polar solvents, despite small energy gap, suggests that symmetry allowed S2(1A1) --> S0 (1A1) radiative and symmetry forbidden S2(1A1) --> S1 (1A2) nonradiative transitions are the factors responsible for the S2 fluorescence in MT. Lifetime of the S2 state is shorter (varying in the range 0.28-3.5 ps in different solvents) than that predicted from the Delta E(S2-S1) value and this can be attributed to its flexible molecular structure, which promotes an efficient intramolecular radiationless deactivation pathways. The lifetime of the S1 state (approximately 1.9-6.5 ps) is also very short because of small energy difference between the S1 and T1 states (Delta E(S1 T1) approximately 300 cm(-1)) in cyclohexane and hydrogen-bonding interaction as well as the presence of the isoenergetic T1(pipi*) state to enhance the rate of the intersystem crossing process from the S1(npi*) state in protic solvents. PMID- 17078605 TI - Optical and X-ray emission spectroscopy of high-power laser-induced dielectric breakdown in molecular gases and their mixtures. AB - Large-scale plasma was created in molecular gases (CO, CO2, N2, H2O) and their mixtures by high-power laser-induced dielectric breakdown (LIDB). Compositions of the mixtures used are those suggested for the early earth's atmosphere of neutral and/or mildly reducing character. Time-integrated optical spectra emitted from the laser spark have been measured and analyzed. The spectra of the plasma generated in the CO-containing mixtures are dominated by emission of both C2 and CN radicals. A vibrational temperature of approximately 10(4) K was determined according to an intensity distribution in a vibronic structure of the CN (B2Sigma(+)u-X2Sigma(+)g) violet band. For comparison, the NH3-CH4-H2-H2O mixture has been irradiated as a model of the strongly reducing version of the early earth's atmosphere. In this mixture, excited CN seems to be significantly less abundant than C2. The LIDB experiments were in the molecular gases carried out not only in the static cell but also using a large, double stream pulse jet (gas puff target) placed in the vacuum interaction chamber. The obtained soft X-ray emission spectra indicate the presence of highly charged atomic ions in the hot core of high-power laser sparks. PMID- 17078606 TI - Nitrogen 1s near-edge X-ray absorption fine structure spectroscopy of amino acids: Resolving zwitterionic effects. AB - Considerable variation is observed in the near-edge X-ray absorption fine structure (NEXAFS) spectra of amino acids. To unambiguously characterize the chemical origin of this variation, we have acquired the nitrogen 1s NEXAFS spectra of several amino acids and other model compounds and complemented these experimental measurements with ab initio calculations of isolated molecules and molecular clusters. The systematic differences observed between the zwitterionic and un-ionized forms of amino acids arise directly from the structural difference (-NH2 vs -NH3+), which leads to a change in the degree of Rydberg-valence mixing. Further change arises from quenching of this Rydberg character in the spectra of condensed amino acids. Ab initio calculations are used to explore the degree of Rydberg-valence mixing in the solid state. PMID- 17078607 TI - Matrix-isolated van der Waals complexes formed between CS2 and dihalogen molecules XY, Where XY = Cl2, Br2, BrCl, ICl, or IBr. AB - Weakly bound 1:1 complexes formed between CS2 and a dihalogen molecule XY = Cl2, Br2, BrCl, ICl, or IBr have each been trapped in an Ar matrix and hence investigated experimentally by their IR spectra as well as theoretically by MP2 and density functional calculations. A planar structure, with an intermolecular angle close to 90 degrees , is expected for such a S=C=S...XY molecular complex. Moreover, for each system involving a heteronuclear dihalogen, two possible complexes exist, viz., S=C=S...XY and S=C=S...YX. The calculated structures, vibrational properties, and binding energetics of the complexes are analyzed, and the NBO formalism is used to interpret their bonding properties. The IR spectra of the complexes thus simulated provided vital guidance for the interpretation of the matrix spectra. For example, complexation was predicted and observed (i) to induce red shifts of the principal absorptions associated with both the CS2 and XY components and (ii) to result, through the change in symmetry, in activation of some modes that are IR-silent for the free components. PMID- 17078608 TI - Tuning excited-state electron transfer from an adiabatic to nonadiabatic type in donor-bridge-acceptor systems and the associated energy-transfer process. AB - Through design and synthesis of a new series of dyads I-III composed of 2,3 dimethoxynaphthalene as an electron donor (D) and 2,3-dicyanonaphthalene as an acceptor (A) bridged by n-norbornadiene (n = 1-3) we demonstrate an excellent prototype to switch the excited-state electron-transfer dynamics from an adiabatic to a nonadiabatic process. I reveals a remarkable excitonic effect and undergoes an adiabatic type of electron transfer (ET), resulting in a unique charge-transfer emission, of which the peak wavelength exhibits strong solvatochromism. Conversely, upon exciting the donor moiety, a fast D --> A energy transfer takes place for II (approximately 3 ps) and III (< or =30 ps), followed by a nonadiabatic type, weak coupled electron transfer with a relatively slow ET rate, giving rise to dual emission in polar solvents. Further detailed temperature-dependent studies of the ET rate deduced reaction barriers of 2.7 kcal/mol (for II) and 1.3 kcal/mol (for III) in diethyl ether and CH2Cl2, respectively. The results lead to a deduction of the reaction free energy and reorganization energy for both II (in diethyl ether) and III (in CH2Cl2). Theoretical (for I) and experimental (for II and III) approaches estimate the electronic coupling to be 860, 21.9, and 3.2 cm(-1) for I, II, and III, respectively, supporting the adiabatic versus nonadiabatic switching mechanism. PMID- 17078609 TI - Controlled excitations of the Belousov-Zhabotinsky reaction: Experimental procedures. AB - The purpose of this research was to explore the unstirred, ferroin-catalyzed Belousov-Zhabotinsky (BZ) reaction as an experimental model for the response of excitable media to small perturbations (slightly larger than the threshold for excitations). Following Showalter et al. (Showalter, K.; Noyes, R. M.; Turner, H. J.Am. Chem. Soc. 1979, 101, 7463-69), we used a positively biased silver electrode to release silver ions into a BZ reaction mixture, removing bromide ions and causing an excitation if sufficient bromide was removed. We found (1) a scaling region in which the delay before activation increased linearly as the size of the perturbation decreased, qualitatively consistent with but not fully explained by the Oregonator of Field et al. (Field, R. J.; Koros, E.; Noyes, R. M. J. Am. Chem. Soc. 1972, 94, 8649-64); (2) evidence for a 10 s oligomerization time scale; and (3) that activations were always delayed until after the end of a pulse of current, with the delay essentially constant for sufficiently long pulses, an effect not seen in simple ODE models but consistent with the anomalously large current apparently required for activation (Showalter, K.; Noyes, R. M. J. Am. Chem. Soc. 1976, 98, 3730-31) and explainable by bromide transport. Overall, the BZ system appeared to be well-suited as an experimental prototype, despite its complexity. PMID- 17078610 TI - Kinetics study of OH radical reactions with n-octane, n-nonane, and n-decane at 240-340 K using the relative rate/discharge flow/mass spectrometry technique. AB - The kinetics of the reactions of hydroxyl radical with n-octane (k1), n-nonane (k2), and n-decane (k3) at 240-340 K and a total pressure of approximately 1 Torr has been studied using relative rate combined with discharge flow and mass spectrometer (RR/DF/MS) technique. The rate constant for these reactions was found to be positively dependent on temperature, with an Arrhenius expression of k1 = (2.27 +/- 0.21) x 10(-11)exp[(-296 +/- 27)/T], k2 = (4.35 +/- 0.49) x 10( 11)exp[(-411 +/- 32)/T], and k3 = (2.26 +/- 0.28) x 10(-11)exp[(-160 +/- 36)/T] cm3 molecule(-1) s(-1) (uncertainties taken as 2sigma), respectively. Our results are in good agreement with previous studies at and above room temperature using different techniques. Assuming that the reaction of alkane with hydroxyl radical is the predominant form for loss of these alkanes in the troposphere, the atmospheric lifetime for n-octane, n-nonane, and n-decane is estimated to be about 43, 35, and 28 h, respectively. PMID- 17078611 TI - Equilibrium phase diagrams of aqueous mixtures of malonic acid and sulfate/ammonium salts. AB - Tropospheric aerosols are usually complex mixtures of inorganic and organic components. Although the thermodynamic properties of inorganic aerosols have been widely studied, the effect of organics on such properties is still under discussion. In this study, solubility in water, water activity (a(w)) of aqueous solutions, deliquescence relative humidity (DRH), eutonic composition, and eutonic DRH were determined for bulk mixtures of malonic acid (MA) with ammonium sulfate (AS) and ammonium bisulfate (ABS) at 25 degrees C over the full range of composition (from 0 wt % to the solubility limit of the mixture components). The data were used to construct equilibrium phase diagrams, which show the phase of the mixtures as a function of total composition, dry mixture composition, water content, and ambient relative humidity (RH). This work complements previous reports on the thermodynamic properties of AS/MA mixtures because the range of concentrations investigated is larger than in any other published single study. On the other hand, this is the first report on the a(w), deliquescence, and water absorption of ABS/MA mixtures. The eutonic composition for AS/MA mixtures was found to be 66.8 MA dry wt % (MA dry wt % = MA mass x 100/(AS mass + MA mass) with a DRH of 0.437. The eutonic composition for the ABS/MA mixtures was lower than for the AS/MA mixtures: 20.9 MA dry wt % with a DRH of 0.327. Measured a(w) of liquid AS/MA and ABS/MA solutions is compared with an extended Zdanovskii Stokes-Robinson expression, obtaining a good agreement (error < 5-6%). The expression was used to predict water uptake of mixtures and might be useful to interpret particle hygroscopic growth experiments. Comparison of the AS/MA and ABS/MA systems indicates that ABS reduces the DRH and enhances water uptake, relative to mixtures with AS. The results confirm that ambient particles containing sulfate and water-soluble organic compounds can remain liquid or partially liquid at very low ambient RH conditions, especially if the sulfate is not completely neutralized. PMID- 17078612 TI - Quantum chemical and statistical rate study of the reaction of O(3P) with allene: O-addition and H-abstraction channels. AB - The lowest-lying triplet and singlet potential energy surfaces for the O(3P) + CH2=C=CH2 reaction were theoretically characterized using the complete basis set model chemistry, CBS-QB3. The primary product distributions for the multistate multiwell reactions on the individual surfaces were then determined by RRKM statistical rate theory and weak-collision master equation analysis using the exact stochastic simulation method. The results predict that the electrophilic O addition pathways on the central and terminal carbon atom are dominant up to combustion temperatures. Major predicted end-products for the addition routes include CO + C2H4, 3CH2 + H2CCO, and CH2=C*-CHO + H*, in agreement with experimental evidence. CO + C2H4 are mainly generated from the lowest-lying singlet surface after an intersystem crossing process from the initial triplet surface. Efficient H-abstraction pathways are newly identified and occur on two different electronic state surfaces, 3A'' and 3A', resulting in OH + propargyl radicals; they are predicted to play an important role at higher temperatures in hydrocarbon combustion chemistry and flames, with estimated contributions of ca. 35% at 2000 K. The overall thermal rate coefficient k(O + C3H4) at 200-1000 K was computed using multistate transition state theory: k(T) = 1.60 x 10(-17) x T (2.05) x exp(-90 K/T) cm3 molecule(-1) s(-1), in good agreement with experimental data available for the 300-600 K range. PMID- 17078613 TI - Kinetics of the reactions of CH2I, CH2Br, and CHBrCl radicals with NO2 in the temperature range 220-360 K. AB - The kinetics of the CH2I + NO2, CH2Br + NO2, and CHBrCl + NO2 reactions have been studied at temperatures between 220 and 360 K using laser photolysis/photoionization mass spectrometry. Decays of radical concentrations have been monitored in time-resolved measurements to obtain reaction rate coefficients under pseudo-first-order conditions. The bimolecular rate coefficients of all three reactions are independent of the bath gas (He or N2) and pressure within the experimental range (2-6 Torr) and are found to depend on temperature as follows: k(CH2I + NO2) = (2.18 +/- 0.07) x 10(-11) (T / 300 K)( 1.45) (+/- 0.22) cm3 molecule(-1) s(-1) (220-363 K), k(CH2Br + NO2) = (1.76 +/- 0.03) x 10(-11) (T/300 K)(-0.86) (+/- 0.09) cm3 molecule(-1) s(-1) (221-363 K), and k(CHBrCl + NO2) = (8.81 +/- 0.28) x 10(-12) (T/300 K)(-1.55) (+/- 0.34) cm3 molecule(-1) s(-1) (267-363 K), with the uncertainties given as one-standard deviations. Estimated overall uncertainties in the measured bimolecular reaction rate coefficients are about +/-25%. In the CH2I + NO2 and CH2Br + NO2 reactions, the observed product is formaldehyde. For the CHBrCl + NO2 reaction, the product observed is CHClO. In addition, I atom and iodonitromethane (CH2INO2) or iodomethyl nitrite (CH2IONO) formations have been detected for the CH2I + NO2 reaction. PMID- 17078614 TI - Kinetics of the CN + HCNO reaction. AB - The kinetics of the CN + HCNO reaction were studied using laser-induced fluorescence and infrared diode laser absorption spectroscopy. The total rate constant was measured to be k(T) = (3.95 +/- 0.53) x 10(-11) exp[(287.1 +/- 44.5)/T] cm3 molec(-1) s(-1), over the temperature range 298-388 K, with a value of k1 = (1.04 +/- 0.1) x 10(-10) cm3 molec(-1) s(-1) at 298 K. After detection of products and consideration of secondary chemistry, we conclude that NO + HCCN is the only major product channel. PMID- 17078615 TI - Comparison of the localization of an electron as determined by the two-particle distribution function and by the single-particle sharing index. AB - A comparison of the measure of the delocalization of a particle based on the two particle distribution function and that based on the single-particle density matrix is made using a simple set of wave functions which span states ranging from single determinant ground and doubly excited states through states mimicking correlated states and which include the singly excited state for electrons and for bosons replacing electrons in H2. The comparison further includes an analysis of the application of the measures to a classical ideal gas and a compressible fluid. It is found that the values of the integrated atom-atom measures agree for a range of wave functions involving combinations of the two single determinant (and equivalent Bose) wave functions but disagree for a different range of these wave functions and for the singly excited wave functions. Aside from the single determinant (and equivalent Bose) wave functions, the two sets of point-point measures that underlie the integrated measures all differ. For the sets of wave functions considered, the values of the measures are identical for electrons and bosons. When applied to a closed classical ideal gas and to a closed compressible fluid, the delocalization measure based on the two-particle distribution has a residual long range term, whereas the sharing index in the classical limit gives a completely localized particle. In general, the two measures describe different aspects of the behavior of the particles. The measures based on the two-particle distribution function give only two-particle properties and the single-particle density, and the sharing quantities give only single-particle properties. The latter includes, however, the quantitative measures of the delocalization of a single particle, the point-point sharing index and the sharing amplitude. PMID- 17078616 TI - Theoretical investigation of linalool oxidation. AB - This study concerns the autoxidation of one of the most used fragrances in daily life, linalool (3,7-dimethyl-1,6-octadien-3-ol). It reacts with O2 to form hydroperoxides, which are known to be important contact allergens. Pathways for hydroperoxide formation are investigated by means of quantum mechanical electronic structure calculations. Optimized molecular geometries and harmonic vibrational frequencies are determined using density functional theory (DFT). Insight into how the addition of O2 to linalool occurs is obtained by establishing a theoretical framework and systematically investigating three smaller systems: propene, 2-methyl-2-butene, and 2-methyl-2-pentene. 2-Methyl-2 pentene was chosen as a model system and used to compare with linalool. This theoretical study characterizes the linalool-O2 biradical intermediate state, which constitutes a branching point for the further oxidation reactions pathways. Thus, the observed linalool oxidation product spectrum is discussed in terms of a direct reaction path, the ene-type mechanism, and the radical mechanism. The major hydroperoxide found in experiments is 7-hydroperoxy-3,7-dimethyl-octa-1,5 diene-3-ol, and the calculated results support this finding. PMID- 17078617 TI - DFT and TDDFT study related to electron transfer in nonbonded porphine...C60 complexes. AB - Spectroscopic properties of a ground state nonbonded porphine buckminsterfullerene (H2P...C60) complex are studied in several different relative orientations of C60 with respect to the porphine plane by using the density functional (DFT) and time-dependent density functional (TDDFT) theories. The geometries and electronic structures of the ground states are optimized with the B3LYP and PBE functionals and a SVP basis set. Excitation energies and oscillator strengths are obtained from the TDDFT calculations. The relative orientation of C60 is found to affect the equilibrium distance between H2P and C60 especially in the case of the PBE functional. The excitation energies of different H2P...C60 complexes are found to be practically the same for the same excitations when the B3LYP functional is used but to differ notably when PBE is used in calculations. Existence of the states related to a photoinduced electron transfer within a porphyrin-fullerene dyad is also studied. All calculations predict a formation of an excited charge-transfer complex state, a locally excited donor (porphine) state, as well as a locally excited acceptor (fullerene) state in the investigated H2P...C60 complexes. PMID- 17078618 TI - Theoretical study of two-photon absorption properties of a series of ferrocene based chromophores. AB - The electronic structures, one-photon absorption (OPA), and two-photon absorption (TPA) properties of a series of ferrocene-based chromophores with TCF-type acceptors (TCF = 2-dicyanomethylene-3-cyano-4-methyl-2,5-dihydrofuran) have been studied by using the ZINDO-SOS method. The results have revealed that OPA and TPA of ferrocenyl derivatives are affected by the strength of the acceptor, especially the pi-bridge conjugation length. The TPA cross section increases with increasing acceptor strength and pi-bridge conjugation length. The TCF-type acceptor with a phenyl group can lead to a larger TPA cross section. Quadrupole molecules have the largest TPA cross sections (2000-3000 GM), which are about 4 times that of the corresponding dipolar molecules, indicating larger interactions between the top and bottom branches. Finally, the origins of the two-photon excitations for ferrocenyl derivatives are analyzed. The calculations show that ferrocenyl derivatives with TCF-type acceptors (especially quadrupole molecules) are promising candidates for TPA materials. PMID- 17078619 TI - Electronic structure analysis and electron detachment energies of polynitrogen pentagonal aromatic anions. AB - Various decouplings of the electron propagator have been employed to provide theoretical comparison to experimental electron detachment energies for the pyrrolide, imidazolide, and pyrazolide anions. Predictions for isoelectronic anions in which CH groups are replaced by N atoms also are reported. The ab initio electron propagator results agree closely with experimental values, and the associated Dyson orbitals provide a detailed catalog of bonding changes as the number and positions of N atoms vary within the set of pentagonal aromatic anions. PMID- 17078620 TI - Induction of an aromatic six-membered nitrogen ring via cation-pi interaction. AB - Nitrogen clusters have been intensively studied for their potential application as high-energy density materials, but a six-membered nitrogen ring (N6) was not found to be stable and aromatic. To explore the possibility of inducing an aromatic N6 ring via cation-pi interaction, quantum chemistry calculations were performed on the systems of Ca2N6, CaN6, CaN6(2-), N6, and N6(4-) at the B3LYP/6 311+G level. The optimized geometries reveal that the planar structure of the N6 ring is stable only in the Ca2N6 complex. The computed NBO and CHelpG charges demonstrate that the planar N6 moiety in the Ca2N6 complex is almost a 10pi electron system. The predicted nucleus-independent chemical shift (NICS) values demonstrate that the N6 moiety is aromatic in comparison with the NICS values of benzene. The estimated enthalpy of formation for the Ca2N6 complex is 100.4 kcal/mol for the reaction of 2Ca and 3N2. The binding energy between the Ca2+ cation and the N6(4-) moiety is -1928.8 kcal/mol, with electrostatic interaction serving as the predominant component. When all the calculated results are taken into account, including the planar structure, 10pi-electron system, identical bond length, and negative NICS value of the N6(4-) moiety in the Ca2N6 complex, it is deduced that the alkaline earth metal Ca is capable of inducing an aromatic N6 ring through the cation-pi interaction formed by electron transfer from the Ca atom to the N6 ring. PMID- 17078621 TI - Density functional study of chemical stability and nitrogen encapsulation of C48N12 and C58N12. AB - On the basis of calculations using density functional theory, we show that C58N12, just as C48N12, can be a stable N-dopant of C70. By considering many different isomers of the product, we find that the chemical stability of C48N12 and C58N12, with respect to oxygenation, is not significantly different from that of C70, thereby indicating that the N-dopant would not easily be oxygenated in air under normal conditions. In both C48N12O and C58N12O, many different isomers are expected, in which oxygenation occurs at different C-N bonds as well as at C C bonds, among which specific C-N bonds are the most amenable to the reaction. Investigation of their hydrogenations shows that C48N12 is slightly more easily hydrogenated than C60, while C58N12 is less easily hydrogenated. In addition, we expect a regiospecificity in the hydrogenated products of C58N12, which prefers to react at equatorial sites, while C70 prefers reaction at polar sites. Meanwhile, comparison of the encapsulation energy of a nitrogen atom (=N en) in C60, C48N12, C70, and C58N12 shows that the N-doped fullerenes, particularly C58N12, can encase the atom much better than the undoped ones, allowing us to expect the existence of N@C48N12 and N@C58N12. Spin multiplicities are doublet for most of their stable structures. These observations correlate with the formation of N en-C bonds, which are not found in N@C60 and N@C70. Various isomers of the N-encapsulating fullerenes were identified. The relative stability of these isomers heavily depends on the number of substitutional nitrogen atoms around N en-C bonds. PMID- 17078622 TI - Local aromaticity in natural nucleobases and their size-expanded benzo-fused derivatives. AB - The influence of the insertion/addition of a benzene ring to the natural nucleic acid bases on the local aromaticity of the so-called size-expanded (xN, with N being adenine, guanine, cytosine, and thymine) bases is examined. To this end, the local aromaticity of the six- and five-membered rings in both the natural bases and their benzoderivatives is determined using HOMA, NICS, aromatic fluctuation index (FLU), and para-delocalization index (PDI) descriptors. In general, there is a good correspondence between the different indices, so that ring moieties with more negative NICS values also have larger HOMA and PDI measures and lower FLU indices. The results also point out notable differences in the aromatic character of the natural and size-expanded bases, which generally are hardly affected upon hydrogen bonding. The differences in the highest occupied molecular orbital-lowest unoccupied molecular orbital (HOMO-LUMO) gap determined for the size-expanded nucleobases show an inverse correlation with the aromaticity of the fused benzene ring, so that the larger the HOMO-LUMO gap is, the lower the destabilization experienced by the benzene upon insertion/addition to the natural bases. This finding suggests that the introduction of suitable chemical modifications in the benzene ring might be useful to modulate the HOMO LUMO gap while enabling the design of modified DNA duplexes that are able to act as molecular wires. PMID- 17078623 TI - Bond length and local energy density property connections for non-transition metal oxide-bonded interactions. AB - For a variety of molecules and earth materials, the theoretical local kinetic energy density, G(r(c)), increases and the local potential energy density, V(r(c)), decreases as the M-O bond lengths (M = first- and second-row metal atoms bonded to O) decrease and the electron density, rho(r(c)), accumulates at the bond critical points, r(c). Despite the claim that the local kinetic energy density per electronic charge, G(r(c))/rho(r(c)), classifies bonded interactions as shared interactions when less than unity and closed-shell when greater, the ratio was found to increase from 0.5 to 2.5 au as the local electronic energy density, H(r(c)) = G(r(c)) + V(r(c)), decreases and becomes progressively more negative. The ratio appears to be a measure of the character of a given M-O bonded interaction, the greater the ratio, the larger the value of rho(r(c)), the smaller the coordination number of the M atom and the more shared the bonded interaction. H(r(c))/rho(r(c)) versus G(r(c))/rho(r(c)) scatter diagrams categorize the M-O bonded interactions into domains with the local electronic energy density per electron charge, H(r(c))/rho(r(c)), tending to decrease as the electronegativity differences for the bonded pairs of atoms decrease. The values of G(r(c)) and V(r(c)), estimated with a gradient-corrected electron gas theory expression and the local virial theorem, are in good agreement with theoretical values, particularly for the bonded interactions involving second-row M atoms. The agreement is poorer for shared C-O and N-O bonded interactions. PMID- 17078625 TI - Ab initio study on one-way photoisomerization of the maleic acid and fumaric acid anion radical system as a model system of their esters. AB - Potential energy surfaces (PESs) of the maleic acid anion radical (MA-*: cis isomer)/fumaric acid anion radical (FA-*: trans isomer) system as a model system of their esters have been studied in detail using CASSCF method. The results suggest the following: The photoisomerization is initiated with the H-C-C-H dihedral angle distortion [hydrogen out of plain (HOOP) motion] on the D1 PES. The C-C-C-C dihedral angle distortion occurs on the D0 PES after the deactivation from D1 to D0. A large fraction of the net motion along the isomerization coordinate occurs on the D0 PES. The D0 state is responsible for the one-way nature of the photoisomerization. PMID- 17078624 TI - Spin-spin contributions to the zero-field splitting tensor in organic triplets, carbenes and biradicals-a density functional and ab initio study. AB - An evaluation study for the direct dipolar electron spin-spin (SS) contribution to the zero-field splitting (ZFS) tensor in electron paramagnetic resonance (EPR) spectroscopy is presented. Calculations were performed on a wide variety of organic systems where the SS contribution to the ZFS dominates over the second order spin-orbit coupling (SOC) contribution. Calculations were performed using (hybrid) density functional theory (DFT), as well as complete active space self consistent field (CASSCF) wave functions. In the former case, our implementation is an approximation, because we use the two-particle reduced spin-density matrix of the noninteracting reference system. In the latter case, the SS contribution is approximated by a mean-field method which, nevertheless, gives accurate results, compared to the approximation free computation of the SS part in a CASSCF framework. For the case of the triplet dioxygen molecule, it was shown that restricted open-shell density functional theory (RODFT), as well as CASSCF, can provide accurate spin-spin couplings while spin-unrestricted DFT leads to much larger errors. Furthermore, 15 organic radicals, including several 1,3 and 1,5 diradicals, dinitroxide biradicals, and even a chlorophyll a model system, were examined as test cases to demonstrate the accuracy and efficiency of our approach within a DFT framework. Accurate D values with root-mean-square deviations of 0.0035 cm(-1) were obtained. Furthermore, all trends, including those due to substituent effects, were correctly reproduced. In a different set of calculations, the polyacenes benzene, naphthalene, anthracene, and tetracene were studied. Applying DFT, the absolute D values were noticeably underestimated, but it was possible to correctly reproduce the trend to smaller D values with larger size of the systems. Finally, it was demonstrated that our approach is also well-suited for the study of carbenes. The smaller organic radicals of this work were also studied, through the use of CASSCF wave functions. This was a special advantage in the case of the triplet polyacenes, where the CASSCF approach gave better results than the DFT method. In comparing spin-restricted and spin-unrestricted results, it was shown through a natural orbital analysis and comparison to high-level ab initio calculations that even small amounts of spin polarization introduced by the unrestricted calculations lead to large deviations between the unrestricted Kohn-Sham (UKS) and restricted open-shell Kohn-Sham (ROKS) approaches. It is challenging to understand why the ROKS results show much better correlation with the experimental data. PMID- 17078626 TI - Gaseous arginine conformers and their unique intramolecular interactions. AB - Extensive ab initio calculations were employed to characterize stable conformers of gaseous arginine, both the canonical and zwitterionic tautomers. Step-by-step geometry optimizations of possible single-bond rotamers at the B3LYP/6-31G(d), B3LYP/6-31++G(d,p), and MP2/6-31++G(d,p) levels yield numerous structures that are more stable than any known ones. The final electronic energies of the conformers were determined at the CCSD/6-31++G(d,p) level. The lowest energies of the canonical and zwitterionic structures are lower than the existing values by 2.0 and 2.3 kcal/mol, respectively. The relative energies, rotational constants, dipole moments, and harmonic frequencies of the stable conformers remain for future experimental verification. The conformational distributions at various temperatures, estimated according to thermodynamic principles, consist almost exclusively of the newly found structures. One striking feature is the occurrence of blue-shifting hydrogen bonds in all six of the most stable conformers. A unique feature of important conformations is the coexistence of dihydrogen and blue- and red-shifting hydrogen bonds. In addition to the hydrogen bonds, the stereoelectronic effects were also found to be important stabilization factors. The calculated and measured proton affinities agree within the theoretical and experimental uncertainties, affirming the high quality of our conformational search. The theoretical gas-phase basicity of 245.9 kcal/mol is also in good agreement with the experimental value of 240.6 kcal/mol. The extensive searches establish firmly that gaseous arginine exists primarily in the canonical and not the zwitterionic form. PMID- 17078627 TI - Spectroscopic and computational studies of the intramolecular hydrogen bonding of 2-indanol. AB - 2-Indanol in its most stable form is stabilized by internal hydrogen bonding, which exists between the hydroxyl hydrogen atom and the pi-cloud of the benzene ring. A comprehensive ab initio calculation using the MP2/cc-pVTZ level of theory showed that 2-indanol can exist in four possible conformations, which can interchange through the ring-puckering vibration and the internal rotation of the OH group on the five-membered ring. A potential energy surface in terms of these two vibrational coordinates was calculated. Density functional theory calculations were used to predict the vibrational frequencies and to help in normal mode assignments. Fluorescence excitation spectra of 2-indanol confirm the presence of the four conformers in the electronic ground and excited states. The spectral intensities indicate that, at 90 degrees C, 82% of the molecules exist in its most stable form with the intramolecular hydrogen bonding. The other isomers are present at approximately 11, 5, and 3%. The MP2/6-311++G(d,p) calculation predicts a distribution of 70, 13, 9, and 8% at 90 degrees C, the experimental sample temperature. PMID- 17078628 TI - Molecular electrostatic potential devices on graphite and silicon surfaces. AB - We demonstrate that molecular gates using molecular electrostatic potentials (MEP) can be used on hydrogen-passivated silicon substrates without any disturbance of their behavior in vacuum; however, the use of graphite as a substrate strongly affects such behavior. As expected, the substrate may become one more design variable. The ability to have several substrate alternatives is very important for the practical implementation of this new scenario based on molecular potentials. In general, the effect of the substrate can be predetermined by calculating the MEP of the surface as this indicates how strongly its intrinsic potential is. PMID- 17078629 TI - Electronic states and spectroscopic properties of SiTe and SiTe+. AB - Ab initio based configuration interaction calculations have been carried out to study the low-lying electronic states and spectroscopic properties of the heaviest nonradioactive silicon chalcogenide molecule and its monopositive ion. Spectroscopic constants and potential energy curves of states of both SiTe and SiTe+ within 5 eV are reported. The calculated dissociation energies of SiTe and SiTe+ are 4.41 and 3.52 eV, respectively. Effects of the spin-orbit coupling on the electronic spectrum of both the species are studied in detail. The spin-orbit splitting between the two components of the ground state of SiTe+ is estimated to be 1880 cm(-1). Transitions such as 0+ (II)-X1Sigma(+)0+, 0+ (III)-X1Sigma(+)0+, E1Sigma(+)0+ -X1Sigma(+)0+, and A1Pi1-X1Sigma(+)0+ are predicted to be strong in SiTe. The radiative lifetime of the A1Pi state is less than a microsecond. The X(2)2Pi(1/2)-X(1)2Pi(3/2) transition in SiTe+ is allowed due to spin-orbit mixing. However, it is weak in intensity with a partial lifetime for the X2 state of about 108 ms. The electric dipole moments of both SiTe and SiTe+ in their low lying states are calculated. The vertical ionization energies for the ionization of the ground-state SiTe to different ionic states are also reported. PMID- 17078630 TI - DFT/B3LYP study of the substituent effect on the reaction enthalpies of the individual steps of single electron transfer-proton transfer and sequential proton loss electron transfer mechanisms of phenols antioxidant action. AB - The reaction enthalpies related to the individual steps of two phenolic antioxidants action mechanisms, single electron transfer-proton transfer (SET-PT) and sequential proton loss electron transfer (SPLET), for 30 meta and para substituted phenols (ArOH) were calculated using DFT/B3LYP method. These mechanisms represent the alternative ways to the extensively studied hydrogen atom transfer (HAT) mechanism. Except the comparison of calculated reaction enthalpies with available experimental and/or theoretical values, obtained enthalpies were correlated with Hammett constants. We have found that electron donating substituents induce the rise in the enthalpy of proton dissociation (PDE) from ArOH+* radical cation (second step in SET-PT) and in the proton affinities of phenoxide ions ArO- (reaction enthalpy of the first step in SPLET). Electron-withdrawing groups cause the increase in the reaction enthalpies of the processes where electron is abstracted, i.e., in the ionization potentials of ArOH (first step in SET-PT) and in the enthalpy of electron transfer from ArO- (second step in SPLET). Found results indicate that all dependences of reaction enthalpies on Hammett constants of the substituents are linear. The calculations of liquid-phase reaction enthalpies for several para-substituted phenols indicate that found trends hold also in water, although substituent effects are weaker. From the thermodynamic point of view, entering SPLET mechanism represents the most probable process in water. PMID- 17078631 TI - Chemically accurate thermochemistry of cadmium: An ab initio study of Cd + XY (X = H, O, Cl, Br; Y = Cl, Br). AB - Using a composite coupled cluster method employing sequences of correlation consistent basis sets for complete basis set (CBS) extrapolations and with explicit treatment of core-valence correlation and scalar and spin-orbit relativistic effects, the 0 K enthalpies of a wide range of cadmium-halide reactions, namely, Cd + (HCl, HBr, ClO, BrO, Cl2, BrCl, Br2) have been determined to an estimated accuracy of +/-1 kcal/mol. In addition, accurate equilibrium geometries, harmonic frequencies, and dissociation energies have been calculated at the same level of theory for all the diatomic (e.g., CdH, CdO, CdCl, CdBr) and triatomic (CdHCl, CdHBr, CdClO, CdBrO, CdCl2, CdBrCl, CdBr2) species involved in these reactions, some for the very first time. Like their mercury analogues, all of the abstraction reactions are predicted to be endothermic, while the insertion reactions are strongly exothermic with the formation of stable linear, Cd-centric complexes. With the exception of CdH and the reactions involving this species, the present results for the remaining Cd-containing systems are believed to be the most accurate to date. PMID- 17078632 TI - Computer-aided design of promising photochemical alkoxy radical precursors. AB - A computer-aided design of alkoxyl radical precursors is performed. The new precursors should combine the advantages of N-alkoxypyridine-2(1H)thiones (less reactive radicals) and N-alkoxythiazole-2(3H)thiones (stable with respect to daylight). Additionally, the radical liberation process should be initiated by light with a wavelength of around 350 nm. To find promising compounds, 18 test candidates were obtained by a systematic variation of the parent compound N alkoxythiazole-2(3H)thione. The properties of the test molecules were computed by a protocol that was already successfully used to rationalize the photochemical behavior of N-alkoxypyridine-2(1H)thiones and N-alkoxythiazole-2(3H)thiones. The computations identify two promising new compounds. For N-methoxy-(1,3)dihydro [1,3]azaphosphole-2-thione (6a), they predict that the fragmentation process will be initiated by an absorption at 348 nm. An analysis of its fragmentation process indicates that the free excess energy of the resulting radicals should more resemble the situation found for N-alkoxypyridine-2(1H)thiones. For N-methoxy (1,3)dihydro-pyrrole-2-thione (3a), the excitation energy is somewhat higher (330 nm), but the computed fragmentation paths again indicate that the remaining excess energy of the released radicals is quite favorable. The test molecules also contained the experimentally well-known N-methoxypyridine-2(1H)one (1b). For this molecule, our computed data rationalizes nicely the experimental findings. PMID- 17078633 TI - Density functional calculations of 3He chemical shift in endohedral helium fullerenes: Neutral, anionic, and di-helium species. AB - We report density functional calculations of 3He nuclear magnetic resonance chemical shifts in a series of experimentally known endohedral helium fullerenes, He(n)@Cm(q) (n = 1, 2; m = 60, 70, 76, 78; q = 0, 6-), including for the first time anionic and di-helium species. Despite the lack of dispersion in the density functional model, the results are in promising agreement with experiment. Density functional theory performs better than Hartree-Fock for the anionic systems. In the di-helium species confined in the small C60 cage, besides the atomic displacements from the center position, the direct He-He interactions contribute to the 3He shift. PMID- 17078634 TI - Very large infrared two-photon absorption cross section of asymmetric zinc porphyrin aggregates: Role of intermolecular interaction and donor-acceptor strengths. AB - Very large two-photon absorption (TPA) cross sections at the infrared region have been revealed for J-aggregates of asymmetric zinc porphyrin using quantum chemical calculation. The TPA properties are evaluated for monomer and aggregates of a series of push-pull porphyrins, whose syntheses are known in the literature. The two-photon absorption cross section can be greatly enhanced by increasing the strengths of the electron donor/acceptor. We also present a quantum-chemical analysis on porphyrin aggregates to understand the role of intermolecular interactions and the relationship between structural and collective nonlinear optical properties. It has been observed that the TPA properties change tremendously as monomers undergo J-aggregation and the magnitudes of TPA cross sections are highly dependent on the nature of aggregates. The importance of our results with respect to the design of photonic and photodynamic therapy materials has been discussed. PMID- 17078635 TI - Quantitative structure-activity (affinity) relationship (QSAR) study on protonation and cationization of alpha-amino acids. AB - A quantitative structure-activity (affinity) relationship (QSAR) study is carried out to model the proton, sodium, copper, and silver cation affinities of alpha amino acids (AA). Stepping multiple linear regression (MLR), partial least squares (PLS), and artificial neural network (ANN) approaches are applied to elucidate the multiple factors affecting these affinities. The MLR and PLS models reveal that the variation in proton affinity is attributed to the highest electrophilic superdelocalizability of nitrogen (major) and the number of rotatable bonds (minor) in AA. The noncovalent interactions, especially ion dipole interactions, are responsible for the changes in Na+ affinity. The ionization potential, dipole moment of the side chain, and degree of linearity are the properties of AA that give the best correlation with the Cu+ and Ag+ affinities. The ANN models are developed to study the relationships (linear or nonlinear) between the molecular descriptors and binding affinities. The ANN models show higher predictive power. The QSAR models are used to study the binding forms of AA (neutral vs zwitterionic) upon protonation/cationization. To our knowledge, this is the first attempt to carry out a QSAR study on protonated/cationized AlphaAlpha to elucidate their binding properties. In virtue of the Na+ affinity ANN model, the Na+ affinities of dihydroxyphenylalanine (DOPA) were predicted. This work may pave the way for the success of applying similar approaches to peptides or proteins (with AA as the building blocks) in the future. PMID- 17078637 TI - Charles M. Knobler Festschrift comments. PMID- 17078640 TI - Bending and radial deformation of lipid tubules on self-assembled thiol monolayers. AB - Lipid tubules represent a hollow, cylindrical supramolecular structure formed by rolled-up lipid bilayers. We find that the lipid tubules of 1,2-bis(tricosa-10,12 diynoyl)-sn-glycero-3-phosphocholine can be bent into a loopike shape by the shrinking contact line of droplets on self-assembled monolayers (SAMs) of 1 dodecanethiol. The persistence length of individual lipid tubules is estimated to be approximately 41 microm. The radial deformation of the lipid tubules on SAMs is studied under applied load using atomic force microscope. The stiffness of the tubules in the radial direction is found to increase when the number of the lipid bilayers in the tubule wall increases. PMID- 17078641 TI - Distribution of coexisting solid and fluid phases alters the kinetics of collapse from phospholipid monolayers. AB - To determine how coexistence of liquid-expanded (LE) and tilted-condensed (TC) phases in phospholipid monolayers affects collapse from the air/water interface, we studied binary films containing dioleoyl phosphatidylcholine-dipalmitoyl phosphatidylcholine (DPPC) mixtures between 10 and 100% DPPC. Previously published results established that this range of compositions represents the LE TC coexistence region at the equilibrium spreading pressure of 47 mN/m. When held at 49.5 mN/m on a captive bubble, the extent of total collapse fit with the LE area predicted by the phase diagram. The kinetics of collapse, however, when normalized for changes in the LE area, slowed with increasing mole fraction of DPPC. Surface area expressed as stretched exponential functions of time yielded an Avrami exponent that decreased from 1 for the homogeneously LE film to 0.3 for DPPC > or = 70%. Microscopic studies showed that the largest changes in kinetics occurred when either alterations of the initial composition or the process of collapse induced the films to cross the percolation threshold, so that the LE phase became divided into isolated domains. Our results show that although coexisting solid and fluid phases collapse to extents that are independent, the kinetics of collapse, corrected for differences in LE area, depend on the distribution of the two phases. PMID- 17078642 TI - Contrast enhancement by feedback fields in magnetic resonance imaging. AB - A conceptually new approach giving rise to contrast enhancement by feedback fields in magnetic resonance imaging is proposed, and the detailed mechanism is described. Nonlinear spin dynamics under the feedback fields of the distant dipolar field and/or radiation damping are examined and shown to amplify contrast due to small variations in spin density and precession frequency. Feedback-based contrast enhancement depends on the instability of the initial magnetization configuration and is propagated by positive feedback, as shown through numerical simulations and experimental results on simple phantom samples. On the basis of a theoretical understanding of contrast enhancement, insight into pulse sequence design and optimal contrast attainable under the individual and joint feedback fields is provided. PMID- 17078643 TI - Headgroup percolation and collapse of condensed langmuir monolayers. AB - We present a study of Langmuir isotherms and 2D bulk moduli of binary lipid mixtures, where changes in monolayer collapse pressure (Pic) are followed while varying the relative amounts of the two components. For monolayers containing dipalmitoylphosphocholine (DPPC) with either hexadecanol (HD) or palmitic acid (PA), a distinctly non-monotonic change in Pic is observed with varying composition. At low mole fractions, there is a slight decrease in Pic as films get richer in DPPC, while a sharp increase to pure DPPC-like values is observed when the mole fraction exceeds approximately 0.7. The sudden transition in collapse pressure is explained using the principles of rigidity percolation, and important ramifications of this phenomenon for biological surfactant are discussed. PMID- 17078644 TI - Synthesis and self-assembly of an amphiphilic poly(phenylene ethynylene) ionomer. AB - We have synthesized a conjugated amphiphilic polyelectrolyte, a poly(phenylene ethynylene) (PPE), and the structurally analogous neutral polymer. The solution phase aggregation of the uncharged PPE can be reversibly controlled by varying the solvent polarity and concentration, while the charged polymer appears to self assemble at any concentration in compatible solvents. These conclusions are based on a combination of absorption and photoluminescence spectroscopy and dynamic light scattering. Photoinduced absorption spectroscopy was also employed to investigate interchain electronic communication and the photoinduced production of free charge carriers. The uncharged PPE had a relatively high polaron yield, indicating pi-stacking of adjacent PPE chains and efficient exciton splitting, while the charged polymer did not produce polarons, indicating that the polymers are not pi-stacked despite their tendency to form aggregates. This is most likely due to the presence of the cationic trimethylammonium side chains which force neighboring polymer chains too far apart to achieve effective pi-orbital overlap. Polarons were observed in both polymers after chemical doping with iodine. The ability to control aggregation and interchain electronic communication could be a useful tool in designing nanostructured electronic materials. PMID- 17078645 TI - Effective structure factor of osmotically deformed nanoemulsions. AB - Extreme osmotic compression of nanoemulsion droplets, achieved by ultracentrifugation, can create solidlike biliquid foams without causing significant droplet coalescence. Using small-angle neutron scattering (SANS), we probe the structure of a uniform silicone oil-in-water nanoemulsion stabilized by sodium dodecyl sulfate over a wide range of volume fractions, phi, up to and beyond the limit associated with maximal random jamming of spheres, phiMRJ = 0.64. Although some features in the structure can be understood at lower phi using simple predictions for hard spheres, the anionic repulsion and deformability of the droplet interfaces creates departures from these predictions at higher phi. For phi near and beyond phiMRJ, the effective structure factor, Seff, as a function of wavenumber, q, exhibits a primary peak that is subunity. We speculate that this striking feature is due to the deformation of the droplets into nonspherical shapes as the system begins to approach the limit of a random array of nanoscopic thin films, Plateau borders, and vertexes characteristic of a polyhedral foam. PMID- 17078647 TI - Models of adsorption at a line of three-phase contact. AB - Two model density distributions at a line of three-phase contact for which the adsorptions are readily calculated are analyzed. One of them provides a numerical illustration of a recently found surprising fact about the thermodynamics of adsorption at such contact lines. A form of the line analogue of the Gibbs adsorption equation is conjectured, and it is noted that the conjecture is in principle testable by computer simulation and by experiment. PMID- 17078646 TI - Shot-noise limited single-molecule FRET histograms: comparison between theory and experiments. AB - We describe a simple approach and present a straightforward numerical algorithm to compute the best fit shot-noise limited proximity ratio histogram (PRH) in single-molecule fluorescence resonant energy transfer diffusion experiments. The key ingredient is the use of the experimental burst size distribution, as obtained after burst search through the photon data streams. We show how the use of an alternated laser excitation scheme and a correspondingly optimized burst search algorithm eliminates several potential artifacts affecting the calculation of the best fit shot-noise limited PRH. This algorithm is tested extensively on simulations and simple experimental systems. We find that dsDNA data exhibit a wider PRH than expected from shot noise only and hypothetically account for it by assuming a small Gaussian distribution of distances with an average standard deviation of 1.6 A. Finally, we briefly mention the results of a future publication and illustrate them with a simple two-state model system (DNA hairpin), for which the kinetic transition rates between the open and closed conformations are extracted. PMID- 17078648 TI - Contact line dynamics in the late-stage coalescence of diethylene glycol drops. AB - We investigated the contact line dynamics of a composite drop formed as a result of the coalescence during the condensation of two diethylene glycol (DEG) drops at -4 degrees C on a silicon surface. The composite drop relaxes exponentially toward equilibrium with a typical relaxation time, tc, which depends on the equilibrium radius, R, of the composite drop. The value of tc is found to be in the range of 10-100 s for R approximately 1-4 microm. The relaxation dynamics is found to be larger by 6 orders of magnitude than that predicted by bulk hydrodynamics because of high dissipation in the contact line vicinity. Similar to low viscous liquids (water), this high dissipation can be attributed to an Arrhenius factor resulting from the phase change in the contact line vicinity and to the influence of surface defects that pin the contact line. PMID- 17078649 TI - Gemini surfactants at the air/water interface: a fully atomistic molecular dynamics study. AB - Gemini surfactants typically consist of two single-chain surfactants chemically linked by a spacer molecule. We report herein the results of fully atomistic molecular dynamics (MD) simulations of a series of Gemini surfactants: CsH2s alpha,omega-bis(C12H25N+(CH3)2Cl-), at the air/water interface with s = 3, 4, 6, 12, 14, and 16, at values of the initial surface area per surfactant AS = 70 A2, 77 A2, 95 A2, 151 A2, 133 A2, and 103 A2, respectively. The AS values employed were obtained from surface tension and neutron reflection experiments at the respective cmc of each surfactant. The Gemini surfactant corresponding to s = 3 was also simulated at AS = 105 A2, which is the experimentally derived value of surface area per surfactant at 1/10th of cmc. Only the surfactants with s = 12 and 14 and the surfactant with s = 3 at AS = 105 A2 gave a stable monolayer at the air/water interface. In other cases, we observe movement of some surfactant molecules from the air/water interface into the aqueous phase, resulting in a stable primary monolayer of surfactants at the air/water interface and a small concentration of surfactant molecules below it. The latter form aggregates, with their hydrophobic chains in the core. The density profiles along the normal to the interface are compared with the ones obtained from neutron reflection experiments. The MD simulations confirm the bending of the spacer toward the hydrophobic chains as the spacer length is increased and the spacer becomes more hydrophobic. The simulations have helped to shed light on the low-resolution picture which emerges from experimental analyses. PMID- 17078651 TI - Interaction of the neurotransmitter, neuropeptide Y, with phospholipid membranes: infrared spectroscopic characterization at the air/water interface. AB - The association of neuropeptide Y (NPY) at the air/water interface and with phospholipid monolayers on water as subphase has been investigated using external infrared reflection absorption spectroscopy (IRRAS). Studies of the conformation and orientation of NPY suggest that it adopts an alpha-helical structure and is oriented parallel to the air/water interface in neat peptide monolayers. Both secondary structure and orientation are preserved in mixed lipid/NPY monolayers. Comparison of NPY associated with zwitterionic DPPC and with anionic DMPS suggests that electrostatic attraction plays a major role for peptide binding to the membrane surface. PMID- 17078652 TI - Laser-induced local collapse in a langmuir monolayer. AB - Heating of a two-dimensional, methyloctadecanoate, Langmuir monolayer with a focused laser induces the local collapse of the monolayer. We observe the growth of a three-dimensional collapse aggregate that is fed by an inward flow of the two-dimensional monolayer surroundings. The experiments are explained with a hydrodynamic model describing the dynamics of the local collapse. From this theory we predict that local collapse can be induced if the collapse pressure of the monolayer decreases faster with temperature than with the surface tension of the pure air/water interface. Such conditions are fulfilled for lung surfactants, and it should therefore be possible to perform time-resolved local studies of the collapse of lung surfactants at those temperatures. PMID- 17078650 TI - Interaction of the neurotransmitter, neuropeptide Y, with phospholipid membranes: film balance and fluorescence microscopy studies. AB - The association of neuropeptide Y (NPY) with air-water interfaces and with phospholipid monolayers on water subphases and on physiological buffer has been investigated. Surface pressure (pi) versus molecular area (A) relations of the peptide at water surfaces depend on the concentration of the spreading solutions. Independent of that concentration, they show a transition from a low-density state to a high-density state at pi approximately 12 mN/m. Similar features are observed in the NPY adsorption to preformed monolayers (Deltapi(t --> infinity) as a function of pii = pi (t = 0) where t = 0 signifies the time of peptide injection). The transition is also observed in cospread lipid-NPY monolayers and is interpreted as the exclusion of the peptide from the surface layer. The reproducibility of the isotherms after expansion suggests that cospread lipid peptide monolayers are thermodynamically stable and that the peptide remains associated with the monolayer after exclusion from the lipid surface. A comparison of NPY association with zwitterionic and with anionic lipids as well as a comparison of the interactions on pure water and on physiological buffer suggest that electrostatic attraction plays a major role in the energetics of peptide binding to the membrane surface. Dual label fluorescence microscopy demonstrates that the peptide associates preferentially with the disordered, liquid condensed monolayer phase and also suggests that it self-aggregates upon exceeding a critical surface concentration. A NPY variant with a distorted alpha helix interacts with the surface as strongly as the natural NPY but expands the monolayers more. This suggests that the helix motif in the peptide is more important for the interaction with the receptor than for binding of the peptide to the membrane surface. In context, these observations attribute a specific role to the membrane in funneling the signal peptide to its membrane receptor. PMID- 17078653 TI - Raman spectroscopy of benzenethiolates on nanometer-scale gold clusters. AB - Near-infrared (1064-nm) irradiation of neat solid samples of benzenethiolate monolayer-protected gold clusters (MPCs) yields strong, well-resolved Raman spectra of the thiolate groups, comparable to those obtained for the same groups adsorbed at roughened gold electrodes. These clusters are formulated as TOAZ[AuN(SPh)M]Z-, N >> M and Z = 3-6, with core diameters of 1.7 and 1.5 nm, and were characterized previously by X-ray scattering, mass spectrometry, infrared spectroscopy, optical spectroscopy, nuclear magnetic resonance, and elemental analysis [Price, R. C.; Whetten, R. L. J. Am. Chem. Soc. 2005]. Numerous previous attempts to obtain spectra on various MPCs yielded only diffuse luminescence bands, as did benzenethiolate MPC samples of TOA2[Au44(SPh)28]2-, with 1.1-nm core diameters. The clusters are free of excess tetraoctylammonium bromide (TOABr) from the synthetic procedure, containing only the necessary TOA+ to maintain charge balance. In situ thermometry, using the anti-Stokes/Stokes intensity ratios, indicated the sample temperature remained below the onset of thermal decomposition. The Raman spectra of the clusters bear a strong resemblance to those obtained for nonmetallic (Au(I)SPh)x polymer samples that are not in resonant absorption at the laser wavelength. The smaller of the two cores, nominally TOA6[Au110(SPh)62], shows clearly a band at 505 cm(-1) assigned to a S-S stretch, suggestive of a moiety resembling diphenyl disulfide on the cluster surface. These results are interpreted with reference to recent reports suggesting a substantial "reconstruction" of the outermost gold layer upon thiolate adsorption (SAM formation). PMID- 17078654 TI - Conformational change of poly(ethylene glycol) near the critical point of isobutyric acid + water. AB - In solutions of isobutyric acid + water, poly(ethylene glycol) (PEG) can assume a helical conformation. [Alessi, M. L.; Norman, A. I.; Knowlton, S. E.; Ho, D. L.; Greer, S. C. Macromolecules 2005, 35, 9333-9340.] Here we report new measurements of the kinematic viscosity, nu, as a function of temperature for a solution of isobutyric acid + water at the critical composition, to which PEG (number average molecular weight = 1.01 x10(3) g/mol) was added at a concentration of 6.01 mg/mL. The data show that nu decreases near the critical point, with a maximum in nu at about 0.05 degrees C above the critical temperature, Tc. We interpret this change in nu in terms of a change in conformation of the polymer from helix to coil. This interpretation is supported by polarimetry measurements on the same mixture doped with (S)-(+)-1,2-propanediol, which indicates the loss of helicity near Tc and also a second helix-to-coil transition at about 60 degrees C. PMID- 17078655 TI - Grazing Incidence X-ray diffraction on Langmuir films: toward atomic resolution. AB - We have analyzed grazing incidence diffration (GIXD) data from condensed phases of Langmuir films of long-chain fatty acids at the air-water interface by using a new method consisting of a careful extraction of the structure factors followed by fitting of molecular parameters. We show that, contrary to the general belief, the information contained in GIXD spectra is enough to obtain near-atomic structural information. In particular, we directly determine for the first time the orientation of the chain backbone planes and of the carboxylic headgroups and we evaluate chain conformation defects. This new method allowed us to evidence a new phase of symmetry p2gm at high pressure, corresponding to a minimum in lattice energy, but never observed. PMID- 17078656 TI - A brief review of the relationships between monolayer viscosity, phase behavior, surface pressure, and temperature using a simple monolayer viscometer. AB - The two-dimensional surface shear viscosity, eta, of fatty acid monolayers of different chain lengths, measured using a simple magnetic needle viscometer, strongly correlates with the molecular organization in condensed phases and the absolute temperature. eta can increase by orders of magnitude at phase boundaries associated with tilted to untilted molecular order, providing the underlying order is semicrystalline. Hence, untilted, long-range ordered CS phases are the most viscous films. However, despite being untilted, the LS rotator phase is less viscous than certain laterally ordered tilted phases, suggesting a decrease of the van der Waals interactions due to molecular rotation. In certain regions of the L2 phase, eta reaches a maximum before the L2-LS transition, an anomalous behavior correlated with the change in the lattice symmetry of the headgroup. Surface shear viscosity, even when measured with a macroscopic probe, is particularly sensitive to the microscopic organization of monolayers. PMID- 17078657 TI - Subgel studies of dimyristoylphosphatidylcholine bilayers. AB - It is known that when bilayers of some saturated phosphatidylcholines are stored for 3 or more days at approximately 0 degrees C, a lamellar subgel (Lc) phase is detected at temperatures below the pretransition by differential scanning calorimetry (DSC). However, the subgel (Lc) phase and the corresponding subtransition (Lc--> Lbeta') for dimyristoylphosphatidylcholine (DMPC) has not been clearly characterized. In this study, using the temperature jump protocol first developed by Tristram-Nagle et al. for the dipalmitoylphosphatidylcholine (DPPC) system, new and accurate data characterizing the subgel formation and subtransition of DMPC were obtained through DSC and fluorescence spectroscopy with 1,6-diphenyl-1,3,5-hexatriene (DPH). It was discovered that the formation of the DMPC subgel phase requires incubation at temperatures of -5 degrees C or lower for 2 h or more. Kinetics of the subgel formation indicate that it is a very complex process and demonstrates that the planar gel phase is merely metastable below the subtransition, and not the thermodynamically stable phase. The subgel growth of DMPC is proven to be the dehydration of the headgroup region, and the subtransition is a process in which poorly hydrated DMPC becomes hydrated. PMID- 17078658 TI - Phase diagram for the adsorption of weak polyelectrolytes at a soft charged surface. AB - We have experimentally studied the adsorption of polyelectrolytes at oppositely charged surfaces. A weak flexible polyelectrolyte, poly(acrylic acid), was adsorbed from dilute solutions on a Langmuir film of a cationic amphiphile, dimethyldioctadecylammonium bromide. The polymer surface coverage, Gamma, at equilibrium was measured by two reflectivity techniques-ellipsometry and polarization modulated infrared reflection absorption spectroscopy (PM-IRRAS)-as a function of the surface charge density, sigma, and of the polymer ionization degree, alpha. Different adsorption regimes were evidenced. For weakly charged surfaces, sigma < sigma sat, Gamma increases with sigma and with 1/alpha, as expected for a neutralization of the surface by the adsorbed polymers. For highly charged surfaces, sigma > sigma sat, the adsorption of polyelectrolytes saturates. The mean orientation of the adsorbed chains also depends on the value of sigma: it is parallel to the surface for sigma < sigma (< sigma sat) and orthogonal to the surface for sigma > sigma. We have measured the values of sigma sat and sigma as a function of alpha and compared the results with existing theories. PMID- 17078659 TI - A low band gap conjugated polymer for supercapacitor devices. AB - We present the electrochemical polymerization and characterization of a stable and dopable polyindophenine derivative starting from monomer 5. The fabrication of supercapacitor devices was carried out by direct electrochemical polymerization on an ITO-coated slide. Furthermore, the galvanostatic and potentiostatic experiments conducted on these supercapacitors have shown that the polyindophenine-derivative material has very good cyclability, over a potential range of 1.4 V, and a maximum capacitance of 140 F g(-1). PMID- 17078660 TI - Tricritical phenomena in quasi-ternary mixtures of water + n-decane + n-undecane + tert-butanol. AB - Coexistence curves for the quasi-ternary system of water + n-decane + n-undecane + tert-butanol have been determined by measurements of the refractive index in three coexisting liquid phases. The binary mixtures of n-decane + n-undecane constructed the quasi-pure components in which the mass fraction beta of n-decane controls the approach to the tricritical point. The coexistence curves can be fitted to Scott's extended theory and can be extrapolated to a tricritical point at (44.1 +/- 0.3) degrees C and beta = 0.77 +/- 0.02 corresponding to an average n-alkane-carbon number of 10.19 +/- 0.02. The nonclassical critical amplitude ratio has been confirmed to be 4-5% smaller than the classical value, which is consistent with Fisher's prediction. PMID- 17078661 TI - Prediction of a highly activated state of CO adsorbed on an Al/Fe(100) bimetallic surface. AB - Using periodic slab density functional theory, we investigate CO adsorption, diffusion, and dissociation energetics on a monolayer of Al covering Fe(100) [Al/Fe(100)]. We predict a weakly chemisorbed state of CO to exist on Al/Fe(100), with CO adsorbing on the 4-fold hollow site in a very tilted fashion. This state is predicted to have an extremely low CO stretching frequency of only 883 cm(-1), indicating a dramatically weakened CO bond relative to gaseous CO, even though the molecule is predicted to bind to Al/Fe(100) quite weakly. We predict that dissociation of CO starting from this weakly adsorbed state has a barrier of only approximately 0.35 eV, which is approximately 0.70 eV lower than that on Fe(100). To understand how the underlying substrate changes the electronic properties of the supported Al monolayer, we compare CO adsorption on Al/Fe(100) to its adsorption on analogous pure Al(100) surfaces. This highly activated yet weakly bound state of CO on Al/Fe(100) suggests that Al/Fe(100) could be an effective low-temperature bimetallic catalyst in reducing environments. PMID- 17078662 TI - Interaction of the glycoalkaloid tomatine with DMPC and sterol monolayers studied by surface pressure measurements and Brewster angle microscopy. AB - The interaction of the glycoalkaloid tomatine with monolayers of dimyristoylphosphatidylcholine (DMPC) and cholesterol, as well as other selected sterols, has been investigated using surface pressure measurements at constant area and Brewster angle microscopy (BAM). The interaction of tomatine with sterol monolayers is found to vary with the structure of the sterol. The interaction of tomatine with cholesterol-containing monolayers results in a surface pressure increase accompanied by the appearance of a mottled texture. Morphological changes are observed that suggest the formation of tomatine-cholesterol complexes that aggregate at the water-air interface. No morphology change observable by BAM is observed for monolayers containing epicholesterol, suggesting that the stereochemistry of hydrogen bonding between the sterol and the sugar units on tomatine is of particular significance. Strong interactions are observed with cholestanol- and coprostanol-containing monolayers, and BAM reveals formation of spiked aggregates upon interaction with 7:3 mole ratio DMPC/coprostanol mixed monolayers. More modest surface pressure changes are observed for cholestenone- and epicoprostanol-containing monolayers. A much smaller surface pressure increase is observed when tomatine is injected beneath a pure DMPC monolayer. PMID- 17078663 TI - Predicting the phase diagram of two-dimensional colloidal systems with long-range interactions. AB - The phase diagram of a two-dimensional model system for colloidal particles at the air-water interface was determined using Monte Carlo computer simulations in the isothermic-isobaric ensemble. The micrometer-range binary colloidal interaction has been modeled by hard disklike particles interacting via a secondary minimum followed by a weaker longer-range repulsive maximum, both of the order of kBT. The repulsive part of the potential drives the clustering of particles at low densities and low temperatures. Pinned voids are formed at higher densities and intermediate values of the surface pressure. The analysis of isotherms, translational and orientational correlation functions as well as structure factor gives clear evidence of the presence of a melting first-order transition. However, the melting process can be also followed by a metastable route through a hexatic phase at low surface pressures and low temperatures, before crystalization occurs at higher surface pressure. PMID- 17078664 TI - Weak first-order tilting transition in monolayers of mono- and bipolar docosanol derivatives. AB - A systematic analysis of pressure-area isotherms and grazing incidence X-ray diffraction (GIXD) data of 22-methoxydocosan-1-ol (H3C-O-(CH2)22-OH, MDO), docosan-1-ol (H3C-(CH2)21-OH, DO), and docosyl methyl ether (H3C-(CH2)21-O-CH3, DME) monolayers on pure water between 10 and 35 degrees C is presented. All monolayers form fully condensed phases in the investigated temperature region. The GIXD data reveal that the monolayers exhibit the phase sequence -S at lower temperature and -LS at higher temperature. Phase diagrams have been established. Inserting a second hydrophilic group at the opposite end of the molecule (bipolar MDO) shifts the S/LS boundary to higher temperatures. All monolayers exhibit herringbone (HB) packing at lower temperatures. The "kink" in the isotherms observed at lower temperatures is replaced by a very small plateau region at higher temperatures. The entropy changes connected with this weak first-order tilting transition are much smaller compared with the first-order transition from liquid-expanded (LE) to condensed (LC). Additionally, this transition is endothermic in contrast to the LE/LC transition. The reason for the endothermic transition is the weaker positional correlation in the nontilted state compared with the tilted one. The appearance of the weak first-order endothermic transition can be connected with the changed phase sequence. X-ray photoelectron spectroscopy (XPS) measurements provide information about the polar group orientation. Considerations based on GIXD and XPS data as well as adhesion energy of the different terminal end groups lead to the conclusion that the hydroxyl group of the bipolar MDO is attached to the water surface while the methoxy group is in contact with air. The presented results show that the second hydrophilic group influences the monolayer properties in a mild way. PMID- 17078665 TI - Characterization of anomalous flow and phase behavior in a Langmuir monolayer of 2-hydroxy-tetracosanoic acid. AB - Langmuir monolayers of long-chain fatty acids display a wide range of interesting flow and phase behaviors, but the correlation between these behaviors varies. In this paper, we report two interesting behaviors in a Langmuir monolayer of 2 hydroxy-tetracosanoic acid: phase coexistence between the L(2h) and L(2)' phases over a large pressure range and a peak in the viscosity and elastic modulus of the material as a function of pressure. The two behaviors are confirmed over a range of temperatures for which the location of the undistorted lattice appears to vary relative to the range of coexistence for the two phases. Evidence for a correlation between the phase behavior and the peak in the mechanical properties is presented. PMID- 17078666 TI - Nucleation rates in a new phenomenological model. AB - In this paper we develop a new theory to evaluate the nucleation rate in the framework of the EMLD-DNT model. Beyond the model, our theory deals with cluster translation and exclusion, effects that have been virtually ignored in classical nucleation theory. We apply the model to the case of 1-pentanol, and compare the predictions with experimental results. We find an excellent agreement between the nucleation rate predicted by our theory and experimental data. The distinguishing feature of the model is its ability to predict successfully the rate of formation of the critical nucleus without the use of an intermolecular potential, employing only macroscopic thermodynamic properties. PMID- 17078667 TI - Effects of condensing agent and nuclease on the extent of ejection from phage lambda. AB - We have recently demonstrated, that DNA ejection from bacteriophage lambda can be partially or completely suppressed in vitro by external osmotic pressure. This suggests that DNA ejection from phage is driven by an internal mechanical force consisting of DNA bending and DNA-DNA electrostatic repulsion energies. In the present work we investigate the extent to which DNA ejection is incomplete at zero osmotic external pressure when phage is opened with its receptor in vitro. The DNA fragment remaining in the capsid and the tail that is no longer bent or compressed -and hence for which there is no internal driving force for ejection- is shown not to be ejected. We also demonstrate that DNA can be "pulled" out from the capsid by DNase I acting as a DNA binding protein or spermine acting as a DNA condensing agent. In particular, cryo electron microscopy and gel electrophoresis experiments show the following: (i) DNA ejection from bacteriophage lambda incubated in vitro with its receptor is incomplete at zero external osmotic force, with several persistence lengths of DNA remaining inside the phage capsid, if no nuclease (DNase I) or DNA condensing agent (spermine) is present in the host solution; (ii) in the presence of both DNase I and spermine in the host solution, 60% (approximately 29 kbp) of wild-type lambda DNA (48.5 kbp) remains unejected inside the phage capsid, in the form of an unconstrained toroidal condensate; (iii) with DNase I added, but no spermine, the ejection is complete; (iv) with spermine, but without DNase I added, all the DNA is again ejected, and organized as a toroidal condensate outside. PMID- 17078668 TI - Hydrogen sensors based on conductivity changes in polyaniline nanofibers. AB - Hydrogen causes a reversible decrease in the resistance of a thin film of camphorsulfonic acid doped polyaniline nanofibers. For a 1% mixture of hydrogen in nitrogen, a 3% decrease in resistance is observed (DeltaR/R = -3%). The hydrogen response is completely suppressed in the presence of humidity. In contrast, oxygen does not inhibit the hydrogen response. A deuterium isotope effect on the sensor response is observed in which hydrogen gives a larger response than deuterium: (DeltaR/R)H/(DeltaR/R)D = 4.1 +/- 0.4. Mass sensors using nanofiber films on a quartz crystal microbalance also showed a comparable deuterium isotope effect: DeltamH/DeltamD = 2.3 +/- 0.2 or DeltanH/DeltanD = 4.6 +/- 0.4 on a molar basis. The resistance change of polyaniline nanofibers is about an order of magnitude greater than conventional polyaniline, consistent with a porous, high-surface-area nanofibrillar film structure that allows for better gas diffusion into the film. A plausible mechanism involves hydrogen bonding to the amine nitrogens along the polyaniline backbone and subsequent dissociation. The inhibitory effect of humidity is consistent with a stronger interaction of water with the polyaniline active sites that bind to hydrogen. These data clearly demonstrate a significant interaction of hydrogen with doped polyaniline and may be relevant to recent claims of hydrogen storage by polyaniline. PMID- 17078669 TI - Triboelectrification between smooth metal surfaces coated with self-assembled monolayers (SAMs). AB - Using a modified surface forces apparatus, we have simultaneously measured the friction and triboelectrification between both similar and dissimilar molecularly smooth hexadecanethiol-coated metal surfaces on mica substrates. On shearing dissimilar surfaces, the tribocurrent increases dramatically as the load or pressure is increased, with large fluctuations about the mean. Neither charge transfer nor fluctuations are observed when the symmetric surfaces are sheared against each other. We also find that the type of friction, i.e., stick-slip or smooth sliding, the load and friction force, the sliding distance, and recent previous history have additional fine influences on the triboelectrification. Our results suggest that frictional dissipation induces electron-hole formation and charge transfer between two shearing surfaces due to molecular-level roughness and defects and local dielectric constant changes, giving rise to the observed tribocurrents. PMID- 17078670 TI - Polyamine-induced bundling of F-actin. AB - To better understand the mechanism of actin filament (F-actin) bundling by polyamines, we have measured the onset of bundling as a function of polyamine concentration. Samples were centrifuged at low speeds to separate bundles from unbundled actin, and the relative amounts of actin in the pellet and supernatant were determined via gel electrophoresis, yielding a description of the bundling transition as a function of actin and polyamine concentrations. These experiments were carried out for two different polyamines, spermine (tetravalent) and spermidine (trivalent). We found that the threshold concentration of polyamine needed to bundle actin is independent of both actin concentration and Mg2+ concentration over a wide range in Mg2+ concentration. We also find that spermine in F-actin bundles is essentially invisible in solution-phase proton NMR, suggesting that it is bound so tightly to F-actin that it is immobilized. PMID- 17078671 TI - Effects of temperature and chemical modification on polymer Langmuir films. AB - We previously reported on a rheological study of Langmuir films of poly(tert butyl methacrylate) (PtBMA), and this work describes further studies on this system. Here, surface pressure-area (Pi-A) isotherms and interfacial shear rheology experiments are performed to better understand the effects of two modifications: varying the temperature between 5 and 40 degrees C and introducing small amounts of carboxylic acid groups by partially hydrolyzing the polymer. Increased temperature produced isotherms with lower plateau surface pressures, Pip, and Pi values shifted to lower areas above Pip. Film properties transition from being primarily viscous to being dominated by elasticity as Pip is crossed for all temperatures studied, even as the value of Pip changes. Increasing the hydrolysis fraction leads to isotherms shifted to lower areas throughout the curve and higher Pip values. Both temperature variation and chemical modification are believed to affect the relative importance of polymer-polymer and polymer subphase interactions. PMID- 17078672 TI - Probing the limits of rate acceleration mediated by hydrogen bonds. AB - [Structure: see text] A simple receptor and substrate are used to probe the relationship between transition-state charge and the level of rate acceleration that can be created by stabilizing the transition state through hydrogen bonding. Pericyclic reactions are accelerated less than 2-fold by the receptor, whereas a conjugate addition reaction is accelerated more than 30-fold. Therefore, substrate polarization by hydrogen bonding would only appear to be effective for reactions that generate significant charge at the transition state. PMID- 17078673 TI - Functionalized oxepines via fragmentation of highly strained epoxides. AB - [Structure: see text] Epoxidation of highly strained cyclobutenes followed by thermal rearrangement provides a new entry into oxepine-containing bicyclo[5.3.0] ring systems. In contrast to the rearrangement of the corresponding cyclopropanated systems, the strained epoxides in this study are believed to fragment through two competing pathways leading to a mixture of diastereomeric 5 7 ring systems. PMID- 17078674 TI - Photoinduced electron transfer in a new Bis(C60)-phthalocyanine triad. AB - [Structure: see text] A novel covalently linked bis([60]fullerene)-phthalocyanine triad is reported, exhibiting long-lived photoinduced charge separation both in solution and in the solid state. The first demonstration of a working solar cell using triad 1 as the active material is also presented. PMID- 17078675 TI - Silylative Dieckmann-like cyclizations of ester-imides (and diesters). AB - [Structure: see text] Trialkylsilyl triflates effect cyclization of ester-imides such as 2 to produce adducts such as 4a. Trapping of the in situ generated, nucleophilic ketene acetal (cf. 5a) is a key aspect of the transformation. A range of substrates amenable to this operationally simple reaction is reported. In many instances the levels of diastereoselectivity are very high. Mechanistic points are inferred from spectroscopic observations. PMID- 17078676 TI - Benzo[a]acecorannulene: surprising formation of a new bowl-shaped aromatic hydrocarbon from an attempted synthesis of 1,2-diazadibenzo[d,m]corannulene. AB - [Structure: see text] Flash vacuum pyrolysis of 7,10-bis(2 bromophenyl)acenaphtho[1,2-d]pyridazine (C26H14Br2N2) has resulted in a surprising transformation, including dinitrogen loss, to give benzo[a]acecorannulene, a novel C26H12 bowl-shaped fullerene fragment. PMID- 17078677 TI - Palladium-catalyzed regiospecific aminocarbonylation of alkynes in the ionic liquid [bmim][Tf2N]. AB - [Structure: see text] Regiospecific construction of 2-substituted acrylamides was achieved by palladium-catalyzed aminocarbonylation of alkynes in the ionic liquid [bmim][Tf2N] without any acid additive under relatively mild conditions. The ionic liquid was used as the reaction medium and also acted as a promoter. Acrylamides were obtained in moderate to excellent yields, and an important feature is that the catalyst system can be recycled five times without loss of catalytic activity. PMID- 17078678 TI - Synthesis of plakortone B and analogs. AB - [Structure: see text] Use of a palladium-mediated alkoxycarbonylation/lactonization process provides a variable route to analogs of the plakortones. Four different analogs, including natural plakortone B, have been synthesized via this route. PMID- 17078679 TI - Kalanchosides A-C, new cytotoxic bufadienolides from the aerial parts of Kalanchoe gracilis. AB - [Structure: see text] Three new compounds, kalanchosides A-C (1-3), as well as five known compounds, were isolated from the aerial parts of Kalanchoe gracilis. The compound structures were determined by spectroscopic methods. All eight isolated compounds showed significant cytotoxic activity against a panel of human tumor cell lines, with potency reaching the nanomolar range. However, only bryophyllin B (8) inhibited HIV replication in H9 lymphocyte cells. PMID- 17078680 TI - Direct palladium-catalyzed ortho-arylation of benzylamines. AB - [Structure: see text] Unsubstituted benzylamines and N-methylbenzylamine can be ortho-arylated under palladium catalysis at 130 degrees C. The reactions require the presence of trifluoroacetic acid and silver acetate. PMID- 17078681 TI - Powerful Ti-crossed Claisen condensation between ketene silyl acetals or thioacetals and acid chlorides or acids. AB - [Structure: see text] A powerful Ti-crossed Claisen condensation between ketene silyl acetals (KSAs) and acid chlorides was successfully performed to give alpha monoalkylated esters and thermodynamically unfavorable (less accessible) alpha,alpha-dialkylated beta-keto esters in good yield (46 examples; 41-98% yield). A closely related reaction between ketene silyl thioacetals (KSTAs) and acid chlorides also proceeded smoothly to give alpha-monoalkylated and alpha,alpha-dialkylated beta-keto thioesters (21 examples; 61-97% yield). The present protocol was extended to the direct condensation of KSAs with carboxylic acids (14 examples; 71-97% yield). PMID- 17078682 TI - Synthesis of the entire framework of tartrolon B utilizing a silicon-tethered ring-closing metathesis strategy. AB - [Structure: see text] A tandem ring-closing metathesis (RCM) of silaketal tethered dienynes gives rise to bicyclic siloxanes, which upon removal of the silicon tether afford dienediol skeletons with a stereodefined E,Z-1,3-diene motif. The implementation of this methodology has led to the construction of the entire C1-C21 linear carbon skeleton of tartrolon B. PMID- 17078683 TI - Stereoselective synthesis of the phorboxazole A macrolide by ring-closing metathesis. AB - [Structure: see text] Described is a regio- and stereoselective ring-closing metathesis (RCM) to form the C2-C3 alkene of the macrolide-containing domain of phorboxazole A. This work demonstrates a dramatic effect of reaction solvent on RCM product (E/Z)-selectivity. This process offers an alternative assembly of the macrolide-containing domain of phorboxazole A, one of the most potent anticancer agents known. PMID- 17078684 TI - Luminescence tuning of organoboron quinolates through substituent variation at the 5-position of the quinolato moiety. AB - [Structure: see text] A series of organoboron quinolates with emission colors ranging from blue to red have been prepared. In comparison to the respective AlQ3 derivatives a distinct blue-shift of the emission is observed. Theoretical calculations serve to provide insight into the nature of the frontier orbitals and the effect of the substituents in the 5-position of the quinolate ligands on the relative HOMO and LUMO energy levels. An efficient new blue emitting material with a pinacolborane substituent has been identified. PMID- 17078685 TI - Fully automated continuous flow synthesis of 4,5-disubstituted oxazoles. AB - [Structure: see text] A multipurpose mesofluidic flow reactor capable of producing gram quantities of material has been developed as an automated synthesis platform for the rapid on-demand synthesis of key building blocks and small exploratory libraries. The reactor is configured to provide the maximum flexibility for screening of reaction parameters that incorporate on-chip mixing and columns of solid supported reagents to expedite the chemical syntheses. PMID- 17078686 TI - Preparation, properties, and reduction of a novel TCNQ-type thienoquinoid. AB - [Structure: see text] TCNQ-type bithienoquinoid condensed with a alpha-dicarbonyl group was synthesized. The compound revealed a reversible reduction wave at +0.32 V vs SCE. The radical anion species was cleanly generated by a reduction with Na Hg and characterized: lambdamax=1027 nm in vis-NIR and a large spin density of the beta-carbon atoms on the fused ring by EPR spectroscopy. The radical anion salts were isolated by a reduction with iodide and showed moderate conductivity. PMID- 17078687 TI - Substrate-directed stereoselectivity in vicinal diamine-catalyzed synthesis of warfarin. AB - [Structure: see text] A new mechanism involving a diimine intermediate is proposed for vicinal diamine-catalyzed synthesis of warfarin. Decreasing the NCCN dihedral angle by varying the diamine results in an increase in the enantioselectivity of warfarin synthesis. PMID- 17078688 TI - From oligomers to polymer: convergence in the HOMO-LUMO gaps of conjugated oligomers. AB - [Structure: see text] Extrapolation of HOMO-LUMO gaps for pi-conjugated oligomers at the B3LYP/6-31G(d) level of theory predict accurately (within 0.1-0.2 eV) the band gaps of conjugated polymers only when long (at least 20-mer) pi-conjugated oligomers are used for the extrapolation. PMID- 17078689 TI - Synthesis, characterization, and utility of thermoresponsive natural/unnatural product macroligands for affinity chromatography. AB - [Structure: see text] The synthesis and characterization of thermoresponsive, water-soluble poly-N-isopropyl acrylamide (PNIPAM) derived macroligands displaying cyclosporin A (CsA) and dexamethasone (Dex) for use as novel affinity resins are described. Characterization of these soluble macroligands, including ligand loading and integrity, was determined by 1H NMR spectroscopy. One of the CsA macroligands was used in a protein affinity experiment to capture known binding proteins of CsA, the cyclophilins, from Jurkat T-cell lysates. PMID- 17078690 TI - Synthesis of primary arylselenoamides by reaction of aryl nitriles with Woollins' reagent. AB - [Structure: see text] The reaction of aryl nitriles with Woollins' reagent followed by water affords a variety of primary arylselenoamides in 60-100% yield. The first crystal structures of two primary selenoamides are reported. PMID- 17078691 TI - Synthesis and incorporation of a cationic pyridinium nucleoside mimic in duplex DNA: effect on long-distance radical cation transport. AB - [Structure: see text] A convenient method for the synthesis and solid-phase incorporation of a cationic nucleoside mimic and its role in DNA long-range charge transfer are described. PMID- 17078692 TI - Structural impact of coordinating a helical 2,6-pyridyl diamide with divalent metals. AB - [Structure: see text] The structural consequences of coordinating 2,6-bis[2-((4S) 4-methyl-4,5-dihydro-1,3-oxazol-2-yl)phenyl]carbamoylpyridines, 2, with divalent metals such as Cu(II), Ni(II), and Zn(II) are reported. Metal coordination occurs under mild conditions in a manner that preserves the helical bias of the parent ligand in the solid state and in solution. 1H NMR line-shape analysis indicates that metal coordination increases the helical interconversion barrier, thus rigidifying the dynamic helicity of 2. PMID- 17078693 TI - Tetraphenyloxybenziporphyrin, a new organometallic ligand for silver(III) and gold(III). AB - [Structure: see text] Reaction of 4-hydroxyisophthalaldehyde with excess phenyl magnesium bromide gave a dicarbinol and this condensed with pyrrole and aromatic aldehydes in the presence of BF3.Et2O to afford, following oxidation with DDQ, novel tetraarylcarbaporphyrinoids in 10-24% yield. Further reaction with silver(I) acetate or gold(III) acetate gave stable organometallic derivatives that retained the aromatic characteristics of the parent macrocycle. PMID- 17078694 TI - Isokibdelones: novel heterocyclic polyketides from a Kibdelosporangium sp. AB - [Structure: see text] The isokibdelones are an unprecedented family of polyketides produced by an Australian isolate of a rare actinomycete, Kibdelosporangium sp. The structures of the isokibdelones were assigned by spectroscopic analysis and chemical interconversion. A proposed biosynthesis requires a novel molecular twist that generates an unprecedented heterocyclic system and differentiates the isokibdelones from their kibdelone co-metabolites. SAR analysis on the isokibdelones further defines the anticancer pharmacophore of these novel polyketides. PMID- 17078695 TI - Tandem semipinacol/Schmidt reaction leading to a versatile and efficient approach to azaquaternary alkaloid skeletons. AB - [Structure: see text] A TiCl4-promoted tandem semipinacol/Aube's type intramolecular Schmidt reaction of alpha-siloxy-epoxy-azide has been designed and developed to be a general method for efficient construction of azaquaternary carbon units. As applicable examples, some key tricyclic azaquaternary skeletons incorporated in many important alkaloids, such as cephalotaxine, stemonamine, erythrinan, and homoerythrinan alkaloids, have been constructed. PMID- 17078696 TI - Straightforward and highly efficient catalyst-free one-pot synthesis of dithiocarbamates under solvent-free conditions. AB - [Structure: see text] A highly efficient and simple synthesis of dithiocarbamates is possible based on the one-pot reaction of amines, CS2, and alkyl halides without using a catalyst under solvent-free conditions. The mild reaction conditions, high yields, and broad scope of the reaction illustrate the good synthetic utility of this method. The reaction is a highly atom-economic process for production of S-alkyl thiocarbamates and successfully can be used in high quantities in the pharmaceutical or agrochemical industries. PMID- 17078697 TI - Stereoselective total synthesis of the proposed structure of 2-epibotcinolide. AB - [Structure: see text] The total synthesis of pseudo 2-epibotcinolide (1b) through several featured synthetic approaches has been attained. First, the chiral linear precursors of the nine-membered ring compound is stereoselectively constructed by the asymmetric aldol reaction for producing beta-hydroxy ester units. Second, the key cyclization reaction to form the nine-membered lactone moiety is efficiently achieved by the extremely facile and powerful mixed-anhydride method promoted by 2-methyl-6-nitrobenzoic anhydride (MNBA) with basic promoters. PMID- 17078698 TI - Synthesis of (+/-)-clusianone: high-yielding bridgehead and diketone substitutions by regioselective lithiation of enol ether derivatives of bicyclo[3.3.1]nonane-2,4,9-triones. AB - [Structure: see text] A concise synthesis of the polyprenylated acylphloroglucinol natural product, clusianone, in racemic form, is described. An Effenburger cyclization generated a core bicyclo[3.3.1]nonane-trione structure, which was then elaborated by means of regioselective lithiation reactions. PMID- 17078699 TI - A synthesis of the welwistatin core. AB - [Structure: see text] An approach to the core structure of the microtubule targeting agent welwistatin is described. The approach utilizes the type 2 intramolecular Diels-Alder reaction, with indole serving as the tether between diene and dienophile, to form the natural product's characteristic bicyclo[4.3.1]decane skeleton. PMID- 17078700 TI - Manganese(III) acetate promoted regioselective phosphonation of heteroaryl compounds. AB - [Structure: see text] A new method for direct phosphonation of thiazoles, furans, and pyrroles is introduced. Reactions of the heteroaryl compounds with dimethyl or diethyl phosphites and Mn(OAc)(3).2H2O under mild conditions give phosphonated products in high yield and good regioselectivity. PMID- 17078701 TI - Site-specific preparation of 2-carboalkoxy-4-substituted naphthalenes and 9 alkylphenanthrenes and evidence for an allene intermediate in the novel base catalyzed cyclization of 2-alkynylbiphenyls. AB - [Structure: see text] A site-specific preparation of 2-carboalkoxy-4-substituted naphthalenes and 9-alkylphenanthrenes is described. The successful cyclization of an allene intermediate provides supportive evidence for the previously proposed mechanism. PMID- 17078702 TI - Total synthesis and initial biological evaluation of new B-ring-modified bryostatin analogs. AB - [Structure: see text] The total synthesis and preliminary biological evaluation of the first bryostatin analogs (bryologs) to incorporate B-ring substitution are reported. Asymmetric syntheses of two new polyketide "spacer" domains are described, one exploiting the pseudosymmetry of the C1-C13 region. These fragments are convergently joined to the "recognition" domain through a remarkably versatile macrotransacetalization process. The resulting new analogs exhibit potent nanomolar or picomolar affinity to protein kinase C (PKC), comparable to or better than that found for bryostatin. PMID- 17078703 TI - Room temperature hydroamination of N-alkenyl ureas catalyzed by a gold(i) N heterocyclic carbene complex. AB - [Structure: see text] Treatment of an N-4-pentenyl or N-5-hexenyl urea with a catalytic 1:1 mixture of a gold(I) N,N-diaryl imidazol-2-ylidine complex and AgOTf at or near room temperature leads to intramolecular exo-hydroamination to form the corresponding nitrogen heterocycle in excellent yield. PMID- 17078704 TI - Total synthesis and stereochemistry of the antitumor antibiotic PD 113,271. AB - [Structure: see text] A total synthesis of PD 113,271, an antitumor fostriecin analogue isolated from Streptomyces pulveraceus, was achieved by the chiral pool approach starting with D-galactose and L-tartaric acid. The synthesis of PD 113,271 led to unambiguous assignment of the relative and absolute stereochemistry of its stereocenters. PMID- 17078705 TI - Total synthesis of (+/-)-morphine. AB - [Structure: see text] The morphinan skeleton was effectively synthesized by an intramolecular Mannich-type reaction. Further transformation led to total synthesis of morphine. PMID- 17078706 TI - Use of allylic strain to enforce stereochemistry. Direct syntheses of 7,8 dihydroxycalamenene and mansonone C. AB - [Structure: see text] Direct syntheses of 7,8-dihydroxycalamenene and mansonone C were achieved. The cis-stereochemistry required for the synthesis of 7,8 dihydroxycalamenene was introduced by an intramolecular cyclization directed by allylic strain. PMID- 17078707 TI - Lewis acid catalyzed reaction of arylvinylidenecyclopropanes with acetals: a facile synthetic protocol for the preparation of indene derivatives. AB - [Structure: see text] A number of highly substituted indene derivatives have been prepared in good yields by the reactions of arylvinylidenecyclopropanes 1 with acetals 2 in the presence of Lewis acid under mild conditions. The reaction is believed to proceed via regioselective addition of oxonium intermediate to arylvinylidenecyclopropane and the subsequent intramolecular Friedel-Crafts reaction. PMID- 17078708 TI - Qualitative and quantitative measurements of hydrogen bond mediated scalar couplings in acyclic 1,3-diols. AB - [Structure: see text] OH...OH hydrogen bond mediated scalar couplings have been observed in acyclic syn- and anti-1,3-diols using a 2D 1H COSYLR NMR experiment and quantified with an uncertainty of +/-0.02 Hz with a selective-excitation spin echo NMR experiment. A theoretical investigation confirmed the importance of the hydrogen bond in mediating the spin-spin coupling in these systems. PMID- 17078709 TI - Azabicyclic amino acids by stereoselective dearomatizing cyclization of the enolates of N-nicotinoyl glycine derivatives. AB - [Structure: see text] On activation by pyridine N-acylation, enolates of N nicotinoyl and N-isonicotinoyl glycine and alanine derivatives cyclize to yield 6,5-azabicyclic or 6,4-azaspirocyclic lactams. With an N-alpha-methyl-p methoxybenzyl group the cyclization is diastereoselective; hydrogenation and deprotection yields azabicyclic amino acids in 94:6 er. PMID- 17078710 TI - Biosynthesis of streptolydigin: origin of the oxygen atoms. AB - [Structure: see text] Biosynthetic incorporations of [13C,18O]-labeled precursors into streptolydigin reveal the origin of the oxygen atoms, limiting the possible pathways for acyltetramic acid ring assembly and for formation of the bicyclic ketal. PMID- 17078711 TI - High turnover frequency observed in catalytic enantioselective additions of enecarbamates and enamides to iminophosphonates. AB - [Structure: see text] In addition reactions of enecarbamates and enamides, extremely high turnover frequency of the catalyst was observed in comparison with that of silicon enolate addition reactions. This is presumably due to fast transfer of the proton that locates on the nucleophiles. PMID- 17078712 TI - Revision of the structures of citrifolinin A, citrifolinoside, yopaaoside A, yopaaoside B, and morindacin, iridoids from Morinda citrifolia L. and Morinda coreia Ham. AB - [Structure: see text] Citrifolin A was revised to dehydromethoxygaertneroside (2). Citrifolinoside and yopaaoside A were found to be identical, and their structures were revised to the new structure dehydroepoxymethoxygaertneroside (5). Yopaaoside B was revised to citrifolinoside A (9), and morindacin was revised to borreriagenin (11). PMID- 17078713 TI - Exploitation of cyclopropane ring-cleavage reactions for the rapid assembly of tetracyclic frameworks related to gibberellins. AB - [Structure: see text] The readily available hexahydrofluorene 5 has been elaborated over six steps, including three involving cyclopropane ring-cleavage reactions, into compound 12 which incorporates the carbocyclic framework associated with gibberellins. PMID- 17078714 TI - Glycerol nucleoside triphosphates: synthesis and polymerase substrate activities. AB - [Structure: see text] The synthesis of (S)-glycerol nucleoside triphosphates (gNTPs) and the analysis of their substrate activities for enzymatic polymerization is described. NTPs with simplified carbohydrate backbones such as the tNTPs (alpha-L-threose-NTPs) are polymerase substrates and offer the potential to create non-natural aptamer sequences with simplified backbones through enzymatic means. The acyclic (S)-GNA was modeled after the shortened alpha-threofuranosyl backbone. Here we describe the synthesis of (S)-glycerol NTPs and initial enzymatic testing of this further simplified nucleic acid backbone. PMID- 17078715 TI - Cyclization reactions of homopropargyl azide derivatives catalyzed by PtCl4 in ethanol solution: synthesis of functionalized pyrrole derivatives. AB - [Structure: see text] PtCl4-catalyzed cyclization reactions of homopropargyl azide derivatives to pyrrole rings were investigated. Using ethanol as solvent with 2,6-di-tert-butyl-4-methylpyridine as the base was found to be the best set of conditions for effecting this ring-closing reaction. These reaction conditions can be applied to the preparation of functionalized pyrrole derivatives, with no effect on the functional groups. PMID- 17078716 TI - Enantioselective equilibration-access to chiral aldol adducts of mandelic acid esters. AB - [Structure: see text] Syn-configured aldol products of mandelic acid esters and aldehydes were synthesized by the catalytic use of amines in the presence of titanium(IV) tert-butoxide. Used along with chiral N-methylephedrine, anti configured alpha,beta-dihydroxyesters were isolated with a high degree of enantioselectivity for the first time. PMID- 17078717 TI - Beta-isocupreidine-catalyzed Baylis-Hillman reaction of chiral N-boc-alpha-amino aldehydes. AB - [Structure: see text] Beta-isocupreidine (beta-ICD)-catalyzed Baylis-Hillman reaction of chiral N-Boc-alpha-amino aldehydes and 1,1,1,3,3,3 hexafluoroisopropyl acrylate (HFIPA) takes place without racemization and exhibits the match-mismatch relationship between the substrate and the catalyst. In the case of acyclic amino aldehydes, L-substrates show excellent syn selectivity and high reactivity in contrast to D-substrates. On the other hand, in the case of cyclic amino aldehydes, D-substrates rather than L-substrates show excellent anti selectivity and high reactivity. PMID- 17078718 TI - AuCl-catalyzed [4+2] benzannulation between o-alkynyl(oxo)benzene and benzyne. AB - [Structure: see text] The AuCl-catalyzed benzannulation of o-alkynyl(oxo)benzenes with benzenediazonium 2-carboxylate proceeds under mild conditions and a variety of anthracene derivatives, having a ketone group at the 9-position, are produced in good to high yields. The reaction proceeds most probably through the [4+2] cycloaddition between benzyne and benzopyrylium auric ate complex, which would be generated by the gold-induced electrophilic cyclization of o alkynyl(oxo)benzenes. PMID- 17078719 TI - General and highly efficient synthesis of 2-alkylideneazetidines and beta-lactams via copper-catalyzed intramolecular N-vinylation. AB - [Structure: see text] N-Tosyl-3-halo-3-butenylamines underwent efficient Ullmann type coupling with the catalysis of CuI/N,N'-dimethylethylenediamine to afford 2 alkylideneazetidines, which could be readily converted to the corresponding beta lactams by oxidation with O3. PMID- 17078720 TI - Diastereoselective multicomponent synthesis of dihydropyridones with an isocyanide functionality. AB - [Structure: see text] In a search for new multicomponent strategies leading to valuable small heterocycles, a new highly diastereoselective four-component reaction (4CR) was found in which a phosphonate, nitriles, aldehydes, and isocyanoacetates combine to afford functionalized 3-isocyano-3,4-dihydro-2 pyridones. In this strategy, initially a 1-azadiene is generated, which is trapped in the same pot by an isocyanoacetate as the fourth component. Multicomponent reactions (MCRs) that lead to heterocycles containing isocyano substituents are unprecedented and offer many possibilities for further differentiation. PMID- 17078721 TI - Studies on oxidopyrylium [5+2] cycloadditions: toward a general synthetic route to the C12-hydroxydaphnetoxins. AB - [Structure: see text] 12-hydroxydaphnetoxins, members of the structurally fascinating daphnane diterpene family, exhibit a wide range of significant biological activities. A general route to the BC-ring system of 12-hydroxy daphnetoxins is reported based on D-ribose. Depending on the choice of protecting groups and solvent, the oxidopyrylium-alkene [5+2] cycloaddition originating from A provides cycloadduct diastereomer B or C with good to excellent selectivity. PMID- 17078722 TI - Switchable dual binding mode molecular shuttle. AB - [Structure: see text] Protonation controls the location of a dual binding mode macrocycle in a [2]rotaxane. In the neutral form, amide-amide hydrogen bonds hold the macrocycle over a dipeptide residue; when the thread is protonated, polyether ammonium cation interactions dominate and the macrocycle changes position. PMID- 17078724 TI - Efficient preparation of proline N-carboxyanhydride using polymer-supported bases. AB - [Structure: see text] A procedure for the preparation of proline N carboxyanhydride in high yield and purity is described using polymer-supported tertiary amines. The polymer-supported amine can be recycled with a basic wash and filtration of the resin. The procedure facilitates the access to the efficient preparation of the polyproline polymer with potential therapeutic interest. PMID- 17078723 TI - Electronegative oligothiophenes based on a hexafluorocyclopentene-annelated thiophene unit. AB - [Structure: see text] The synthesis of hexafluorocyclopenta[c]thiophene and its based oligothiophenes is described. The effectiveness of a hexafluorocyclopentene unit to lower the LUMO level without disturbing the effective conjugation could be unambiguously clarified by spectroscopic measurements and X-ray analysis. PMID- 17078725 TI - Direct C-H arylation of purines: development of methodology and its use in regioselective synthesis of 2,6,8-trisubstituted purines. AB - [Structure: see text] Direct C-H arylation of purines to position 8 by diverse aryl iodides was achieved with Pd catalysis in the presence of CuI and Cs2CO3. The methodology is general and efficient and was applied in the consecutive regioselective synthesis of 2,6,8-trisubstituted purines bearing three different C-substituents in combination with two cross-coupling reactions. PMID- 17078727 TI - It is "thiazolidene-2-imine" and not imidazole-2-thione as the reaction product of 1-benzoyl-3-phenylthiourea with Br(2)/enolizable ketone. AB - [Structure: see text] The products obtained by the reaction of benzoyl-3 phenylthioureas with bromine and enolizable ketones in the presence of triethylamine are not imidazole-2-thione derivatives as reported (Org. Lett. 2003, 5, 1657-1659) rather they are thiazolidene-2-imine derivatives. PMID- 17078726 TI - Electrophile-induced ether transfer: a new approach to polyketide structural units. AB - [Structure: see text] A strategically novel approach to the formation of syn-1,3 diol mono- and diethers through electrophilic activation of homoallylic alkoxymethyl ethers has been developed. The resulting polyketide-like synthetic fragments are generated in good yield and with excellent stereocontrol. A chairlike transition state is proposed to account for the high stereoselectivity. Varying the conditions of the reaction workup results in the efficient generation of mono- and diether containing structural units common to polyketide natural products. PMID- 17078728 TI - Total synthesis of the chlorinated marine natural product dysamide B. AB - [Structure: see text] Two approaches to the synthesis of (2S,4S)-5,5 dichloroleucine are compared, and the parent amino acid was used in the first total synthesis of the polychlorinated marine natural product, dysamide B. A key step was the lead tetraacetate-mediated decarboxylation of an alpha,alpha dichloro acid in the presence of 1,4-cyclohexadiene to generate the dichloromethyl group. PMID- 17078729 TI - Copper-catalyzed asymmetric conjugate addition of diethylzinc to alpha,beta unsaturated imines derived from alpha-aminoacids. Enantioselective synthesis of gamma-substituted alpha-dehydroaminoesters. AB - [Structure: see text] A highly enantioselective synthesis of alpha dehydroaminoacids with a stereogenic center in the gamma position through copper catalyzed asymmetric conjugate addition of diethylzinc to alpha,beta-unsaturated imines using a TADDOL-derived phosphoramidite complex is reported. PMID- 17078730 TI - Palladium-catalyzed preparation of vinylallenes from 2-bromo-1,3,5-trienes via an alkylidene-pi-allylpalladium-mediated formal SN2" pathway. AB - [Structure: see text] A novel Pd-catalyzed reaction to prepare conjugated vinylallenes from 2-bromo-1,3,5-triene and a soft nucleophile via a formal SN2" pathway was developed. The reaction proceeds via alkylidene-pi-allylpalladium and allenyl-pi-allylpalladium intermediates, and a dynamic process involving the two palladium intermediates played important roles in determining the selectivity of the Pd-catalyzed reaction. The reaction was extended to an asymmetric counterpart, and an axially chiral vinylallene was obtained with up to 81% ee. PMID- 17078731 TI - Reactivity and selectivity of 1,3-diyn-6-enes in electrophilic transition metal catalyzed reactions. AB - [Structure: see text] 1,3-diyne is an excellent source of alkynyl metal carbene species upon activation with an electrophilic metal catalyst. The products from this bond reorganization process suggest that the metal carbene species, generated from the preferential participation of an acetate over an alkene in the first step, undergo an efficient metallotropic [1,3]-shift followed by termination via cyclopropanation. PMID- 17078732 TI - Enantioselective organocatalytic aldol reaction of ynones and its synthetic applications. AB - [Structure: see text] For the first time, unmodified ynones were used in organocatalytic asymmetric aldol reactions delivering monoprotected anti alpha,beta-dihydroxyynones in high yields, dr's up to 19:1, and ee's up to 95%. These products can be either reduced to afford enantioenriched unsaturated anti,anti-triol or cyclized using a novel intramolecular phosphine-catalyzed alpha-addition to the ynone. This organocatalytic sequential aldol-cyclization process provides a concise entry to unusual enantioenriched oxygenated heterocycles, which can be used for subsequent structural manipulations. PMID- 17078733 TI - Multiple agenesis and anhidrotic ectodermal dysplasia: a comparative longitudinal study of dental similarities and genetic differences in two groups of children. AB - AIM: Dental anomalies in shape and number may be present isolated or associated with other manifestations. In anhidrotic ectodermal dysplasia they occur more frequently and severely. The authors examined a group of children with similar dental anomalies but no other ectodermal or extra-ectodermal signs. METHODS: This study makes a comparative evaluation of similarities and differences of dental anomalies between two groups: A anhidrotic ectodermal dysplasia and B similar dental finding but without extra- dental anomalies. RESULTS: In group A, the average number of agenesis in primary teeth was 3.5 (upper) and 5.33 (lower); in permanent teeth it was 5.4 and 5.8, respectively. In group B, the average was 1.62 (upper) and 0.25 (lower) in primary teeth, and 4.0 and 4.25 in permanent teeth respectively, with no constant pattern of occurrence. The study of tooth morphology of both groups revealed numerous anomalies in both dentitions. No differences were found in the average number of agenesis and morphological anomalies in the permanent teeth between both groups, but in the primary dentition group B presented a lower degree of incidence. CONCLUSION: The presence of almost normal primary dentition (regarding to number), but with morphological anomalies, should lead to suspect their exacerbation in the permanent dentition. PMID- 17078734 TI - Clinical evidence and literature to compare two different therapeutic protocols in tooth avulsion. AB - AIM: This is to assess the priority in tooth avulsion: replantation as quickly as possible and deferred endodontic treatment, or replantation and elimination of every irritating stimulus for the periodontal ligament. The objective is to establish a univocal protocol by analysing and comparing the various components of these therapeutic procedures. MATERIALS AND METHODS: STUDY DESIGN: 20 patients were selected and divided into 2 groups: in group A we focused our attention on the rapidity of replantation, in group B we focused on the elimination of the necrotic pulp and every possible irritating stimulus that might lead to root resorption of inflammatory nature. RESULTS: Clinically, dental mobility seems to be greater in the first group. However after 15 days teeth have a comparable mobility and after one month they all show a mobility degree 1. Radiographically a greater incidence of resorption can be observed in group A compared to group B as in the first one a higher number of replanted teeth undergo resorption. Such lesions have the tendency to remain constant or even increase. CONCLUSIONS: The results of this study seem to take only one direction: extraoral endodontic therapy. PMID- 17078735 TI - Cementoenamel junction of deciduous teeth: SEM-morphology. AB - AIM: The aim of this study was to investigate the cementoenamel junction of a group of 11 primary sound mandibular incisors extracted for orthodontic reasons. MATERIALS AND METHODS: Eleven caries and defect-free human inferior deciduous incisors were extracted for orthodontic reasons and the cementoenamel junction was investigated by scanning electron microscopy (SEM). The types of tissue interrelations were classified in four possible categories: 1) cementum and enamel edge-to-edge, 2) cementum overlapped by enamel, 3) enamel overlapped by cementum, 4) presence of exposed dentin between enamel and cementum. RESULTS: In our observations root cementum and enamel edge-to-edge interrelation was the most frequent feature observed in overall sample, root cementum overlapping enamel tissues was observed in more than one third of the cementoenamel junction area, exposed dentin was a rare observation. In few, small and rare areas enamel overlapped cementum. Further studies could determine statistical prevalence. CONCLUSION: The cementoenamel junction of primary teeth differs from that of permanent teeth, the scarcity of gaps between cementum and enamel, the epithelial junction at the equator of the crown and the globosity of the crown are probable protective factors toward decay susceptibility. PMID- 17078736 TI - Oral hygiene habits and attitudes in a school population in L'Aquila (Italy). AB - AIM: This study aims to analyse the results of a questionnaire submitted to students of the elementary schools in L'Aquila in order to understand their awareness of a correct caries prevention method by using fluoride. MATERIALS AND METHODS: 4th and 5th grade students of elementary schools in L'Aquila were submitted to a questionnaire; from a total of 1205 students, age ranging from 8 to 12, 1094 were questioned; 7 children 8 years old, 282 of 9, 555 of 10, 243 of 11, 8 of 12, and only 2 of 13, respectively (the last 2 were registered later at school because non-EU citizens). RESULTS: Among the Italian population with a low incidence of dental decay, a relatively limited group, called 'high risk decay group', revealed a higher incidence of the pathology than the standard one. CONCLUSION: In a multiracial society, as ours, it is important to carry out an effective screening in order to recognize different categories of patients through simple questions inserted in the patient's case history. PMID- 17078737 TI - The association between bronchial asthma and dental caries in children of the developmental age. AB - AIM: The aim of the study was to evaluate the dental caries experience of children suffering from bronchial asthma. METHODS: During the year 1998 a clinical dental examination was conducted in the northern part of Poland on children aged 3-15 years who had chronic bronchial asthma. The control group comprised 326 generally healthy children matched for age, sex and socioeconomic status. Both the controls and those with bronchial asthma underwent an assessment of their oral health condition measuring caries experience in both primary and permanent teeth using dmft/DMFT. RESULTS: At the ages of 10, 11 and 12 years a statistically significant higher average dmft values were observed in the children suffering from bronchial asthma as compared with their peers in the control group. In permanent dentition, the DMFT figures were significantly higher at 13 years of age with a value of 6.76 observed in asthmatic subjects compared with 5.06 in the controls. CONCLUSION: Children suffering from bronchial asthma appear to be at higher caries risk and should be provided with particularly intensive preventive dental programmes. PMID- 17078738 TI - Two cases of familial hypomagnesemia with hypercalciuria and nephrocalcinosis: dental findings. AB - BACKGROUND: The authors describe dental and periodontal conditions of two Chinese sisters affected by familial hypomagnesemia with hypercalciuria and nefrocalcinosis (FHHNC). FHHNC is a rare syndrome, genetically investigated since 1999, transmitted as an autosomal recessive disease. It is related to a mutation of PCN-1 gene which encodes for a tight junction protein named paracellin. CASE REPORT: Oral examination of two Chinese patients showed marked enamel hypoplasia, acute gingivitis and periodontal bone loss and severe malocclusion. The literature does not report previous investigations about the dental conditions of FHHNC patients. The syndrome is not definitely described from the clinical point of view. CONCLUSION: Further researches are necessary to understand the linkage between bone loss and enamel structure anomalies in FHHNC and to discover the relationships between nefrocalcinosis, hypomagnesemia, hypercalciuria and amelogenesis imperfecta. PMID- 17078740 TI - Questions applications and conclusions in equine herpesvirus study. PMID- 17078741 TI - Influence of halothane, isoflurane, and sevoflurane on gastroesophageal reflux during anesthesia in dogs. AB - OBJECTIVE: To determine whether maintenance of anesthesia with halothane or sevoflurane is associated with a lower incidence of gastroesophageal reflux (GER) than the use of isoflurane in dogs undergoing orthopedic surgery. ANIMALS: 90 dogs. PROCEDURES: Dogs were evaluated during elective orthopedic surgery. Dogs with a history of vomiting or that had received any drugs that would alter gastrointestinal tract function were excluded from the study. The anesthetic protocol used was standardized to include administration of acepromazine maleate and morphine prior to induction of anesthesia with thiopental. Dogs were allocated to receive halothane, isoflurane, or sevoflurane to maintain anesthesia. A sensor-tipped catheter was placed to measure esophageal pH during anesthesia. Gastroesophageal reflux was defined as an esophageal pH < 4 or > 7.5. RESULTS: 51 dogs had 1 or more episodes of acidic GER during anesthesia. Reflux was detected in 14 dogs receiving isoflurane, 19 dogs receiving halothane, and 18 dogs receiving sevoflurane. In dogs with GER, mean +/- SD time from probe placement to onset of GER was 36 +/- 65 minutes and esophageal pH remained < 4 for a mean of 64% of the measurement period. There was no significant association between GER and start of surgery or moving a dog on or off the surgery table. Dogs that developed GER soon after induction of anesthesia were more likely to regurgitate. CONCLUSIONS AND CLINICAL RELEVANCE: Maintenance of anesthesia with any of the 3 commonly used inhalant agents is associated with a similar risk for development of GER in dogs. PMID- 17078742 TI - Validation of a structured questionnaire as an instrument to measure chronic pain in dogs on the basis of effects on health-related quality of life. AB - OBJECTIVE: To validate the use of a novel questionnaire as an instrument for measurement of chronic pain in dogs through its impact on health-related quality of life (HRQL). ANIMALS: 108 dogs with chronic degenerative joint disease and 26 healthy dogs. PROCEDURES: Questionnaire responses were subjected to factor analysis (FA) and questionnaire scores to discriminant analysis to evaluate construct validity. Questionnaire scores were used to explore the potential of this instrument for minimizing respondent bias and for evaluative purposes. RESULTS: FA results revealed a sensible factor structure accounting for 65% of the variance in data, with factors identifiable as domains of HRQL in dogs affected by chronic pain. Further evidence for construct validity was provided when questionnaire scores were used to discriminate, on the basis of 218 questionnaires, between dogs with clinician-awarded pain scores of 0 and dogs with pain scores >or= 1 (88% discrimination, with 95% of no-pain group dogs and 87% of some-pain group dogs correctly categorized). Use of the questionnaire provided minimized respondent bias. CONCLUSIONS AND CLINICAL RELEVANCE: Validation of the questionnaire as an instrument for discriminative and evaluative measurements of orthopedic chronic pain through its impact on HRQL in dogs was provided. Use of the questionnaire, with further testing and refinement, may support improved clinical decision making, facilitate development of evidence based therapeutic options for chronic diseases, and help veterinarians and owners define humane end points in dogs. IMPACT FOR HUMAN MEDICINE: Information gained here may provide improved measurements of clinical change in animal studies that use dogs with naturally occurring chronic pain to evaluate novel human treatment protocols. PMID- 17078743 TI - Epidemiologic cutoff values for antimicrobial agents against Aeromonas salmonicida isolates determined by frequency distributions of minimal inhibitory concentration and diameter of zone of inhibition data. AB - OBJECTIVE: To develop epidemiologic cutoff values by use of frequency distributions for susceptibility to 4 antimicrobial agents when tested against a representative population of a major aquaculture pathogen, Aeromonas salmonicida. SAMPLE POPULATION: 217 typical and atypical A salmonicida isolates obtained from 20 states and 12 countries. PROCEDURES: Species identification of A salmonicida isolates was confirmed by detection of specific nucleotide sequences by use of a PCR assay. Minimal inhibitory concentration (MIC) and diameter of the zone of inhibition for oxytetracycline, ormetoprim-sulfadimethoxine, oxolinic acid, and florfenicol were determined for each isolate in accordance with standardized antimicrobial susceptibility testing methods that have been approved by the Clinical and Laboratory Standards Institute for bacterial isolates from aquatic animals. Susceptibility data were tabulated in a scattergram and analyzed by use of error rate bounding. RESULTS: Susceptibility tests for oxytetracycline, ormetoprim-sulfadimethoxine, and oxolinic acid revealed 2 distinct populations of bacteria. Isolates tested against florfenicol clustered into a single population. Oxolinic acid susceptibility data revealed higher MICs in the non-United States A salmonicida isolates. Slow-growing (atypical) A salmonicida isolates were generally more susceptible than typical isolates for all antimicrobials, except oxolinic acid. CONCLUSIONS AND CLINICAL RELEVANCE: Use of frequency distributions of susceptibility results to develop epidemiologic cutoff values appears to be applicable to aquatic isolates. Frequency distributions of susceptibility results for A salmonicida revealed clear divisions between isolate susceptibilities. This type of data, considered in conjunction with pharmacokinetic and efficacy data, may be useful for developing clinical breakpoints for use in aquaculture. PMID- 17078744 TI - Effect of width of disk fenestration and a ventral slot on biomechanics of the canine C5-C6 vertebral motion unit. AB - OBJECTIVE: To investigate the effects of disk fenestration and ventral slot formation on vertebral motion unit (VMU) range of motion (ROM) and determine the effects of fenestration and ventral slot width on VMU ROM. SAMPLE POPULATION: C5 C6 VMUs from 10 skeletally mature canine cadavers. PROCEDURES: Specimens were assigned to 2 groups (5 specimens/group). Surgery was performed in which width of a fenestration and a ventral slot was 33% (group 1) or 50% (group 2) the width of the vertebral body. Flexion-extension, lateral bending, and axial torsion ROMs were measured during loading before surgery, after fenestration, and after ventral slot formation. Range of motion was compared within groups to determine effects of surgical procedure on stability and between groups to determine effects of width of fenestration and ventral slot on stability. RESULTS: For both groups, fenestration resulted in a significant increase in ROM during flexion extension, compared with results for intact specimens. Ventral slot formation resulted in a significant increase in ROM during flexion-extension and lateral bending, compared with results for intact specimens. Ventral slot formation resulted in a significant increase in ROM only during flexion-extension, compared with results for fenestrated specimens. There were no significant differences in ROM of the intact, fenestrated, and ventral slot specimens between groups. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of these results suggests that fenestration and ventral slot procedures each affect the biomechanics of the C5 C6 VMU. Width of a fenestration or ventral slot up to 50% of the width of C5-C6 may be clinically acceptable. PMID- 17078745 TI - Inclination of the patellar ligament in relation to flexion angle in stifle joints of dogs without degenerative joint disease. AB - OBJECTIVE: To measure the angles between the patellar ligament and the tibial plateau and between the patellar ligament and the common tangent at the tibiofemoral contact point (TFCP) throughout the full range of motion of the stifle joint in dogs and determine the flexion angles at which the patellar ligament is perpendicular to the tibial plateau or to the common tangent. SAMPLE POPULATION: 16 hind limbs from cadavers of 9 adult dogs without radiographically detectable degenerative joint disease. PROCEDURES: Mediolateral radiographic views of the stifle joints from full extension through full flexion were obtained (10 degrees increments). Angles between the tibial and femoral long axes (beta), between the patellar ligament and the tibial plateau gamma), and between the patellar ligament and the common tangent at TFCP (alpha) were measured. Data were analyzed via simple linear regression. RESULTS: In canine stifle joints, angles gamma and alpha decreased linearly with increasing flexion (angle beta). The patellar ligament was perpendicular to the tibial plateau and perpendicular to the common tangent at the TFCP at 90 degrees and 110 degrees of flexion, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: By use of the conventionally defined tibial plateau, data suggest that at approximately 90 degrees of flexion in stifle joints of dogs, shear force in the sagittal plane exerted on the proximal portion of the tibia shifts the loading from the cranial to the caudal cruciate ligament. Analyses involving the common tangent at the TFCP (a more anatomically representative reference point) identified this crossover point at approximately 110 degrees of joint flexion. PMID- 17078746 TI - Angle between the patellar ligament and tibial plateau in dogs with partial rupture of the cranial cruciate ligament. AB - OBJECTIVE: To measure the angles between the patellar ligament and the tibial plateau and between the patellar ligament and the common tangent at the tibiofemoral contact point (TFCP) in stifle joints of dogs with partial rupture of the cranial cruciate ligament (CrCL) for comparison with data obtained for stifle joints in dogs with intact CrCLs. SAMPLE POPULATION: 60 stifle joints of 54 dogs with surgically confirmed partial CrCL rupture. PROCEDURES: Mediolateral radiographic views of the stifle joints were obtained, and the angles between the patellar ligament and the conventionally defined tibial plateau (angle gamma) and between the patellar ligament and the common tangent to the TFCP (angle alpha) were measured at incidental stifle joint flexion (angle beta) by 2 independent observers. Data underwent linear regression analysis and were compared with findings in joints of dogs without degenerative joint disease. RESULTS: In stifle joints of dogs with a partial rupture of the CrCL, angles gamma and alpha were 5 degrees and 2 degrees larger than each corresponding angle in healthy canine joints. At 100 degrees of flexion, the patellar ligament was perpendicular to the conventionally defined tibial plateau. At 110 degrees of flexion, the patellar ligament was perpendicular to the common tangent at the TFCP. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs, stifle joints with partially ruptured CrCLs have marginally larger angles between the patellar ligament and the tibial plateau, compared with joints with intact CrCLs; at equivalent angles of flexion, comparatively greater shear force affects the CrCLs in stifle joints with partial CrCL ruptures. PMID- 17078747 TI - Differences in hematocrit of blood samples obtained from two venipuncture sites in sharks. AB - OBJECTIVE: To evaluate differences in Hct between 2 venipuncture sites in captive and free-ranging sharks. ANIMALS: 32 healthy adult captive sharks (Carcharhinus melanopterus, Carcharhinus plumbeus, Stegastoma fasciatum, Orectolobus japonicus, and Triaenodon obesus) and 15 captured free-ranging adult sharks (Carcharhinus limbatus and Carcharhinus acronotus). PROCEDURES: Blood samples were collected from the caudal tail artery followed by collection from the sinus located immediately caudal to the cranial dorsal fin. The Hct was determined for each sample and results were compared. Additionally, results for sharks that were highly active and used aerobic metabolism were compared with results for sharks that were less active and tolerant of anaerobic conditions. RESULTS: Mean Hct for all sharks was significantly less (8% less) in blood samples obtained from the cranial dorsal fin sinus, compared with the Hct for samples obtained from the caudal tail artery. When compared on the basis of metabolic class, sharks that were more tolerant of anaerobic conditions had lower Hct values and smaller differences between the 2 venipuncture sites. CONCLUSIONS AND CLINICAL RELEVANCE: Hct values were significantly lower in blood samples collected from the cranial dorsal fin sinus compared with values for samples collected from the caudal tail artery. It is important to recognize this difference when evaluating hematologic variables in sharks and when establishing reference ranges for Hcts for shark populations. Sharks that were more active and relied on aerobic metabolism had higher Hct values than did anaerobic-tolerant sharks, and the difference in Hct values between venipuncture sites was more pronounced. PMID- 17078748 TI - Thin-slice three-dimensional gradient-echo magnetic resonance imaging of the pituitary gland in healthy dogs. AB - OBJECTIVE: To evaluate thin-slice 3-dimensional gradient-echo (GE) magnetic resonance imaging (MRI) of the pituitary gland in healthy dogs. ANIMALS: 11 healthy dogs. PROCEDURES: By use of a 0.2-Tesla open magnet, MRI of the skull was performed with T1-weighted GE sequences and various protocols with variations in imaging plane, slice thickness, and flip angle before and after administration of contrast medium; multiplanar reconstructions were made. The pituitary region was subjectively assessed, and its dimensions were measured. Image quality was determined by calculation of contrast-to-noise and signal-to-noise ratios. RESULTS: Best-detailed images were obtained with a T1-weighted GE sequence with 1 mm slice thickness and 30 degrees flip angle before and after administration of contrast medium. Images with flip angles > 50 degrees were of poor quality. Quality of multiplanar reconstruction images with 1-mm slices was better than with 2-mm slices. The bright signal was best seen without contrast medium. With contrast medium, the dorsal border of the pituitary gland was clearly delineated, but lateral borders were more difficult to discern. CONCLUSIONS AND CLINICAL RELEVANCE: MRI of the canine pituitary gland with a 0.2-Tesla open magnet should include a T1-weighted GE sequence with 1-mm slice thickness and flip angle of 30 degrees before and after administration of contrast medium. The neurohypophysis was best visualized without contrast medium. The MRI examination permitted differentiation between the pituitary gland and surrounding structures. PMID- 17078749 TI - In vitro effects of hydrochloric acid and various concentrations of acetic, propionic, butyric, or valeric acids on bioelectric properties of equine gastric squamous mucosa. AB - OBJECTIVE: To compare the effects of hydrochloric acid (HCl) and various concentrations of volatile fatty acids (VFAs) on tissue bioelectric properties of equine stomach nonglandular (NG) mucosa. SAMPLE POPULATION: Gastric tissues obtained from 48 adult horses. PROCEDURES: NG gastric mucosa was studied by use of Ussing chambers. Short-circuit current (Isc) and potential difference (PD) were measured and electrical resistance (R) and conductance calculated for tissues after addition of HCl and VFAs (5, 10, 20, and 40 mM) in normal Ringer's solution (NRS). RESULTS: Mucosa exposed to HCl in NRS (pH of 1.5 and, to a lesser extent, 4.0) had a significant decrease in Isc, PD, and R, whereas tissues exposed to acetic acid at a pH of < 4.0, propionic and butyric acids at a pH of 4 weeks old to adulthood. CONCLUSIONS AND CLINICAL RELEVANCE: Findings suggested that colonization by a swine-origin HPLO, H pylori, or both and resultant seroconversion, like that of H pylori infection in humans, were common in commercial swine operations. Furthermore, data indicated that gastric infection was acquired at an early age. The relationships to gastric colonization by HPLOs and clinical manifestations of disease such as gastritis and gastroesophageal ulceration remain to be determined. PMID- 17078752 TI - Assessment of IgE binding to native and hydrolyzed soy protein in serum obtained from dogs with experimentally induced soy protein hypersensitivity. AB - OBJECTIVE: To assess binding of IgE to native, whole hydrolyzed, and separated hydrolyzed fractions of soy protein in serum obtained from dogs with experimentally induced soy protein hypersensitivity. ANIMALS: 8 naive Beagles (6 experimentally sensitized to native soy protein and 2 control dogs). PROCEDURES: 6 dogs were sensitized against soy protein by administration of allergens during a 90-day period. After the sensitization protocol was completed, serum concentrations of soy-specific IgE were measured and intradermal skin tests were performed in all 6 dogs to confirm that the dogs were sensitized against soy protein. Serum samples from each sensitized and control dog underwent western blot analysis to assess the molecular mass band pattern of the different allergenic soy fractions and evaluate reactivities to native and hydrolyzed soy protein. RESULTS: In sera from sensitized dogs, a characteristic band pattern with 2 major bands (approx 75 and 50 kd) and 2 minor bands (approx 31 and 20 kd) was detected, whereas only a diffuse band pattern associated with whole hydrolyzed soy protein was detected in the most reactive dog. Reactivity was evident only for the higher molecular mass peptide fraction. In control dogs, no IgE reaction to native or hydrolyzed soy protein was detected. CONCLUSIONS AND CLINICAL RELEVANCE: Data suggest that the binding of soy-specific IgE to the hydrolyzed soy protein used in the study was significantly reduced, compared with binding of soy-specific IgE to the native soy protein, in dogs with experimentally induced soy hypersensitivity. PMID- 17078753 TI - Evaluation of the effects of short-chain fatty acids and extracellular pH on bovine neutrophil function in vitro. AB - OBJECTIVE: To investigate the effects of short-chain fatty acids (SCFAs) and pH on neutrophil oxidative burst, phagocytosis, and morphology after exposure to acetate, propionate, butyrate, or succinate at pH 5.5 and 6.7. SAMPLE POPULATION: Neutrophils isolated from bovine blood samples and Porphyromonas levii, Prevotella spp, and Bacteroides fragilis isolated from lesions of cattle with acute interdigital phlegmon (foot rot). PROCEDURES: Bacteria were cultured in strictly anaerobic conditions. Bacterial SCFA production was measured with high performance liquid chromatography. Neutrophils were isolated, stimulated with phorbol 12-myristate 13-acetate (PMA) or opsonized zymosan (OZ), and incubated with dihydroethidium or dichlorofluorescein diacetate to measure production of O(2)and H(2)O(2), respectively. Phagocytosis was assessed after exposure to serum opsonized bacteria. Cellular morphology was assessed with differential staining. RESULTS: All bacteria produced at least 3 of the 4 SCFAs. Production of both O(2) and H(2)O(2) was markedly curtailed in PMA-stimulated neutrophils exposed to SCFA at pH 5.5, compared with production at pH 6.7. Succinate caused a significant dose-dependent decrease in O(2) production at pH 6.7 in OZ-stimulated neutrophils. Monoprotic SCFAs elicited a significant increase in H(2)O(2) production in OZ-stimulated neutrophils at pH 6.7 but a significant decrease at pH 5.5. Monoprotic SCFAs significantly increased phagocytosis at pH 6.7 but decreased phagocytic activity at pH 5.5. Cellular necrosis was observed in cells exposed to SCFAs at pH 5.5. CONCLUSIONS AND CLINICAL RELEVANCE: Establishment and persistence of anaerobic bacteria in cattle with foot rot infection may result in part from neutrophil dysfunction secondary to the effects of bacterially secreted SCFA in acidotic microenvironments. PMID- 17078754 TI - Establishment of optimal conditions for long-term culture of erythrocytic stages of Theileria uilenbergi. AB - OBJECTIVE: To establish optimal conditions for long-term culture of the erythrocytic stage of Theileria uilenbergi. SAMPLE POPULATION: Red blood cells from 3 splenectomized sheep experimentally infected with a blood stabilate of T uilenbergi. PROCEDURES: Cultures of T uilenbergi were initiated by use of blood from experimentally infected sheep collected when parasites were detected in Giemsa-stained thin blood smears. Different culture conditions were tested to optimize in vitro growth of the organisms. Subcultures were performed at a ratio of 1:2, 1:4, and 1:8 when the percentage of parasitized erythrocytes (PPE) was at least 1% or when the initial PPE was doubled. RESULTS: The optimal culture medium was HL-1 medium (a complete chemically defined medium) supplemented with 20% sheep serum and 0.75% chemically defined lipids. Optimal culture conditions included incubation in a humidified 2% O(2), 5% CO(2), and 93% N(2) atmosphere at 37 degrees C. Cultures of the merozoite stage of the parasite were continuously propagated in vitro for > 1 year. The PPE reached values of up to 3%. CONCLUSIONS AND CLINICAL RELEVANCE: Optimization of culture conditions to reach a high PPE seems worthwhile. The continuous propagation of T uilenbergi in culture allows the production of parasite material without infecting animals and provides a continuous laboratory source of parasites for further studies. PMID- 17078755 TI - Evaluation of a model for Escherichia coli O157:H7 colonization in streptomycin treated adult cattle. AB - OBJECTIVE: To develop a repeatable model for studying colonization with streptomycin-resistant Escherichia coli O157:H7 in adult cattle. ANIMALS: 5 adult mixed-breed beef cattle. PROCEDURES: Cattle were surgically cannulated in the duodenum, treated daily with streptomycin (33 mg/kg) via the duodenal cannula prior to and during experimental colonizations, and colonized with 10(10) CFUs of streptomycin-resistant E coli O157:H7 via the duodenal cannula. Colonization of rectal mucus and shedding in feces were monitored. Antimicrobials were administered to eliminate the colonizing strain so that 5 repeated colonization experiments could be performed. A comprehensive analysis of colonization was performed at necropsy. RESULTS: Streptomycin treatment resulted in improved experimental colonization variables, compared with untreated controls, during initiation (days 2 to 6) and early maintenance (days 7 to 12) of colonization. Elimination of the colonizing strain followed by 5 repeated colonizations in the same animals indicated the repeatability of the protocol. Positive results of bacteriologic culture of feces 7 and 12 days after colonization were obtained in 100% and 84% of samples, respectively, across all animals and trials. At necropsy, highest magnitude recovery was in terminal rectal mucus. CONCLUSIONS AND CLINICAL RELEVANCE: The model was highly repeatable and novel with respect to streptomycin treatment, use of duodenal cannulas, and repeated colonizations of the same animals. Its use in adult cattle, from which most bovine-derived food originates, is critical to the study of preharvest food safety. The findings have implications for understanding intermittency of shedding in the field and for proposed vaccine-based interventions. PMID- 17078756 TI - Expression of neural markers on bone marrow-derived canine mesenchymal stem cells. AB - OBJECTIVE: To evaluate cell surface markers of bone marrow-derived canine mesenchymal stem cells (MSCs) by use of flow cytometric analysis and determine whether canine MSCs express proteins specific to neuronal and glial cells. SAMPLE POPULATION: Bone marrow aspirates collected from iliac crests of 5 cadavers of young adult dogs. PROCEDURES: Flow cytometric analysis was performed to evaluate cell surface markers and homogeneity of third-passage MSCs. Neural differentiation of canine MSCs was induced by use of dibutyryl cAMP and methyl isobutylxanthine. Expressions of neuronal (beta III-tubulin) and glial (glial fibrillary acidic protein [GFAP] and myelin basic protein) proteins were evaluated by use of immunocytochemical and western blot analyses before and after neural differentiation. RESULTS: Third-passage canine MSCs appeared morphologically homogeneous and shared phenotypic characteristics with human and rodent MSCs. Immunocytochemical and western blot analyses revealed that canine MSCs constitutively expressed beta III-tubulin and GFAP. After induction of neural differentiation, increased expression of GFAP was found in all samples, whereas such change was inconsistent in beta III-tubulin expression. Myelin basic protein remained undetectable on canine MSCs for these culture conditions. CONCLUSIONS AND CLINICAL RELEVANCE: Canine bone marrow-derived mononuclear cells yielded an apparently homogeneous population of MSCs after expansion in culture. Expanded canine MSCs constitutively expressed neuron or astrocyte specific proteins. Furthermore, increases of intracellular cAMP concentrations induced increased expression of GFAP on canine MSCs, which suggests that these cells may have the capacity to respond to external signals. Canine MSCs may hold therapeutic potential for treatment of dogs with neurologic disorders. PMID- 17078757 TI - Effects of the bronchoalveolar lavage procedure on lung function in horses with clinical exacerbation of recurrent airway obstruction. AB - OBJECTIVE: To evaluate whether bronchoalveolar lavage (BAL) alters respiratory mechanics of horses with recurrent airway obstruction (ie, heaves) over a 48-hour period. ANIMALS: 6 horses affected with heaves. PROCEDURES: Horses were subjected to a complete BAL procedure, which included sedation with xylazine and butorphanol, intratracheal administration of lidocaine, and instillation and aspiration of two 250-mL boluses of saline (0.9% NaCl) solution through an endoscope (study 1). To evaluate the effects of saline solution, horses were subjected to the same procedure without saline solution instillation and aspiration (study 2). Lastly, the endoscope was similarly introduced into the lower airways, without sedation or saline instillation and aspiration (study 3). Respiratory mechanics were performed at baseline (time 0) and at 3, 6, 12, 24, and 48 hours after each procedure. RESULTS: In study 1, BAL induced a significant decrease in pulmonary resistance lasting up to 6 hours. This may have resulted from clearance of mucus in large airways. We also observed a significant increase in lung elastance and transpulmonary pressure at 12 hours after BAL in all 3 studies, which may be attributed to a circadian effect. CONCLUSIONS AND CLINICAL RELEVANCE: Our results indicate that the temporal effects of BAL procedures on lung mechanics should be taken into account when designing research protocols involving horses with heaves. Future studies should address the immediate effects of BAL on lung function. PMID- 17078758 TI - Results of vaccination of Asian elephants (Elephas maximus) with monovalent inactivated rabies vaccine. AB - OBJECTIVE: To evaluate the humoral immune response of Asian elephants to a primary IM vaccination with either 1 or 2 doses of a commercially available inactivated rabies virus vaccine and evaluate the anamnestic response to a 1-dose booster vaccination. ANIMALS: 16 captive Asian elephants. PROCEDURES: Elephants with no known prior rabies vaccinations were assigned into 2 treatment groups of 8 elephants; 1 group received 1 dose of vaccine, and the other group received 2 doses of vaccine 9 days apart. All elephants received one or two 4-mL IM injections of a monovalent inactivated rabies virus vaccine. Blood was collected prior to vaccination (day 0) and on days 9, 35, 112, and 344. All elephants received 1 booster dose of vaccine on day 344, and a final blood sample was taken 40 days later (day 384). Serum was tested for rabies virus-neutralizing antibodies by use of the rapid fluorescent focus inhibition test. RESULTS: All elephants were seronegative prior to vaccination. There were significant differences in the rabies geometric mean titers between the 2 elephant groups at days 35, 112, and 202. Both groups had a strong anamnestic response 40 days after the booster given at day 344. CONCLUSIONS AND CLINICAL RELEVANCE: Results confirmed the ability of Asian elephants to develop a humoral immune response after vaccination with a commercially available monovalent inactivated rabies virus vaccine and the feasibility of instituting a rabies virus vaccination program for elephants that are in frequent contact with humans. A 2-dose series of rabies virus vaccine should provide an adequate antibody response in elephants, and annual boosters should maintain the antibody response in this species. PMID- 17078761 TI - Networking at the second Interactome Meeting. PMID- 17078759 TI - Proteins are specifically altered in disease--really? PMID- 17078762 TI - Clinical proteomics: an insight into the urinary proteome. Interview with Dr. Hassan Dihazi. PMID- 17078763 TI - Proteomic profiling of differentiating osteoblasts. AB - The major event that triggers osteogenesis is the transition of mesenchymal stem cells into bone-forming, differentiating osteoblast cells. Osteoblast differentiation is the primary event of bone formation, exemplified by the synthesis, deposition and mineralization of extracellular matrix. Osteoblast differentiation is controlled tightly by sequential activation of diverse transcription factors that regulate the expression of specific genes. The spatial and temporal regulation of the differentiation process is not completely understood at the cellular or molecular level. Recent advances in mass spectrometry-based proteomics have allowed for the systematic qualitative and quantitative profiling of differentiating osteoblasts, enabling a better understanding of the multiple factors and signaling events that control the differentiation process at a molecular level. This review focuses on recent developments in the proteomic analysis of differentiating osteoblasts, including advances, challenges and future prospects of using mass spectrometry to investigate the local and systemic factors regulating bone formation and its homeostasis. PMID- 17078764 TI - Developing a tool for noninvasive monitoring of renal allografts. AB - Renal transplantation has emerged as the therapy of choice for many patients with end-stage renal disease. One of the major goals is to tailor immunosuppressive therapy to the individual needs of every patient at every time point post transplant, balancing the risk for rejection and over-immunosuppression. Such individualized treatment will require assays that can detect harmful processes in the allograft early and that can be measured repeatedly. In this review, advantages and disadvantages of current assays to monitor renal allografts noninvasively and how proteomic technology might contribute to the development of novel biomarkers to improve patient management will be discussed. PMID- 17078765 TI - Nonantibody-based recognition: alternative molecules for detection of pathogens. AB - Immunoassays have been well established for many years as the cornerstone of detection technologies. These assays are sensitive, selective and, in general, highly resistant to interference from complex sample matrices when compared with nucleic acid-based tests. However, both antibody- and nucleic acid-based detection systems require a priori knowledge of the target and development of specific reagents; multiplexed assays can become increasingly problematic when attempting to detect a plethora of different targets, the identities of which are unknown. In an effort to circumvent many of the limitations inherent in these conventional assays, other recognition reagents are being explored as alternatives, or indeed as adjuncts, to antibodies for pathogen and toxin detection. This article will review a number of different recognition systems ranging in complexity from small molecules, such as nucleic-acid aptamers, carbohydrates and peptides, to systems as highly complicated as whole cells and organisms. All of these alternative systems have tremendous potential to achieve superior sensitivity, selectivity, and stability, but are also subject to their own limitations, which are also discussed. In short, while in its infancy, this field holds great promise for the development of rapid, fieldable assays that are highly complementary to existing antibody- and nucleic acid-based technologies. PMID- 17078766 TI - Resonance Raman spectroscopy in malaria research. AB - In recent years, the field of Raman spectroscopy has witnessed a surge in technological development, with the incorporation of ultrasensitive, charge coupled devices, improved laser sources and precision Rayleigh-filter systems. This has led to the development of sensitive confocal micro-Raman spectrometers and imaging spectrometers that are capable of obtaining high spatial-resolution spectra and images of subcellular components within single living cells. This review reports on the application of resonance micro-Raman spectroscopy to the study of malaria pigment (hemozoin), a by-product of hemoglobin catabolization by the malaria parasite, which is an important target site for antimalarial drugs. The review aims to briefly describe recent studies on the application of this technology, elucidate molecular and electronic properties of the malaria pigment and its synthetic analog beta-hematin, provide insight into the mechanism of hemozoin formation within the food vacuole of the parasite, and comment on developing strategies for using this technology in drug-screening protocols. PMID- 17078767 TI - Understanding the folding of GFP using biophysical techniques. AB - Green fluorescent protein (GFP) and its many variants are probably the most widely used proteins in medical and biological research, having been extensively engineered to act as markers of gene expression and protein localization, indicators of protein-protein interactions and biosensors. GFP first folds, before it can undergo an autocatalytic cyclization and oxidation reaction to form the chromophore, and in many applications the folding efficiency of GFP is known to limit its use. Here, we review the recent literature on protein engineering studies that have improved the folding properties of GFP. In addition, we discuss in detail the biophysical work on the folding of GFP that is beginning to reveal how this large and complex structure forms. PMID- 17078768 TI - The relations between psychosocial factors at work and health status among workers in home care organizations. AB - A considerable proportion of sickness absence and disability pension is caused by subjective health complaints, especially low back pain (LBP). In recent years focus has been on psychosocial characteristics of work as potential risk factors. The aim of this study is to examine the relations between psychosocial work aspects and subjective health complaints, LBP, and need for recovery. A total of 779 employees working in home care participated in a cross-sectional study. Higher psychological demands were associated with subjective health complaints and need for recovery. However, decision authority, skill discretion, and the 2 aspects of social support did not seem to be important factors. Higher psychological demands do not seem to be associated with severity of LBP, but does show an association with sick leave. PMID- 17078769 TI - Diurnal saliva cortisol levels and relations to psychosocial factors in a population sample of middle-aged Swedish men and women. AB - Whereas psychosocial risk factors increase the risk for disease, psychosocial resources reduce this risk. To examine a possible pathway for these effects, the relations between saliva cortisol levels and psychosocial factors were studied in a random sample of 257 men and women aged 30 to 64 years. Saliva samples were collected at home on waking, 30 min after waking, and in the evening. A flatter diurnal rhythm of cortisol, that is, lower deviations between awakening and evening cortisol levels, was related to high levels of psychosocial risk factors (cynicism, depression, and vital exhaustion), whereas a steeper diurnal rhythm was related to psychosocial resources (social support and coping), general health, and well-being (all p < .05). Our results support earlier suggestions that the capacity of the hypothalamic-pituitary-adrenal-axis to dynamically respond to stress is 1 pathway for observed effects of psychosocial factors regarding risk for disease development. PMID- 17078770 TI - Heart rate variability and intima media thickness. AB - Increased activity in the sympathetic nervous system is part of the physiological stress response and is expressed in the heart rate variability (HRV). The objective of this study was to examine associations of HRV and intima media thickness (IMT). In 2002, satisfactory measurements of HRV of 78 voluntary participants were made, both during a stress test and during sleep. IMT in 2002 and the progression in IMT from 1998 to 2002 were analyzed in relation to measures of HRV. HRV was negatively associated with IMT and IMT progression both during test and sleep. In men with higher IMT measures than mean the low frequency-high frequency ratio was higher during sleep than during test, perhaps mirroring a lack of ability to recovery. HRV was negatively associated with IMT and IMT progression. HRV may be part of the pathophysiological pathway between psychological strain and atherosclerosis. PMID- 17078771 TI - Effects of fasting therapy on irritable bowel syndrome. AB - How to treat patients with irritable bowel syndrome (IBS) who do not respond to pharmacotherapy is an unsolved problem. Psychotherapy, which has been reported on in previous studies, is available only in specific centers. We describe in this study a novel and simple psychotherapy; that is, the fasting therapy (FT) for treatment of patients with IBS. Of 84 inpatients with IBS, 58 patients who still had moderate to severe IBS symptoms after 4-week basic treatment were investigated retrospectively. Of the 58 patients enrolled in this study, 36 underwent FT, whereas the remaining 22 received a consecutive basic treatment (control therapy). There were no significant differences in the 4-point severity scales of gastrointestinal and psychological symptoms between the 2 groups before the start of FT. The basic treatment consisted of pharmacotherapy and brief psychotherapy, whereas the FT consisted of 10 days of starvation followed by 5 days of refeeding. Changes in scores of symptoms before and after each treatment were analyzed. FT significantly improved 7 out of the 10 symptoms assessed; that is, abdominal pain-discomfort (p < .001), abdominal distension (p < .001), diarrhea (p < .001), anorexia (p = .02), nausea (p < .01), anxiety (p < .001), and interference with life in general (p < .001). However, the control therapy significantly improved only 3 out of the 10 symptoms assessed; that is, abdominal pain-discomfort (p = .03), abdominal distension (p < .01), and interference with life (p = .01). Our results suggest that FT may have beneficial effects on intractable patients with IBS. PMID- 17078773 TI - Perceived control and change in physical functioning after coronary artery bypass grafting: a prospective study. AB - We determined if perceived general and health-related control predicted change in physical functioning after coronary artery bypass grafting (CABG) in 1,038 patients. Control and physical functioning were assessed prior to hospital discharge and physical functioning also was assessed 6 weeks and 6 months later. The repeated measures outcome was modeled using a one-way analysis of covariance with interaction terms for General and Health-Related Control x Time. The sample was primarily composed of Caucasian (94%) men (73%) with a mean age of 65 +/- 11 years. Increasing health-related control was an independent predictor of positive change in physical functioning with a stronger effect in women. There was a nearly significant association between increasing general control and positive physical functioning change. Lack of significant interactions with time indicate that higher control has a beneficial impact on physical functioning change at 6 weeks and 6 months. Assessing control may help clinicians predict post-CABG physical functioning and may shape intervention efforts. PMID- 17078772 TI - The effects of distal limb warming on sleep latency. AB - Sleep onset is best predicted by an increase in the amount of hand and foot warming relative to more proximal areas (Krauchi, Cajochen, Werth, & Wirz Justice, 1999; Krauchi, Cajochen,Werth,& Wirz-Justice, 2000).We assessed if an increase in sleepiness would also occur if the distal limbs were warmed through an external manipulation. In this study, 5 min before participants (N = 11) attempted to fall asleep on a multiple sleep latency test (Carskadon et al., 1986) their hands and feet were immersed in water heated to either 42 degrees C or heated to the temperature of the warmest limbs. The results show no difference in sleep latency between the warm and control water conditions. There was a decrease in sleep latency in the control and warm water conditions compared to the initial (noncounterbalanced) baseline multiple sleep latency test. Further studies are necessary to determine if any degree of limb warming hastens sleep onset. PMID- 17078774 TI - Cognitive, behavioral, and physiological reactivity to chronic itching: analogies to chronic pain. AB - It is well-known that cognitive, behavioral, and physiological reactivity to pain, such as catastrophizing, avoidance of activity, and increased physiological responses, can unfavorably affect long-term outcomes in patients with chronic pain. In line with similarities between the psychophysiology of pain and itching, corresponding mechanisms may be relevant for the maintenance of chronic itching. The goal of this study was to examine the role of self-reported cognitive, behavioral, and physiological reactivity factors on itching-related outcomes in 235 patients with chronic skin diseases suffering from chronic itching. Sequential regression analyses indicate that all 3 reactivity systems predicted itching-related outcomes. Specifically, more catastrophizing, higher levels of avoidance of activity, and heightened self-reported physiological reactivity predicted more itching, more scratching, and a reduced disease-related quality of life. The results suggest that a psychological model as described for chronic pain is a useful starting point for study of the maintaining mechanisms of chronic itching. PMID- 17078775 TI - Differential diagnosis of chronic fatigue syndrome and major depressive disorder. AB - The goal of this study was to identify variables that successfully differentiated patients with chronic fatigue syndrome, major depressive disorder, and controls. Fifteen participants were recruited for each of these three groups, and discriminant function analyses were conducted. Using symptom occurrence and severity data from the Fukuda et al. (1994) definitional criteria, the best predictors were postexertional malaise, unrefreshing sleep, and impaired memory concentration. Symptom occurrence variables only correctly classified 84.4% of cases, whereas 91.1% were correctly classified when using symptom severity ratings. Finally, when using percentage of time fatigue reported, postexertional malaise severity, unrefreshing sleep severity, confusion-disorientation severity, shortness of breath severity, and self-reproach to predict group membership, 100% were classified correctly. PMID- 17078777 TI - Exercise beliefs of breast cancer survivors before and after participation in a randomized controlled trial. AB - The purpose of this study was to examine the exercise beliefs of breast cancer survivors before and after participation in a randomized trial. Prior to randomization, 52 breast cancer survivors completed exercise belief measures based on the theory of planned behavior. After the trial, participants assigned to the exercise group (n = 24) completed the belief measures again. Results show that there was significant variability in the expected benefits of exercise prerandomization, ranging from 40% for a reduced risk of breast cancer recurrence to 94% for an improved energy level. Moreover, attitudes toward exercise and perceptions of control were higher in the exercise group after the exercise program. The findings are discussed in terms of the veracity of the exercise beliefs held by breast cancer survivors as well as the aspects of the program that may have contributed to the positive changes in exercise beliefs. PMID- 17078776 TI - Fatigue varies by social class in African Americans but not Caucasian Americans. AB - Socioeconomic status explains many ethnic disparities in health; however, mechanisms are hard to identify. Fatigue-a frequent complaint in patients and normals-is associated with poorer quality of life. We wondered if ethnicity and social class interact to explain fatigue. A total of 40 African Americans (AAs) and 64 Caucasian Americans (CAs) completed short forms of the Profile of Mood States (POMS-SF) and Multidimensional Fatigue Symptom Inventory (MFSI-SF). Participants were divided into high-middle and low social class groups (as per Hollingshead, 1958a). After controlling for gender, body mass index, depressive symptoms, and response bias, ethnicity and social class interacted for POMS-SF fatigue. AAs in the high-middle classes reported more fatigue than AAs in the low classes and CAs in the high-middle classes. Fatigue did not differ by class for CAs nor by ethnicity in the lower classes. Similar findings emerged for MFSI-SF general fatigue. Social class is important for understanding fatigue in AAs but not CAs. PMID- 17078779 TI - Aggression in attention-deficit/hyperactivity disorder children: diagnostic or comorbid symptom? PMID- 17078778 TI - Does timing and sequencing of transitions to adulthood make a difference? Stress, smoking, and physical activity among young Australian women. AB - The major changes of the transition to adulthood are argued to be stressful, and health-related behaviors such as smoking and physical activity may be adopted, consolidated, or abandoned at this time. On the other hand, research has suggested that the normative transitions of emerging adulthood, although involving considerable change, may be associated with low stress because they are perceived as both positive and normal at this life stage. This article examines relations between the timing and sequencing of life transitions and stress and health-related behaviors, focusing on the transition to young adulthood among Australian women. A total of 853 women aged 22 to 27 provided information about the timing and sequencing of 6 life transitions: moving out of home, stopping full-time education, starting full-time work, having the first live-in relationship, marriage, and motherhood-and stress, smoking, and physical activity. Most had moved out of home, stopped full-time education, and started full-time work, but only 14% had undertaken all 6 transitions. Overall, 70% of participants had made transitions "in order." Overall, the findings suggest that the relations between timing and sequencing of transitions, and indicators of health, are moderate for smoking, but small for stress and for physical activity. These effects remained after controlling for socioeconomic status of the participants' families of origin. Matching current social norms for the timing and sequencing of life changes may be of less importance for women's well-being than is commonly believed. Although the significant relations between early or "out of order" transitions and smoking are of concern, the smaller relations with stress and with sedentariness suggest that such transitions may have limited negative consequences, and support the view that individuals are active in choosing the life path that is appropriate for them and their circumstances. PMID- 17078781 TI - Enteral levodopa/carbidopa gel infusion for the treatment of motor fluctuations and dyskinesias in advanced Parkinson's disease. AB - Pharmacotherapy designed to alleviate the symptoms of Parkinson's disease is focused on the stimulation of striatal dopamine receptors. The ideal pharmacological treatment would involve continuous dopaminergic stimulation. Currently, the most effective pharmacotherapy available is levodopa delivered orally; however, its short plasma half-life in combination with erratic gastric emptying and intestinal absorption makes dopaminergic stimulation pulsatile. Motor fluctuations and dyskinesias develop with time as a consequence and can become disabling. A gel formulation of levodopa/carbidopa has been developed for enteral (duodenal or jejunal) infusion via a portable pump. The infusion provides smooth plasma levodopa levels, more continuous dopaminergic stimulation and effective treatment of motor complications. PMID- 17078782 TI - Eletriptan: a review and new perspectives. AB - Eletriptan is a second-generation 5-hydroxytryptamine(1B/1D) receptor agonist, or triptan, indicated for the acute treatment of migraine. Eletriptan has a favorable pharmacokinetic and pharmacodynamic profile expressed by bioavailability, half-life and high selectivity for cranial arteries. It has been shown to be effective and well tolerated in a wide preapproval development program, which included over 11,000 patients and treated more than 74,000 migraine attacks. In clinical trials, eletriptan has been demonstrated to be one of the most effective oral therapies for the acute treatment of migraine and has shown a very high safety and tolerability profile across the studies performed. Eletriptan showed the most favorable cost-effectiveness profile when compared with other agents in its class. PMID- 17078784 TI - Attention-deficit/hyperactivity disorder: using P300 topography to choose optimal treatment. AB - Attention-deficit/hyperactivity disorder is the most prevalent behavioral disorder in children, and persists into adulthood. Stimulants (methylphenidate and amphetamines) with dopaminergic mechanisms are the most commonly used pharmacological treatment. Nonselective (desipramine and imipramine) and selective (atomoxetine) norepinephrine reuptake inhibitors can also be effective. What constitutes a sufficient response to treatment? Too often a partial response, leaving the patient symptomatic, is accepted. If response is defined more strictly, allowing for a return to normal, then the usually quoted 70% response rates to any given attention-deficit/hyperactivity disorder medicine drop to approximately 40%. With different medicines and not enough patients responding robustly to any given medicine, we can use medicines sequentially to find the medicine that produces a robust response. Alternatively, P300 topography can be used to select optimal treatment. PMID- 17078783 TI - Ziconotide: a new nonopioid intrathecal analgesic for the treatment of chronic pain. AB - Ziconotide is a new nonopioid intrathecal agent recently approved for the treatment of chronic pain. Ziconotide is indicated for the management of severe chronic pain in patients for whom intrathecal therapy is warranted and who are intolerant of or refractory to other treatment, such as systemic analgesics, adjunctive therapies or intrathecal morphine. Ziconotide blocks the N-type calcium channels located in the superficial dorsal horn of the spinal cord, resulting in potent analgesia. The efficacy of ziconotide has been demonstrated in three randomized, placebo-controlled trials in over 500 patients. In addition, its safety has been demonstrated in over 1200 subjects. Ziconotide is a potent analgesic with a narrow therapeutic window. The drug requires a slow titration in order to achieve analgesia while avoiding dose-limiting side effects. This review examines the currently available information on this new analgesic. PMID- 17078785 TI - Recognizing and treating late-life generalized anxiety disorder: distinguishing features and psychosocial treatment. AB - Generalized anxiety disorder is a highly prevalent and under-recognized public health problem associated with significant impairment and serious disability. Generalized anxiety disorder is the most common anxiety disorder among adults over the age of 60 years. This article discusses distinguishing features of generalized anxiety disorder, such as disorder-specific worry domains and associated features. The relation of the disorder to normal aging, major depression and health is also discussed. Current psychosocial treatment strategies are outlined, with a particular focus on cognitive behavior therapy. PMID- 17078786 TI - Novel therapies for meningiomas. AB - Although advances in surgery, radiation therapy and stereotactic radiosurgery have significantly improved the treatment of meningiomas, there remains an important subset of patients who remain refractory to conventional therapy. Treatment with chemotherapeutic agents such as hydroxyurea and alpha-interferon has provided minimal benefit. In this review, the role of newly emerging novel therapies for meningiomas, with a focus on targeted molecular agents, will be discussed. PMID- 17078787 TI - Chemotherapy and targeted molecular therapies for brain metastases. AB - As therapy for systemic cancers improves, an increasing number of patients are developing brain metastases. Although conventional therapy with surgery, radiation therapy and radiosurgery has improved the outcome of a significant number of patients, many develop multiple lesions that are not amenable to standard treatments. In this review, the current role of chemotherapy and targeted molecular agents for brain metastases is summarized and future directions are discussed. PMID- 17078788 TI - Immunotherapy for patients with malignant glioma: from theoretical principles to clinical applications. AB - Malignant gliomas are the most common type of primary brain tumor and are in great need of novel therapeutic approaches. Advances in treatment have been very modest, significant improvement in survival has been lacking for many decades and prognosis remains dismal. Despite 'gross total' surgical resections and currently available radio-chemotherapy, malignant gliomas inevitably recur due to reservoirs of notoriously invasive tumor cells that infiltrate adjacent and nonadjacent areas of normal brain parenchyma. In principle, the immune system is uniquely qualified to recognize and target these infiltrative pockets of tumor cells, which have generally eluded conventional treatment approaches. In the span of the last 10 years, our understanding of the cancer-immune system relationship has increased exponentially, and yet, we are only beginning to tease apart the intricacies of the CNS and immune cell interactions. This article reviews the complex associations of the immune system with brain tumors. We provide an overview of currently available treatment options for malignant gliomas, existing gaps in our knowledge of brain tumor immunology, and molecular techniques and targets that might be exploited for improved patient stratification and design of 'custom immunotherapeutics'. We will also examine major new immunotherapy approaches that are being actively investigated to treat patients with malignant glioma, and identify some current and future research priorities in this area. PMID- 17078789 TI - Advances in strategies to improve drug delivery to brain tumors. AB - Brain tumors remain a significant health problem. Advances in the biology of the blood-brain barrier are improving the ability of researchers to target therapeutic peptides, small molecules and other drugs to brain tumors. Simple methods to improve blood-brain barrier penetration include chemical modification, glycosylation and pegylation. Drug-delivery vehicles, such as nanoparticles and liposomes, are also under study. Targeting vectors include natural ligands (e.g., epidermal growth factor) or monoclonal antibodies to receptors (e.g., transferrin or insulin). Other vector-mediated delivery approaches involve the conjugation of a therapeutic peptide or protein with a targeting molecule that can induce transcytosis across blood-brain barrier endothelial cells. The most commonly used vectors are peptidomimetic antibodies to endothelial receptors, such as the transferrin and insulin receptors. PMID- 17078790 TI - Response of gait deficits to neuromuscular electrical stimulation for stroke survivors. AB - Persistent gait deficits after stroke can cause falls, elevated energy cost and poor endurance. Coordination impairment is an underlying cause of gait deficits. Few efficacious interventions have been described that have targeted and measured restoration of coordinated gait components. Neuromuscular electrical stimulation can provide the critical gait practice characteristic of close-to-normal movements, by electrically inducing muscle contractions and coordinated movements that are not possible under volitional effort. Two-channel, surface neuromuscular electrical stimulation can be synchronized with phases of gait and can provide faster, more symmetrical neuromuscular electrical stimulation-assisted gait than gait with no neuromuscular electrical stimulation. Difficulties encountered during the use of surface neuromuscular electrical stimulation for gait training led to the development of neuromuscular electrical stimulation with implanted technologies. Implanted electrodes and/or stimulators proved to be feasible for gait training in stroke survivors. Gait training with a multichannel neuromuscular electrical stimulation system with implanted electrodes proved more advantageous than gait training without neuromuscular electrical stimulation, according to measures of volitional coordinated gait components (neuromuscular electrical stimulation deactivated). PMID- 17078791 TI - Atrial fibrillation and stroke prevention. AB - There is overwhelming evidence from randomized trials and systematic reviews to indicate the benefit of thromboprophylaxis in patients with atrial fibrillation. In moderate- to high-risk subjects, oral anticoagulation with warfarin reduces stroke by two-thirds, while aspirin reduces stroke by 22%. The latter result is similar to that seen for stroke reduction with antiplatelet therapy in vascular disease. Numerous studies have shown that less than half the patients eligible for warfarin therapy actually receive it and under- or overanticoagulation is common. This leads to many missed opportunities in optimizing stroke prevention in atrial fibrillation. The limitations of existing oral anticoagulants have resulted in the development of many new drugs. The aim of this review is to provide a brief overview of thromboprophylaxis in atrial fibrillation, and the opportunities for improvement in the provision made for thromboprophylaxis. PMID- 17078792 TI - Trigeminal-autonomic headaches in daily clinical practice. AB - Following the revised International Headache Society criteria, a group of short lasting headaches associated with autonomic symptoms, the so called trigeminal autonomic cephalgias, were newly recognized. The trigeminal autonomic cephalgias include cluster headache, paroxysmal hemicranias and a syndrome involving short lasting unilateral neuralform cephalgias with conjunctival injection and tearing (SUNCT) syndrome. In all of these syndromes, the half-sided head pain and cranial autonomic symptoms are prominent. All of the trigeminal autonomic cephalgias differ in duration, frequency and rhythmicity of the attacks, the intensity of pain and autonomic symptoms, as well as treatment options. This review gives a brief clinical description of the headache disorders and recent pathophysiological findings, as well as an overview of the treatment of cluster headache, paroxysmal hemicranias and SUNCT syndrome. PMID- 17078793 TI - Predicting treatment response in chronic, acquired demyelinating neuropathies. AB - Chronic inflammatory demyelinating polyradiculopathy is an immune-mediated neuropathy that was first described approximately 30 years ago. Since that time an increasingly wide spectrum of chronic acquired demyelinating polyneuropathies exhibiting different phenotypes, clinical course and treatment responses to immunomodulatory treatment have been described. Several new therapeutic agents have been prescribed for such conditions, some with promising results. This review summarizes what is presently known about the clinical courses, treatment responses and predictors of response of the chronic inflammatory demyelinating polyradiculopathy subgroups. PMID- 17078794 TI - Treatment of neuropsychiatric conditions associated with multiple sclerosis. AB - This article reviews the treatment of behavioral disturbances associated with multiple sclerosis. Pharmacological and psychotherapeutic treatment data, when available, are presented for five discrete conditions: major depression, bipolar affective disorder, anxiety, psychosis, pseudobulbar affect and cognitive dysfunction. Despite the paucity of empirical treatment data that characterizes all of these conditions, with the exception of pseudobulbar affect, the message from open-label trials and anecdotal experience is that therapy is often successful, leading to improvements in quality of life for patients. Thus, all health professionals involved in the care of multiple sclerosis patients should have a good working knowledge of the neurotherapeutics of multiple sclerosis related behavioral disorders. PMID- 17078795 TI - Guillain-Barre syndrome and its treatment. AB - Guillain-Barre syndrome typically presents with an acute ascending areflexic weakness, progressing over 4 weeks or less. The most common form of the disease is an acute inflammatory demyelinating polyneuropathy, but other forms with primarily axonal pathologies are well documented. The association of Guillain Barre syndrome with a range of antecedent infections, particularly Campylobacter jejuni enteritis, is also established. A range of serological and neurophysiological investigations can assist in making an accurate diagnosis. Background information about the syndrome and the evidence base for such treatments are discussed herein. PMID- 17078799 TI - Comparison of sow welfare in the Swedish deep-bedded system and the US crated-sow system. AB - The author estimates that the cost of swine production in the United States would increase by > $1 billion if the Swedish deep-bedded system were adopted as the only system in this country. The question is whether the added cost results in proportionately improved pig welfare. PMID- 17078800 TI - What is your diagnosis? Appendicular sarcoma. PMID- 17078801 TI - What is your diagnosis? Os clitoris or os penis with ambiguous genitalia. PMID- 17078802 TI - ECG of the Month. Second-degree AV block. PMID- 17078803 TI - Effect of certified health programs on the sale price of beef calves marketed through a livestock videotape auction service from 1995 through 2005. AB - OBJECTIVE: To quantify effects of certified health programs on the sale price of beef calves sold through a livestock videotape auction service. DESIGN: Longitudinal study. SAMPLE POPULATION: 26,502 lots representing 3,205,192 beef calves sold through a livestock videotape auction service between 1995 and 2005. PROCEDURES: Data describing each lot of beef calves that were marketed from 1995 through 2005 by a livestock videotape auction service were obtained from sale catalogues. For each year of the study, multiple regression analysis was used to quantify the effect of certified health programs on sale price. RESULTS: For each year of the study, beef calves that qualified for the 2 most intensive certified health programs sold for significantly higher prices, compared with prices for similar calves that were not in a certified health program, had not been vaccinated against respiratory tract viruses, and were not weaned before delivery. Price premiums for calves in the most intensive certified health program ranged from $2.47/100 lb (hundredweight [cwt]; 1 cwt equals 45.45 kg) in 1995 to $7.91/cwt in 2004. Price premiums paid for calves qualifying for the next most intensive certified health program ranged from $0.99/cwt in 1996 to $3.47/cwt in 2004. The percentage of the total number of lots in the 2 most intensive certified health programs increased over time. CONCLUSIONS AND CLINICAL RELEVANCE: Findings from this study indicated that implementation of the 2 most intensive certified health programs consistently increased the price of beef calves, and these price premiums increased over time. PMID- 17078804 TI - Extralabel use of penicillin in food animals. AB - Penicillin is one of the most commonly detected drug residues in tissues and milk, and is the antimicrobial for which information is most often sought through FARAD. PMID- 17078805 TI - The 2006 American Association of Feline Practitioners Feline Vaccine Advisory Panel report. AB - Vaccination is a medical procedure, and the decision to vaccinate should be based on a risk-based assessment for each cat and each vaccine. PMID- 17078806 TI - Detection of feline herpesvirus 1 DNA in skin biopsy specimens from cats with or without dermatitis. AB - OBJECTIVE: To compare detection rates of feline herpesvirus 1 (FHV-1) DNA in skin biopsy specimens from cats with herpetic dermatitis, cats with nonherpetic dermatitis, and cats without dermatitis. DESIGN: Prevalence survey. Animals-5 cats (9 biopsy specimens) with herpetic ulcerative dermatitis, 14 cats (17 biopsy specimens) with nonherpetic ulcerative dermatitis, and 8 cats (21 biopsy specimens) without clinically apparent skin lesions. PROCEDURES: A single-phase PCR assay was used to detect FHV-1 DNA in biopsy specimens. Assay results were compared with results of histologic examination. RESULTS: FHV-1 DNA was detected in all 9 biopsy specimens from the 5 cats with herpetic dermatitis and in 1 of 17 biopsy specimens from the 14 cats with nonherpetic dermatitis, but was not detected in any of the 21 biopsy specimens from the 8 cats without dermatitis. When results of histologic examination were used as the gold standard, sensitivity and specificity of the PCR assay were 100% and 95%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Results confirmed that FHV-1 DNA can be detected in the skin of cats with herpetic dermatitis and suggest that the virus may play a causative role in the disease. In addition, the PCR assay may be useful in confirming a diagnosis of herpetic dermatitis. PMID- 17078808 TI - Long-term survival and risk factors associated with biliary surgery in dogs: 34 cases (1994-2004). AB - OBJECTIVE: To determine factors associated with long-term survival after biliary surgery in dogs. DESIGN: Retrospective case series. ANIMALS: 34 dogs that underwent biliary surgery. PROCEDURES: Data extracted from medical records included sex, breed, body weight, age at surgery, history and clinical examination findings, preoperative and postoperative CBC, serum biochemical panel and coagulation profiles results, abdominal ultrasonographic findings, results of bacteriologic culture and histologic examination, surgical findings, postoperative complications, and survival time. Follow-up information was obtained from medical records or phone conversations with owners and referring veterinarians. RESULTS: Primary biliary findings included gallbladder mucocele (n = 20 dogs), inflammatory diseases (4), trauma (3), and neoplasia (1). Secondary biliary diseases included pancreatitis (n = 4), pancreatic neoplasia (1), and duodenal perforation (1). One- and 2-year survival rates were both 66%. Increasing age; gamma-glutamyltransferase activity; preanesthetic heart rate; BUN, phosphorus, and bilirubin concentrations; and the use of biliary diversion procedures were risk factors for death, although pancreatitis was not. However, poor long-term survival was associated with pancreatitis. CONCLUSIONS AND CLINICAL RELEVANCE: Long-term prognosis was guarded after biliary surgery in dogs. However, dogs that survived the early postoperative period had good long term prognosis. Dogs with pancreatitis had poor prognosis. Overall, the prognosis was worse for dogs that underwent a biliary diversion, compared with dogs that did not. PMID- 17078807 TI - Comparison of endoscopic and full-thickness biopsy specimens for diagnosis of inflammatory bowel disease and alimentary tract lymphoma in cats. AB - OBJECTIVE: To evaluate the accuracy of endoscopic biopsy (EB) specimens for diagnosis of alimentary tract lymphosarcoma in cats. DESIGN: Prospective study. ANIMALS: 22 cats with inflammatory bowel disease (IBD) or alimentary tract lymphosarcoma. PROCEDURES: Endoscopic biopsy specimens were obtained during endoscopy of the stomach and duodenum immediately prior to laparotomy or laparoscopic surgery, during which full-thickness biopsy (FTB) specimens were obtained. Accuracy of histopathologic diagnoses was compared between EB and FTB specimens. RESULTS: Lymphosarcoma was diagnosed in 10 cats on the basis of FTB specimens. Lymphosarcoma was detected in the jejunum and ileum in all 10 cats, in the duodenum in 9 cats, and in the stomach in 4 cats. In the same 10 cats, EB findings indicated a diagnosis of lymphosarcoma in 3 cats and were suggestive but inconclusive for lymphosarcoma in 3 cats. Lymphosarcoma was correctly diagnosed via gastric EB specimens in 3 of the 4 cats with gastric lymphosarcoma but evaluation of EB specimens led to an incorrect diagnosis of IBD in 4 cats with small intestinal lymphosarcoma. CONCLUSIONS AND CLINICAL RELEVANCE: EB specimens were useful for diagnosis of gastric lymphosarcoma but were not adequate for differentiating between IBD and lymphosarcoma in the small intestine. Because the most common sites of alimentary tract lymphosarcoma in cats are the jejunum and ileum, FTB specimens of those sites should be obtained via laparotomy or laparoscopy for accurate diagnosis. Laparoscopy may be a minimally invasive alternative to endoscopy and laparotomy for obtaining diagnostic biopsy specimens. PMID- 17078809 TI - Changes in central venous pressure and blood lactate concentration in response to acute blood loss in horses. AB - OBJECTIVE: To evaluate selected hemodynamic, blood gas, and biochemical responses to mild to moderate acute blood loss in standing, awake horses. DESIGN: Prospective study. ANIMALS: 7 healthy mares. PROCEDURES: Each horse was restrained in standing stocks, and its head was maintained in a neutral position; sedatives and tranquilizers were not administered. During a 1-hour period, blood was collected into collection bags by use of a suction pump. The rate of blood collection was approximately 16 mL/kg/h (7.3 mL/lb/h). Thirty minutes after blood collection, the blood was readministered at the same rate. Central venous pressure (CVP), central venous blood gas, blood lactate concentration, and heart rate were measured at baseline (after placement of catheters), after removal of blood, and after readministration of blood. RESULTS: In response to blood loss, CVP decreased and blood lactate concentration increased significantly, compared with baseline values; heart rate and results of central venous blood gas analysis did not change significantly. After readministration of blood, CVP returned to baseline value and blood lactate concentration approached baseline value. CONCLUSIONS AND CLINICAL RELEVANCE: Changes in CVP and blood lactate concentration appear to be early indicators of hypovolemia in horses, which may represent acute blood loss in trauma patients; these variables should be monitored to assess the potential need for blood transfusions. These variables can be used to monitor responses of horses to blood transfusions when whole blood is administered as the replacement fluid. PMID- 17078811 TI - Evaluation of ornithine carbamoyl transferase and other serum and liver-derived analytes in diagnosis of fatty liver and postsurgical outcome of left-displaced abomasum in dairy cows. AB - OBJECTIVE: To evaluate postsurgical outcome in dairy cows with left-displaced abomasum (LDA) with regard to severity of fatty liver and assess the usefulness of preoperative determination of serum ornithine carbamoyl transferase (OCT) activity, bile acids concentration, and other variables for evaluating liver function during the postsurgical convalescence period. DESIGN: Prospective study. ANIMALS: 68 Holstein cows. PROCEDURES: Blood and liver biopsy specimens were obtained during standing LDA surgery. Liver tissue was examined histologically and classified by severity of fatty change. Serum activities of liver-derived enzymes and concentrations of total lipids, triglycerides, bile acids, glucose, beta-hydroxybutyric acid, bilirubin, and nonesterified fatty acids were determined. RESULTS: Most cows with LDA and cows with severe fatty liver were detected within the first month after calving. Postsurgical outcome was related to severity of fatty liver. All cows that died had severe fatty liver. Serum activities of OCT, aspartate aminotransferase, and glutamate dehydrogenase and serum total bilirubin concentration were sensitive indicators of fatty liver. Serum bile acids concentration was not an accurate indicator of fatty liver. CONCLUSIONS AND CLINICAL RELEVANCE: Postsurgical outcome of cows undergoing surgery to correct LDA was related to fatty liver severity. Assessment of serum activities of OCT, aspartate aminotransferase, and glutamate dehydrogenase and serum total bilirubin concentration is recommended for diagnosis of fatty liver in dairy cows with LDA, whereas determination of bile acids concentration is not. The strong correlation between OCT activity and degree of hepatocellular damage supports use of this enzyme for assessing severity of fatty liver and predicting postsurgical outcome in cows with LDA. PMID- 17078812 TI - Diagnostic efficacy of a reverse transcriptase-polymerase chain reaction assay to screen cattle for persistent bovine viral diarrhea virus infection. AB - OBJECTIVE: To evaluate diagnostic efficacy of a reverse transcriptase-PCR assay performed on pooled supernatant from fresh tissue samples to screen cattle for persistent infection with bovine viral diarrhea virus (BVDV). DESIGN: Cross sectional study. SAMPLE POPULATION: 174 pooled supernatant samples representing 12,528 animals between 1 and 12 months old. PROCEDURES: The supernatant was collected from fresh tissue samples suspended in phosphate-buffered saline solution that had been submitted for testing for BVDV infection. Supernatant samples were pooled, with pool size limited to < or = 100 samples, and assayed with a reverse transcriptase-PCR assay for BVDV. Results of the PCR assay were compared with results of an antigen-capture ELISA performed on individual tissue samples. RESULTS: Results of the PCR assay were positive for 27 of the 174 pooled samples (mean pool size, 72 samples). For 23 of these 27 pooled samples, results of the ELISA were positive for 1 or more of the individual tissue samples represented in the pooled sample, whereas for 4 of these pooled samples, results of the ELISA were negative for all individual tissue samples represented in the pooled sample. Results of the ELISA were negative for all individual tissue samples represented in the 147 pooled samples with negative PCR assay results. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that the reverse transcriptase-PCR assay can be used to screen cattle for persistent BVDV infection, with calculated sensitivity of 100% (95% confidence interval, 85.2% to 100%) and calculated specificity of 97.5% (95% confidence interval, 93.4% to 99.3%). PMID- 17078814 TI - Prophages of Staphylococcus aureus Newman and their contribution to virulence. AB - Four prophages (phiNM1-4) were identified in the genome of Staphylococcus aureus Newman, a human clinical isolate. phiNM1, phiNM2 and phiNM4, members of the siphoviridae family, insert at different sites (poiA, downstream of isdB and geh) in the staphylococcal chromosome. phiNM3, a beta-haemolysin (hlb) converting phage, encodes modulators of innate immune responses (sea, sak, chp and scn) in addition to other virulence genes. Replication of phiNM1, phiNM2 and phiNM4 occurs in culture and during animal infection, whereas phiNM3 prophage replication was not observed. Prophages were excised from the chromosome and S. aureus variants lacking phiNM3 or phiNM1, phiNM2 and phiNM4 displayed organ specific virulence defects in a murine model of abscess formation. S. aureus Newman lacking all four prophages was unable to cause disease, thereby revealing essential contributions of prophages to the pathogenesis of staphylococcal infections. PMID- 17078815 TI - Acyl carrier protein/SpoT interaction, the switch linking SpoT-dependent stress response to fatty acid metabolism. AB - Bacteria respond to nutritional stresses by producing an intracellular alarmone, guanosine 5'-(tri)diphosphate, 3'-diphosphate [(p)ppGpp], which triggers the stringent response resulting in growth arrest and expression of resistance genes. In Escherichia coli, upon fatty acid or carbon starvation, SpoT enzyme activity switches from (p)ppGpp degradation to (p)ppGpp synthesis, but the signal and mechanism for this response remain totally unknown. Here, we characterize for the first time a physical interaction between SpoT and acyl carrier protein (ACP) using affinity co-purifications and two-hybrid in E. coli. ACP, as a central cofactor in fatty acid synthesis, may be an ideal candidate as a mediator signalling starvation to SpoT. Accordingly, we show that the ACP/SpoT interaction is specific of SpoT and ACP functions because ACP does not interact with the homologous RelA protein and because SpoT does not interact with a non-functional ACP. Using truncated SpoT fusion proteins, we demonstrate further that ACP binds the central TGS domain of SpoT, consistent with a role in regulation. The behaviours of SpoT point mutants that do not interact with ACP reveal modifications of the balance between the two opposite SpoT catalytic activities thereby changing (p)ppGpp levels. More importantly, these mutants fail to trigger (p)ppGpp accumulation in response to fatty acid synthesis inhibition, supporting the hypothesis that the ACP/SpoT interaction may be involved in SpoT-dependent stress response. This leads us to propose a model in which ACP carries information describing the status of cellular fatty acid metabolism, which in turn can trigger the conformational switch in SpoT leading to (p)ppGpp accumulation. PMID- 17078816 TI - Characterization of native and reconstituted exosome complexes from the hyperthermophilic archaeon Sulfolobus solfataricus. AB - The eukaryotic exosome is a protein complex with essential functions in processing and degradation of RNA. Exosome-like complexes were recently found in Archaea. Here we characterize the exosome of Sulfolobus solfataricus. Two exosome fractions can be discriminated by density gradient centrifugation. We show that the Cdc48 protein is associated with the exosome from the 30S-50S fraction but not with the exosome of the 11.3S fraction. While only some complexes contain Cdc48, the archaeal DnaG-like protein was found to be a core exosome subunit in addition to Rrp4, Rrp41, Rrp42 and Csl4. Assays with depleted extracts revealed that the exosome is responsible for major ribonucleolytic activity in S. solfataricus. Various complexes consisting of the Rrp41-Rrp42 hexameric ring and Rrp4, Csl4 and DnaG were reconstituted. Dependent on their composition, different complexes showed variations in RNase activity indicating functional interdependence of the subunits. The catalytic activity of these complexes and of the native exosome can be ascribed to the Rrp41-Rrp42 ring, which degrades RNA phosphorolytically. Rrp4 and Csl4 do not exhibit any hydrolytic RNase activity, either when assayed alone or in context of the complex, but influence the activity of the archaeal exosome. PMID- 17078813 TI - Cr(VI)-stimulated STAT3 tyrosine phosphorylation and nuclear translocation in human airway epithelial cells requires Lck. AB - Chronic inhalation of low amounts of Cr(VI) promotes pulmonary diseases and cancers through poorly defined mechanisms. SFKs (Src family kinases) in pulmonary airway cells may mediate Cr(VI) signalling for lung injury, although the downstream effectors of Cr(VI)-stimulated SFKs and how they relate to pathogenic gene induction are unknown. Therefore SFK-dependent activation of transcription factors by non-cytotoxic exposure of human bronchial epithelial cells to Cr(VI) was determined. Protein-DNA binding arrays demonstrated that exposing BEAS 2B cells to 5 microM Cr(VI) for 4 and 24 h resulted in increased protein binding to 25 and 43 cis-elements respectively, while binding to 12 and 16 cis-elements decreased. Of note, Cr(VI) increased protein binding to several STAT (signal transducer and activator of transcription) cis-elements. Cr(VI) stimulated acute tyrosine phosphorylation and nuclear translocation of STAT1 over a 4 h period and a prolonged activation of STAT3 that reached a peak between 48 and 72 h. This prolonged activation was observed for both STAT3alpha and STAT3beta. Immunofluorescent confocal microscopy confirmed that Cr(VI) increased nuclear localization of phosphorylated STAT3 for more than 72 h in both primary and BEAS 2B human airway cells. Cr(VI) induced transactivation of both a STAT3-driven luciferase reporter construct and the endogenous inflammatory gene IL-6 (interleukin-6). Inhibition with siRNA (small interfering RNA) targeting the SFK Lck, but not dominant-negative JAK (Janus kinase), prevented Cr(VI)-stimulated phosphorylation of both STAT3 isoforms and induction of IL-6. The results suggest that Cr(VI) activates epithelial cell Lck to signal for prolonged STAT3 activation and transactivation of IL-6, an important immunomodulator of lung disease progression. PMID- 17078817 TI - Truncated forms of IS911 transposase downregulate transposition. AB - IS911 naturally produces transposase (OrfAB) derivatives truncated at the C terminal end (OrfAB-CTF) and devoid of the catalytic domain. A majority species, OrfAB*, was produced at higher levels at 42 degrees C than at 30 degrees C suggesting that it is at least partly responsible for the innate reduction in IS911 transposition activity at higher temperatures. An engineered equivalent of similar length, OrfAB[1-149], inhibited transposition activity in vivo or in vitro when produced along with full-length transposase. We isolated several point mutants showing higher activity than the wild-type IS911 at 42 degrees C. These fall into two regions of the transposase. One, located in the N-terminal segment of OrfAB, lies between or within two regions involved in protein multimerization. The other is located within the C-terminal catalytic domain. The N-terminal mutations resulted in reduced levels of OrfAB* while the C-terminal mutation alone appeared not to affect OrfAB* levels. Combination of N- and C-terminal mutations greatly reduced OrfAB* levels and transposition was concomitantly high even at 42 degrees C. The mechanism by which truncated transposase species are generated and how they intervene to reduce transposition activity is discussed. While transposition activity of these multiply mutated derivatives in vivo was resistant to temperature, the purified OrfAB derivatives retained an inherent temperature-sensitive phenotype in vitro. This clearly demonstrates that temperature sensitivity of IS911 transposition is a complex phenomenon with several mechanistic components. These results have important implications for the several other transposons and insertion sequences whose transposition has also been shown to be temperature-sensitive. PMID- 17078821 TI - Evidence of nursing shortages or a shortage of evidence? PMID- 17078820 TI - What's the scoop on the nursing shortage? PMID- 17078818 TI - RyhB small RNA modulates the free intracellular iron pool and is essential for normal growth during iron limitation in Escherichia coli. AB - The small RNA RyhB has recently been shown to negatively regulate a number of mRNAs encoding dispensable iron-using proteins in Escherichia coli. The resulting decrease in the synthesis of iron-using proteins is thought to spare iron in order to ensure its availability for iron-requiring proteins that are indispensable. Indeed, the expression of RyhB from a heterologous promoter activates the iron-sensing repressor Fur, which suggests an increase in the pool of free intracellular iron (iron-sparing). In accordance with these observations, we report here that RyhB expression increases the concentration of free intracellular iron, as shown by direct measurements of the metal in whole cells by electron paramagnetic resonance spectroscopy. Our data also suggest that iron sparing originates from rapid uptake of extracellular iron and not from already internalized metal. Furthermore, RyhB is shown to be essential for normal bacterial growth and survival during iron starvation, which is consistent with previous data describing the function of the small RNA. Overall, our data demonstrate that, by regulating synthesis of nonessential iron-using proteins, the small RNA RyhB ensures that the iron is directed towards the iron-requiring enzymes that are indispensable. PMID- 17078822 TI - Theoretical model of coping among relatives of patients in intensive care units: a simultaneous concept analysis. AB - This paper reports the development of a theoretical model of relatives' coping approaches during the patient's intensive care unit stay and subsequent recovery at home by performing an analysis of concepts generated from two empirically grounded, theoretical studies in this area. BACKGROUND: When supporting relatives of intensive care unit patients, it is important that nurses have access to evidence-based knowledge of relatives' coping approaches during the period of illness and recovery. METHOD: Simultaneous concept analysis was used to refine and combine multiple coping concepts into a theoretical model of coping. The concepts were generated in two previous empirical studies of relatives' coping approaches during mechanically ventilated patients' intensive care unit stays and recovery at home. FINDINGS: The theoretical model was developed in 2004-2005 and illustrates the effectiveness of different coping approaches in relation to each other and to social support. Definitions summarizing each coping approach and containing the knowledge gained through the simultaneous concept analysis method were also formulated. CONCLUSION: This middle-range theory of relatives' coping approaches may make a valuable contribution to international intensive care unit nursing practice, especially as it is based on empirical studies and may therefore serve as a basis for the development of future clinical guidelines. However, the theoretical model needs to be empirically validated before it can be used. PMID- 17078823 TI - Development of a framework for person-centred nursing. AB - This paper presents the development and content of a person-centred nursing framework. BACKGROUND AND RATIONALE: Person-centred is a widely used concept in nursing and health care generally, and a range of literature articulates key components of person-centred nursing. This evidence base highlights the links between this approach and previous work on therapeutic caring. METHODS: The framework was developed through an iterative process and involved a series of systematic steps to combine two existing conceptual frameworks derived from empirical studies. The process included the mapping of original conceptual frameworks against the person-centred nursing and caring literature, critical dialogue to develop a combined framework, and focus groups with practitioners and co-researchers in a larger person-centred nursing development and research project to test its face validity. FINDINGS: The person-centred nursing framework comprises four constructs -prerequisites, which focus on the attributes of the nurse; the care environment, which focuses on the context in which care is delivered; person-centred processes, which focus on delivering care through a range of activities; and expected outcomes, which are the results of effective person-centred nursing. The relationship between the constructs suggests that, to deliver person-centred outcomes, account must be taken of the prerequisites and the care environment that are necessary for providing effective care through the care processes. CONCLUSION: The framework described here has been tested in a development and research project in an acute hospital setting. Whilst there is an increasing empirical base for person-centred nursing, as yet little research has been undertaken to determine its outcomes for patients and nurses. The framework developed can be described as a mid-range theory. Further testing of the framework through empirical research is required to establish its utility for nursing practice and research. PMID- 17078824 TI - Development and testing of a Clinical Learning Environment Diagnostic Inventory for baccalaureate nursing students. AB - Aim. This paper reports the development and testing of the psychometric properties of the Clinical Learning Environment Diagnostic Inventory for baccalaureate nursing students. Background. The quality of the clinical learning environment is an essential factor in determining the quality of nursing students' clinical experience. It is also well-recognized that the clinical setting can be a source of stress and anxiety for students. To design an optimal clinical learning environment for students, instruments based on the cognitive and socio-emotional aspects of the learning process are necessary. Method. The hypothetical construct of the clinical learning environment was derived from a comprehensive review of the literature, including experiential learning theory and the epistemology of practice. Developing the instrument involved the collection of items through semi-structured interviews, assessing content validity and determining the scaling. After a pilot study, the instrument was tested with 312 undergraduate students and 157 preceptors between September and December 2004. The factor structure was determined by exploratory factor analysis. Results. Exploratory factor analysis produced a five-factor solution that was similar to the hypothetical model. Cronbach's alpha internal consistency reliability coefficients ranged from 0.65 to 0.77 across the five factors. Three week test-retest reliability coefficients ranged from 0.59 to 0.74. Criterion related validity and construct validity were estimated by the correlation between the Clinical Learning Environment Diagnostic Inventory and appropriate other instruments and a set of comparison data obtained from the students and preceptors. Conclusion. The findings yielded acceptable estimates of reliability and validity of the Clinical Learning Environment Diagnostic Inventory. Therefore, this instrument is considered useful for assessing learning environments in clinical settings. PMID- 17078825 TI - 'Best practice' in focus group research: making sense of different views. AB - AIM: The aim of this paper is to identify the broad epistemological debates which underpin conflicting statements on 'rigour' and 'good practice' in qualitative research; to relate divergences in statements of 'good practice' in focus group design made by the pre-eminent commentators on focus group methodology to these broader epistemological debates; and to stimulate further reflection on the range of possible uses for focus groups in health services research. Considerations of the analysis of focus group data are beyond the scope of this paper. DISCUSSION: Focus groups are a popular form of qualitative data collection, and may be defined as a particular form of group interview intended to exploit group dynamics. While qualitative research may be broadly characterized as concerned with exploring people's lived experiences and perspectives in context, it is a heterogeneous field incorporating many theoretical traditions. Consequently, qualitative researchers may be informed by a wide range of assumptions about the nature of knowledge (epistemology). These assumptions, whether implicit or explicit, have important consequences for claims about rigour and 'good practice' in data collection. Thus, while there is broad agreement over the general form of focus groups, statements of 'good practice' in terms of its application are varied. A close reading of texts by the two pre-eminent commentators on the practical application of focus groups identifies differences in 'best practice' focus group design related to their respective epistemological assumptions, and differences principally related to sampling techniques, composition of groups, the perceived role of group interaction and the nature of inference. CONCLUSION: Explicit consideration of the epistemological basis of divergent statements of 'best practice' in focus group design forces health services researchers to balance the demands of theory with the practicalities of conducting focus group research within complex host organisations; and encourages readers to apply appraisal criteria appropriate to the stated intentions of researchers. PMID- 17078826 TI - Mapping the organizational culture research in nursing: a literature review. AB - AIM: This paper reports a critical review of nursing organizational culture research studies with the objectives of: (1) reviewing theoretical and methodological characteristics of the studies and (2) drawing inferences specific to the state of knowledge in this field. BACKGROUND: Organizational culture is regarded as significant in influencing research use in clinical practice yet it is not understood how culture shapes practitioners' behaviours. Only one review of this empirical literature in nursing has been completed. METHOD: Using selected computerized databases, published nursing research studies in English that examine organizational culture were accessed. Organizational culture studies were categorized using Hatch's three perspectives on organizational culture: (1) modern, (2) symbolic-interpretive and (3) postmodern. The review was conducted in 2005. RESULTS: Twenty-nine studies were in the final data set. Results pointed to variations in cultural definitions and incorporation of organizational sciences theory. In classifying the studies, modern perspectives dominated (n = 22), symbolic-interpretive approaches were an emerging group (n = 6) and one study was unclassifiable. Our results expand current cultural instrument reviews by pinpointing tools that have been previously overlooked and by identifying ongoing theoretical and methodological challenges for researchers. CONCLUSION: An exclusive reliance on modernistic approaches in organizational culture research cannot yield a complete understanding of the phenomenon. Rather, the field could benefit from a variety of cultural approaches. In a similar vein, researchers need to be mindful of the terminology and the unit of analysis they use in their research, as these are the two largest research challenges. PMID- 17078827 TI - Nursing diagnoses, interventions and outcomes - application and impact on nursing practice: systematic review. AB - AIM: This paper reports a systematic review on the outcomes of nursing diagnostics. Specifically, it examines effects on documentation of assessment quality; frequency, accuracy and completeness of nursing diagnoses; and on coherence between nursing diagnoses, interventions and outcomes. BACKGROUND: Escalating healthcare costs demand the measurement of nursing's contribution to care. Use of standardized terminologies facilitates this measurement. Although several studies have evaluated nursing diagnosis documentation and their relationship with interventions and outcomes, a systematic review has not been carried out. METHOD: A Medline, CINAHL, and Cochrane Database search (1982-2004) was conducted and enhanced by the addition of primary source and conference proceeding articles. Inclusion criteria were established and applied. Thirty-six articles were selected and subjected to thematic content analysis; each study was then assessed, and a level of evidence and grades of recommendations assigned. FINDINGS: Nursing diagnosis use improved the quality of documented patient assessments (n = 14 studies), identification of commonly occurring diagnoses within similar settings (n = 10), and coherence among nursing diagnoses, interventions, and outcomes (n = 8). Four studies employed a continuing education intervention and found statistically significant improvements in the documentation of diagnoses, interventions and outcomes. However, limitations in diagnostic accuracy, reporting of signs/symptoms, and aetiology were also reported (14 studies). One meta-analysis of eight trials including 1497 patients showed no evidence that standardized electronic documentation of nursing diagnosis and related interventions led to better nursing outcomes. CONCLUSION: Despite variable results, the trend indicated that nursing diagnostics improved assessment documentation, the quality of interventions reported, and outcomes attained. The study reveals deficits in reporting of signs/symptoms and aetiology. Consequently, staff educational measures to enhance diagnostic accuracy are recommended. The relationships among diagnoses, interventions and outcomes require further evaluation. Studies are needed to determine the relationship between the quality of documentation and practice. PMID- 17078828 TI - Determinants of tenure in allied health and nursing education. AB - AIM: This paper presents a prospective cross-sectional study that investigated the tenure rate and the primary criterion use in granting tenure in nursing and allied health education in the United States of America. BACKGROUND: Given the recent trend by highly skilled professionals to seek employment in other countries, a clear understanding of the conditions of service in higher education is important to educators contemplating the relocating to another country. The preponderance of the published literature on academic tenure is from the United States of America, where educators continue to debate the value of the tenure system and the criteria to be used in tenure decisions. METHOD: We surveyed the deans of National League for Nursing accredited programmes (n = 187) and deans of allied health programmes belonging to the association of schools of allied health professions (n = 75) in the United States of America. The questionnaire sought demographic and institution-related information, tenure rate and weightings attached to teaching, scholarship and service in tenure decision. The data were collected in 2002. FINDINGS: Allied health and nursing educators had 47% and 35% tenure rates, respectively. The overwhelming majority of the deans in our study, 77%, ranked teaching as the primary criterion used in tenure decisions in their institutions. On the other hand, fewer than 25% rated scholarship and fewer than 5% rated service as the most important criterion used for tenure in their institution. The responses of the deans were modulated by the type and ownership of the institution in which they were employed and the characteristics of the educators. CONCLUSION: The implications for preparing future educators in the United States of America for long-term careers in allied health and nursing professions are that: (1) teaching is less highly valued in research-oriented universities; and (2) heavy teaching workloads may be detrimental to the chances of obtaining tenure. Replication of the study in other countries would have the potential to facilitate the employment mobility and educator exchange. PMID- 17078829 TI - Low-perceived work ability, ageing and intention to leave nursing: a comparison among 10 European countries. AB - AIM: This paper reports a study exploring nurses' perceived work ability and its associations with age and intention to leave nursing in a representative sample of registered nurses in 10 european countries. BACKGROUND: Throughout Europe, there is now a substantial shortage of Registered Nurses and unless steps are taken to reverse this trend, numbers are likely to decline further. A study exploring nurses' perceived work ability will provide baseline evidence, which may lead to improved working conditions and increased nursing retention. METHODS: A cross-sectional study design was employed. Questionnaire data were collected from 25,976 nurses in 10 member states of the European Union between October 2002 and June 2003. The response rate was 52.9% for the total investigation and varied between countries from 32.4% to 76.9%. Perceived work ability was assessed using the Work Ability Index. Intention to leave was measured by asking nurses how often they thought about leaving nursing. Data were examined using analysis of covariance and adjusted logistic regression. RESULTS: In all 10 European countries, scores on the Work Ability Index were significantly lower (P < 0.01) among older nurses (>or=45 years). Work ability varied among countries and differences between younger and older nurses were more pronounced in some countries. In all countries, there was a significant association between low Work Ability Index and intention to leave nursing (odds ratios between 1.98 and 21.46), especially among younger nurses. The association between work ability and intention to leave was most marked for those items on the Work Ability Index which explored subjective rather than objective aspects of work ability. CONCLUSION: Attempts to redress nursing shortages could include institutional policies to sustain work ability through better working conditions, improving quality of the working environment and finding suitable alternative nursing work for those no longer able to cope in their current post. These approaches should include nurses in all age categories. PMID- 17078830 TI - The introduction of innovative nursing and midwifery roles: the perspective of healthcare managers. AB - AIM: This paper reports a study exploring issues arising from the introduction of innovative nursing and midwifery roles from the perspective of healthcare managers. BACKGROUND: In recent years, the number of innovative roles in nursing and midwifery has increased dramatically across all areas of health care. A comprehensive literature review has shown that there is a scarcity of empirical studies to inform policy or the organizational context of this rapidly changing situation. METHOD: Semi-structured interviews were conducted with all Directors of Nursing, Chief Nurses and Directors of Primary Care in the Health and Social Services Trusts and Boards in Northern Ireland. This study formed the first phase of a larger exploration of innovative roles. Data were collected from July to October 2004. FINDINGS: The findings confirm that in recent years there has been a considerable increase in the number and type of innovative nursing and midwifery roles. Themes that emerged from the data included professional identity and number of innovative roles; stimuli for role development; service commissioning; infrastructure support; and perceived value for money. CONCLUSION: Analysis of the impact of innovative nursing and midwifery roles on other members of staff, notably healthcare assistants, is needed. The internationally increasing number of innovative roles need to be evaluated further with regard to their effect on care outcomes for patients, their families and communities, their cost-effectiveness and to how best to secure the support and commitment of colleagues. Further study should also be undertaken to determine whether innovative role-holders are pioneers in the development of future nursing, or whether they are responding to the workforce needs of the medical profession or to the requirements of the health service to cut costs. PMID- 17078831 TI - Long working hours increase the risk of sharp and needlestick injury in nurses: the need for new policy implication. AB - AIM: This paper reports a study to determine the sharp and needlestick injury incidence in nurses working at a university hospital and the contributing factors. BACKGROUND: Although it is generally felt that working in the healthcare sector is clean and without risk, healthcare staff and especially physicians and nurses who generally work very long hours are actually exposed to various occupational risks. Sharps and needlestick injuries are important problems for healthcare workers as they increase the risk of spread of infection. METHOD: A self-administered questionnaire was completed in October 2005 by 449 of the 516 nurses working at a Turkish hospital (response rate 87.0%). RESULTS: The percentage of nurses experiencing a sharp or needlestick injury during their professional life was 79.7%. The incidence of exposure to sharp or needlestick injury in the last year was 68.4%. The factors increasing the rate of sharp and needlestick injury were: age 24 years and less, 7.5). When the mean total annual costs was adjusted by the mean % of patients on IT in our Unit (31%) the result was 19589 euros. The key drivers for direct costs were IT in low EDSS stages, and caregiver costs in high stages. Indirect costs were assessed in terms of the loss of productivity when patients stop working. Direct costs accounted for around 60% of total costs in all EDSS groups. IT accounts from 78% to 11% of direct costs, and decreased as disability progressed. CONCLUSION: The total mean social costs of MS in a cohort from Baix Llobregat (Catalonia) were estimated at 24,272 euros per patient/year, and ranged between 14,327 euros (EDSS = 0) and 52,841 euros (EDSS = 7.5-9.5). Total costs, and particularly informal and direct costs, increased as the disability progressed. IT should be able to delay the progression of disability to be efficient and not only effective. PMID- 17078880 TI - A cross-sectional study of the relation between symptoms and physical findings in computer operators. AB - BACKGROUND: The character of upper limb disorder in computer operators is subject to debate. A peripheral nerve-involvement is suggested from the common presence of a triad of symptoms consisting of pain, paraestesiae and subjective weakness, and from physical findings suggesting neuropathy. This study aimed to examine the outcome of a detailed neurological examination in computer operators and to compare findings with the presence of symptoms. METHODS: 96 graphical computer operators answered a modified Nordic Questionnaire including information on perceived pain in the shoulder, elbow, and wrist/hand scored for each region on a VAS-scale 0-9. In addition, they underwent a physical examination including the subjective assessment of the individual function of 11 upper limb muscles, of algesia in five and vibratory threshold in three territories, respectively, and of mechanosensitivity of nerves at seven locations. In order to reflect an involvement of the brachial plexus (chord level), the posterior interosseous nerve and the median nerve at elbow level we defined three patterns of neurological findings illustrating the course of nerves and their innervation. The pain scores summarized for the three upper limb regions (min. = 0, max = 27) in the mouse-operating and contralateral limbs were compared by a Wilcoxon test and the relation to each physical item analyzed by Kendall's rank correlation. The relation of summarized pain to each pattern was studied by application of a test of the trend across ordered groups (patterns). RESULTS: Pain, paraestesiae and subjective weakness was reported for 67, 23, and 7 mouse-operating limbs, respectively, with the summarized pain scores exceeding 4 in 33 limbs. Abnormal physical findings were prevalent. The summarized pain was significantly related to a reduced function in five muscles, to mechanical allodynia at one location and to elevated threshold to vibration in two territories. Brachial plexopathy was diagnosed in 9/2, median neuropathy in 13/5 and posterior interosseous neuropathy in 13/8 mouse operating/contralateral limbs, respectively. The summarized pain was significantly higher in the mouse-operating limbs and in limbs with any of the defined patterns. There was a significant trend between the summarized pain and the summarized scores for the items contained in each pattern. CONCLUSION: This small-scale study of a group of computer-operators currently in work and with no or minor upper limb symptoms has indicated in symptomatic subjects the presence of peripheral nerve-afflictions with specific locations. PMID- 17078881 TI - Adaptive evolution of Hox-gene homeodomains after cluster duplications. AB - BACKGROUND: Hox genes code for homeodomain-containing transcription factors that function in cell fate determination and embryonic development. Hox genes are arranged in clusters with up to 14 genes. This archetypical chordate cluster has duplicated several times in vertebrates, once at the origin of vertebrates and once at the origin of gnathostoms, an additional duplication event is associated with the origin of teleosts and the agnanths, suggesting that duplicated Hox cluster genes are involved in the genetic mechanisms behind the diversification of vertebrate body plans, and the origin of morphological novelties. Preservation of duplicate genes is promoted by functional divergence of paralogs, either by subfunction partitioning among paralogs or the acquisition of a novel function by one paralog. But for Hox genes the mechanisms of paralog divergence is unknown, leaving open the role of Hox gene duplication in morphological evolution. RESULTS: Here, we use several complementary methods, including branch-specific dN/dS ratio tests, branch-site dN/dS ratio tests, clade level amino acid conservation/variation patterns, and relative rate ratio tests, to show that the homeodomain of Hox genes was under positive Darwinian selection after cluster duplications. CONCLUSION: Our results suggest that positive selection acted on the homeodomain immediately after Hox clusters duplications. The location of sites under positive selection in the homeodomain suggests that they are involved in protein-protein interactions. These results further suggest that adaptive evolution actively contributed to Hox-gene homeodomain functions. PMID- 17078882 TI - Reproducibility of target volumes generated using uncoached 4-dimensional CT scans for peripheral lung cancer. AB - BACKGROUND: 4-dimensional CT (4DCT) scans are increasingly used to account for mobility during radiotherapy planning. As variations in respiratory patterns can alter observed motion, with consequent changes in the generated target volumes, we evaluated the reproducibility of 4D target volumes generated during repeat uncoached quiet respiration. METHODS: A retrospective analysis was performed on two successive scans (4DCT1 and 4DCT2) generated at the same scanning session for 26 patients with peripheral lung cancer treated with stereotactic radiotherapy (SRT). The volume and position of planning target volumes (PTV4DCT1 and PTV4DCT2) contoured on both scans were compared, and a dosimetric analysis performed. A SRT plan optimized for each PTV was sequentially applied to the other PTV, and coverage by the 80% isodose was evaluated. Color intensity projections (CIP) were used to evaluate regions of underdosage. RESULTS: No significant volumetric differences were observed between the two PTVs (t-Test p = 0.60). The average displacement of the center of mass between corresponding PTVs was 1.4 +/- 1.0 mm, but differences in position were 2.0 mm or greater in 5 cases (19%). Coverage of both PTVs by the 80% prescription isodose exceeded 90% for all but one patient. For the latter, the prescription isodose covered only 82.5% of PTV4DCT1. CIP analysis revealed that the region of underdosage was an end-inspiratory position occupied by the tumor for only 10-20% of the respiratory cycle. CONCLUSION: In nearly all patients with stage I lung cancer, the PTV derived from a single uncoached 4DCT achieves dosimetric coverage that is similar to that achieved using two such consecutive scans. PMID- 17078883 TI - Costs of shoulder pain in primary care consulters: a prospective cohort study in The Netherlands. AB - BACKGROUND: Shoulder pain is common in primary care, and has an unfavourable outcome in many patients. Information on the costs associated with health care use and loss of productivity in patients with shoulder pain is very scarce. The objective of this study was to determine shoulder pain related costs during the 6 months after first consultation in general practice METHODS: A prospective cohort study consisting of 587 patients with a new episode of shoulder pain was conducted with a follow-up period of 6 months. Data on costs were collected by means of a cost diary during 6 months. RESULTS: 84% of the patients completed all cost diaries. The mean consumption of direct health care and non-health related care was low. During 6 months after first consultation for shoulder pain, the mean total costs a patient generated were 689 euro. Almost 50% of this total concerned indirect costs, caused by sick leave from paid work. A small proportion (12%) of the population generated 74% of the total costs. CONCLUSION: The total costs in the 6 months after first consultation for shoulder pain in primary care, mostly generated by a small part of the population, are not alarmingly high. PMID- 17078884 TI - Obstructive sleep apnoea is independently associated with the metabolic syndrome but not insulin resistance state. AB - Obstructive sleep apnoea (OSA) is a cardio-metabolic disorder. Whether metabolic syndrome (MS), insulin resistance (IR) and albuminuria are independently associated with OSA is unclear, but defining the interactions between OSA and various cardiovascular (CV) risk factors independent of obesity facilitates the development of therapeutic strategies to mitigate their increased CV risks. We prospectively recruited 38 subjects with OSA and 41 controls. Anthropometric measurements, glucose, lipids, insulin and blood pressure (BP) were measured after an overnight fast. IR state was defined as homeostasis model assessment (HOMA) value >3.99 and MS diagnosed according to the International Diabetes Federation (IDF) criteria. Subjects with OSA were more obese, more insulin resistant, more hyperglycaemic, had higher Epworth score (measure of day time somnolence) and systolic blood pressure levels. The prevalence of MS was higher in OSA compared with non-OSA subjects (74% vs 24%, p < 0.001). The prevalence of microalbuminuria in both groups was negligible. Logistic regression adjusted for age, BMI and smoking showed that the patient with OSA was 5.9 (95% CI 2.0-17.6) times more likely to have MS than non-OSA patient. Triglyceride (p = 0.031), glucose (0.023) and Epworth score (0.003) values were independently associated with OSA after adjusting for BMI and other covariates whilst IR status was found not to be significant. Using the ROC curve analysis, we found that a waist circumference of >103 cm would predict MS in patients with OSA at 75-78% sensitivity and 61-64% specificity. The agreement between MS and IR state in this cohort is poor. Thus, OSA is associated with MS independent of obesity predominantly due to increased triglyceride, glucose and Epworth score values but not IR or microalbuminuria status. This observation suggests an alternative pathogenic factor mediating the increased cardiovascular risk in patients with OSA and MS, other than that due to IR. The independent link between Epworth score and MS in patients with OSA implicates the role of daytime sleepiness and chronic hypoxia as a potential mediator. Given the discordant between MS and IR state, measurement of waist is useful for predicting mainly MS but not insulin resistance status in patients with OSA. Appropriate pharmacological intervention targeting these independent factors is important in reducing the increased CV risks among patients with OSA. PMID- 17078885 TI - Defective ALK5 signaling in the neural crest leads to increased postmigratory neural crest cell apoptosis and severe outflow tract defects. AB - BACKGROUND: Congenital cardiovascular diseases are the most common form of birth defects in humans. A substantial portion of these defects has been associated with inappropriate induction, migration, differentiation and patterning of pluripotent cardiac neural crest stem cells. While TGF-beta-superfamily signaling has been strongly implicated in neural crest cell development, the detailed molecular signaling mechanisms in vivo are still poorly understood. RESULTS: We deleted the TGF-beta type I receptor Alk5 specifically in the mouse neural crest cell lineage. Failure in signaling via ALK5 leads to severe cardiovascular and pharyngeal defects, including inappropriate remodeling of pharyngeal arch arteries, abnormal aortic sac development, failure in pharyngeal organ migration and persistent truncus arteriosus. While ALK5 is not required for neural crest cell migration, our results demonstrate that it plays an important role in the survival of post-migratory cardiac neural crest cells. CONCLUSION: Our results demonstrate that ALK5-mediated signaling in neural crest cells plays an essential cell-autonomous role in the pharyngeal and cardiac outflow tract development. PMID- 17078886 TI - Mining expressed sequence tags identifies cancer markers of clinical interest. AB - BACKGROUND: Gene expression data are a rich source of information about the transcriptional dis-regulation of genes in cancer. Genes that display differential regulation in cancer are a subtype of cancer biomarkers. RESULTS: We present an approach to mine expressed sequence tags to discover cancer biomarkers. A false discovery rate analysis suggests that the approach generates less than 22% false discoveries when applied to combined human and mouse whole genome screens. With this approach, we identify the 200 genes most consistently differentially expressed in cancer (called HM200) and proceed to characterize these genes. When used for prediction in a variety of cancer classification tasks (in 24 independent cancer microarray datasets, 59 classifications total), we show that HM200 and the shorter gene list HM100 are very competitive cancer biomarker sets. Indeed, when compared to 13 published cancer marker gene lists, HM200 achieves the best or second best classification performance in 79% of the classifications considered. CONCLUSION: These results indicate the existence of at least one general cancer marker set whose predictive value spans several tumor types and classification types. Our comparison with other marker gene lists shows that HM200 markers are mostly novel cancer markers. We also identify the previously published Pomeroy-400 list as another general cancer marker set. Strikingly, Pomeroy-400 has 27 genes in common with HM200. Our data suggest that a core set of genes are responsive to the deregulation of pathways involved in tumorigenesis in a variety of tumor types and that these genes could serve as transcriptional cancer markers in applications of clinical interest. Finally, our study suggests new strategies to select and evaluate cancer biomarkers in microarray studies. PMID- 17078888 TI - Suture midface suspension. AB - OBJECTIVE: To describe a simple and effective facelift technique useful as an adjunct to other oculoplastic procedures METHODS: Retrospective, non-comparative case series. Thirty five patients undergoing suture midface suspension from 1998 to 2000. Suspension sutures were passed from the nasolabial fold to the temporalis fascia to elevate the midface and the corner of the mouth. RESULTS: A satisfactory and stable outcome is obtained in 2 years of follow up. CONCLUSION: Suture midface suspension is a safe and effective technique for the management of midface descent. PMID- 17078887 TI - Multi-locus phylogeny of dolphins in the subfamily Lissodelphininae: character synergy improves phylogenetic resolution. AB - BACKGROUND: Dolphins of the genus Lagenorhynchus are anti-tropically distributed in temperate to cool waters. Phylogenetic analyses of cytochrome b sequences have suggested that the genus is polyphyletic; however, many relationships were poorly resolved. In this study, we present a combined-analysis phylogenetic hypothesis for Lagenorhynchus and members of the subfamily Lissodelphininae, which is derived from two nuclear and two mitochondrial data sets and the addition of 34 individuals representing 9 species. In addition, we characterize with parsimony and Bayesian analyses the phylogenetic utility and interaction of characters with statistical measures, including the utility of highly consistent (non homoplasious) characters as a conservative measure of phylogenetic robustness. We also explore the effects of removing sources of character conflict on phylogenetic resolution. RESULTS: Overall, our study provides strong support for the monophyly of the subfamily Lissodelphininae and the polyphyly of the genus Lagenorhynchus. In addition, the simultaneous parsimony analysis resolved and/or improved resolution for 12 nodes including: (1) L. albirostris, L. acutus; (2) L. obscurus and L. obliquidens; and (3) L. cruciger and L. australis. In addition, the Bayesian analysis supported the monophyly of the Cephalorhynchus, and resolved ambiguities regarding the relationship of L. australis/L. cruciger to other members of the genus Lagenorhynchus. The frequency of highly consistent characters varied among data partitions, but the rate of evolution was consistent within data partitions. Although the control region was the greatest source of character conflict, removal of this data partition impeded phylogenetic resolution. CONCLUSION: The simultaneous analysis approach produced a more robust phylogenetic hypothesis for Lagenorhynchus than previous studies, thus supporting a phylogenetic approach employing multiple data partitions that vary in overall rate of evolution. Even in cases where there was apparent conflict among characters, our data suggest a synergistic interaction in the simultaneous analysis, and speak against a priori exclusion of data because of potential conflicts, primarily because phylogenetic results can be less robust. For example, the removal of the control region, the putative source of character conflict, produced spurious results with inconsistencies among and within topologies from parsimony and Bayesian analyses. PMID- 17078889 TI - Mesothelioma with non-pleural malignancy: a red herring or just an uncommon pairing? AB - Malignant pleural mesothelioma (MPM) is a highly aggressive cancer of the pleura with a well-established male predominance and causative link with asbestos exposure. We report four cases of female patients with MPM referred for palliation of symptoms thought to be due to previous non-pleural malignancy.With emerging novel treatments for MPM, this article discusses four unusual cases of MPM occurring in the setting of other malignancy, highlights the importance of considering a primary diagnosis of MPM even in patients with other malignancy, and reinforces the benefits of video-assisted surgical biopsy which allows simultaneous diagnosis and treatment. PMID- 17078890 TI - Development of a GIS-based, real-time Internet mapping tool for rabies surveillance. AB - BACKGROUND: Oral rabies vaccination programs have been implemented to control the spread of wildlife rabies in the United States. However, current surveillance systems are inadequate for the efficient management and evaluation of these large scale vaccine baiting programs. With this in mind, a GIS-based rabies surveillance database and Internet mapping application was created. This surveillance system, RabID, provides a new resource for the rapid mapping and dissemination of data on animal rabies cases in relation to unaffected, enzootic, and baited areas where current interventions are underway. RESULTS: RabID is a centralized database for diagnostic and demographic information collected by local, state, and federal agencies involved in rabies surveillance. The geo referenced database remits data to an Internet-accessible mapping application that displays rabies surveillance data in relation to environmental and geographic features. CONCLUSION: RabID provides a pioneering example of the power of geographically based Internet-accessible, infectious disease surveillance. This surveillance system was developed from existing technology and is readily adaptable to other infectious diseases and may be particularly useful for zoonoses. The development and application of public health informatics technology may enhance the effectiveness of public health interventions and allow better evaluation of public health interventions. PMID- 17078891 TI - HIV-1 infection and CD4 T cell depletion in the humanized Rag2-/-gamma c-/- (RAG hu) mouse model. AB - BACKGROUND: The currently well-established humanized mouse models, namely the hu PBL-SCID and SCID-hu systems played an important role in HIV pathogenesis studies. However, despite many notable successes, several limitations still exist. They lack multi-lineage human hematopoiesis and a functional human immune system. These models primarily reflect an acute HIV infection with rapid CD4 T cell loss thus limiting pathogenesis studies to a short-term period. The new humanized Rag2-/-gamma c-/- mouse model (RAG-hu) created by intrahepatic injection of CD34 hematopoietic stem cells sustains long-term multi-lineage human hematopoiesis and is capable of mounting immune responses. Thus, this model shows considerable promise to study long-term in vivo HIV infection and pathogenesis. RESULTS: Here we demonstrate that RAG-hu mice produce human cell types permissive to HIV-1 infection and that they can be productively infected by HIV-1 ex vivo. To assess the capacity of these mice to sustain long-term infection in vivo, they were infected by either X4-tropic or R5-tropic HIV-1. Viral infection was assessed by PCR, co-culture, and in situ hybridization. Our results show that both X4 and R5 viruses are capable of infecting RAG-hu mice and that viremia lasts for at least 30 weeks. Moreover, HIV-1 infection leads to CD4 T cell depletion in peripheral blood and thymus, thus mimicking key aspects of HIV-1 pathogenesis. Additionally, a chimeric HIV-1 NL4-3 virus expressing a GFP reporter, although capable of causing viremia, failed to show CD4 T cell depletion possibly due to attenuation. CONCLUSION: The humanized RAG-hu mouse model, characterized by its capacity for sustained multi-lineage human hematopoiesis and immune response, can support productive HIV-1 infection. Both T cell and macrophage tropic HIV-1 strains can cause persistent infection of RAG-hu mice resulting in CD4 T cell loss. Prolonged viremia in the context of CD4 T cell depletion seen in this model mirrors the main features of HIV infection in the human. Thus, the RAG-hu mouse model of HIV-1 infection shows great promise for future in vivo pathogenesis studies, evaluation of new drug treatments, vaccines and novel gene therapy strategies. PMID- 17078893 TI - Ethnic differences in the time trend of female breast cancer incidence: Singapore, 1968-2002. AB - BACKGROUND: From 1968 to 2002, Singapore experienced an almost three-fold increase in breast cancer incidence. This increase appeared to be different across the three main ethnic groups: Chinese, Malays and Indians. This paper used age-period-cohort (APC) modelling, to determine the effects of age at diagnosis, calendar period, and birth cohort on breast cancer incidence for each ethnic group. METHODS: This study included all breast cancer cases (n = 15,269) in the three ethnic groups, reported to the Singapore Cancer Registry from 1968 to 2002 between the ages 25 to 79. Age-specific fertility rates from the Department of Statistics were used to explore the role of fertility. RESULTS: In the 1970s, Indian women had the highest age-standardized breast cancer but by the mid-1980s the highest rates were seen among the Chinese. Remarkable differences were seen in the age-specific incidence rates by ethnic groups. After age 49, the incidence rates for the Chinese and Malays leveled off whereas it continued to rise in the Indians. While our analyses provided some evidence that an age-drift model described the trend seen in the Indians, age-period-cohort model and age-cohort model had the best fit for the Chinese and Malays aged 25 to 79 respectively. Overall, Chinese and Malay women born in later cohorts were at increased risk of developing breast cancer relative to their counterparts in the earlier cohorts. The three ethnic groups experienced similar changes in their fertility in the 1970s, which likely explained much of the increase in their breast cancer incidence but not the ethnic differences. There was a stronger inverse association between total fertility rate and pre-menopausal breast cancer incidence in the Chinese and Malays than the Indians. CONCLUSION: The observed dissimilarity among ethnic groups suggests ethnic differences in exposure or response to certain risk factors. It is likely that longer and subtler differences in childbearing trends and other risk factors may further explain these ethnic differences. PMID- 17078892 TI - Interactions between IL-32 and tumor necrosis factor alpha contribute to the exacerbation of immune-inflammatory diseases. AB - IL-32 is a newly described cytokine in the human found to be an in vitro inducer of tumor necrosis factor alpha (TNFalpha). We examined the in vivo relationship between IL-32 and TNFalpha, and the pathologic role of IL-32 in the TNFalpha related diseases - arthritis and colitis. We demonstrated by quantitative PCR assay that IL-32 mRNA was expressed in the lymphoid tissues, and in stimulated peripheral T cells, monocytes, and B cells. Activated T cells were important for IL-32 mRNA expression in monocytes and B cells. Interestingly, TNFalpha reciprocally induced IL-32 mRNA expression in T cells, monocyte-derived dendritic cells, and synovial fibroblasts. Moreover, IL-32 mRNA expression was prominent in the synovial tissues of rheumatoid arthritis patients, especially in synovial infiltrated lymphocytes by in situ hybridization. To examine the in vivo relationship of IL-32 and TNFalpha, we prepared an overexpression model mouse of human IL-32beta (BM-hIL-32) by bone marrow transplantation. Splenocytes of BM-hIL 32 mice showed increased expression and secretion of TNFalpha, IL-1beta, and IL-6 especially in response to lipopolysaccharide stimulation. Moreover, serum TNFalpha concentration showed a clear increase in BM-hIL-32 mice. Cell-sorting analysis of splenocytes showed that the expression of TNFalpha was increased in resting F4/80+ macrophages, and the expression of TNFalpha, IL-1beta and IL-6 was increased in lipopolysaccharide-stimulated F4/80+ macrophages and CD11c+ dendritic cells. In fact, BM-hIL-32 mice showed exacerbation of collagen-antibody induced arthritis and trinitrobenzen sulfonic acid-induced colitis. In addition, the transfer of hIL-32beta-producing CD4+ T cells significantly exacerbated collagen-induced arthritis, and a TNFalpha blockade cancelled the exacerbating effects of hIL-32beta. We therefore conclude that IL-32 is closely associated with TNFalpha, and contributes to the exacerbation of TNFalpha-related inflammatory arthritis and colitis. PMID- 17078894 TI - Implementing referral guidelines: lessons from a negative outcome cluster randomised factorial trial in general practice. AB - BACKGROUND: Few patients with lower bowel symptoms who consult their general practitioner need a specialist opinion. However data from referred patients suggest that those who are referred would benefit from detailed assessment before referral. METHODS: A cluster randomised factorial trial. 44 general practices in North Trent, UK. Practices were offered either an electronic interactive referral pro forma, an educational outreach visit by a local colorectal surgeon, both or neither. The main outcome measure was the proportion of cases with severe diverticular disease, cancer or precancerous lesions and inflammatory bowel disease in those referred by each group. A secondary outcome was a referral letter quality score. Semi-structured interviews were conducted to identify key themes relating to the use of the software RESULTS: From 150 invitations, 44 practices were recruited with a total list size of 265,707. There were 716 consecutive referrals recorded over a six-month period, for which a diagnosis was available for 514. In the combined software arms 14% (37/261) had significant pathology, compared with 19% (49/253) in the non-software arms, relative risk 0.73 (95% CI: 0.46 to 1.15). In the combined educational outreach arms 15% (38/258) had significant pathology compared with 19% (48/256) in the non educational arms, relative risk 0.79 (95% CI: 0.50 to 1.24). Pro forma practices documented better assessment of patients at referral. CONCLUSION: There was a lack of evidence that either intervention increased the proportion of patients with organic pathology among those referred. The interactive software did improve the amount of information relayed in referral letters although we were unable to confirm if this made a significant difference to patients or their health care providers. The potential value of either intervention may have been diminished by their limited uptake within the context of a cluster randomised clinical trial. A number of lessons were learned in this trial of novel innovations. PMID- 17078895 TI - Facial palsy secondary to cholesteatoma: analysis of outcome following surgery. AB - Facial palsy is a rare presenting feature of cholesteatoma. Prompt treatment usually results in a good outcome, but if treatment is delayed the prognosis can be difficult to predict. We retrospectively reviewed 326 consecutive patients who had undergone temporal bone surgery for cholesteatoma. Eleven patients had presented with facial palsy, of whom eight had petrous apex involvement and three had disease confined to the middle-ear cleft. All patients with middle-ear disease were operated upon within two months of presentation, and all showed some recovery in facial function. The diagnosis had been delayed in all eight cases of apical disease, with four cases having had a long-standing total weakness. One case treated after seven months' partial weakness achieved a full recovery. In three cases of long-standing partial weakness, pre-operative facial function was preserved by maintaining the facial nerve in its normal anatomical location. Facial palsy associated with cholesteatoma should be treated as early as possible. Recovery can occur even if treatment is delayed for up to seven months. After this time, recovery is increasingly unlikely, but any residual function can be maintained and further deterioration prevented. PMID- 17078896 TI - Isolated inflammatory sphenoiditis with multiple unilateral cranial nerve palsies. AB - Isolated sphenoidits is a rare entity that often presents with vague, non specific symptoms. We present the case of a 36-year-old Middle Eastern man, who developed headache and a painful right eye. A diagnosis of acute sphenoiditis was made. Shortly afterwards, he developed diplopia due to isolated abducent nerve involvement. Within two months, the extent of cranial nerve involvement had increased to include cranial nerves II, III, and V. Subsequently, this was treated by functional endoscopic sinus surgical drainage and biopsy. Histology revealed inflammatory changes. The patient made a dramatic recovery post operatively, with resolution in all symptoms. PMID- 17078897 TI - Quality of life outcome analysis in patients undergoing submandibular duct repositioning surgery for sialorrhoea. AB - Bilateral submandibular duct transposition is one of several surgical methods described to control sialorrhoea in the paediatric patient. The purpose of this study is to assess the effect of submandibular duct repositioning surgery on the quality of life of children using the Glasgow Benefit Inventory outcome measure. Consecutive children who underwent submandibular duct repositioning surgery were invited to participate in the study. The parents or guardians of children who gave consent were interviewed. The carer's opinion of the change in salivation, responses to the items in the Glasgow Benefit Inventory score and report of the complications were recorded. Nine children aged two and a half to 16 years were studied. The mean follow-up time was four years. The mean Glasgow Benefit Inventory score of +33 related to the procedure demonstrates that submandibular duct repositioning leads to a substantial measurable improvement in the quality of life (QoL) of the drooling child. PMID- 17078898 TI - Early results of titanium ossiculoplasty using the Kurz titanium prosthesis--a UK perspective. AB - Titanium has been shown to be a highly biocompatible material and recently interest has been raised by its use as a prosthetic material in the ear. Our objective was to prospectively assess the early results of titanium partial and total ossicular replacement prostheses in chronic ear disease. Thirty-three consecutive patients (20 titanium partials and 13 total ossicular replacement prostheses) were analysed and data from pre and post-operative pure tone audiograms were collected and compared at six months over a four frequency range (0.5-4 kHz) as well as using the American Academy of Otolaryngology-Head and Neck Surgery criteria (0.5-3 kHz). Closure of the air-bone gap and improvement in air conduction at six months was also calculated. 69% of patients obtained an air bone gap of < or = 20 dB, with titanium partial ossicular replacement prostheses (85 per cent) doing better than total ossicular replacement prostheses (46 per cent). There have been no cases of sensorineural hearing loss or extrusion. In conclusion, titanium prostheses are easy to use, allow good visibility and have shown good short-term results in our hands. PMID- 17078899 TI - A historical review of head and neck cancer in celebrities. AB - INTRODUCTION: The illnesses of celebrity patients always receive more attention from the general public than those of ordinary patients. With regard to cancer, this fact has helped to spread information about the four major malignancies: breast cancer, prostatic cancer, lung cancer and colorectal cancer. Head and neck cancer, on the other hand, is still not well recognised by the lay public, although the risk factors are similar to those of lung cancer. It was the objective of this analysis to identify cases of celebrity patients, the description of which could help to increase awareness of head and neck cancer, its symptoms and risk factors. METHODS: The Internet and medical literature databases were searched for celebrity patients who had suffered from head and neck cancer. RESULTS: The search revealed numerous famous head and neck cancer patients. However, only seven cases were documented well in the medical literature. Among the identified persons were one emperor, two United States presidents, a legendary composer, a world-renowned medical doctor, an outstanding athlete and an extraordinary entertainer. In spite of their exclusive position in society, these patients did not have a better prognosis compared with ordinary patients of their time. Only two of the group experienced long term survival and only one was cured. None of these influential figures used their influence to fund research or to promote knowledge about their respective diseases. CONCLUSION: The identified cases could help increase public awareness of head and neck cancer. Similar to activities in other oncologic fields, current celebrity head and neck cancer patients should be encouraged to discuss their diseases openly, which could have a positive effect on public health. PMID- 17078900 TI - A rare case of nasal cysticercosis mimicking a nasal dermoid. AB - Cysticercosis, caused by infestation with the pork tapeworm Taenia solium, is known to commonly involve the brain, muscles and subcutaneous tissue. Diagnosing cysticercosis can be difficult as its clinical presentation is usually non specific. When involving subcutaneous tissue, it can present as firm, non-tender, solitary or multiple nodules. We report a case of subcutaneous cysticercosis at the nasion mimicking a nasal dermoid. PMID- 17078901 TI - Student-selected components: bringing more ENT into the undergraduate curriculum. AB - Exposure to otolaryngology is currently minimal in the UK undergraduate medical curriculum. This may lead to difficulties in attracting graduates into higher ENT surgical training and in ensuring a reasonable standard of ENT knowledge amongst primary care practitioners. A recent innovation, of which many ENT units may be unaware, is the introduction to the undergraduate curriculum of 'student-selected components'. Like the traditional elective, this allows students to undertake an attachment to a speciality and department of their choice. Units which do not regularly teach medical students but which have a welcoming and enthusiastic approach to undergraduate training may well be ideal hosts. This paper introduces the concepts underlying student-selected components, outlines the preparation required and offers a template for such an attachment, for which ENT is ideally suited. PMID- 17078902 TI - A rare anatomical variation of the spinal accessory nerve. AB - Considerable inter- and intra-individual anatomical differences can be observed during neck dissections. In this case report, we describe an unusual relationship of the accessory nerve to the internal jugular vein. PMID- 17078903 TI - Effects of parasitization by Varroa destructor on survivorship and physiological traits of Apis mellifera in correlation with viral incidence and microbial challenge. AB - Varroa mites (Varroa destructor) are serious ectoparasites of honey bees (Apis mellifera). This research addresses the impact of varroa mites on survivorship, viral incidence, and physiological traits of newly-emerged worker bees. RT-PCR confirmed our previous finding that varroa parasitization was linked to high levels of deformed wing virus (DWV). In non-treatment bees, varroa parasitization combined with increased viral levels altered survivorship curves from long survival to shorter-survival types. After challenge with live Escherichia coli, the survivorship of mite-parasitized bees was significantly lower than mite-free bees. Deformed-wing, mite-parasitized bees died on average within 1 day, even without E. coli challenge. This was correlated with the absence of an important enzyme activity in insect immunity, phenol oxidase, lacking even in those bees challenged with immuno-elicitors. The lack of inducible phenol oxidase activity indicated that the bee immune system is not fully competent upon adult emergence. Varroa parasitism also significantly reduced body weight of the parasitized bees, but body weight was not significantly correlated with the survivorship of mite parasitized bees. Our research indicates that the combination of mite parasitization, the interaction of DWV and microbes, and a developmental immune incompetency attribute to decreased worker survivorship and have a negative impact on colony fitness. PMID- 17078904 TI - Anti-filarial activity of novel formulations of albendazole against experimental brugian filariasis. AB - The study was aimed at developing better orally active albendazole (ALB) formulations. Six formulations (ALB-1 to ALB-6) were prepared and tested against Brugia malayi in Mastomys coucha and jird (Meriones unguiculatus) at 200 mg/kg, orally, for 5 consecutive days. The anti-filarial efficacy was assessed against microfilariae (mf), adult worms and female reproductive potential. Three of the 6 ALB formulations showed greatly improved female worm sterilizing potential (ALB 1: 90%; ALB-3: 63%; ALB-4: 77% of untreated control) in B. malayi - M. coucha model. Sterilization efficacy of ALB-1 was also better than that shown by pure ALB (P<0.001) or its marketed tablet formulation, Zentel (P<0.01), while that of ALB-4 was better than pure-ALB (P<0.05). The activity of ALB-3, pure-ALB and Zentel was, however, comparable. ALB-1 also showed late microfilaricidal activity with a maximum of 78% fall in microfilarial count. In contrast, neither the pure ALB nor Zentel showed any microfilaricidal activity. In the jird - B. malayi model, ALB-1 and ALB-4 showed marginal sterilizing efficacy whereas pure ALB or Zentel were ineffective. In conclusion the anti-filarial efficacy of ALB-1 was found to be superior to pure-ALB or Zentel. PMID- 17078905 TI - Determinants of population biology of the chewing louse Brueelia apiastri (Mallophaga, Philopteridae) on the European bee-eater (Merops apiaster). AB - In this study we examine the population biology of Brueelia apiastri, a chewing louse living on the European bee-eater (Merops apiaster). We investigate the relationships between parasite intensity of infestation, sex ratio, reproductive output, parasite size and their environment i.e. the morphology, condition, age and sex of the host. Chewing lice were collected, their sex and age (developmental stage) identified and parasite body size determined as a measure of parasite condition (larger individuals consume larger meals and larger females may produce larger clutches). The data show that there is variation in intensity as well as body size of B. apiastri between individual bee-eaters and this variation is independent of the sex of the birds. However, size, condition and age of the birds seem to influence the infestation rates with B. apiastri. The study suggested size-dependent depredation, since more, smaller chewing lice (usually nymphs) living on birds in better condition and birds having longer bills. Furthermore, more male chewing lice (males are smaller than females) live on older birds. Intraspecific competition between parasites seems to have a negative effect on female but not male body size but this result could be also explained by size-dependent depredation. PMID- 17078906 TI - Timing of surgery in aortic stenosis. AB - Aortic valve stenosis (AVS) usually results from three distinct processes (degenerative-calcific, rheumatic, and congenital), with a final common pathway of significant aortic outflow tract obstruction. The stenotic lesion tends to progress slowly, but once symptoms develop clinical deterioration can ensue rapidly. Chest pain, dyspnea, and syncope are the most common symptoms of significant AVS. Detection of symptoms, subtle or obvious, is critical to the management of AVS because their presence portends a worse overall prognosis and is an indication for intervention. There are several special clinical scenarios that require added consideration, including individuals with concomitant coronary artery disease, the presence of a relatively small transvalvular pressure gradient in the setting of low cardiac output (so-called low-gradient AVS), and elderly with severe AVS. Surgical aortic valve replacement (AVR) is the mainstay treatment for relief of obstruction in patients with symptomatic AVS. Percutaneous balloon valvuloplasty is reserved for the small minority of patients who are not surgical candidates and is associated with a high restenosis rate. Percutaneous AVR is a new technology that is being tested in a few select centers on patients who are not operative candidates. PMID- 17078907 TI - Role of vasodilators in regurgitant valve disease. AB - Vasodilator therapy is designed to reduce regurgitant volume and improve left ventricular function. Acute administration reduces vascular resistance and decreases regurgitant volume and left ventricular filling pressure. These effects may be clinically useful in acute regurgitations, but less consistent results have been reported in long-term therapy. In chronic mitral functional regurgitation, vasodilator therapy has proved to have clinical or prognostic benefit only when heart failure or poor ventricular function is present. The indication of vasodilator treatment in aortic regurgitation has raised significant controversy. Several studies with small series have shown beneficial effects on regurgitant volume, ejection fraction, and mass of the left ventricle. Nevertheless, in the only two randomized long-term follow-up studies, results differed completely. In our experience, both nifedipine and enalapril failed to reduce the need for valvular surgery or show benefits in echocardiographic parameters. Vasodilator therapy would be indicated only in patients with severe aortic regurgitation and systemic hypertension, or when surgery is contraindicated. PMID- 17078908 TI - Selection of prosthetic heart valves. AB - Cardiovascular surgery for the repair or replacement of diseased heart valves has continually improved since its introduction in the early 1960s. Despite advances in prosthetic heart valve design, to date there is no valve that is comparable to the native human valve with respect to durability, risk of thrombosis, and overall hemodynamic function. Although bioprosthetic devices are similar to the native valve with respect to thrombogenicity, durability is a significant concern, particularly in younger patients. Approximately 45% of implanted bioprosthetic valves fail at 10 years. In contrast, mechanical prostheses have a significantly lower incidence of structural failure, with an implantation life of greater than 20 years, and are thus more often used for patients under the age of 65. Unfortunately, significant hemodynamic and thrombotic issues have yet to be resolved with the latest generation of mechanical valves. Thus, careful analysis of patient factors and valve-related complications must be considered when treatment of heart valve disease is offered to the patient. The purpose of this review is to discuss the current recommendations for surgical intervention for heart valve disease. PMID- 17078909 TI - Imaging and quantifying valvular heart disease using magnetic resonance techniques. AB - Echocardiography remains the cornerstone of noninvasive valvular heart disease evaluation. There are instances where MRI can be of use. Aside from the obvious advantage where limited acoustic windows are present, cardiac magnetic resonance (CMR) allows for imaging in any desired plane, and advantage can be taken of the ability to align with any regurgitant or stenotic flow jet. The high spatial resolution and contrast allow for accurate detail of valvular anatomy, but it must be remembered that the images represent a composite of eight to 12 heart cycles. For visualizing multiple valvular abnormalities simultaneously, cardiac MRI has a distinct advantage. Finally, a CMR valvular examination can be combined with accurate assessments of left and right ventricular function, myocardial stress perfusion imaging, and detailed viability determinations in a single examination. This provides a comprehensive presurgical evaluation of cardiac physiology. PMID- 17078910 TI - Approach to the patient with bicuspid aortic valve and ascending aorta aneurysm. AB - Bicuspid aortic valve (BAV) disease is a common congenital heart valve abnormality accounting for a large number of valve replacements in the United States. Although still incompletely understood, the natural history of BAV disease is severe aortic stenosis and associated ascending aortic dilatation. In addition to the increased risk of endocarditis, aortic dissection and severe aortic valve dysfunction are responsible for most fatal complications. Thus, early and precise recognition of this condition is mandatory. The new American College of Cardiology/American Heart Association recommendations highlight the role of MRI and CT as complimentary tools to echocardiography for the diagnosis and surveillance of the morphology of the aortic valve and ascending aorta. Moreover, better understanding of the cellular mechanisms, including inflammation, bone formation, atherosclerotic-like processes, and aortic wall abnormalities, as well as the heritability and genetic predisposition for the disease, will define the potential for targeted medical therapies in the future. Currently, the treatment of this condition is primarily surgical. Although combined valve and ascending aorta replacement has been the most common surgical approach in the past, the increased cumulative risk of thrombotic and embolic events among these young patients has led to more conservative approaches. Several valve-sparing approaches with comparable mid-term results compared with the classic procedures have recently been reported. However, longer follow-up studies will be helpful to better define the advantages of these new surgical options. After a quick overview of the natural history of the BAV, this article provides an updated approximation of the current knowledge of the pathophysiology as well as the recommendations for the management and treatment of this disease. PMID- 17078911 TI - Current treatment in cardiac amyloidosis. AB - Involvement of the heart is a common finding in amyloidosis. The heart is usually infiltrated by amyloid fibrils in primary amyloidosis and age-related forms of amyloidosis, less commonly in transthyretin familial amyloidosis, and rarely in secondary amyloidosis. The most common clinical presentation is restrictive cardiomyopathy with right-sided heart failure. The second most frequent presentation is congestive heart failure due to systolic dysfunction, followed by arrhythmias and orthostatic hypotension. The diagnosis of amyloidosis requires tissue sample confirmation; at present, Congo red staining in polarized light is the diagnostic method of choice. The characterization of protein fibril type by immunohistochemistry or biochemistry is essential for patient prognosis and treatment. The therapeutic approach consists of specific treatment of amyloidosis and supportive treatment for cardiac-related symptoms. The treatment depends on the type of amyloidosis and the stage of disease. The mainstay of supportive treatment of cardiac failure is diuretic therapy. Primary amyloidosis treatment protocol includes melphalan and prednisone chemotherapy. Heart transplantation is only a palliative treatment. Stem cell transplantation is an emerging treatment alternative. Combination therapy of melphalan and stem cell transplantation has been shown to be a promising treatment strategy. Secondary amyloidosis requires aggressive treatment of the associated inflammatory and neoplastic process. Age related (senile) amyloidosis benefits from supportive cardiac treatment when applicable. Transthyretin amyloidosis, the most common cardiac hereditary amyloidosis, is treated by liver or combined liver-heart transplantation. New therapies based on chemical and immunologic reaction with amyloid or its precursor are under intensive development. PMID- 17078912 TI - Management after childhood repair of tetralogy of fallot. AB - The four characteristic findings in tetralogy of Fallot (TOF) include the following: 1) a malaligned ventricular septal defect; 2) right ventricular outflow and/or pulmonary valve/artery stenosis or atresia; 3) a dextraposed overriding aorta; and 4) right ventricular hypertrophy. This article focuses solely on the outcomes and treatment of surgically corrected patients born with TOF. In the modern era, early surgical repair consisting of ventricular septal defect closure and alleviation of right ventricular outflow obstruction have gained favor over early palliation with an aortopulmonary shunt followed by intracardiac repair. Surgical outcomes are excellent and dramatically improve prognosis. However, these patients are not "cured" and are at significant risk of developing subsequent electrical and hemodynamic problems. An operated patient with TOF should be evaluated at regular intervals by a cardiologist trained in congenital heart disease; any symptoms suggestive of hemodynamic or electrical compromise should spur further investigation. Advances in imaging, medical therapy, electrophysiology, device/resynchronization therapy, and percutaneous intervention provide the clinician with a number of therapeutic options. Surgical techniques have changed significantly since the early intracardiac repairs of the 1960s and 1970s. The deleterious hemodynamic and electrical effects of pulmonary regurgitation and ventriculotomy scars have spurred efforts to ensure pulmonary valvular competence and minimize the extent of ventricular incisions. Since Blalock and Taussig's first palliative shunt in 1945, the survival and quality of life for patients with TOF has improved dramatically; this is one of the great accomplishments for cardiovascular medicine in the 20th century. The 21st century promises further success with the advent of a myriad of technologic advancements. PMID- 17078913 TI - Cell-based therapies after myocardial injury. AB - Recent translational research into the emerging field of cardiac cell therapy has paved the way for novel clinical treatment strategies. However, neither the ideal source and type of cell nor the critical quantity and mode of application have yet been defined. In patients who have undergone acute myocardial infarction, several cell-based approaches are currently being evaluated, such as intracoronary delivery of autologous mononuclear bone marrow cells or enriched hematopoietic progenitor cell products; systemic cytokine stimulation with release of bone marrow progenitor cells into the systemic circulation; and both intravenous and intracoronary delivery of allogenic marrow stroma cell-derived cells. There are potentially encouraging data for each of these strategies, based to date on small cohorts with conflicting or equivocal recovery of function. Taken together, it is too early to consider cell therapy for heart disease to be effective. Future setbacks are likely, but both clinicians and basic scientists will eventually introduce more potent cell-based strategies into the clinical arena. PMID- 17078914 TI - Bicycle helmets and brain injury. AB - This paper replies to criticism by Cummings et al. [Cummings, P., Rivara, F.P., Thompson, D.C., Thompson, R.S., 2006. Accid. Anal. Prev. 38, 636-643] of an article [Curnow, W.J., 2005. The Cochrane Collaboration and bicycle helmets. Accid. Anal. Prev. 37, 569-573] disputing a conclusion of a Cochrane Collaboration review, namely, that it establishes scientific evidence that all types of standard bicycle helmet protect against injury to the brain. In response to the conclusion of Cummings that the review's case-control studies provide such evidence, I explain that their design is inadequate to do this. PMID- 17078915 TI - Infants' imitation of goal-directed actions: the role of movements and action effects. AB - This paper reviews studies on infants' imitation of goal-directed actions in the first two years of life. Special emphasis is given to the role of the two observable components of an action, that is, the movement and the action effects, on infants' replication of target actions. The reviewed studies provide evidence that infants benefit most from a full demonstration of both movements and effects. If movements are demonstrated in isolation, infants may encode this information, but they preferentially reproduce actions that lead to salient effects. If action effects are presented in isolation, infants younger than 19 months usually fail to emulate the unseen movements that would be necessary to produce these effects. Infants' ability to predict action effects or to infer unseen movements from incomplete demonstrations improves substantially at the end of the second year of life. It is concluded that the capability to learn relations between movements and action effects by observation, and the knowledge about movement-effect relations acquired so far, may be important factors underlying the developmental changes in infants' imitation of goal-directed actions. PMID- 17078916 TI - Comparison of the in vitro drug sensitivity of Trypanosoma brucei gambiense strains from West and Central Africa isolated in the periods 1960-1995 and 1999 2004. AB - The situation of human African trypanosomiasis remains serious with one of the main threats being the increasing number of relapses or treatment failures after melarsoprol treatment. In order to investigate and to compare drug sensitivities of trypanosomes isolated at different time periods and in different locations, two sets of Trypanosoma brucei gambiense strains were used. One set was isolated in the time period 1960-1981 and the other one in 1995-2004 from different locations of West and Central Africa. These isolates were not selected based on the treatment outcome but on availability. The drug sensitivity profile for all available drugs in use and the diamidine compound DB75 was established. IC(50) values were not significantly different between the "old" and "new" stocks. No indications for emerging drug resistance to any drug could be observed. The results indicate a relative stability of in vitro sensitivity of T. b. gambiense to trypanocidal drugs in space (West and Central Africa) and time (1960-2004). PMID- 17078917 TI - Distribution of 28.5 kDa antigen in the tegument of adult Fasciola gigantica. AB - Monoclonal antibody (MoAb) specific to 28.5 kDa tegumental antigen (TA) was used to localize this antigen in various tissues of adult Fasciola gigantica by means of indirect immunofluorescence, immunoperoxidase and immunogold techniques. The indirect immunofluorescence and immunoperoxidase detections revealed that this antigen was concentrated in the tegument particularly in its outer rim, tegumental cells and their processes, epithelial linings of the oral sucker and the proximal part of digestive tract. It was also detected at a moderate concentration in spermatogenic cells in the testes, cells of Mehlis' gland, oocytes within the ovary, and ovum within the egg of adult parasites. At TEM level, the immunogold detection showed deposit of gold particles specifically in G(2) tegumental granules and on the surface membrane. Thus, this antigen is expressed in the tegument and associated structures of adult parasites, and it could be a major component of the G(2) granules which are shown to fuse with the surface membrane and contribute material to replace the casted-off membrane. This process is a part of membrane turnover that prevents the parasite from being attacked by the host immune effector cells. PMID- 17078918 TI - Acanthamoeba isolates belonging to T1, T2, T3, T4 and T7 genotypes from environmental freshwater samples in the Nile Delta region, Egypt. AB - The free-living amoebae of the genus Acanthamoeba include non-pathogenic and pathogenic species and has been recently classified into 15 different genotypes, T1-T15. In this study, a survey was conducted in order to determine the presence and pathogenic potential of free-living amoebae of Acanthamoeba genus in freshwater sources associated with human activities in the Nile Delta region, Egypt. Identification of Acanthamoeba was based on the morphology of cyst and trophozoite forms and PCR amplification with a genus specific primer pair. The pathogenic potential of Acanthamoeba isolates was characterized using temperature and osmotolerance assays and PCR reactions with two primer pairs specific to Acanthamoeba pathogenesis. Isolates genotypes were also determined after ribosomal DNA sequencing. These data revealed that isolates belong to T1, T2, T3, T4 and T7 genotypes. As expected, T4 isolates exhibited the most pathogenic traits and were osmotolerant, temperature tolerant and expressed extracellular serine proteases. This is the first report presenting environmental distribution of Acanthamoeba genotypes in Egypt. PMID- 17078919 TI - Integration of an on-line protein digestion microreactor to a nanoelectrospray emitter for peptide mapping. AB - A method for integrating nanoelectrospray mass spectrometry with a microreactor for on-line digestion and fast peptide mass mapping from dilute protein samples is presented. Fused silica capillaries (i.d. 50 microm, o.d. 360 microm) are employed as the digestion microreactor and the nanoelectrospray emitter by immobilizing trypsin onto the surface of the inner wall of the fused silica capillary tubing. The procedure is demonstrated using solutions of 1pmol/mul angiotensin II, cytochrome c, hemoglobin, and beta-casein. Because the inner walls of the capillaries are modified by covalent chemical bonds, the adsorption of peptides and proteins to the inner walls of the capillaries is suppressed. This procedure was performed with solutions as dilute as 1fmol/mul (1nM) cytochrome c. This method shows generation of tryptic peptides with sequence coverage up to 90% within minutes; trypsin autolysis products are not detected. In addition, the immobilized enzyme can be cleaned easily, enabling the microreactor to be reused for nanoelectrospray. PMID- 17078920 TI - Evaluation of orientation and environmental factors on the blast hazards to bomb suit wearers. AB - This paper discusses the use of 3-D computational fluid dynamics (CFD) software for simulation of explosive detonations against the wearer of a typical bomb suit. The focus is to demonstrate the utility of the model to assess the differential overpressure loads on the various bomb suit components protecting critical body parts. Since overpressures can vary significantly depending on the position and orientation of the wearer, simulations are performed for a range of orientations including kneeling and standing at common standoff distances. Overpressure loads on the head, neck, and torso regions are predicted for each orientation, capturing ground and other surface reflections that can enhance effects of the blast. This is not normally observed during tests of individual bomb suit components. Direct correlations between the suit orientation and environmental factors to load enhancements are documented, and an effort to address probable injury is made. PMID- 17078921 TI - Heme crevice disorder after sixth ligand displacement in the cytochrome c-551 family. AB - 1H NMR and visible absorption spectroscopy were used to monitor sixth ligand methionine displacement reactions in four members of the ferricytochrome c-551 family from Pseudomonas aeruginosa, Pseudomonas stutzeri, Pseudomonas stutzeri substrain ZoBell, and Nitrosomonas europae. Potassium cyanide displaces the methionine ligand with very modest changes in the visible spectra, but profound changes in the NMR spectra. The initial product formed kinetically, designated complex I, changes with time and/or heating to a more thermodynamically favored product termed complex II. Spectra indicate that both I and II are actually a family of closely related conformational isomers. Low temperature NMR spectra of complex II indicate that some of the isomers are in chemical exchange on the NMR time scale. High pH also displaces the methionine ligand in a manner similar to the well-known alkaline transition of mitochondrial cytochrome c. However, the reaction occurs at higher pH values and over a narrower pH range for the c-551 family, and the transition pH range is different for the different proteins studied. The final alkaline forms also show peak widths and a number of peaks indicative of multiple conformational isomers. PMID- 17078922 TI - A conserved molecular action of native and recombinant Epap-1 in inhibition of HIV-1 gp120 mediated viral entry. AB - The early expression of Epap-1 (early pregnancy associated protein), a 90 kDa anti-HIV-1 active glycoprotein, in the first trimester placental tissue suggests that it is one of the innate immune factors/proteins protecting the fetus from HIV infections. In the present investigation, we have cloned and expressed Epap-1 in bacterial and baculovirus expression systems. The recombinant Epap-1 as well as native Epap-1 shows a conserved molecular mode of action. These proteins exhibit significant antiviral activity and inhibit the cell fusion reaction between gp120 expressing HeLa (HL2/3) cells and T cell line (SupT1). Further, the rhodamine labeled Epap-1 specifically bound to gp120 expressed on the surface of HL2/3 cells during fusion reaction thereby inhibiting viral entry. Analysis of the interacting gp120 epitopes revealed that Epap-1 binds specifically to epitopes of gp120, recognizing constant-5 (C5) region and the variable-3 (V3) epitope of gp120 expressed on HL2/3 cells; It exhibits specific interaction with C5 region of cell-free virus in four HIV-1 isolates suggesting that the molecular interaction of Epap-1 is specific and is highly conserved in binding to gp120 leading to inhibition of viral entry. Epap-1 can thus be a very efficient natural protection mechanism against cell-free and cell-associated viral infections during early pregnancy. PMID- 17078923 TI - Redox reactions of dehydroepiandrosterone and its metabolites in differentiating 3T3-L1 adipocytes: A liquid chromatographic-mass spectrometric study. AB - Dehydroepiandrosterone is known to depress fatty acid formation in differentiating 3T3-L1 adipocytes. The metabolism of dehydroepiandrosterone and four of its natural metabolites in differentiating adipocytes was studied by liquid chromatography-mass spectrometry. Adipocytes rapidly converted dehydroepiandrosterone to androst-5-ene-3beta,17beta-diol. 7alpha-Hydroxy-DHEA was interconverted with 7-oxo-DHEA and 7beta-hydroxy-DHEA and the corresponding 17beta reduced products. Dehydroepiandrosterone and its derivatives were detected only in the culture medium suggesting that dehydroepiandrosterone is metabolized via enzymes located in close proximity to, or that are integral parts of the cell membrane. Alternatively, there may be efficient mechanisms at play for extrusion of the steroids to the aqueous media rather than being retained in the lipid-rich cell. An interesting aspect of the study was finding androstenediol as the major metabolite of dehydroepiandrosterone. Androst-5-ene-3beta,17beta-diol has been implicated in prostate cancer. The contribution of adipose cells to the circulating supply of androst-5-ene-3beta,17beta-diol may therefore be considered in managing prostate cancer. PMID- 17078924 TI - A unique insertion/duplication in the VDR gene that truncates the VDR causing hereditary 1,25-dihydroxyvitamin D-resistant rickets without alopecia. AB - Hereditary vitamin D resistant rickets (HVDRR) is caused by mutations in the vitamin D receptor (VDR). Here we describe a patient with HVDRR who also exhibited some hypotrichosis of the scalp but otherwise had normal hair and skin. A 102 bp insertion/duplication was found in the VDR gene that introduced a premature stop (Y401X). The patient's fibroblasts expressed the truncated VDR, but were resistant to 1,25(OH)2D3. The truncated VDR weakly bound [3H] 1,25(OH)2D3 but was able to heterodimerize with RXR, bind to DNA and interact with the corepressor hairless (HR). However, the truncated VDR failed to bind coactivators and was transactivation defective. Since the patient did not have alopecia or papular lesions of the skin generally found in patients with premature stop mutations this suggests that this distally truncated VDR can still regulate the hair cycle and epidermal differentiation possibly through its interactions with RXR and HR to suppress gene transactivation. PMID- 17078925 TI - Lysophospholipid receptors: signalling, pharmacology and regulation by lysophospholipid metabolism. AB - The lysophospholipids, sphingosine-1-phosphate (S1P), lysophosphatidic acid (LPA), sphingosylphosphorylcholine (SPC) and lysophosphatidylcholine (LPC), activate diverse groups of G-protein-coupled receptors that are widely expressed and regulate decisive cellular functions. Receptors of the endothelial differentiation gene family are activated by S1P (S1P(1-5)) or LPA (LPA(1-3)); two more distantly related receptors are activated by LPA (LPA(4/5)); the GPR(3/6/12) receptors have a high constitutive activity but are further activated by S1P and/or SPC; and receptors of the OGR1 cluster (OGR1, GPR4, G2A, TDAG8) appear to be activated by SPC, LPC, psychosine and/or protons. G-protein-coupled lysophospholipid receptors regulate cellular Ca(2+) homoeostasis and the cytoskeleton, proliferation and survival, migration and adhesion. They have been implicated in development, regulation of the cardiovascular, immune and nervous systems, inflammation, arteriosclerosis and cancer. The availability of S1P and LPA at their G-protein-coupled receptors is regulated by enzymes that generate or metabolize these lysophospholipids, and localization plays an important role in this process. Besides FTY720, which is phosphorylated by sphingosine kinase-2 and then acts on four of the five S1P receptors of the endothelial differentiation gene family, other compounds have been identified that interact with more ore less selectivity with lysophospholipid receptors. PMID- 17078927 TI - The Children's Automatic Thoughts Scale in a clinical sample: psychometric properties and clinical utility. AB - The aim of this study was to examine the psychometric properties and clinical utility of the Children's Automatic Thoughts Scale (CATS; Schniering, C. A., & Rapee, R. M. (2002). Development and validation of a measure of children's automatic thoughts: The Children's Automatic Thoughts Scale. Behaviour Research and Therapy, 40, 1091-1109) in a large sample of anxious youth. The participants were 891 referred children and adolescents. Participants completed the CATS and a wide range of symptom measures, and were assessed via a structured diagnostic interview. Previous community-based psychometric properties were confirmed. The scale was highly sensitive to treatment change, and showed evidence of cognitive specificity with reductions in threat and failure beliefs, but not in hostility beliefs following treatment. The CATS demonstrated good convergent validity with related anxiety and depression scales, and moderate discriminant validity was found across anxious, anxious-depressed and anxious-oppositional groups. Implications for the assessment of child anxiety, and difficulties around children "faking good" on anxiety measures are discussed. PMID- 17078928 TI - A novel meta-cleavage product hydrolase from Flavobacterium sp. ATCC27551. AB - The organophosphate degrading (opd) gene cluster of plasmid pPDL2 of Flavobacterium sp. ATCC27551 contains a novel open-reading frame, orf243. This was predicted to encode an alpha/beta hydrolase distantly related to the meta fission product (MFP) hydrolases such as XylF, PhnD, and CumD. By homology modeling Orf243 has most of the structural features of MFP hydrolases including the characteristic active site catalytic triad. The purified protein (designated MfhA) is a homotetramer and shows similar affinity for 2-hydroxy-6-oxohepta-2,4 dienoate (HOHD), 2-hydroxymuconic semialdehyde (HMSA), and 2-hydroxy-5 methylmuconic semialdehyde (HMMSA), the meta-fission products of 3-methyl catechol, catechol, and 4-methyl catechol. The unique catalytic properties of MfhA and the presence near its structural gene of cis-elements required for transposition suggest that mfhA has evolved towards encoding a common hydrolase that can act on meta-fission products containing either aldehyde or ketone groups. PMID- 17078929 TI - Jak2 and Tyk2 are necessary for lineage-specific differentiation, but not for the maintenance of self-renewal of mouse embryonic stem cells. AB - As the LIF-induced Jak1/STAT3 pathway has been reported to play a crucial role in self-renewal of mESCs, we sought to determine if Jak2, which is also expressed in mESCs, might also be involved in the pathway. By employing an RNAi strategy, we established both Jak2 and Jak2/Tyk2 knockdown mESC clones. Both Jak2 and Jak2/Tyk2 knockdown clones maintained the undifferentiated state as wild-type controls, even in a very low concentration of LIF. However, we observed not only faster onset of differentiation but also differential expression of tissue specific lineage genes for ectodermal and mesodermal, but not endodermal origins from embryoid bodies generated from both types of knockdown clones compared to the wild-type. Furthermore, the reduced level of Jak2 caused differentiation of mESCs in the presence of LIF when the Wnt pathway was activated by LiCl treatment. Taken together, we demonstrated that Jak2 and Tyk2 are not involved in LIF-induced STAT3 pathway for self-renewal of mESCs, but play a role in early lineage decision of mESCs to various differentiated cell types. PMID- 17078930 TI - STAM-AMSH interaction facilitates the deubiquitination activity in the C-terminal AMSH. AB - Signal transducing adaptor molecule (STAM) complexed with hepatocyte growth factor regulated tyrosine kinase substrate (Hrs) works on sorting of cargo proteins in multivesicular body (MVB) pathway. Associated molecule with SH3 domain of STAM (AMSH), a zinc-containing ubiquitin isopeptidase, is thought to play a role in regulation of ubiquitin-mediated degradation by binding to STAM. We have found that AMSH requires the conformation of Px(V/I)(D/N)RxxKP sequence to bind SH3 domain of STAM with approximately 7 microM affinity, and that the isolated C-terminal domain of AMSH contains the isopeptidase activity. Deubiquitination by AMSH was assisted when ubiquitins were bound to STAM which can bind to AMSH simultaneously. With the specificity toward K63-linked ubiquitins, this facilitated ubiquitin processing activity of AMSH may imply a distinct regulatory mechanism for sorting and degradation through STAM binding. PMID- 17078926 TI - G protein-coupled receptors in major psychiatric disorders. AB - Although the molecular mechanisms underlying psychiatric illnesses such as depression, bipolar disorder and schizophrenia remain incompletely understood, there is increasing clinical, pharmacologic, and genetic evidence that G protein coupled receptors (GPCRs) play critical roles in these disorders and their treatments. This perspectives paper reviews and synthesizes the available data. Dysfunction of multiple neurotransmitter and neuropeptide GPCRs in frontal cortex and limbic-related regions, such as the hippocampus, hypothalamus and brainstem, likely underlies the complex clinical picture that includes cognitive, perceptual, affective and motoric symptoms. The future development of novel agents targeting GPCR signaling cascades remains an exciting prospect for patients refractory to existing therapeutics. PMID- 17078931 TI - Constitutive homo- and hetero-oligomerization of TbetaRII-B, an alternatively spliced variant of the mouse TGF-beta type II receptor. AB - Transforming growth factor (TGF)-beta ligands signal through transmembrane type I and type II serine/threonine kinase receptors, which form heteromeric signalling complexes upon ligand binding. Type II TGF-beta receptors (TbetaRII) are reported to exist as homodimers at the cell surface, but the oligomerization pattern and dynamics of TbetaRII splice variants in live cells has not been demonstrated thus far. Using co-immunoprecipitation and bioluminescence resonance energy transfer (BRET), we demonstrate that the mouse TbetaRII receptor splice variant TbetaRII-B is capable of forming ligand-independent homodimers and heterodimers with TbetaRII. The homomeric interaction of mouse (m)TbetaRII-B isoforms, however, is less robust than the heteromeric interactions of mTbetaRII-B with wild-type TbetaRII, which indicates that these receptors may be more likely to heterodimerize when both receptors are expressed. Moreover, we demonstrate that mTbetaRII-B is a signalling receptor with ubiquitous tissue expression. Our study thus demonstrates previously unappreciated complex formation of TGF-beta type II receptors, and suggests that mTbetaRII-B can direct TGF-beta-induced signalling in vitro and in vivo. PMID- 17078932 TI - Oral administration of phenolic antidiarrheic ingredients prevents ovariectomy induced bone loss. AB - In the present study, we have attempted to evaluate the pharmacological actions of three major phenolic antidiarrheic ingredients, including 2-methoxyphenol (2MP), 2-methoxy-4-methylphenol (2M4MP) and 2-methoxy-4-ethyphenol (2M4EP), on the functionality and integrity of bone by in vitro and in vivo experimental techniques. Intermittent oral administration of 2M4MP and 2M4EP, but not 2MP, significantly prevented reductions of bone mineral density in total femur, distal femur and tibia, in addition to alterations of several osteoclastic parameters on histomorphometric analysis, when determined 28 days after ovariectomy in mice. All three phenolic ingredients examined significantly inhibited the developmental increase in the number of multinucleated cells positive to tartrate-resistant acid phosphatase staining in cultured mouse osteoclasts differentiated from bone marrow precursors in the presence of both macrophage-colony stimulating factor and receptor activator of nuclear factor-kappaB ligand, which occurred in a concentration-dependent manner at a concentration range of 1 microM-1mM without inducing cell death. Moreover, both 2M4MP and 2M4EP at 1mM not only prevented the cell death induced by 0.5mM H2O2 in cultured rat calvarial osteoblasts, but also suppressed the generation of intracellular reactive oxygen species in osteoblasts exposed to H2O2, with a radical scavenging action as revealed by electron spin resonance analysis. These results suggest that particular phenolic antidiarrheic ingredients may prevent ovariectomy-induced bone loss through a mechanism related to the inhibition of osteoclastogenesis in association with an anti-oxidative property in osteoblasts. PMID- 17078934 TI - Divergent effects of the MEKK-1/JNK pathway on NB2a/d1 differentiation: some activity is required for outgrowth and stabilization of neurites but overactivation inhibits both phenomena. AB - c-Jun N-terminal kinase (JNK), along with its upstream activator MEKK-1, is typically thought of as a stress-activated kinase that mediates apoptosis. However, additional studies indicate that the MEKK-1/JNK pathway mediates critical aspects of neuronal survival and differentiation. Herein, we demonstrate that transfection of differentiated NB2a/d1 cells with a construct expression constitutively activated (ca) MEKK-1 increases levels of phospho-dependent neurofilament (NF) immunoreactivity within perikarya, while expression of a dominant-negative (dn) form of MEKK-1 decreases it. Steady-state levels of perikaryal phospho-NF immunoreactivity are reduced and the increase resulting from expression of caMEKK-1 is prevented, by the JNK inhibitor SP600125, suggesting that JNK is a major downstream effector of MEKK-1 on NF phosphorylation. Unexpectedly, both caMEKK-1 and dnMEKK-1 inhibited neuritogenesis as well as translocation of NFs into newly elaborated neurites. The JNK inhibitor SP600125 also inhibited NF transport in a dose-dependent manner. caMEKK-1 also prevented the increase in NF transport otherwise mediated by MAP kinase. Finally, both caMEKK-1 and dnMEKK-1 prevented initial neuritogenesis. These findings indicate that the MEKK-1/JNK pathway regulates critical aspects of initial outgrowth, and subsequent stabilization of axonal neurites. PMID- 17078933 TI - EGCG mitigates neurotoxicity mediated by HIV-1 proteins gp120 and Tat in the presence of IFN-gamma: role of JAK/STAT1 signaling and implications for HIV associated dementia. AB - Human immunodeficiency virus (HIV)-1 infection of the central nervous system occurs in the vast majority of HIV-infected patients. HIV-associated dementia (HAD) represents the most severe form of HIV-related neuropsychiatric impairment and is associated with neuropathology involving HIV proteins and activation of proinflammatory cytokine circuits. Interferon-gamma (IFN-gamma) activates the JAK/STAT1 pathway, a key regulator of inflammatory and apoptotic signaling, and is elevated in HIV-1-infected brains progressing to HAD. Recent reports suggest green tea-derived (-)-epigallocatechin-3-gallate (EGCG) can attenuate neuronal damage mediated by this pathway in conditions such as brain ischemia. In order to investigate the therapeutic potential of EGCG to mitigate the neuronal damage characteristic of HAD, IFN-gamma was evaluated for its ability to enhance well known neurotoxic properties of HIV-1 proteins gp120 and Tat in primary neurons and mice. Indeed, IFN-gamma enhanced the neurotoxicity of gp120 and Tat via increased JAK/STAT signaling. Additionally, primary neurons pretreated with a JAK1 inhibitor, or those derived from STAT1-deficient mice, were largely resistant to the IFN-gamma-enhanced neurotoxicity of gp120 and Tat. Moreover, EGCG treatment of primary neurons from normal mice reduced IFN-gamma-enhanced neurotoxicity of gp120 and Tat by inhibiting JAK/STAT1 pathway activation. EGCG was also found to mitigate the neurotoxic properties of HIV-1 proteins in the presence of IFN-gamma in vivo. Taken together, these data suggest EGCG attenuates the neurotoxicity of IFN-gamma augmented neuronal damage from HIV-1 proteins gp120 and Tat both in vitro and in vivo. Thus EGCG may represent a novel natural copound for the prevention and treatment of HAD. PMID- 17078935 TI - Catalpol increases hippocampal neuroplasticity and up-regulates PKC and BDNF in the aged rats. AB - Rehmannia, a traditional Chinese medical herb, has a long history in age-related disease therapy. Previous work has indicated that catalpol is a main active ingredient performing neuroprotective effect in rehmannia, while the mechanism underlying the effect remains poorly understood. In this study, we attempt to investigate the effect of catalpol on presynaptic proteins and explore a potential mechanism. The hippocampal levels of GAP-43 and synaptophysin in 3 groups of 4 months (young group), 22-24 months (aged group) and catalpol-treated 22-24 months (catalpol-treated group) rats were evaluated by western blotting. Results clearly showed a significant decrease in synaptophysin (46.6%) and GAP-43 (61.4%) levels in the aged group against the young animals and an increase (45.0% and 31.8% respectively) in the catalpol-treated aged rats in comparison with the untreated aged group. In particular, synaptophysin immunoreactivity (OD) in the dentate granule layer of the hippocampus was increased 0.0251 in the catalpol treated group as compared with the aged group. The study also revealed a catalpol associated increase of PKC and BDNF in the hippocampus of the catalpol-treated group in comparison with the aged rats and highly correlated with synaptophysin and GAP-43. Such positive correlations between presynaptic proteins and signaling molecules also existed in the young group. These results suggested that catalpol could increase presynaptic proteins and up-regulate relative signaling molecules in the hippocampus of the aged rats. Consequently, it seemed to indicate that catalpol might ameliorate age-related neuroplasticity loss by "normalizing" presynaptic proteins and their relative signaling pathways in the aged rats. PMID- 17078936 TI - The O-chain structure from the LPS of the endophytic bacterium Burkholderia cepacia strain ASP B 2D. AB - The O-chain polysaccharide of the lipopolysaccharide from the endophytic bacterium Burkholderia cepacia strain was characterized. The structure was studied by means of chemical analysis and 2D NMR spectroscopy and shown to be the following: -->2)-beta-D-Ribf-(1-->6)-alpha-D-Glcp-(1-->. PMID- 17078937 TI - Platelet nitric oxide synthesis in uremia and malnutrition: a role for L-arginine supplementation in vascular protection? AB - L-arginine is the physiological precursor for nitric oxide (NO) synthesis, and availability and transport of L-arginine modulate the rates of NO biosynthesis in circulating blood cells and the vasculature. NO is involved in many vascular functions such as vasodilation and inhibition of platelet aggregation and adhesion. We have established that reduced plasma L-arginine and NO production and increased tumour necrosis factor-alpha (TNF-alpha), fibrinogen, and C reactive protein levels in malnourished uremic patients are associated with increased aggregability of platelets. Our findings may explain the increased cardiovascular mortality in patients with deficient nutritional status, leading to inflammation, oxidative stress, impaired L-arginine-NO signalling, and platelet activation. The aim of this review is to evaluate whether disturbances in the L-arginine-NO signalling pathway in chronic renal failure and atherosclerosis are affected by malnutrition and inflammation. We have included a brief overview of membrane transporters mediating influx of L-arginine and other cationic amino acids, as these transporters are involved in the potential benefits of L-arginine supplementation and platelet function in malnourished uremic patients. PMID- 17078938 TI - Resveratrol affords protection against peroxynitrite-mediated endothelial cell death: A role for intracellular glutathione. AB - Atherosclerosis, the main cause of cardiovascular disease (CD), is a chronic inflammatory condition associated with an overproduction of oxidant species, namely peroxynitrite, which is a powerful oxidant that reacts directly with all biomolecules. Glutathione is an efficient scavenger of peroxynitrite, so, modulation of glutathione synthesis may provide a strategy to selectively protect cells from this oxidant. Here, we investigated the ability of resveratrol, a component of red wine, to prevent peroxynitrite-mediated endothelial cells toxicity and the underlying mechanism. Bovine aortic endothelial cells (BAEC) in primary cultures were treated with authentic peroxynitrite and the cell viability and intracellular glutathione contents were assessed. Our results demonstrate that a long pre-incubation (14 h) of BAEC with resveratrol (1-50 microM) leads to the endothelial cells rescue from injury triggered by authentic peroxynitrite by a mechanism of up-regulation of the intracellular GSH content, for the highest resveratrol concentration tested. Considering the importance of GSH in regulation of cell life, this capacity of resveratrol provides a new mechanism for its cardioprotective effects and may contribute to the development of novel therapeutic strategies. PMID- 17078939 TI - Thermotropic phase behaviour of lipid bilayers containing carotenoid pigment canthaxanthin: a differential scanning calorimetry study. AB - In this study we address the problem of the effect of canthaxanthin on the thermotropic properties of lipid membranes formed with lipids which differ in the thickness of their hydrophobic core, size of polar heads or presence of the ester carbonyl group. For all the lipids a decrease in main transition enthalpy has been observed, indicating that canthaxanthin alters the membrane properties in its gel phase. The strongest influence of canthaxanthin on main phase transition and pretransition has been observed for the lipid having the thinnest hydrophobic region. Component analysis indicates a distinct cooperativity change, which most probably colligates with the formation of new thermotropic phases. The effect of canthaxanthin has been almost negligible in the case of phosphatidylethanolamines. The absence of the ester carbonyl group results in different thermotropic behavior, especially for low canthaxanthin concentrations. The effect of canthaxanthin is explained in terms of its organization within the membrane. PMID- 17078940 TI - Validation of use of annular once-punched filter paper bloodspot samples for repeat lead testing. AB - BACKGROUND: Lack of a second sample often precludes the ability to perform repeat lead measurements on filter paper bloodspot samples. We investigated whether annular specimens remaining from once-punched filter paper bloodspot specimens could provide accurate Pb measurements when measurements were scaled for the remainder area relative to the original punch area. METHODS: 50 microl bloodspot specimens were prepared using Pb-spiked EDTA whole blood. After removal of 6 mm punches, bitmap images of the remainder specimens were obtained using a scanner. Image analysis was used to determine the bloodspot area of the remainder sample relative to the area of the original punch. Measurement of Pb for punches and for remainder specimens was performed by ICP-MS. RESULTS: Area-corrected Pb measurements for remainder samples were significantly higher than for the punches, by an average factor of 1.52+/-0.12 (p<0.05, n=28). The difference was due to a discontinuity (an increase) in the per-area Pb at the bloodspot perimeter. Area-corrected results for annular specimens that excluded the perimeter were identical to those of the punch. CONCLUSION: Area-corrected Pb measurement using annular once-punched bloodspot remainder specimens can accurately reproduce lead measurements obtained from the original punch when the bloodspot perimeter area is excluded. PMID- 17078941 TI - A computational model of fractionated conflict-control mechanisms in task switching. AB - A feature of human cognition is the ability to monitor and adjust one's own behavior under changing circumstances. A dynamic balance between controlled and rapid responding is needed to adapt to a fluctuating environment. We suggest that cognitive control may include, among other things, two distinct processes. Incongruent stimuli may drive top-down facilitation of task-relevant responses to bias performance toward exploitation vs. exploration. Task or response switches may generally slow responses to bias toward accuracy vs. speed and exploration vs. exploitation. Behavioral results from a task switching study demonstrate these two distinct processes as revealed by higher-order sequential effects. A computational model implements the two conflict-control mechanisms, which allow it to capture many complex and novel sequential effects. Lesion studies with the model demonstrate that the model is unable to capture these effects without the conflict-control loops and show how each monitoring component modulates cognitive control. The results suggest numerous testable predictions regarding the neural substrates of cognitive control. PMID- 17078942 TI - Wavelet time-frequency analysis and least squares support vector machines for the identification of voice disorders. AB - This work describes a novel algorithm to identify laryngeal pathologies, by the digital analysis of the voice. It is based on Daubechies' discrete wavelet transform (DWT-db), linear prediction coefficients (LPC), and least squares support vector machines (LS-SVM). Wavelets with different support-sizes and three LS-SVM kernels are compared. Particularly, the proposed approach, implemented with modest computer requirements, leads to an adequate larynx pathology classifier to identify nodules in vocal folds. It presents over 90% of classification accuracy and has a low order of computational complexity in relation to the speech signal's length. PMID- 17078943 TI - The sea urchin histone gene complement. AB - The only eukaryotic mRNAs that are not polyadenylated are the replication dependent histone mRNAs in metazoans. The sea urchin genome contains two sets of histone genes that encode non-polyadenylated mRNAs. One of these sets is a tandemly repeated gene cluster with a 5.6-kb repeat unit containing one copy of each of the five alpha-histone genes and is present as a single large cluster which spans over 1 Mb. There is a second set of genes, consisting of 39 genes, containing two histone H1 genes, 34 genes encoding core histone proteins (H2a, H2b, H3 and H4) and three genes expressed only in the testis. Unlike vertebrates where these genes are clustered, the sea urchin late histone genes, expressed in embryos, larvae and adults, are dispersed throughout the genome. There are also genes encoding polyadenylated histone mRNAs, which encode histone variants, including all variants found in other metazoans, as well as a unique set of five cleavage stage histone proteins expressed in oocytes. The cleavage stage histone H1 is the orthologue of an oocyte-specific histone H1 protein found in vertebrates. PMID- 17078944 TI - The genomic repertoire for cell cycle control and DNA metabolism in S. purpuratus. AB - A search of the Strongylocentrotus purpuratus genome for genes associated with cell cycle control and DNA metabolism shows that the known repertoire of these genes is conserved in the sea urchin, although with fewer family members represented than in vertebrates, and with some cases of echinoderm-specific gene diversifications. For example, while homologues of the known cyclins are mostly encoded by single genes in S. purpuratus (unlike vertebrates, which have multiple isoforms), there are additional genes encoding novel cyclins of the B and K/L types. Almost all known cyclin-dependent kinases (CDKs) or CDK-like proteins have an orthologue in S. purpuratus; CDK3 is one exception, whereas CDK4 and 6 are represented by a single homologue, referred to as CDK4. While the complexity of the two families of mitotic kinases, Polo and Aurora, is close to that found in the nematode, the diversity of the NIMA-related kinases (NEK proteins) approaches that of vertebrates. Among the nine NEK proteins found in S. purpuratus, eight could be assigned orthologues in vertebrates, whereas the ninth is unique to sea urchins. Most known DNA replication, DNA repair and mitotic checkpoint genes are also present, as are homologues of the pRB (two) and p53 (one) tumor suppressors. Interestingly, the p21/p27 family of CDK inhibitors is represented by one homologue, whereas the INK4 and ARF families of tumor suppressors appear to be absent, suggesting that these evolved only in vertebrates. Our results suggest that, while the cell cycle control mechanisms known from other animals are generally conserved in sea urchin, parts of the machinery have diversified within the echinoderm lineage. The set of genes uncovered in this analysis of the S. purpuratus genome should enhance future research on cell cycle control and developmental regulation in this model. PMID- 17078945 TI - The proarrhythmic antihistaminic drug terfenadine increases spontaneous calcium release in human atrial myocytes. AB - Spontaneous calcium release from the sarcoplasmic reticulum in cardiac myocytes plays a central role in cardiac arrhythmogenesis. Compounds intended for therapeutical use that interfere with intracellular calcium handling may therefore have an undesired proarrhythmic potential. Here we have used isolated human atrial myocytes to compare the effect of the proarrhythmic antihistaminic drug terfenadine with the non-proarrhythmic antihistaminic drugs fexofenadine and rupatadine on intracellular calcium homeostasis. Perforated patch-clamp technique was used to measure ionic currents and to detect spontaneous calcium release from the sarcoplasmic reticulum. Our results show that the compound terfenadine, with known arrhythmogenic effects, inhibits L-type calcium current (I(Ca)) with an IC(50) of 185 nM when cells are stimulated at 1.0 Hz. The inhibitory effect of 0.3 muM terfenadine increased from 19+/-4% at stimulation frequency of 0.2 Hz to 63+/-6% at 2.0 Hz. Moreover, terfenadine also increased spontaneous calcium release from the sarcoplasmic reticulum. At a concentration of 1 muM, terfenadine significantly increased the spontaneous Na-Ca exchange current (I(NCX)) frequency from 0.48+/-0.25 to 1.93+/-0.67 s(-1). In contrast, fexofenadine and rupatadine did not change I(Ca) or the frequency of spontaneous I(NCX). We conclude that the proarrhythmic antihistaminic drug terfenadine alters intracellular calcium handling in isolated human atrial myocytes. This experimental model may be suitable to screen for potential arrhythmogenic side-effects of compounds intended for therapeutical use. PMID- 17078946 TI - Effects of purified endogenous inhibitor of the Na+/Ca2+ exchanger on ouabain induced arrhythmias in the atria and ventricle strips of guinea pig. AB - Previous studies demonstrated that the purified endogenous inhibitor (NCX(IF)) of the cardiac Na(+)/Ca(2+) exchanger (NCX1) has the capacity to modulate cardiac muscle contractility. Here, we tested the effects of purified NCX(IF) on arrhythmias induced by ouabain in the atria and ventricle strips of guinea pig. For the sake of comparison NCX(IF) was compared to lidocaine and KB-R7943. In the ventricle strip, NCX(IF) ( approximately 10 U/ml) results in rapid, complete and stable inhibition of ouabain-induced arrhythmias (the inhibition of arrhythmia is not followed by revival of irregular contractions). Under similar experimental conditions the atria strips require somewhat higher doses of NCX(IF) (25-50 U/ml) for complete suppression of arrhythmia. In the atria strip, NCX(IF) (10-25 U/ml) increases the threshold dose (1 microM) of ouabain for arrhythmia onset 2.2+/-0.5 fold (n=5, p<0.05) as well as prolongs the lag-phase for arrhythmia appearance 4.0+/-0.5-fold (n=5, p<0.01). The lag period for arrhythmia onset was also lengthened (2.0+/-0.4-fold) by NCX(IF) in the ventricle strips (n=6, p<0.002). At low frequency of pacing (1 Hz), all three tested substances, lidocaine, KB-R7943, and NCX(IF) can effectively suppress the ouabain-induced arrhythmia. However, at higher frequency (2 Hz), lidocaine is ineffective in suppressing arrhythmia, whereas KB-R7943 becomes pro-arrhythmic. In contrast to reference drugs, NCX(IF) retains its anti-arrhythmic capacity at high frequencies, either in the atria (n=6, p<0.01) or ventricle (n=5, p<0.05) strips. In conclusion, NCX(IF) results in rapid, effective and stable suppression of arrhythmia both in the atria and ventricle preparations under conditions at which the reference drugs become ineffective. PMID- 17078948 TI - Differentiation assays of bone marrow-derived Multipotent Adult Progenitor Cell (MAPC)-like cells towards neural cells cannot depend on morphology and a limited set of neural markers. AB - There are accumulating studies that report a neurogenic potential of bone marrow derived cells both in vitro as well as in vivo. Most claims of neural "transdifferentiation" have based their conclusions on morphology and neural gene expression. Recently, doubts have been raised about the validity of both outcome parameters since non-neural cells can extend neurites and show aberrant neural gene expression as a response to stress inducing factors. In this study, we compared bone marrow-derived Multipotent Adult Progenitor Cell (MAPC)-like cells and neural stem cells (NSC) in their morphology and neural gene expression profile after neural differentiation using three differentiation protocols. We evaluated the expression of five neuroglial antigens [neurofilament 200 (NF200); beta III tubulin (beta3 tub); tau; Glial Fibrillary Acidic Protein (GFAP); Myelin Basic Protein (MBP) and RIP antigen] using real-time PCR (RT-PCR) and immunocytochemistry (ICC). MAPC-like cells adopted a neural-like morphology in one protocol but a fibroblast-like morphology in the two other protocols. RT-PCR and ICC show that MAPC-like cells already express the neural antigens beta III tubulin and NF200 at baseline, but no upregulation of these genes after exposure to three distinct differentiation protocols was seen. In contrast, NSC adopt neural and glial morphologies with a clear increase in expression of all neuroglial genes in all differentiation protocols used. In conclusion, our data demonstrate that neural-like morphology and expression of a limited set of neural marker genes by MAPC-like cells after differentiation are not absolute proof of neural transdifferentiation because MAPC-like cells only partially meet the criteria which are fulfilled by NSC after neural differentiation. PMID- 17078947 TI - Discrete functional elements required for initiation activity of the Chinese hamster dihydrofolate reductase origin beta at ectopic chromosomal sites. AB - The Chinese hamster dihydrofolate reductase (DHFR) DNA replication initiation region, the 5.8 kb ori-beta, can function as a DNA replicator at random ectopic chromosomal sites in hamster cells. We report a detailed genetic analysis of the DiNucleotide Repeat (DNR) element, one of several sequence elements necessary for ectopic ori-beta activity. Deletions within ori-beta identified a 132 bp core region within the DNR element, consisting mainly of dinucleotide repeats, and a downstream region that are required for ori-beta initiation activity at non specific ectopic sites in hamster cells. Replacement of the DNR element with Xenopus or mouse transcriptional elements from rDNA genes restored full levels of initiation activity, but replacement with a nucleosome positioning element or a viral intron sequence did not. The requirement for the DNR element and three other ori-beta sequence elements was conserved when ori-beta activity was tested at either random sites or at a single specific ectopic chromosomal site in human cells. These results confirm the importance of specific cis-acting elements in directing the initiation of DNA replication in mammalian cells, and provide new evidence that transcriptional elements can functionally substitute for one of these elements in ori-beta. PMID- 17078949 TI - A model of tenascin-X integration within the collagenous network. AB - Tenascin-X is an extracellular matrix protein whose absence leads to an Ehlers Danlos syndrome in humans, characterized mainly by disorganisation of collagen and elastic fibril networks. After producing recombinant full-length tenascin-X in mammalian cells, we find that this protein assembled into disulfide-linked oligomers. Trimers were the predominant form observed using rotary shadowing. By solid phase interaction studies, we demonstrate that tenascin-X interacts with types I, III and V fibrillar collagen molecules when they are in native conformation. The use of tenascin-X variants with large regions deleted indicated that both epidermal growth factor repeats and the fibrinogen-like domain are involved in this interaction. Moreover, we demonstrate that tenascin-X binds to the fibril-associated types XII and XIV collagens. We thus suggest that tenascin X, via trimerization and multiple interactions with components of collagenous fibrils, plays a crucial role in the organisation of extracellular matrices. PMID- 17078950 TI - Mass spectrometry studies of demetallation of haemin by recombinant horse L chain apoferritin and its mutant (E 53,56,57,60 Q). AB - An essential difference between eukaryotic ferritins and bacterioferritins is that the latter contain naturally, in vivo haem as Fe-protoporphyrin IX. This haem is located in a hydrophobic pocket along the 2-fold symmetry axes and is liganded by two Met 52. However, in in vivo studies, a cofactor has been isolated in horse spleen apoferritin similar to protoporphyrin IX; in in vitro experiments, it has been shown that horse spleen apoferritin is able to interact with haem. Studies of haemin (Fe(III)-PPIX) incorporation into horse spleen apoferritin have been carried out, which show that the metal free porphyrin is found in a corresponding pocket to haem in bacterioferritins [Precigoux, G., Yariv, J., Gallois, B., Dautant, A., Courseille, C. and Langlois, d'Estaintot B. (1994) A crystallographic study of haem binding to ferritin. Acta Cryst. D 50, 739-743]. A mechanism of demetallation of haemin by L-chain apoferritin was proposed [Crichton, R.R., Soruco, J.A., Roland, F., Michaux, M.A., Gallois, B., Precigoux, G., Mahy, J.P. and Mansuy. (1997) Remarkable ability of horse spleen apoferritin to demetallate hemin and to metallate protoporphyrin IX as a function of pH. J. P. Biochem. 36, 49, 15049-15054]: this involved four Glu residues (53,56,57,60) situated at the entrance of the hydrophobic pocket and appeared to be favoured by acidic conditions. To verify this mechanism, we have mutated these four Glu to Gln and examined demetallation in both acidic and basic conditions. In this paper, we report the mass spectrometry studies of L-chain apoferritin and its mutant incubated with haemin and analysed after different times of incubation: 15 days, 2 months, 6 months, 9 months and 12 months. These studies show that the recombinant L-chain apoferritin and its mutant are able to demetallate haemin to give a hydroxyethyl protoporphyrin IX derivative in a dimeric form [Macieira, S., Martins, B. M. and Huber, R. (2003) Oxygen-dependent coproporphyrinogen IX oxidase from Escherichia coli: one-step purification and biochemical characterization. FEMS. Microbiology Letters 226, 31-37]. PMID- 17078951 TI - PKA modulates GSK-3beta- and cdk5-catalyzed phosphorylation of tau in site- and kinase-specific manners. AB - Phosphorylation of tau protein is regulated by several kinases, especially glycogen synthase kinase 3beta (GSK-3beta), cyclin-dependent protein kinase 5 (cdk5) and cAMP-dependent protein kinase (PKA). Phosphorylation of tau by PKA primes it for phosphorylation by GSK-3beta, but the site-specific modulation of GSK-3beta-catalyzed tau phosphorylation by the prephosphorylation has not been well investigated. Here, we found that prephosphorylation by PKA promotes GSK 3beta-catalyzed tau phosphorylation at Thr181, Ser199, Ser202, Thr205, Thr217, Thr231, Ser396 and Ser422, but inhibits its phosphorylation at Thr212 and Ser404. In contrast, the prephosphorylation had no significant effect on its subsequent phosphorylation by cdk5 at Thr181, Ser199, Thr205, Thr231 and Ser422; inhibited it at Ser202, Thr212, Thr217 and Ser404; and slightly promoted it at Ser396. These studies reveal the nature of the inter-regulation of tau phosphorylation by the three major tau kinases. PMID- 17078952 TI - Changes of gonadal CB1 cannabinoid receptor mRNA in the gilthead seabream, Sparus aurata, during sex reversal. AB - Two cannabinoid receptor-like genes (CB1-like), named CB1A and CB1B, have been isolated in teleost fish, specifically in the puffer fish, Fugu rubripes. However, information on the physiological roles, such as the control of reproduction and development in fish is still scarce. Therefore, the aim of the present study was to investigate the presence of CB1-like mRNA in the gonads of a marine teleost species, the gilthead seabream, Sparus aurata, a hermaphrodite species in which the gonadal tissues first develop as testes, and then as functional ovary. We isolated an 890 bp fragment (GenBank accession number ); that corresponded to the open reading frame of the teleost CB1 receptor gene, encoding for the central portion of the protein, which was aligned with the other bony fish sequence. Using "in situ" hybridization, CB1-like mRNA was localized in both mature and sex-reversing gonads, and relative changes in CB1-like expression levels were detected through semi-quantitative RT-PCR. In the mature testis and in the testicular part of the sex-reversing gonad, CB1 expression levels were found to be much higher compared to the ovarian portion. This suggests that the CB1 signaling is likely involved in the process of testicular regression of the S. aurata, but its actual role has yet to be determined. PMID- 17078954 TI - A review of traumatic airway injuries: potential implications for airway assessment and management. AB - BACKGROUND: Obtaining a patent airway can be difficult in patients with traumatic airway injuries (TAI). There is a paucity of data available about the incidence of airway compromise and techniques used in these patients. METHODS: Charts review of all patients with TAI treated in a Regional Trauma Center from July 1989 to June 2005. RESULTS: One hundred and four patients were identified as TAI in the study period (incidence of 0.4% for blunt and 4.5% for penetrating trauma). Sixty-eighty patients were victims of penetrating trauma (ISS: 24+/-10; mortality: 16%). Thirty-six patients were blunt trauma victims (ISS: 33+/-16; mortality: 36%). Overall, 65% of the patients received a definitive airway (DA) in the pre-hospital setting or at the initial hospital assessment. Alternative techniques for obtaining DA including wound tracheal tube, surgical airway and intubation under fiberoptic bronchoscopy were used in 30% of the patients. Among 24 deaths, 10 were considered primarily due to the airway injury. Twelve patients presented with thoracic TAI with nine deaths in this subgroup. CONCLUSIONS: Overall, the incidence of TAI is low. Blunt trauma TAI is less common, and these patients have a different clinical presentation, higher ISS and mortality than the penetrating TAI group. Early assessment of airways is crucial and DA was required in 2/3 of the patients with TAI. Lower airway injuries have higher mortality than upper airway injuries. Even though most patients died as a result of other injuries, causative factors of death included difficulty in obtaining DA and ventilation/oxygenation problems. PMID- 17078953 TI - Post-castration retention of reproductive behavior and olfactory preferences in male Siberian hamsters: role of prior experience. AB - Reproductive behavior of virtually all adult male rodents is dependent on concurrent availability of gonadal steroids. The ejaculatory reflex is incompatible with long-term absence of testicular steroids and typically disappears within 3 weeks after castration. Male Siberian hamsters are an exception to this rule; mating culminating in the ejaculatory reflex occurs as many as 6 months after castration (persistent copulation). The emergence of persistent copulation many weeks after gonadectomy is here shown not to require repeated post-castration sexual experience. Preoperative sexual experience, on the other hand, significantly increases the percent of males that copulate after gonadectomy, but is not required for the emergence of this trait in 25% of males. Castration prior to puberty prevents persistent copulation in all individuals in adulthood. Persistent copulators, unlike males that cease mating activity after castration, prefer the odors of estrous over non-estrous females when tested 4 months after castration and 7 weeks after the last mating test. Neural circuits of persistent copulators retain the ability to mediate male sex behavior and preferences for female odors in the complete absence of gonadal steroids; they are influenced by preoperative sexual experience and organizational effects of gonadal hormones at the time of puberty. PMID- 17078955 TI - Impact of clinical decision rules on clinical care of traumatic injuries to the foot and ankle, knee, cervical spine, and head. AB - INTRODUCTION: Traumatic injuries to the ankle/foot, knee, cervical spine, and head are very commonly seen in emergency and accident departments around the world. There has been much interest in the development of clinical decision rules to help guide the investigations of these patients in a standardised and cost effective manner. METHODS: In this article we reviewed the impact of the Ottawa ankle rules, Ottawa knee rules, Canadian C-spine rule and the Canadian CT head rule. RESULTS: The studies conducted have confirmed that the use of well developed clinical decision rules results in less radiography, less time spent in the emergency department and does not decrease patient satisfaction or result in misdiagnosis. CONCLUSIONS: Emergency physicians around the world should adopt the use of clinical decision rules for ankle/foot, knee, cervical spine and minor head injuries. With relatively simple implementation strategies, care can be standardized and costs reduced while providing excellent clinical care. PMID- 17078956 TI - Reverse LISS plating for proximal segmental femoral fractures in the polytrauma patient: a case report. PMID- 17078957 TI - Relationships between income, subjective health and caregiver burden in caregivers of people with dementia in group living care: a cross-sectional community-based study. AB - BACKGROUND: Family caregivers of relatives with dementia report higher level of psychological distress than other caregivers and report their self-related health as poorer than that of comparison groups. AIMS AND OBJECTIVES: The purpose of the study was to examine characteristics of family caregivers and to assess whether income, subjective health, age and relationship were associated with the burden of care they experienced. SETTING: Group living units in southern Sweden. PARTICIPANTS: Fifty caregivers who served as informal caregivers of relatives with dementia in group living care. DESIGN: Interviews regarding economic and social conditions and well-evaluated scales for health and caregiver burden (CB) were used. RESULTS: The majority of the family caregivers were adult children, and twice as many were female than were males. The investigation showed that total burden, strain and disappointment, adjusted for health and age, were related to income. Disappointment showed a relation to subjective health. The adult children showed a significantly higher degree of total burden, irrespective of age, compared to other family caregivers. Low income was associated with a higher degree of burden among adult children. However, elderly participants experienced less of burden than younger ones. CONCLUSION: Our findings indicate that caregivers with low health profile and low income, especially adult children, are associated with higher CB. RELEVANCE TO CLINICAL PRACTICE: People with coexisting risk factors (low income, low perceived health) are the ones who may benefit most from health-oriented interventions. PMID- 17078958 TI - Relationship between bone tissue strain and lattice strain of HAp crystals in bovine cortical bone under tensile loading. AB - Cortical bone is a composite material composed of hydroxyapatite (HAp) and collagen. As HAp is a crystalline structure, an X-ray diffraction method is available to measure the strain of HAp crystals. However, HAp crystals in bone tissue have been known to have the low degree of crystallization. Authors have proposed an X-ray diffraction method to measure the lattice strain of HAp crystals from the diffusive intensity profile due to low crystallinity. The precision of strain measurement was greatly improved by this method. In order to confirm the possibility of estimating the bone tissue strain with measurements of the strain of HAp crystals, this work investigates the relationship between bone tissue strain on a macroscopic scale and the lattice strain of HAp crystals on a microscopic scale. The X-ray diffraction experiments were performed under tensile loading. Strip bone specimens of 40x6x0.8mm in size were cut from the cortical region of a shaft of bovine femur. A stepwise tensile load was applied in the longitudinal direction of the specimen. By detecting the diffracted X-ray beam transmitted through the specimen, the lattice strain was directly measured in the loading direction. As a result, the lattice strain of HAp crystals showed lower value than the bone tissue strain measured by a strain gage. The bone tissue strain was described with the mean lattice strain of the HAp crystals and the elastic modulus. PMID- 17078959 TI - Modification of poly(dimethylsiloxane) microfluidic channels with silica nanoparticles based on layer-by-layer assembly technique. AB - A hydrophilic poly(dimethylsiloxane) (PDMS) microchip with stable electroosmotic flow (EOF) was prepared by a simple and reproducible coating procedure with silica nanoparticles. The microchannel wall of PDMS chip was coated with a layer of poly(diallyldimethylammonium chloride) (PDDA) and then collected silica nanoparticles. The assembly was followed by contact angle, charge-coupled device (CCD) imaging, electroosmotic flow (EOF) measurements and electrophoretic separation experiments. Contact angle measurements revealed the coated surface was hydrophilic; the water contact angle for coated chips was 64 degrees compared with a water contact angle for native PDMS chips of 113 degrees . CCD images indicated a substantially more hydrophilic microchannel than native PDMS. We carried out a comparison and concluded that the EOF values on the coated PDMS chip were close to those values on the glass chip above pH 7.0. The coated channel had an excellent stability and reproducibility, RSD of EOF values (n=6) on native and coated PDMS microchip was 1.58 and 0.57%, respectively. Separation of dopamine and epinephrine was performed on the coated chip generated 1.40 x 10(5), 1.39 x 10(5) theoretical plates/m compared with the native PDMS chip of 0.79 x 10(5), 0.88 x 10(5), high resolution of 1.7 was achieved with a channel of 3.60 cm length. PMID- 17078960 TI - Short-cut method for the correction of light attenuation influences in the experimental data obtained from confocal laser scanning microscopy. AB - Based on Lambert-Beer law and the light attenuation model, a new method will be introduced in this work in order to eliminate the disturbances caused by signal attenuation in the experimental data measured by confocal laser scanning microscopy (CLSM). This new method considers the attenuation effects which depend on concentration of fluorophore-labelled protein as well as attenuation effects which are independent from protein concentration. Furthermore, no solvent additive is required in order to match the refraction index of solvent to bead material. The determination of correction factors is, thus, easily done using the currently investigated chromatographic phase system, so that the validity of the correction factors in the current system can be guaranteed. The introduced correction method has been applied for the investigation of intraparticle protein distribution inside an HIC (hydrophobic interaction chromatography) particle. PMID- 17078961 TI - Estimation of the Hamaker constants by sedimentation field-flow fractionation. AB - van der Waals forces are one of several forces that control the adhesion between two materials. These forces are important to quantify in adhesion studies because they are always present and are always attractive. The major problem in calculating the van der Waals interaction between colloidal particles is that of evaluating the Hamaker constant. Hence, an accurately determined Hamaker constant for a given material is needed when interfacial phenomena such as adhesion are discussed in terms of the total potential energy between a particle and a substrate. In this paper, a new simple and accurate methodology for the estimation of the Hamaker constant is introduced. The results are in good agreement with those values found in literature. PMID- 17078962 TI - Fast gradient elution reversed-phase high-performance liquid chromatography with diode-array detection as a high-throughput screening method for drugs of abuse. I. Chromatographic conditions. AB - A new approach for the high-throughput screening of biological samples to detect the presence of regulated intoxicants has been developed by modifying a conventional gradient elution high-performance liquid chromatograph (HPLC). The goal of this work was to improve the speed of gradient elution screening methods over current approaches by optimizing the operational parameters of both the column and the instrument without compromising the reproducibility of the retention times, which is the basis for the identification of intoxicant compounds. Most importantly, the novel instrument configuration substantially reduces the time needed to re-equilibrate the column between consecutive gradient runs, thereby reducing the total time for each analysis. The total analysis time for each gradient elution run is only 2.80 min, including 0.30 min for column re equilibration between analyses. Retention times of standard calibration solutes are reproducible to better than 0.002 min in consecutive runs. A corrected retention index was adopted to account for day-to-day and column-to-column variations in retention time. For a set of forty-seven target compounds, the discriminating power and mean list length were found to be 0.95 and 3.26, respectively. In comparison to previous work with similar numbers of target compounds, the current approach provides an order of magnitude improvement in analysis time, and a four-fold decrease in mean list length. PMID- 17078963 TI - Expression of androgen, oestrogen alpha and beta, and progesterone receptors in the canine prostate: differences between normal, inflamed, hyperplastic and neoplastic glands. AB - The expression of receptor for androgen (AR), oestrogen alpha and beta (ERalpha and ERbeta) and progesterone (PR) was examined immunohistochemically in canine prostate specimens (normal, hyperplastic, inflamed [prostatitis] or neoplastic). AR immunolabelling was seen in 100% of epithelial cells of normal and hyperplastic tissue, the corresponding figures for inflamed and carcinomatous tissue being 74% and 65%, respectively. ERalpha labelling was seen in 85% of epithelial cells in normal prostate glands, the corresponding figures for hyperplastic, inflamed and neoplastic glands being 35%, 22% and 12%, respectively. ERbeta labelling was seen in 85% of epithelial cells of normal glands and in about 70% of such cells in glands showing pathological changes. On the other hand, PR expression (weak) in normal glands was observed in fewer epithelial cells (44%) than in hyperplastic (70%), inflamed (62%) or neoplastic (64%) glands. PMID- 17078964 TI - Selection of a novel gp41-specific HIV-1 neutralizing human antibody by competitive antigen panning. AB - The HIV envelope glycoprotein (Env) is composed of two non-covalently associated subunits: gp120 and gp41. Panning of phage-displayed antibody libraries against Env-based antigens has resulted mostly in selection of anti-gp120 antibodies. Native gp41 in the absence of gp120 is unstable. The use of gp41 fragments as antigens has resulted in selection of antibodies with only relatively modest neutralizing activity. To enhance selection of antibodies specific for gp41 in the context of the whole Env we developed a methodology termed competitive antigen panning (CAP). Using CAP, we identified a novel gp41-specific human monoclonal antibody (hmAb), m48, from an immune library derived from long-term nonprogressors with high titers of broadly cross-reactive neutralizing antibodies (bcnAbs). Selection of m48 was only successful using CAP and not through the conventional pre-incubation methodology. In assays based on spreading infection in peripheral blood mononuclear cells (PBMCs) m48 neutralized a panel of HIV-1 primary isolates from different clades more potently than the well-characterized broadly cross-reactive HIV-1-neutralizing antibodies IgG1 4E10 and Fab Z13. These results may have implications for the selection of novel gp41-specific bcnAbs and other antibodies, and for the development of HIV-1 inhibitors and vaccine immunogens. PMID- 17078965 TI - Upregulation of two actin genes and redistribution of actin during diapause and cold stress in the northern house mosquito, Culex pipiens. AB - Two actin genes cloned from Culex pipiens L. are upregulated during adult diapause. Though actins 1 and 2 were expressed throughout diapause, both genes were most highly expressed early in diapause. These changes in gene expression were accompanied by a conspicuous redistribution of polymerized actin that was most pronounced in the midguts of diapausing mosquitoes that were exposed to low temperature. In nondiapausing mosquitoes reared at 25 degrees C and in diapausing mosquitoes reared at 18 degrees C, polymerized actin was clustered at high concentrations at the intersections of the muscle fibers that form the midgut musculature. When adults 7-10 days post-eclosion were exposed to low temperature (-5 degrees C for 12 h), the polymerized actin was evenly distributed along the muscle fibers in both nondiapausing and diapausing mosquitoes. Exposure of older adults (1 month post-eclosion) to low temperature (-5 degrees C for 12 h) elicited an even greater distribution of polymerized actin, an effect that was especially pronounced in diapausing mosquitoes. These changes in gene expression and actin distribution suggest a role for actins in enhancing survival of diapausing adults during the low temperatures of winter by fortification of the cytoskeleton. PMID- 17078966 TI - Nutritional balancing in fruit flies: performance of wild adult females of Anastrepha obliqua (Diptera: Tephritidae) fed on single-food or food-pair treatments. AB - The aim of the present study was to investigate the effects of different artificial diets, in which sucrose and brewer's yeast were offered separately (food-pair treatment) or mixed (single-food treatment), on the regulation of nutrient ingestion and the performance of wild adult females of Anastrepha obliqua. The amount of diet consumed by females of A. obliqua increased as the concentration of yeast in the diet increased, revealing a phagostimulant effect of yeast. The amount of sucrose ingested was constant in all groups, showing that these flies regulate the ingestion of sucrose. Longevity and production of eggs/female were similar in both groups. Our results suggest that A. obliqua females regulate the ingestion of nutrients to obtain an adequate performance. Dietary self-selective behavior was more efficient for the females of A. obliqua than feeding on a single-food, because the self-selecting flies ingested less food and had a similar performance as the ones fed on the single-food treatment. PMID- 17078967 TI - "Hiding out" from chronic ischemia with help from the mitochondria? PMID- 17078968 TI - Janus model of the Na,K-ATPase beta-subunit transmembrane domain: distinct faces mediate alpha/beta assembly and beta-beta homo-oligomerization. AB - Na,K-ATPase is a hetero-oligomer of alpha and beta-subunits. The Na,K-ATPase beta subunit (Na,K-beta) is involved in both the regulation of ion transport activity, and in cell-cell adhesion. By structure prediction and evolutionary analysis, we identified two distinct faces on the Na,K-beta transmembrane domain (TMD) that could mediate protein-protein interactions: a glycine zipper motif and a conserved heptad repeat. Here, we show that the heptad repeat face is involved in the hetero-oligomeric interaction of Na,K-beta with Na,K-alpha, and the glycine zipper face is involved in the homo-oligomerization of Na,K-beta. Point mutations in the heptad repeat motif reduced Na,K-beta binding to Na,K-alpha, and Na,K ATPase activity. Na,K-beta TMD homo-oligomerized in biological membranes, and mutation of the glycine zipper motif affected oligomerization and cell-cell adhesion. These results provide a structural basis for understanding how Na,K beta links ion transport and cell-cell adhesion. PMID- 17078969 TI - Characteristics affecting expression and solubilization of yeast membrane proteins. AB - Biochemical and structural analysis of membrane proteins often critically depends on the ability to overexpress and solubilize them. To identify properties of eukaryotic membrane proteins that may be predictive of successful overexpression, we analyzed expression levels of the genomic complement of over 1000 predicted membrane proteins in a recently completed Saccharomyces cerevisiae protein expression library. We detected statistically significant positive and negative correlations between high membrane protein expression and protein properties such as size, overall hydrophobicity, number of transmembrane helices, and amino acid composition of transmembrane segments. Although expression levels of membrane and soluble proteins exhibited similar negative correlations with overall hydrophobicity, high-level membrane protein expression was positively correlated with the hydrophobicity of predicted transmembrane segments. To further characterize yeast membrane proteins as potential targets for structure determination, we tested the solubility of 122 of the highest expressed yeast membrane proteins in six commonly used detergents. Almost all the proteins tested could be solubilized using a small number of detergents. Solubility in some detergents depended on protein size, number of transmembrane segments, and hydrophobicity of predicted transmembrane segments. These results suggest that bioinformatic approaches may be capable of identifying membrane proteins that are most amenable to overexpression and detergent solubilization for structural and biochemical analyses. Bioinformatic approaches could also be used in the redesign of proteins that are not intrinsically well-adapted to such studies. PMID- 17078970 TI - Planning genetic studies on primary adult-onset dystonia: sample size estimates based on examination of first-degree relatives. AB - Primary adult-onset dystonia is thought to be partly genetic, but families large enough for a genome wide search are difficult to find. We examined the first degree relatives of 76 primary adult-onset dystonia patients to assess the feasibility of model-free nonparametric methods that allow either screening of candidate loci (case-control design, transmission disequilibrium test [TDT], and sibling-TDT [S-TDT]) or identification of novel genes (affected sib-pair [ASP] method). Among the examined relatives, 1/34 parents, 13/149 siblings and 10/125 offspring were affected by adult-onset dystonia. The predicted sample sizes to detect a gene conferring an Odds ratio of 3.0 were 99 for case-control and TDT methodology, 148 for S-TDT, and 107 to 173 for an ASP study assuming three major loci. Based on our family structure, TDT, S-TDT, and ASP methods would required screening of about 220, 700, and 580 to 939 probands respectively. Analysing subpopulations with different types of dystonia, TDT required fewer probands with cervical/hand dystonia, S-TDT needed fewer probands with cranial dystonia. These sample size estimates suggest that the S-TDT may be feasible, whereas collection of cases for both TDT and ASP approaches would represent a major collaborative challenge. PMID- 17078971 TI - The cortico-diaphragmatic pathway involvement in amyotrophic lateral sclerosis: neurophysiological, respiratory and clinical considerations. AB - Cortico-diaphragmatic pathway was investigated by means of transcranial magnetic stimulation (TMS), in 14 patients affected by definite amyotrophic lateral sclerosis (ALS) without clinical signs of respiratory impairment. Spirometry, gas analysis, and measurement of static inspiratory and expiratory pressures were performed in all patients. Forced vital capacity, forced expiratory volume at the first and second peak expiratory flow, sniff effort from FRC level (SNIP), maximal inspiratory and expiratory pressure at mouth (MIP/MEP), maximal transdiaphragmatic pressure (Pdimx) were considered. TMS was performed, recording by surface electrodes from hemidiaphragm, bilaterally. Latency of cortical and spinal motor-evoked potentials (Cx-MEP/Sp-MEP) and central motor conduction time (CMCT) were measured. None of the patients showed altered spirometry and gas levels. Seven patients showed decreased Pdimx and eight of MEP values. Four patients showed a delayed Sp-MEP. In one patient the Cx-MEP was abolished while the mean values of both Cx-MEP and CMCT were significantly increased (19.2+/-4.1 ms, P<0.0001; 10.8+/-4.8 ms, P<0.0001). Cx-MEP and CMCT did not show significant correlations with any of the respiratory measures. The patients with prolonged Sp MEP, showed longer disease duration, lower Norris score, lower Pdimx and MEP values. In conclusion, cortico-diaphragmatic study is a sensitive measure to reveal subclinical diaphragmatic impairment although not correlated to respiratory measures. PMID- 17078972 TI - The evolution of the genetic code took place in an anaerobic environment. AB - We have compared orthologous proteins from an aerobic organism, Cytophaga hutchinsonii, and from an obligate anaerobe, Bacteroides thetaiotaomicron. This comparison allows us to define the oxyphobic ranks of amino acids, i.e. a scale of the relative sensitivity to oxygen of the amino acid residues. The oxyphobic index (OI), which can be simply obtained from the amino acids' oxyphobic ranks, can be associated to any protein and therefore to the genetic code, if the number of synonymous codons attributed to the amino acids in the code is assumed to be the frequency with which the amino acids appeared in ancestral proteins. Sampling of the OI variable from the proteins of obligate anaerobes and aerobes has established that the OI value of the genetic code is not significantly different from the mean OI value of anaerobe proteins, while it is different from that of aerobe proteins. This observation would seem to suggest that the terminal phases of the evolution of genetic code organization took place in an anaerobic environment. This result is discussed in the framework of hypotheses suggested to explain the timing of the evolutionary appearance of the aerobic metabolism. PMID- 17078974 TI - Environmental impacts of dredging on seagrasses: a review. AB - Main potential impacts on seagrasses from dredging and sand mining include physical removal and/or burial of vegetation and effects of increased turbidity and sedimentation. For seagrasses, the critical threshold for turbidity and sedimentation, as well as the duration that seagrasses can survive periods of high turbidity or excessive sedimentation vary greatly among species. Larger, slow-growing climax species with substantial carbohydrate reserves show greater resilience to such events than smaller opportunistic species, but the latter display much faster post-dredging recovery when water quality conditions return to their original state. A review of 45 case studies worldwide, accounting for a total loss of 21,023 ha of seagrass vegetation due to dredging, is indicative of the scale of the impact of dredging on seagrasses. In recent years, tighter control in the form of strict regulations, proper enforcement and monitoring, and mitigating measures together with proper impact assessment and development of new environmental dredging techniques help to prevent or minimize adverse impacts on seagrasses. Costs of such measures are difficult to estimate, but seem negligible in comparison with costs of seagrass restoration programmes, which are typically small-scale in approach and often have limited success. Copying of dredging criteria used in one geographic area to a dredging operation in another may in some cases lead to exaggerated limitations resulting in unnecessary costs and delays in dredging operations, or in other cases could prove damaging to seagrass ecosystems. Meaningful criteria to limit the extent and turbidity of dredging plumes and their effects will always require site-specific evaluations and should take into account the natural variability of local background turbidity. PMID- 17078973 TI - Tracking the dynamics of pathogen interactions: modeling ecological and immune mediated processes in a two-pathogen single-host system. AB - Traditionally, epidemiological studies have focused on understanding the dynamics of a single pathogen, assuming no interactions with other pathogens. Recently, a large body of work has begun to explore the effects of immune-mediated interactions, arising from cross-immunity and antibody-dependent enhancement, between related pathogen strains. In addition, ecological processes such as a temporary period of convalescence and pathogen-induced mortality have led to the concept of ecological interference between unrelated diseases. There remains, however, the need for a systematic study of both immunological and ecological processes within a single framework. In this paper, we develop a general two pathogen single-host model of pathogen interactions that simultaneously incorporates these mechanisms. We are then able to mechanistically explore how immunoecological processes mediate interactions between diseases for a pool of susceptible individuals. We show that the precise nature of the interaction can induce either competitive or cooperative associations between pathogens. Understanding the dynamic implications of multi-pathogen associations has potentially important public health consequences. Such a framework may be especially helpful in disentangling the effects of partially cross-immunizing infections that affect populations with a pre-disposition towards immunosuppression such as children and the elderly. PMID- 17078975 TI - Occurrence of organochlorine pesticides, polychlorinated biphenyls and polybrominated diphenyl ethers in deep-sea fishes from the Sulu Sea. PMID- 17078976 TI - Surface tension effects on instability in viscoelastic respiratory fluids. AB - This paper establishes the mathematical formalism for the modeling of the mucus layer in the human trachea as a viscoelastic multiphase fluid system with surface tension with a view toward study of instability properties of the air-mucus system aimed at improving the design of new bioaerosol suppressing medication. The effects of surface tension, previously only conjectured and very poorly understood, are clearly established with quantitative relationships. Several very important physiological conclusions are obtained supporting one method of potential treatment and prevention of disease transmission by alteration of the mucus layer properties over other potential methods. PMID- 17078977 TI - Rapid changes in the sensitivity of arcuate nucleus neurons to central ghrelin in relation to feeding status. AB - Ghrelin, the endogenous ligand for the growth hormone secretagogue (GHS) receptor, stimulates feeding and increases body weight. Systemic ghrelin administration induces the immediate-early gene protein product, c-Fos, in the arcuate nucleus of the hypothalamus (ARC) of satiated rats and this increase is potentiated in fasted rats. The aim of this study was to determine whether potentiation was seen in fasted animals after intracerebroventricular (i.c.v) administration of ghrelin and to identify the hypothalamic nuclei activated by this peptide. In addition we investigated if allowing fasted animals to re-feed for 1 h prior to i.c.v. ghrelin injection affected the c-Fos response. Using c Fos immunocytochemistry, we demonstrated that i.c.v. ghrelin activated several hypothalamic nuclei, including the ARC, paraventricular nucleus (PVH) and the lateral hypothalamus (LH). The c-Fos response was greater in fasted animals compared with satiated animals. Fasted rats allowed access to food for 1 h prior to central ghrelin administration showed an attenuated response in the ARC, similar to the response seen in fed animals. However, the response in the LH (including in the orexin neurons) was further potentiated. The latter may reflect a connection between the hypothalamus and regions of the brain responding to the reward value of the meal. PMID- 17078978 TI - Oral behavior from food intake until terminal swallow. AB - We analyzed oral behavior from food intake until terminal swallow for mastication and swallowing under a freely eating condition with a natural food. Measurements, including movement of the mandible and tongue, the size of the gape, different sequences involved in the oral aspect of the swallowing action, and bolus size and movement were carried out in five "freely eating subjects" using videofluorography. During food intake, the tongue moved forwards and backwards to introduce food into the mouth, to compress the food against the hard palate, and to transport food to the occlusal surface of the molar teeth. Most of the food was swallowed in the first swallow, and any residual food was aggregated by the tongue into a bolus and then swallowed in the last swallow. These findings suggest that 1) tongue manipulation plays an important role in recognizing and evaluating the volume of bite taken, 2) the intra-oral compression of food has a role in the recognition of food texture, 3) stage I transport is closely bound to the texture recognition process, 4) humans need at least two swallows, even with one bite of food, when ingesting food freely, and 5) the duration time of the oral stage of swallowing may depend on the bolus volume and be longer for smaller volumes unlike those measured under the command swallow. PMID- 17078979 TI - The role of breakfast and a mid-morning snack on the ability of children to concentrate at school. AB - The effect on the ability of children to attend to their school work, of the size of breakfast and whether a mid-morning snack had been consumed, was considered. Nine year old children were studied for four days. They reported what they had eaten for breakfast and days when they either had or had not eaten a mid-morning snack were contrasted. For an hour in the late morning, while performing individual work, activity sampling was used to establish the time spent on task. Those who had eaten a small breakfast, on average 61 kcal, spent significantly less time attending to their work than those who had eaten larger meals. The adverse effect of a small breakfast was reversed by the consumption of a mid morning snack. PMID- 17078980 TI - Learned contextual cue potentiates eating in rats. AB - Explicit cues associated with food consumption when hunger prevails will enhance eating when they are subsequently presented under conditions of satiety. Here we examined whether contextual conditioned stimuli (CSs) paired with consumption of food pellets while rats were food-deprived would enhance consumption of this food in rats that were not food-deprived. The conditioning context enhanced rats' consumption of the training food, but it did not change their consumption of the familiar, lab chow. These results show that the contextual CSs, like discrete cues, could modulate food consumption in a CS-potentiated eating paradigm. Furthermore, the data suggest that CS-potentiation of eating does not induce a general motivation to eat, akin to hunger, but instead more likely produces a more specific motivational state, akin to craving. PMID- 17078981 TI - Dietary tyrosine benefits cognitive and psychomotor performance during body cooling. AB - Supplemental tyrosine is effective at limiting cold-induced decreases in working memory, presumably by augmenting brain catecholamine levels, since tyrosine is a precursor for catecholamine synthesis. The effectiveness of tyrosine for preventing cold-induced decreases in physical performance has not been examined. This study evaluated the effect of tyrosine supplementation on cognitive, psychomotor, and physical performance following a cold water immersion protocol that lowered body core temperature. Fifteen subjects completed a control trial (CON) in warm (35 degrees C) water and two cold water trials, each spaced a week apart. Subjects ingested an energy bar during each trial; on one cold trial (TYR) the bar contained tyrosine (300 mg/kg body weight), and on the other cold trial (PLB) and on CON the bar contained no tyrosine. Following each water immersion, subjects completed a battery of performance tasks in a cold air (10 degrees C) chamber. Core temperature was lower (p=0.0001) on PLB and TYR (both 35.5+/-0.6 degrees C) than CON (37.1+/-0.3 degrees C). On PLB, performance on a Match-to Sample task decreased 18% (p=0.02) and marksmanship performance decreased 14% (p=0.002), compared to CON, but there was no difference between TYR and CON. Step test performance decreased by 11% (p=0.0001) on both cold trials, compared to CON. These data support previous findings that dietary tyrosine supplementation is effective for mitigating cold-induced cognitive performance such as working memory, even with reduced core temperature, and extends those findings to include the psychomotor task of marksmanship. PMID- 17078982 TI - Molecular cloning and functional expression of a multifunctional triterpene synthase cDNA from a mangrove species Kandelia candel (L.) Druce. AB - Homology based PCRs with degenerate primers designed from the conserved sequences among the known oxidosqualene cylases (OSCs) have resulted in cloning of a triterpene synthase (KcMS) from the young roots of Kandelia candel (L.) Druce (Rhizophoraceae). KcMS consists of a 2286 bp open reading frame, which codes for 761 amino acids. The deduced amino acid sequence showed 79% homology to a lupeol synthase from Ricinus communis suggesting it to be a lupeol synthase of K. candel. KcMS was expressed in a lanosterol synthase deficient yeast with the expression vector pYES2 under the control of GAL1 promoter. GC-MS analysis showed that the transformant accumulated a mixture of lupeol, beta-amyrin and alpha amyrin in a 2:1:1 ratio, indicating that KcMS encodes a multifunctional triterpene synthase, although it showed high sequence homology to a R. communis lupeol synthase. This is the first OSC cloning from mangrove tree species. PMID- 17078983 TI - Carboxylesterase activities toward pesticide esters in crops and weeds. AB - Proteins were extracted from maize, rice, sorghum, soybean, flax and lucerne; the weeds Abutilon theophrasti, Echinochloa crus-galli, Phalaris canariensis, Setaria faberii, Setaria viridis, Sorghum halepense and the model plant Arabidopsis thaliana and assayed for carboxylesterase activity toward a range of xenobiotics. These included the pro-herbicidal esters clodinafop-propargyl, fenoxaprop-ethyl, fenthioprop-ethyl, methyl-2,4-dichlorophenoxyacetic acid (2,4-d-methyl), bromoxynil-octanoate, the herbicide-safener cloquintocet-mexyl and the pyrethroid insecticide permethrin. Highest activities were recorded with alpha-naphthyl acetate and methylumbelliferyl acetate. Esters of p-nitrophenol were also readily hydrolysed, with turnover declining as the chain length of the acyl component increased. Activities determined with model substrates were much higher than those observed with pesticide esters and were of limited value in predicting the relative rates of hydrolysis of the crop protection agents. Substrate preferences with the herbicides were typically 2,4-d-methyl>clodinafop-propargyl>fenthioprop ethyl, fenoxaprop-ethyl and bromoxynil-octanoate. Isoelectric focussing in conjunction with staining for esterase activity using alpha-naphthyl acetate as substrate confirmed the presence of multiple carboxylesterase isoenzymes in each plant, with major qualitative differences observed between species. The presence of serine hydrolases among the resolved isoenzymes was confirmed through their selective inhibition by the organophosphate insecticide paraoxon. Our studies identify potentially exploitable differences between crops and weeds in their ability to bioactivate herbicides by enzymic hydrolysis and also highlight the usefulness of Arabidopsis as a plant model to study xenobiotic biotransformation. PMID- 17078984 TI - Sugar for my honey: carbohydrate partitioning in ectomycorrhizal symbiosis. AB - Simple, readily utilizable carbohydrates, necessary for growth and maintenance of large numbers of microbes are rare in forest soils. Among other types of mutualistic interactions, the formation of ectomycorrhizas, a symbiosis between tree roots and certain soil fungi, is a way to overcome nutrient and carbohydrate limitations typical for many forest ecosystems. Ectomycorrhiza formation is typical for trees in boreal and temperate forests of the northern hemisphere and alpine regions world-wide. The main function of this symbiosis is the exchange of fungus-derived nutrients for plant-derived carbohydrates, enabling the colonization of mineral nutrient-poor environments. In ectomycorrhizal symbiosis up to 1/3 of plant photoassimilates could be transferred toward the fungal partner. The creation of such a strong sink is directly related to the efficiency of fungal hexose uptake at the plant/fungus interface, a modulated fungal carbohydrate metabolism in the ectomycorrhiza, and the export of carbohydrates towards soil growing hyphae. However, not only the fungus but also the plant partner increase its expression of hexose importer genes at the plant/fungus interface. This increase in hexose uptake capacity of plant roots in combination with an increase in photosynthesis may explain how the plant deals with the growing fungal carbohydrate demand in symbiosis and how it can restrict this loss of carbohydrates under certain conditions to avoid fungal parasitism. PMID- 17078985 TI - Arbuscular mycorrhiza and heavy metal tolerance. AB - Arbuscular mycorrhizal fungi (AMF) have repeatedly been demonstrated to alleviate heavy metal stress of plants. The current manuscript summarizes results obtained to date on the colonization of plants by AMF in heavy metal soils, the depositions of heavy metals in plant and fungal structures and the potential to use AMF-plant combinations in phytoremediation, with emphasis on pennycresses (Thlaspi ssp.). The focus of this manuscript is to describe and discuss studies on the expression of genes in plants and fungi under heavy metal stress. The summary is followed by data on differential gene expression in extraradical mycelia (ERM) of in vitro cultured Glomus intraradices Sy167 supplemented with different heavy metals (Cd, Cu or Zn). The expression of several genes encoding proteins potentially involved in heavy metal tolerance varied in their response to different heavy metals. Such proteins included a Zn transporter, a metallothionein, a 90 kD heat shock protein and a glutathione S-transferase (all assignments of protein function are putative). Studies on the expression of the selected genes were also performed with roots of Medicago truncatula grown in either a natural, Zn-rich heavy metal "Breinigerberg" soil or in a non-polluted soil supplemented with 100 microM ZnSO(4). The transcript levels of the genes analyzed were enhanced up to eight fold in roots grown in the heavy metal containing soils. The data obtained demonstrate the heavy metal-dependent expression of different AMF genes in the intra- and extraradical mycelium. The distinct induction of genes coding for proteins possibly involved in the alleviation of damage caused by reactive oxygen species (a 90 kD heat shock protein and a glutathione S-transferase) might indicate that heavy metal-derived oxidative stress is the primary concern of the fungal partner in the symbiosis. PMID- 17078986 TI - Blood concentrations of amino acids, glucose and lactate during experimental swine dysentery. AB - The aim of this study was to examine blood concentrations of amino acids, glucose and lactate in association with experimental swine dysentery. Ten pigs (approximately 23kg) were orally inoculated with Brachyspira hyodysenteriae. Eight animals developed muco-haemorrhagic diarrhoea with impaired general appearance, changes in white blood cell counts and increased levels of the acute phase protein Serum Amyolid A. Blood samples were taken before inoculation, during the incubation period, during clinical signs of dysentery and during recovery. Neither plasma glucose nor lactate concentrations changed during the course of swine dysentery, but the serum concentrations of gluconeogenic non essential amino acids decreased during dysentery. This was mainly due to decreases in alanine, glutamine, serine and tyrosine. Lysine increased during dysentery and at the beginning of the recovery period, and leucine increased during recovery. Glutamine, alanine and tyrosine levels show negative correlations with the numbers of neutrophils and monocytes. In conclusion, swine dysentery altered the blood concentrations of amino acids, but not of glucose or lactate. PMID- 17078987 TI - Diquat induces renal proximal tubule injury in glutathione reductase-deficient mice. AB - Reactive oxygen species (ROS) have been associated with many human diseases, and glutathione (GSH)-dependent processes are pivotal in limiting tissue damage. To test the hypothesis that Gr1(a1Neu) (Neu) mice, which do not express glutathione reductase (GR), would be more susceptible than are wild-type mice to ROS-mediated injury, we studied the effects of diquat, a redox cycling toxicant. Neu mice exhibited modest, dose- and time-dependent elevations in plasma alanine aminotransferase (ALT) activities, 126+/-36 U/l at 2 h after 5 micromol/kg of diquat, but no ALT elevations were observed in diquat-treated C3H/HeN mice for up to 6 h after 50 micromol/kg of diquat. Histology indicated little or no hepatic necrosis in diquat-treated mice of either strain, but substantial renal injury was observed in diquat-treated Neu mice, characterized by brush border sloughing in the proximal tubules by 1 h and tubular necrosis by 2 h after doses of 7.5 micromol/kg. Decreases in renal GSH levels were observed in the Neu mice by 2 h post dose (3.4+/-0.4 vs 0.2+/-0.0 micromol/g tissue at 0 and 50 micromol/kg, respectively), and increases in renal GSSG levels were observed in the Neu mice as early as 0.5 h after 7.5 micromol/kg (105.5+/-44.1 vs 27.9+/-4.8 nmol/g tissue). Blood urea nitrogen levels were elevated by 2 h in Neu mice after doses of 7.5 micromol/kg (Neu vs C3H, 32.8+/-4.1 vs 17.9+/-0.3 mg/dl). Diquat-induced renal injury in the GR-deficient Neu mice offers a useful model for studies of ROS-induced renal necrosis and of the contributions of GR in defense against oxidant-mediated injuries in vivo. PMID- 17078988 TI - Effects of Russell's viper venom fractions on systemic and renal hemodynamics. AB - Changes in systemic and renal hemodynamics induced by Russell's viper venom are well established. The component of the venom responsible for hemodynamic alteration has not been identified. By Sephadex column chromatography five fractions of Russell's viper (Daboia russellii siamensis) venom were isolated. Each venom fraction consisted of phospholipase A2, proteolytic enzyme, phosphomonoesterase, phosphodiesterase, arginine ester hydrolase and hyaluronidase of varying activities. Hemodynamic effects of each venom fraction including mean arterial pressure, cardiac output, systemic and renal vascular resistance, renal blood flow and glomerular filtration rate were studied in five groups of dogs; each group had four dogs. Minimal hemodynamic changes were observed in dogs receiving venom fraction I. Increased renal vascular resistance with diminution of renal blood flow and glomerular filtration rate was observed in dogs receiving venom fractions II, III, IV and V. A markedly increased renal vascular resistance with maximal decrease in renal blood flow and glomerular filtration rate was caused by fraction III of the venom with highest PLA2 and proteolytic enzyme activities. However, renal hemodynamic changes appeared to correlate better with proteolytic enzyme activity than PLA2 activity. The findings suggested the proteolytic enzyme as an important determinant of hemodynamic alteration. Fractional excretion of Na was increased in dogs injected with venom fraction IV, and is presumed to be due to the inhibition of tubular reabsorption of Na by a natriuretic factor in this venom fraction. PMID- 17078989 TI - The liver of woodchucks chronically infected with the woodchuck hepatitis virus contains foci of virus core antigen-negative hepatocytes with both altered and normal morphology. AB - The livers of woodchucks chronically infected with woodchuck hepatitis virus (WHV) contain foci of morphologically altered hepatocytes (FAH) with "basophilic", "amphophilic" and "clear cell" phenotypes, which are possibly pre neoplastic in nature. Interestingly, most fail to express detectable levels of WHV proteins and nucleic acids. We studied sections of WHV-infected liver tissue to determine if all foci of hepatocytes that failed to express detectable levels of WHV, as assessed by immunoperoxidase staining for WHV core antigen, could be classified morphologically as FAH. We found that at least half of the foci of WHV core antigen-negative hepatocytes did not show clear morphological differences in either H&E or PAS (periodic acid Schiff) stained sections from surrounding hepatocytes, and were therefore not designated as FAH. In the second approach, we assayed core antigen-negative foci for the presence of fetuin B, a serum protein produced by normal hepatocytes, but not by neoplastic hepatocytes in hepatocellular carcinomas. Basophilic and amphophilic FAH had reduced levels of fetuin B compared to hepatocytes present in the surrounding liver; fetuin B staining was detected in clear cell FAH but the level could not be accurately assessed because of the displacement of fetuin B to the cell periphery by accumulated glycogen. The foci of morphologically normal WHV core antigen negative hepatocytes had similar levels of fetuin B to that of the surrounding hepatocytes. The co-existence of at least four types of WHV core antigen-negative foci, including those with no obvious morphologic changes, raises the possibility that the different foci arise from distinct primary events. We hypothesize that a common event is loss of the ability to express WHV, allowing these hepatocytes to escape immune mediated cell death and to undergo clonal expansion to form distinct foci. PMID- 17078990 TI - Molecular characterization of a new porcine rotavirus P genotype found in an asymptomatic pig in Slovenia. AB - Rotaviral RNA was detected in the stool sample of an asymptomatic fattening pig at a Slovenian pig farm. To characterize the rotavirus, RT-PCR was used, employing primers specific for the VP7, VP4 and NSP4 genes. Specific products were purified and the sequencing reaction was performed for the molecular analysis of amplified genes. Nucleotide and amino acid sequences of the VP7 gene were found highly identical (85.3-88.1% and 90.7-91.6%) to G1 genotype strains. Phylogenetic and molecular analyses of the VP7 antigen regions revealed the sample to be from a new lineage of G1 genotype. In the molecular analysis of the VP4 gene, only 70.9% nucleotide (76.2% amino acid) identity was found with the most related rotavirus VP4 gene from GenBank. Following this, the NSP4 gene was also analyzed. After the phylogenetic analysis, it clustered with the NSP4 B genotype, but also seemed to represent a new lineage of this genotype. This new rotavirus strain, named P21-5, differed greatly from all rotaviruses characterized so far in all three genes analyzed. The virulence of this strain is not clear yet and has to be investigated. PMID- 17078991 TI - Acuity, crowding, reading and fixation stability. AB - People with age-related macular disease frequently experience reading difficulty that could be attributed to poor acuity, elevated crowding or unstable fixation associated with peripheral visual field dependence. We examine how the size, location, spacing and instability of retinal images affect the visibility of letters and words at different eccentricities. Fixation instability was simulated in normally sighted observers by randomly jittering single or crowded letters or words along a circular arc of fixed eccentricity. Visual performance was assessed at different levels of instability with forced choice measurements of acuity, crowding and reading speed in a rapid serial visual presentation paradigm. In the periphery: (1) acuity declined; (2) crowding increased for acuity- and eccentricity-corrected targets; and (3), the rate of reading fell with acuity-, crowding- and eccentricity-corrected targets. Acuity and crowding were unaffected by even high levels of image instability. However, reading speed decreased with image instability, even though the visibility of the component letters was unaffected. The results show that reading performance cannot be standardised across the visual field by correcting the size, spacing and eccentricity of letters or words. The results suggest that unstable fixation may contribute to reading difficulties in people with low vision and therefore that rehabilitation may benefit from fixation training. PMID- 17078992 TI - Detection of Gabor patterns of different sizes, shapes, phases and eccentricities. AB - Contrast thresholds of vertical Gabor patterns were measured as a function of their eccentricity, size, shape, and phase using a 2AFC method. The patterns were 4 c/deg and they were presented for 90 or 240 ms. Log thresholds increase linearly with eccentricity at a mean rate of 0.47 dB/wavelength. For patterns centered on the fovea, thresholds decrease as the area of the pattern increases over the entire standard deviation range of 12 wavelengths. The TvA functions are concave up on log-log coordinates. For small patterns there is an interaction between shape and size that depends on phase. Threshold contrast energy is a U shaped function of area with a minimum in the vicinity of 0.4 wavelength indicating detection by small receptive fields. Observers can discriminate among patterns of different sizes when the patterns are at threshold indicating that more than one mechanism is involved. The results are accounted for by a model in which patterns excite an array of slightly elongated receptive fields that are identical except that their sensitivity decreases exponentially with eccentricity. Excitation is raised to a power and then summed linearly across receptive fields to determine the threshold. The results are equally well described by an internal-noise-limited model. The TvA functions are insufficient to separately estimate the noise and the exponent of the power function. However, an experiment that shows that mixing sizes within the trial sequence has no effect on thresholds, suggests that the limiting noise does not increase with the number of mechanisms monitored. PMID- 17078993 TI - Comparison of the efficiency of *OH radical formation during ozonation and the advanced oxidation processes O3/H2O2 and UV/H2O2. AB - Comparison of advanced oxidation processes (AOPs) can be difficult due to physical and chemical differences in the fundamental processes used to produce OH radicals. This study compares the ability of several AOPs, including ozone, ozone+H2O2, low pressure UV (LP)+H2O2, and medium pressure UV (MP)+H2O2 in terms of energy required to produce OH radicals. Bench scale OH radical formation data was generated for each AOP using para-chlorobenzoic acid (pCBA) as an OH radical probe compound in three waters, Lake Greifensee water, Lake Zurich water, and a simulated groundwater. Ozone-based AOPs were found to be more energy efficient than the UV/H2O2 process at all H2O2 levels, and the addition of H2O2 in equimolar concentration resulted in 35% greater energy consumption over the ozone only process. Interestingly, the relatively high UV/AOP operational costs were due almost exclusively to the cost of hydrogen peroxide while the UV portion of the UV/AOP process typically accounted for less than 10 percent of the UV/AOP cost and was always less than the ozone energy cost. As the *OH radical exposure increased, the energy gap between UV/H2O2 AOP and ozone processes decreased, becoming negligible in some water quality scenarios. PMID- 17078994 TI - Characterization of benzaldehyde lyase from Pseudomonas fluorescens: A versatile enzyme for asymmetric C-C bond formation. AB - The thiamin-diphosphate-dependent enzyme benzaldehyde lyase is a very import catalyst for chemoenzymatic synthesis catalyzing the formation and cleavage of (R)-hydroxy ketones. We have studied the stability of the recombinant enzyme and some enzyme variants with respect to pH, temperature, buffer salt, cofactors and organic cosolvents. Stability of BAL in chemoenzymatic synthesis requires the addition of cofactors to the buffer. Reaction temperature should not exceed 37 degrees C. The enzyme is stable between pH 6 and 8, with pH 8 being the pH optimum of both the lyase and the ligase reaction. Potassium phosphate and Tris were identified as optimal reaction buffers and the addition of 20 vol% DMSO is useful to enhance both the solubility of aromatic substrates and products and the stability of BAL. The initial broad product range of BAL-catalyzed reactions has been enlarged to include highly substituted hydroxybutyrophenones and aliphatic acyloins. PMID- 17078995 TI - Hydroxytyrosol lipophilic analogues: enzymatic synthesis, radical scavenging activity and DNA oxidative damage protection. AB - The olive oil phenol hydroxytyrosol (3), as well its metabolite homovanillic alcohol (4), were subjected to chemoselective lipase-catalysed acylations, affording with good yield 10 derivatives (5-14) bearing C(2), C(3), C(4), C(10) and C(18) acyl chains at C-1. Hydroxytyrosol (3) and its lipophilic derivatives showed very good DPPH. radical scavenging activity. Compounds 3, 4 and their lipophilic analogues 5-14 were subjected to the atypical Comet test on whole blood cells: 3 and its analogues 5 and 6, with little hydrophobic character (logP0.05). Two-way ANOVA showed no significant differences in hardness values among groups except FS1 and FS5 which had significantly higher hardness values than FS4 and FS7 (p<0.001) and FS5 also had a higher hardness value than FS3 (p<0.05). Additionally, uncolored, FS4 and FS7 survived at 5x10(5) cycles at a stress level in the range of 60-65% of the mean biaxial flexural strength. CONCLUSION: There was no difference in flexural strength of uncolored and colored Y-TZP ceramic. The fatigue limit of uncolored, FS4 and FS7 zirconia may be defined as lying between 60 and 65% of the stress to failure. PMID- 17079009 TI - Situating alcohol use and delinquency within developmental and societal context: the case of Korean youth. AB - In this study involving self-report questionnaire data from 955 tenth-grade students in three locations within Korea, we address the meanings of alcohol use and delinquency for Korean youth. Findings (a) supported a facilitative role for alcohol, but not delinquency, with respect to perceived peer social competence; (b) indicated negative associations of both alcohol use and delinquency with parental relations, valuing academic achievement, and collectivistic values, and positive associations with friends' risk behaviours; (c) showed no relationship of these behaviours with self-esteem, coping, parental permissiveness or individualistic values; and (d) revealed that perceived benefits of alcohol use and delinquency include not only social facilitation but also exploration and assertion of independence, suggesting a potential connection, as in Western societies, between risk taking, and identity exploration. PMID- 17079010 TI - (Poly)acrylonitrile-based hydrogel as a therapeutic bulking agent in urology. AB - Vesicoureteral reflux and urinary incontinence can be treated by endoscopic injection. Injectables such as collagen and hyaluronic acid-dextranomer are commonly used, however are not offering optimal therapeutic effect yet. In the present study a novel application for poly(acrylonitrile)-based hydrogel (Hypan), already tested in other clinical indications, has been investigated for its aptitude as an injectable bulking agent. This non-degradable, extremely hydrophilic hydrogel has the capacity of a 6-fold swelling once implanted into the body, assuring the bulking effect. Deposits of this bulking agent have been implanted into the submucosal space of the bladder of 6 minipigs and the histological reaction, as well as the physical implant behavior have been studied over a time period of 6 months. The implants have developed the desired bulking effect by the above-described hydrophilic swelling. They have been enclosed into a fine fibrous tissue capsule and provoked a very limited inflammatory reaction. This study has showed that poly(acrylonitrile)-based hydrogel implants injected into the bladder are well tolerated and, due to their defined bulking ability, feasible for the treatment of vesico-ureteral reflux and urinary incontinence. To determine the therapeutic efficiency of this treatment the implants will be studied in a vesicoureteral reflux and urinary incontinence animal model. PMID- 17079011 TI - FLT3 mutation and AML/ETO in a case of Myelodysplastic syndrome in transformation corroborates the two hit model of leukemogenesis. AB - The aim of this report is to present a case of Myelodysplastic syndrome (MDS) who presented, during AML transformation, a step-wise genetic progression that corroborates the two hit model of leukemogenesis. A RCDM-RS (WHO)/RARS (FAB) patient with normal karyotype at diagnosis, evolved into AML after six months of follow up. At transformation, AML/ETO fusion was detected, although marrow blast cells were not increased until 21 days later, when FLT3-ITD was also demonstrated pointing out that the overgrowth of the FLT3/ITD clone was concomitant with the outburst of marrow blasts. These findings corroborates the two hit model of leukemogenesis in which one class of mutations (Class I) (FLT3/ITD) confers a proliferative or survival advantage to cells, and a second class of mutations (Class II) (AML/ETO) interferes with hematopoietic differentiation. PMID- 17079012 TI - Interactive cellular and physiological responses of the freshwater crab Potamonautes warreni to cadmium and microbial gill infestations. AB - Cellular and physiological responses of the freshwater crab, Potamonautes warreni, to cadmium and microbial gill infestations were investigated. Infested crabs were collected from the Mooi River, at Noordbrug, Northwest Province, South Africa in the autumn of 1995 and exposed to 0.2mg Cd(2+)L(-1) or kept unexposed under constant laboratory conditions for 7, 14 and 21 days at 24 degrees C. Cellular lesions with microbial gill infestations were persistent over 21 days and showed a similar species diversity and density as recorded in the previous summer [Schuwerack et al., 2001a. Pathological and physiological changes in the South African freshwater crab, P. warreni Calman induced by microbial gill infestations. J. Invertebrate Pathol. 77(4), 259-269]. With Cd exposure the microbial community disappeared and crabs showed a higher mean but wider range in ammonia excretion and lower mean O:N ratio after 7 days. After 14 days exposure to Cd, time- and stressor-specific cellular and physiological responses included degeneration of oxygen and gas exchange gill tissues and a mean reduced growth rate, ion exchange, a significantly reduced O:N ratio (PMg>Zn, and the average concentrations of lowest metallic elements was Pb>Cu>Mn>Cr>Cd. The results also indicated that the concentration of metallic element Cu increased as the averaged concentrations of metallic element Zn and Mn increased during the past 10 years in Asian countries. PMID- 17079078 TI - Uptake and translocation of Cd in different rice cultivars and the relation with Cd accumulation in rice grain. AB - The variations among six rice cultivars in cadmium (Cd) uptake and translocation were investigated with pot soil experiments. The results showed that only a very small portion (0.73%) of Cd absorbed by rice plant was transferred into grain. With regard to plant total Cd uptake, Cd concentrations and quantity accumulations in roots, stems and leaves, the differences among the cultivars (between the largest one and the smallest one) were less than one time. But for Cd concentrations and Cd quantity accumulations in the grains, the differences were more than five and eight times, respectively. With respect to Cd distribution portions in plant organs, the diversities among the cultivars were also small in roots, stems and leaves, but much larger in grains. Grain Cd concentrations correlated positively and significantly (P<0.01) with Cd quantity accumulations in plant, Cd distribution ratios to aboveground parts, and especially with Cd distribution ratios from aboveground parts to the grain. The results indicated that Cd concentration in rice grain was governed somewhat by plant Cd uptake and the transport of Cd from root to shoot, and in a greater extent, by the transport of Cd from shoot to grain. Cd was not distributed evenly in different products after rice grain processing. The average Cd concentration in cortex (embryo) was five times more than that in chaff and polished rice. With regard to Cd quantity accumulation in the products, near 40% in cortex (embryo), 45% in polished rice and 15% in chaff averagely. PMID- 17079079 TI - Synthesis and sulfate ion-exchange properties of a hydrotalcite-like compound intercalated by chloride ions. AB - To reduce the generation of hydrogen sulfide gas from sulfate ions, we synthesized a layered double hydroxide hydrotalcite-like compound intercalated with chloride ions in the interlayer (HT-Cl) using a coprecipitation reaction. The resultant HT-Cl material had a Mg/Al molar ratio of approximately 2, and the molar fraction of the chloride ions on the intercalated anion layer of the HT was 0.90. A higher molar fraction of chloride ions can be inserted into the interlayers of the HT by increasing the initial concentration ratio of chloride ions to aluminum ions [Cl(-)](0)/[Al(3+)](0) in the solution. Approximately 98% of the sulfate ions in the solution were removed within 10 min after adding 1.12 times the stoichiometric quantity of synthesized HT-Cl, which contained a Mg/Al molar ratio of 2 equal to a solution containing a 12 mM calcium sulfate solution. PMID- 17079080 TI - Family-based and case-control study of DRD2, DAT, 5HTT, COMT genes polymorphisms in alcohol dependence. AB - The paper focuses on such candidate gene polymorphisms that alter alcoholism related intermediate phenotypes including: dopaminergic system polymorphic variants (DRD2 -141C Ins/Del in promoter region, exon 8 and DRD2 TaqI A and DAT 40bp VNTR genes polymorphisms) that cause predisposition to severe alcoholism (haplotype Ins/G/A2); COMT Val158Met gene polymorphism related to differences in executive cognitive function and 5-HTT gene promoter polymorphism, which alters stress response and affects anxiety and dysphoria. The transmission disequilibrium test (TDT) was used in the study. One hundred Polish families with alcohol dependence were recruited. The control subjects for the case-control study were 196 ethnically and gender matched healthy individuals. It was found that DRD2 TaqIA and DAT gene polymorphisms contained statistically significant differences in allele transmission. In the homogenous subgroups of patients with early onset and with withdrawal complications a statistically significant preferential A2 allele transmission was found in DRD2 TaqIA gene polymorphism. The alleles and genotypes distribution of the investigated polymorphisms did not differ significantly between the alcoholics and the controls in the case-control study. The results confirmed the fact that the candidate genes (DRD2 and DAT) are partially responsible for the development of alcohol dependence. The results are also in agreement with the hypothesis that there are various subtypes of alcohol dependence, which differ depending on their genetic background. Meanwhile, the currently available pharmacological therapies for alcoholism treatment are effective in some alcoholics but not for all of them. Some progress has been made in elucidating pharmacogenomic responses to drugs, particularly in the context of Clonninger and Lesch typology classification system for alcoholics. PMID- 17079081 TI - A method of combining STEM image with parallel beam diffraction and electron optical conditions for diffractive imaging. AB - We describe a method of combining STEM imaging functionalities with nanoarea parallel beam electron diffraction on a modern TEM. This facilitates the search for individual particles whose diffraction patterns are needed for diffractive imaging or structural studies of nanoparticles. This also lays out a base for 3D diffraction data collection. PMID- 17079082 TI - Design of a microfabricated, two-electrode phase-contrast element suitable for electron microscopy. AB - A miniature electrostatic element has been designed to selectively apply a 90 degrees phase shift to the unscattered beam in the back focal plane of the objective lens, in order to realize Zernike-type, in-focus phase contrast in an electron microscope. The design involves a cylindrically shaped, biased-voltage electrode, which is surrounded by a concentric grounded electrode. Electrostatic calculations have been used to determine that the fringing fields in the region of the scattered electron beams will cause a negligible phase shift as long as the ratio of electrode length to the transverse feature size is greater than 5:1. Unlike the planar, three-electrode einzel lens originally proposed by Boersch for the same purpose, this new design does not require insulating layers to separate the biased and grounded electrodes, and it can thus be produced by a very simple microfabrication process. Scanning electron microscope images confirm that mechanically robust devices with feature sizes of approximately 1 microm can be easily fabricated. Preliminary experimental images demonstrate that these devices do apply a 90 degrees phase shift between the scattered and unscattered electrons, as expected. PMID- 17079083 TI - Subcutaneous compartment syndrome. PMID- 17079084 TI - Who burned and how to prevent? Identification of risk for and prevention of burns among epileptic patients. PMID- 17079085 TI - Long-term risk factors for impaired burn-specific health and unemployment in patients with thermal injury. AB - The success of acute burn therapy has led to an increased demand for high-quality rehabilitation. When optimizing burn care programs, knowledge of long-term risk factors associated with impaired health and unemployment of the patient may be significant. The health and work status of 95 patients (82.1% males; mean age 43.7 (S.D.: 14.5) years; mean total body surface burn 18.5 (S.D.: 14.2) % were assessed 47.0 (S.D.: 23.8) months after injury, using the Norwegian version of the abbreviated burn-specific health scale (BSHS-N) and a questionnaire asking for socio-demographic and medical characteristics. A regression model demonstrated that the BSHS-N total score was significantly reduced by chronic pain (P<0.001), psychological illness (P<0.001), and living alone (P=0.030), as well as full-thickness facial (P=0.011) and foot (P=0.013) burns. Unemployment was significantly associated with housing and economic problems (P=0.001), chronic pain (P=0.001), the extent of full-thickness injury (P=0.005), the presence of deformities (P=0.037), the number of operations (P=0.001) and the length of hospital stay (P=0.016). Thus, socio-demographic factors, non-burn related morbidity and the injury itself significantly impaired long-term physical and psychosocial health and work status. PMID- 17079086 TI - Delayed reepithelialization and scarring deregulation following drug-induced toxic epidermal necrolysis. AB - A 51-year-old Caucasian woman developed severe drug-induced toxic epidermal necrolysis (TEN) due to allopurinol. The withdrawal of the culprit drug was unfortunately delayed, and dramatic retardation of reepithelialization was observed. At that stage of disease evolution, an inflammatory cell infiltrate was present in the dermis. Coverage of eroded lesions by frozen cultured keratinocyte allografts failed to hasten reepithelialization compared to ungrafted sites. This unusual protracted TEN evolution was followed by the development of extensive hypertrophic and keloid scars. Several biopsies were taken over 6 months. The histologic presentation of the grafted and ungrafted eroded scar tissues looked similar. Both the number and size of the Factor XIIIa-positive dermal dendrocytes, as well as the number of alpha-actin-positive myofibroblasts showed a marked increase between weeks 2 and 12 after grafting. They were reduced after 6 months when the scarring process was stabilized. alpha1 [IV] collagen was never expressed over the eroded scars. Similar to burn patients, delayed reepithelialization might be a risk factor for abnormal scarring in TEN. Cultured keratinocyte allograft apparently offered no improvement in reepithelialization and did not prevent abnormal scarring in this TEN patient. PMID- 17079087 TI - Apoptotic cells in cutaneous adnexa of burned patients. PMID- 17079088 TI - Post-operative analgesia for open wounds and painful dressings. PMID- 17079089 TI - The effect of treating infected skin grafts with Acticoat on immune cells. AB - A study was conducted to determine the effect of Acticoat placed on an infected skin graft on parameters of immunity. Two partial thickness wounds (2 cm x 4 cm) were created on the dorsal midline of Hartley guinea pigs (n=28). Wounds were covered with autologous skin graft and maintained either aseptically (Noninoculated, n=8), inoculated with Staphylococcus aureus (Surgery-Inoculated, n=8) with or without Acticoat bandage (Surgery-Inoculated-Acticoat, n=6). Five days later, splenocytes and blood were collected to estimate natural killer cell (NK) cytotoxicity, proliferative response to T and B cell mitogens and neutrophil oxidative burst. Animals that did not undergo surgery were included as a nonsurgery control group. [(3)H]-thymidine incorporation in response to a variety of T and B cell mitogens was significantly lower for all groups undergoing surgery compared to the nonsurgery control group (p<0.0001) and no additional effect was observed on this immune measure by applying the Acticoat bandage. The Surgery-Inoculated-Acticoat group exhibited greater NK cytotoxic activity (as assessed as the ability to lyse K562 tumor cells) compared to the Surgery Inoculated group (p<0.006). The Surgery-Inoculated-Acticoat group had higher neutrophil oxidative burst at 5 min post stimulation, but was not different from controls after 15 min. In conclusion, the application of an Acticoat bandage to an inoculated surgery wound did not alter the low cell-mediated immune response that followed surgery, but appeared to increase parameters (NK cytotoxic activity and neutrophil function) of innate immunity. PMID- 17079090 TI - Effects of pictures and textual arguments in sun protection public service announcements. AB - BACKGROUND: The effect of public service announcements aimed at promoting primary prevention of skin cancer may be limited by superficial cognitive processing. The use of both pictures and textual arguments in sun protection public service announcements were evaluated for their potentially beneficial effects on judgment, cognitive processing and persuasiveness. METHODS: In a 2 x 2 factorial experimental design individuals were shown public service announcements that advocated the advantages of sun protection measures in different versions in which a picture was present or not present and a textual argument was present or not present. The 159 participants were randomly assigned to one of four conditions. In each condition, participants were shown 12 different public service announcements designed according to the condition. Participants judged each public service announcement on attractiveness, credibility, clarity of communication and the required amount of reflection. After the judgment task, they completed a questionnaire to assess knowledge, perceived advantages and disadvantages of sun protection and intended use of sun protection measures. RESULTS: Pictures enhanced attractiveness, but diminished comprehension. Textual arguments enhanced attractiveness, credibility and comprehension. Pictures as well as textual arguments increased knowledge of sun protection measures. CONCLUSION: Pictures and textual arguments in public service announcements positively influence the individual's perception of the advantages of sun protection methods and the advantages of their adoption. PMID- 17079091 TI - Precancer: a conceptual working definition -- results of a Consensus Conference. AB - BACKGROUND: Precancers are lesions that precede the appearance of invasive cancers. The successful prevention or treatment of precancers has the potential to eliminate deaths due to cancer. METHODS: A National Cancer Institute-sponsored Conference on Precancer was convened on November 8-9, 2004, at The George Washington University Medical Center, Washington, DC. A definition of precancers was developed over 2 days of Conference discussions. RESULTS: The following five criteria define a precancer: (1) evidence must exist that the precancer is associated with an increased risk of cancer; (2) when a precancer progresses to cancer, the resulting cancer arises from cells within the precancer; (3) a precancer differs from the normal tissue from which it arises; (4) a precancer differs from the cancer into which it develops, although it has some, but not all, of the molecular and phenotypic properties that characterize the cancer; (5) there is a method by which the precancer can be diagnosed. CONCLUSIONS: The Conference participants developed a general definition for precancers that would provide a consistent and clinically useful way of distinguishing precancers from all other types of lesions. It was recognized that many precancerous lesions may not meet this strict definition, but the group felt it was necessary to define criteria that will help standardize clinical and biological studies. Furthermore, a set of defining criteria for putative precancer lesions will permit pathologists to build a diagnostically useful taxonomy of precancers based on specified clinical and biological properties. Precancers thus characterized can be classified into clinically relevant sub-groups based on shared properties (i.e. biomarkers, oncogenes, common metabolic pathways, responses to therapy, etc.). Publications that introduce newly described precancer entities should describe how each of the five defining criteria apply. This manuscript reviews the proposed definition of precancers and suggests how pathologists, oncologists and cancer researchers may determine when these criteria are satisfied. PMID- 17079092 TI - Training delays reduce the choose-short effect with keylight, but not with food, duration samples in pigeons. AB - Pigeons were trained to match 2- and 8-s food samples. The delay on training trials was either 0s (group 0sF), 5s (group 5sF), or varied between 2 and 8s (M=5s, group 5sV). Testing at a delay that exceeded the training delay by 15s in each group revealed a robust choose-short effect in each group. The same pigeons then reacquired a previously trained matching task involving 2- and 8-s keylight samples. Different comparison stimuli were used on food-sample and keylight sample trials. The delay on training trials was the same on both food- and keylight-sample trials. Extended-delay testing revealed a robust choose-short effect in all three groups when the durations were conveyed by food presentations, but only group 0sF revealed a choose-short effect when the durations were conveyed by keylight presentations. Hence, training with a nonzero delay, whether fixed or variable, reduces the choose-short effect with keylight durations but not with food durations. It was concluded that at least some of the psychological processes mediating performance differ as a function of the event that conveys the duration. PMID- 17079093 TI - Severe suppression of Frzb/sFRP3 transcription in osteogenic sarcoma. AB - Deciphering the molecular basis of cancer is critical for developing novel diagnostic and therapeutic strategies. To better understand the early molecular events involving osteogenic sarcoma (OGS), we have initiated a program to identify potential tumor suppressor genes. Expression profiling of total RNA from ten normal bone cell lines and eleven OGS-derived cell lines by microarray showed 135-fold lower expression of FRZB/sFRP3 mRNA in OGS cells compared to bone cells; this down-regulation of Frzb/sFRP3 mRNA expression was found to be serum independent. Subsequently, fourteen OGS biopsy specimens showed nine-fold down regulation of Frzb/sFRP3 mRNA expression compared to expression in eight normal bone specimens as determined by microarray. FRZB /sFRP3 protein level was also found to be at a very low level in 4/4 OGS cell lines examined. Quantitation by RT-PCR indicated approximately 70% and approximately 90% loss of Frzb/sFRP3 mRNA expression in OGS biopsy specimens and OGS-derived cell lines respectively, compared to expression in bone (p<0.0001). Hybridization experiments of a cDNA microarray containing paired normal and tumor specimens from nineteen different organs did not show any significant difference in the level of Frzb/sFRP3 mRNA expression between the normal and the corresponding tumor tissues. Exogenous expression of FRZB/sFRP3 mRNA in two OGS-derived cell lines lacking endogenous expression of the mRNA produced abundant mRNA from the exogenous gene, eliminating degradation as a possibility for very low level of FRZB/sFRP3 mRNA in OGS specimens. Results from PCR-based experiments suggest that the FRZB/sFRP3 gene is not deleted in OGS cell lines, however, karyotyping shows gross abnormalities involving chromosome 2 (location of the FRZB gene) in five of twelve OGS-derived cell lines. Together, these data suggest a tumor-suppressive potential for FRZB/sFRP3 in OGS. PMID- 17079095 TI - Selective inhibition of Alu retrotransposition by APOBEC3G. AB - The non-LTR retrotransposon LINE-1 (L1) comprises approximately 17% of the human genome, and the L1-encoded proteins can function in trans to mediate the retrotransposition of non-autonomous retrotransposons (i.e., Alu and probably SVA elements) and cellular mRNAs to generate processed pseudogenes. Here, we have examined the effect of APOBEC3G and APOBEC3F, cytidine deaminases that inhibit Vif-deficient HIV-1 replication, on Alu retrotransposition and other L1-mediated retrotransposition processes. We demonstrate that APOBEC3G selectively inhibits Alu retrotransposition in an ORF1p-independent manner. An active cytidine deaminase site is not required for the inhibition of Alu retrotransposition and the resultant integration events lack G to A or C to T hypermutation. These data demonstrate a differential restriction of L1 and Alu retrotransposition by APOBEC3G, and suggest that the Alu ribonucleoprotein complex may be targeted by APOBEC3G. PMID- 17079096 TI - A virus vector based on Canine Herpesvirus for vaccine applications in canids. AB - Canine Herpesvirus (CHV) is being developed as a virus vector for the vaccination of European red foxes. However, initial studies using recombinant CHV vaccines in foxes revealed viral attenuation and lack of antibody response to inserted foreign antigens. These findings were attributed both to inactivation of the thymidine kinase (TK) gene and excess foreign genetic material in the recombinant viral genome. In this study, we report an improved CHV-bacterial artificial chromosome (BAC) vector system designed to overcome attenuation in foxes. A non essential region was identified in the CHV genome as an alternative insertion site for foreign genes. Replacement of a guanine/cytosine (GC)-rich intergenic region between UL21 and UL22 of CHV with a marker gene did not change growth behaviour in vitro, showing that this region is not essential for virus growth in cell culture. We subsequently produced a CHV-BAC vector with an intact TK gene in which the bacterial genes and the antigen expression cassette were inserted into this GC-rich locus. Unlike earlier constructs, the new CHV-BAC allowed self excision of the bacterial genes via homologous recombination after transfection of BACs into cell culture. The BAC-CHV system was used to produce a recombinant virus that constitutively expressed porcine zona pellucida subunit C protein between the UL21 and UL22 genes of CHV. Complete self-excision of the bacterial genes from CHV was achieved within one round of replication whilst retaining antigen gene expression. PMID- 17079097 TI - Assessment and support of the preterm circulation. AB - There are no clinical outcome data on which to base recommendations on how to assess and support the preterm circulation. Current standards are derived from an assumed proportionality between systemic and organ blood flow and mean blood pressure. Our study of central measures of systemic blood flow suggests preterm haemodynamics are more complex than this. Low systemic blood flow is common in the first 24 h after birth in very preterm babies and is not necessarily reflected by low blood pressure. The causes of this low systemic blood flow are complex but may relate to maladaptation to high extrauterine systemic (and sometimes pulmonary) vascular resistance. After day 1, hypotensive babies are more likely to have normal or high SBF reflecting vasodilatation. Empirically, inotropes that reduce afterload (such as dobutamine) may be more appropriate in the transitional period, while those with more vasoconstrictor actions (such as dopamine) may be more appropriate later on. Defining the haemodynamic in an individual baby needs both blood pressure and echocardiographic measures of systemic blood flow. Research in this area needs to move beyond just demonstrating changes in physiological variables to showing improvements in important clinical outcomes. PMID- 17079098 TI - The impact of maternal smoking on fetal and infant growth. AB - BACKGROUND: Low birth weight is associated with accelerated postnatal growth and adverse adult health outcomes. Maternal smoking is a major risk factor for low birth weight. This study aims to assess: Pre- and postnatal growth associated with maternal smoking compared to other risk factors for low birth weight. The effect of reduction of maternal smoking on growth. SUBJECTS: A cohort (n=269) followed with ultrasound measurements in the third trimester and postnatal anthropometric measurements until 6 months of age. Mothers were interviewed about their smoking habits at 18 and 28 weeks of pregnancy. RESULTS: Maternal smoking was associated with a greater reduction in birth length SDS than other causes of equally reduced birth weight (mean difference: -0.25 SDS, P=0.013). The adjustment of gestational age, based on bi-parietal diameter at an early dating scan, indicated that mothers who reduced smoking carried smaller fetuses than mothers who continued to smoke heavily (mean difference=2.6 days, P=0.012). Birth weights in these two groups were similar (P=0.87). However at 3 months of age, reduced smoking was associated with lower weight (mean difference=-0.38 SDS, P=0.045). CONCLUSIONS: Maternal smoking was associated with a reduction of linear growth, which was more marked than that of other risk factors, and which seemed to occur before the 3rd trimester. The results indicated a beneficial effect of reduction of smoking upon third trimester growth, and that the decision to reduce smoking in mid-pregnancy may be influenced by early fetal size. PMID- 17079099 TI - Transient hypothyroxinaemia in preterm infants. AB - Thyroid hormones are required for normal development of the brain. Transient hypothyroxinaemia is the most common thyroid dysfunction in preterm infants and is defined by temporary low levels of T4, T3 and normal or low TSH. Low T4 levels in preterm infants are associated with persistent neurodevelopmental deficits in cognitive and motor function. Thyroid hormone substitution trials to date are underpowered and show inconsistent results; the question remains -- are low T4 levels simply an epiphenomenon? The aetiology of transient hypothyroxinaemia is multifactorial and the components amenable to correction form the basis of the therapeutic strategy: rectification of iodine deficiency in parenteral nutrition; a reduction of non-thyroidal illnesses and attenuation of their severity; and substitution of drugs that interfere with the hypothalamic-pituitary-thyroid axis. Thyroxine substitution therapy should only be done in the context of clinical trials and only in those infants who are hypothyroxinaemic. PMID- 17079100 TI - Evaluation of HO-1-u-1 cell line as an in vitro model for sublingual drug delivery involving passive diffusion--Initial validation studies. AB - The aim of this study was to provide preliminary validation of a new sublingual mucosal cell line (HO-1-u-1) for use as in vitro sublingual drug delivery screening of compounds involving passive diffusion. HO-1-u-1 cells were seeded on cell culture inserts. The ultrastructure and integrity of cell layers, inter passage variation and directionality of drug transport, and apparent permeability coefficient (P(app)) of eight beta-blockers (representing compounds involving passive diffusion) were determined. HO-1-u-1 cells grown on inserts formed stratified and epithelial-like structure and maintained the typical histological features of normal human sublingual epithelium. The maximal integrity of the cell layer was reached in 23 days. No significant inter-passage variation was found at the passages ranging from 2 to 11 when measured by radiolabeled transcellular and paracellular markers (testosterone and mannitol, respectively). Bidirectional transport studies confirmed the passive diffusion as the mechanism of transport for these markers. The P(app) of eight beta-blockers across HO-1-u-1 cell culture ranged from 2.89+/-0.17 to 6.37+/-0.37x10(-6)cm/s and correlated well to the P(app) obtained from porcine sublingual mucosa (r(2)=0.647 and 0.83 when excluding propranolol). The above results indicate that the HO-1-u-1 cells grown on inserts may offer as a potentially in vitro model for screening sublingual drug permeation involving passive diffusion. PMID- 17079102 TI - High-frequency tinnitus without hearing loss does not mean absence of deafferentation. AB - A broad consensus within the neuroscience of tinnitus holds that this audiologic condition is triggered by central deafferentation, mostly due to cochlear damage. The absence of audiometrically detectable hearing loss however poses a challenge to this rather generalizing assumption. The aim of this study was therefore to scrutinize cochlear functioning in a sample of tinnitus subjects audiometrically matched to a normal hearing control group. Two tests were applied: the Threshold Equalizing Noise (TEN) test and a pitch scaling task. To perform well on both tasks relatively normal functioning of inner hair cells is a requirement. In the TEN test the tinnitus group revealed a circumscribed increment of thresholds partially overlapping with the tinnitus spectrum. Abnormal slopes were observed in the pitch scaling task which indicated that tinnitus subjects, when presented with a high-frequency stimulus, relied heavily on input derived from lower frequency inner hair cells (off-frequency listening). In total both results argue for the presence of a deafferentation also in tinnitus subjects with audiometrically normal thresholds and therefore favour the deafferentation assumption posed by most neuroscientific theories. PMID- 17079101 TI - Preparation of large porous biodegradable microspheres by using a simple double emulsion method for capreomycin sulfate pulmonary delivery. AB - The aim of this work was to evaluate if a simple double-emulsion method could be used for developing a new formulation of large porous microspheres (MS) potentially useful for capreomycin sulfate (CS) pulmonary delivery. Poly(DL lactide-co-glycolide) was used for MS preparation. A simple W/O/W double emulsion/solvent evaporation preparation method was employed and MS were characterized by UV spectrophotometry, particle size, and scanning electron microscopy. A computer-generated response surface method (RSM) was employed to evaluate % drug content, volume mean diameter (VMD), and span upon variation of two numeric and two categorical factors. MS size distribution was found to be strongly affected by the homogenization method and the type of emulsifier employed. Mean diameters ranged from 1 to 20 microm. The MS presented a proper morphology, with a highly porous interior and a rough surface. Peptide content ranged between 1 and 20%. The region of optimality was referred to as a low VMD and span values, and a high drug content. The best results were found when using a 20% loading, 19.8-3.2 dichloromethane/acetone ratio, ultraturrax mixing, and HPMC as emulsifier. The double-emulsion method allowed the preparation of CS loaded large porous MS having suitable characteristics to match respirability requirements. The use of RSM helped to establish the conditions to obtain formulations potentially useful for a possible CS pulmonary delivery, by using a simple preparation method with a consistent time, cost, and material saving. PMID- 17079103 TI - Voltage-gated and two-pore-domain potassium channels in murine spiral ganglion neurons. AB - The systematically varied firing features of spiral ganglion neurons provide an excellent model system for the exploration of how graded ion channel distributions can be used to organize neuronal firing across a population of neurons. Elucidating the underlying mechanisms that determine neuronal response properties requires a complete understanding of the combination of ion channels, auxiliary proteins, modulators, and second messengers that form this highly organized system in the auditory periphery. Toward this goal, we built upon previous studies of voltage-gated K+-selective ion channels (Kv), and expanded our analysis to K+-selective leak channels (KCNK), which can play a major role in setting the basic firing characteristics of spiral ganglion neurons. To begin a more comprehensive analysis of Kv and KCNK channels, a screening approach was employed. RT-PCR was utilized to examine gene expression, the major results of which were confirmed with immunocytochemistry. Initial studies validated this approach by accurately detecting voltage-dependent K+ channels that were documented previously in the spiral ganglion. Furthermore, an additional channel type within the Kv3 family, Kv3.3, was identified and further characterized. The major focus of the study, however, was to systematically examine gene expression levels of the KCNK family of K+-selective leak channels. These channel types determine the resting membrane potential which has a major impact on setting the level of neuronal excitation. TWIK-1, TASK-3, TASK-1, and TREK-1 were expressed in the spiral ganglion; TWIK-1 was specifically localized with immunocytochemistry to the neuronal somata and initial processes of spiral ganglion neurons in vitro. PMID- 17079104 TI - Physiology, pharmacology and plasticity at the inner hair cell synaptic complex. AB - This report summarizes recent neuropharmacological data at the IHC afferent/efferent synaptic complex: the type of Glu receptors and transporter involved and the modulation of this fast synaptic transmission by the lateral efferents. Neuropharmacological data were obtained by coupling the recording of cochlear potentials and single unit of the auditory nerve with intra-cochlear applications of drugs (multi-barrel pipette). We also describe the IHC afferent/efferent functioning in pathological conditions. After acoustic trauma or ischemia, acute disruption of IHC-auditory dendrite synapses are seen. However, a re-growth of the nerve fibres and a re-afferentation of the IHC were completely done 5 days after injury. During this synaptic repair, multiple presynaptic bodies were commonly found, either linked to the membrane or "floating" in ectopic positions. In the meantime, the lateral efferents directly contact the IHCs. The demonstration that NMDA receptors blockade delayed the re growth of neurites suggests a neurotrophic role of NMDA receptors in pathological conditions. PMID- 17079105 TI - Effect of Catha edulis foresk (khat) extracts on male rat sexual behavior. AB - Khat chewing is a widespread habit that has a deep-rooted socio-cultural tradition in East Africa and in the Middle East. Although a number of investigations have been carried out using cathinone, the psychoactive component of khat, these may not wholly reflect the behavioral effects observed after administering khat in a dosage similar to those used traditionally. The aim of the present study was to evaluate the effect of sub-chronically administered khat extract with or without alcohol on sexual behavior in male rats. Adult albino wistar male rats were administered either with khat extracts (100, 200, 400mg/kg), amphetamine (1mg/kg), sildenafil (1mg/kg), ethanol (2ml/kg of 2% and 10%), or a combination of khat and ethanol (2%+10%) by intragastric gavage orally for 15 days. Khat (400mg/kg) treated rats demonstrated a statistically significant increase in all sexual parameters except in mounting frequency, intercopulatory interval and copulatory efficiency. Whereas, khat (200mg/kg) treated rats showed a statistically significant increase only in ejaculation latency (P<0.01). In marked contrast, low dose (100mg/kg) of khat extract was found to significantly reduce both mount latency (P<0.05) and intromission latency P<0.01) thereby enhancing sexual motivation/arousal in male rats. Similar results were obtained when khat extract (200mg/kg) and ethanol (10%) were administered concomitantly despite the inhibitory effect observed in male sexual behavior when administered alone. From the present study it can be concluded that higher doses of the extract inhibit sexual behavior in male rats. In contrast, low dose of the extract as well as the concurrent administration of the extract followed by ethanol was found to enhance male rat sexual motivation/arousal. PMID- 17079106 TI - Multi-detector CT imaging in the postoperative orthopedic patient with metal hardware. AB - Multi-detector CT imaging (MDCT) becomes routine imaging modality in the assessment of the postoperative orthopedic patients with metallic instrumentation that degrades image quality at MR imaging. This article reviews the physical basis and CT appearance of such metal-related artifacts. It also addresses the clinical value of MDCT in postoperative orthopedic patients with emphasis on fracture healing, spinal fusion or arthrodesis, and joint replacement. MDCT imaging shows limitations in the assessment of the bone marrow cavity and of the soft tissues for which MR imaging remains the imaging modality of choice despite metal-related anatomic distortions and signal alteration. PMID- 17079107 TI - Nature of the main contaminant in the drug primaquine diphosphate: SFC and SFC-MS methods of analysis. AB - The drug primaquine diphosphate is used for causative treatment of malaria. Using HPLC-MS and GC-MS, this research group was previously able to show that the main contaminant of primaquine is the positional isomer quinocide [I. Brondz, D. Mantzilas, U. Klein, D. Ekeberg, E. Hvattum, M.N. Lebedeva, F.S. Mikhailitsyn, G.D. Soulimanov, J. Roe, J. Chromatogr. B: Anal. Technol. Biomed. Life Sci. 800 (2004) 211-223; I. Brondz, U. Klein, D. Ekeberg, D. Mantzilas, E. Hvattum, H. Schultz, F. S. Mikhailitsyn, Asian J. Chem. 17 (2005) 1678-1688]. Primaquine and quinocide are highly toxic substances which can have a number of side effects upon use in medical treatment. A standard for quinocide is not typically commercially available. In the present work, supercritical fluid chromatography mass spectrometry (SFC-MS) with two different columns was used to achieve a shorter analysis time for the separation between the positional isomers quinocide and primaquine in primaquine diphosphate and to elucidate additional information about differences in their MS fragmentation. Unlike using HPLC-MS, it was possible to achieve the differential fragmentation of positional isomers at branching points using the SFC-MS technique. The desired short analysis time was achieved using SFC equipped with a Discovery HS F5 column and the differential fragmentation of positional isomers during SFC-MS provides information on the differences in the structure of these substances. Using a Chiralpak AD-H chiral column, it was possible to resolve the enantiomers in primaquine and separate quinocide from those enantiomers. PMID- 17079108 TI - Simultaneous analysis of eight bioactive compounds in Danning tablet by HPLC-ESI MS and HPLC-UV. AB - A high performance liquid chromatography (HPLC) coupled with electrospray tandem mass spectrometry (ESI-MS) and ultraviolet detector (UV) has been developed for the simultaneous analysis of eight bioactive compounds in Danning tablet (including hyperin, hesperidin, resveratrol, nobiletin, curcumine, emodin, chrysophanol, and physcion), a widely used prescription of traditional Chinese medicine (TCM). The chromatographic separation was performed on a ZORBAX Extend C(18) analytical column by gradient elution with acetonitrile and formate buffer (containing 0.05% formic acid, adjusted with triethylamine to pH 5.0) at a flow rate of 0.8 ml/min. The eight compounds in Danning tablet were identified and their MS(n) fractions were elucidated by using HPLC-ESI-MS, and the contents of these compounds were determined by using HPLC-UV method. The standard calibration curves were linear between 5.0 and 100 microg/ml for hyperin, 10-200 microg/ml for hesperidin, 1.0-150 microg/ml for resveratrol, 2.0-120 microg/ml for nobiletin, 2.0-225 microg/ml for curcumine, 20-300 microg/ml for emodin, 2.0-200 microg/ml for chrysophanol, and 20-250 microg/ml for physcion with regression coefficient r(2)>0.9995. The intra-day and inter-day precisions of this method were evaluated with the R.S.D. values less than 0.7% and 1.3%, respectively. The recoveries of the eight investigated compounds were ranged from 99.3% to 100.2% with R.S.D. values less than 1.5%. This method was successfully used to determine the 8 target compounds in 10 batches of Danning tablet. PMID- 17079109 TI - Experimental design optimization for the ICP-AES determination of Li, Na, K, Al, Fe, Mn and Zn in human serum. AB - A chemometric approach based on experimental design and desirability functions was used to develop and validated a method for the determination of some metals of biological concern by a fast sequential ICP-AES. The elements considered are Al, Fe, Mn, Zn, Li, Na and K. The experimental design was used to investigate the effects of three instrumental most crucial parameters, such as sheath gas flow rate, pump speed and auxiliary gas flow rate. In order to improve the multielemental analysis speed, although a sequential instrument allows the use of a separate parameter set for each wavelength, regression models and desirability functions were applied to find the experimental conditions providing the highest global sensitivity. Validation was performed in terms of limits of detection (LOD), limits of quantitation (LOQ), linearity, precision and recovery. By using the 167.02 nm wavelength, aluminium LOD was 0.5 microg L(-1) while the highest LOD was found for K (65 microg L(-1)). A linear range of at least three orders of magnitude was statistically demonstrated for each element. Precision was evaluated by testing two concentration levels, and good results in terms of intra day repeatability were obtained, with R.S.D. values lower than 4.1% at the lowest concentration level. Lacking a suitable certified reference material, trueness was estimated using the recovery rate on fortified samples. The validated method was then used in the quantification of the elements considered in a serum sample. PMID- 17079110 TI - Inactivation of lactate dehydrogenase by several chemicals: implications for in vitro toxicology studies. AB - Lactate dehydrogenase (LDH) release is frequently used as an end-point for cytotoxicity studies. We have been unable to measure LDH release during studies using para-aminophenol (PAP) in LLC-PK(1) cells. When LLC-PK(1) cells were incubated with either PAP (0-10 mM) or menadione (0-1000 microM), viability was markedly reduced when assessed by alamar Blue or total LDH activity but not by release of LDH into the incubation medium. In addition, we incubated cells with PAP or menadione and compared LDH activity using two different assays. Both assays confirmed our observation of decreased LDH activity in cell lysates without corresponding increases in LDH activity in incubation media. Using purified LDH and 10 mM PAP, we found that PAP produced loss of LDH activity that was inversely proportional to the amount of LDH initially added. In additional experiments, we incubated 0.5 units of LDH for 1 h with varying concentrations of PAP, menadione, hydrogen peroxide (H(2)O(2)) or cisplatin. All four chemicals produced concentration-dependent decreases in LDH activity. In previous experiments, inclusion of antioxidants such as reduced glutathione (GSH) and ascorbate protected cells from PAP toxicity. GSH (1 mM) preserved LDH activity in the presence of toxicants while ascorbate (1 mM) only prevented LDH loss induced by PAP. These studies suggest that LDH that is released into the incubation medium is susceptible to degradation when reactive chemicals are present. PMID- 17079111 TI - Performance of Spanish speakers on the Mattis dementia rating scale (MDRS). AB - This paper examined whether education-, age-, and gender-matched Spanish- and English-speaking normals (n=30 pairs) had comparable scores on the Mattis dementia rating scale (MDRS). It provides preliminary normative data on Spanish speaking volunteers aged 55-89 years old (n=54). It also compared the MDRS total score with its memory subscale score and the mini-mental state examination (MMSE) score on sensitivity and specificity for distinguishing normals from patients with dementia (n=61). Spanish-speaking normals scored significantly lower than English-speaking normals on MDRS total and its attention, conceptualization, and memory subscales. The area under the receiver operating characteristic curve distinguishing normals from patients with dementia was not significantly different among the MDRS total, its memory subscale, and the MMSE. We conclude that (a) the norms based on English-speaking individuals are not appropriate for use with Spanish-speaking individuals, and (b) to screen for dementia, the shorter MDRS Memory subscale and the MMSE are as good as the entire MDRS. PMID- 17079112 TI - Psychometric characteristics of the Mobility Inventory in a longitudinal study of anxiety disorders: replicating and exploring a three component solution. AB - Psychometric characteristics of the Mobility Inventory (MI) were examined in 216 outpatients diagnosed with panic disorder with agoraphobia participating in a longitudinal study of anxiety disorders. An exploratory principal components analysis replicated a three-component solution for the MI reported in prior studies, with components corresponding to avoidance of public spaces, avoidance of enclosed spaces, and avoidance of open spaces. Correlational analyses suggested that the components tap unique but related areas of avoidance that were remarkably stable across periods of 1, 3, and 5 years between administrations. Implications of these results for future studies of agoraphobia are discussed. PMID- 17079113 TI - Clonidine reduces hypersensitivity and alters the balance of pro- and anti inflammatory leukocytes after local injection at the site of inflammatory neuritis. AB - Perineural alpha2-adrenoceptor activation relieves hypersensitivity induced by peripheral nerve injury or sciatic inflammatory neuritis. This effect is associated with a reduction in pro-inflammatory cytokines, as well as a reduction in local leukocyte number and their capacity to produce pro-inflammatory cytokines. Curiously, clonidine's antinociceptive effect appears with a 2-3-day delay after injection. Previous observations have shown that alpha-adrenoceptor activation induces apoptosis in leukocytes, which would reduce leukocyte number. Additionally, macrophage scavenging of apoptotic cells results in a shift to an anti-inflammatory phenotype, with expression of transforming growth factor (TGF) beta1. We therefore examined the effects of perineural clonidine 24 h and 3 days after its injection on apoptosis, TGF-beta1 expression and lymphocyte and macrophage phenotype in acute sciatic inflammatory neuritis. Perineural clonidine reduced ipsilateral neuritis-induced hypersensitivity in a delayed manner (3 days after treatment), along with a reduction at this time in lymphocyte number and an increase in caspase-3 and TGF-beta1 expressing cells and macrophages co expressing TGF-beta1 in the sciatic nerve. One day after injection clonidine treatment was associated with a reduction in lymphocytes and pro-inflammatory Th 1 cells as well as increased numbers of caspase-3 and TGF-beta1 expressing cells and macrophages co-expressing TGF-beta1 in sciatic nerve. Clonidine's effects were prevented by co-administration of an alpha2-adrenoceptor antagonist. These data suggest that alpha2-adrenoceptor activation in sciatic inflammatory neuritis increases local apoptosis and anti-inflammatory products early after treatment. This early effect likely underlies the delayed anti-inflammatory and anti hypersensitivity effects of perineural clonidine in this setting. PMID- 17079114 TI - Prenatal manganese levels linked to childhood behavioral disinhibition. AB - Although manganese (Mn) is an essential mineral, high concentrations of the metal can result in a neurotoxic syndrome affecting dopamine balance and behavior control. We report an exploratory study showing an association between Mn deposits in tooth enamel, dating to the 20th and 62-64th gestational weeks, and childhood behavioral outcomes. In a sample of 27 children, 20th week Mn level was significantly and positively correlated with measures of behavioral disinhibition, specifically, play with a forbidden toy (36 months), impulsive errors on a continuous performance and a children's Stroop test (54 months), parents' and teachers' ratings of externalizing and attention problems on the Child Behavior Checklist (1st and 3rd grades), and teacher ratings on the Disruptive Behavior Disorders Scale (3rd grade). By way of contrast, Mn level in tooth enamel formed at the 62-64th gestational week was correlated only with teachers' reports of externalizing behavior in 1st and 3rd grades. Although the source(s) of Mn exposure in this sample are unknown, one hypothesis, overabsorption of Mn secondary to gestational iron-deficiency anemia, is discussed. PMID- 17079115 TI - Femoral nerve palsy after mandibular reconstruction with microvascular iliac flap: a complication under anticoagulation therapy. AB - Pharmacological prophylaxis and intervention are used extensively in head and neck reconstructions with microvascular flaps. There is no universally accepted protocol, but the microvascular surgery literature recommends intraoperative anticoagulation with heparin. Here is reported a case of iliacus haematoma with subsequent femoral nerve palsy after the harvest of a microvascular iliac flap for mandibular reconstruction in a patient who had been treated with heparin. The association between femoral nerve palsy and anticoagulant therapy has been well described. It remains unclear as to why the iliacus muscle is particularly vulnerable to intramuscular haemorrhage. Femoral nerve neuropathy is also an uncommon but recognized complication after abdominopelvic surgery. Iliacus haematoma secondary to microvascular surgery has not been previously reported. This case illustrates the need to be aware of this type of complication and its clinical manifestations for patients under anticoagulant therapy in the perioperative period during microvascular reconstructions. PMID- 17079116 TI - More severe epilepsy and cognitive impairment in the offspring of a mother with mosaicism for the ring 20 chromosome. AB - PURPOSE: Ring chromosome 20 [r(20)] syndrome is a rare chromosomal disorder. Cases tend to be sporadic. We elucidate the characteristics of an inherited r(20) mosaicism by describing the clinical features of three family members: a mother and her two children. RESULTS: The mosaicism rate of the mother was 10% and that of the children 40%. The mother experienced her first epileptic seizures at 24 years of age. Epilepsy was diagnosed two years later. After an unstable period lasting 3 years, she has been seizure-free for 13 years on a combination of valproate and lamotrigine. She has normal intelligence with full working capacity. The daughter exhibited her first epileptic seizures at the age of 7 years and she continues to have seizures weekly. The first epileptic seizures in the son were observed at 5 years of age. The son's epilepsy has been drug resistant from the onset, and a vagal nerve stimulator (VNS) has been ineffective. Psychomotor development was normal in both children up to the onset of epilepsy. Learning difficulties increased throughout school age and both children needed special educational programs. Neuropsychological evaluations have shown deterioration of cognitive levels. Both children had behavioural problems during school age but no longer in adolescence. All three subjects are nondysmophic, normocephalic and of normal growth. CONCLUSION: In this family the phenotype of r(20) mosaicism seems to be more severe in the successive generation along with a greater level of mosaicism. The aggravated clinical picture in inherited r(20) mosaicism concerned the onset of epilepsy, drug responsiveness, the cognitive level and behavioural features. PMID- 17079118 TI - No association between the Nuclear Receptor Coactivator 7 gene and schizophrenia in the Chinese Han population. PMID- 17079117 TI - Minor physical anomalies in schizophrenia: a meta-analysis. AB - Numerous studies report an increased frequency of minor physical anomalies (MPAs) in schizophrenic individuals compared with controls. However, these studies vary considerably regarding the magnitude of the case-control disparity and the topographical distribution of the anomalies. A meta-analysis was carried out on the existing MPA literature in an effort to better understand the relationship between MPAs and schizophrenia. Following a literature search, 13 studies were identified that met our inclusion criteria. Mean total MPA scores were available for 11 of these studies, whereas only seven studies provided regional MPA scores. For both the total MPA and regional MPA analyses, pooled effect sizes (Hedges' g and pooled odds ratios, respectively) were calculated along with tests of heterogeneity. For the total MPA analyses, a meta-regression approach was used to explore the relationship between possible moderator variables (e.g., number of MPA scale items) and effect size heterogeneity. The magnitude of the pooled effect size for the total MPA scores was high (1.131; p<0.001), indicating significantly more overall MPAs in schizophrenic individuals. Significant effect size heterogeneity was present (p<0.001); however, this heterogeneity could not be explained by any of the included moderator variables. The regional MPA analysis revealed significantly increased MPAs in all six anatomical regions (p<0.05), although the pooled odds ratios for these regions did not differ significantly from one another. These results suggest a lack of regional specificity for MPAs in schizophrenia. PMID- 17079119 TI - Neurosteroids in child and adolescent psychopathology. AB - Neurosteroids play a significant role in neurodevelopment and are involved in a wide variety of psychopathological processes. There is accumulating evidence on their role in adult psychopathology, including Alzheimer disease, schizophrenia, mood disorder, anxiety disorders and post-traumatic stress disorder. Little is known, however, about the possible role of neurosteroids in child and adolescent psychopathology although there is increasing evidence for their critical role from the early stages of brain development until adolescence. In this review we focus on the involvement of neurosteroids in neurodevelopment and mental disorders in children and adolescents. Adequate physiological levels protect the developing neural system from insult and contribute to the regulation of brain organization and function. Neurosteroids may be involved in the pathophysiology and pharmacotherapy of a variety of disorders in children and adolescents, including schizophrenia, depression, eating disorders, aggressive behavior and attention deficit. The complex interaction between neurosteroids, neurodevelopment, life-events, genetics and mental disorders in children and adolescents merits further investigation. PMID- 17079120 TI - Synthesis of indolizine derivatives with selective antibacterial activity against Mycobacterium tuberculosis. AB - 1-substituted indolizines with activity against Mycobacterium tuberculosis have been synthesized. The most active compounds carry an hydroxyphenylmethyl- or hydroxyalkyl substituent in the indolizine 1-position. The alkyl chain should be moderately long (C-5 or C-6). Aryl groups in the 2- and 3-position of the indolizine are also required. Removal of the 3-substituent resulted in significant loss of activity. A nitrile substituent in the 7-position is beneficial for both chemical stability and bioactivity. The compounds studied display a narrow antibacterial spectrum and appear to be quite selective antimycobacterial compounds. Moderate activity against certain pathogenic protozoa was also observed. PMID- 17079122 TI - From Beijing 2006 towards Montreal ISP2010. PMID- 17079123 TI - [Omega 3: is there a situation of deficiency in young children?]. AB - This paper starts with a review of the metabolism of n-6 (omega6) and n-3 (omega3) fatty acids, the resulting eicosanoids (prostaglandins, leucotrienes, and thromboxanes), and the physiological functions they are involved in, with special emphasis on effects during pregnancy (such as possible benefits on fetal growth, prevention of hypertension of pregnancy, and prevention of premature labor). Attention is then turned to the key role for long-chain polyunsaturated fatty acids (LCPUFAs), most notably docosahexaenoic acid (DHA), in central nervous system and retinal cell membrane structure and in cerebral and retinal development. Massive maternofetal transfer of LCPUFAs occurs during the third trimester of pregnancy, so that maintaining an adequate intake of DHA during pregnancy is crucial. Preterm babies must receive sufficient amounts of DHA, either via breast milk or via formula supplemented with LCPUFAs, both of which prevent DHA levels from declining in blood and cerebral phospholipids. These two methods of achieving an adequate DHA intake ensure normal maturation of visual acuity and cognitive function, as shown by randomized controlled trials. Formula supplemented with fish oils rich in n-3 LCPUFAs but lacking a proportionate supply of arachidonic acid (ArA, 20: 4n-6) negatively affects somatic growth, confirming the need for an adequate ArA supply. Eicosapentaenoic acid (EPA) can also exert negative effects. Therefore, the best source of supplemental LCPUFAs may be oil from single-cell algae and microscopic fungi, which contains adequate amounts of DHA and ArA. In full term neonates, strong arguments support LCPUFA supplementation, despite continuing controversy generated by conflicting results from interventional studies. These discrepancies in study results may be ascribable to differences in study design, patient age, intervention duration, assessment tool sensitivity, and LCPUFA sources. In 1996, an amendment to the European Directive on infant formulas and follow-on formulas was developed to authorize LCPUFA supplementation and to specify appropriate ranges. As a result, formulas supplemented with DHA and ArA were introduced on the market. Pregnant and nursing women should be advised to maintain an adequate dietary intake of DHA in order to meet their increased needs and those of the fetus or infant. PMID- 17079121 TI - Experimental-neuromodeling framework for understanding auditory object processing: integrating data across multiple scales. AB - In this article, we review a combined experimental-neuromodeling framework for understanding brain function with a specific application to auditory object processing. Within this framework, a model is constructed using the best available experimental data and is used to make predictions. The predictions are verified by conducting specific or directed experiments and the resulting data are matched with the simulated data. The model is refined or tested on new data and generates new predictions. The predictions in turn lead to better-focused experiments. The auditory object processing model was constructed using available neurophysiological and neuroanatomical data from mammalian studies of auditory object processing in the cortex. Auditory objects are brief sounds such as syllables, words, melodic fragments, etc. The model can simultaneously simulate neuronal activity at a columnar level and neuroimaging activity at a systems level while processing frequency-modulated tones in a delayed-match-to-sample task. The simulated neuroimaging activity was quantitatively matched with neuroimaging data obtained from experiments; both the simulations and the experiments used similar tasks, sounds, and other experimental parameters. We then used the model to investigate the neural bases of the auditory continuity illusion, a type of perceptual grouping phenomenon, without changing any of its parameters. Perceptual grouping enables the auditory system to integrate brief, disparate sounds into cohesive perceptual units. The neural mechanisms underlying auditory continuity illusion have not been studied extensively with conventional neuroimaging or electrophysiological techniques. Our modeling results agree with behavioral studies in humans and an electrophysiological study in cats. The results predict a particular set of bottom-up cortical processing mechanisms that implement perceptual grouping, and also attest to the robustness of our model. PMID- 17079124 TI - [Early bacterial colonisation of the intestine: why it matters?]. AB - The birth process allows the progressive formation of complex intestinal microflora composed of myriad bacteria, leading to this recently identified host bacterial mutualism in the human intestine. This kind of cross-talk originating from birth is opportunistically used by the young host to initiate its own immune system. Recent epidemiogical data support the hypothesis that some increasing immune deviances observed in the last 2 decades could have originated from a modification of the bacterial environment in young populations. Our modern approach to perinatal care may, to some extent, have modified inadequately the overall quality of this bacterial-host interface. The international medical community has to be made aware of the increasing importance that initial colonising intestinal microflora could have on the health and well-being of the host later in life. It is of great concern to decrease these possible negative influences and to discover in the near future the possible means of helping to manipulate positively the gut microbiotia of infants. PMID- 17079125 TI - Asthma control in Spain. Do season and treatment pattern matter? The ESCASE study. AB - The aim of this study was to assess the degree of asthma control according to GINA criteria during two different seasons in Spain. An multicenter, longitudinal, epidemiological study with the participation of a sample of physicians in Spain was conducted. Consecutive asthma patients, 18 years of age and older, seeking primary and specialist care were included in the study. Patients were seen during the winter and spring 2004 and were asked about asthma control according to GINA control criteria (daytime and nighttime symptoms, asthma exacerbations, limitations of physical activity, and visits to the emergency department) during the 4 weeks prior to the visit. Control was defined according to daytime and nighttime symptoms. A total of 614 patients participated in the study. The proportion of patients reporting daytime symptoms "every day" or "most days" during the winter versus spring was 40.1% vs. 23% (P<0.01); 26.9% vs. 14.1% presented symptoms at night (P<0.01); 11.5% vs. 8.3% had severe exacerbations; 33.5% vs. 35.7% presented symptoms accompanying exercise, and 9.4% vs. 4.3% (P<0.01) had required emergency visits. The number of patients with inadequate control was slightly higher in winter than in spring (74.4% vs. 71%) (P<0.01). The most commonly prescribed treatment was ICS plus LABAs for both periods. Asthma is poorly controlled in Spain and strategies are needed to improve management of this illness. PMID- 17079126 TI - Digestible and indigestible carbohydrates: interactions with postprandial lipid metabolism. AB - The balance between fats and carbohydrates in the human diet is still a matter of very active debate. Indeed, the processing of ordinary mixed meals involves complex processes within the lumen of the upper digestive tract for digestion, in the small intestine mucosa for absorption and resecretion, and in peripheral tissues and in the circulation for final handling. The purpose of this review is to focus on available knowledge on the interactions of digestible or indigestible carbohydrates with lipid and lipoprotein metabolism in the postprandial state. The observations made in humans after test meals are reported and interpreted in the light of recent findings on the cellular and molecular levels regarding possible interplays between carbohydrates and lipid moieties in some metabolic pathways. Digestible carbohydrates, especially readily digestible starches or fructose, have been shown to exacerbate and/or delay postprandial lipemia, whereas some fiber sources can lower it. While interactions between dietary fibers and the process of lipid digestion and absorption have been studied mainly in the last decades, recent studies have shown that dietary carbohydrate moieties (e.g., glucose) can stimulate the intestinal uptake of cholesterol and lipid resecretion. In addition to the well-known glucose/fructose transporters, a number of transport proteins have recently been involved in intestinal lipid processing, whose implications in such interactions are discussed. The potential importance of postprandial insulinemia in these processes is also evaluated in the light of recent findings. The interactions of carbohydrates and lipid moieties in the postprandial state may result from both acute and chronic effects, both at transcriptional and posttranscriptional levels. PMID- 17079127 TI - Rapid method for detection of Salmonella in milk by surface plasmon resonance (SPR). AB - The Plasmonic surface plasmon resonance (SPR) device was used to develop a rapid, simple and specific immunoassay for detection of Salmonella in milk. Rapid detection of Salmonella contamination is a major challenge for the food industry. Salmonella contamination is well known in all foods including pasteurised milk. The SPR assay was developed as a sandwich model using a polyclonal antibody against Salmonella as capture and detection antibody. Milk spiked with Salmonella typhimurium cells, killed by thimerosal (1%, w/w) treatment was used. Using the Plasmonic SPR assay it was possible to detect S. typhimurium down to a concentration of 1.25 x 10(5) cells ml(-1) in both milk and buffer system. The results obtained are comparable with existing, approved rapid Salmonella detection techniques. No negative effects on the sensitivity of the assay are encountered due to the milk matrix. Hence, no sample preparation or clean-up steps are required. The sample volume requirement for the assay is only 10 microl. Using the assay S. typhimurium was detected in milk within 1h, whereas the cultural techniques require 3-4 days for presumptive positive isolates and further time for confirmation. The rapid tests require at least 24h for the results. The Plasmonic SPR device operates on the Kretschmann configuration and is a cuvette-based system with the advantage of having eight channels on one single SPR chip. PMID- 17079128 TI - Analysis of proteolytic degradation of a crude protein mixture using a surface acoustic wave sensor. AB - Degradation of a crude protein mixture by proteases with pH optima from acidic to basic was followed in real time using a surface acoustic wave biosensor in Love wave geometry. Proteases EC 3.4.23.18 from Aspergillus saitoi, EC 3.4.21.62 from Bacillus licheniformis, and Novozyme from Bacillus sp. have been used. Kinetic constants extracted from the curves resulted for comparable protease concentrations in maximal degradation rates between 1.1 x 10(-2) and 1.5 x 10( 2)s(-1). For the three proteases investigated, similar amounts of up to about two thirds of the estimated 28 ng/cm2 bound molecules were proteolyzed. The residual material not degraded by the proteases was removed from the surface with 0.5% SDS. The analysis of the sensor signal allows: (1) estimation of the total mass of protein bound to the sensor surface and of the degradable fraction; (2) extraction of the pure mass signal; and (3) kinetic evaluation. PMID- 17079129 TI - An experimental and theoretical study of the morphine binding capacity and kinetics of an engineered opioid receptor. AB - Electrochemical real-time monitoring of ligand binding to an engineered opioid receptor specific for morphine is reported. In the particular systems studied, 90% of the binding was found to be completed after only 85-120 s. Thus, the binding kinetics has proven to be more rapid than previously believed. The observed association rate constant for the morphine binding reaction was calculated to be 215 M(-1)s(-1). A theoretical analysis of the experimental binding data suggested that the binding sites of the engineered opioid receptor could best be described by a model having two populations of binding sites: K(D)=40 microM (13 micromol/g) and K(D)=205 microM (29 micromol/g). Furthermore, a theoretical model was developed in order to explain the observed binding of the engineered opioid receptor. This model suggested that the binding sites on the polymer surface are up to 5.1A deep and they allow 100% of the ligand (morphine) to anchor itself into the site. The predicted theoretical maximum binding capacity for the reported receptor is calculated to be approximately 2 mmol/g polymer (based on an increase of cavity density). PMID- 17079130 TI - Protein-protein interactions in the allosteric regulation of protein kinases. AB - Protein-protein interactions involving the catalytic domain of protein kinases are likely to be generally important in the regulation of signal transduction pathways, but are rather sparsely represented in crystal structures. Recently determined structures of the kinase domains of the mitogen-activated protein kinase Fus3, the RNA-dependent kinase PKR, the epidermal growth factor receptor and Ca(2+)/calmodulin-dependent protein kinase II have revealed unexpected and distinct mechanisms by which interactions with the catalytic domain can modulate kinase activity. PMID- 17079131 TI - Molecular gymnastics: serpin structure, folding and misfolding. AB - The native state of serpins represents a long-lived intermediate or metastable structure on the serpin folding pathway. Upon interaction with a protease, the serpin trap is sprung and the molecule continues to fold into a more stable conformation. However, thermodynamic stability can also be achieved through alternative, unproductive folding pathways that result in the formation of inactive conformations. Our increasing understanding of the mechanism of protease inhibition and the dynamics of native serpin structures has begun to reveal how evolution has harnessed the actual process of protein folding (rather than the final folded outcome) to elegantly achieve function. The cost of using metastability for function, however, is an increased propensity for misfolding. PMID- 17079132 TI - Cut and move: protein machinery for DNA processing in bacterial conjugation. AB - Conjugation is a paradigmatic example of horizontal or lateral gene transfer, whereby DNA is translocated between bacterial cells. It provides a route for the rapid acquisition of new genetic information. Increased antibiotic resistance among pathogens is a troubling consequence of this microbial capacity. DNA transfer across cell membranes requires a sophisticated molecular machinery that involves the participation of several proteins in DNA processing and replication, cell recruitment, and the transport of DNA and proteins from donor to recipient cells. Although bacterial conjugation was first reported in the 1940s, only now are we beginning to unravel the molecular mechanisms behind this process. In particular, structural biology is revealing the detailed molecular architecture of several of the pieces involved. PMID- 17079133 TI - Docking interactions in protein kinase and phosphatase networks. AB - To achieve high biological specificity, protein kinases and phosphatases often recognize their targets through interactions that occur outside of the active site. Although the role of modular protein-protein interaction domains in kinase and phosphatase signaling has been well characterized, it is becoming clear that many kinases and phosphatases utilize docking interactions - recognition of a short peptide motif in target partners by a groove on the catalytic domain that is separate from the active site. Docking is particularly prevalent in serine/threonine kinases and phosphatases, and is a versatile organizational tool for building complex signaling networks; it confers a high degree of specificity and, in some cases, allosteric regulation. PMID- 17079134 TI - Identification of differentially expressed genes in HPV-positive and HPV-negative oropharyngeal squamous cell carcinomas. AB - Human papillomaviruses (HPVs) have been implicated in the pathogenesis of a subset of squamous cell carcinoma of the head and neck (SCCHN). The goal of this study was to compare the cellular gene expression profiles of HPV-positive and HPV-negative oropharyngeal carcinomas with those of the normal oral epithelium. Using Affymetrix Human U133A GeneChip, our results showed that 397 genes were differentially expressed in HPV-positive SCCHN compared to the normal oral epithelium. The upregulated genes included those involved in cell cycle regulation (CDKN2A), cell differentiation (SFRP4) and DNA repair (RAD51AP1), while the downregulated genes included those involved in proteolysis (PRSS3). We also found 162 differentially expressed genes in HPV-negative SCCHN compared to the normal oral mucosa. The upregulated genes included those involved in cell proliferation (AKR1C3) and transcription regulation (SNAPC1), while downregulated genes included those involved in apoptosis (CLU) and RNA processing (RBM3). Our studies also identified a subgroup of 59 differentially expressed genes in HPV positive SCCHN as compared to both HPV-negative SCCHN and normal oral tissues. Such upregulated genes included those involved in nuclear structure and meiosis (SYCP2), DNA repair (RFC5), and transcription regulation (ZNF238). Genes involved in proteolysis (KLK8) and signal transduction (CRABP2) were found to be downregulated in HPV-positive SCCHN. The results of GeneChip experiments were validated by quantitative real-time RT-PCR analysis of a few representative genes. Our results reveal specific gene expression patterns in HPV-positive and HPV-negative oropharyngeal squamous carcinomas that may serve as potential biomarkers for the development of SCCHN. PMID- 17079135 TI - Mapping the emergence and development of translational cancer research. AB - Cancer research is one of the principal targets of translational research, yet the nature of the relationships between different forms of cancer research remains controversial. The paper examines publications in the cancer field during the 1980-2000 period. A network analysis software program was used to map evolving patterns of inter-citations between cancer publications, their different research levels and the transformation of their relational content. Both inter citation and content maps provide striking evidence of the consolidation in the 1990s of a translational interface that was practically non existent a few decades before. In 1980, research was polarized according to the allegiance to either a clinical or a laboratory style. This same duality obtains in the year 2000, albeit with the additional presence of a third, biomedical player whose activities are similarly structured by a common orientation, rather than by an exclusive commitment to a specific sub-domain. PMID- 17079136 TI - Gastrointestinal metastases from primary lung cancer. AB - The aim of this study was to evaluate gastrointestinal metastases from primary lung cancer confirmed by autopsy. We identified and examined patients with a diagnosis of primary lung cancer over 33 years. We also reviewed patients with gastrointestinal metastases including the stomach, small bowel, and large bowel. This study comprised 470 patients with lung cancer. We detected 56 (11.9%) cases with gastrointestinal metastases. There were 12 (30%) cases with gastrointestinal metastases among 40 cases with large cell carcinoma. The histological type of large cell carcinoma led to a significantly higher rate of gastrointestinal metastases compared with that of non-large cell carcinoma (P=0.004, odds ratio 3.524). Life threatening gastrointestinal metastases occurred in 12 cases and five occurred in large cell carcinoma. Gastrointestinal metastases from primary lung cancer may occur in the clinical course and result in life threatening gastrointestinal metastases, particularly if patients have the histological type of large cell carcinoma. PMID- 17079137 TI - Regulation of Vitamin D hydroxylases gene expression by 1,25-dihydroxyvitamin D3 and cyclic AMP in cultured human syncytiotrophoblasts. AB - Human placenta synthesizes and metabolizes 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)/calcitriol] through the activity of 25-hydroxyvitamin D(3) 1alpha-hydroxylase (CYP27B1) and 1,25(OH)(2)D(3)-24-hydroxylase (CYP24A1), the two key enzymes for Vitamin D metabolism. In this study, calcitriol rapidly generated intracellular cAMP accumulation in cultured human syncytiotrophoblast cells, which in turn enhanced hCG secretion, a marker of trophoblast endocrine activity. The effects of 1,25(OH)(2)D(3) upon the expression of CYP27B1 and CYP24A1 were also investigated. 1,25(OH)(2)D(3) and activators of the PKA signaling system decreased the expression of CYP27B1, whereas increased CYP24A1 gene transcription. The use of a selective inhibitor of PKA (H-89) prevented the effects of calcitriol on CYP27B1 gene and hCG secretion, but not on CYP24A1 transcription. Addition of ZK 159222, a Vitamin D receptor (VDR) antagonist, blocked the calcitriol-mediated upregulation of 24-hydroxylase gene expression but did not affect calcitriol-induced downregulation of CYP27B1 gene or hCG stimulation. In addition, our study also demonstrated a role of calcitonin on Vitamin D hydroxylases gene regulation in placenta. The overall data suggest that calcitriol downregulates CYP27B1 expression via a cAMP-dependent signaling pathway, whereas upregulates 24-hydroxylase gene expression through a VDR dependent mechanism. PMID- 17079138 TI - 0.2kb promoter sequence of the murine Cyp19 gene target beta-galactosidase expression to specific brain areas of transgenic mice. AB - Cyp19 encodes the key enzyme of estrogen biosynthesis, aromatase cytochrome P450. In mice it is mainly expressed in the ovary and brain, where transcription is directed by a distal, brain-specific promoter (P(br)). In order to map functional sequence elements of P(br), portions of various length (0.2, 1.0, and 1.7[kb]) were fused to a lacZ reporter gene and analyzed in transgenic mice. Numbers of integrated reporter genes varied from 1 to 23 copies in different transgenic lines. These copy numbers however did not show any correlation to the levels of transgene expression. All of the constructs were found being expressed in the olfactory bulb, limbic cortex, amygdala, and hypothalamus. Additional expression in thalamic nuclei, bed nucleus of stria terminalis, and dorsal mesencephalon was found in transgenic lines with constructs 1.0 and 1.7, and expression in septal and preoptic nuclei was only found with construct 1.7. The data demonstrate that 0.2kb of P(br) target reporter gene expression to specific brain areas. The data also strongly suggest that the sequence between 0.2 and 1.7kb upstream, is necessary for expression in additional areas. However even 1.7kb of P(br) are not sufficient to consistently mimic the accurate expression pattern of Cyp19. PMID- 17079140 TI - Serendipitous discovery of novel imidazolopyrazole scaffold as selective androgen receptor modulators. AB - A novel imidazolopyrazole derivative has been fortuitously discovered as potent selective androgen receptor modulator with in vivo efficacy. PMID- 17079139 TI - Seed vigour studies in corn, soybean and tomato in response to fish protein hydrolysates and consequences on phenolic-linked responses. AB - Seed priming with fish protein hydrolysates (FPH) has been studied for the enhancement of seed vigour of corn, soybean and tomato. The influence of FPH at 2.5 mL/L and 5.0 mL/L on traditional agronomic parameters for seed vigour (germination percentage, seedling weight, seedling height) and potential new vigour-associated parameters (phenolic content, antioxidant activity, guaiacol peroxidase (GuPX) activity, chlorophyll content) was investigated. FPH treatment preferentially stimulated seedling vigour in the following order: soybean>tomato>corn. For soybean, FPH at 2.5 mL/L and 5 mL/L improved the majority of the seed vigour parameters (seedling weight and height, phenolic content, antioxidant activity and chlorophyll content, and lignification associated GuPX activity). Similarly, for tomato, FPH at 2.5 mL/L stimulated seedling weight and height, GuPX activity and chlorophyll content. However, FPH did not stimulate corn seed vigour. Our results suggest an ability of proline precursor-rich FPH to improve of plant growth and development (e.g., seed vigour) in phenolic-rich plant species through modulation of phenolic and chlorophyll metabolisms. PMID- 17079141 TI - Discovery of gamma-secretase inhibitors efficacious in a transgenic animal model of Alzheimer's disease. AB - Attachment of the cyclopropylcarbamate group to the piperidine core of gamma secretase inhibitors leads to a dramatic increase of their in vitro potency. Strategies for subsequent improvement of the in vivo pharmacokinetic profile of the series are discussed. Resulting compounds significantly reduce Abeta levels in TgCRND8 mice after a single PO dosing at 30 mpk. PMID- 17079142 TI - 5-HT2C antagonists based on fused heterotricyclic templates: design, synthesis and biological evaluation. AB - Design, synthesis and properties of a new tricyclic series of selective 5-HT2C receptor antagonists are reported. Conformational analysis of a 2-phenyl dihydropyrrolone scaffold suggested that ring fusion, locking coplanarity between the rings of this moiety, might be tolerated by the 5-HT2C receptor. An interesting effect of this is the change of the nature of the carbon-carbon double bond of the lactam ring from vinylic to aromatic. The changes were found to result in a favourable profile at both, receptor and in vivo level. PMID- 17079143 TI - Development of a novel therapeutic suppressor of brain proinflammatory cytokine up-regulation that attenuates synaptic dysfunction and behavioral deficits. AB - We report the development of a novel, aqueous-soluble, safe, small molecule, experimental therapeutic that suppresses injury-induced, proinflammatory cytokine increases in the brain, with resultant attenuation of synaptic protein biomarker loss and improvement in hippocampus-dependent behavioral deficits. A GMP production scheme for the active pharmaceutical ingredient, compound 17, is presented. The development and large-scale availability of this novel compound allow exploration of new, potentially disease-modifying, therapeutic approaches to CNS disorders. PMID- 17079144 TI - Novel nucleotide triphosphates as potent P2Y2 agonists with enhanced stability over UTP. AB - The synthesis of a series of novel C-linked nucleotide triphosphates is reported. These exhibit excellent agonist potency and selectivity for the P2Y2 receptor with a number of examples having EC50 values below 10 nM. Representative compounds from the N-linked and C-linked series showed enhanced metabolic stability compared with that of the natural ligand UTP. PMID- 17079145 TI - Agreement between footswitch and ground reaction force techniques for identifying gait events: inter-session repeatability and the effect of walking speed. AB - This study assessed the inter-session repeatability of agreement between foot ground contact events (foot contact and foot off) identified from ground reaction force and footswitch-based techniques over a range of gait speeds. No inter session differences in agreement between the ground reaction force and footswitch based techniques were identified and while agreement between foot-ground contact events defined by the two techniques was found to differ with walking speed, the differences were small (<6 ms or approximately 1% stance duration) and therefore of little practical significance to the majority of gait studies. PMID- 17079147 TI - Synthesis of a non-cationic, water-soluble perylenetetracarboxylic diimide and its interactions with G-quadruplex-forming DNA. AB - A number of N,N'-disubstituted perylenetetracarboxylic diimides have been reported to bind effectively to DNA that adopts G-quadruplex motifs. In some cases, this binding may actively drive the transition from single-strand DNA to the quadruplex form. The perylenediimides in the reported cases all have amine containing side chains, which are thought to interact with the grooves of the quadruplex and help dictate the selectivity of these compounds for quadruplex versus duplex DNA. We synthesized a polyethyleneglycol-swallowtailed (PEG-tailed) perylenediimide that is water-soluble even though it is uncharged. Binding to duplex and quadruplex DNA of this perylenediimide was studied by fluorescence quenching titrations under a variety of salt conditions, and the compound's effect on quadruplex formation was studied by non-denaturing gel electrophoresis. Our results indicate that while the molecule binds to single-stranded DNA quite effectively and with selectivity, it does not drive the transition of the DNA to the tetrameric quadruplex structure, supporting the idea that charge neutralization is a key component of perylene compounds that stabilize tetrameric quadruplexes. PMID- 17079146 TI - Roles of phosphatidate phosphatase enzymes in lipid metabolism. AB - Phosphatidate phosphatase (PAP) enzymes catalyze the dephosphorylation of phosphatidate, yielding diacylglycerol and inorganic phosphate. In eukaryotic cells, PAP activity has a central role in the synthesis of phospholipids and triacylglycerol through its product diacylglycerol, and it also generates and/or degrades lipid-signaling molecules that are related to phosphatidate. There are two types of PAP enzyme, Mg(2+) dependent (PAP1) and Mg(2+) independent (PAP2), but only genes encoding PAP2 enzymes had been identified until recently, when a gene (PAH1) encoding a PAP1 enzyme was found in Saccharomyces cerevisiae. This discovery has revealed a molecular function of the mammalian protein lipin, a deficiency of which causes lipodystrophy in mice. With molecular information now available for both types of PAP, the specific roles of these enzymes in lipid metabolism are being clarified. PMID- 17079148 TI - Synthesis and pharmacological evaluation of some 3-phenyl-2-substituted-3H quinazolin-4-one as analgesic, anti-inflammatory agents. AB - A variety of novel 3-phenyl-2-substituted-3H-quinazolin-4-ones were synthesized by reacting the amino group of 2-hydrazino-3-phenyl-3H-quinazolin-4-one with different aldehydes and ketones. The starting material 2-hydrazino-3-phenyl-3H quinazolin-4-one was synthesized from aniline. The title compounds were investigated for analgesic, anti-inflammatory and ulcerogenic index activities. While the test compounds exhibited significant activity, compounds, 2-(N'-2 butylidene-hydrazino)-3-phenyl-3H-quinazolin-4-one (AS1), 2-(N'-3-pentylidene hydrazino)-3-phenyl-3H-quinazolin-4-one (AS2) and 2-(N'-2-pentylidene-hydrazino) 3-phenyl-3H-quinazolin-4-one (AS3), exhibited moderate analgesic activity. The compound 2-(N'-2-pentylidene-hydrazino)-3-phenyl-3H-quinazolin-4-one (AS3) showed more potent anti-inflammatory activity when compared to the reference standard diclofenac sodium. Interestingly, the test compounds showed only mild ulcerogenic side effect when compared to aspirin. PMID- 17079149 TI - Fluoro-ketopyranosyl nucleosides: synthesis and biological evaluation of 3-fluoro 2-keto-beta-D-glucopyranosyl derivatives of N4-benzoyl cytosine. AB - 1,2:5,6-Di-O-isopropylidene-alpha-d-glucofuranose on mild oxidation, reduction, fluorination, and deisopropylidenation followed by acetylation gave peracetylated 3-deoxy-3-fluoro-d-glucopyranose. This was coupled with silylated N(4)-benzoyl cytosine. The nucleoside was deacetylated and after several subsequent protection and deprotection steps afforded the desired 3-fluoro-2-keto-beta-d-glucopyranosyl derivatives. These novel synthesized compounds were evaluated for antiviral and cytotoxic activities against rotavirus, vesicular stomatitis virus, and the human colon adenocarcinoma cell line Caco-2, and have a promising potential in combating the rotaviral infections and in the treatment of colon cancer. As compared to AZT, a nucleoside analogue of reverse transcriptase inhibitor, the novel synthesized 1-(3,4-dideoxy-3-fluoro-beta-d-glycero-hex-3-enopyranosyl-2 ulose)-N(4)-benzoyl cytosine showed to be more effective at lower concentrations in inhibition of rotavirus infection as well as in the same range of antitumor activity. PMID- 17079151 TI - Antinociceptive and antiinflammatory activities and QSAR studies on 2-substituted 4,5-diphenyl-1H-imidazoles. AB - This paper describes the pharmacological evaluation pertaining to antinociceptive (hot plate and tail flick) and antiinflammatory (based on Carrageenan-induced paw oedema) activities, and QSAR studies on 2-substituted-4,5-diphenyl-1H-imidazoles. Compounds with phenyl substitution with -F, -Cl, -NH(2), -N(CH(3))(2), -OH and OCH(3) at the p-position showed higher activity than the other substitutions in all the three studies. QSARs developed for the 60 and 120s hot plate data indicate that the models for both the cases not only fit the data very well (R(2)>0.9, R(adj)(2)>0.86), but also have very good predictive capability (q(2)>0.81). The descriptors used in the model relate to surface area, volume, dipole moment and ADME properties of the molecule. Good QSARs for the 60 and 120s tail flick data are developed. The models fit the data well (R(2)>0.8, R(adj)(2)>0.74), and in addition have good predictive capability (q(2)>0.66). Surface area, specifically polar surface area, HOMO and molecular connectivity index appear in the models. Very good QSAR model is developed for the antiinflammatory data (R(2)=0.86, R(adj)(2)=0.822 and q(2)=0.64) with aqueous solubility, number of hydrogen bond donor groups, surface area and principal moment of inertia as the molecular descriptors. PMID- 17079150 TI - Analgesic agents without gastric damage: design and synthesis of structurally simple benzenesulfonanilide-type cyclooxygenase-1-selective inhibitors. AB - In order to create novel analgesic agents without gastric disturbance, structurally simple cyclooxygenase-1 (COX-1) inhibitors with a benzenesulfonanilide skeleton were designed and synthesized. As a result, compounds 11f and 15a, which possess a p-amino group on the benzenesulfonyl moiety and p-chloro group on the anilino moiety, showed COX-1-selective inhibition. Moreover compound 11f, which is the most potent compound in this study showed more potent analgesic activity than that of aspirin at 30 mg/kg by po. The anti-inflammatory activity and gastric damage, however, were very weak or not detectably different from aspirin. Since the structure of our COX-1 inhibitors are very simple, they may be useful as lead compounds for superior COX 1 inhibitors as analgesic agents without gastric disturbance. PMID- 17079152 TI - Design of endoperoxides with anti-Candida activity. AB - Broad antifungal structure-activity relationships governing epoxy-endoperoxides 2 and 3 and their parent endoperoxides 1 are reported. Their inhibitory activity against Candida albicans in conjunction with hemolytic activity and/or growth inhibition of cultured mammalian cells are reported. This information provided guidance for the further development of endoperoxide and epoxy-endoperoxides as topical antifungal agents. PMID- 17079153 TI - Differential expression of litchi XET genes in relation to fruit growth. AB - Xyloglucan endotransglycosylase (XET) catalyses the transglycosylation of xyloglucan, the major hemicellulose polymer, which has been thought to mediate the cross-linking of cellulose microfibrils in cellular walls and proposed to be involved in the control of cell wall relaxation. To understand the relationship between litchi fruit cracking and gene expression patterns, three XET genes from litchi fruit were identified and then examined for their expression profiles in pericarp and aril tissues at different development stages, using a cracking resistant cultivar, 'Huaizhi', and a cracking-susceptible cultivar, 'Nuomici'. Three full-length cDNAs of 1267, 1095 and 1156 bp encoding XETs, named LcXET1, LcXET2 and LcXET3, respectively, were isolated from expanding fruit using RT-PCR and RACE-PCR (rapid amplification of cDNA ends) methods. Northern blotting analysis showed that LcXET1 mRNA accumulation occurred much earlier in aril tissues at 59 days after anthesis (DAA) than in pericarp tissues at 73 DAA in 'Nuomici'. However, it appeared at almost the same time (66 DAA) in pericarp and aril tissues in 'Huaizhi', which suggested that differential accumulation of LcXET1 in pericarp and aril tissues in 'Nuomici' and 'Huaizhi' was closely associated with fruit cracking. LcXET2 mRNA accumulation could be detected in pericarp and aril tissues throughout fruit development but exhibited a differential accumulation pattern between pericarp and aril tissues. In the aril of 'Nuomici', intensive signal bands were detectable at 59-73 DAA in rapidly expanding fruits of 'Nuomici' but only weak bands could be found in the pericarp tissues. In contrast, moderate signal bands were detectable both in pericarp and aril tissues of 'Huaizhi' fruits. Furthermore, LcXET3 showed constitutive expression in both pericarp and aril tissues of developing 'Nuomici' and 'Huaizhi' litchi fruit. In addition, differential expression patterns of three XETs genes were observed in different tissues of litchi, with only LcXET1 being fruit-specific. To further address the role of LcXET in fruit cracking, alpha naphthalene acetic acid (NAA) was used to treat 'Nuomoci' to reduce fruit cracking. Enhanced LcXET1 mRNA accumulation appeared in pericarp while LcXET2 and LcXET3 mRNA accumulation enhanced in aril tissues in the NAA-treated fruits. Thus, LcXET1 is more likely to play a role in reducing litchi fruit cracking than LcXET2 and LcXET3. PMID- 17079154 TI - Involvement of ethylene and lipid signalling in cadmium-induced programmed cell death in tomato suspension cells. AB - Cadmium-induced cell death was studied in suspension-cultured tomato (Lycopersicon esculentum Mill.) cells (line MsK8) treated with CdSO(4). Within 24 h, cadmium treatment induced cell death in a concentration-dependent manner. Cell cultures showed recovery after 2-3 days which indicates the existence of an adaptation mechanism. Cadmium-induced cell death was alleviated by the addition of sub muM concentrations of peptide inhibitors specific to human caspases indicating that cell death proceeds through a mechanism with similarities to animal programmed cell death (PCD, apoptosis). Cadmium-induced cell death was accompanied by an increased production of hydrogen peroxide (H(2)O(2)) and simultaneous addition of antioxidants greatly reduced cell death. Inhibitors of phospholipase C (PLC) and phospholipase D (PLD) signalling pathway intermediates reduced cadmium-induced cell death. Treatment with the G-protein activator mastoparan and a cell permeable analogue of the lipid signal second messenger phosphatidic acid (PA) induced cell death. Ethylene, while not inducing cell death when applied alone, stimulated cadmium-induced cell death. Application of the ethylene biosynthesis inhibitor aminoethoxy vinylglycine (AVG) reduced cadmium-induced cell death, and this effect was alleviated by simultaneous treatment with ethylene. Together the results show that cadmium induces PCD exhibiting apoptotic-like features. The cell death process requires increased H(2)O(2) production and activation of PLC, PLD and ethylene signalling pathways. PMID- 17079155 TI - In vitro evidence of Hsc70 functioning as a molecular chaperone during cold stress. AB - Hsp70 molecular chaperones have been shown to play an important role in helping cells to cope with adverse environments, especially in response to high temperatures. The molecular chaperone function of Hsc70 at low temperature was investigated. A cold-inducible spinach cytosolic Hsc70 was subcloned into a protein expression vector and the recombinant protein was expressed in bacterial cells. Recombinant Hsc70 bound a permanently unfolded substrate: alpha carboxymethylated lactalbumin (CMLA) in the presence of 3 mM ATP and MgCl(2) at low temperature (4 and -4 degrees C). Radiolabeling with (35)S-Met and (35)S-Cys and immunoprecipitation with cytosolic Hsc70 monoclonal antibodies showed that there were several proteins co-immunoprecipitated at low temperature (4 and -4 degrees C) but not at room temperature. Enhanced co-purification of sHsp17.7 with Hsc70 at low temperature was observed and suggests that co-chaperone interactions can contribute to molecular chaperone function during cold stress. These results suggest that the molecular chaperone Hsc70 may have a functional role in plants during low temperature stress. PMID- 17079156 TI - The evidence on which to base practice: Different tools for different times. PMID- 17079158 TI - Increased adiponectin is negatively linked to the local inflammatory process in patients with rheumatoid arthritis. AB - Adiponectin has been shown to exert insulin-sensitizing, anti-atherogenic, and anti-inflammatory properties in metabolic diseases. It has been suggested that adiponectin may play a role in rheumatoid arthritis (RA). To assess adiponectin in serum and synovial fluid from patients with RA and osteoarthritis (OA), and in serum from healthy controls. Adiponectin and CRP levels were analyzed by ELISA. The clinical activity of RA patients was assessed according to the 28 joint count Disease Activity Score. Synovial fluid adiponectin was significantly higher in RA than in OA patients (p<0.001). Adiponectin was negatively associated with the leukocyte count in RA synovial fluid (r=-0.45, p<0.05). Serum adiponectin was higher in RA compared to healthy controls (p<0.02), however comparable to OA patients. Serum adiponectin was higher than in synovial fluid in both diseases (p<0.001). In general, women had higher adiponectin levels than men. Adiponectin was not related to age, disease duration, body mass index, or disease activity of RA patients. Adiponectin is decreased in synovial fluid compared to serum indicating that peripheral fat stores are major producers of adiponectin into the blood stream. However, increased synovial fluid adiponectin in RA patients may counterpart the local inflammatory process. PMID- 17079159 TI - Efficacy of DNA hypomethylating capacities of 5-aza-2'-deoxycytidine and its alpha anomer. AB - In this paper, we have compared hypomethylating ability of classical beta-d anomer of 5-aza-2'-deoxycytidine (5-aza-CdRf) and its alpha anomer in cell cultures. Alpha anomers of nucleosides generally exhibit low biological activity compared to their beta counterparts. It is reported that alpha anomer of 5-aza CdRf efficiently hypomethylated genomic DNA in human T-lymphoblastoid CCRF-CEM cells. Satellite 2 and 18S rDNA were hypomethylated by alpha anomer at concentrations comparable to the beta form. However, the toxicity of the alpha anomer was 4-fold less than that of beta form. Contrast to CCRF-CEM the A549 lung carcinoma cells, possessing negligible level of methylation at repetitive loci, were highly resistant to 5-aza-CdRf treatment suggesting that global genomic methylation might be needed to mediate cytotoxic effect of the drug. Possible mechanisms of inhibition of DNA methylation by alpha anomer are discussed. In conclusion, alpha anomer of 5-aza-CdRf displaying lower host cytotoxicity than the classical beta form may be of potential use in epigenetic therapy. PMID- 17079160 TI - Antiplatelet and antithrombotic activities of essential oil from wild Ocotea quixos (Lam.) Kosterm. (Lauraceae) calices from Amazonian Ecuador. AB - Ocotea quixos essential oil was shown to possess significant inhibitory activity of platelet aggregation and clot retraction in rodent plasma. This study is aimed at fully characterizing the antiplatelet activity of the whole essential oil and its main components trans-cinnamaldehyde and methyl cinnamate also in human plasma, at investigating the mechanism underlying such activity and at evaluating the potential antithrombotic activity of subacute treatment of mice with Ocotea essential oil. In vitro Ocotea essential oil and trans-cinnamaldehyde inhibited arachidonic acid-, U46619-, ADP-, phorbol12-myristate13-alcetate-, collagen induced platelet aggregation and thrombin-induced clot retraction in human and rodent plasma; Ocotea oil and trans-cinnamaldehyde competitively antagonized contractions induced by thromboxane A2 receptor agonist U46619 in rat isolated aortic ring (K(B) = 18 and 3.2 microg ml(-1), respectively). In vivo Ocotea oil, orally administered in a subacute treatment (30-100 mg kg(-1) day(-1) for 5 days) to mice, prevented acute thrombosis induced by collagen-epinephrine intravenous injection. This antithrombotic activity was not accompanied by pro-haemorragic side effect, as detected by the inactivity in bleeding test, thus showing a favourable safety profile compared to the conventional antiplatelet agent, acetylsalicylic acid. Present findings indicate that Ocotea essential oil possesses potent and safe antithrombotic activity attributable to its antiplatelet and vasorelaxant effects. The main constituent trans-cinnamaldehyde seems to be the primary responsible for this activity through a putative mechanism involving the inhibition of thromboxane A2 receptors. PMID- 17079161 TI - Alpha-methyl-5-HT, a 5-HT2 receptor agonist, stimulates beta2-adrenoceptors in guinea pig airway smooth muscle. AB - Alpha-methyl-5-HT is widely used as a high-affinity 5-HT(2) receptors agonist, though some studies have postulated that this drug also activates other serotonergic receptors. In the present work, we found that a wide range of concentrations of alpha-methyl-5-HT induced biphasic responses (contraction followed by relaxation) in guinea pig tracheal rings. The relaxing phase caused by 32microM alpha-methyl-5-HT was blocked by 0.1microM propranolol. Furthermore, during an ongoing histamine-induced contraction, alpha-methyl-5-HT (0.1 100microM) produced a concentration-dependent relaxation starting at 10microM. This relaxation was fully abolished by 0.1microM propranolol or 1microM ICI 118,551 (a selective beta(2)-adrenoceptor antagonist). Additionally, in electrophysiological recordings, 32microM alpha-methyl-5-HT also enhanced the membrane K(+) currents of single tracheal myocytes, an effect reverted by propranolol and ICI 118,551, and mimicked by 0.1microM salbutamol. Thus, we concluded that alpha-methyl-5-HT activates beta(2)-adrenoceptors in guinea pig tracheal smooth muscle at concentrations >or=10microM. This effect must be taken into account when this drug is used in airway smooth muscle and in other tissues expressing beta(2)-adrenoceptors. PMID- 17079162 TI - Anti-inflammatory activity of fisetin in human mast cells (HMC-1). AB - Mast cells play an important role in the pathogenesis of allergic diseases through the release of inflammatory mediators such as histamine, cysteinyl leukotrienes, cytokines, and chemokines. Flavonoids, like fisetin are naturally occurring molecules with antioxidant, cytoprotective, and anti-inflammatory actions. The aim of our study was to examine whether fisetin modulates inflammatory reaction in stimulated human mast cells (HMC-1). Fisetin decreased phorbol-12-myristate 13-acetate plus calcium ionophore A23187 (PMACI)-stimulated gene expression and production of tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, IL-4, IL-6, and IL-8 in HMC-1 cells. Fisetin inhibited PMACI-induced phosphorylation of p38 mitogen-activated protein kinase, extracellular-regulated kinase, and c-Jun N-terminal kinase. In addition, fisetin suppressed nuclear factor (NF)-kappaB activation induced by PMACI, leading to expression of IkappaB-alpha phosphorylation and degradation. Fisetin suppressed powerful induction of NF-kappaB promoter-mediated luciferase activity. These pharmacological actions of fisetin produce new suggestion that fisetin is a potential medicine for treatment of inflammatory diseases through the down regulation of mast cell activation. PMID- 17079163 TI - Attenuated cardioprotective response to bradykinin, but not classical ischaemic preconditioning, in DOCA-salt hypertensive left ventricular hypertrophy. AB - Hypertensive left ventricular hypertrophy (LVH) co-exists frequently with ischaemic heart disease. While ischaemic preconditioning (IPC) is known to protect against ischaemia-reperfusion injury in LVH, it is not known if other cardioprotective manoeuvres are effective. Bradykinin, a key autacoid mediator in IPC, is protective in normal hearts but its ability to protect against ischaemia reperfusion injury in LVH is unknown. Hypertensive LVH was induced in male rats by 4 weeks treatment with deoxycorticosterone acetate (DOCA) and salt drinking fluid. Hearts were Langendorff perfused, subjected to 35 min coronary artery occlusion and 120 min reperfusion, and infarct size (AN/RZ %) was determined by tetrazolium staining. The effects of IPC with 2 x 5 min cycles of global ischaemia or 10 min pretreatment with bradykinin were assessed. DOCA-salt rats were markedly hypertensive and left ventricle/body weight ratio was 26% greater than in normotensive controls. Baseline coronary flow and risk zone/LV ratio were similar in normotensive hearts and DOCA-salt hearts, and infarct size was similar (AN/RZ 50.6+/-3.2% and 47.0+/-3.1%, respectively). IPC was equally protective in normotensive and DOCA-salt hearts (AN/RZ 18.6+/-3.3% and 18.4+/-2.3%, respectively, P < 0.01 versus corresponding control). Bradykinin 0.1, 0.2 or 0.5 microM pretreatment produced concentration-dependent infarct limitation in normotensive hearts (bradykinin 0.5 microM AN/RZ, 9.5+/-3.6%, P < 0.01 versus normotensive control), but the effect in DOCA-salt hearts was attenuated (bradykinin 0.5 microM AN/RZ, 23.4+/-3.8%). Further, the pre-ischaemic coronary vasodilator response to bradykinin was abrogated in DOCA-salt hypertensive hearts. We conclude that the cardioprotective action of bradykinin is markedly attenuated in moderate LVH and coronary vasodilator effect is lost. The reasons for reduced sensitivity to bradykinin in the hypertensive heart are unknown but these findings may have implications for the application of preconditioning mimetic interventions in LVH. PMID- 17079164 TI - Production and characterization of human CTLA4Ig expressed in transgenic rice cell suspension cultures. AB - Human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4I g) fusion protein, a novel immunosuppressive agent, was expressed in transgenic rice cell suspension culture and its characteristics and in vitro activities were investigated. The expression vector pMYN409 was constructed to express hCTLA4I g under the control of rice alpha-amylase 3D (RAmy3D) promoter. Transgenic calli were prepared by particle bombardment mediated transformation and were screened for hCTLA4I g expression using ELISA. Under the induction condition by sugar starvation, suspension-cultured rice cells secreted hCTLA4I g into the media up to 31.4 mg/L in flask culture. The rice-derived hCTLA4Ig (hCTLA4IgP) was purified from the culture media with affinity chromatography using protein A and compared with CHO derived hCTLA4Ig (hCTLA4IgM). Recombinant hCTLA4IgP has molecular weight of approximately 50 kDa on SDS-PAGE under reducing condition, which is a little different from that of hCTLA4IgM probably due to the difference of carbohydrate chain structures. Purified hCTLA4IgP was biologically active and was confirmed to suppress T-cell proliferation. PMID- 17079165 TI - Expression, purification, and in vitro refolding of soluble tumor necrosis factor related apoptosis-inducing ligand (TRAIL). AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a new member of the TNF superfamily. Here, a recombinant form of the extracellular domain of the TRAIL (sTRAIL) was expressed in Escherichia coli BL21(DE3) under the control of a T7 promoter. The resulting insoluble bodies were separated from cellular debris by centrifugation and solubilized with 8 M urea. A rapid and simple on column refolding procedure was developed. It was applied and then the refolded sTRAIL was purified by anion-exchange chromatography. The purified final product was >98% pure by SDS-PAGE stained with Coomassie brilliant blue R-250. Mass spectroscopic analysis indicated the protein to be 19.2 kDa, which equalled the theoretically expected mass. N-terminal sequencing of refolding sTRAIL showed the sequence which corresponded to the designed protein. The renatured protein displayed its immunoreactivity with the antibodies to TRAIL protein by Western blotting. The purified sTRAIL had a strong cytotoxic activity against human cervical cancer HeLa cells with ED50 about 1.5 mg/L. Circular dichroism and fluorescence spectrum analysis showed that the refolded sTRAIL had a structure similar to that of native protein with beta-sheet secondary structure. This efficient procedure of sTRAIL renaturation may be useful for the mass production of this therapeutically important protein. PMID- 17079166 TI - Effect of sex hormones on neuromuscular control patterns during landing. AB - The purpose of the study was to investigate the effects of sex hormones across menstrual cycle phases on lower extremity neuromuscular control patterns during the landing phase of a drop jump. A repeated-measures design was utilized to examine sex hormone effects in 26 recreationally active eumenorrheic women. Varus/valgus knee angle and EMG activity from six lower extremity muscles were recorded during three drop jumps from a 50 cm platform in each phase of the menstrual cycle. Blood assays verified sex hormone levels and cycle phase. The semitendinosus muscle exhibited onset delays (p0.006) relative to ground contact during the luteal phase, and demonstrated a significant (p0.05) difference between early and late follicular phases. Muscle timing differences between the gluteus maximus and semitendinosus were decreased (p0.05) in the luteal compared to early follicular phases. These results suggest a different co-contractive behavior between the gluteus maximus and semitendinosus, signifying a shift in neuromuscular control patterns. It appears that female recreational athletes utilize a different neuromuscular control pattern for performing a drop jump sequence when estrogen levels are high (luteal phase) compared to when they are low (early follicular phase). PMID- 17079167 TI - A neural plasticity perspective on the schizophrenic condition. AB - Imbalanced plasticity of neural networks in the brain is proposed to underlie deficits in the integration of efferent and afferent processes in schizophrenia. These deficits affect the priming of the behavior implementing systems by prior knowledge, and thus impair both controlled regulation and automatic activation of mental and motor processes. The sense of self as a distinct entity can consequently be undermined. In predominantly reality-distorting patients, hypo plasticity of neural connectivity may cause the emergence of highly focused but inflexible patterns of activation in their representation and response systems. This may lead to dominance of prepotent patterns of activity in these systems and a relative inability of higher control systems to bias lower level activity towards congruence with the ongoing cognitive and motor context. By contrast, predominantly disorganized patients are characterized by hyper-plastic connectivity. This leads to a weakening of prepotent response tendencies but also, as in reality-distorting patients, to less effective top-down contextual constraining. PMID- 17079168 TI - Reinstatement of episodic-like memory in rats by neurokinin-1 receptor antagonism. AB - We previously showed that a systemic administration of the selective non-peptide neurokinin-1-receptor (NK-1-R) antagonist SR140333 increases hippocampal acetylcholine levels and facilitates long term memory. In the present study, we investigated whether systemic SR140333 has beneficial effects on episodic-like memory for unique experiences. Rats received either no injection, a vehicle injection or SR140333 at doses of 1, 3 and 9 mg/kg (i.p.) prior to the acquisition of an object memory for what, where and when. In line with previous results, untreated rats showed episodic-like memory, while vehicle-injected rats were impaired. A low dose of 1mg/kg SR140333 reinstated episodic-like memory. This result might be related to the effects of SR140333 on hippocampal cholinergic transmission and/or on the stress-response elicited by the injection procedure. Higher doses of SR140333 (3 and 9 mg/kg) induced psychomotor effects, including stereotypic behaviors and arched posture. Since NK-1-R antagonists have anxiolytic and promestic properties and induce hippocampal acetylcholine release at lower doses, they might be effective in the alleviation of the cognitive deficits and increased anxiety seen in early stages of Alzheimer's disease. PMID- 17079170 TI - "Despair" induced by extinction trials in the water maze: relationship with measures of anxiety in aged and adult rats. AB - We have previously reported that extinction of escape behavior in the water maze due to the removal of the platform coincided with the development of behavioral "despair" in aged and adult rats, as assessed by immobility. The present study examines further predictions derived from the hypothesis that the withholding of reinforcement induces behaviors akin to depression. We tested for correlations between extinction performance and immobility, as well as between immobility and measures of anxiety in aged and adult rats. Age comparisons were also performed on these variables. Forty aged and 29 adult male Wistar rats (24 and 3 months old, respectively) were examined in the open field, black/white box and elevated plus maze followed by 6 days of training in the water maze hidden platform task and 8 days of extinction without the platform. Indices of immobility increased over trials of extinction, with the aged showing higher levels, earlier onsets and larger slope increases of immobility than the adults. A lower resistance-to extinction was predictive of more "despair" in both age groups. Between-group differences in the open field, black/white box and elevated-plus maze indicated that the aged showed more anxiety-like behavior than the adults and/or explored these environments less. Within the aged group, indicators of fearfulness in the three tests were predictive of higher levels of "despair". The extinction-despair model is held to provide the promise of a conceptual and empirical model of human depression that is the consequence of withdrawal of reinforcement. PMID- 17079169 TI - Involvement of the rostral anterior cingulate cortex in consolidation of inhibitory avoidance memory: interaction with the basolateral amygdala. AB - Previous findings suggest that the rostral anterior cingulate cortex (rACC) is involved in memory for emotionally arousing training. There is also extensive evidence that the basolateral amygdala (BLA) modulates the consolidation of emotional arousing training experiences via interactions with other brain regions. The present experiments examined the effects of posttraining intra-rACC infusions of the cholinergic agonist oxotremorine (OXO) on inhibitory avoidance (IA) retention and investigated whether the BLA and rACC interact in enabling OXO effects on memory. In the first experiment, male Sprague-Dawley rats were implanted with bilateral cannulae above the rACC and given immediate posttraining OXO infusions. OXO (0.5 or 3 ng) induced significant enhancement of retention performance on a 48-h test. In the second experiment, unilateral posttraining OXO infusions (0.5, 3.0 or 10 ng) enhanced retention when infused into rACC, but not caudal ACC, consistent with previous evidence that ACC is composed of functionally distinct regions. A third experiment investigated the effects of posttraining intra-rACC OXO infusions (0.5 or 10 ng) in rats with bilateral sham or NMDA-induced lesions of the BLA. The BLA lesions did not impair IA retention, but blocked the enhancement induced by posttraining intra-rACC OXO infusions. Lastly, unilateral NMDA lesions of rACC blocked the enhancement of IA retention induced by posttraining ipsilateral OXO infusions into the BLA. These findings support the hypothesis that the rACC is involved in modulating the storage of emotional events and provide additional evidence that the BLA modulates memory consolidation through interactions with efferent brain regions, including the cortex. PMID- 17079171 TI - Extinction learning, reconsolidation and the internal reinforcement hypothesis. AB - Retrieving a consolidated memory--by exposing an animal to the learned stimulus but not to the associated reinforcement--leads to two opposing processes: one that weakens the old memory as a result of extinction learning, and another that strengthens the old, already-consolidated memory as a result of some less well understood form of learning. This latter process of memory strengthening is often referred to as "reconsolidation", since protein synthesis can inhibit this form of memory formation. Although the behavioral phenomena of the two antagonizing forms of learning are well documented, the mechanisms behind the corresponding processes of memory formation are still quite controversial. Referring to results of extinction/reconsolidation experiments in honeybees, we argue that two opposing learning processes--with their respective consolidation phases and memories--are initiated by retrieval trials: extinction learning and reminder learning, the latter leading to the phenomenon of spontaneous recovery from extinction, a process that can be blocked with protein synthesis inhibition. PMID- 17079172 TI - Anatomy and ultrasonographic appearance of the tympanic bulla and associated structures in the rabbit. AB - Otitis media is a relatively common condition in the rabbit although it can be difficult to evaluate clinically. Diagnostic imaging of the rabbit has traditionally involved radiography and there are few reports regarding the use of ultrasound. The aim of this study was to determine if the tympanic bulla (TB) and associated structures could be evaluated in the rabbit using currently available ultrasound equipment. The normal anatomy of the region was established using gross dissection and emascerated specimens and compared with the dog and cat. Ten New Zealand white rabbit cadavers were examined using two ultrasound machines and four transducers from a variety of locations. A 12 MHz linear transducer was found to be most appropriate. From a lateral approach the external ear canal could be visualized to the level of the external acoustic meatus while the TB itself could only be visualized from a ventral approach. The jugular and mastoid processes in the rabbit are very prominent and interfered with imaging of the TB from all approaches. Water-soluble lubricant introduced into the TB could be identified through the ventral bone wall of the TB and allowed the lumen and far wall to be visualised. Further work is required to determine whether ultrasound examination of the TB can be performed in live rabbits and to evaluate its benefits in the evaluation of clinical cases of otitis media. PMID- 17079173 TI - Prevalence and clinical phenotype of the p.Val226Met glucokinase gene mutation in French Canadians in Quebec, Canada. AB - Our objectives were to describe the clinical phenotype of maturity-onset diabetes of the young (MODY) type 2 in a group of French Canadians and estimate its prevalence in this population. Index cases were identified by an abnormal fasting blood glucose (FBG) upon metabolic evaluation for dyslipidemia. Mutational analyses confirmed that all probands and affected family members were positive for the same glucokinase mutation, p.Val226Met. The prevalence of this mutation was estimated from a representative sample of French Canadians. Eleven individuals in 5 different families were diagnosed with MODY 2. Four of the five families originated from the same region in Quebec. In affected children (n = 6), the median age at diagnosis was 7.6 years (range = 2.9-9.4). All were asymptomatic. The range of FBG was 4.4-7.0 mmol/L; 5 out of the 6 pediatric patients had normal FBG values during the course of follow-up. One child presented with consistently normal FBG. Four of the adults who screened positive for MODY 2 had been previously misdiagnosed with type 2 DM, and one female had a history of gestational DM. The estimated prevalence of heterozygotes for the p.Val226Met mutation in French Canadians was 0.057% (95%CI 0.01-0.32%). In conclusion, this report presents the first confirmed case of MODY 2 with persistently normal FBG. In children and adolescents, a normal FBG does not allow for the exclusion of a MODY 2 diagnosis. Our results are consistent with a founder effect for the p.Val226Met glucokinase gene mutation in Quebec, Canada. PMID- 17079174 TI - Detection of an Alu insertion in the POMT1 gene from three French Walker Warburg syndrome families. AB - Walker Warburg syndrome (WWS) is the most severe of a group of multiple congenital disorders known as lissencephaly type II ( LIS Type II) associated with congenital muscular dystrophy and eye abnormalities. The POMT1 gene is the most frequently affected found in 20% of patients with WWS. We describe five fetuses with WWS in three non-related families carrying a same mutation in the POMT1 gene. All fetuses presented with tetra ventricular hydrocephaly, and arachnoidal neuroglial ectopia and cortical dysplasia characteristic of LIS type II. We performed sequencing of the POMT1 gene on fetal DNA. The five fetuses were found to share an insertion of an inversed Alu repeated DNA element within exon 3 of the POMT1 gene, all at the heterozygous state except one at the homozygous state. This mutation was associated with a common transition c.2203 C > T (p.Arg735Cys) in exon 20 on the same allele and similar intragenic haplotype, suggesting that the three families could be related or indicating a possible founder effect in France. Insertions of Alu sequences, which are rarely found in coding regions, have occasionally been reported to cause other genetic diseases. However, this is the first report of a retrotransposon insertion in the POMT1 gene associated with WWS. PMID- 17079175 TI - Generation of a conditional knockout of murine glucocerebrosidase: utility for the study of Gaucher disease. AB - Gaucher disease is a disorder of sphingolipid metabolism resulting from an inherited deficiency of the lysosomal hydrolase glucocerebrosidase. Affected individuals present with a spectrum of clinical symptoms ranging from hepatosplenomegaly, haematological abnormalities, and bone pain in type 1 disease, to severe neurodegeneration and premature death in types 2 and 3 disease. Although the basic biochemical defect is well characterized, there remains a poor understanding of the underlying pathophysiology of disease. In vitro studies suggest that macrophage glucocerebroside storage leads to tissue dysfunction through complex mechanisms involving altered intracellular calcium homeostasis and apoptosis. In order to study the pathogenic roles of these complex interactions, a viable animal model for Gaucher disease is needed. The complexity of this single gene disorder has been emphasized by the varied results of previous murine Gaucher models, ranging from perinatal lethality to phenotypically and biochemically asymptomatic animals. Recognizing the need to modulate the biochemical phenotype in mice to produce a relevant model, we have created a murine strain with key exons of the glucocerebrosidase gene flanked by loxP sites. We show that expression of Cre-recombinase in cells of hematopoietic and endothelial origin results in deficiency of glucocerebrosidase in the liver, spleen, bone marrow, and peripheral white cells. Glucocerebroside storage in this model leads to progressive splenomegaly with Gaucher cell infiltration and modest storage in the liver by 26 weeks of age. These results indicate the utility of this loxP GBA targeted murine strain for understanding the complex pathophysiology of Gaucher disease. PMID- 17079176 TI - The long-term international safety experience of imiglucerase therapy for Gaucher disease. AB - BACKGROUND: Gaucher disease is a lysosomal storage disorder resulting from a deficiency of the lysosomal enzyme glucocerebrosidase. Since approval by the FDA in 1994 and EMEA in 1997, enzyme replacement therapy with Cerezyme (imiglucerase for injection) has been the standard of care for the treatment of Gaucher disease. OBJECTIVE: To review the long-term international safety experience of imiglucerase from 1994 through 2004. MATERIALS AND METHODS: All spontaneous adverse event reports captured in the pharmacovigilance database for imiglucerase from 1994 through 2004 were analyzed. All adverse events were classified using the current version of the Medical Dictionary for Regulatory Activities (MedDRA). Patients without prior exposure to imiglucerase from 1994 through 2005 were assessed for the development of antibodies to imiglucerase as detected by enzyme linked immunosorbant and radioimmunoprecipitation assays. RESULTS: Analysis of the long-term safety experience with imiglucerase therapy demonstrates a stable and low rate of adverse events and seroconversion from 1994 through 2005. The majority of frequently reported adverse events related to imiglucerase were infusion-associated reactions which were predominantly self-limiting in nature and did not require discontinuation of treatment. Between 1994 and 2005, IgG antibodies to imiglucerase were detected in approximately 15% of treatment-naive patients. CONCLUSIONS: The long-term stability of reported events and seroconversion is a reflection of a well-characterized cell expression system and a mature quality-controlled manufacturing process. Imiglucerase is a safe therapy for the treatment of Gaucher disease with a stable and low rate of reported adverse events and seroconversion. PMID- 17079177 TI - [Comparison of transobturator tape (TOT) and tension-free vaginal tape (TVT) using perineal ultrasound]. PMID- 17079178 TI - [Laparoscopic surgery of deep endometriosis. About 118 cases]. PMID- 17079179 TI - [Mode of delivery of HIV-infected women: a retrospective study of 358 pregnancies followed in the same hospital between 2000 and 2004]. PMID- 17079180 TI - [Repeated spontaneous abortions: from onirism to fantasy]. PMID- 17079181 TI - Aspects of ultrasonically assisted transesterification of various vegetable oils with methanol. AB - The batch transesterification of vegetable oil with methanol, in the presence of potassium hydroxide as catalyst, by means of low frequency ultrasound (40 kHz) was studied with the aim of gaining more knowledge on intimate reaction mechanism. The concentration of fatty acid methyl esters, of mono-, di- and triglycerides of the actual reaction mixture were determined at short reaction time by HPLC. The effect of ultrasounds on the lipids transesterification correlated with triglyceride structures is discussed. It was found that under ultrasonic activation the rate-determining reaction switches from DG-->MG (classical mechanic agitation) to MG+ROH-->Gly+ME (ultrasonically driven transesterification). PMID- 17079182 TI - Regulation of corneal angiogenesis in limbal stem cell deficiency. AB - Corneal angiogenesis is associated with a variety of corneal diseases, and is sometimes vision threatening. In recent years, with the discovery of major pro- and anti-angiogenic factors in the cornea, details of the angiogenic process are gradually unveiled. Of note, corneal inflammation and neovascularization associated with severe limbal stem cell (LSC) deficiency is a clinically challenging issue in that the condition persists long after the initial insult, and will not improve without transplantation of LSCs. However, to date the molecular mechanism by which LSC transplantation restores corneal avascularity is not fully understood. In addition to discussing major pro-angiogenic factors involved in corneal neovascularization, this review article also focuses on possible molecular mechanisms underlying persistent inflammation and neovascularization following severe LSC deficiency, and anti-angiogenic factors expressed by human limbo-corneal epithelial cells (HLCECs). Most of the recently discovered corneal anti-angiogenic factors belong to extracellular matrix proteins that acquire angio-inhibitory activity only after proper proteolytic processing. Our recent findings showed that the secretion of endostatin (derived from basement membrane collagen XVIII) and restin (from collagen XV) by HLCECs were enhanced when HLCECs were cultivated on amniotic membrane (AM). This adds to the advantage of transplanting ex vivo expanded HLCECs cultivated on AM in that the anti-angiogenic activity of the epithelial cells is augmented in a physiological way. Furthermore, proteomic profiling of HLCECs and human conjunctival epithelial cells (HCECs) identified a 14-3-3 protein (stratifin) preferentially expressed by HLCECs. In addition to functioning as a cell cycle controller, keratinocyte-derived stratifin induces MMPs which are involved in the generation of restin (by MMP-1) and endostatin (by MMP-3). These findings highlight the significance of delicate epithelial-matrix interactions in the maintenance of corneal avascularity. PMID- 17079183 TI - A deceptive case of gunshot entry wounds -- beware of frangible bullets. AB - In December 2003, two young men decided to go shooting in the countryside near Montpellier, France. One accidentally shot the other. Upon crime scene examination and autopsy of the victim experts observed, at the right thorax, two round wounds, distanced 5mm from each other, presenting typical characteristics of entry wounds of bullets shot from a distance. Because of the presence of two clear cut round wounds, the Procurator suspected voluntary homicide. However, thanks to the balistics expertise, the authors concluded that all fragments belonged to a unique projectile, 22 Short caliber (Remington cartouche) of the frangible type. The barrel of the rifle infact presented an imperfection where the screw was fixed on the frontsite. The screw had obviously been changed, and the new screw was longer and therefore extended into the barrel, causing a small obstacle to the bullet when exiting the barrel. Shooting tests were performed, allowing the authors to conclude that a single bullet had fragmented before entering the body into two fragments. This lead the Procurator to consider the lesions consistent with an accident. PMID- 17079184 TI - Automatic elastic image registration by interpolation of 3D rotations and translations from discrete rigid-body transformations. AB - We present an algorithm for automatic elastic registration of three-dimensional (3D) medical images. Our algorithm initially recovers the global spatial mismatch between the reference and floating images, followed by hierarchical octree-based subdivision of the reference image and independent registration of the floating image with the individual subvolumes of the reference image at each hierarchical level. Global as well as local registrations use the six-parameter full rigid body transformation model and are based on maximization of normalized mutual information (NMI). To ensure robustness of the subvolume registration with low voxel counts, we calculate NMI using a combination of current and prior mutual histograms. To generate a smooth deformation field, we perform direct interpolation of six-parameter rigid-body subvolume transformations obtained at the last subdivision level. Our interpolation scheme involves scalar interpolation of the 3D translations and quaternion interpolation of the 3D rotational pose. We analyzed the performance of our algorithm through experiments involving registration of synthetically deformed computed tomography (CT) images. Our algorithm is general and can be applied to image pairs of any two modalities of most organs. We have demonstrated successful registration of clinical whole body CT and positron emission tomography (PET) images using this algorithm. The registration accuracy for this application was evaluated, based on validation using expert-identified anatomical landmarks in 15 CT-PET image pairs. The algorithm's performance was comparable to the average accuracy observed for three expert-determined registrations in the same 15 image pairs. PMID- 17079185 TI - Do children really confuse appearance and reality? AB - Our understanding of many mental, social and physical phenomena hinges on a general understanding that appearances can differ from reality. Yet young children sometimes seem unable to understand appearance-reality dissociations. In a standard test, children are shown a deceptive object and asked what it really is and what it looks like. Many preschool children give the same answer to both questions. This error has been attributed to children's inflexible conceptual representations or inflexibility in representing their own changing beliefs. However, evidence fails to support either hypothesis: new tests show that young children generally understand appearance-reality discrepancies as well as fantasy reality distinctions. These tests instead implicate children's failure to understand the unfamiliar discourse format of the standard test. This misunderstanding might reveal a subtler difficulty in making logical inferences about questions. PMID- 17079186 TI - Tracheal incision for elective tracheotomy in oral cavity cancer. AB - The aim of this prospective study was to compare operative procedure and postoperative complications between horizontal tracheal incision and window-type tracheal excision for elective tracheotomy in patients with oral cavity cancer. Between February 2003 and April 2004, 40 patients with advanced-stage oral cavity cancer were consecutively seen at our tumor clinic. All patients underwent elective tracheotomy before wide excision of the tumor and free flap reconstruction. Either horizontal tracheal incision (H group) or window-type tracheal excision (W group) was randomly carried out in two groups comprising 20 patients each. The post-tracheotomy tracheal stenosis was evaluated by tracheal computed tomography (CT) coupled with advantage workstation 4.0 software for reconstruction. Both groups had the following similar characteristics: age, gender, tracheotomy days, or interval between decannulation and CT scan evaluation. However, the incision time was statistically less in the H group (16.4s) compared with the W group (43.4s). Tracheal stenosis was found in five patients (25%) in the H group and four patients (20%) in the W group, with no significant differences. Nevertheless, neither dyspnea or stridor after decannulation, difficulty in tube exchange, nor accidental extrusion was reported in either groups. The horizontal incision for elective tracheotomy is simpler and faster than the window-type tracheal excision. The complication rate is not significantly different in both groups. We therefore, recommend using horizontal incision for elective tracheotomy in patients with oral cavity cancer. PMID- 17079187 TI - Comment on 'Bone SPECT reduces the number of unnecessary mandibular resections in patients with squamous cell carcinoma' [Van Can EM, Oyen WJG, Koole R, Stoelinga PJW. Oral Oncology 2006;42:409-14]. PMID- 17079188 TI - Advances and perspectives in Leishmania cell based drug-screening procedures. AB - Efforts for the development of new therapeutics, essential for the control of leishmaniasis rely mainly on screening of potentially effective compounds in pathogen growth/multiplication assays, both in vitro and in vivo. Screenings designed to closely reflect the situation in vivo are currently labor-intensive and expensive, since they require intracellular amastigotes and animal models. Screenings designed to facilitate rapid testing of a large number of drugs are not performed on the clinically relevant parasite stage, but the promastigotes. The ability to select transgenic Leishmania expressing reporter proteins, such as the green fluorescent protein (GFP) or the luciferase, opened up new possibilities for the development of drug screening tests. In this review we will focus on available methodologies for direct drug screening purposes against the mammalian stage of the parasite, with emphasis on the future developments that could improve sensitivity, reliability, versatility and the throughput of the intracellular model screening. PMID- 17079189 TI - The development of heart failure in patients with stable angina pectoris. AB - BACKGROUND: To describe the clinical characteristics of patients with stable angina pectoris who develop heart failure and the events preceding its onset. METHODS AND RESULTS: Of 7665 patients with stable angina in the ACTION trial, which compared long-acting nifedipine to placebo, 207 (2.7%) developed heart failure (HF) during a mean follow-up of 4.9 years. Those who developed HF were significantly (P<0.05) older, more often had diabetes, had a more extensive history of cardiovascular disease, lower ejection fractions, a higher serum creatinine and glucose, a lower haemoglobin, and were more often on blood pressure lowering drugs. A cardiac event or an intervention (n=155), a significant non-cardiac infection (n=19) or poor control of hypertension (n=12) preceded the development of HF in 186/207 cases (90%). There was no obvious precipitating factor in the remaining 21 patients (10%). Myocardial infarction increased the risk of the development of new HF within one week more than 100 fold. Nifedipine reduced the incidence of HF by 29% (P=0.015). CONCLUSIONS: The development of heart failure is uncommon in patients with stable angina, and even less so in the absence of an obvious precipitating factor. PMID- 17079190 TI - Usefulness of bifocal pacing in patients with heart failure and intraventricular conduction delay. AB - BACKGROUND: Bifocal pacing (BFP) has been proposed as a feasible alternative to cardiac resynchronization therapy. AIM: To evaluate BFP in patients with severe heart failure and significant intraventricular conduction delay and to compare it with biventricular pacing (BVP). METHODS: Echocardiographic examination including TDI and invasive measurement of haemodynamics was performed under basal conditions, during BFP and during BVP. RESULTS: Fifty patients were included: 29 with ischaemic heart disease (IHD), 21 with idiopathic dilated cardiomyopathy (IDCM). LV dp/dt(max) increased during BFP compared to the basal state (13.4%, 95% CI 9.2-17.6%, p<0.0001) and a further increase was achieved during BVP (29.5%, 95% CI 23.7-35.4%, p<0.0001). A significant correlation was found between the distance of the right ventricular apical and outflow tract leads and percentage of dp/dt(max) increase in IDCM patients (r=0.72), but not in IHD patients. Interventricular mechanical delay (IVMD) decreased in BFP (43+/-22 ms vs. 53+/-31 ms, p=0.006), but BVP produced even shorter IVMD (22+/-19 ms, p<0.0001). In all patients, regional systolic contraction times were significantly shortened, corresponding with prolongation of the respective regional diastolic filling times during both BFP (p<0.05 for all segments) and BVP (p<0.001 for all segments). CONCLUSIONS: BFP improves LV haemodynamics by decreasing the inter- and intraventricular conduction delays. The leads in the right ventricle should be placed at the longest achievable distance. BVP is superior to BFP. PMID- 17079191 TI - Psychiatric and behavioral side effects of the newer antiepileptic drugs in adults with epilepsy. AB - OBJECTIVE: Psychiatric/behavioral side effects (PSEs) are common in patients taking antiepileptic drugs (AEDs). The objective of the study described here was to compare the PSE profiles of the newer AEDs. METHODS: We examined the charts of 1394 adult outpatients seen at the Columbia Comprehensive Epilepsy Center who had taken one of the newer AEDs. We compared the rate of AED-related PSEs in patients newly started on the newer AEDs both before and after controlling for non-AED predictors of PSEs. RESULTS: Overall, 221 of 1394 (16%) patients experienced PSEs. The average rate of AED-related PSEs for a single AED was 8.4%, with 6.1% resulting in dosage change and 4.3% resulting in AED discontinuation. Significantly fewer PSEs were attributed to gabapentin (n=160, 0.6% incidence, P<0.001) and lamotrigine (n=547, 4.8% incidence, P<0.001), and significantly more PSEs were attributed to levetiracetam (n=521, 15.7% incidence, P<0.001; 8.8% discontinued LEV because of PSEs). Vigabatrin, felbamate, and oxcarbazepine were associated with similarly low rates of PSEs in many analyses but with fewer of patients. Tiagabine was associated with high PSE rates (similar to those for levetiracetam), but was used much less commonly at our center. Intermediate rates of PSEs were attributed to topiramate and zonisamide (both nonsignificant). Psychiatric history was the most significant nondrug predictor of AED-related PSEs (PSEs occurred in 23% of patients with a psychiatric history vs 12% of patients without such a history, P<0.001). The relative rates of AED-related PSEs were similar when controlling for non-AED predictors and when analyzing only patients on monotherapy. CONCLUSIONS: There are significant differences between the newer AEDs in terms of their PSE profiles. Patients taking levetiracetan experience significantly more PSEs than average, and patients taking gabapentin and lamotrigine experience significantly fewer PSEs. Even with the medication with the highest rate of PSEs (levetiracetam), less than 10% of patients discontinued it because of PSEs. A past psychiatric condition is the most significant nondrug predictor of AED-related PSEs. PMID- 17079192 TI - Evaluation of techniques for increasing recall in a dictionary approach to gene and protein name identification. AB - Gene and protein name identification in text requires a dictionary approach to relate synonyms to the same gene or protein, and to link names to external databases. However, existing dictionaries are incomplete. We investigate two complementary methods for automatic generation of a comprehensive dictionary: combination of information from existing gene and protein databases and rule based generation of spelling variations. Both methods have been reported in literature before, but have hitherto not been combined and evaluated systematically. We combined gene and protein names from several existing databases of four different organisms. The combined dictionaries showed a substantial increase in recall on three different test sets, as compared to any single database. Application of 23 spelling variation rules to the combined dictionaries further increased recall. However, many rules appeared to have no effect and some appear to have a detrimental effect on precision. PMID- 17079193 TI - Attenuation of nitric oxide- and prostaglandin-independent vasodilation of retinal arterioles induced by acetylcholine in streptozotocin-treated rats. AB - Diabetes alters retinal hemodynamics, but little is known about the impact of diabetes on the role of endothelium-derived hyperpolarizing factor (EDHF) in the regulation of retinal circulation. Therefore, we examined how diabetes affects the nitric oxide- and prostaglandin-independent vasodilation of retinal arterioles induced by acetylcholine. Male Wistar rats were treated with streptozotocin (80 mg/kg, i.p.) and experiments were performed 6-8 weeks later. Under artificial ventilation, rats were treated with tetrodotoxin (100 microg/kg, i.v.) to eliminate any nerve activity and prevent movement of the eye. Methoxamine was used to maintain adequate systemic circulation. Fundus images were captured by a digital camera that was equipped with a special objective lens. The vasodilator responses of retinal arterioles were assessed by measuring changes in diameters of the vessels. In streptozotocin-induced diabetic rats and the age-matched controls, acetylcholine increased diameters of retinal arterioles in a dose-dependent manner. The vasodilator responses to acetylcholine in diabetic rats were smaller than those in control rats. The nitric oxide- and prostaglandin-independent vasodilation of retinal arterioles observed under treatment with combination of N(G)-nitro-l-arginine methyl ester (30 mg/kg, i.v.) and indomethacin (5 mg/kg, i.v.) were also attenuated by diabetes. Diabetes did not alter the dilator responses of retinal arterioles to sodium nitroprusside and forskolin. These results suggest that diabetes impairs EDHF-mediated vasodilation of retinal arterioles induced by acetylcholine. The impaired EDHF-mediated vasodilation may contribute to alteration of retinal hemodynamics in diabetes. PMID- 17079194 TI - DNA biosensor based on the electrochemiluminescence of Ru(bpy)3(2+) with DNA binding intercalators. AB - This paper reports a novel detection method for DNA hybridization based on the electrochemiluminescence (ECL) of Ru(bpy)(3)(2+) with a DNA-binding intercalator as a reductant of Ru(bpy)(3)(3+). Some ECL-inducible intercalators have been screened in this study using electrochemical methods combined with a chemiluminescent technique. The double-stranded DNA intercalated by doxorubicin, daunorubicin, or 4',6-diamidino-2-phenylindole (DAPI) shows a good ECL with Ru(bpy)(3)(2+) at +1.19 V (versus Ag/AgCl), while the non-intercalated single stranded DNA does not. In order to stabilize the self-assembled DNA molecules during ECL reaction, we constructed the ECL DNA biosensor separating the ECL working electrode with an immobilized DNA probe. A gold electrode array on a plastic plate was assembled with a thru-hole array where oligonucleotide probes were immobilized in the side wall of thru-hole array. The fabricated ECL DNA biosensor was used to detect several pathogens using ECL technique. A good specificity of single point mutations for hepatitis disease was obtained by using the DAPI-intercalated Ru(bpy)(3)(2+) ECL. PMID- 17079195 TI - DNA/Poly(p-aminobenzensulfonic acid) composite bi-layer modified glassy carbon electrode for determination of dopamine and uric acid under coexistence of ascorbic acid. AB - A nano-composite of DNA/poly(p-aminobenzensulfonic acid) bi-layer modified glassy carbon electrode as a biosensor was fabricated by electro-deposition method. The DNA layer was electrochemically deposited on the top of electropolymerized layer of poly(p-aminobenzensulfonic acid) (Pp-ABSA). Scanning electron microscopy, X ray photoelectron spectroscopy and electrochemical impedance spectrum were used for characterization. It demonstrated that the deposited Pp-ABSA formed a 2-D fractal patterned nano-structure on the electrode surface, and which was further covered by a uniform thin DNA layer. Cyclic voltammetry and electrochemical impedance spectrum were used to characterize the deposition, and demonstrated the conductivity of the Pp-ABSA layer. The biosensor was applied to the detection of dopamine (DA) and uric acid (UA) in the presence of ascorbic acid (AA). In comparison with DNA and Pp-ABSA single layer modified electrodes, the composite bi-layer modification provided superior electrocatalytic actively towards the oxidation of DA, UA and AA, and separated the originally overlapped differential pulse voltammetric signals of UA, DA and AA oxidation at the bare electrode into three well-defined peaks at pH 7 solution. The peak separation between AA and DA, AA and UA was 176 mV and 312 mV, respectively. In the presence of 1.0 mM AA, the anodic peak current was a linear function of the concentration of DA in the range 0.19-13 microM. The detection limit was 88 nM DA (s/n=3). The anodic peak current of UA was also a linear function of concentration in the range 0.4-23 microM with a detection limit of 0.19 microM in the presence of 0.5 mM AA. The superior sensing ability was attributed to the composite nano-structure. An interaction mechanism was proposed. PMID- 17079196 TI - A novel XPC pathogenic variant detected in archival material from a patient diagnosed with Xeroderma Pigmentosum: a case report and review of the genetic variants reported in XPC. AB - The disease Xeroderma Pigmentosum (XP) is genetically heterogeneous and defined by pathogenic variants (formerly termed mutations) in any of eight different genes. Pathogenic variants in the XPC gene are the most commonly observed in US patients. Moreover, pathogenic variants in just four of the genes, XPA, XPC, XPD/ERCC2 and XPV/POLH account for 91% of all XP cases worldwide. In the current study, we describe the clinical, histopathologic, molecular genetic, and pathophysiological features of a 19-year-old female patient clinically diagnosed with XP as an infant. Analysis of archival material reveals a novel variation of a 13 base pair deletion in XPC exon 14 and a previously reported A>C missense pathogenic variant in the proximal splice site for XPC exon 6. Both variations induce frameshifts most likely leading to a truncated XPC protein product. Quantitative RT-PCR also revealed reduced mRNA levels in the archived specimen. Analysis of the XPA, XPD/ERCC2 and XPV/POLH genes in the current specimen failed to reveal pathologic variants. All previously reported pathogenic variants, polymorphisms and known amino acid changes for the XPC gene are compiled and described in the current nomenclature. Given the relative ease of screening for genetic variation and the potential role for such variation in human disease, a proposal for screening appropriate archival materials for alterations in the four most prevalent XP genes is presented. PMID- 17079197 TI - Pegylated interferon and ribavirin in re-treatment of responder-relapser HCV patients. AB - BACKGROUND: The re-treatment of patients who relapse after a course of standard interferon and ribavirin with pegylated interferon alfa-2b plus ribavirin is an open issue. AIMS: To evaluate efficacy and safety of treatment with pegylated interferon alfa-2b plus ribavirin and the role of early HCV-RNA assessment as a predictor of sustained response. PATIENTS: Between May 2001 and December 2002, 242 consecutive patients with chronic hepatitis C were enrolled in an open, regional, multicentre study. Seventy-eight of them were responder-relapsers to a previous course of combination therapy. METHODS: Patients were treated with pegylated interferon alfa-2b (1 microg/kg/week) plus ribavirin (800-1200 mg daily). Qualitative HCV-RNA was performed at week 2. Genotypes 1-4 were treated for 48 weeks, while genotypes 2 and 3 were treated for 24 weeks. RESULTS: We obtained an overall sustained virological response rate of 41.0% (78.6% for patients with genotypes 2-3). CONCLUSION: This treatment schedule prove to be safe and effective in relapsers with genotype non-1 while genotype 1-4 patients had a low rate of sustained virological response. Qualitative virological assessment after 2 weeks may identify patients who are more likely to reach sustained virological response, but it is not a valid tool for a stopping rule approach. PMID- 17079198 TI - Effect of polyethylene pretreatments on the biomimetic deposition and adhesion of calcium phosphate films. AB - The effect of ultraviolet irradiation and glow discharge (GD) processing of the polyethylene (PE) substrates on deposition of calcium phosphate (CaP) films from supersaturated aqueous calcium phosphate solutions was investigated in this study. CaP coatings deposited on the PE substrates were comprised of elongated clusters of spherical particles and 100% of the free surface area of nearly all of the substrates was covered with a porous CaP film after a 3 day immersion. Nano-scratch tests determined that PE-CaP adhesion was most improved when PE substrates were subjected to 50W GD treatments. As determined by contact angle measurements, the GD-treated PE samples had the highest electron donor parameter of surface energy, suggesting that enhancing the electron donor parameter of PE leads to improved adhesion with the biomimetic CaP coating. PMID- 17079200 TI - Platelet-rich plasma preparation using three devices: implications for platelet activation and platelet growth factor release. AB - BACKGROUND: In this study, three commercial systems for the preparation of platelet-rich plasma (PRP) were compared and platelet growth factors release was measured. METHODS: Ten healthy volunteers donated whole blood that was fractionated by a blood cell separator, and a table-top centrifuge to prepare PRP. Furthermore, an autologous growth factor filter was used to concentrate PRP fractionated by the blood cell separator. PRP was subsequently activated with autologously produced thrombin to degranulate the platelets to measure platelet derived growth factor-AB (PDGF-AB), transforming growth factor-beta (TGF-beta), insulin-like growth factor-1 (IGF-1), and vascular endothelial growth factor (VEGF). RESULTS: PRP contained significantly higher platelet counts compared with baseline values (p < 0.001). PDGF-AB concentrations were increased more than 18 fold in the platelet gel supernatant when the cell-separator and GPS were used, whereas only a 3-fold increase was seen with the AGF. CONCLUSION: The three PRP devices enable the preparation of PRP for the release of high concentrations of platelet growth factor, but showed different harvesting capacities for the collection of concentrated platelets. The administration of thrombin for PRP activation resulted in the release of high concentrations of PDGF-AB and TGF-beta but only when PRP had not been activated during the preparation process in vitro. PMID- 17079199 TI - Proximal hip geometry is linked to several chromosomal regions: genome-wide linkage results from the Framingham Osteoporosis Study. AB - INTRODUCTION: Femoral geometry contributes to bone strength and predicts hip fracture risk. The purpose of this study was to evaluate heritability (h(2)) of geometric indices of the proximal hip and to perform whole-genome linkage analyses of these traits, adjusted for body size. METHODS: DXA scans of the proximal femur from 1473 members of 323 pedigrees (age range 31-96 years) from the population-based Framingham Osteoporosis Study were obtained. Using the hip structural analysis program, we measured femoral neck length (FNL, cm) and neck shaft angle (NSA); subperiosteal width (WID, cm), cross-sectional area (CSA, cm(2)); and section modulus (Z, cm(3)) at the narrowest section of the neck (NN), intertrochanteric (IT) and femoral shaft (S) regions. Linkage analyses were performed for the above indices with a set of 636 markers using variance components maximum likelihood method. RESULTS: Substantial genetic influences were found for all geometric phenotypes, with h(2) values between 0.28 (NSA) and 0.70 (IT_WID). Adjustment for height and BMI did not alter h(2) of NSA and FNL but decreased h(2) of the cross-sectional indices. We obtained substantial linkage (multipoint LOD >3.0) for S_Z at 2p21 and 21q11 and S_WID at Xq25-q26. Inclusion of height and BMI as covariates resulted in much lower LOD scores for S_Z, whereas linkage signals for S_Z at 4q25, S_CSA at 4q32 and S_CSA and S_Z at 15q21 increased after the adjustment. Linkage of FNL at 1q and 13q, NSA at 2q and NN_WID at 16q did not change after the adjustment. CONCLUSION: Suggestive linkages of bone geometric indices were found at 1q, 2p, 4q, 13q, 15q and Xq. The identification of significant linkage regions after adjustment for BMI and height may point to QTLs influencing femoral bone geometry independent of body size. PMID- 17079201 TI - Early injection of OP-1 during distraction osteogenesis accelerates new bone formation in rabbits. AB - Distraction osteogenesis (DO) is a surgical technique for generating new bone by applying controlled distraction of two bony segments post osteotomy. A limitation of the technique is the long time required for the new bone to consolidate. We investigated the effect of injecting osteogenic protein 1 (OP-1) at the beginning of distraction in a rabbit model of DO. Regenerate bone was evaluated using radiology, densitometry, micro-computed tomography (microCT) and histomorphometry. Immunohistochemsitry was used to evaluate changes in expression of various ligands, growth factors and receptors following OP-1 treatment. Compared to the control, a two-fold increase in bone volume was apparent for treated groups at 3 weeks post injection. An upregulation of almost all of the 41 genes examined was observed. Results suggested that applying OP-1 early during distraction can accelerate bone formation by the activation of numerous pathways. This study provides further insights on strategies to improve bone regeneration rate in DO. PMID- 17079202 TI - The gene expression profile of PDGF-treated neural stem cells corresponds to partially differentiated neurons and glia. AB - We have previously shown that platelet-derived growth factor AA (PDGF-AA) stimulates the expansion of neuronal progenitors from neural stem cells, but is unable to replace fibroblast-growth factor 2 (FGF-2) as a stem cell mitogen. In the present study, we compared gene expression in neural stem cells that were grown in the presence of FGF-2 and in cells cultured with PDGF-AA or in the absence of growth factor, which induces differentiation. The genetic program elicited by PDGF-AA (156 significantly regulated genes) was not unique, but an intermediate between the ones of FGF-2-cultured stem cells and differentiated cells. These observations are compatible with the hypothesis that PDGF-AA induces a partial differentiation of neural stem cells, which retain the ability to proliferate, rather than acting solely as an instructing agent for neuronal differentiation. Finally, the transcriptional signature of stem cells grown with FGF-2 included a large number of genes over-expressed in gliomas and a core set of conserved genes periodically expressed during the eukaryote cell cycle. PMID- 17079203 TI - The MET receptor tyrosine kinase contributes to invasive tumour growth in rhabdomyosarcomas. AB - The receptor tyrosine kinase MET and its ligand hepatocyte growth factor (HGF), have been implicated in the genesis of the paediatric tumour rhabdomyosarcoma (RMS). Addition of exogenous HGF to RH30 RMS cells enhanced non-chemotactic migration. Stable transfection of dominant negative MET into RH30 cells attenuated Matrigel invasion and in vivo tumour growth. To assess the role of a putative HGF-MET pathway in human RMS, we measured their expression in a panel of 68 human primary tumours. All tumours expressed MET but with a three orders of magnitude variation of expression and 62% of tumours co-expressed HGF. In contrast with other tumour types, neither high-MET expression nor HGF/MET coexpression correlated with metastatic disease. In a microarray screen, we identified CCN1 as being 7.8-fold up regulated following addition of HGF to RH30 cells and in RMS tumours, CCN1 expression correlated with HGF expression. Surprisingly, we identified MET as a consistent feature of embryonal and not alveolar RMS. PMID- 17079204 TI - Intravenous administration of vascular endothelial growth factor improves cardiac performance and inhibits cardiomyocyte apoptosis. AB - This study investigated the effects of vascular endothelial growth factor (VEGF) intravenous administration on cardiac performance and cardiomyocyte apoptosis in a rat model of acute myocardial infarction. Left coronary artery ligation produced extensive myocardial infarction in 48 rats and sham operated in 24 animals. Twenty-four hours after surgery, the rats were randomized to receive VEGF165-heparin (treated group) or heparin-saline (control group) treatment. The sham-operated animals were also to receive VEGF165-heparin (sham group) treatment. VEGF165 (2 microg/ml) with heparin (50 U) or heparin-saline (50 U/ml) was administered daily via the tail vein for 7 and 14 days. Fifty-eight rats survived and included in the study. There were not significant effects of VEGF on hemodynamic parameters in sham animals. As compared with control animals at 9 days after ligation (with 10 rats for each group), rats treated with VEGF had significantly higher maximum rate of left ventricular pressure rise (+ dP/dtmax) or fall ( - dP/dtmax) and microvessel counts, and significantly lower left ventricular end-diastolic pressure (LVEDP) and infarct size. At 16 days after surgery (12, 7 and 9 rats in sham, control and treated groups; respectively), VEGF treatment significantly increased mean arterial pressure (MAP), left ventricular systolic pressure (LVSP), +/- dP/dtmax and microvessel counts, and significantly decreased LVEDP and infarct size. VEGF treatment significantly inhibited cardiomyocyte apoptosis and the expression of p53, Fas and Bax protein, and increased the expression of Bcl-2 protein in myocardium at 9 days after myocardial infarction. PMID- 17079205 TI - Insulin-like growth factor-I stimulates erythropoiesis when administered enterally. AB - BACKGROUND: Insulin-like growth factors I and II (IGF-I and IGF-II) are potent growth factors involved in development. IGF-I stimulates proliferation of erythropoietic progenitors and parenteral IGF-I administration stimulates in vivo erythropoiesis in animals. IGF-I and IGF-II are both present in mammalian milks and when milk-borne, are resistant to neonatal gastrointestinal degradation. Whether milk-borne IGF-I or IGF-II regulates neonatal erythropoiesis in not known. We hypothesized that physiological doses of enteral IGFs stimulate erythropoiesis in suckling rats. METHODS: Eight day-old Sprague Dawley rats were artificially fed for 4 days with rat milk substitute (RMS) or RMS supplemented with physiological levels of IGF-I or IGF-II. Rats fed IGF-I and IGF-II were compared to control RMS. Blood and marrow were collected; measures of red cell mass, measures of erythropoietic stimulus, and indices of iron status were measured. RESULTS: Rats fed IGF-I had higher hemoglobin (Hb) levels (100 +/- 10 g/l), compared to those fed RMS (94 +/- 9) or IGF-II (91 +/- 6), p < 0.001. After IGF-I supplementation, red blood cell counts (RBC) (p < 0.04) and hematocrits (p < 0.002) were also higher. Plasma erythropoietin (Epo) levels, reticulocytes, plasma iron and erythrocyte iron incorporation were similar. CONCLUSION: Intact enteral IGF-I reaches distal erythropoietic tissue resulting in greater red cell mass, but not by increasing plasma Epo levels or by altering cellular iron transport. PMID- 17079207 TI - How not to criticize the precautionary principle. AB - The precautionary principle has its origins in debates about environmental policy, but is increasingly invoked in bioethical contexts. John Harris and Soren Holm argue that the principle should be rejected as incoherent, irrational, and representing a fundamental threat to scientific advance and technological progress. This article argues that while there are problems with standard formulations of the principle, Harris and Holm's rejection of all its forms is mistaken. In particular, they focus on strong versions of the principle and fail to recognize that weaker forms, which may escape their criticisms, are both possible and advocated in the literature. PMID- 17079208 TI - Who's in the business of saving lives? AB - There are individuals, including children, dying needlessly in poverty-stricken third world countries. Many of these deaths could be prevented if pharmaceutical companies provided the drugs needed to save their lives. Some believe that because pharmaceutical companies have the power to save lives, and because they can do so with little effort, they have a special obligation. I argue that there is no distinction, with respect to obligations and responsibilities, between pharmaceutical companies and other types of companies. As a result, to hold pharmaceutical companies especially responsible for saving lives in third world countries is unjustified. PMID- 17079209 TI - Balancing in ethical deliberation: superior to specification and casuistry. AB - Approaches to clinical ethics dilemmas that rely on basic principles or rules are difficult to apply because of vagueness and conflict among basic values. In response, casuistry rejects the use of basic values, and specification produces a large set of specified rules that are presumably easily applicable. Balancing is a method employed to weigh the relative importance of different and conflicting values in application. We argue against casuistry and specification, claiming that balancing is superior partly because it most clearly exhibits the reasoning behind moral decision-making. Hence, balancing may be most effective in teaching bioethics to medical professionals. PMID- 17079210 TI - The concept of underinsurance: a general typology. AB - In a 2002 speech, Mark McClellan, a member of the Council of Economic Advisors at the White House, said that "[I]n the president's vision, all Americans should have access to high-quality and affordable healthcare." However, many healthcare researchers believe that a growing number of Americans are underinsured. Because any characterization of underinsurance will refer to the value judgments of people about what counts as "adequate" and "inadequate" healthcare, the goal of characterizing and measuring the underinsured is difficult to achieve. In this article, I examine the various dimensions of underinsurance, and propose a typology incorporating those dimensions. PMID- 17079211 TI - Calculating risk/benefit in X-linked severe combined immune deficiency disorder (X-SCID) gene therapy trials: the task of ethical evaluation. AB - In response to adverse events in retroviral gene therapy clinical trials conducted in France to correct for X-linked severe combined immune deficiency disorder (X-SCID), an advisory committee of the Food and Drug Administration convened in October 2002, February 2003, and March 2005, to deliberate and provide recommendations for similarly sponsored research in the United States. A similar National Institutes of Health committee met in February 2003. In this article, I review the transcripts and/or minutes of these meetings to evaluate the extent to which the ethical dimension of the research was engaged even as the molecular and clinical evidence was reviewed. I then provide representative ethical arguments to demonstrate the sort of ethical reasoning that should be included as part of the agenda of such committee meetings. PMID- 17079212 TI - Multilayer conformal applicator for microwave heating and brachytherapy treatment of superficial tissue disease. AB - PURPOSE: The purpose of this study was to construct and perform preliminary functionality evaluations of a multilayer conformal applicator with provisions for thermal monitoring, tight conformity and simultaneous microwave heating and brachytherapy treatment of large-area contoured surfaces. MATERIALS AND METHODS: The multilayer conformal applicator consists of thermal monitoring catheters for fibre-optic monitoring of skin temperatures, a waterbolus, a PCB microwave antenna array, a dielectric spacer for brachytherapy considerations, brachytherapy catheters for delivering HDR radiation and an inflatable air bladder for improving conformity to contoured surfaces. The applicator also includes an elastic attachment structure to hold the applicator securely in place on the patient. The conformity of the applicator to irregular surfaces was evaluated through CT imaging of the applicator fitted onto a life-sized human torso phantom. The fluid flow dynamics of the waterbolus, which impact the effectiveness of temperature control, were evaluated with thermometry during a 19 degrees C step change temperature of the circulating water. RESULTS: CT imaging showed improved conformity to the torso phantom surface following the application of gentle inward pressure from inflating the outer air bladder. Only a small number of 1-5 mm sized air gaps separated the conformal applicator and tissue surface. Thermometry testing of the bolus fluid flow dynamics demonstrated temperature uniformity within +/-0.82 degrees C across a 19 x 34 x 0.6 cm area bolus and +/-0.85 degrees C across a large 42 x 32 x 0.6 cm area bolus. CONCLUSION: CT scans of the applicator confirmed that the applicator conforms well to complex body contours and should maintain good conformity and positional stability even when worn on a mobile patient. Thermometry testing of two different waterbolus geometries demonstrated that uniform circulation and temperature control can be maintained throughout large, complex bolus shapes. PMID- 17079213 TI - Reliability of temperature and SAR measurements at oesophageal tumour locations. AB - INTRODUCTION: For treatment of oesophageal cancer, neo-adjuvant locoregional hyperthermia (HT) has been applied in combination with chemotherapy (ChT) +/- radiotherapy (RT) at the institute. Until now, 26 patients were treated within a completed phase I study combining HT with ChT and 29 patients within an ongoing phase II study combining HT with ChT + RT. METHODS: HT was given with the 70 MHz AMC-4 waveguide system. Initially, oesophageal temperatures were measured using multi-sensor thermocouple probes (TCs) inside a nasogastric tube (NT), but the question arose whether these measurements were reliable enough to quantify the achieved tumour temperatures accurately. Presently, TCs are mounted on the outside of an inflatable balloon catheter (BC) for better intra-luminal fixation and better contact with the tumour. During 14 treatment sessions in four patients TCs inside a NT and mounted on a BC were used simultaneously. Data from these 14 treatment sessions were used to compare temperature and Specific Absorption Rate (SAR) measurements ('DeltaT-measurements') using NTs or BCs. To determine the predictive value of the local SAR for the tumour temperatures achieved during treatment, the relation between the initial DeltaT and steady state temperature (SST) was evaluated. RESULTS: There was a strong correlation between the temperature measured in the NT (Ttube) and the temperature measured with a BC (Tballoon): R = 0.88 +/- 0.13. However, Ttube was on average approximately 1 degrees C higher than Tballoon and there was a large variation between the different treatments in the relation between both measurements, rendering Ttube a probably unreliable measure for tumour temperatures. The correlation between the DeltaT measured in the NT (DeltaTtube) and with a BC (DeltaTballoon) was rather weak: R = 0.46 +/- 0.25. The correlation between the initial DeltaT and the SST was much stronger for the BC measurements, R = 0.78 +/- 0.19, than for the NT measurements, R = 0.61 +/- 0.23. Thus, DeltaTballoon has a higher predictive value for the achieved tumour temperatures than DeltaTtube. Both DeltaT and SST were generally higher for the NT measurements than for the BC measurements, suggesting an over-estimation of tumour temperatures. Averaged over all treatments in the phase I trial using a NT (20 treatments) or a BC (45 treatments), T90 was significantly higher when measured with a NT. CONCLUSION: Oesophageal temperature and SAR (DeltaT) measurements inside a NT are less reliable than BC measurements. These artefacts are due to bad thermal contact with the tumour tissue and are, therefore, not specific for thermocouple thermometry. For reliable temperature or SAR measurements inside lumina or cavities good thermal contact must be assured, e.g. by using a balloon catheter. PMID- 17079214 TI - Hepatic radiofrequency ablation at low frequencies preferentially heats tumour tissue. AB - PURPOSE: Radiofrequency ablation is a clinically accepted treatment modality for liver cancer. There are significant differences in dielectric properties between normal and cancer tissue in the liver, which are particularly pronounced at frequencies below 100 kHz. This study performed computer simulations to determine whether radiofrequency (RF) ablation at lower frequencies than currently employed (450-500 kHz) can take advantage of this difference to preferentially deposit energy within the tumour. MATERIALS AND METHODS: Finite Element Method computer models were created for a cooled needle electrode and a multi-tine RF electrode inserted into a 2 cm diameter tumour. RF ablation was simulated and current density as well as tissue temperature distribution determined. In vivo data were used on electrical conductivity of normal and cancer tissue in the models to simulate RF ablation in liver at the currently used frequency of 500 kHz and at 10 kHz. RESULTS: At 500 kHz there was little difference in RF current density and final tissue temperature between normal and cancer tissue. Due to the more pronounced differences in electrical conductivity at 10 kHz, cancer tissue was heated preferentially at this frequency. Depending on power control algorithm, this resulted in either higher intra-tumour temperatures or lower temperatures outside the tumour at 10 kHz compared to 500 kHz. CONCLUSION: Radiofrequency ablation at lower frequencies than currently used may preferentially heat the tumour and preserve normal tissue. A targeted device for selective tumour destruction may be designed to make use of this principle. PMID- 17079215 TI - Heat induced release of Hsp70 from prostate carcinoma cells involves both active secretion and passive release from necrotic cells. AB - PURPOSE: Heat shock protein 70 (HSP70) is released from tumour cells and stimulates a potent anti-tumour immune response. METHODS: This study examined the role of hyperthermia, including heating conditions from the fever range, the hyperthermia range and the thermal ablation range, in HSP70 release from prostate carcinoma cells. It has observed HSP70 release from human prostate carcinoma cell lines (PC-3 and LNCaP) treated with hyperthermia. RESULTS: The effects of hyperthermia were complex and appeared to involve at least two mechanisms for HSP70 release. Hyperthermia at 40 degrees C strongly stimulated HSP70 release by an active secretion pathway. However, as temperatures were increased, this rapid secretion pathway became progressively inhibited and by a temperature of 55 degrees C, active secretion was abolished. However, when cells exposed to these heating conditions were allowed to recover at 37 degrees C for 24 h after heating, HSP70 release was observed at the high ablation temperature range and this appeared to be related to a concomitant damage to the plasma membrane. CONCLUSIONS: Thus, at least two mechanisms contribute to HSP70 release during hyperthermia and the relative contribution from each pathway depends on the temperature conditions. PMID- 17079216 TI - Anti-angiogenic effects of interleukin-12 delivered by a novel hyperthermia induced gene construct. AB - PURPOSE: Interleukin-12 (IL-12) is a pro-inflammatory cytokine possessing anti cancer and anti-angiogenic properties. This study quantitatively assessed the anti-angiogenic effect of IL-12 delivered using an adenoviral vector with murine IL-12 placed under control of a heat shock promoter. This approach limits systemic toxicity by restricting IL-12 delivery locally to the tumour. The kinetics of the downstream cytokines interferon-gamma (IFN-gamma) and interferon inducible protein-10 (IP-10) and other molecules affecting angiogenesis, vascular endothelial growth factor (VEGF) and plasminogen activator inhibitor-1 (PAI-1) were also studied. MATERIALS AND METHODS: 4T1 tumours were grown in Balb/C mice and the AdhspmIL-12 construct was injected intra-tumourally. The tumours were heated after 24 h using a water bath. At various time points post-heating the tumours were collected and quantitatively assessed for cytokine production and vascularity. RESULTS: A significant reduction was seen in the tumour vasculature of the treated group vs. the control group mice. Systemic effects of IL-12 were limited to generalized immunostimulation. No hepatoxicity was noted. CONCLUSIONS: This study suggests that IL-12 can be effectively delivered using a gene-based approach with a heat shock promoter. This results in quantitatively measurable anti-angiogenesis and general immunostimulation. The complex inter-play of other pro- and anti-angiogenic factors (IFN-gamma, IP-10, VEGF and PAI-1) was also studied. PMID- 17079217 TI - A new local ablation for solid malignant tumours: anti-tumour effect of hyperthermal and electrochemical therapy on transplantable mouse cancer. AB - PURPOSE: Hyperthermia induced by electrothermal needle (ETN) and electrochemical therapy (ECT) were combined (HECT) and the anti-tumor effect was evaluated. METHODS: Mice with Sarcoma180 were randomized into four different treatment groups: Control, ECT alone, Hyperthermia alone and HECT. RESULTS: The tumours in the HECT group were completely destroyed and they did not recur within a period of 10 days after treatment. However, tumour recurrence was found in six mice in the Hyperthermia group and five mice in the ECT group. CONCLUSION: The HECT is a potentially effective way to treat solid malignant tumours. PMID- 17079218 TI - Hyperthermia combined with intra-thoracic chemotherapy and radiotherapy for malignant pleural mesothelioma. AB - BACKGROUND: Prognosis for patients with malignant pleural mesothelioma (MPM) remains poor and such patients require intensive treatment. Few studies have examined hyperthermia for MPM. The present study investigated the feasibility of hyperthermia combined with weekly chemo-radiotherapy for patients with MPM and estimated the efficacy of this regimen. METHODS: A total of 11 patients (median patient age was 67 and all had pleural effusion) with MPM were enrolled in this study. The treatment regimen comprised of weekly thermo-radiotherapy with intra thoracic chemotherapy 2-5 times at initiation of treatment. Hyperthermia was performed once per week for approximately 60 min. Hemithorax external radiotherapy was administered once weekly on the same day as hyperthermia and just before thermochemotherapy. Median total radiation dose was 6 Gy (range, 2-10 Gy). Chemotherapy was administered into the thoracic cavity through a tube. Chemotherapeutic agents administered were CDDP for seven patients, carboplatinum (CBDCA) for three patients and both CDDP and CBDCA for one patient. Dose of CDDP was 50 mg/body and dose of CBDCA was 200-300 mg m-2. Response rate and median survival time (MST) and palliative effect were investigated. RESULTS: Complete response was not achieved in any of the 11 patients. Partial response was achieved in three of 11 patients (27.3%), SD in six patients (54.5%) and PD in two patients (18.2%). There was no correlational relationship between thermal parameters and response. MST was 27.1 months. Pleural fluid decreased in all patients after therapy, while all patients displayed improved performance status and could be discharged from hospital. Patients with partial response had a relatively longer survival time than SD or PD. All patients underwent the complete course of treatment and only one of 11 patients developed grade 4 thrombocytopenia. CONCLUSION: It was therefore concluded that hyperthermia combined with intra-thoracic chemotherapy using cisplatinum or carboplatinum may be tolerable. This approach appears effective and more acceptable for patients with MPM with pleural effusion than other multi-modality therapy. PMID- 17079220 TI - Speech and language therapy service delivery for bilingual children: A survey of three cities in Great Britain. AB - BACKGROUND: Speech and language therapy (SLT) managers are expected to ensure that there are appropriate services available for bilingual and multilingual clients in order to ensure an equitable service to all clients. However, there is a paucity of data available to inform service planning. AIMS: To identify the level to which SLT services in three UK cities meet the recommendations of The Royal College of Speech and Language Therapists (RCSLT) Good Practice Guidelines. Socio-demographic information is provided about the number of children from ethnic minorities in the population and the proportion of (bilingual) children from ethnic minorities on the speech and language therapy caseload. Based on this information, it is estimated whether there is proportionate representation of bilingual children on SLT caseloads, and whether services are in place to meet the needs of those clients. METHODS & PROCEDURES: Population statistics were gathered from Census data and data were gleaned from Local Education Authorities. The study used a combination of interview and postal questionnaires to SLTs, with particular emphasis on the issues that may affect service provision. OUTCOMES & RESULTS: As in previous studies, it was difficult to find reliable data. However, two of the three cities studied appeared to be offering a proportionate service to both monolingual and bilingual children in terms of the relative numbers of children on caseloads. Only one city was confident that their SLT service was fully meeting the RCSLT Good Practice Guidelines on bilingualism, although all three cities were aware of them and appeared to be making an effort at varying levels to address the principles of those guidelines. CONCLUSIONS: There is a need for a change in how data on linguistic diversity in society are collected and disseminated, both at a national level and within SLT services, so that informed decisions can influence the future of quality services to minority groups. PMID- 17079221 TI - Effect of sentence length and complexity on working memory performance in Hungarian children with specific language impairment (SLI): A cross-linguistic comparison. AB - BACKGROUND: English-speaking children with specific language impairment (SLI) perform more poorly than their typically developing peers in verbal working memory tasks where processing and storage are simultaneously required. Hungarian is a language with a relatively free word order and a rich agglutinative morphology. AIMS: To examine the effect of linguistic structure on working memory performance. It was examined whether syntactic complexity has a larger impact on working memory performance than sentence length in Hungarian-speaking children, similar to the findings in English speaking children. METHODS & PROCEDURES: In Experiment 1, performance accuracy was measured with two linguistic span tasks that included stimuli with varying sentence length and syntactic complexity. Experiment 2 examined the impact of sentence length and morphological complexity on working memory performance. OUTCOMES & RESULTS: Children with SLI performed more poorly than their age-matched peers in all working memory tasks. Their error patterns differed from those of children with typical language development. Children with SLI produced a high number of interference errors that indicate poor executive functions. The findings were compared with previous results of English-speaking children. Complexity affected working memory performance accuracy differently across languages. In English, it was the increase of syntactic complexity that resulted in a decrease in performance accuracy, whereas in Hungarian, it was the morphological complexity that had a large impact on working memory performance. CONCLUSIONS: Working memory performance depends on the linguistic characteristics of the language tested. In both English- and Hungarian-speaking children, complexity has a larger effect on verbal working memory performance than the length of the stimuli. However, complexity affects working memory performance accuracy differently across languages. PMID- 17079222 TI - Short-term and working memory in specific language impairment. AB - BACKGROUND: Investigations of the cognitive processes underlying specific language impairment (SLI) have implicated deficits in the storage and processing of phonological information, but to date these abilities have not been studied in the same group of children with SLI. AIMS: To examine the extent to which deficits in immediate verbal short-term and working memory may co-occur in a group of children with SLI. METHODS & PROCEDURES: Twenty children aged 7-11 years with SLI completed a comprehensive battery of short-term and working memory, as well as two phonological awareness tasks. OUTCOMES & RESULTS: The majority of the group had deficits in both verbal short-term and working memory, which persisted after the general language abilities of the children were taken into account. A substantial minority showed deficits on visuospatial short-term memory, while impairments of phonological awareness were less marked. CONCLUSIONS: The data indicate dual deficits in verbal short-term and working memory that exceed criterial language abilities characteristic of SLI and may plausibly underpin some of the language learning difficulties experienced by these children. PMID- 17079223 TI - Clinical markers for specific language impairment in Italian: The contribution of clitics and non-word repetition. AB - BACKGROUND: The discovery of clinical markers for specific language impairment (SLI) in children can assist in the accurate identification of children with this disorder, and in a description of the disorder's phenotype for genetic study. One challenge to this type of research is the fact that languages vary in the most salient symptoms of SLI. This study focuses on Italian. AIMS: To determine whether three measures--the use of third-person plural inflections, the use of direct-object clitics and non-word repetition--are successful in distinguishing Italian-speaking children with SLI from their typically developing peers. METHODS & PROCEDURES: Eleven preschool-aged children with SLI, 11 same-age typically developing peers and 11 younger typically developing children participated in the study. The third-person plural inflection and direct-object clitic tasks required the children to describe drawings in response to prompts provided by the examiner. In the non-word repetition task, the children repeated non-words ranging from one to four syllables in length. OUTCOMES & RESULTS: All three measures proved successful either singly or in combination, with direct-object clitics and non-word repetition showing the highest sensitivity and specificity. CONCLUSIONS: Additional research should be pursued to replicate and extend these findings. Along with the potential clinical value of the findings, the results suggest that difficulties with non-final weak syllables--a problem that would adversely affect all three measures--may be an important part of the SLI profile in Italian. PMID- 17079224 TI - Phonological awareness intervention for children with childhood apraxia of speech. AB - AIMS: To investigate the effectiveness of an integrated phonological awareness intervention to improve the speech production, phonological awareness and printed word decoding skills for three children with childhood apraxia of speech (CAS) aged 7;3, 6;3 and 6;10. The three children presented with severely delayed phonological awareness skills before intervention. METHODS & PROCEDURES: In consideration for the heterogeneity in the population with CAS, the study employed a multiple single-subject design with repeated measures. Baseline and post-intervention measures for speech, phonological awareness and decoding were compared. Each child received intervention for three 45-min sessions per week for 3 weeks (approximately 7 h of individual treatment). Sessions focused on developing phoneme awareness, linking graphemes to phonemes and providing opportunities for targeted speech production practice. Phonological awareness activities were linked with each child's speech production goals. OUTCOMES & RESULTS: Two participants significantly improved target speech and phonological awareness skills during intervention. These participants also generalized the phonological awareness skills from trained to untrained items and were able to transfer newly acquired knowledge to improved performance on a non-word reading task. CONCLUSIONS: The results suggest that integrated phonological awareness intervention may be an effective method simultaneously to treat speech production, phonological awareness and decoding skills in some children with CAS. The findings are discussed within the context of the phonological representational theory of CAS. PMID- 17079225 TI - How conceptual frameworks influence clinical practice: Evidence from the writings of John Thelwall, a 19th-century speech therapist. AB - BACKGROUND: The impact of speech therapists' conceptual frameworks on their clinical methods tends to be ignored or taken for granted by today's practitioners. One way to show the importance of such frameworks is to study how they were used previously. John Thelwall, a 19th-century elocutionist, offers a rich source for studying the influence of conceptual frameworks in the past and provides needed distance for clinicians to discover how they frame their own practices. AIMS: To discover how conceptual frameworks have influenced past and current speech-therapy practices. METHODS & PROCEDURES: Content analyses of the writings of Thelwall were performed to discover his conceptual frameworks and how and when they were used. The results were then compared with frameworks in use today. OUTCOMES & RESULTS: Four frameworks were found in Thelwall's writings: medical, linguistic, educational and political. All are still used, but each differs from Thelwall's in scope and detail. These comparisons have the potential for revealing the significance of conceptual frameworks in today's clinical practices. CONCLUSIONS: The ways Thelwall wrote about his work early in the 19th century bear striking resemblances to today's construals of speech therapy practices. It is the scope and application of the frameworks that have changed with time. Both the similarities and differences between current practices and those of Thelwall offer instructive insights into about how frameworks can influence clinical choices and practices. PMID- 17079227 TI - Estrogen-related receptors stimulate pyruvate dehydrogenase kinase isoform 4 gene expression. AB - The pyruvate dehydrogenase complex (PDC) catalyzes the conversion of pyruvate to acetyl-CoA in mitochondria and is a key regulatory enzyme in the oxidation of glucose to acetyl-CoA. Phosphorylation of PDC by the pyruvate dehydrogenase kinases (PDK2 and PDK4) inhibits PDC activity. Expression of the PDK genes is elevated in diabetes, leading to the decreased oxidation of pyruvate to acetyl CoA. In these studies we have investigated the transcriptional regulation of the PDK4 gene by the estrogen-related receptors (ERRalpha and ERRgamma). The ERRs are orphan nuclear receptors whose physiological roles include the induction of fatty acid oxidation in heart and muscle. Previously, we found that the peroxisome proliferator-activated receptor gamma coactivator (PGC-1alpha) stimulates the expression of PDK4. Here we report that ERRalpha and ERRgamma stimulate the PDK4 gene in hepatoma cells, suggesting a novel role for ERRs in controlling pyruvate metabolism. In addition, both ERR isoforms recruit PGC-1alpha to the PDK4 promoter. Insulin, which decreases the expression of the PDK4 gene, inhibits the induction of PDK4 by ERRalpha and ERRgamma. The forkhead transcription factor (FoxO1) binds the PDK4 gene and contributes to the induction of PDK4 by ERRs and PGC-1alpha. Insulin suppresses PDK4 expression in part through the dissociation of FoxO1 and PGC-1alpha from the PDK4 promoter. Our data demonstrate a key role for the ERRs in the induction of hepatic PDK4 gene expression. PMID- 17079228 TI - Protein phosphatase 6 down-regulates TAK1 kinase activation in the IL-1 signaling pathway. AB - TAK1 (transforming growth factor beta-activated kinase 1) is a serine/threonine kinase that is a mitogen-activated protein kinase kinase kinase and an essential intracellular signaling component in inflammatory signaling pathways. Upon stimulation of cells with inflammatory cytokines, TAK1 binds proteins that stimulate autophosphorylation within its activation loop and is thereby catalytically activated. This activation is transient; it peaks within a couple of minutes and is subsequently down-regulated rapidly to basal levels. The mechanism of down-regulation of TAK1 has not yet been elucidated. In this study, we found that toxin inhibition of type 2A protein phosphatases greatly enhances interleukin 1 (IL-1)-dependent phosphorylation of Thr-187 in the TAK1 activation loop as well as the catalytic activity of TAK1. From proteomic analysis of TAK1 binding proteins, we identified protein phosphatase 6 (PP6), a type-2A phosphatase, and demonstrated that PP6 associated with and inactivated TAK1 by dephosphorylation of Thr-187. Ectopic and endogenous PP6 co-precipitated with TAK1, and expression of PP6 reduced IL-1 activation of TAK1 but did not affect osmotic activation of MLK3, another MAPKKK. Reduction of PP6 expression by small interfering RNA enhances IL-1-induced phosphorylation of Thr-187 in TAK1. Enhancement occurred without change in levels of PP2A showing specificity for PP6. Our results demonstrate that PP6 specifically down-regulates TAK1 through dephosphorylation of Thr-187 in the activation loop, which is likely important for suppressing inflammatory responses via TAK1 signaling pathways. PMID- 17079229 TI - Lipid-induced extension of apolipoprotein E helix 4 correlates with low density lipoprotein receptor binding ability. AB - Apolipoprotein E (apoE) serves as a ligand for the low density lipoprotein receptor (LDLR) only when bound to lipid. The N-terminal domain of lipid-free apoE exists as globular 4-helix bundle that is conferred with LDLR recognition ability after undergoing a lipid binding-induced conformational change. To investigate the structural basis for this phenomenon, site-directed spin label electron paramagnetic resonance spectroscopy experiments were conducted, focusing on the region near the C-terminal end of helix 4 (Ala-164). Using C112S apoE-N terminal as template, a series of single cysteine substitution variants (at sequence positions 161, 165, 169, 173, 176, and 181) were produced, isolated, and labeled with the nitroxide probe, methane thiosulfonate. Electron paramagnetic resonance analysis revealed that lipid association induced fixed secondary structure in a region of the molecule known to exist as random coil in the lipid free state. In a complementary approach, site-directed fluorescence analysis using an environmentally sensitive probe indicated that the lipid-induced transition of this region of the protein to alpha helix was accompanied by relocation to a more hydrophobic environment. In studies with full-length apoE single Cys variants, a similar random coil to stable backbone transition was observed, consistent with the concept that lipid interaction induced an extension of helix 4 beyond the boundary defining its lipid-free conformation. This structural transition likely represents a key conformational change necessary for manifestation of the LDLR recognition properties of apoE. PMID- 17079230 TI - The platelet-derived growth factor receptor alpha is destabilized by geldanamycins in cancer cells. AB - The heat shock protein HSP90 serves as a chaperone for receptor protein kinases, steroid receptors, and other intracellular signaling molecules. Targeting HSP90 with ansamycin antibiotics disrupts the normal processing of clients of the HSP90 complex. The platelet-derived growth factor receptor alpha (PDGFRalpha) is a tyrosine kinase receptor up-regulated and activated in several malignancies. Here we show that the PDGFRalpha forms a complex with HSP90 and the co-chaperone cdc37 in ovarian, glioblastoma, and lung cancer cells. Treatment of cancer cell lines expressing the PDGFRalpha with the HSP90 inhibitor 17-allylamino-17 demethoxygeldanamycin (17-AAG) promotes degradation of the receptor. Likewise, phospho-Akt, a downstream target, is degraded after treatment with 17-AAG. In contrast, PDGFRalpha expression is not affected by 17-AAG in normal human smooth muscle cells or 3T3 fibroblasts. PDGFRalpha degradation by 17-AAG is inhibited by the proteasome inhibitor MG132. High molecular weight, ubiquitinated forms of the receptor are detected in cells treated with 17-AAG and MG132. Degradation of the receptor is also inhibited by a specific neutralizing antibody to the PDGFRalpha but not by a neutralizing antibody to PDGF or by imatinib mesylate (Gleevec). Ultimately, PDGFRalpha-mediated cell proliferation is inhibited by 17-AAG. These results show that 17-AAG promotes PDGFRalpha degradation selectively in transformed cells. Thus, not only mutated tyrosine kinases but also overexpressed receptors in cancer cells can be targeted by 17-AAG. PMID- 17079231 TI - Down-regulation of manganese-superoxide dismutase through phosphorylation of FOXO3a by Akt in explanted vascular smooth muscle cells from old rats. AB - Manganese-superoxide dismutase (MnSOD) is one of the major cellular antioxidant defense systems. To study the effect of age on the regulation of MnSOD in the vasculature, we compared MnSOD expression and its transcriptional regulation in explanted vascular smooth muscle cells (VSMC) isolated from old (24 months old) versus young (6 months old) rats and grown in a normal (5 mM) or high (12.5 and 25 mM) glucose or tumor necrosis factor alpha (5 ng/ml) environment to induce oxidative stress. Both MnSOD protein and activity were reduced in VSMC from old compared with young animals. FOXO3a, a member of the family of Forkhead transcription factors, interacted with the promoter of the rat MnSOD gene at a specific binding site. Inhibition of FOXO3a transcription with small interfering RNA led to a reduction in MnSOD gene expression. VSMC from old rats had increased phosphorylated FOXO3a at Ser(253), which paralleled the reduction of MnSOD protein. Treatment of VSMC with 5 nm insulin-like growth factor-1 induced phosphorylation of Akt and FOXO3a over time, repressing FOXO3a DNA binding and consequently MnSOD gene expression. Furthermore, Akt activity was selectively increased in VSMC from the old, supporting the hypothesis that increased age related Akt activity might be responsible for the phosphorylation and inactivation of FOXO3a, which in turn down-regulates MnSOD transcription. PMID- 17079232 TI - Doxorubicin down-regulates Kruppel-associated box domain-associated protein 1 sumoylation that relieves its transcription repression on p21WAF1/CIP1 in breast cancer MCF-7 cells. AB - The role of post-translational modification, such as sumoylation, in modulating the efficacy of doxorubicin (Dox) treatment remains unclear. Transcriptional cofactor KRAB domain-associated protein 1 (KAP1) has been shown to complex with the KRAB zinc finger protein, ZBRK1, to repress the transcription of target genes. Through a combination of proteomic screening and site-directed mutagenesis approaches, we have identified lysines 554, 779, and 804 as the major sumoylation sites in KAP1. We then present evidence that Dox-mediated induction of cell cycle regulator p21 expression is differentially regulated by KAP1 sumoylation status. Moreover, the KAP1 sumoylation level was transiently decreased upon Dox exposure, and transfection with the KAP1 sumoylation mimetic, SUMO-1-KAP1, desensitizes breast cancer MCF-7 cells to Dox-elicited cell death. The sumoylation-dependent stimulation of KAP1 function is achieved by enhancing the methylation of H3-K9 and attenuating the acetylation of H3-K9 and H3-K14 at the p21 core promoter. We also show that occupancy of ZBRK1 response elements located at the p21 promoter by ZBRK1.KAP1 is independent of KAP1 sumoylation. Hence, sumoylation of KAP1 represses p21 transcription via a chromatin-silencing process without affecting interaction between KAP1.ZBRK1 and DNA, thus providing a novel mechanistic basis for the understanding of Dox-induced de-repression of p21 transcription. Taken together, our results suggest that Dox-induced decrease in KAP1 sumoylation is essential for Dox to induce p21 expression and subsequent cell growth inhibition in MCF-7 cells. PMID- 17079233 TI - Contacts between extracellular loop two and transmembrane helix six determine basal activity of the thyroid-stimulating hormone receptor. AB - A number of alanine mutations in extracellular loop two (ECL2) of the thyroid stimulating hormone receptor (TSHR) were found to increase or decrease basal activity when compared with the wild type receptor. K565A was identified as a mutant with decreased basal activity, and strongly impaired hormone induced signaling activity. To gain insights into how ECL2 mutants affect basal activity, we focused on constitutively activating pathogenic mutant I568V in ECL2, which exhibits elevated basal activity. Because our molecular model suggests that Ile 568 is embedded in an environment of hydrophobic residues provided by transmembrane helix bundle, we tested mutants in this region to identify potential interaction partner(s) for Ile-568. Indeed, the double mutant I568V/I640L (ECL2/TMH6) suppresses the increased basal activity exhibited by I568V alone. We suggest a spatial and functional relationship between ECL2 and TMH6 in which side chain interaction between Ile-568 and Ile-640 constrains the receptor in a conformation with low basal activity. Although the single mutant I640L exhibits basal activity lower than wild type, its differently branched and bulkier side chain complements the reduced side chain bulk in I568V, restoring wild type basal activity to the double mutant. This scenario is confirmed by the reciprocal double mutant I640V/I568L. The combination of basally increased activity of I640V and basally decreased activity of mutant I568L also restores basal activity of wild type TSHR. These and other mutant phenotypes reported here support a dynamic interface between TMH6 and ECL2. Disruption of this critical interface for signaling by introduction of mutations in TSHR can either increase or decrease basal activity. PMID- 17079234 TI - Cryoelectron microscopy and EPR analysis of engineered symmetric and polydisperse Hsp16.5 assemblies reveals determinants of polydispersity and substrate binding. AB - We have identified sequence and structural determinants of oligomer size, symmetry, and polydispersity in the small heat shock protein super family. Using an insertion mutagenesis strategy that mimics evolutionary sequence divergence, we induced the ordered oligomer of Methanococcus jannaschii Hsp16.5 to transition to either expanded symmetric or polydisperse assemblies. A hybrid approach combining spin labeling EPR and cryoelectron microscopy imaging at 10A resolution reveals that the underlying plasticity is mediated by a packing interface with minimal contacts and a flexible C-terminal tether between dimers. Twenty-four dimeric building blocks related by octahedral symmetry assemble into the expanded symmetric oligomer. In contrast, the polydisperse variant has an ordered dimeric building block that heterogeneously packs to yield oligomers of various sizes. Increased exposure of the N-terminal region in the Hsp16.5 variants correlates with enhanced binding to destabilized mutants of T4 lysozyme, whereas deletion of this region reduces binding. Transition to larger intermediates with enhanced substrate binding capacity has been observed in other small heat shock proteins including lens alpha-crystallin mutants linked to congenital cataract. Together, these results provide a mechanistic perspective on substrate recognition and binding by the small heat shock protein superfamily. PMID- 17079235 TI - Nanostructures of APOBEC3G support a hierarchical assembly model of high molecular mass ribonucleoprotein particles from dimeric subunits. AB - Human APOBEC3G (hA3G) is a cytidine deaminase that restricts human immunodeficiency virus (HIV)-1 infection in a vif (the virion infectivity factor from HIV)-dependent manner. hA3G from HIV-permissive activated CD4+ T-cells exists as an inactive, high molecular mass (HMM) complex that can be transformed in vitro into an active, low molecular mass (LMM) variant comparable with that of HIV-non-permissive CD4+ T-cells. Here we present low resolution structures of hA3G in HMM and LMM forms determined by small angle x-ray scattering and advanced shape reconstruction methods. The results show that LMM particles have an extended shape, dissimilar to known cytidine deaminases, featuring novel tail-to tail dimerization. Shape analysis of LMM and HMM structures revealed how symmetric association of dimers could lead to minimal HMM variants. These observations imply that the disruption of cellular HMM particles may require regulation of protein-RNA, as well as protein-protein interactions, which has implications for therapeutic development. PMID- 17079236 TI - Molecular architecture of the glucose 1-phosphate site in ADP-glucose pyrophosphorylases. AB - ADP-Glc pyrophosphorylase (PPase), a key regulatory enzyme in the biosynthetic pathway of starch and bacterial glycogen, catalyzes the synthesis of ADP-Glc from Glc-1-P and ATP. A homology model of the three-dimensional structure of the Escherichia coli enzyme complexed with ADP-Glc has been generated to study the substrate-binding site in detail. A set of amino acids in the model has been identified to be in close proximity to the glucose moiety of the ADP-Glc ligand. The role of these amino acids (Glu(194), Ser(212), Tyr(216), Asp(239), Phe(240), Trp(274), and Asp(276)) was studied by site-directed mutagenesis through the characterization of the kinetic properties and thermal stability of the designed mutants. All purified alanine mutants had 1 or 2 orders of magnitude lower apparent affinity for Glc-1-P compared with the wild type, indicating that the selected set of amino acids plays an important role in their interaction with the substrate. These amino acids, which are conserved within the ADP-Glc PPase family, were replaced with other residues to investigate the effect of size, hydrophobicity, polarity, aromaticity, or charge on the affinity for Glc-1-P. In this study, the architecture of the Glc-1-P-binding site is characterized. The model overlaps with the Glc-1-P site of other PPases such as Pseudomonas aeruginosa dTDP-Glc PPase and Salmonella typhi CDP-Glc PPase. Therefore, the data reported here may have implications for other members of the nucleotide diphosphoglucose PPase family. PMID- 17079237 TI - Minocycline delays but does not attenuate the course of experimental autoimmune encephalomyelitis in Streptococcus pneumoniae-infected mice. AB - OBJECTIVES: Experimental autoimmune encephalomyelitis (EAE), the animal model of multiple sclerosis (MS), can be aggravated by a mild Streptococcus pneumoniae infection. This study was performed to assess whether treatment with antibiotics inhibiting bacterial protein synthesis reduces the detrimental effect of infection on the course of EAE. METHODS: In vitro, release of proinflammatory pneumococcal products was studied by enzyme immunoassay and western blot. Seven days after induction of EAE (prior to the onset of symptoms) mice were infected intraperitoneally with S. pneumoniae and treated either with the inhibitors of bacterial protein synthesis minocycline or rifampicin, or with the beta-lactam ceftriaxone. RESULTS: During bacterial killing in vitro, minocycline and rifampicin released lower quantities of proinflammatory bacterial products from S. pneumoniae than ceftriaxone. Mice treated with minocycline developed symptoms of EAE 1 day later than mice treated with ceftriaxone. Neither minocycline nor rifampicin therapy, however, reduced the severity of EAE in comparison with ceftriaxone treatment. CONCLUSIONS: Although statistically significant (P = 0.04), a delay of 1 day in the onset of symptoms of EAE after minocycline treatment is of minor clinical relevance. These data do not support the hypothesis of superiority of a bacterial protein synthesis inhibitor over a beta lactam antibiotic for the treatment of concomitant infections during the latent phase of EAE or MS. PMID- 17079238 TI - Pharmacodynamic activity of ertapenem versus penicillin-susceptible and penicillin-non-susceptible Streptococcus pneumoniae using an in vitro model. AB - BACKGROUND: Ertapenem is a novel carbapenem with activity against both penicillin susceptible (MIC < or = 0.06 mg/L) and penicillin-non-susceptible (MIC > or = 0.12 mg/L) Streptococcus pneumoniae. This study assessed the pharmacodynamic activity of ertapenem against penicillin-susceptible and penicillin-non susceptible S. pneumoniae using an in vitro pharmacodynamic model. METHODS: Fifteen S. pneumoniae strains including 3 penicillin-susceptible and 12 penicillin-non-susceptible [4 penicillin-intermediate (MIC 0.12-1 mg/L) and 8 penicillin-resistant (MIC > or = 2 mg/L); with different resistance phenotypes including erythromycin-resistant (MIC > or = 1 mg/L), ciprofloxacin-resistant (MIC > or = 4 mg/L) and doxycycline-resistant (MIC > or = 8 mg/L)] were studied. The in vitro pharmacodynamic model was inoculated with 1 x 10(6) cfu/mL and ertapenem was dosed once daily at 0 and 24 h to simulate f (free) Cmax and t(1/2) obtained after a standard 1 g intravenous once daily dose in healthy volunteers (fCmax 15 mg/L, t(1/2) 4 h). Sampling was performed for 48 h to assess viable growth. RESULTS: Ertapenem T(> MIC) > or = 80% (ertapenem MICs < or = 0.5 mg/L) resulted in bactericidal (> or = 3 log10 killing) activity at 12, 24 and 48 h with complete eradication of penicillin-susceptible and penicillin-non susceptible S. pneumoniae from the model with no regrowth over the 48 h study period. Ertapenem T(> MIC) < or = 63% (ertapenem MIC > or = 1 mg/L) resulted in bactericidal activity at 12 h with regrowth at 24 and 48 h. The observed MICs for S. pneumoniae of ertapenem studied in the in vitro model did not change during the 48 h period, even for strains where regrowth occurred. CONCLUSIONS: Ertapenem is bactericidal against both penicillin-susceptible and penicillin-non susceptible S. pneumoniae (ertapenem MICs < or = 0.5 mg/L) when simulating free drug after 1 g intravenous once daily dosing. PMID- 17079239 TI - Are laboratory-based antibiograms reliable to guide the selection of empirical antimicrobial treatment in patients with hospital-acquired infections? AB - OBJECTIVES: Antibiograms are often taken into account to define a rational selection of an empirical antimicrobial therapy for treating patients with hospital-acquired infections. In this study, we performed a paired comparison between the antibiogram constructed with laboratory-based data and that formed with data subjected to prior clinical validation. METHODS: Between 2003 and 2005, the laboratory of microbiology printed in duplicate every individual susceptibility report corresponding to hospitalized patients and the copy was sent to the department of infection control. Every individual report was assessed in real time at the bedside of the patient by a multidisciplinary team for clinical significance and appropriateness of the specimen, as well as for the type, source and origin of the infection. Cumulative resistance rates were estimated in parallel at the laboratory with the whole data, and at the infection control department with data subjected to prior clinical validation. These rates were designated as 'laboratory-based' and 'clinically based', respectively. RESULTS: A total of 2305 individual susceptibility reports were assessed. Only 1429 (62.0%) were considered as clinically significant by the multidisciplinary team. Escherichia coli, Enterobacter cloacae, Citrobacter freundii group, Klebsiella species and Proteus mirabilis resistant to broad-spectrum cephalosporins, as well as methicillin-resistant Staphylococcus aureus, were significantly more frequent in the clinically based rates (P < or = 0.03). CONCLUSIONS: Laboratory-based data underestimate the frequency of several major resistant organisms in patients with hospital-acquired infection. Previous clinical validation of the individual susceptibility reports seems to be a suitable strategy to get more reliable data. PMID- 17079240 TI - Effect of caspofungin on trophozoites and cysts of three species of Acanthamoeba. AB - OBJECTIVES: Amoebic keratitis is difficult to treat, without total efficacy in some patients because of cysts that are less susceptible than trophozoites to the usual treatments. We investigated here the in vitro effectiveness of caspofungin, a new antifungal, against three species of Acanthamoeba. METHODS: Trophozoites and cysts of Acanthamoeba castellanii, Acanthamoeba culbertsoni and Acanthamoeba polyphaga were incubated with caspofungin at concentrations varying from 16 to 500 mg/L. RESULTS: The trophozoites of the three tested species were susceptible in vitro to caspofungin at a concentration of 250 mg/L (206 microM). Furthermore, this drug was cysticidal at a concentration of 500 mg/L (412 microM) against A. castellanii and A. culbertsoni. CONCLUSIONS: Caspofungin could represent, if in vivo studies confirm its efficacy, a new anti-Acanthamoeba compound. PMID- 17079241 TI - Proinflammatory activation of Toll-like receptor-2 during exposure of penicillin resistant Streptococcus pneumoniae to beta-lactam antibiotics. AB - OBJECTIVES: Disease caused by penicillin-resistant Streptococcus pneumoniae (PRSP) is associated with more suppurative complications than disease caused by penicillin-susceptible S. pneumoniae (PSSP). Exposure of S. pneumoniae to beta lactam antibiotics enhances the proinflammatory activation of human cells by pneumococci via Toll-like receptor-2 (TLR2). To test the hypothesis that penicillin resistance influences cellular TLR2 activation by beta-lactam-exposed pneumococci, we compared TLR2 induction by PSSP (MIC 0.06 mg/L) and a high-level PRSP clinical isolate (159; MIC 16 mg/L) following exposure to penicillin and cefotaxime. METHODS: Both organisms were treated with penicillin or cefotaxime at and around the MIC. TLR2 signalling was measured as relative IL-8 promoter activation in transfected HeLa cells. RESULTS: On exposure to penicillin, log phase PSSP and PRSP induced TLR2-proinflammatory activation at levels significantly higher than unexposed bacteria, and maximal in each case at the MIC. Transformants containing low-affinity penicillin-binding proteins (PBP) 2x, 1a and 2b exhibited stepwise resistance to cefotaxime and penicillin. TLR2 activation following penicillin treatment was dependent on an abnormal cell wall (PBP1a and 2x) and autolysis (PBP2b). High affinity PBP2x was required for this effect to be observed in log-phase pneumococci exposed to cefotaxime at the MIC. Cefotaxime-mediated TLR2 activation was not observed in lag-phase transformants exposed to sub-lethal concentrations. CONCLUSIONS: These data show that PRSP have similar TLR2-proinflammatory effects to PSSP when exposed to beta-lactam antibiotics but the antibiotic concentration relative to the MIC is critical. This has implications for treatment of pneumococcal disease when tissue concentrations of antibiotic are close to the MIC. PMID- 17079242 TI - Antisense peptide-phosphorodiamidate morpholino oligomer conjugate: dose-response in mice infected with Escherichia coli. AB - OBJECTIVES: Phosphorodiamidate morpholino oligomers (PMOs) are DNA analogues that inhibit translation by an antisense mechanism. Membrane-penetrating peptides attached to PMOs increase PMO efficacy by enhancing penetration through bacterial membranes. The objectives of these experiments are to demonstrate gene-specific efficacy and establish a dose-response relationship of a peptide-PMO conjugate. METHODS: An 11-base PMO (AcpP) targeted at acpP (an essential gene) of Escherichia coli was synthesized and conjugated with the cell-penetrating peptide RFFRFFRFFRXB (X is 6-aminohexanoic acid and B is beta-alanine). Mice were infected by intraperitoneal (i.p.) injection with K-12 E. coli W3110, and treated i.p. at 15 min and 12 h post-infection with various amounts of AcpP peptide-PMO conjugate, AcpP PMO without attached peptide, scrambled base sequence PMOs or ampicillin. A strain (LT1) of E. coli was constructed by replacing acpP with an allele that has four wobble base substitutions in the region targeted by the PMO. RESULTS: Twelve hours after a single treatment, 30 microg of AcpP peptide-PMO or 3 mg of AcpP PMO reduced bacteraemia by 3 orders of magnitude compared with treatment with water. Neither scrambled base sequence PMO controls nor 30 microg of ampicillin reduced bacteraemia. Two treatments with 30 microg of AcpP peptide PMO reduced cfu significantly more than four treatments with 15 microg at 15 min, 4, 8 and 12 h. Mice treated with doses of AcpP peptide-PMO > 30 microg showed further reductions in plasma cfu. Survival 48 h after treatment with 2 x 30 microg (3 mg/kg) of AcpP peptide-PMO or 2 x 3 mg (300 mg/kg) of AcpP PMO was 100%, compared with 20% for mice treated with water or scrambled base sequence PMO controls. However, survival was reduced to 75% and 0% for mice treated with 2 x 300 microg and 2 x 1 mg of AcpP peptide-PMO, respectively. A conjugate made from the D-isomeric form of each amino acid was less effective than the L-amino acid equivalent, and required 2 x 300 microg treatments for significant reduction in bacteria and survival. Mice infected with LT1 and treated with AcpP peptide PMO did not survive and had the same amount of bacteria in the blood as mice treated with water, whereas those treated with 2 x 100 microg of AcpPmut4 peptide PMO (complementary to the mutated allele) survived, and had a 3 orders of magnitude reduction in bacteria in the blood at 24 h post-infection. CONCLUSIONS: Both AcpP peptide-PMO and AcpP PMO significantly reduced bacteraemia and promoted survival of mice infected with E. coli W3110. The conjugate was about 50-100 times more potent than the PMO without attached peptide. The L-isomeric peptide PMO was 10 times more potent than the D-isomeric equivalent. The conjugate apparently was toxic at doses > or = 2 x 300 microg/mouse (30 mg/kg). PMOs produced a sequence-specific antibiotic effect and the conjugate had a therapeutic index (toxic dose/effective dose) approximately equal to 10 in a mouse model of infection. PMID- 17079243 TI - Small-colony variants: a novel mechanism for triclosan resistance in methicillin resistant Staphylococcus aureus. AB - OBJECTIVES: A little-understood mode of antimicrobial resistance in Staphylococcus aureus is the evolution of a sub-population of small-colony variants (SCVs). SCVs are a cause of persistent and recurring infections refractory to antimicrobial chemotherapy. Following the inadvertent isolation of suspected SCVs growing in the presence of triclosan we set out to evaluate the formation of these colonial mutants and assess their antimicrobial susceptibility. METHODS: SCVs were isolated on Mueller-Hinton agar supplemented with 1 mg/L triclosan. SCV formation frequency was calculated using a selection of both clinical methicillin-resistant S. aureus (MRSA) isolates and methicillin susceptible S. aureus strains. Antimicrobial susceptibility was assessed and the fabI gene of SCVs was sequenced to ensure resistance was not mediated by mutation of this gene. RESULTS: We have found in vitro that triclosan can select for S. aureus colonies showing the characteristic SCV phenotype with low-level triclosan resistance and which coincidently have reduced susceptibility to penicillin and gentamicin. Additionally, triclosan-isolated SCVs were shown to have an increased tolerance to the lethal effects of triclosan. CONCLUSIONS: We propose the formation of SCVs by S. aureus is a novel mechanism of resistance to low concentrations of triclosan, which for 25 years has been used widely in the domestic setting in various consumer healthcare products. More recently it has been recommended for the control of MRSA. Consequently, our results identify the potential for treatment failure in infections already notoriously difficult to eradicate. It remains unclear to what extent isolates with decreased susceptibility to triclosan would develop and have the fitness to survive under real world conditions. PMID- 17079244 TI - Vascular endothelial growth factor levels in serum and plasma from patients undergoing controlled ovarian hyperstimulation for IVF. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) has been investigated as a marker of ovarian response to controlled ovarian hyperstimulation and as a predictor of ovarian hyperstimulation syndrome (OHSS) in IVF cycles. In most studies, serum has been used for circulating VEGF concentration measurement, but it has been suggested that plasma is the preferred medium to measure VEGF levels because of the potential contribution of VEGF released from platelets during blood clotting. This study investigated VEGF concentrations in paired serum and plasma samples from patients undergoing controlled ovarian hyperstimulation for IVF. METHODS: Serum and plasma VEGF levels, as well as the number of platelets, were measured in 30 IVF patients who comprised three study groups delineated according to the estradiol (E(2)) serum concentration reached on the day of HCG administration: 10 patients having low E(2) serum levels (<1500 pg/ml, group L), 10 patients having intermediate E(2) serum levels (1500-3000 pg/ml, group I) and 10 patients having high E(2) serum levels (>3000 pg/ml, group H). RESULTS: There was a statistically significant correlation between plasma and serum VEGF levels (rho = 0.61; P < 0.005) for the entire population studied, although serum values were higher by a factor of approximately 6-fold. No significant correlation was found between peripheral blood VEGF concentrations and serum E(2) or follicle number on HCG day or the number of oocytes collected. Similarly, paired serum and plasma VEGF measurements did not correlate with platelet count. CONCLUSIONS: Serum and plasma VEGF concentrations are strongly correlated in paired samples from infertile patients undergoing controlled ovarian hyperstimulation. However, neither serum nor plasma VEGF levels were correlated with parameters associated with ovarian follicular activity. Peripheral blood VEGF levels were not correlated with platelet count. PMID- 17079245 TI - Oocyte donation for stem cell research. AB - The future success of stem cell research by means of somatic cell nuclear transfer (SCNT) depends on a sufficient supply of human oocytes. However, oocyte donation presents certain risks for the donor, and concerns for women's welfare are rightly vocalized. At the same time, these risks are comparable with the risks faced by other healthy research subjects. Thus, research donation can withstand ethical scrutiny if it fulfils the same conditions as other research involving healthy human subjects. Specifically, this means that the benefits of the research project need to outweigh the harms, that risks must be minimized, that informed consent has to be guaranteed by averting undue inducement and the recruitment of vulnerable women and that donors can and should be reimbursed for their research participation. PMID- 17079246 TI - A new multiparameter flow cytometric method for human semen analysis. AB - BACKGROUND: The objectives of this study were (i) to evaluate whether the combined use of Syto 16 and 7-amino-actinomycin-D (7-AAD) allows the detection of sperm apoptosis and (ii) to describe a new multiparameter flow cytometric method to assess simultaneously sperm concentration (SC), viability and apoptosis as well as leukocyte concentration. METHODS: Semen samples from 68 patients were evaluated according to World Health Organization (WHO) criteria (normal, n=26; abnormal, n=42). The detection of activated caspases before and after betulinic acid (BA) incubation was carried out in 13 semen samples by flow cytometry using fluorescein-labelled inhibitors of caspases (FLICA). A multiparameter flow cytometric analysis was performed in 55 semen samples. Fluorescent microspheres were used to assess SC. Sperm apoptosis was detected by staining sperm with Syto 16 and 7-AAD. Leukocytes were counted using monoclonal anti-CD45. RESULTS: A significant correlation between the percentage of the spermatozoa with low Syto 16 fluorescence and the percentage of spermatozoa containing activated caspases was found (r=0.68, P=0.0106; n=13). After incubation with BA, an increase of the percentage of apoptotic cells was observed in all samples, using both the Syto 16/7-AAD and the caspase activation methods. There was a good correlation between flow cytometry and optical microscopy for sperm (r=0.98, P < 0.0001) and leukocyte counting (r=0.64, P <0.0001). The percentage of apoptotic sperm was inversely correlated with both SC (r=-0.303, P=0.0246) and morphology (r=-0.384, P=0.0050) but not with motility. CONCLUSIONS: The combination of Syto 16/7-AAD provides a sensitive assay to detect sperm apoptosis. The multiparameter flow cytometric method described offers the possibility of a simultaneous, simple, rapid and accurate assessment of several semen parameters. PMID- 17079247 TI - Cardiac dysfunction in patients with systemic lupus erythematosus and antiphospholipid syndrome. AB - OBJECTIVE: To comparatively assess the parameters of systolic and diastolic cardiac function in patients with systemic lupus erythematosus (SLE) and antiphospholipid syndrome (APS). METHODS: Consecutive patients (n=74) who were free of cardiovascular symptoms were divided into four groups: (1) SLE (n=23); (2) SLE with antiphospholipid antibodies (aPL; n=18); (3) SLE with APS (n=20); and (4) primary antiphospholipid syndrome (PAPS; n=13). Pulsed, continuous, colour Doppler echocardiography, and M-mode and B-mode studies were performed. RESULTS: Left ventricular end diastolic and end systolic dimensions were higher in SLE as compared with patients with PAPS (p=0.022 and 0.022, respectively), with a trend towards a lower fractional shortening in SLE (p=0.07), suggesting systolic dysfunction. Parameters of diastolic function were more impaired in patients with APS, reflected by lower left ventricular and right ventricular E wave to A wave (E:A) ratios in patients with APS (groups 3, 4) compared with those without APS (groups 1, 2; 1.15 (0.40) v 1.49 (0.43), p=0.001 and 1.19 (0.31) v 1.49 (0.41), p=0.001, respectively) and a more prolonged left ventricular isovolumic relaxation time (IVRT; 94.2 (24.6) v 84.4 (17) ms, respectively, p=0.055). Patients with APS were older than those without APS (47.12 (14.86) v 34.29 (12.6), p=0.0001). Patients with SLE were younger than those with PAPS (38.19 (14.68) v 48.53 (13.97), p=0.023). CONCLUSION: Abnormal echocardiographic findings were detected frequently in asymptomatic patients with SLE or PAPS. Although patients with SLE were younger, left ventricular systolic function was more impaired in patients with SLE compared with those with PAPS, whereas left ventricular and right ventricular diastolic function, as reflected by IVRT and E:A ratios, were significantly more impaired in patients with APS. PMID- 17079249 TI - Personalised care, access, quality and team coordination are the main dimensions of family medicine output. AB - BACKGROUND: Health organisations continually seek good output indicators of family medicine health care provision because they are accountable to society, they need to compare services, and need to evaluate the impact of organisational reforms. OBJECTIVES: Using the sources of information routinely available in health-service management, we sought to assess the groups of components of primary health care output that best serve to define the outcome of family medicine services. DESIGN: Cross-sectional descriptive study. SITE: Primary health care in Catalunya. PARTICIPANTS: 213 primary health care teams. MEASUREMENTS: Information was collected on team structure, user satisfaction, quality-of-professional-life of the health care professionals, and physicians' drug prescription. Confirmatory Factor Analysis was used to assess the number of dimensions that best explained the family medicine outcome. RESULTS: The model that best fits the structure of the data (AGFI=0.778) is that which consists of three dimensions i.e. (1) the individual accessibility to the services and professional-patient relationship; (2) the coordination within the health care team; (3) the scientific-technical quality of the service. The first two of these dimensions were correlated between themselves, but the third was totally independent of the other two. CONCLUSIONS: Using sources of information that are routinely employed in primary health care services management, the model enables the measurement of the output of family medicine by considering the dimensions such as inter-personnel relationships, internal coordination of the team and the scientific-technical quality of the service. Despite its simplicity, this measure of the output incorporates the views not only of the users but of the health care professionals, as well. PMID- 17079248 TI - High IgA rheumatoid factor levels are associated with poor clinical response to tumour necrosis factor alpha inhibitors in rheumatoid arthritis. AB - OBJECTIVE: To investigate whether rheumatoid factor isotypes and anti-cyclic citrullinated peptide (anti-CCP) antibodies are related to clinical response in patients with rheumatoid arthritis treated with tumour necrosis factor alpha (TNFalpha) inhibitors. METHODS: The study was carried out on 132 patients with advanced rheumatoid arthritis refractory to disease-modifying antirheumatic drugs. Patients were treated with infliximab (n = 63), etanercept (n = 35) or adalimumab (n = 34). All patients completed 1 year of follow-up, and 126 were evaluable for clinical response according to the disease activity score (DAS) criteria. IgM, IgA and IgG rheumatoid factors and anti-CCP antibodies were assessed by ELISA both before anti-TNFalpha treatment and 1 year later. RESULTS: The DAS response was reached in 66% of evaluable patients (61% infliximab, 65% etanercept and 76% adalimumab; p = 0.354). A significant reduction in the rheumatoid factor level was reported by all treatment groups after 1 year. The frequency of positive tests for the different antibodies did not differ between responders and non-responders at baseline; however, significantly higher IgA rheumatoid factor levels were reported by the non-responder group (130.4 U/ml (interquartile range 13.8-276.7) v 24.8 U/ml (10.2-90.8); p = 0.003). A significant decrease (p<0.001) in the levels of all rheumatoid factor isotypes in the responder group was reported after 1 year of treatment, whereas anti-CCP antibody levels were not significantly affected. CONCLUSIONS: According to the clinical response, anti-TNFalpha agents seem to reduce IgM, IgG and IgA rheumatoid factor levels. More interestingly, high pretreatment levels of IgA rheumatoid factor are associated with a poor clinical response to TNFalpha inhibitors. PMID- 17079250 TI - Logistic support service improves processes and outcomes of diabetes care in general practice. AB - BACKGROUND: Guidelines for type 2 diabetes care in general practice are well known and accepted, but the implementation falls short. OBJECTIVE: To implement these guidelines by introducing a diabetes support service (DSS) to support the care delivered by the GP. METHODS: A controlled, non-randomised study with delayed intervention in the control group; 78 GPs (n=51 for the intervention and n=21 for the control group) in the south of the Netherlands and 613 of their type 2 diabetic patients participated. Data were collected on the frequency, content and results of the check-ups (fasting blood glucose, HbA1c, cholesterol, cholesterol/HDL ratio, triglycerides, creatinine, blood pressure, fundus photography, foot examination, body mass index and smoking status) for 3 years. The year before signing up with the DSS was taken for the pre-measurements and after 2 years of DSS the post-measurements took place. The effect of the DSS was analysed in a mixed model with repeated measurement covariance structure. RESULTS: At baseline the intervention and control group did not differ in control frequency and outcome (HbA1c). After the intervention the percentage of patients that attended four or more quarterly check-ups (with at least testing of fasting blood glucose or HbA1c) increased from 59 to 78%. In contrast, the frequency of check-ups in the control group remained constant. This effect was significant. The HbA1c remained the same in the intervention group while there was a significant deterioration in the HbA1c in the control group. CONCLUSION: Simple logistic support by the DSS proved to have the capacity to implement type 2 diabetes guidelines in general practice. PMID- 17079252 TI - Free-flow oxygen delivery to newly born infants. AB - Resuscitation guidelines recommend administration of free-flow oxygen to newly born infants who breathe but remain cyanosed. Self-inflating resuscitation bags are described as unreliable for this purpose. We measured oxygen concentrations >or=80% delivered through a 240 mL Laerdal self-inflating resuscitation bag and from 5 mm tubing inside a cupped hand. PMID- 17079251 TI - Phenocopies in BRCA1 and BRCA2 families: evidence for modifier genes and implications for screening. AB - BACKGROUND: The identification of BRCA1 and BRCA2 mutations in familial breast cancer kindreds allows genetic testing of at-risk relatives. Those who test negative are usually reassured and additional breast cancer surveillance is discontinued. However, we postulated that in high-risk families, such as those seen in clinical genetics centres, the risk of breast cancer might be influenced not only by the BRCA1/BRCA2 mutation but also by modifier genes. One manifestation of this would be the presence of phenocopies in BRCA1/BRCA2 kindreds. METHODS: 277 families with pathogenic BRCA1/BRCA2 mutations were reviewed and 28 breast cancer phenocopies identified. The relative risk of breast cancer in those testing negative was assessed using incidence rates from our cancer registry based on local population. RESULTS: Phenocopies constituted up to 24% of tests on women with breast cancer after the identification of the mutation in the proband. The standardised incidence ratio for women who tested negative for the BRCA1/BRCA2 family mutation was 5.3 for all relatives, 5.0 for all first degree relatives (FDRs) and 3.2 (95% confidence interval 2.0 to 4.9) for FDRs in whose family all other cases of breast and ovarian cancer could be explained by the identified mutation. 13 of 107 (12.1%) FDRs with breast cancer and no unexplained family history tested negative. CONCLUSION: In high-risk families, women who test negative for the familial BRCA1/BRCA2 mutation have an increased risk of breast cancer consistent with genetic modifiers. In light of this, such women should still be considered for continued surveillance. PMID- 17079253 TI - Noninvasive and invasive evaluation of pulmonary arterial pressure in highlanders. AB - The purpose of the present study was to evaluate Doppler echocardiography for the detection of pulmonary hypertension in high-altitude inhabitants. In total, 60 (55 male) patients aged 18-71 yrs were recruited from an ECG screening programme applied to 1,430 inhabitants living at an altitude of 2,500-3,600 m in Kyrgyzstan. Of these, 44 met ECG criteria for right ventricular hypertrophy. All underwent Doppler echocardiography followed by a cardiac catheterisation within 7 days of arrival in Bishkek (Kyrgyzstan; altitude 760 m). Pulmonary flow acceleration time and the maximum velocity of tricuspid regurgitation were measured. Sufficient quality tricuspid regurgitant jets were recovered in only 28% of the patients. Therefore, pulmonary artery pressure was estimated from the pulmonary flow acceleration time, which was recovered in 100% of the patients. It was found that 37 (62%) of the patients had pulmonary hypertension on echocardiography. Pulmonary hypertension was confirmed in 29 patients on catheterisation. Pulmonary hypertension was detected with 70% sensitivity and 88% specificity by echocardiography, as compared to 59% sensitivity and 81 % specificity by ECG. The correlation coefficient between echocardiography and catheterisation studies was r(2) = 0.78. It is concluded that a combination of ECG and echocardiography may be useful for screening high-altitude pulmonary hypertension. PMID- 17079254 TI - Lung and blood mononuclear cell responses of tuberculosis patients to mycobacterial proteins. AB - The differences in specificity of human lung and peripheral lymphocytes for mycobacterial antigens (Ag) need to be evaluated in order to identify vaccine candidates against pulmonary tuberculosis (TB). Therefore, the present study examined the response to low molecular weight secretory proteins of Mycobacterium tuberculosis in bronchoalveolar lavage (BAL) and peripheral blood mononuclear cells (PBMCs) from minimal pulmonary TB and non-TB patients. Ag85A, Ag85B, culture filtrate protein (CFP)-31, CFP-22.5, CFP-21, M. tuberculosis protein-64 and an as yet uncharacterised 19 kDa protein were found to be predominantly recognised by BAL cells of TB patients on the basis of lymphocyte proliferation and significant interferon-gamma release. However, recognition of CFP-8, 6-kDa early secreted antigenic target, CFP-10, CFP-14.5, M. tuberculosis secretory protein-17 and five other as yet uncharacterised low molecular weight polypeptides was found to be high on the basis of lymphocyte proliferation at the level of PBMCs. Furthermore, BAL macrophages, and not blood monocytes, were found to produce nitric oxide (NO) in response to mycobacterial Ags. Among polypeptides predominantly recognised by BAL lymphocytes, only Ag85A and Ag85B were found to induce both NO and interleukin-12 (p40) by alveolar macrophages. In conclusion, the present results indicate heterogeneity in antigen recognition by bronchoalveolar lavage cells and peripheral mononuclear blood cells of minimal tuberculosis patients, and also suggest the utility of antigen 85 complex polypeptides for the development of a future mucosal antituberculous vaccine. PMID- 17079255 TI - Exhaled breath condensate pH standardised for CO2 partial pressure. AB - Exhaled breath condensate (EBC) pH is considered to reflect the acid-base balance of the airways. Current pH measurements do not take into account the effect of CO2. The aim of the present study was to determine the effect of condensate CO2 partial pressure on pH and to provide a more precise mode of EBC pH determination. Condensate pH and CO2 partial pressure were measured in parallel from 12 healthy volunteers and 12 asthmatics using a blood gas analyser in neat, argon de-aerated and CO2-loaded samples. The regression analysis was used to test the relationship between pH and CO2, and to calculate the pH at a CO2 level of 5.33 kPa (physiological alveolar CO2 partial pressure). Reproducibility of different pH readings was compared using the Bland-Altman test. Condensate CO2 concentration was variable both in neat and argon de-aerated samples. There was a close negative logarithmic relationship between CO2 and pH. Calculation of pH at a CO2 level of 5.33 kPa provided reproducibility approximately six times as good as that of the currently used measurements. Condensate CO2 partial pressure influences pH measurements. Determination of pH at a standard CO2 level provides the most reproducible condensate pH values to date. PMID- 17079256 TI - Addition of sildenafil to bosentan monotherapy in pulmonary arterial hypertension. AB - Combination therapy has been recommended for the treatment of pulmonary arterial hypertension (PAH). However, there is scant information on combination therapy after failure of monotherapy, particularly in patients with scleroderma associated PAH (PAH-SSD). From a group of 82 consecutive patients with PAH who received initial bosentan monotherapy, a total of 13 idiopathic PAH (IPAH) and 12 PAH-SSD patients requiring additional therapy with sildenafil were studied. Sildenafil was added for clinical deterioration based upon symptoms, New York Heart Association (NYHA) classification or 6-min walk distance (6MWD). Clinical data and haemodynamics were collected at baseline. Assessments were made at 1-3 month intervals. At baseline, there were no differences in demographics, NYHA classification, haemodynamics or 6MWD between the two groups. After initiation of bosentan, both groups experienced clinical improvement but ultimately deteriorated (median time to monotherapy failure 792 versus 458 days for IPAH and PAH-SSD patients, respectively). After addition of sildenafil, more IPAH patients tended to improve in NYHA class (five out of 13 versus two out of 12) and walked further (mean difference in 6MWD 47+/-77 m versus -7+/-40 m) compared with PAH SSD patients. In conclusion, addition of sildenafil after bosentan monotherapy failure improved New York Heart Association class and 6-min walk distance in idiopathic pulmonary arterial hypertension patients but failed to improve either parameter in scleroderma-associated pulmonary arterial hypertension patients. Additional studies are needed to assess the tolerability and efficacy of this combination in patients with scleroderma-associated pulmonary arterial hypertension. PMID- 17079257 TI - Ultrasound-assisted transthoracic biopsy: fine-needle aspiration or cutting needle biopsy? AB - The present study compared the diagnostic yield of ultrasound-assisted cutting needle biopsy (CNB) and fine-needle aspiration biopsy (FNAB) in chest lesions. A physician performed ultrasound and FNAB with a 22-G spinal needle in all patients, directly followed by a 14-G CNB in patients without contraindication. A total of 155 consecutive lesions arising from the lung (74%), pleura (12%), mediastinum (11%) or chest wall (3%) in patients with a final diagnosis of lung carcinoma (74%), other malignant tumours (12%), non-neoplastic disease (9%) or unknown (5%) were prospectively included. The overall diagnostic yield was 87%. Combined specimens were obtained in 123 lesions (79%). In these, yields of FNAB, CNB and both methods combined were 82, 76 and 89%, respectively. FNAB was significantly better than CNB in lung carcinoma (95 versus 81%) but CNB was superior in noncarcinomatous tumours and in benign lesions. On-site cytology was 90% sensitive and 100% specific for predicting a positive FNAB. One patient required drainage for pneumothorax (0.6%). Ultrasound-assisted fine-needle aspiration biopsy performed by chest physicians is an accurate and safe initial diagnostic procedure in patients with a high clinical probability of lung carcinoma. All other patients should undergo concurrent fine-needle aspiration biopsy and cutting-needle biopsy. PMID- 17079258 TI - Eotaxin-2 in sputum cell culture to evaluate asthma inflammation. AB - The aim of the present study was to elucidate whether the culture of cells recovered from induced sputum may represent a suitable model to evaluate cytokine and chemokine production by airway inflammatory cells. Sputum induction was performed in 21 normal subjects and 30 asthmatic patients. A total of 21 out of the 30 asthmatic patients were taking inhaled corticosteroids, while the remaining nine were steroid-naive asthmatics. The steroid-naive group was evaluated before and after a 14-day treatment with oral prednisone (40 mg.day( 1)). The supernatant of lysed and centrifuged sputum and the supernatant of sputum cell culture were analysed. Tumour necrosis factor-alpha, interleukin (IL) 8 (CXCL8), IL-1beta, IL-13 and eotaxin-2 (CCL24) concentrations were determined by specific ELISA. Eotaxin-2 production by cell culture was higher in the asthma group (131+/-108 pg.mL(-1)) than in the control group (36+/-41 pg.mL(-1)) and treatment with oral corticosteroids eliminated this difference. In addition, reduction of eotaxin-2 levels by corticosteroid treatment was greater in cell culture (81.3% reduction) than in sputum (26.4%). There was correlation between the decrease in eotaxin-2 production and the decrease in blood eosinophil number and between eotaxin-2 and eosinophils in sputum. Eotaxin-2 may play an important role in asthma and the response to corticosteroid treatment suggests that analysis of sputum cell culture is relevant as an inflammatory parameter. PMID- 17079259 TI - Impact of blood transfusions on outcome after pneumonectomy for thoracic malignancies. AB - The aim of the present study was to determine the risk factors and impact on outcome of blood transfusions following pneumonectomy for thoracic malignancies. A retrospective analysis of 432 consecutive patients was carried out, of whom 183 (42.4%) were transfused post-operatively. The associations between blood transfusions and 20 variables were assessed by univariate and multivariate analysis. Survival analysis included log-rank test and Cox regression model. Patient age, neoadjuvant treatment, completion pneumonectomy and extended procedures were independent predictors of transfusion. It was found that 30-day mortality increased significantly from 2.4% (no transfusion) to 10.9 and 21.9% (2 red blood cell packs, respectively). Blood transfusion was the strongest predictor of 30-day mortality (odds ratio (OR) 10; 95% confidence interval (CI): 3.7-27), respiratory failure (OR 19.2; 95% CI 7.4-49.4) and infectious complications (OR 3; 95% CI 1.5-6.2). In the 367 lung cancer patients, a significantly lower 5-yr survival was observed in univariate analysis of transfused patients (27.8+/-5.4% versus 39.4+/-4.5%). In a Cox regression analysis, blood transfusion was no longer found to be significant. A dose-related correlation is suggested between blood transfusion and early mortality through an increase of infectious and respiratory complications. In contrast, blood transfusion had no independent adverse impact on long-term survival. PMID- 17079260 TI - Self-management in bronchiectasis: the patients' perspective. AB - Self-management programmes for chronic disease are a high priority for healthcare providers. The content and method of delivery of self-management should give consideration to the specific requirements of the disease population. The aims of the present study were to assess the physical and psychosocial impact of bronchiectasis, to determine whether patients with bronchiectasis are receptive to self-management and to identify any obstacles or sources of support for a disease-specific self-management programme. A total of 32 patients with a diagnosis of bronchiectasis attended four focus groups. Each focus group was videotaped and subjected to qualitative analysis using the grounded theory approach. Bronchiectasis had an impact on patients' physical and psychosocial well-being. Patients demonstrated the potential to self-manage with strategies including self-regulation of medication and airway clearance. Perceived obstacles to self-management included lack of information and confidence. Patients suggested that self-management could be promoted through disease-specific information and appropriate healthcare procedures. In summary, patients with bronchiectasis have their lives disrupted by this chronic condition, but are receptive to self-management. The present study has provided information from the patients' perspective of elements which need to be included in a successful disease-specific self-management programme. PMID- 17079261 TI - Genetic and environmental effects on exhaled nitric oxide and airway responsiveness in a population-based sample of twins. AB - Elevated levels of exhaled nitric oxide (eNO) and airway hyperresponsiveness are intermediate phenotypes of asthma. Using population-based data collected from a sample of twins, the present authors estimated the relative contribution of genes, family environment and nonshared environmental influences to variations in eNO and airway responsiveness (AR). In addition, the genetic and environmental sources of covariation between these two asthma-related phenotypes were investigated. The study population comprised a random sample of 377 adult twins identified through the Norwegian Twin Registry. The main outcome variables were eNO and AR to methacholine. Genetic effects accounted for 60% of the variation in eNO. Family environment accounted for 30% of the variation in AR, while nonshared environmental influences explained the remaining variation for both measures. For both eNO and AR, there were significant regression effects for atopy and smoking. The small, but significant association between eNO and AR was primarily explained by genetic factors. Sub-analyses restricted to atopic and nonsmoking twins strengthened the observation. In conclusion, variations in exhaled nitric oxide and airway responsiveness appear to be explained by different genetic and environmental variance structures. Variation in exhaled nitric oxide is explained by genetic and nonshared environmental effects, whereas an environmental model best explains the variation in airway responsiveness. Common genetic effects explain the small but significant association between exhaled nitric oxide and airway responsiveness. PMID- 17079262 TI - Expiratory flow limitation detected by forced oscillation and negative expiratory pressure. AB - The within-breath change in reactance (Delta(rs)) measured by forced oscillation technique (FOT) at 5 Hz reliably detects expiratory flow limitation in chronic obstructive pulmonary disease (COPD). The present study compared this approach to the standard negative expiratory pressure (NEP) method. In total, 21 COPD patients were studied by applying both techniques to the same breath and in 15 patients the measurements were repeated after bronchodilator. For each patient and condition five NEP tests were performed and independently scored by three operators unaware of the FOT results. In 180 tests, FOT classified 53.3% as flow limited. On average, the operators scored 27.6% of tests flow limited and 47.6% non-flow limited, but could not score 24.8%. The methods disagreed in 7.9% of cases; in 78% of these the NEP scores differed between operators. Bronchodilation reduced NEP and DeltaX(rs) scores, with only the latter achieving significance. Averaging the operators' NEP scores, a threshold between 24.6-30.8% of tidal volume being flow limited by NEP produced 94% agreement between methods. In conclusion, when negative expiratory pressure and forced oscillation technique were both available they showed good agreement. As forced oscillation technique is automatic and can measure multiple breaths over long periods, it is suitable for monitoring expiratory flow limitation continuously and identifying patients' breathing close to the onset of expiratory flow limitation, where intermittent sampling may be unrepresentative. PMID- 17079263 TI - Naphthoquinone enhances antigen-related airway inflammation in mice. AB - The current authors have previously demonstrated that diesel exhaust particles (DEP) enhance antigen-related airway inflammation in mice. Furthermore, a recent study has shown that organic chemicals in DEP, rather than their carbonaceous nuclei, are important contributors to the aggravating effects of airway inflammation. However, the components in DEP responsible for the enhancing effects on the model remain to be identified. The current authors investigated the effects of naphthoquinone (NQ), one of the extractable chemical compounds of DEP, on antigen-related airway inflammation, local expression of cytokine proteins, and antigen-specific immunoglobulin (Ig) production in mice. Pulmonary exposure to NQ dose-dependently aggravated antigen-related airway inflammation, as characterised by infiltration of eosinophils and lymphocytes around the airways and an increase in goblet cells in the bronchial epithelium. Combined exposure to NQ and antigen enhanced the local expression of interleukin (IL)-4, IL-5, eotaxin, macrophage chemoattractant protein-1 and keratinocyte chemoattractant, compared with exposure to antigen or NQ alone. Also, NQ exhibited adjuvant activity for the antigen-specific production of IgG(1) and IgG(2a). These results provide the first experimental evidence that naphthoquinone can enhance antigen-related airway inflammation in vivo, and that naphthoquinone can, to some extent, partly play a role in the pathogenesis of diesel exhaust particle toxicity on the condition. PMID- 17079264 TI - SUPPRESSOR OF FRI 4 encodes a nuclear-localized protein that is required for delayed flowering in winter-annual Arabidopsis. AB - The floral inhibitor FLOWERING LOCUS C (FLC) is a crucial regulator of flowering time in Arabidopsis, and is positively regulated by the FRIGIDA (FRI) gene in late-flowering winter-annual accessions. In rapid-cycling accessions, FLC expression is suppressed by the autonomous floral-promotion pathway (AP); thus AP mutants contain high levels of FLC and are late flowering. Previous work has shown that the upregulation of FLC in FRI- or AP-mutant backgrounds is correlated to an increase in histone H3 lysine 4 (H3K4) trimethylation at the FLC locus. This increase in trimethylation requires a PAF1-like complex and EARLY FLOWERING IN SHORT DAYS (EFS), a putative histone H3 methyltransferase. We have identified a putative zinc-finger-containing transcription factor, SUF4, that is required for the upregulation of FLC by FRI. suf4 mutations strongly suppress the late flowering phenotype of FRI, but only weakly suppress AP mutants. As with mutants in efs or the PAF1-like complex, suf4 mutants show reduced H3K4 trimethylation at FLC. An interesting distinction between the phenotypes of suf4 mutants and mutants in efs or the PAF1-like complex is observed in the expression of genes that are adjacent to FLC or FLC-like genes. In efs and PAF1-like-complex mutants, the expression of FLC, FLC-like genes and adjacent genes is suppressed. In suf4 mutants, however, only FLC expression is suppressed. These data are consistent with a model in which SUF4 may act to specifically recruit EFS and the PAF1-like complex to the FLC locus. PMID- 17079265 TI - Arabidopsis HAP2 (GCS1) is a sperm-specific gene required for pollen tube guidance and fertilization. AB - In flowering plants, sperm cells develop in the pollen cytoplasm and are transported through floral tissues to an ovule by a pollen tube, a highly polarized cellular extension. After targeting an ovule, the pollen tube bursts, releasing two sperm that fertilize an egg and a central cell. Here, we identified the gene encoding Arabidopsis HAP2, demonstrating that it is allelic to GCS1. HAP2 is expressed only in the haploid sperm and is required for efficient pollen tube guidance to ovules. We identified an insertion (hap2-1) that disrupts the C terminal portion of the protein and tags mutant pollen grains with the beta glucuronidase reporter. By monitoring reporter expression, we showed that hap2-1 does not diminish pollen tube length in vitro or in the pistil, but it reduces ovule targeting by twofold. In addition, we show that the hap2 sperm that are delivered to ovules fail to initiate fertilization. HAP2 is predicted to encode a protein with an N-terminal secretion signal, a single transmembrane domain and a C-terminal histidine-rich domain. These results point to a dual role for HAP2, functioning in both pollen tube guidance and in fertilization. Moreover, our findings suggest that sperm, long considered to be passive cargo, are involved in directing the pollen tube to its target. PMID- 17079266 TI - A dynamic fate map of the forebrain shows how vertebrate eyes form and explains two causes of cyclopia. AB - Mechanisms for shaping and folding sheets of cells during development are poorly understood. An example is the complex reorganisation of the forebrain neural plate during neurulation, which must fold a sheet into a tube while evaginating two eyes from a single contiguous domain within the neural plate. We, for the first time, track these cell rearrangements to show that forebrain morphogenesis differs significantly from prior hypotheses. We postulate a new model for forebrain neurulation and demonstrate how mutations affecting two signalling pathways can generate cyclopic phenotypes by disrupting normal cell movements or introducing new erroneous behaviours. PMID- 17079267 TI - Plant development: new models and approaches bring progress. PMID- 17079268 TI - JAK/STAT signaling promotes regional specification by negatively regulating wingless expression in Drosophila. AB - During development, a small number of conserved signaling molecules regulate regional specification, in which uniform populations of cells acquire differences and ultimately give rise to distinct organs. In the Drosophila eye imaginal disc, Wingless (Wg) signaling defines the region that gives rise to head tissue. JAK/STAT signaling was thought to regulate growth of the eye disc but not pattern formation. However, we show that the JAK/STAT pathway plays an important role in patterning the eye disc: it promotes formation of the eye field through repression of the wg gene. Overexpression of the JAK/STAT activating ligand Unpaired in the eye leads to loss of wg expression and ectopic morphogenetic furrow initiation from the lateral margins. Conversely, tissue lacking stat92E, which cannot transduce JAK/STAT signals, is transformed from retinal tissue into head cuticle, a phenotype that is also observed with ectopic Wg signaling. Consistent with this, cells lacking stat92E exhibit ectopic wg expression. Conversely, wg is autonomously repressed in cells with hyperactivated Stat92E. Furthermore, we show that the JAK/STAT pathway regulates a small enhancer in the wg 3' cis genomic region. As this enhancer is devoid of Stat92E-binding elements, we conclude that Stat92E represses wg through another, as yet unidentified factor that is probably a direct target of Stat92E. Taken together, our study is the first to demonstrate a role for the JAK/STAT pathway in regional specification by acting antagonistically to wg. PMID- 17079269 TI - Frizzled3a and Celsr2 function in the neuroepithelium to regulate migration of facial motor neurons in the developing zebrafish hindbrain. AB - Migration of neurons from their birthplace to their final target area is a crucial step in brain development. Here, we show that expression of the off limits/frizzled3a (olt/fz3a) and off-road/celsr2 (ord/celsr2) genes in neuroepithelial cells maintains the facial (nVII) motor neurons near the pial surface during their caudal migration in the zebrafish hindbrain. In the absence of olt/fz3a expression in the neuroepithelium, nVII motor neurons extended aberrant radial processes towards the ventricular surface and mismigrated radially to the dorsomedial part of the hindbrain. Our findings reveal a novel role for these genes, distinctive from their already known functions, in the regulation of the planar cell polarity (i.e. preventing integration of differentiated neurons into the neuroepithelial layer). This contrasts markedly with their reported role in reintegration of neuroepithelial daughter cells into the neuroepithelial layer after cell division. PMID- 17079270 TI - Cdx-Hox code controls competence for responding to Fgfs and retinoic acid in zebrafish neural tissue. AB - Fibroblast growth factor (Fgf) and retinoic acid (RA) signals control the formation and anteroposterior patterning of posterior hindbrain. They are also involved in development processes in other regions of the embryo. Therefore, responsiveness to Fgf and RA signals must be controlled in a context-dependent manner. Inhibiting the caudal-related genes cdx1a and cdx4 in zebrafish embryos caused ectopic expression of genes that are normally expressed in the posterior hindbrain and anterior spinal cord, and ectopic formation of the hindbrain motor and commissure neurons in the posteriormost neural tissue. Combinational marker analyses suggest mirror-image duplication in the Cdx1a/4-defective embryos, and cell transplantation analysis further revealed that Cdx1a and Cdx4 repress a posterior hindbrain-specific gene expression cell-autonomously in the posterior neural tissue. Expression of fgfs and retinaldehyde dehydrogenase 2 suggested that in the Cdx1a/4-defective embryos, the Fgf and RA signaling activities overlap in the posterior body and display opposing gradients, compared with those in the hindbrain region. We found that Fgf and RA signals were required for ectopic expression. Expression of the posterior hox genes hoxb7a, hoxa9a or hoxb9a, which function downstream of Cdx1a/4, or activator fusion genes of hoxa9a or hoxb9a (VP16-hoxa9a, VP16-hoxb9a) suppressed this loss-of-function phenotype. These data suggest that Cdx suppresses the posterior hindbrain fate through regulation of the posterior hox genes; the posterior Hox proteins function as transcriptional activators and indirectly repress the ectopic expression of the posterior hindbrain genes in the posterior neural tissue. Our results indicate that the Cdx-Hox code modifies tissue competence to respond to Fgfs and RA in neural tissue. PMID- 17079271 TI - Cytoplasmic activated protein kinase Akt regulates lipid-droplet accumulation in Drosophila nurse cells. AB - The insulin/insulin-like growth factor signalling (IIS) cascade performs a broad range of evolutionarily conserved functions, including the regulation of growth, developmental timing and lifespan, and the control of sugar, protein and lipid metabolism. Recently, these functions have been genetically dissected in the fruit fly Drosophila melanogaster, revealing a crucial role for cell-surface activation of the downstream effector kinase Akt in many of these processes. However, the mechanisms regulating lipid metabolism and the storage of lipid during development are less well characterized. Here, we use the nutrient-storing nurse cells of the fly ovary to study the cellular effects of intracellular IIS components on lipid accumulation. These cells normally store lipid in a perinuclear pool of small neutral triglyceride-containing droplets. We find that loss of the IIS signalling antagonist PTEN, which stimulates cell growth in most developing tissues, produces a very different phenotype in nurse cells, inducing formation of highly enlarged lipid droplets. Furthermore, we show that the accumulation of activated Akt in the cytoplasm is responsible for this phenotype and leads to a much higher expression of LSD2, the fly homologue of the vertebrate lipid-storage protein perilipin. Our work therefore reveals a signalling mechanism by which the effect of insulin on lipid metabolism could be regulated independently of some of its other functions during development and adulthood. We speculate that this mechanism could be important in explaining the well-established link between obesity and insulin resistance that is observed in Type 2 diabetes. PMID- 17079272 TI - Grainyhead-related transcription factor is required for duct maturation in the salivary gland and the kidney of the mouse. AB - Duct epithelial structure is an essential feature of many internal organs, including exocrine glands and the kidney. The ducts not only mediate fluid transfer but also help to maintain homeostasis. For instance, fluids and solutes are resorbed from or secreted into the primary fluid flowing through the lumen of the ducts in the exocrine glands and kidneys. The molecular mechanism underlying the functional maturation of these ducts remains largely unknown. Here, we show that a grainyhead-related transcription factor, CP2-like 1 (CP2L1), is required for the maturation of the ducts of the salivary gland and kidney. In the mouse, Cp2l1 is specifically expressed in the developing ducts of a number of exocrine glands, including the salivary gland, as well as in those of the kidney. In Cp2l1 deficient mice, the expression of genes directly involved in functional maturation of the ducts was specifically reduced in both the salivary gland and kidney, indicating that Cp2l1 is required for the differentiation of duct cells. Furthermore, the composition of saliva and urine was abnormal in these mice. These results indicate that Cp2l1 expression is required for normal duct development in both the salivary gland and kidney. PMID- 17079273 TI - POL and PLL1 phosphatases are CLAVATA1 signaling intermediates required for Arabidopsis shoot and floral stem cells. AB - The post-embryonic development of above-ground tissues in plants is dependent upon the maintenance and differentiation of stem cells at the shoot meristem. The Arabidopsis WUSCHEL (WUS) transcription factor establishes an organizing center within the shoot meristem that is essential for specification of stem-cell identity in overlying cells. The CLAVATA (CLV) signaling pathway, including the CLV1 receptor-kinase, promotes the differentiation of stem cells by limiting the WUS expression domain, yet the mechanism of CLV signaling is largely unknown. Previously, we have shown that mutations in two protein phosphatases, POLTERGEIST (POL) and PLL1, partially suppress clv mutant phenotypes. Here, we demonstrate that POL and PLL1 are integral components of the CLV1 signaling pathway. POL and PLL1 are essential for stem-cell specification, and can also block stem-cell differentiation when overexpressed. We provide extensive evidence that POL and PLL1 act downstream of CLV signaling to maintain WUS expression and that they regulate WUS at a transcriptional level. Our findings suggest that POL and PLL1 are central players in regulating the balance between stem-cell maintenance and differentiation, and are the closest known factors to WUS regulation in the shoot meristem. PMID- 17079274 TI - Bchs, a BEACH domain protein, antagonizes Rab11 in synapse morphogenesis and other developmental events. AB - BEACH proteins, an evolutionarily conserved family characterized by the presence of a BEACH (Beige and Chediak-Higashi) domain, have been implicated in membrane trafficking, but how they interact with the membrane trafficking machinery is unknown. Here we show that the Drosophila BEACH protein Bchs (Blue cheese) acts during development as an antagonist of Rab11, a small GTPase involved in vesicle trafficking. We find that reduction in, or loss of, bchs function restores viability and normal bristle development in animals with reduced rab11 function, while reductions in rab11 function exacerbate defects caused by bchs overexpression in the eye. Consistent with a role for Bchs in modulating Rab11 dependent trafficking, Bchs protein is associated with vesicles and extensively colocalized with Rab11 at the neuromuscular junction (NMJ). At the NMJ, we find that rab11 is important for synaptic morphogenesis, as reductions in rab11 function cause increases in bouton density and branching. These defects are also suppressed by loss of bchs. Taken together, these data identify Bchs as an antagonist of Rab11 during development and uncover a role for these regulators of vesicle trafficking in synaptic morphogenesis. This raises the interesting possibility that Bchs and other BEACH proteins may regulate vesicle traffic via interactions with Rab GTPases. PMID- 17079276 TI - Visibility of Si nanoparticles embedded in an amorphous SiO2 matrix. AB - The visibility of a Si crystalline nanoparticle of diameter 2 nm embedded in an amorphous SiO(2) layer is evaluated quantitatively by multislice calculation. The visibility depends on the crystal orientation of the Si nanoparticle, the thickness of the amorphous SiO(2) layer and the defocus. Scherzer defocus always gives the highest visibility at any crystal orientation. The visibility is higher when the incident beam is parallel to the (111) planes and the (111) fringes are most visible. The image of a Si nanoparticle is obscured by random images from the amorphous SiO(2) layer and the Si nanoparticle becomes invisible when it is misoriented or the amorphous layer is thicker than 60 nm. The probability that a Si nanoparticle can be distinguished from the random noise of amorphous images is 89% when the thickness of the amorphous SiO(2) layer is 12 nm, but this is reduced to 21% when the layer is 48 nm thick. These quantitative results are useful when estimating the density of Si nanoparticles including invisible nanoparticles. PMID- 17079275 TI - DeltaNp63 plays an anti-apoptotic role in ventral bladder development. AB - The bladder, the largest smooth-muscle organ in the human body, is responsible for urine storage and micturition. P63, a homolog of the p53 tumor-suppressor gene, is essential for the development of all stratified epithelia, including the bladder urothelium. The N-terminal truncated isoform of p63, DeltaNp63, is known to have anti-apoptotic characteristics. We have established that DeltaNp63 is not only the predominant isoform expressed throughout the bladder, but is also preferentially expressed in the ventral bladder urothelium during early development. We observed a host of ventral defects in p63-/- embryos, including the absence of the abdominal and ventral bladder walls. This number of ventral defects is identical to bladder exstrophy, a congenital anomaly exhibited in human neonates. In the absence of p63, the ventral urothelium was neither committed nor differentiated, whereas the dorsal urothelium was both committed and differentiated. Furthermore, in p63-/- bladders, apoptosis in the ventral urothelium was significantly increased. This was accompanied by the upregulation of mitochondrial apoptotic mediators Bax and Apaf1, and concurrent upregulation of p53. Overexpression of DeltaNp63gamma and DeltaNp63beta in p63-/- bladder primary cell cultures resulted in a rescue, evidenced by significantly reduced expressions of Bax and Apaf1. We conclude that DeltaNp63 plays a crucial anti apoptotic role in normal bladder development. PMID- 17079278 TI - The natural radioactivity in the bricks used for the construction of the dwelling in Tehran areas of Iran. AB - This paper presents the finding of a study undertaken to determine the natural radioactivity present in bricks in Tehran. A total of 45 samples of commonly used bricks were collected from various manufacturers and suppliers and analyzed using a shielded HPGe gamma ray spectrometer. The specific activities of (226) Ra, (232) Th, and (40) K were measured in all brick samples, and results ranged from minimum values of 26, 24, and 524 Bq kg(-1) to maximum values of 38, 36, and 820 Bq kg(-1) with mean values of 33, 30, and 700 Bq kg(-1) ,respectively. In general, these activities were comparable to the results of similar studies undertaken in another countries. These activities would suggest that the use of such bricks in construction of dwellings or work places in Tehran is unlikely to give rise to any significant radiation exposure to the occupants. PMID- 17079279 TI - Hsp70 negatively controls rotavirus protein bioavailability in caco-2 cells infected by the rotavirus RF strain. AB - Previous studies demonstrated that the induction of the heat shock protein Hsp70 in response to viral infection is highly specific and differs from one cell to another and for a given virus type. However, no clear consensus exists so far to explain the likely reasons for Hsp70 induction within host cells during viral infection. We show here that upon rotavirus infection of intestinal cells, Hsp70 is indeed rapidly, specifically, and transiently induced. Using small interfering RNA-Hsp70-transfected Caco-2 cells, we observed that Hsp70 silencing was associated with an increased virus protein level and enhanced progeny virus production. Upon Hsp70 silencing, we observed that the ubiquitination of the main rotavirus structural proteins was strongly reduced. In addition, the use of proteasome inhibitors in infected Caco-2 cells was shown to induce an accumulation of structural viral proteins. Together, these results are consistent with a role of Hsp70 in the control of the bioavailability of viral proteins within cells for virus morphogenesis. PMID- 17079280 TI - Control of simian immunodeficiency virus SIVmac239 is not predicted by inheritance of Mamu-B*17-containing haplotypes. AB - It is well established that host genetics, especially major histocompatibility complex (MHC) genes, are important determinants of human immunodeficiency virus disease progression. Studies with simian immunodeficiency virus (SIV)-infected Indian rhesus macaques have associated Mamu-B*17 with control of virus replication. Using microsatellite haplotyping of the 5-Mb MHC region, we compared disease progression among SIVmac239-infected Indian rhesus macaques that possess Mamu-B*17-containing MHC haplotypes that are identical by descent. We discovered that SIV-infected animals possessing identical Mamu-B*17-containing haplotypes had widely divergent disease courses. Our results demonstrate that the inheritance of a particular Mamu-B*17-containing haplotype is not sufficient to predict SIV disease outcome. PMID- 17079281 TI - The level of CD81 cell surface expression is a key determinant for productive entry of hepatitis C virus into host cells. AB - Recently a cell culture model supporting the complete life cycle of the hepatitis C virus (HCV) was developed. Searching for host cell determinants involved in the HCV replication cycle, we evaluated the efficiency of virus propagation in different Huh-7-derived cell clones. We found that Huh-7.5 cells and Huh7-Lunet cells, two former replicon cell clones that had been generated by removal of an HCV replicon by inhibitor treatment, supported comparable levels of RNA replication and particle production, whereas virus spread was severely impaired in the latter cells. Analysis of cell surface expression of CD81 and scavenger receptor class B type I (SR-BI), two molecules previously implicated in HCV entry, revealed similar expression levels for SR-BI, while CD81 surface expression was much higher on Huh-7.5 cells than on Huh7-Lunet cells. Ectopic expression of CD81 in Huh7-Lunet cells conferred permissiveness for HCV infection to a level comparable to that for Huh-7.5 cells. Modulation of CD81 cell surface density in Huh-7.5 cells by RNA interference indicated that a certain amount of this molecule (approximately 7 x 10(4) molecules per cell) is required for productive infection with a low dose of HCV. Consistent with this, we show that susceptibility to HCV infection depends on a critical quantity of CD81 molecules. While infection is restricted in cells expressing very small amounts of CD81, susceptibility rapidly rises within a narrow range of CD81 levels, reaching a plateau where higher expression does not further increase the efficiency of infection. Together these data indicate that a high density of cell surface exposed CD81 is a key determinant for productive HCV entry into host cells. PMID- 17079282 TI - Compensatory mutations in E1, p7, NS2, and NS3 enhance yields of cell culture infectious intergenotypic chimeric hepatitis C virus. AB - There is little understanding of mechanisms underlying the assembly and release of infectious hepatitis C virus (HCV) from cultured cells. Cells transfected with synthetic genomic RNA from a unique genotype 2a virus (JFH1) produce high titers of virus, while virus yields are much lower with a prototype genotype 1a RNA containing multiple cell culture-adaptive mutations (H77S). To characterize the basis for this difference in infectious particle production, we constructed chimeric genomes encoding the structural proteins of H77S within the background of JFH1. RNAs encoding polyproteins fused at the NS2/NS3 junction ("H-NS2/NS3-J") and at a site of natural, intergenotypic recombination within NS2 ["H-(NS2)-J"] produced infectious virus. In contrast, no virus was produced by a chimera fused at the p7-NS2 junction. Chimera H-NS2/NS3-J virus (vH-NS2/NS3-J) recovered from transfected cultures contained compensatory mutations in E1 and NS3 that were essential for the production of infectious virus, while yields of infectious vH (NS2)-J were enhanced by mutations within p7 and NS2. These compensatory mutations were chimera specific and did not enhance viral RNA replication or polyprotein processing; thus, they likely compensate for incompatibilities between proteins of different genotypes at sites of interactions essential for virus assembly and/or release. Mutations in p7 and NS2 acted additively and increased the specific infectivity of vH-(NS2)-J particles, while having less impact on the numbers of particles released. We conclude that interactions between NS2 and E1 and p7 as well as between NS2 and NS3 are essential for virus assembly and/or release and that each of these viral proteins plays an important role in this process. PMID- 17079283 TI - Coordinated regulation and widespread cellular expression of interferon stimulated genes (ISG) ISG-49, ISG-54, and ISG-56 in the central nervous system after infection with distinct viruses. AB - The interferon (IFN)-stimulated genes (ISGs) ISG-49, ISG-54, and ISG-56 are highly responsive to viral infection, yet the regulation and function of these genes in vivo are unknown. We examined the simultaneous regulation of these ISGs in the brains of mice during infection with either lymphocytic choriomeningitis virus (LCMV) or West Nile virus (WNV). Expression of the ISG-49 and ISG-56 genes increased significantly during LCMV infection, being widespread and localized predominantly to common as well as distinct neuronal populations. Expression of the ISG-54 gene also increased but to lower levels and with a more restricted distribution. Although expression of the ISG-49, ISG-54, and ISG-56 genes was increased in the brains of LCMV-infected STAT1 and STAT2 knockout (KO) mice, this was blunted, delayed, and restricted to the choroid plexus, meninges, and endothelium. ISG-56 protein was regulated in parallel with the corresponding RNA transcript in the brain during LCMV infection in wild-type and STAT KO mice. Similar changes in ISG-49, ISG-54, and ISG-56 RNA levels and ISG-56 protein levels were observed in the brains of wild-type mice following infection with WNV. Thus, the ISG-49, ISG-54, and ISG-56 genes are coordinately upregulated in the brain during LCMV and WNV infection; this upregulation, in the case of LCMV, was totally (neurons) or partially (non-neurons) dependent on the IFN-signaling molecules STAT1 and STAT2. These findings suggest a dominant role for the ISG-49, ISG-54, and ISG-56 genes in the host response to different viruses in the central nervous system, where, particularly in neurons, these genes may have nonredundant functions. PMID- 17079284 TI - Tubulovesicular structures within vesicular stomatitis virus G protein pseudotyped lentiviral vector preparations carry DNA and stimulate antiviral responses via Toll-like receptor 9. AB - Recombinant lentiviral vectors (LVs) are commonly used as research tools and are being tested in the clinic as delivery agents for gene therapy. Here, we show that Vesicular stomatitis virus G protein (VSV-G)-pseudotyped LV preparations produced by transient transfection are heavily contaminated with tubulovesicular structures (TVS) of cellular origin, which carry nucleic acids, including the DNA plasmids originally used for LV generation. The DNA carried by TVS can act as a stimulus for innate antiviral responses, triggering Toll-like receptor 9 and inducing alpha/beta interferon production by plasmacytoid dendritic cells (pDC). Removal of TVS markedly reduces the ability of VSV-G-pseudotyped LV preparations to activate pDC. Conversely, virus-free TVS are sufficient to stimulate pDC and act as potent adjuvants in vivo, eliciting T- and B-cell responses to coadministered proteins. These results highlight the role of by-products of virus production in determining the immunostimulatory properties of recombinant virus preparations and suggest possible strategies for diminishing responses to LVs in gene therapy and in research use. PMID- 17079285 TI - Genetic protection against hepatitis B virus conferred by CCR5Delta32: Evidence that CCR5 contributes to viral persistence. AB - Recovery from acute hepatitis B virus (HBV) infection requires a broad, vigorous T-cell response, which is enhanced in mice when chemokine receptor 5 (CCR5) is missing. To test the hypothesis that production of a nonfunctional CCR5 (CCR5Delta32 [a functionally null allele containing a 32-bp deletion]) increases the likelihood of recovery from hepatitis B in humans, we studied 526 persons from three cohorts in which one person with HBV persistence was matched to two persons who recovered from an HBV infection. Recovery or persistence was determined prior to availability of lamivudine. We determined genotypes for CCR5Delta32 and for polymorphisms in the CCR5 promoter and in coding regions of the neighboring genes, chemokine receptor 2 (CCR2) and chemokine receptor-like 2 (CCRL2). Allele and haplotype frequencies were compared among the 190 persons with viral recovery and the 336 with persistence by use of conditional logistic regression. CCR5Delta32 reduced the risk of developing a persistent HBV infection by nearly half (odds ratio [OR], 0.53; 95% confidence interval [CI], 0.33 to 0.83; P = 0.006). This association was virtually identical in persons with and without a concomitant human immunodeficiency virus infection. Of the nine individuals who were homozygous for the deletion, eight recovered from infection (OR, 0.25; 95% CI, 0.03 to 1.99; P = 0.19). None of the other neighboring polymorphisms examined were associated with HBV outcome. These data demonstrate a protective effect of CCR5Delta32 in recovery from an HBV infection, provide genetic epidemiological evidence for a role of CCR5 in the immune response to HBV, and suggest a potential therapeutic treatment for patients persistently infected with HBV. PMID- 17079286 TI - Double-stranded RNA deaminase ADAR1 increases host susceptibility to virus infection. AB - The RNA-editing enzyme ADAR1 is a double-stranded RNA (dsRNA) binding protein that modifies cellular and viral RNA sequences by adenosine deamination. ADAR1 has been demonstrated to play important roles in embryonic erythropoiesis, viral response, and RNA interference. In human hepatitis virus infection, ADAR1 has been shown to target viral RNA and to suppress viral replication through dsRNA editing. It is not clear whether this antiviral effect of ADAR1 is a common mechanism in response to viral infection. Here, we report a proviral effect of ADAR1 that enhances replication of vesicular stomatitis virus (VSV) through a mechanism independent of dsRNA editing. We demonstrate that ADAR1 interacts with dsRNA-activated protein kinase PKR, inhibits its kinase activity, and suppresses the alpha subunit of eukaryotic initiation factor 2 (eIF-2alpha) phosphorylation. Consistent with the inhibitory effect on PKR activation, ADAR1 increases VSV infection in PKR+/+ mouse embryonic fibroblasts; however, no significant effect was found in PKR-/- cells. This proviral effect of ADAR1 requires the N-terminal domains but does not require the deaminase domain. These findings reveal a novel mechanism of ADAR1 that increases host susceptibility to viral infection by inhibiting PKR activation. PMID- 17079287 TI - Epstein-Barr virus BZLF1 gene, a switch from latency to lytic infection, is expressed as an immediate-early gene after primary infection of B lymphocytes. AB - We demonstrate here that the Epstein-Barr virus (EBV) BZLF1 gene, a switch from latent infection to lytic infection, is expressed as early as 1.5 h after EBV infection in Burkitt's lymphoma-derived, EBV-negative Akata and Daudi cells and primary B lymphocytes. Since BZLF1 mRNA is expressed even when the cells are infected with EBV in the presence of anisomycin, an inhibitor of protein synthesis, its expression does not require prerequisite protein synthesis, indicating that BZLF1 is expressed as an immediate-early gene following primary EBV infection of B lymphocytes. PMID- 17079288 TI - Analysis of CD127 and KLRG1 expression on hepatitis C virus-specific CD8+ T cells reveals the existence of different memory T-cell subsets in the peripheral blood and liver. AB - The differentiation and functional status of virus-specific CD8+ T cells is significantly influenced by specific and ongoing antigen recognition. Importantly, the expression profiles of the interleukin-7 receptor alpha chain (CD127) and the killer cell lectin-like receptor G1 (KLRG1) have been shown to be differentially influenced by repetitive T-cell receptor interactions. Indeed, antigen-specific CD8+ T cells targeting persistent viruses (e.g., human immunodeficiency virus and Epstein-Barr virus) have been shown to have low CD127 and high KLRG1 expressions, while CD8+ T cells targeting resolved viral antigens (e.g., FLU) typically display high CD127 and low KLRG1 expressions. Here, we analyzed the surface phenotype and function of hepatitis C virus (HCV)-specific CD8+ T cells. Surprisingly, despite viral persistence, we found that a large fraction of peripheral HCV-specific CD8+ T cells were CD127+ and KLRG1- and had good proliferative capacities, thus resembling memory cells that usually develop following acute resolving infection. Intrahepatic virus-specific CD8+ T cells displayed significantly reduced levels of CD127 expression but similar levels of KLRG1 expression compared to the peripheral blood. These results extend previous studies that demonstrated central memory (CCR7+) and early-differentiated phenotypes of HCV-specific CD8+ T cells and suggest that insufficient stimulation of virus-specific CD8+ T cells by viral antigen may be responsible for this alteration in HCV-specific CD8+ T-cell differentiation during chronic HCV infection. PMID- 17079289 TI - Inhibition of retinoic acid-inducible gene I-mediated induction of beta interferon by the NS1 protein of influenza A virus. AB - The retinoic acid-inducible gene I product (RIG-I) has been identified as a cellular sensor of RNA virus infection resulting in beta interferon (IFN-beta) induction. However, many viruses are known to encode viral products that inhibit IFN-beta production. In the case of influenza A virus, the viral nonstructural protein 1 (NS1) prevents the induction of the IFN-beta promoter by inhibiting the activation of transcription factors, including IRF-3, involved in IFN-beta transcriptional activation. The inhibitory properties of NS1 appear to be due at least in part to its binding to double-stranded RNA (dsRNA), resulting in the sequestration of this viral mediator of RIG-I activation. However, the precise effects of NS1 on the RIG-I-mediated induction of IFN-beta have not been characterized. We now report that the NS1 of influenza A virus interacts with RIG I and inhibits the RIG-I-mediated induction of IFN-beta. This inhibition was apparent even when a mutant RIG-I that is constitutively activated (in the absence of dsRNA) was used to trigger IFN-beta production. Coexpression of RIG-I, its downstream signaling partner, IPS-1, and NS1 resulted in increased levels of RIG-I and NS1 within an IPS-1-rich, solubilization-resistant fraction after cell lysis. These results suggest that RIG-I, IPS-1, and NS1 become part of the same complex. Consistent with this idea, NS1 was also found to inhibit IFN-beta promoter activation by IPS-1 overexpression. Our results indicate that, in addition to sequestering dsRNA, the NS1 of influenza A virus binds to RIG-I and inhibits downstream activation of IRF-3, preventing the transcriptional induction of IFN-beta. PMID- 17079290 TI - Efficient incorporation of tegument proteins pUL46, pUL49, and pUS3 into pseudorabies virus particles depends on the presence of pUL21. AB - The mature virion of the alphaherpesvirus pseudorabies virus (PrV) contains a minimum of 31 structural proteins which are recruited into the virus particle by a network of protein-protein interactions which is only incompletely understood. We show here that deletion of the tegument protein pUL21 resulted in a drastic decrease in the incorporation of the pUL46, pUL49, and pUS3 tegument components into mature virions. Moreover, the attenuated PrV strain Bartha (PrV-Ba), which, among other defects, carries mutations in pUL21, also fails to package pUL46, pUL49, and pUS3 efficiently. By the reconstitution of wild-type pUL21 expression to PrV-Ba and the transfer of mutated PrV-Ba pUL21 into wild-type PrV, we demonstrate that this phenotype is due to the mutated pUL21. PMID- 17079291 TI - Varicella-zoster virus infection of human fibroblast cells activates the c-Jun N terminal kinase pathway. AB - The transcription factors ATF-2 and c-Jun are important for transactivation of varicella-zoster virus (VZV) genes. c-Jun is activated by the c-Jun N-terminal kinase (JNK), a member of the mitogen-activated protein kinase pathway that responds to stress and cytokines. To study the effects of VZV on this pathway, confluent human foreskin fibroblasts were infected with cell-associated VZV for 1 to 4 days. Immunoblots showed that phosphorylated JNK and c-Jun levels increased in VZV-infected cells, and kinase assays determined that phospho-JNK was active. Phospho-JNK was detected after 24 h, and levels rose steadily over 4 days in parallel with accumulation of VZV antigen. The two main activators of JNK are MKK4 and MKK7, and levels of their active, phosphorylated forms also increased. The competitive inhibitor of JNK, SP600125, caused a dose-dependent reduction in VZV yield (50% effective concentration, congruent with 8 microM). Specificity was verified by immunoblotting; phospho-c-Jun was eliminated by 18 microM SP600125 in VZV-infected cells. Immunofluorescent confocal microscopy showed that phospho-c Jun and most of phospho-JNK were in the nuclei of VZV-infected cells; some phospho-JNK was in the cytoplasm. MKK4, MKK7, JNK, and phospho-JNK were detected by immunoblotting in purified preparations of VZV virions, but c-Jun was absent. JNK was located in the virion tegument, as determined by biochemical fractionation and immunogold transmission electron microscopy. Overall, these results demonstrate the importance of the JNK pathway for VZV replication and advance the idea that JNK is a useful drug target against VZV. PMID- 17079292 TI - Inefficient human immunodeficiency virus replication in mobile lymphocytes. AB - Cell-to-cell viral transfer facilitates the spread of lymphotropic retroviruses such as human immunodeficiency virus (HIV) and human T-cell leukemia virus (HTLV), likely through the formation of "virological synapses" between donor and target cells. Regarding HIV replication, the importance of cell contacts has been demonstrated, but this phenomenon remains only partly characterized. In order to alter cell-to-cell HIV transmission, we have maintained cultures under continuous gentle shaking and followed viral replication in this experimental system. In lymphoid cell lines, as well as in primary lymphocytes, viral replication was dramatically reduced in shaken cultures. To document this phenomenon, we have developed an assay to assess the relative contributions of free and cell associated virions in HIV propagation. Acutely infected donor cells were mixed with carboxyfluorescein diacetate succinimidyl ester-labeled lymphocytes as targets, and viral production was followed by measuring HIV Gag expression at different time points by flow cytometry. We report that cellular contacts drastically enhance productive viral transfer compared to what is seen with infection with free virus. Productive cell-to-cell viral transmission required fusogenic viral envelope glycoproteins on donor cells and adequate receptors on targets. Only a few syncytia were observed in this coculture system. Virus release from donor cells was unaffected when cultures were gently shaken, whereas virus transfer to recipient cells was severely impaired. Altogether, these results indicate that cell-to-cell transfer is the predominant mode of HIV spread and help to explain why this virus replicates so efficiently in lymphoid organs. PMID- 17079293 TI - Presence of a surface-exposed loop facilitates trypsinization of particles of Sinsiro virus, a genogroup II.3 norovirus. AB - Noroviruses (NoVs) are the causative agents of nonbacterial acute gastroenteritis in humans. NoVs that belong to genogroup II (GII) are quite prevalent and prone to undergo recombination, and their three-dimensional structure is not yet known. Protein homology modeling of Sinsiro virus (SV), a member of the GII.3 NoVs, revealed the presence of a surface-exposed 20-amino-acid (aa) insertion in the P2 domain of the capsid protein (CP) relative to the Norwalk virus (NV) CP, which is a well known hot spot for mutations to counter the host immunological response. To further characterize the role of the long insertion in SV, the capsid protein gene was expressed using the recombinant baculovirus system. Trypsinization of the resultant virus-like particles yielded two predominant bands (31.7 and 26.1 kDa) in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis. N-terminal sequencing and analysis of the mass spectroscopic data indicated that these fragments correspond to residues 1 to 292 (26.1 kDa) and 307 to 544 (31.7 kDa). In addition, the above data taken together with the comparative modeling studies indicated that the trypsin cleavage sites of the Sinsiro virus CP, Arg292 and Arg307, are located at the beginning of and within the 20-aa insertion in the P2 domain, respectively. This study demonstrates that the presence of the surface-exposed loop in the GII.3 NoVs facilitates the trypsinization of the capsid protein in the assembled form. The SV particles remain intact even after trypsin digestion and retain the suggested receptor binding linear epitope of residues 325 to 334. The above results are distinct from those obtained from the trypsinization studies performed earlier on the NV (GI) and VA387 (GII) viruses, both of which lack the large surface insertion and associated basic residues. These new observations may have implications for host receptor binding, cell entry, and norovirus infection in general. PMID- 17079294 TI - Immunogenic and functional organization of hepatitis C virus (HCV) glycoprotein E2 on infectious HCV virions. AB - Development of full-length hepatitis C virus (HCV) RNAs replicating efficiently and producing infectious cell-cultured virions, HCVcc, in hepatoma cells provides an opportunity to characterize immunogenic domains on viral envelope proteins involved in entry into target cells. A panel of immunoglobulin G1 human monoclonal antibodies (HMAbs) to three immunogenic conformational domains (designated A, B, and C) on HCV E2 glycoprotein showed that epitopes within two domains, B and C, mediated HCVcc neutralization, whereas HMAbs to domain A were all nonneutralizing. For the neutralizing antibodies to domain B (with some to conserved epitopes among different HCV genotypes), the inhibitory antibody concentration reducing HCVcc infection by 90%, IC90, ranged from 0.1 to 4 microg/ml. For some neutralizing HMAbs, HCVcc neutralization displayed a linear correlation with an antibody concentration between the IC50 and the IC90 while others showed a nonlinear correlation. The differences between IC50/IC90 ratios and earlier findings that neutralizing HMAbs block E2 interaction with CD81 suggest that these antibodies block different facets of virus-receptor interaction. Collectively, these findings support an immunogenic model of HCV E2 having three immunogenic domains with distinct structures and functions and provide added support for the idea that CD81 is required for virus entry. PMID- 17079295 TI - Sheep-passaged bovine spongiform encephalopathy agent exhibits altered pathobiological properties in bovine-PrP transgenic mice. AB - Sheep can be experimentally infected with bovine spongiform encephalopathy (BSE), and the ensuing disease is similar to scrapie in terms of pathogenesis and clinical signs. BSE infection in sheep is an animal and human health concern. In this study, the transmission in BoPrP-Tg110 mice of prions from BSE-infected sheep was examined and compared to the transmission of original cattle BSE in cattle and sheep scrapie prions. Our results indicate no transmission barrier for sheep BSE prions to infect BoPrP-Tg110 mice, but the course of the disease is accelerated compared to the effects of the original BSE isolate. The shortened incubation period of sheep BSE in the model was conserved in subsequent passage in BoPrP-Tg110 mice, indicating that it is not related to infectious titer differences. Biochemical signature, lesion profile, and PrP(Sc) deposition pattern of both cattle and sheep BSE were similar. In contrast, all three sheep scrapie isolates tested showed an evident transmission barrier and further adaptation in subsequent passage. Taken together, those data indicate that BSE agent can be altered by crossing a species barrier, raising concerns about the virulence of this new prion towards other species, including humans. The BoPrP Tg110 mouse bioassay should be considered as a valuable tool for discriminating scrapie and BSE in sheep. PMID- 17079296 TI - Attenuation of herpes simplex virus neurovirulence with picornavirus cis-acting genetic elements. AB - Viral pathogenesis depends on a suitable milieu in target host cells permitting viral gene expression, propagation, and spread. In many instances, viral genomes can be manipulated to select for propagation in certain tissues or cell types. This has been achieved for the neurotropic poliovirus (PV) by exchange of the internal ribosomal entry site (IRES), which is responsible for translation of the uncapped plus-strand RNA genome. The IRES of human rhinovirus type 2 (HRV2) confers neuron-specific replication deficits to PV but has no effect on viral propagation in malignant glioma cells. We report here that placing the critical gamma(1)34.5 virulence genes of herpes simplex virus type 1 (HSV) under translation control of the HRV2 IRES results in neuroattenuation in mice. In contrast, IRES insertion permits HSV propagation in malignant glioma cell lines that do not support replication of HSV recombinants carrying gamma(1)34.5 deletions. Our observations indicate that the conditions for alternative translation initiation at the HRV2 IRES in malignant glioma cells differ from those in normal central nervous system (CNS) cells. Picornavirus regulatory sequences mediating cell type-specific gene expression in the CNS can be utilized to target cancerous cells at the level of translation regulation outside their natural context. PMID- 17079297 TI - Adenovirus ubiquitin-protein ligase stimulates viral late mRNA nuclear export. AB - Theadenovirus type 5 (Ad5) E1B-55K and E4orf6 proteins are required together to stimulate viral late nuclear mRNA export to the cytoplasm and to restrict host cell nuclear mRNA export during the late phase of infection. Previous studies have shown that these two viral proteins interact with the cellular proteins elongins B and C, cullin 5, RBX1, and additional cellular proteins to form an E3 ubiquitin-protein ligase that polyubiquitinates p53 and probably one or more subunits of the MRE11-RAD50-NBS1 (MRN) complex, directing their proteasomal degradation. The MRN complex is required for cellular DNA double-strand break repair and induction of the DNA damage response by adenovirus infection. To determine if the ability of E1B-55K and E4orf6 to stimulate viral late mRNA nuclear export requires the ubiquitin-protein ligase activity of this viral ubiquitin-protein ligase complex, we designed and expressed a dominant-negative mutant form of cullin 5 in HeLa cells before infection with wild-type Ad5 or the E1B-55K null mutant dl1520. The dominant-negative cullin 5 protein stabilized p53 and the MRN complex, indicating that it inhibited the viral ubiquitin-protein ligase but had no effect on viral early mRNA synthesis, early protein synthesis, or viral DNA replication. However, expression of the dominant-negative cullin 5 protein caused a decrease in viral late protein synthesis and viral nuclear mRNA export similar to the phenotype produced by mutations in E1B-55K. We conclude that the stimulation of adenovirus late mRNA nuclear export by E1B-55K and E4orf6 results from the ubiquitin-protein ligase activity of the adenovirus ubiquitin protein ligase complex. PMID- 17079298 TI - The switch from latent to productive infection in epstein-barr virus-infected B cells is associated with sensitization to NK cell killing. AB - Following activation of Epstein-Barr virus (EBV)-infected B cells from latent to productive (lytic) infection, there is a concomitant reduction in the level of cell surface major histocompatibility complex (MHC) class I molecules and an impaired antigen-presenting function that may facilitate evasion from EBV specific CD8+ cytotoxic T cells. In some other herpesviruses studied, most notably human cytomegalovirus (HCMV), evasion of virus-specific CD8+ effector responses via downregulation of surface MHC class I molecules is supplemented with specific mechanisms for evading NK cells. We now report that EBV differs from HCMV in this respect. While latently infected EBV-positive B cells were resistant to lysis by two NK lines and by primary polyclonal NK cells from peripheral blood, these effectors efficiently killed cells activated into the lytic cycle. Susceptibility to NK lysis coincided not only with downregulation of HLA-A, -B, and -C molecules that bind to the KIR family of inhibitory receptors on NK cells but also with downregulation of HLA-E molecules binding the CD94/NKG2A inhibitory receptors. Conversely, ULBP-1 and CD112, ligands for the NK cell-activating receptors NKG2D and DNAM-1, respectively, were elevated. Susceptibility of the virus-producing target cells to NK cell lysis was partially reversed by blocking ULBP-1 or CD112 with specific antibodies. These results highlight a fundamental difference between EBV and HCMV with regards to evasion of innate immunity. PMID- 17079299 TI - Increased neutralization sensitivity of recently emerged CXCR4-using human immunodeficiency virus type 1 strains compared to coexisting CCR5-using variants from the same patient. AB - CXCR4-using (X4) human immunodeficiency virus type 1 (HIV-1) variants evolve from CCR5-using (R5) variants relatively late in the natural course of infection in 50% of HIV-1 subtype B-infected individuals and subsequently coexist with R5 HIV 1 variants. This relatively late appearance of X4 HIV-1 variants is poorly understood. Here we tested the neutralization sensitivity for soluble CD4 (sCD4) and the broadly neutralizing antibodies IgG1b12, 2F5, 4E10, and 2G12 of multiple coexisting clonal R5 and (R5)X4 (combined term for monotropic X4 and dualtropic R5X4 viruses) HIV-1 variants that were obtained at two time points after the first appearance of X4 variants in five participants of the Amsterdam Cohort Studies on HIV-1 infection and AIDS. Recently emerged (R5)X4 viruses were significantly more sensitive to neutralization by the CD4-binding-site-directed agents sCD4 and IgG1b12 than their coexisting R5 viruses. This difference was less pronounced at the later time point. Early (R5)X4 variants from two out of four patients were also highly sensitive to neutralization by autologous serum (50% inhibition at serum dilutions of >200). Late (R5)X4 viruses from these two patients were neutralized at lower serum dilutions, which suggested escape of X4 variants from humoral immunity. Autologous neutralization of coexisting R5 and (R5)X4 variants was not observed in the other patients. In conclusion, the increased neutralization sensitivity of HIV-1 variants during the transition from CCR5 usage to CXCR4 usage may imply an inhibitory role for humoral immunity in HIV-1 phenotype evolution in some patients, thus potentially contributing to the late emergence of X4 variants. PMID- 17079300 TI - Expression of viral microRNAs in Epstein-Barr virus-associated gastric carcinoma. AB - Epstein-Barr virus (EBV) is associated with about 6 to 16% of gastric carcinoma cases worldwide. Expression of the EBV microRNAs (miRNAs) was observed in B cells and nasopharyngeal carcinoma cells infected with EBV. However, it is not clear if the EBV miRNAs are expressed in EBV-associated gastric carcinomas (EBVaGCs). We found that BART miRNAs but not BHRF1 miRNAs were expressed in EBV-infected gastric carcinoma cell lines and the tumor tissues from patients as well as the animal model. The expression of viral miRNAs in EBVaGCs suggests that these EBV miRNAs may play important roles in the tumorigenesis of EBVaGCs. PMID- 17079301 TI - Assembly of two independent populations of flock house virus particles with distinct RNA packaging characteristics in the same cell. AB - Flock House virus (FHV; Nodaviridae) is a positive-strand RNA virus that encapsidates a bipartite genome consisting of RNA1 and RNA2. We recently showed that specific recognition of these RNAs for packaging into progeny particles requires coat protein translated from replicating viral RNA. In the present study, we investigated whether the entire assembly pathway, i.e., the formation of the initial nucleating complex and the subsequent completion of the capsid, is restricted to the same pool of coat protein subunits. To test this, coat proteins carrying either FLAG or hemagglutinin epitopes were synthesized from replicating or nonreplicating RNA in the same cell, and the resulting particle population and its RNA packaging phenotype were analyzed. Results from immunoprecipitation analysis and ion-exchange chromatography showed that the differentially tagged proteins segregated into two distinct populations of virus particles with distinct RNA packaging phenotypes. Particles assembled from coat protein that was translated from replicating RNA contained the FHV genome, whereas particles assembled from coat protein that was translated from nonreplicating mRNA contained random cellular RNA. These data demonstrate that only coat proteins synthesized from replicating RNA partake in the assembly of virions that package the viral genome and that RNA replication, coat protein translation, and virion assembly are processes that are tightly coupled during the life cycle of FHV. PMID- 17079302 TI - Adenoviruses use lactoferrin as a bridge for CAR-independent binding to and infection of epithelial cells. AB - Most adenoviruses bind to the coxsackie- and adenovirus receptor (CAR). Surprisingly, CAR is not expressed apically on polarized cells and is thus not easily available to viruses. Consequently, alternative mechanisms for entry of coxsackievirus and adenovirus into cells have been suggested. We have found that tear fluid promotes adenovirus infection, and we have identified human lactoferrin (HLf) as the tear fluid component responsible for this effect. HLf alone was found to promote binding of adenovirus to epithelial cells in a dose dependent manner and also infection of epithelial cells by adenovirus. HLf was also found to promote gene delivery from an adenovirus-based vector. The mechanism takes place at the binding stage and functions independently of CAR. Thus, we have identified a novel binding mechanism whereby adenovirus hijacks HLf, a component of the innate immune system, and uses it as a bridge for attachment to host cells. PMID- 17079303 TI - Replicase genes of murine coronavirus strains A59 and JHM are interchangeable: differences in pathogenesis map to the 3' one-third of the genome. AB - The important roles of the spike protein and other structural proteins in murine coronavirus (MHV) pathogenesis have been demonstrated; however, the role of the replicase gene remains unexplored. We assessed the influence of the replicase genes of the highly neurovirulent MHV-JHM strain and the hepatotropic and mildly neurovirulent A59 strain in acute infection of the mouse. Analysis of chimeric A59/JHM recombinant viruses indicates that the replicase genes are interchangeable and that it is the 3' end of the genome, encoding the structural proteins, rather than the replicase gene, that determines the pathogenic properties of these chimeras. PMID- 17079304 TI - The varicella-zoster virus (VZV) ORF9 protein interacts with the IE62 major VZV transactivator. AB - The varicella-zoster virus (VZV) ORF9 protein is a member of the herpesvirus UL49 gene family but shares limited identity and similarity with the UL49 prototype, herpes simplex virus type 1 VP22. ORF9 mRNA is the most abundantly expressed message during VZV infection; however, little is known concerning the functions of the ORF9 protein. We have found that the VZV major transactivator IE62 and the ORF9 protein can be coprecipitated from infected cells. Yeast two-hybrid analysis localized the region of the ORF9 protein required for interaction with IE62 to the middle third of the protein encompassing amino acids 117 to 186. Protein pull down assays with GST-IE62 fusion proteins containing N-terminal IE62 sequences showed that amino acids 1 to 43 of the acidic transcriptional activation domain of IE62 can bind recombinant ORF9 protein. Confocal microscopy of transiently transfected cells showed that in the absence of other viral proteins, the ORF9 protein was localized in the cytoplasm while IE62 was localized in the nucleus. In VZV-infected cells, the ORF9 protein was localized to the cytoplasm whereas IE62 exhibited both nuclear and cytoplasmic localization. Cotransfection of plasmids expressing ORF9, IE62, and the viral ORF66 kinase resulted in significant colocalization of ORF9 and IE62 in the cytoplasm. Coimmunoprecipitation experiments with antitubulin antibodies indicate the presence of ORF9-IE62-tubulin complexes in infected cells. Colocalization of ORF9 and tubulin in transfected cells was visualized by confocal microscopy. These data suggest a model for ORF9 protein function involving complex formation with IE62 and possibly other tegument proteins in the cytoplasm at late times in infection. PMID- 17079305 TI - Mouse hepatitis virus does not induce Beta interferon synthesis and does not inhibit its induction by double-stranded RNA. AB - Mouse hepatitis virus (MHV) does not induce interferon (IFN) production in fibroblasts or bone marrow-derived dendritic cells. In this report, we show that the essential IFN-beta transcription factors NF-kappaB and IFN regulatory factor 3 are not activated for nuclear translocation and gene induction during infection. However, MHV was unable to inhibit the activation of these factors and subsequent IFN-beta production induced by poly(I:C). Further, MHV infection did not inhibit IFN-beta production mediated by known host pattern recognition receptors (PRRs) (RIG-I, Mda-5, and TLR3). These results are consistent with the notion that double-stranded RNA, produced during MHV infection, is not accessible to cellular PRRs. PMID- 17079306 TI - White spot syndrome virus annexes a shrimp STAT to enhance expression of the immediate-early gene ie1. AB - Although the Janus kinase-signal transducer and activator of transcription (JAK STAT) signaling pathway is part of the antiviral response in arthropods such as Drosophila, here we show that white spot syndrome virus (WSSV) uses a shrimp STAT as a transcription factor to enhance viral gene expression in host cells. In a series of deletion and mutation assays using the WSSV immediate-early gene ie1 promoter, which is active in shrimp cells and also in insect Sf9 cells, an element containing a STAT binding motif was shown to be important for the overall level of WSSV ie1 promoter activity. In the Sf9 insect cell line, a specific protein-DNA complex was detected by using electrophoresis mobility shift assays (EMSA) with the 32P-labeled STAT binding motif of the WSSV ie1 promoter as the probe. When recombinant Penaeus monodon STAT (rPmSTAT) was overexpressed in Sf9 cells, EMSA with specific antibodies confirmed that the STAT was responsible for the formation of the specific protein-DNA complex. Another EMSA showed that in WSSV-infected P. monodon, levels of activated PmSTAT were higher than in WSSV free P. monodon. A transactivation assay of the WSSV ie1 promoter demonstrated that increasing the level of rPmSTAT led to dose-dependent increases in ie1 promoter activity. These results show that STAT directly transactivates WSSV ie1 gene expression and contributes to its high promoter activity. We conclude that WSSV successfully annexes a putative shrimp defense mechanism, which it uses to enhance the expression of viral immediate-early genes. PMID- 17079307 TI - Role of V1V2 and other human immunodeficiency virus type 1 envelope domains in resistance to autologous neutralization during clade C infection. AB - Biologically functional clade C envelope (Env) glycoproteins from the chronically (donor) and newly (recipient) infected partners of four heterosexual transmission pairs in Zambia were cloned and characterized previously. In each case, the donor viral quasispecies contained Envs that were resistant to autologous neutralization by contemporaneous plasma, while the recipient Envs were sensitive to neutralizing antibodies in this donor plasma sample. The donor Envs also varied in length, glycosylation, and amino acid sequence of the V1V2 hypervariable domain of gp120, while the recipient Envs were much more homogeneous. To assess the contribution of V1V2 to the neutralization phenotype of the donor Envs, V1V2 domains from neutralization-sensitive recipient Envs were replaced with donor V1V2 domains, and the autologous neutralization sensitivities of the chimeric Envs were evaluated using a virus-pseudotyping assay. Long donor V1V2 domains regulated sensitivity to autologous neutralization, although the effect was dependent on the Env background. Short donor V1V2 domains did not confer neutralization resistance. Primary sequence differences in V2 were also found to influence neutralization sensitivity in one set of recipient Envs. The results demonstrate that expansion of the V1V2 domain is one pathway to escape from autologous neutralization in subtype C Envs. However, V1V2-independent mechanisms of resistance also exist, suggesting that escape is multifaceted in chronic subtype C infection. PMID- 17079308 TI - Herpesvirus interference with major histocompatibility complex class II restricted T-cell activation. PMID- 17079309 TI - Vaccination of cats with attenuated feline immunodeficiency virus proviral DNA vaccine expressing gamma interferon. AB - A feline immunodeficiency virus (FIV) provirus with a vif gene deletion (FIVDelta vifATGgamma) that coexpresses feline gamma interferon (IFN-gamma) was tested as a proviral DNA vaccine to extend previous studies showing efficacy with an FIV pPPRDelta vif DNA vaccine. Cats were vaccinated with either FIVDelta vifATGgamma or FIV-pPPRDelta vif proviral plasmid DNA or with both FIV-pPPRDelta vif DNA and a feline IFN-gamma expression plasmid (pCDNA-IFNgamma). A higher frequency of FIV specific T-cell proliferation responses was observed in cats immunized with either FIVDelta vifATGgamma or FIV-pPPRDelta vif plus pCDNA-IFNgamma, while virus specific cytotoxic-T-lymphocyte responses were comparable between vaccine groups. Antiviral antibodies were not observed postvaccination. Virus-specific cellular and humoral responses were similar between vaccine groups after challenge with a biological FIV isolate (FIV-PPR) at 13 weeks postimmunization. All vaccinated and unvaccinated cats were infected after FIV-PPR challenge and exhibited similar plasma virus loads. Accordingly, inclusion of plasmids containing IFN-gamma did not enhance the efficacy of FIV-pPPRDelta vif DNA immunization. Interestingly, the lack of protection associated with FIV-pPPRDelta vif DNA immunization contrasted with findings from a previous study and suggested that multiple factors, including timing of FIV-pPPRDelta vif inoculations and challenge, as well as route of challenge virus delivery, may significantly impact vaccine efficacy. PMID- 17079311 TI - Visualization of the interaction between the precursors of VPg, the viral protein linked to the genome of turnip mosaic virus, and the translation eukaryotic initiation factor iso 4E in Planta. AB - The RNA genome of Turnip mosaic virus is covalently linked at its 5' end to a viral protein known as VPg. This protein binds to the translation eukaryotic initiation factor iso 4E [eIF(iso)4E]. This interaction has been shown to be important for virus infection, although its exact biological function(s) has not been elucidated. In this study, we investigated the subcellular site of the VPg eIF(iso)4E interaction using bimolecular fluorescence complementation (BiFC). As a first step, eIF(iso)4E, 6K-VPg-Pro, and VPg-Pro were expressed as full-length green fluorescent protein (GFP) fusions in Nicotiana benthamiana, and their subcellular localizations were visualized by confocal microscopy. eIF(iso)4E was predominantly associated with the endoplasmic reticulum (ER), and VPg-Pro was observed in the nucleus and possibly the nucleolus, while 6K-VPg-Pro-GFP induced the formation of cytoplasmic vesicles budding from the ER. In BiFC experiments, reconstituted green fluorescence was observed throughout the nucleus, with a preferential accumulation in subnuclear structures when the GFP split fragments were fused to VPg-Pro and eIF(iso)4E. On the other hand, the interaction of 6K VPg-Pro with eIF(iso)4E was observed in cytoplasmic vesicles embedded in the ER. These data suggest that the association of VPg with the translation factor might be needed for two different functions, depending of the VPg precursor involved in the interaction. VPg-Pro interaction with eIF(iso)4E may be involved in perturbing normal cellular functions, while 6K-VPg-Pro interaction with the translation factor may be needed for viral RNA translation and/or replication. PMID- 17079310 TI - Evidence for persistent, occult infection in neonatal macaques following perinatal transmission of simian-human immunodeficiency virus SF162P3. AB - To model human immunodeficiency virus (HIV) perinatal transmission, we studied infection of simian-human immunodeficiency virus (SHIV) SF162P3 in 10 pregnant Macaca nemestrina females and their offspring. Four of nine infants born to and suckled by these dams had evidence of infection, a transmission rate of 44.4% (95% confidence interval, 13.7% to 78.8%). We quantified transplacentally acquired and de novo Env-specific immunoglobulin G (IgG), IgM, and neutralizing antibodies in newborns. Transmission of escape variants was confirmed. In utero infection (n = 1) resulted in high viremia, depletion of peripheral CD4+ T cells, and rapid evolution of env in blood and tissues. Peripartum or postpartum SHIV infection (n = 3) resulted in postacute viral control that was undetectable by very sensitive multiplex PCR, despite increasing antibodies. Seropositive infants with highly controlled viremia had homogeneous peripheral blood env sequences, and their tissues had <3 copies per million cells. A high incidence of seropositive virus-low or -negative SHIV infection in infant macaques has implications for HIV type 1 perinatal transmission and detection. PMID- 17079312 TI - Borna disease virus matrix protein is an integral component of the viral ribonucleoprotein complex that does not interfere with polymerase activity. AB - We have recently shown that the matrix protein M of Borna disease virus (BDV) copurifies with the affinity-purified nucleoprotein (N) from BDV-infected cells, suggesting that M is an integral component of the viral ribonucleoprotein complex (RNP). However, further studies were hampered by the lack of appropriate tools. Here we generated an M-specific rabbit polyclonal antiserum to investigate the intracellular distribution of M as well as its colocalization with other viral proteins in BDV-infected cells. Immunofluorescence analysis revealed that M is located both in the cytoplasm and in nuclear punctate structures typical for BDV infection. Colocalization studies indicated an association of M with nucleocapsid proteins in these nuclear punctate structures. In situ hybridization analysis revealed that M also colocalizes with the viral genome, implying that M associates directly with viral RNPs. Biochemical studies demonstrated that M binds specifically to the phosphoprotein P but not to N. Binding of M to P involves the N terminus of P and is independent of the ability of P to oligomerize. Surprisingly, despite P-M complex formation, BDV polymerase activity was not inhibited but rather slightly elevated by M, as revealed in a minireplicon assay. Thus, unlike M proteins of other negative-strand RNA viruses, BDV-M seems to be an integral component of the RNPs without interfering with the viral polymerase activity. We propose that this unique feature of BDV-M is a prerequisite for the establishment of BDV persistence. PMID- 17079313 TI - Prion interference is due to a reduction in strain-specific PrPSc levels. AB - When two prion strains infect a single host, one strain can interfere with the ability of the other to cause disease but it is not known whether prion replication of the second strain is also diminished. To further investigate strain interference, we infected hamsters in the sciatic nerve with the long incubation-period transmissible mink encephalopathy (TME) agent DY TME prior to superinfection of hamsters with the short-incubation-period HY TME agent. Increases in the interval between TME agent inoculations resulted in an extension of the incubation period of HY TME or a complete block of the ability of the HY TME agent to cause disease. The sciatic nerve route of inoculation gave the two TME strains access to the same population of neurons, allowing for the potential of prion interference in the lumbar spinal cord. The ability of the DY TME agent to extend the incubation period of HY TME corresponds with detection of DY TME PrP(Sc), the abnormal isoform of the prion protein, in the lumbar spinal cord. The increased incubation period of HY TME or the inability of the HY TME agent to cause disease in the coinfected animals corresponds with a reduction in the abundance of HY TME PrP(Sc) in the lumbar spinal cord. When the two strains were not directed to the same populations of neurons within the lumbar spinal cord, interference between HY TME and DY TME did not occur. This suggests that DY TME agent replication interferes with HY TME agent replication when the two strains infect a common population of neurons. PMID- 17079314 TI - Structure of the 5' nontranslated region of the coxsackievirus b3 genome: Chemical modification and comparative sequence analysis. AB - Coxsackievirus B3 (CVB3) is a picornavirus which causes myocarditis and pancreatitis and may play a role in type I diabetes. The viral genome is a single 7,400-nucleotide polyadenylated RNA encoding 11 proteins in a single open reading frame. The 5' end of the viral genome contains a highly structured nontranslated region (5'NTR) which folds to form an internal ribosome entry site (IRES) as well as structures responsible for genome replication, both of which are critical for virulence. A structural model of the CVB3 5'NTR, generated primarily by comparative sequence analysis and energy minimization, shows seven domains (I to VII). While this model provides a preliminary basis for structural analysis, the model lacks comprehensive experimental validation. Here we provide experimental evidence from chemical modification analysis to determine the structure of the CVB3 5'NTR. Chemical probing results show that the theoretical model for the CVB3 5'NTR is largely, but not completely, supported experimentally. In combination with our chemical probing data, we have used the RNASTRUCTURE algorithm and sequence comparison of 105 enterovirus sequences to provide evidence for novel secondary and tertiary interactions. A comprehensive examination of secondary structure is discussed, along with new evidence for tertiary interactions. These include a loop E motif in domain III and a long-range pairing interaction that links domain II to domain V. The results of our work provide mechanistic insight into key functional elements in the cloverleaf and IRES, thereby establishing a base of structural information from which to interpret experiments with CVB3 and other picornaviruses. PMID- 17079315 TI - Lethal infection of K18-hACE2 mice infected with severe acute respiratory syndrome coronavirus. AB - The severe acute respiratory syndrome (SARS), caused by a novel coronavirus (SARS CoV), resulted in substantial morbidity, mortality, and economic losses during the 2003 epidemic. While SARS-CoV infection has not recurred to a significant extent since 2003, it still remains a potential threat. Understanding of SARS and development of therapeutic approaches have been hampered by the absence of an animal model that mimics the human disease and is reproducible. Here we show that transgenic mice that express the SARS-CoV receptor (human angiotensin-converting enzyme 2 [hACE2]) in airway and other epithelia develop a rapidly lethal infection after intranasal inoculation with a human strain of the virus. Infection begins in airway epithelia, with subsequent alveolar involvement and extrapulmonary virus spread to the brain. Infection results in macrophage and lymphocyte infiltration in the lungs and upregulation of proinflammatory cytokines and chemokines in both the lung and the brain. This model of lethal infection with SARS-CoV should be useful for studies of pathogenesis and for the development of antiviral therapies. PMID- 17079316 TI - Small interfering RNA targeted to hepatitis C virus 5' nontranslated region exerts potent antiviral effect. AB - Hepatitis C virus (HCV) is a major cause of cirrhosis and hepatocellular carcinoma. Interferon alone or together with ribavirin is the only therapy for HCV infection; however, a significant number of HCV-infected individuals do not respond to this treatment. Therefore, the development of new therapeutic options against HCV is a matter of urgency. In the present study, we have examined vectors carrying short hairpin RNA (shRNA) targeting the 5' nontranslated conserved region of the HCV genome for inhibition of virus replication. Initially, three sequences were selected, and all three shRNAs (psh-53, psh-274, and psh-375) suppressed HCV internal ribosome entry site (IRES)-mediated translation to different degrees in Huh-7 cells. Next, we introduced siRNA into Huh-7.5 cells persistently infected with HCV genotype 2a (JFH1). The most efficient inhibition of JFH1 replication was observed with psh-274, targeted to the portion from subdomain IIId to IIIe of the IRES. Subsequently, Huh-7.5 cells stably expressing psh-274 further displayed a significant reduction in HCV JFH1 replication. The effect of psh-274 on cell-culture-grown HCV genotype 1a (H77) was also evaluated, and inhibition of virus replication and infectivity titers was observed. In the absence of a cell-culture-grown HCV genotype 1b, the effects of psh-274 on subgenomic and full-length replicons were examined, and efficient inhibition of genome replication was observed. Therefore, we have identified a conserved sequence targeted to the HCV genome that can inhibit replication of different genotypes, suggesting the potential of siRNA as an additional therapeutic modality against HCV infection. PMID- 17079317 TI - Importance of receptor usage, Fli1 activation, and mouse strain for the stem cell specificity of 10A1 murine leukemia virus leukemogenicity. AB - Murine leukemia viruses (MuLV) induce leukemia through a multistage process, a critical step being the activation of oncogenes through provirus integration. Transcription elements within the long terminal repeats (LTR) are prime determinants of cell lineage specificity; however, the influence of other factors, including the Env protein that modulates cell tropism through receptor recognition, has not been rigorously addressed. The ability of 10A1-MuLV to use both PiT1 and PiT2 receptors has been implicated in its induction of blast cell leukemia. Here we show that restricting receptor usage of 10A1-MuLV to PiT2 results in loss of blast cell transformation capacity. However, the pathogenicity was unaltered when the env gene is exchanged with Moloney MuLV, which uses the Cat1 receptor. Significantly, the leukemic blasts express erythroid markers and consistently contain proviral integrations in the Fli1 locus, a target of Friend MuLV (F-MuLV) during erythroleukemia induction. Furthermore, an NB-tropic variant of 10A1 was unable to induce blast cell leukemia in C57BL/6 mice, which are also resistant to F-MuLV transformation. We propose that 10A1- and F-MuLV actually induce identical (erythro)blastic leukemia by a mechanism involving Fli1 activation and cooperation with inherent genetic mutations in susceptible mouse strains. Furthermore, we demonstrate that deletion of the Icsbp tumor suppressor gene in C57BL/6 mice is sufficient to confer susceptibility to 10A1-MuLV leukemia induction but with altered specificity. In summary, we validate the significance of the env gene in leukemia specificity and underline the importance of a complex interplay of cooperating oncogenes and/or tumor suppressors in determining the pathogenicity of MuLV variants. PMID- 17079318 TI - Protection against vaccinia virus challenge by CD8 memory T cells resolved by molecular mimicry. AB - Live vaccinia virus (VV) vaccination has been highly successful in eradicating smallpox. However, the mechanisms of immunity involved in mediating this protective effect are still poorly understood, and the roles of CD8 T-cell responses in primary and secondary VV infections are not clearly identified. By applying the concept of molecular mimicry to identify potential CD8 T-cell epitopes that stimulate cross-reactive T cells specific to lymphocytic choriomeningitis virus (LCMV) and VV, we identified after screening only 115 peptides two VV-specific immunogenic epitopes that mediated protective immunity against VV. An immunodominant epitope, VV-e7r130, did not generate cross-reactive T-cell responses to LCMV, and a subdominant epitope, VV-a11r198, did generate cross-reactive responses to LCMV. Infection with VV induced strong epitope specific responses which were stable into long-term memory and peaked at the time virus was cleared, consistent with CD8 T cells assisting in the control of VV. Two different approaches, direct adoptive transfer of VV-e7r-specific CD8 T cells and prior immunization with a VV-e7r-expressing ubiquitinated minigene, demonstrated that memory CD8 T cells alone could play a significant role in protective immunity against VV. These studies suggest that exploiting cross reactive responses between viruses may be a useful tool to complement existing technology in predicting immunogenic epitopes to large viruses, such as VV, leading to a better understanding of the role CD8 T cells play during these viral infections. PMID- 17079319 TI - Bicistronic woodchuck hepatitis virus core and gamma interferon DNA vaccine can protect from hepatitis but does not elicit sterilizing antiviral immunity. AB - The immunity elicited against nucleocapsid of hepatitis B virus (HBV) and closely related woodchuck hepatitis virus (WHV) has been shown to be important in resolution of hepatitis and protection from infection. Further, activity of gamma interferon (IFN-gamma), which may directly inhibit hepadnavirus replication, promotes antiviral defense and favors T helper cell type 1 (Th1) response, which is seemingly a prerequisite of HBV clearance. In this study, to enhance induction of protective immunity against hepadnavirus, healthy woodchucks were immunized with a bicistronic DNA vaccine carrying WHV core (WHc) and woodchuck IFN-gamma (wIFN-gamma) gene sequences. Three groups, each group containing three animals, were injected once or twice with 0.5 mg, 0.9 mg, or 1.5 mg per dose of this vaccine. In addition, four animals received two injections of 0.6 mg or 1 mg WHc DNA alone. All animals were challenged with WHV. The results showed that four of nine animals injected with the bicistronic vaccine and one of four immunized with WHc DNA became protected from serologically evident infection and hepatitis. This protection was not linked to induction of WHc antigen-specific antibodies or T cell proliferative response and was not associated with enhanced transcription of Th1 cytokines or 2',5'-oligoadenylate synthetase. Strikingly, all animals protected from hepatitis became reactive for WHV DNA and carried low levels of replicating virus in hepatic and lymphoid tissues after challenge with WHV. This study shows that the bicistronic DNA vaccine encoding both hepadnavirus core antigen and IFN-gamma was more effective in preventing hepatitis than that encoding virus core alone, but neither of them could mount sterile immunity against the virus or prevent establishment of occult infection. PMID- 17079320 TI - Role of immunoproteasome catalytic subunits in the immune response to hepatitis B virus. AB - Inhibition of hepatitis B virus (HBV) replication and viral clearance from an infected host requires both the innate and adaptive immune responses. Expression of interferon (IFN)-inducible proteasome catalytic and regulatory subunits correlates with the IFN-alpha/beta- and IFN-gamma-mediated noncytopathic inhibition of HBV in transgenic mice and hepatocytes, as well as with clearance of the virus in acutely infected chimpanzees. The immunoproteasome catalytic subunits LMP2 and LMP7 alter proteasome specificity and influence the pool of peptides available for presentation by major histocompatibility complex class I molecules. We found that these subunits influenced both the magnitude and specificity of the CD8 T-cell response to the HBV polymerase and envelope proteins in immunized HLA-A2-transgenic mice. We also examined the role of LMP2 and LMP7 in the IFN-alpha/beta- and IFN-gamma-mediated inhibition of virus replication using HBV transgenic mice and found that they do not play a direct role in this process. These results demonstrate the ability of the IFN-induced proteasome catalytic subunits to shape the HBV-specific CD8 T-cell response and thus potentially influence the progression of infection to acute or chronic disease. In addition, these studies identify a potential key role for IFN in regulating the adaptive immune response to HBV through alterations in viral antigen processing. PMID- 17079321 TI - Glycoprotein M of herpes simplex virus 1 is incorporated into virions during budding at the inner nuclear membrane. AB - It is widely accepted that nucleocapsids of herpesviruses bud through the inner nuclear membrane (INM), but few studies have been undertaken to characterize the composition of these nascent virions. Such knowledge would shed light on the budding reaction at the INM and subsequent steps in the egress pathway. The present study focuses on glycoprotein M (gM), a type III integral membrane protein of herpes simplex virus 1 (HSV-1) that likely contains eight transmembrane domains. The results indicated that gM localized primarily at the perinuclear region, with especially bright staining near the nuclear membrane (NM). Immunogold electron microscopic analysis indicated that, like gB and gD (M. R. Torrisi et al., J. Virol. 66:554-561, 1992), gM localized within both leaflets of the NM, the envelopes of nascent virions that accumulate in the perinuclear space, and the envelopes of cytoplasmic and mature extracellular virus particles. Indirect immunofluorescence studies revealed that gM colocalized almost completely with a marker of the Golgi apparatus and partially with a marker of the trans-Golgi network (TGN), whether or not these markers were displaced to the perinuclear region during infection. gM was also located in punctate extensions and invaginations of the NM induced by the absence of a viral kinase encoded by HSV-1 U(S)3 and within virions located in these extensions. Our findings therefore support the proposition that gM, like gB and gD, becomes incorporated into the virion envelope upon budding through the INM. The localization of viral glycoproteins and Golgi and TGN markers to a perinuclear region may represent a mechanism to facilitate the production of infectious nascent virions, thereby increasing the amount of infectivity released upon cellular lysis. PMID- 17079322 TI - The ORF7b protein of severe acute respiratory syndrome coronavirus (SARS-CoV) is expressed in virus-infected cells and incorporated into SARS-CoV particles. AB - Coronavirus replication is facilitated by a number of highly conserved viral proteins. The viruses also encode accessory genes, which are virus group specific and believed to play roles in virus replication and pathogenesis in vivo. Of the eight putative accessory proteins encoded by the severe acute respiratory distress syndrome associated coronavirus (SARS-CoV), only two-open reading frame 3a (ORF3a) and ORF7a-have been identified in virus-infected cells to date. The ORF7b protein is a putative viral accessory protein encoded on subgenomic (sg) RNA 7. The ORF7b initiation codon overlaps the ORF7a stop codon in a -1 shifted ORF. We demonstrate that the ORF7b protein is expressed in virus-infected cell lysates and from a cDNA encoding the gene 7 coding region, indicating that the sgRNA7 is bicistronic. The translation of ORF7b appears to be mediated by ribosome leaky scanning, and the protein has biochemical properties consistent with that of an integral membrane protein. ORF7b localizes to the Golgi compartment and is incorporated into SARS-CoV particles. We therefore conclude that the ORF7b protein is not only an accessory protein but a structural component of the SARS-CoV virion. PMID- 17079323 TI - The novel human coronaviruses NL63 and HKU1. PMID- 17079324 TI - Differential induction of type I interferon responses in myeloid dendritic cells by mosquito and mammalian-cell-derived alphaviruses. AB - Dendritic cells (DCs) are an important early target cell for many mosquito-borne viruses, and in many cases mosquito-cell-derived arboviruses more efficiently infect DCs than viruses derived from mammalian cells. However, whether mosquito cell-derived viruses differ from mammalian-cell-derived viruses in their ability to induce antiviral responses in the infected dendritic cell has not been evaluated. In this report, alphaviruses, which are mosquito-borne viruses that cause diseases ranging from encephalitis to arthritis, were used to determine whether viruses grown in mosquito cells differed from mammalian-cell-derived viruses in their ability to induce type I interferon (IFN) responses in infected primary dendritic cells. Consistent with previous results, mosquito-cell-derived Ross River virus (mos-RRV) and Venezuelan equine encephalitis virus (mos-VEE) exhibited enhanced infection of primary myeloid dendritic cells (mDCs) compared to mammalian-cell-derived virus preparations. However, unlike the mammalian-cell derived viruses, which induced high levels of type I IFN in the infected mDC cultures, mos-RRV and mos-VEE were poor IFN inducers. Furthermore, the poor IFN induction by mos-RRV contributed to the enhanced infection of mDCs by mos-RRV. These results suggest that the viruses initially delivered by the mosquito vector differ from those generated in subsequent rounds of replication in the host, not just with respect to their ability to infect dendritic cells but also in their ability to induce or inhibit antiviral type I IFN responses. This difference may have an important impact on the mosquito-borne virus's ability to successfully make the transition from the arthropod vector to the vertebrate host. PMID- 17079325 TI - Nuclear import of the preintegration complex is blocked upon infection by human immunodeficiency virus type 1 in mouse cells. AB - Mouse cells do not support human immunodeficiency virus type 1 (HIV-1) replication because of host range barriers at steps including virus entry, transcription, RNA splicing, polyprotein processing, assembly, and release. The exact mechanisms for the suppression, however, are not completely understood. To elucidate further the barriers against HIV-1 replication in mouse cells, we analyzed the replication of the virus in lymphocytes from human CD4/CXCR4 transgenic mice. Although primary splenocytes and thymocytes allowed the entry and reverse transcription of HIV-1, the integration efficiency of the viral DNA was greatly reduced in these cells relative to human peripheral blood mononuclear cells, suggesting an additional block(s) before or at the point of host chromosome integration of the viral DNA. Preintegration processes were further analyzed using HIV-1 pseudotyped viruses. The reverse transcription step of HIV-1 pseudotyped with the envelope of murine leukemia virus or vesicular stomatitis virus glycoprotein was efficiently supported in both human and mouse cells, but nuclear import of the preintegration complex (PIC) of HIV-1 was blocked in mouse cells. We found that green fluorescent protein (GFP)-labeled HIV-1 integrase, which is known to be important in the nuclear localization of the PIC, could not be imported into the nucleus of mouse cells, in contrast to human cells. On the other hand, GFP-Vpr localized exclusively to the nuclei of both mouse and human cells. These observations suggest that, due to the dysfunction of integrase, the nuclear localization of PIC is suppressed in mouse cells. PMID- 17079326 TI - Disseminated simian varicella virus infection in an irradiated rhesus macaque (Macaca mulatta). AB - We describe correlative clinicopathological/virological findings from a simian varicella virus (SVV)-seronegative monkey that developed disseminated varicella 105 days after gamma-irradiation. Twelve other monkeys in the colony were also irradiated, none of which developed varicella. Before irradiation, sera from the monkey that developed disseminated infection and one asymptomatic monkey were available. Analysis indicated that subclinical reactivation of latent SVV from an asymptomatic irradiated monkey likely led to disseminated varicella in the seronegative irradiated monkey. These findings parallel those from humans with disseminated varicella infection and support the usefulness of SVV infection as a model for human varicella-zoster virus infection, particularly virus reactivation after gamma-irradiation. PMID- 17079327 TI - C5 modulates airway hyperreactivity and pulmonary eosinophilia during enhanced respiratory syncytial virus disease by decreasing C3a receptor expression. AB - Enhanced respiratory syncytial virus disease, a serious pulmonary disorder that affected recipients of an inactivated vaccine against respiratory syncytial virus in the 1960s, has delayed the development of vaccines against the virus. The enhanced disease was characterized by immune complex-mediated airway hyperreactivity and a severe pneumonia associated with pulmonary eosinophilia. In this paper, we show that complement factors contribute to enhanced-disease phenotypes. Mice with a targeted disruption of complement component C5 affected by the enhanced disease displayed enhanced airway reactivity, lung eosinophilia, and mucus production compared to wild-type mice and C5-deficient mice reconstituted with C5. C3aR expression in bronchial epithelial and smooth muscle cells in the lungs of C5-deficient mice was enhanced compared to that in wild type and reconstituted rodents. Treatment of C5-deficient mice with a C3aR antagonist significantly attenuated airway reactivity, eosinophilia, and mucus production. These results indicate that C5 plays a crucial role in modulating the enhanced-disease phenotype, by affecting expression of C3aR in the lungs. These findings reveal a novel autoregulatory mechanism for the complement cascade that affects the innate and adaptive immune responses. PMID- 17079328 TI - Suppression of coronavirus replication by inhibition of the MEK signaling pathway. AB - We previously demonstrated that infection of cultured cells with murine coronavirus mouse hepatitis virus (MHV) resulted in activation of the mitogen activated protein kinase (Raf/MEK/ERK) signal transduction pathway (Y. Cai et al., Virology 355:152-163, 2006). Here we show that inhibition of the Raf/MEK/ERK signaling pathway by the MEK inhibitor UO126 significantly impaired MHV progeny production (a reduction of 95 to 99% in virus titer), which correlated with the phosphorylation status of ERK1/2. Moreover, knockdown of MEK1/2 and ERK1/2 by small interfering RNAs suppressed MHV replication. The inhibitory effect of UO126 on MHV production appeared to be a general phenomenon since the effect was consistently observed in all six different MHV strains and in three different cell types tested; it was likely exerted at the postentry steps of the virus life cycle because the virus titers were similarly inhibited from infected cells treated at 1 h prior to, during, or after infection. Furthermore, the treatment did not affect the virus entry, as revealed by the virus internalization assay. Metabolic labeling and reporter gene assays demonstrated that translation of cellular and viral mRNAs appeared unaffected by UO126 treatment. However, synthesis of viral genomic and subgenomic RNAs was severely suppressed by UO126 treatment, as demonstrated by a reduced incorporation of [3H]uridine and a decrease in chloramphenicol acetyltransferase (CAT) activity in a defective interfering RNA-CAT reporter assay. These findings indicate that the Raf/MEK/ERK signaling pathway is involved in MHV RNA synthesis. PMID- 17079329 TI - The amino-conserved domain of human cytomegalovirus UL80a proteins is required for key interactions during early stages of capsid formation and virus production. AB - Assembly of many spherical virus capsids is guided by an internal scaffolding protein or group of proteins that are often cleaved and eliminated in connection with maturation and incorporation of the genome. In cytomegalovirus there are at least two proteins that contribute to this scaffolding function; one is the maturational protease precursor (pUL80a), and the other is the assembly protein precursor (pUL80.5) encoded by a shorter genetic element within UL80a. Yeast GAL4 two-hybrid assays established that both proteins contain a carboxyl-conserved domain that is required for their interaction with the major capsid protein (pUL86) and an amino-conserved domain (ACD) that is required for their self interaction and for their interaction with each other. In the work reported here, we demonstrate that when the ACD is deleted (deltaACD) or disrupted by a point mutation (L47A), the bacterially expressed mutant protein sediments as a monomer during rate-velocity centrifugation, whereas the wild-type protein sediments mainly as oligomers. We also show that the L47A mutation reduces the production of infectious virus by at least 90%, results in the formation of irregular nuclear capsids, gives rise to tube-like structures in the nucleus that resemble the capsid core in cross-section and contain UL80 proteins, slows nuclear translocation of the major capsid protein, and may slow cleavage by the maturational protease. We provide physical corroboration that mutating the ACD disrupts self-interaction of the UL80 proteins and biological support for the proposal that the ACD has a critical role in capsid assembly and production of infectious virus. PMID- 17079330 TI - Hijacking components of the cellular secretory pathway for replication of poliovirus RNA. AB - Infection of cells with poliovirus induces a massive intracellular membrane reorganization to form vesicle-like structures where viral RNA replication occurs. The mechanism of membrane remodeling remains unknown, although some observations have implicated components of the cellular secretory and/or autophagy pathways. Recently, we showed that some members of the Arf family of small GTPases, which control secretory trafficking, became membrane-bound after the synthesis of poliovirus proteins in vitro and associated with newly formed membranous RNA replication complexes in infected cells. The recruitment of Arfs to specific target membranes is mediated by a group of guanine nucleotide exchange factors (GEFs) that recycle Arf from its inactive, GDP-bound state to an active GTP-bound form. Here we show that two different viral proteins independently recruit different Arf GEFs (GBF1 and BIG1/2) to the new structures that support virus replication. Intracellular Arf-GTP levels increase approximately 4-fold during poliovirus infection. The requirement for these GEFs explains the sensitivity of virus growth to brefeldin A, which can be rescued by the overexpression of GBF1. The recruitment of Arf to membranes via specific GEFs by poliovirus proteins provides an important clue toward identifying cellular pathways utilized by the virus to form its membranous replication complex. PMID- 17079331 TI - Novel blocker of NFAT activation inhibits IL-6 production in human myometrial arteries and reduces vascular smooth muscle cell proliferation. AB - The calcineurin/nuclear factor of activated T cells (NFAT) signaling pathway has been found to play a role in regulating growth and differentiation in several cell types. However, the functional significance of NFAT in the vasculature is largely unclear. Here we show that NFATc1, NFATc3, and NFATc4 are expressed in human myometrial arteries. Confocal immunofluorescence and Western blot analysis revealed that endothelin-1 efficiently increases NFATc3 nuclear accumulation in native arteries. Endothelin-1 also stimulates NFAT-dependent transcriptional activity, as shown by a luciferase reporter assay. Both the agonist-induced NFAT nuclear accumulation and transcriptional activity were prevented by the calcineurin inhibitor CsA and by the novel NFAT blocker A-285222. Chronic inhibition of NFAT significantly reduced IL-6 production in intact myometrial arteries and inhibited cell proliferation in vascular smooth muscle cells cultured from explants from the same arteries. Furthermore, by using small interfering RNA-mediated reduction of NFATc3, we show that this isoform is involved in the regulation of cell proliferation. Protein synthesis in intact arteries was investigated using autoradiography of [(35)S]methionine incorporation in serum-free culture. Inhibition of NFAT signaling did not affect overall protein synthesis or specifically the synthesis rates of major proteins associated with the contractile/cytoskeletal system. An intact contractile phenotype under these conditions was also shown by unchanged force response to depolarization or agonist stimulation. Our results demonstrate NFAT expression and activation in native human vessels and point out A-285222 as a powerful pharmacological blocker of NFAT signaling in the vasculature. PMID- 17079332 TI - Hprt-targeted transgenes provide new insights into smooth muscle-restricted promoter activity. AB - Mouse telokin and SM22alpha promoters have previously been shown to direct smooth muscle cell-specific expression of transgenes in vivo in adult mice. However, the activity of these promoters is highly dependent on the integration site of the transgene. In the current study, we found that the ectopic expression of telokin promoter transgenes could be abolished by flanking the transgene with insulator elements from the H19 gene. However, the insulator elements did not increase the proportion of mouse lines that exhibited consistent, detectable levels of transgene expression. In contrast, when transgenes were targeted to the hprt locus, both telokin and SM22alpha promoters resulted in reproducible patterns and levels of transgene expression in all lines of mice examined. Telokin promoter transgene expression was restricted to smooth muscle tissues in adult and embryonic mice. As reported previously, SM22alpha transgenes were expressed at high levels specifically in arterial smooth muscle cells; however, in contrast to randomly integrated transgenes, the hprt-targeted SM22alpha transgenes were also expressed at high levels in smooth muscle cells in veins, bladder, and gallbladder. Using hprt-targeted transgenes, we further analyzed elements within the telokin promoter required for tissue specific activity in vivo. Analysis of these transgenes revealed that the CArG element in the telokin promoter is required for promoter activity in all tissues and that the CArG element and adjacent AT-rich region are sufficient to drive transgene expression in bladder but not intestinal smooth muscle cells. PMID- 17079333 TI - Phosphorylation of SIMPL modulates RelA-associated NF-kappaB-dependent transcription. AB - Epidemiological data have implicated perturbations in the regulation of NF-kappaB activity to diseases that affect a large number of Americans today. Specifically, chronic activation of genes involved in the inflammatory response is associated with the progression of and complications in diabetes, arthritis, atherosclerosis, and cancer. Insight into the mechanisms governing the regulation of NF-kappaB transcriptional activity will provide the molecular link between NF kappaB and these pathological states. SIMPL (signaling molecule that associates with mouse Pelle-like kinase) is a component of a signaling pathway through which tumor necrosis factor-alpha (TNF-alpha) induces NF-kappaB-controlled gene transcription. SIMPL interacts with the nuclear pool of the NF-kappaB subunit, p65, in a TNF-alpha-dependent manner to enhance p65-dependent gene transcription. How SIMPL activity is regulated is unknown. Under basal as well as TNF-alpha stimulated conditions, SIMPL phosphopeptides were identified. SIMPL mutants lacking sites that are phosphorylated under basal conditions diminished p65 transactivation activity but had no effect on SIMPL nuclear localization. SIMPL mutants lacking sites of TNF-alpha-enhanced phosphorylation impaired nuclear localization and prevented TNF-alpha-induced p65 transactivation activity. Together, these studies reveal that phosphorylation of the SIMPL protein plays a critical role in SIMPL regulation by affecting both SIMPL subcellular localization and the p65 coactivator function of SIMPL. PMID- 17079334 TI - Single membrane tether extraction from adult and neonatal dermal microvascular endothelial cells. AB - Membrane tethers were found to be extracted from leukocytes and macrovascular endothelial cells (e.g., human umbilical vein endothelial cells or HUVECs) when a point pulling force was exerted. These tethers stabilize leukocyte rolling on the endothelium during the inflammatory response. However, little is known about tether extraction from other vascular cells like microvascular endothelial cells (MECs). In this study, we extracted tethers from both adult and neonatal dermal MECs with the micropipette aspiration technique. We found a linear relationship between the pulling force and tether growth velocity for both cell lines. This constitutive relationship is mainly determined by the membrane mechanical property and the underlying actin-based cytoskeleton for both attached and suspended endothelial cells. It is independent of cell surface receptor type, attachment state, cytokine stimulation, or cell lineage. For both types of MECs, the threshold forces are approximately 50 pN and the effective viscosities are around 0.5 pN x s/microm. These results, which are close to what was obtained from HUVECs, indicate that homogeneity is preserved in terms of tether extraction among different types of endothelial cells, and simultaneous tethers are likely extracted when leukocytes roll on either microvascular or macrovascular surfaces. PMID- 17079335 TI - Elaboration of a novel technique for purification of plasma membranes from Xenopus laevis oocytes. AB - Over the past two decades, Xenopus laevis oocytes have been widely used as an expression system to investigate both physiological and pathological properties of membrane proteins such as channels and transporters. Past studies have clearly shown the key implications of mistargeting in relation to the pathogenesis of these proteins. To unambiguously determine the plasma membrane targeting of a protein, a thorough purification technique becomes essential. Unfortunately, available techniques are either too cumbersome, technically demanding, or require large amounts of material, all of which are not adequate when using oocytes individually injected with cRNA or DNA. In this article, we present a new technique that permits excellent purification of plasma membranes from X. laevis oocytes. This technique is fast, does not require particular skills such as peeling of vitelline membrane, and permits purification of multiple samples from as few as 10 and up to >100 oocytes. The procedure combines partial digestion of the vitelline membrane, polymerization of the plasma membrane, and low-speed centrifugations. We have validated this technique essentially with Western blot assays on three plasma membrane proteins [aquaporin (AQP)2, Na(+)-glucose cotransporter (SGLT)1, and transient receptor potential vanilloid (TRPV)5], using both wild-type and mistargeted forms of the proteins. Purified plasma membrane fractions were easily collected, and samples were found to be adequate for Western blot identification. PMID- 17079336 TI - Differential contributions of Shaker and Shab K+ currents to neuronal firing patterns in Drosophila. AB - Different K(+) currents participate in generating neuronal firing patterns. The Drosophila embryonic "giant" neuron culture system has facilitated current- and voltage-clamp recordings to correlate distinct excitability patterns with the underlying K(+) currents and to delineate the mutational effects of identified K(+) channels. Mutations of Sh and Shab K(+) channels removed part of inactivating I(A) and sustained I(K), respectively, and the remaining I(A) and I(K) revealed the properties of their counterparts, e.g., Shal and Shaw channels. Neuronal subsets displaying the delayed, tonic, adaptive, and damping spike patterns were characterized by different profiles of K(+) current voltage dependence and kinetics and by differential mutational effects. Shab channels regulated membrane repolarization and repetitive firing over hundreds of milliseconds, and Shab neurons showed a gradual decline in repolarization during current injection and their spike activities became limited to high-frequency, damping firing. In contrast, Sh channels acted on events within tens of milliseconds, and Sh mutations broadened spikes and reduced firing rates without eliminating any categories of firing patterns. However, removing both Sh and Shal I(A) by 4-aminopyridine converted the delayed to damping firing pattern, demonstrating their actions in regulating spike initiation. Specific blockade of Shab I(K) by quinidine mimicked the Shab phenotypes and converted tonic firing to a damping pattern. These conversions suggest a hierarchy of complexity in K(+) current interactions underlying different firing patterns. Different lineage defined neuronal subsets, identifiable by employing the GAL4-UAS system, displayed different profiles of spike properties and K(+) current compositions, providing opportunities for mutational analysis in functionally specialized neurons. PMID- 17079337 TI - Serotonin facilitates a persistent calcium current in motoneurons of rats with and without chronic spinal cord injury. AB - In the months after spinal cord transection, motoneurons in the rat spinal cord develop large persistent inward currents (PICs) that are responsible for muscle spasticity. These PICs are mediated by low-threshold TTX-sensitive sodium currents (Na PIC) and L-type calcium currents (Ca PIC). Recently, the Na PIC was shown to become supersensitive to serotonin (5-HT) after chronic injury. In the present paper, a similar change in the sensitivity of the Ca PIC to 5-HT was investigated after injury. The whole sacrocaudal spinal cord from acute spinal rats and spastic chronic spinal rats (S2 level transection 2 mo previously) was studied in vitro. Intracellular recordings were made from motoneurons and slow voltages ramps were applied to measure PICs. TTX was used to block the Na PIC. For motoneurons of chronic spinal rats, a low dose of 5-HT (1 microM) significantly lowered the threshold of the Ca PIC from -56.7 +/- 6.0 to -63.1 +/- 7.1 mV and increased the amplitude of the Ca PIC from 2.4 +/- 1.0 to 3.0 +/- 0.73 nA. Higher doses of 5-HT acted similarly. For motoneurons of acute spinal rats, low doses of 5-HT had no significant effects, whereas a high dose (about 30 microM) significantly lowered the threshold of the L-Ca PIC from -58.5 +/- 14.8 to -62.5 +/- 3.6 mV and increased the amplitude of the Ca PIC from 0.69 +/- 1.05 to 1.27 +/- 1.1 nA. Thus Ca PICs in motoneurons are about 30-fold supersensitive to 5-HT in chronic spinal rats. The 5-HT-induced facilitation of the Ca PIC was blocked by nimodipine, not by the I(h) current blocker Cs(+) (3 mM) or the SK current blocker apamin (0.15 microM), and it lasted for hours after the removal of 5-HT from the nCSF, even increasing initially after removing 5-HT. The effects of 5-HT make motoneurons more excitable and ultimately lead to larger, more easily activated plateaus and self-sustained firing. The supersensitivity to 5-HT suggests the small amounts of endogenous 5-HT below the injury in a chronic spinal rat may act on supersensitive receptors to produce large Ca PICs and ultimately enable muscle spasms. PMID- 17079339 TI - Responses of reticulospinal neurons in the lamprey to lateral turns. AB - When swimming, the lamprey maintains a definite orientation of its body in the vertical planes, in relation to the gravity vector, as the result of postural vestibular reflexes. Do the vestibular-driven mechanisms also play a role in the control of the direction of swimming in the horizontal (yaw) plane, in which the gravity cannot be used as a reference direction? In the present study, we addressed this question by recording responses to lateral turns in reticulospinal (RS) neurons mediating vestibulospinal reflexes. In intact lampreys, the activity of axons of RS neurons was recorded in the spinal cord by implanted electrodes. Vestibular stimulation was performed by periodical turns of the animal in the yaw plane (60 degrees peak to peak). It was found that the majority of responding RS neurons were activated by the contralateral turn. By removing one labyrinth, we found that yaw responses in RS neurons were driven mainly by input from the contralateral labyrinth. We suggest that these neurons, when activated by the contralateral turn, will elicit the ipsilateral turn and thus will compensate for perturbations of the rectilinear swimming caused by external factors. It is also known that unilateral eye illumination elicits a contralateral turn in the yaw plane (negative phototaxis). We found that a portion of RS neurons were activated by the contralateral eye illumination. By eliciting an ipsilateral turn, these neurons could mediate the negative phototaxis. PMID- 17079338 TI - Development of spontaneous miniature EPSCs in mouse AVCN neurons during a critical period of afferent-dependent neuron survival. AB - During a critical period prior to hearing onset, cochlea ablation leads to massive neuronal death in the mouse anteroventral cochlear nucleus (AVCN), where cell survival is believed to depend on glutamatergic input. We investigated the development of spontaneous miniature excitatory postsynaptic currents (mEPSCs) in AVCN neurons using whole cell patch-clamp techniques during [postnatal day 7 (P7)] and after (P14, P21) this critical period. We also examined the effects of unilateral cochlea ablation on mEPSC development. The two main AVCN neuron types, bushy and stellate cells, were distinguished electrophysiologically. Bushy cell mEPSCs became more frequent and faster between P7 and P14/P21 but with little change in amplitude. Dendritic filtering of mEPSCs was not detected as indicated by the lack of correlation between 10 and 90% rise times and decay time constants. Seven days after cochlea ablation at P7 or P14, mEPSCs in surviving bushy cells were similar to controls, except that rise and decay times were positively correlated (R = 0.31 and 0.14 for surgery at P7 and P14, respectively). Consistent with this evidence for a shift of synaptic activity from the somata to the dendrites, SV2 staining (a synaptic vesicle marker) forms a ring around somata of control but not experimental bushy cells. In contrast, mEPSCs of stellate cells showed few significant changes over these ages with or without cochlea ablation. Taken together, mEPSCs in mouse AVCN bushy cells show dramatic developmental changes across this critical period, and cochlea ablation may lead to the emergence of excitatory synaptic inputs impinging on bushy cell dendrites. PMID- 17079340 TI - Developmental modulation of the temporal relationship between brain and behavior. AB - Humans and songbirds shape learned vocalizations during a sensorimotor sensitive period or "babbling" phase. The brain mechanisms that underlie the shaping of vocalizations by sensory feedback are not known. We examined song behavior and brain activity in zebra finches during singing as they actively shaped their song toward a tutor model. We now show that the temporal relationship of behavior and activity in the premotor area HVC changes with the development of song behavior. During sensorimotor learning, HVC bursting activity both preceded and followed learned vocalizations by hundreds of milliseconds. Correspondingly, the duration of bursts that occurred during ongoing song motif behavior was prolonged in juveniles, as compared with adults, and was inversely correlated with song maturation. Multielectrode single-unit recording in juveniles revealed that single fast-spiking neurons were active both before and after vocalization. These same neurons responded to auditory stimuli. Collectively, these data indicate that a key aspect of sensory critical periods--prolonged bursting--also applies to sensorimotor development. In addition, prolonged motor discharge and sensory input coincide in single neurons of the developing song system, providing the necessary cellular elements for sensorimotor shaping through activity-dependent mechanisms. PMID- 17079341 TI - Nerve-evoked synchronous release and high K+ -induced quantal events are regulated separately by synaptotagmin I at Drosophila neuromuscular junctions. AB - The distal Ca(2+)-binding domain of synaptotagmin I (Syt I), C2B, has two Ca(2+) binding sites. To study their function in Drosophila, pairs of aspartates were mutated to asparagines and the mutated syt I was expressed in the syt I-null background (P[syt I(B-D1,2N)] and P[syt I(B-D3,4N)]). We examined the effects of these mutations on nerve-evoked synchronous synaptic transmission and high K(+) induced quantal events at embryonic neuromuscular junctions. The P[syt I(B D1,2N)] mutation virtually abolished synaptic transmission, whereas the P[syt I(B D3,4N)] mutation strongly reduced but did not abolish it. The quantal content in P[syt I(B-D3,4N)] increased with the external Ca(2+) concentration, [Ca(2+)](e), with a slope of 1.86 in double-logarithmic plot, whereas that of control was 2.88. In high K(+) solutions the quantal event frequency in P[syt I(B-D3,4N)] increased progressively with [Ca(2+)](e) between 0 and 0.15 mM as in control. In contrast, in P[syt I(B-D1,2N)] the event frequency did not increase progressively between 0 and 0.15 mM and was significantly lower at 0.15 than at 0.05 mM [Ca(2+)](e). The P[syt I(B-D1,2N)] mutation inhibits high K(+)-induced quantal release in a narrow range of [Ca(2+)](e) (negative regulatory function). When Sr(2+) substituted for Ca(2+), nerve-evoked synchronous synaptic transmission was severely depressed and delayed asynchronous release was appreciably increased in control embryos. In high K(+) solutions with Sr(2+), the quantal event frequency was higher than that in Ca(2+) and increased progressively with [Sr(2+)](e) in control and in both mutants. Sr(2+) partially substitutes for Ca(2+) in synchronous release but does not support the negative regulatory function of Syt I. PMID- 17079343 TI - Heterogeneity in the responses of adjacent neurons to natural stimuli in cat striate cortex. AB - When presented with simple stimuli like bars and gratings, adjacent neurons in striate cortex exhibit shared selectivity for multiple stimulus dimensions, such as orientation, direction, and spatial frequency. This has led to the idea that local averaging of neuronal responses provides a more reliable representation of stimulus properties. However, when stimulated with complex, time-varying natural scenes (i.e., movies), striate neurons exhibit highly sparse responses. This raises the question of how much response heterogeneity the local population exhibits when stimulated with movies, and how it varies with separation distance between cells. We investigated this question by simultaneously recording the responses of groups of neurons in cat striate cortex to the repeated presentation of movies using silicon probes in a multi-tetrode configuration. We found, first, that the responses of striate neurons to movies are brief (tens of milliseconds), decorrelated, and exhibit high population sparseness. Second, we found that adjacent neurons differed significantly in their peak firing rates even when they responded to the same frames of a movie. Third, pairs of adjacent neurons recorded on the same tetrodes exhibited as much heterogeneity in their responses as pairs recorded by different tetrodes. These findings demonstrate that complex natural scenes evoke highly heterogeneous responses within local populations, suggesting that response redundancy in a cortical column is substantially lower than previously thought. PMID- 17079342 TI - Neural rate and timing cues for detection and discrimination of amplitude modulated tones in the awake rabbit inferior colliculus. AB - Neural responses to amplitude-modulated (AM) tones in the unanesthetized rabbit inferior colliculus (IC) were studied in an effort to establish explicit relationships between physiological and psychophysical measures of temporal envelope processing. Specifically, responses to variations in modulation depth (m) at the cell's best modulation frequency, with and without modulation maskers, were quantified in terms of average rate and synchronization to the envelope over the entire perceptual dynamic range of depths. Statistically significant variations in the metrics were used to define neural AM detection and discrimination thresholds. Synchrony emerged at modulation depths comparable with psychophysical AM detection sensitivities in some neurons, whereas the lowest rate-based neural thresholds could not account for psychoacoustical thresholds. The majority of rate thresholds (85%) were -10 dB or higher (in 20 log m), and 16% of the population exhibited no systematic dependence of average rate on m. Neural thresholds for AM detection did not decrease systematically at higher SPLs (as observed psychophysically): thresholds remained constant or increased with level for most cells tested at multiple sound-pressure levels (SPLs). At depths higher than the rate-based detection threshold, some rate modulation-depth functions were sufficiently steep with respect to the across-trial variability of the rate to predict depth discrimination thresholds as low as 1 dB (comparable with the psychophysics). Synchrony, on the other hand, did not vary systematically with m in many cells at high modulation depths. A simple computational model was extended to reproduce several features of the modulation frequency and depth dependence of both transient and sustained pure-tone responders. PMID- 17079344 TI - Priming of head premotor circuits during oculomotor preparation. AB - Large, rapid gaze shifts necessitate intricate coordination of the eyes and head. Brief high-frequency bursts of activity within the intermediate and deeper layers of the superior colliculus (dSC) encode desired gaze shifts regardless of component movements of the eyes and head. However, it remains unclear whether low frequency activity emitted by oculomotor neurons within the dSC and elsewhere has any role in eye-head gaze shifts. Here we test the hypothesis that such low frequency activity contributes to eye-head coordination by selectively priming head premotor circuits. We exploited the capacity for short-duration (10 ms, 4 pulses) dSC stimulation to evoke neck muscle responses without compromising ocular stability, stimulating at various intervals of a "gap-saccade" task. Low frequency neural activity in many oculomotor areas (including the dSC) is known to increase during the progression of the gap-saccade task. Stimulation was passed during either a fixation-interval while a central fixation point was illuminated, a 200-ms gap-interval between fixation point offset and target onset, or a movement-interval following target onset. In the two monkeys studied, the amplitude of evoked responses on multiple neck muscles tracked the known increases in low-frequency oculomotor activity during the gap-saccade task, being greater following stimulation passed at the end of the gap- versus the fixation interval, and greater still when the location of stimulation during the movement interval coincided with the area of the dSC generating the ensuing saccade. In one of these monkeys, we obtained a more detailed timeline of how these results co-varied with low-frequency oculomotor activity by stimulating, across multiple trials, at different times within the fixation-, gap- and movement-intervals. Importantly, in both monkeys, baseline levels of neck EMG taken immediately prior to stimulation onset did not co-vary with the known pattern of low-frequency oculomotor activity up until the arrival of a transient burst associated with visual target onset. These baseline results demonstrate that any priming of the head premotor circuits occurs without affecting the output of neck muscle motoneurons, We conclude that low-frequency oculomotor activity primes head premotor circuits well in advance of gaze shift initiation, and in a manner distinct from its effects on the eye premotor circuits. Such distinctions presumably aid the temporal coordination of the eyes and head despite fundamentally different biomechanics. PMID- 17079345 TI - A simple depletion model of the readily releasable pool of synaptic vesicles cannot account for paired-pulse depression. AB - Paired-pulse depression (PPD) is a form of short-term plasticity that plays a central role in processing of synaptic activity and is manifest as a decrease in the size of the response to the second of two closely timed stimuli. Despite mounting evidence to the contrary, PPD is still commonly thought to reflect depletion of the pool of synaptic vesicles available for release in response to the second stimulus. Here it is shown that PPD cannot be accounted for by depletion at excitatory synapses made by hippocampal neurons because PPD is unaffected by changes in the fraction of the readily releasable pool (RRP) released by the first of a pair of pulses. PMID- 17079346 TI - Temporal processing of saccade targets in parietal cortex area LIP during visual search. AB - We studied whether the lateral intraparietal (LIP) area-a subdivision of parietal cortex anatomically interposed between visual cortical areas and saccade executive centers-contains neurons with activity patterns sufficient to contribute to the active process of selecting saccade targets in visual search. Visually responsive neurons were recorded while monkeys searched for a color different target presented concurrently with seven distractors evenly distributed in a circular search array. We found that LIP neurons initially responded indiscriminately to the presentation of a visual stimulus in their response fields, regardless of its feature and identity. Their activation nevertheless evolved to signal the search target before saccade initiation: an ideal observer could reliably discriminate the target from the individual activation of 60% of neurons, on average, 138 ms after stimulus presentation and 26 ms before saccade initiation. Importantly, the timing of LIP neuronal discrimination varied proportionally with reaction times. These findings suggest that LIP activity reflects the selection of both the search target and the targeting saccade during active visual search. PMID- 17079347 TI - Interstitial nucleus of cajal encodes three-dimensional head orientations in Fick like coordinates. AB - Two central, related questions in motor control are 1) how the brain represents movement directions of various effectors like the eyes and head and 2) how it constrains their redundant degrees of freedom. The interstitial nucleus of Cajal (INC) integrates velocity commands from the gaze control system into position signals for three-dimensional eye and head posture. It has been shown that the right INC encodes clockwise (CW)-up and CW-down eye and head components, whereas the left INC encodes counterclockwise (CCW)-up and CCW-down components, similar to the sensitivity directions of the vertical semicircular canals. For the eyes, these canal-like coordinates align with Listing's plane (a behavioral strategy limiting torsion about the gaze axis). By analogy, we predicted that the INC also encodes head orientation in canal-like coordinates, but instead, aligned with the coordinate axes for the Fick strategy (which constrains head torsion). Unilateral stimulation (50 microA, 300 Hz, 200 ms) evoked CW head rotations from the right INC and CCW rotations from the left INC, with variable vertical components. The observed axes of head rotation were consistent with a canal-like coordinate system. Moreover, as predicted, these axes remained fixed in the head, rotating with initial head orientation like the horizontal and torsional axes of a Fick coordinate system. This suggests that the head is ordinarily constrained to zero torsion in Fick coordinates by equally activating CW/CCW populations of neurons in the right/left INC. These data support a simple mechanism for controlling head orientation through the alignment of brain stem neural coordinates with natural behavioral constraints. PMID- 17079348 TI - Long-term potentiation (LTP) in the central amygdala (CeA) is enhanced after prolonged withdrawal from chronic cocaine and requires CRF1 receptors. AB - The amygdala is part of the brain reward circuitry that plays a role in cocaine seeking and abstinence in animals and cocaine craving and relapse in humans. Cocaine-seeking is elicited by cocaine-associated cues, and the basolateral amygdala (BLA) and CeA are essential in forming and communicating drug-related associations that are thought to be critical in long-lasting relapse risk associated with drug addiction. Here we simulated a cue stimulus with high frequency stimulation (HFS) of the BLA-CeA pathway to examine mechanisms that may contribute to drug-related associations. We found enhanced long-term potentiation (LTP) after 14-day but not 1-day withdrawal from 7-day cocaine treatment mediated through N-methyl-d-aspartate (NMDA) receptors (NRs), L-type voltage-gated calcium channels (L-VGCCs), and corticotropin-releasing factor (CRF)(1) receptors; this was accompanied by increased phosphorylated NR1 and CRF(1) protein not associated with changes in NMDA/AMPA ratios in amygdalae from cocaine-treated animals. We suggest that these signaling mechanisms may provide therapeutic targets for the treatment of cocaine cravings. PMID- 17079349 TI - Control of 3D limb dynamics in unconstrained overarm throws of different speeds performed by skilled baseball players. AB - This study investigated how the human CNS organizes complex three-dimensional (3D) ball-throwing movements that require both speed and accuracy. Skilled baseball players threw a baseball to a target at three different speeds. Kinematic analysis revealed that the fingertip speed at ball release was mainly produced by trunk leftward rotation, shoulder internal rotation, elbow extension, and wrist flexion in all speed conditions. The study participants adjusted the angular velocities of these four motions to throw the balls at three different speeds. We also analyzed the dynamics of the 3D multijoint movements using a recently developed method called "nonorthogonal torque decomposition" that can clarify how angular acceleration about a joint coordinate axis (e.g., shoulder internal rotation) is generated by the muscle, gravity, and interaction torques. We found that the study participants utilized the interaction torque to generate larger angular velocities of the shoulder internal rotation, elbow extension, and wrist flexion. To increase the interaction torque acting at these joints, the ball throwers increased muscle torque at the shoulder and trunk but not at the elbow and wrist. These results indicates that skilled ball throwers adopted a hierarchical control in which the proximal muscle torques created a dynamic foundation for the entire limb motion and beneficial interaction torques for distal joint rotations. PMID- 17079350 TI - Encoding of movement dynamics by Purkinje cell simple spike activity during fast arm movements under resistive and assistive force fields. AB - It is controversial whether simple-spike activity of cerebellar Purkinje cells during arm movements encodes movement kinematics like velocity or dynamics like muscle activities. To examine this issue, we trained monkeys to flex or extend the elbow by 45 degrees in 400 ms under resistive and assistive force fields but without altering kinematics. During the task movements after training, simple spike discharges were recorded in the intermediate part of the cerebellum in lobules V-VI, and electromyographic activity was recorded from arm muscles. Velocity profiles (kinematics) in the two force fields were almost identical to each other, whereas not only the electromyographic activities (dynamics) but also simple-spike activities in many Purkinje cells differed distinctly depending on the type of force field. Simple-spike activities encoded much larger mutual information with the type of force field than that with the residual small difference in the height of peak velocity. The difference in simple-spike activities averaged over the recorded Purkinje-cells increased approximately 40 ms before the appearance of the difference in electromyographic activities between the two force fields, suggesting that the difference of simple-spike activities could be the origin of the difference of muscle activities. Simple spike activity of many Purkinje cells correlated with electromyographic activity with a lead of approximately 80 ms, and these neurons had little overlap with another group of neurons the simple-spike activity of which correlated with velocity profiles. These results show that simple-spike activity of at least a group of Purkinje cells in the intermediate part of cerebellar lobules V-VI encodes movement dynamics. PMID- 17079351 TI - Lymphatic vascular density and lymphangiogenesis during tumour progression of carcinoma ex pleomorphic adenoma. AB - AIMS: To assess lymphatic vascular density (LVD) and lymph vessel endothelial proliferation in a series of carcinoma ex pleomorphic adenoma (CXPA) that represents the tumour in the different carcinogenesis phases and tumour progression. METHODS: In 8 cases of early CXPA (intracapsular and minimally invasive tumours), 8 of advanced CXPA (widely invasive tumours) and 10 of pleomorphic adenoma (PA) without malignant transformation, lymphatic vessels and proliferating cells were detected using the antibodies D2-40 and Ki-67 respectively. RESULTS: Comparing early tumours with advanced ones, LVD was not significantly different at the tumour margin. In contrast, regarding intratumoural lymphatics, PA without malignant transformation and early CXPA contained rare, if any, lymph vessels, whereas in widely invasive carcinomas they were more numerous. However, neither intratumoural nor peritumoural LVD were increased in comparison to adjacent normal salivary gland tissue. In no case did dual immunohistochemistry using D2-40 and the cell proliferation marker Ki-67 reveal the existence of proliferating lymphatics. Carcinomatous emboli were found in peritumoural as well as in intratumoural lymphatics only in advanced CXPA without myoepithelial differentiation. CONCLUSION: In CXPA, the lymphatic network is mainly composed of pre-existing lymphatics which are rare in tumours that have not infiltrated outside the confines of the original PA. In the widely invasive CXPA, intratumoural as well as peritumoural lymphatics are a conduit for carcinoma cells, but in carcinomas with myoepithelial differentiation, the neoplastic cells seem to have a lower invasion capacity. PMID- 17079352 TI - Complicated atheromatous plaque as integral atherogenesis. PMID- 17079353 TI - True or not: uncertainty of laboratory results. PMID- 17079354 TI - Epidermal growth factor receptor status in anaplastic thyroid carcinoma. AB - BACKGROUND: The epidermal growth factor receptor (EGFR) has been reported to be overexpressed in anaplastic thyroid carcinoma (ATC). In vitro studies have shown that EGFR tyrosine kinase inhibitors (TKIs) greatly inhibit cellular growth and induced apoptosis in the ATC cell lines, while somatic mutations in the tyrosine kinase domain or an increased gene copy number are associated with increased sensitivity to TKIs in non-small cell lung cancer. AIM: To investigate the prevalence of EGFR overexpression, gene amplification and activating mutation in the tyrosine kinase domain in patients with ATC. METHODS: The EGFR gene status and protein expression were investigated by direct DNA sequencing of the hot-spot regions in exons 18, 19 and 21, fluorescence in situ hybridisation (FISH), and immunohistochemistry in tumour tissues from 23 patients with ATC. RESULTS: On mutational analysis and FISH, neither mutations in the hot-spots nor gene amplification was observed. However, high polysomy was identified in 14/23 (60.9%) patients with ATC. All cases with immunohistochemistry (IHC) positivity (n = 6) had high polysomy, whereas 8/17 (47.1%) cases with IHC negativity had high polysomy (p = 0.048). High polysomy was observed in all 10 cases with giant cell subtype, but in only 4/11 (36.3%) with squamoid and 0/2 with spindle cell sarcomatoid subtype. There was no statistically significant correlation between FISH positivity of ATC tumour and presence of well-differentiated component. CONCLUSION: Despite the low incidence of somatic EGFR gene mutation and amplification in the study samples, in view of the fact that high polysomy was often identified by FISH, as well as the current lack of therapeutic options, EGFR TKIs are worth investigating for treating the patients with ATC who have at least giant cell subtype. PMID- 17079355 TI - Depositing archived paraffin tissue core biopsy specimens in paraffin tissue microarrays using a paraffin tissue punch modified with a countersink. AB - Paraffin tissue microarrays (PTMAs) introduced by Kononen et al in 1998 have become a widely used technique in routine pathology and even more so in research. Kononen used a tissue puncher/arrayer (Beecher Instruments, Sun Prairie, WI, USA) to take paraffin tissue core biopsy specimens (PTCBs) of 0.6-2 mm in diameter from routine paraffin tissue blocks and transfer them to another paraffin block with up to 1000 holes. As pointed out by Mengel et al, however, it is not possible to use the Kononen/Beecher system to construct PTMAs out of archived PTCBs. To overcome this drawback in the extremely popular Beecher system, the paraffin tissue punch was modified by incorporating a conical 4 mm deep countersink. This countersink was milled with a conical precision cutter that can be bought in an ordinary hardware store (cost <5 US dollars). The countersink facilitates the insertion of an archived PTCB into the paraffin tissue punch and enables the construction of PTMAs with previously archived PTCBs using the widely distributed Beecher system. Moreover, this paraffin tissue punch can be used for other systems to create PTMAs, such as the low-budget systems designed by Vogel. PMID- 17079356 TI - Interplay between human papilloma virus infection and p53 gene alterations in head and neck squamous cell carcinoma of an Indian patient population. AB - AIM: To investigate the complex interplay between human papilloma virus (HPV) infection and p53 gene alteration in 92 head and neck squamous cell carcinoma (HNSCC) and 28 leukoplakia samples from eastern India. METHODS: DNA isolated from the patient samples was subjected to HPV detection, loss of heterozygosity (LOH) analysis of the chromosome 17p region harbouring p53, genotyping at the p53 codon 72 locus and sequencing of the entire p53 gene to identify somatic mutations. Codon 72 heterozygotes carrying the p53 mutation were further cloned and resequenced to identify the allele harbouring the mutation. RESULTS: HPV positivity in the HNSCC samples was 69%; 21% of the HNSCC were found to harbour p53 mutations in the coding region of the gene. The absence of the p53 mutation in HPV positive tumours was statistically significant compared to the HPV negative tumours (p = 0.01), but the same did not hold true for p53 LOH (p = 1.0). Among the germline p53 codon 72 heterozygotes, the Pro allele was preferentially lost (p = 0.02) while the Arg allele was mutated in the majority of cases. The risk of HPV mediated tumourigenesis increased with the increase in number of Arg alleles at the codon 72 locus. CONCLUSION: It is proposed that genetic and epigenetic alteration of p53 follow distinct pathways during the development of HNSCC from normal epithelium via dysplasia. The p53 mutation and HPV mediated p53 inactivation possibly constitute two independent pathways of tumourigenesis. PMID- 17079357 TI - Heteroduplex analysis for the three common HFE variants: methodology, reliability and analysis of over 5000 requests for testing. AB - OBJECTIVE: To describe the analysis of over 5300 patient samples for the HFE genotype. METHODS: Blood samples received from hospitals in England, Wales and Ireland were analysed with a single, multiplex PCR using heteroduplex generators for the C282Y, H63D and S65C variants of the HFE gene. PCR products labelled with fluorescent dyes were analysed by capillary electrophoresis. Genotype frequencies were analysed according to the reasons given for testing. RESULTS: Analysis of 400 samples sent in duplicate revealed one error that was associated with reporting rather than the methodology. Of 5327 samples received, 1122 were for family testing, 2470 for diagnostic testing and in 1735 cases no reason was given. Overall, homozygosity for C282Y was found in 14% of samples received for family testing and in 16% of the remaining samples. Clinical indications such as "liver disease" were of little predictive value for homozygosity for C282Y, but this increased if a raised serum ferritin concentration or transferrin saturation was indicated. When the diagnosis was iron overload, 39% of subjects tested were homozygous for C282Y. Compound heterozygosity (C282Y/H63D) was more frequent than in the general population but the frequency was not further increased in subjects for whom there was a diagnosis of iron overload. The frequencies of heterozygosity for H63D or S65C and homozygosity for H63D were not significantly increased in any group compared with the general population frequency. CONCLUSION: These results demonstrate the reliability of the methodology and confirm the difficulty of identifying genetic haemochromatosis purely on the basis of clinical suspicion that haemochromatosis may be responsible for liver disease, diabetes or arthritis. PMID- 17079358 TI - Human hepatic cytochrome p450-specific metabolism of parathion and chlorpyrifos. AB - Organophosphorus pesticides (OPs) remain a potential concern to human health because of their continuing worldwide use. Thiophosphorus OPs, once bioactivated by cytochromes P450 (P450s), form oxon metabolites, which are potent acetylcholinesterase inhibitors. This study investigated the rate of desulfation (activation) and dearylation (detoxification) of parathion and chlorpyrifos in human liver microsomes. In addition, recombinant human P450s were used to quantify, for the first time, the P450-specific kinetic variables (K(m) and V(max)) for each compound for future use in refining human physiologically based pharmacokinetic/pharmacodynamic (PBPK/PD) models of OP exposure. CYP1A2, 2B6, 2C9, 2C19, 3A4, 3A5, and 3A7 were found to be active to a widely varying degree in parathion metabolism, whereas all, with the exception of CYP2C9, were also found to be active in chlorpyrifos metabolism. CYP2B6 and CYP2C19 demonstrated low K(m) and high V(max) values for the metabolism of both model compounds, which supports their role as the primary enzymes that regulate metabolism at low-level human exposures to OPs. With K(m) and V(max) values of 0.61 microM, 4827 pmol/min/nmol P450 and 0.81 microM, 12,544 pmol/min/nmol for formation of paraoxon and chlorpyrifos-oxon, respectively, CYP2B6 favored the desulfation reaction. CYP2C19 activity favored dearylation with K(m) and V(max) values of 0.60 microM, 2338 pmol/min/nmol P450 and 1.63 microM, 13,128 pmol/min/nmol for formation of p-nitrophenol and 3,4,5-tricholorpyrindinol, respectively. P450 specific kinetic parameters for OP metabolism will be used with age-dependent hepatic P450 content to enhance PBPK/PD models so that OP exposures can be modeled to protect human health in different age groups. PMID- 17079359 TI - Recirculatory pharmacokinetic model of the uptake, distribution, and bioavailability of prochlorperazine administered as a thermally generated aerosol in a single breath to dogs. AB - A thermal aerosol generation process is capable of delivering pure drug reliably to the alveoli where it is absorbed systemically. Although deep lung absorption of drugs administered as an aerosol has been shown to be rapid, detailed characterization of their absorption and distribution has not been reported. The present study describes the pharmacokinetics of prochlorperazine from the moment of administration as either a rapid intravenous infusion or a thermally generated aerosol and determines the bioavailability of the aerosol by two independent methods. Prochlorperazine disposition was determined in four anesthetized dogs after a 5-s intravenous infusion and after thermally generated aerosol administration in one breath. Venous blood samples were collected frequently from the time of drug administration to 24 h and left ventricular blood samples were drawn more often until 10 min after drug administration. Prochlorperazine disposition after intravenous and aerosol administration was characterized by fitting a recirculatory model to left ventricular and venous drug concentration data simultaneously. Prochlorperazine aerosol administration produced plasma drug concentrations similar to those after rapid intravenous administration of the same nominal dose, with peak left ventricular concentrations achieved in less than 30 s. Plasma concentration profiles of prochlorperazine administered by both routes were well described by the recirculatory model. Bioavailability of the thermally generated aerosol was consistent and averaged more than 80% of emitted dose. Pulmonary administration of a thermally generated drug aerosol in one breath may be a viable alternative to rapid intravenous administration of drugs requiring rapid and predictable production of effective plasma concentrations. PMID- 17079361 TI - The physiology of bone blood flow: a review. AB - INTRODUCTION: Bone circulation plays an important role in bone physiology, but has been relatively poorly studied, because most techniques of circulatory research are difficult to apply to bone. This article summarizes briefly some of the important aspects of the physiology of bone blood flow most relevant to orthopaedics. METHODS: The gold standard for experimental measurement of bone blood flow is the radioactive microsphere technique, though advances are being made in other techniques, such as positron emission tomography, laser and ultra sound Doppler velocimetry, and near infrared spectroscopy, that may provide useful clinical measurement in the future. RESULTS: Multiple vascular pathways contribute to an adaptive response to traumatic disruption of bone circulation. The microcirculation is not merely a passive conduit for blood flow, but plays an active role in controlling bone processes such as osteochondral ossification. DISCUSSION: The pathophysiology of bone circulation has been associated with osteonecrosis, but more and more evidence is pointing to the importance of bone circulation in fracture repair and osteoporosis, both of which are potentially very exciting areas for future studies. PMID- 17079360 TI - Effect of goldenseal (Hydrastis canadensis) and kava kava (Piper methysticum) supplementation on digoxin pharmacokinetics in humans. AB - Phytochemical-mediated modulation of P-glycoprotein (P-gp) and other drug transporters may give rise to many herb-drug interactions. Serial plasma concentration-time profiles of the P-gp substrate, digoxin, were used to determine whether supplementation with goldenseal or kava kava modified P-gp activity in vivo. Twenty healthy volunteers were randomly assigned to receive a standardized goldenseal (3210 mg daily) or kava kava (1227 mg daily) supplement for 14 days, followed by a 30-day washout period. Subjects were also randomized to receive rifampin (600 mg daily, 7 days) and clarithromycin (1000 mg daily, 7 days) as positive controls for P-gp induction and inhibition, respectively. Digoxin (Lanoxin, 0.5 mg) was administered p.o. before and at the end of each supplementation and control period. Serial digoxin plasma concentrations were obtained over 24 h and analyzed by chemiluminescent immunoassay. Comparisons of area under the curve (AUC)((0-3)), AUC((0-24)), C(max,) CL/F, and elimination half-life were used to assess the effects of goldenseal, kava kava, rifampin, and clarithromycin on digoxin pharmacokinetics. Rifampin produced significant reductions (p < 0.01) in AUC((0-3)), AUC((0-24)), CL/F, t(1/2), and C(max), whereas clarithromycin increased these parameters significantly (p < 0.01). With the exception of goldenseal's effect on C(max) (14% increase), no statistically significant effects on digoxin pharmacokinetics were observed following supplementation with either goldenseal or kava kava. When compared with rifampin and clarithromycin, supplementation with these specific formulations of goldenseal or kava kava did not appear to affect digoxin pharmacokinetics, suggesting that these supplements are not potent modulators of P-gp in vivo. PMID- 17079362 TI - Transport in Haroldfrostland: a metaphor for fluid movement to and through basic multicellular units. PMID- 17079363 TI - Systematic analysis of classification systems for osteonecrosis of the femoral head. AB - BACKGROUND: Multiple classification systems for osteonecrosis of the hip have been developed to assist physicians in the diagnosis and treatment of this potentially debilitating disorder. The purpose of this analysis was to delineate the classification systems utilized in reports published since 1985 and, through a comparison of the most commonly used systems, to identify consistent factors that would allow for cross-publication comparisons to be made. METHODS: We performed a PubMed search for reports of outcome studies concerning treatment methods for osteonecrosis of the hip. All studies of reported outcomes with greater than ten patients were included in the analysis. Various classification systems were tabulated to determine usage frequencies. The four most commonly used systems were then analyzed to determine common factors used for classification. RESULTS: One hundred and fifty-seven studies were available for analysis. Sixteen major classification systems that made use of more than one radiographic factor were identified, and nine of these systems had one to five modifications reported throughout the literature. Additionally, eleven other systems made use of single factors obtained from either magnetic resonance imaging or anatomic data. The review revealed that four classification systems accounted for greater than 85.4% of the reported studies. Parameters for these four systems were stratified to allow for uniformity of patient or study evaluation. CONCLUSIONS: This analysis of the reported classification systems for osteonecrosis of the femoral head revealed several similarities between the most commonly used systems. An analysis of patients can be made with any of the four major systems if specific data are collected according to various magnetic resonance imaging and radiographic findings. This approach will allow for easier comparison of studies across different centers. LEVEL OF EVIDENCE: Prognostic Level III. See Instructions to Authors on jbjs.org for a complete description of levels of evidence. PMID- 17079364 TI - Determining lesion size in osteonecrosis of the femoral head. AB - BACKGROUND: Several studies have documented that the size of the osteonecrotic lesion in the femoral head is an essential parameter in determining prognosis and treatment. There are several methods currently available to measure lesion size, but no general agreement as to which is most useful. In the present study, three different radiographic methods for determining lesion size were evaluated and compared. METHODS: Anteroposterior and lateral radiographs of forty-two hips with osteonecrosis were examined. The extent of osteonecrotic involvement of the femoral head was determined through the use of three different methods: the volume of necrosis by quantitative digital image analysis, and the angular measurements described by Kerboul et al. and Koo and Kim. Graphs were constructed to demonstrate these relationships. RESULTS: Volumetric measurement appeared to be the most reliable. There was only a rough correlation with angular measurements. Several sources of error were noted when simple angular measurements of irregular, three-dimensional lesions were used. The Kerboul method routinely overestimated lesion size and designated 81% of the lesions as "large." The modified Koo and Kim method provided a more even distribution of lesion size and correlated with volumetric measurements in 74% of hips (thirty one of forty-two hips). CONCLUSIONS: Quantitative volumetric measurements appear to be the most reliable method to measure the true size of a three-dimensional osteonecrotic lesion of the femoral head. Volumetric measurement is more accurate than angular measurement and can be performed easily with modern technology. Angular measurements, although somewhat simpler to use than volumetric measurements, may provide only a rough estimate of lesion size, partly due to the considerable differences in outline or location of the necrotic segments. Nevertheless, determination of lesion size must be part of a comprehensive system of staging of this disease, which includes the evaluation of other parameters, such as the extent and degree of articular surface involvement and the status of the hip joint and the acetabulum. LEVEL OF EVIDENCE: Diagnostic Level III. See Instructions to Authors on jbjs.org for a complete description of levels of evidence. PMID- 17079365 TI - Prediction of collapse in femoral head osteonecrosis: a modified Kerboul method with use of magnetic resonance images. AB - BACKGROUND: The hypothesis that the combined necrotic angle measurement from magnetic resonance imaging scans predicts the subsequent risk of collapse in hips with femoral head necrosis was tested. METHODS: Thirty-seven hips with early stage osteonecrosis in thirty-three consecutive patients were investigated. With use of the modified method of Kerboul et al., we measured the arc of the femoral surface involved by necrosis on a midcoronal as well as a midsagittal magnetic resonance image (rather than an anteroposterior and a lateral radiograph) and then calculated the sum of the angles. On the basis of the magnitude of the resulting combined angle, hips were classified into four categories: grade 1 (<200 degrees), grade 2 (200 degrees to 249 degrees), grade 3 (250 degrees to 299 degrees), and grade 4 (>/=300 degrees). After the initial evaluations, the hips were randomly assigned to a core decompression group or a nonoperative group. Patients underwent regular follow-up until femoral head collapse or for a minimum of five years. RESULTS: Seven grade-4 hips and sixteen grade-3 hips had development of femoral head collapse by thirty-six months. Six of the nine grade 2 hips and none of the five grade-1 hips collapsed (log-rank test, p < 0.01). None of the four hips with a combined necrotic angle of /=240 degrees (the high-risk group) collapsed, and four (50%) of the eight hips with a combined necrotic angle between 190 degrees and 240 degrees (the moderate risk group) collapsed during the study period. CONCLUSIONS: The Kerboul combined necrotic angle, as ascertained with use of magnetic resonance imaging scans instead of radiographs, is a good method to assess future collapse in hips with femoral head osteonecrosis. LEVEL OF EVIDENCE: Prognostic Level I. See Instructions to Authors on jbjs.org for a complete description of levels of evidence. PMID- 17079366 TI - Posterior rotational osteotomy for nontraumatic osteonecrosis with extensive collapsed lesions in young patients. AB - BACKGROUND: In young patients with nontraumatic femoral head osteonecrosis with extensive and collapsed lesions, joint preservation is a goal if total joint arthroplasty is to be avoided. We evaluated the effectiveness of a posterior rotational osteotomy in this patient population. METHODS: We reviewed thirty-five hips in twenty-eight young patients with nontraumatic femoral head osteonecrosis treated by posterior femoral neck rotational osteotomy. All femoral heads were collapsed, and seven hips showed joint-space narrowing. Lateral radiographs of the femoral head revealed that 15% of the mean posterior portion and 17% of the mean anterior portion of the femoral head consisted of radiographically apparent living bone. The mean age of the patients (ten women and eighteen men) was twenty eight years. The mean follow-up period was eight years. RESULTS: Less than six months after surgery, the radiographically apparent area of living bone of the femoral head below the acetabular roof was shown to be 59% on the standard anteroposterior radiograph and 54% on the 45 degrees -flexion radiograph. In thirty-three hips (94%), further collapse of the femoral head was prevented and an adequate amount of living bone was demonstrated on the loaded lateral portion of the femoral head on the final follow-up radiographs. Progressive joint-space narrowing was seen in four hips. CONCLUSIONS: In young patients with osteonecrosis and extensively collapsed lesions of the femoral head, posterior femoral neck rotational osteotomy appears to be effective in delaying the progression of degeneration if an adequate area of living bone can be placed under the loaded lateral portion of the acetabulum. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions to Authors on jbjs.org for a complete description of levels of evidence. PMID- 17079367 TI - Survivorship analysis and radiographic outcome following tantalum rod insertion for osteonecrosis of the femoral head. AB - BACKGROUND: For early stages of osteonecrosis, preservation of the femoral head is the primary objective; however, there has not been a consensus on how best to achieve this goal. Core decompression alone is associated with a lack of structural support with inconsistent outcomes, whereas vascularized fibular grafting requires an extensive surgical procedure with high donor-site morbidity and prolonged rehabilitation. The adjunctive use of a porous tantalum implant offers the advantages of core decompression, structural support, minimally invasive surgery, and no donor-site morbidity. The purpose of this study was to assess the survivorship and to evaluate the clinical results and radiographic outcomes of hips in which osteonecrosis of the femoral head was treated with core decompression and a porous tantalum implant. METHODS: We evaluated fifty-four patients (sixty consecutive hips) in whom osteonecrosis of the femoral head was treated with core decompression and insertion of a porous tantalum implant. Fifty two patients (fifty-eight hips) were available for follow-up at a mean of twenty four months. All patients were sixty-five years of age or younger (mean age, thirty-five years). According to the classification system of Steinberg et al., one hip (2%) had stage-I disease, forty-nine hips (84%) had stage-II disease, and eight hips (14%) had stage-III disease. Outcome measures that were used included a limb-specific score (Harris hip score), radiographic outcome measures, and survivorship analysis with revision to total hip arthroplasty as the end point. RESULTS: Overall, nine hips (15.5%) were converted to total hip arthroplasty, including six with stage-II disease and three with stage-III disease. The overall survival rates were 91.8% (95% confidence interval, 87.8% to 95.8%) at twelve months, 81.7% (95% confidence interval, 75.8% to 87.6%) at twenty-four months, and 68.1% (95% confidence interval, 54.7% to 81.5%) at forty-eight months. The absence of chronic systemic diseases resulted in a survival rate of 92% at forty eight months (95% confidence interval, 87.4% to 96.4%). CONCLUSIONS: Treatment of early stage osteonecrosis of the femoral head with core decompression and a porous tantalum implant can be accomplished with a minimally invasive technique and no donor-site morbidity. The early clinical results show encouraging survival rates in patients who do not have chronic systemic disease, especially in association with early stage disease. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions to Authors on jbjs.org for a complete description of levels of evidence. PMID- 17079368 TI - Biophysical stimulation with pulsed electromagnetic fields in osteonecrosis of the femoral head. AB - BACKGROUND: Osteonecrosis of the femoral head is the end point of a disease process that results in bone necrosis, joint edema, and cartilage damage. It leads to joint arthritis that necessitates total hip arthroplasty in many patients. Because of its positive effects on osteogenesis and its chondroprotective effect of articular cartilage, pulsed electromagnetic field stimulation has been proposed as a method to prevent or delay the progression of osteonecrosis. METHODS: A retrospective analysis of the results of treatment with pulsed electromagnetic field stimulation of seventy-six hips in sixty-six patients with osteonecrosis of the femoral head was performed. Patients with Ficat stage I, II, or III osteonecrosis of the femoral head were treated with pulsed electromagnetic field stimulation for eight hours per day for an average of five months. Clinical and diagnostic imaging information was collected at the start of the treatment and at the time of follow-up. The primary end point analyzed was the avoidance of hip surgery, and the secondary end point was limiting the radiographic progression (according to Ficat stage) of osteonecrosis of the femoral head. RESULTS: Fifteen hips required a total hip arthroplasty; twelve of these hips were in patients with Ficat stage-III disease. The need for total hip arthroplasty was significantly higher in patients with Ficat stage-III disease than in patients with Ficat stage-I (p < 0.0001) or II (p < 0.01) disease at the beginning of treatment. Pulsed electromagnetic fields preserved 94% of Ficat stage-I or II hips. Furthermore, radiographic progression (according to Ficat stage) occurred in twenty hips (26%). Pain, present in all patients at the start of the treatment, disappeared after sixty days of stimulation in thirty five patients (53%) and was of moderate intensity in seventeen patients (26%). CONCLUSIONS: The results of this study confirm that pulsed electromagnetic field treatment may be indicated in the early stages of osteonecrosis of the femoral head (Ficat stages I and II). Pulsed electromagnetic field stimulation may be able to either preserve the hip or delay the time until surgery. The authors hypothesize that the short-term effect of pulsed electromagnetic field stimulation may be to protect the articular cartilage from the catabolic effect of inflammation and subchondral bone-marrow edema. The long-term effect of pulsed electromagnetic field stimulation may be to promote osteogenic activity at the necrotic area and prevent trabecular fracture and subchondral bone collapse. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions to Authors on jbjs.org for a complete description of levels of evidence. PMID- 17079369 TI - Bone impaction grafting for corticosteroid-associated osteonecrosis of the knee. AB - BACKGROUND: Osteonecrosis of the distal portion of the femur produces a segment of dead bone in the weight-bearing portion of the femoral condyle, frequently associated with subchondral fracture and collapse and eventually resulting in secondary osteoarthritis. Treatment of these late stages of osteonecrosis in the knee can be problematic. The purpose of the present study was to evaluate a new surgical technique in which the subchondral osteonecrotic lesion is removed. The bone defect is then reconstructed with impacted bone grafts to prevent collapse and/or to regain distal femoral sphericity. METHODS: In this prospective, one surgeon study, nine consecutive knees in six patients were studied, all of which had extensive corticosteroid-associated osteonecrotic lesions of the femoral condyles. Six knees had collapsed lesions when they were initially treated. The mean age of the patients was thirty-one years. Both the clinical and radiographic outcomes were assessed at a minimal follow-up time of two years. RESULTS: At a mean follow-up time of fifty-one months, none of the reconstructed knees had been converted to a total knee prosthesis. The objective Knee Society score improved from a mean of 63 to 89 points. The functional Knee Society score improved from a mean of 19 to 81 points. During the follow-up period, there was no progression of collapse observed; however, three knees showed early signs of osteoarthritis. Clinical success was achieved in six of eight knees, and radiographic success was achieved in seven of nine knees. CONCLUSIONS: At the time of writing (at the time of midterm follow-up), this method appears attractive as a joint-preserving procedure. It is a relatively simple procedure that is not likely to interfere with future knee procedures. It appears that this technique can be effective in knees with collapse of the femoral condyle, and it may delay the need for a total knee arthroplasty. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions to Authors on jbjs.org for a complete description of levels of evidence. PMID- 17079370 TI - Osteonecrosis of the knee after laser or radiofrequency-assisted arthroscopy: treatment with minimally invasive knee arthroplasty. AB - BACKGROUND: Osteonecrosis of the knee after various arthroscopic procedures associated with the use of laser or radiofrequency devices has been described in a few case reports. The purpose of this study was to report on a series of nineteen patients with osteonecrosis of the knee after arthroscopic procedures. A literature search was done to compare this series of patients to previously reported cases. In addition, we analyzed the outcome after treatment with minimally invasive knee arthroplasty. METHODS: We studied patients who had development of osteonecrosis of the knee after a routine arthroscopic procedure. Preoperative and postoperative clinical notes, radiographs, and magnetic resonance images of patients were analyzed. Only those patients with no evidence of osteonecrosis on preoperative magnetic resonance imaging who later had development of osteonecrosis and subsequently required a knee arthroplasty were included. We conducted a search of the current literature to compare the results seen in our patient population with those seen in other patients with this entity. Patients were followed both clinically and radiographically for a mean of sixty-two months. RESULTS: A total of nineteen patients met the inclusion criteria. There were fourteen women and five men with a mean age of sixty-nine years. Six patients underwent an arthroscopy with associated holmium or yttrium aluminum-garnet laser treatment, ten patients had associated radiofrequency treatment, and three patients had microfracture surgery. Subsequent arthroplasty procedures included four unicompartmental knee arthroplasties and fifteen tricompartmental knee arthroplasties. At the time of final follow-up, the mean Knee Society objective score was 95 points. CONCLUSIONS: Arthroscopic procedures may play a role in the development of osteonecrosis of the knee. To our knowledge, this is the largest series of patients to have development of this condition after arthroscopy with associated laser, radiofrequency, or microfracture surgery. The midterm results of knee arthroplasty in this unique patient population are comparable with those of patients undergoing knee arthroplasty for osteoarthritis of the knee. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions to Authors on jbjs.org for a complete description of levels of evidence. PMID- 17079371 TI - Outcomes of total and unicompartmental knee arthroplasty for secondary and spontaneous osteonecrosis of the knee. AB - BACKGROUND: The reported outcomes of patients who underwent total or unicompartmental knee arthroplasty for secondary and spontaneous osteonecrosis of the knee are often from studies that lack the number of subjects necessary to generate meaningful conclusions. We systematically reviewed the available literature in order to define the outcomes of patients after total knee arthroplasty for secondary osteonecrosis and after total or unicompartmental knee arthroplasty for spontaneous osteonecrosis of the knee. METHODS: A literature review yielded twenty cohorts with demographic patient information and outcome scores (global knee scores, radiographic outcomes, and revision rates) for patients who had knee arthroplasty as treatment for either secondary or spontaneous osteonecrosis of the knee. The mean preoperative and postoperative global knee scores, the mean revision rate, and the categorization of the mean "poor" and mean "good" outcomes for the knees with each underlying disease were tabulated and reported. The demographic data and the reported mean global knee scores were weighted by the number of knees in each study. RESULTS: Total knee arthroplasty was performed in 150 knees with secondary osteonecrosis and 148 knees with spontaneous osteonecrosis, and unicompartmental knee arthroplasty was performed in sixty-four knees with spontaneous osteonecrosis. Total knee arthroplasty for spontaneous osteonecrosis of the knee was associated with the best outcomes (higher "good" and postoperative global knee scores and lower revision [3%] and "poor" outcome [6%] rates compared with the other two groups). The outcomes after total knee arthroplasty in knees with secondary osteonecrosis as well as in knees with spontaneous osteonecrosis were better in the cohorts operated on during or after 1985 than in those operated on before 1985. Similarly, the outcomes after unicompartmental knee arthroplasty in knees with spontaneous osteonecrosis of the knee were also better in the cohorts operated on during or after 1985 than in those operated on before 1985. CONCLUSIONS: Total knee arthroplasty performed as treatment for either secondary osteonecrosis or spontaneous osteonecrosis and unicompartmental knee arthroplasty performed as treatment for spontaneous osteonecrosis were associated with improved outcomes in cohorts with more recent operative dates. The evidence suggests that the use of contemporary cemented implants in total knee arthroplasty and the selective use of stems and augments in patients who have development of secondary osteonecrosis after total knee arthroplasty are producing outcomes that are comparable to those seen after total knee arthroplasty for osteoarthritis. Although the outcomes of patients who have total knee arthroplasty for the treatment of spontaneous osteonecrosis of the knee have historically been favorable, such outcomes have also shown particular improvement in the studies from more recent operative periods. Although poor outcomes were seen after unicompartmental knee arthroplasty in earlier studies of patients with spontaneous osteonecrosis of the knee, it is possible that those results were secondary to inappropriate patient selection, as the authors of the most recent and, to our knowledge, the only study to follow established operative indications regarding the use of unicompartmental knee arthroplasty reported excellent results. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions to Authors on jbjs.org for a complete description of levels of evidence. PMID- 17079372 TI - Femoral head viability after Birmingham resurfacing hip arthroplasty: assessment with use of [18F] fluoride positron emission tomography. AB - BACKGROUND: Total hip resurfacing has become increasingly popular over the last decade. There remains concern about the effect of the surgical approach on femoral head viability and the role of resurfacing in the management of established osteonecrosis. In view of these concerns, we examined femoral head viability following resurfacing through a modified anterolateral approach. METHODS: The viability of the femoral heads of ten patients who had undergone successful unilateral Birmingham hip resurfacing was assessed with use of positron emission tomography in conjunction with the injection of fluorine at a mean of twenty months after surgery. For each patient, in both the hip that had undergone resurfacing and the contralateral nonresurfaced hip, activity was measured in four regions of interest: the lateral aspect of the femoral head, the medial aspect of the femoral head, the lateral aspect of the femoral neck, and the proximal aspect of the femur. The uptake of fluorine in each area was converted to standard uptake volumes. RESULTS: No areas of osteonecrosis were seen in the femoral head of any patient. There were no significant differences in the standard uptake volumes as measured in the four regions of the nonresurfaced hips, whereas the median values were higher in all four regions of the resurfaced hips. The difference between the values in the resurfaced hips compared with those in the nonresurfaced hips was only significant (p < 0.05) in the lateral aspect of the femoral head. CONCLUSIONS: This study establishes positron emission tomography in conjunction with injection of fluorine as a possible modality for the assessment of femoral head viability after hip resurfacing. Viability following successful Birmingham hip resurfacing performed through a modified anterolateral approach has also been demonstrated. The increase in bone activity that was seen in the resurfaced hips in our study group may be related to bone remodeling or reperfusion of small areas of osteonecrosis. This technique offers the potential to study femoral head perfusion and viability following all types of resurfacing. LEVEL OF EVIDENCE: Diagnostic Level IV. See Instructions to Authors on jbjs.org for a complete description of levels of evidence. PMID- 17079373 TI - Use of metal-on-metal total hip resurfacing for the treatment of osteonecrosis of the femoral head. AB - BACKGROUND: Recently, with the advent of improved metal-on-metal prostheses, total hip resurfacing has emerged as a viable arthroplasty option. However, it remains controversial whether this procedure should be used in patients with osteonecrosis when the femoral resurfacing component is cemented onto dead bone. The purpose of this study was to analyze the clinical and radiographic outcomes of metal-on-metal total hip resurfacing arthroplasty in patients with osteonecrosis of the femoral head. In addition, this group was compared with a matched group of patients who were diagnosed as having osteoarthritis. METHODS: Forty-two osteonecrotic hips that were treated with a metal-on-metal total hip resurfacing arthroplasty were studied. They were matched by gender, age, prosthesis, surgeon, and surgical approach to forty-two osteoarthritic hips that were treated with the same metal-on-metal prosthesis. In the osteonecrosis group, there were twenty-five men and eleven women, and in the osteoarthritis group, there were twenty-eight men and thirteen women. The mean age at the time of surgery was forty-two years. Patients were followed both clinically and radiographically for a mean of forty-one months. RESULTS: The clinical outcomes were similar for both groups, with a good or excellent outcome in thirty-nine hips (93%) with osteonecrosis and a good or excellent outcome in forty-one hips (98%) with osteoarthritis. In each of the two groups, there were two failures that required conversion to a standard total hip arthroplasty. Survivorship curves were similar for the two patient groups. CONCLUSIONS: The short-term results for metal-on-metal total hip resurfacing for this challenging patient population with osteonecrosis were excellent and comparable with those seen in patients with osteoarthritis. We await long-term results to see if these early results are maintained. LEVEL OF EVIDENCE: Prognostic Level II. See Instructions to Authors on jbjs.org for a complete description of levels of evidence. PMID- 17079374 TI - Metal-on-metal hip resurfacing in osteonecrosis of the femoral head. AB - BACKGROUND: The treatment of end-stage osteonecrosis of the femoral head remains a challenge to the orthopaedic surgeon. Historically, total hip arthroplasty for this condition has been associated with poor rates of survival and function when compared with total hip arthroplasty for the treatment of osteoarthritis. The purpose of this study was to determine the medium-term clinical and radiographic results of metal-on-metal hip resurfacing arthroplasty in patients with end-stage osteonecrosis of the femoral head. METHODS: From June 1994 to March 2004, a consecutive single-surgeon series of seventy-three hip resurfacing procedures were performed in sixty patients for the treatment of end-stage osteonecrosis of the femoral head. The cohort included forty-two men (ten of whom had a bilateral resurfacing) and eighteen women (three of whom had a bilateral resurfacing). The mean age was forty-three years (range, seventeen to sixty-nine years). A clinical and radiographic review was performed. RESULTS: There were four revision operations and one planned revision of the seventy-three hips during the follow up period. Two of these revisions were necessitated by aseptic failure of the femoral component. This represents an overall survival rate of 93.2% at a mean of 6.1 years of follow-up (range, two to twelve years). CONCLUSIONS: On the basis of this study, metal-on-metal resurfacing of the hip for osteonecrosis can be considered a safe and effective form of surgery for this group of patients. Longer-term follow-up is required to confirm the expected continued success of this form of arthroplasty in this difficult-to-treat population. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions to Authors on jbjs.org for a complete description of levels of evidence. PMID- 17079375 TI - Uncemented total hip arthroplasty in young adults with osteonecrosis of the femoral head: a comparative study. AB - BACKGROUND: The outcome of uncemented total hip arthroplasty in patients with osteonecrosis of the femoral head in general, and in young adults in particular, remains largely unknown. This study evaluated the clinical and radiographic results of uncemented total hip arthroplasty in young adults with osteonecrosis of the femoral head and compared these results to those seen in young adults with osteoarthritis. METHODS: Forty-one consecutive patients with osteonecrosis of the femoral head (fifty-two hips) and forty patients with osteoarthritis (fifty-two hips) were treated with an uncemented total hip arthroplasty at our institution. The mean age for the osteonecrosis group was thirty-eight years, and, for the osteoarthritis group, forty-two years. Clinical and radiographic outcomes at a minimum of two years were assessed. RESULTS: At a mean duration of follow-up of three years, the functional improvement was significant in both groups (p < 0.05). The outcome was good to excellent for 94% (forty-nine hips) in the osteonecrosis group and 96% (fifty hips) in the osteoarthritis group. There were two revisions in the osteonecrosis group and one revision in the osteoarthritis group. Survivorship free of revision at the time of the latest follow-up was 96.1% for the osteonecrosis group and 98% for the osteoarthritis group. CONCLUSIONS: The short-term results of cementless total hip arthroplasty in patients with osteonecrosis of the femoral head were encouraging and comparable with the results of a matched group of patients with osteoarthritis. We await further follow-up to see if these promising results hold true. LEVEL OF EVIDENCE: Therapeutic Level III. See Instructions to Authors on jbjs.org for a complete description of levels of evidence. PMID- 17079376 TI - Total hip arthroplasty following failure of free vascularized fibular graft. AB - BACKGROUND: Many treatments for osteonecrosis of the femoral head, including the use of a free vascularized fibular graft, have been advocated in an attempt to delay the need for hip arthroplasty. The purpose of this study was to document the clinical and radiographic results of total hip arthroplasty performed following failure of a free vascularized fibular grafting procedure. METHODS: Twenty total hip arthroplasties in eighteen patients who had previously undergone a free vascularized fibular grafting procedure for the treatment of osteonecrosis were retrospectively reviewed. A straight-stem femoral component was used in twelve hips, a tapered femoral component with removal of residual fibular graft was used in five hips, and a tapered stem without graft removal was used in three hips. The twelve hips with a straight-stem femoral component and previous vascularized fibular grafting were compared with thirty-six osteonecrotic hips in thirty other patients who had undergone total hip arthroplasty but had not had previous free vascularized fibular grafting. The radiographic outcomes with respect to initial femoral component alignment and subsequent migration and the clinical outcomes were compared. RESULTS: Analysis of the immediate postoperative radiographs demonstrated significantly improved alignment of the femoral component when a high-speed burr had been used to remove residual fibular graft (p = 0.001), although doing so did significantly increase both the intraoperative blood loss (p = 0.017) and the operative time (p = 0.0002). There was no significant difference in the amount of migration of either the acetabular or the femoral component between the control and study groups at the time of the most recent follow-up. When comparing patients with or without a previous free vascularized fibular graft, the mean postoperative scores at three years were significantly worse in patients who had undergone a previous free vascularized fibular graft (p = 0.03). One revision occurred in the study group at seventy-two months due to acetabular wear, and one revision occurred in the control group at 108 months due to aseptic loosening. CONCLUSIONS: This study raises concern that the outcome of total hip arthroplasty in patients who previously underwent a free vascularized fibular graft for the treatment of osteonecrosis of the femoral head may be worse than that in patients without previous free vascularized fibular grafting. The intraoperative use of a high-speed burr can improve the alignment of the femoral component by removing more of the residual graft. However, this technique does increase intraoperative blood loss and operative time. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions to Authors on jbjs.org for a complete description of levels of evidence. PMID- 17079377 TI - Use of an alumina-on-alumina bearing system in total hip arthroplasty for osteonecrosis of the hip. AB - BACKGROUND: The results of total hip arthroplasty in patients with osteonecrosis of the femoral head are not always optimal. The use of alumina-on-alumina interfaces in young and active patients may decrease wear and lower the rate of aseptic loosening of the implant and appears to be an attractive alternative to the use of conventional cobalt-chromium-on-polyethylene bearings. The purpose of this study was to evaluate the safety and efficacy of the alumina-on-alumina bearing in patients with osteonecrosis and to compare this group of patients to a group of similarly treated patients with osteoarthritis and a group of patients who received conventional cobalt-chromium-on-polyethylene bearings. METHODS: Patients were selected from a United States Investigational Device Exemption multicenter prospective randomized clinical study that was initiated in 1996. Seventy patients with osteonecrosis of the femoral head (seventy-nine hips) received a cementless alumina-on-alumina bearing system and were directly matched to seventy-six patients with osteoarthritis of the hip (seventy-nine hips) who were managed with the same implant. Both groups were compared with twenty-five patients (twenty-six hips) with osteonecrosis and twenty-five patients (twenty six hips) with osteoarthritis who were managed with a cementless cobalt-chromium on-polyethylene bearing system. All patients received a cementless hydroxyapatite coated femoral stem and were followed both clinically and radiographically. RESULTS: The clinical outcomes for alumina-on-alumina bearings were similar for both osteonecrotic and osteoarthritic hips. The seven-year survival probability was 95.5% for the osteonecrotic hips and 89.4% for the osteoarthritic hips in the alumina-on-alumina bearing group and 92.3% for the osteonecrotic hips and 92.9% for the osteoarthritic hips in the cobalt-chromium-on-polyethylene bearing group. At the time of the most recent follow-up, the mean Harris hip score was 96 points for both the osteonecrotic and the osteoarthritic hips in the alumina-on-alumina group and 96 points for the osteonecrotic hips and 97 points for the osteoarthritic hips in the cobalt-chromium-on-polyethylene bearing group. CONCLUSIONS: The results of the use of alumina-on-alumina and cobalt-chromium-on polyethylene bearings in cementless standard total hip implants in patients with osteonecrosis and osteoarthritis were comparable. The low revision rate for the alumina-on-alumina bearing is encouraging and offers a promising option for younger, more active patients who have this challenging disease. LEVEL OF EVIDENCE: Therapeutic Level III. See Instructions to Authors on jbjs.org for a complete description of levels of evidence. PMID- 17079378 TI - Survivorship of femoral revision hip arthroplasty in patients with osteonecrosis. AB - BACKGROUND: Recent studies have indicated that the rate of successful long-term outcome after primary total hip arthroplasty for patients with osteonecrosis may be improved with the use of later-generation porous-coated prostheses (biologic ingrowth fixation) and cement techniques (cement fixation). Less is known about the long-term outcome after revision arthroplasty in the same patient population. The purpose of this study was to characterize the clinical and radiographic outcomes of revision total hip arthroplasty in patients with osteonecrosis. METHODS: We evaluated thirty-four osteonecrotic hips in thirty patients who had undergone revision of a femoral component of a prior total hip arthroplasty. There were nineteen men (twenty-two hips) and eleven women (twelve hips) with a mean age of forty-six years. Thirty one of thirty-four hips were implanted without cement. The cementless prostheses were of different stem lengths, but thirty of thirty-one were proximally porous coated. The mean duration of follow up was 8.2 years. Prerevision radiographs were used to determine the degree of femoral bone loss according to the classification system of Della Valle and Paprosky. The need for revision was analyzed for correlation to known risk factors for osteonecrosis, age and gender of the patient, and degree of prerevision femoral deficiency. A clinical and radiographic evaluation of outcome was performed. RESULTS: This was the first revision for twenty-seven hips, the second for five hips, and the third for two hips. Preoperatively, the defects included four Type I, nine Type II, fifteen Type IIIA, two Type IIIB, one Type IV, and three unknown. The femoral component was rerevised in twelve of the thirty-four hips. One of the failures was the only fully porous-coated stem that was implanted. One of the three cemented implants failed, as compared with eleven of the thirty-one noncemented implants. Survival rates were 90.9% at five years, 54.8% at ten years, 54.8% at fifteen years, and 27.4% at twenty years. With the small sample size, no relationship could be identified with regard to frequency of re-revision and defects, associated risk factors, patient age, or gender. CONCLUSIONS: There was a high failure rate of revised, uncemented, proximally coated femoral components in patients with osteonecrosis of the femoral head at the time of the intermediate-term follow-up. The cause of failure could not be correlated with patient age, gender, risk factors for osteonecrosis, or femoral bone stock. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions to Authors on jbjs.org for a complete description of levels of evidence. PMID- 17079379 TI - Thrombin peptide TP508 stimulates cellular events leading to angiogenesis, revascularization, and repair of dermal and musculoskeletal tissues. AB - The thrombin peptide, TP508, also known as Chrysalin (OrthoLogic, Tempe, Arizona), is a twenty-three-amino-acid peptide that represents a portion of the receptor-binding domain of the native human thrombin molecule that has been identified as the binding site for a specific class of receptors on fibroblasts and other cells. Preclinical studies with this peptide have shown that it can accelerate tissue repair in both soft and hard tissues by mechanisms that appear to involve up-regulation of genes that initiate a cascade of healing events. These events include recruitment and activation of inflammatory cells, directed migration of cells (chemotaxis), cell proliferation, elaboration of extra cellular matrix, and accelerated revascularization of the healing tissues. Early preclinical dermal wound-healing studies showed that TP508 accelerated healing of both incisional wounds and full-thickness excisional wounds in normal and ischemic skin. In all of these studies, the accelerated healing was associated with increased neovascularization across the incision or in the granulating wound bed. Studies in a rat fracture model have also shown that TP508 accelerates the rate of fracture repair. Gene array analysis of fracture callus from control and TP508-treated fractures indicated that TP508 treatment was associated with an up regulation of early response elements, inflammatory mediators, and genes related to angiogenesis. Similar to what had been seen in dermal wounds, histology from rat fracture callus twenty-one days after treatment indicated that fractures treated with TP508 had significantly more large functional blood vessels than did fractures in the control animals. In vitro studies support these in vivo data and indicate that TP508 may have a direct angiogenic effect by promoting the rate of new vessel growth. The results from phase-1 and phase-2 human clinical studies have shown a positive stimulatory effect of TP508 in the healing of diabetic ulcers and in the repair of fractures to the distal aspect of the radius. Collectively, these studies suggest that TP508 accelerates tissue repair by initiating a cascade of events that lead to an increased rate of tissue revascularization and regeneration. PMID- 17079380 TI - Nonsteroidal anti-inflammatory drug-induced fracture nonunion: an inhibition of angiogenesis? AB - BACKGROUND: Approximately 5% to 10% of fractures may result in delayed union or nonunion. The results of research done over the past three decades have shown that the use of nonsteroidal anti-inflammatory drugs (NSAIDs) has an inhibitory effect on fracture repair, but the exact mechanism of action remains to be elucidated. Cancer research has identified that NSAIDs impede cell proliferation by inhibiting angiogenesis. It is proposed that a similar mechanism occurs in the induction of NSAID-induced nonunions. This hypothesis was investigated in a randomized placebo-controlled trial of the NSAID rofecoxib with use of a murine femoral fracture model. METHODS: Two hundred and forty mice were randomized to receive either the nonsteroidal anti-inflammatory drug rofecoxib (5 mg/kg orally) in a 0.5% methylcellulose solution (the NSAID group) or the 0.5% methylcellulose solution only (the control group). Two hundred and thirty-five of the 240 mice underwent surgery to induce an open transverse middiaphyseal femoral fracture, which was then treated with use of a custom-made external fixator. Five additional animals underwent sham surgery with no fracture induced. Outcomes measures included radiographic assessment, histologic analysis, biomechanical testing, and use of laser Doppler flowmetry to assess blood flow across the fracture gap. RESULTS: Radiography revealed similar healing patterns in both groups; however, at the later stages (day 32), the NSAID group had poorer healing. Histological analysis demonstrated that the control animals healed quicker (at days 24 and 32) and had more callus and less fibrous tissue (at days 8 and 32) than the NSAID animals did. Biomechanical testing found that the control animals were stronger at day 32. Both groups exhibited a similar pattern of blood flow; however, the NSAID group exhibited a lower median flow from day 4 onward (significant at days 4, 16, and 24). Positive correlations were demonstrated between both histological and radiographic assessments of healing and increasing blood flow. NSAID-treated animals exhibited lower blood flow and poorer healing by all parameters. Regression analysis, however, demonstrated that the negative effect of NSAIDs on fracture repair is independent of its inhibitory action on blood flow. CONCLUSIONS: Following the development of a novel method of analyzing functional vascularity across a fracture gap, we have demonstrated that the cyclooxygenase-2 (COX-2) inhibitor rofecoxib has a significant negative effect on blood flow across the fracture gap as well as an inhibiting effect on fracture repair. CLINICAL RELEVANCE: COX-2 inhibitors are marketed as having low side-effect profiles. We propose that these drugs should be used with caution in all patients following osseous trauma and, in particular, after injuries that may already predispose a fracture to a delayed union due to osseous, vascular, or patient-related factors. PMID- 17079381 TI - Alcohol-induced adipogenesis in a cloned bone-marrow stem cell. AB - BACKGROUND: Alcohol has been shown to be associated with osteoporosis and osteonecrosis in patients and in animal models. Recent studies have demonstrated that alcohol contributes to abnormal lipid metabolism in the stromal cells of bone marrow, but the mechanisms have not been defined. The purpose of this study was to evaluate the effects of alcohol on the differentiation of a stem cell that was cloned from bone marrow. METHODS: D1 cells (cloned bone-marrow stem cells from a BALB/c mouse) were treated either with increasing concentrations of ethanol (0.09, 0.15, and 0.21 mol/L) or without alcohol to serve as controls. Morphologic features of the cells were monitored with use of a phase-contrast microscope. Alkaline phosphatase activity was determined with use of a colorimetric assay. The expression of genes that are indicators of adipogenesis [422(aP2), PPARgamma] and osteogenesis (osteocalcin) was evaluated using Northern blot and reverse transcription-polymerase chain reaction assays. RESULTS: The cells treated with ethanol started to accumulate triglyceride vesicles at day seven. The number of adipocytes and the percentage of the area that contained the cells with fat vesicles increased significantly (p < 0.05), and the level of alkaline phosphatase activity diminished with longer durations of exposure to ethanol and with higher concentrations. Analysis of gene expression showed diminished expression of osteocalcin. This occurred without a significant increase in the expression of either the fat-cell-specific gene 422(aP2) or PPARgamma in cells treated with ethanol, suggesting that adipogenesis may occur at a point downstream in the fatty-acid-metabolism pathway. CONCLUSIONS: Alcohol treatment decreases osteogenesis while enhancing adipogenesis in a cloned bone marrow stem cell, indicating that alcohol abuse may be one of the mechanisms leading to osteoporosis and osteonecrosis. This finding explains the clinical observation that there is increased adipogenesis in alcohol-induced osteoporosis and osteonecrosis. CLINICAL RELEVANCE: The inhibition of bone-marrow adipogenesis and the concomitant enhancement of osteogenesis may provide a novel approach to the prevention or treatment of osteonecrosis and osteoporosis. PMID- 17079382 TI - Matrix metalloproteinase-9 induces the formation of cartilage canals in the chondroepiphysis of the neonatal rabbit. AB - BACKGROUND: An investigation of matrix metalloproteinase-9 (MMP-9) and its influence on vascular invasion in the secondary ossification center at the chondroepiphysis of developing long bones was undertaken. The effect of MMP-9 was compared with that of basic fibroblast growth factor (b-FGF), a potent angiogenic factor, and we assessed the chorioallantoic membrane (CAM) culture as a model for angiogenesis in osteochondral tissue. METHODS: Seventy-two femoral and seventy two humeral heads of thirty-six four-day postnatal rabbits were dissected immediately after each animal was killed. Solutions of MMP-9, b-FGF, and phosphate-buffered saline solution were applied, and the femoral and humeral chondroepiphyseal explants were incubated for ten days in CAM culture. This was used as an in vivo model to investigate the growth of blood vessels into the femoral and humeral heads of the neonatal rabbit. The explants were harvested from the CAM culture and analyzed histologically. A three-day incubation was also performed to look for early signs of vascular ingrowth into the cartilage matrix. RESULTS: One hundred and twenty epiphyses from thirty rabbits were placed onto CAM culture successfully; of these, two were harvested at three days to assess early changes and 118 were harvested at ten days. Forty of the 118 cultures were still viable when harvested after ten days, giving a 33% yield. Both MMP-9 and b FGF caused an increased vascular invasion into the chondroepiphysis. New blood vessels derived from the chorioallantoic membrane within cartilage canals were more numerous in MMP-9 treated epiphyses, and larger canals were more commonly seen when compared with a control group. CONCLUSIONS: These findings confirmed that b-FGF is angiogenic at the chondroepiphysis. Matrix metalloproteinase-9 appears to be implicated in vascular invasion and induces the formation of new cartilage canals at the chondroepiphysis. The CAM culture model was a useful model for investigating angiogenesis in osteochondral tissue. CLINICAL RELEVANCE: This study adds to the understanding of the complex biochemical interaction that occurs in cartilage when the advancing vasculature begins growing into the chondroepiphysis. A better knowledge of this angiogenic process will enable a better understanding of the pathological failure or disturbance of vasculogenesis, which results in dysplastic growth disorders and osteonecrosis. PMID- 17079383 TI - Femoral artery constriction by norepinephrine is enhanced by methylprednisolone in a rat model. AB - BACKGROUND: Corticosteroids are associated with femoral head osteonecrosis and arterial hypertension. The patho-mechanism of femoral head osteonecrosis is often attributed to ischemia. The aim of this study was to investigate if corticosteroids directly constrict the femoral artery or if they have a permissive effect on norepinephrine and endothelin-1-induced vasoconstriction. METHODS: Femoral artery segments were harvested from twenty Wistar rats and mounted as ring preparations on a small-vessel myograph for the purpose of making isometric force measurements. For the norepinephrine study, twenty femoral artery segments from ten rats were stimulated cumulatively with norepinephrine before and after incubation with methylprednisolone (5 mug/mL). For the endothelin-1 study, forty femoral artery segments from ten rats were used. The four artery segments from each animal were randomized by pairs to either a corticosteroid treatment group (5 mug/mL methylprednisolone incubation, n = 20) or a control group (placebo incubation, n = 18, as two of the twenty control-group vessels did not meet protocol requirements). Isometric wall tension was plotted and quantified by the EC(50) (the plasma concentration of endothelin-1 required for obtaining 50% of maximal constriction in vivo). RESULTS: In the norepinephrine stimulated group, incubation with methylprednisolone did not directly induce any vasoconstriction but did enhance norepinephrine-elicited vasoconstriction. The norepinephrine dose-response curve displayed a shift to the left after incubation with methylprednisolone. This shift was reflected by a significantly lower mean EC50 of 9.5 x 10(-7) M +/- 5.1 x 10(-7) M after methylprednisolone incubation compared with a mean of 2.5 x 10(-6) M +/- 1.1 x 10(-6) M before incubation (p < 0.005). In the endothelin-1-stimulated group, the endothelin-1 dose-response curve displayed a tendency toward stronger contraction in the vessels that were incubated with methylprednisolone, but this tendency did not reach significance. CONCLUSIONS: Incubation with methylprednisolone enhances norepinephrine-mediated contraction of the femoral artery in a rat model. CLINICAL RELEVANCE: Vasoconstriction of the vascular bed supplying the femoral head can diminish femoral head blood flow, and this may be a factor in the early pathogenesis of corticosteroid-associated femoral head osteonecrosis. PMID- 17079384 TI - Use of genetically engineered bone-marrow stem cells to treat femoral defects: an experimental study. AB - BACKGROUND: Treatment of osteonecrosis continues to be a challenging problem. The replacement of necrotic bone with graft materials that promote osteogenesis and angiogenesis may provide better outcomes for early stage disease. In this study, genetically engineered bone-marrow stem cells were used to enhance repair of a defect in the distal aspect of the femur. METHODS: Cloned bone-marrow stem cells were transfected with traceable genes. Osteoblastic and angiogenic properties of the cells were analyzed. A defect was created bilaterally in the distal portion of the femur of twenty-four mice to mimic a core decompression procedure. The cloned cells were transplanted into each defect of the right femur while the left femur served as control. Bone formation was evaluated radiographically and histomorphometrically. In addition, in twenty-four additional mice, the cells were injected into subcutaneous sites, muscles, and into the renal capsule (eight mice in each group) to evaluate ectopic osteogenesis. RESULTS: Radiopaque tissue appeared two weeks after the cells were transplanted into bone defects and at ectopic sites. Histologic analysis demonstrated that these tissues consisted of newly formed bone from transplanted cells that expressed traceable genes. Four of six bone defects that received cell transplantation were filled with new bone at four weeks, and all of the defects (n = 6) demonstrated complete healing at six weeks. On the control side, complete repair was seen in only two of six bone defects at four weeks and in three of six defects at six weeks. Histomorphometric analysis showed that transplantation of marrow stem cells into bone defects produced more bone at an earlier time-point than occurred in the controls. CONCLUSIONS: This study demonstrated that cloned bone-marrow stem cells can directly form bone after transplantation into bone defects and at ectopic sites, indicating that the in vitro expanded bone-marrow stem cells can serve as a graft material to enhance bone repair and to treat osteonecrosis. CLINICAL RELEVANCE: As an alternative graft material, bone-marrow stem cells may provide new and as yet technologically unachievable solutions to many clinical problems in the areas of musculoskeletal reconstruction and tissue regeneration. PMID- 17079385 TI - Endothelin-1-induced femoral head epiphyseal artery constriction is enhanced by long-term corticosteroid treatment. AB - BACKGROUND: With regard to the pathogenesis of corticosteroid-associated femoral head osteonecrosis, only intravascular and extravascular factors have been discussed. Wall constriction of the lateral epiphyseal arteries, which are the main supplying arteries within the femoral head, has not previously been investigated. This study examined the effect of long-term corticosteroid treatment on vasoconstriction of the lateral epiphyseal arteries of the femoral head in a porcine model. METHODS: The immature female Danish Landrace pig was chosen as the model for our study because femoral head osteonecrosis can be induced in this animal and because the lateral epiphyseal arteries of the femoral heads are of the appropriate size for myographic investigation. Twenty-four female Danish Landrace pigs (two siblings from each of twelve litters for a total of twelve sibling pairs) were divided by randomly separating the sibling pairs into two groups, with a total of twelve pigs in each group. One group of twelve animals received a daily dose of 100 mg of methylprednisolone orally for three months. The other group of twelve pigs served as controls and received no corticosteroids. After the animals were killed with an intravenous injection of pentobarbital, the lateral epiphyseal arteries of the femoral head were isolated from the cancellous bone. These arteries were mounted as ring preparations on a small-vessel myograph for measurement of isometric force development. The effects on the resected vessels of the vasoactive agents endothelin-1, noradrenaline, bradykinin, substance P, and nitrous oxide were investigated. RESULTS: Vasoconstriction induced by increasing doses of endothelin-1 was increased after three months of methylprednisolone treatment compared with the vasoconstriction in vessels from control animals. Sensitivity to noradrenaline was not altered by this treatment. After submaximal precontraction by noradrenaline, vasorelaxation was elicited by substance P, nitrous oxide, and increasing doses of bradykinin without differences between the corticosteroid-treated and the control groups. CONCLUSIONS: Endothelin-1-induced vasoconstriction of the epiphyseal arteries of the femoral head was increased in this corticosteroid-treated pig model. Such vasoconstriction may lead to a reduction in femoral head blood flow and to local ischemia and thus may be a newly described factor in the multifactorial pathomechanism of osteonecrosis of the femoral head. PMID- 17079386 TI - Professionalism in publishing. PMID- 17079387 TI - Functional outcome and risk of recurrent instability after primary traumatic anterior shoulder dislocation in young patients. AB - BACKGROUND: The prevalence and risk factors for recurrent instability and functional impairment following a primary glenohumeral dislocation remain poorly defined in younger patients. We performed a prospective cohort study to evaluate these outcomes. We also aimed to produce guidelines for the design of future clinical trials, assessing the efficacy of interventions designed to improve the outcome after a primary dislocation. METHODS: We performed a prospective cohort study of 252 patients ranging from fifteen to thirty-five years old who sustained an anterior glenohumeral dislocation and were treated with sling immobilization, followed by a physical therapy program. Patients received regular clinical follow up to assess whether recurrent instability had developed. Functional assessments were made and were compared for two subgroups: those who had not had instability develop and those who had received operative stabilization to treat recurrent instability. RESULTS: On survival analysis, instability developed in 55.7% of the shoulders within the first two years after the primary dislocation and increased to 66.8% by the fifth year. The younger male patients were most at risk of instability, and 86.7% of all of the patients known to have recurrent instability had this complication develop within the first two years. A small but measurable degree of functional impairment was present at two years after the initial dislocation in most patients. Sample-size calculations revealed that a relatively small number of patients with a primary dislocation would be required in future clinical trials examining the effects of interventions designed to reduce the prevalence of recurrent instability and improve the functional outcome. CONCLUSIONS: Recurrent instability and deficits of shoulder function are common after primary nonoperative treatment of an anterior shoulder dislocation. There is substantial variation in the risk of instability, with younger males having the highest risk and females having a much lower risk. Future clinical trials to evaluate primary interventions should evaluate the prevalence of recurrent instability and functional deficits, with use of an assessment tool specifically for shoulder instability, during the first two years after the initial dislocation. PMID- 17079388 TI - Revision of a failed patellofemoral arthroplasty to a total knee arthroplasty. AB - BACKGROUND: Patellofemoral arthroplasty is an effective treatment for isolated arthritis of the anterior compartment of the knee, but it is unclear whether it compromises the results of total knee arthroplasty if revision is necessary. The purpose of this study was to review the results of total knee arthroplasty after a prior patellofemoral arthroplasty. METHODS: Twelve failed patellofemoral replacements in ten patients were revised to total knee arthroplasties for isolated progressive tibiofemoral arthritis (six knees), isolated patellofemoral catching and maltracking (three), or a combination of both (three). Revisions were performed at a mean of four years after the patellofemoral arthroplasty. A posterior stabilized implant was used in each knee, without a need for stems, augments, or structural bone graft. Patients were assessed by clinical and radiographic follow-up, including Knee Society clinical and functional scores. Baseline preoperative Knee Society scores were obtained at the time of evaluation of the failed patellofemoral arthroplasty. RESULTS: At a mean follow-up of 3.1 years, the mean Knee Society clinical and functional scores had increased from 57 points preoperatively to 96 points and from 51 points preoperatively to 91 points, respectively. At the most recent follow-up, there was no clinical or radiographic evidence of patellofemoral maltracking, loosening, or wear. CONCLUSIONS: On the basis of our experience in this relatively small series with short-term follow-up, the results of total knee arthroplasty do not seem to be compromised after revision of the failed patellofemoral replacement. PMID- 17079389 TI - Extra-articular distal-third diaphyseal fractures of the humerus. A comparison of functional bracing and plate fixation. AB - BACKGROUND: There are strong advocates for both operative and nonoperative treatment of distal-third diaphyseal fractures of the humerus, but there are few comparative data. We performed a retrospective comparison of these two treatment methods. METHODS: Fifty-one consecutive patients with a closed, extra-articular fracture of the distal one-third of the humeral diaphysis were identified from an orthopaedic trauma database. Forty patients were followed for at least six months or until healing of the fracture. Eleven patients were excluded because of inadequate follow-up. Nineteen patients had been managed with plate-and-screw fixation, and twenty-one had been managed with functional bracing. RESULTS: Among the operatively treated patients, one had loss of fixation, one had a postoperative infection, and one required tendon transfers for the treatment of a preoperative radial nerve palsy that did not resolve. Three new postoperative radial nerve palsies developed, and one had not resolved when the patient was last evaluated, three months after surgery. All operatively treated fractures healed with <10 degrees of angular deformity, and one patient lost 20 degrees of shoulder or elbow motion. Among the nonoperatively treated fractures, two were converted to plate fixation because of the treating surgeons' concern regarding alignment and radial nerve palsy. Only one patient had >30 degrees of malalignment in any plane. Two patients had development of skin breakdown during treatment and completed treatment in a sling. Two patients lost >/=20 degrees of elbow or shoulder motion. CONCLUSIONS: For extra-articular distal-third diaphyseal humeral fractures, operative treatment achieves more predictable alignment and potentially quicker return of function but risks iatrogenic nerve injury and infection and the need for reoperation. Functional bracing can be associated with skin problems and varying degrees of angular deformity, but function and range of motion are usually excellent. PMID- 17079390 TI - Cost-effectiveness of unicompartmental and total knee arthroplasty in elderly low demand patients. A Markov decision analysis. AB - BACKGROUND: Interest in unicompartmental knee arthroplasty has recently increased in the United States, making a firm understanding of the indications for this procedure important. The purpose of this study was to examine the cost effectiveness of unicompartmental knee arthroplasty compared with total knee arthroplasty in elderly low-demand patients. METHODS: A Markov decision model was used to evaluate the cost-effectiveness of unicompartmental knee arthroplasty as compared with total knee arthroplasty in the elderly population. Transition probabilities were estimated from the Norwegian Arthroplasty Register and the arthroplasty literature, and costs were based on the average Medicare reimbursement for unicompartmental, tricompartmental, and revision knee arthroplasties. Outcomes were measured in quality-adjusted life-years. RESULTS: Our model showed unicompartmental knee arthroplasty to be a cost-effective strategy for this population as long as the annual probability of revision is <4%. The cost of unicompartmental knee arthroplasty must be greater than $13,500 or the cost of total knee arthroplasty must be less than $8500 before total knee arthroplasty becomes more cost-effective. CONCLUSIONS: Our model suggests that, on the basis of currently available cost and outcomes data, unicompartmental knee arthroplasty and total knee arthroplasty have similar cost-effectiveness profiles in the elderly low-demand patient population. However, several important parameters that could alter the cost-effectiveness analysis were identified; these included implant survival rates, costs, perioperative mortality and infection rates, and utility values achieved with each procedure. The thresholds identified in this study may help decision-makers to evaluate the cost effectiveness of each strategy as further research characterizes the variables associate with unicompartmental and total knee arthroplasties and may be helpful for designing future appropriate clinical trials. PMID- 17079391 TI - Impact of prostheses on function and quality of life for children with unilateral congenital below-the-elbow deficiency. AB - BACKGROUND: Children with unilateral congenital below-the-elbow deficiency present a dilemma to clinicians. Parents want the child to have a prosthesis and, because it seems that the deficiency will cause functional problems, one is customarily prescribed for infants. Use of the prosthesis is then encouraged throughout childhood. However, these children frequently abandon the prosthesis. There are no evidence-based guidelines regarding prescription of prostheses or standard methods for assessing use and function. METHODS: A multicenter outcomes study was done to assess the quality of life and function of 489 children with a unilateral congenital below-the-elbow deficiency; 321 wore a prosthesis, and 168 did not. The Unilateral Below-the-Elbow Test (UBET) was designed, validated, and administered to these children along with several outcomes measures, including the Pediatric Outcomes Data Collection Instrument (PODCI), the Pediatric Quality of Life Inventory (PedsQL), and the Prosthetic Upper Extremity Functional Index (PUFI). RESULTS: Use of a prosthesis was not associated with any clinically relevant differences in PODCI or PedsQL scores. Non-wearers performed either the same as or better than wearers on the UBET. When queried (with use of the PUFI) about performance of various tasks, non-wearers scored themselves higher than wearers. Children with a unilateral congenital below-the-elbow deficiency scored the same as or higher than the general population on the PedsQL. They scored significantly lower than the general population on the PODCI Upper Extremity Physical Function Domain and higher on the Happiness Domain, but the differences were small. CONCLUSIONS: Prostheses may help with social acceptance or may be useful as tools for specialized activities, but they do not appear to improve function or quality of life, which are nearly normal for children with unilateral congenital below-the-elbow deficiency regardless of whether they wear a prosthesis. These findings call into question the standard practices of fitting infants with prostheses and encouraging young children to wear the prosthesis. PMID- 17079392 TI - Impact of comorbidities on the measurement of health in patients with ankle osteoarthritis. AB - BACKGROUND: Investigators seeking to understand the impact of musculoskeletal disorders commonly use validated outcome instruments to assess the effect of diseases on physical function and quality of life. However, the influence of concomitant systemic or musculoskeletal comorbidities on these scores has not been widely considered in orthopaedic research. The purpose of this study was to determine how morbidity unrelated to the ankle influences the perception of physical function and pain by patients with ankle osteoarthritis. METHODS: Short Form-36 (SF-36) Physical Component Summary (PCS) and Mental Component Summary (MCS) scores, Ankle Osteoarthritis Scale (AOS) pain scale scores, demographic data, and systemic and musculoskeletal comorbidities were determined prospectively for 195 patients with ankle osteoarthritis and ninety-five age and gender-matched controls. The effect of systemic and musculoskeletal comorbidities on each of the scores was determined. RESULTS: On the average, patients with ankle osteoarthritis had a relatively normal MCS score (47 +/- 13 points) and a markedly diminished PCS score (32 +/- 8 points). Both of these scores averaged 50 +/- 9 points in the control group. The AOS pain score averaged 61 +/- 23 points in the group with ankle osteoarthritis, whereas it averaged 10 +/- 15 points in the control group. We found the perception of ankle pain by patients with ankle osteoarthritis to be significantly and linearly associated with the number of other musculoskeletal problems (not related to the foot or ankle). CONCLUSIONS: The degree of physical impairment associated with ankle osteoarthritis, as measured with the SF-36, is equivalent to that reported to be associated with severely disabling medical problems including end-stage kidney disease and congestive heart failure. The perception of ankle health status as measured with a validated, patient-oriented, anatomically specific instrument is influenced by the patients' perception of their overall musculoskeletal comorbidity status. The authors of clinical studies using these instruments should adjust for concomitant musculoskeletal disease. PMID- 17079393 TI - Revision anterior cruciate ligament reconstruction with doubled semitendinosus and gracilis tendons and lateral extra-articular reconstruction. AB - BACKGROUND: The outcome of revision anterior cruciate ligament reconstruction has only rarely been reported. The purpose of this study was to evaluate the results of revision anterior cruciate ligament surgery with use of an autogenous doubled semitendinosus and gracilis graft in association with an extra-articular procedure. METHODS: Between 1997 and 2003, thirty patients underwent a repeat reconstruction of a previously reconstructed torn anterior cruciate ligament with use of a doubled semitendinosus and gracilis graft combined with an extra articular reconstruction. Primary reconstruction had been done with an autogenous patellar tendon graft in twenty-six patients and with a prosthetic ligament in four patients; the average time from the primary reconstruction to the revision was five years. Functional outcomes, graft survival, and radiographic outcomes were evaluated at a mean of five years. A graft was considered to have failed when a revision was done or when the side-to-side difference on KT-1000 arthrometer testing was >5 mm and/or the pivot-shift test grade was greater than a trace. RESULTS: One patient underwent another revision reconstruction because of graft failure at three years postoperatively. The mean International Knee Documentation Committee (IKDC) subjective knee score for the remaining twenty nine patients was 84 +/- 12 points, and the mean Lysholm knee score was 90 +/- 10 points. The side-to-side difference as measured with the KT-1000 arthrometer with maximum manual force was <3 mm in twenty patients (of the twenty-eight who returned for follow-up), between 3 and 5 mm in six patients, and >5 mm in two patients. The result of the pivot shift examination was normal in fifteen patients, slightly positive in eleven patients, and positive in two patients. Twenty-five percent of the patients showed no radiographic signs of degenerative joint disease. CONCLUSIONS: Revision anterior cruciate ligament reconstruction with use of an autogenous doubled semitendinosus and gracilis graft combined with an extra-articular procedure provided satisfactory functional outcomes, with a failure rate of 10%. PMID- 17079394 TI - A comparison of discharge and two-week duplex ultrasound screening protocols for deep venous thrombosis detection following primary total joint arthroplasty. AB - BACKGROUND: Thromboembolic disease in the form of deep venous thrombosis and pulmonary embolism is a risk following hip and knee joint replacement. Prophylactic and screening protocols have been employed to prevent thromboembolic disease following lower extremity joint reconstruction. The purpose of the present study was to evaluate two noninvasive venous screening protocols, specifically, compression ultrasonography performed either at the time of hospital discharge or two weeks after the operation. METHODS: From 1994 through 2001, 2364 patients undergoing primary unilateral total hip or total knee arthroplasty were managed with an anticoagulation chemoprophylaxis protocol (adjusted-dose warfarin) until the time of noninvasive venous screening with use of one of two protocols. Nine hundred thirty-one patients (406 hips and 525 knees) underwent compression ultrasonography prior to hospital discharge, and the other 1433 patients (614 hips and 819 knees) underwent ultrasonographic screening two weeks after the operation. RESULTS: Twenty-three proximal deep venous thromboses (prevalence, 2.5%) were identified in the group that underwent ultrasound screening at the time of hospital discharge, and thirty-one proximal thromboses (prevalence, 2.2%) were identified in the group that underwent ultrasound screening two weeks after the operation. There was no significant difference between the two protocols with regard to the detection of deep venous thrombosis. CONCLUSIONS: There was no significant difference between the group that received two weeks of warfarin chemoprophylactic prophylaxis and the group that was screened at the time of hospital discharge with regard to the detection of deep venous thrombosis with use of compression ultrasound. On the basis of these findings, we no longer screen asymptomatic patients for deep venous thrombosis following hip and knee replacement, and all patients receive warfarin anticoagulation for two weeks. PMID- 17079395 TI - Enhanced fracture and soft-tissue healing by means of anabolic dietary supplementation. AB - BACKGROUND: Malnutrition is common in hospitalized injured patients. It contributes to delayed fracture-healing and increased morbidity. However, relatively little attention has been directed toward nutritional strategies for augmenting musculoskeletal recovery after a fracture. This animal study was designed to examine the effects of dietary protein intake and the role of conditionally essential amino acids in muscle and bone-healing after a fracture. METHODS: One hundred adult male rats were used. Ten rats served as controls and received a 15% protein diet throughout the study. The remaining ninety rats received a 6% protein diet for five weeks to induce protein malnutrition. The rats underwent intramedullary nailing and closed midshaft fracture of one femur. After the fracture, they were separated into three isocaloric dietary groups. Group P6 received a diet with 6% protein; Group P15, a diet with 15% protein; and group P30, a diet with 30% protein with conditionally essential amino acids. At two, four, and six weeks after surgery, ten animals from each group were killed and the femora were evaluated with dual x-ray absorptiometry, histomorphometric assessment of callus, and torsional testing. The quadriceps muscles were analyzed for total mass, total protein content, and for mRNA expression of insulin-like growth factor-1 (IGF-1), IGF-2, IGF receptors, actin, myosin, and vascular endothelial growth factor (VEGF). RESULTS: The P30 group demonstrated elevations in albumin, body mass, muscle mass, total protein content of muscle, and bone mineral density in the fracture callus compared with the P6 diet group at six weeks (p < 0.05). Molecular analysis of muscle revealed that IGF-1, IGF-2, IGF receptors, myosin, actin, and VEGF gene expression were significantly (p < 0.001) higher in the P6 group compared with the P30 group. Biomechanical testing of the femora, however, showed no significant differences. CONCLUSIONS: Dietary supplementation with conditionally essential amino acids in malnourished animals had anabolic effects on bone mineralization, body mass, and muscle mass. PMID- 17079396 TI - Is there a safe area for the axillary nerve in the deltoid muscle? A cadaveric study. AB - BACKGROUND: Several authors have defined a variety of so-called safe zones for deltoid-splitting incisions. The first aim of the present study was to investigate the distance of the axillary nerve from the acromion and its relation to arm length. The second aim was to identify a safe area for the axillary nerve during surgical dissection of the deltoid muscle. METHODS: Twenty-four shoulders of embalmed adult cadavers were included in the study. The distance from the anterior edge of the acromion to the course of the axillary nerve was measured and was recorded as the anterior distance. The same measurement from the posterior edge of the acromion to the course of the axillary nerve was made and was recorded as the posterior distance for each limb. Correlation analysis was performed between the arm length and the anterior distance and the posterior distance for each limb. The ratios between arm length and the anterior and posterior distances were calculated for each case and were recorded as an anterior index and a posterior index. RESULTS: The average arm length was 30.40 cm. The average anterior distance was 6.08 cm, and the average posterior distance was 4.87 cm. There was a significant correlation between arm length and both anterior distance (r = 0.79, p < 0.001) and posterior distance (r = 0.61, p = 0.001). The axillary nerve was not found to lie at a constant distance from the acromion at every point along its course. The average anterior index was 0.20, and the average posterior index was 0.16. CONCLUSIONS: The present study describes a safe area above the axillary nerve that is quadrangular in shape, with the length of the lateral edges being dependent on the individual's arm length. Using this safe area should provide a safe exposure for the axillary nerve during shoulder operations. PMID- 17079397 TI - Transplantation of preconditioned schwann cells in peripheral nerve grafts after contusion in the adult spinal cord. Improvement of recovery in a rat model. AB - BACKGROUND: Recovery after injury to the peripheral nervous system is based on the pro-regenerative relationship between axons and the extracellular matrix, a relationship established by Schwann cells. As mechanical conditioning of Schwann cells has been shown to stimulate their regenerative behavior, we sought to determine whether transplantation of these cells to the central nervous system (i.e., the spinal cord), with its limited regenerative capacity after injury, would improve axonal regeneration and functional recovery. METHODS: A moderate contusion injury of the spinal cord was created with a force-directed impactor in forty-eight adult Sprague-Dawley rats, and, at one week postinjury, the spinal cords were reexposed in all animals. In twenty-four of these animals, peripheral nerve grafts with Schwann cells that had been obtained from the sciatic nerves of donor animals, and had been either untreated or subjected to mechanical conditioning, were transplanted to the contused area of the cords following resection of the glial scar. Another group of animals was treated with glial scar excision only, and a fourth group had the contusion injury but neither glial excision nor transplantation. Scores according to the Basso, Beattie, Bresnahan (BBB) Locomotor Rating Scale were assigned preoperatively and weekly thereafter. Tract tracing of descending and ascending spinal cord tracts was performed at six weeks postoperatively for quantitative histological evaluation of axonal regeneration. RESULTS: While the recovery following glial scar excision without peripheral nerve transplantation was significantly worse than the recovery in the other groups, both transplantation groups had significantly higher BBB scores than the controls (no transplantation) in the early postoperative period (p < 0.05). Moreover, histological analysis showed markedly increased axonal regeneration at the lesional sites in the animals treated with the mechanically conditioned grafts than in the other groups (p < 0.05). CONCLUSIONS: Functional recovery after spinal cord contusion improved following glial scar excision with transplantation of Schwann cells in peripheral nerve grafts to the contusion areas. Although recovery did not differ significantly between the transplantation groups, only the preconditioned grafts led to axonal regeneration at and past the lesional site. These grafts may further enhance functional recovery as the descending tracts eventually reach their target end-organs. PMID- 17079398 TI - Internal fixation of dorsally displaced fractures of the distal part of the radius. A biomechanical analysis of volar plate fracture stability. AB - BACKGROUND: Volar plate fixation with use of either a locking plate or a neutralization plate has become increasingly popular among surgeons for the treatment of dorsally comminuted extra-articular distal radial fractures. The purpose of the present study was to compare the relative stability of five distal radial plates (four volar and one dorsal), all of which are commonly used for the treatment of dorsally comminuted extra-articular distal radial fractures, under loading conditions simulating the physiologic forces that are experienced during early active rehabilitation. METHODS: With use of a previously validated Sawbones fracture model, a dorsally comminuted extra-articular distal radial fracture was created. The fracture fixation stability of four volar plates (an AO T-plate, an AO 3.5-mm small-fragment plate, an AO 3.5-mm small-fragment locking plate, and the Hand Innovations DVR locking plate) were compared under axial compression loading and dorsal and volar bending simulating the in vivo stresses that are generated at the fracture site during early unopposed active motion of the wrist and digits. A single dorsal plate (an AO pi plate) was used for comparison, with and without simulated volar cortical comminution. The construct stiffness was measured to assess the resistance to fracture gap motion, and comparisons were made among the implants. RESULTS: The volar AO locking and DVR plates had greater resistance to fracture gap motion (greater stiffness) compared with the volar AO nonlocking and AO T-plates under axial and dorsal loading conditions (p < 0.01), with no significant difference between the AO volar locking and DVR plates. The volar AO locking plate had greater resistance to fracture gap motion than did the volar AO nonlocking plate under axial loading and dorsal bending forces (p < 0.01). The dorsal pi plate had the greatest resistance to fracture gap motion under axial loading and volar and dorsal bending forces (p < 0.01). However, the pi plate was significantly less stable to axial load and dorsal bending forces when the volar cortex was comminuted (p < 0.01). CONCLUSIONS: In this model of dorsally comminuted extra-articular distal radial fractures, dorsal pi-plate fixation demonstrated better resistance to fracture gap motion than did the four types of volar plate fixation. The AO volar locking and DVR plates conferred the greatest resistance to fracture gap motion among the four volar plates tested. Volar locking technology conferred a significant increase in resistance to fracture gap motion as compared with nonlocking plate technology. PMID- 17079399 TI - The extruded talus: results of reimplantation. AB - BACKGROUND: There is considerable debate regarding the appropriate treatment of the extruded talus regardless of the presence of a fracture. The purpose of this study was to report the clinical results, complications, and functional outcome following reimplantation of the traumatically extruded talus. METHODS: A database of 119 patients with an open injury of the talus occurring between 1995 and 2003 at a level-I trauma center was reviewed to identify patients with a complete talar extrusion. Demographic, imaging, and treatment data were obtained from a review of the medical records. Follow-up was undertaken during clinic visits or by telephone. Preoperative and follow-up radiographs were reviewed to identify posttraumatic arthritis, osteonecrosis, or talar collapse, and the Musculoskeletal Functional Assessment was used to assess functional outcome. RESULTS: Twenty-seven patients were identified. A minimum follow-up of one year (average, forty-two months) was obtained for nineteen patients. Infection and the need for a secondary surgical procedure were the primary determinants of clinical outcome. Two of the nineteen patients had documented infections: one had developed at two weeks and one, after a calcaneal osteotomy at nineteen months. Twelve patients had no subsequent surgery, and seven had subsequent procedures (range, one to four procedures). No patient underwent a delayed amputation. The average Musculoskeletal Functional Assessment score at the time of follow-up was 29.8 (range, 5 to 59). With the numbers studied, no association was found between functional outcome and the following variables: ipsilateral lower-extremity injury, associated talar fracture, secondary procedures, osteonecrosis, or age. CONCLUSIONS: While functional outcome is difficult to assess, salvage of the extruded talus appears to be a relatively safe operation, with a minimal risk of infection, which allows maximal flexibility in aftercare by preserving the most normal ankle anatomy possible. PMID- 17079400 TI - A biomechanical comparison of single and double-row fixation in arthroscopic rotator cuff repair. AB - BACKGROUND: The optimal method for arthroscopic rotator cuff repair is not yet known. The hypothesis of the present study was that a double-row repair would demonstrate superior static and cyclic mechanical behavior when compared with a single-row repair. The specific aims were to measure gap formation at the bone tendon interface under static creep loading and the ultimate strength and mode of failure of both methods of repair under cyclic loading. METHODS: A standardized tear of the supraspinatus tendon was created in sixteen fresh cadaveric shoulders. Arthroscopic rotator cuff repairs were performed with use of either a double-row technique (eight specimens) or a single-row technique (eight specimens) with nonabsorbable sutures that were double-loaded on a titanium suture anchor. The repairs were loaded statically for one hour, and the gap formation was measured. Cyclic loading to failure was then performed. RESULTS: Gap formation during static loading was significantly greater in the single-row group than in the double-row group (mean and standard deviation, 5.0 +/- 1.2 mm compared with 3.8 +/- 1.4 mm; p < 0.05). Under cyclic loading, the double-row repairs failed at a mean of 320 +/- 96.9 N whereas the single-row repairs failed at a mean of 224 +/- 147.9 N (p = 0.058). Three single-row repairs and three double-row repairs failed as a result of suture cut-through. Four single-row repairs and one double-row repair failed as a result of anchor or suture failure. The remaining five repairs did not fail, and a midsubstance tear of the tendon occurred. CONCLUSIONS: Although more technically demanding, the double-row technique demonstrates superior resistance to gap formation under static loading as compared with the single-row technique. CLINICAL RELEVANCE: A double-row reconstruction of the supraspinatus tendon insertion may provide a more reliable construct than a single-row repair and could be used as an alternative to open reconstruction for the treatment of isolated tears. PMID- 17079401 TI - Preservation of the ulnar bursa within the carpal tunnel: does it improve the outcome of carpal tunnel surgery? A randomized, controlled trial. AB - BACKGROUND: It was hypothesized that preserving a layer of gliding tissue, the parietal layer of the ulnar bursa, between the contents of the carpal tunnel and the soft tissues incised during carpal tunnel surgery might reduce scar pain and improve grip strength and function following open carpal tunnel decompression. METHODS: Patients consented to randomization to treatment with either preservation of the parietal layer of the ulnar bursa beneath the flexor retinaculum at the time of open carpal tunnel decompression (fifty-seven patients) or division of this gliding layer as part of a standard open carpal tunnel decompression (sixty-one patients). Grip strength was measured, scar pain was rated, and the validated Patient Evaluation Measure questionnaire was used to assess symptoms and disability preoperatively and at eight to nine weeks following the surgery in seventy-seven women and thirty-four men; the remaining seven patients were lost to follow-up. RESULTS: There was no difference between the groups with respect to age, sex, hand dominance, or side of surgery. Grip strength, scar pain, and the Patient Evaluation Measure score were not significantly different between the two groups, although there was a trend toward a poorer subjective outcome as demonstrated by the questionnaire in the group in which the ulnar bursa within the carpal tunnel had been preserved. Preserving the ulnar bursa within the carpal tunnel did, however, result in a lower prevalence of suspected wound infection or inflammation (p = 0.04). CONCLUSIONS: In this group of patients, preservation of the ulnar bursa around the median nerve during open carpal tunnel release produced no significant difference in grip strength or self-rated symptoms. We recommend incision of the ulnar bursa during open carpal tunnel decompression to allow complete visualization of the median nerve and carpal tunnel contents. PMID- 17079402 TI - Influence of lower-limb torsion on long-term outcomes of tibial valgus osteotomy for medial compartment knee osteoarthritis. AB - BACKGROUND: The results of tibial osteotomy used to treat osteoarthritis of the medial compartment of the knee deteriorate over time even when the initial correction is optimal. Studies have shown that tibial and femoral torsion and the femorotibial index (tibial torsion minus femoral torsion) contribute, together with coronal malalignment, to the development of single-compartment knee osteoarthritis. The objective of our study was to evaluate the impact of femoral and tibial torsion and of coronal realignment on the long-term clinical and radiographic outcomes of valgus tibial osteotomy. METHODS: A function score was calculated for sixty-eight patients at a mean of thirteen years after the osteotomy. Anteroposterior single-leg-stance radiographs were used to evaluate loss of the femorotibial joint space. Goniometry was used to measure coronal malalignment preoperatively, at one year, and at the time of the last follow-up, and postoperative computed tomography was performed to measure femoral anteversion and tibial torsion and to calculate the femorotibial index. We looked for associations linking body mass index, initial loss of joint space, coronal malalignment, femoral and tibial torsion, the femorotibial index, and functional outcomes. RESULTS: Worse outcomes were associated with changes in coronal alignment (>/=2 degrees ) over time, which were associated with deterioration of the femorotibial space. Femoral anteversion was significantly greater in patients in whom valgus increased over time than in those in whom valgus decreased over time. Stability of coronal alignment seemed to be dependent on a linear relationship between the femorotibial index and the degree of postoperative realignment. A body mass index of >25 kg/m(2) was associated with a long-term loss of coronal realignment. Preoperative loss of the medial femorotibial joint space, coronal alignment at one year, and age were not associated with secondary malalignment or functional outcomes. CONCLUSIONS: Long-term success of a valgus tibial osteotomy is related to the stability over time of the postoperative coronal realignment. Therefore, the results of our study suggest that modifying the realignment according to the extent of femoral anteversion may improve long term outcomes. PMID- 17079403 TI - Arthroscopic treatment of partial scapholunate ligament tears in children with chronic wrist pain. AB - BACKGROUND: Scapholunate ligament injury is rare in the adolescent and pediatric population, and the results of treatment have not been well described. The purpose of the present study was to review the outcomes of arthroscopic management of patients with persistent wrist pain and scapholunate ligament injury as documented on arthroscopic examination who had had a failure of at least six months of nonoperative management. METHODS: The medical records of thirty-two pediatric and adolescent patients who underwent arthroscopic treatment of scapholunate ligament injuries were retrospectively reviewed to obtain preoperative and postoperative modified Mayo wrist scores, radiographic data, and intraoperative findings, including the classifications of interosseous ligament injury, chondral injury, and other abnormalities. Patients were contacted after a minimum of two years of follow-up for reevaluation of the Mayo wrist scores. RESULTS: Arthroscopic evaluation revealed thirty Geissler type-II tears and two Geissler type-III tears. In addition to these scapholunate ligament injuries, seven of the thirty-two patients had partial tears of the short radiolunate ligament that appeared to be at the site of impaction from carpal subluxation, twelve had a triangular fibrocartilage complex injury, and twenty-seven had a chondral injury. The modified Mayo wrist scores showed improvement following arthroscopic debridement of partial-thickness tears and associated chondral injuries. The average wrist score was 66.3 preoperatively and 91.6 at an average of forty-three months of follow-up. Eight patients required subsequent surgery because of deterioration in their clinical status. After a mean duration of follow-up of 30.8 months, the average wrist score was 87.1. CONCLUSIONS: The majority of pediatric and adolescent patients with wrist pain who have a failure of nonoperative management and who have a Geissler type-II scapholunate ligament tear on arthroscopic examination can have substantial long-term improvement following arthroscopic debridement of the tear combined with treatment of other associated injuries. PMID- 17079404 TI - Operative treatment of primary synovial osteochondromatosis of the hip. AB - BACKGROUND: Primary synovial osteochondromatosis of the hip, a rare benign condition characterized by multiple intra-articular osteochondral loose bodies and synovial hyperplasia, may result in mechanical symptoms and degenerative arthritis if untreated. The purpose of this study was to report the results of arthrotomy alone or combined with anterior dislocation of the hip to perform synovectomy and removal of loose bodies in patients with this condition. METHODS: We retrospectively reviewed a consecutive series of twenty-one patients (twenty one hips) with primary synovial osteochondromatosis of the hip treated with open surgical debridement. On the basis of the extent of extra-articular involvement as seen on preoperative magnetic resonance imaging, eight of the twenty-one patients underwent synovectomy and removal of loose bodies following anterior dislocation of the hip and thirteen underwent the same procedure with arthrotomy alone. At a mean of 4.4 years postoperatively, the patients were assessed clinically and radiographically with special attention to disease recurrence, osteoarthritis progression, and surgical complications. RESULTS: The mean Harris hip score for the entire series of patients improved from 58 points preoperatively to 91 points at the time of the latest follow-up. Eighteen of the twenty-one patients had a good or excellent clinical result, and seventeen patients were satisfied with the result of the surgery. The clinical scores, patient satisfaction scores, and radiographic grades of osteoarthritis at the time of the latest follow-up did not differ significantly between the group treated with dislocation and the group treated without dislocation. Symptomatic disease recurred in two of the thirteen hips treated with arthrotomy alone and in none of the hips that had undergone dislocation. However, the surgical complication rate was higher in the group treated with dislocation than it was in the group treated without dislocation (p = 0.042). While patients with some signs of mild osteoarthritis at the initial procedure had a higher rate of osteoarthritis progression, severe osteoarthritis requiring arthroplasty had developed in only one patient at the time of follow-up. CONCLUSIONS: At a mean of 4.4 years postoperatively, we found that open synovectomy and removal of loose bodies for the treatment of primary synovial osteochondromatosis of the hip is a reliable procedure that can effectively relieve symptoms. Our results also indicated that synovial osteochondromatosis may recur in patients with extensive involvement who are treated with synovectomy alone without dislocation of the hip; however, surgical complications are more likely to occur in patients managed with anterior dislocation of the hip and synovectomy. PMID- 17079405 TI - Mannose-6-phosphate, an inhibitor of transforming growth factor-beta, improves range of motion after flexor tendon repair. AB - BACKGROUND: Adhesion formation between the flexor tendon and its surrounding fibro-osseous sheath results in a decreased postoperative range of motion in the hand. Transforming growth factor-beta (TGF-beta) is a key cytokine in the pathogenesis of tissue fibrosis. In this study, the effects of two natural inhibitors of TGF-beta, decorin and mannose-6-phosphate, were investigated in vitro and in vivo. METHODS: In the in vitro investigation, primary cell cultures from rabbit flexor tendon sheath, epitenon, and endotenon were established and each was supplemented with TGF-beta along with increasing doses of decorin or mannose-6-phosphate. Collagen-I production was measured with enzyme-linked immunosorbent assay (ELISA). For the in vivo study, rabbit zone-II flexor tendons were transected and then immediately repaired. Single intraoperative graded doses of decorin, mannose-6-phosphate, or phosphate-buffered saline solution (control) were added to the repair sites, and the forepaws were tested for the range of motion and repair strength at eight weeks postoperatively. RESULTS: Decorin and mannose-6-phosphate both reduced TGF-beta upregulated collagen production. Intraoperative application of low-dose mannose-6-phosphate significantly improved the range of motion of the operatively treated digits. The effect on breaking strength of the tendon repair was inconclusive. CONCLUSIONS: Mannose-6-phosphate is effective in reducing TGF-beta upregulated collagen production in an in vitro model. This finding correlated with our in vivo finding that a single intraoperative dose of mannose-6-phosphate improved the postoperative range of motion. PMID- 17079406 TI - Thigh mass and lytic diaphyseal femoral lesion associated with polyethylene wear after hybrid total knee arthroplasty. A case report. PMID- 17079407 TI - Cervical spine subluxation in Marfan syndrome. A case report. PMID- 17079408 TI - Vascular injury during fixation of an intertrochanteric hip fracture in a patient with severe atherosclerosis. A case report. PMID- 17079410 TI - Orthopaedic gifts: opportunities and obligations. PMID- 17079409 TI - Antibiotic-loaded bone cement for infection prophylaxis in total joint replacement. AB - Use of antibiotic-loaded bone cement for prophylaxis against infection is not indicated for patients not at high risk for infection who are undergoing routine primary or revision joint replacement with cement. The mechanical and elution properties of commercially available premixed antibiotic-loaded bone-cement products are superior to those of hand-mixed preparations. Use of commercially available antibiotic-loaded bone-cement products has been cleared by the United States Food and Drug Administration only for use in the second stage of a two stage total joint revision following removal of the original prosthesis and elimination of active periprosthetic infection. Use of antibiotic-loaded bone cement for prophylaxis against infection in the second stage of a two-stage total joint revision involves low doses of antibiotics. Active infection cannot be treated with commercially available antibiotic-loaded bone cement as such treatment requires higher doses of antibiotics. PMID- 17079411 TI - Topics in medical economics: health care rationing. PMID- 17079413 TI - Concern regarding duplicate publications. PMID- 17079414 TI - Treatment of external fixation pins about the wrist. PMID- 17079415 TI - Evaluation of a single-dose, extended-release epidural morphine formulation for pain after knee arthroplasty. PMID- 17079417 TI - Long-term follow-up of patients with clubfeet treated with extensive soft-tissue release. PMID- 17079420 TI - Treatment of femoro-acetabular impingement: preliminary results of labral refixation. PMID- 17079422 TI - Autologous cultured chondrocytes: adverse events reports. PMID- 17079423 TI - Autologous cultured chondrocytes: adverse events reports. PMID- 17079424 TI - Failure of LCP condylar plate fixation in the distal part of the femur. PMID- 17079429 TI - Coping with racism in a patient. PMID- 17079430 TI - What's new in orthopaedic trauma. PMID- 17079431 TI - Specificity of the immune response to a modified group B meningococcal polysaccharide conjugate vaccine. AB - Antibodies to a modified group B meningococcal polysaccharide vaccine were examined for antigenic and functional specificities. Bactericidal determinants were investigated by using immunoaffinity columns and competitive inhibition of bactericidal activity in an in vitro killing assay. We conclude that nearly all of the vaccine-induced bactericidal activity is specific for the native polysaccharide. PMID- 17079432 TI - Rapid expression of Mycobacterium avium subsp. paratuberculosis recombinant proteins for antigen discovery. AB - Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne's disease, a chronic granulomatous enteritis of ruminants and other species. Detection of infection in animals is hampered by the lack of sensitive and specific diagnostic assays. We describe here an approach that utilizes translationally active PCR fragments for the rapid in vitro transcription and translation of recombinant proteins for antigen discovery in M. avium subsp. paratuberculosis. The investigations showed that the MAP1272c protein selectively reacts with sera from Johne's disease-positive cattle and represents an antigen of potential utility in M. avium subsp. paratuberculosis immunodiagnostics. PMID- 17079433 TI - Dynamics of Helicobacter pylori-specific immunoglobulin G for 2 years after successful eradication of Helicobacter pylori infection in an American Indian and Alaska Native population. AB - Helicobacter pylori antibodies were measured over 24 months in American Indian and Alaska Native persons who cleared their infections. Two months after treatment, 82% of H. pylori-negative persons remained seropositive. While there were declines in H. pylori antibodies for 12 months, after 24 months 71% of persons remained seropositive. PMID- 17079434 TI - Test for detection of disease-associated prion aggregate in the blood of infected but asymptomatic animals. AB - We have developed a sensitive in vitro assay for detecting disease-associated prion aggregates by combining an aggregation-specific enzyme-linked immunosorbent assay (AS-ELISA) with the fluorescent amplification catalyzed by T7 RNA polymerase technique (FACTT). The new assay, named aggregation-specific FACTT (AS FACTT), is much more sensitive than AS-ELISA and could detect prion aggregates in the brain of mice as early as 7 days after an intraperitoneal inoculation of PrP(Sc). However, AS-FACTT was still unable to detect prion aggregates in blood of infected mice. To further improve the detection limit of AS-FACTT, we added an additional prion amplification step (Am) and developed a third-generation assay, termed Am-A-FACTT. Am-A-FACTT has 100% sensitivity and specificity in detecting disease-associated prion aggregates in blood of infected mice at late but still asymptomatic stages of disease. At a very early stage, Am-A-FACTT had a sensitivity of 50% and a specificity of 100%. Most importantly, Am-A-FACTT also detects prion aggregates in blood of mule deer infected with the agent causing a naturally occurring prion disease, chronic wasting disease. Application of this assay to cattle, sheep, and humans could safeguard food supplies and prevent human contagion. PMID- 17079435 TI - Human prolactin promotes human secondary immunoglobulin response in human/SCID mouse chimeras. AB - Recombinant human prolactin (rhPRL) was administered to huPBL-SCID mice to determine its effects on production of human immunoglobulin (Ig). The huPBL-SCID mice were injected intraperitoneally (i.p.) with 10 mug rhPRL every other day for a total of 10 injections. The results reconfirmed that rhPRL significantly increased the numbers of human CD3+ T cells and human CD19+ B cells in spleens, lymph nodes, and thymuses of huPBL-SCID mice. The huPBL-SCID mice were then concurrently given various doses of diphtheria-tetanus (DT) vaccine and 10-mug i.p. injections of rhPRL and were examined for the presence of human DT-specific proliferation of lymph node cells in vitro and antibody production in vivo. rhPRL greatly improved the engraftment of functional human lymphocytes (CD3+ T cells and CD19+ B cells) in DT-immunized huPBL-SCID mice. The rhPRL-treated, DT immunized huPBL-SCID mice produced significantly larger amounts of DT-specific antibodies in response to the vaccine. The predominant Ig isotype induced after immunization was IgG. Thus, rhPRL stimulation promotes human secondary IgG responses in huPBL-SCID mice. PMID- 17079436 TI - CD27 and CD57 expression reveals atypical differentiation of human immunodeficiency virus type 1-specific memory CD8+ T cells. AB - The failure of human immunodeficiency virus type 1 (HIV-1)-specific CD8+ T cells to control chronic HIV-1 infection could be due to the progressive loss of their capacities to undergo normal memory effector differentiation. We characterized and compared the expressions of CD27, CD28, CD57, and CD62L by Epstein-Barr virus (EBV)-, cytomegalovirus (CMV)-, and HIV-1-specific CD8+ T cells by six-color, eight-parameter flow cytometry. In contrast to the maturation of EBV- and CMV specific memory CD8+ T cells, we found that HIV-1-specific CD8+ T cells did not display coordinated down-regulation of CD27 and up-regulation of CD57 and accumulated in an atypical CD27(high) CD57(low) subset. Moreover, the accumulation of CD27(high) CD57(low) HIV-1-specific CD8+ T cells was positively correlated with HIV-1 plasma viremia. The differentiation of HIV-1-specific CD8+ T cells to an effector subset is therefore impaired during chronic HIV-1 infection. This lack of normal CD8+ T-cell differentiation could contribute to the failure of cellular immune control of HIV-1 infection. PMID- 17079437 TI - Prospects for cervical cancer prevention by human papillomavirus vaccination. AB - Recent clinical trials in young women have shown that subunit vaccines based on human papillomavirus (HPV) 16 and HPV18 L1 virus-like particles are approximately 100% effective in short-term prevention of persistent cervical infection and of cervical dysplasia by these major oncogenic types. These remarkable efficacy results, together with an excellent safety profile in thousands of vaccinated women, have led to the HPV prophylactic vaccine from one manufacturer having now been licensed for commercial use and the expectation that the vaccine from a second manufacturer will be approved in the near future. These vaccines seem to have great potential for reducing cervical cancer deaths and treatments to remove premalignant cervical lesions. However, before their public health effect can be fully estimated, several issues must be addressed. These include duration of protection, degree of cross-protection against nonvaccine types, efficacy in men, and vaccine availability to economically disadvantaged women. PMID- 17079438 TI - Heparanase, hyaluronan, and CD44 in cancers: a breast carcinoma perspective. AB - Glycosaminoglycans are major constituents of the cancer cell surface and the tumor stroma. The heparan sulfate degrading enzyme heparanase, hyaluronan, and its receptor CD44 are up-regulated in breast cancer, generating a microenvironment that promotes tumor progression and metastasis. Recent experimental and clinical evidence shows that heparanase, hyaluronan, and CD44 regulate cancer cell proliferation, migration, and invasion, as well as tumor associated angiogenesis and are correlated with patient survival. These findings suggest that they may be used as prognostic factors and targets for breast cancer treatment. PMID- 17079439 TI - Lysyl oxidase mediates hypoxic control of metastasis. AB - Hypoxic cancer cells pose a great challenge to the oncologist because they are especially aggressive, metastatic, and resistant to therapy. Recently, we showed that elevation of the extracellular matrix protein lysyl oxidase (LOX) correlates with metastatic disease and is essential for hypoxia-induced metastasis. In an orthotopic rodent model of breast cancer, a small-molecule or antibody inhibitor of LOX abolished metastasis, offering preclinical validation of this enzyme as a therapeutic target. PMID- 17079440 TI - TMPRSS2 fusions with oncogenic ETS factors in prostate cancer involve unbalanced genomic rearrangements and are associated with HDAC1 and epigenetic reprogramming. AB - Translocations fusing the strong androgen-responsive gene, TMPRSS2, with ERG or other oncogenic ETS factors may facilitate prostate cancer development. Here, we studied 18 advanced prostate cancers for ETS factor alterations, using reverse transcription-PCR and DNA and RNA array technologies, and identified putative ERG downstream gene targets from the microarray data of 410 prostate samples. Out of the 27 ETS factors, ERG was most frequently overexpressed. Seven cases showed TMPRSS2:ERG gene fusions, whereas the TMPRSS2:ETV4 fusion was seen in one case. In five out of six tumors with high ERG expression, array-CGH analysis revealed interstitial 2.8 Mb deletions between the TMPRSS2 and ERG loci, or smaller, unbalanced rearrangements. In silico analysis of the ERG gene coexpression patterns revealed an association with high expression of the histone deacetylase 1 gene, and low expression of its target genes. Furthermore, we observed increased expression of WNT-associated pathways and down-regulation of tumor necrosis factor and cell death pathways. In summary, our data indicate that the TMPRSS2:ERG translocation is common in advanced prostate cancer and occurs by virtue of unbalanced genomic rearrangements. Activation of ERG by fusion with TMPRSS2 may lead to epigenetic reprogramming, WNT signaling, and down-regulation of cell death pathways, implicating ERG in several hallmarks of cancer with potential therapeutic importance. PMID- 17079441 TI - Neurospheres enriched in cancer stem-like cells are highly effective in eliciting a dendritic cell-mediated immune response against malignant gliomas. AB - Cancer stem-like cells (CSC) could be a novel target for cancer therapy, including dendritic cell (DC) immunotherapy. To address this, we developed experiments aimed at DC targeting of neurospheres (NS) from GL261 glioma cells because neurospheres can be enriched in CSC. We obtained murine neurospheres by growing GL261 cells in epidermal growth factor/basic fibroblast growth factor without serum. GL261-NS recapitulated important features of glioblastoma CSC and expressed higher levels of radial glia stem cell markers than GL261 cells growing under standard conditions (GL261 adherent cells, GL261-AC), as assessed by DNA microarray and real-time PCR. GL261-NS brain gliomas were highly infiltrating and more rapidly lethal than GL261-AC, as evidenced by survival analysis (P < 0.0001), magnetic resonance imaging and histology. DC from the bone marrow of syngeneic mice were then used for immunotherapy of GL261-NS and GL261-AC tumors. Strikingly, DC loaded with GL261-NS (DC-NS) cured 80% and 60% of GL261-AC and GL261-NS tumors, respectively (P < 0.0001), whereas DC-AC cured only 50% of GL261 AC tumors (P = 0.0022) and none of the GL261-NS tumors. GL261-NS expressed higher levels of MHC and costimulatory molecules (CD80 and CD86) than GL261-AC; the JAM assay indicated that DC-NS splenocytes had higher lytic activity than DC-AC splenocytes on both GL261-NS and GL261-AC, and immunohistochemistry showed that DC-NS vaccination was associated with robust tumor infiltration by CD8+ and CD4+ T lymphocytes. These findings suggest that DC targeting of CSC provides a higher level of protection against GL261 gliomas, a finding with potential implications for the design of clinical trials based on DC vaccination. PMID- 17079442 TI - Adaptation to the ionizing radiation-induced G2 checkpoint occurs in human cells and depends on checkpoint kinase 1 and Polo-like kinase 1 kinases. AB - Checkpoint adaptation was originally defined in yeast as the ability to divide despite the presence of damaged DNA. An important unanswered question is whether checkpoint adaptation also occurs in human cells. Here, we show that following the ionizing radiation-induced G(2) checkpoint, human osteosarcoma cells entered mitosis with gamma-H2AX foci, a marker for unrepaired DNA double-strand breaks. Exit from the G(2) checkpoint was accelerated by inhibiting the checkpoint kinase 1 (Chk1) and delayed by overexpressing wild-type Chk1 or depleting the Polo-like kinase 1 (Plk1). Chk1 and Plk1 controlled this process, at least partly, via independent signaling pathways. Our results suggest that human cells are able to exit the checkpoint arrest and divide before the damage has been fully repaired. Such cell division in the presence of damaged DNA may be detrimental for genetic stability and could potentially contribute to cancer development. PMID- 17079443 TI - Retinoblastoma pathway defects show differential ability to activate the constitutive DNA damage response in human tumorigenesis. AB - Loss of G(1)-S control and aberrations of the p16(Ink4a)-cyclin D1/cyclin dependent kinase (CDK) 4(6)-pRb-E2F-cyclin E/CDK2 pathway are common in human cancer. Previous studies showed that oncogene-induced aberrant proliferation, such as on cyclin E overexpression, causes DNA damage and checkpoint activation. Here, we show that, in a series of human colorectal adenomas, those with deregulation of cyclin D1 and/or p16(Ink4a) showed little evidence of constitutive DNA damage response (DDR), contrary to cyclin E-overexpressing higher-grade cases. These observations were consistent with diverse cell culture models with differential defects of retinoblastoma pathway components, as overexpression of cyclin D1 or lack of p16(Ink4a), either alone or combined, did not elicit detectable DDR. In contrast, inactivation of pRb, the key component of the pathway, activated the DDR in cultured human or mouse cells, analogous to elevated cyclin E. These results highlight differential effect of diverse oncogenic events on driving the 'cancer cell cycles' and their ability to deregulate the replication-driving CDK2 kinase and to alarm the DDR as a potential anticancer barrier in accordance with their hierarchical positions along the retinoblastoma pathway. Our data provide new insights into oncogene evoked DDR in human tumorigenesis, with potential implications for individualized management of tumors with elevated cyclin D1 versus cyclin E, due to their distinct clinical variables and biological behavior. PMID- 17079444 TI - Expression and regulatory role of GAIP-interacting protein GIPC in pancreatic adenocarcinoma. AB - Regulator of G-protein signaling-GAIP-interacting protein COOH terminus (GIPC) is involved in protein trafficking, endocytosis, and receptor clustering and is associated with insulin-like growth factor I receptor (IGF-IR), a receptor important for proliferation and anchorage-independent growth. Here, we described GIPC expression in different human pancreatic adenocarcinoma (PCA) cell lines and we examined the role of GIPC in the regulation of IGF-IR protein levels in PCA. Interestingly, inhibition of GIPC expression by RNA interference led to reduced IGF-IR protein levels and a subsequent decrease in proliferation of PCA cells. We also determined that the PDZ domain of GIPC is essential for the post translational regulation and the binding of IGF-IR. The importance of GIPC in pancreatic cancer development and progression is supported by tissue microarray data of 300 pancreatic cancer specimens where GIPC is highly expressed in PCA. Taken together, our data suggest that GIPC is a central molecule for the stability of IGF-IR and could be a target for future therapy. PMID- 17079445 TI - Nongenotoxic p53 activation protects cells against S-phase-specific chemotherapy. AB - Mutations in the tumor suppressor gene TP53 represent the most frequent genetic difference between tumor cells and normal cells. Here, we have attempted to turn this difference into an advantage for normal cells during therapy. Using the Mdm2 antagonist nutlin-3, we first activated p53 in U2OS and HCT116 cells to induce cell cycle arrest. These arrested cells were found to be resistant to subsequent transient treatment with the nucleoside analogue gemcitabine, as revealed by clonogenic assays following drug removal. In contrast, isogenic cells lacking functional p53 continued to enter S phase regardless of nutlin-3 pretreatment and remained highly susceptible to gemcitabine-mediated cytotoxicity. The sequential treatment with nutlin-3 alone, followed by transient exposure to nutlin-3 plus gemcitabine, efficiently compromised the clonogenicity of tumor cells with deletions or mutations of p53 but largely spared the proliferation of nontransformed human keratinocytes. Nutlin-3 pretreatment also conferred protection of p53-proficient cells against cytosine arabinoside but not against doxorubicin or cisplatin. We propose that the cell cycle arrest function of p53 can be used to convert p53 from a killer to a protector of cells, with the potential to reduce unwanted side effects of chemotherapy. PMID- 17079446 TI - Aromatase destabilizer: novel action of exemestane, a food and drug administration-approved aromatase inhibitor. AB - Using Western blot as the major technique, we studied the effects of the three Food and Drug Administration (FDA)-approved aromatase inhibitors (AI) on aromatase protein stability in the aromatase-overexpressing breast cancer cell line MCF-7aro. We have found that exemestane treatment significantly reduces aromatase protein level. Exemestane induces aromatase degradation in a dose responsive manner (25-200 nmol/L), and the effect can be seen in as early as 2 hours. Metabolic labeling with S(35)-methionine was used to determine the half life (t(1/2)) of aromatase protein. In the presence of 200 nmol/L exemestane, the t(1/2) of aromatase was reduced to 12.5 hours from 28.2 hours in the untreated cells. Furthermore, exemestane-induced aromatase degradation can be completely blocked by 10 micromol/L MG132, indicating that the degradation is mediated by proteasome. We also examined the effect of exemestane on aromatase mRNA level using real-time reverse transcription-PCR. No significant changes in mRNA level were detected after 8 hours of treatment with exemestane (200 nmol/L). This is the first report on the evaluation of three FDA-approved AIs on the stability of the aromatase protein. We have found that exemestane, different from letrozole and anastrozole, can destabilize the aromatase protein. PMID- 17079447 TI - Alterations of the tumor suppressor gene ARLTS1 in ovarian cancer. AB - ARLTS1 is a tumor suppressor gene initially described as a low-penetrance cancer gene: a truncated Trp149Stop (MUT) polymorphism is associated with general familial cancer aggregation and, particularly, high-risk familial breast cancer. DNA hypermethylation has been identified as a mechanism of ARLTS1 expression down regulation in lung carcinomas and B-cell chronic lymphocytic leukemia. We found that, in the majority of ovarian carcinomas (61.5%) and in a significant proportion of ovarian and breast cancer cell lines (45%), ARLTS1 is strongly down regulated due to DNA methylation in its promoter region. After ARLTS1 restoration by adenoviral transduction, only the negative TOV-112 and the homozygously mutated (MUT) MCF7 cells, but not the OV-90 cells expressing a normal ARLTS1 product, underwent apoptosis and inhibition of cell growth. Furthermore, ARLTS1 reexpression significantly reduced the tumorigenic potential of TOV-112 in nude mice. On the contrary, the ARLTS1-MUT induced significantly lower levels of apoptosis in infected cells and reduced in vivo tumorigenesis only partially, supporting the hypothesis that Trp149Stop polymorphism is retained in the general population and predisposes to cancer because of a reduction, but not full loss, of normal ARLTS1 function. PMID- 17079448 TI - Genetic reclassification of histologic grade delineates new clinical subtypes of breast cancer. AB - Histologic grading of breast cancer defines morphologic subtypes informative of metastatic potential, although not without considerable interobserver disagreement and clinical heterogeneity particularly among the moderately differentiated grade 2 (G2) tumors. We posited that a gene expression signature capable of discerning tumors of grade 1 (G1) and grade 3 (G3) histology might provide a more objective measure of grade with prognostic benefit for patients with G2 disease. To this end, we studied the expression profiles of 347 primary invasive breast tumors analyzed on Affymetrix microarrays. Using class prediction algorithms, we identified 264 robust grade-associated markers, six of which could accurately classify G1 and G3 tumors, and separate G2 tumors into two highly discriminant classes (termed G2a and G2b genetic grades) with patient survival outcomes highly similar to those with G1 and G3 histology, respectively. Statistical analysis of conventional clinical variables further distinguished G2a and G2b subtypes from each other, but also from histologic G1 and G3 tumors. In multivariate analyses, genetic grade was consistently found to be an independent prognostic indicator of disease recurrence comparable with that of lymph node status and tumor size. When incorporated into the Nottingham prognostic index, genetic grade enhanced detection of patients with less harmful tumors, likely to benefit little from adjuvant therapy. Our findings show that a genetic grade signature can improve prognosis and therapeutic planning for breast cancer patients, and support the view that low- and high-grade disease, as defined genetically, reflect independent pathobiological entities rather than a continuum of cancer progression. PMID- 17079449 TI - Truncating variants in p53AIP1 disrupting DNA damage-induced apoptosis are associated with prostate cancer risk. AB - Germ line mutations in several genes (BRCA1, BRCA2, and CHEK2) whose products are involved in the DNA damage-signaling pathway have been implicated in prostate cancer risk. To identify additional genes in this pathway that might confer susceptibility to this cancer, we analyzed a recently identified DNA damage response gene, p53AIP1 (a gene encoding for p53-regulated apoptosis-inducing protein 1), for genetic variants in prostate cancer. Five novel germ line variants were identified. The two truncating variants (Ser(32)Stop and Arg(21)insG) were found in 3% (4 of 132) of unselected prostate tumor samples. Genotyping of the two variants in an additional 393 men with sporadic prostate cancer showed a frequency of 3.1% (12 of 393) in contrast to 0.6% (2 of 327) in 327 unaffected men (Fisher's exact test, P = 0.018), with an odds ratio (OR) of 5.1 [95% confidence interval (95% CI), 1.1-23.0]. In addition, two of six tumors carrying the truncating variants were associated with loss of heterozygosity of the wild-type alleles, suggesting that p53AIP1 may act as a tumor suppressor. We also showed that the truncated p53AIP1 was unable to induce apoptosis and suppress cell growth in HeLa and COS-7 cells. These results suggest that loss-of function variants in p53AIP1 associated with the risk of sporadic prostate cancer and further support the concept that the genetic defects in the DNA damage response genes play an important role in the development of prostate cancer. PMID- 17079450 TI - A repressive epigenetic domino effect confers susceptibility to breast epithelial cell transformation: implications for predicting breast cancer risk. AB - Retinoic acid (RA) is a master epigenetic regulator that plays a pivotal role in both breast morphogenesis and development. Here, we show for the first time that RA, via the RA receptor alpha (RARalpha), epigenetically regulates in a concerted fashion the transcription of two RA-responsive genes, the RA receptor beta2 (RARbeta2) and the cellular retinol-binding protein 1 (CRBP1). Specifically, an impaired RA signal through RARalpha in human breast epithelial cells triggers a repressive epigenetic domino effect, involving first RARbeta2 and second CRBP1. The phenotype acquired by breast epithelial cells clearly implies that the resistance to RA-mediated growth inhibition precedes the acquisition of morphological epithelial transformation, thus supporting the occurrence of sequential transcriptional silencing of first RARbeta2 and second CRBP1. The identification of this epigenetic network mechanistically linking RARbeta2 and CRBP1 transcription provides the basis for devising more accurate epigenetic tests for the prediction of breast cancer risk. PMID- 17079451 TI - Ephrin-A1 facilitates mammary tumor metastasis through an angiogenesis-dependent mechanism mediated by EphA receptor and vascular endothelial growth factor in mice. AB - Ephrin-A1, the prototypic ligand for EphA receptor tyrosine kinases, is overexpressed in vascularized tumors relative to normal tissue. Moreover, ephrin A1-Fc fusion proteins induce endothelial cell sprouting, migration, and assembly in vitro, and s.c. vascular remodeling in vivo. Based on these data, we hypothesized that native, membrane-bound ephrin-A1 regulates tumor angiogenesis and progression. We tested this hypothesis using a transplantable mouse mammary tumor model. Small interfering RNA-mediated ephrin-A1 knockdown in metastatic mammary tumor cells significantly diminishes lung metastasis without affecting tumor volume, invasion, intravasation, or lung colonization upon i.v. injection in vivo. Ephrin-A1 knockdown reduced tumor-induced endothelial cell migration in vitro and microvascular density in vivo. Conversely, overexpression of ephrin-A1 in nonmetastatic mammary tumor cells elevated microvascular density and vascular recruitment. Overexpression of ephrin-A1 elevated wild-type but not EphA2 deficient endothelial cell migration toward tumor cells, suggesting that activation of EphA2 on endothelial cells is one mechanism by which ephrin-A1 regulates angiogenesis. Furthermore, ephrin-A1 knockdown diminished, whereas overexpression of ephrin-A1 elevated, vascular endothelial growth factor (VEGF) levels in tumor cell-conditioned medium, suggesting that ephrin-A1-mediated modulation of the VEGF pathway is another mechanism by which membrane-tethered ephrin-A1 regulates angiogenic responses from initially distant host endothelium. These data suggest that ephrin-A1 is a proangiogenic signal, regulating VEGF expression and facilitating angiogenesis-dependent metastatic spread. PMID- 17079452 TI - p16INK4a modulates p53 in primary human mammary epithelial cells. AB - p16(INK4a) (p16) and p53 are tumor suppressor genes that are inactivated during carcinogenesis in many tumors. Here we show that p16 gene activity inversely modulates p53 status and function in primary human mammary epithelial cells. Reduced levels of p16 protein stabilize p53 protein through inhibition of proteolytic degradation, and this increase in p53 protein levels enhances the cellular response to radiation, represses proliferation, and transcriptionally activates downstream targets. Stabilization of p53 is mediated through the retinoblastoma/E2F/p14(ARF)/murine double minute-2 pathway. However, we have observed that p16 does not modulate p53 in fibroblasts, indicating a possible cell type-specific regulation of this pathway. PMID- 17079453 TI - Chromosomal breakpoints affecting immunoglobulin loci are recurrent in Hodgkin and Reed-Sternberg cells of classical Hodgkin lymphoma. AB - Chromosomal breakpoints affecting immunoglobulin (IG) loci are recurrent in many subtypes of B-cell lymphomas. However, despite the predominant B-cell origin of the Hodgkin and Reed-Sternberg (HRS) cells in classical Hodgkin lymphoma (cHL), the presence of chromosomal translocations in IG loci has not yet been systematically explored. Therefore, we have investigated a series of cHL for chromosomal breakpoints in the IGH (n = 230), IGL (n = 139), and IGK (n = 138) loci by interphase cytogenetics. Breakpoints in the IGH, IGL, or IGK locus were observed in the HRS cells of 26 of 149 (17%), 2 of 70, and 1 of 77 evaluable cHLs, respectively. The IG partners could be identified in eight cHLs and involved chromosomal bands 2p16 (REL), 3q27 (BCL6, two cases), 8q24.1 (MYC), 14q24.3, 16p13.1, 17q12, and 19q13.2 (BCL3/RELB). In 65 of 85 (76%) cHLs evaluable for an IGH triple-color probe, the HRS cells showed evidence for a (partial) deletion of the IGH constant region, suggesting the presence of class switch recombination (CSR). Furthermore, analyses with this probe in cases with IGH breakpoints indicated that at least part of them seem to be derived from CSR defects. Our results show that chromosomal breakpoints affecting the IG loci are recurrent in cHL. PMID- 17079454 TI - Activation of CDCA1-KNTC2, members of centromere protein complex, involved in pulmonary carcinogenesis. AB - We found cotransactivation of cell division associated 1 (CDCA1) and kinetochore associated 2 (KNTC2), members of the evolutionarily conserved centromere protein complex, in non-small cell lung carcinomas (NSCLC). Immunohistochemical analysis using lung cancer tissue microarray confirmed high levels of CDCA1 and KNTC2 proteins in the great majority of lung cancers of various histologic types. Their elevated expressions were associated with poorer prognosis of NSCLC patients. Knockdown of either CDCA1 or KNTC2 expression with small interfering RNA significantly suppressed growth of NSCLC cells. Furthermore, inhibition of their binding by a cell-permeable peptide carrying the CDCA1-derived 19-amino-acid peptide (11R-CDCA1(398-416)) that correspond to the binding domain to KNTC2 effectively suppressed growth of NSCLC cells. As our data imply that human CDCA1 and KNTC2 seem to fall in the category of cancer-testis antigens, and that their simultaneous up-regulation is a frequent and important feature of cell growth/survival of lung cancer, selective suppression of CDCA1 or KNTC2 activity and/or inhibition of the CDCA1-KNTC2 complex formation could be a promising therapeutic target for treatment of lung cancers. PMID- 17079455 TI - Low dietary folate initiates intestinal tumors in mice, with altered expression of G2-M checkpoint regulators polo-like kinase 1 and cell division cycle 25c. AB - Clinical reports have suggested that low dietary folate increases risk for colorectal cancer. Animal studies for investigation of folate and tumorigenesis have used carcinogen induction or mice with germ-line mutations. We have developed a new spontaneous tumor model in which mice, with or without a null allele in a key folate-metabolizing enzyme, methylenetetrahydrofolate reductase (Mthfr), develop intestinal tumors due to low dietary folate alone. On folate deficient diets, 12.5% of Mthfr(+/+) mice and 28.1% of Mthfr(+/-) mice developed tumors; mice on control diets were negative. Dietary and genotype effects on tumor development were significant. To investigate mechanisms of folate-dependent tumorigenesis, we examined levels of DNA damage and gene expression of two genes involved in DNA damage response and G(2)-M checkpoint regulation, polo-like kinase 1 (Plk1) and cell division cycle 25c (Cdc25c). Folate deficiency increased DNA damage and decreased expression of both genes (assessed by quantitative reverse transcription-PCR and immunofluorescence) in normal intestine compared with levels in mice on control diets. An immunofluorescence assay for CDC25c activity (phosphorylated CDC2) also found CDC25c activity to be decreased in folate-deficient normal intestine. In tumors, however, Plk1 and Cdc25c mRNA were found to be higher (11- and 3-fold, respectively) compared with normal intestine from folate-deficient mice; immunofluorescence studies of PLK1, CDC25c, and phosphorylated CDC2 supported these findings. Our data suggest that folate deficiency can initiate tumor development, that Mthfr mutation can enhance this phenomenon, and that altered expression of Plk1 and Cdc25c may contribute to folate-dependent intestinal tumorigenesis. PMID- 17079456 TI - Norepinephrine up-regulates the expression of vascular endothelial growth factor, matrix metalloproteinase (MMP)-2, and MMP-9 in nasopharyngeal carcinoma tumor cells. AB - Recent studies using ovarian cancer cells have shown that the catecholamine hormones norepinephrine (norepi) and epinephrine (epi) may influence cancer progression by modulating the expression of matrix metalloproteinases (MMP) and vascular endothelial growth factor (VEGF). The purpose of this study is to determine if the stress hormone norepi can influence the expression of MMP-2, MMP 9, and VEGF in nasopharyngeal carcinoma (NPC) tumors by using three NPC tumor cell lines. The NPC cell lines HONE-1, HNE-1, and CNE-1 were treated with norepi. The effects of norepi on MMP-2, MMP-9, and VEGF synthesis were measured by ELISA; functional MMP activity was measured by the invasive potential of the cells using a membrane invasion culture system whereas functional activity of VEGF was analyzed using a human umbilical vein endothelial cell tube formation assay. Norepi treatment increased MMP-2, MMP-9, and VEGF levels in culture supernatants of HONE-1 cells, which could be inhibited by the beta-blocker propranolol. Norepi induced the invasiveness of all NPC cell lines in a dose-dependent manner, which was blocked by CMT-3, an MMP inhibitor, and propranolol. Norepi stimulated the release of functional angiogenic VEGF by HONE-1 cells as well. Finally, HONE-1 cells were shown to express beta-adrenergic receptors as did seven of seven NPC biopsies examined. The data suggest that catecholamine hormones produced by the sympathetic-adrenal medullary axis may affect NPC tumor progression, in part, through modulation of key angiogenic cytokines. PMID- 17079458 TI - Mapping a sex hormone-sensitive gene determining female resistance to liver carcinogenesis in a congenic F344.BN-Hcs4 rat. AB - Hepatocellular carcinoma (HCC) is prevalent in human and rodent males. Hepatocarcinogenesis is controlled by various genes in susceptible F344 and resistant Brown Norway (BN) rats. B alleles at Hcs4 locus, on RNO16, control neoplastic nodule volume. We constructed the F344.BN-Hcs4 recombinant congenic strain (RCS) by introgressing a 4.41-cM portion of Hcs4 from BN strain in an isogenic F344 background. Preneoplastic and neoplastic lesions were induced by the "resistant hepatocyte" protocol. Eight weeks after initiation, lesion volume and positivity for proliferating cell nuclear antigen (PCNA) were much higher in lesions of F344 than BN rats of both sexes. These variables were lower in females than in males. Lesion volume and PCNA values of male RCS were similar to those of F344 rats, but in females corresponded to those of BN females. Carcinomatous nodules and HCC developed at 32 and 60 weeks, respectively, in male F344 and congenics and, rarely, in F344 females. BN and congenic females developed only eosinophilic/clear cells nodules. Gonadectomy of congenic males, followed by beta estradiol administration, caused a decrease in Ar expression, an increase in Er alpha expression, and development of preneoplastic lesions comparable to those from BN females. Administration of testosterone to gonadectomized females led to Ar increase and development of preneoplastic lesions as in F344 males. This indicates a role of homozygous B alleles at Hcs4 in the determination of phenotypic patterns of female RCS and presence at Hcs4 locus of a high penetrance gene(s), activated by estrogens and inhibited/unaffected by testosterone, conferring resistance to females in which the B alleles provide higher resistance. PMID- 17079459 TI - Effect of c-neu/ ErbB2 expression levels on estrogen receptor alpha-dependent proliferation in mammary epithelial cells: implications for breast cancer biology. AB - Mammary development and tumorigenesis are profoundly influenced by signaling pathways under the control of c-erbB2/c-neu and estrogen receptor alpha (ERalpha). Signaling through ERalpha is essential for ductal growth during puberty. In mice overexpressing wild-type c-neu in mammary epithelial cells, Tg (c-neu), ductal growth is impaired. An impeded signaling through ERalpha is also observed in a subset of human mammary tumors that overexpress erbB2. However, ductal growth is also impaired in the absence of c-neu in mouse mammary epithelial cells. To resolve this apparent paradox, we examined the relationship between c-neu expression and estrogen/ERalpha-dependent cell proliferation in pubertal Tg (c-neu). We report that proliferation in both terminal end buds and ducts is associated with ERalpha-positive cells, including those that coexpress c neu, and is abolished in the absence of circulating estradiol. Tg (c-neu) contains hyperplastic mammary ducts with high proliferative index and coexpression of both ERalpha and c-neu in the dividing cells. These findings suggest that c-neu promotes ERalpha-dependent proliferation, and that this is responsible for the presence of hyperplastic ducts. Some of the hyperplastic ducts have acinar structures, indicative of morphologic differentiation. These ducts have low proliferative index and accompanied by a vast decrease in proliferation of ERalpha-positive cells, including those that express c-neu. As such, c-neu has dual but opposing effects on ERalpha-dependent proliferation in mammary epithelial cells. Therefore, depending on the physiologic setting, ductal morphogenesis will be compromised both in the absence and overexpression of c neu, thus explaining the paradox. PMID- 17079457 TI - A role for mitochondrial dysfunction in perpetuating radiation-induced genomic instability. AB - Radiation-induced genomic instability (RIGI) manifests as a heritable increased rate of genetic alterations in the progeny of irradiated cells generations after the initial insult. The progeny can show an increased frequency of chromosomal translocations, deletions, mutations, micronuclei, and decreased plating efficiency. What perpetuates RIGI is unclear; however, persistently increased levels of reactive oxygen species (ROS) are frequently associated with genomically unstable clones. Furthermore, addition of free radical scavengers (e.g., DMSO, glycerol, and cationic thiol cysteamine) reduces the incidence of instability after irradiation, implicating a ROS-mediated role in RIGI induction. Because mitochondria are a major natural cellular source of ROS, we tested the hypothesis that mitochondrial dysfunction has a role in maintaining the elevated ROS levels in our irradiated, genetically unstable GM10115 Chinese hamster ovary cells. Amplex Red fluorometry measurements indicate that the relative contribution of uncoupler-sensitive mitochondrial hydrogen peroxide production to total cellular hydrogen peroxide generation is greater in unstable cells. Measurements of mitochondrial DNA levels and cell cytometric fluorescent measurements of Mitotracker Green FM indicate that differences in mitochondrial ROS production are not due to varying mitochondrial levels. However, mitochondrial respiration measured in digitonin-permeabilized cells is impaired in unstable clones. In addition, manganese superoxide dismutase, a major mitochondrial antioxidant enzyme, exhibits increased immunoreactivity but decreased enzyme activity in unstable clones, which along with decreased respiration rates may explain the increased levels of cellular ROS. These studies show that mitochondria from unstable cells are abnormal and likely contribute to the persistent oxidative stress in the unstable clones. PMID- 17079460 TI - Loss of Nkx2.8 deregulates progenitor cells in the large airways and leads to dysplasia. AB - Nkx2.8, a homeodomain transcription factor, has been characterized in liver cancer and in the developing central nervous system. We now show that this factor is also expressed in the lung, where it localizes in adults to a discrete population of tracheobronchial basal cells. To target the mouse gene, the first exon was replaced by a LacZ marker gene joined to the intact 5'-untranslated region. Marker expression was observed throughout the lower respiratory tract, beginning on E11 in a few cells of the distal lung buds. The region of expression then spread upward. By neonatal day 1, expression was greatest in the large airways and the Nkx2.8-/- mice exhibited generalized tracheobronchial hyperplasia. Bromodeoxyuridine (BrdUrd) labeling studies showed that a higher rate of bronchial cell proliferation persisted at 6 to 8 months. In adults, Nkx2.8 marker expression decreased with progressive differentiation into ciliated and secretory cells. The cell localizations and patterns of coexpression with BrdUrd and differentiation markers suggest a progenitor relationship: the cells that most strongly express Nkx2.8 seem to function as tracheobronchial stem cells. Moreover, Nkx2.8 acts to limit the number of these progenitor cells because the marker-expressing population was greatly expanded in Nkx2.8-/- mice. Increased proliferation and an altered progenitor relationship caused progressive bronchial pathology, which manifested as widespread dysplasia in the large airways of 1-year-old Nkx2.8-/- mice. PMID- 17079461 TI - Membrane ganglioside enrichment lowers the threshold for vascular endothelial cell angiogenic signaling. AB - Malignant tumor progression depends on angiogenesis, requiring vascular endothelial cell migration, and proliferation, triggered by tumor-derived vascular endothelial cell growth factor (VEGF). We show that gangliosides, which are actively shed by tumor cells and bind to normal cells in the tumor microenvironment, have the potential to sensitize vascular endothelial cells to respond to subthreshold levels of VEGF: Ganglioside enrichment of human umbilical vein vascular endothelial cells (HUVEC) caused very low, normally barely stimulatory, VEGF concentrations to trigger robust VEGF receptor dimerization and autophosphorylation, as well as activation of downstream signaling pathways, and cell proliferation and migration. Thus, by dramatically lowering the threshold for growth factor activation of contiguous normal stromal cells, shed tumor gangliosides may promote tumor progression by causing these normal cells to become increasingly autonomous from growth factor requirements by a process that we term tumor-induced progression of the microenvironment. PMID- 17079462 TI - Tumorigenic conversion of primary human esophageal epithelial cells using oncogene combinations in the absence of exogenous Ras. AB - To investigate pathways of human esophageal squamous cell transformation, we generated esophageal tumor cells using human telomerase- and SV40-immortalized primary esophageal epithelial cells (EPC2) by overexpression of selected combinations of oncogenes. H-Ras, c-Myc, or Akt, but not epidermal growth factor receptor (EGFR), induced transformed colonies in soft agar. By contrast, bioluminescence imaging of genetically altered immortalized esophageal cells revealed that Akt, EGFR, or H-Ras, but not c-Myc, resulted in tumor formation in immunodeficient mice. H-Ras-driven tumors showed highly tumorigenic phenotypes with 2.6 +/- 0.6 days for doubling, whereas Akt and EGFR tumors doubled every 9.5 +/- 1.6 and 6.1 +/- 1.2 days, respectively. H-Ras-driven tumors expressed the hypoxia-inducible factor target Glut1, whereas Akt- or EGFR-driven tumors had evidence of angiogenesis and no detectable Glut1 expression. Proliferation rates among these tumors were similar, but there was reduced apoptosis in the more aggressive H-Ras-driven tumors that also developed aneuploidy and multiple centrosomes. c-Myc overexpression did not result in tumorigenic conversion but introduction of Bcl-XL into c-Myc-expressing cells generated tumors. Although cytokeratin expression was typical of squamous carcinoma, gene expression profiling was done to compare the four different types of engineered tumors with human esophageal squamous cell carcinomas and adenocarcinomas. Interestingly, c Myc plus Bcl-XL transformants mimicked squamous carcinomas, whereas H-Ras-, EGFR , and Akt-driven tumors were similar to adenocarcinomas in their molecular profiles. These genetically engineered models may provide new platforms for understanding human esophagus cancer and may assist in the evaluation of new therapies. PMID- 17079463 TI - Disulfiram, a clinically used anti-alcoholism drug and copper-binding agent, induces apoptotic cell death in breast cancer cultures and xenografts via inhibition of the proteasome activity. AB - Disulfiram (DSF), a member of the dithiocarbamate family capable of binding copper and an inhibitor of aldehyde dehydrogenase, is currently being used clinically for the treatment of alcoholism. Recent studies have suggested that DSF may have antitumor and chemosensitizing activities, although the detailed molecular mechanisms remain unclear. Copper has been shown to be essential for tumor angiogenesis processes. Consistently, high serum and tissue levels of copper have been found in many types of human cancers, including breast, prostate, and brain, supporting the idea that copper could be used as a potential tumor-specific target. Here we report that the DSF-copper complex potently inhibits the proteasomal activity in cultured breast cancer MDA-MB-231 and MCF10DCIS.com cells, but not normal, immortalized MCF-10A cells, before induction of apoptotic cancer cell death. Furthermore, MDA-MB-231 cells that contain copper at concentrations similar to those found in patients, when treated with just DSF, undergo proteasome inhibition and apoptosis. In addition, when administered to mice bearing MDA-MB-231 tumor xenografts, DSF significantly inhibited the tumor growth (by 74%), associated with in vivo proteasome inhibition (as measured by decreased levels of tumor tissue proteasome activity and accumulation of ubiquitinated proteins and natural proteasome substrates p27 and Bax) and apoptosis induction (as shown by caspase activation and apoptotic nuclei formation). Our study shows that inhibition of the proteasomal activity can be achieved by targeting tumor cellular copper with the nontoxic compound DSF, resulting in selective apoptosis induction within tumor cells. PMID- 17079464 TI - A conserved RAS/mitogen-activated protein kinase pathway regulates DNA damage induced cell death postirradiation in Radelegans. AB - Although the epidermal growth factor receptor (EGFR) signaling pathway is overactive in more than half of human cancers and mediates resistance to cytotoxic therapy, the molecular mechanisms of EGFR pathway-mediated resistance have remained elusive in cancer research. This difficulty partly stems from the lack of tissue models enabling clear separation of the many forms of cell death that the downstream signaling pathways of EGFR affect. We have created a model in Caenorhabditis elegans of radiation-induced reproductive cell death ("Radelegans") in isolation of all other forms of cell death. We have employed Radelegans to genetically define the role of the EGFR signaling pathway in protection from reproductive cell death, the primary form of tumor stem or clonogen cell death postirradiation. We have found that the RAS/mitogen-activated protein kinase (MAPK) downstream signal transduction pathway of EGFR is critical for protection from reproductive cell death in Radelegans. In addition, we have shown that RAS/MAPK pathway signaling is genetically linear with the DNA damage response pathway and acts downstream of the DNA damage checkpoint in the radioresponse, implicating this pathway in DNA repair post-cytotoxic therapy. These findings support the hypothesis that enhanced repair is a mechanism of RAS/MAPK pathway-mediated resistance to cytotoxic therapy through its interaction with the DNA damage response pathway postirradiation. We postulate that these findings also help explain why current treatment strategies, based on the presumption that tumors have ineffective repair compared with normal tissues, are ineffective in EGFR/RAS/MAPK pathway-mediated tumors. Radelegans is a platform to further define the genetic basis of the radiation response in tissues. PMID- 17079465 TI - Expression of Wnt-5a is correlated with aggressiveness of gastric cancer by stimulating cell migration and invasion. AB - Wnt-5a is a representative ligand that activates a beta-catenin-independent pathway in the Wnt signaling. Although abnormal activation of beta-catenin dependent pathway is often observed in human cancer, the relationship between beta-catenin-independent pathway and tumorigenesis is not clear. We sought to clarify how Wnt-5a is involved in aggressiveness of gastric cancer. Abnormal expression of Wnt-5a was observed in 71 of 237 gastric cancer cases by means of immunohistochemistry. The positivity of Wnt-5a expression was correlated with advanced stages and poor prognosis of gastric cancer. Wnt-5a had the abilities to stimulate cell migration and invasion in gastric cancer cells. Wnt-5a activated focal adhesion kinase and small GTP-binding protein Rac, both of which are known to play a role in cell migration. Cell migration, membrane ruffling, and turnover of paxillin were suppressed in Wnt-5a knockdown cells. Furthermore, anti-Wnt-5a antibody suppressed gastric cancer cell migration. These results suggest that Wnt 5a stimulates cell migration by regulating focal adhesion complexes and that Wnt 5a is not only a prognostic factor but also a good therapeutic target for gastric cancer. PMID- 17079466 TI - Inappropriate activation of the androgen receptor by nonsteroids: involvement of the Src kinase pathway and its therapeutic implications. AB - The inappropriate activation of androgen receptor (AR) by nonsteroids is considered a potential mechanism in the emergence of hormone-refractory prostate tumors, but little is known about the properties of these "pseudoactivated" AR. Here, we present the first comprehensive analysis closely examining the properties of AR activated by the neuropeptide bombesin that distinguish it from androgen-activated AR. We show that bombesin-activated AR (a) is required for bombesin-induced growth of LNCaP cells, (b) has a transcriptional profile overlapping with, but not identical to, androgen-activated AR, (c) activates prostate-specific antigen by preferentially binding to its proximal promoter, and (d) assembles a distinct coactivator complex. Significantly, we found that Src kinase is critical for bombesin-induced AR-mediated activity and is required for translocation and transactivation of AR. Additionally, we identify c-Myc, a Src target gene, to be activated by bombesin and a potential coactivator of AR mediated activity specific to bombesin-induced signaling. Because Src kinase is often activated by other nonsteroids, such as other neuropeptides, growth factors, chemokines, and cytokines, our findings have general applicability and provide rationale for investigating the efficacy of the Src kinase pathway as a target for the prevention of relapsed prostate cancers. PMID- 17079467 TI - Membrane type-1 matrix metalloproteinase confers aneuploidy and tumorigenicity on mammary epithelial cells. AB - An elevated expression of membrane type-1 matrix metalloproteinase (MT1-MMP) is closely associated with multiple malignancies. Recently, we discovered that recycled MT1-MMP was trafficked along the tubulin cytoskeleton into the centrosomal compartment and cleaved the integral centrosomal protein pericentrin 2. These events correlated with the induction of chromosome instability and aneuploidy in nonmalignant Madine-Darby canine kidney cells. Accordingly, we hypothesized that MT1-MMP is an oncogene that promotes malignant transformation of normal cells rather than just an enzyme that supports growth of preexisting tumors. To prove our hypothesis, we transfected normal 184B5 human mammary epithelial cells with MT1-MMP (184B5-MT1 cells). MT1-MMP was colocalized with pericentrin in the centrosomal compartment and especially in the midbody of dividing cells. 184B5-MT1 cells acquired the ability to activate MMP-2, to cleave pericentrin, and to invade the Matrigel matrix. 184B5-MT1 cells exhibited aneuploidy, and they were efficient in generating tumors in the orthotopic xenograft model in immunodeficient mice. Because of the absence of tumor angiogenesis and the resulting insufficient blood supply, the tumors then regressed with significant accompanying necrosis. Gene array studies confirmed a significant up-regulation of oncogenes and tumorigenic genes but not the angiogenesis-promoting genes in 184B5-MT1 cells. We believe that our data point to a novel function of MT1-MMP in the initial stages of malignant transformation and to new and hitherto unknown transition mechanism from normalcy to malignancy. PMID- 17079468 TI - Identification and functional characterization of a novel unspliced transcript variant of HIC-1 in human cancer cells exposed to adverse growth conditions. AB - The wild-type p53 gene has been widely implicated in the regulation of hypermethylated in cancer-1 (HIC-1) transcription, a master growth regulatory gene with multiple promoters and, consequently, multiple alternatively spliced transcripts. We investigated the role of p53 (wild type and mutant, both endogenous and exogenous) in modulating the various HIC-1 transcripts. We discovered a novel unspliced HIC-1 transcript, identified as "f" in leukocytes and in the human cell lines U87MG (wild-type p53), U373MG (mutant p53), MCF-7 (wild-type p53), HeLa (p53 degraded by HPV18-E6 oncoprotein), and Saos-2 (p53 null). This transcript is initiated from a new transcription start site and has an intervening stop codon that would result in a possibly truncated 22-amino-acid polypeptide. When U87MG (wild-type p53) and MCF-7 cells (wild-type p53) were exposed to adverse growth conditions of serum starvation or treated with the chemotherapeutic agent cisplatin, cells underwent apoptosis and cell cycle arrest accompanied by increase in p53 and HIC-1 transcript levels. Although the increase of the HIC-1-spliced transcripts followed the increase of p53, increase in f transcript coincided with declining p53 and HIC-1 transcript and protein levels. Moreover, the levels of HIC-1 f transcript were not induced by exogenously transfected wild-type p53 in p53-mutated U373MG and p53-null Saos-2 cells, unlike the spliced transcripts that code for full-length HIC-1 protein. These findings suggest a working model wherein the status of f transcript, which is not under direct transcriptional control of wild-type p53, may influence the level of HIC-1 protein in cancer cells. PMID- 17079469 TI - A novel anticancer gold(III) dithiocarbamate compound inhibits the activity of a purified 20S proteasome and 26S proteasome in human breast cancer cell cultures and xenografts. AB - Although cisplatin has been used for decades to treat human cancer, some toxic side effects and resistance are observed. It has been suggested that gold(III) complexes, containing metal centers isoelectronic and isostructural to cisplatin, are promising anticancer drugs. Gold(III) dithiocarbamate complexes were shown to exhibit in vitro cytotoxicity, comparable with and even greater than cisplatin; however, the involved mechanism of action remained unknown. Because we previously reported that copper(II) dithiocarbamates are potent proteasome inhibitors, we hypothesized that gold(III) dithiocarbamate complexes could suppress tumor growth via direct inhibition of the proteasome activity. Here, for the first time, we report that a synthetic gold(III) dithiocarbamate (compound 2) potently inhibits the activity of a purified rabbit 20S proteasome and 26S proteasome in intact highly metastatic MDA-MB-231 breast cancer cells, resulting in the accumulation of ubiquitinated proteins and the proteasome target protein p27 and induction of apoptosis. The compound 2-mediated proteasome inhibition and apoptosis induction were completely blocked by addition of a reducing agent DTT or N-acetyl-L cysteine, showing that process of oxidation is required for proteasome inhibition by compound 2. Treatment of MDA-MB-231 breast tumor-bearing nude mice with compound 2 resulted in significant inhibition of tumor growth, associated with proteasome inhibition and massive apoptosis induction in vivo. Our findings reveal the proteasome as a primary target for gold(III) dithiocarbamates and support the idea for their potential use as anticancer therapeutics. PMID- 17079470 TI - Activating transcription factor 2 mediates matrix metalloproteinase-2 transcriptional activation induced by p38 in breast epithelial cells. AB - Mounting evidence suggests a role for matrix metalloproteinase (MMP)-2 in the malignant progression of breast cancer cells. We showed previously that H-Ras, but not N-Ras, induced invasion of MCF10A human breast epithelial cells through Rac-MKK3/6-p38 pathway resulted in MMP-2 up-regulation. Activation of p38 pathway by MKK6 caused a selective up-regulation of MMP-2. In this study, we aimed to elucidate the transcriptional regulation of MMP-2 by p38 pathway leading to the invasive phenotype of MCF10A cells. By using 5' deletion mutant constructs of MMP 2 promoter, we showed that deletion of the region containing activator protein-1 (AP-1) site caused the greatest reduction of MMP-2 promoter activity both in MKK6 and H-Ras-activated MCF10A cells, suggesting that the AP-1 binding site is critical for the MMP-2 promoter activation. DNA binding and transcriptional activities of AP-1 were increased by MKK6 or H-Ras as evidenced by electrophoretic mobility shift assay and luciferase assay using an AP-1-driven plasmid. By doing immunoinhibition assay and chromatin immunoprecipitation assay, we revealed the activating transcription factor (ATF) 2 as a transcription factor for MMP-2 gene expression through binding to the functional AP-1 site. Activation of ATF2, which depended on p38 activity, was crucial for MMP-2 promoter activity as well as induction of invasive and migrative phenotypes in MCF10A cells. This is the first report revealing ATF2 as an essential transcription factor linking MKK3/6-p38 signaling pathway to MMP-2 up-regulation, providing evidence for a direct role of ATF2 activation in malignant phenotypic changes of human breast epithelial cells. PMID- 17079471 TI - Combined stimulation with interleukin-18 and CpG induces murine natural killer dendritic cells to produce IFN-gamma and inhibit tumor growth. AB - Natural killer dendritic cells (NKDC) are a novel subtype of dendritic cells with natural killer (NK) cell properties. IFN-gamma is a pleiotropic cytokine that plays an important role in the innate immune response to tumors. Based on our previous finding that the combination of Toll-like receptor 9 ligand CpG and interleukin (IL)-4 stimulates NKDC to produce IFN-gamma, we hypothesized that NKDC are the major IFN-gamma-producing dendritic cell subtype and may play a significant role in the host antitumor response. We found that under several conditions in vitro and in vivo NKDC accounted for the majority of IFN-gamma production by murine spleen CD11c(+) cells. IL-18 alone induced NKDC to secrete IFN-gamma, and the combination of IL-18 and CpG resulted in a synergistic increase in IFN-gamma production, both in vitro and in vivo. NK cells made 26 fold less IFN-gamma under the same conditions in vitro, whereas dendritic cells produced a negligible amount. The mechanism of IFN-gamma secretion by NKDC depended on IL-12. NKDC selectively proliferated in vitro and in vivo in response to the combination of IL-18 and CpG. Systemic treatment with IL-18 and CpG reduced the number of B16F10 melanoma lung metastases. The mechanism depended on NK1.1(+) cells, as their depletion abrogated the effect. IL-18 and CpG activated NKDC provided greater tumor protection than NK cells in IFN-gamma(-/-) mice. Thus, NKDC are the major dendritic cell subtype to produce IFN-gamma. The combined use of IL-18 and CpG is a viable strategy to potentiate the antitumor function of NKDC. PMID- 17079472 TI - Ras induces chromosome instability and abrogation of the DNA damage response. AB - Ras mutations are frequent in thyroid tumors, the most common endocrine malignancy. The ability of Ras to transform thyroid cells is thought to rely on its mitogenic activity. Unexpectedly, acute expression of activated Ras in normal rat thyroid cells induced a DNA damage response, followed by apoptosis. Notably, a subpopulation of cells evaded apoptosis and emerged with features of transformation, including the loss of epithelial morphology, dedifferentiation, and the acquisition of hormone- and anchorage-independent proliferation. Strikingly, the surviving cells showed marked chromosomal instability. Acutely, Ras stimulated replication stress as evidenced by the induction of ataxia telangiectasia mutated and Rad3-related protein kinase (ATR) activity (Chk1 phosphorylation) and of gammaH2A.X, a marker of DNA damage. Despite the activation of a checkpoint, cells continued through mitosis in the face of DNA damage, resulting in an increase in cells harboring micronuclei, an indication of defects in chromosome segregation and other forms of chromosome damage. Cells that survived exposure to Ras continued to exhibit replication stress (ATR activation) but no longer exhibited gammaH2A.X or full activation of p53. When rechallenged with Ras or DNA-damaging agents, the surviving cells were more resistant to apoptosis than parental cells. These data show that acute expression of activated Ras is sufficient to induce chromosomal instability in the absence of other signals, and suggest that Ras-induced chromosomal instability arises as a consequence of defects in the processing of DNA damage. Hence, abrogation of the DNA damage response may constitute a novel mechanism for Ras transformation. PMID- 17079473 TI - Transgenic mouse model for rapid pharmacodynamic evaluation of antiandrogens. AB - Persistent androgen receptor signaling has been implicated as a critical factor in prostate cancer progression even at the hormone-refractory stage and provides strong rationale for developing novel androgen receptor antagonists. Traditional models for in vivo evaluation of antiandrogens are cumbersome because they rely on physiologic end points, such as the size of androgen-dependent tissues. Here, we describe a transgenic mouse (ARR2 Pb-Lux) that expresses luciferase specifically in the prostate in an androgen-dependent fashion. This signal is reduced by castration or by treatment with bicalutamide and can be quantified through noninvasive bioluminescent imaging. ARR2 Pb-Lux mice provide a novel method for rapid pharmacodynamic evaluation of novel pharmacologic compounds designed to inhibit androgen receptor signaling. PMID- 17079474 TI - Interleukin-6 contributes to growth in cholangiocarcinoma cells by aberrant promoter methylation and gene expression. AB - The association between chronic inflammation and the development and progression of malignancy is exemplified in the biliary tract where persistent inflammation strongly predisposes to cholangiocarcinoma. The inflammatory cytokine interleukin 6 (IL-6) enhances tumor growth in cholangiocarcinoma by altered gene expression via autocrine mechanisms. IL-6 can regulate the activity of DNA methyltransferases, and moreover, aberrant DNA methylation can contribute to carcinogenesis. We therefore investigated the effect of chronic exposure to IL-6 on methylation-dependent gene expression and transformed cell growth in human cholangiocarcinoma. The relationship between autocrine IL-6 pathways, DNA methylation, and transformed cell growth was assessed using malignant cholangiocytes stably transfected to overexpress IL-6. Treatment with the DNA methylation inhibitor 5-aza-2'-deoxycytidine decreased cell proliferation, growth in soft agar, and methylcytosine content of malignant cholangiocytes. However, this effect was not observed in IL-6-overexpressing cells. IL-6 overexpression resulted in the altered expression and promoter methylation of several genes, including the epidermal growth factor receptor (EGFR). EGFR promoter methylation was decreased and gene and protein expression was increased by IL-6. Thus, epigenetic regulation of gene expression by IL-6 can contribute to tumor progression by altering promoter methylation and gene expression of growth regulatory pathways, such as those involving EGFR. Moreover, enhanced IL-6 expression may decrease the sensitivity of tumor cells to therapeutic treatments using methylation inhibitors. These observations have important implications for cancer treatment and provide a mechanism by which persistent cytokine stimulation can promote tumor growth. PMID- 17079475 TI - Increased expression of tissue transglutaminase in pancreatic ductal adenocarcinoma and its implications in drug resistance and metastasis. AB - Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive neoplastic diseases and is virtually incurable. The molecular mechanisms that contribute to the intrinsic resistance of PDAC to various anticancer therapies are not well understood. Recently, we have observed that several drug-resistant and metastatic tumors and tumor cell lines expressed elevated levels of tissue transglutaminase (TG2). Because PDAC exhibits inherent resistance to various drugs, we determined the constitutive expression of TG2 in 75 PDAC and 12 PDAC cell lines. Our results showed that 42 of 75 (56%) PDAC tumor samples expressed higher basal levels of TG2 compared with the normal pancreatic ducts [odds ratio (OR), 2.439; P = 0.012]. The increased expression of TG2 in PDAC was strongly associated with nodal metastasis (OR, 3.400; P = 0.017) and lymphovascular invasion (OR, 3.055; P = 0.045). Increased expression of TG2 was also evident in all 12 cell lines examined. The elevated expression of TG2 in PDAC cell lines was associated with gemcitabine resistance and increased invasive potential. Overexpression of catalytically active or inactive (C(277)S mutant) TG2 induced focal adhesion kinase (FAK) activation and augmented invasive functions in the BxPC-3 cell line. Conversely, down-regulation of TG2 by small interfering RNA attenuated FAK phosphorylation. Immunoprecipitation and confocal microscopy data revealed that TG2 was associated with FAK protein in PDAC cells. The activated FAK colocalized with TG2 at focal adhesion points. These results show for the first time that elevated expression of TG2 can induce constitutive activation of FAK and thus may contribute to the development of drug resistance and invasive phenotypes in PDAC. PMID- 17079476 TI - Targeting cancer cells by novel engineered modular transporters. AB - A major problem in the treatment of cancer is the specific targeting of drugs to these abnormal cells. Ideally, such a drug should act over short distances to minimize damage to healthy cells and target subcellular compartments that have the highest sensitivity to the drug. We describe the novel approach of using modular recombinant transporters to target photosensitizers to the nucleus, where their action is most pronounced, of cancer cells overexpressing ErbB1 receptors. We have produced a new generation of the transporters consisting of (a) epidermal growth factor as the internalizable ligand module to ErbB1 receptors, (b) the optimized nuclear localization sequence of SV40 large T-antigen, (c) a translocation domain of diphtheria toxin as an endosomolytic module, and (d) the Escherichia coli hemoglobin-like protein HMP as a carrier module. The modules retained their functions within the transporter chimera: they showed high affinity interactions with ErbB1 receptors and alpha/beta-importin dimers and formed holes in lipid bilayers at endosomal pH. A photosensitizer conjugated with the transporter produced singlet oxygen and (*)OH radicals similar to the free photosensitizer. Photosensitizers-transporter conjugates have >3,000 times greater efficacy than free photosensitizers for target cells and were not photocytotoxic at these concentrations for cells expressing a few ErbB1 receptors per cell, in contrast to free photosensitizers. The different modules of the transporters, which are highly expressed and easily purified to retain full activity of each of the modules, are interchangeable, meaning that they can be tailored for particular applications. PMID- 17079477 TI - The urinary metabolite profile of the dietary carcinogen 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine is predictive of colon DNA adducts after a low-dose exposure in humans. AB - Epidemiologic evidence indicates that exposure to heterocyclic amines in the diet is an important risk factor for the development of colon cancer. Well-done cooked meats contain significant levels of heterocyclic amines, which have been shown to cause cancer in laboratory animals. To better understand the mechanisms of heterocyclic amine bioactivation in humans, the most mass abundant heterocyclic amine, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), was used to assess the relationship between PhIP metabolism and DNA adduct formation. Ten human volunteers where administered a dietary relevant dose of [(14)C]PhIP 48 to 72 hours before surgery to remove colon tumors. Urine was collected for 24 hours after dosing for metabolite analysis, and DNA was extracted from colon tissue and analyzed by accelerator mass spectrometry for DNA adducts. All 10 subjects were phenotyped for cytochrome P4501A2 (CYP1A2), N-acetyltransferase 2, and sulfotransferase 1A1 enzyme activity. Twelve PhIP metabolites were detected in the urine samples. The most abundant metabolite in all volunteers was N-hydroxy PhIP-N(2)-glucuronide. Metabolite levels varied significantly between the volunteers. Interindividual differences in colon DNA adducts levels were observed between each individual. The data showed that individuals with a rapid CYP1A2 phenotype and high levels of urinary N-hydroxy-PhIP-N(2)-glucuronide had the lowest level of colon PhIP-DNA adducts. This suggests that glucuronidation plays a significant role in detoxifying N-hydroxy-PhIP. The levels of urinary N-hydroxy PhIP-N(2)-glucuronide were negatively correlated to colon DNA adduct levels. Although it is difficult to make definite conclusions from a small data set, the results from this pilot study have encouraged further investigations using a much larger study group. PMID- 17079478 TI - Receptor-mediated DNA-targeted photoimmunotherapy. AB - We show the efficacy of a therapeutic strategy that combines the potency of a DNA binding photosensitizer, UV(A)Sens, with the tumor-targeting potential of receptor-mediated endocytosis. The photosensitizer is an iodinated bibenzimidazole, which, when bound in the minor groove of DNA and excited by UV(A) irradiation, induces cytotoxic lesions attributed to a radical species resulting from photodehalogenation. Although reminiscent of photochemotherapy using psoralens and UV(A) irradiation, an established treatment modality in dermatology particularly for the treatment of psoriasis and cutaneous T-cell lymphoma, a critical difference is the extreme photopotency of the iodinated bibenzimidazole, approximately 1,000-fold that of psoralens. This feature prompted consideration of combination with the specificity of receptor-mediated targeting. Using two in vitro model systems, we show the UV(A) cytotoxicity of iodo ligand/protein conjugates, implying binding of the conjugate to cell receptors, internalization, and degradation of the conjugate-receptor complex, with release and translocation of the ligand to nuclear DNA. For ligand transferrin conjugates, phototoxicity was inhibited by coincubation with excess native transferrin. Receptor-mediated UV(A)-induced cytotoxicity was also shown with the iodo ligand conjugate of an anti-human epidermal growth factor receptor monoclonal antibody, exemplifying the potential application of the strategy to other cancer-specific targets to thus improve the specificity of phototherapy of superficial lesions and for extracorporeal treatments. PMID- 17079479 TI - Potentiation of the effect of erlotinib by genistein in pancreatic cancer: the role of Akt and nuclear factor-kappaB. AB - The epidermal growth factor receptor (EGFR) is a target of new therapies in most nonhematologic cancers. EGFR blockade alone may not be sufficient for the control of growth and invasion of human pancreas cancer because of the independent activation of Akt and nuclear factor-kappaB (NF-kappaB). The expression of EGFR, Akt, and NF-kappaB was determined in six human pancreatic cancer cell lines. Selected cells for specific expression were treated with erlotinib, genistein, gemcitabine, or the combination. Growth inhibition was evaluated using 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and apoptosis was assayed by ELISA. EGFR, phosphorylated EGFR, phosphorylated Akt, and survivin expressions were determined by immunoblotting. Electrophoretic mobility shift assay was used to evaluate the DNA binding activity of NF-kappaB. Genistein significantly increased (P < 0.05) erlotinib-induced growth inhibition and apoptosis in BxPC-3, CAPAN-2, and AsPC-1 cells. In the BxPC-3 cell line, significant down-regulation of EGFR, phosphorylated Akt, NF-kappaB activation, and survivin was observed in the cells treated with the combination compared with the erlotinib-treated cells. In the HPAC and MIAPaCa cell line, no potentiation of the effects of erlotinib by genistein on cell growth or inhibition of the EGFR/Akt/NF-kappaB was observed. Genistein potentiated growth inhibition and apoptosis of the gemcitabine and erlotinib combination in COLO-357 cell line. Genistein potentiates the growth inhibition and apoptosis induced by erlotinib and gemcitabine in certain pancreatic cancer cells. Akt and NF-kappaB inhibition represents one of the mechanisms for the potentiation of erlotinib- and gemcitabine-induced cell death by genistein. PMID- 17079480 TI - Modulation of oncogenic transcription and alternative splicing by beta-catenin and an RNA aptamer in colon cancer cells. AB - Activated beta-catenin regulates the transcription of oncogenic target genes and is critical for tumorigenesis. Because nuclear functions are frequently coupled, we investigated whether it also has a role in alternative splicing of oncogenic genes. We showed that stabilized beta-catenin caused alternative splicing of estrogen receptor-beta pre-mRNA in colon cancer cells. To establish a direct role of beta-catenin in regulated splicing, we selected a high-affinity RNA aptamer that associated with beta-catenin in vivo. Nuclear localized aptamer inhibited beta-catenin-dependent transcription of cyclin D1 and c-myc in colon cancer cells; thus, cells stably expressing the aptamer exhibited cell cycle arrest and reduced tumor forming potential. Most significantly, the aptamer prevented the alternative splicing induced by stabilized beta-catenin. Taken together, our results establish that beta-catenin has an important role in both transcription and splicing, and that its action can be modulated by a high-affinity RNA aptamer. The RNA aptamer could be further developed as a specific inhibitor for cancer therapeutics. PMID- 17079481 TI - Characterization of GD2 peptide mimotope DNA vaccines effective against spontaneous neuroblastoma metastases. AB - Disialoganglioside GD2 is an established target for immunotherapy in neuroblastoma. We tested the hypothesis that active immunization against the glycolipid GD2 using DNA vaccines encoding for cyclic GD2-mimicking decapeptides (i.e., GD2 mimotopes) is effective against neuroblastoma. For this purpose, two GD2 peptide mimotopes (MA and MD) were selected based on docking experiments to anti-GD2 antibody ch14.18 (binding free energy: -41.23 kJ/mol for MA and -48.06 kJ/mol for MD) and Biacore analysis (K(d) = 12.3 x 10(-5) mol/L for MA and 5.3 x 10(-5) mol/L for MD), showing a higher affinity of MD over MA. These sequences were selected for DNA vaccine design based on pSecTag2-A (pSA) also including a T cell helper epitope. GD2 mimicry was shown following transfection of CHO-1 cells with pSA-MA and pSA-MD DNA vaccines, with twice-higher signal intensity for cells expressing MD over MA. Finally, these DNA vaccines were tested for induction of tumor protective immunity in a syngeneic neuroblastoma model following oral DNA vaccine delivery with attenuated Salmonella typhimurium (SL 7207). Only mice receiving the DNA vaccines revealed a reduction of spontaneous liver metastases. The highest anti-GD2 humoral immune response and natural killer cell activation was observed in mice immunized with the pSA-MD, a finding consistent with superior calculated binding free energy, dissociation constant, and GD2 mimicry potential for GD2 mimotope MD over MA. In summary, we show that DNA immunization with pSA-MD may provide a useful strategy for active immunization against neuroblastoma. PMID- 17079482 TI - Distinct roles for IFN regulatory factor (IRF)-3 and IRF-7 in the activation of antitumor properties of human macrophages. AB - When properly activated, macrophages can be tumoricidal, thus making them attractive additions to standard cancer therapies. To this end, tolerance and activity of human autologous IFN-gamma-activated macrophages, produced in large scale for clinical use (MAK cells), have been assessed in pilot trials in cancer patients. In the present study, we tested the hypothesis that activation of IFN regulatory factor (IRF)-3 and IRF-7, with subsequent type I IFN production, may be involved in the acquisition of new antitumor functions by macrophages. Adenoviral vectors were generated for the delivery of constitutively active forms of IRF-3 (Ad-IRF-3) or IRF-7 (Ad-IRF-7) into primary human macrophages. Cell death was observed in Ad-IRF-3-transduced macrophages, whereas Ad-IRF-7 transduced macrophages produced type I IFNs and displayed increased expression of genes encoding tumor necrosis factor (TNF)-related apoptosis-inducing ligand, interleukin (IL)-12, IL-15, and CD80, persisting for at least 96 hours. Expression of iNOS, TNF-alpha, FasL, IL-1, and IL-6 genes was unaltered by Ad-IRF 7 transduction. Interestingly, Ad-IRF-3 or Ad-IRF-7 transduction negatively regulated the transcription of protumorigenic genes encoding vascular endothelial growth factor and matrix metalloproteinase-2. Furthermore, Ad-IRF-7-transduced macrophages exerted a cytostatic activity on different cancer cell lines, including SK-BR-3, MCF-7, and COLO-205; the latter cells were shown previously to be insensitive to MAK cells. In conclusion, transduction of active forms of IRF-3 or IRF-7 differentially modulate the apoptotic and antitumor properties of primary macrophages, with active IRF-7 leading to the acquisition of novel antitumor effector functions. PMID- 17079483 TI - Eradication of tumor colonization and invasion by a B cell-specific immunotoxin in a murine model for human primary intraocular lymphoma. AB - Human primary intraocular lymphoma (PIOL) is predominantly a B cell-originated malignant disease with no appropriate animal models and effective therapies available. This study aimed to establish a mouse model to closely mimic human B cell PIOL and to test the therapeutic potential of a recently developed immunotoxin targeting human B-cell lymphomas. Human B-cell lymphoma cells were intravitreally injected into severe combined immunodeficient mice. The resemblance of this tumor model to human PIOL was examined by fundoscopy, histopathology, immunohistochemistry, and evaluated for molecular markers. The therapeutic effectiveness of immunotoxin HA22 was tested by injecting the drug intravitreally. Results showed that the murine model resembles human PIOL closely. Pathologic examination revealed that the tumor cells initially colonized on the retinal surface, followed by infiltrating through the retinal layers, expanding preferentially in the subretinal space, and eventually penetrating through the retinal pigment epithelium into the choroid. Several putative molecular markers for human PIOL were expressed in vivo in this model. Tumor metastasis into the central nervous system was also observed. A single intravitreal injection of immunotoxin HA22 after the establishment of the PIOL resulted in complete regression of the tumor. This is the first report of a murine model that closely mimics human B-cell PIOL. This model may be a valuable tool in understanding the molecular pathogenesis of human PIOL and for the evaluation of new therapeutic approaches. The results of B cell-specific immunotoxin therapy may have clinical implications in treating human PIOL. PMID- 17079484 TI - Androgens modulate expression of transcription intermediary factor 2, an androgen receptor coactivator whose expression level correlates with early biochemical recurrence in prostate cancer. AB - Prostate cancer is an androgen-dependent disease; metastatic prostate cancer is typically treated by androgen receptor (AR) blockade. Recurrence after androgen ablation and evidence that AR continues to play a role in many prostate cancers has led to an examination of other factors that potentiate AR activity. AR is a ligand-activated transcription factor whose activity is regulated not only by hormone but also by the levels of coactivators recruited by AR to facilitate transcription. We sought to assess the consequences of reducing expression of the transcription intermediary factor 2 (TIF2) coactivator on prostate cancer cell growth and AR action in cell lines to examine TIF2 expression in prostate cancer and to correlate expression with clinical outcome. Depletion of TIF2 reduced expression of AR-induced target genes and slowed proliferation of AR-dependent and AR-independent prostate cancer cells. Remarkably, we found that TIF2 expression is directly repressed by high levels of androgens in multiple AR expressing cell lines. Expression of a reporter containing 5'-flanking region of the TIF2 was repressed both by androgens and by the antagonist, Casodex. Expression of TIF2 correlates with biochemical (prostate-specific antigen) recurrence (P = 0.0136). In agreement with our in vitro findings, the highest expression of TIF2 was found in patients whose cancer relapsed after androgen ablation therapy, supporting the idea that AR blockade might activate pathways that lead to stimulation of AR-dependent and AR-independent proliferation of prostate epithelium. The elevated expression of TIF2 at low hormone levels likely aids in inducing AR activity under these conditions; treatment with Casodex has the potential to counteract this induction. PMID- 17079485 TI - PRKAR1A inactivation leads to increased proliferation and decreased apoptosis in human B lymphocytes. AB - The multiple neoplasia syndrome Carney complex (CNC) is caused by heterozygote mutations in the gene, which codes for the RIalpha regulatory subunit (PRKAR1A) of protein kinase A. Inactivation of PRKAR1A and the additional loss of the normal allele lead to tumors in CNC patients and increased cyclic AMP signaling in their cells, but the oncogenetic mechanisms in affected tissues remain unknown. Previous studies suggested that PRKAR1A down-regulation may lead to increased mitogen-activated protein kinase (MAPK) signaling. Here, we show that, in lymphocytes with PRKAR1A-inactivating mutations, there is increased extracellular signal-regulated kinase (ERK) 1/2 and B-raf phosphorylation and MAPK/ERK kinase 1/2 and c-Myc activation, whereas c-Raf-1 is inhibited. These changes are accompanied by increased cell cycle rates and decreased apoptosis that result in an overall net gain in proliferation and survival. In conclusion, inactivation of PRKAR1A leads to widespread changes in molecular pathways that control cell cycle and apoptosis. This is the first study to show that human cells with partially inactivated RIalpha levels have increased proliferation and survival, suggesting that loss of the normal allele in these cells is not necessary for these changes to occur. PMID- 17079486 TI - Short hairpin RNA knockdown of the androgen receptor attenuates ligand independent activation and delays tumor progression. AB - Progression to androgen independence is the lethal end stage of prostate cancer. We used expression of androgen receptor (AR)-targeted short hairpin RNAs (shRNA) to directly test the requirement for AR in ligand-independent activation of androgen-regulated genes and hormone-independent tumor progression. Transient transfection of LNCaP human prostate cancer cells showed that AR shRNA decreased R1881 induction of the prostate-specific antigen (PSA)-luciferase reporter by 96%, whereas activation by forskolin, interleukin-6, or epidermal growth factor was inhibited 48% to 75%. Whereas the antiandrogen bicalutamide provided no further suppression, treatment with the mitogen-activated protein kinase (MAPK) inhibitor U0126 completely abrogated the residual activity, indicating a MAPK dependent, AR-independent pathway for regulating the PSA promoter. Expression of doxycycline-inducible AR shRNA expression in LNCaP cells resulted in decreased levels of AR and PSA as well as reduced proliferation in vitro. When these cells were grown as xenografts in immunocompromised mice, induction of AR shRNA decreased serum PSA to below castration nadir levels and significantly retarded tumor growth over the entire 55-day experimental period. This is the first demonstration that, by inducibly suppressing AR expression in vivo, there is an extensive delay in progression to androgen independence as well as a dramatic inhibition of tumor growth and decrease in serum PSA, which exceeds that seen with castration alone. Based on these findings, we propose that suppressing AR expression may provide superior therapeutic benefit in reducing tumor growth rate than castration and may additionally be very effective in delaying progression to androgen independence. PMID- 17079487 TI - Examination of a CpG island methylator phenotype and implications of methylation profiles in solid tumors. AB - The CpG island methylator phenotype (CIMP), thoroughly described in colorectal cancer and to a lesser extent in other solid tumors, is important in understanding epigenetics in carcinogenesis and may be clinically useful for classification of neoplastic disease. Therefore, we investigated whether this putative phenotype exists in exposure-related solid tumors, where somatic gene alterations and enhanced clonal growth are selected for by carcinogens, and examined the ability of methylation profiles to classify malignant disease. We studied promoter hypermethylation of 16 tumor suppressor genes and 3 MINT loci (acknowledged classifiers of CIMP) in 344 bladder cancers, 346 head and neck squamous cell carcinomas (HNSCC), 146 non-small-cell lung cancer (NSCLC), and 71 malignant pleural mesotheliomas (MPM). We employed rigorous statistical methods to examine the distribution of promoter methylation and the usefulness of these profiles for disease classification. In bladder cancer, HNSCC, and NSCLC, there was a significant correlation (P < 0.0001) between methylation of the three MINT loci and methylation index, although the distribution of methylated loci varied significantly across these disease. Although there was a significant (P < 0.001) association between gene methylation profile and disease, rates of misclassification of each disease by their methylation profile ranged from 28% to 32%, depending on the classification scheme used. These data suggest that a form of CIMP exists in these solid tumors, although its etiology remains elusive. Whereas the gene profiles of hypermethylation among examined loci could not unequivocally distinguish disease type, the existence of CIMP and the relative preponderance of hypermethylation in these cancers suggest that methylation analysis may be clinically useful as a targeted screening tool. PMID- 17079488 TI - uPAR and HER-2 gene status in individual breast cancer cells from blood and tissues. AB - Overexpression of urokinase plasminogen activator system or HER-2 (erbB-2) in breast cancer is associated with a poor prognosis. HER-2 overexpression is caused by HER-2 gene amplification. The anti-HER-2 antibody trastuzumab significantly improves clinical outcome for HER2-positive breast cancer. Drugs that target the uPA system are in early clinical trials. The aims of this study were to determine whether urokinase plasminogen activator receptor (uPAR) gene amplification occurs and whether analysis of individual tumor cells (TCs) in the blood or tissue can add information to conventional pathological analysis that could help in diagnosis and treatment. Analysis of individual TCs indicates that uPAR amplification occurs in a significant portion of primary breast cancers and also circulating tumor cells (CTCs) from patients with advanced disease. There was complete concordance between touch preps (TPs) and conventional pathological examination of HER-2 and uPAR gene status in primary tumors. There was also excellent concordance of HER-2 gene status between primary tumors and CTCs provided that acquisition of HER-2 gene amplification in CTCs was taken into account. Unexpectedly, gene amplification of HER-2 and uPAR occurred most frequently in the same TC and patient, suggesting a biological bias and potential advantage for coamplification. Expression of HER-2 and uPAR in primary tumors predicted gene status in 100 and 92% of patients, respectively. PMID- 17079489 TI - Cognitive cladistics and cultural override in Hominid spatial cognition. AB - Current approaches to human cognition often take a strong nativist stance based on Western adult performance, backed up where possible by neonate and infant research and almost never by comparative research across the Hominidae. Recent research suggests considerable cross-cultural differences in cognitive strategies, including relational thinking, a domain where infant research is impossible because of lack of cognitive maturation. Here, we apply the same paradigm across children and adults of different cultures and across all nonhuman great ape genera. We find that both child and adult spatial cognition systematically varies with language and culture but that, nevertheless, there is a clear inherited bias for one spatial strategy in the great apes. It is reasonable to conclude, we argue, that language and culture mask the native tendencies in our species. This cladistic approach suggests that the correct perspective on human cognition is neither nativist uniformitarian nor "blank slate" but recognizes the powerful impact that language and culture can have on our shared primate cognitive biases. PMID- 17079490 TI - Podocin and MEC-2 bind cholesterol to regulate the activity of associated ion channels. AB - The prohibitin (PHB)-domain proteins are membrane proteins that regulate a variety of biological activities, including mechanosensation, osmotic homeostasis, and cell signaling, although the mechanism of this regulation is unknown. We have studied two members of this large protein family, MEC-2, which is needed for touch sensitivity in Caenorhabditis elegans, and Podocin, a protein involved in the function of the filtration barrier in the mammalian kidney, and find that both proteins bind cholesterol. This binding requires the PHB domain (including palmitoylation sites within it) and part of the N-terminally adjacent hydrophobic domain that attaches the proteins to the inner leaflet of the plasma membrane. By binding to MEC-2 and Podocin, cholesterol associates with ion channel complexes to which these proteins bind: DEG/ENaC channels for MEC-2 and TRPC channels for Podocin. Both the MEC-2-dependent activation of mechanosensation and the Podocin-dependent activation of TRPC channels require cholesterol. Thus, MEC-2, Podocin, and probably many other PHB-domain proteins by binding to themselves, cholesterol, and target proteins regulate the formation and function of large protein-cholesterol supercomplexes in the plasma membrane. PMID- 17079491 TI - Individuals homozygous for the age-related macular degeneration risk-conferring variant of complement factor H have elevated levels of CRP in the choroid. AB - Polymorphisms in the complement factor H gene (CFH) are associated with a significantly increased risk for, or protection against, the development of age related macular degeneration (AMD). The most documented risk-conferring single nucleotide polymorphism results in a tyrosine-to-histidine substitution at position 402 (Y402H) of the CFH protein. In this work, we examined the ocular distributions and relative abundance of CFH, several CFH-binding proteins, and abundant serum proteins in the retinal pigmented epithelium (RPE), Bruch's membrane, and choroid (RPE-choroid) in CFH homozygotes possessing either the "at risk" 402HH or "normal" 402YY variants. Although CFH immunoreactivity is high in the choroid and in drusen, no differences in CFH-labeling patterns between genotypes are apparent. In contrast, at-risk individuals have significantly higher levels of the CFH-binding protein, C-reactive protein (CRP), in the choroidal stroma. Immunoblots confirm that at-risk individuals have approximately 2.5-fold higher levels of CRP in the RPE-choroid; no significant differences in the levels of CFH or other serum proteins are detected. Similarly, we find no differences in CFH transcription levels in the RPE-choroid nor evidence for local ocular CRP transcription. Increased levels of CRP in the choroid may reflect a state of chronic inflammation that is a by-product of attenuated CFH complement inhibitory activity in those who possess the CFH at-risk allele. Because the CRP binding site in CFH lies within the domain containing the Y402H polymorphism, it is also possible that the AMD risk-conferring allele alters the binding properties of CFH, thereby leading to choroidal CRP deposition, contributing to AMD pathogenesis. PMID- 17079493 TI - Structure of DNA polymerase beta with a benzo[c]phenanthrene diol epoxide adducted template exhibits mutagenic features. AB - We have determined the crystal structure of the human base excision repair enzyme DNA polymerase beta (Pol beta) in complex with a 1-nt gapped DNA substrate containing a template N2-guanine adduct of the tumorigenic (-) benzo[c]phenanthrene 4R,3S-diol 2S,1R-epoxide in the gap. Nucleotide insertion opposite this adduct favors incorrect purine nucleotides over the correct dCMP and hence can be mutagenic. The structure reveals that the phenanthrene ring system is stacked with the base pair immediately 3' to the modified guanine, thereby occluding the normal binding site for the correct incoming nucleoside triphosphate. The modified guanine base is displaced downstream and prevents the polymerase from achieving the catalytically competent closed conformation. The incoming nucleotide binding pocket is distorted, and the adducted deoxyguanosine is in a syn conformation, exposing its Hoogsteen edge, which can hydrogen-bond with dATP or dGTP. In a reconstituted base excision repair system, repair of a deaminated cytosine (i.e., uracil) opposite the adducted guanine was dramatically decreased at the Pol beta insertion step, but not blocked. The efficiency of gap filling dCMP insertion opposite the adduct was diminished by >6 orders of magnitude compared with an unadducted templating guanine. In contrast, significant misinsertion of purine nucleotides (but not dTMP) opposite the adducted guanine was observed. Pol beta also misinserts a purine nucleotide opposite the adduct with ungapped DNA and exhibits limited bypass DNA synthesis. These results indicate that Pol beta-dependent base excision repair of uracil opposite, or replication through, this bulky DNA adduct can be mutagenic. PMID- 17079494 TI - A critical role for I kappaB kinase alpha in the development of human and mouse squamous cell carcinomas. AB - IKK (I kappaB kinase) alpha is essential for embryonic skin development in mice. Mice deficient in IKKalpha display markedly hyperplasic epidermis that lacks terminal differentiation, and they die because of this severely impaired skin. However, the function of IKKalpha in human skin diseases remains largely unknown. To shed light on the role of IKKalpha in human skin diseases, we examined IKKalpha expression and Ikkalpha mutations in human squamous cell carcinomas (SCCs). We found a marked reduction in IKKalpha expression in poorly differentiated human SCCs and identified Ikkalpha mutations in exon 15 of Ikkalpha in eight of nine human SCCs, implying that IKKalpha is involved in development of this human skin cancer. Furthermore, in a chemical carcinogen induced skin carcinogenesis setting, mice overexpressing human IKKalpha in the epidermis under the control of a truncated loricrin promoter developed significantly fewer SCCs and metastases than did wild-type mice. The IKKalpha transgene altered the skin microenvironment conditions, leading to elevated terminal differentiation in the epidermis, reduced mitogenic activity in the epidermis, and decreased angiogenic activity in the skin stroma. Thus, overexpression of IKKalpha in the epidermis antagonized chemical carcinogen induced mitogenic and angiogenic activities, repressing tumor progression and metastases. PMID- 17079495 TI - Genetic profiling of the oral microbiota associated with severe early-childhood caries. AB - The determination of the composition of the microbial community in the oral cavity is usually based on cultivation methods; however, nearly half of the bacteria in the saliva and the dental plaque are not cultivable. In this study, we evaluated the difference in oral microbial diversity between children with severe early-childhood caries (S-ECC) and caries-free (CF) controls by means of a cultivation-independent approach called denaturing gradient gel electrophoresis (DGGE). Pooled dental plaque samples were collected from 20 children aged 2 to 8 years. Total microbial genomic DNA was isolated from those subjects, and a portion of the 16S rRNA gene locus was PCR amplified by using universal primers. We observed that the mean species richness of the bacterial population was greater in the CF children (n = 12) (42 +/- 3.7) than in the S-ECC children (n = 8) (35 +/- 4.3); the difference was statistically significant (P = 0.005). The overall diversity of plaque samples as measured by the Shannon index was 3.5 for the S-ECC group and 3.7 for the CF group (P = 0.004). Differences in DGGE profiles were distinguished on the basis of a cluster analysis. Sequence analysis of excised DGGE bands consisted of 2.7 phylotypes, on average. After adjusting for the number of observed bands, we estimated that the S-ECC group exhibited 94.5 total phylotypes and that the CF group exhibited 113.4. These results suggest that the microbial diversity and complexity of the microbial biota in dental plaque are significantly less in S-ECC children than in CF children. PMID- 17079496 TI - Rapid molecular strategy for filovirus detection and characterization. AB - Filoviruses have the capacity to cause lethal outbreaks of hemorrhagic fever in primates. Here we present a simple consensus reverse transcription-PCR method for filovirus recognition and characterization and demonstrate its utility with all known filovirus strains. Phylogenetic assignment is achieved by automated web based sequence analysis of amplification products. PMID- 17079497 TI - Comparison between nasal swabs and nasopharyngeal aspirates for, and effect of time in transit on, isolation of Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, and Moraxella catarrhalis. AB - We assessed the impact of the use of nasal swabs or nasopharyngeal aspirates and the time from specimen collection to storage at -70 degrees C on bacterial isolation. Haemophilus influenzae was isolated significantly less often from swabs than from nasopharyngeal aspirates. Samples in transit for >3 days were half as likely to grow Streptococcus pneumoniae and H. influenzae as those in transit for < or =3 days. There was no statistically significant difference for either Moraxella catarrhalis or Staphylococcus aureus. PMID- 17079492 TI - Evaluating alternative hypotheses for the early evolution and diversification of ants. AB - Ants are the world's most diverse and ecologically dominant eusocial organisms. Resolving the phylogeny and timescale for major ant lineages is vital to understanding how they achieved this success. Morphological, molecular, and paleontological studies, however, have presented conflicting views on early ant evolution. To address these issues, we generated the largest ant molecular phylogenetic data set published to date, containing approximately 6 kb of DNA sequence from 162 species representing all 20 ant subfamilies and 10 aculeate outgroup families. When these data were analyzed with and without outgroups, which are all distantly related to ants and hence long-branched, we obtained conflicting ingroup topologies for some early ant lineages. This result casts strong doubt on the existence of a poneroid clade as currently defined. We compare alternate attachments of the outgroups to the ingroup tree by using likelihood tests, and find that several alternative rootings cannot be rejected by the data. These alternatives imply fundamentally different scenarios for the early evolution of ant morphology and behavior. Our data strongly support several notable relationships within the more derived formicoid ants, including placement of the enigmatic subfamily Aenictogitoninae as sister to Dorylus army ants. We use the molecular data to estimate divergence times, employing a strategy distinct from previous work by incorporating the extensive fossil record of other aculeate Hymenoptera as well as that of ants. Our age estimates for the most recent common ancestor of extant ants range from approximately 115 to 135 million years ago, indicating that a Jurassic origin is highly unlikely. PMID- 17079498 TI - Role of large sequence polymorphisms (LSPs) in generating genomic diversity among clinical isolates of Mycobacterium tuberculosis and the utility of LSPs in phylogenetic analysis. AB - Mycobacterium tuberculosis strains contain different genomic insertions or deletions called large sequence polymorphisms (LSPs). Distinguishing between LSPs that occur one time versus ones that occur repeatedly in a genomic region may provide insights into the biological roles of LSPs and identify useful phylogenetic markers. We analyzed 163 clinical M. tuberculosis isolates for 17 LSPs identified in a genomic comparison of M. tuberculosis strains H37Rv and CDC1551. LSPs were mapped onto a single-nucleotide polymorphism (SNP)-based phylogenetic tree created using nine novel SNP markers that were found to reproduce a 212-SNP-based phylogeny. Four LSPs (group A) mapped to a single SNP tree segment. Two LSPs (group B) and 11 LSPs (group C) were inferred to have arisen independently in the same genomic region either two or more than two times, respectively. None of the group A LSPs but one group B LSP and five group C LSPs were flanked by IS6110 sequences in the references strains. Genes encoding members of the proline-glutamic acid or proline-proline-glutamic acid protein families were present only in group B or C LSPs. SNP- versus LSP-based phylogenies were also compared. We classified each isolate into 58 LSP types by using a separate LSP-based phylogenetic analysis and mapped the LSP types onto the SNP tree. LSPs often assigned isolates to the correct phylogenetic lineage; however, significant mistakes occurred for 6/58 (10%) of the LSP types. In conclusion, most LSPs occur in genomic regions that are prone to repeated insertion/deletion events and were responsible for an unexpectedly high degree of genomic variation in clinical M. tuberculosis. Group B and C LSPs may represent polymorphisms that occur due to selective pressure and affect the phenotype of the organism, while group A LSPs are preferable phylogenetic markers. PMID- 17079499 TI - Probe hybridization array typing: a binary typing method for Escherichia coli. AB - The ability to distinguish between Escherichia coli strains is critical for outbreak investigations. Binary typing, based on the presence or absence of genetic material, provides a high-throughput alternative to gel- and PCR-based typing techniques that generate complex banding patterns and lack uniform interpretation criteria. We developed, validated, and determined the discriminatory power of an E. coli binary typing method, probe hybridization array typing (PHAT). In PHAT, the absence or presence of genetic material is identified by using DNA hybridization to produce a reproducible and portable fingerprint for each genome. PHAT probes were generated from genome subtractive hybridization experiments. We PHAT typed the ECOR collection of strains from a variety of geographical locations, and 33 rectal E. coli strains selected from college-aged women with urinary tract infection. In the set of 33 human rectal strains, the discriminatory power of PHAT (98%) equaled that of multilocus sequence typing (MLST) and pulsed-field gel electrophoresis. However, for ECOR strains, which include nonhuman strains, the current set of PHAT probes was less discriminating than MLST, ribotyping, and enterobacterial repetitive intergenic consensus sequence PCR (80% versus 97, 92, and 97%, respectively). When we limited the analysis to ECOR strains of B2 and D lineage, which are associated with human infection, current PHAT probes were highly discriminatory (94%). PHAT can be applied in a high-throughput format (i.e., "library on a slide"), the discriminatory ability can be varied based on the probe set, and PHAT is readily adapted to other bacterial species with high variation in genetic content. PMID- 17079500 TI - Helicobacter cinaedi cellulitis and bacteremia in immunocompetent hosts after orthopedic surgery. AB - At various times after orthopedic operations (more than a few weeks, with an average of 29.9 days), 11 patients had a sudden onset of high temperature (average 38.9 degrees C) and local cellulitis at different sites on the operated sides. The wounds had completely healed, without complicated infections, when the cellulitis occurred. The clinical picture of cellulitis in all patients was atypical: diffuse salmon-pink skin color, local heat, swelling, spontaneous pain, and tenderness but no eruptions. No patient had any underlying immunocompromising conditions or had been given immunosuppressive agents. Gram-negative spiral bacteria were isolated from blood cultures and were identified as Helicobacter cinaedi on the basis of 16S rRNA gene sequencing and DNA-DNA hybridization using standard strains. By means of phylogenetic analysis, we divided these clinical isolates into two clones. The H. cinaedi strain isolated via fecal cultures from two patients without intestinal symptoms was the same clone as the blood isolate. All isolates were quite susceptible to various antibiotics, and clinical and inflammatory symptoms of bacteremia and cellulitis improved after treatment with penicillins and cephalosporins. A relatively high incidence of recurrence of the same disease was observed, however. Almost all patients responded immunologically to the infection, as evidenced by the production of serum antibody against H. cinaedi. We thus suggest that H. cinaedi should not be regarded as simply an opportunistic pathogen but that it may be a pathogen in immunocompetent hosts and may cause infections together with bacteremia and cellulitis. PMID- 17079501 TI - Use of fluconazole as a surrogate marker to predict susceptibility and resistance to voriconazole among 13,338 clinical isolates of Candida spp. Tested by clinical and laboratory standards institute-recommended broth microdilution methods. AB - Clinical laboratories frequently face the problem of delayed availability of commercially prepared approved reagents for performing susceptibility testing of new antimicrobials. Although this problem is encountered more often with antibacterial agents, it is also an issue with antifungal agents. A current example is voriconazole, a new triazole antifungal with an expanded spectrum and potency against Candida spp., Aspergillus spp., and other opportunistic fungal pathogens. The present study addresses the use of fluconazole as a surrogate marker to predict the susceptibility of Candida spp. to voriconazole. Reference broth microdilution MIC results for 13,338 strains of Candida spp. isolated from more than 200 medical centers worldwide were used. Voriconazole MICs and interpretive categories (susceptible, < or =1 microg/ml; susceptible dose dependent, 2 microg/ml; resistant, > or =4 microg/ml) were compared with those of fluconazole by regression statistics and error rate bounding analyses. For all 13,338 isolates, the absolute categorical agreement was 91.6% (false susceptible or very major error [VME], 0.0%). Since voriconazole is 16- to 32-fold more potent than fluconazole, the performance of fluconazole as a surrogate marker for voriconazole susceptibility was improved by designating those isolates with fluconazole MICs of < or =32 microg/ml as being susceptible to voriconazole, resulting in a categorical agreement of 97% with 0.1% VME. Clinical laboratories performing antifungal susceptibility testing of fluconazole against Candida spp. can reliably use these results as surrogate markers until commercial FDA-approved voriconazole susceptibility tests become available. PMID- 17079503 TI - Visual diagnosis: a child who has acute onset of unusual skin lesions and edema. PMID- 17079502 TI - Effects of phenotype and genotype on methods for detection of extended-spectrum beta-lactamase-producing clinical isolates of Escherichia coli and Klebsiella pneumoniae in Norway. AB - Consecutive clinical isolates of Escherichia coli (n = 87) and Klebsiella pneumoniae (n = 25) with reduced susceptibilities to oxyimino-cephalosporins (MICs > 1 mg/liter) from 18 Norwegian laboratories during March through October 2003 were examined for bla(TEM/SHV/CTX-M) extended-spectrum-beta-lactamase (ESBL) genes, oxyimino-cephalosporin MIC profiles, ESBL phenotypes (determined by the ESBL Etest and the combined disk and double-disk synergy [DDS] methods), and susceptibility to non-beta-lactam antibiotics. Multidrug-resistant CTX-M-15-like (n = 23) and CTX-M-9-like (n = 15) ESBLs dominated among the 50 ESBL-positive E. coli isolates. SHV-5-like (n = 9) and SHV-2-like (n = 4) ESBLs were the most prevalent in 19 ESBL-positive K. pneumoniae isolates. Discrepant ESBL phenotype test results were observed for one major (CTX-M-9) and several minor (TEM-128 and SHV-2/-28) ESBL groups and in SHV-1/-11-hyperproducing isolates. Negative or borderline ESBL results were observed when low-MIC oxyimino-cephalosporin substrates were used to detect clavulanic acid (CLA) synergy. CLA synergy was detected by the ESBL Etest and the DDS method but not by the combined disk method in SHV-1/-11-hyperproducing strains. The DDS method revealed unexplained CLA synergy in combination with aztreonam and cefpirome in three E. coli strains. The relatively high proportion of ESBL-producing E. coli organisms with a low ceftazidime MIC in Norway emphasizes that cefpodoxime alone or both cefotaxime and ceftazidime should be used as substrates for ESBL detection. PMID- 17079504 TI - Commentary: the value of the ECG in the preparticipation sports physical examination: the Italian experience. PMID- 17079505 TI - Intelligence (IQ) testing. PMID- 17079506 TI - Breastfeeding: the essential principles. PMID- 17079507 TI - Consultation with the specialist: cardiovascular preparticipation sports screening. PMID- 17079508 TI - Index of suspicion. PMID- 17079509 TI - Corneal abrasions. PMID- 17079510 TI - Ampicillin and amoxicillin. PMID- 17079511 TI - Cocaine. PMID- 17079512 TI - The timing of neuronal development in adult hippocampal neurogenesis. AB - The granule cell layer (GCL) of the adult dentate gyrus (DG) is a heterogeneous structure formed by neurons of different ages because a significant proportion of neurons continues to be generated throughout life. The subgranular zone of the DG contains neural progenitor cells (NPCs) that divide, differentiate, and migrate to produce functional dentate granule cells (DGCs) that become incorporated into the existing hippocampal circuitry. New available tools to identify adult-born neurons in live and fixed brain sections have allowed the transition from NPC to functional neuron to be characterized in great detail. Maturation of the neuronal phenotype includes changes in membrane excitability and morphology as well as the establishment of appropriate connectivity within the existing circuits, a process that lasts several weeks. The events leading to neuronal maturation share many of the features of the developing brain, and electrical activity is emerging as a key modulator of neuronal development in the adult DG. The underlying mechanisms are now beginning to be understood. PMID- 17079513 TI - Microtubule: a common target for parkin and Parkinson's disease toxins. AB - Parkinson's disease (PD) is characterized by the selective loss of nigral dopaminergic (DA) neurons, which have long axons enriched with microtubules. Depolymerization of microtubules by PD toxins such as rotenone disrupts vesicular transport. The ensuing accumulation of vesicles in the cell body leads to increased cytosolic concentration of dopamine due to leakage of the vesicles. Elevated oxidative stress induced by dopamine oxidation may thus trigger the selective demise of DA neurons. Many strategies have been developed to protect DA neurons by stabilizing microtubules either directly or through intracellular signaling cascades. On the other hand, parkin, one of the most frequently mutated genes in PD, encodes for a protein-ubiquitin E3 ligase that strongly binds to microtubules. Parkin stabilizes microtubules through three domains that provide strong and independent interactions with tubulin and microtubules. These interactions anchor parkin on microtubules and may facilitate its E3 ligase activity on misfolded proteins transported along microtubules. Thus, parkin and rotenone, two prominent genetic and environmental factors linked to PD, act in an opposing manner on the same molecular target in the cell, microtubules, whose destruction underlies the selective vulnerability of dopaminergic neurons. PMID- 17079514 TI - The role of sleep in memory consolidation and brain plasticity: dream or reality? AB - The notion that a good night of sleep improves memory is widely accepted by the general public. Among sleep scientists, however, the idea has been hotly debated for decades. In this review, the authors consider current evidence for and against the hypothesis that sleep facilitates memory consolidation and promotes plastic changes in the brain. They find that despite a steady accumulation of positive findings over the past decade, the precise role of sleep in memory and brain plasticity remains elusive. This impasse may be resolved by more integrated approaches that combine behavioral and neurophysiological measurements in well described in vivo models of synaptic plasticity. PMID- 17079515 TI - Vicarious function of remote cortex following stroke: recent evidence from human and animal studies. AB - Following a lesion, the adult central nervous system undergoes dramatic structural and physiological reorganization in diverse subcortical and cortical areas. Our knowledge of the events that parallel recovery within the tissue surrounding the lesion and other distant cortical areas has evolved greatly in the past few years. Particularly, recent efforts have increased our understanding of the potential implication of premotor areas in recovery from lesions disturbing the primary motor cortex (M1) and its corticospinal outputs. Because these areas share extensive connections with M1 and have direct access to the spinal cord through corticospinal projections, they are particularly well positioned to take over the role of M1 in a vicarious manner and thus compensate for the neuronal loss resulting from M1 lesions. The impressive postlesional reorganization known to occur in many areas of the CNS including the premotor cortex traditionally has been assumed to play a beneficial role in recovery. However, recent experiments suggest that in some cases, reorganization of distant cortical areas correlates with poor recovery, raising the concept of maladaptive vicarious process. This concept might be particularly critical in the development of new treatment approaches favoring postlesion plasticity and even more so for interventions targeting specific area(s). Here, the author reviews human and animal studies that show the plastic potential of the adult CNS after stroke, highlighting the vicarious role of the premotor cortex in the recovery of motor control. PMID- 17079516 TI - Hedonic hot spots in the brain. AB - Hedonic "liking" for sensory pleasures is an important aspect of reward, and excessive 'liking' of particular rewards might contribute to excessive consumption and to disorders such as obesity. The present review aims to summarize recent advances in the identification of brain substrates for food 'liking' with a focus on opioid hot spots in the nucleus accumbens and ventral pallidum. Drug microinjection studies have shown that opioids in both areas amplify the 'liking' of sweet taste rewards. Modern neuroscience tools such as Fos plume mapping have further identified hedonic hot spots within the accumbens and pallidum, where opioids are especially tuned to magnify 'liking' of food rewards. Hedonic hot spots in different brain structures may interact with each other within the larger functional circuitry that interconnects them. Better understanding of how brain hedonic hot spots increase the positive affective impact of natural sensory pleasures will help characterize the neural mechanisms potentially involved in 'liking' for many rewards. PMID- 17079517 TI - Small-world brain networks. AB - Many complex networks have a small-world topology characterized by dense local clustering or cliquishness of connections between neighboring nodes yet a short path length between any (distant) pair of nodes due to the existence of relatively few long-range connections. This is an attractive model for the organization of brain anatomical and functional networks because a small-world topology can support both segregated/specialized and distributed/integrated information processing. Moreover, small-world networks are economical, tending to minimize wiring costs while supporting high dynamical complexity. The authors introduce some of the key mathematical concepts in graph theory required for small-world analysis and review how these methods have been applied to quantification of cortical connectivity matrices derived from anatomical tract tracing studies in the macaque monkey and the cat. The evolution of small-world networks is discussed in terms of a selection pressure to deliver cost-effective information-processing systems. The authors illustrate how these techniques and concepts are increasingly being applied to the analysis of human brain functional networks derived from electroencephalography/magnetoencephalography and fMRI experiments. Finally, the authors consider the relevance of small-world models for understanding the emergence of complex behaviors and the resilience of brain systems to pathological attack by disease or aberrant development. They conclude that small-world models provide a powerful and versatile approach to understanding the structure and function of human brain systems. PMID- 17079518 TI - Magnetoencephalography as a research tool in neuroscience: state of the art. AB - Magnetoencephalography (MEG) is a noninvasive neuroimaging method for detecting, analyzing, and interpreting the magnetic field generated by the electrical activity in the brain. Modern hardware can capture the MEG signal at hundreds of points around the head in a snapshot lasting only a fraction of a millisecond. The sensitivity of modern hardware is high enough to permit the extraction of a clean signal generated by the brain well above the noise level of the MEG hardware. It is possible to identify signatures of superficial and often deep generators in the raw MEG signal, even in snapshots of data. In a more quantitative way, tomographic images of the electrical current density in the brain can be extracted from each snapshot of MEG signal, providing a direct correlate of coherent collective neuronal activity. A number of recent studies have scrutinized brain function in the new spatiotemporal window that real-time tomographic analysis of MEG signals has opened. The results have allowed the variability in a single area to be seen in the context of activity in other areas and background rhythmic activity. In this view, normal brain function is seen as a cascade of extremely fast events and the unfolding of specialized processes, segregated in space and time and organized into well-defined stages of processing. PMID- 17079519 TI - Postdoctoral core competencies in the predoctoral curriculum. PMID- 17079520 TI - Interventions need to be consistent with osteopathic philosophy. PMID- 17079522 TI - Benign joint hypermobility syndrome: evaluation, diagnosis, and management. AB - Benign joint hypermobility syndrome (BJHS) is a connective tissue disorder with hypermobility in which musculoskeletal symptoms occur in the absence of systemic rheumatologic disease. Although BJHS has been well recognized in the rheumatology and orthopedic literature, it has not been discussed in the family medicine literature. Because most patients with musculoskeletal complaints are first seen by family physicians, it behooves primary care physicians to be familiar with recognizing and diagnosing BJHS. When patients with this syndrome are first seen by a physician, their chief complaint is joint pain, so BJHS can be easily overlooked and not considered in the differential diagnosis. Use of the Brighton criteria facilitates the diagnosis of BJHS. Treatment modalities include patient education, activity modification, stretching and strengthening exercises for the affected joint, and osteopathic manipulative treatment. PMID- 17079523 TI - Stretch reflex and Hoffmann reflex responses to osteopathic manipulative treatment in subjects with Achilles tendinitis. AB - CONTEXT: Irvin M. Korr, PhD, hypothesized that sensitivity of the monosynaptic stretch reflex (ie, deep tendon reflex) plays a major role in the restriction-of motion characteristic of somatic dysfunction, and that restoration of range of motion through osteopathic manipulative treatment (OMT) could be achieved by resetting of the stretch receptor gain. OBJECTIVE: To test Korr's hypothesis in the context of Achilles tendinitis, examining whether OMT applied to patients with Achilles tendinitis reduces the strength of the stretch reflex. METHODS: Subjects were recruited through public advertisements and referrals from healthcare professionals. There were no recruitment restrictions based on demographic factors. Amplitudes for stretch reflex and H-reflex (Hoffmann reflex) in the triceps surae muscles (the soleus together with the lateral and medial heads of the gastrocnemius) were measured in subjects with diagnosed Achilles tendonitis (n=16), both before and after OMT. These measurements were also made in asymptomatic control subjects (n=15) before and after sham manipulative treatment. RESULTS: As predicated on the concepts of the strain-counterstrain model developed by Lawrence H. Jones, DO, the use of OMT produced a 23.1% decrease in the amplitude of the stretch reflex of the soleus (P<.05) in subjects with Achilles tendinitis. Similarly significant responses were measured in the lateral and medial heads of the gastrocnemius in OMT subjects. The H-reflex was not significantly affected by OMT. In control subjects, neither reflex was significantly affected by sham manipulative treatment. By using a rating scale on questionnaires before treatment and daily for 7 days posttreatment, OMT subjects indicated significant clinical improvement in soreness, stiffness, and swelling. CONCLUSION: The reduction of stretch reflex amplitude with OMT, together with no change in H-reflex amplitude, is consistent with Korr's proprioceptive hypothesis for somatic dysfunction and patient treatment. Because subjects' soreness ratings also declined immediately after treatment, decreased nociceptor activity may play an additional role in somatic dysfunction, perhaps by altering stretch reflex amplitude. PMID- 17079524 TI - Effect of counterstrain on stretch reflexes, hoffmann reflexes, and clinical outcomes in subjects with plantar fasciitis. AB - CONTEXT: Previous research indicates that osteopathic manipulative treatment based on counterstrain produces a decrease in the stretch reflex of the calf muscles in subjects with Achilles tendinitis. OBJECTIVES: To study the effects of counterstrain on stretch reflex activity and clinical outcomes in subjects with plantar fasciitis. METHODS: In a single-blind, randomized controlled trial of crossover design, the effects of counterstrain were compared with those of placebo in adult subjects (N=20) with plantar fasciitis. The subjects were led to believe that both the counterstrain and placebo were therapeutic modalities whose effects were being compared. Ten subjects (50%) were assigned to receive 3 weeks of counterstrain treatment during phase 1 of the trial, while the other 10 subjects were given placebo capsules. After a 2- to 4-week washout period, phase 2 of the trial began with the interventions reversed. Clinical outcomes were assessed with daily questionnaires. Stretch reflex and H-reflex (Hoffmann reflex) in the calf muscles were assessed twice during each laboratory visit, before and after treatment in the counterstrain phase. RESULTS: No significant changes in the electrically recorded reflexes of the calf muscles were observed in response to treatment. However, changes in the mechanical characteristics of the twitches resulting from the electrical responses were observed. Peak force and time to reach peak force both increased (P< or =.05) in the posttreatment measurements, with the increase being significantly more pronounced in the counterstrain phase (P< .05). A comparison of pretreatment and posttreatment symptom severity demonstrated significant relief of symptoms that was most pronounced immediately following treatment and lasted for 48 hours. CONCLUSIONS: Clinical improvement occurs in subjects with plantar fasciitis in response to counterstrain treatment. The clinical response is accompanied by mechanical, but not electrical, changes in the reflex responses of the calf muscles. The causative relation between the mechanical changes and the clinical responses remains to be explored. PMID- 17079525 TI - Osteopathic medicine in transition: postmortem of the Osteopathic Medical Center of Texas. AB - The stand-alone osteopathic hospital was a necessity to the osteopathic medical profession in an era when it was isolated from allopathic medicine. As osteopathic medicine has become increasingly integrated with allopathic medicine, however, an independent osteopathic hospital is no longer a necessity. Moreover, a stand-alone institution seems to be economically out of place in today's market. The Osteopathic Medical Center of Texas in Fort Worth is an example of a stand-alone hospital that was unable to capitalize on the benefits realized by integrated hospital systems. The author believes that this failure contributed to the institution's demise. The market power of a hospital system can be used for more favorable contracting with vendors and providers, as well as facilitating negotiations with payers. System affiliation provides economic efficiency, security, and protection in the highly uncertain, complex, and competitive healthcare market. PMID- 17079526 TI - A safety review of long-acting beta2-agonists in patients with asthma. AB - Inhaled corticosteroids are the mainstay of asthma therapy; however, inhaled long acting beta2-agonists (LABAs) are frequently used in the treatment of patients with asthma. LABAs are combined with high-dose inhaled corticosteroids (ICSs) for patients with severe persistent asthma, and they are combined with low-dose ICSs for patients older than 5 years with moderate persistent asthma. Recent safety concerns raised by data from the Salmeterol Multi-Center Research Trial (SMART) have indicated that use of LABAs in some populations may contribute to increased mortality. These concerns are warranted when LABAs are used as monotherapy in the treatment of patients with asthma in whom they may cause increased exacerbations, blunting of rescue-medication effect, and worsening symptoms. However, when used in combination with an ICS, they decrease both rescue-medication use and symptoms, increase lung function, and act as steroid-sparing agents. PMID- 17079527 TI - How to predict USMLE scores from COMLEX-USA scores: a guide for directors of ACGME-accredited residency programs. AB - CONTEXT: Graduates of colleges of osteopathic medicine (COMs) frequently apply to residency training programs accredited by the Accreditation Council for Graduate Medical Education. However, students who have taken the Comprehensive Osteopathic Medical Licensing Examination (COMLEX-USA) rather than the United States Medical Licensing Examination (USMLE) may encounter a selection bias when applying to allopathic residency programs. OBJECTIVE: To evaluate the correlation between scores on COMLEX-USA and USMLE by COM students who took both examinations. METHODS: Scores were analyzed from a cohort of COM students who took COMLEX-USA Level 1 and USMLE Step 1 and a cohort of students who, during the same time, took COMLEX-USA Level 2 and USMLE Step 2. RESULTS: At the Kirksville (Mo) College of Osteopathic Medicine of A.T. Still University of Health Sciences, 155 students took both COMLEX-USA Level 1 and USMLE Step 1 and another cohort of 56 students took COMLEX-USA Level 2 and USMLE Step 2. The Pearson product moment correlation of the scores from COMLEX-USA Level 1 and USMLE Step 1 was 0.83 (P<.001) and for COMLEX-USA Level 2 and USMLE Step 2 was 0.70 (P<.001). Scores on USMLE are predicted by the corresponding COMLEX-USA scores using the equations USMLE Step 1 = 67.97 + 0.24 x COMLEX-USA Level 1 (R(2)=0.68); and USMLE Step 2 = 102.2 + 0.18 x COMLEX-USA Level 2 (R(2)=0.46). CONCLUSION: A method of estimating USMLE scores from COMLEX-USA scores is provided for residency directors of institutions accredited by the Accreditation Council for Graduate Medical Education for the evaluation of COM applicants. PMID- 17079528 TI - Computer-assisted instruction: a survey on the attitudes of osteopathic medical students. AB - OBJECTIVES: To investigate the views of osteopathic medical students on their preferred learning methods, their current use of computers as an educational tool, and their attitudes regarding the role of computers in medical education, based on their skill level. METHODS: A 27-item questionnaire was distributed to first-through fourth-year osteopathic medical students. Items asked students to assess their levels of computer skills and experience, their current use of computers as an educational tool, and their attitudes regarding the role of computers in medical education. RESULTS: Of the 246 students (80% of enrolled students) who responded to the questionnaire, a majority (129 [53%]) rated themselves as having intermediate computer skills, and the remaining students rated their skills as basic (69 [28%]) or advanced (46 [19%]). Most students (176 [72%]) felt that they learned best by both hearing and seeing or reading new material, that they learned more easily from a printed page than a computer screen, and that live lectures provided them with the best opportunity for learning. However, when compared with students who have basic and intermediate computer skills, students with advanced computer skills were more in favor of computer-assisted instruction and testing as well as a school requirement to own a computer and to use a laptop computer in class. CONCLUSIONS: Students' opinions of the importance of computer technology in medical education seem to be based mainly on their self-assessed technical competency levels. Understanding this dynamic may aid medical educators in the implementation of computer-assisted instruction. PMID- 17079529 TI - Association between health care providers' influence on parents who have concerns about vaccine safety and vaccination coverage. AB - OBJECTIVES: Parents who have concerns about vaccine safety may be reluctant to have their children vaccinated. The purpose of this study was to explore how vaccination coverage among children 19 to 35 months of age is associated with health care providers' influence on parents' decision to vaccinate their children, and with parents' beliefs about vaccine safety. METHODS: Parents of 7695 children 19 to 35 months of age sampled by the National Immunization Survey were administered the National Immunization Survey Parental Knowledge Module between the third quarter of 2001 and the fourth quarter of 2002. Health care providers were defined as a physician, nurse, or any other type of health care professional. Parents provided responses that summarized the degree to which they believed vaccines were safe, and the influence providers had on their decisions to vaccinate their children. Children were determined to be up-to-date if their vaccination providers reported administering > or = 4 doses of diphtheria and tetanus toxoids and acellular pertussis vaccine, > or = 3 doses of polio vaccine, > or = 1 dose of measles-mumps-rubella vaccine, > or = 3 doses of Haemophilus influenzae type b vaccine, and > or = 3 doses of hepatitis B vaccine. RESULTS: Of all of the parents, 5.7% thought that vaccines were not safe, and 21.5% said that their decision to vaccinate their children was not influenced by a health care provider. Compared with parents who responded that providers were not influential in their decision to vaccinate their children, parents who responded that providers were influential were twice as likely to respond that vaccines were safe for children. Among children whose parents believed that vaccines were not safe, those whose parents' decision to vaccinate was influenced by a health care provider had an estimated vaccination coverage rate that was significantly higher than the estimated coverage rate among children whose parents' decision was not influenced by a health care provider (74.4% vs 50.3%; estimated difference: 24.1%). CONCLUSIONS: Health care providers have a positive influence on parents to vaccinate their children, including parents who believe that vaccinations are unsafe. Physicians, nurses, and other health care professionals should increase their efforts to build honest and respectful relationships with parents, especially when parents express concerns about vaccine safety or have misconceptions about the benefits and risks of vaccinations. PMID- 17079530 TI - Heat or eat: the Low Income Home Energy Assistance Program and nutritional and health risks among children less than 3 years of age. AB - OBJECTIVES: Public funding for the Low Income Home Energy Assistance Program has never been sufficient to serve more than a small minority of income-eligible households. Low Income Home Energy Assistance Program funding has not increased with recent rapidly rising energy costs, harsh winter conditions, or higher child poverty rates. Although a national performance goal for the Low Income Home Energy Assistance Program is to increase the percentage of recipient households having > or = 1 member < or = 5 years of age, the association of income-eligible households' receipt of the Low Income Home Energy Assistance Program with indicators of well-being in young children has not been evaluated previously. The goal of the current study was to evaluate the association between a family's participation or nonparticipation in the Low Income Home Energy Assistance Program and the anthropometric status and health of their young children. METHODS: In the ongoing Children's Sentinel Nutrition Assessment Project from June 1998 through December 2004, caregivers with children < 3 years of age in 2 emergency departments and 3 primary care clinics in 5 urban sites participated in cross-sectional surveys regarding household demographics, child's lifetime history of hospitalizations, and, for the past 12 months, household public assistance program participation and household food insecurity, measured by the US Food Security Scale. This scale, in accordance with established procedures, classifies households as food insecure if they report that they cannot afford enough nutritious food for all of the members to lead active, healthy lives. On the day of the interview, children's weight, length, and whether the children were admitted acutely to the hospital from the emergency departments were documented. The study sample consisted only of Low Income Home Energy Assistance Program income-eligible renter households without private insurance who also participated in > or = 1 other means-tested program. RESULTS: In this sample of 7074 caregivers, 16% of families received the Low Income Home Energy Assistance Program, similar to the national rate of 17%. Caregivers who received the Low Income Home Energy Assistance Program were more likely to be single (63% vs 54%), US born (77% vs 68%), and older (mother's mean age: 28.1 vs 26.7 years) but were less likely to be employed (44% vs 47%). Households who received the Low Income Home Energy Assistance Program were more likely to receive Supplemental Nutrition Program for Women, Infants, and Children (85% vs 80%), Supplemental Security Income (13% vs 9%), Temporary Assistance for Needy Families (38% vs 23%), and food stamps (59% vs 37%) and to live in subsidized housing (38% vs 19%) compared with nonrecipients. Children in families participating in the Low Income Home Energy Assistance Program were older than children in nonparticipating families (13.6 vs 12.5 months), were less likely to be uninsured (5% vs 9%), and were more likely to have had a low birth weight < or = 2500 g (17% vs 14%). Families participating in the Low Income Home Energy Assistance Program reported more household food insecurity (24% vs 20%) There were no significant group differences between recipients and nonrecipients in caregiver's education or child's gender. After controlling for these potentially confounding variables, including receipt of other means-tested programs, compared with children in recipient households, those in nonrecipient households had greater adjusted odds of being at aggregate nutritional risk for growth problems, defined as children with weight-for-age below the 5th percentile or weight-for-height below the 10th percentile, with significantly lower mean weight-for-age z scores calculated from age- and gender-specific values from the Centers for Disease Control and Prevention 2000 reference data. However, in adjusted analyses, children aged 2 to 3 years in recipient households were not more likely to be overweight (BMI > 95th percentile) than those in nonrecipient households. Rates of age-adjusted lifetime hospitalization excluding birth and the day of the interview did not differ between Low Income Home Energy Assistance Program recipient groups. Among the 4445 of 7074 children evaluated in the 2 emergency departments, children from eligible households not receiving the Low Income Home Energy Assistance Program had greater adjusted odds than those in recipient households of acute hospital admission on the day of the interview. CONCLUSIONS: Even within a low-income renter sample, Low Income Home Energy Assistance Program benefits seem to reach families at the highest social and medical risk with more food insecurity and higher rates of low birth-weight children. Nevertheless, after adjustment for differences in background risk, living in a household receiving the Low Income Home Energy Assistance Program is associated with less anthropometric evidence of undernutrition, no evidence of increased overweight, and lower odds of acute hospitalization from an emergency department visit among young children in low income renter households compared with children in comparable households not receiving the Low Income Home Energy Assistance Program. The Low Income Home Energy Assistance Program in many states shuts down early each winter when their funding is exhausted. From a clinical perspective, pediatric health providers caring for children from impoverished families should consider encouraging families of these children to apply for the Low Income Home Energy Assistance Program early in the season before funding is depleted. From a public policy perspective, although this cross-sectional study design can only demonstrate associations and not causation, these findings suggest that, particularly as fuel costs and children's poverty rates increase, expanding the Low Income Home Energy Assistance Program funding and meeting the national Low Income Home Energy Assistance Program performance goal of increasing the percentage of recipient households with young children might potentially benefit such children's growth and health. PMID- 17079531 TI - Reducing children's television-viewing time: a qualitative study of parents and their children. AB - OBJECTIVES: The American Academy of Pediatrics recommends that children over age 2 years spend < or = 2 hours per day with screen media, because excessive viewing has been linked to a plethora of physical, academic, and behavioral problems. The primary goal of this study was to qualitatively explore how a recommendation to limit television viewing might be received and responded to by a diverse sample of parents and their school-age children. METHODS: The study collected background data about media use, gathered a household media inventory, and conducted in depth individual and small group interviews with 180 parents and children ages 6 to 13 years old. RESULTS: Most of the children reported spending approximately 3 hours per day watching television. The average home in this sample had 4 television sets; nearly two thirds had a television in the child's bedroom, and nearly half had a television set in the kitchen or dining room. Although virtually all of the parents reported having guidelines for children's television viewing, few had rules restricting the time children spend watching television. Data from this exploratory study suggest several potential barriers to implementing a 2-hour limit, including: parents' need to use television as a safe and affordable distraction, parents' own heavy television viewing patterns, the role that television plays in the family's day-to-day routine, and a belief that children should spend their weekend leisure time as they wish. Interviews revealed that for many of these families there is a lack of concern that television viewing is a problem for their child, and there remains confusion about the boundaries of the recommendation of the American Academy of Pediatrics. CONCLUSIONS: Parents in this study expressed interest in taking steps toward reducing children's television time but also uncertainty about how to go about doing so. Results suggest possible strategies to reduce the amount of time children spend in front of the screen. PMID- 17079532 TI - School performance and social-emotional behavior of primary school children before and after a disaster. AB - OBJECTIVES: The purpose of this work was to evaluate the cognitive and social emotional consequences in a general population of primary school children affected by the firework disaster in Enschede, The Netherlands, on May 13, 2000. The explosions caused tremendous damage in the surrounding neighborhood. Twenty two people immediately died and >1000 were injured. METHODS: This retrospective study assessed school performance and social-emotional behavior before and up to 3 years after the disaster. Objectively measured school test results in spelling and arithmetic/mathematics and multi-informant social-emotional behavioral assessments were compared between exposed and nonexposed primary school children. Multivariate logistic-regression was used to assess the relationship between exposure and cognitive and social emotional functioning. RESULTS: On school performances, the children exposed to the disaster performed over a period of 3 years after the disaster as good as or better than classmates, controls, and a national reference population. Shortly after the disaster, exposed children even seemed to have better school test results than nonexposed children. Two to 3 years after the disaster, a significant effect of disaster exposure was found on social-emotional behavior. Problematic behavior was reported by teachers, parents, and the school doctor. CONCLUSIONS: This study demonstrates a limited influence of disaster exposure on school performance in primary school children. This study also shows that teachers and youth health care practitioners especially should be aware of children starting school several years after a disaster. Although very young at the time of a disaster (1-4 years of age), they may experience disaster-related problems. PMID- 17079533 TI - Direct medical cost of influenza-related hospitalizations in children. AB - OBJECTIVE: Our goal was to determine the cost of influenza-related hospitalization in children with community-acquired laboratory-confirmed influenza and to identify predictors of high hospitalization costs. PATIENTS AND METHODS: This was a retrospective cohort study of patients 21 years and younger hospitalized at a children's hospital with community-acquired laboratory confirmed influenza during 4 consecutive influenza seasons (2000-2004). The main outcome measure was the direct medical cost of influenza-related hospitalizations, including the cost of diagnostics, therapeutics, room, and physician services. RESULTS: Electronic billing data were retrievable for 727 (98%) of 745 patients hospitalized for community-acquired laboratory-confirmed influenza during the study period. A total of 478 (66%) children were in a high risk group for whom the Advisory Committee on Immunization Practices recommended influenza vaccine (patients with Advisory Committee on Immunization Practices designated chronic medical conditions or aged 6-23 months). The mean total cost of hospitalization for influenza-related illness was 13,159 dollars (39,792 dollars for patients admitted to an ICU; 7030 dollars for patients cared for exclusively on the wards). High-risk patients had higher mean total costs (15,269 dollars) than low-risk patients (9107 dollars). Cardiac, metabolic, and neurologic/neuromuscular diseases and age of 18 to 21 years were independently associated with the highest hospitalization costs (>15th percentile). CONCLUSIONS: The cost of influenza-related hospitalizations in children may be considerably higher than previously estimated. The presence of certain Advisory Committee on Immunization Practices-designated chronic medical conditions is associated with higher influenza-related hospitalization costs. Successfully immunizing patients with these conditions has the potential for significant cost savings. PMID- 17079534 TI - Changes in the use of postnatal steroids for bronchopulmonary dysplasia in 3 large neonatal networks. AB - BACKGROUND: Postnatal corticosteroids were widely used in the 1990s in an attempt to reduce the incidence of bronchopulmonary dysplasia. However, high rates of short-term adverse effects and impaired neurodevelopmental outcomes were seen. In early 2002, a joint statement of the American Academy of Pediatrics and Canadian Paediatric Society called for limitation in the use of postnatal corticosteroids. The impact of this statement is not known. OBJECTIVES: The purpose of this work was to determine the frequency of postnatal corticosteroid use and mortality and morbidities over time, particularly before and after the joint statement. DESIGN/METHODS: We conducted a retrospective analysis of cohort data within 3 large network registries (the National Institute of Child Health and Development Neonatal Research Network [18 centers], the Vermont Oxford Network [444 centers], and the Canadian Neonatal Network [10 centers]) for the following 3 periods: prestatement (2001), statement (2002), and poststatement (2003) of very low birth weight infants (501-1500 g). The National Institute of Child Health and Development Neonatal Research Network and the Vermont Oxford Network were also analyzed for longer-term trends from 1990 to 2003. Postnatal corticosteroid use, mortality at discharge, and neonatal morbidities (bronchopulmonary dysplasia at 36 weeks, late-onset infection >72 hours of age, necrotizing enterocolitis treated with surgery, and length of stay) between periods were compared. RESULTS: Mean birth weight (range: 1022-1060 g), postmenstrual age (28 weeks), and gender (51% male) were similar between the networks. Race differed with more black infants in the National Institute of Child Health and Development Neonatal Research Network than the Vermont Oxford Network (38% vs 24%). Antenatal steroid use was similar (range: 61%-75%). Postnatal corticosteroid use rose from 1990 (8% 16%), peaked in 1996-1998 (24%-28%), and began to decline in 1999. Use in 2003 was significantly less than in 2001. Mortality and major morbidities were similar. CONCLUSIONS: Postnatal corticosteroid use had decreased significantly in 3 large neonatal networks before the joint statement with further decreases after the statement with no apparent impact on mortality and short-term morbidity. Despite substantial decreases, approximately 8% of very low birth-weight infants continue to be treated with postnatal corticosteroid. PMID- 17079535 TI - Illness and injury among children attending summer camp in the United States, 2005. AB - OBJECTIVES: The purpose of this work was to describe illnesses and injuries sustained by campers at summer camps, calculate illness and injury rates, and identify risk factors for camp-related illness and injury. METHODS: A convenience sample of camps selected from the US summer camp population participated in this pilot illness and injury surveillance study. Camps completed weekly reports detailing the number of campers on site and the number of adverse events sustained and provided specific information about each adverse event. RESULTS: A total of 177 camper illnesses and injuries occurred during 122,379 camper-days, with a median rate of 1.15 adverse events per 1000 camper-days. The majority of reported events were illnesses (68.0%), 11.8% of which were communicable and seen in multiple individuals at camp. Of the injury events (32.0%), cut/scratch/scrape were the most common diagnoses (33.3%), followed by fracture (14.6%) and sprain/strain (10.4%); horseback riding and capture the flag were identified as injury-producing activities. CONCLUSIONS: Internet- and telephone-based surveillance systems can be successfully used to conduct illness and injury surveillance among children attending summer camp. Data collected via such systems can be used to calculate illness and injury rates, to describe patterns of illness and injury, and to identify risk factors for camper-related illness and injury. Given the millions of children attending summer camp yearly, a surveillance system such as this can provide the data needed to develop evidence based prevention interventions to decrease the number of youth whose camp experiences are negatively affected by illness and injury. PMID- 17079536 TI - A refined symptom-based approach to diagnose pulmonary tuberculosis in children. AB - BACKGROUND: Tuberculosis control programs place an almost exclusive emphasis on adults with sputum smear-positive tuberculosis, because they are most infectious. However, children contribute a significant proportion of the global tuberculosis caseload and experience considerable tuberculosis-related morbidity and mortality, but few children in endemic areas have access to antituberculosis treatment. The diagnostic difficulty experienced in endemic areas with limited resources has been identified as a major factor contributing to poor treatment access. In general, there is a sense of scepticism regarding the potential value of symptom-based diagnostic approaches, because current clinical diagnostic approaches are often poorly validated. The natural history of childhood tuberculosis demonstrates that symptoms may offer good diagnostic value if they are well defined and if appropriate risk stratification is applied. This study aimed to determine the value of well-defined symptoms to diagnose childhood pulmonary tuberculosis in a tuberculosis-endemic area. METHODS: A prospective, community-based study was conducted in Cape Town, South Africa. Specific well defined symptoms were documented in all children < 13 years of age reporting a persistent, nonremitting cough of > 2 weeks' duration; study participants were thoroughly evaluated for tuberculosis. In addition, all of the children who received antituberculosis treatment during the study period were reviewed by the investigator, irrespective of study inclusion. This concurrent disease surveillance provided a comprehensive overview of all of the childhood tuberculosis cases, allowing accurate assessment of the possible disadvantages associated with this symptom-based diagnostic approach. In the absence of an acceptable gold standard test, optimal case definition is an important consideration. Children were categorized as "bacteriologically confirmed tuberculosis," "radiologically certain tuberculosis," "probable tuberculosis," or "not tuberculosis." Bacteriologically confirmed tuberculosis was defined as the presence of acid-fast bacilli on sputum microscopy and/or Mycobacterium tuberculosis cultured from a respiratory specimen. Radiologically certain tuberculosis was defined as agreement between both independent experts that the chest radiograph indicated certain tuberculosis in the absence of bacteriologic confirmation. Probable tuberculosis was defined as the presence of suggestive radiologic signs and good clinical response to antituberculosis treatment in the absence of bacteriologic confirmation or radiologic certainty. Good clinical response was defined as complete symptom resolution and weight gain of > or = 10% of body weight at diagnosis, within 3 months of starting antituberculosis treatment. Not tuberculosis was defined as spontaneous symptom resolution or no response to antituberculosis therapy in the absence of bacteriologic confirmation or radiologic signs suggestive of tuberculosis. Pulmonary tuberculosis was defined as a symptomatic child with: (1) bacteriologically confirmed tuberculosis, (2) radiologically confirmed tuberculosis, or (3) probable tuberculosis (as defined), excluding isolated pleural effusion. RESULTS: In total, 1024 children were referred for evaluation. Resolving symptoms were reported in 596 children (58.2%); 428 (41.8%) children with persistent, nonremitting symptoms at evaluation were investigated for tuberculosis. Pulmonary tuberculosis was diagnosed in 197 children; 96 were categorized as bacteriologically confirmed tuberculosis, 75 as radiologically certain tuberculosis, and 26 as probable tuberculosis. Combining a persistent nonremitting cough of > 2 weeks' duration, documented failure to thrive (in the preceding 3 months), and fatigue provided reasonable diagnostic accuracy in HIV uninfected children (sensitivity: 62.6%; specificity: 89.8%; positive predictive value: 83.6%); the performance was better in the low-risk group (> or = 3 years; sensitivity: 82.3%; specificity: 90.2%; positive predictive value: 82.3%) than in the high-risk group (< 3 years; sensitivity: 51.8%; specificity: 92.5%; positive predictive value: 90.1%). In children with an uncertain diagnosis at presentation, clinical follow-up was a valuable diagnostic tool that further improved diagnostic accuracy, particularly in the low-risk group. Symptom-based approaches offered little diagnostic value in HIV-infected children. Three (15%) of the 20 HIV-infected children diagnosed with pulmonary tuberculosis failed to report symptoms of sufficient duration to warrant study inclusion, whereas 25% reported persistent, nonremitting symptoms in the absence of tuberculosis. In addition, the tuberculin skin test was positive in < 20% of HIV-infected children diagnosed with pulmonary tuberculosis. DISCUSSION: The combined presence of 3 well-defined symptoms at presentation (persistent, nonremitting cough of > 2 weeks' duration; objective weight loss [documented failure to thrive] during the preceding 3 months; and reported fatigue) provided good diagnostic accuracy in HIV-uninfected children > or = 3 years of age, with clinical follow-up providing additional value. The approach performed less well in children < 3 years. However, the presence of a persistent, nonremitting cough together with documented failure to thrive still provided a fairly accurate diagnosis (sensitivity: 68.3%; specificity: 80.1%; positive predictive value: 82.1%), illustrating the importance of regular weight monitoring in young children. Clinical follow-up also offered additional diagnostic value, but caution is required, because very young children have an increased risk of rapid disease progression. The approach performed poorly in HIV-infected children. Recent household contact with an adult index case seemed to provide more diagnostic value than a positive tuberculin skin test, but novel T-cell-based assays may offer the only real improvement in sensitivity to diagnose M. tuberculosis infection in HIV-infected children. The variable diagnostic value offered by this symptom-based diagnostic approach illustrates the importance of risk stratification, as demonstrated by the fact that 11 (91.7%) of 12 children with severe disease manifestations who failed to meet the entry criteria were < 3 years of age or HIV infected. Particular emphasis should be placed on the provision of preventive chemotherapy after documented exposure and/or infection in these high-risk children. Study limitations include the small number of HIV infected children, but on the positive side, the large number of HIV-uninfected children permitted adequate evaluation in this important group. It is often forgotten that HIV-uninfected children constitute the majority of child tuberculosis cases, even in settings where HIV is endemic. This study demonstrates the importance of ascertaining a child's HIV status before symptom based diagnosis is attempted. Because children were recruited at both the clinic and hospital level, some selection bias may have been introduced; however, the only significant difference between the 2 groups was the proportion of HIV infected children. Pulmonary tuberculosis was diagnosed with different levels of certainty, but no significant differences were recorded between these groups. CONCLUSIONS: Pulmonary tuberculosis can be diagnosed with a reasonable degree of accuracy in HIV-uninfected children using a simple symptom-based approach. This offers the exciting prospect of improving treatment access for children, particularly in resource-limited settings where current access to antituberculosis treatment is poor. PMID- 17079537 TI - Atopic disposition, wheezing, and subsequent respiratory syncytial virus hospitalization in Danish children younger than 18 months: a nested case-control study. AB - OBJECTIVES: This study examined whether atopic disposition, wheezing, and atopic disorders increased the risk of hospitalizations because of respiratory syncytial virus in children between birth and 18 months of age. PATIENTS AND METHODS: Relative risks for respiratory syncytial virus hospitalization were studied in a nested 1:5 case-control design using exposure information obtained from interviews with mothers of 2564 case and 12,816 control children who had been followed prospectively from birth and until 18 months of age as participants in the Danish National Birth Cohort. Information on the children's ages at respiratory syncytial virus hospitalization, presentation of infrequent wheezing, recurrent wheezing, and atopic dermatitis were used to study these associations chronologically. RESULTS: The adjusted relative risk of respiratory syncytial virus hospitalization in the offspring was 1.11 for maternal atopic dermatitis, 1.72 for maternal asthma, and 1.23 for paternal asthma. Atopic dermatitis in the child was associated with an increased risk of subsequent respiratory syncytial virus hospitalization among infants <6 months of age. Infrequent wheezing was associated with a relative risk of subsequent respiratory syncytial virus hospitalization of 2.98 and recurrent wheezing with a relative risk of 5.90. These associations were present also if infants with medical risk factors were excluded from the analysis. CONCLUSIONS: Asthmatic disposition and wheezing were strong determinants of subsequent respiratory syncytial virus hospitalization in Danish children <18 months of age. PMID- 17079538 TI - Compliance with American Academy of Pediatrics and American Public Health Association illness exclusion guidelines for child care centers in Maryland: who follows them and when? AB - BACKGROUND: In 1992, the American Academy of Pediatrics and the American Public Health Association jointly published guidelines for temporary exclusion of sick children from child care. However, little is known about key stakeholders' compliance with these guidelines. OBJECTIVES: The purpose of this work was to compare pediatricians', parents', and child care providers' compliance with American Academy of Pediatrics guidelines and determine predictors for higher rates of compliance. METHODS: We conducted a cross-sectional survey of 215 randomly selected Maryland pediatricians, 223 parents, and 192 child care providers from 22 Baltimore, Maryland, child care centers from January to July 2004. Questionnaires contained the following 6 case vignettes depicting common child care illnesses: upper respiratory infection, conjunctivitis, gastroenteritis, mild febrile illness, tinea capitis, and atopic dermatitis. The instrument measured the correctness of exclusion and inclusion decisions (using American Academy of Pediatrics/American Public Health Association guidelines as gold standard) according to varying levels of fever, disease severity (eg, clear versus yellow eye discharge), familiarity with the child, and parent work schedule flexibility. RESULTS: Response rates were 71% for pediatricians, 56% for parents, and 85% for child care providers. Guideline compliance was higher for pediatricians (74%) than for child care providers (60%) and parents (61%). Only 23% of pediatricians and parents and 29% of child care providers reported familiarity with American Academy of Pediatrics/American Public Health Association guidelines by name. In general, child care providers and parents had lower false-negative rates (allowed fewer children to attend who met criteria for exclusion) than pediatricians, suggesting that pediatricians may underexclude. Child care providers and parents correctly excluded in 65%-98% of cases requiring exclusion, whereas pediatricians correctly excluded 31%-86% of cases requiring exclusion, depending on the vignette. Yet pediatricians were much more specific about which children met criteria (pediatricians correctly included 61%-93% of cases requiring inclusion versus child care providers and parents who correctly included 20%-75% of such cases), suggesting that child care providers and parents may overexclude. Compliance rates varied significantly by stakeholder, vignette (disease), level of fever, and disease severity but did not vary with the stakeholder's familiarity with the child or the flexibility of the parent's work schedule. CONCLUSIONS: Pediatricians, parents, and child care providers were unfamiliar with American Academy of Pediatrics/American Public Health Association illness exclusion guidelines by name but moderately compliant with them. When noncompliant, child care providers and parents generally overexcluded, and pediatricians underexcluded. Stakeholder- and disease-specific predictors for noncompliance gleaned from this study suggest how educational interventions aiming to increase guideline compliance could be individually tailored to child care providers, parents, and pediatricians. PMID- 17079539 TI - Public parks and physical activity among adolescent girls. AB - OBJECTIVES: Physical activity may be constrained or facilitated by local environments. The availability of neighborhood facilities for physical activity may be particularly relevant for youth, who are unable to drive and whose activity is often limited to the immediate distance they are able to walk or bicycle. Several studies have shown that proximity to recreational facilities and parks is one of the most important predictors of physical activity. Because the United States already has an extensive infrastructure of parks, with 70% of adults indicating that they live within walking distance of a park or playground, parks may be a potential venue for increasing physical activity. This might be particularly important for adolescent girls, whose physical activity levels decline substantially as they go through puberty. The goal of this study was to examine the association between park proximity, park type, and park features and physical activity in adolescent girls. PATIENTS AND METHODS: This was a cross sectional study using baseline data from the Trial of Activity for Adolescent Girls. It included 1556 grade 6 girls who were randomly selected from 6 middle schools in each of the following 6 field site areas: Washington, DC, and Baltimore, Maryland; Columbia, South Carolina; Minneapolis, Minnesota; New Orleans, Louisiana; Tucson, Arizona; and San Diego, California. Girls wore accelerometers for 6 days to measure metabolic equivalent-weighted moderate-to vigorous physical activity, a measure accounting for the volume and intensity of activity. Metabolic equivalent-weighted moderate-to-vigorous physical activity was calculated for the hours outside of school time using 2 different cutpoints, activity levels > or = 3.0 metabolic equivalents and > or = 4.6 metabolic equivalents, the latter indicating activity at the intensity of a brisk walk or higher. We mapped all of the parks within 1 mile of each girl's home. Trained staff used a checklist to document the presence of facilities and amenities at each park, including passive amenities, such as drinking fountains, restrooms, and areas with shade, as well as active amenities like basketball courts, multipurpose fields, playgrounds, and tennis courts. RESULTS: Mean nonschool metabolic equivalent-weighted moderate/vigorous physical activity, using the 4.6 metabolic equivalent cutoff, was 611.1 minutes (range: 49.7-4718.6 metabolic equivalent minutes per 6 days) and 1704.8 metabolic equivalent minutes per 6 days (range: 276.2-5792.6 metabolic equivalent minutes per 6 days) when using the 3.0 metabolic equivalent cutpoint. Many girls had multiple parks within a 1-mile radius of their homes: 57% had > or = 1 type of park, the majority being neighborhood or community parks; 42% had between 1 and 3 parks, 37% had > or = 4 parks, and 14% had > or = 8 parks. The type, number, and specific parks features were associated with girls' nonschool metabolic equivalent-weighted moderate/vigorous physical activity. At the 4.6 metabolic equivalent cutpoint, higher levels of nonschool metabolic equivalent-weighted moderate/vigorous physical activity were associated with both neighborhood and community parks (22 metabolic equivalent minutes) and miniparks (40 metabolic equivalent minutes). Each park, regardless of type, in the half-mile around each girl's home was associated with an increase in nonschool metabolic equivalent-weighted moderate/vigorous physical activity by 2.8% or 17.2 nonschool minutes of metabolic equivalent-weighted moderate/vigorous physical activity per 6 days. Beyond a half-mile, each park increased nonschool metabolic equivalent-weighted moderate/vigorous physical activity by 1.1% or 6.7 metabolic equivalent minutes per 6 days. For the average girl with 3.5 parks within a 1-mile radius of home, the presence of parks accounted for 36.5 extra nonschool metabolic equivalent minutes per 6 days, approximately 6% of total nonschool metabolic equivalent weighted moderate/vigorous physical activity. Using the 3.0 metabolic equivalent cutpoint, this sums to an additional 68 metabolic equivalent minutes of nonschool metabolic equivalent-weighted moderate/vigorous physical activity over 6 days, or 4% of the total. The most common amenities in the parks were playgrounds, multipurpose fields, and picnic areas. Slightly more than one third of girls lived within a half-mile of a park with a basketball court, and > 20% had access to walking paths and tennis courts in their local park. Higher levels of nonschool metabolic equivalent-weighted moderate/vigorous physical activity per 6 days were associated with walking paths (13 metabolic equivalent minutes), running tracks (82 metabolic equivalent minutes), playgrounds (28 metabolic equivalent minutes), and basketball courts (30 metabolic equivalent minutes). Parks with streetlights and floodlights were also associated with an increase of 18 and 22 minutes of nonschool metabolic equivalent-weighted moderate/vigorous physical activity, respectively. With the 3.0 metabolic equivalent cutoff for metabolic equivalent-weighted moderate/vigorous physical activity, additional nonschool metabolic equivalent minutes more than doubled when girls had miniparks (92 metabolic equivalent minutes), natural resource areas (36 metabolic equivalent minutes), walking paths (59 metabolic equivalent minutes), and running tracks (208 metabolic equivalent minutes) within a half-mile of their homes. Skateboard areas and special-use parks were negatively associated with nonschool metabolic equivalent-weighted moderate/vigorous physical activity in adolescent girls. CONCLUSIONS: Adolescent girls who live near more parks, particularly near those with amenities that are conducive to walking and with active features, engage in more nonschool metabolic equivalent-weighted moderate/vigorous physical activity than those with fewer parks. Whether this is because of actual use of the parks or neighborhood choice could not be determined. Although the magnitude of the association between parks and additional minutes of metabolic equivalent weighted moderate/vigorous physical activity was small for an individual, amounting to an average of 4%-6% of a girl's total nonschool metabolic equivalent weighted moderate/vigorous physical activity, it is likely to have a large population-level association. Because of the potential population level impact, the use of parks to promote physical activity should be further studied. PMID- 17079540 TI - Recent trends in waist circumference and waist-height ratio among US children and adolescents. AB - OBJECTIVES: Abdominal obesity may be a better predictor than overall obesity for the risk of cardiovascular disease and type 2 diabetes. Waist circumference and waist-height ratio are 2 simple, yet effective, surrogate measures of abdominal obesity. We sought to examine the recent trends in mean waist circumference and waist-height ratio and prevalence of abdominal obesity among children and adolescents aged 2 to 19 years in the United States. METHODS: Representative samples of the civilian, noninstitutionalized US population from the National Health and Nutrition Examination Survey conducted during 4 time periods, 1988 1994 (ie, National Health and Nutrition Examination Survey III), 1999-2000, 2001 2002, and 2003-2004, were examined to estimate the mean waist circumference and waist-height ratio of boys and girls in 4 different age groups. Data from the 3 most recent National Health and Nutrition Examination Surveys were combined to establish a National Health and Nutrition Examination Survey 1999-2004 category. RESULTS: Categorized by age group, the unadjusted mean waist circumference for boys increased between National Health and Nutrition Examination Survey III and National Health and Nutrition Examination Survey 1999-2004 from 50.7 cm (aged 2-5 years), 61.9 cm (aged 6-11 years), 76.8 cm (aged 12-17 years), and 81.3 cm (aged 18-19 years) to 51.9, 64.5, 79.8, and 86.6 cm, respectively. During the same time periods and within the same age groups, the unadjusted mean waist circumference for girls increased from 51.0, 61.7, 75.0, and 77.7 cm to 51.8, 64.7, 78.9, and 83.9 cm, respectively. The relative change in waist-height ratio was similar to waist circumference at each age group for both boys and girls. Using the 90th percentile values of waist circumference for gender and age, the prevalence of abdominal obesity increased by 65.4% (from 10.5% to 17.4%) and 69.4% (from 10.5% to 17.8%) for boys and girls, respectively. CONCLUSIONS: Mean waist circumference and waist-height ratio and the prevalence of abdominal obesity among US children and adolescents greatly increased between 1988-1994 and 1999-2004. PMID- 17079541 TI - Variations in low birth weight and preterm delivery among blacks in relation to ancestry and nativity: New York City, 1998-2002. AB - OBJECTIVES: Black women in the United States are more likely to give birth to preterm and low birth-weight infants than their white counterparts, but little is known about variation in birth outcomes within the black population. This study aimed to test the hypothesis that the risk of low birth weight and preterm birth within the black population varies by maternal ancestry and nativity. POPULATION AND METHODS: We conducted a retrospective cohort study using New York City birth records. All of the recorded live births to black women occurring in New York City between January 1, 1998, and December 31, 2002 (N = 168,039), were divided into the following self-reported ancestry groups: African, American, Asian, Cuban, European, Puerto Rican, South and Central American (excluding Brazilian), and West Indian and Brazilian. To estimate adjusted risk ratios for low birth weight (weight at birth <2500 g) and preterm birth (gestational age at delivery <37 weeks, based on clinical estimate), we ran 3 models for each outcome, using negative binomial regression and Poisson regression with robust SE estimation. All of the models used blacks reporting American ancestry as the reference group. The first model included ancestry as the primary exposure variable along with covariates that included maternal age, parity, smoking, and education, as well as paternal education and race. Nativity (US- or foreign-born) was included in the second model, and terms representing interaction effects between ancestry and nativity were included in the third model. RESULTS: There was substantial variation in risks of preterm birth and low birth weight among the black subgroups, with all of the groups having lower risks than the American black reference group, even after adjusting for maternal risk factors and other covariates. Risk ratios for low birth weight ranged from 0.55 among South/Central Americans to 0.91 among Cubans; risk ratios for preterm birth showed a similar pattern. Nativity was also associated with low birth weight and preterm birth; births to foreign-born women were less likely to be preterm or low birth weight than births to US-born women. Furthermore, nativity effects varied by ancestry group, with foreign-born status inversely associated with poor birth outcomes among South/Central Americans but not among West Indians/Brazilians. CONCLUSIONS: Important health differences may be masked in studies that treat black women in America as a homogeneous group and do not take ethnic variation and nativity into account. PMID- 17079542 TI - The association of child and household food insecurity with childhood overweight status. AB - CONTEXT: The prevalence of childhood overweight status is increasing. Some have suggested that childhood overweight is associated with food insecurity, defined as limited or uncertain access to enough nutritious food. OBJECTIVES: The purpose of this work was to assess the association of household and child food insecurity with childhood overweight status. METHODS: The National Health and Nutrition Examination Survey 1999-2002 uses a stratified multistaged probability sample and collects a broad array of data from a nationally representative sample of US citizens. All children 3 to 17 years old in this sample are included in these analyses. We measured BMI categorized as at risk for overweight or greater (> or = 85%) or overweight (> or = 95%) and household and child food security/insecurity using the US Food Security Scale. RESULTS: When compared with children from food-secure households, children from food-insecure households were more likely to demonstrate significant associations with being at risk for overweight or greater in the following demographic categories: 12 to 17 years, girls, white, and in households with income < 100% and > 4 times the federal poverty level. Household food insecurity is associated with child overweight status in children aged 12 to 17, girls, and children who live in households with incomes > 4 times the federal poverty level. Child food insecurity demonstrated the same associations with being at risk for overweight or greater, as did household food insecurity, but associations were also seen in 3- to 5-year-old children, boys, and Mexican American children. Child food insecurity is significantly associated with child overweight status for children aged 12 to 17, girls, white children, and children in families with income < or = 100% poverty level. Controlling for ethnicity, gender, age, and family poverty index level, childhood food insecurity is associated with a child being at risk for overweight status or greater, but not overweight status. CONCLUSIONS: Household and child food insecurity are associated with being at risk for overweight and overweight status among many demographic categories of children. Child food insecurity is independently associated with being at risk for overweight status or greater while controlling for important demographic variables. Future longitudinal research is required to determine whether food insecurity is causally related to child overweight status. PMID- 17079543 TI - Racial/ethnic differences in breastfeeding initiation and continuation in the United kingdom and comparison with findings in the United States. AB - OBJECTIVE: Patterns of breastfeeding vary considerably across different racial/ethnic groups; however, little is known about factors that might explain differences across and within different racial/ethnic groups. Here we examine patterns of breastfeeding initiation and continuation among a racially/ethnically diverse sample of new mothers and compare this with patterns seen in the United States. The effects of demographic, social, economic, and cultural factors on racial/ethnic differences in breastfeeding practices are assessed. METHODS: The sample includes all singleton infants whose mothers participated in the first survey of the United Kingdom Millennium Cohort Study. Missing data reduced the sample to 17,474 (96%) infants with complete data. RESULTS: After adjustment for demographic, economic, and psychosocial factors, logistic regression models showed that Indian, Pakistani, Bangladeshi, black Caribbean, and black African mothers were more likely to initiate breastfeeding compared with white mothers. Further adjustment for a marker of cultural tradition attenuated these relationships, but all remained statistically significant, suggesting that some of the difference was a consequence of cultural factors. After adjustment for demographic, economic, and psychosocial factors, Indian, Pakistani, Bangladeshi, black Caribbean, and black African mothers were more likely to continue breastfeeding at 3 months compared with white mothers. Additional adjustment for a marker of cultural tradition attenuated the relationship for Indian, Pakistani, Bangladeshi, and black African mothers, but all remained statistically significant. Models run for breastfeeding continuation at 4 and 6 months were consistent with these results. CONCLUSIONS: We have shown that in the United Kingdom the highest breastfeeding rates are among black and Asian mothers, which is in stark contrast to patterns in the United States, where the lowest rate is seen among non-Hispanic black mothers. The contrasting racial/ethnic patterns of breastfeeding in the United Kingdom and United States necessitate very different public health approaches to reach national targets on breastfeeding and reduce health disparities. Those who implement future policies aimed at increasing breastfeeding rates need to pay attention to different social, economic, and cultural profiles of all racial/ethnic groups. PMID- 17079544 TI - Breastfeeding, sensitivity, and attachment. AB - OBJECTIVE: Our goal was to test the hypothesis that breastfeeding is associated with enhanced infant-mother attachment and its antecedent maternal sensitivity. METHODS: Breastfeeding intent and practice were assessed by questionnaires administered to 152 mothers between 32 weeks of gestation and 12 months postpartum. Early maternal sensitivity was measured by the Sensitivity to Cues subscale of the Nursing Child Assessment Satellite Training Feeding Scale at 3 months, and quality of the mother-infant interaction was measured by the Nursing Child Assessment Satellite Training Feeding Scale at 6 months. Security of attachment was evaluated by the Ainsworth Strange Situation at 12 months. RESULTS: A direct relationship between attachment security and breastfeeding practice was not identified. The quality of the mother-infant interaction at 6 months, rather than the type of feeding, predicted security of attachment. However, mothers who chose to breastfeed displayed greater sensitivity in dyadic interactions with their infants 3 months postnatally than those who chose to bottle feed, and intended breastfeeding duration prenatally correlated with sensitivity 3 months postpartum. Although a path analysis failed to demonstrate contributions of early breastfeeding duration to either sensitivity or security, it did substantiate a significant path between prenatal breastfeeding intent and attachment security mediated by sensitivity. In addition, multivariate analyses revealed that early sensitivity among breastfeeding mothers was an independent predictor of the duration of any and exclusive breastfeeding during the first year. CONCLUSIONS: Although the quality of the dyadic interaction in infancy, rather than feeding type, is predictive of attachment security, mothers who choose to breastfeed display enhanced sensitivity during early infancy that, in turn, may foster secure attachment. Among breastfeeding mothers, higher sensitivity is associated with longer duration of breastfeeding during the first postpartum year. These findings suggest a link between attachment security and breastfeeding. PMID- 17079545 TI - Breastfeeding and verbal ability of 3-year-olds in a multicity sample. AB - OBJECTIVES: Our goal was to analyze the effect of maternal verbal ability and education on the association between breastfeeding and children's cognitive functioning. First, we hypothesized that maternal verbal abilities account for a large portion of the association between breastfeeding and child verbal abilities. Second, we hypothesized that after adjusting for maternal verbal abilities, a positive effect of breastfeeding will be most evident among highly educated mothers, because these mothers may have more opportunity to engage in cognitively stimulating parenting than do mothers with less education. PATIENTS AND METHODS: With data on 1645 American-born mothers participating in a longitudinal birth cohort study, we used linear regression to determine the influence of breastfeeding for at least 1 month on the Peabody Picture Vocabulary Test-Third Edition scores of 3-year-old children. Models were adjusted for an extensive set of demographic characteristics, including mother's Peabody Picture Vocabulary Test and the Home Observation for Measurement of the Environment score. Mothers were categorized into 1 of 3 educational-status groups: no high school diploma, high school diploma, and some post-secondary education. RESULTS: In unadjusted mean comparisons, breastfed children had Peabody Picture Vocabulary Test scores that were 6.6 points higher than children who were not breastfed. After adjusting for demographic characteristics and maternal verbal ability, the coefficient dropped to 1.72. Among mothers with education beyond high school, the children's Peabody Picture Vocabulary Test scores in adjusted models were 2.2 points higher for breastfed children. Among mothers with a high school diploma or less, there were no significant differences in the children's Peabody Picture Vocabulary Test scores by breastfeeding status. These results were consistent in white, black, and Hispanic children. CONCLUSIONS: Maternal Peabody Picture Vocabulary Test scores mediate much of the association between breastfeeding and child verbal abilities. The beneficial effects of breastfeeding on children's cognition may emerge only when breastfeeding is done in conjunction with other positive parenting behaviors. The advantageous effects of breastfeeding do not seem to be solely attributable to the superior nutrient content of breast milk. PMID- 17079546 TI - Growth in 10- to 12-year-old children born at 23 to 25 weeks' gestation in the 1990s: a Swedish national prospective follow-up study. AB - BACKGROUND: Knowledge of long-term growth of extremely preterm infants in relation to gestational age is incomplete, and there are concerns regarding their poor growth in early childhood. As part of a longitudinal study of a national cohort of infants born at <26 weeks' gestation (extremely immature), growth development from birth to the age of 11 years was examined, and correlates of growth attainment were analyzed. METHODS: Two hundred forty-seven extremely immature children were born alive from April 1990 through March 1992 in the whole of Sweden, and 89 (36%) survived. Growth and neurosensory outcomes of all extremely immature survivors were evaluated at 36 months of age. Eighty-six (97%) extremely immature children were identified and assessed at 11 years of age. In this growth study, 83 extremely immature infants (mean [SD]: birth weight, 772 g [110 g]; gestational age, 24.6 weeks [0.6 weeks]) without severe motor disability were followed up prospectively from birth to 11 years old and compared with a matched group of 83 children born at term. z scores for weight, height, head circumference, and BMI were computed for all children. We also examined gender specific longitudinal growth measures. Predictors of 11-year growth were studied by multivariate analyses. RESULTS: Extremely immature children were significantly smaller in all 3 growth parameters than the controls at 11 years. Extremely immature children showed a sharp decline in weight and height z scores up to 3 months' corrected age, followed by catch-up growth in both weight and height up to 11 years. In contrast to weight and height, extremely immature children did not exhibit catch-up growth in head circumference after the first 6 months of life. The mean BMI z scores increased significantly from 1 to 11 years in both groups. The mean BMI change between 1 and 11 years of age was significantly larger in extremely immature than in control participants. Extremely immature girls showed a faster weight increase than extremely immature boys, whereas catch up growth in height and head circumference was similar in these groups. Multiple regression analyses revealed that preterm birth and parental height were significant predictors of 11-year height, and group status (prematurity) correlated strongly with head circumference. CONCLUSIONS: Children born at the limit of viability attain poor growth in early childhood, followed by catch-up growth to age 11 years, but remain smaller than their term-born peers. Strategies that improve early growth might improve the outcome. PMID- 17079547 TI - Chronic conditions, functional limitations, and special health care needs in 10- to 12-year-old children born at 23 to 25 weeks' gestation in the 1990s: a Swedish national prospective follow-up study. AB - BACKGROUND: Children born extremely immature (gestational age < 26 weeks' gestation) increasingly reach school age. Information on their overall functioning and special health care needs is necessary to plan for their medical and educational services. This study was undertaken to examine neurosensory, medical, and developmental conditions together with functional limitations and special health care needs of extremely immature children compared with control subjects born at term. METHODS: We studied 11-year-old children born before 26 completed weeks of gestation in all of Sweden from 1990 through 1992. All had been evaluated at 36 months' corrected age. Identification of children with chronic conditions lasting > or = 12 months was based on a questionnaire administered to parents. Neurosensory impairments were identified by reviewing health records. Information regarding other specific medical diagnoses and developmental disabilities was obtained by standard parent and teacher questionnaires. RESULTS: Of 89 eligible children, 86 (97%) were studied at a mean age of 11 years. An equal number of children born at term served as controls. Logistic-regression analyses adjusting for social risk factors and gender showed that significantly more extremely immature children than controls had chronic conditions, including functional limitations (64% vs 11%, respectively), compensatory dependency needs (59% vs 25%), and services above those routinely required by children (67% vs 22%). Specific diagnoses or disabilities with higher rates in extremely immature children than in controls included neurosensory impairment (15% vs 2%), asthma (20% vs 6%), poor motor skills of > 2 SDs above the mean (26% vs 3%), poor visual perception of > 2 SDs above the mean (21% vs 4%), poor learning skills of > 2 SDs above the mean (27% vs 3%), poor adaptive functioning with T scores of < 40 (42% vs 9%), and poor academic performance with T score < 40 (49% vs 7%). CONCLUSIONS: Children born extremely immature have significantly greater health problems and special health care needs at 11 years of age. However, few children have severe impairments that curtail major activities of daily living. PMID- 17079548 TI - The clinical spectrum of developmental language impairment in school-aged children: language, cognitive, and motor findings. AB - OBJECTIVE: Our goal was to evaluate detailed school-age language, nonverbal cognitive, and motor development in children with developmental language impairment compared with age-matched controls. METHODS: Children with developmental language impairment or normal language development (controls) aged 7 to 13 years were recruited. Children underwent language assessment (Clinical Evaluation of Language Fundamentals-4, Peabody Picture Vocabulary-3, Goldman Fristoe Test of Articulation-2), nonverbal cognitive assessment (Wechsler Intelligence Scale for Children-IV), and motor assessment (Movement Assessment Battery for Children). Exclusion criteria were nonverbal IQ below the 5th percentile or an acquired language, hearing, autistic spectrum, or neurologic disorder. RESULTS: Eleven children with developmental language impairment (7:4 boys/girls; mean age: 10.1 +/- 0.8 years) and 12 controls (5:7 boys/girls; mean age: 9.5 +/- 1.8 years) were recruited. Children with developmental language impairment showed lower mean scores on language (Clinical Evaluation of Language Fundamentals-4--developmental language impairment: 79.7 +/- 16.5; controls: 109.2 +/- 9.6; Goldman-Fristoe Test of Articulation-2--developmental language impairment: 94.1 +/- 10.6; controls: 104.0 +/- 2.8; Peabody Picture Vocabulary-3- developmental language impairment: 90.5 +/- 13.8; controls: 100.1 +/- 11.6), cognitive (Wechsler Intelligence Scale for Children-IV--developmental language impairment: 99.5 +/- 15.5; controls: 113.5 +/- 11.9), and motor measures (Movement Assessment Battery for Children percentile--developmental language impairment: 12.7 +/- 16.7; controls: 66.1 +/- 30.6) and greater discrepancies between cognitive and language scores (Wechsler Intelligence Scale for Children IV/Clinical Evaluation of Language Fundamentals-4--developmental language impairment: 17.8 +/- 17.8; controls: 1.2 +/- 12.7). Motor impairment was more common in children with developmental language impairment (70%) than controls (8%). CONCLUSIONS: Developmental language impairment is characterized by a broad spectrum of developmental impairments. Children identified on the basis of language impairment show significant motor comorbidity. Motor assessment should form part of the evaluation and follow-up of children with developmental language impairment. PMID- 17079549 TI - Hyperglycemia is a risk factor for early death and morbidity in extremely low birth-weight infants. AB - OBJECTIVES: The objectives of this study were to determine the prevalence of hyperglycemia in extremely low birth-weight infants and to determine whether hyperglycemia increases the risk of early adverse outcomes (death or intraventricular hemorrhage of grade 3 or 4) and/or affects the length of hospital stay among survivors without intraventricular hemorrhage. METHODS: The charts of all extremely low birth-weight infants (n = 93) admitted to Texas Children's Hospital (Houston, TX) during 2001 were reviewed. The highest daily blood glucose concentrations, highest dopamine infusion rates, highest daily percentage of inspired oxygen, and mean blood sodium concentrations were averaged over the first week of life or before death or occurrence of grade 3 or 4 intraventricular hemorrhage. Among survivors without severe intraventricular hemorrhage, the time ratio for blood glucose concentrations of >150 mg/dL was calculated. RESULTS: More than 50% of the infants had persistent blood glucose concentrations of >150 mg/dL during their first week of life. Early adverse outcomes were associated with the average highest daily blood glucose concentration through interaction with the Clinical Risk Index for Babies score and with the average highest daily percentage of inspired oxygen. The length of hospital stay was associated with the time ratio for blood glucose concentrations of >150 mg/dL through interaction with birth weight and the average highest daily percentage of inspired oxygen. CONCLUSION: These data confirm the high prevalence of hyperglycemia among parenterally fed, extremely low birth-weight infants and show that high blood glucose concentrations increase the risk of early death and grade 3 or 4 intraventricular hemorrhage and the length of hospital stay among survivors without intraventricular hemorrhage, which suggests that prevention and treatment of hyperglycemia may improve the outcomes of extremely low birth-weight infants. PMID- 17079550 TI - Health of children born as a result of in vitro fertilization. AB - OBJECTIVE: The purpose of this study was to use nationwide registries to examine the health of children up to 4 years of age who were born as a result of in vitro fertilization. METHODS: Children born after in vitro fertilization (N = 4559) from 1996 to 1999 were monitored until 2003. Two control groups were selected from the Finnish Medical Birth Register as follows: all other children (excluding children born after ovulation induction) from the same period (N = 190,398, for study of perinatal health and hospitalizations) and a random sample of those children (n = 26,877, for study of health-related benefits). Mortality rates and odds ratios for perinatal outcomes, hospitalizations, health-related benefits, and long-term medication use were calculated. RESULTS: Although the health of most in vitro fertilization children was good, such children had more health problems than other children. A total of 35.7% of in vitro fertilization children and 2.2% of control children were multiple births, and the health of multiple births was worse than that of singletons. Perinatal outcomes of in vitro fertilization children were worse and hospital episodes were more common than among control children. Risks for cerebral palsy and psychological and developmental disorders were increased. Among in vitro fertilization singletons, worse results for perinatal outcomes and hospitalizations, but no increased risk for specific diseases, were found. The health of in vitro fertilization multiple births was comparable to the health of control multiple births. CONCLUSIONS: Reducing the number of transferred embryos would improve the health of in vitro fertilization children. Additional studies are needed to explain the poorer health of in vitro fertilization singletons, as well as follow-up studies to examine the health of in vitro fertilization children from 4 years onward. PMID- 17079551 TI - National trends in the use of antireflux procedures for children. AB - OBJECTIVE: The purpose of this study was to analyze recent nationwide trends in the use of and outcomes after antireflux surgery for children. METHODS: We conducted a retrospective cohort study of children (age: <18 years) undergoing antireflux surgery by using data from 1996 to 2003 from the Nationwide Inpatient Sample. Census data were used to calculate the population-based rates of procedures stratified according to age and presence of neurologic impairment. Multivariate analyses were performed to determine factors associated with length of stay and in-hospital death. RESULTS: During the study period, 48,665 antireflux procedures were performed for children in the US. Although procedure rates were generally higher in 2003 than in 1996, no trends in rates were observed among different age groups and census regions during the study period. The highest population-based procedure rates were observed among infants (49-101 procedures per 100,000 population). There was a significant decrease in the percentages of children undergoing antireflux procedures who were neurologically impaired between 1996 and 2003 (53% vs 40%). Neurologically impaired children had longer lengths of stay and higher mortality rates than did neurologically normal children. CONCLUSIONS: Although procedure rates have not changed, the use of antireflux surgery has evolved during the laparoscopic era, with a decreasing percentage of neurologically impaired children undergoing this procedure. Antireflux procedures were performed predominantly for infants, most of whom were neurologically normal. Neurologically impaired children remain a group at high risk for death after antireflux procedures. PMID- 17079552 TI - Primary care physicians' attitudes regarding follow-up care for children with positive newborn screening results. AB - BACKGROUND: Although primary care physicians are responsible for providing follow up care after a positive newborn screen, little is known about their willingness or ability to do so. METHODS: A national mail survey of a random sample of 350 general pediatricians and 350 family physicians was conducted from April to June 2006. RESULTS: The response rate was 63% among pediatricians and 50% among family physicians. Most pediatricians (89.7%) and nearly one half of family physicians (44.1%) had had a patient with a positive newborn screen within the past 5 years. Most respondents thought that primary care physicians should be responsible for informing families about a positive newborn screen (73.2%), arranging confirmatory testing (66.0%), and coordinating subspecialty referral (85.3%). However, more than one half (56.2%) would prefer newborn screening programs to provide the initial evaluation of positive newborn screening results. Some respondents (but fewer pediatricians than family physicians) reported that they were not competent to discuss conditions included in newborn screening panels (eg, 22.6% of pediatricians and 53.2% of family physicians for phenylketonuria and 8.8% of pediatricians and 40.4% of family physicians for congenital hypothyroidism). More than one half (58.3%) thought that families with a child diagnosed as having congenital hypothyroidism should receive formal genetic counseling. Respondents were less likely to think that families with a child with sickle cell trait, compared with families with a child who is a cystic fibrosis carrier, should receive formal genetic counseling (69.3% vs 84.1%). CONCLUSIONS: Many primary care physicians are not prepared to manage the follow-up care of children with a positive newborn screen, including initial counseling, diagnosis, and subspecialty referral. New strategies are needed to ensure appropriate and equitable health care delivery. PMID- 17079553 TI - Slight/mild sensorineural hearing loss in children. AB - OBJECTIVE: The goal was to determine the prevalence and effects of slight/mild bilateral sensorineural hearing loss among children in elementary school. METHODS: A cross-sectional, cluster-sample survey of 6581 children (response: 85%; grade 1: n = 3367; grade 5: n = 3214) in 89 schools in Melbourne, Australia, was performed. Slight/mild bilateral sensorineural hearing loss was defined as a low-frequency pure-tone average across 0.5, 1, and 2 kHz and/or a high-frequency pure-tone average across 3, 4, and 6 kHz of 16 to 40 dB hearing level in the better ear, with air/bone-conduction gaps of < 10 dB. Parents reported children's health-related quality of life and behavior. Each child with slight/mild bilateral sensorineural hearing loss, matched to 2 normally hearing children (low frequency pure-tone average and high-frequency pure-tone average of < or = 15 dB hearing level in both ears), completed standardized assessments. Whole-sample comparisons were adjusted for type of school, grade level, and gender, and matched-sample comparisons were adjusted for nonverbal IQ scores. RESULTS: Fifty five children (0.88%) had slight/mild bilateral sensorineural hearing loss. Children with and without sensorineural hearing loss scored similarly in language (mean: 97.2 vs 99.7), reading (101.1 vs 102.8), behavior (8.4 vs 7.0), and parent and child-reported child health-related quality of life (77.6 vs 80.0 and 76.1 vs 77.0, respectively), but phonologic short-term memory was poorer (91.0 vs 102.8) in the sensorineural hearing loss group. CONCLUSIONS: The prevalence of slight/mild bilateral sensorineural hearing loss was lower than reported in previous studies. There was no strong evidence that slight/mild bilateral sensorineural hearing loss affects adversely language, reading, behavior, or health-related quality of life in children who are otherwise healthy and of normal intelligence. PMID- 17079554 TI - Should our well-child care system be redesigned? A national survey of pediatricians. AB - OBJECTIVE: The goal was to examine pediatricians' views about whether and how well-child care for children 0 to 5 years of age should be changed. METHODS: A mail survey of a national random sample of 1000 general pediatricians was performed with a survey instrument that examined pediatricians' attitudes and behaviors toward our current way and an ideal way of providing well-child care. Results were analyzed for the following 3 major domains of change in well-child care: provider type, visit format, and visit location. RESULTS: Sixty percent (n = 502) of eligible subjects responded to the survey. Nearly all respondents (97%) rated the current US system as excellent or good in providing well-child care. Most pediatricians (85%-91%) reported that they are currently the main providers of anticipatory guidance, developmental screening, and psychosocial screening. However, a majority (54%-60%) reported that, in an ideal system that maximized the effectiveness and efficiency of care, nonphysicians would provide these services. Fewer pediatricians (24%) reported that ideally nonphysicians should provide the physical examination. The majority of respondents (79%-93%) reported that at least some anticipatory guidance, minor acute care, and chronic care services could be conducted through telephone or e-mail communication, and 55% stated that at least some well-child care services should be provided in alternative locations, such as day care centers. In multivariate analysis, support for these changes was distributed widely across pediatricians with varying personal and practice characteristics. CONCLUSIONS: Although most pediatricians are generally satisfied with our current way of providing well child care, a majority think that a system that is less reliant on physicians and face-to-face office visits would be a more effective and efficient way to provide care. PMID- 17079555 TI - Genetic susceptibility to retinopathy of prematurity. AB - OBJECTIVES: The goals were to isolate and to estimate the genetic susceptibility to retinopathy of prematurity. METHODS: A retrospective study (1994-2004) from 3 centers was performed with zygosity data for premature twins who were born at a gestational age of < or = 32 weeks and survived beyond a postmenstrual age of 36 weeks. Retinopathy of prematurity was diagnosed and staged by pediatric ophthalmologists at each center. Data analyses were performed with mixed-effects logistic regression analysis and latent variable probit modeling. RESULTS: A total of 63 monozygotic and 137 dizygotic twin pairs were identified and analyzed. Data on gestational age, birth weight, gender, respiratory distress syndrome, retinopathy of prematurity, bronchopulmonary dysplasia, duration of ventilation and supplemental oxygen use, and length of stay were comparable between monozygotic and dizygotic twins. In the mixed-effects logistic regression analysis for retinopathy of prematurity, gestational age and duration of supplemental oxygen use were significant covariates. After controlling for known and unknown nongenetic factors, genetic factors accounted for 70.1% of the variance in liability for retinopathy of prematurity. CONCLUSION: In addition to prematurity and environmental factors, there is a strong genetic predisposition to retinopathy of prematurity. PMID- 17079556 TI - Impact of umbilical cord cleansing with 4.0% chlorhexidine on time to cord separation among newborns in southern Nepal: a cluster-randomized, community based trial. AB - OBJECTIVE: Within a community-based, cluster-randomized study of the effects of 4.0% chlorhexidine on omphalitis and mortality risk, we aimed to describe the distribution of times to separation and the impact of topical chlorhexidine treatment on cord-separation times. METHODS: Between November 2002 and March 2005, 15123 infants were assigned randomly within communities in southern Nepal to receive 1 of the following 3 cord-care regimens: cleansing with 4.0% chlorhexidine, cleansing with soap and water, or dry cord care. In intervention clusters, field workers cleansed the cord in the home on days 1, 2, 3, 4, 6, 8, and 10 after birth. Newborns were monitored throughout the newborn period for signs of omphalitis, and the time to cord separation was noted. Separation times were compared across treatment groups. Cord infection risk and a range of infant and household characteristics were assessed for their relationships to separation time. RESULTS: The mean separation time was shorter in dry cord care (4.24 days) and soap/water (4.25 days) clusters than in chlorhexidine clusters (5.32 days; mean difference: 1.08 days). Cords of infants who received chlorhexidine were 3.6 times more likely to separate after 7 days. Separation time was not associated with omphalitis. Home-delivered topical antiseptics, facility-based birth, and birth attendant hand-washing were associated with greater likelihoods of cord separation after 7 days of age. CONCLUSIONS: In this setting, the umbilical cord separated more rapidly than observed in hospital-based studies, and the impact of chlorhexidine cleansing on separation times was negligible. Increased cord separation time attributable to topical chlorhexidine treatment should not be considered a factor in decision-making in settings where the baseline risk of omphalitis is high and chlorhexidine might reduce infection and mortality risks significantly. PMID- 17079557 TI - Medication errors related to computerized order entry for children. AB - OBJECTIVE: The objective of this study was to determine the frequency and types of pediatric medication errors attributable to design features of a computerized order entry system. METHODS: A total of 352 randomly selected, inpatient, pediatric admissions were reviewed retrospectively for identification of medication errors, 3 to 12 months after implementation of computerized order entry. Errors were identified and classified by using an established, comprehensive, active surveillance method. Errors attributable to the computer system were classified according to type. RESULTS: Among 6916 medication orders in 1930 patient-days, there were 104 pediatric medication errors, of which 71 were serious (37 serious medication errors per 1000 patient-days). Of all pediatric medication errors detected, 19% (7 serious and 13 with little potential for harm) were computer related. The rate of computer-related pediatric errors was 10 errors per 1000 patient-days, and the rate of serious computer-related pediatric errors was 3.6 errors per 1000 patient-days. The following 4 types of computer-related errors were identified: duplicate medication orders (same medication ordered twice in different concentrations of syrup, to work around computer constraints; 2 errors), drop-down menu selection errors (wrong selection from a drop-down box; 9 errors), keypad entry error (5 typed instead of 50; 1 error), and order set errors (orders selected from a pediatric order set that were not appropriate for the patient; 8 errors). In addition, 4 preventable adverse drug events in drug ordering occurred that were not considered computer related but were not prevented by the computerized physician order entry system. CONCLUSIONS: Serious pediatric computer-related errors are uncommon (3.6 errors per 1000 patient-days), but computer systems can introduce some new pediatric medication errors that are not typically seen in a paper ordering system. PMID- 17079558 TI - Asthma guideline use by pediatricians in private practices and asthma morbidity. AB - OBJECTIVE: We sought to examine whether pediatric clinicians in private, non health maintenance organization practices could implement the national asthma guidelines and whether, when implemented, these guidelines would decrease medical services utilization and improve asthma care for children. METHODS: A trial of a disease management program (Easy Breathing II) involving 20 private pediatric practices in the greater Hartford, Connecticut area was conducted between January 1, 2001, and December 31, 2003. Demographic data on participating practitioners and patients were obtained from questionnaires. Medical services utilization data from claims were obtained from ConnectiCare, a regional managed care organization. RESULTS: Of the 16750 children enrolled in Easy Breathing II, 2458 were enrolled in ConnectiCare and 490 had asthma. Inhaled corticosteroid use increased in the community overall during the study period. After enrollment in Easy Breathing II, with adjustment for age, gender, ethnicity, asthma severity, season, and calendar year, children with persistent asthma experienced an additional 47% increase in inhaled corticosteroid use, a 56% reduction in outpatient visits, and a 91% decrease in emergency department visits for treatment of asthma. Adherence to national asthma guidelines for prescribing inhaled corticosteroids was 95%. Seventeen of the 20 practices are still using Easy Breathing, 5 years after program implementation. CONCLUSIONS: Pediatric primary care clinicians in private practice settings can implement an asthma management program patterned after the national asthma guidelines. When implemented, this program is successful in reducing medical services utilization for children with asthma. Just as differences in patterns of medical services utilization exist in private practices, compared with urban clinics, the impact of disease management on medical services utilization differs in private practices, compared with urban clinics. PMID- 17079559 TI - Diabetes mellitus screening in pediatric primary care. AB - OBJECTIVE: The goal was to determine the rates of diabetes screening and the prevalence of screening abnormalities in overweight and nonoverweight individuals in an urban primary care clinic. METHODS: This study was a retrospective chart review conducted in a hospital-based urban primary care setting. Deidentified data for patients who were 10 to 19 years of age and had > or = 1 BMI measurement between September 1, 2002, and September 1, 2004, were extracted from the hospital electronic health record. RESULTS: A total of 7710 patients met the study criteria. Patients were 73.0% black or Hispanic and 47.0% female; 42.0% of children exceeded normal weight, with 18.2% at risk for overweight and 23.8% overweight. On the basis of BMI, family history, and race, 8.7% of patients met American Diabetes Association criteria for type 2 diabetes mellitus screening, and 2452 screening tests were performed for 1642 patients. Female gender, older age group, and family history of diabetes were associated with screening. Increasing BMI percentile was associated with screening, exhibiting a dose response relationship. Screening rates were significantly higher (45.4% vs 19.0%) for patients who met the American Diabetes Association criteria; however, less than one half of adolescents who should have been screened were screened. Abnormal glucose metabolism was seen for 9.2% of patients screened. CONCLUSIONS: This study shows that, although pediatricians are screening for diabetes mellitus, screening is not being conducted according to the American Diabetes Association consensus statement. Point-of-care delivery of consensus recommendations could increase provider awareness of current recommendations, possibly improving rates of systematic screening and subsequent identification of children with laboratory evidence of abnormal glucose metabolism. PMID- 17079560 TI - Mental health screening in pediatric practice: factors related to positive screens and the contribution of parental/personal concern. AB - OBJECTIVES: The goals were to examine factors related to positive Pediatric Symptom Checklist scores in an urban practice and to examine the relative contribution of parental/personal concern about emotional and behavioral problems to mental health problem identification. METHODS: Annual screening using the Pediatric Symptom Checklist was implemented in Cambridge Pediatrics (Cambridge, MA). A social worker was colocated in the clinic to provide therapeutic interventions for patients. A sample of 1668 screened patients between 4 years 11 months and 19 years of age was used for analysis. Bivariate and multivariate analyses were conducted to determine factors predictive of positive Pediatric Symptom Checklist scores, including demographics, socioeconomic indicators, enrollment in counseling, and parental/personal concern. Parental/personal concern, counseling, and positive Pediatric Symptom Checklist scores were examined to determine their efficacy as screening methods. RESULTS: Six percent of the population had positive Pediatric Symptom Checklist scores. There were statistically significant relationships between a positive score and being in counseling, parental/personal concern, having public insurance, and living in an area with median household incomes of less than 50,000 dollars. Parental/personal concern was 40% sensitive for a positive score. A positive Pediatric Symptom Checklist score with or without parental/personal concern identified 3.8% of the population; parental/personal concern with or without a positive Pediatric Symptom Checklist score identified 4.5%. CONCLUSIONS: Mental health screening can be effectively implemented in a pediatric practice. Colocated mental health professionals provide additional support. The combination of a screening tool and questions about parental/personal concern and present counseling can provide critical information about a child's mental health. PMID- 17079562 TI - Primary hemorrhagic stroke in children with sickle cell disease is associated with recent transfusion and use of corticosteroids. AB - OBJECTIVES: Primary hemorrhagic stroke is an uncommon complication of sickle cell disease, with reported mortality rates of 24% to 65%. Most reported cases are in adults; little is known about its occurrence in children. Proposed risk factors include previous ischemic stroke, aneurysms, low steady-state hemoglobin, high steady-state leukocyte count, acute chest syndrome, and hypertransfusion. We performed a retrospective case-control study to evaluate risk and prognostic factors for primary hemorrhagic stroke among children with sickle cell disease. PATIENTS AND METHODS: Case subjects (sickle cell disease and primary hemorrhagic stroke) and control subjects (sickle cell disease and ischemic stroke) were identified at 2 children's hospitals from January 1979 to December 2004 by reviewing divisional records and the discharge databases. RESULTS: We identified 15 case subjects (mean age: 10.4 +/- 1.3 years) and 29 control subjects (mean age: 5.2 +/- 0.4 years). An increased risk of hemorrhagic stroke was associated with a history of hypertension and recent (in the last 14 days) transfusion, treatment with corticosteroids, and possibly nonsteroidal antiinflammatory drugs. Average blood pressures at well visits (adjusted for age and gender) were similar between the 2 groups, suggesting that hypertension was intermittent. CONCLUSIONS: In this group of children with sickle cell disease, hemorrhagic stroke was associated with a history of hypertension or antecedent events including transfusion or treatment with corticosteroids. Improved understanding of risk and prognostic factors, especially those that are modifiable, may help prevent this devastating complication in children with sickle cell disease. PMID- 17079561 TI - Childhood depressive symptoms and early onset of alcohol use. AB - OBJECTIVES: Few studies have assessed the relationship between depressive symptoms and early onset of alcohol use in children and early adolescents. We aimed to determine whether depressive symptoms in children are associated with subsequent initiation of alcohol use and, if so, whether this association is merely a result of demographic, parental, and/or individual risk factors shared by depression and alcohol use or independent of these shared risk factors. METHODS: Analyses were based on a subsample of 10- to 13-year-old children (N = 1119) from the Boricua Youth Study, a longitudinal study of psychopathology among Puerto Rican children and early adolescents. Children in the study were assessed over 3 waves between 2000 and 2004. In-person structured interviews were conducted with both parents and children. RESULTS: Depressive symptoms and alcohol use shared some significant risk and protective factors, such as parental psychopathology, parenting, child exposure to violence, and antisocial behaviors. After controlling for these factors, the association between depressive symptoms and alcohol use was reduced, but childhood depressive symptoms were still positively associated with subsequent alcohol use initiation. Children with medium or high levels of depressive symptoms were more than twice as likely to use alcohol as those with <2 depressive symptoms. CONCLUSIONS: The finding of the current study that early life depressive symptoms may lead to earlier onset of alcohol use indicates the importance of identifying and treating depressive symptoms in preadolescent children. It also demonstrates the importance of examining shared risk and protective factors for understanding the relationship between depressive symptoms and alcohol use. PMID- 17079563 TI - Recurrent respiratory papillomatosis in children: masquerader of common respiratory diseases. AB - BACKGROUND: Recurrent respiratory papillomatosis in children is an uncommon but potentially life-threatening benign tumor of the respiratory tract with laryngeal predilection. The diagnosis of recurrent respiratory papillomatosis may be challenging unless there is a high index of suspicion and awareness of the variable presentations. METHODS: We reviewed the medical charts of children with recurrent respiratory papillomatosis treated at a tertiary children's hospital. The presentation of recurrent respiratory papillomatosis is illustrated by a series of case reports. We provide a paradigm to assist in the early diagnosis of children with recurrent respiratory papillomatosis. RESULTS: Five patients, aged 2 to 6 years, were erroneously diagnosed with recurrent croup, asthma, laryngeal hemangioma, and tracheomalacia after presenting with variable degrees of chronic dyspnea, cough, stridor, dysphonia, weak cry, and syncope. Once the diagnosis of recurrent respiratory papillomatosis was made, recurring surgical ablation of papillomata was initiated. CONCLUSIONS: Any child presenting with a voice disturbance with or without stridor is recommended to have diagnostic flexible fiber-optic laryngoscopy. Recurrent respiratory papillomatosis should be considered in children when other common pediatric airway diseases either do not follow the natural history or do not respond to treatment of the common disorder. PMID- 17079564 TI - Parental rules and monitoring of children's movie viewing associated with children's risk for smoking and drinking. AB - OBJECTIVES: Evidence suggests that media portrayals of tobacco and alcohol use are important predictors of adolescent smoking and drinking. We examined the role of parents in monitoring and limiting children's movie exposure and whether or not this was associated with a lower risk of adolescent smoking and drinking. DESIGN: We surveyed 2606 child-parent dyads between 2002 and 2003. We asked children (9-12 years of age) how often their parents engaged in specific behaviors to monitor their movie viewing and if their parents allowed them to watch R-rated movies. We also surveyed children about parental monitoring of nonmedia-related behaviors. The primary outcomes were risk of smoking and drinking alcohol, defined by attitudinal susceptibility or early experimentation with either substance. RESULTS: Less than half (45.0%) the children were prohibited from watching R-rated movies. Of those who were allowed to watch R rated movies, one third (34.7%) always viewed them with a parent and two thirds (65.3%) sometimes watched them without a parent. Less than 10% of the children reported that their parents consistently engaged in all 4 movie-monitoring behaviors. Even after controlling for parental monitoring of nonmedia-related behaviors and other covariates, children were at lower risk of smoking and drinking if their parents prohibited them from watching R-rated movies. Parental coviewing of R-rated movies was associated with a lower risk of child smoking but not drinking if parents consistently monitored what their children watched. CONCLUSIONS: Parental rules and monitoring of children's movie viewing may have a protective influence on children's risk for smoking and drinking, over and above parental monitoring of nonmedia related behaviors. This highlights a potential role for parents in preventing early initiation of tobacco and alcohol use among adolescents. PMID- 17079565 TI - The impact of early nutrition in premature infants on later childhood insulin sensitivity and growth. AB - OBJECTIVES: Children born prematurely have decreased insulin sensitivity. The etiology of this insulin resistance is unknown. The aim of this study was to evaluate infant nutrition and its influence on insulin sensitivity and postnatal growth in children born < or = 32 weeks' gestation. METHODS: A total of 56 healthy, developmentally normal, prepubertal children, aged 4 to 10 years were recruited. Thirty-seven were born < or = 32 weeks' gestation, and 19 were control subjects born at term with a birth weight > 10th percentile. Insulin sensitivity (10(-4) min(-1) microU/mL) was calculated from a 90-minute frequently sampled intravenous glucose tolerance test. Perinatal, nutritional, and growth data were obtained retrospectively from both neonatal and early infancy records in the premature cohort. RESULTS: Children born prematurely had decreased insulin sensitivity when compared with those born at term (13.8 vs 30.6). Neonatal nutrition was not correlated with insulin sensitivity; however, all of the infants had inadequate protein in the first month followed by excessive fat intake thereafter. Premature children with greater weight gain had lower insulin sensitivity. Higher carbohydrate intake in the first month of life was associated with greater weight gain from birth. No relationship was seen between weight gain and either protein or lipid intake. CONCLUSIONS: Prematurely born children are insulin resistant and have suboptimal neonatal nutrition. Greater childhood weight gain magnifies this reduction in insulin sensitivity and seems to be associated with early nutrition. We speculate that a high carbohydrate neonatal diet may lead to greater weight gain and a greater reduction in insulin sensitivity in this group. PMID- 17079566 TI - Vitamin D status in children and young adults with inflammatory bowel disease. AB - OBJECTIVES: Previous studies of vitamin D status in pediatric patients with inflammatory bowel disease have revealed conflicting results. We sought to report (1) the prevalence of vitamin D deficiency (serum 25-hydroxy-vitamin D concentration < or = 15 ng/mL) in a large population with inflammatory bowel disease, (2) factors predisposing to this problem, and (3) its relationship to bone health and serum parathyroid hormone concentration. PATIENTS AND METHODS: A total of 130 patients (8-22 years of age) with inflammatory bowel disease, 94 with Crohn disease and 36 with ulcerative colitis, had serum 25-hydroxy-vitamin D, intact parathyroid hormone, and lumbar spine bone mineral density (using dual energy x-ray absorptiometry) measured at Children's Hospital Boston. RESULTS: The prevalence of vitamin D deficiency was 34.6%. Mean serum 25-hydroxy-vitamin D concentration was similar in patients with Crohn disease and ulcerative colitis, 52.6% lower among patients with dark skin complexion, 33.4% lower during the winter months (December 22 to March 21), and 31.5% higher among patients who were taking vitamin D supplements. Serum 25-hydroxy-vitamin D concentration was positively correlated with weight and BMI z score, disease duration, and serum albumin concentration and negatively correlated with erythrocyte sedimentation rate. Patients with Crohn disease and upper gastrointestinal tract involvement were more likely to be vitamin D deficient than those without it. Serum 25 hydroxy-vitamin concentration was not associated with lumbar spine bone mineral density z score or serum parathyroid hormone concentration. CONCLUSIONS: Vitamin D deficiency is highly prevalent among pediatric patients with inflammatory bowel disease. Factors predisposing to the problem include having a dark-skin complexion, winter season, lack of vitamin D supplementation, early stage of disease, more severe disease, and upper gastrointestinal tract involvement in patients with Crohn disease. The long-term significance of hypovitaminosis D for this population is unknown at present and merits additional study. PMID- 17079567 TI - An effective nonchemical treatment for head lice: a lot of hot air. AB - OBJECTIVES: Head lice (Pediculus humanus capitis) are a major irritant to children and their parents around the world. Each year millions of children are infested with head lice, a condition known as pediculosis, which is responsible for tens of millions of lost school days. Head lice have evolved resistance to many of the currently used pediculicides; therefore, an effective new treatment for head lice is needed. In this study we examined the effectiveness of several methods that use hot air to kill head lice and their eggs. METHODS: We tested 6 different treatment methods on a total of 169 infested individuals. Each method delivers hot air to the scalp in a different way. We evaluated how well these methods kill lice and their eggs in situ. We also performed follow-up inspections to evaluate whether the sixth, most successful, method can cure head louse infestations. RESULTS: All 6 methods resulted in high egg mortality (> or = 88%), but they showed more-variable success in killing hatched lice. The most successful method, which used a custom-built machine called the LouseBuster, resulted in nearly 100% mortality of eggs and 80% mortality of hatched lice. The LouseBuster was effective in killing lice and their eggs when operated at a comfortable temperature, slightly cooler than a standard blow-dryer. Virtually all subjects were cured of head lice when examined 1 week after treatment with the LouseBuster. There were no adverse effects of treatment. CONCLUSIONS: Our findings demonstrate that one 30-minute application of hot air has the potential to eradicate head lice infestations. In summary, hot air is an effective, safe treatment and one to which lice are unlikely to evolve resistance. PMID- 17079568 TI - Circumcision status and risk of sexually transmitted infection in young adult males: an analysis of a longitudinal birth cohort. AB - OBJECTIVES: Previous research suggests that male circumcision may be a protective factor against the acquisition of sexually transmitted infections; however, studies examining this question have produced mixed results. The aim of this study was to examine the association between circumcision status and sexually transmitted infection risk using a longitudinal birth cohort study. METHODS: Data were gathered as part of the Christchurch Health and Development Study, a 25-year longitudinal study of a birth cohort of New Zealand children. Information was obtained on: (1) the circumcision status of males in the cohort before 15 years old, (2) measures of self-reported sexually transmitted infection from ages 18 to 25 years, and (3) childhood, family, and related covariate factors. RESULTS: Being uncircumcised had a statistically significant bivariate association with self-reported sexually transmitted infection. Adjustment for potentially confounding factors, including number of sexual partners and unprotected sex, as well as background and family factors related to circumcision, did not reduce the association between circumcision status and reports of sexually transmitted infection. Estimates of the population-attributable risk suggested that universal neonatal circumcision would have reduced rates of sexually transmitted infection in this cohort by 48.2%. CONCLUSIONS: These findings suggest that uncircumcised males are at greater risk of acquiring sexually transmitted infection than circumcised males. Male circumcision may reduce the risk of sexually transmitted infection acquisition and transmission by up to one half, suggesting substantial benefits accruing from routine neonatal circumcision. PMID- 17079569 TI - School bus-related injuries among children and teenagers in the United States, 2001-2003. AB - OBJECTIVE: The purpose of this work was to describe the epidemiology of nonfatal school bus-related injuries among children and teenagers aged < or = 19 years in the United States. DESIGN/METHODS: Nationally representative data from the National Electronic Injury Surveillance System All-Injury Program operated by the US Consumer Product Safety Commission were analyzed. Case subjects included all of the patients in the National Electronic Injury Surveillance System All-Injury Program database who were treated in a hospital emergency department for a nonfatal school bus-related injury from 2001 to 2003. RESULTS: There were an estimated 51,100 school bus-related injuries treated in US emergency departments from 2001 to 2003, for a national estimate of 17,000 injuries (rate: 21.0 per 100,000 population) annually. Ninety-seven percent of children were treated and released from the hospital. Children 10 to 14 years of age accounted for the greatest proportion of injuries (43.0%; rate: 34.7) compared with all other age groups. Motor vehicle crashes accounted for 42.3% of all injuries, followed by injuries that occurred as the child was boarding/alighting/approaching the bus (23.8%). Head injuries accounted for more than half (52.1%) of all injuries among children < 10 years of age, whereas lower extremity injuries predominated among children 10 to 19 years of age (25.5%). Strains and sprains accounted for the highest percentage of all injuries, followed by contusions and abrasions (28.3%) and lacerations (14.9%). More than three quarters (77.7%) of lacerations were to the head. CONCLUSIONS: This is the first study to describe nonfatal school bus related injuries to US children and teenagers treated in US hospital emergency departments using a national sample. This study identified a much greater annual number of school bus-related injuries to children than reported previously. PMID- 17079570 TI - Psychological problems in children with daytime wetting. AB - OBJECTIVE: This population-based study investigated the psychological problems associated with daytime wetting in children. METHODS: A sample of 8213 children (age range: 7 years 6 months to 9 years 3 months) who were enrolled in the population-based Avon Longitudinal Study of Parents and Children participated in this study. Parents completed a postal questionnaire asking about their children's toileting behavior and assessing psychological problems, including childhood emotional and behavioral problems (99% completed the questionnaire by the time their child was 8 years 3 months of age). The rate of psychological problems was compared in children with daytime wetting and in those with no daytime wetting. Analyses adjusted for developmental delay, gender, sociodemographic background, stressful life events, and soiling. RESULTS: Chi2 tests of association and multivariable logistic regression indicate that children with daytime wetting have a higher rate of parent-reported psychological problems than children who have no daytime wetting. It is particularly notable that the reported rates of attention and activity problems, oppositional behavior, and conduct problems in daytime wetting children were around twice the rates reported in children with no daytime wetting. CONCLUSIONS: The increased vulnerability to psychological problems in children as young as 7 years of age with daytime wetting highlights the importance of parents seeking early intervention for the condition to help prevent later psychological problems. Although treatment in a pediatric setting is often successful, clinicians should be aware of the increased risk of disorders, such as attention-deficit/hyperactivity disorder, in children with daytime wetting, because this is likely to interfere with treatment. PMID- 17079571 TI - Cost-outcome analysis of booster seats for auto occupants aged 4 to 7 years. AB - OBJECTIVES: The purpose of this work was to analyze the societal return on investment in booster seats and in laws requiring their use in the United States. Booster seats reduce crash-related injury. Their use is mandatory for vehicle occupants aged 4 to 7 years in most of the United States. This study estimates the injury cost savings attributable to booster seat use. METHODS: Seat cost came from pricing on the Web and at retailers. Costs of passing and enforcing a legal mandate were estimated as a percentage of the costs of seat use. Injury risk when belted absent a seat was computed from national probability samples of crashes in the last years before booster seats entered into general use (1993-1999). Published estimates were used of the percentage of reduction in injuries achieved with booster seats, the mix of diagnoses reduced, and injury cost by diagnosis. The computations used a 3% discount rate. We studied the net cost per quality adjusted life year saved, benefit-cost ratio, and net savings per seat. RESULTS: A booster seat costs 30 dollars plus 167 dollars for maintenance and time spent on installation and use. This investment saves 1854 dollars per seat, a return on investment of 9.4 to 1. Even lower bound estimates in sensitivity analysis indicated that society would benefit from the use of booster seats. Seat laws offer a return of 8.6 to 1. CONCLUSIONS: Belt-positioning booster seats offer a sound return on investment. Booster seat use laws should be passed, publicized, and enforced nationwide. PMID- 17079572 TI - Doppler flow velocimetry in the superior mesenteric artery on the first day of life in preterm infants and the risk of neonatal necrotizing enterocolitis. AB - OBJECTIVE: The purpose of this work was to relate Doppler indices of splanchnic perfusion and vascular resistance to the risk of developing necrotizing enterocolitis. METHODS: We conducted a prospective cohort study with analysis of Doppler flow velocity waveforms of splanchnic vessels on the first day of life. Clinical management and diagnosis of necrotizing enterocolitis were performed blind to the Doppler results in a tertiary NICU on 64 eligible preterm neonates admitted for intensive care. We measured necrotizing enterocolitis using an objective diagnostic classification. RESULTS: When adjusted for gestational age at birth, the following indices of the Doppler flow velocity wave form in the superior mesenteric artery were significantly predictive of the risk of necrotizing enterocolitis: end-diastolic velocity, mean velocity, and pulsatility index. The association between necrotizing enterocolitis and Doppler velocimetry indicative of high vascular resistance was independent of a range of other factors and comorbidities (race, mode of delivery, umbilical arterial catheter, growth restriction, patent ductus arteriosus, jaundice, respiratory distress syndrome, mechanical ventilation, and hypotension). CONCLUSIONS: We concluded that neonates with high resistance patterns of blood flow velocity in the superior mesenteric artery on the first day of life are at increased risk of developing necrotizing enterocolitis. PMID- 17079573 TI - Effects of a combined therapy of erythropoietin, iron, folate, and vitamin B12 on the transfusion requirements of extremely low birth weight infants. AB - OBJECTIVES: Erythropoietin is frequently administered to premature infants to stimulate erythropoiesis. The primary goal of erythropoietin therapy is to reduce transfusions, but the efficacy of erythropoietin has not been convincingly demonstrated in this regard. The aim of this trial was to investigate whether combined administration of vitamin B12, folic acid, iron, and erythropoietin could decrease transfusion requirements in extremely low birth weight infants. PATIENTS AND METHODS: In a randomized, controlled trial, extremely low birth weight infants with a birth weight < or = 800 g and a gestational age < or = 32 weeks were randomly assigned to a group receiving combination treatment or a control arm. RESULTS: The treatment increased levels of folate in red blood cells, vitamin B12, ferritin, transferrin receptor levels in plasma, and reticulocyte counts. The proportion of infants requiring no transfusions was lower in the treatment group (38%) as compared with controls (5%). The treatment group and the need for mechanical ventilation were independent predictors of the number of transfusions in multiple regression analysis. Cox regression analysis indicated that combined therapy resulted in a 79% risk reduction for any transfusion. CONCLUSION: Combined treatment with erythropoietin, intravenous iron, folate, and vitamin B12 during the first weeks reduces the need for transfusion in extremely low birth weight infants. PMID- 17079575 TI - A systematic review for the effects of television viewing by infants and preschoolers. AB - OBJECTIVE: The purpose of this work was to conduct a systematic review of experimental trials for the effects of television viewing by infants and preschoolers. METHODS: Searches were conducted as of January 2005 on several online bibliographic databases for the disciplines of medicine, psychology, education, and communications. Relevant bibliographies were also reviewed. The article contents were critically evaluated to determine whether they met inclusion criteria. Data were extracted from each included article regarding study methods and results. RESULTS: Twelve controlled trials met all of the criteria for inclusion in this review. All of the studies focused on the effect of television content rather than viewing time. The findings suggest that educational television programs are successful in broadening young children's knowledge, affecting their racial attitudes, and increasing their imaginativeness. There is insufficient experimental evidence for effects of viewing these programs on either children's prosocial behavior or their aggressive behavior. Finally, there is some evidence that viewing cartoons has a negative effect on children's attentional abilities. CONCLUSIONS: A number of gaps remain in the literature. Although the amount of time children spend viewing television raises concern, the studies presented here focus only on content. Despite the fact that infants are the fastest growing television market segment, the controlled trials only include children aged > or = 3 years. Finally, the clinical utility of the findings are questionable given that most studies included small group sizes, all took place in nonnaturalistic settings, and all but 1 study only evaluated short-term effects of television viewing. PMID- 17079574 TI - Diffusion tensor imaging of frontal white matter and executive functioning in cocaine-exposed children. AB - BACKGROUND: Although animal studies have demonstrated frontal white matter and behavioral changes resulting from prenatal cocaine exposure, no human studies have associated neuropsychological deficits in attention and inhibition with brain structure. We used diffusion tensor imaging to investigate frontal white matter integrity and executive functioning in cocaine-exposed children. METHODS: Six direction diffusion tensor images were acquired using a Siemens 3T scanner with a spin-echo echo-planar imaging pulse sequence on right-handed cocaine exposed (n = 28) and sociodemographically similar non-exposed children (n = 25; mean age: 10.6 years) drawn from a prospective, longitudinal study. Average diffusion and fractional anisotropy were measured in the left and right frontal callosal and frontal projection fibers. Executive functioning was assessed using two well-validated neuropsychological tests (Stroop color-word test and Trail Making Test). RESULTS: Cocaine-exposed children showed significantly higher average diffusion in the left frontal callosal and right frontal projection fibers. Cocaine-exposed children were also significantly slower on a visual-motor set-shifting task with a trend toward lower scores on a verbal inhibition task. Controlling for gender and intelligence, average diffusion in the left frontal callosal fibers was related to prenatal exposure to alcohol and marijuana and an interaction between cocaine and marijuana exposure. Performance on the visual motor set-shifting task was related to prenatal cocaine exposure and an interaction between cocaine and tobacco exposure. Significant correlations were found between test performance and fractional anisotropy in areas of the frontal white matter. CONCLUSIONS: Prenatal cocaine exposure, alone and in combination with exposure to other drugs, is associated with slightly poorer executive functioning and subtle microstructural changes suggesting less mature development of frontal white matter pathways. The relative contribution of postnatal environmental factors, including characteristics of the caregiving environment and stressors associated with poverty and out-of-home placement, on brain development and behavioral functioning in polydrug-exposed children awaits further research. PMID- 17079576 TI - Compliance with the recommendations for 2 doses of trivalent inactivated influenza vaccine in children less than 9 years of age receiving influenza vaccine for the first time: a Vaccine Safety Datalink study. AB - OBJECTIVES: Children <9 years of age do not respond optimally to a first dose of trivalent inactivated influenza vaccine, and so 2 doses of trivalent inactivated influenza vaccine are recommended for children <9 years of age who are being vaccinated for the first time. We conducted a population-based retrospective cohort study to evaluate compliance with the 2-dose trivalent inactivated influenza vaccine recommendations. POPULATION AND SETTING: We evaluated 125,928 children 6 months through 8 years of age who were enrolled in health maintenance organizations in the United States participating in the Vaccine Safety Datalink project and who received their first dose of trivalent inactivated influenza vaccine in the 2001-2002, 2002-2003, or 2003-2004 influenza seasons. RESULTS: Compliance with the 2 dose recommendations varied by age group and influenza season. Among children 6 to 23 months of age, the proportion of first-vaccinated children who received a second vaccination was 44% in 2001-2002, 54% in 2002 2003, and 29% in 2003-2004. Among children 2 to 8 years of age, the corresponding proportions were 15%, 24%, and 12%, respectively. In all seasons, compliance with the second vaccination was highest in children first vaccinated by mid-November. CONCLUSIONS: The majority of children who received their first dose of trivalent inactivated influenza vaccine did not complete the 2-dose series. The recently expanded recommendation for universal vaccination of children 6 to 59 months of age and their household contacts will substantially increase the number of children targeted for a first influenza vaccination. Noncompliance with the 2 dose trivalent inactivated influenza vaccine series may be associated with suboptimal protection against infection, which may impact the magnitude of the direct and indirect benefits achieved by the vaccination program. PMID- 17079577 TI - Delayed extubation to nasal continuous positive airway pressure in the immature baboon model of bronchopulmonary dysplasia: lung clinical and pathological findings. AB - OBJECTIVE: Using the 125-day baboon model of bronchopulmonary dysplasia treated with prenatal steroid and exogenous surfactant, we hypothesized that a delay of extubation from low tidal volume positive pressure ventilation to nasal continuous positive airway pressure at 5 days (delayed nasal continuous positive airway pressure group) would not induce more lung injury when compared with baboons aggressively weaned to nasal continuous positive airway pressure at 24 hours (early nasal continuous positive airway pressure group), because both received positive pressure ventilation. METHODS AND RESULTS: After delivery by cesarean section at 125 days (term: 185 days), infants received 2 doses of Curosurf (Chiesi Farmaceutica S.p.A., Parma, Italy) and daily caffeine citrate. The delay in extubation to 5 days resulted in baboons in the delayed nasal continuous positive airway pressure group having a lower arterial to alveolar oxygen ratio, high PaCO2, and worse respiratory function. The animals in the delayed nasal continuous positive airway pressure group exhibited a poor respiratory drive that contributed to more reintubations and time on mechanical ventilation. A few animals in both groups developed necrotizing enterocolitis and/or sepsis, but infectious pneumonias were not documented. Cellular bronchiolitis and peribronchiolar alveolar wall thickening were more frequently seen in the delayed nasal continuous positive airway pressure group. Bronchoalveolar lavage levels of interleukin-6, interleukin-8, monocyte chemotactic protein-1, macrophage inflammatory protein-1 alpha, and growth regulated oncogene-alpha were significantly increased in the delayed nasal continuous positive airway pressure group. Standard and digital morphometric analyses showed no significant differences in internal surface area and nodal measurements between the groups. Platelet endothelial cell adhesion molecule vascular staining was not significantly different between the 2 nasal continuous positive airway pressure groups. CONCLUSIONS: Volutrauma and/or low-grade colonization of airways secondary to increased reintubations and ventilation times are speculated to play causative roles in the delayed nasal continuous positive airway pressure group findings. PMID- 17079578 TI - Sleep environment, positional, lifestyle, and demographic characteristics associated with bed sharing in sudden infant death syndrome cases: a population based study. AB - BACKGROUND: In 2005, the American Academy of Pediatrics Task Force on Sudden Infant Death Syndrome recommended that infants not bed share during sleep. OBJECTIVE: Our goal was to characterize the profile of risk factors associated with bed sharing in sudden infant death syndrome cases. DESIGN/METHODS: We conducted a population-based retrospective review of sudden infant death syndrome cases in New Jersey (1996-2000) dichotomized by bed-sharing status and compared demographic, lifestyle, bedding-environment, and sleep-position status. RESULTS: Bed-sharing status was reported in 239 of 251 cases, with sharing in 39%. Bed sharing cases had a higher percentage of bedding risks (44.1% vs 24.7%), exposure to bedding risks in infants discovered prone (57.1% vs 28.2%), and lateral sleep placement (28.9% vs 17.8%). The prone position was more common for bed-sharing and non-bed-sharing cases at placement (45.8% and 51.1%, respectively) and discovery (59.0% and 64.4%, respectively). In multivariable logistic-regression analyses, black race, mother <19 years, gravida >2, and maternal smoking were associated with bed sharing. There was a trend toward less breastfeeding in bed sharing cases (22% vs 35%). In bed-sharing cases, those breastfed were younger than those who were not and somewhat more exposed to bedding risks (64.7% vs 45.1%) but less likely to be placed prone (11.8% vs 52.9%) or have maternal smoking (33% vs 66%). CONCLUSIONS: Bed-sharing cases were more likely to have had bedding-environment and sleep-position risks and higher ratios of demographic and lifestyle risk factors. Bed-sharing subjects who breastfed had a risk profile distinct from those who were not breastfed cases. Risk and situational profiles can be used to identify families in greater need of early guidance and to prepare educational content to promote safe sleep. PMID- 17079579 TI - Neonatal hemochromatosis: long-term experience with favorable outcome. AB - OBJECTIVE: Neonatal hemochromatosis is a severe, often fatal multiorgan disorder of iron metabolism. Liver transplantation can be curative; the benefit of antioxidant treatment is discussed controversially. We summarize our experience with neonatal hemochromatosis over the past 13 years. METHODS: A retrospective study was performed of 16 patients with acute liver failure attributable to neonatal hemochromatosis between 1992 and 2004. RESULTS: Median age at the onset of neonatal hemochromatosis was 2 days (range: 0-21 days). Median weight at the time of diagnosis was 2900 g (range: 1520-4200 g). All patients had elevated ferritin levels (median: 4179 microg/L), and transferrin saturation (median: 99%). Fourteen patients (87.5%) showed significant hepatocyte siderosis in biopsies; 4 children had additional iron deposition in extrahepatic tissue. Four patients were diagnosed by MRI. Seven infants received liver transplants, 5 of them in combination with a preceding antioxidant treatment. Four children (25%) received antioxidants without the necessity of liver transplantation and were in good clinical condition at the time of this evaluation. Five patients (31.3%) died, 3 of them without any treatment because of initial fulminant multiorgan failure. In September 2005, 68.7% of the patients were still alive after a median follow-up of 5 years. CONCLUSIONS: Neonatal hemochromatosis is a severe metabolic disease, but early antioxidant treatment and liver transplantation in addition to optimal medical care can improve the outcome dramatically. Children with moderate liver failure can survive without liver transplantation, but should be monitored closely for deterioration. PMID- 17079580 TI - Fruit juice intake predicts increased adiposity gain in children from low-income families: weight status-by-environment interaction. AB - OBJECTIVE: Our goal was to test the hypothesis that increased fruit juice intake and parental restriction of children's eating are associated with increased adiposity gain and whether exposure to nutritional counseling predicted reduced adiposity gain among children. PATIENTS AND METHODS: A sample of parents or guardians of children aged 1 to 4 years who attended 1 of 49 Special Supplemental Nutrition Program for Women, Infants, and Children agencies in New York State were surveyed in 1999 or 2000 (N = 2801). The survey addressed children's dietary intake, parental feeding practices, and parental exposure to nutritional counseling messages to increase fruit, vegetable, and low-fat milk intakes. Each child's height and weight were measured approximately every 6 months for up to 48 months. A prospective cohort design was used in which survey variables were the predictors and the outcome was change in children's adiposity, defined as change in age- and gender-standardized BMI per month (ie, BMI z-score slope). RESULTS: Controlling for gender and ethnicity, the relationship between juice intake and adiposity gain depended on children's initial overweight status. Among children who were initially either at risk for overweight or overweight, increased fruit juice intake was associated with excess adiposity gain, whereas parental offerings of whole fruits were associated with reduced adiposity gain. Each additional daily serving of fruit juice was associated with an excess adiposity gain of 0.009 SD per month. Feeding restriction was greater among parents whose children were initially at risk for overweight or overweight compared with those at a healthy weight. Parental exposure to nutritional messages was not associated with reduced child adiposity gain. CONCLUSION: This study supports the Institute of Medicine recommendations to reduce fruit juice intake as a strategy for overweight prevention in high-risk children. PMID- 17079581 TI - Improved outcomes of extremely premature outborn infants: effects of strategic changes in perinatal and retrieval services. AB - OBJECTIVE: The goal was to evaluate the impact of statewide coordinated changes in perinatal support and retrieval services on the outcomes of extremely premature births occurring outside perinatal centers in the state of New South Wales, Australia. METHODS: The intervention included additional, network coordinated, perinatal telephone advice to optimize in utero transfers and centralization of the neonatal retrieval system, with preferential admission of retrieved infants (outborn infants) to perinatal centers instead of freestanding pediatric hospitals, from the middle of 1995. Population birth and NICU admission cohorts of infants of 23 to 28 weeks of gestation were studied. Outcomes of epoch 1 (1992 to the middle of 1995; 1778 births and 1100 NICU admissions) were compared with those of epoch 2 (1997-2002; 3099 births and 2100 NICU admissions), after an 18-month washout period. RESULTS: There were 25% fewer nontertiary hospital live births (19.7% vs 14.9%) and more prenatal steroid use. Despite an 11.4% average annual increase in NICU admissions between the 2 epochs, fewer infants were outborn (12.0% vs 9.3%) and outborn mortality rates decreased significantly (39.4% vs 25.1%), particularly for those between 27 and 28 weeks of gestation. The overall improvement was equivalent to 1 extra survivor per 16 New South Wales births. There were also significantly fewer serious outcome morbidities in outborn infants during epoch 2, over the improvements in inborn infants. CONCLUSIONS: Statewide coordinated strategies in reducing nontertiary hospital births and optimizing transport of outborn infants to perinatal centers have improved considerably the outcomes of extremely premature infants. These findings have vital implications for health outcomes and resource planning. PMID- 17079582 TI - Predicting outcomes of neonates diagnosed with hypoxemic-ischemic encephalopathy. AB - OBJECTIVE: The goals were to identify predictor variables and to develop scoring systems and classification trees to predict death/disability or death in infants with hypoxic-ischemic encephalopathy. METHODS: Secondary analysis of data from the multicenter, randomized, controlled, National Institute of Child Health and Human Development Neonatal Research Network trial of hypothermia in hypoxic ischemic encephalopathy was performed. Data for 205 neonates diagnosed as having hypoxic-ischemic encephalopathy were studied. Logistic regression analysis was performed by using clinical and laboratory variables available within 6 hours of birth, with death or moderate/severe disability at 18 to 22 months or death as the outcomes. By using the identified variables and odds ratios, scoring systems to predict death/disability or death were developed, weighting each predictor in proportion to its odds ratio. In addition, classification and regression tree analysis was performed, with recursive partitioning and automatic selection of optimal cutoff points for variables. Correct classification rates for the scoring systems, classification and regression tree models, and early neurologic examination were compared. RESULTS: Correct classification rates were 78% for death/disability and 71% for death with the scoring systems, 80% and 77%, respectively, with the classification and regression tree models, and 67% and 73% with severe encephalopathy in early neurologic examination. Correct classification rates were similar in the hypothermia and control groups. CONCLUSIONS: Among neonates diagnosed as having hypoxic-ischemic encephalopathy, the classification and regression tree model, but not the scoring system, was superior to early neurologic examination in predicting death/disability. The 3 models were comparable in predicting death. Only a few components of the early neurologic examination were associated with poor outcomes. These scoring systems and classification trees, if validated, may help in assessments of prognosis and may prove useful for risk-stratification of infants with hypoxic-ischemic encephalopathy for clinical trials. PMID- 17079583 TI - The potential impact of poison control centers on rural hospitalization rates for poisoning. AB - OBJECTIVE: This study tested the hypothesis that underutilization of poison control centers is associated with increased rates of hospitalizations attributable to poisonings in rural areas. METHODS: To measure the potential impact of poison control centers on hospitalization rates in rural areas among people who visit emergency departments because of poisoning, we estimated the reduction in hospitalization rates associated with increased rates of calls to centers. We used the 2003 State Inpatient Database and State Emergency Department Database from the Healthcare Cost and Utilization Project to calculate the numbers of emergency department visits and hospitalizations for each county in the 12 states analyzed. We used Toxic Exposure Surveillance System data from the American Association of Poison Control Centers to calculate the number of human exposure calls per capita according to county. RESULTS: In rural counties, a 1% higher poison control center human poison exposure call rate was associated with a 0.19% lower hospitalization rate among people who visited emergency departments because of poisoning. If the observed association is causative, then 43.3 calls would prevent 1 hospital admission, yielding 7321 dollars in net cost savings and a return on investment of 5.9:1 (from the health care system perspective). CONCLUSIONS: Our results establish the existence of the hypothesized association between rural poison control center utilization rates and hospitalization rates among emergency department-treated poisoning patients. PMID- 17079584 TI - Outcome at 6 months after admission for pediatric intensive care: a report of a national study of pediatric intensive care units in the United kingdom. AB - OBJECTIVE: The goal was to measure, by using the Health Utilities Index, the health status of children 6 months after admission to PICUs in the United Kingdom. METHODS: All PICUs in the United Kingdom were invited to participate. Children who were > or = 6 months of age at admission and were discharged alive from participating units during a 1-year period were eligible for this study. Children with completed consent forms who had survived to 6 months after admission received the Health Utilities Index questionnaire. RESULTS: Admission data on 7214 admissions to 22 units were collected between March 2001 and February 2002. Of those patients, 6786 survived to unit discharge, and consent for follow-up contact was obtained for 2642. At 6 months after admission, the mortality rate was estimated at 11.1% and 2034 children were still alive and contactable. Of those children, Health Utilities Index questionnaires were returned by 1455. No impairment was indicated for 767, 951, 940, 919, 962, and 939 children with respect to the Health Utilities Index sensation, cognition, emotion, pain, mobility, and self-care attributes, respectively. The mean +/- SE Health Utilities Index utility score was 0.73 +/- 0.01, with 397 children (27.3%) in full health. CONCLUSIONS: Death after pediatric intensive care in the United Kingdom is uncommon, making assessment of health status important. At 6 months after admission, there is significant morbidity. To assess the impact of strategies to improve or to optimize care on longer-term outcomes, standardized collection of data on preexisting comorbidities and illness severity is required. PMID- 17079585 TI - Validity of parentally reported weight and height for preschool-aged children in Belgium and its impact on classification into body mass index categories. AB - OBJECTIVE: The aim of this study was to evaluate the validity of parentally reported weight and height values for preschool-aged children (3-7 years of age) in Belgium and the accuracy of BMI classifications using those data. METHODS: The subjects were 297 preschool-aged children (157 boys and 140 girls). A questionnaire with questions about height and weight was distributed to the children and completed by their parents. Nurses of the Centres for Pupils Counselling conducted anthropometric measurements by following standardized procedures. National and international age- and gender-specific BMI cutoff values were used to determine BMI categories. RESULTS: Mean differences between reported and measured values for weight (-0.57 kg) and BMI (-0.51 kg/m2) were statistically significant, although small. For height, the mean difference (0.14 cm) was not statistically significant. When national and international BMI cutoff values for overweight were used, sensitivity values were 47% and 44%, respectively, and specificity values were 94% and 95%. Among all children requiring nutritional advice on the basis of being overweight or obese, more than one half of the overweight children and >75% of the obese children would be missed with the use of parentally reported weight and height values. Conversely, 70% of the children being classified as underweight with the use of these reported data could be encouraged wrongly to gain weight. The bias of parent reported BMI values was significantly greater when weight and height were both guessed, rather than being measured at home. CONCLUSIONS: Our results show the inaccuracy of parentally reported weight and height values in Belgium for classifying preschool-aged children into BMI categories. Therefore, accurate measurements of weight and height should be encouraged in studies in which BMI of children is a variable of interest. PMID- 17079586 TI - Acute pediatric rhabdomyolysis: causes and rates of renal failure. AB - OBJECTIVES: The goals were to (1) compare the causes, clinical presentation, and prevalence of acute renal failure in pediatric rhabdomyolysis with the published data for adults; (2) determine predictors of acute renal failure in pediatric patients with rhabdomyolysis; and (3) explore the relationship of acute renal failure with treatment modalities such as fluid and bicarbonate administration. METHODS: We performed a retrospective chart review to identify patients with creatinine kinase levels of > 1000 IU/L who were treated in the emergency department of a tertiary pediatric hospital between 1993 and 2003, and we constructed regression models. RESULTS: Two hundred ten patients were studied. One hundred ninety-one patients met study eligibility (128 male and 63 female), with a median age of 11 years. The most common documented symptoms were muscle pain (45%), fever (40%), and symptoms of viral infection (39%). The most common causes of pediatric rhabdomyolysis were viral myositis (38%), trauma (26%), and connective tissue disease (5%). Six of 37 patients with creatinine kinase levels of > or = 6000 IU/L had previously undiagnosed dermatomyositis or hereditary metabolic disease, compared with 10 of 154 patients with creatinine kinase levels of 1000 to 5999 IU/L. Nine of 191 patients developed acute renal failure. None of 99 patients with initial urinary heme dipstick results of < 2+ developed acute renal failure, compared with 9 of 44 patients with urinary heme dipstick results of > or = 2+. Higher initial creatinine kinase levels and higher fluid administration rates were associated with higher maximal creatinine levels. CONCLUSIONS: The cause of acute pediatric rhabdomyolysis is different from that of adult rhabdomyolysis. The risk of acute renal failure in children is much less than the risk reported for adults. PMID- 17079587 TI - Exercise therapy as a treatment for psychopathologic conditions in obese and morbidly obese adolescents: a randomized, controlled trial. AB - OBJECTIVE: We conducted a proof-of-concept, randomized, controlled trial to investigate the effects of a supervised exercise therapy intervention on psychopathologic outcomes in obese adolescents. METHODS: The participant sample consisted of 81 adolescents (age: 11-16 years) who had been referred to a children's hospital for evaluation of obesity or who responded to a community advertisement. Participants were assigned randomly to exercise therapy, an equal contact exercise placebo intervention, or usual care. Intervention participants attended 3 one-on-one sessions per week for 8 weeks and then completed a home program for another 6 weeks. Outcomes included self-perceptions (self-esteem), depression, affect, physical activity, aerobic fitness, and BMI. RESULTS: A total of 18 of 81 participants were categorized as morbidly obese (BMI SD score: > 3.5; adult equivalent BMI: > or = 40). At baseline, 30.3% of participants had a Children's Depression Inventory score of > or = 13, and 27% reported recent suicidal ideation. Repeated-measures mixed analysis of covariance (controlling for baseline scores) revealed significant changes in physical self-worth, associated measures of self-esteem, and physical activity over time, consistently favoring exercise therapy. There were no significant changes in BMI. CONCLUSIONS: Findings confirmed psychopathologic conditions as a serious health concern in obese and morbidly obese adolescents. Our study is the first randomized, controlled trial to demonstrate that a brief supervised exercise therapy intervention has the potential to improve psychopathologic outcomes significantly and to increase physical activity in obese adolescents, relative to usual care. PMID- 17079588 TI - Comparison of the immunogenicity and reactogenicity of a prophylactic quadrivalent human papillomavirus (types 6, 11, 16, and 18) L1 virus-like particle vaccine in male and female adolescents and young adult women. AB - OBJECTIVE: Prophylactic vaccination of 16- to 23-year-old females with a quadrivalent human papillomavirus (types 6, 11, 16, 18) L1 virus-like particle vaccine has been shown to prevent type-specific human papillomavirus infection and associated clinical disease. We conducted a noninferiority immunogenicity study to bridge the efficacy findings in young women to preadolescent and adolescent girls and boys, who represent a primary target for human papillomavirus vaccination. METHODS: We enrolled 506 girls and 510 boys (10-15 years of age) and 513 females (16-23 years of age). Participants were vaccinated on day 1, at month 2, and at month 6, and serology testing was performed on day 1 and at months 3 and 7 on blinded samples. Neutralizing antibody concentrations were determined using type-specific immunoassays and summarized as geometric mean titers and seroconversion rates. Vaccine tolerability also was assessed. RESULTS: By month 7, seroconversion rates were > or = 99% for all 4 human papillomavirus types in each group. By month 7, compared with women, anti-human papilloma virus geometric mean titers in girls or boys were noninferior and were 1.7- to 2.7-fold higher. Most (> 97%) injection-site adverse events were mild to moderate in intensity. Significantly more boys (13.8%) and girls (12.8%) than women (7.3%) reported fevers > or = 37.8 degrees C within 5 days of vaccination. Most (96.4%) fevers were mild (< 39 degrees C). CONCLUSIONS: Noninferior immunogenic responses to all 4 human papillomavirus types in the quadrivalent vaccine permit the bridging of efficacy data that were generated in young women to girls. The results in boys lend support for the implementation of gender-neutral human papillomavirus vaccination programs. This vaccine generally was well tolerated. PMID- 17079589 TI - Pediatric cyanide poisoning: causes, manifestations, management, and unmet needs. AB - Confirmed cases of childhood exposure to cyanide are rare despite multiple potential sources including inhalation of fire smoke, ingestion of toxic household and workplace substances, and ingestion of cyanogenic foods. Because of its infrequent occurrence, medical professionals may have difficulty recognizing cyanide poisoning, confirming its presence, and treating it in pediatric patients. The sources and manifestations of acute cyanide poisoning seem to be qualitatively similar between children and adults, but children may be more vulnerable than adults to poisoning from some sources. The only currently available antidote in the United States (the cyanide antidote kit) has been used successfully in children but has particular risks associated with its use in pediatric patients. Because hemoglobin kinetics vary with age, methemoglobinemia associated with nitrite-based antidotes may be excessive at standard adult dosing in children. A cyanide antidote with a better risk/benefit ratio than the current agent available in the United States is desirable. The vitamin B12 precursor hydroxocobalamin, which has been used in Europe, may prove to be an attractive alternative to the cyanide antidote kit for pediatric patients. In this article we review the available data on the sources, manifestations, and treatment of acute cyanide poisoning in children and discuss unmet needs in the management of pediatric cyanide poisoning. PMID- 17079591 TI - Counseling about firearms: proposed legislation is a threat to physicians and their patients. AB - In early 2006, 2 separate but virtually identical bills were introduced in the Virginia and West Virginia legislatures that would have profoundly affected the relationship between a physician and his or her patients. Each bill would have prohibited a physician from asking a patient if he or she owned firearms for the purpose of counseling that patient about ways to reduce risks associated with firearms. Penalties for violation of the bills included revocation of a physician's license to practice. The Virginia bill was initially approved by its state House of Delegates by a vote of 88 to 11. It was ultimately defeated in a Virginia Senate committee. The West Virginia bill did not receive a vote during the 2006 legislative session. Although neither bill became law this year, this type of bill is likely to reappear in future legislative sessions. The Virginia and West Virginia bills were contrary to the best-practices recommendations of medical societies, including the American Academy of Pediatrics. Anticipatory guidance regarding firearms can indeed reduce risks to patients. Yet, the bills would have preferred the judgment of legislators over physicians regarding this aspect of the practice of medicine. In addition, the 2 bills raise legal issues regarding both medical malpractice and the First Amendment protection of the freedom of speech. The Virginia and West Virginia bills would have treated risks associated with firearms differently from other hazards and interfered with a physician's ability to protect his or her patients. The Virginia bill was defeated, in part, through the efforts of physicians to educate legislators. However, physicians must remain prepared to respond to similar state legislative initiatives in the future. PMID- 17079590 TI - Definition and classification of negative motor signs in childhood. AB - In this report we describe the outcome of a consensus meeting that occurred at the National Institutes of Health in Bethesda, Maryland, March 12 through 14, 2005. The meeting brought together 39 specialists from multiple clinical and research disciplines including developmental pediatrics, neurology, neurosurgery, orthopedic surgery, physical therapy, occupational therapy, physical medicine and rehabilitation, neurophysiology, muscle physiology, motor control, and biomechanics. The purpose of the meeting was to establish terminology and definitions for 4 aspects of motor disorders that occur in children: weakness, reduced selective motor control, ataxia, and deficits of praxis. The purpose of the definitions is to assist communication between clinicians, select homogeneous groups of children for clinical research trials, facilitate the development of rating scales to assess improvement or deterioration with time, and eventually to better match individual children with specific therapies. "Weakness" is defined as the inability to generate normal voluntary force in a muscle or normal voluntary torque about a joint. "Reduced selective motor control" is defined as the impaired ability to isolate the activation of muscles in a selected pattern in response to demands of a voluntary posture or movement. "Ataxia" is defined as an inability to generate a normal or expected voluntary movement trajectory that cannot be attributed to weakness or involuntary muscle activity about the affected joints. "Apraxia" is defined as an impairment in the ability to accomplish previously learned and performed complex motor actions that is not explained by ataxia, reduced selective motor control, weakness, or involuntary motor activity. "Developmental dyspraxia" is defined as a failure to have ever acquired the ability to perform age-appropriate complex motor actions that is not explained by the presence of inadequate demonstration or practice, ataxia, reduced selective motor control, weakness, or involuntary motor activity. PMID- 17079592 TI - The National Children's Study: a 21-year prospective study of 100,000 American children. AB - Prospective, multiyear epidemiologic studies have proven to be highly effective in discovering preventable risk factors for chronic disease. Investigations such as the Framingham Heart Study have produced blueprints for disease prevention and saved millions of lives and billions of dollars. To discover preventable environmental risk factors for disease in children, the US Congress directed the National Institute of Child Health and Human Development, through the Children's Health Act of 2000, to conduct the National Children's Study. The National Children's Study is hypothesis-driven and will seek information on environmental risks and individual susceptibility factors for asthma, birth defects, dyslexia, attention-deficit/hyperactivity disorder, autism, schizophrenia, and obesity, as well as for adverse birth outcomes. It will be conducted in a nationally representative, prospective cohort of 100,000 US-born children. Children will be followed from conception to 21 years of age. Environmental exposures (chemical, physical, biological, and psychosocial) will be assessed repeatedly during pregnancy and throughout childhood in children's homes, schools, and communities. Chemical assays will be performed by the Centers for Disease Control and Prevention, and banks of biological and environmental samples will be established for future analyses. Genetic material will be collected on each mother and child and banked to permit study of gene-environment interactions. Recruitment is scheduled to begin in 2007 at 7 Vanguard Sites and will extend to 105 sites across the United States. The National Children's Study will generate multiple satellite studies that explore methodologic issues, etiologic questions, and potential interventions. It will provide training for the next generation of researchers and practitioners in environmental pediatrics and will link to planned and ongoing prospective birth cohort studies in other nations. Data from the National Children's Study will guide development of a comprehensive blueprint for disease prevention in children. PMID- 17079593 TI - A promising opportunity for developmental and behavioral pediatrics at the interface of neuroscience, psychology, and social policy: remarks on receiving the 2005 C. Anderson Aldrich Award. PMID- 17079594 TI - Daubert opinion requires judges to screen scientific evidence. PMID- 17079595 TI - A microscopic view of medical care and its costs. PMID- 17079596 TI - The Daubert decision. PMID- 17079597 TI - Hypocalcemic seizures and secondary bilateral femoral fractures in an adolescent with primary vitamin D deficiency. AB - Nutritional rickets and osteomalacia are reemerging in Western societies, particularly in young children and in adolescents of African or Asian descent. Hypocalcemic seizures resulting from vitamin D deficiency are rare in adolescents, whereas fractures caused by seizures without evidence of direct trauma have not yet been reported in this population. We present an unusual case of secondary bilateral femoral fractures caused by hypocalcemic seizures in a 17 year-old boy with primary vitamin D deficiency. We examine the epidemiology and the clinical presentation of rickets and osteomalacia in the adolescent population, the risk of secondary injuries in patients with seizures, and the evaluation and management of hypocalcemic seizures and primary vitamin D deficiency. PMID- 17079598 TI - Prevention and management of pain in the neonate: an update. AB - The prevention of pain in neonates should be the goal of all caregivers, because repeated painful exposures have the potential for deleterious consequences. Neonates at greatest risk of neurodevelopmental impairment as a result of preterm birth (ie, the smallest and sickest) are also those most likely to be exposed to the greatest number of painful stimuli in the NICU. Although there are major gaps in our knowledge regarding the most effective way to prevent and relieve pain in neonates, proven and safe therapies are currently underused for routine minor yet painful procedures. Every health care facility caring for neonates should implement an effective pain-prevention program, which includes strategies for routinely assessing pain, minimizing the number of painful procedures performed, effectively using pharmacologic and nonpharmacologic therapies for the prevention of pain associated with routine minor procedures, and eliminating pain associated with surgery and other major procedures. PMID- 17079599 TI - Patient- and family-centered care and the role of the emergency physician providing care to a child in the emergency department. AB - Patient- and family-centered care is an approach to health care that recognizes the role of the family in providing medical care; encourages collaboration between the patient, family, and health care professionals; and honors individual and family strengths, cultures, traditions, and expertise. Although there are many opportunities for providing patient- and family-centered care in the emergency department, there are also challenges to doing so. The American Academy of Pediatrics and the American College of Emergency Physicians support promoting patient dignity, comfort, and autonomy; recognizing the patient and family as key decision-makers in the patient's medical care; recognizing the patient's experience and perspective in a culturally sensitive manner; acknowledging the interdependence of child and parent as well as the pediatric patient's evolving independence; encouraging family-member presence; providing information to the family during interventions; encouraging collaboration with other health care professionals; acknowledging the importance of the patient's medical home; and encouraging institutional policies for patient- and family-centered care. PMID- 17079600 TI - Menstruation in girls and adolescents: using the menstrual cycle as a vital sign. AB - Young patients and their parents often are unsure about what represents normal menstrual patterns, and clinicians also may be unsure about normal ranges for menstrual cycle length and amount and duration of flow through adolescence. It is important to be able to educate young patients and their parents regarding what to expect of a first period and about the range for normal cycle length of subsequent menses. It is equally important for clinicians to have an understanding of bleeding patterns in girls and adolescents, the ability to differentiate between normal and abnormal menstruation, and the skill to know how to evaluate young patients' conditions appropriately. Using the menstrual cycle as an additional vital sign adds a powerful tool to the assessment of normal development and the exclusion of pathological conditions. PMID- 17079601 TI - Much pain, little gain from voiding cystourethrograms after urinary tract infection. PMID- 17079602 TI - Effects of graduated driver licensing on fatalities in 16-year-olds. PMID- 17079603 TI - Pediatric disaster preparedness: what do we do next? PMID- 17079604 TI - Rotavirus gastroenteritis leading to secondary bacteremia in previously healthy infants. PMID- 17079605 TI - Azithromycin for pityriasis. PMID- 17079606 TI - Consumer-driven health plans: not all bad. PMID- 17079607 TI - The effects of marriage, civil union, and domestic partnership laws on the health and well-being of children. PMID- 17079608 TI - Prenatal exposure to conjugated bilirubin. PMID- 17079609 TI - Poverty, food insecurity, and obesity in children. PMID- 17079610 TI - Pediatric malpractice. PMID- 17079611 TI - The prevalence of dextromethorphan abuse among high school students. PMID- 17079612 TI - Implementation and case-study results of potentially better practices for family centered care: the family-centered care map. AB - OBJECTIVE: The objective of this study was to enhance the ability to coordinate and deliver care in a holistic manner, through a family-centered care map, so that the developmental, physical, and psychosocial needs of the infant and family are met. METHODS: A Web-based map was based on 7 distinct clinical phases with 3 variations of an infant's course through a NICU. Sixty-three potentially better practices were identified and 7 potentially better practices were implemented through case studies. RESULTS: Measures of family satisfaction revealed improvements in delivery of family-centered care. Increases in discharge growth parameters for extremely low birth weight infants were demonstrated. Length of stay for very low birth weight infants decreased from 73 to 60 days in Vermont. CONCLUSIONS: The collaborative process enhances identification of potentially better practices and results in both qualitative and quantitative improvements in family-centered care. PMID- 17079613 TI - Evaluation and development of potentially better practices to improve the discharge process in the neonatal intensive care unit. AB - OBJECTIVE: Our goal was to identify potentially better practices that create a successful discharge planning process that spans the entire newborn intensive care stay to the next level of care by embedding the discharge planning into all aspects of patient care and communication. METHODS: Potentially better practices were developed through recommendations from a content expert and a literature review. Internal benchmarking, self-assessment tools, monthly conference calls, the Neonatal Intensive Care Quality Improvement Collaborative 2002 listserv, parent feedback, and semiannual conferences were used to finalize recommendations and implement practices. RESULTS: Potentially better practices included (1) create an easy-to-use/easy-to-access discharge planning tool kit, (2) restructure written and oral communication tools and processes to reflect plans for the day, the stay, and the way to discharge, (3) maximize the impact and use of caregiver educational tools, and update materials and delivery systems for caregiver education, (4) use continuous quality improvement tools and processes to ensure parent/caregiver and staff satisfaction, and (5) analyze and enhance transfers into and interactions with the community. CONCLUSION: The potentially better practices are recommendations that are designed to integrate organizational, clinical, and operational processes to ensure optimal discharge planning from admission through follow-up in the community. PMID- 17079614 TI - Implementation and case-study results of potentially better practices to improve the discharge process in the neonatal intensive care unit. AB - OBJECTIVE: The objective of this study was to implement potentially better practices for discharge planning in the NICU. METHODS: Each participating hospital completed a self-assessment tool on discharge planning and a staff satisfaction survey. Parent satisfaction data were obtained from an Internet based survey. Many projects regarding discharge planning were completed at each participating center. A major emphasis was the development of transition points to span discharge planning over the entire hospitalization. Results of compliance with tasks or processes that were identified by the transition points and results of staff and parent satisfaction surveys were monitored over time. RESULTS: The implementation of the transition points at each center demonstrated an improvement in the completion of discharge tasks within the recommended time frame. Combined results of all centers demonstrated a moderate improvement in compliance with transition points from baseline to final measurement in the following areas: unit orientation (56%-81%), identification of a parent feeding plan (74%-92%), completion of cardiopulmonary resuscitation training (55%-72%), and car seat education (42%-63%). Staff survey results showed improvement from baseline to final measurement in the following areas: staff satisfaction with the discharge process (32%-50%), clear documentation of the discharge plan (26%-40%), and clarity of team members' roles in the discharge process (24%-44%). A resource kit on discharge planning was developed for staff and included a section with parent education material. An Internet-based parent satisfaction survey was implemented successfully. CONCLUSIONS: All centers that participated in the collaborative made significant strides in the discharge planning process. Overall, parent satisfaction with discharge planning was high, and improvements were noted in staff satisfaction and availability of resource material. PMID- 17079615 TI - Evaluation and development of potentially better practices for staffing in neonatal intensive care units. AB - OBJECTIVE: Five NICUs that participate in the Vermont Oxford Network's Neonatal Intensive Care Quality Improvement Collaborative 2002 attempted to identify potentially better practices that would have a directly impact on nurse recruitment and retention. The group identified nurse recruitment and retention as an important initiative for many hospitals that face a nursing shortage. METHODS: The group analyzed information from hospital demographics, literature reviews, process analysis questionnaires, and site visits. RESULTS: The literature review, process analysis questionnaire, and benchmarking with magnet hospitals identified 5 drivers for retention and recruitment. The drivers evolved into 5 potentially better practices that cover orientation, recognition and rewards, work environment, nurse/physician collaboration, and nursing autonomy. The magnet hospitals, which are known to have the highest retention rate and the lowest turnover rate, have many of these potentially better practices in place. CONCLUSION: The 5 practices described herein have the potential to decrease nursing turnover in NICUs. PMID- 17079616 TI - Implementation and case-study results of potentially better practices for staffing in neonatal intensive care units. AB - OBJECTIVE: Five NICUs that participate in the Vermont Oxford Network Quality Improvement Collaborative have implemented several potentially better practices in an attempt to decrease nurse turnover by 50%. These potentially better practices focus on orientation, rewards and recognition, healthy work environment, nurse-physician collaboration, and nursing autonomy. METHODS: Each unit implemented some or all of the potentially better practices. An Excel spreadsheet tool for tracking turnover rates was developed and used to measure the impact of the potentially better practices on retention. Rates were measured quarterly. RESULTS: After implementation of the potentially better practices, turnover rates fell at all of the NICUs ranging from 13% to 64%. CONCLUSIONS: Nurse retention is multifactorial. Implementation of the potentially better practices had a positive influence on nurse satisfaction but a varied impact on nurse retention. The impact of larger issues such as pay and staffing levels is significant and may not be influenced at the unit level. PMID- 17079618 TI - Implementation and case-study results of potentially better practices for collaboration between obstetrics and neonatology to achieve improved perinatal outcomes. AB - OBJECTIVE: The objective of this study was to make improvements in communication and collaboration between neonatal and obstetric specialties. Five NICUs from the Vermont Oxford Network's Evidence-Based Quality Improvement Collaborative in Neonatal and Perinatal Medicine tested potentially better practices that overlap obstetric and NICU care. METHODS: One area of practice improvement was the management of the pregnancy at the margin of viability. Another included the use of team training and video simulation to improve team performance during high risk deliveries using aviation-based communication techniques. Another focus of the collaborative was the creation of a multicenter database to measure combined perinatal and neonatal outcomes. RESULTS: The principle outcomes are increased patient satisfaction with teamwork between neonatology and obstetric services and improved team response times for emergent deliveries and the increased use of team communication skills during video simulations of high-risk deliveries. CONCLUSIONS: Implementing these potentially better practices can result in improved communication and collaboration related to perinatal and neonatal care. PMID- 17079617 TI - Evaluation and development of potentially better practices for perinatal and neonatal communication and collaboration. AB - OBJECTIVE: The obstetric and neonatal exploratory focus group of the Vermont Oxford Network Neonatal Intensive Care Quality Improvement Collaborative 2002 set out to improve collaboration, communication, and coordination between maternal and neonatal caregivers in 3 areas: the pregnancy at 22 to 26 weeks, measurement of maternal outcomes that are linked with neonatal outcomes, and team performance during high-risk delivery. Antepartum and intrapartum maternal attributes and interventions also were considered important measurements to identify practice variations and their relationship to neonatal outcomes for ongoing obstetric and neonatal collaboration. METHODS: Potentially better practices were developed on the basis of evidence in the literature, expert opinion, and internal analysis at the participating perinatal centers. The potentially better practices include development of local guidelines at each center for the care and counseling of pregnant women who are at risk for delivering at the margin of viability; communication strategies for obstetric and neonatology providers relating to high risk pregnancy treatment plans; team communication and performance at high-risk deliveries; design of organizational structures and processes that facilitate obstetric and neonatal collaboration; and development of perinatal data to evaluate effects of perinatal practices on maternal, fetal, and neonatal outcomes. RESULTS: As a result of the project, participating centers developed local guidelines for pregnancies between 22 and 26 weeks, created a cross-center maternal database that currently is being linked to neonatal outcomes, and completed a pilot study on video simulation of neonatal-perinatal team communication. CONCLUSIONS: Increased understanding of practice variation in the management of care for infants who are at the margins of viability, locally developed guidelines, and a focus on improved team communication during delivery can be accomplished with a multicenter collaborative approach. PMID- 17079619 TI - Management of high-order multiple births: application of lessons learned because of participation in Vermont Oxford Network collaboratives. AB - OBJECTIVES: The delivery and care of sextuplets is complex. Potentially better practices that were developed as part of the Vermont Oxford Network improvement collaboratives were used to prepare for a sextuplet delivery at Akron Children's Hospital. METHODS: The team used potentially better practices that were learned from the Neonatal Intensive Care Quality Improvement Collaborative 2002 using multidisciplinary teams. There was extensive media coverage of the delivery. RESULTS: The goal was to use nearly all potentially better practices that focused on the goals of reducing nosocomial infection, reducing chronic lung disease, reducing radiograph use, reducing length of stay, reducing blood gas use, promoting nutrition, reducing intraventricular hemorrhage, and enriching family centered care. The center aimed to use these 97 potentially better practices. Of the 97 possible potential better practices as set by the Neonatal Intensive Care Quality Improvement Collaborative 2002, 96 (99%) were used. CONCLUSIONS: This is a blueprint that any center that is faced with high-order multiple births could use as a reference point to begin planning. The team created a benchmark to achieve in every birth of very low birth weight infants and not just a special situation of high-order multiple births. PMID- 17079620 TI - Attitudes of obstetric and pediatric health care providers toward resuscitation of infants who are born at the margins of viability. AB - OBJECTIVES: The objective of this study was to determine the attitudes of a variety of health care providers toward the recommendations that should be made to parents regarding the resuscitation of infants who are born at the margins of viability. METHODS: A written questionnaire was distributed to the medical and nursing staff at 4 tertiary perinatal centers. For each of 5 weekly gestational age intervals from 22 weeks to 26 weeks, 6 days, the health care providers were asked to describe on a scale from 1 to 5 whether they would strongly discourage through strongly encourage resuscitation. They also were queried regarding their comfort with counseling regarding these issues. The attitudes of various groups of providers were compared across weekly intervals. RESULTS: A total of 204 physicians and 539 nurses completed the survey. The majority would strongly discourage, either discourage or strongly discourage, be neutral or recommend, recommend or strongly recommend, and strongly recommend resuscitation during the 23rd, 24th, 25th, 26th, and 27th weeks of gestation, respectively. Obstetric caregivers were slightly less likely than pediatric caregivers to strongly discourage resuscitation from 22 weeks to 22 weeks, 6 days and 23 weeks to 23 weeks, 6 days. There were no significant differences in the recommendations of obstetricians and pediatricians. Pediatric nurses were more likely to strongly recommend resuscitation from 26 weeks to 26 weeks, 6 days and more likely either to discourage or to strongly discourage resuscitation from 23 weeks to 23 weeks, 6 days and to strongly discourage resuscitation from the 22 weeks to 22 weeks, 6 days than their obstetric counterparts. Obstetric nurses were slightly less likely than obstetricians to strongly recommend resuscitation at 26 weeks to 26 weeks, 6 days and less likely to strongly discourage resuscitation from 22 weeks to 22 weeks, 6 days. CONCLUSIONS: The caregivers' recommendations seem to be based logically on the current literature regarding survival and morbidity that is experienced by infants who are born at the threshold of viability. Although there are minor differences, there was a relatively consistent approach among professional groups. PMID- 17079621 TI - Overcoming barriers to oxygen saturation targeting. AB - OBJECTIVE: To reduce hyperoxia in very low birth weight infants who receive supplemental oxygen, the Children's Mercy Hospital neonatal respiratory quality improvement committee introduced the potentially better practice of oxygen saturation targeting and identified strategies to overcome barriers to implementation of this practice. METHODS: Using rapid-cycle quality improvement projects, this center adapted an oxygen saturation targeting protocol and tracked hourly oxygen saturation as measured by pulse oximetry in very low birth weight infants who received supplemental oxygen. RESULTS: The percentage of time in the range of 90% to 94% of oxygen saturation as measured by pulse oximetry increased from 20% to an average of 35% after implementation of the protocol. The percentage of time with oxygen saturation as measured by pulse oximetry >98% dropped from 30% to an average of 5% to 10%. CONCLUSIONS: A well-planned strategy for implementing oxygen saturation targeting can result in a sustained change in clinical practice as well as change in the culture of the NICU regarding the use of oxygen. PMID- 17079622 TI - Standardizing nasal cannula oxygen administration in the neonatal intensive care unit. AB - OBJECTIVE: A multicycle, quality improvement method was used to standardize nasal cannula O2 administration and weaning in the NICU. METHODS: A 2-armed nasal cannula standardized order form (nasal cannula for stable O2 arm and nasal cannula for stable flow arm) was developed after review of the literature, surveying of the practice of NICU physicians and nurse practitioners, and development of consensus among these providers. Outcomes were measured by tracking the distribution of protocol arm chosen, days on O2, weeks on nasal cannula, and disposition of infants who were supported by nasal cannula. Data were collected in an SPSS statistical data set. RESULTS: Of the 90 infants evaluated, 12 were supported on the stable O2 arm and 53 on the stable flow arm for their entire nasal cannula course. Twenty-five infants switched between arms of support. Patients who were on the stable flow arm of the standard order set for their entire nasal cannula course experienced fewer O2 days but more days on nasal cannula. A subpopulation of infants were supported on nasal cannula flow 0.5 to 1.0 L, with fraction of inspired O2 of 21%. When data from the first 10 weeks of observation were compared with that of the second 10 weeks, the rate of discharge on O2 had decreased from 13 (30%) of 44 to 3 (7%) of 39. CONCLUSIONS: The multiple steps of literature review, practice surveys, and consensus-building resulted in enthusiastic reception of the nasal cannula standardized order form. The 2-armed nasal cannula protocol forced caregivers to consider which method of support was most beneficial for each infant who was on nasal cannula and allowed a subpopulation of NICU patients to be supported with a lower fraction of inspired O2 than previously used in the NICU. PMID- 17079623 TI - Sucrose analgesia: identifying potentially better practices. AB - OBJECTIVE: The objectives of this study were to review the use of oral sucrose for procedural pain management in NICUs, develop potentially better practice guidelines that are based on the best current evidence, and provide ideas for the implementation of these potentially better practices. METHODS: A collaboration of 12 centers of the Vermont Oxford Network worked together to review the strength of the evidence, clinical indications, dosage, administration, and contraindications and identify potential adverse effects for the use of sucrose analgesia as the basis of potentially better practices for sucrose analgesia guidelines. Several units implemented the guidelines. RESULTS: Through reviews and inputs from all centers of the evidence, consensus was reached and guidelines that included indication, dosage per painful procedure, age-related dosage over 24 hours, method of delivery, and contraindications were developed. CONCLUSIONS: Guidelines now are available from a consensus group, and suggestions for implementation of guidelines, based on implementation of other pain management strategies, were developed. PMID- 17079625 TI - Evaluation and development of potentially better practices to reduce bronchopulmonary dysplasia in very low birth weight infants. AB - OBJECTIVE: The objective of this study was to describe development and implementation of potentially better practices to reduce bronchopulmonary dysplasia in very low birth weight infants (birth weight: 501-1500 g). METHODS: Results of Breathsavers Group meetings, conference calls and critically appraised topic summaries were used to construct potentially better practices. Implementation plans and experiences were reported by participants and collated. RESULTS: The Breathsavers Group developed 13 potentially better practices, based on published evidence and expert opinion. Participants determined which potentially better practices to implement and implementation methods. Participating NICUs implemented an average of 5 potentially better practices (range: 3-9). The Breathsavers Group also developed a resource kit, identified common obstacles to implementation, and initiated research to define bronchopulmonary dysplasia better. CONCLUSIONS: Multiinstitutional collaboration facilitated development and implementation of potentially better practices to reduce bronchopulmonary dysplasia. PMID- 17079626 TI - Reduction of bronchopulmonary dysplasia after participation in the Breathsavers Group of the Vermont Oxford Network Neonatal Intensive Care Quality Improvement Collaborative. AB - OBJECTIVE: The objective of this study was to compare the primary and secondary outcomes of very low birth weight infants before and after participation in the Breathsavers Group of the Vermont Oxford Network-sponsored Neonatal Intensive Care Quality Collaborative. METHODS: Hospitals that participated in the Breathsavers Group contributed clinical data on the outcomes of their very low birth weight infants to the Vermont Oxford Network using standardized clinical definitions, data forms, and inclusion criteria. Outcomes from the last year of the collaborative, 2003, were compared with those from the baseline year, 2001. Models for treatment practices and outcomes measures were adjusted for within hospital correlation (clustering) and standard risk factors that were present at birth. RESULTS: Bronchopulmonary dysplasia dropped significantly in 2003 compared with the baseline year. Survival improved but not significantly. In addition, severe retinopathy of prematurity, severe intraventricular hemorrhage, and supplemental oxygen at discharge dropped significantly. The use of conventional ventilation at any time during the initial hospitalization, postnatal steroids, and time to first dose of surfactant all decreased significantly. The use of nasal continuous positive airway pressure at any time during hospitalization increased. The use of high-frequency ventilation, delivery room intubation, and surfactant at any time during hospitalization did not change. CONCLUSIONS: The Breathsavers Group improved both clinical care processes and clinical outcomes during the Neonatal Intensive Care Quality Collaborative. PMID- 17079627 TI - Evaluation and development of potentially better practices to improve pain management of neonates. AB - OBJECTIVE: Despite increased knowledge, improved options, and regulatory mandates, pain management of neonates remains inadequate, promoted by the ineffective translation of research data into clinical practice. The Neonatal Intensive Care Quality Improvement Collaborative 2002 was created to provide participating NICUs the tools necessary to translate research, related to prevention and treatment of neonatal pain, into practice. The objective for this study was to use proven quality improvement methods to develop a process to improve neonatal pain management collaboratively. METHODS: Twelve members of the Neonatal Intensive Care Quality Improvement Collaborative 2002 formed an exploratory group to improve neonatal pain management. The exploratory group established group and site-specific goals and outcome measures for this project. Group members crafted a list of potentially better practices on the basis of the available literature, encouraged implementation of the potentially better practices at individual sites, developed a database for sharing information, and measured baseline outcomes. RESULTS: The goal "improve the assessment and management of infants experiencing pain in the NICU" was established. In addition, each site within the group identified local goals for improvement in neonatal pain management. Data from 7 categories of neonates (N = 277) were collected within 48 hours of NICU admission to establish baseline data for clinical practices. Ten potentially better practices were developed for prioritized pain conditions, and 61 potentially better practices were newly implemented at the 12 participating sites. Various methods were used for pain assessment at the participating centers. At baseline, heel sticks were used more frequently than peripheral intravenous insertions or venipunctures, with substantial variability in the number of avoidable procedures between centers. Pain was assessed in only 17% of procedures, and analgesic interventions were performed in 19% of the procedures at baseline. CONCLUSIONS: Collaborative use of quality improvement methods resulted in the creation of self-directed, efficient, and effective processes to improve neonatal pain management. Group establishment of potentially better practices, collective and site-specific goals, and extensive baseline data resulted in accelerated implementation of clinical practices that would not likely occur outside a collaborative setting. PMID- 17079628 TI - Implementation and case-study results of potentially better practices to improve pain management of neonates. AB - OBJECTIVE: Collaborative quality improvement techniques were used to facilitate local quality improvement in the management of pain in infants. Several case studies are presented to highlight this process. METHODS: Twelve NICUs in the Neonatal Intensive Care Quality Improvement Collaborative 2002 focused on improving neonatal pain management and sedation practices. These centers developed and implemented evidence-based potentially better practices for pain management and sedation in neonates. The group introduced changes through plan-do study-act cycles and tracked performance measures throughout the process. RESULTS: Strategies for implementing potentially better practices varied between centers on the basis of local characteristics. Individual centers identified barriers to implementation, developed tools for improvement, and shared their experience with the collaborative. Baseline data from the 12 sites revealed substantial opportunities for improved pain management, and local potentially better practice implementation resulted in measurable improvements in pain management at participating centers. CONCLUSIONS: The use of collaborative quality improvement techniques enhanced local quality improvement efforts and resulted in effective implementation of potentially better practices at participating centers. PMID- 17079629 TI - Development and dissemination of potentially better practices for the provision of family-centered care in neonatology: the family-centered care map. AB - OBJECTIVE: Family-centered care has become integral to the provision of quality neonatal intensive care. However, practices that reflect the core principles of family-centered care have not been described fully in the literature or implemented and evaluated consistently within newborn intensive care. The objective of this study was to create a family-centered care map that enhances the ability of the health care team to work with families to coordinate and deliver care in a holistic manner to meet the developmental, physical, and psychosocial needs of NICU patients and their families. METHODS: Potentially better practices were developed for sequential clinical phases by using standardized methods. These included focus groups with families, brainstorming sessions with staff, literature review, and input from established family advisory groups and family-centered care experts. Potentially better practices then were integrated into the family-centered care map that was configured in a Web-based format. Overall utility will be evaluated by determining the effect of the family-centered care map on length of stay, parental satisfaction, and family centered care beliefs and practices among NICU staff. RESULTS: Sixty-three potentially better practices were identified for 7 clinical phases and 3 variations that were believed to characterize the clinical course of a typical NICU patient. A prototype of the Web-based family-centered care map that illustrates the clinical phases with links to the related potentially better practices, operational processes, and case studies was created. Baseline data from a care provider survey, from a family satisfaction survey, and on length of stay have been collected. CONCLUSIONS: Quality improvement methods and collaboration among 3 centers led to the development of an innovative Web-based resource to assist individual care providers and family advisors to provide comprehensive family-centered care to infants and families. Implementation of the family-centered care map has potential to affect positively the quality of newborn intensive care and lead to improved long-term outcomes. PMID- 17079631 TI - Is propofol neurotoxic to the developing brain? PMID- 17079632 TI - Drug transformation: Advances in drug delivery systems. PMID- 17079633 TI - Propofol induces growth cone collapse and neurite retractions in chick explant culture. AB - PURPOSE: Propofol neurotoxicity has been demonstrated in several cell culture systems. This study was undertaken to determine whether propofol has neurotoxic effects on peripheral, retinal, and autonomic neurons, and which neurons are particularly liable to injury by propofol. METHOD: Dorsal root ganglia, retinal ganglion cell layers, and sympathetic ganglion chains were isolated from day eight chick embryos and cultured for 20 hr. Thereafter, propofol was added at various concentrations [5-300 microM (0.9-53 microg x mL(-1))] to investigate its effects on these three types of neuronal tissue. Morphological changes were examined quantitatively by growth cone collapse assay. Propofol concentrations were measured using high performance liquid chromatography. RESULTS: Propofol induced growth cone collapse and neurite destruction. The three types of neurons tested exhibited significantly different dose-response relationships two hours after the application of propofol (P < 0.001) but not at 24 hr after application. The growth cone-collapsing effect was at least partially reversible in all three types of neurons after exposure to 100 microM propofol up to six hours, though reversibility was not observed after 24-hr exposure. CONCLUSION: While the clinical safety profile of propofol has been well documented, at high concentrations propofol has potential neurotoxicity on growing neurons in vitro. PMID- 17079634 TI - Documentation on the anesthetic record: Correlation with clinically important variables. AB - PURPOSE: A survey was undertaken at a single Academic Health Sciences Centre to document the opinions of anesthesiologists regarding what variables are important to document on the anesthetic record. A subsequent chart review of anesthetic records was undertaken to determine the extent to which these anesthesiologists record the variables they consider important. METHODS: A survey form was mailed to all practicing staff anesthesiologists at the four adult McGill University affiliated hospitals. Anesthesiologists were asked to rank 23 preoperative and 33 intraoperative variables on a scale from 1-5: (1 = essential; 2 = important; 3 = useful; 4 = not important; 5 = excessive information). All variables considered by consensus < or = 2 (important to essential) were then assessed as to whether they were recorded on 60 charts randomly selected from each of the four teaching hospitals. Only anesthetic records completed by staff anesthesiologists were evaluated. RESULTS: Ninety percent (47/52) of survey forms were completed and returned. Preoperative variables considered most important to document included examination of the patient's airway and allergy status. Intraoperative variables considered most important for documentation were the patient's vital signs. The only variable to have been recorded on all the anesthetic records was the anesthesiologist's name. The allergy status was the most recorded preoperative variable (84% of charts). The recording rates of intraoperative variables ranged from 100% (anesthesiologist's name, start time of anesthesia) to 24% (estimated blood loss). CONCLUSION: McGill anesthesiologists consider many preoperative and intraoperative variables to be important to document on the anesthetic record. However, subsequent chart review indicated that many of these variables are recorded inconsistently. The transmission of anesthesia-related medical information might be improved if anesthesiologists recorded more consistently information they consider to be important. PMID- 17079635 TI - Liposomal formulations of prilocaine, lidocaine and mepivacaine prolong analgesic duration. AB - PURPOSE: A laboratory investigation was undertaken to compare the in vivo antinociceptive effects of 2% liposomal formulations of prilocaine (PLC), lidocaine (LDC) and mepivacaine (MVC) compared to plain solutions of each of these three local anesthetics. METHODS: Large unilamellar vesicles were prepared by extrusion (400 nm), at pH 7.4. The membrane/water partition coefficients were obtained from encapsulation efficiency values, after incorporation of each local anesthetic to the vesicles. The anesthetic effect of each liposomal formulation was compared to the respective local anesthetic solution in water, using the infraorbital nerve-blockade test, in rats. RESULTS: The partition coefficients were: 57 for PLC, 114 for LDC and 93 for MVC. In vivo results showed that local anesthetic-free liposomes, used as control, had no analgesic effect. In contrast, the encapsulated formulations induced increased intensities of total anesthetic effect (35.3%, 26.1% and 57.1%) and time for recovery (percentage increases of 30%, 23.1% and 56%), respectively, for PLC, LDC and MVC when compared to the plain solutions (P < 0.01). CONCLUSIONS: These results indicate that liposomes provide effective drug-delivery systems for intermediate-duration local anesthetics. Mepivacaine was affected to the greatest extent, while LDC benefited least from liposome encapsulation, possibly due to greater vasodilatory properties of LDC. PMID- 17079636 TI - Compressed air injection technique to standardize block injection pressures. AB - PURPOSE: Presently, no standardized technique exists to monitor injection pressures during peripheral nerve blocks. Our objective was to determine if a compressed air injection technique, using an in vitro model based on Boyle's law and typical regional anesthesia equipment, could consistently maintain injection pressures below a 1293 mmHg level associated with clinically significant nerve injury. METHODS: Injection pressures for 20 and 30 mL syringes with various needle sizes (18G, 20G, 21G, 22G, and 24G) were measured in a closed system. A set volume of air was aspirated into a saline-filled syringe and then compressed and maintained at various percentages while pressure was measured. The needle was inserted into the injection port of a pressure sensor, which had attached extension tubing with an injection plug clamped "off". Using linear regression with all data points, the pressure value and 99% confidence interval (CI) at 50% air compression was estimated. RESULTS: The linearity of Boyle's law was demonstrated with a high correlation, r = 0.99, and a slope of 0.984 (99% CI: 0.967-1.001). The net pressure generated at 50% compression was estimated as 744.8 mmHg, with the 99% CI between 729.6 and 760.0 mmHg. The various syringe/needle combinations had similar results. CONCLUSION: By creating and maintaining syringe air compression at 50% or less, injection pressures will be substantially below the 1293 mmHg threshold considered to be an associated risk factor for clinically significant nerve injury. This technique may allow simple, real-time and objective monitoring during local anesthetic injections while inherently reducing injection speed. PMID- 17079637 TI - Intravenous nalbuphine 50 microg x kg(-1) is ineffective for opioid-induced pruritus in pediatrics. AB - PURPOSE: This investigation evaluated the efficacy of nalbuphine in treating postoperative opioid-induced pruritus (Pr) in pediatric patients. METHODS: After Ethics Board approval, the dual site, tertiary care teaching centre study recruited 212 subjects, age > or = seven years, who received opioid analgesia postoperatively. A modified, self-report colour analogue scale (CAS) scored pruritus intensity (PrI). Subjects who reported PrI score > or = 5/10 were randomized to treatment with nalbuphine 50 microg x kg(-1) iv (max 5 mg) or saline placebo. A pruritus intensity difference (PrID) > or = 50% was considered a positive outcome. RESULTS: Of 260 subjects approached, 212 consented and 184 received opioids. Median age was 13 yr (range 7-19) and median weight was 51 kg (range 19.6-134.8 kg). Pruritus intensity > or = 5/10 occurred in 37 (20.1%) subjects. Intravenous morphine [patient-controlled analgesia (PCA)/continuous infusion] was associated with Pr in 68% of subjects over a wide dose range (9.4 63.2 mug.kg(-1).hr(-1)). Pruritus occurred in 36% of patients in the PCA group compared to continuous opioid infusion (27%) and epidural administration (27%). Pruritus intensity difference > or = 50% was achieved in 55.6% of nalbuphine and 57.9% of saline-treated subjects. CONCLUSION: This preliminary report suggests that nalbuphine 50 microg x kg(-1) iv is not effective in treating postoperative opioid-induced pruritus in pediatric patients. The modified CAS score and PrID warrant further investigation. PMID- 17079638 TI - Topical lidocaine and oral acetaminophen provide similar analgesia for myringotomy and tube placement in children. AB - PURPOSE: Preoperative oral acetaminophen (30 mg x kg(-1)) was compared with topical 2% lidocaine ear drops for postoperative analgesia following bilateral myringotomy and tube placement (BMT) in children. METHODS: In a randomized, prospective, double-blind trial, we studied 124 patients, six months to eight years, ASA physical status I or II, undergoing elective BMT under general anesthesia. The patients in Group I received acetaminophen 30 mg x kg(-1) orally in a grape flavoured syrup 30 to 60 min before surgery and 0.9% saline drops (placebo) in each ear upon insertion of tympanostomy tube. Patients in Group II received a placebo (grape flavoured syrup) before surgery and 2% lidocaine, 0.5 mL in each ear when ear tubes were inserted. Postoperative pain assessments were recorded every five minutes in the postanesthesia care unit, and every 15 min in the day care surgical unit (DCSU) using the modified Children's Hospital of Eastern Ontario pain scale (mCHEOPS), a ten-point scale. Pain at home was documented by parents using a 0 (no pain) to 10 (worst pain imaginable) scale. RESULTS: The median (range) mCHEOPS scores in the DCSU at 15 and 30 min were similar, i.e., 5 (4-9) in the acetaminophen group and 4 (4-8) in the lidocaine group. The proportion of patients receiving supplemental analgesics in the 24 hr following surgery was similar in both groups (45% and 42% respectively). CONCLUSION: Topical lidocaine and oral acetaminophen in a dose of 30 mg x kg(-1) provide similar analgesia following BMT. PMID- 17079639 TI - Images in Anesthesia: Ex utero intrapartum treatment (exit procedure): fetal airway management. PMID- 17079641 TI - Review article: The role of statins in reducing perioperative cardiac risk: Physiologic and clinical perspectives. AB - PURPOSE: To review the pathobiology and clinical implications of coronary vulnerable atherosclerotic plaques (VAPs), to discuss the role of statin therapy in VAP stabilization, and the potential benefits of perioperative statin therapy (PST) in reducing perioperative risk of acute coronary syndromes (ACSs). SOURCE: MEDLINE search using "perioperative", "cardiac morbidity", "atherosclerosis", "vulnerable plaque", "statins" and combinations of these terms as keywords. The reference lists of relevant articles were further reviewed to identify additional citations. PRINCIPAL FINDINGS: The nonstenotic, yet rupture-prone VAP causes most myocardial infarctions (MIs) and other ACSs, both in the nonsurgical and surgical patients. Large clinical trials in both primary and secondary prevention and in patients with ACSs have demonstrated that statin therapy will reduce cardiovascular morbidity and mortality across a broad spectrum of patient subgroups. These trials also suggest, and laboratory investigations establish, that statins possess favourable vascular effects independent of cholesterol reduction. Statins appear to interfere specifically with the pathophysiologic mechanisms implicated in atherothrombotic disease. Statins reduce vascular inflammation, improve endothelial function, stabilize VAPs, and reduce platelet aggregability and thrombus formation. Recent studies have shown that PST is associated with a reduced incidence of perioperative and long-term cardiovascular complications in high-risk patients. Combined therapy with statins and ss blockers is a conceptually valid strategy targeting critical steps in the pathogenesis of an ACS. CONCLUSION: Emerging evidence for the efficacy and safety of PST is promising, especially when combined with ss-blocker therapy in patients at highest risk. Confirmation of this early evidence awaits the results of ongoing and future prospective randomized controlled trials. PMID- 17079640 TI - Effects of halothane, sevoflurane and desflurane on the force-frequency relation in the dog heart in vivo. AB - PURPOSE: Frequency potentiation is the increase in force of contraction induced by an increased heart rate (HR). This positive staircase phenomenon has been attributed to changes in Ca2+ entry and loading of intracellular Ca2+ stores. Volatile anesthetics interfere with Ca2+ homeostasis of cardiomyocytes. We hypothesized that frequency potentiation is altered by volatile anesthetics and investigated the influence of halothane (H), sevoflurane (S) and desflurane (D) on the positive staircase phenomenon in dogs in vivo. METHODS: Dogs were chronically instrumented for measurement of left ventricular (LV) pressure and cardiac output. Heart rate was increased by atrial pacing from 120 to 220 beats x min(-1) and the LV maximal rate of pressure increase (dP/dt(max)) was determined as an index of myocardial performance. Measurements were performed in conscious dogs and during anesthesia with 1.0 minimal alveolar concentrations of each of the three inhaled anesthetics. RESULTS: Increasing HR from 120 to 220 beats x min(-1) increased dP/dt(max) from 3394 +/- 786 (mean +/- SD) to 3798 +/- 810 mmHg sec(-1) in conscious dogs. All anesthetics reduced dP/dt(max) during baseline (at 120 beats x min(-1): H, 1745 +/- 340 mmHg x sec(-1); S, 1882 +/- 418; D, 1928 +/- 454, all P < 0.05 vs awake) but did not influence the frequency potentiation of dP/dt(max) (at 220 beats x min(-1): H, 1981 +/- 587 mmHg x sec(-1); S, 2187 +/- 787; D, 2307 +/- 691). The slope of the regression line correlating dP/dt(max) and HR was not different between awake and anesthetized dogs. Increasing HR did not influence cardiac output in awake or anesthetized dogs. CONCLUSION: These results indicate that volatile anesthetics do not alter the force-frequency relation in dogs in vivo. PMID- 17079642 TI - Brief review: Nondepolarizing neuromuscular blocking drugs and critical illness myopathy. AB - PURPOSE: Critically-ill patients who receive nondepolarizing neuromuscular blocking drugs (NMBDs) may be at risk of developing profound muscle weakness that may last for months after the NMBD is discontinued, especially when large cumulative doses of NMBDs and corticosteroids are co-administered to septic, mechanically ventilated patients. This review focuses on the etiology and clinical features of critical illness myopathy (CIM), summarizes specific risk factors for its development, and discusses strategies that might be used to attenuate or even prevent the development of this potentially devastating syndrome. CLINICAL FEATURES: The etiology of CIM is unknown. Whether it can develop in at-risk patients who undergo lengthy operations during which they receive NMBDs is also unknown. In some patients following exposure to NMBDs their motor systems are impaired secondary to loss of thick (myosin) filaments that render the muscle unexcitable to direct electrical stimulation, while the sensory system is spared. Management of patients who develop NMBD myopathy is supportive, consisting of nutritional support, physical therapy, and daily trials of decreased ventilatory support. CONCLUSION: Recent guidelines recommend that NMBDs be used in critically ill patients only when absolutely necessary, that the depth of muscle paralysis be monitored to avoid overdosing and metabolite accumulation, and that drug administration be curtailed periodically to allow interruption of sustained NMBD effect. PMID- 17079643 TI - Best evidence in critical care medicine: Early antibiotics and survival from septic shock: it's about time. PMID- 17079644 TI - [Respiratory depression after intrathecal injection of tramadol with hyperbaric bupivacaine]. PMID- 17079645 TI - Styletted orotracheal intubation in difficult airway management. PMID- 17079646 TI - GlideScope videolaryngoscope facilitates nasotracheal intubation. PMID- 17079647 TI - Mandibular dislocation from yawning during induction of anesthesia. PMID- 17079648 TI - [A medial approach for a continuous sciatic block in the popliteal fossa]. PMID- 17079649 TI - Monocytes infected with Mycobacterium tuberculosis regulate MAP kinase-dependent astrocyte MMP-9 secretion. AB - Tuberculosis (TB) of the CNS (CNS-TB) carries a high mortality. Disease pathology is characterized by widespread destruction of CNS tissues. Matrix metalloproteinase-9 (MMP-9) is able to catabolyze specific components of the CNS tissue matrix and blood-brain barrier. Increased cerebrospinal fluid MMP-9 concentrations are associated with tissue damage, leukocyte infiltration, and death in CNS-TB. Using zymography, Western analysis, and transcription factor assays, we investigated mechanisms regulating MMP-9 activity in CNS-TB. We demonstrate that conditioned media from monocytes infected with Mycobacterium tuberculosis (CoMTB) induce MMP-9 secretion from astrocytes (U373-MG). IL-1beta and TNF-alpha are necessary but not sufficient for such induction of astrocyte MMP-9 secretion. CoMTB up-regulates AP-1 DNA-binding activity, and the c-Jun, FosB, and JunB subunits are particularly increased. MMP-9 secretion from CoMTB stimulated astrocytes is dependent on the activity of p38, Erk, and Jnk MAPKs. Phosphorylation of p38, Erk, and Jnk is activated rapidly, peaking 30 min poststimulation with CoMTB. Inhibition of IL-1beta but not TNF-alpha in CoMTB decreases p38, Erk, and Jnk activity in astrocytes. Consistently, IL-1beta signals through the MAPK cascade at physiological levels, whereas TNF-alpha, IL 6, IL-10, CCL-2, CCL-5, and CXCL-8 (all present in CoMTB) do not. In summary, the data suggest that monocyte-dependent cytokine networks may play a key role in the development of a matrix-degrading environment during CNS-TB. PMID- 17079650 TI - Thimerosal induces TH2 responses via influencing cytokine secretion by human dendritic cells. AB - Thimerosal is an organic mercury compound that is used as a preservative in vaccines and pharmaceutical products. Recent studies have shown a TH2-skewing effect of mercury, although the underlying mechanisms have not been identified. In this study, we investigated whether thimerosal can exercise a TH2-promoting effect through modulation of functions of dendritic cells (DC). Thimerosal, in a concentration-dependent manner, inhibited the secretion of LPS-induced proinflammatory cytokines TNF-alpha, IL-6, and IL-12p70 from human monocyte derived DC. However, the secretion of IL-10 from DC was not affected. These thimerosal-exposed DC induced increased TH2 (IL-5 and IL-13) and decreased TH1 (IFN-gamma) cytokine secretion from the T cells in the absence of additional thimerosal added to the coculture. Thimerosal exposure of DC led to the depletion of intracellular glutathione (GSH), and addition of exogenous GSH to DC abolished the TH2-promoting effect of thimerosal-treated DC, restoring secretion of TNF alpha, IL-6, and IL-12p70 by DC and IFN-gamma secretion by T cells. These data suggest that modulation of TH2 responses by mercury and thimerosal, in particular, is through depletion of GSH in DC. PMID- 17079651 TI - The reduced GM-CSF priming of ROS production in granulocytes from patients with myelodysplasia is associated with an impaired lipid raft formation. AB - Patients with myelodysplasia (MDS) show an impaired reactive oxygen species (ROS) production in response to fMLP stimulation of GM-CSF-primed neutrophils. In this study, we investigated the involvement of lipid rafts in this process and showed that treatment of neutrophils with the lipid raft-disrupting agent methyl-beta cyclodextrin abrogates fMLP-induced ROS production and activation of ERK1/2 and protein kinase B/Akt, two signal transduction pathways involved in ROS production in unprimed and GM-CSF-primed neutrophils. We subsequently showed that there was a decreased presence of Lyn, gp91(phox), and p22(phox) in lipid raft fractions from neutrophils of MDS. Furthermore, the plasma membrane expression of the lipid raft marker GM1, which increases upon stimulation of GM-CSF-primed cells with fMLP, was reduced significantly in MDS patients. By electron microscopy, we showed that the fMLP-induced increase in GM1 expression in GM-CSF-primed cells was a result of de novo synthesis, which was less efficient in MDS neutrophils. Taken together, these data indicate an involvement of lipid rafts in activation of signal transduction pathways leading to ROS production and show that in MDS neutrophils, an impaired lipid raft formation in GM-CSF-primed cells results in an impaired ROS production. PMID- 17079652 TI - Three-dimensional reconstruction and quantitative study of a pyramidal cell of a Cajal histological preparation. AB - The year 2006 marks the centenary of the Nobel Prize for Physiology or Medicine awarded to Santiago Ramon y Cajal and Camilo Golgi. We commemorate this centenary with a three-dimensional reconstruction and a quantitative study of a pyramidal cell of a Cajal's histological preparation. This preparation is one of the 4529 histological preparations personally made by Ramon y Cajal and preserved in the Museum Cajal. The three-dimensional reconstruction of the neuron allows visualizing one important discovery of Ramon y Cajal that constitutes an active field of research in present-day neuroscience: dendritic spines. PMID- 17079653 TI - Activity-dependent plasticity in the olfactory intrabulbar map. AB - In mammals, each olfactory bulb contains two mirror-symmetric glomerular maps. Isofunctional glomeruli within each bulb are specifically linked through a set of reciprocal intrabulbar projections (IBPs) to form an intrabulbar map. We injected neural tracers into the glomerular layer on one side of the bulb and examined the resulting projection on the opposite side. In adult mice, the size of the projection tuft is directly proportional to the size of the injected region. Using this ratio as a measure of IBP maturity, we find an immature 5:1 projection to injection ratio at 1 week of age that gradually refines to a mature 1:1 by 7 weeks. Moreover, whereas the glomerular map is able to form despite the elimination of odorant-induced activity, the intrabulbar map shows clear activity dependence for its precise formation. Here we show through experiments with both naris-occluded and anosmic mice that odorant-induced activity is not required to establish IBPs but is crucial for projection refinement. In contrast, increased glomerular activation through exposure to distinct odorants during map development can accelerate the refinement of projections associated with the activated glomeruli. These findings illustrate a clear role for odorant-induced activity in shaping the internal circuitry of the bulb. Interestingly, activity deprivation can alter the organization of both the developing and the mature map to the same degree, demonstrating that intrabulbar map plasticity is maintained into adulthood with no discernible critical period. PMID- 17079654 TI - Non-NMDA-type glutamate receptors are essential for maturation but not for initial assembly of synapses at Drosophila neuromuscular junctions. AB - The assembly of glutamatergic postsynaptic densities (PSDs) seems to involve the gradual recruitment of molecular components from diffuse cellular pools. Whether the glutamate receptors themselves are needed to instruct the structural and molecular assembly of the PSD has hardly been addressed. Here, we engineered Drosophila neuromuscular junctions (NMJs) to express none or only drastically reduced amounts of their postsynaptic non-NMDA-type glutamate receptors. At such NMJs, principal synapse formation proceeded and presynaptic active zones showed normal composition and ultrastructure as well as proper glutamate release. At the postsynaptic site, initial steps of molecular and structural assembly took place as well. However, growth of the nascent PSDs to mature size was inhibited, and proteins normally excluded from PSD membranes remained at these apparently immature sites. Intriguingly, synaptic transmission as well as glutamate binding to glutamate receptors appeared dispensable for synapse maturation. Thus, our data suggest that incorporation of non-NMDA-type glutamate receptors and likely their protein-protein interactions with additional PSD components triggers a conversion from an initial to a mature stage of PSD assembly. PMID- 17079655 TI - Rescuing qkV dysmyelination by a single isoform of the selective RNA-binding protein QKI. AB - Alternative splicing of the qkI transcript generates multiple isoforms of the selective RNA-binding protein QKI, which play key roles in controlling the homeostasis of their mRNA targets. QKI deficiency in oligodendrocytes of homozygous quakingviable (qkV/qkV) mutant mice results in severe hypomyelination, indicating the essential function of QKI in myelinogenesis. However, the molecular mechanisms by which QKI controls myelination remain elusive. We report here that QKI-6 is the most abundant isoform in brain and is preferentially reduced in the qkV/qkV mutant during normal myelinogenesis. To test whether QKI-6 is the predominant isoform responsible for advancing CNS myelination, we developed transgenic mice that express Flag-QKI-6 specifically in the oligodendroglia lineage, driven by the proteolipid protein (PLP) promoter. When introduced into the qkV/qkV mutant, the QKI-6 transgene rescues the severe tremor and hypomyelination phenotype. Electron microscopic studies further revealed that the Flag-QKI-6 transgene is sufficient for restoring compact myelin formation with normal lamellar periodicity and thickness. Interestingly, Flag-QKI-6 preferentially associates with the mRNA encoding the myelin basic protein (MBP) and rescues MBP expression from the beginning of myelinogenesis. In contrast, Flag-QKI-6 binds the PLP mRNA with lower efficiency and has a minimal impact on PLP expression until much later, when the expression level of QKI-6 in the transgenic animal significantly exceeds what is needed for normal myelination. Together, our results demonstrate that QKI-6 is the major isoform responsible for CNS myelination, which preferentially promotes MBP expression in oligodendrocytes. PMID- 17079656 TI - Abnormal function of C-fibers in patients with diabetic neuropathy. AB - The mechanisms underlying the development of painful and nonpainful neuropathy associated with diabetes mellitus are unclear. We have obtained microneurographic recordings from unmyelinated fibers in eight patients with diabetes mellitus, five with painful neuropathy, and three with neuropathy without pain. All eight patients had large-fiber neuropathy, and seven patients had pathological thermal thresholds in their feet, indicating the involvement of small-caliber nerve fibers. A total of 163 C-fibers were recorded at knee level from the common peroneal nerve in the patients (36-67 years old), and these were compared with 77 C-fibers from healthy controls (41-64 years old). The ratio of mechano-responsive to mechano-insensitive nociceptors was approximately 2:1 in the healthy controls, whereas in the patients, it was 1:2. In patients, a fairly large percentage of characterized fibers (12.5% in nonpainful and 18.9% in painful neuropathy) resembled mechano-responsive nociceptors that had lost their mechanical and heat responsiveness. Such fibers were rarely encountered in age-matched controls (3.2%). Afferent fibers with spontaneous activity or mechanical sensitization were found in both patient groups. We conclude that small-fiber neuropathy in diabetes affects receptive properties of nociceptors that leads to an impairment of mechano-responsive nociceptors. PMID- 17079657 TI - An acute effect of neuregulin 1 beta to suppress alpha 7-containing nicotinic acetylcholine receptors in hippocampal interneurons. AB - We examined rapid effects of neuregulin (NRG) on nicotinic acetylcholine (ACh) receptors in interneurons located in the stratum radiatum of the hippocampus. Two types of response were detected by whole-cell recordings after brief pulses of ACh. One type was a rapidly rising and falling (monophasic) current that was blocked by methyllycaconitine. The other type was a similar fast response followed by a more slowly rising and falling current. The slow component of the biphasic response was resistant to methyllycaconitine. Perfusion or local application with NRG 1beta rapidly decreased fast inward ACh currents. NRG 1beta had no effect on slow responses. NRG 1beta suppression was abolished by the ErbB tyrosine kinase inhibitor PD 158780 (4-[(3-bromophenyl) amino]-6-(methylamino) pyrido[3,4-d]pyridimine). The NRG 1beta effect was also inhibited by phalloidin and cytochalasin D. Furthermore, NRG 1beta decreased the number of surface Alexa Fluor 488 alpha-bungarotoxin binding sites. We believe that the NRG 1beta-induced inhibition of ACh currents is because of receptor internalization trigged by protein tyrosine phosphorylation. Significantly, fast nicotinic EPSCs evoked in the presence of muscarinic, ionotropic glutamate, and GABA receptors antagonists were also reduced by NRG 1beta. Thus, short-term as well as long-term effects of NRG must be taken into consideration in studies of ACh receptor-mediated synaptic efficacy in the CNS. PMID- 17079658 TI - Experience-dependent eye movements, awareness, and hippocampus-dependent memory. AB - We asked what kind of memory is operating when eye movements change as the result of experience. Participants viewed scenes that were either novel, repeated, or manipulated (i.e., a change was introduced in one region of the scene). Eye movements differed depending on the past viewing history of each scene. Participants made fewer fixations and sampled fewer regions when scenes were repeated than when scenes were novel. When scenes were altered, participants made more fixations in the altered region, spent more time looking at the altered region, and made more transitions into and out of the altered region than in unchanged (matched) regions in the repeated scenes. Importantly, these effects occurred only when individuals were aware that a change had occurred. Participants who were unaware that the scene had been altered looked at the changed scenes in the same way that they looked at repeated scenes. Thus, there was no indication that eye movements could reveal an unaware (unconscious) form of memory. Instead, eye movements reflected conscious memory of whether the scene was repeated or manipulated. The findings were the same when awareness was assessed after viewing all the scenes (experiment 1) and when awareness was assessed after each scene was presented (experiment 2). In experiment 3, memory impaired patients with damage limited to the hippocampus were impaired at deciding whether scenes were novel, repeated, or manipulated. Thus, the ability to consciously recollect recent encounters with scenes reflects a form of hippocampus-dependent memory. The findings show that experience-dependent eye movements in response to altered scenes reflect conscious, declarative memory, and they support the link between aware memory, declarative memory, and hippocampus-dependent memory. PMID- 17079659 TI - Modulation of dorsolateral prefrontal delay activity during self-organized behavior. AB - The regulation of cognitive activity relies on the flexibility of prefrontal cortex functions. To study this mechanism we compared monkey dorsolateral prefrontal activity in two different spatial cognitive tasks: a delayed response task and a self-organized problem-solving task. The latter included two periods, a search by trial and error for a correct response, and a repetition of the response once discovered. We show that (1) delay activity involved in the delayed task also participates in self-generated responses during the problem-solving task and keeps the same location preference, and (2) the amplitude of firing and the strength of spatial selectivity vary with task requirement, even within search periods while approaching the correct response. This variation is dissociated from pure reward probability, but may have a link with uncertainty because the selectivity dropped when reward predictability was maximal. Overall, we show that spatially tuned delay activity of prefrontal neurons reflects the varying level of engagement in control between different spatial cognitive tasks and during self-organized behavior. PMID- 17079660 TI - Corticotropin-releasing factor within the central nucleus of the amygdala mediates enhanced ethanol self-administration in withdrawn, ethanol-dependent rats. AB - Alcohol dependence is characterized by excessive consumption, loss of control over intake, and the presence of a withdrawal syndrome, including both motivational and physical symptoms. The motivational symptoms, including anxiety, have been hypothesized to be important factors eliciting excessive drinking during abstinence. Previous work has shown that ethanol-dependent rats also display enhanced anxiety-like behaviors and enhanced ethanol self-administration during withdrawal, likely resulting from dysregulation of brain corticotropin releasing factor (CRF) stress systems. The present study was designed to explore the brain sites within the extended amygdala [central nucleus of the amygdala (CeA), lateral bed nucleus of the stria terminalis (BNST), and nucleus accumbens shell (NAcSh)] that mediate the increased ethanol self-administration observed during withdrawal. Ethanol-dependent animals showed an increase in ethanol self administration after acute withdrawal relative to nondependent controls. The CRF antagonist D-Phe-CRF(12-41) ([D-Phe(12),Nle(21,38),C alpha MeLeu(37)]-rCRF(12 41)) was administered into the CeA, lateral BNST, or NAcSh of acute-withdrawn dependent and nondependent rats. Administered into the CeA, the antagonist reduced ethanol self-administration in dependent animals, with no effect in nondependent animals. Administration of D-Phe-CRF(12-41) into the lateral BNST and NAcSh was without effect on ethanol self-administration in dependent and nondependent animals. At the same time point of withdrawal, there was a decrease in CRF immunoreactivity within the CeA, suggesting an increased extracellular release of CRF during withdrawal. There was no change in CRF immunoreactivity in the BNST or NAcSh. These results indicate that CRF, specifically within the CeA, plays a role in mediating excessive ethanol consumption in ethanol-dependent animals. PMID- 17079661 TI - Proteasome inhibition triggers activity-dependent increase in the size of the recycling vesicle pool in cultured hippocampal neurons. AB - The ubiquitin proteasome system, generally known for its function in protein degradation, also appears to play an important role in regulating membrane trafficking. A role for the proteasome in regulating presynaptic release and vesicle trafficking has been proposed for invertebrates, but it remains to be tested in mammalian presynaptic terminals. We used the fluorescent styrylpyridinium dye FM4-64 to visualize changes in the recycling pool of vesicles in hippocampal culture under pharmacological inhibition of the proteasome. We found that a 2 h inhibition increases the recycling pool of vesicles by 76%, with no change in the rate or total amount of dye release. Interestingly, enhancement did not depend on protein synthesis but did depend on synaptic activity; blocking action potentials during proteasome inhibition abolished the effect whereas increasing neuronal activity accelerated the effect with an increased recycling pool evident after 15 min. We propose that the proteasome acts as a negative-feedback regulator of synaptic transmission, possibly serving a homeostatic role. PMID- 17079662 TI - Enhancing GABA(A) receptor alpha 1 subunit levels in hippocampal dentate gyrus inhibits epilepsy development in an animal model of temporal lobe epilepsy. AB - Differential expression of GABA(A) receptor (GABR) subunits has been demonstrated in hippocampus from patients and animals with temporal lobe epilepsy (TLE), but whether these changes are important for epileptogenesis remains unknown. Previous studies in the adult rat pilocarpine model of TLE found reduced expression of GABR alpha1 subunits and increased expression of alpha4 subunits in dentate gyrus (DG) of epileptic rats compared with controls. To investigate whether this altered subunit expression is a critical determinant of spontaneous seizure development, we used adeno-associated virus type 2 containing the alpha4 subunit gene (GABRA4) promoter to drive transgene expression in DG after status epilepticus (SE). This novel use of a condition-dependent promoter upregulated after SE successfully increased expression of GABR alpha1 subunit mRNA and protein in DG at 1-2 weeks after SE. Enhanced alpha1 expression in DG resulted in a threefold increase in mean seizure-free time after SE and a 60% decrease in the number of rats developing epilepsy (recurrent spontaneous seizures) in the first 4 weeks after SE. These findings provide the first direct evidence that altering GABR subunit expression can affect the development of epilepsy and suggest that alpha1 subunit levels are important determinants of inhibitory function in hippocampus. PMID- 17079663 TI - Enhanced performance with brain stimulation: attentional shift or visual cue? AB - The premotor theory of visual spatial attention proposes that the same brain activity that prepares for saccades to one part of the visual field also facilitates visual processing at that same region of the visual field. Strong support comes from improvements in performance by electrical stimulation of presaccadic areas, including the frontal eye field and superior colliculus (SC). Interpretations of these stimulation experiments are hampered by the possibility that stimulation might be producing an internal visual flash or phosphene that attracts attention as a real flash would. We tested this phosphene hypothesis in the SC by comparing the effect of interchanging real visual stimuli and electrical stimulation. We first presented a veridical visual cue at the time SC stimulation improved performance; if a phosphene improved performance at this time, a real cue should do so in the same manner, but it did not. We then changed the time of SC visual-motor stimulation to when we ordinarily presented the veridical visual cue, and failed to improve performance. Last, we shifted the site of SC stimulation from the visual-motor neurons of the SC intermediate layers to the visual neurons of the superficial layers to determine whether stimulating visual neurons produced a larger improvement in performance, but it did not. Our experiments provide evidence that a phosphene is not responsible for the shift of attention that follows SC stimulation. This added evidence of a direct shift of attention is consistent with a key role of the SC in the premotor theory of attention. PMID- 17079664 TI - The transcriptional coactivator Querkopf controls adult neurogenesis. AB - The adult mammalian brain maintains populations of neural stem cells within discrete proliferative zones. Understanding of the molecular mechanisms regulating adult neural stem cell function is limited. Here, we show that MYST family histone acetyltransferase Querkopf (Qkf, Myst4, Morf)-deficient mice have cumulative defects in adult neurogenesis in vivo, resulting in declining numbers of olfactory bulb interneurons, a population of neurons produced in large numbers during adulthood. Qkf-deficient mice have fewer neural stem cells and fewer migrating neuroblasts in the rostral migratory stream. Qkf gene expression is strong in the neurogenic subventricular zone. A population enriched in multipotent cells can be isolated from this region on the basis of Qkf gene expression. Neural stem cells/progenitor cells isolated from Qkf mutant mice exhibited a reduced self-renewal capacity and a reduced ability to produce differentiated neurons. Together, our data show that Qkf is essential for normal adult neurogenesis. PMID- 17079665 TI - Adenosine A2A receptors in bone marrow-derived cells but not in forebrain neurons are important contributors to 3-nitropropionic acid-induced striatal damage as revealed by cell-type-selective inactivation. AB - Endogenous adenosine acting at the adenosine A2A receptor (A2AR) can modify brain injury in a variety of neurological disorder models. However, both A2AR activation and inactivation have been shown to be neuroprotective in different situations, raising the intriguing possibility that A2ARs in distinct cellular elements may have different and even opposing effects. In this study, we developed three novel transgenic models to dissect out cell-type-specific actions of A2ARs on striatal damage by the mitochondrial toxin 3-nitropropionic acid (3 NP). Whereas global inactivation of A2ARs exacerbated 3-NP-induced neurological deficit behaviors and striatal damage, selective inactivation of A2ARs in forebrain neurons (using the Cre/loxP strategy) did not affect neurological deficit or striatal damage after the acute systemic treatment of 3-NP and intrastriatal injection of malonate. However, selective inactivation of A2ARs in bone marrow-derived cells (BMDCs) by transplanting bone marrow cells from global A2AR knock-out (KO) mice into wild-type C57BL/6 mice produced a similar phenotype of global A2AR KO mice, i.e., exacerbation of 3-NP-induced striatal damage. Thus, cell-type-selective inactivation of A2ARs reveals that A2ARs in BMDCs but not in forebrain neurons are an important contributor to striatal damage induced by mitochondrial dysfunction. PMID- 17079666 TI - Choosing the lesser of two evils, the better of two goods: specifying the roles of ventromedial prefrontal cortex and dorsal anterior cingulate in object choice. AB - The ventromedial prefrontal cortex (vmPFC) and dorsal anterior cingulate cortices (ACd) are considered important for reward-based decision making. However, work distinguishing their individual functional contributions has only begun. One aspect of decision making that has received little attention is that making the right choice often translates to making the better choice. Thus, response choice often occurs in situations where both options are desirable (e.g., choosing between mousse au chocolat or creme caramel cheesecake from a menu) or, alternatively, in situations where both options are undesirable. Moreover, response choice is easier when the reinforcements associated with the objects are far apart, rather than close together, in value. We used functional magnetic resonance imaging to delineate the functional roles of the vmPFC and ACd by investigating these two aspects of decision making: (1) decision form (i.e., choosing between two objects to gain the greater reward or the lesser punishment), and (2) between-object reinforcement distance (i.e., the difference in reinforcements associated with the two objects). Blood oxygen level-dependent (BOLD) responses within the ACd and vmPFC were both related to decision form but differentially. Whereas ACd showed greater responses when deciding between objects to gain the lesser punishment, vmPFC showed greater responses when deciding between objects to gain the greater reward. Moreover, vmPFC was sensitive to reinforcement expectations associated with both the chosen and the forgone choice. In contrast, BOLD responses within ACd, but not vmPFC, related to between-object reinforcement distance, increasing as the distance between the reinforcements of the two objects decreased. These data are interpreted with reference to models of ACd and vmPFC functioning. PMID- 17079667 TI - Increased citrullination of histone H3 in multiple sclerosis brain and animal models of demyelination: a role for tumor necrosis factor-induced peptidylarginine deiminase 4 translocation. AB - Modification of arginine residues by citrullination is catalyzed by peptidylarginine deiminases (PADs), of which five are known, generating irreversible protein structural modifications. We have shown previously that enhanced citrullination of myelin basic protein contributed to destabilization of the myelin membrane in the CNS of multiple sclerosis (MS) patients. We now report increased citrullination of nucleosomal histones by PAD4 in normal-appearing white matter (NAWM) of MS patients and in animal models of demyelination. Histone citrullination was attributable to increased levels and activity of nuclear PAD4. PAD4 translocation into the nucleus was attributable to elevated tumor necrosis factor-alpha (TNF-alpha) protein. The elevated TNF-alpha in MS NAWM was not associated with CD3+ or CD8+ lymphocytes, nor was it associated with CD68+ microglia/macrophages. GFAP, a measure of astrocytosis, was the only cytological marker that was consistently elevated in the MS NAWM, suggesting that TNF-alpha may have been derived from astrocytes. In cell cultures of mouse and human oligodendroglial cell lines, PAD4 was predominantly cytosolic but TNF-alpha treatment induced its nuclear translocation. To address the involvement of TNF alpha in targeting PAD4 to the nucleus, we found that transgenic mice overexpressing TNF-alpha also had increased levels of citrullinated histones and elevated nuclear PAD4 before demyelination. In conclusion, high citrullination of histones consequent to PAD4 nuclear translocation is part of the process that leads to irreversible changes in oligodendrocytes and may contribute to apoptosis of oligodendrocytes in MS. PMID- 17079668 TI - Absence of tumor necrosis factor-alpha does not affect motor neuron disease caused by superoxide dismutase 1 mutations. AB - An increase in the expression of the proinflammatory cytokine tumor necrosis factor alpha (TNF-alpha) has been observed in patients with amyotrophic lateral sclerosis (ALS) and in the mice models of the disease. TNF-alpha is a potent activator of macrophages and microglia and, under certain conditions, can induce or exacerbate neuronal cell death. Here, we assessed the contribution of TNF alpha in motor neuron disease in mice overexpressing mutant superoxide dismutase 1 (SOD1) genes linked to familial ALS. This was accomplished by the generation of mice expressing SOD1(G37R) or SOD1(G93A) mutants in the context of TNF-alpha gene knock out. Surprisingly, the absence of TNF-alpha did not affect the lifespan or the extent of motor neuron loss in SOD1 transgenic mice. These results provide compelling evidence indicating that TNF-alpha does not directly contribute to motor neuron degeneration caused by SOD1 mutations. PMID- 17079669 TI - Weakened center-surround interactions in visual motion processing in schizophrenia. AB - Schizophrenia is often accompanied by a range of visual perception deficits, with many involving impairments in motion perception. The presence of perceptual abnormalities may impair neural processes that depend on normal visual analysis, which in turn may affect overall functioning in dynamic visual environments. Here, we examine the integrity of suppressive center-surround mechanisms in motion perception of schizophrenic patients. Center-surround suppression has been implicated in a range of visual functions, including figure-ground segregation and pursuit eye movements, visual functions that are impaired in schizophrenia. In control subjects, evidence of center-surround suppression is found in a reduced ability to perceive motion of a high-contrast stimulus as its size increases. This counterintuitive finding is likely a perceptual correlate of center-surround mechanisms in cortical area MT. We now show that schizophrenic patients exhibit abnormally weak center-surround suppression in motion, an abnormality that is most pronounced in patients with severe negative symptoms. Interestingly, patients with the weakest surround suppression outperformed control subjects in motion discriminations of large high-contrast stimuli. This enhanced motion perception of large high-contrast stimuli is consistent with an MT abnormality in schizophrenia and has a potential to disrupt smooth pursuit eye movements and other visual functions that depend on unimpaired center-surround interactions in motion. PMID- 17079670 TI - Fusion of microglia with pyramidal neurons after retroviral infection. AB - The neurogenic potential of the postnatal neocortex has not been tested previously with a combination of both retroviral and bromodeoxyuridine (BrdU) labeling. Here we report that injections of enhanced green fluorescent protein (eGFP) retrovirus into 134 postnatal rats resulted in GFP labeling of 642 pyramidal neurons in neocortex. GFP-labeled neocortical pyramidal neurons, however, unlike GFP-labeled glia, did not incorporate BrdU. Closer inspection of retrovirally labeled neurons revealed microglia fused to the apical dendrites of labeled pyramidal neurons. Retroviral infection of mixed cultures of cortical neurons and glia confirmed the presence of specific neuronal-microglial fusions. Microglia did not fuse to other glial cell types, and cultures not treated with retrovirus lacked microglial-neuronal fusion. Furthermore, activation of microglia by lipopolysaccharide greatly increased the virally induced fusion of microglia to neurons in culture. These results indicate a novel form of specific cell fusion between neuronal dendrites and microglia and further illustrate the need for caution when interpreting evidence for neuronogenesis in the postnatal brain. PMID- 17079671 TI - Transgenic rescue of ataxia mice with neuronal-specific expression of ubiquitin specific protease 14. AB - The ataxia mutation (axJ) is a recessive neurological mutation that results in reduced growth, ataxia, and hindlimb muscle wasting in mice. The axJ gene encodes ubiquitin-specific protease 14 (Usp14), a deubiquitinating enzyme (DUB) that associates with the proteasome via its ubiquitin-like (Ubl) domain and is involved in processing ubiquitin chains. Analysis of Usp14 gene products demonstrated that Usp14 undergoes alternative pre-mRNA splicing to produce a full length form of Usp14 that is capable of binding proteasomes and a form that contains a deletion in the Ubl domain. The full-length form of Usp14 is the only form that appears to be reduced in the axJ mice. Transgenic rescue of the axJ mice with neuronal-specific expression of Usp14 demonstrated that the full-length form of Usp14 was sufficient to restore viability and motor system function to the axJ mice. Biochemical analysis showed that the ubiquitin hydrolyase activity of this form of Usp14 is dependent on the presence of proteasomes, and neuronal expression of full-length Usp14 was able to restore the levels of monomeric ubiquitin in the brains of axJ mice. However, the axJ-rescued mice still displayed the Purkinje cell axonal swellings that are seen in the axJ mice, indicating that this cerebellar alteration is not the primary cause of the axJ movement disorders. These results show that the motor defects observed in the axJ mice are attributable to a neuropathic disease rather than to a muscular disorder and suggest that changes in proteasomal function may contribute to neurological dysfunction in the axJ mice. PMID- 17079672 TI - Estimate of the chloride concentration in a central glutamatergic terminal: a gramicidin perforated-patch study on the calyx of Held. AB - The function of presynaptic terminals is regulated by intracellular Cl-, the levels of which modify vesicular endocytosis and transmitter refilling and mediate the effects of presynaptic ligand-gated Cl- channels. Nevertheless, the concentration of Cl- in a central nerve terminal is unknown, and it is unclear whether terminals can regulate Cl- independently of the soma. Using perforated patch recording in a mammalian synapse, we found that terminals accumulate Cl- up to 21 mm, between four and five times higher than in their parent cell bodies. Changing [Cl-] did not alter vesicular glutamate content in intact terminals, unlike in vitro experiments. Thus, glutamatergic terminals maintain an elevated Cl- concentration without compromising synaptic transmission. PMID- 17079673 TI - Increased T cell recruitment to the CNS after amyloid beta 1-42 immunization in Alzheimer's mice overproducing transforming growth factor-beta 1. AB - Immunotherapy targeting the amyloid beta (Abeta) peptide is a novel therapy under investigation for the treatment of Alzheimer's disease (AD). A clinical trial using Abeta(1-42) (AN1792) as the immunogen was halted as a result of development of meningoencephalitis in a small number of patients. The cytokine TGF-beta1 is a key modulator of immune responses that is increased in the brain in AD. We show here that local overexpression of TGF-beta1 in the brain increases both meningeal and parenchymal T lymphocyte number. Furthermore, TGF-beta1 overexpression in a mouse model for AD [amyloid precursor protein (APP) mice] leads to development of additional T cell infiltrates when mice were immunized at a young but not old age with AN1792. Notably, only mice overproducing both Abeta (APP mice) and TGF-beta1 experienced a rise in T lymphocyte number after immunization. One-third of infiltrating T cells were CD4 positive. We did not observe significant differences in B lymphocyte numbers in any of the genotypes or treatment groups. These results demonstrate that TGF-beta1 overproduction in the brain can promote T cell infiltration, in particular after Abeta(1-42) immunization. Likewise, levels of TGF-beta1 or other immune factors in brains of AD patients may influence the response to Abeta(1-42) immunization. PMID- 17079674 TI - Noradrenergic inputs to the bed nucleus of the stria terminalis and paraventricular nucleus of the hypothalamus underlie hypothalamic-pituitary adrenal axis but not hypophagic or conditioned avoidance responses to systemic yohimbine. AB - The alpha2 adrenoceptor antagonist yohimbine (YO) increases transmitter release from adrenergic/noradrenergic (NA) neurons. Systemic YO activates the hypothalamic-pituitary-adrenal (HPA) axis, inhibits feeding, and supports conditioned flavor avoidance (CFA) in rats. To determine whether these effects require NA inputs to the bed nucleus of the stria terminalis (BNST), vehicle or saporin toxin conjugated to an antibody against dopamine beta hydroxylase (DSAP) was microinjected bilaterally into the BNST to remove its NA inputs. Subsequent tests failed to reveal any lesion effect on the ability of YO (5.0 mg/kg, i.p.) to inhibit food intake or to support CFA. Conversely, HPA axis responses to YO were significantly blunted in DSAP rats. In a terminal experiment, DSAP and control rats were perfused 90-120 min after intraperitoneal injection of YO or vehicle. Brains were processed to reveal Fos immunolabeling and lesion extent. NA fibers were markedly depleted in the BNST and medial parvocellular paraventricular hypothalamus (PVNmp) in DSAP rats, evidence for collateralized NA inputs to these regions. DSAP rats displayed significant loss of caudal medullary NA neurons, and markedly blunted Fos activation in the BNST and in corticotropin releasing hormone-positive PVNmp neurons after YO. We conclude that a population of medullary NA neurons provides collateral inputs to the BNST and PVNmp, and that these inputs contribute importantly to Fos expression and HPA axis activation after YO treatment. Conversely, NA-mediated activation of BNST and PVNmp neurons is unnecessary for YO to inhibit food intake or support CFA, evidence for the sufficiency of other intact neural pathways in mediating those effects. PMID- 17079675 TI - Emotional modulation of pain: is it the sensation or what we recall? AB - Emotions modulate pain perception, although the mechanisms underlying this phenomenon remain unclear. In this study, we show that intensity reports significantly increased when painful stimuli were concomitant to images showing human pain, whereas pictures with identical emotional values but without somatic content failed to modulate pain. Early somatosensory responses (<200 ms) remained unmodified by emotions. Conversely, late responses showed a significant enhancement associated with increased pain ratings, localized to the right prefrontal, right temporo-occipital junction, and right temporal pole. In contrast to selective attention, which enhances pain ratings by increasing sensory gain, emotions triggered by seeing other people's pain did not alter processing in SI-SII (primary and second somatosensory areas), but may have biased the transfer to, and the representation of pain in short-term memory buffers (prefrontal), as well as the affective assignment to this representation (temporal pole). Memory encoding and recall, rather than sensory processing, appear to be modulated by empathy with others' physical suffering. PMID- 17079676 TI - Panoramic measurements of the apex of the cochlea. AB - Our understanding of cochlear mechanics is impeded by the lack of truly panoramic data. Sensitive mechanical measurements cover only a narrow cochlear region, mostly in the base. The global spatiotemporal pattern of vibrations along the cochlea cannot be inferred from such local measurements but is often extrapolated beyond the measurement spot under the assumption of scaling invariance. Auditory nerve responses give an alternative window on the entire cochlea, but traditional techniques do not allow recovery of the effective vibration pattern. We developed a new analysis technique to measure cochlear amplitude and phase transfer of fibers with characteristic frequencies <5 kHz. Data from six cats yielded panoramic phase profiles along the apex of the cochlea for an approximately 5 octave range of stimulus frequencies. All profiles accumulated systematic phase lags from base to apex. Phase accumulation was not gradual but showed a two segment character: a steep segment (slow propagation) around the characteristic position of the stimulus, and a shallow segment (fast propagation) basal to it. The transition between the segments occurred in a narrow region and was smooth. Wavelength near characteristic position decreased from approximately 3.5 to approximately 1 mm for frequencies from 200 to 4000 Hz, corresponding to phase velocities of approximately 0.5 to approximately 5 m/s. The accumulated phase lag between the eardrum and characteristic position varied from approximately 1 cycle at 200 Hz to approximately 2.5 cycle at 4 kHz, invalidating scaling invariance. The generic character of our analysis technique and its success in solving the difficult problem of reconstructing the effective sensory input from neural recordings suggest its wider application as a powerful alternative to customary system analysis techniques. PMID- 17079677 TI - Ataxin-3 represses transcription via chromatin binding, interaction with histone deacetylase 3, and histone deacetylation. AB - Ataxin-3 (AT3), the disease protein in spinocerebellar ataxia type 3 (SCA3), has been associated with the ubiquitin-proteasome system and transcriptional regulation. Here we report that normal AT3 binds to target DNA sequences in specific chromatin regions of the matrix metalloproteinase-2 (MMP-2) gene promoter and represses transcription by recruitment of the histone deacetylase 3 (HDAC3), the nuclear receptor corepressor (NCoR), and deacetylation of histones bound to the promoter. Both normal and expanded AT3 physiologically interacted with HDAC3 and NCoR in a SCA3 cell model and human pons tissue; however, normal AT3-containing protein complexes showed increased histone deacetylase activity, whereas expanded AT3-containing complexes had reduced deacetylase activity. Consistently, histone analyses revealed an increased acetylation of total histone H3 in expanded AT3-expressing cells and human SCA3 pons. Expanded AT3 lost the repressor function and displayed altered DNA/chromatin binding that was not associated with recruitment of HDAC3, NCoR, and deacetylation of the promoter, allowing aberrant MMP-2 transcription via the transcription factor GATA-2. For transcriptional repression normal AT3 cooperates with HDAC3 and requires its intact ubiquitin-interacting motifs (UIMs), whereas aberrant transcriptional activation by expanded AT3 is independent of the UIMs but requires the catalytic cysteine of the ubiquitin protease domain. These findings demonstrate that normal AT3 binds target promoter regions and represses transcription of a GATA-2 dependent target gene via formation of histone-deacetylating repressor complexes requiring its UIM-associated function. Expanded AT3 aberrantly activates transcription via its catalytic site and loses the ability to form deacetylating repressor complexes on target chromatin regions. PMID- 17079678 TI - The dynamic distribution of TrkB receptors before, during, and after synapse formation between cortical neurons. AB - Although brain-derived neurotrophic factor (BDNF) potently regulates neuronal connectivity in the developing CNS, the mechanism by which BDNF influences the formation and/or maintenance of glutamatergic synapses remains unknown. Details about the subcellular localization of the BDNF receptor, TrkB, relative to synaptic and nonsynaptic proteins on excitatory neurons should provide insight into how BDNF might exert its effects during synapse formation. Here, we investigated the subcellular localization of tyrosine kinase receptor B (TrkB) relative to synaptic vesicle-associated proteins and NMDA receptors using immunocytochemistry, confocal microscopy, and time-lapse imaging in dissociated cultures of cortical neurons before, during, and after the peak of synapse formation. We find that TrkB is present in puncta on the surface and intracellularly in both dendrites and axons throughout development. Before synapse formation, some TrkB puncta in dendrites colocalize with NMDA receptors, and almost all TrkB puncta in axons colocalize with synaptic vesicle proteins. Clusters of TrkB fused to the enhanced green fluorescent protein (TrkB-EGFP) are highly mobile in both axons and dendrites. In axons, TrkB-EGFP dynamics are almost identical to vesicle-associated protein (VAMP2-EGFP), and these proteins are often transported together. Finally, surface TrkB is found in structures that actively participate in synapse formation: axonal growth cones and dendritic filopodia. Over time, surface TrkB becomes enriched at glutamatergic synapses, which contain both catalytic and truncated TrkB. These results suggest that TrkB is in the right place at the right time to play a direct role in the formation of glutamatergic synapses between cortical neurons. PMID- 17079679 TI - Anterolateral prefrontal cortex mediates the analgesic effect of expected and perceived control over pain. AB - Perceived control attenuates pain and pain-directed anxiety, possibly because it changes the emotional appraisal of pain. We examined whether brain areas associated with voluntary reappraisal of emotional experiences also mediate the analgesic effect of perceived control over pain. Using functional magnetic resonance imaging, we compared self-controlled noxious stimuli with physically identical stimuli that were externally controlled. Self-controlled stimulation was accompanied by less pain and anxiety and higher activation in dorsal anterior cingulate (dACC), right dorsolateral, and bilateral anterolateral prefrontal (alPFC) cortices. Activation in dACC and right alPFC was negatively correlated with pain intensity ratings. For externally controlled pain, activation in right alPFC was inversely correlated with the participants' general belief to have control over their lives. Our results are consistent with a reappraisal view of control and suggest that the analgesic effect of perceived control relies on activation of right alPFC. Failure to activate right alPFC may explain the maladaptive effects of strong general control beliefs during uncontrollable pain. PMID- 17079680 TI - Salmonella produces an O-antigen capsule regulated by AgfD and important for environmental persistence. AB - In this study, we show that Salmonella produces an O-antigen capsule coregulated with the fimbria- and cellulose-associated extracellular matrix. Structural analysis of purified Salmonella extracellular polysaccharides yielded predominantly a repeating oligosaccharide unit similar to that of Salmonella enterica serovar Enteritidis lipopolysaccharide O antigen with some modifications. Putative carbohydrate transport and regulatory operons important for capsule assembly and translocation, designated yihU-yshA and yihVW, were identified by screening a random transposon library with immune serum generated to the capsule. The absence of capsule was confirmed by generating various isogenic Deltayih mutants, where yihQ and yihO were shown to be important in capsule assembly and translocation. Luciferase-based expression studies showed that AgfD regulates the yih operons in coordination with extracellular matrix genes coding for thin aggregative fimbriae and cellulose. Although the capsule did not appear to be important for multicellular behavior, we demonstrate that it was important for survival during desiccation stress. Since the yih genes are conserved in salmonellae and the O-antigen capsule was important for environmental persistence, the formation of this surface structure may represent a conserved survival strategy. PMID- 17079681 TI - Influence of the two-component system SaeRS on global gene expression in two different Staphylococcus aureus strains. AB - The two-component system SaeRS consisting of the histidin kinase SaeS and the response regulator SaeR is known to act on virulence gene expression in Staphylococcus aureus. In order to get a more comprehensive picture on SaeR regulated genes, we studied the contribution of the two-component system on global gene expression by using both the proteomic and transcriptomic approach. Altogether, a loss of SaeRS resulted in a decreased amount of at least 17 extracellular proteins and two cell surface-associated proteins, among them several important virulence factors such as HlgA, HlgB, HlgC, LukF, and LukM. SaeRS activates the expression of these genes at the transcriptional level. The amount of the five proteins Aur, SspA, SsaA, Plc, and GlpQ was negatively influenced by SaeRS. However, the transcription of the corresponding genes was not affected by the two-component system. SaeRS had also no measurable influence on the transcription of the regulatory genes agr, sarA, arlRS, and sigB that contribute to the regulation of SaeRS-dependent virulence factors identified in this investigation. Our results clearly show that SaeRS is strongly involved in the tight temporal control of virulence factor expression in S. aureus. Its precise role within the regulatory network remains to be determined. PMID- 17079682 TI - Mammalian sirtuins--emerging roles in physiology, aging, and calorie restriction. AB - Sir2 is an NAD-dependent deacetylase that connects metabolism with longevity in yeast, worms and flies. Mammals contain seven homologs of yeast Sir2, SIRT1-7. Here, we review recent findings demonstrating the role of these mammalian sirtuins as regulators of physiology, calorie restriction, and aging. The current findings sharpen our understanding of sirtuins as potential pharmacological targets to treat the major diseases of aging. PMID- 17079683 TI - Phosphorylation and functions of the RNA polymerase II CTD. AB - The C-terminal repeat domain (CTD), an unusual extension appended to the C terminus of the largest subunit of RNA polymerase II, serves as a flexible binding scaffold for numerous nuclear factors; which factors bind is determined by the phosphorylation patterns on the CTD repeats. Changes in phosphorylation patterns, as polymerase transcribes a gene, are thought to orchestrate the association of different sets of factors with the transcriptase and strongly influence functional organization of the nucleus. In this review we appraise what is known, and what is not known, about patterns of phosphorylation on the CTD of RNA polymerases II at the beginning, the middle, and the end of genes; the proposal that doubly phosphorylated repeats are present on elongating polymerase is explored. We discuss briefly proteins known to associate with the phosphorylated CTD at the beginning and ends of genes; we explore in more detail proteins that are recruited to the body of genes, the diversity of their functions, and the potential consequences of tethering these functions to elongating RNA polymerase II. We also discuss accumulating structural information on phosphoCTD-binding proteins and how it illustrates the variety of binding domains and interaction modes, emphasizing the structural flexibility of the CTD. We end with a number of open questions that highlight the extent of what remains to be learned about the phosphorylation and functions of the CTD. PMID- 17079684 TI - DDB1 functions as a linker to recruit receptor WD40 proteins to CUL4-ROC1 ubiquitin ligases. AB - Cullins assemble the largest family of ubiquitin ligases by binding with ROC1 and various substrate receptors. CUL4 function is linked with many cellular processes, but its substrate-recruiting mechanism remains elusive. We identified a protein motif, the DWD box (DDB1-binding WD40 protein), and demonstrated the binding of 15 DWD proteins with DDB1-CUL4A. We provide evidence supporting the critical function of the DWD box and DDB1's role as the linker mediating DWD protein association with CUL4A. A database search predicts that about one-third of WD40 proteins, 90 in humans, contain the DWD box, suggesting a potentially large number of DWD-DDB1-CUL4-ROC1 E3 ligases. PMID- 17079685 TI - Insulin-like signaling and the neural circuit for integrative behavior in C. elegans. AB - Caenorhabditis elegans exhibits a food-associated behavior that is modulated by the past cultivation temperature. Mutations in INS-1, the homolog of human insulin, caused the defect in this integrative behavior. Mutations in DAF 2/insulin receptor and AGE-1/phosphatidylinositol 3 (PI-3)-kinase partially suppressed the defect of ins-1 mutants, and a mutation in DAF-16, a forkhead-type transcriptional factor, caused a weak defect. In addition, mutations in the secretory protein HEN-1 showed synergistic effects with INS-1. Expression of AGE 1 in any of the three interneurons, AIY, AIZ, or RIA, rescued the defect characteristic of age-1 mutants. Calcium imaging revealed that starvation induced INS-1-mediated down-regulation of AIZ activity. Our results suggest that INS-1, in cooperation with HEN-1, antagonizes the DAF-2 insulin-like signaling pathway to modulate interneuron activity required for food-associated integrative behavior. PMID- 17079686 TI - Condensin is required for chromosome arm cohesion during mitosis. AB - We describe a novel requirement for the condensin complex in sister chromatid cohesion in Saccharomyces cerevisiae. Strikingly, condensin-dependent cohesion can be distinguished from cohesin-based pairing by a number of criteria. First, condensin is required to maintain cohesion at several chromosomal arm sites but, in contrast to cohesin, is not required at either centromere or telomere-proximal loci. Second, condensin-dependent interlinks are established during mitosis independently of DNA replication and are reversible within a single cell cycle. Third, the loss of condensin-dependent linkages occurs without affecting cohesin levels at the separated URA3 locus. We propose that, during mitosis, robust sister chromatid cohesion along chromosome arms requires both condensinand cohesin-dependent mechanisms, which function independently of each other. We discuss the implications of our results for current models of sister chromatid cohesion. PMID- 17079687 TI - The RNA silencing endonuclease Argonaute 2 mediates specific antiviral immunity in Drosophila melanogaster. AB - Most organisms have evolved defense mechanisms to protect themselves from viruses and other pathogens. Arthropods lack the protein-based adaptive immune response found in vertebrates. Here we show that the central catalytic component of the RNA-induced silencing complex (RISC), the nuclease Argonaute 2 (Ago-2), is essential for antiviral defense in adult Drosophila melanogaster. Ago-2-defective flies are hypersensitive to infection with a major fruit fly pathogen, Drosophila C virus (DCV), and with Cricket Paralysis virus (CrPV). Increased mortality in ago-2 mutant flies was associated with a dramatic increase in viral RNA accumulation and virus titers. The physiological significance of this antiviral mechanism is underscored by our finding that DCV encodes a potent suppressor of RNA interference (RNAi). This suppressor binds long double-stranded RNA (dsRNA) and inhibits Dicer-2-mediated processing of dsRNA into short interfering RNA (siRNA), but does not bind short siRNAs or disrupt the microRNA (miRNA) pathway. Based on these results we propose that RNAi is a major antiviral immune defense mechanism in Drosophila. PMID- 17079688 TI - The order of expression of transcription factors directs hierarchical specification of hematopoietic lineages. AB - The mechanism of lineage specification in multipotent stem cells has not been fully understood. We recently isolated progenitors with the eosinophil, basophil, or mast cell lineage potential, all of which originate from granulocyte/monocyte progenitors (GMPs). By using these prospectively purified progenitors, we show here that the expression timing of GATA-2 and CCAAT enhancer-binding protein alpha (C/EBPalpha) can differentially control their lineage commitment. The expression of GATA-2 instructed C/EBPalpha-expressing GMPs to commit exclusively into the eosinophil lineage, while it induced basophil and/or mast cell lineage commitment if C/EBPalpha was suppressed at the GMP stage. Furthermore, simply by switching the order of C/EBPalpha and GATA-2 transduction, even lymphoid committed progenitors recaptured these developmental processes to be reprogrammed into each of these lineages. We propose that the order of expression of key transcription factors is critical for their interplay to selectively drive lineage specification programs, by which stem cells could generate multiple lineage cells in a hierarchical manner. PMID- 17079689 TI - Canonical notch signaling functions as a commitment switch in the epidermal lineage. AB - Mammalian epidermis consists of a basal layer of proliferative progenitors that gives rise to multiple differentiating layers to provide a waterproof envelope covering the skin surface. To accomplish this, progenitor cells must detach from the basal layer, move upward, and execute a terminal differentiation program consisting of three distinct stages: spinous, granular layer, and stratum corneum. Notch signaling has been implicated in late stages of differentiation, but the commitment switch remains unknown. Here we show with loss and gain-of function studies that active Notch intracellular domain (NICD) and its obligate canonical signaling partner RBP-J act at the basal/suprabasal juncture to induce spinous and down-regulate basal fate. Spinous layers are absent in RBP-J conditional null epidermis and expanded when Notch1 signaling is elevated transgenically in epidermis. We show that RBP-J is essential for mediating both spinous gene activation and basal gene repression. In contrast, the NICD/RBP-J target gene Hes1 is expressed in spinous layers and mediates spinous gene induction but not basal gene repression. These data uncover an early role for RBP J and Notch in commitment of epidermal cells to terminally differentiate and reveal that spinous gene induction is mediated by a Hes1-dependent mechanism, while basal gene repression occurs independently of Hes1. PMID- 17079690 TI - Hedgehog signaling and the retina: insights into the mechanisms controlling the proliferative properties of neural precursors. AB - Hedgehog signaling has been linked to cell proliferation in a variety of systems; however, its effects on the cell cycle have not been closely studied. In the vertebrate retina, Hedgehog's effects are controversial, with some reports emphasizing increased proliferation and others pointing to a role in cell cycle exit. Here we demonstrate a novel role for Hedgehog signaling in speeding up the cell cycle in the developing retina by reducing the length of G1 and G2 phases. These fast cycling cells tend to exit the cell cycle early. Conversely, retinal progenitors with blocked Hedgehog signaling cycle more slowly, with longer G1 and G2 phases, and remain in the cell cycle longer. Hedgehog may modulate cell cycle kinetics through activation of the key cell cycle activators cyclin D1, cyclin A2, cyclin B1, and cdc25C. These findings support a role for Hedgehog in regulating the conversion from slow cycling stem cells to fast cycling transient amplifying progenitors that are closer to cell cycle exit. PMID- 17079691 TI - Personal hair dye use and risks of glioma, meningioma, and acoustic neuroma among adults. AB - Previous studies have suggested an association of personal hair dye use with bladder and hematopoietic cancers. Risks for brain tumors are not well understood. The authors investigated associations between use of synthetic hair dyes and risk of brain tumors in a hospital-based case-control study. The study included adults newly diagnosed with glioma (n = 489), meningioma (n = 197), or acoustic neuroma (n = 96) between 1994 and 1998 at three urban US hospitals and 799 controls. Odds ratios were estimated and 95% confidence intervals were calculated using unconditional logistic regression. Detailed exposure histories were obtained by interview. There was no consistent pattern of elevated odds ratios for glioma, meningioma, or acoustic neuroma with use or prolonged use of permanent, semipermanent, temporary, or gradual hair dyes. Although use of permanent brown hair dye for 20 or more years was associated with glioma among women, the estimate was imprecise (odds ratio = 3.8, 95% confidence interval: 1.2, 12.5) and was based on just 13 exposed cases; thus, this could be a chance finding. Overall, there was little consistent evidence for an association of synthetic hair dye use with glioma, meningioma, or acoustic neuroma. However, prolonged use of dark-colored permanent dyes warrants further investigation given the high prevalence of hair dyeing. PMID- 17079692 TI - Bladder cancer and exposure to water disinfection by-products through ingestion, bathing, showering, and swimming in pools. AB - Bladder cancer has been associated with exposure to chlorination by-products in drinking water, and experimental evidence suggests that exposure also occurs through inhalation and dermal absorption. The authors examined whether bladder cancer risk was associated with exposure to trihalomethanes (THMs) through ingestion of water and through inhalation and dermal absorption during showering, bathing, and swimming in pools. Lifetime personal information on water consumption and water-related habits was collected for 1,219 cases and 1,271 controls in a 1998-2001 case-control study in Spain and was linked with THM levels in geographic study areas. Long-term THM exposure was associated with a twofold bladder cancer risk, with an odds ratio of 2.10 (95% confidence interval: 1.09, 4.02) for average household THM levels of >49 versus < or =8 micro g/liter. Compared with subjects not drinking chlorinated water, subjects with THM exposure of >35 micro g/day through ingestion had an odds ratio of 1.35 (95% confidence interval: 0.92, 1.99). The odds ratio for duration of shower or bath weighted by residential THM level was 1.83 (95% confidence interval: 1.17, 2.87) for the highest compared with the lowest quartile. Swimming in pools was associated with an odds ratio of 1.57 (95% confidence interval: 1.18, 2.09). Bladder cancer risk was associated with long-term exposure to THMs in chlorinated water at levels regularly occurring in industrialized countries. PMID- 17079693 TI - Phase II trial of capecitabine and oxaliplatin (CAPOX) plus cetuximab in patients with metastatic colorectal cancer who progressed after oxaliplatin-based chemotherapy. AB - BACKGROUND: Cetuximab is an IgG1 monoclonal antibody targeting the epidermal growth factor receptor and is able to reverse the resistance to irinotecan in patients with metastatic colorectal cancer (mCRC). This phase II trial evaluates the safety and efficacy of cetuximab combined with capecitabine and oxaliplatin (CAPOX) in the treatment of patients with mCRC progressing under oxaliplatin based chemotherapy. PATIENTS AND TREATMENT: Forty patients with mCRC were treated with cetuximab (loading dose 400 mg/m(2) and then 250 mg/m(2) i.v. weekly) in combination with CAPOX (d(1): L-OHP 85 mg/m(2) and d(1-7) capecitabine 2000 mg/m(2) every 2 weeks). Thirty-one (77.5%) and nine (22.5%) patients had oxaliplatin-refractory and -resistant disease, respectively; in addition, 32 (80%) patients had also progressed on prior irinotecan-based chemotherapy. RESULTS: One hundred and thirty-four cycles were administered (median of four cycles per patient). Main toxic effects included grade 3-4 neutropenia (12.5%), grade 3/4 diarrhea (7.5%), grade 3 fatigue (2.5%), and grade 2-3 neurotoxicity (22.5%). One (2.5%) complete and seven (17.5%) partial responses were achieved [overall objective response rate (ORR): 20%; 95% confidence interval (CI): 9% to 32%)], whereas 11 (27.5%) patients had stable disease [disease control rate (DCR): 47.5%; 95% CI: 30.2% to 64.5%]. The ORR and DCR were 18.7% and 46.8%, respectively, in patients with oxaliplatin-refractory disease. The median time to tumor progression was 3 months, the median survival 10.7 months and the probability of 1-year survival rate 53.4%. CONCLUSIONS: The combination of cetuximab plus CAPOX is safe and has a promising activity in patients with mCRC refractory or resistant to oxaliplatin. PMID- 17079694 TI - Randomized phase III study of cisplatin plus irinotecan versus carboplatin plus paclitaxel, cisplatin plus gemcitabine, and cisplatin plus vinorelbine for advanced non-small-cell lung cancer: Four-Arm Cooperative Study in Japan. AB - BACKGROUND: To compare the efficacy and toxicity of three platinum-based combination regimens against cisplatin plus irinotecan (IP) in patients with untreated advanced non-small-cell lung cancer (NSCLC) by a non-inferiority design. PATIENTS AND METHODS: A total of 602 patients were randomly assigned to one of four regimens: cisplatin 80 mg/m(2) on day 1 plus irinotecan 60 mg/m(2) on days 1, 8, 15 every 4 weeks (IP) carboplatin AUC 6.0 min x mg/mL (area under the concentration-time curve) on day 1 plus paclitaxel 200 mg/m(2) on day 1 every 3 weeks (TC); cisplatin 80 mg/m(2) on day 1 plus gemcitabine 1000 mg/m(2) on days 1, 8 every 3 weeks (GP); and cisplatin 80 mg/m(2) on day 1 plus vinorelbine 25 mg/m(2) on days 1, 8 every 3 weeks (NP). RESULTS: The response rate, median survival time, and 1-year survival rate were 31.0%, 13.9 months, 59.2%, respectively, in IP; 32.4%, 12.3 months, 51.0% in TC; 30.1%, 14.0 months, 59.6% in GP; and 33.1%, 11.4 months, 48.3% in NP. No statistically significant differences were found in response rate or overall survival, but the non inferiority of none of the experimental regimens could be confirmed. All the four regimens were well tolerated. CONCLUSION: The four regimens have similar efficacy and different toxicity profiles, and they can be used to treat advanced NSCLC patients. PMID- 17079695 TI - A high incidence of late-onset neutropenia following rituximab-containing chemotherapy as a primary treatment of CD20-positive B-cell lymphoma: a single institution study. AB - BACKGROUND: Late-onset neutropenia (LON) has been reported following rituximab containing chemotherapy. Its incidence and risk factors, however, have not been extensively studied. PATIENTS AND METHODS: We retrospectively reviewed the medical records of 107 patients treated with rituximab-containing chemotherapy as a primary treatment of CD20-positive B-cell lymphomas and identified cases with LON as defined by the neutrophil count of 20 million per mL), data adjusted for age and period of abstinence showed a decreasing trend for sperm density over time, R2=0.45 (P=.017). There was no such trend in sperm motility and motile density. However, this trend has to be interpreted with caution due to fluctuations in semen parameters, population bias and the retrospective nature of the analysis. PMID- 17079744 TI - Sperm characteristics and DNA integrity of Iberian red deer (Cervus elaphus hispanicus) epididymal spermatozoa frozen in the presence of enzymatic and nonenzymatic antioxidants. AB - The main goal of this study was to investigate the potential protective effects of enzymatic and nonenzymatic antioxidants on cryopreservation injuries to red deer epididymal spermatozoa. In Experiment 1, the effects on sperm freezability of the enzymatic antioxidants catalase, superoxide dismutase, and a combination thereof were studied. In Experiment 2, sperm cryoresistance was evaluated when different nonenzymatic antioxidants, such as vitamin E, vitamin C, and butylated hydroxytoluene (BHT), were added to the freezing extender. Sperm quality was judged in vitro by microscopic assessments of individual sperm motility (SMI), viability, and acrosome (ie, spermatozoa with normal apical ridges; % NAR) and membrane (by means of the HOS test) integrity. To address fully these topics, we incorporated a new set of functional sperm tests for mitochondrial function, membrane phospholipid disorder, and sperm chromatin stability. Samples were evaluated after freezing and thawing, and after a 2-hour period of incubation at 37 degrees C. The present study demonstrates that the addition of enzymatic antioxidants to freezing extenders improves sperm viability after cooling, and improves sperm motility, acrosome integrity, and mitochondrial status (P<.05) after thawing. After a 2-hour incubation period at 37 degrees C in the presence of enzymatic antioxidants, an improvement in membrane integrity (P<.05) was observed. However, when nonenzymatic antioxidants were present in the freezing diluents, no positive effects on thawed sperm parameters were noted. The chromatin stability test did not show significant differences between the treatments. We conclude that enzymatic antioxidants should be present in the early steps of cryopreservation of epididymal spermatozoa from red deer, so as to improve motility and acrosome integrity. PMID- 17079745 TI - Management of post-ejaculatory perineal pain. PMID- 17079746 TI - Metabolic costs and muscle activity patterns during robotic- and therapist assisted treadmill walking in individuals with incomplete spinal cord injury. AB - BACKGROUND AND PURPOSE: Robotic devices that provide passive guidance and stabilization of the legs and trunk during treadmill stepping may increase the delivery of locomotor training to subjects with neurological injury. Lower extremity guidance also may reduce voluntary muscle activity as compared with compliant assistance provided by therapists. The purpose of this study was to investigate differences in metabolic costs and lower-limb muscle activity patterns during robotic- and therapist-assisted treadmill walking. SUBJECTS: Twelve ambulatory subjects with motor incomplete spinal cord injury participated. METHODS: In 2 separate protocols, metabolic and electromyographic (EMG) data were collected during standing and stepping on a treadmill with therapist and robotic assistance. During robotic-assisted walking, subjects were asked to match the kinematic trajectories of the device and maximize their effort. During therapist assisted walking, subjects walked on the treadmill with manual assistance provided as necessary. RESULTS: Metabolic costs and swing-phase hip flexor EMG activity were significantly lower when subjects were asked to match the robotic device trajectories than with therapist-assisted walking. These differences were reduced when subjects were asked to maximize their effort during robotic-assisted stepping, although swing-phase plantar-flexor EMG activity was increased. In addition, during standing prior to therapist- or robotic-assisted stepping, metabolic costs were higher without stabilization from the robotic device. DISCUSSION AND CONCLUSION: Differences in metabolic costs and muscle activity patterns between therapist- and robotic-assisted standing and stepping illustrate the importance of minimizing passive guidance and stabilization provided during step training protocols. PMID- 17079747 TI - A comparison of physical functional performance and strength in women with fibromyalgia, age- and weight-matched controls, and older women who are healthy. AB - BACKGROUND AND PURPOSE: The purpose of this study was to compare functionality and strength among women with fibromyalgia (FM), women without FM, and older women. SUBJECTS: Twenty-nine women with FM (age [X+/-SD]=46+/-7 years), 12 age- and weight-matched women without FM (age=44+/-8 years), and 38 older women who were healthy (age=71+/-7 years) participated. METHODS: The Continuous Scale Physical Functional Performance Test (CS-PFP) was used to assess functionality. Isokinetic leg strength was measured at 60 degrees/s, and handgrip strength was measured using a handgrip dynamometer. RESULTS: The women without FM had significantly higher functionality scores compared with women with FM and older women. There were no differences in functionality between women with FM and older women. Strength measures for the leg were higher in women without FM compared with women with FM and older women, and both women with and without FM had higher grip strengths compared with older women. DISCUSSION AND CONCLUSION: This study demonstrated that women with FM and older women who are healthy have similar lower-body strength and functionality, potentially enhancing the risk for premature age-associated disability. PMID- 17079748 TI - Performance measures provide assessments of pain and function in people with advanced osteoarthritis of the hip or knee. AB - BACKGROUND AND PURPOSE: Pain and physical function are core outcome measures for people with osteoarthritis, and self-report questionnaires have been the preferred assessment method. There is evidence suggesting that self-reports of physical function represent what people experience when performing activities rather than their ability to perform activities. The purpose of this study was to examine the factorial validity of performance-specific assessments of pain and function. SUBJECTS: The sample consisted of 177 participants who had osteoarthritis of the hip (n=81) or knee (n=96) and who were awaiting total joint arthroplasty. METHODS: Through a cross-sectional design, participants performed 4 performance activities (self-paced walk test, stair test, Timed "Up & Go" Test, and Six-Minute Walk Test). OUTCOMES: were time or distance (function) and pain ratings obtained immediately after each activity. The authors conceptualized 2 correlated factors, with pain items loading uniquely on 1 factor and functional items loading on the second factor, and uncorrelated error terms. Confirmatory factor analysis was applied. RESULTS: Initial analysis yielded results consistent with the conceptualized model in this study with the exception of a nonzero correlation between the stair pain and function error terms. Dropping the stair test provided results consistent with the conceptualized model. DISCUSSION AND CONCLUSION: Given the limitations of self-report alone as a method of obtaining reasonably distinct assessments of pain and function, the extent to which performance-specific assessments could accomplish this goal was examined in this study. It was found that collectively the walk test, Timed "Up & Go" Test, and Six-Minute Walk Test yielded 2 factors consistent with the health concepts of pain and function. The authors believe that the application of these tests may provide clinicians and clinical researchers with more distinct impressions of pain and function that complement information from self-report measures. PMID- 17079749 TI - Invited commentary. PMID- 17079750 TI - Gait characteristics of elderly people with a history of falls: a dynamic approach. AB - BACKGROUND AND PURPOSE: This study investigated changes in the kinematics of elderly people who experienced at least one fall 6 months prior to data collection. The authors hypothesized that, in order to decrease variability of walking, people with a history of falls would show different kinematic adaptations of their walking patterns compared with elderly people with no history of falls. SUBJECTS AND METHODS: Twenty-one elderly people who had fallen within the previous 6 months ("fallers"; mean age=72.1 years, SD=4.9) and 27 elderly people with no history of falls ("nonfallers"; mean age=73.8 years, SD=6.4) walked at their preferred stride frequency (STF) as treadmill speed was gradually increased (from 0.18 m/s to 1.52 m/s) and then decreased in steps of 0.2 m/s. Gait parameter measurements were recorded, and statistical analysis was applied using walking speed and STF as independent variables. RESULTS: Fifty seven percent of the fallers were unable to walk at the fastest speed, whereas all nonfallers walked comfortably at all walking speeds. Although the fallers showed significantly greater STF, smaller stride lengths, smaller center-of-mass lateral sway, and smaller ankle plantar flexion and hip extension during push off, they showed increased variability of kinematic measures in their coordination of walking compared with the nonfallers. DISCUSSION AND CONCLUSION: Although the fallers' adaptations were expected to reduce variability in the coordination of walking, they showed less stable gait patterns (ie, greater variability) compared with the nonfallers. Increased variability of walking patterns may be an important gait risk factor in elderly people with a history of falls. PMID- 17079751 TI - Managing a patient's constipation with physical therapy. AB - BACKGROUND AND PURPOSE: Constipation is a prevalent condition in the United States, with typical treatment consisting of diet modification, stool softeners, and laxatives. These interventions, however, are not always effective. The purpose of this case report is to describe the use of abdominal massage in physical therapist management for a patient with constipation. CASE DESCRIPTION: An 85-year-old woman with constipation was referred for physical therapy following unsuccessful treatment with stool softeners. The patient was instructed in bowel management as well as a daily, 10-minute home abdominal massage program. OUTCOMES: Upon re-examination, the patient reported a return of normal bowel frequency and function without the need to strain or use digital evacuation. DISCUSSION: Physical therapy incorporating abdominal massage appeared to be helpful in resolving this patient's constipation. Unlike medical management of constipation, no known side effects have been identified with abdominal massage. PMID- 17079752 TI - Lessons learned in participant recruitment and retention: the EXCITE trial. AB - Participant recruitment is considered the most difficult aspect of the research process. Despite the integral role of recruitment in randomized clinical trials, publication of data defining the recruitment effort is not routine in rehabilitation initiatives. The recruitment process for the Extremity Constraint Induced Therapy Evaluation (EXCITE) trial illustrates obstacles to and strategies for participant accrual and retention that are inherent in rehabilitation clinical trials. The purpose of this article is to increase awareness of the multiple facets of recruitment necessary for successful clinical trials, thus supporting the continued development of evidence-based practice in physical therapy. The Recruitment Index is presented as a variable to measure recruitment efficacy. In addition, ethical aspects of recruitment are explored, including informed consent and the concept of therapeutic misconception. PMID- 17079753 TI - In the best interests of the patient. PMID- 17079754 TI - Leaving a legacy. PMID- 17079755 TI - Does electrical stimulation improve motor recovery in patients with idiopathic facial (Bell) palsy? PMID- 17079756 TI - Images in clinical medicine. Necrobiosis lipoidica diabeticorum. PMID- 17079757 TI - Paying for performance--risks and recommendations. PMID- 17079758 TI - Public report cards--cardiac surgery and beyond. PMID- 17079759 TI - Epirubicin and cyclophosphamide, methotrexate, and fluorouracil as adjuvant therapy for early breast cancer. AB - BACKGROUND: The National Epirubicin Adjuvant Trial (NEAT) and the BR9601 trial examined the efficacy of anthracyclines in the adjuvant treatment of early breast cancer. METHODS: In NEAT, we compared four cycles of epirubicin followed by four cycles of cyclophosphamide, methotrexate, and fluorouracil (CMF) with six cycles of CMF alone. In the BR9601 trial, we compared four cycles of epirubicin followed by four cycles of CMF, with eight cycles of CMF alone every 3 weeks. The primary end points were relapse-free and overall survival. The secondary end points were adverse effects, dose intensity, and quality of life. RESULTS: The two trials included 2391 women with early breast cancer; the median follow-up was 48 months. Relapse-free and overall survival rates were significantly higher in the epirubicin-CMF groups than in the CMF-alone groups (2-year relapse-free survival, 91% vs. 85%; 5-year relapse-free survival, 76% vs. 69%; 2-year overall survival, 95% vs. 92%; 5-year overall survival, 82% vs. 75%; P<0.001 by the log-rank test for all comparisons). Hazard ratios for relapse (or death without relapse) (0.69; 95% confidence interval [CI], 0.58 to 0.82; P<0.001) and death from any cause (0.67; 95% CI, 0.55 to 0.82; P<0.001) favored epirubicin plus CMF over CMF alone. Independent prognostic factors were nodal status, tumor grade, tumor size, and estrogen-receptor status (P<0.001 for all four factors) and the presence or absence of vascular or lymphatic invasion (P=0.01). These factors did not significantly interact with the effect of epirubicin plus CMF. The overall incidence of adverse effects was significantly higher with epirubicin plus CMF than with CMF alone but did not significantly affect the delivered-dose intensity or the quality of life. CONCLUSIONS: Epirubicin plus CMF is superior to CMF alone as adjuvant treatment for early breast cancer. (ClinicalTrials.gov number, NCT00003577 [ClinicalTrials.gov].). PMID- 17079760 TI - Colonoscopy in colorectal-cancer screening for detection of advanced neoplasia. AB - BACKGROUND: Recommendations for colorectal-cancer screening are based solely on age and family history of cancer, not sex. METHODS: We performed a cross sectional analysis of the data from a large colonoscopy-based screening program that included 50,148 participants who were 40 to 66 years of age. People 40 to 49 years of age were eligible only if they had a family history of cancer of any type. Of the 43,042 participants 50 to 66 years of age, 13.3% reported a family history of colorectal cancer, as did 66.3% of the 7106 participants who were 40 to 49 years of age. We defined advanced neoplasia as cancer or adenoma that was at least 10 mm in diameter, had high-grade dysplasia, or had villous or tubulovillous histologic characteristics, or any combination thereof. We used multivariate logistic regression to identify associations between participants' characteristics and advanced neoplasia in a primary (or derivation) data set, and we confirmed the associations in a secondary (or validation) data set. RESULTS: Advanced neoplasia was detected in 2553 (5.9%) participants 50 to 66 years of age and in 243 (3.4%) participants 40 to 49 years of age. The rate of complications during colonoscopy was 0.1%, and no participants died. In the validation set, a logistic-regression model showed that male sex was independently associated with advanced neoplasia (adjusted odds ratio, 1.73; 95% confidence interval, 1.52 to 1.98; P<0.001). In each age group (40 to 49 years, 50 to 54 years, 55 to 59 years, and 60 to 66 years), the number of persons who would have to undergo colorectal-cancer screening in order to detect one advanced neoplasia was significantly lower in men than in women (23 vs. 36, 17 vs. 28, 12 vs. 22, and 10 vs. 18, respectively). CONCLUSIONS: We detected advanced neoplasia at a significantly higher rate in men than in women, which may warrant refinement of the screening recommendations for colorectal cancer. PMID- 17079761 TI - Left ventricular assist device and drug therapy for the reversal of heart failure. AB - BACKGROUND: In patients with severe heart failure, prolonged unloading of the myocardium with the use of a left ventricular assist device has been reported to lead to myocardial recovery in small numbers of patients for varying periods of time. Increasing the frequency and durability of myocardial recovery could reduce or postpone the need for subsequent heart transplantation. METHODS: We enrolled 15 patients with severe heart failure due to nonischemic cardiomyopathy and with no histologic evidence of active myocarditis. All had markedly reduced cardiac output and were receiving inotropes. The patients underwent implantation of left ventricular assist devices and were treated with lisinopril, carvedilol, spironolactone, and losartan to enhance reverse remodeling. Once regression of left ventricular enlargement had been achieved, the beta2-adrenergic-receptor agonist clenbuterol was administered to prevent myocardial atrophy. RESULTS: Eleven of the 15 patients had sufficient myocardial recovery to undergo explantation of the left ventricular assist device a mean (+/-SD) of 320+/-186 days after implantation of the device. One patient died of intractable arrhythmias 24 hours after explantation; another died of carcinoma of the lung 27 months after explantation. The cumulative rate of freedom from recurrent heart failure among the surviving patients was 100% and 88.9% 1 and 4 years after explantation, respectively. The quality of life as assessed by the Minnesota Living with Heart Failure Questionnaire score at 3 years was nearly normal. Fifty nine months after explantation, the mean left ventricular ejection fraction was 64+/-12%, the mean left ventricular end-diastolic diameter was 59.4+/-12.1 mm, the mean left ventricular end-systolic diameter was 42.5+/-13.2 mm, and the mean maximal oxygen uptake with exercise was 26.3+/-6.0 ml per kilogram of body weight per minute. CONCLUSIONS: In this single-center study, we found that sustained reversal of severe heart failure secondary to nonischemic cardiomyopathy could be achieved in selected patients with the use of a left ventricular assist device and a specific pharmacologic regimen. PMID- 17079762 TI - Treatment of periodontal disease and the risk of preterm birth. AB - BACKGROUND: Maternal periodontal disease has been associated with an increased risk of preterm birth and low birth weight. We studied the effect of nonsurgical periodontal treatment on preterm birth. METHODS: We randomly assigned women between 13 and 17 weeks of gestation to undergo scaling and root planing either before 21 weeks (413 patients in the treatment group) or after delivery (410 patients in the control group). Patients in the treatment group also underwent monthly tooth polishing and received instruction in oral hygiene. The gestational age at the end of pregnancy was the prespecified primary outcome. Secondary outcomes were birth weight and the proportion of infants who were small for gestational age. RESULTS: In the follow-up analysis, preterm birth (before 37 weeks of gestation) occurred in 49 of 407 women (12.0%) in the treatment group (resulting in 44 live births) and in 52 of 405 women (12.8%) in the control group (resulting in 38 live births). Although periodontal treatment improved periodontitis measures (P<0.001), it did not significantly alter the risk of preterm delivery (P=0.70; hazard ratio for treatment group vs. control group, 0.93; 95% confidence interval [CI], 0.63 to 1.37). There were no significant differences between the treatment and control groups in birth weight (3239 g vs. 3258 g, P=0.64) or in the rate of delivery of infants that were small for gestational age (12.7% vs. 12.3%; odds ratio, 1.04; 95% CI, 0.68 to 1.58). There were 5 spontaneous abortions or stillbirths in the treatment group, as compared with 14 in the control group (P=0.08). CONCLUSIONS: Treatment of periodontitis in pregnant women improves periodontal disease and is safe but does not significantly alter rates of preterm birth, low birth weight, or fetal growth restriction. (ClinicalTrials.gov number, NCT00066131 [ClinicalTrials.gov].). PMID- 17079763 TI - Pay for performance in commercial HMOs. AB - BACKGROUND: Pay for performance has increasingly become the subject of intense interest and debate, both of which have been heightened as the Centers for Medicare and Medicaid Services moves closer to adopting this approach for Medicare. Although many claims have been made for the effectiveness of this approach, the extent of its national penetration remains unknown. METHODS: We surveyed a sample of 252 health maintenance organizations (HMOs) (response rate, 96%) drawn from 41 metropolitan areas across the nation about use of pay for performance. We determined the prevalence of pay-for-performance programs, detailed the features of such programs, and examined the adoption of pay for performance as a function of the characteristics of both the health plans and markets. RESULTS: More than half the HMOs, representing more than 80% of persons enrolled, use pay for performance in their provider contracts. Of the 126 health plans with pay-for-performance programs, nearly 90% had programs for physicians and 38% had programs for hospitals. Use of pay for performance was statistically associated with geographic region, use of primary care providers (PCPs) as gatekeepers, use of capitation to pay PCPs, and whether the plans themselves received bonuses or penalties according to performance. CONCLUSIONS: Pay for performance is now commonly used by HMOs, especially those that are situated to assign responsibility for a particular patient to a PCP or medical group. As the design of Medicare with pay for performance moves forward, it will be important to leverage the early experience of pay for performance in the commercial market. PMID- 17079764 TI - Clinical practice. Management of hyperglycemia in the hospital setting. PMID- 17079765 TI - Images in clinical medicine. Progression of a pedunculated polyp. PMID- 17079766 TI - Clinical problem-solving. The missing piece. PMID- 17079767 TI - Adjuvant chemotherapy for breast cancer--30 years later. PMID- 17079768 TI - When the failing, end-stage heart is not end-stage. PMID- 17079769 TI - Preterm birth and periodontal disease. PMID- 17079770 TI - Retraction: Sudbo J et al. DNA content as a prognostic marker in patients with oral leukoplakia. N Engl J Med 2001;344:1270-8 and Sudbo J et al. The influence of resection and aneuploidy on mortality in oral leukoplakia. N Engl J Med 2004;350:1405-13. PMID- 17079771 TI - Voters and health care in the 2006 election. PMID- 17079772 TI - Resistant or difficult-to-control hypertension. PMID- 17079773 TI - The severe gout of Emperor Charles V. PMID- 17079774 TI - Dystonia. PMID- 17079775 TI - Case 24-2006: a woman with hypotension after an overdose of amlodipine. PMID- 17079776 TI - Tracheal replacement with aortic allografts. PMID- 17079777 TI - Diversity of the angiogenic phenotype in non-small cell lung cancer. AB - Angiogenesis is crucial for tumor biology. There are many mechanisms by which tumors induce angiogenesis. We hypothesize that each individual tumor develops a unique mechanism to induce angiogenesis, and that activation of a particular angiogenic pathway suppresses the evolution of alternative pathways. We characterized 168 human non-small cell lung cancer (NSCLC) specimens for levels of angiogenic factors (angiogenic CXC chemokines, basic fibroblast growth factor, and vascular endothelial growth factor). We also induced lung tumor formation in A/J mice by injecting the tobacco carcinogen NNK. We dissected individual lung tumors and measured expression of angiogenic factors from three distinct families using real-time PCR. Finally, we controlled the angiogenic milieu using in vivo models to determine the resultant phenotype of the angiogenic factors expressed by NSCLC cells. Human tumors displayed marked variation in the expression of angiogenic factors. Individual mouse tumors, even from within the same mouse, displayed variability in their pattern of expression of angiogenic factors. In a sponge model of angiogenesis using murine lung cancer cells, implanting LLC cells with an angiogenic factor suppressed the expression of other angiogenic factors in implanted sponges. This suppressive effect was not seen in vitro. We conclude that lung cancer tumors evolve a unique and dominant angiogenic phenotype. Once an angiogenic pathway is activated, it may allow for tumor growth to proceed in the absence of a selection pressure to activate a second pathway. PMID- 17079778 TI - Effect of concentrated ambient particles on macrophage phagocytosis and killing of Streptococcus pneumoniae. AB - Particulate air pollution is linked to increased pneumonia epidemiologically and diminished lung bacterial clearance experimentally. We investigated the effect of concentrated ambient particles (CAPs, or =15, drop in hematocrit of > or = 10 points, or the patient requiring a blood transfusion. Patients were divided into a low-volume (NS < 2L) and a high-volume (NS 2L) group. Data were reported as mean (+/-SD). Student's t- and Wilcoxon tests were used to compare data. Receiver operating characteristic (ROC) curves tested the utility of BD-4 in differentiating minor from major injury in the study groups. RESULTS: Four hundred eighty-nine trauma patients (mean age, 36 [+/-18] yr) were enrolled; 82% were male, and 34% had penetrating injury. Major-(20%) compared with minor-(80%) injury patients were significantly (p = 0.0001) more acidotic (BD-0 mean difference: -3.3 mmol/L; 95% confidence interval [CI] = -2.5 to -4.2). The high-volume group (n = 174) received 3,342 (+/-1,821) mL, and the low-volume group (n = 315) received 621 (+/-509) mL of NS. Areas under the ROC curves for the high-volume (0.63; 95% CI = 0.52 to 0.74) and low-volume (0.73; 95% CI = 0.60 to 0.86) groups were not significantly different from each other. CONCLUSIONS: Base deficit was able to distinguish minor from major injury after four hours of resuscitation, irrespective of the volume of NS infused. PMID- 17079787 TI - Effect of point-of-care influenza testing on management of febrile children. AB - OBJECTIVES: To determine the effect of point-of-care testing (POCT) for influenza on the physician management of febrile children who are at risk for serious bacterial illness (SBI) on the basis of age and temperature and who are presenting to a pediatric emergency department (ED) during an influenza outbreak. METHODS: Patients 2-3 months of age with temperature of > or = 38 degrees C and patients 3-24 months of age with temperature of > or = 39 degrees C who were presenting to a pediatric ED during an influenza outbreak were enrolled into a prospective, quasi-randomized, controlled trial. Influenza testing was performed on enrolled patients by either the POCT or the standard-testing (ST) methods. The two groups were compared in terms of laboratory testing, chest radiography, antibiotic use, visit-associated costs, pediatric ED lengths of stay, inpatient admission, and return visits to the pediatric ED. Similar analyses also were performed on the resulting subgroups of patients on the basis of method of testing (POCT or ST) and test result (positive or negative). RESULTS: Of 767 eligible patients, 700 (91%) completed the study. No significant differences were demonstrated between the POCT and ST groups with respect to laboratory tests ordered, chest radiographs obtained, antibiotic administration, inpatient admission, return visits to the pediatric ED, lengths of stay, or visit associated costs. In the subgroup analysis, the adjusted odds ratios (ORs) for blood culture in influenza test-positive to -negative patients were 0.59 and 0.71 in the POCT and ST groups, respectively (p = 0.088). The adjusted ORs for urine culture in influenza test-positive to -negative patients were 0.46 and 0.67 in the POCT and ST groups, respectively (p = 0.005). CONCLUSIONS: When using a strategy of performing influenza testing on all patients at risk for SBI who presented to a pediatric ED during an influenza outbreak, the method of testing (POCT or ST) did not appear to significantly alter physician management, cost, or length of stay in the pediatric ED. However, if the interaction of the method of testing and the test result (positive or negative) were considered, a positive POCT for influenza was associated with a significant reduction in orders for urinalyses and urine cultures. PMID- 17079788 TI - Stylet bend angles and tracheal tube passage using a straight-to-cuff shape. AB - OBJECTIVES: Malleable stylets improve maneuverability and control during tube insertion, but after passage through the vocal cords the stiffened tracheal tube may impinge on the tracheal rings, preventing passage. The goal of this study was to assess insertion difficulty with styletted tubes of different bend angles. METHODS: Tube passage was assessed with four different bend angles (25 degrees, 35 degrees, 45 degrees, and 60 degrees) using straight-to-cuff-shaped tubes. In two separate airway procedure classes, 16 operators in each class (32 total) placed randomly ordered styletted tubes of the different angles into eight cadavers (16 total). Operators subjectively graded the ease of tube passage as no resistance, some resistance, or impossible to advance. RESULTS: No resistance was reported in 69.1% (177/256) at 25 degrees, in 63.7% (163/256) at 35 degrees, in 39.4% (101/256) at 45 degrees, and in 8.9% (22/256) at 60 degrees. Tube passage was impossible in 2.3% of insertions (6/256) at 25 degrees, in 3.5% (9/256) at 35 degrees, in 11.3% (29/256) at 45 degrees, and in 53.9% (138/256) at 60 degrees. The odds ratios of impossible tube passage for 35 degrees, 45 degrees, and 60 degrees vs. 25 degrees were 1.52 (95% confidence interval [CI] = 0.55 to 4.16), 5.32 (95% CI = 2.22 to 12.71), and 48.72 (95% CI = 21.35 to 111.03), respectively. CONCLUSIONS: Bend angles beyond 35 degrees with straight-to-cuff styletted tracheal tubes increase the risk of difficult and impossible tube passage into the trachea. The authors did not compare different stylet stopping points, stylets of different stiffness, or tracheal tubes with different tip designs, all variables that can affect tube passage. PMID- 17079789 TI - A prospective comparison of ultrasound-guided and blindly placed radial arterial catheters. AB - BACKGROUND: Arterial cannulation for continuous blood-pressure measurement and frequent arterial-blood sampling commonly are required in critically ill patients. OBJECTIVES: To compare ultrasound (US)-guided versus traditional palpation placement of arterial lines for time to placement, number of attempts, sites used, and complications. METHODS: This was a prospective, randomized interventional study at a Level 1 academic urban emergency department with an annual census of 78,000 patients. Patients were randomized to either palpation or US-guided groups. Inclusion criteria were any adult patient who required an arterial line according to the treating attending. Patients who had previous attempts at an arterial line during the visit, or who could not be randomized because of time constraints, were excluded. Enrollment was on a convenience basis, during hours worked by researchers over a six-month period. Patients in either group who had three failed attempts were rescued with the other technique for patient comfort. Statistical analysis included Fisher's exact, Mann-Whitney, and Student's t-tests. RESULTS: Sixty patients were enrolled, with 30 patients randomized to each group. Patients randomized to the US group had a shorter time required for arterial line placement (107 vs. 314 seconds; difference, 207 seconds; p = 0.0004), fewer placement attempts (1.2 vs. 2.2; difference, 1; p = 0.001), and fewer sites required for successful line placement (1.1 vs. 1.6; difference, 0.5; p = 0.001), as compared with the palpation group. CONCLUSIONS: In this study, US guidance for arterial cannulation was successful more frequently and it took less time to establish the arterial line as compared with the palpation method. PMID- 17079790 TI - Unified medical language system coverage of emergency-medicine chief complaints. AB - BACKGROUND: Emergency department (ED) chief-complaint (CC) data increasingly are important for clinical-care and secondary uses such as syndromic surveillance. There is no widely used ED CC vocabulary, but experts have suggested evaluation of existing health-care vocabularies for ED CC. OBJECTIVES: To evaluate the ED CC coverage in existing biomedical vocabularies from the Unified Medical Language System (UMLS). METHODS: The study sample included all CC entries for all visits to three EDs over one year. The authors used a special-purpose text processor to clean CC entries, which then were mapped to UMLS concepts. The UMLS match rates then were calculated and analyzed for matching concepts and nonmatching entries. RESULTS: A total of 203,509 ED visits was included. After cleaning with the text processor, 82% of the CCs matched a UMLS concept. The authors identified 5,617 unique UMLS concepts in the ED CC data, but many were used for only one or two visits. One thousand one hundred thirty-six CC concepts were used more than ten times and covered 99% of all the ED visits. The largest biomedical vocabulary in the UMLS is the Systematized Nomenclature of Medicine Clinical Terms (SNOMED CT), which included concepts for 79% of all ED CC entries. However, some common CCs were not found in SNOMED CT. CONCLUSIONS: The authors found that ED CC concepts are well covered by the UMLS and that the best source of vocabulary coverage is from SNOMED CT. There are some gaps in UMLS and SNOMED CT coverage of ED CCs. Future work on vocabulary control for ED CCs should build upon existing vocabularies. PMID- 17079791 TI - Integrating the core competencies: proceedings from the 2005 Academic Assembly consortium. AB - The Accreditation Council for Graduate Medical Education mandated the integration of the core competencies into residency training in 2001. To this end, educators in emergency medicine (EM) have been proactive in their approach, using collaborative efforts to develop methods that teach and assess the competencies. The first steps toward a collaborative approach occurred during the proceedings of the Council of Emergency Medicine Residency Directors (CORD-EM) academic assembly in 2002. Three years later, the competencies were revisited by working groups of EM program directors and educators at the 2005 Academic Assembly. This report provides a summary discussion of the status of integration of the competencies into EM training programs in 2005. PMID- 17079792 TI - Increased lipid rafts and accelerated lipopolysaccharide-induced tumor necrosis factor-alpha secretion in Abca1-deficient macrophages. AB - Lipid rafts on the cell surface are believed to be very important as platforms for various cellular functions. The aim of this study was to know whether defective lipid efflux may influence lipid rafts on the cell surface and their related cellular functions. We investigated macrophages with defective lipid efflux from ATP binding cassette transporter A1-deficient (Abca1-KO) mice. Lipid rafts were evaluated by the following two novel probes: a biotinylated and protease (subtilisin Carlsberg)-nicked derivative of theta-toxin and a fluorescein ester of polyethylene glycol-derived cholesterol. Lipid rafts in Abca1-KO macrophages were increased, as demonstrated by both probes. Moreover, activities of nuclear factor kappaB, mRNA and intracellular distribution, and secretion of tumor necrosis factor-alpha (TNF-alpha) were examined after stimulation by lipopolysaccharides (LPSs). LPS-induced responses of the activation of nuclear factor kappaB and TNF-alpha were more prompt and accelerated in the Abca1-KO macrophages compared with wild-type macrophages. Modification of lipid rafts by cyclodextrin and nystatin corrected the abnormal response, suggesting an association between the increased lipid rafts and abnormal TNF-alpha secretion. We report here that Abca1-KO macrophages with defective lipid efflux exhibited increased lipid rafts on the cell surface and accelerated TNF-alpha secretion. PMID- 17079793 TI - Regulation of SR-BI-mediated selective lipid uptake in Chinese hamster ovary derived cells by protein kinase signaling pathways. AB - Scavenger receptor, class B, type I (SR-BI) mediates binding and internalization of a variety of lipoprotein and nonlipoprotein ligands, including HDL. Studies in genetically engineered mice revealed that SR-BI plays an important role in HDL reverse cholesterol transport and protection against atherosclerosis. Understanding how SR-BI's function is regulated may reveal new approaches to therapeutic intervention in atherosclerosis and heart disease. We utilized a model cell system to explore pathways involved in SR-BI-mediated lipid uptake from and signaling in response to distinct lipoprotein ligands: the physiological ligand, HDL, and a model ligand, acetyl LDL (AcLDL). In Chinese hamster ovary derived cells, murine SR-BI (mSR-BI) mediates lipid uptake via distinct pathways that are dependent on the lipoprotein ligand. Furthermore, HDL and AcLDL activate distinct signaling pathways. Finally, mSR-BI-mediated selective lipid uptake versus endocytic uptake are differentially regulated by protein kinase signaling pathways. The protein kinase C (PKC) activator PMA and the phosphatidyl inositol 3-kinase inhibitor wortmannin increase the degree of mSR-BI-mediated selective lipid uptake, whereas a PKC inhibitor has the opposite effect. These data demonstrate that SR-BI's selective lipid uptake activity can be acutely regulated by intracellular signaling cascades, some of which can originate from HDL binding to murine SR-BI itself. PMID- 17079794 TI - Regulation of macrophage apoE secretion and sterol efflux by the LDL receptor. AB - Factors that regulate apolipoprotein E (apoE) secretion by macrophages will have important effects on vessel wall lipid flux and atherosclerosis. Macrophages express the LDL receptor, which binds apoE with high affinity and could thereby affect the net secretion of apoE from macrophages. In these studies, we demonstrate that treatment of J774 macrophages transfected to constitutively express a human apoE3 cDNA with simvastatin, to increase LDL receptor activity, reduces the secretion of apoE. To further examine the relationship between LDL receptor expression and apoE secretion from macrophages, mouse peritoneal macrophages (MPMs) were isolated from mice with constitutively high expression of human LDL receptor to increase overall LDL receptor expression by 2- to 3-fold. Cells with increased LDL receptor expression also showed reduced apoE secretion compared with MPMs with basal LDL receptor expression. The effect of changes in LDL receptor expression on apoE secretion was isoform-specific, with greater reduction of apoE4 compared with apoE3 secretion and no reduction of apoE2 secretion, paralleling the known affinity of each isoform for LDL receptor binding. The effect of the LDL receptor on apoE secretion for each isoform was further reflected in LDL receptor-dependent changes in apoE-mediated cholesterol efflux. These results establish a regulatory interaction between two branches of macrophage sterol homeostatic pathways that could facilitate a rapid response to changes in macrophage sterol content relative to need. PMID- 17079795 TI - Sound-bite research results. PMID- 17079796 TI - The social dynamics of the interview: age, class, and gender. AB - Researchers have paid only limited attention to how social structural factors influence the course and content of interviews. Speech, comportment, and values inherent to gender and other social, structural, and contextual factors, such as age, socioeconomic positioning, and ethnicity, all influence the direction, flow, and content of interviews, informing how we might interpret the information collected in the process. Drawing on interviews conducted within an Australian study on chronic illness and disability, the authors explore the performative nature of the interview and how interviewers and interviewees respond to the structural factors shaping the social dynamics of the interview to produce accounts of social life. PMID- 17079797 TI - "It's no skin off my nose": why people take part in qualitative research. AB - In this article, the authors analyze participants' accounts of why they took part in a repeat-interview study exploring newly diagnosed patients' perceptions of diabetes service provision in Lothian, Scotland. The study involved three semistructured in-depth interviews with each patient (N = 40), which spanned a year. The authors provide a thematic discursive analysis of responses to the question, Can I ask you what made you decide to part in the study and why you've stayed involved over the past year? The main themes are (a) recruitment within health contexts ("the nurse said it would help"), (b) altruism ("if it can help somebody"), (c) qualitative research being seen as inherently innocuous ("nothing to lose"), and (d) therapeutic aspects of interviewing ("getting it off my chest"). The analysis contributes both to the qualitative literature about generic research participation and to a germinal literature exploring qualitative health research participation. PMID- 17079798 TI - From synthesis to script: transforming qualitative research findings for use in practice. AB - Although qualitative research synthesis has secured a place in the evidence-based practice movement, little effort has as yet been directed toward translating the results into material form for practice. The authors transformed a qualitative research synthesis of findings pertaining to stigma in HIV-positive women into a script and DVD. The process of transformation entailed bringing together the norms and imperatives of scientific research, clinical practice, and artistic presentation. PMID- 17079799 TI - Qualitative health research in the era of evidence-based practice. AB - Evidence-based health care (EBHC) sets the tone in health care and health care research nowadays. Qualitative health researchers have to position themselves in a world that is dominated by it. The popularity of EBHC is not due to the rationality of its tenets. In this article, the author addresses major problems in EBHC. Qualitative research is important for providing the understanding that is necessary to apply findings from quantitative research properly and safely. Basic studies about the human experience in illness and regarding human behavior and meaning in general remain of great value, even in the era of EBHC. Qualitative research also plays an important role in developing scholarship. PMID- 17079800 TI - Conducting qualitative interview research with people with expressive language difficulties. AB - Despite the increasing application of qualitative interviewing and analysis approaches, individuals with expressive language difficulties are still frequently excluded from such research. In this article, the authors seek to clarify the role for and importance of conducting qualitative interviews with respondents with impaired expressive language. They review current research with reference to studies conducted with individuals with intellectual disabilities or dementia, or those who have experienced stroke or traumatic brain injury, and identify deficits within existing research. They consider the challenges and difficulties that contribute to the limited inclusion of individuals with impaired expressive communication in qualitative interviews and discuss the way forward with suggestions of possible means of overcoming these obstacles. They argue that a willingness to adapt methods appropriately and to modify expectations is an important factor in ensuring that researchers hear the voice of respondents with expressive language deficits. PMID- 17079801 TI - Photovoice: use of a participatory action research method to explore the chronic pain experience in older adults. AB - Photovoice is a participatory action research method in which individuals photograph their everyday health and work realities. The objective of this study is to determine the efficacy of photovoice as a unique and innovative method examining the pain experiences of clinic and nonclinic-based Black and White adults older than 50 years of age. Participants took photographs and composed written narratives based on their experiences with pain. Respondents also completed several open-ended questions focusing on their daily experiences with chronic pain. Some respondents identified their participation as a way of helping others cope with pain. Others felt it was a way of sharing their experiences with others. Photovoice might be used as an alternative method to enhance the understanding of the pain experience, thereby allowing researchers, health care providers, and policy makers to understand more fully and identify the dimensions of the pain experience not captured through quantitative measures. PMID- 17079802 TI - Draw-and-tell conversations with children about fear. AB - As the demand to include children in research increases, researchers are discovering that few methods exist that are specifically designed with children in mind. In this article, the author introduces the draw-and-tell conversation as a child-centered and child-directed approach to data collection and illustrates its use in a qualitative study of children's fear experiences. Twenty-two children, ages 7 and 8 years, participated. Sequential mixed qualitative analyses suggest that children's draw-and-tell conversations provide new insight into how children describe and experience fear and highlight the unique nature of information accessed when using this approach. PMID- 17079804 TI - The 2006 Henry N. Wagner Lecture: Of mice and men (and positrons)--advances in PET imaging technology. AB - There have been major advances in PET technology that cumulatively have helped improve image quality, increased the range of applications for PET, and contributed to the more widespread use of PET. Examples of these technologic advances include whole-body imaging, 3-dimensional imaging, new scintillator materials, iterative reconstruction algorithms, combined PET/CT, and preclinical PET. New advances on the immediate horizon include the reintroduction of time-of flight PET, which takes advantage of the favorable timing properties of newer scintillators; the integration of PET and MRI scanners into a dual-modality imaging system; and the possibility of further significant improvements in spatial resolution in preclinical PET systems. Sensitivity remains a limiting factor in many PET studies. Although, conceptually, huge gains in sensitivity are still possible, realizing these gains is thwarted largely by economic rather than scientific concerns. Predicting the future is fraught with difficulty; nonetheless, it is apparent that ample opportunities remain for new development and innovation in PET technology that will be driven by the demands of molecular medicine, notably sensitive and specific molecular diagnostic tools and the ability to quantitatively monitor therapeutic entities that include small molecules, peptides, antibodies, nanoparticles, DNA/RNA, and cells. PMID- 17079805 TI - Adjuvant and combined radioimmunotherapy: problems and prospects on the road to Minerva. PMID- 17079806 TI - A comparative regional analysis of coronary atherosclerosis and calcium score on multislice CT versus myocardial perfusion on SPECT. AB - For the noninvasive evaluation of coronary artery disease (CAD), both multislice CT and gated SPECT are available. How these 2 modalities relate, however, is yet unclear. The purpose of this study was to perform a head-to-head comparison of the results of multislice CT and gated SPECT on a regional basis (per vessel distribution territory) in patients with known or suspected CAD. METHODS: One hundred forty patients underwent both multislice CT for coronary calcium scoring and coronary angiography and gated SPECT for myocardial perfusion imaging. The coronary calcium score was determined for each coronary artery. Coronary arteries on multislice CT angiography were classified as having no CAD, insignificant stenosis (<50% luminal narrowing), significant stenosis, or total or subtotal occlusion (>/=90% luminal narrowing). Gated SPECT findings were classified as normal or abnormal (reversible or fixed defects) and were allocated to the territory of one of the various coronary arteries. RESULTS: In coronary arteries with a calcium score of 10 or less, the corresponding myocardial perfusion was normal in 87% (n = 194/224). In coronary arteries with extensive calcifications (score > 400), the percentage of vascular territories with normal myocardial perfusion was lower, 54% (n = 13/24). Similarly, in most of the normal coronary arteries on multislice CT angiography, the corresponding myocardial perfusion was normal on SPECT (156/175, or 89%). In contrast, the percentage of normal SPECT findings was significantly lower in coronary arteries with obstructive lesions (59%) or with total or subtotal occlusions (8%) (P < 0.01). Nonetheless, only 48% of vascular territories with normal perfusion corresponded to normal coronary arteries on multislice CT angiography, whereas insignificant and significant stenoses were present in, respectively, 40% and 12% of corresponding coronary arteries. CONCLUSION: Although a relationship exists between the severity of CAD on multislice CT and myocardial perfusion abnormalities on SPECT, analysis on a regional basis showed only moderate agreement between observed atherosclerosis and abnormal perfusion. Accordingly, multislice CT and gated SPECT provide complementary rather than overlapping information, and further studies should address how these 2 modalities can be integrated to optimize patient management. PMID- 17079807 TI - The prevalence of myocardial ischemia after concurrent chemoradiation therapy as detected by gated myocardial perfusion imaging in patients with esophageal cancer. AB - The detection of myocardial perfusion abnormalities after radiation therapy (RT) has been investigated previously in patients with lymphoma and breast cancer. However, the prevalence and association of such abnormalities with RT in esophageal cancer patients have not been investigated previously. METHODS: The prevalence of myocardial perfusion abnormalities detected using gated myocardial perfusion imaging (GMPI) in patients with esophageal cancer after RT (RT group) was compared with that in patients with esophageal cancer who did not undergo RT (NRT group). The patients' data were extracted from a prospectively collected database. The results of GMPI that were read by multiple readers were tested further by an expert reader who was unaware of the patients' clinical information. This reader's findings were correlated with the different RT isodose lines as seen in the CT for RT planning. Isodose lines containing the affected segments in GMPI as well as the rest of the left ventricle were recorded. Additionally, information with regard to the mean radiation dose to the heart for each patient was collected. An overall, mean radiation dose to the heart in patients with abnormal GMPI studies was compared with that in patients with normal GMPI studies. RESULTS: Fifty-one patients were included, 26 in the RT group and 25 in the NRT group. The mean and median interval between RT and GMPI was 7.5 and 3.0 mo, respectively. We identified myocardial perfusion defects in 14 patients (54%) in the RT group and in 4 patients (16%) in the NRT group. Eleven patients (42%) in the RT group had mild inferior wall ischemia versus only 1 patient (4%) in the NRT group (P = 0.001). All of the patients with inferior wall ischemia had distal esophageal cancer. The remaining 12 patients in the RT group and 21 patients in the NRT group had normal GMPI results. The mean left ventricular ejection fraction was 59.0% +/- 10.7% in the RT group and 59.3% +/- 9.8% in the NRT group (P = not significant). Good agreement was found between the GMPI results interpreted by multiple readers and those of the single expert reader (kappa = 0.84). In 7 of 10 patients (70%) who had abnormal GMPI results in the RT group, the myocardial perfusion defect was encompassed in RT isodose lines >/= 45 Gy, whereas in only 5 of 20 patients (25%) the normal left ventricle was included in the RT isodose line >/= 45 Gy. CONCLUSION: RT is associated with a high prevalence of inferior left ventricular ischemia, as detected using GMPI in patients with distal esophageal cancer. Most perfusion defects are encompassed within an isodose line >/= 45 Gy in the RT plan. PMID- 17079808 TI - Is septal glucose metabolism altered in patients with left bundle branch block and ischemic cardiomyopathy? AB - Left bundle branch block (LBBB) is common in patients with heart failure (HF) and contributes to left ventricular (LV) dysfunction. The abnormal septal motion may alter septal metabolic demand but this has not been well characterized in patients with ischemic cardiomyopathy (ICM) and LV dysfunction. The aim of this study was to determine the effect of LBBB on septal metabolism in patients with ICM, LV dysfunction, and LBBB. METHODS: Fifty-three patients with LV dysfunction and ICM were identified: 34 with LBBB, 19 with normal QRS (30% of this patient population. Absence of this finding was often associated with lateral wall perfusion defects, suggesting an alteration in the metabolic demand on the septum. This may have implications for HF therapies such as resynchronization and requires further study. PMID- 17079809 TI - PET measurement of cardiac and nigrostriatal denervation in Parkinsonian syndromes. AB - Scintigraphic imaging with (123)I-metaiodobenzylguanidine ((123)I-MIBG) has demonstrated extensive losses of cardiac sympathetic neurons in idiopathic Parkinson's disease (IPD). In contrast, normal cardiac innervation has been observed in (123)I-MIBG studies of multiple-system atrophy (MSA) and progressive supranuclear palsy (PSP). Consequently, it has been hypothesized that cardiac denervation can be used to differentiate IPD from MSA and PSP. We sought to test this hypothesis by mapping the distribution of cardiac sympathetic neurons in patients with IPD, MSA, and PSP by using PET and (11)C-meta-hydroxyephedrine ((11)C-HED). Also, the relationship between cardiac denervation and nigrostriatal denervation was investigated by measuring striatal presynaptic monoaminergic nerve density with PET and (11)C-dihydrotetrabenazine ((11)C-DTBZ). METHODS: (11)C-HED and (11)C-DTBZ scans were obtained for patients with IPD (n = 9), MSA (n = 10), and PSP (n = 8) and for age-matched control subjects (n = 10). Global and regional measurements of (11)C-HED retention were obtained to assess the extent of cardiac sympathetic denervation. (11)C-DTBZ binding was measured in the caudate nucleus, anterior putamen, and posterior putamen. RESULTS: As expected, extensive cardiac denervation was observed in several of the patients with IPD. However, substantial cardiac denervation was also seen in some patients with MSA and PSP. (11)C-DTBZ studies demonstrated striatal denervation in all patients with IPD and in most patients with MSA and PSP. No correlation was found between cardiac (11)C-HED retention and striatal (11)C-DTBZ binding. CONCLUSION: Cardiac sympathetic denervation was found to occur not only in IPD but also in other movement disorders, such as MSA and PSP. This finding implies that scintigraphic detection of cardiac sympathetic denervation cannot be used independently to discriminate IPD from other movement disorders, such as MSA and PSP. Cardiac sympathetic denervation was not correlated with striatal denervation, suggesting that the pathophysiologic processes underlying cardiac denervation and striatal denervation occur independently in patients with parkinsonian syndromes. These findings provide novel information about central and peripheral denervation in patients with neurodegenerative disorders. PMID- 17079810 TI - Hypometabolism exceeds atrophy in presymptomatic early-onset familial Alzheimer's disease. AB - The aim of the present study is to compare brain atrophy with hypometabolism as preclinical markers of Alzheimer's disease (AD) by studying presymptomatic individuals from families with known early-onset autosomal dominant AD (FAD) carrying mutations in the Presenilin 1 gene. METHODS: Seven asymptomatic at-risk FAD individuals (age, 35-49 y; 4 women; education >/= 12 y) and 7 matched healthy control subjects received complete clinical, neuropsychologic, MRI, and (18)F-FDG PET examinations. Regions of interest (ROIs: whole brain [WB], hippocampus [Hip], entorhinal cortex [EC], posterior cingulate cortex [PCC], inferior parietal lobule [IPL], and superior temporal gyrus (STG]) were drawn on the MRI scans of all subjects and used to measure volumes on MRI and glucose metabolism (MRglc) from the MRI-coregistered, atrophy-corrected PET scans. RESULTS: Compared with controls and after correcting for head size, MRI volume reductions in FAD subjects were restricted to the IPL (18%, P < 0.02). After atrophy correction and adjusting for pons MRglc, PET MRglc reductions were found in all FAD subjects compared with controls in the WB (13%), bilaterally in the IPL (17%) and in the STG (12%), and in the left EC (21%), PCC (20%), and Hip (12%) (P values < 0.05). PET MRglc measurements were consistently less variable than MRI measures, yielding significantly larger effect sizes in separating FAD from controls. CONCLUSION: Presymptomatic FAD individuals show widespread MRglc reductions consistent with the typical AD PET pattern in the relative absence of structural brain atrophy. These data further suggest that PET MRglc measures may serve as biomarkers for the preclinical diagnosis of AD. PMID- 17079811 TI - Quantification of protein synthesis in the human brain using L-[1-11C]-leucine PET: incorporation of factors for large neutral amino acids in plasma and for amino acids recycled from tissue. AB - The rate of incorporation of exogenous amino acids into brain proteins is indicative of the protein synthesis rate (PSR). The objective of this study was to assess the effect of plasma concentrations of leucine and large neutral amino acids (LNAAs) on the unidirectional uptake rate constant (Kcplx) of l-[1-(11)C] leucine in the brain and to estimate the amino acid pool recycled from tissue. METHODS: Twenty-seven healthy adult volunteers (11 men and 16 women; age range, 20-50 y) underwent dynamic l-[1-(11)C]-leucine PET with arterial blood sampling. Data were analyzed with a standard 2-tissue-compartment model yielding the unidirectional uptake rate of plasma leucine into tissue (Kcplx = K(1)k(3)/(k(2) + k(3))) and the fraction of leucine originating from exogenous sources (lambda = k(2)/(k(2) + k(3))). PSR in brain was calculated as PSR = [Kcplx/lambda] x leucine. RESULTS: The mean plasma concentration of the sum of all LNAAs was 13% higher in men (981 +/- 86 micromol/L) than in women (850 +/- 76 micromol/L, P = 0.012), whereas the plasma leucine concentration was found to be similar in both sexes (men, 64 +/- 20 micromol/L; women, 58 +/- 21 micromol/L, P = 0.57). The whole-brain value for lambda was determined to be 0.64 +/- 0.03 and did not show a sex difference (P = 0.66). Whole-brain Kcplx values were significantly higher in women (0.0162 +/- 0.0024) than in men (0.0121 +/- 0.0031, P = 0.011); however, after normalization of the Kcplx to a standard plasma concentration of the sum of all LNAAs (Kcplx'), the Kcplx' was similar between the sexes (P = 0.21), as was the PSR' (1.24 +/- 0.49 micromol/L/min in men; 1.29 +/- 0.62 micromol/L/min in women, P = 0.87). No relationship between plasma leucine and Kcplx (r = -0.13, P = 0.63) was observed. Finally, there was a significant correlation between the PSR and the Kcplx derived using Patlak graphical analysis (rho = 0.65, P < 0.001). CONCLUSION: We conclude that both the Kcplx macroparameter and the PSR are stable indices of brain protein synthesis and are appropriate measures for testing altered protein synthesis in neurologic disorders. PMID- 17079812 TI - Metabolite considerations in the in vivo quantification of serotonin transporters using 11C-DASB and PET in humans. AB - PET studies of the serotonin (5-hydroxytryptamine, or 5-HT) transporter are increasingly using (11)C-3-amino-4-(2 dimethylaminomethylphenylsulfanyl)benzonitrile (DASB). We noted that the percentage of unmetabolized (11)C-DASB is often lower at 2 min after injection than at 12 min. We hypothesized that this is due to initial "trapping" of the unmetabolized (11)C-DASB compound in the lung, a major 5-HT transporter site and dose-limiting organ. To determine whether binding to an extracranial pool of 5-HT transporters contributes to the lower initial level of unmetabolized (11)C-DASB, we examined the effects of sertraline. METHODS: Eleven healthy volunteers had 2 (11)C-DASB PET scans on the same day, and 6 of the 11 had a third scan after sertraline administration. The unmetabolized (11)C-DASB fraction was measured in arterial plasma as a function of time and was fit with 2 exponentials with no damping, power function damping, or no damping with the first point removed. RESULTS: Power function damping best fit the data as assessed by visual inspection and residuals and resulted in greater distribution volumes than did no damping with the first point removed. Test-retest reproducibility improved when power function damping was used, as compared with no damping with the first point removed. Oral sertraline raised the 2-min unmetabolized (11)C-DASB percentage. CONCLUSION: Measurement and fitting of early metabolism time points improves curve fitting, significantly affects volume-of-distribution determination, and improves test-retest reproducibility. Saturation of lung 5-HT transporters by sertraline prevents the initial trapping of (11)C-DASB. Initial trapping of high affinity radioligands may be important in the quantification of the binding of other ligands with a high concentration of binding sites in the lungs. PMID- 17079813 TI - Forced diuresis improves the diagnostic accuracy of 18F-FDG PET in abdominopelvic malignancies. AB - Our aim was to evaluate the role of forced diuresis in improving the diagnostic accuracy of abdominopelvic (18)F-FDG PET. METHODS: Thirty-two patients were enrolled. Besides the presence of known intravesical tumors or undefined renal lesions on the initial PET scan, the inclusion criterion was the appearance of indeterminate or equivocal (18)F-FDG foci that extended along the course of the urinary tract and could not confidently be separated from urinary activity. For each patient, a second abdominopelvic PET study was performed after intravenous injection of 0.5 mg of furosemide per kilogram of body weight (maximum, 40 mg) coupled with parenteral infusion of physiologic saline. RESULTS: Forced diuresis coupled with parenteral hydration eliminated any significant (18)F-FDG activity from the lower urinary tract in 31 (97%) of 32 patients after the bladder had been voided 3 successive times. Twelve intravesical lesions were visualized with outstanding clarity, whereas radiologic suspicion of locally recurrent bladder tumors was ruled out in 3 patients. Among 14 indeterminate or equivocal extravesical foci, 7 were deemed of no clinical value because they disappeared after furosemide challenge, whereas 7 persisting foci were proven to be true positive PET findings. The performance of (18)F-FDG PET in characterizing 3 renal space-occupying lesions could not be improved by our protocol. CONCLUSION: Furosemide challenge has the potential to noninvasively resolve the inherent (18)F-FDG contrast handicap in the lower urinary tract. PMID- 17079814 TI - 18F-FDG PET definition of gross tumor volume for radiotherapy of non-small cell lung cancer: is a single standardized uptake value threshold approach appropriate? AB - PET with (18)F-FDG has been used in radiation treatment planning for non-small cell lung cancer (NSCLC). Thresholds of 15%-50% the maximum standardized uptake value (SUV(max)) have been used for gross tumor volume (GTV) delineation by PET (PET(GTV)), with 40% being the most commonly used value. Recent studies indicated that 15%-20% may be more appropriate. The purposes of this study were to determine which threshold generates the best volumetric match to GTV delineation by CT (CT(GTV)) for peripheral NSCLC and to determine whether that threshold can be generalized to tumors of various sizes. METHODS: Data for patients who had peripheral NSCLC with well-defined borders on CT and SUV(max) of greater than 2.5 were reviewed. PET/CT datasets were reviewed, and a volume of interest was determined to represent the GTV. The CT(GTV) was delineated by using standard lung windows and reviewed by a radiation oncologist. The PET(GTV) was delineated automatically by use of various percentages of the SUV(max). The PET(GTV)-to CT(GTV) ratios were compared at various thresholds, and a ratio of 1 was considered the best match, or the optimal threshold. RESULTS: Twenty peripheral NSCLCs with volumes easily defined on CT were evaluated. The SUV(max) (mean +/- SD) was 12 +/- 8, and the mean CT(GTV) was 198 cm(3) (97.5% confidence interval, 5-1,008). The SUV(max) were 16 +/- 5, 13 +/- 9, and 3.0 +/- 0.4 for tumors measuring greater than 5 cm, 3-5 cm, and less than 3 cm, respectively. The optimal thresholds (mean +/- SD) for the best match were 15% +/- 6% for tumors measuring greater than 5 cm, 24% +/- 9% for tumors measuring 3-5 cm, 42% +/- 2% for tumors measuring less than 3 cm, and 24% +/- 13% for all tumors. The PET(GTV) at the 40% and 20% thresholds underestimated the CT(GTV) for 16 of 20 and 14 of 20 lesions, respectively. The mean difference in the volumes (PET(GTV) minus CT(GTV) [PET(GTV) - CT(GTV)]) at the 20% threshold was 79 cm(3) (97.5% confidence interval, -922 to 178). The PET(GTV) at the 20% threshold overestimated the CT(GTV) for all 4 tumors measuring less than 3 cm and underestimated the CT(GTV) for all 6 tumors measuring greater than 5 cm. The CT(GTV) was inversely correlated with the PET(GTV) - CT(GTV) at the 20% threshold (R(2) = 0.90, P < 0.0001). The optimal threshold was inversely correlated with the CT(GTV) (R(2) = 0.79, P < 0.0001). CONCLUSION: No single threshold delineating the PET(GTV) provides accurate volume definition, compared with that provided by the CT(GTV), for the majority of NSCLCs. The strong correlation of the optimal threshold with the CT(GTV) warrants further investigation. PMID- 17079815 TI - Sequential 99mTc-hydrazinonicotinamide-annexin V imaging for predicting response to chemotherapy. AB - This study was undertaken to evaluate changes in relative (99m)Tc hydrazinonicotinamide (HYNIC)-annexin V tumor uptake over time in patients undergoing chemotherapeutic treatment at baseline and at 5-7 h and 40-44 h after treatment initiation. Imaging results are related to clinical outcomes, as assessed with response evaluation criteria in solid tumors (RECIST). METHODS: We prospectively included 20 patients (11 men and 9 women; mean age, 59.8 y; range, 22-75 y) scheduled for chemotherapy (n = 19) or bisphosphonate treatment (n = 1). Curable disease was present in 5 patients. The other patients had metastatic disease and were treated in a palliative setting. Three of the 20 enrolled patients were excluded from analysis: 1 patient ultimately refused the proposed chemotherapy treatment; because of difficulties with the labeling procedure, 1 patient did not receive a pretreatment scan; and 1 patient presented with an allergic reaction (rash and nausea) to the (99m)Tc-HYNIC-annexin V formulation. The remaining 17 patients underwent 3 scintigraphic scans with (99m)Tc-HYNIC annexin V: before treatment and 5-7 h and 40-44 h after treatment initiation. The tumor response was evaluated with RECIST and related to observed changes in the ratios of tumor activity to background activity for the largest known lesion; values exceeding 25% the baseline value on either the 5- to 7-h scan or the 40- to 44-h scan were considered significant. RESULTS: With the proposed sequential imaging protocol and a 25% change threshold, responders to treatment could be separated from nonresponders with a 94% accuracy (16/17 patients). CONCLUSION: Sequential (99m)Tc-HYNIC-annexin V imaging may allow for assessment of the response to chemotherapy within 3 d after treatment initiation. PMID- 17079816 TI - Radionuclide investigations of the urinary tract in the era of multimodality imaging. AB - This article presents the role of nuclear medicine procedures in investigating renal and parenchymal disease, as well as upper urinary tract abnormalities. More specifically, the use of scintigraphy is described in the exploration of urinary tract dilatation and UTIs, vesicoureteric reflux, renovascular hypertension, and renal transplants. With a low radiation burden and the absence of sedation, these nuclear medicine procedures are easy to perform and can provide clinicians with valuable data on renal perfusion and the function of individual kidneys, as well as on urinary tract dynamics. However, knowledge of limitations and technical pitfalls is essential in understanding the role of scintigraphy among contemporary imaging methods and the unique information it supplies in nephrourology. PMID- 17079817 TI - Accurate noninvasive measurement of infarct size in mice with high-resolution PET. AB - Reliable, repeatable, and time-efficient noninvasive measurement of infarct size in mice with PET would benefit studies aimed at the exploration of biochemical and functional changes associated with acute myocardial infarction (MI). PET with the radioactively labeled glucose derivative (18)F-FDG is used in humans to distinguish between viable but dysfunctional and nonviable myocardium. In this study, the feasibility, accuracy, and time efficiency of (18)F-FDG PET for quantification of infarct size in mice using a high-resolution animal PET device was evaluated in comparison with histomorphometry. METHODS: Mice were subjected to surgery with permanent ligation of the left anterior descending artery. PET was performed before and 7 d after surgery. The infarct size was determined from the PET studies using both manual and automated delineation. The second PET scan was followed by histomorphometric analysis. RESULTS: An excellent correlation between PET and histomorphometry was found for both manual (R = 0.98) and automated (R = 0.98) delineation, with linear regression curves close to unity (manual: y = 1.10x - 0.01; automated: y = 1.12x - 0.02). Automated analysis required <1 min per study. CONCLUSION: The measurement of infarct size in mice with (18)F-FDG PET is feasible and highly accurate. This noninvasive methodology permits unique longitudinal studies of biochemical parameters in mice and facilitates studies that aim to assess the effect of surgical and pharmacologic intervention after acute MI. PMID- 17079818 TI - Application of 18F-FDG PET for monitoring the therapeutic effect of antiinflammatory drugs on stabilization of vulnerable atherosclerotic plaques. AB - The rupture of atherosclerotic vulnerable plaques and subsequent formation of thrombi are the main factors responsible for myocardial and cerebral infarctions. Because macrophage infiltration plays an essential role in plaque rupturing, pharmacologic therapy that reduces macrophage infiltration is required to stabilize the vulnerable plaques. The monitoring of therapeutic effect is important in assessing the therapeutic effects of drugs for individual patients. We previously reported that (18)F-FDG accumulates in macrophage-rich plaques. The present study was undertaken to investigate the usefulness of (18)F-FDG PET for monitoring therapies that target vascular inflammation. METHODS: Myocardial infarction-prone Watanabe heritable hyperlipidemic rabbits were used in this study. The antioxidant probucol was included in the diet fed to 4 rabbits starting at 10 mo of age (probucol group). In a control study, 4 rabbits received standard rabbit chow (control group). (18)F-FDG PET experiments were performed on both groups before the study and at 1, 3, and 6 mo after treatment. After the last imaging session, the rabbits were sacrificed at 3 h after injection of (18)F FDG, and the aortas were removed. The accumulated radioactivity was then measured, and the number of macrophages was determined by examination of stained sections. RESULTS: At the age of 10 mo, before the treatment, the aorta could be imaged by (18)F-FDG PET in all rabbits. The aorta could not be imaged after 6 mo of probucol treatment, whereas intense radioactivity was observed in the control rabbits throughout the investigation. The standardized uptake values (SUVs) of the aorta were decreased significantly in the probucol group after 3 mo of intervention as compared with the pretreatment period. The SUVs of the control group were increased gradually at 6 mo. Radioactivity in the aorta was significantly lower in the probucol group than that in the control group. Macrophages were already present at the beginning of the study, and probucol treatment for 6 mo resulted in a significant reduction of macrophage infiltration. CONCLUSION: (18)F-FDG PET was able to image the reduction of inflammation by probucol. (18)F-FDG PET should be useful for evaluating the therapeutic effect of drugs clinically and for the development of new drugs that can stabilize vulnerable plaques. (18)F-FDG PET should be useful for evaluating the therapeutic effect of drugs clinically and for the development of new drugs that can reduce inflammation of vulnerable plaques. PMID- 17079819 TI - Evidence that myocardial Na/I symporter gene imaging does not perturb cardiac function. AB - Adenoviral Na/I symporter (NIS) gene transfer has emerged as a promising method for myocardial gene imaging but concern over possible perturbation of cardiac function persists. In this study, we addressed this issue with cultured cardiac cells and serial echocardiography, creatine kinase (CK) measurements, and histologic examination of rats intramyocardially injected with an adenovirus that expresses both NIS and enhanced green fluorescent protein (EGFP) (Ad.EGFP.NIS) or a control virus (Ad.EGFP). METHODS: H9C2 cardiac myoblasts differentiated into cardiomyocytes were evaluated for the effect of Ad.EGFP.NIS and Ad.EGFP infection on viable cell number and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Rats injected intramyocardially with 3 x 10(8) plaque forming units of Ad.EGFP.NIS (n = 9) or Ad.EGFP (n = 8) underwent serial echocardiographic measurements of heart rate, left ventricular (LV) dimensions, ejection fraction (EF), and fractional shortening (FS) on the day before and on days 4 and 9 after gene transfer. Five Ad.EGFP.NIS rats also underwent repeated (123)I imaging from which ratios of cardiac to mediastinal counts (C/M ratios) were obtained. Separate rats underwent serial measurements of serum CK, myocardial myeloperoxidase assays, and microscopic assessment of inflammation. RESULTS: Cultured cardiac cells showed no change in cell viability or proliferation at 4 and 9 d after Ad.EGFP.NIS or Ad.EGFP infection compared with controls. (123)I scintigraphy demonstrated high cardiac radiouptake at Ad.EGFP.NIS injection sites by days 2 and 4 (C/M ratios, 5.0 +/- 0.6 and 5.1 +/- 1.0, respectively), followed by a complete loss of uptake by day 9 (C/M ratio, 1.4 +/- 0.0). Serial echocardiography revealed no difference in heart rate, LV dimensions, or functional parameters between Ad.EGFP.NIS and Ad.EGFP groups at any given time. Mild reductions in LVEF and LVFS by day 9 compared with baseline were similar for both Ad.EGFP (88.2% +/- 6.4% vs. 79.6% +/- 5.0% for LVEF and 0.55 +/- 0.10 vs. 0.44 +/- 0.05 for LVFS) and Ad.EGFP.NIS groups (88.0% +/- 5.4% vs. 78.7% +/- 4.6% for LVEF and 0.54 +/- 0.09 vs. 0.42 +/- 0.05 for LVFS). Serial serum CK and myocardial myeloperoxidase activities were not elevated in either group, in contrast to substantial increases found after ischemia-reperfusion injury. Histology revealed similar mild inflammatory cell infiltration restricted to the injection site for both groups. CONCLUSION: The results of this study demonstrate that myocardial NIS gene imaging does not cause significant myocardial injury or perturbed cardiac function, other than mild effects likely due to adenoviral vector-associated host response. Thus, this practical and convenient reporter gene strategy can be used safely for noninvasive myocardial gene imaging in living subjects. PMID- 17079820 TI - Construction and evaluation of multitracer small-animal PET probabilistic atlases for voxel-based functional mapping of the rat brain. AB - Automated voxel-based or predefined volume-of-interest (VOI) analysis of rodent small-animal PET data is necessary for optimal use of information because the number of available resolution elements is limited. We have mapped metabolic ((18)F-FDG), dopamine transporter (DAT) (2'-(18)F-fluoroethyl(1R-2-exo-3-exe)-8 methyl-3-(4-chlorophenyl)-8-azabicyclo[3.2.1]-octane-2-carboxylate [(18)F-FECT]), and dopaminergic D(2) receptor ((11)C-raclopride) small-animal PET data onto a 3 dimensional T2-weighted MRI rat brain template oriented according to the rat brain Paxinos atlas. In this way, ligand-specific templates for sensitive analysis and accurate anatomic localization were created. Registration accuracy and test-retest and intersubject variability were investigated. Also, the feasibility of individual rat brain statistical parametric mapping (SPM) was explored for (18)F-FDG and DAT imaging of a 6-hydroxydopamine (6OHDA) model of Parkinson's disease. METHODS: Ten adult Wistar rats were scanned repetitively with multitracer small-animal PET. Registrations and affine spatial normalizations were performed using SPM2. On the MRI template, a VOI map representing the major brain structures was defined according to the stereotactic atlas of Paxinos. (18)F-FDG data were count normalized to the whole-brain uptake, whereas parametric DAT and D(2) binding index images were constructed by reference to the cerebellum. Registration accuracy was determined using random simulated misalignments and vectorial mismatching. RESULTS: Registration accuracy was between 0.24 and 0.86 mm. For (18)F-FDG uptake, intersubject variation ranged from 1.7% to 6.4%. For (11)C-raclopride and (18)F-FECT data, these values were 11.0% and 5.3%, respectively, for the caudate-putamen. Regional test-retest variability of metabolic normalized data ranged from 0.6% to 6.1%, whereas the test-retest variability of the caudate-putamen was 14.0% for (11)C-raclopride and 7.7% for (18)F-FECT. SPM analysis of 3 individual 6OHDA rats showed severe hypometabolism in the ipsilateral sensorimotor cortex (P C and -1282G>A) showed a marginal association with total IgE levels (0.03 and 0.03 in recessive models, respectively). Our findings suggest that polymorphisms in RUNX1 might be one of the genetic factors for the regulation of IgE production. PMID- 17079876 TI - The medical genetics workforce: an analysis of clinical geneticist subgroups. AB - PURPOSE: Clinical geneticists with a Doctor of Medicine degree face challenges to meet the growing population demand for genetic services. This study was designed to assist the profession with workforce planning by identifying clinically relevant subgroups of geneticists and describing their professional characteristics and clinical practices. Geneticists' patient care productivity is compared across subgroups and other medical specialists. METHODS: Part of a comprehensive national study of genetic services and the health workforce, this study uses data from a 2003 survey of geneticists certified by the American Board of Medical Genetics. This study includes 610 clinical geneticists who spend at least 5% of their time in direct patient-care services. An iterative approach was used to identify five subgroups based on the types of new patients seen. We conducted a descriptive analysis of subgroups by demographic, training, professional, and practice characteristics. RESULTS: The subgroups include general (36%), pediatric (28%), reproductive (15%), metabolic (14%), and adult (7%) geneticists. Clinically relevant variations across subgroups were noted in training, professional, and practice parameters. Subgroups vary across patient care hours (median, 15-33 hours/week) and total weekly work hours (52-60 hours). New patient visits (mean, 222-900/year) are higher than follow-up patient visits (mean, 155-405) for all subgroups except metabolic geneticists. CONCLUSION: Although many geneticists practice as generalist geneticists, this study provides an evidence base for distinguishing clinically relevant subgroups of geneticists. Geneticists provide similar numbers of new patient visits and far fewer follow-up visits than other medical specialists. These findings are relevant to geneticist workforce planning. PMID- 17079877 TI - Quantity versus quality: optimal methods for cell-free DNA isolation from plasma of pregnant women. AB - PURPOSE: Methods to isolate cell-free fetal DNA from maternal plasma are critical in developing noninvasive fetal DNA testing strategies. Given that plasma consists of heterogeneous DNA-size fragments in a complex mix of proteins, recovery and analysis of this DNA are understandably inefficient. To facilitate recovery, we performed qualitative and quantitative analysis of DNA isolated from maternal plasma. METHODS: DNA isolated from maternal blood (n = 15) was compared using five different DNA isolation protocols: two conventional, two column-based, and one magnetic-bead based. Purity and concentration of DNA recovered were determined with a NanoDrop spectrophotometer. Real-time polymerase chain reaction quantification of the beta-globin and DYS1 loci was performed to determine total and fetal-specific genome equivalents, respectively. RESULTS: DNA quality and quantity were different among the five methods tested. Although purity and concentration of total DNA were greatest with the conventional boiling-lysis approach, correct detection of a male fetus was achieved in only 62.5% of cases. DNA isolation using the magnetic beads yielded the highest quantity of total DNA (2018.83 +/- 4.09 GEq/mL), with 100% fetal DNA detection. CONCLUSIONS: Optimal plasma DNA recovery protocols must take into account DNA purity and concentration. We confirm that the magnetic-beads method provides a fast, simple, sensitive, and specific approach to purify plasma DNA. The resulting high-quality DNA facilitates efficient examination of fetal DNA sequences. PMID- 17079878 TI - Chip-based mtDNA mutation screening enables fast and reliable genetic diagnosis of OXPHOS patients. AB - PURPOSE: Oxidative phosphorylation is under dual genetic control of the nuclear and the mitochondrial DNA (mtDNA). Oxidative phosphorylation disorders are clinically and genetically heterogeneous, which makes it difficult to determine the genetic defect, and symptom-based protocols which link clinical symptoms directly to a specific gene or mtDNA mutation are falling short. Moreover, approximately 25% of the pediatric patients with oxidative phosphorylation disorders is estimated to have mutations in the mtDNA and a standard screening approach for common mutations and deletions will only explain part of these cases. Therefore, we tested a new CHIP-based screening method for the mtDNA. METHODS: MitoChip (Affymetrix) resequencing was performed on three test samples and on 28 patient samples. RESULTS: Call rates were 94% on average and heteroplasmy detection levels varied from 5-50%. A genetic diagnosis can be made in almost one-quarter of the patients at a potential output of 8 complete mtDNA sequences every 4 days. Moreover, a number of potentially pathogenic unclassified variants (UV) were detected. CONCLUSIONS: The availability of long-range PCR protocols and the predominance of single nucleotide substitutions in the mtDNA make the resequencing CHIP a very fast and reliable method to screen the complete mtDNA for mutations. PMID- 17079879 TI - Molecular diagnosis of Beckwith-Wiedemann syndrome using quantitative methylation sensitive polymerase chain reaction. AB - PURPOSE: Beckwith-Wiedemann Syndrome is caused by defects in imprinted gene expression at 11p15. Currently, quantitative Southern analysis using DNA methylation-sensitive restriction enzymes is used in molecular diagnosis of this syndrome. METHODS: We describe a rapid and highly quantitative test for assessing DNA methylation at 11p15 using sodium bisulfite treatment of genomic DNA coupled with quantitative TaqMan methylation-sensitive polymerase chain reaction. RESULTS: TaqMan MSP can assess DNA methylation at both differentially methylated region (DMR)1 and DMR2 at 11p15. In addition, by using TaqMan MSP we were able to determine the parent of origin of a duplication of 11p15 by quantification of both DMR1 and DMR2 DNA methylation. CONCLUSION: TaqMan MSP method is a robust and rapid method for detecting changes in DNA methylation that compares favorably to the current standard of Southern blot for DNA methylation analysis. Assessment of DMR1 and DMR2 provides the most comprehensive assay for methylation defects in Beckwith Wiedemann Syndrome, accounting for more than 70% of the cases. The advantages of TaqMan MSP are that it requires less DNA and that it is rapid, less labor-intensive, and amenable to high-throughput analysis. Moreover, this approach can be modified to assess DNA methylation changes anywhere in the genome. PMID- 17079880 TI - Characterization of an unusual deletion of the galactose-1-phosphate uridyl transferase (GALT) gene. AB - PURPOSE: We previously reported a deletion of the Galactose-1-Phosphate Uridyl Transferase (GALT) gene. This deletion can cause apparent homozygosity for variants located on the opposite allele, potentially resulting in a discrepancy between the biochemical phenotype and the apparent genotype in an individual. The purpose of this study was to determine the deletion breakpoints, allowing the development of a rapid and reliable molecular test for the mutation. METHODS: A Polymerase Chain Reaction walking strategy was used to map the 5' and 3' breakpoints. The junction fragment was amplified and sequenced to precisely characterize the deletion breakpoints. RESULTS: The deletion has a bipartite structure involving two large segments of the GALT gene, while retaining a short internal segment of the gene. Molecular characterization allowed the development of a deletion specific Polymerase Chain Reaction-based assay. In 25 individuals who had a biochemical carrier galactosemia phenotype, but tested negative for 8 common GALT gene variants, 3 carried this deletion. CONCLUSION: This deletion occurs at an appreciable frequency and should be considered when there is a discrepancy between the genotype and biochemical phenotype. Many of the individuals carrying the allele were of Ashkenazi Jewish ancestry suggesting that the deletion may be a common cause of galactosemia in that population. PMID- 17079881 TI - Molecular bases of hearing loss in multi-systemic mitochondrial cytopathy. AB - PURPOSE: Hearing loss is a common clinical feature in classic mitochondrial syndromes. The purpose of this study was to evaluate the diverse molecular etiologies and natural history of hearing loss in multi-systemic mitochondrial cytopathies and the possible correlation between degree of hearing loss and neurological phenotype. METHODS: In this retrospective study we evaluated the clinical features and molecular bases of hearing loss associated with multi systemic mitochondrial cytopathy. Forty-five patients with sensorineural hearing loss and definite diagnosis of mitochondrial cytopathy according to the published diagnostic criteria were studied. RESULTS: The sensorineural hearing loss was progressive and for the most part symmetrical with involvement of the higher frequencies. Both cochlear and retrocochlear involvement were found in this cohort. No correlation was found between the degree of hearing loss and the number and severity of neurological manifestations. Deleterious mtDNA point mutations of undisputed pathogenicity were identified in 18 patients. The A3243G mutation was the most frequently encountered among this group. MtDNA depletion, over-replication, and multiple deletions were found in further 11 cases. CONCLUSION: This study reveals an expanding spectrum of mtDNA abnormalities associated with hearing loss. No correlation was found between the degrees of hearing loss and the severity of neurological manifestations. PMID- 17079882 TI - The role of hereditary nonpolyposis colorectal cancer in the management of familial ovarian cancer. AB - PURPOSE: Familial ovarian cancer is most often associated with hereditary breast and ovarian cancer, implicating mutations in the BRCA1 and BRCA2 genes. Hereditary nonpolyposis colorectal cancer, another common syndrome, is also associated with ovarian cancer and is caused by DNA mismatch repair genes. We sought to identify the role of hereditary nonpolyposis colorectal cancer in women with family histories of ovarian cancer. METHODS: The likelihood of a genetic syndrome in 226 oophorectomized women in the Gilda Radner Familial Ovarian Cancer Registry was determined by pedigree analysis using clinical criteria and by calculating the probability of a mutation in genes responsible for hereditary breast and ovarian cancer and hereditary nonpolyposis colorectal cancer using available risk models. RESULTS: Some 86% had a BRCA gene mutation likelihood of 7.8% or higher, warranting consideration of hereditary breast and ovarian cancer. Of the 32 women below this threshold, 4 (12.5%) had family histories that met criteria for clinical diagnosis of hereditary nonpolyposis colorectal cancer. In addition, 16 women (7%) with a BRCA mutation likelihood greater than 7.8% met clinical criteria for hereditary nonpolyposis colorectal cancer or warranted its inclusion in the differential diagnosis. Among all study respondents, 9% had family histories warranting consideration of hereditary nonpolyposis colorectal cancer. CONCLUSION: Hereditary nonpolyposis colorectal cancer should be considered in the differential diagnosis of women with family histories of ovarian cancer. PMID- 17079883 TI - Self-rated breast cancer risk among women reporting a first-degree family history of breast cancer on office screening questionnaires in routine medical care: the role of physician-delivered risk feedback. AB - PURPOSE: We investigated whether risk-related feedback delivered by one's primary care physician is associated with self-ratings of risk among women found to have a first-degree family history of breast cancer on office screening questionnaires. METHODS: DESIGN: Mailed survey of women registered with the Cancer Genetics Network having a first-degree family history of breast cancer. Eligibility: Completion of primary care-based family history screening within the past year. INDEPENDENT VARIABLE: presence of physician feedback about breast cancer risk. Dependent variable: self-rated breast cancer risk. Modifying variable: trust in one's doctor. RESULTS: Three hundred one women met eligibility criteria (73% minimum response rate); feedback was associated with rating one's risk to be "high" in both crude and multivariate analysis. (ORadj = 2.38; 95% CI = 1.30, 4.38). Higher levels of trust in the physician were associated in a dose dependent fashion with the strength of association between feedback and self rating one's risk to be high. CONCLUSIONS: Physician feedback following the identification of a first-degree family history of breast cancer appears to influence whether or not women categorize themselves to be at high risk and trust is an important modifier of this association. PMID- 17079884 TI - Maternal understanding of infant diabetes risk: differential effects of maternal anxiety and depression. AB - PURPOSE: This study describes maternal understanding of infant risk associated with newborn genetic screening for type 1 diabetes. METHODS: Mothers of at-risk infants (n = 195), identified through the Prospective Assessment of Newborns for Diabetes Autoimmunity study, were notified of risk status by standardized script. Mothers participated in structured telephone interviews 1 and 3.5 months after notification that assessed understanding of infant risk and psychologic response to the news. RESULTS: Most mothers (78.5%) were accurate in their understanding of infant risk at the initial interview, with a slight decline at the follow-up interview (73%). There was a significant increase in underestimation of risk from the initial (12%) to the follow-up interview (19%) (chi2 (1) = 6.0, P = .01). Mothers with less education, those from ethnic minority backgrounds, and those who were not married tended to be less accurate. Further, mothers who experienced more anxiety and fewer depressive symptoms in response to the news were more likely to be accurate. Likewise, underestimation of risk was associated with fewer anxiety and more depressive symptoms. CONCLUSION: This study highlights the complex picture of factors promoting maternal understanding of infant diabetes risk in a sample of mothers whose newborns had been identified as at increased risk for type 1 diabetes. PMID- 17079885 TI - Fetal gender and maternal serum screening markers. PMID- 17079895 TI - Focusing on JCAHO National Patient Safety Goals. PMID- 17079896 TI - Giving thrush a thrashing. PMID- 17079897 TI - Cables and electrodes can burn patients during MRI. PMID- 17079898 TI - Managing osteoarthritis pain. PMID- 17079899 TI - What's a fat embolism? PMID- 17079917 TI - Understanding the 12-lead ECG, part I. PMID- 17079919 TI - Keeping warfarin therapy in balance. PMID- 17079920 TI - Patient education series. Coumadin (warfarin) therapy. PMID- 17079921 TI - Assessing the cranial nerves. PMID- 17079922 TI - View from the cockpit: what the airline industry can teach us about patient safety. PMID- 17079923 TI - Before you give that vaccination... PMID- 17079924 TI - Managing symptoms for a "good death". PMID- 17079929 TI - Myths and facts...about implantable cardioverter defibrillators. PMID- 17079931 TI - Tension pneumothorax from central line placement. PMID- 17079932 TI - Pancreas enters its third decade. PMID- 17079933 TI - Confusion in the imaging ranks: time for a change? PMID- 17079934 TI - Trends in the epidemiology of the first attack of acute pancreatitis: a systematic review. AB - OBJECTIVE: To systematically review trends in the epidemiology of the first attack of acute pancreatitis (AP) based on reported population-based studies. METHODS: From Medline, we retrieved 18 full-length English language peer-reviewed original articles published from 1966 to June 2005 with population-based information on the epidemiology of first-attack AP. Trends over time were analyzed based on study- and country-specific data and the study site (UK vs non UK). RESULTS: Eight studies were from the UK and 10 from other European centers. An increase in the annual incidence per 100,000 for first-attack AP was reported in 10 of 12 studies with longitudinal data from 4 countries (UK, Sweden, Denmark, and Netherlands). The overall AP incidence seems to be higher in non-UK studies compared with that of UK, partly explained by a higher incidence of alcoholic pancreatitis in non-UK studies. A linear trend for increase in gallstone pancreatitis incidence over time was observed irrespective of the study site (UK or non-UK). The AP incidence and mortality increased with age. Gallstone pancreatitis was more common in female subjects, and alcoholic pancreatitis was more common in middle-aged male subjects. The AP case fatality (%) has decreased over time, but the overall population mortality rate per 100,000 has remained unchanged. Recurrence after the first attack is milder with a substantially lower mortality. CONCLUSIONS: The incidence of AP seems to be increasing. Differences in the incidence and etiology between and within countries reflect differences in the risk factor prevalence. Case-fatality rate, but not the population-based mortality rate, decreased over time. PMID- 17079935 TI - Describing computed tomography findings in acute necrotizing pancreatitis with the Atlanta classification: an interobserver agreement study. AB - OBJECTIVES: The 1992 Atlanta classification is a clinically based classification system that defines the severity and complications of acute pancreatitis. A study was undertaken to assess the interobserver agreement of categorizing peripancreatic collections on computed tomography (CT) using the Atlanta classification. METHODS: Preoperative contrast-enhanced CTs from 70 consecutive patients (49 men; median age, 59 years; range, 29-79 years) operated for acute necrotizing pancreatitis (2000-2003) in 11 hospitals were reviewed. Five abdominal radiologists independently categorized the peripancreatic collections according to the Atlanta classification. Radiologists were aware of the timing of the CT and the clinical condition of the patient. Interobserver agreement was determined. RESULTS: Interobserver agreement among the radiologists was poor (kappa, 0.144; SD, 0.095). In 3 (4%) of 70 cases, the same Atlanta definition was chosen. In 13 (19%) of 70 cases, 4 radiologists agreed, and in 42 (60%) of 70 cases, 3 radiologists agreed on the definition. In 21 cases (30%), 1 or more of the radiologists classified a collection as "pancreatic abscess," whereas 1 or more radiologist used another Atlanta definition. CONCLUSION: The interobserver agreement of the Atlanta classification for categorizing peripancreatic collections in acute pancreatitis on CT is poor. The Atlanta classification should not be used to describe complications of acute pancreatitis on CT. PMID- 17079936 TI - The incidence and case-fatality rates of acute biliary, alcoholic, and idiopathic pancreatitis in California, 1994-2001. AB - OBJECTIVE: To better define the epidemiology of acute pancreatitis in a racially diverse population. METHODS: Analysis of all patients hospitalized in California with first-time acute pancreatitis for the period between January 1994 and September 2001. Subtypes were classified based on the presence or absence of predisposing conditions. RESULTS: There were 70,231 patients hospitalized for first-time acute pancreatitis; 32.6% had biliary tract disease alone, 20.3% had alcohol abuse alone, and 36.6% were idiopathic. The age-standardized incidence increased by 32% from 33.2 to 43.8 cases per 100,000 adults for the period between 1994 and 2001, with the largest increase in the biliary group (52%). The standardized incidence rate of alcoholic and idiopathic pancreatitis was highest in African Americans, whereas biliary pancreatitis was highest in Hispanics. There was no change over time in the percentage of patients dying in the first 14 or 91 days; and in a risk-adjusted model, patients with alcoholic pancreatitis had the highest risk of dying. CONCLUSIONS: The incidence rate of acute pancreatitis rose for the period between 1994 and 2001. However, there was no reduction in the 14- or 91-day case-fatality rate. Further research is needed to explain both the rise in the incidence rate of pancreatitis and the absence of any improvement in the early case-fatality rate. PMID- 17079937 TI - Autoimmune pancreatitis can be classified into early and advanced stages. AB - OBJECTIVES: Although a number of pathological studies using various names/synonyms for autoimmune pancreatitis (AIP) have been reported, they do not mention the pathological staging related to origin/pathogenesis. Here, we propose a pathological staging for AIP lesions. METHODS: We histopathologically examined pancreatic tissue specimens of 31 AIP patients (14 pancreatectomized and 17 needle-biopsied materials) provided by 15 hospitals in Japan and studied the relevance of clinical manifestations to the pathological stage of AIP. RESULTS: Based on the presence or absence of acinar cells in AIP lesions, pancreatic tissue specimens were successfully divided into 20 cases in the early stage and 11 cases in the advanced stage, respectively. In the early stage, fibrosis was distributed in the interlobular and intralobular areas, admixed with acinar atrophy. Lymphoplasmacytic infiltration caused the narrowing of the ductal lumen and obliterative phlebitis. The common bile duct wall was also involved. In the advanced stage, the lesion was replaced by massive/extensive interlobular fibrosis with lymphoplasmacytic infiltrates to various degrees. Phlebitis was mild. Comparative analysis of clinical parameters between the early and advanced stages showed a significantly higher prevalence of jaundice and positive antinuclear antibodies in the early stage, and decreased serum lipase levels in the advanced stage. CONCLUSIONS: Autoimmune pancreatitis can be divided into early and advanced stages according to the presence or absence of acinar cells. Our pathological staging will facilitate understanding and evaluation of the clinical course in AIP. PMID- 17079938 TI - Can pancreatic steatosis explain the finding of pancreatic hyperenzymemia in subjects with dyslipidemia? AB - OBJECTIVE: It has been proposed by some investigators that benign pancreatic hyperenzymemia could result from pancreatic steatosis that they believe would have been caused by dyslipidemia; their diagnosis of steatosis was based on the finding of a hyperechogenic pancreas at ultrasound. The aim of this study was to assess the validity of this proposed model. METHODS: The study group was composed of 18 subjects with benign pancreatic hyperenzymemia, 12 men and 6 women; mean age, 55 years; range, 38 to 68 years. All 18 had dyslipidemia and 9 had hyperechogenic pancreas at ultrasound. In addition, 6 subjects with benign pancreatic hyperenzymemia but who did not have dyslipidemia or hyperechogenic pancreas and 10 healthy subjects with none of these conditions were also studied as controls. In each of these subjects, magnetic resonance imaging of the pancreas was performed to assess the presence of pancreatic steatosis. RESULTS: Magnetic resonance imaging showed normal pancreas with no signs of fatty infiltration in all 18 subjects with dyslipidemia, including those with both dyslipidemia and hyperechogenic pancreas at ultrasound. A similar result was found in all control subjects. CONCLUSION: The finding of a completely normal pancreas at magnetic resonance imaging does not support the proposed model in which pancreatic hyperenzymemia in subjects with dyslipidemia is attributed to pancreatic steatosis. PMID- 17079939 TI - Frey procedure in the treatment of chronic pancreatitis: short-term results. AB - OBJECTIVE: The aim of this multicenter study was to report the short-term results of the Frey procedure in the treatment of chronic pancreatitis. METHODS: For the period between September 2000 and January 2005, 34 Frey procedures were performed for chronic pancreatitis in 4 university hospitals. This study includes 31 men (91%) and 3 women (9%), with a mean age of 48 +/- 6 years (range, 32-58 years). The etiology of chronic pancreatitis was chronic alcohol ingestion in 32 patients (94%) and hereditary chronic pancreatitis in 2 patients. The indications of surgery were abdominal pain in all patients, requiring opiates in 59% (n = 20) and associated with a weight loss in 79% (n = 27). RESULTS: There was no mortality. Eleven postoperative surgical complications occurred in 7 patients (20%). Three patients had a single complication, and 4 patients had 2 complications. Pancreatic fistula occurred in 4 patients and healed under conservative management in all cases. One patient had massive bleeding from the stump of gastroduodenal artery requiring reoperation. The mean hospital stay was 16 +/- 8 days (range, 9-40 days). The mean follow-up was 15 +/- 12 months (range, 3-37 months). At the time of the last follow-up visit, the examiner judged that 19 patients (56%) have complete pain relief and 11 patients (32%) have substantial pain relief. No patient used narcotic analgesics postoperatively. Seven patients developed diabetes mellitus, requiring insulin (n = 1), oral hypoglycemic agents (n = 5), and diet adjustment (n = 1). Four patients developed exocrine insufficiency. Weight increases with a mean of 4.8 +/- 5.4 kg (range, 1 24 kg) in 27 patients (79%). CONCLUSIONS: Frey procedure appears as a safe technique with low mortality and morbidity rates and allows effective pain relief in about 90% of patients. PMID- 17079940 TI - Significant increase of serum high-mobility group box chromosomal protein 1 levels in patients with severe acute pancreatitis. AB - OBJECTIVE: Multiple organ failure because of systemic inflammatory response in the early phase and sepsis in the late phase is the main contributor to high mortality in severe acute pancreatitis (SAP). High-mobility group box chromosomal protein 1 (HMGB1) was recently identified as a potent proinflammatory mediator and increases in various pathological conditions such as sepsis. The aim of this study was to investigate contributions of HMGB1 in SAP. METHODS: We measured serum HMGB1 concentrations by an enzyme-linked immunosorbent assay in 45 patients with SAP at the time of admission. Furthermore, relationship between their serum HMGB1 levels and clinical factors was analyzed. RESULTS: The mean value of serum HMGB1 levels was significantly higher in patients with SAP (5.4 +/- 1.3 ng/mL) than that in healthy volunteers (1.7 +/- 0.3 ng/mL). Serum HMGB1 levels were significantly positively correlated with the Japanese severity score and Glasgow score. Serum HMGB1 levels were significantly positively correlated with lactate dehydrogenase, C-reactive protein, and total bilirubin. The HMGB1 levels were higher in patients with organ dysfunction and infection during the clinical course. The HMGB1 levels in nonsurvivors were higher than those in survivors. Serum HMGB1 levels gradually declined after the admission. CONCLUSIONS: Serum HMGB1 levels were significantly increased in patients with SAP and were correlated with disease severity. These results suggest that HMGB1 may act as a key mediator for inflammation and organ failure in SAP. PMID- 17079941 TI - Cigarette smoke-induced differential expression of the genes involved in exocrine function of the rat pancreas. AB - OBJECTIVE: Little is known about the molecular and biological aspects of the epidemiological association between smoking and pancreatic pathology, such as chronic pancreatitis and pancreatic cancer. Recently, we reported that tobacco smoke exposure induced morphological alterations in the rat pancreas. Here, we have investigated the alterations in the expression of genes associated with exocrine pancreatic function and cellular differentiation upon exposure to cigarette smoke. METHODS: Female rats were exposed to environmental smoke inhalation for 2 d/wk (70 min/d) for 12 weeks. The expression profiles of trypsinogen, pancreas-specific trypsin inhibitor, cholecystokinin A receptor, cystic fibrosis transmembrane conductance regulator (CFTR), carbonic anhydrase, and Muc1 and Muc4 mucins transcripts were analyzed by RNA slot blot analysis. Muc4 expression was also examined by immunohistochemistry. RESULTS: Our data revealed that the ratio of trypsinogen to that of the protective pancreas specific trypsin inhibitor was elevated upon cigarette smoke exposure. The expression of carbonic anhydrase and CFTR remained unaltered when inflammatory signs were not detected in histological examinations. On the other hand, when pancreatic inflammation was present, the levels of CFTR and carbonic anhydrase were increased, indicating ductal and/or centroacinar cell involvement. No changes in the expression of Muc1 and Muc4 mucins were observed. CONCLUSIONS: Our data show that cigarette smoke exposure leads to an increased vulnerability to pancreatic self-digestion. Moreover, the concomitant involvement of pancreatic ducts occurs only when focal pancreatic inflammation is present. PMID- 17079942 TI - Establishment of animal models for three types of pancreatitis and analyses of regeneration mechanisms. AB - OBJECTIVES: To investigate the mechanisms underlying the onset and progress of pancreatitis, 3 animal models (chronic, acute, and severe pancreatitis) were established by double ligature of the pancreatic duct, injection with cerulein, or injection with cerulein + double ligature of the pancreatic duct. METHODS: We prepared a control and 3 experimental groups: group 1 (untreated control), group 2 (a chronic pancreatitis model; the pancreatic tail was exposed through a midline incision, and the pancreatic duct from this part was double-ligated), group 3 (an acute pancreatitis model; cerulein was intraperitoneally injected 7 times on day 0), and group 4 (a severe pancreatitis model; the double ligature of the pancreatic duct plus injection of cerulein). RESULTS: Kinetic observations of survival rate, relative pancreatic weight, and the macroscopical and microscopical diagnoses and observations of the changes in endocrine function clearly show that these 3 murine models of pancreatitis can serve as human models for chronic, acute, and severe pancreatitis. Furthermore, pancreas duodenum homeobox 1, cytokeratin 19, and Ki67 are expressed at the site of injury in the pancreas, resulting from the injection with cerulein and/or double ligature of the pancreatic ducts and indicating that there remains a tissue-regenerative capacity. CONCLUSIONS: These 3 mouse models could serve as human models for chronic, acute, and severe pancreatitis. Furthermore, cells of the epithelial lineage might participate in tissue regeneration in chronic, acute, and severe pancreatitis. PMID- 17079943 TI - Clinical characteristics of pancreatic neuroendocrine tumors in Japanese patients with von Hippel-Lindau disease. AB - OBJECTIVE: The aim of this study was to elucidate the clinical characteristics of pancreatic neuroendocrine tumors (NET) in Japanese patients with von Hippel Lindau (VHL) disease. METHODS: We sent a questionnaire to all members of the Japan Pancreas Society in 2002 asking for the number of patients with VHL and complications of NET and/or cystic lesion in the pancreas. Furthermore, we sent a second questionnaire to obtain detailed information about the clinical characteristics of pancreatic NET. RESULTS: A total of 58 patients with VHL were reported. Among these, 34 (59%) patients had pancreatic lesions, including 10 with pancreatic NET, 23 with a cystic lesion, and one with both. The mean age at identification of pancreatic NET was 34.6 years (range, 22-64 years). The mean diameter of the tumors was 4.3 cm (range, 1-12.5 cm). Distant metastases were found in 2 (20%) cases. During the follow-up period (3.3 years; range, 0-8 years), 7 patients are alive, and 2 patients died of hemangioblastomas. CONCLUSIONS: Pancreatic NET in the VHL disease showed a relatively lower incidence of metastasis compared with sporadic non-functioning pancreatic NET, yielding a favorable prognosis. Because they present no hormonal syndrome, periodic screening examinations are warranted to identify pancreatic NET at an early stage. PMID- 17079944 TI - Clinical roles of increased populations of Foxp3+CD4+ T cells in peripheral blood from advanced pancreatic cancer patients. AB - OBJECTIVES: Further metastasis should be avoided in pancreatic cancer (PC) patients for effective surgical treatment. Regulatory T cells (Foxp3CD4 T cells including CD4CD25 T cells and CD4CD25 T cells) play important roles in tumor immunity. This study aimed to investigate whether regulatory T cells participate in metastasis. METHODS: Peripheral blood was withdrawn from PC patients, as well as healthy volunteer donors as controls. The peripheral blood mononuclear cells (PBMCs) were subjected to FACScan analysis after labeling with anti-CD4, anti CD25, and anti-Foxp3 antibodies. Tumor markers, including DUPAN2 and CA19-9, surface markers, such as the CD4/CD8 ratio, and the CD57 cell population were assessed. Clinical stages were classified according to the TNM classification. RESULTS: The Foxp3CD4 T-cell population among the PBMCs was significantly increased in PC patients (8.10% +/- 4.65%) compared with healthy donors (2.47 +/- 0.78%) (P < 0.001). No significant relationships existed for the tumor markers, CD4/CD8 ratio, and CD57 cells. However, a significant correlation was found between Foxp3CD4 T cells among the PBMCs and the TNM stage (P < 0.05). CONCLUSIONS: Foxp3CD4 T cells are good markers for metastasis detection in PC patients and more accurate than other conventional tumor markers, especially at advanced stages of the disease. PMID- 17079945 TI - The expression of MUC4 and MUC5AC is related to the biologic malignancy of intraductal papillary mucinous neoplasms of the pancreas. AB - OBJECTIVE: Intraductal papillary mucinous neoplasms (IPMNs) of the pancreas show heterogeneous proliferations with latent malignancy. Mucins (MUC) are high molecular-weight glycoproteins, with an aberrant expression profile in various malignancies. Recently, MUC4 and MUC5AC expressions have been demonstrated to correlate with the unfavorable and the favorable prognosis of pancreatic duct cell carcinoma, respectively. However, little is known about these mucin expressions in IPMNs. METHODS: To clarify the role of MUC4 and MUC5AC expressions in IPMNs, the expression profiles of MUC4 and MUC5AC were investigated in 50 lesions from 17 specimens with 16 IPMNs by immunohistochemistry, using each of their specific antibodies. RESULTS: The expression of MUC4 was found in the lesions ranging from adenoma to cancer lesions of IPMNs, whereas it was undetectable in normal and hyperplastic lesions. Frequent expression of MUC4 is found in the higher grade of IPMNs (borderline and cancer lesions; 16/18 lesions, 94%). The differences were independently significant (P < 0.001) when the cutoff point was set between adenoma and borderline IPMNs. Similarly, frequent expression of MUC5AC was detected in the lesions from adenoma to cancer of IPMNs (32/34, 94%), whereas no intense expression was detected in normal or hyperplastic lesions. The significant difference was found when the cutoff point was set between hyperplasia and adenoma of IPMNs (P < 0.001). CONCLUSIONS: These results indicated that the expressions of MUC4 and MUC5AC are potential markers to distinguish adenoma or above malignant lesions of IPMNs from lesser malignant ones, respectively. PMID- 17079946 TI - Gemcitabine chemotherapy versus 5-fluorouracil-based concurrent chemoradiotherapy in locally advanced unresectable pancreatic cancer. AB - OBJECTIVES: The aim of this study was to compare the survival benefits associated with gemcitabine chemotherapy and 5-fluorouracil (5-FU)-based concurrent chemoradiotherapy (CCRT) in locally advanced unresectable pancreatic cancer. METHODS: One hundred and thirty-eight locally advanced unresectable pancreatic cancer patients were retrospectively enrolled from January 1995 to January 2005. All cases were histologically proven, and patients received gemcitabine chemotherapy, 5-FU-based CCRT, or supportive care at Seoul National University Hospital. RESULTS: Median overall survival was 8.2 months. Twenty-six patients received gemcitabine chemotherapy, 56 patients 5-FU-based CCRT, and 56 patients supportive care. Weight loss and treatment modality were identified as independent prognostic factors by multivariate analysis. Patients in the 5-FU based CCRT (overall survival, 10.4 months) and gemcitabine chemotherapy (11.3 months) groups showed survival benefit over those received supportive care (6.1 months, P < 0.0001). No grades 3 to 4 toxic adverse effects occurred in either treatment group and no statistical significant survival difference was found between gemcitabine chemotherapy and 5-FU-based CCRT (P = 0.5). CONCLUSIONS: Patients with locally advanced pancreatic cancer who received gemcitabine chemotherapy or 5-FU-based CCRT showed better survival than those who received supportive care only. Gemcitabine chemotherapy and 5-FU-based CCRT showed similar survival advantages. PMID- 17079947 TI - The association of acetyl-l-carnitine and nicotinamide remits the experimental diabetes in mice by multiple low-dose streptozotocin. AB - OBJECTIVES: We studied the effect of acetyl-l-carnitine plus nicotinamide (AC + N) on murine diabetes mellitus induced by multiple low doses of streptozotocin. METHODS: Male C57BL/6J inbred mice were injected intraperitoneally with citrate buffer or streptozotocin (40 mg/kg) for 5 consecutive days, followed by injections of saline solution or AC + N (50 + 25 mg/kg) from days 6 to 110. Four groups were studied: normal control mice (C), treated normal control mice (TC), diabetic mice (D), and treated diabetic mice (TD). TD group was divided into 2 at day 86; treatment was suspended in one group (TDs) and continued in the other until day 110. RESULTS: Weight, plasma glucose, plasma insulin, cellular immune aggression, glucose-stimulated insulin secretion from perifused pancreatic slices, and pancreas histology were studied in each experimental group. Diabetic mice treated with AC + N showed improvements in weight, plasma glucose, and plasma insulin levels without mortality, reaching control values at day 110. Cellular immune aggression and insulin release from pancreatic slices perfusions improved without reaching control values. Histology showed that insulin immunostained area, the index of insulin immunostained beta cells and beta-cell size, was normalized at the end of the study. CONCLUSIONS: The treatment with AC + N induced remission of autoimmune type 1 diabetes in mice produced by multiple low doses of streptozotocin. PMID- 17079948 TI - Is defective pancreatic beta-cell mass environmentally programmed in Goto Kakizaki rat model of type 2 diabetes?: insights from crossbreeding studies during suckling period. AB - OBJECTIVE: The Goto-Kakizaki (GK) rat is a spontaneous model of type 2 diabetes with a well established pathological pancreatic beta-cell development. Hyperglycemia experienced during early postnatal life contributes to the programming of endocrine pancreas. We have analyzed the consequences of hyperglycemic versus euglycemic suckling period for the pancreatic beta-cell mass and the in vivo glucose tolerance and insulin secretion in 4-week-old unweaned control Wistar (W), diabetic GK, and in offspring issued from crosses between normoglycemic W and diabetic GK rats. METHODS: Mother/father crosses yielded offspring designated as follows: W/W, GK/GK, W/GK, and GK/W. In vivo glucose tolerance and insulin secretion tests were performed on males 4 weeks after birth, that is, just before weaning. Beta-cell mass was determined by immunohistochemistry and morphometry. RESULTS: Four-week-old W/GK and GK/W rats are normoglycemic, normoinsulinemic, and display a similarly small beta-cell mass. Both W/GK and GK/W rats exhibit in vivo glucose intolerance and defective insulin secretion in response to glucose. CONCLUSIONS: Our data obtained from crossbreeding studies during suckling period suggest that the defective pancreatic beta-cell mass is not environmentally programmed in the GK model of type 2 diabetes. Rather, they support the hypothesis that the beta-cell mass defect in the GK is linked to genetic determinism. PMID- 17079949 TI - Enalapril increases the local extravasation of macromolecules and nitric oxide synthase in pancreas of the fructose-fed insulin-resistant rat model. AB - OBJECTIVES: Angiotensin-converting enzyme (ACE) inhibitors have been associated with an increased risk of acute pancreatitis. The pathogenesis of this condition remains unclear, but an activation of the kinin system and a resultant localized angioedema have been implicated in the initial step leading to acute pancreatic damage. The goal of the present study was to explore the impact of ACE inhibition on pancreatic microcirculation and capillary permeability in normal and insulin resistant rats. METHODS: Chow- or fructose-fed Sprague-Dawley rats were treated with enalapril (dosage, 10 mg.kg.d) or vehicle for 4 weeks before measuring in vivo the extravasation of Evans blue (EB) dye in pancreas. Unanesthetized animals (n = 10-17 per group) were injected with EB 20 mg.kg in the caudal vein 10 minutes before killing, and EB dye was extracted from each pancreas by using formamide. RESULTS: Relative to controls, enalapril-treated animals showed a 5 fold increase in pancreatic extravasation of EB in the fructose-fed rat model (P < 0.001); smaller changes (2-fold) were observed in the chow-fed animals treated with enalapril (P < 0.001). The increase in pancreatic vasopermeability observed with enalapril in the fructose-fed animals was accompanied by a significant increase in total pancreatic nitric oxide synthase (NOS) activity compared to controls (Delta = +128%; P < 0.001). This increase in NOS activity seemed to be solely attributable to an upregulation of the endothelial NOS isoform because only the eNOS immunoreactive mass (as opposed to nNOS) seemed to be increased in the pancreas of these animals. Treatment with enalapril was not associated with any increase in serum amylase concentrations in either animal subgroup. CONCLUSIONS: Enalapril increases capillary permeability (extravasation of macromolecules) in the pancreas of the fructose-fed rat model. This suggests that ACE inhibition upregulates the eNOS isoform locally, increases vasopermeability of the pancreas, and can therefore result in local edema in the fructose-fed insulin-resistant rat model. PMID- 17079950 TI - Pancreatic abscess caused by Corynebacterium coyleae mimicking malignant neoplasm. AB - A 50-year-old female was referred to our hospital because of postprandial epigastric pain and pancreatic head mass. On admission, an elastic hard mass with tenderness was palpable in the epigastric region. Laboratory findings showed no abnormalities, except for a slightly elevated C-reactive protein value and iron deficiency anemia. Serum levels of pancreatic enzymes and tumor markers were also within the reference range. Computed tomography (CT) demonstrated a 5-cm heterogenous mass at the head of the pancreas. Angiography showed that gastroduodenal artery was transformed and narrowed by the mass. Smooth stenosis of portal vein was also observed. Fusion CT-positron emission tomography with 2 deoxy-2-[F]fluoro-D-glucose demonstrated a focus of increased uptake in the pancreatic head mass. We suspected the mass of malignancy but, surprisingly, tumor size was gradually decreased without any therapies. Biopsy specimens from the mass of the pancreas showed marked inflammatory cell infiltration and marked interstitial fibrosis without malignant cells. Thereafter, we could isolate Corynebacterium coyleae from the biopsy specimen. We diagnosed the mass as a pancreatic abscess caused by C. coyleae and started with the intravenous antibiotics therapy. Subsequent follow-up CT and ultrasonography showed dramatic improvement in pancreatic mass. We present here a case of pancreatic abscess which was difficult to differentiate from malignant lesion by various imaging studies. Moreover, we could culture and identify C. coyleae which had never been reported to be the source of pancreatic abscess. PMID- 17079951 TI - Pancreatic cancer in cystic fibrosis after bilateral lung transplantation. AB - Cystic fibrosis (CF) is the most common lethal genetic disease affecting 1 in 2500 white patients. Chronic obstructive lung disease and pancreatic insufficiency are the main clinical manifestations of CF. Lung transplantation has become a treatment option for advanced pulmonary disease during the last decade. On the other hand, there is evidence from large cohort studies that CF and immunosuppression are risk factors for pancreatic cancer. Here, we report the case of an 18-year-old female patient with CF and bilateral lung transplantation who underwent Whipple procedure for pancreatic adenocarcinoma at the age of 12 years. Because of growth retardation, the patient underwent a 2-year period of growth hormone treatment before the diagnosis of pancreatic cancer. This case should sensitize physicians to be aware of the increased risk for pancreatic cancer in CF patients especially in those after lung transplantation with immunosuppression and prolonged survival. PMID- 17079952 TI - Long-term maintenance of graft function after islet autotransplantation of less than 1000 IEQ/kg. PMID- 17079953 TI - Successful treatment of advanced-stage autoimmune pancreatitis-related sclerosing cholangitis. PMID- 17079954 TI - Adenocarcinoma of the pancreas associated with dorsal agenesis. PMID- 17079957 TI - Primary lateral sclerosis with MR diffusion tensor image and tract tracking. PMID- 17079958 TI - Sensory neurons and fibers from multiple spinal cord levels innervate the rabbit lumbar disc. AB - OBJECTIVE: To establish the neurotransmission pathway from the lumbar L5/6 intervertebral disc (IVD) to the spinal cord in the rabbit. DESIGN: Fluorogold particles injected into the posterior portion of the rabbit L5/6 IVD were traced by examining gold-positive neurons and fibers in the dorsal root ganglion (DRG) and spinal cord at various root levels. RESULTS: Fluorogold-labeled neurons were observed bilaterally in primary afferent DRG neurons from the L3 through L5 segments; a small number of gold-labeled neurons were found at the L1 level. Fluorogold-labeled neurons were predominantly present in the ipsilateral DRG (the side of the injection) at the L5 level, but they were more equally distributed (on both sides) at the L4 and L3 levels. In the posterior horn of the spinal cord, Fluorogold particles were found in nerve fibers as rostral as the T12 level. CONCLUSIONS: Our study has shown that Fluorogold particles injected into the rabbit L5/6 IVD are taken up by primary sensory neurons in the DRGs and primary sensory fibers in the posterior horn of the spinal cord at multiple levels. This diffuse innervation pattern of the lumbar disc may help explain why discogenic back pain in humans is often poorly localized. PMID- 17079959 TI - A preliminary examination of the CMS eligibility criteria in total-joint arthroplasty. AB - OBJECTIVE: To analyze inpatient rehabilitation outcomes in total-knee arthroplasty (TKA) and total-hip arthroplasty (THA) patients using the 2004 Medicare 75% rule criteria. DESIGN: This retrospective study compared outcomes in unilateral TKA (UTKA), bilateral TKA (BTKA), and THA after interdisciplinary inpatient rehabilitation (n = 867). Patients were separated into three comparison pairs: 1) UTKA or BTKA, 2) age <85 yrs or > or =85 yrs, and 3) body mass index (BMI) <50 or > or =50 kg/m. Length of stay (LOS), functional independence measure (FIM) scores (total, motor, and cognitive), hospital charges, FIM efficiency, and discharge disposition were analyzed. RESULTS: BTKA improved total FIM score more than UTKA (43 vs. 38%; P = 0.039). TKA with BMI > or =50 kg/m had similar admission and discharge FIM motor scores compared with BMI <50 kg/m (P > 0.05). TKA patients > or =85 yrs had lower admission FIM scores, longer LOS (11.3 vs. 9.4 days), and 22% higher total charges than TKA patients younger than 85 yrs (P = 0.042). THA patients > or =85 yrs had 6-10% lower total FIM, FIM motor, and FIM cognition scores and were discharged to home less frequently than younger patients (P < 0.05). Total and daily charges were 21-162% higher in THA patients with BMIs > or =50 kg/m than in THA patients with BMIs <50 kg/m (P < 0.045). CONCLUSION: All patients made functional gains during rehabilitation. However, the most costly and lengthy rehabilitation occurred in TKA patients > or =85 yrs and THA patients with BMI > or =50 kg/m. PMID- 17079960 TI - Outcomes of a consult fall prevention screening clinic. AB - OBJECTIVE: To report patient satisfaction and quality outcomes from a standardized, multicenter fall prevention screening clinic. DESIGN: Referrals were made to the fall prevention screening clinic, which was staffed by a physician, pharmacist, therapist, nurse, and the site coordinator. The fall prevention screening clinic staff utilized a team-meeting format with clearly delineated management options to determine the appropriate interventions based on various screening examinations. RESULTS: Primary care providers acknowledged pharmacy recommendations and >80% of the recommendations were followed. There was a reduction in reported falls (initial visit = 297 falls, follow-up visit = 141 falls, P = 0.0002) and in the number of recurrent fallers. There was a small (average difference = 1.8 secs) but significant (P = 0.025) decrease in the 8 foot Up-and-Go time, representing a functional improvement in walking speed. There was no significant change in the Fall Efficacy Scale, however. At least 80% of the patients reported that they were completely satisfied with their care. CONCLUSIONS: Patients who attended the fall prevention screening clinic demonstrated improved confidence during activities of daily living and reduced falls. They reported high satisfaction with the clinic format. PMID- 17079961 TI - Life satisfaction after traumatic brain injury and the World Health Organization model of disability. AB - OBJECTIVE: To determine which components of the World Health Organization's International Classification of Functioning, Disability, and Health (ICF) are most predictive of global life satisfaction after traumatic brain injury (TBI). DESIGN: Prospective evaluation of 180 individuals enrolled in a TBI model system project site. RESULTS: Multiple regression analysis indicated that the combination of ICF components (body function and structure, activities, and participation) and demographic factors significantly predicted life satisfaction and accounted for 17% of the variance. Participation was the strongest predictor; activities were a significant, but weaker predictor; and body function and structure did not add to the prediction of life satisfaction. Of all the individual variables evaluated, only social integration and productivity were found to be significant, unique predictors. CONCLUSIONS: When considering the effects of various aspects of disabling conditions on the life satisfaction of individuals who have suffered a TBI, restriction of participation in life activities was found to have the greatest impact. Although the model accounted for a significant percentage in the variation of life satisfaction, a large proportion of the predictive picture (>82%) remains unclear. Doubtless, other variables impinge on life satisfaction that would further clarify the complex relationship between disabling conditions and life satisfaction in TBI. PMID- 17079962 TI - The effect of rolling resistance on stationary wheelchair wheelies. AB - OBJECTIVE: To test the hypotheses that increased rolling resistance (RR) reduces rear-wheel displacement and perceived difficulty during the takeoff and balance phases of stationary wheelchair wheelies. DESIGN: We carried out within-subject comparisons of 20 participants as they each performed, in random order, two 30 sec stationary wheelies in three RR settings (tile, 5-cm-thick foam, and 12.5-cm high blocks in front of and behind the rear wheels). The main outcome measures were rear-wheel displacement (in centimeters for the takeoff phase and centimeters per second for the balance phase) from a spring-loaded potentiometer and Likert scales of perceived difficulty. RESULTS: For rear-wheel displacement, all six of the pairwise comparisons (three terrains x two phases (takeoff and balance)) showed a significant statistical difference (P < 0.002). In each of the six pairwise comparisons, displacement was less for the higher of the two RR conditions. For perceived difficulty, during the balance phase, participants perceived tile to be significantly more difficult than either foam (P = 0.0067) or blocks (P = 0.0002). The other pairwise comparisons were not statistically significant. CONCLUSION: In conditions of increased RR, rear-wheel displacement and perceived difficulty are reduced during stationary wheelchair wheelies. These findings have implications for teaching wheelchair users to perform wheelies, a foundation of many advanced wheelchair skills. PMID- 17079963 TI - Specificity of needle electromyography for lumbar radiculopathy and plexopathy in 55- to 79-year-old asymptomatic subjects. AB - OBJECTIVE: Determine specificity of needle electromyography for lumbar radiculopathy and plexopathy using a blinded study design. DESIGN: Asymptomatic community volunteers ages 55 and older, as part of a spinal stenosis study, were given a standardized electrodiagnostic evaluation by a blinded electromyographer. A monopolar needle was used to evaluate five leg muscles and the lumbar paraspinal muscles. The specificities of different diagnostic criteria for radiculopathy and plexopathy were then calculated. RESULTS: There were 30 subjects with a mean age of 65.4 yrs (SD 8.0). When only positive sharp waves or fibrillations were counted as abnormal, most of the diagnostic criteria (two limb muscles plus associated lumbar paraspinal muscle abnormal, two limb muscles abnormal, or one limb muscle plus associated lumbar paraspinal muscle abnormal) had 100% specificity. When we also included at least 30% polyphasia in the limb muscles as abnormal, the respective specificities were 97, 90, and 87%. When we also included at least 20% polyphasia in the limb muscles as abnormal, the respective specificities were 77, 60, and 60%. The specificity for plexopathy was 100% when only positive sharp waves or fibrillations were used, and it remained 100% when increased polyphasia was added. CONCLUSION: Needle electromyography has excellent specificity for lumbosacral radiculopathy and plexopathy when appropriate diagnostic criteria are used. PMID- 17079965 TI - Vitamin D deficiency: implications in the rehabilitation setting. AB - OBJECTIVE: Vitamin D deficiency, which can result from inadequate sun exposure, dietary intake, or problems with absorption, is rarely documented in the rehabilitation literature. Most likely, it is rarely thought of by the rehabilitation profession. This is problematic because vitamin D deficiency can present as musculoskeletal pain, which is commonly seen in both outpatient clinics and inpatient rehabilitation units. The populations with the greatest risk include the homebound elderly, people with pigmented skin, people with cultural and social avoidance of the sun, people who live in wintertime in climates above and below latitudes of 35 degrees, and people with gastrointestinal malabsorption. DESIGN: The review was done using PubMed, Ovid, and MDConsult using the search terms pain, chronic pain, musculoskeletal pain, vitamin D deficiency, and osetomalacia. The search revealed 107 articles and was narrowed down to 51 articles that focused on vitamin D deficiency and its musculoskeletal manifestations. RESULTS: A direct correlation was noted between vitamin D deficiency and musculoskeletal pain. At-risk populations are not acquiring enough vitamin D through sun exposure, and the current recommended daily allowances from dietary sources including supplements are too low to compensate for this lack of sun exposure. Treatment of vitamin D deficiency produced an increase in muscle strength and a marked decrease in back and lower limb pain within 6 mos. CONCLUSION: Vitamin D deficiency should be included in the differential diagnosis in the evaluation of musculoskeletal pain complaints in the rehabilitation setting, and treatment of any identified deficiency should be considered a potentially important component of the treatment regimen. PMID- 17079966 TI - Acute arterial occlusion after total knee arthroplasty. AB - There are a number of complications associated with total knee-joint arthroplasty. These include deep venous thromboses, peroneal palsy, infection, anemia, and Ogilvie's syndrome. An uncommon but potentially limb-threatening complication is acute arterial occlusion. Approximately 35 cases have been reported in the orthopedic literature. Prompt recognition and treatment intervention are the keys to successful outcome. We describe the case of one patient who had mild peroneal palsy and developed acute arterial occlusion 9 days postoperatively while on the inpatient rehabilitation service. Prompt aggressive management restored arterial circulation to the lower limb. Careful management of patients after total knee arthroplasty requires an understanding that arterial occlusion is a rare limb-threatening complication of surgery, but that it is treatable with prompt, deliberate management. Physiatrists should be aware that this condition exists in postoperative knee-joint arthroplasty patients. They should pay careful attention to any patient with a history of peripheral vascular disease or postoperative peroneal palsy. PMID- 17079967 TI - Hand function improvement with low-frequency repetitive transcranial magnetic stimulation of the unaffected hemisphere in a severe case of stroke. AB - Previous research has shown that low-frequency rTMS of the unaffected hemisphere can improve motor function in acute and chronic stroke patients. However, these studies only investigated patients with mild or moderate motor deficits. We report a case of a stroke patient with a severe motor impairment who underwent sham and active repetitive transcranial magnetic stimulation (rTMS) of the unaffected hemisphere and had significantly improved motor function after active, but not after sham, stimulation of the unaffected primary motor cortex. In an additional session of active rTMS, this patient maintained and further enhanced the initial motor improvement. This case report shows that inhibitory rTMS of the unaffected hemisphere can also be beneficial for stroke patients with severe motor deficits and suggests that this approach of noninvasive brain stimulation should be further investigated in this population of patients. PMID- 17079968 TI - Wheeling a petit pas: Parkinsonism detected by observation of wheelchair propulsion. AB - We present a man with parkinsonism detected by the observation of wheelchair propulsion. His manual wheelchair propulsion technique was observed to include rapid, brief, low-power strokes resembling the marche a petit pas (walking with tiny steps) phenomenon of parkinsonism. We videotaped his wheelchair propulsion and compared him with ten age-, gender-, and diagnosis-matched controls. The patient had a propulsion velocity of 0.14 m/sec compared with a mean (+/- standard deviation) of 0.73 (+/- 0.16) m/sec for the controls, a cadence of 209 strokes/min vs. 60 (+/- 12) strokes/min for the controls, and a mechanical efficiency of 0.04 m/stroke compared with 0.75 (+/- 0.25) m/stroke for the controls. This observation shifted the course of his medical investigations and management as well as his rehabilitation care. This is the first detailed report of how parkinsonian features may affect manual wheelchair propulsion. It suggests that observation of wheelchair mobility should be a routine component of the physical examination of wheelchair users. PMID- 17079970 TI - What is the American physiatrist's role in the International Physical Medicine and Rehabilitation Organization? PMID- 17079975 TI - Pharmacotherapy of osteoarthritis. PMID- 17079976 TI - Osteoarthritis: epidemiology, risk factors, and pathophysiology. PMID- 17079978 TI - Viscosupplementation (biosupplementation) for osteoarthritis. PMID- 17079979 TI - Corticosteroid injections for osteoarthritis. PMID- 17079981 TI - Osteoarthritis and therapeutic exercise. PMID- 17079983 TI - Orthotics and assistive devices in the treatment of upper and lower limb osteoarthritis: an update. PMID- 17079984 TI - Rehabilitation after hip- and knee-joint replacement. An experience- and evidence based approach to care. PMID- 17079988 TI - Total tendon transplant for slipping patella: a new operation for recurrent dislocation of the patella. 1938. PMID- 17079989 TI - Recurrent dislocation of the patella: operative treatment. 1888. PMID- 17079990 TI - Wound problems in total knee arthroplasty. AB - Wound problems can often be prevented with careful planning. When transverse incisions are used for knee surgery many years prior to any anticipated knee arthroplasty, no major problems are typically encountered with a conventional, anterior longitudinal incision. We recommend lateral incisions (eg, after a previous lateral tibial plateau fracture) be reused for TKA. When confronted with multiple previous incisions, surgeons would best use the most recently healed or the most lateral. We prefer soft tissue reconstruction with expanders or a gastrocnemius flap if there are multiple incisions, if the skin and scar tissue are adherent to underlying tissue, or if wound healing seems questionable. Deep infection must be determined by aspiration. When present, we believe treatment must include irrigation, debridement, polyethylene exchange if acute, and resection arthroplasty if chronic. Poor wound healing is a potentially devastating complication that may result in multiple reconstructive procedures and even amputation. Early recognition followed by expeditious debridement and soft tissue reconstruction should be used for managing wound complications after TKA. PMID- 17079992 TI - Pharmacokinetics and safety of tenofovir disoproxil fumarate on coadministration with lopinavir/ritonavir. AB - OBJECTIVE: Lopinavir/ritonavir (LPV/r) and tenofovir disoproxil fumarate (TDF) are frequently used antiretrovirals. A pharmacokinetic study in healthy volunteers was conducted to assess the potential for a drug interaction between these agents. METHODS: This was a 36-day, multiple-dose, drug-drug interaction study of TDF and lopinavir/ritonavir (LPV/r). Subjects received TDF alone for 7 days, followed by 14 days each of TDF plus LPV/r and LPV/r alone in a randomized manner. Pharmacokinetic assessments were performed over 24 hours on days 7, 21, and 35. LPV/r and tenofovir plasma/serum concentrations were measured by high performance liquid chromatography/mass spectometry (MS)/MS. Geometric mean ratios and 90% confidence intervals of pharmacokinetic parameters for tenofovir, LPV, and ritonavir (RTV) were estimated using analysis of variance and compared with the no-effect criterion for pharmacokinetic equivalence. RESULTS: Tenofovir measurements with an area under the concentration-time curve over the dosing interval, maximum concentration, and concentration at the end of the dosing interval (Ctau) were 32%, 15%, and 51% higher, respectively, when TDF was coadministered with LPV/r (n = 24). LPV and RTV pharmacokinetics, including Ctau, were unaffected by TDF (n = 24). Clinical estimates of renal function were unaffected by administration of TDF alone or with LPV/r. DISCUSSION: Coadministration of TDF with LPV/r resulted in increased tenofovir exposures at steady state, possibly through increased absorption. This increase is not believed to be clinically relevant based on the safety and efficacy of TDF plus LPV/r-containing regimens in HIV-infected patients in long-term controlled clinical trials. PMID- 17079993 TI - Contribution of bacterial sepsis to morbidity in infants born to HIV-infected Haitian mothers. AB - BACKGROUND: Haiti is a country with a heavy burden of HIV infection in childbearing women. Previous studies have shown that early infant deaths are common in children of HIV-infected women. This study was designed to define the rates of and risk factors for systemic bacterial and mycobacterial infection in such children and to identify the causative agents. METHODS: A cohort of 120 children born to HIV-infected mothers between May 2001 and December 2003 were prospectively observed to 15 months of age. They received comprehensive pediatric care at the GHESKIO Centers. Children were assigned to being HIV-infected by serology, RNA detection, and/or defining clinical illnesses. Blood cultures were obtained before giving antibiotics in children who were febrile or chronically ill. Blood cultures also were obtained at selected visits on well children. RESULTS: The mortality rate in the first 15 months was high, 22 of 106 (207/1,000 live births) in these children. Sixteen (70%) deaths were within 6 months of birth. Fourty-eight blood cultures had clinically significant organisms of which 38 were Staphylococcus aureus. Blood cultures were more likely to be positive in symptomatic and in HIV-infected children. CONCLUSIONS: Despite perinatal HIV treatment, mortality in children born to HIV-infected mothers remained high. Bacteremia, particularly with Staphylococcus aureus, is a partial explanation for excess illness. PMID- 17079994 TI - CD4 T-cell count and HIV-1 infection in adults with uncomplicated malaria. AB - BACKGROUND: HIV-1-negative children with malaria have reversible lymphocyte and CD4 count decreases. We assessed the impact of malaria parasitemia on the absolute CD4 count in both HIV-1-infected and non-HIV-infected adults. METHODS: In Ndola, Zambia, at the health-center level, we treated 327 nonpregnant adults for confirmed, uncomplicated, clinical malaria. We assessed HIV-1 status, CD4 count, and HIV-1 viral load (if HIV-1-infected) at enrollment and at 28 and 45 days after treatment. RESULTS: After successful antimalarial treatment, the median CD4 count at day 28 of follow-up increased from 468 to 811 cells/microL in HIV-1-negative and from 297 to 447 cells/microL in HIV-1-positive patients (paired t test, P < 0.001 for both). CD4 count increment was inversely correlated with CD4 count at day 0 in both HIV-1-negative (P < 0.001) and HIV-1-positive patients (P = 0.03). After successful treatment, the proportion of patients with CD4 count <200/microL at day 45 decreased from 9.6% to 0% in HIV-1-negative and from 28.7% to 13.2% in HIV-1-positive malaria patients (P < 0.001 for both). In patients with detectable but mostly asymptomatic parasitemia, CD4 count and, if HIV-1-infected, viral load at day 45 of follow-up were similar to those observed at enrollment. CONCLUSION: Interpretation of absolute CD4 count might be biased during or just after a clinical malaria episode. Therefore, in malaria-endemic areas, before taking any decision on the management of HIV-1-positive individuals, their malaria status should be assessed. PMID- 17079995 TI - Knowledge and use of HIV pre-exposure prophylaxis among attendees of minority gay pride events, 2004. PMID- 17079996 TI - The immunological response to highly active antiretroviral therapy is linked to CD4+ T-cell surface CCR5 density. PMID- 17079997 TI - Effects of growth hormone on visceral adipose tissue and dyslipidemia in HIV, an erratum. PMID- 17079998 TI - CD4 natural history and informative censoring in Sub-Saharan Africa. PMID- 17079999 TI - One-day quantitative cross-sectional study of family information time in 90 intensive care units in France. AB - RATIONALE: Providing family members with clear, honest, and timely information is a major task for intensive care unit physicians. Time spent informing families has been associated with effectiveness of information but has not been measured in specifically designed studies. OBJECTIVES: To measure time spent informing families of intensive care unit patients. METHODS: One-day cross-sectional study in 90 intensive care units in France. MEASUREMENTS: Clocked time spent by physicians informing the families of each of 951 patients hospitalized in the intensive care unit during a 24-hr period. MAIN RESULTS: Median family information time was 16 (interquartile range, 8-30) mins per patient, with 20% of the time spent explaining the diagnosis, 20% on explaining treatments, and 60% on explaining the prognosis. One third of the time was spent listening to family members. Multivariable analysis identified one factor associated with less information time (room with more than one bed) and seven factors associated with more information time, including five patient-related factors (surgery on the study day, higher Logistic Organ Dysfunction score, coma, mechanical ventilation, and worsening clinical status) and two family-related factors (first contact with family and interview with the spouse). Median information time was 20 (interquartile range, 10-39) mins when three factors were present and 106.5 (interquartile range, 103-110) mins when five were present. CONCLUSION: This study identifies factors associated with information time provided by critical care physicians to family members of critically ill patients. Whether information time correlates with communication difficulties or communication skills needs to be evaluated. Information time provided by residents and nurses should be studied. PMID- 17080000 TI - Mortality among patients admitted to intensive care units during weekday day shifts compared with "off" hours. AB - OBJECTIVE: To determine whether mortality rates among intensive care unit (ICU) patients differ according to the time of ICU admission, we compared the death rates for patients admitted during weekday day shifts and off hours (from 6:30 pm to 8:29 am the next day for night shifts, from Saturday 1:00 pm to Monday 8:29 am for weekends, and from 8:30 am to 8:29 am the following morning for public holidays). METHODS: Retrospective cohort study of data collected prospectively from 23 ICUs located in the Paris metropolitan region, France. Between January 2000 and December 2003, 51,643 patients were admitted to these ICUs. Patients were grouped according to their day and time of admission and compared using univariable and multivariable analyses. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Of the 51,643 patients admitted to ICUs, 33,857 (65.6%) were admitted during off hours. These latter patients were less critically ill than those admitted during day shifts, had fewer failed organs, required fewer support procedures, and their crude in-hospital mortality was lower (20.7 vs. 24.5%, p < .0001). After adjustment for initial disease severity, in-hospital mortality was not higher for off-hours admissions than weekday day admissions and even remained slightly lower (adjusted odds ratio, 0.93; 95% confidence interval, 0.87-0.98). CONCLUSIONS: Admission during off hours is common. In our ICUs, off-hours admissions were not associated with higher mortality and might even be associated with a lower death rate. PMID- 17080001 TI - Cardiac filling pressures are not appropriate to predict hemodynamic response to volume challenge. AB - OBJECTIVE: Values of central venous pressure of 8-12 mm Hg and of pulmonary artery occlusion pressure of 12-15 mm Hg have been proposed as volume resuscitation targets in recent international guidelines on management of severe sepsis. By analyzing a large number of volume challenges, our aim was to test the significance of the recommended target values in terms of prediction of volume responsiveness. DESIGN: Retrospective study. SETTING: A 24-bed medical intensive care unit. PATIENTS: All consecutive septic patients monitored with a pulmonary artery catheter who underwent a volume challenge between 2001 and 2004. INTERVENTION: None. MEASUREMENTS AND MAIN RESULTS: A total of 150 volume challenges in 96 patients were reviewed. In 65 instances, the volume challenge resulted in an increase in cardiac index of > or =15% (responders). The pre infusion central venous pressure was similar in responders and nonresponders (8 +/- 4 vs. 9 +/- 4 mm Hg). The pre-infusion pulmonary artery occlusion pressure was slightly lower in responders (10 +/- 4 vs. 11 +/- 4 mm Hg, p < .05). However, the significance of pulmonary artery occlusion pressure to predict fluid responsiveness was poor and similar to that of central venous pressure, as indicated by low values of areas under the receiver operating characteristic curves (0.58 and 0.63, respectively). A central venous pressure of <8 mm Hg and a pulmonary artery occlusion pressure of <12 mm Hg predicted volume responsiveness with a positive predictive value of only 47% and 54%, respectively. With the knowledge of a low stroke volume index (<30 mL.m), their positive predictive values were still unsatisfactory: 61% and 69%, respectively. When the combination of central venous pressure and pulmonary artery occlusion pressure was considered instead of either pressure alone, the degree of prediction of volume responsiveness was not improved. CONCLUSION: Our study demonstrates that cardiac filling pressures are poor predictors of fluid responsiveness in septic patients. Therefore, their use as targets for volume resuscitation must be discouraged, at least after the early phase of sepsis has concluded. PMID- 17080002 TI - Effect of acute kidney injury on weaning from mechanical ventilation in critically ill patients. AB - OBJECTIVES: Acute kidney injury (AKI) worsens outcome in various scenarios. We sought to investigate whether the occurrence of AKI has any effect on weaning from mechanical ventilation. DESIGN AND SETTING: Observational, retrospective study in a 23-bed medical/surgical intensive care unit (ICU) in a cancer hospital from January to December 2003. PATIENTS: The inclusion criterion was invasive mechanical ventilation for > or =48 hrs. AKI was defined as at least one measurement of serum creatinine of > or =1.5 mg/dL during the ICU stay. Patients were then separated into AKI and non-AKI patients (control group). The criterion for weaning was the combination of positive end-expiratory pressure of < or =8 cm H2O, pressure support of < or =10 cm H2O, and Fio2 of < or =0.4, with spontaneous breathing. The primary end point was duration of weaning and the secondary end points were rate of weaning failure, total length of mechanical ventilation, length of stay in the ICU, and ICU mortality. RESULTS: A total of 140 patients were studied: 93 with AKI and 47 controls. The groups were similar in regard to age, sex, and type of tumor. Diagnosis of acute lung injury/acute respiratory distress syndrome as cause of respiratory failure and Simplified Acute Physiology Score II at admission did not differ between groups. During ICU stay, AKI patients had markers of more severe disease: increased occurrence of severe sepsis or septic shock, higher number of antibiotics, and longer use of vasoactive drugs. The median (interquartile range) duration of mechanical ventilation (10 [6-17] vs. 7 [2-12] days, p = .017) and duration of weaning from mechanical ventilation (41 [16-97] vs. 21 [7-33.5] hrs, p = .018) were longer in AKI patients compared with control patients. Cox regression analysis demonstrated that a > or =85% increase in baseline serum creatinine (hazard rate, 2.30; 95% confidence interval, 1.30-4.08), oliguria (hazard rate, 2.51; 95% confidence interval, 1.24-5.08), and the number of antibiotics (hazard rate, 2.64; 95% confidence interval, 1.51-4.63) predicted longer duration of weaning. The length of ICU stay and ICU mortality rate were significantly greater in the AKI patients. After adjusting for Simplified Acute Physiology Score II, oliguria (odds ratio, 30.8; 95% confidence interval, 7.7-123.0) remained as a strong risk factor for mortality. CONCLUSION: This study shows that renal dysfunction has serious consequences in the duration of mechanical ventilation, weaning from mechanical ventilation, and mortality in critically ill cancer patients. PMID- 17080003 TI - Salmeterol enhances pulmonary fibrinolysis in healthy volunteers. AB - OBJECTIVE: Various lung diseases are associated with local activation of coagulation and concurrent inhibition of fibrinolysis. Although salmeterol, a beta2-adrenoceptor agonist with profound bronchodilatory properties, has been studied extensively, the effects of this compound on the pulmonary hemostatic balance are not elucidated. DESIGN: A single-blinded, placebo-controlled study. SETTING: University hospital and laboratory. SUBJECTS: A total of 32 human volunteers. INTERVENTIONS: Subjects inhaled 100 microg of salmeterol or placebo (t = -30 mins) followed by 100 microg of lipopolysaccharide (LPS) or normal saline (t = 0 mins; n = 8 per group). MEASUREMENTS AND MAIN RESULTS: Measurements were performed in bronchoalveolar lavage fluid obtained 6 hrs postchallenge. Inhalation of LPS enhanced pulmonary coagulation as determined by an increase in the concentrations of thrombin-antithrombin complexes, factor VIIa, and soluble tissue factor in bronchoalveolar lavage fluid (all p < .05 vs. saline). LPS concurrently inhibited pulmonary fibrinolysis, as reflected by a decrease in bronchoalveolar lavage fluid plasminogen activator activity together with an increase in plasminogen activator inhibitor type 1 (both p < .05 vs. saline). Moreover, LPS inhalation was associated with a suppression of the anticoagulant protein C pathway, as indicated by an increase in soluble thrombomodulin and decreases in protein C and activated protein C levels in bronchoalveolar lavage fluid (all p < .05 vs. saline). Salmeterol, either with or without LPS inhalation, enhanced fibrinolysis (plasminogen activator activity and tissue-type and urokinase-type plasminogen activator levels) but did not influence LPS induced changes in coagulation or the protein C pathway. CONCLUSIONS: Salmeterol has profibrinolytic properties in the normal lung and when applied in a model of sterile pulmonary inflammation. PMID- 17080004 TI - Early predictors for infection recurrence and death in patients with ventilator associated pneumonia. AB - OBJECTIVE: Early recognition of predictors of unfavorable evolution of ventilator associated pneumonia (VAP) might prompt therapeutic measures that might improve outcome. The objective of this study was to describe resolution of VAP variables and to determine early predictors of VAP recurrence and death. DESIGN AND SETTING: Description of the natural course of VAP resolution and multivariable analyses of predictors of VAP recurrence and death by day 28 after VAP onset based on the 401 patients included in the PNEUMA trial, a multiple-center, randomized study comparing 8 vs. 15 days of antibiotics for microbiologically proven VAP. Every patient included in that trial had received appropriate empirical antibiotics. MEASUREMENTS AND MAIN RESULTS: By day 28 after VAP onset, 27% of patients had VAP recurrence and 18% had died. On day 8 after VAP onset, predictors of VAP recurrence included intensive care unit admission Simplified Acute Physiology Score II (odds ratio [OR], 1.02), radiologic score (OR, 1.17), temperature (OR, 1.34), nonfermenting Gram-negative bacilli (OR, 2.00) or methicillin-resistant Staphylococcus aureus (OR, 2.50) as pathogens responsible for VAP, and mechanical ventilation dependency (OR, 2.08). Day 8 predictors of 28 day death were age (OR, 1.06), female sex (OR, 2.30), Sepsis-Related Organ Failure Assessment score (OR, 1.26), and nonfermenting Gram-negative bacilli (OR, 2.83) as pathogens responsible for VAP. However, the duration of antimicrobial therapy (8 vs. 15 days) was not associated with any of the studied adverse outcomes. CONCLUSIONS: For patients benefiting from appropriate empirical antibiotics for VAP, early predictors of infection recurrence or death included demographic characteristics, such as age or female sex, disease severity at VAP onset, nonfermenting Gram-negative bacilli or methicillin-resistant S. aureus as VAP-causative pathogens, prolonged mechanical ventilation dependency, persistent fever, and severity of lung injury. Future studies should attempt to determine whether specific diagnostic or therapeutic strategies could markedly improve VAP outcomes when early criteria for treatment failure are present. PMID- 17080005 TI - Renal arterial resistive index response to intraabdominal hypertension in a porcine model. AB - OBJECTIVE: The abdominal compartment syndrome is a potentially life-threatening condition with frequent renal involvement. There are few if any means of inferring subclinical effects before organ dysfunction. Because intrarenal pressure correlates with renal sonographic indices in other renal diseases, the purpose of this study was to determine the relationship between increasing intraabdominal hypertension and renal vascular flow velocities in a porcine model using renal Doppler ultrasound. DESIGN: Animal study. SETTING: University research laboratory. SUBJECTS: Eight anesthetized, mechanically ventilated, well hydrated, 30-kg female Yorkshire pigs. INTERVENTIONS: Intraabdominal hypertension was induced by instillation of warmed intraperitoneal saline through a midline laparoscopic port. Intraabdominal pressure (IAP) was continuously monitored directly from the peritoneum and indirectly from the bladder. IAP was varied from 0 to 50 mm Hg in increments of 5 mm Hg. At each IAP level, gray-scale, color, and spectral Doppler renal arcuate artery ultrasound was obtained and resistive index (RI) and peak airway pressure calculated. MEASUREMENTS AND MAIN RESULTS: Excellent agreement between direct and indirect IAP was found (bias, 0.032 mm Hg; 95% limits, -5.5 to 5.6 mm Hg). A linear relationship between RI and indirect IAP was observed and was defined by the regression equation: RI = 0.553 + 0.0104 x bladder pressure. There was a trend toward different RIs between left and right kidneys (p = .052) at the same IAP. RI varied in a linear fashion at low peak airway pressure and demonstrated an inflection point with steeper subsequent slope after peak airway pressure of 30 cm H2O. RI values rapidly returned to near baseline after abdominal decompression. CONCLUSIONS: In this model, the renal artery RI correlated strongly and linearly with the severity of intraabdominal hypertension, making renal Doppler ultrasound a potential noninvasive screening tool for the renal effects of intraabdominal hypertension. Further studies are warranted. PMID- 17080006 TI - Protein C -1641 AA is associated with decreased survival and more organ dysfunction in severe sepsis. AB - OBJECTIVE: Protein C contains an A/G polymorphism at position -1641 and a C/T polymorphism at -1654 associated with risk of deep venous thrombosis. We tested the hypothesis that these polymorphisms are associated with altered outcome in patients having severe sepsis, in which protein C is a central molecule. DESIGN: Prospective cohorts, gene-association study. SETTING: Tertiary care medical/surgical intensive care unit. PATIENTS: We first recruited a derivation cohort of patients having severe sepsis (n = 62). A second replication cohort was similarly defined but larger (n = 402). We tested for biological plausibility in a third cohort of post-cardiopulmonary bypass patients (n = 61). INTERVENTIONS: Patients were genotyped at protein C -1641 and -1654. MEASUREMENTS AND MAIN RESULTS: The primary outcome variable was survival in cohorts 1 and 2 and postoperative serum interleukin-6 concentration in cohort 3. Severity of individual organ dysfunctions and systemic inflammation were secondary outcome variables. In the first derivation cohort, the protein C -1641 AA genotype was associated with decreased 28-day survival (p < .05). This finding was confirmed in the much larger replication cohort of patients having severe sepsis (p = .028). In addition, the protein C -1641 AA genotype was associated with significantly more organ dysfunction and more clinical evidence of systemic inflammation (p < .05). Furthermore, the -1641 AA genotype was associated with increased serum interleukin-6 at 4 and 24 hrs after cardiopulmonary bypass (p = .024). There was no association of -1654 A/G with phenotype in any cohort. CONCLUSIONS: Protein C -1641 AA genotype is associated with decreased survival, more organ dysfunction, and more systemic inflammation in patients having severe sepsis and with increased interleukin-6 levels after cardiopulmonary bypass surgery. PMID- 17080007 TI - Correctable visual impairment among persons with diabetes--United States, 1999 2004. AB - Persons with diabetes are more likely to be visually impaired than persons without the disease. In 2005, CDC estimated that 14.6 million persons in the United States had diagnosed diabetes and an additional 6.2 million had undiagnosed diabetes. Despite the importance of detecting and treating vision problems caused by refractive errors (i.e., correctable visual impairment [CVI]), a limited number of studies have attempted to determine the proportion of persons with diabetes whose poor vision could be corrected with accurately prescribed glasses or contact lenses. To estimate that proportion, CDC analyzed 1999-2004 data from the National Health and Nutrition Examination Survey (NHANES). This report describes the results of that analysis, which indicated that among U.S. adults aged > or =20 years with diabetes, 11.0% had visual impairment (i.e., presenting visual acuity worse than 20/40 in their better-seeing eye while wearing glasses or contact lenses, if applicable) and approximately 65.5% of these cases of visual impairment were correctable. Health-care providers and persons with diabetes should be more aware that poor vision often is correctable and that visual corrections can reduce the risk for injury and improve the quality of life for persons with diabetes. PMID- 17080008 TI - Nutritional and health status of children during a food crisis--Niger, September 17-October 14, 2005. AB - Media attention in 2005 brought worldwide awareness to a food and nutrition crisis in the West Africa country of Niger (population 11.5 million in 2002). The United Nations World Food Programme estimated that 2.5 million persons living in farming and grazing areas in Niger were vulnerable to food insecurity (i.e., not having access at all times to enough food for an active, healthy lifestyle). Local surveys conducted in the Maradi and Tahoua administrative regions during April 2005 suggested critical levels (i.e., >15%) of global acute malnutrition (GAM) and greater mortality among Niger's estimated 2.7 million children aged <5 years than the emergency threshold (i.e., more than two deaths per 10,000 children per day). To help ensure a proportionate and timely response, the Government of Niger and the United Nations Children's Fund (UNICEF) collaborated with CDC to conduct an emergency survey that assessed the magnitude of malnutrition and recent illness among young children in Niger. This report summarizes the results of that survey, which determined that, among children aged 6-59 months, 15.3% had GAM; during the preceding 2 weeks, 72.0% had fever, and 49.1% had diarrhea. Among children aged 9-59 months, 33.7% had not been vaccinated for measles. Health officials in Niger took immediate action to improve availability of food, increase accessibility to medical treatment (for fever, diarrhea, and respiratory illness), and administer measles vaccinations along with vitamin A supplements to children who had not been vaccinated. PMID- 17080009 TI - The function of poliamine metabolism in prostate cancer. AB - In many developed countries prostate cancer is the second leading cause of cancer related death in human population. Prostate tissue is characterized by the highest level of polyamines among organs in human body, and it is even higher in prostate carcinomas. These ubiquitous molecules are synthesized by prostate epithelium and are involved in many biochemical processes including cell proliferation, cell cycle regulation and protein synthesis. In this review we made the attempt to discuss the functions of polyamines, their involvement in apoptosis and potential role as molecular biomarker for prostate cancer. Also we present recent data on generation of drugs, in particular, cyclin dependent kinase inhibitor, developed for therapy of prostate cancer. PMID- 17080010 TI - The role of cadherin/catenin complex in malignant melanoma. AB - In the present review article the role of cadherin/catenin complex in cases of malignant melanoma is discussed in some detail. Cadherins represent the most important superfamily of adhesion molecules with epithelial E-cadherin being the most studied. Its role in normal state as well as in cancer invasion and metastasis and some other pathologies is crucial. E-cadherin expression is altered in malignant melanomas and its downregulation or absence is associated with melanoma invasion and metastasis potential. A shift from E-cadherin expression to neural N-cadherin expression in melanocytes is also detected in malignant melanomas formation. In addition, a discussion regarding the role of placental P-cadherin and vascular endothelial VE-cadherin as well as the recently identified molecule of dysadherin, is attempted in brief. PMID- 17080011 TI - Molecular and clinical conditions associated with venous thromboembolism in oncological patients. AB - The association between cancer and thrombophilia has been known since 1865 since Trousseau described it. However in the last three decades an increased interest has been raised on this issue related to several molecular and condition that are involved in the daily management of oncological patients. This brief review has been focused on molecular conditions underlying cancer acquired thrombophilia then to further clinical aspects inducing thrombophilia in oncological patients such as surgery, chemotherapy, concomitant medical illness and inherited thrombophilia. PMID- 17080012 TI - Involvement of human beta-defensin-2 in intracellular signaling: in vitro study. AB - AIM: To analyze involvement of human beta-defensin-2 (hBD-2) in intracellular signaling in vitro. MATERIALS AND METHODS: A431cells were cultured in the presence of 1 microg/ml of recombinant hBD-2 and/or 10 ng/ml EGF. For evaluation of expression of mRNAs for p70S6 kinase, isoforms alpha and beta, RT-PCR analysis was applied. Expression and activity of p70S6K, phosphorylation of PDK1, ERK, JNK, p38 kinases and EGF receptor (EGFR) was evaluated using Western blot analysis. RESULTS: 30 min incubation of A431 cells with 1 mug/ml of hBD-2 didn't influence autophosphorylation level of EGFR, but resulted in activation of p70S6K, 12 h treatment - in prominently increased level of mRNA for alpha and beta-isoforms of p70S6 kinase, whilst 24 h treatment - in elevation of p70S6K synthesis on protein level. Up-stream kinase phosphorylating p70S6K, PDK1, is also phosporylated upon influence of exogenous hBD-2 in vitro. CONCLUSION: Our data point on the involvement of PDK1-p70S6K pathway in mediation of action of hBD-2 in A431 cells. PMID- 17080013 TI - Promotive effects of hyperthermia on the inhibition of DNA synthesis in ehrlich ascites tumor cells by eicosapentaenoic and docosahexaenoic acids. AB - AIM: To evaluate inhibitory effects of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on DNA synthesis in combination with hyperthermia in vitro. METHODS: A suspension of Ehrlich ascites tumor cells (EAT) was mixed with DHA or EPA in a glass tube, heated at 37 degrees C, 40 degrees C, or 42 degrees C for 1 h in a water bath, and cultured at 37 degrees C for 19 or 96 h. DNA synthesis was assayed by monitoring of the incorporation of [3H]-thymidine into the acid-insoluble fraction. DHA or EPA incorporated into EAT cells was extracted and measured by thin-layer chromatography and gas-liquid chromatography. RESULTS: The inhibition of DNA synthesis by EPA or DHA increased markedly upon the treatment at 42 degrees C and 40 degrees C compared to that at 37 degrees C. At 37 degrees C, inhibitory action of EPA was more potent than that of DHA at low concentrations (at 50 microM -- DNA synthesis level: EPA, 63.1%; DHA, 87.9%), whereas inhibitory action of DHA was higher at 150 muM (16.7%, 4.4%, ibid.). The effect of DHA compared to EPA was more marked at 40 degrees C (29.0%, 19.2% at 100 microM) or 42 degrees C (19.7%, 10.6% at 100 microM). Evaluation of DNA synthesis rate in the cells treated for 1 h by EPA or DHA with the next culturing of EAT cells for 19 h resulted in the enhanced inhibitory activity of EPA even at concentrations as low as 50 microM at either 37 degrees C (0.5%, 11.3%) or 42 degrees C (0.6%, 4.5%), which in these conditions was higher than that of DHA. At the same time the rate of incorporation of EPA in EAT cells at 37 degrees C or 42 degrees C was lower than that of DHA. CONCLUSION: Administration of DHA or EPA in vitro significantly inhibit DNA synthesis, and such effect is enhanced by combination of PUFAs with hyperthermia. PMID- 17080014 TI - Expression of clusterin, XIAP and survivin, and their changes by camptothecin (CPT) treatment in CPT-resistant PC-3 and CPT-sensitive LNCaP cells. AB - AIM: Clusterin and IAPs (inhibitor of apoptosis proteins), such as survivin and XIAP, are known to be related to chemo-resistance in several cancer cells. In the current study, we investigated their expression levels in human prostate cancer cell lines, LNCaP and PC-3 which are sensitive and resistant to camptothecin (CPT), topoisomerase I inhibitor, respectively. METHODS: LNCaP and PC-3 cells were cultured in the presence of CPT, cell death was evaluated using Hoechst 33342 and propidium iodide (PI) double staining. The expression of clusterin, XIAP and survivin on mRNA and protein levels was investigated by semi quantitative RT-PCR and Western blotting, respectively. RESULTS: Our data showed that 24 h treatment of LNCaP cells with 0.5 and 3.0 microM CPT resulted in higher number of apoptotic cells, than that in PC-3 cells. Western blot analysis revealed that the clusterin level in PC-3 cells was 5-fold higher than that in LNCaP cells. In contrast, XIAP expression level in PC-3 cells was lower than that in LNCaP cells, and survivin levels were similar in these two cell lines. Treatment with 0.5 and 3.0 microM CPT resulted in the reduced survivin and XIAP expression in both cell lines, while clusterin expression remained unchanged in LNCaP cells, but was increased in PC-3 cells. CONCLUSION: The results suggest that clusterin may take a greater part in CPT-resistance than survivin and XIAP in PC-3 cells. PMID- 17080015 TI - Differentiating effect of thalidomide and FGM-CS combination on HL-60 acute promyelocytic leukemia cells. AB - AIM: To investigate whether granulocyte-macrophage colony-stimulating factor (GM CSF) with or without thalidomide can induce apoptosis and differentiation of HL 60 acute promyelocytic leukemia cell line in vitro. METHODS: Effect of GM-CSF and thalidomide on proliferation of HL-60 cells was evaluated by MTT assay, cell cycle analysis was performed by propidium iodide staining approach and flow cytometry, and apoptosis rate was analyzed using FITC-conjugated annexin-V and FACScan flow cytometry. RESULTS: The study revealed that thalidomide alone at high concentrations inhibited HL-60 cell growth and induced apoptosis. Three days treatment of low-dose thalidomide in combination with GM-CSF induced marked terminal differentiation of HL-60 cells, as it was assessed by increased expression of differentiation antigens on cell surface. CONCLUSION: Treatment of HL-60 cells by low concentration of thalidomide combined with GM-CSF induced terminal differentiation of HL60 cells in vitro, which may be advantageous for the elaboration of novel therapeutic regimens in patients with differentiation inducible leukemias. PMID- 17080016 TI - The antitumor activity of thymoquinone and thymohydroquinone in vitro and in vivo. AB - AIM: The aim of the study was to investigate antitumor activity of thymoquinone (TQ) and thymohydroquinone (THQ) in vitro and in vivo. MATERIALS AND METHODS: In the in vitro experiments, L929 mouse broblasts and two tumor cell lines (squamous cell carcinoma (SCC VII) and fibrosarcoma (FsaR)) were used. The cells were cultured with 0.1 or 0.01 mg/ml TQ or THQ for 24 h, and cytotoxicity assay was performed with the use of crystal violet staining technique. For in vivo antitumor efficiency evaluation of new compounds two murine tumor models (fibrosarcoma (FsaR) and squamous cell carcinoma (SCC VII)) were used. The used dose was equal for both substances. Antitumor effect of 4 intratumoral injections of TQ and THQ at the dose of 5 mg/kg was evaluated by comparison of tumor growth kinetics between treated and control animals. RESULTS: In vitro study showed that TQ and THQ exhibit statistically significant cytotoxic activity (p less, similar 0.01). The cytotoxic activity was dose dependent and more expressed against tumor cells than against L929 fibroblasts. The result of antitumor activities of TQ and THQ in vivo reached TGI = 52% and it was statistically significant (p less, similar 0.05). CONCLUSION: The results indicate that THQ antitumor activity may be improved with further dose increase of the investigated substance. PMID- 17080017 TI - Immunohistochemical studies of protein kinase D (PKD) 2 expression in malignant human lymphomas. AB - AIM: To study the PKD2 expression, autophosphorylation and localization in reactive lymph nodes and tumors of lymphoid tissues. MATERIALS AND METHODS: Specific antibodies, which recognize PKD1/2 or PKD2 and autophosphorylated PKD1/2, were used for immunohistochemical and biochemical studies of tonsils, reactive lymph nodes, tumor samples of non-Hodgkin's lymphoma (NHL) and Hodgkin's lymphoma (HL). RESULTS: Immunohistochemical and biochemical analysis of PKD1 and PKD2 expression showed PKD2 expression in tonsils, reactive lymph nodes and tumor tissues from patients with NHL and HL. Furthermore, we were not able to reveal PKD1 expression in studied lymphoid tissues. In tonsils and reactive lymph nodes the PKD2 expression was detected in T and B cell zones with highest level in germinal centers of lymphoid follicles and the maximum level of autophosphorylation in the light zones of the germinal centers. We found that low level of PKD2 expression and autophosphorylation was characteristic feature for mantle cell lymphomas, Burkitt's lymphomas, and in 50% of CLL/small lymphocytic lymphomas. Lymphoma cells of germinal center origin and with activated B cell phenotype (diffuse large B cell lymphomas, HL) and anaplastic large cells lymphoma demonstrated the high level of PKD2 expression and autophosphorylation. CONCLUSIONS: The level of PKD2 expression and autophosphorylation in neoplastic cells corresponds to the expression pattern of this kinase in their normal analogs, and to the level of cell differentiation and activation. PMID- 17080018 TI - Antitumor action of lymphokin-activated cells of patients with soft tissue sarcomas and melanomas in dependence on expression of MHC classes I and II antigenes. AB - AIM: To study expression of major histocompatibility complex (MHC) classes and antigens and CD25, CD71, Ki-67, CD54, CD56, CD11b, PCNA on lymphocytes and tumor cells and antitumor action of lymphocytes activated with IL-2. MATERIALS AND METHODS: Tumor explants (soft tissue sarcoma, n = 20, melanoma, n = 25) were co cultivated in diffusion chambers with autologous lymphocytes; antitumor action was evaluated by morphologic patterns of explant's growth. Expression of CD25, CD71, Ki-67, CD54, CD56, CD11b, PCNA was evaluated by the method of indirect fluorescence using respective monoclonal antibodies. RESULTS: The highest antitumor action of lymphocytes toward soft tissue sarcoma and melanoma cells is observed if tumor cells are expressing MHC class I antigens. In the cases of soft tissue sarcoma no correlation between the level of antitumor activity of lymphocytes and expression of CD25, CD71, Ki-67, CD54, CD56, CD11b, PCNA has been found, whilst in the case of melanoma it is associated with the high level of CD11b expression. CONCLUSION: There is a direct correlation between sensitivity of soft tissue sarcoma and melanoma cells to action of lymphokin-activated killer cells and the level of MHC class I antigens. PMID- 17080019 TI - Relationship between HPV16/18 E6 and 53, 21WAF1, MDM2, Ki67 and cyclin D1 expression in esophageal squamous cell carcinoma: comparative study by using tissue microarray technology. AB - AIM: To investigate the role of human papillomavirus (HPV) HPV16/18 E6 oncogene in the carcinogenesis of esophageal cell carcinoma (ESCC). MATERIALS AND METHODS: Tissue microarray (TMA) block was constructed from 60 cases of paraffin-embedded ESCC tissues and pair-matched controls (adjacent normal epithelium). Immunohistochemistry (IHC) methods were applied to detect the expression of HPV16/18 E6, p53, p21(WAF1), MDM2, Ki67 and cyclin D1 proteins on TMA slides. In situ hybridization (ISH) targeting HPV gene was also used. RESULTS: In ESCC samples, 18.3% (11/60) were revealed HPV16/18 E6 positive by IHC, while 40.0% (24/60) HPV positive by ISH; HPV16/18 E6 expression was significantly higher than that of control samples. In ESCC samples, the expressions of p53, p21(WAF1), Cyclin D1, MDM2 and Ki67 were recorded in 60.0% (36/60), 40.0% (24/60), 51.7% (31/60), 65.0% (39/60) and 88.3% (53/60) cases respectively, In ESCC samples, p53, MDM2 and Ki67 expression correlated with the HPV16/18 E6 expression (p less, similar 0.01), p21(WAF1) expression - with these of MDM2 and cyclin D1 (p less, similar 0.01) whilst expression of Ki67 - with ESCC grade (p less, similar 0.01). CONCLUSION: HPV might be one of etiological factor of esophageal carcinoma in Shantou, China. p53, MDM2 proteins may play important roles in the pathogenesis of HPV-associated ESCC. PMID- 17080020 TI - Leptin and resistin levels in serum of patients with hematologic malignancies: correlation with clinical characteristics. AB - AIM: To evaluate leptin and resistin levels in patients with various hematologic malignancies. METHODS: We included 21 patients with lymphoma, 14 with multiple myeloma (MM), 14 with acute leukemia, 13 with chronic lymphocytic leukemia (CLL), and 25 healthy control subjects into our study. The subjects' body mass indexes (BMI) were calculated; hematological and acute phase response parameters, serum lipid were determined; serum leptin and resistin levels were determined by ELISA. RESULTS: Serum leptin level was significantly increased in CLL and MM groups when compared to the control group (p less, similar 0.01). Resistin level was significantly higher in lymphoma patients than in CLL, acute leukemia and control groups (p less, similar 0.01). In the control group, leptin level was negatively correlated with hemoglobin level (r = -0.44, p = 0.047); and in all patients with hematologic malignancies, leptin level was correlated with BMI (r = 0.32, p = 0.02). Leptin in lymphoma subjects correlated with hemoglobin level (r = 0.64, p = 0.005), resistin level correlated with the platelet count in patients with hematologic malignancies (r = 0.26, p = 0.044). In addition, leptin level had negative correlations with international prognostic score (IPS) in Hodgkin lymphoma (r = -0.9, p = 0.002) and with international prognostic index (IPI) in non-Hodgkin lymphoma (r = -0.77, p = 0.03). In CLL patients, leptin level had a correlation with the poor prognostic marker - CD38 level (r = 0.68, p = 0.03). CONCLUSION: We found higher leptin levels in MM and CLL patients, and higher resistin levels in lymphoma patients: this fact demonstrates that changes in adipose tissue and metabolism occur in these disease states. PMID- 17080021 TI - Upper extremity DVT in oncological patients: analysis of risk factors. Data from the RIETE registry. AB - AIM: The aim of the study is to up date informations on the clinical characteristics and outcome of patients with upper-extremity deep vein thrombosis (DVT) from the Informatised Registry on Venous Thromboembolism (RIETE). METHODS: RIETE is an ongoing registry of consecutive patients with symptomatic, objectively confirmed, acute venous thromboembolism. In this analysis the clinical characteristics and 3-month outcome of all cancer patients with upper extremity DVT were evaluated. RESULTS: Up to February 2006, a total of 14,391 patients with symptomatic, objectively confirmed acute venous thromboembolism had been enrolled in RIETE. Of the 2,945 patients with active cancer 196 (6.7%) had arm DVT: 104 had catheter-associated DVT. Most cancer patients with arm DVT were males, younger than 65, and had a low incidence of additional risk factors or underlying diseases. Twenty of them (10%) had symptomatic pulmonary embolism (PE). Most patients were treated with low-molecular-weigh heparin, both initially (94%) and after discharge (75%). During the 3-month follow-up period 12 patients (6.1%) developed VTE recurrences (PE 6, DVT 6), 8 (4.1%) had major bleeding (fatal in 3), 43 (22%) died. CONCLUSIONS: Our data from the RIETE registry show that upper limb DVT is a serious complication in patients with cancer, with a high incidence of recurrences and bleeding complications. PMID- 17080022 TI - Crossing chromosomes in pleomorphic sarcoma. PMID- 17080023 TI - Granulocyte colony-stimulating factor (G-CSF): a novel anticancer therapy based on the "universal dynamics of tumor growth"? AB - It has recently been proposed that all solid tumors exhibit the same growth dynamics. This hypothesis, developed by Bru and coworkers and called the universal dynamics of tumor growth, says that the main mechanism responsible for tumor progression is cell diffusion on the tumor border. The authors of this hypothesis claim that, by inducing strong neutrophilia around the tumor, this dynamic can be changed; neutrophils would locate themselves to eliminate cell diffusion on the tumor border therefore inhibiting tumor growth. The authors suggest that this approach may be exploited to develop effective anticancer strategies, and they have recently reported the possible cure of a 56-year-old patient with advanced hepatocarcinoma treated with granulocyte colony-stimulating factor (G-CSF), a key regulator of neutrophil production. The present report shows evidence that suggests that it is very unlikely that neutrophil-induced cancer cell death is mediated by a mechanical impediment at the tumor border. Furthermore, it is shown that the induction of neutrophilia is not a new anticancer strategy based on the "universal dynamics of tumor growth", but a known approach that has been widely explored along the years. The merits of G-CSF for being tested in clinical trials with cancer patients are finally evaluated. PMID- 17080024 TI - Putting the "landscape" in landscape genetics. AB - Landscape genetics has emerged as a new research area that integrates population genetics, landscape ecology and spatial statistics. Researchers in this field can combine the high resolution of genetic markers with spatial data and a variety of statistical methods to evaluate the role that landscape variables play in shaping genetic diversity and population structure. While interest in this research area is growing rapidly, our ability to fully utilize landscape data, test explicit hypotheses and truly integrate these diverse disciplines has lagged behind. Part of the current challenge in the development of the field of landscape genetics is bridging the communication and knowledge gap between these highly specific and technical disciplines. The goal of this review is to help bridge this gap by exposing geneticists to terminology, sampling methods and analysis techniques widely used in landscape ecology and spatial statistics but rarely addressed in the genetics literature. We offer a definition for the term "landscape genetics", provide an overview of the landscape genetics literature, give guidelines for appropriate sampling design and useful analysis techniques, and discuss future directions in the field. We hope, this review will stimulate increased dialog and enhance interdisciplinary collaborations advancing this exciting new field. PMID- 17080025 TI - Drosophila melanogaster as a model for studying protein-encoding genes that are resident in constitutive heterochromatin. AB - The organization of chromosomes into euchromatin and heterochromatin is one of the most enigmatic aspects of genome evolution. For a long time, heterochromatin was considered to be a genomic wasteland, incompatible with gene expression. However, recent studies--primarily conducted in Drosophila melanogaster--have shown that this peculiar genomic component performs important cellular functions and carries essential genes. New research on the molecular organization, function and evolution of heterochromatin has been facilitated by the sequencing and annotation of heterochromatic DNA. About 450 predicted genes have been identified in the heterochromatin of D. melanogaster, indicating that the number of active genes is higher than had been suggested by genetic analysis. Most of the essential genes are still unknown at the molecular level, and a detailed functional analysis of the predicted genes is difficult owing to the lack of mutant alleles. Far from being a peculiarity of Drosophila, heterochromatic genes have also been found in Saccharomyces cerevisiae, Schizosaccharomyces pombe, Oryza sativa and Arabidopsis thaliana, as well as in humans. The presence of expressed genes in heterochromatin seems paradoxical because they appear to function in an environment that has been considered incompatible with gene expression. In the future, genetic, functional genomic and proteomic analyses will offer powerful approaches with which to explore the functions of heterochromatic genes and to elucidate the mechanisms driving their expression. PMID- 17080026 TI - A cost-effectiveness approach to the qualification and acceptance of biomarkers. AB - The flow of new medicines to patients depends on the development of new biomarkers and their correct interpretation, yet there are no widely accepted and practically applicable criteria that facilitate adequate biomarker qualification. As a result, case-by-case qualifications are based on subjective assessments that do not lead to optimal decisions for patients, which have contributed to the 'stagnation' in drug productivity identified by the FDA. An alternative is to qualify biomarkers in terms of cost effectiveness using a set of principles that enable the evaluation of biomarkers even with incomplete knowledge. This approach could minimize harm to patients, improve access to medicines and reduce healthcare costs. PMID- 17080027 TI - PI3Kgamma inhibition: towards an 'aspirin of the 21st century'? AB - Class IB phosphatidylinositol 3-kinase p110gamma (PI3Kgamma) has gained increasing attention as a promising drug target for the treatment of inflammatory disease. Extensive target-validation data are available, which are derived from studies using both pharmacological and genetic tools. More recent findings have uncovered further therapeutic applications for PI3Kgamma inhibitors, opening up potentially huge opportunities for these drugs. Several companies have been pursuing small-molecule PI3Kgamma inhibitor projects, but none of them has progressed to the clinic yet. Here, we discuss the insights gained so far and the main challenges that are emerging on the path to developing PI3Kgamma inhibitors for the treatment of human disease. PMID- 17080028 TI - The obesity pipeline: current strategies in the development of anti-obesity drugs. AB - This review provides a summary of currently available pharmaceutical therapies for the treatment of obesity, along with an overview of the pipeline of products currently in development, and the key mechanisms on which the major development candidates are based. In particular, the recent increase in understanding of the role of gut peptides in energy homeostasis is highlighted as a promising source of potential future obesity therapies. PMID- 17080029 TI - Issues in the design and implementation of vaccine trials in less developed countries. AB - There is a growing need for vaccine trials in developing countries. This need arises from the fact that some vaccines do not perform as well in developing as in industrialized country populations and because some newly developed candidates target diseases found only in less-developed countries. Here we discuss several key issues in the design and implementation of vaccine trials in less-developed countries. These include the phasing of vaccine trials on safety, immunogenicity and efficacy to achieve licensure; the recent use of trials to obtain other information crucial to the ultimate deployment of vaccines (such as immunological correlates of protection, indirect vaccine effects, and practical information on the feasibility, costs and acceptability of vaccine introduction); and several ethical issues that have arisen in connection with trials done in developing countries. PMID- 17080030 TI - Innovative lead discovery strategies for tropical diseases. AB - Lead discovery is currently a key bottleneck in the pipeline for much-needed novel drugs for tropical diseases such as malaria, tuberculosis, African sleeping sickness, leishmaniasis and Chagas disease. Here, we discuss the different approaches to lead discovery for tropical diseases and emphasize a coordination strategy that involves highly integrated partnerships and networks between scientists in academic institutions and industry in both wealthy industrialized countries and disease-endemic countries. This strategy offers the promise of reducing the inherently high attrition rate of the early stages of discovery research, thereby increasing the chances of success and enhancing cost effectiveness. PMID- 17080031 TI - Drug discovery in dementia: the role of rodent models. AB - Recent advances in the understanding of the pathophysiological mechanisms underlying Alzheimer's disease have pointed to novel strategies for drug development. Animal models have contributed considerably to these advances, and will have a key role in the evaluation of therapeutics that could have the potential not just to alleviate the dementia associated with Alzheimer's disease, but to modify the disease process. Here, we summarize and critically evaluate current rodent models of dementia, and discuss their role in drug discovery and development. PMID- 17080032 TI - Geochemistry: how well can Pb isotopes date core formation? AB - Timescale and the physics of planetary core formation are essential constraints for models of Earth's accretion and early differentiation. Wood and Halliday use the apparent mismatch in core-formation dates determined from tungsten (W) and lead (Pb) chrono-meters to argue for a two-stage core formation, involving an early phase of metal segregation followed by a protracted episode of sulphide melt addition. However, we show here that crust-;mantle Pb isotope systematics do not require diachronous core formation. Our observations indicate that very early (< or = 35 Myr) core formation and planet accretion remain the most plausible scenario. PMID- 17080033 TI - Geochemistry: does U-Pb date Earth's core formation? AB - Constraining the timing of the formation of Earth's core, which defines the birth of our planet, is essential for understanding the early evolution of Earth-like planets. Wood and Halliday and Halliday discuss the apparent discrepancy between the U-Pb (60-80 Myr) and Hf-W clocks (30 Myr) in determining the timescale of Earth's accretion and core formation. We find that the information the authors present is at times contradictory (for example, compare Fig. 1 in ref. 1 with Fig. 1 in ref. 2) and confusing and could suggest that the U-Pb clock constrains core formation better than the Hf-W system. Here we point out the limitations of the U-Pb system and show that the U-Pb age cannot be used to argue for protracted accretion and/or core formation (>50 Myr) because this clock only records the processes that occurred during the last 1% of Earth's accretion and core formation in the Wood and Halliday mechanism. PMID- 17080038 TI - Science and the Islamists. PMID- 17080039 TI - Enough biodefence. PMID- 17080040 TI - A global call to arms. PMID- 17080043 TI - How much will it cost to save the world? PMID- 17080046 TI - Search for alien signals stalls for want of cash. PMID- 17080044 TI - Ethiopian plan for Lucy tour splits museums. PMID- 17080047 TI - A votre sante: now in pill form? PMID- 17080048 TI - Safer embryo tests could boost IVF pregnancy rates. PMID- 17080049 TI - Hwang takes the stand at fraud trial. PMID- 17080050 TI - NIH researchers fear ethics rules will hit recruitment. PMID- 17080052 TI - Drilling for nanotech gold. PMID- 17080055 TI - Islam and science: ambition and neglect. PMID- 17080056 TI - Islam and Science: the Islamic world. PMID- 17080057 TI - Islam and Science: an Islamist revolution. PMID- 17080058 TI - Islam and Science: the data gap. PMID- 17080059 TI - Islam and Science: oil rich, science poor. PMID- 17080060 TI - Islam and Science: Q and A The reformer. Mostafa Moin interviewed by Declan Butler. PMID- 17080063 TI - No room for complacency on drug resistance in Africa. PMID- 17080064 TI - The daunting process of MIAME. PMID- 17080065 TI - Dishing a modern myth about microbes. PMID- 17080066 TI - Islam and Science: steps towards reform. PMID- 17080067 TI - Islam and science: where are the new patrons of science? PMID- 17080073 TI - Photonics: a cooling light breeze. PMID- 17080075 TI - Geophysics: same old magnetism. PMID- 17080074 TI - Evolutionary biology: to work or not to work. PMID- 17080077 TI - Cancer biology: second step to retinal tumours. PMID- 17080078 TI - Physical chemistry: porous solids get organized. PMID- 17080079 TI - Neurobiology: crossed circuits. PMID- 17080080 TI - Materials science: qubits in the pink. PMID- 17080081 TI - Enforced altruism in insect societies. AB - Cooperation among workers and their seeming altruism result from strict policing by nestmates. PMID- 17080082 TI - Proterozoic low orbital obliquity and axial-dipolar geomagnetic field from evaporite palaeolatitudes. AB - Palaeomagnetism of climatically sensitive sedimentary rock types, such as glacial deposits and evaporites, can test the uniformitarianism of ancient geomagnetic fields and palaeoclimate zones. Proterozoic glacial deposits laid down in near equatorial palaeomagnetic latitudes can be explained by 'snowball Earth' episodes, high orbital obliquity or markedly non-uniformitarian geomagnetic fields. Here I present a global palaeomagnetic compilation of the Earth's entire basin-scale evaporite record. Magnetic inclinations are consistent with low orbital obliquity and a geocentric-axial-dipole magnetic field for most of the past two billion years, and the snowball Earth hypothesis accordingly remains the most viable model for low-latitude Proterozoic ice ages. Efforts to reconstruct Proterozoic supercontinents are strengthened by this demonstration of a consistently axial and dipolar geomagnetic reference frame, which itself implies stability of geodynamo processes on billion-year timescales. PMID- 17080083 TI - Inactivation of the p53 pathway in retinoblastoma. AB - Most human tumours have genetic mutations in their Rb and p53 pathways, but retinoblastoma is thought to be an exception. Studies suggest that retinoblastomas, which initiate with mutations in the gene retinoblastoma 1 (RB1), bypass the p53 pathway because they arise from intrinsically death resistant cells during retinal development. In contrast to this prevailing theory, here we show that the tumour surveillance pathway mediated by Arf, MDM2, MDMX and p53 is activated after loss of RB1 during retinogenesis. RB1-deficient retinoblasts undergo p53-mediated apoptosis and exit the cell cycle. Subsequently, amplification of the MDMX gene and increased expression of MDMX protein are strongly selected for during tumour progression as a mechanism to suppress the p53 response in RB1-deficient retinal cells. Our data provide evidence that the p53 pathway is inactivated in retinoblastoma and that this cancer does not originate from intrinsically death-resistant cells as previously thought. In addition, they support the idea that MDMX is a specific chemotherapeutic target for treating retinoblastoma. PMID- 17080084 TI - Self-cooling of a micromirror by radiation pressure. AB - Cooling of mechanical resonators is currently a popular topic in many fields of physics including ultra-high precision measurements, detection of gravitational waves and the study of the transition between classical and quantum behaviour of a mechanical system. Here we report the observation of self-cooling of a micromirror by radiation pressure inside a high-finesse optical cavity. In essence, changes in intensity in a detuned cavity, as caused by the thermal vibration of the mirror, provide the mechanism for entropy flow from the mirror's oscillatory motion to the low-entropy cavity field. The crucial coupling between radiation and mechanical motion was made possible by producing free-standing micromirrors of low mass (m approximately 400 ng), high reflectance (more than 99.6%) and high mechanical quality (Q approximately 10,000). We observe cooling of the mechanical oscillator by a factor of more than 30; that is, from room temperature to below 10 K. In addition to purely photothermal effects we identify radiation pressure as a relevant mechanism responsible for the cooling. In contrast with earlier experiments, our technique does not need any active feedback. We expect that improvements of our method will permit cooling ratios beyond 1,000 and will thus possibly enable cooling all the way down to the quantum mechanical ground state of the micromirror. PMID- 17080085 TI - Radiation-pressure cooling and optomechanical instability of a micromirror. AB - Recent table-top optical interferometry experiments and advances in gravitational wave detectors have demonstrated the capability of optical interferometry to detect displacements with high sensitivity. Operation at higher powers will be crucial for further sensitivity enhancement, but dynamical effects caused by radiation pressure on the interferometer mirrors must be taken into account, and the appearance of optomechanical instabilities may jeopardize the stable operation of the next generation of interferometers. These instabilities are the result of a nonlinear coupling between the motion of the mirrors and the optical field, which modifies the effective dynamics of the mirror. Such 'optical spring' effects have already been demonstrated for the mechanical damping of an electromagnetic waveguide with a moving wall, the resonance frequency of a specially designed flexure oscillator, and the optomechanical instability of a silica microtoroidal resonator. Here we present an experiment where a micromechanical resonator is used as a mirror in a very high-finesse optical cavity, and its displacements are monitored with unprecedented sensitivity. By detuning the laser frequency with respect to the cavity resonance, we have observed a drastic cooling of the microresonator by intracavity radiation pressure, down to an effective temperature of 10 kelvin. For opposite detuning, efficient heating is observed, as well as a radiation-pressure-induced instability of the resonator. Further experimental progress and cryogenic operation may lead to the experimental observation of the quantum ground state of a micromechanical resonator, either by passive or active cooling techniques. PMID- 17080086 TI - Sub-kelvin optical cooling of a micromechanical resonator. AB - Micromechanical resonators, when cooled down to near their ground state, can be used to explore quantum effects such as superposition and entanglement at a macroscopic scale. Previously, it has been proposed to use electronic feedback to cool a high frequency (10 MHz) resonator to near its ground state. In other work, a low frequency resonator was cooled from room temperature to 18 K by passive optical feedback. Additionally, active optical feedback of atomic force microscope cantilevers has been used to modify their response characteristics, and cooling to approximately 2 K has been measured. Here we demonstrate active optical feedback cooling to 135 +/- 15 mK of a micromechanical resonator integrated with a high-quality optical resonator. Additionally, we show that the scheme should be applicable at cryogenic base temperatures, allowing cooling to near the ground state that is required for quantum experiments--near 100 nK for a kHz oscillator. PMID- 17080087 TI - Complex zeolite structure solved by combining powder diffraction and electron microscopy. AB - Many industrially important materials, ranging from ceramics to catalysts to pharmaceuticals, are polycrystalline and cannot be grown as single crystals. This means that non-conventional methods of structure analysis must be applied to obtain the structural information that is fundamental to the understanding of the properties of these materials. Electron microscopy might appear to be a natural approach, but only relatively simple structures have been solved by this route. Powder diffraction is another obvious option, but the overlap of reflections with similar diffraction angles causes an ambiguity in the relative intensities of those reflections. Various ways of overcoming or circumventing this problem have been developed, and several of these involve incorporating chemical information into the structure determination process. For complex zeolite structures, the FOCUS algorithm has proved to be effective. Because it operates in both real and reciprocal space, phase information obtained from high-resolution transmission electron microscopy images can be incorporated directly into this algorithm in a simple way. Here we show that by doing so, the complexity limit can be extended much further. The power of this approach has been demonstrated with the solution of the structure of the zeolite TNU-9 (|H9.3|[Al9.3Si182.7O384]; ref. 10) with 24 topologically distinct (Si,Al) atoms and 52 such O atoms. For comparison, ITQ-22 (ref. 11), the most complex zeolite known to date, has 16 topologically distinct (Si,Ge) atoms. PMID- 17080088 TI - 10Be evidence for the Matuyama-Brunhes geomagnetic reversal in the EPICA Dome C ice core. AB - An ice core drilled at Dome C, Antarctica, is the oldest ice core so far retrieved. On the basis of ice flow modelling and a comparison between the deuterium signal in the ice with climate records from marine sediment cores, the ice at a depth of 3,190 m in the Dome C core is believed to have been deposited around 800,000 years ago, offering a rare opportunity to study climatic and environmental conditions over this time period. However, an independent determination of this age is important because the deuterium profile below a depth of 3,190 m depth does not show the expected correlation with the marine record. Here we present evidence for enhanced 10Be deposition in the ice at 3,160 3,170 m, which we interpret as a result of the low dipole field strength during the Matuyama-Brunhes geomagnetic reversal, which occurred about 780,000 years ago. If correct, this provides a crucial tie point between ice cores, marine cores and a radiometric timescale. PMID- 17080089 TI - Post-mating sexual selection increases lifetime fitness of polyandrous females in the wild. AB - Females often mate with several males before producing offspring. Field studies of vertebrates suggest, and laboratory experiments on invertebrates confirm, that even when males provide no material benefits, polyandry can enhance offspring survival. This enhancement is widely attributed to genetic benefits that arise whenever paternity is biased towards males that sire more viable offspring. Field studies suggest that post-mating sexual selection biases fertilization towards genetically more compatible males and one controlled experiment has shown that, when females mate with close kin, polyandry reduces the relative number of inbred offspring. Another potential genetic benefit of polyandry is that it increases offspring survival because males with more competitive ejaculates sire more viable offspring. Surprisingly, however, there is no unequivocal evidence for this process. Here, by experimentally assigning mates to females, we show that polyandry greatly increases offspring survival in the Australian marsupial Antechinus stuartii. DNA profiling shows that males that gain high paternity under sperm competition sire offspring that are more viable. This beneficial effect occurs in both the laboratory and the wild. Crucially, there are no confounding non-genetic maternal effects that could arise if polyandry increases female investment in a particular reproductive event because A. stuartii is effectively semelparous. Our results therefore show that polyandry improves female lifetime fitness in nature. The threefold increase in offspring survival is not negated by a decline in maternal lifespan and is too large to be offset by an equivalent decline in the reproductive performance of surviving offspring. PMID- 17080090 TI - Global distribution and conservation of rare and threatened vertebrates. AB - Global conservation strategies commonly assume that different taxonomic groups show congruent geographical patterns of diversity, and that the distribution of extinction-prone species in one group can therefore act as a surrogate for vulnerable species in other groups when conservation decisions are being made. The validity of these assumptions remains unclear, however, because previous tests have been limited in both geographical and taxonomic extent. Here we use a database on the global distribution of 19,349 living bird, mammal and amphibian species to show that, although the distribution of overall species richness is very similar among these groups, congruence in the distribution of rare and threatened species is markedly lower. Congruence is especially low among the very rarest species. Cross-taxon congruence is also highly scale dependent, being particularly low at the finer spatial resolutions relevant to real protected areas. 'Hotspots' of rarity and threat are therefore largely non-overlapping across groups, as are areas chosen to maximize species complementarity. Overall, our results indicate that 'silver-bullet' conservation strategies alone will not deliver efficient conservation solutions. Instead, priority areas for biodiversity conservation must be based on high-resolution data from multiple taxa. PMID- 17080092 TI - XIAP deficiency in humans causes an X-linked lymphoproliferative syndrome. AB - The homeostasis of the immune response requires tight regulation of the proliferation and apoptosis of activated lymphocytes. In humans, defects in immune homeostasis result in lymphoproliferation disorders including autoimmunity, haemophagocytic lymphohystiocytosis and lymphomas. The X-linked lymphoproliferative syndrome (XLP) is a rare, inherited immunodeficiency that is characterized by lymphohystiocytosis, hypogammaglobulinaemia and lymphomas, and that usually develops in response to infection with Epstein-Barr virus (EBV). Mutations in the signalling lymphocyte activation molecule (SLAM)-associated protein SAP, a signalling adaptor molecule, underlie 60% of cases of familial XLP. Here, we identify mutations in the gene that encodes the X-linked inhibitor of-apoptosis XIAP (also termed BIRC4) in patients with XLP from three families without mutations in SAP. These mutations lead to defective expression of XIAP. We show that apoptosis of lymphocytes from XIAP-deficient patients is enhanced in response to various stimuli including the T-cell antigen receptor (TCR)-CD3 complex, the death receptor CD95 (also termed Fas or Apo-1) and the TNF associated apoptosis-inducing ligand receptor (TRAIL-R). We also found that XIAP deficient patients, like SAP-deficient patients, have low numbers of natural killer T-lymphocytes (NKT cells), indicating that XIAP is required for the survival and/or differentiation of NKT cells. The observation that XIAP deficiency and SAP-deficiency are both associated with a defect in NKT cells strengthens the hypothesis that NKT cells have a key role in the immune response to EBV. Furthermore, by identifying an XLP immunodeficiency that is caused by mutations in XIAP, we show that XIAP is a potent regulator of lymphocyte homeostasis in vivo. PMID- 17080091 TI - Insights from the genome of the biotrophic fungal plant pathogen Ustilago maydis. AB - Ustilago maydis is a ubiquitous pathogen of maize and a well-established model organism for the study of plant-microbe interactions. This basidiomycete fungus does not use aggressive virulence strategies to kill its host. U. maydis belongs to the group of biotrophic parasites (the smuts) that depend on living tissue for proliferation and development. Here we report the genome sequence for a member of this economically important group of biotrophic fungi. The 20.5-million-base U. maydis genome assembly contains 6,902 predicted protein-encoding genes and lacks pathogenicity signatures found in the genomes of aggressive pathogenic fungi, for example a battery of cell-wall-degrading enzymes. However, we detected unexpected genomic features responsible for the pathogenicity of this organism. Specifically, we found 12 clusters of genes encoding small secreted proteins with unknown function. A significant fraction of these genes exists in small gene families. Expression analysis showed that most of the genes contained in these clusters are regulated together and induced in infected tissue. Deletion of individual clusters altered the virulence of U. maydis in five cases, ranging from a complete lack of symptoms to hypervirulence. Despite years of research into the mechanism of pathogenicity in U. maydis, no 'true' virulence factors had been previously identified. Thus, the discovery of the secreted protein gene clusters and the functional demonstration of their decisive role in the infection process illuminate previously unknown mechanisms of pathogenicity operating in biotrophic fungi. Genomic analysis is, similarly, likely to open up new avenues for the discovery of virulence determinants in other pathogens. PMID- 17080093 TI - Erectile dysfunction influences the subsequent incidence of lower urinary tract symptoms and bother. AB - It is unclear whether lower urinary tract symptoms (LUTS) cause erectile dysfunction (ED) independently or through common underlying pathophysiology and shared risk factors. The aim of this study was to investigate the effect of ED on the incidence of frequency and bother of LUTS. Target population consisted of men aged 50, 60 or 70 years residing in the study area in Finland in 1994. Questionnaires were mailed to 3143 men in 1994 and to 2837 of them 5 years later. The follow-up sample comprised 1683 men who responded to both baseline and follow up surveys. ED was assessed by two questions on subject's ability to achieve or maintain an erection sufficient for intercourse and LUTS by the Danish Prostatic Symptom Score questionnaire. A dose-response relation was found between the severity of ED at baseline and the incidence of LUTS or bother during follow-up. After adjustment for the confounders, the incidence rate ratio (RR) of LUTS was higher in men with moderate (RR 1.5, 95% confidence interval (CI) 1.0-2.3) or severe ED (RR 2.3, 95% CI 1.4-3.8) than in those free of ED at entry. Compared with men free of ED at baseline, the RRs of urinary bother were 1.6 (95% CI 1.1 2.4), 1.9 (95% CI 1.1-3.2) and 2.2 (95% CI 1.1-4.3) for minimal, moderate or severe ED, respectively. In summary, ED is associated with an increased incidence of LUTS and bother. ED and LUTS may have a common underlying pathophysiology or shared risk factors. PMID- 17080094 TI - Neonatal Candida meningitis: significance of cerebrospinal fluid parameters and blood cultures. AB - OBJECTIVE: The purpose of this study was to examine the frequency of normal cerebrospinal fluid (CSF) parameters in Candida meningitis and the proportion of candidemia associated with Candida meningitis. STUDY DESIGN: We evaluated the initial lumbar puncture results from infants discharged from 150 Neonatal Intensive Care Units between 1997 and 2004. Candida meningitis was diagnosed by a positive CSF culture or positive Gram stain for yeast. We calculated two-tailed P values using non-parametric testing, Mann-Whitney, Kruskal-Wallis or Fisher's exact tests where appropriate. RESULTS: Twenty infants had culture-positive Candida meningitis. Normal CSF parameters were found in 43% (3/7) of the infants with Candida meningitis and only 37% (7/19) of them had positive blood cultures for Candida. CONCLUSION: Normal CSF parameters do not exclude the diagnosis of neonatal Candida meningitis. The majority of infants in this cohort with Candida meningitis did not have evidence of candidemia at the time of diagnosis. PMID- 17080095 TI - Second trimester placental location as a predictor of an adverse pregnancy outcome. AB - OBJECTIVE: To determine if the second trimester placental location is associated with perinatal outcomes. MATERIALS AND METHODS: Observational study of placental location and the subsequent risk of an adverse pregnancy outcome. Placental location was divided into three categories, low, high lateral and high fundal. RESULTS: There were 3336 pregnancies analyzed in this study. Low implantation sites had a greater risk of preterm labor (odds ratio (OR) 1.70, 95% confidence interval (CI) 1.38 to 2.90, P<0.001), preterm delivery (OR 1.86, 95% CI 1.36 to 2.54, P<0.001), fewer fetuses with macrosomia (OR 0.56, 95% CI 0.38 to 0.83, P=0.010) and reduced risk of postpartum hemorrhage (OR 0.56, 95% CI 0.46 to 0.95, P=0.026). High lateral implantations had a greater risk of low 1-min (OR 1.80, 95% CI 1.11 to 2.93, P=0.017) and 5-min (OR 3.49, 95% CI 1.46 to 8.36, P=0.005) Apgar scores. CONCLUSIONS: Low placental implantation was associated with an increased risk of preterm labor, preterm delivery and a reduced risk of postpartum hemorrhage, and of a macrosomic fetus. High lateral implantation was associated with low Apgar scores. PMID- 17080096 TI - Extremely long hospitalizations of newborns in the United States: data, descriptions, dilemmas. AB - PROBLEM: Neonatal and pediatric nurses and physicians care for newborn children who have been saved by technological support but who then spend extremely long periods of time in the hospital, perhaps never being able to be discharged to home. There has been little research identifying newborns who are too sick to be discharged from the health care setting and rare reports of staff or parental response to these long-term hospitalizations. PURPOSE: This study provides both the numerical data and description of acutely, chronically ill newborn children whose illnesses caused hospitalizations for greater than 6-months (179 days) in the US. METHODOLOGY: Method triangulation using a national data set (HCUP-KID 2003), a researcher created LONGTERM survey, and a qualitative question was used to identify pathologies associated with newborn length of stays greater than 6 months. Neonatal nurses and physicians provided descriptions of children spending at least 6 months in the hospital, including anecdotal reports of caring for those children. RESULTS: The national H-CUP data set identified 680 infants staying 6 months or longer in the hospital during 2003. Four hundred and twenty two providers submitted LONGTERM surveys describing these infants, with 228 first hand reports on how it felt to care for children with hospital stays between 6 months and 6 years. Extreme prematurity, respiratory distress and necrotizing enterocolitis contributed to the extremely long hospital stays. Nurse and physician participants felt that extremely long hospital stays were often due to situations in which parents or colleagues were insisting upon continued futile treatment. PMID- 17080097 TI - Modern peritoneal dialysis: concepts and approaches. PMID- 17080098 TI - Glucose sparing in peritoneal dialysis: implications and metrics. AB - Glucose sparing is a central component of modern peritoneal dialysis therapy, necessary to reduce the trade-offs of the therapy in terms of membrane protection and alleviation of systemic consequences. We present a detailed exploration of a metric for this approach in the form of the index of ultrafiltration efficiency, that we propose as a guide for designing therapeutic regimens and for the development of new dialysis solutions. Based on insights from exploration of ultrafiltration efficiency with various dialysis solutions, we propose an approach to optimizing dialysis prescription. PMID- 17080099 TI - New insight of amino acid-based dialysis solutions. AB - Malnutrition is a major complication of peritoneal dialysis (PD) and is associated with increased morbidity and mortality. Daily losses of proteins and amino acids (AAs) into dialysate contribute to this problem. Previous metabolic balance study demonstrated that treatment with 1.1% AA-based dialysis solution is safe and may improve protein malnutrition in continuous ambulatory peritoneal dialysis (CAPD) patients ingesting low protein intake. Other prospective studies also showed that AA solution can provide nutritional benefit for malnourished PD patients resulting in a significant improvement in some biochemical and/or anthropometric nutritional parameters. However, there are other studies showing no particular improvement in nutritional parameters after long-term use of AA solution. This may be related to the differences in the study design, sample size, methods used to assess nutritional status, and other factors such as dietary intake and comorbidities of study subjects. Published data will be reviewed to further emphasize the nutritional benefit of long-term use of AA solution in malnourished PD patients along with a brief discussion on the various reasons that may partly explain the different study results. We will also present the results of a longitudinal observational study evaluating changes in nutritional parameters following use of one exchange of 1.1% AA solution in malnourished Korean PD patients. A significant improvement of somatic protein status such as lean body mass (LBM) and hand grip strength was observed. No significant change in serum albumin level was noted. Patients with a positive estimated coefficient for LBM in the fitted regression model to the repeated observations over 1 year were classified as responders and patients with neutral or negative coefficient were considered as non-responders. Thirty-one out of 43 malnourished patients (72%) showed nutritional benefit based on the change of LBM. Hand grip strength and back lift strength were significantly higher in responders at baseline. Other baseline parameters did not differ between the two groups. PMID- 17080100 TI - Management of hyperlipidemia in patients on peritoneal dialysis: current approaches. PMID- 17080101 TI - Structural requirements for a successful chronic peritoneal dialysis program. AB - Recently, there has been a decline in chronic peritoneal dialysis (CPD) utilization in several countries. And, in these countries, the percent of patients with end-sage renal disease maintained on CPD is less than nephrologists think is appropriate. The reasons for these problems are uncertain, but it is likely that difficulties with the structural organization of CPD facilities play a contributory role. This paper discusses the structural requirements for a successful CPD program, focusing attention on the following domains: (1) adequate chronic kidney disease education, (2) provision and support of physician training in the principles and practice of CPD, (3) adequate size and organization of CPD centers, (4) development of appropriate support systems within the CPD facility, and (5) development of appropriate continuous quality improvement programs to monitor a variety of domains, including adequacy of dialysis, peritonitis rates, catheter infections and problems, psychosocial status of patients, etc. PMID- 17080102 TI - French peritoneal dialysis registry (RDPLF): outline and main results. AB - The Registre de Dialyse Peritoneale de Langue Francaise (RDPLF Registry) is a non profit association that has been set up to assist physicians and nurses in evaluating their practical experience and results regarding peritoneal dialysis (PD). Five French-speaking and two Spanish-speaking countries have participated in this initiative (which includes 21 000 patients). In France, 82% of all PD patients are included in the registry and the main results for the period from 1995 to January 2006 form the basis of this report: of 11 744 incident patients with a median age of 71 years, 21.5% were over 80 years of age and 56% were not able to perform PD treatment at home without assistance. Eighty-six percent of the latter group received external assistance from a private nurse and 14% were aided by their family. The overall average rate of peritonitis was one episode every 29 months. The probability of being peritonitis-free appeared to be better for patients on automated PD (59.4% at 2 year) than for those on continuous ambulatory PD (55.3%), but this finding requires further validation. The average waiting time before transplantation was about 2 years. In patients who had undergone transplantation, the peritonitis rate was one episode per 42 months before transplantation compared to one episode per 29 months for patients who had not received a transplant. Eighty-three percent of patients had a hemoglobin level greater than 11 g%. Catheter survival was 92% at 2 years post-insertion and 85% at 5 years, with 94% being implanted by experienced surgeons. In conclusion, the RDPLF results demonstrate that PD may be successfully prescribed for older patients who receive assistance either from their family or from a nurse. Further, a larger number of younger patients should also be prescribed this technique in France. Patients eligible for transplantation and on short-term PD have the lowest risk of developing peritonitis; PD before transplantation may help prolong residual renal function, and initial treatment by PD may also help to preserve vascular access for the future. PMID- 17080103 TI - Nosogogy: when the learner is a patient with chronic renal failure. AB - Patient education approaches are currently derived from a biomedical 'acute' model characterized by the sequence of health, disease, and recovery resulting from our professional intervention. Unfortunately, this model proves to be totally inadequate when applied to a chronic disease such as kidney failure. Our patients never fully regain their health and may continue to worsen under our care, even after many state-of-the-art treatments. The solution is represented in acquiring a new professional identity, shifting from the 'biomedical' acute model to a 'bio-psycho-social-educational model'. Within this model, a Therapeutic Education approach in predialysis has been proven to provide both short- and long term positive results for renal patients. There is a tremendous difference between the learning processes in children and adults and two different sciences have already been described. 'Pedagogy' deals with child learning and 'Andragogy' with adult learning. Nevertheless, when the learner is a patient with a chronic disease, we believe that new considerations must be taken into account. We propose to create a novel science and to call it 'Nosogogy', derived from the ancient Greek word (see text), meaning 'disease'. Nosogogy could be defined as the science of teaching adults affected by chronic disease. The new educator is someone deeply involved in renal care who knows and understands the characteristic conflicts and dynamics that arise in the renal patient, and possesses adequate communication skills to deal with him. In our experience, we prefer to have educational sessions run by nephrologists and nurses who have great experience in the field. PMID- 17080104 TI - Patient re-training in peritoneal dialysis: why and when it is needed. AB - The aim of this multicenter, quantitative, observational study was to analyze compliance and re-training needs of patients on peritoneal dialysis (PD) through the assessment of patient knowledge (with a Patient Questionnaire; phase 1) and patient behavior (home visit with a Score Card; phase 2). A total of 353 patients from 11 Italian centers participated in the first phase and 191 patients from nine centers in the second phase. Overall, 66% of questions on the Patient Questionnaire were answered correctly. Correct answers were more frequent in females than males, in patients under 55 years of age, and in those with higher education. The lowest rate of correct answers involved questions related to diet and physical activity (67% and 51%, respectively). Data collected during the home visit showed that 25% of patients were partially compliant with their drug therapy. Twenty-three percent of patients were non-compliant with the exchange protocol procedures, with a significant association between compliance and the incidence of peritonitis, and 11% were non-compliant with the exit-site protocol procedures without a statistically significant correlation to peritonitis. By combining the two evaluations, we found that approximately one-third (29%) of patients needed reinforcement of knowledge and ability to correctly perform PD as related to infection control and 27% for the correct use of drugs. Looking at the combined evaluation of infection control and drug use, results showed that 47% of patients needed re-training. This need for re-training was greater for younger patients (less than 55 years old), patients with lower education degree and patients in the early or late phase of PD therapy (less than 18 months or more than 36 months). Gender and degree of autonomy had no effect on the need for re training. PMID- 17080105 TI - Patient and technique survival on peritoneal dialysis in patients with failed renal allograft: a case-control study. AB - Failed renal allograft is becoming one of the most frequent causes of dialysis initiation in countries with developed transplant programs. The majority of patients initiate hemodialysis (HD) as their next renal replacement modality and concerns about the success of peritoneal dialysis (PD) in this patient population has been expressed. We evaluated patient and technique outcome in a cohort of 494 patients in the United States who initiated PD after a failed renal allograft in the years 2000-2003, and compared the outcomes to those of two case-matched groups: patients new to dialysis or patients transferred from HD who started PD during the same period. Patients starting PD after a failed allograft had patient survival and technique survival similar to case-matched controls. Transplantation was lower in patients with failed allograft than controls. The high success of PD in patients with failed allograft suggests that it is beneficial to utilize this modality more frequently in this patient group than current practice. PMID- 17080106 TI - Place of genotyping and phenotyping in understanding and potentially modifying outcomes in peritoneal dialysis patients. AB - With the landmark publication of the human genome sequence and its subsequent division into haplotype blocks, the characterization of genetic variations is becoming a feasible approach to study both the pathophysiology and risk factors of complex traits. A number of strategies are available today for identifying candidate genes or polymorphisms associated with pertinent phenotypes. For Mendelian diseases with high penetrance owing to mutations in a single gene, such as polycystic kidney disease, linkage studies have been very successful in mapping the disease loci owing to the availability of families with multiple affected members. In contrast to monogenic conditions, complex diseases such as end-stage renal disease (ESRD) and complex traits such as individual variations in membrane transport and complications during the course of peritoneal dialysis (PD) therapy have a number of competing determinants and inhibitors, both genetic and environmental. Current results reflect this complexity, with few studies showing a large effect of any single risk factor on survival or outcome on PD. However, these studies have so far been small (less than 500 patients) and have not utilized bioinformatics or novel technologies (e.g., multiplex genotyping equipment). In the following review, we outline current approaches for using genetic data in clinical studies as well as highlight some of the most promising results in ESRD patients, particularly those on PD. PMID- 17080107 TI - Peritoneal dialysis in the US: evaluation of outcomes in contemporary cohorts. AB - Secular trends in dialysis therapy delivery require a frequent re-examination of outcomes in patients on renal replacement modalities. We examined four large cohorts of patients initiating peritoneal dialysis (PD) in 2000-2003 (total of >40 000 patients) to ascertain trends in patient outcomes, technique success, and predictors of both parameters of interest. Age, end-stage renal disease vintage, and diabetes were clear predictors of patient survival. Technique success was higher in patients on automated PD than in patients on continuous ambulatory PD. Center size was a powerful predictor of technique success. We conclude that the current state of PD in the United States is characterized by improving patient outcomes, higher technique success, and a predominance of use of cycler-based therapy. Several opportunities for improving technique success amenable to practice interventions have been identified. PMID- 17080108 TI - Selected best demonstrated practices in peritoneal dialysis access. AB - Many burdensome interventions that adversely affect the utilization of peritoneal dialysis as renal replacement therapy and patient satisfaction with this treatment modality can be avoided by early peritoneal access placement with embedded catheters, implantation techniques that preempt common catheter complications, and the use of access devices that provide flexibility in exit site location. Catheter embedding consists of subcutaneously burying the external limb of the catheter tubing at the time of the insertion procedure. Interval exteriorization of the catheter is performed when dialysis is needed. Earlier commitment by patients to peritoneal dialysis can be achieved by elimination of catheter maintenance until dialysis is necessary. Catheter embedding is a practical strategy to avoid temporary hemodialysis with vascular catheters and reduces stress on operating room access by allowing more efficient scheduling as non-urgent procedures. Laparoscopic catheter placement enables proactive techniques not available to other conventional insertion methods. These techniques include rectus sheath tunneling to prevent catheter tip migration, selective prophylactic omentopexy to prevent omental entrapment, selective resection of epiploic appendages to prevent catheter obstruction, adhesiolysis to eliminate compartmentalization, and diagnosis and simultaneous repair of previously undiagnosed abdominal wall hernias. Both standard and extended 2-piece catheter systems are necessary to customize the peritoneal access to a variety of body configurations. Catheters should be able to produce lower abdominal, mid abdominal, upper abdominal, and upper chest exit site locations that facilitate management by the patient without sacrificing deep pelvic position of the catheter tip or resulting in excessive tubing stress during passage through the abdominal wall. PMID- 17080109 TI - Mortality studies comparing peritoneal dialysis and hemodialysis: what do they tell us? AB - Several recent large-scale epidemiological studies comparing mortality among end stage renal disease (ESRD) patients receiving hemodialysis (HD) versus peritoneal dialysis (PD) show conflicting results. In this paper, we undertake a critical review of these studies. Our goal is to determine if there are any consistent trends in outcomes between HD and PD within select subgroups of patients once methodological differences have been accounted for. A total of six large-scale registry studies and three prospective cohort studies conducted in the United States (US), Canada, Denmark, and the Netherlands were reviewed. Summary findings from these studies are presented for comparative purposes. Additional summary analyses based on previously reported data on 398 940 incident US Medicare patients are included for the purpose of comparing results from this population of patients to those of the other select studies when similar methods of analysis are applied. Results are summarized in terms of the relative risk of death for PD versus HD (RR[PD:HD]). Differences in results between the nine studies can be attributed to the degree of case-mix adjustment carried out and to the use of different subgroups when comparing mortality between HD and PD. When these differences are accounted for, we found a remarkable degree of synergism in results between the registry studies and, to a lesser degree, the prospective cohort studies. PD was generally found to be associated with equal or better survival among non-diabetic patients and younger diabetic patients in all four countries. However, among older diabetic patients, results varied by country. The Canadian and Danish registries showed no difference in survival between PD and HD among older diabetics while in the US, HD was associated with better survival for diabetics aged 45 and older. All studies show a time-dependent trend in the RR of death with PD generally associated with equivalent or better survival during the first year or two of dialysis. However, results on longer-term survival varied according to study and to different subgroups within studies. Subgroup analyses in the prospective cohort studies were limited by small numbers of patients resulting in highly varied and somewhat controversial results when compared to the larger registry-based studies. Based on our review of recent publications and additional analyses of US Medicare data, we conclude that overall patient survival is similar for PD and HD but that important differences do exist within select subgroups of patients, particularly those subgroups defined by age and the presence or absence of diabetes. PMID- 17080110 TI - Use of the embedded peritoneal dialysis catheter: experience and results from a North American Center. AB - Since 2000, the Ottawa Hospital Home Dialysis Program has used a variation on the embedded peritoneal dialysis catheter technique described by Moncrief et al. In this paper, we describe our approach to placement of peritoneal access and report our experience with 304 embedded catheters placed between January 2000 and December 2003. We review the advantages and disadvantages of this technique and describe factors that have been important to the success of our program. PMID- 17080111 TI - Prevention of infectious complications in peritoneal dialysis: best demonstrated practices. AB - Peritoneal dialysis (PD) related infections continue to be a serious complication for PD patients. Peritonitis can be associated with pain, hospitalization and catheter loss as well as a risk of death. Peritonitis risk is not evenly spread across the PD population or programs. Very low rates of peritonitis in a program are possible if close attention is paid to the causes of peritonitis and protocols implemented to reduce the risk of infection. Protocols to decrease infection risk in PD patients include proper catheter placement, exit-site care that includes Staphylococcus aureus prophylaxis, careful training of patients with periodic retraining, treatment of contamination, and prevention of procedure related and fungal peritonitis. Extensive data have been published on the use of antibiotic prophylaxis to prevent exit site infections. There are fewer data on training methods of patients to prevent infection risk. Quality improvement programs with continuous monitoring of infections, both of the catheter exit site and peritonitis, are important to decrease the PD related infections in PD programs. Continuous review of every episode of infection to determine the root cause of the event should be routine in PD programs. Further research is needed examining approaches to decrease infection risk. PMID- 17080112 TI - Microbiology and outcomes of peritonitis in North America. AB - A proper understanding of the patterns of occurrence of infectious complications in patients on peritoneal dialysis (PD) is crucial to lay the foundation for interventions directed at the major causes and predisposing factors. The present work is an initial report of a survey based on a large representative database of the infectious complications of PD. Gram-positive organisms accounted for 62% of peritonitis episodes in the US and 61% of these episodes in Canada. Gram-negative organisms accounted for 20.5% of episodes in the US and 23.6% of episodes in Canada. The peritoneal catheter was removed in 18% of the episodes in the US and 16% in Canada. Less than 4% of the episodes resulted in death. Gram-positive organisms accounted for the majority of exit-site infections in the US (69%) and Canada (76%). Gram-negative organisms accounted for a third of exit-site infections and among Gram-negative organisms Pseudomonas was the overwhelmingly dominant organism. The microbiology of exit-site infections revealed the expected representation of Gram-positive organisms. An important finding, however, is the significantly high contribution of Gram-negative organisms. Our findings can inform future guideline development and suggest that similar endeavors be undertaken in other parts of the world. PMID- 17080113 TI - Impact of new dialysis solutions on peritonitis rates. AB - Peritonitis remains a major cause of morbidity among patients on peritoneal dialysis (PD), yet there is little information about the effect of new biocompatible dialysis solutions on peritonitis rates and treatment. In our unit, information on each peritonitis episode is prospectively collected. Since 2003, bicarbonate/lactate dialysate has been gradually introduced for new patients and for patients experiencing abdominal pain with conventional lactate solutions. From 2002 to 2005, data from 121 episodes of peritonitis (71 automated PD and 50 continuous ambulatory PD) were analyzed; 107 episodes occurred in patients using standard lactate dialysate and 14 episodes in patients using bicarbonate/lactate solution. Patients using bicarbonate/lactate had a significantly lower peritonitis rate of 1 per 52.5 patient-months compared to those using standard lactate dialysate (1 per 26.9 patient-months) (P=0.0179). Response to treatment, however, was not affected by the type of dialysate; cure rates (71.4 and 69.1%, respectively) and recurrence rates (21.4 and 15.8%, respectively) were not significantly different. Catheter removal was required in three (21.4%) patients using bicarbonate/lactate and 23 (22.4%) patients using lactate solution. Use of biocompatible dialysate appears to reduce the peritonitis rate by 50%, although this has to be confirmed in a randomized study. The type of dialysate, on the other hand, does not affect response to treatment. PMID- 17080114 TI - Place of peritoneal dialysis in the management of treatment-resistant congestive heart failure. PMID- 17080115 TI - The role of peritoneal dialysis in the management of treatment-resistant congestive heart failure: A European perspective. AB - Patients with congestive heart failure (CHF) resistant to conventional treatment have a poor prognosis. Extracorporeal ultrafiltration (UF) appears to be the therapy of choice for short-term management of such patients with severe fluid overload, whereas peritoneal dialysis (PD) may be the therapy of choice for the long-term treatment. Fluid removal results in reduction of plasma volume, improvement of hyponatremia, reduction in pulmonary capillary wedge pressure, improvement of New York Heart Association functional heart failure class, improvement of functional rehabilitation and quality of life, reduction of hospitalizations and readmissions, as well as improvement in diuretic responsiveness. Whether extracorporeal UF and/or PD modifies the survival rate of patients with refractory CHF needs to be determined in prospective randomized controlled trials. PMID- 17080116 TI - Mitigating peritoneal membrane characteristics in modern peritoneal dialysis therapy. AB - Membrane function at the start of peritoneal dialysis (PD) treatment, measured as solute transport rate and ultrafiltration capacity, varies considerably between individuals. Although this can be correlated to clinical factors such as age and body habitus, this accounts for little of the variance seen. It is increasingly clear, however, that this variability in membrane function does impact on clinical outcomes. Specifically, high solute transport increases mortality risk, independent of other known factors such as age, comorbidity, and residual renal function. High solute transport causes earlier loss of the osmotic gradient when a low molecular weight osmolyte such as glucose is used. This will result in an earlier and lower peak in the ultrafiltration achieved combined with a higher fluid absorption rate once the osmotic gradient is lost. It is therefore quite plausible that the worse clinical outcomes associated with high transport reflect less good ultrafiltration, although other explanations must be considered, including higher peritoneal protein losses and a possible association with systemic inflammation. Strategies now exist to mitigate the effects of high transport on fluid removal. These include optimization of the short dwell lengths using automated PD (APD) combined with icodextrin which will result in sustained ultrafiltration and thus prevention of reabsorption in the long dwell. Survival analysis of APD patients, especially in cohorts in which icodextrin has been used, would suggest that high transport status is not a risk factor, although some of these data are only preliminary. In contrast, low ultrafiltration capacity of the membrane seems to be more important in these patients, especially if anuric. Here the best strategy would seem to be prevention as patients who develop low ultrafiltration capacity are not easily treated on PD. Avoiding excessive hypertonic glucose exposure and preserving residual renal function offers the best available approach. PMID- 17080117 TI - Profiles of automated peritoneal dialysis prescriptions in the US 1997-2003. AB - Automated peritoneal dialysis (PD) is the dominant mode of delivery of PD in the US. Information about actual prescribing patterns has been limited. The present study examines cycler prescription use in large cohorts during the years 1997, 2000, and 2003. We observed trends consisting of increasing fill volumes, increasing time on cycler, lowering of the number of cycles, and a shift in the reason for utilization of tidal therapy. Monitoring of practice patterns is beneficial in identifying opportunities for practice enhancement. The findings of the present survey demonstrate trends that approach standard recommendations of the benefits of increasing fill volume whereas paying close attention to dwell time as a balance between cycle number and therapy duration. Furthermore, this survey shows that the cycler represents a flexible method to personalize PD therapy. PMID- 17080118 TI - Tidal PD: its role in the current practice of peritoneal dialysis. AB - The role of tidal peritoneal dialysis (TPD) has been the subject of several studies over the past 30 years. The use of the newest generation of cyclers combined with the increasing number of chronic peritoneal dialysis (CPD) patients being maintained on cycler therapy has stimulated a reexamination of the role of TPD in the care of CPD patients. Several studies over the past decade have examined solute clearances with TPD in patients. These studies suggest that TPD does not result in an increase in clearances when compared to conventional intermittent peritoneal dialysis (IPD). TPD is now primarily used for comfort in patients who experience pain at the start of inflow and/or at the end of outflow. In TPD, the presence of at least some fluid in the abdomen during the exchanges generally eliminates these episodes of pain. It has recently been suggested that accurate assessment of drain and fill phases during automated PD may be helpful in redefining a role for TPD in CPD patients. If the 'slow' drainage time can be kept to a minimum, then it is possible that the efficiency of PD could be enhanced. Defining the critical volume and then optimizing the TPD regimen could perhaps increase the clearances noted with TPD. PMID- 17080119 TI - The role of tidal peritoneal dialysis in modern practice: A European perspective. AB - Tidal peritoneal dialysis (TPD) has been introduced to optimize adequacy of peritoneal dialysis (PD). Early studies reported similar or even better small solute clearances with TPD than those achieved with continuous ambulatory peritoneal dialysis or continuous cyclic peritoneal dialysis. However, in many studies treatment volumes were much higher during TPD compared with other PD modalities. Based on current evidence, TPD provides no advantage of increased small solute clearances, middle molecule clearances, or peritoneal ultrafiltration as compared to non-tidal automated peritoneal dialysis (APD) when dialysate flow is kept constant. However, TPD reduces drainage pain and nightly alarms during cycler treatment. Tidal volume should be kept as high as possible in these patients, especially in those with low average peritoneal transport rates. Based on theoretical considerations and little evidence, TPD could provide better clearances than conventional APD when a very high dialysate flow (>or=5 l/h) is used. Such dialysate flow rates are not routinely prescribed in home APD patients. However, they may be interesting for in-center PD patients. One randomized crossover trial reported higher small solute clearances with TPD compared to non-tidal APD in patients with acute renal failure. TPD is also the preferred treatment modality in patients with ascites as it allows a controlled outflow of fluid from the peritoneal cavity. Newer treatment modalities, for example, continuous flow PD, may be interesting alternatives in an effort to increase efficacy of PD in the future. However, because such treatment regimens are expensive and elaborate they have not been established for routine use until now. PMID- 17080153 TI - Organ transplantation goes to the movies. PMID- 17080154 TI - Secondary oxalosis due to excess vitamin C intake. PMID- 17080155 TI - Sclerosing encapsulating peritonitis in a 47-year-old woman. PMID- 17080156 TI - A crucial nephron segment in acid-base and electrolyte transport: the connecting tubule. AB - The cortical distal nephron is a heterogenous structure where the fine regulation of electrolyte and water balance takes place. Among the other segments, previous reports have emphasized the importance of the connecting tubule in sodium and potassium renal handling. Kovacikova et al. report that the connecting tubule is also the major segment in electrogenic urinary acidification, thus reinforcing the central role of this segment in overall electrolyte transport. PMID- 17080157 TI - Focal segmental glomerulosclerosis: cellular variant and beyond. AB - The entity of focal segmental glomerulosclerosis (FSGS), oddly, includes several distinct changes involving glomeruli that need not be focal, segmental, or even sclerotic. It is fitting to rethink our nosological approach to FSGS, which has focused on descriptive morphological entities. Rather, we should consider them as 'podocytopathies' - diseases with an etiological commonality involving injury to the podocyte. PMID- 17080158 TI - Adiponectin versus angiotensin II: Key pathological role of their misbalance. AB - Kurata et al. report that an angiotensin II type-1 receptor blocker (ARB) inhibits obesity-induced hypoadiponectinemia in rats through inhibition of reactive oxygen species generation. Taken together with the recently reported functional interplay between adiponectin and angiotensin II, this suggests that adiponectin probably mediates tissue-protective effects of ARBs in obesity and metabolic syndrome. PMID- 17080161 TI - Longer treatment time and slower ultrafiltration in hemodialysis: associations with reduced mortality in the Dialysis Outcomes and Practice Patterns Study. PMID- 17080162 TI - Calculation of glomerular filtration rate using serum cystatin C in kidney transplant recipients. PMID- 17080165 TI - Renin-angiotensin inhibitors in stage 4 chronic kidney disease. PMID- 17080166 TI - Is inflammation the missing link between low fat mass and low survival in hemodialysis patients? PMID- 17080167 TI - Urotensin in ESRD: not so surprising. PMID- 17080169 TI - The evolution of the PCPT from clinical to molecular analyses. PMID- 17080170 TI - Cancer in the developing world: can we avoid the disaster? PMID- 17080171 TI - Can thalidomide improve outcome in patients with multiple myeloma? PMID- 17080172 TI - Poor correlation between physician and patient assessment of quality of life in palliative care. PMID- 17080173 TI - Rituximab and CHOP-like chemotherapy in good-prognosis diffuse large-B-cell lymphoma. PMID- 17080174 TI - Can the utility of secondary cytoreduction for ovarian cancer be reliably established? PMID- 17080175 TI - Focusing on physical function limitations in elderly women surviving cancer: any opportunity for improvement? PMID- 17080176 TI - Does tumor mutational status correlate with clinical response to imatinib? PMID- 17080177 TI - Stabilization of disease in patients with metastatic renal cell carcinoma using sorafenib. PMID- 17080178 TI - Ovarian cancer: a focus on management of recurrent disease. AB - Surgery and chemotherapy form the cornerstone of the treatment for ovarian cancer. Currently, the standard of care for primary ovarian cancer is platinum and taxane-based therapy. Even among women with advanced and suboptimal disease (i.e. tumors greater than 1 cm) following surgery, the clinical efficacy of chemotherapy is noteworthy. Despite the favorable response characteristics, however, most women with advanced-stage ovarian cancer will relapse, including about 50% of women who have no evidence of disease after primary therapy. A multitude of treatment options are available at the time of recurrence, but there is no clear consensus about how these patients should be managed. Options include surgery, chemotherapy, hormones, and sometimes, radiation therapy. The sequence, combinations of treatment, and manner in which any or all of these options should be employed in an individual patient, which heretofore have not been standardized, are the subjects of ongoing clinical investigations. PMID- 17080179 TI - Stress and breast cancer: a systematic update on the current knowledge. AB - A vast body of research has been carried out to examine the relationship between psychological stress and the risk of breast cancer. Previous reviews on this issue have mainly focused on stressful life events and have included both prospective and retrospective studies. The results from these reviews have revealed conflicting data. We evaluate whether stressful life events, work related stress, or perceived global stress are differentially associated with breast cancer incidence and breast cancer relapse in prospective studies. Systematic and explicit methods were used to identify, select, and critically appraise relevant studies. The substantial variability in the manner in which stress was conceptualized and measured did not allow for the calculation of a quantitative summary estimate for the association between stress and breast cancer. Despite the heterogeneity in the results obtained, it is concluded that stress does not seem to increase the risk of breast cancer incidence. Whether stress affects the progression of breast cancer is still unclear. Studies with more thorough adjustment for confounding factors and larger studies on stress and breast cancer relapse are required to address this issue. PMID- 17080180 TI - Molecular classification of breast cancer: implications for selection of adjuvant chemotherapy. AB - Adjuvant chemotherapy improves survival of patients with stage I-III breast cancer but it is being increasingly recognized that the benefit is not equal for all patients. Molecular characteristics of the cancer affect sensitivity to chemotherapy. In general, estrogen-receptor-negative disease is more sensitive to chemotherapy than estrogren-receptor-positive disease. Large-scale genomic analyses of breast cancer suggest that further molecular subsets may exist within the categories defined by hormone receptor status. It is hoped that the new molecular classification schemes might improve patient selection for therapy. Before any new molecular classification (or predictive test) is adopted for routine clinical use, however, several criteria need to be met. There must be an agreed and reproducible method by which to assign molecular class to a new case. Cancers that belong to different molecular classes must show differences in disease outcome and treatment efficacy that affect management and treatment selection. Also desirable are results from prospective clinical trials that demonstrate improved patient outcome when the new test is used in decision making, compared with the current standard of care. This Review describes the current limitations and future promises of gene-expression-based molecular classification of breast cancer and how it might impact on selection of adjuvant therapy for individual patients. PMID- 17080181 TI - Interferon beta adenoviral gene therapy in a patient with ovarian cancer. AB - Background A 47-year-old woman with a history of ovarian cancer and a 6-year disease-free remission presented with dyspnea and increased abdominal girth. The patient was found to have ascites and a large left pleural effusion, both of which contained malignant cells consistent with recurrent ovarian cancer. Her disease progressed despite treatment with chemotherapeutic and hormonal agents. She was then enrolled in a phase I clinical trial of adenoviral-mediated interferon beta gene therapy. Investigations Abdominal and chest CT scans, 2 [(18)F]fluoro-2-deoxyglucose PET scan, viral cultures, interferon cytokine analysis, immunophenotyping, and tumor cytotoxicity analyses. Diagnosis Stage IV ovarian cancer with malignant ascites and pleural effusion. Management Tunneled pleural catheter and intrapleural adenoviral-mediated interferon beta gene therapy. PMID- 17080182 TI - Effect of gene delivery of NOS isoforms on intimal hyperplasia and endothelial regeneration after balloon injury. AB - Endothelial cell loss is a critical event in the pathological repair of the injured blood vessel. Impaired endothelial function results in reduced production of key vascular mediators such as nitric oxide (NO) within the vessel wall leading to enhanced smooth muscle cell proliferation and migration and ultimately intimal hyperplasia. The aim of the present study was to directly compare the effects of adenoviral-mediated gene delivery of two nitric oxide synthase (NOS) isoforms, eNOS and iNOS on endothelial regeneration and intimal hyperplasia following endothelial injury in the rabbit carotid artery. The right carotid arteries of male New Zealand white rabbits were denuded by passing a 3French Fogarty balloon catheter along the artery three times. In all, 1 x 10(9) PFU of adenoviral(Ad)eNOS, AdiNOS or Adbeta-galactosidase (Adbeta-Gal) was then delivered intraluminally and allowed to dwell for 20 min. Transgene expression was sought after 3 days by immunohistochemistry and at 7 days by quantitative reverse transcriptase PCR. The effect on intimal hyperplasia was sought using histological staining after 14 days. Evans blue staining was used to determine the effect on endothelial regeneration. eNOS and iNOS expression was detected in transduced arteries. Neointima/media ratios were significantly reduced in eNOS (0.07+/-0.044) and iNOS (0.087+/-0.086) transduced arteries compared with Adbeta Gal (0.332+/-0.14) transduced arteries (n=7). In addition, AdeNOS treatment (4.21+/-3.12% de-endothelialized area) enhanced endothelial regeneration compared to Adbeta-Gal treatment (10.05+/-4.98), while treatment with AdiNOS (25.17+/ 11.92) inhibited endothelial regeneration in the injured rabbit carotid artery (n=7-8). These results highlight the potential of NOS gene therapy, in particular, eNOS gene therapy as a potential therapeutic strategy for the prevention of restenosis after vascular injury. PMID- 17080183 TI - Relationship Between Religious Coping and Suicidal Behaviors Among African American Adolescents. AB - This study investigated whether hopelessness and depression were risk factors for suicidal thoughts and behaviors in African American adolescents and looked at whether religious participation and religious coping protected these students from suicidality. Participants were 212 African American high school students (133 females, 79 males). The results of multiple and logistic regression analyses found that hopelessness and depression were risk factors for suicidal ideation and attempts. Religious coping style was significantly related to suicidal behaviors: Self-directed coping was related to increased hopelessness, depression, and suicide attempts, and collaborative coping was related to increased reasons for living. Gender differences were found in symptoms of depression, religious coping style and religious participation. Results provide additional support for suicide interventions to target hopelessness and depressive symptoms and highlight the importance of examining the role of culturally salient variables, such as religious participation and religious coping style, when developing intervention programs for suicide. PMID- 17080184 TI - Can I quote you on that? AB - Just as a simple phrase can become convoluted in the childhood game of Telephone, research findings can be distorted in the lay press. Journalists and scientists themselves must share the responsibilities of better explaining and interpreting science in an accessible and meaningful context for nonspecialist readers. PMID- 17080185 TI - A man who has made his Marks on science. PMID- 17080186 TI - Response to Windahl et al. PMID- 17080187 TI - Drug discovery in jeopardy. AB - Despite striking advances in the biomedical sciences, the flow of new drugs has slowed to a trickle, impairing therapeutic advances as well as the commercial success of drug companies. Reduced productivity in the drug industry is caused mainly by corporate policies that discourage innovation. This is compounded by various consequences of mega-mergers, the obsession for blockbuster drugs, the shift of control of research from scientists to marketers, the need for fast sales growth, and the discontinuation of development compounds for nontechnical reasons. Lessons from the past indicate that these problems can be overcome, and herein, new and improved directions for drug discovery are suggested. PMID- 17080189 TI - Dysfunction of TGF-beta signaling in Alzheimer's disease. AB - Accumulation of beta-amyloid peptide (Abeta) in the brain is believed to trigger a complex and poorly understood pathologic reaction that results in the development of Alzheimer's disease (AD). Despite intensive study, there is no consensus as to how Abeta accumulation causes neurodegeneration in AD. In this issue of the JCI, Tesseur et al. report that the expression of TGF-beta type II receptor (TbetaRII) by neurons is reduced very early in the course of AD and that reduced TGF-beta signaling increased Abeta deposition and neurodegeneration in a mouse model of AD (see the related article beginning on page 3060). Intriguingly, reduced TGF-beta signaling in neuroblastoma cells resulted in neuritic dystrophy and increased levels of secreted Abeta. Collectively, these data suggest that dysfunction of the TGF-beta/TbetaRII signaling axis in the AD brain may accelerate Abeta deposition and neurodegeneration. PMID- 17080188 TI - Myasthenia gravis: past, present, and future. AB - Myasthenia gravis (MG) is an autoimmune syndrome caused by the failure of neuromuscular transmission, which results from the binding of autoantibodies to proteins involved in signaling at the neuromuscular junction (NMJ). These proteins include the nicotinic AChR or, less frequently, a muscle-specific tyrosine kinase (MuSK) involved in AChR clustering. Much is known about the mechanisms that maintain self tolerance and modulate anti-AChR Ab synthesis, AChR clustering, and AChR function as well as those that cause neuromuscular transmission failure upon Ab binding. This insight has led to the development of improved diagnostic methods and to the design of specific immunosuppressive or immunomodulatory treatments. PMID- 17080190 TI - Microglia: a cellular vehicle for CNS gene therapy. AB - Metachromatic leukodystrophy (MLD) is a lysosomal storage disease caused by deficiency of the enzyme arylsulfatase A (ARSA). MLD is characterized by progressive demyelination and neurological deficits. Treatment of MLD is still a challenge due to the fact that the blood-brain barrier is a major obstacle for most therapeutic substances. In this issue of the JCI, Biffi et al. report that genetically modified hematopoietic precursor cells transduced to overexpress ARSA and transplanted into mice with a targeted disruption of the murine Arsa gene (Arsa(-/-) mice) migrated into the CNS and cross-corrected brain ARSA deficiency (see the related article beginning on page 3070). Microglia served as a cellular vehicle to effectively deliver the enzyme to other brain cells while hepatocytes overexpressing ARSA increased plasma ARSA levels but failed to deliver ARSA into the CNS. PMID- 17080191 TI - Shaping the sperm head: an ER enzyme leaves its mark. AB - Lipid storage diseases are debilitating inherited metabolic disorders that stem from the absence of specific lysosomal enzymes that degrade selected lipids. Most characteristically, these disorders affect the nervous and the reticulo endothelial systems, with massive organomegaly resulting from the presence of engorged, lipid-laden macrophages. In this issue of the JCI, Yildiz et al. describe the role of the ER-resident enzyme beta-glucosidase 2 (GBA2) in mice (see the related article beginning on page 2985). Surprisingly, GBA2 deficiency leaves bile acid and cholesterol metabolism intact, instead causing lipid accumulation in the ER of testicular Sertoli cells, round-headed sperm (globozoospermia), and impaired male fertility. PMID- 17080192 TI - The paradoxical patent ductus arteriosus. AB - The ductus arteriosus (DA) is a vessel whose patency is required for fetal survival but is incompatible with postnatal life. Because of developmental insufficiency, the DA in preterm infants often fails to close in a condition known as patent DA (PDA). Although COX inhibitors can be used to close the PDA by lowering circulating prostaglandin levels, their effectiveness is correlated with birth weight, and severely premature infants often require surgical repair. Paradoxically, targeted deletion of COX pathway components in mice results in PDA. In this issue of the JCI, Yokoyama et al. describe dual roles for prostaglandins in DA development and closure, offering new insights into the mechanism of negative effects of COX inhibitors that may influence the treatment of severely premature infants with PDA and lead to improvement of their outcomes (see the related article beginning on page 3026). PMID- 17080193 TI - Role for IKK2 in muscle: waste not, want not. AB - Activation of transcription factor NF-kappaB, the major regulator of the inflammatory response, depends on the inhibitor of NF-kappaB kinase (IKK) complex, which is composed of 2 catalytic subunits, IKK1 and IKK2 (also known as IKKalpha and IKKbeta), and a regulatory subunit, IKKgamma (also known as NEMO). In this issue of the JCI, Mourkioti et al. show that muscle-specific disruption in mice of the gene encoding IKK2 prevents NF-kappaB activation in response to denervation or toxin-induced injury (see the related article beginning on page 2945). Importantly, this genetic manipulation prevents muscle wasting, thereby providing strong evidence in support of a major pathogenic role for inflammation in a variety of muscular dystrophies characterized by progressive muscle fiber degeneration. PMID- 17080194 TI - Local application of FTY720 to the lung abrogates experimental asthma by altering dendritic cell function. AB - Airway DCs play a crucial role in the pathogenesis of allergic asthma, and interfering with their function could constitute a novel form of therapy. The sphingosine 1-phosphate receptor agonist FTY720 is an oral immunosuppressant that retains lymphocytes in lymph nodes and spleen, thus preventing lymphocyte migration to inflammatory sites. The accompanying lymphopenia could be a serious side effect that would preclude the use of FTY720 as an antiasthmatic drug. Here we show in a murine asthma model that local application of FTY720 via inhalation prior to or during ongoing allergen challenge suppresses Th2-dependent eosinophilic airway inflammation and bronchial hyperresponsiveness without causing lymphopenia and T cell retention in the lymph nodes. Effectiveness of local treatment was achieved by inhibition of the migration of lung DCs to the mediastinal lymph nodes, which in turn inhibited the formation of allergen specific Th2 cells in lymph nodes. Also, FTY720-treated DCs were intrinsically less potent in activating naive and effector Th2 cells due to a reduced capacity to form stable interactions with T cells and thus to form an immunological synapse. These data support the concept that targeting the function of airway DCs with locally acting drugs is a powerful new strategy in the treatment of asthma. PMID- 17080195 TI - Targeted ablation of IKK2 improves skeletal muscle strength, maintains mass, and promotes regeneration. AB - NF-kappaB is a major pleiotropic transcription factor modulating immune, inflammatory, cell survival, and proliferative responses, yet the relevance of NF kappaB signaling in muscle physiology and disease is less well documented. Here we show that muscle-restricted NF-kappaB inhibition in mice, through targeted deletion of the activating kinase inhibitor of NF-kappaB kinase 2 (IKK2), shifted muscle fiber distribution and improved muscle force. In response to denervation, IKK2 depletion protected against atrophy, maintaining fiber type, size, and strength, increasing protein synthesis, and decreasing protein degradation. IKK2 depleted mice with a muscle-specific transgene expressing a local Igf-1 isoform (mIgf-1) showed enhanced protection against muscle atrophy. In response to muscle damage, IKK2 depletion facilitated skeletal muscle regeneration through enhanced satellite cell activation and reduced fibrosis. Our results establish IKK2/NF kappaB signaling as an important modulator of muscle homeostasis and suggest a combined role for IKK inhibitors and growth factors in the therapy of muscle diseases. PMID- 17080196 TI - Mutation of beta-glucosidase 2 causes glycolipid storage disease and impaired male fertility. AB - beta-Glucosidase 2 (GBA2) is a resident enzyme of the endoplasmic reticulum thought to play a role in the metabolism of bile acid-glucose conjugates. To gain insight into the biological function of this enzyme and its substrates, we generated mice deficient in GBA2 and found that these animals had normal bile acid metabolism. Knockout males exhibited impaired fertility. Microscopic examination of sperm revealed large round heads (globozoospermia), abnormal acrosomes, and defective mobility. Glycolipids, identified as glucosylceramides by mass spectrometry, accumulated in the testes, brains, and livers of the knockout mice but did not cause obvious neurological symptoms, organomegaly, or a reduction in lifespan. Recombinant GBA2 hydrolyzed glucosylceramide to glucose and ceramide; the same reaction catalyzed by the beta-glucosidase acid 1 (GBA1) defective in subjects with the Gaucher's form of lysosomal storage disease. We conclude that GBA2 is a glucosylceramidase whose loss causes accumulation of glycolipids and an endoplasmic reticulum storage disease. PMID- 17080197 TI - Secreted PCSK9 decreases the number of LDL receptors in hepatocytes and in livers of parabiotic mice. AB - Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a member of the proteinase K subfamily of subtilases that reduces the number of LDL receptors (LDLRs) in liver through an undefined posttranscriptional mechanism. We show that purified PCSK9 added to the medium of HepG2 cells reduces the number of cell surface LDLRs in a dose- and time-dependent manner. This activity was approximately 10-fold greater for a gain-of-function mutant, PCSK9(D374Y), that causes hypercholesterolemia. Binding and uptake of PCSK9 were largely dependent on the presence of LDLRs. Coimmunoprecipitation and ligand blotting studies indicated that PCSK9 and LDLR directly associate; both proteins colocalized to late endocytic compartments. Purified PCSK9 had no effect on cell-surface LDLRs in hepatocytes lacking autosomal recessive hypercholesterolemia (ARH), an adaptor protein required for endocytosis of the receptor. Transgenic mice overexpressing human PCSK9 in liver secreted large amounts of the protein into plasma, which increased plasma LDL cholesterol concentrations to levels similar to those of LDLR-knockout mice. To determine whether PCSK9 was active in plasma, transgenic PCSK9 mice were parabiosed with wild-type littermates. After parabiosis, secreted PCSK9 was transferred to the circulation of wild-type mice and reduced the number of hepatic LDLRs to nearly undetectable levels. We conclude that secreted PCSK9 associates with the LDLR and reduces hepatic LDLR protein levels. PMID- 17080198 TI - Chronic activation of the prostaglandin receptor EP4 promotes hyaluronan-mediated neointimal formation in the ductus arteriosus. AB - PGE, a potent vasodilator, plays a primary role in maintaining the patency of the ductus arteriosus (DA). Genetic disruption of the PGE-specific receptor EP4, however, paradoxically results in fatal patent DA (PDA) in mice. Here we demonstrate that EP4-mediated signals promote DA closure by hyaluronic acid mediated (HA-mediated) intimal cushion formation (ICF). Chronic EP4 stimulation by ONO-AE1-329, a selective EP4 agonist, significantly enhanced migration and HA production in rat DA smooth muscle cells. When HA production was inhibited, EP4 mediated migration was negated. Activation of EP4, adenylyl cyclase, and PKA all increased HA production and the level of HA synthase 2 (HAS2) transcripts. In immature rat DA explants, ICF was promoted by EP4/PKA stimuli. Furthermore, adenovirus-mediated Has2 gene transfer was sufficient to induce ICF in EP4 disrupted DA explants in which the intimal cushion had not formed. Accordingly, signals through EP4 have 2 essential roles in DA development, namely, vascular dilation and ICF. The latter would lead to luminal narrowing, helping adhesive occlusion and permanent closure of the vascular lumen. Our results imply that HA induction serves as an alternative therapeutic strategy for the treatment of PDA to the current one, i.e., inhibition of PGE signaling by cyclooxygenase inhibitors, which might delay PGE-mediated ICF in immature infants. PMID- 17080199 TI - Deficiency in neuronal TGF-beta signaling promotes neurodegeneration and Alzheimer's pathology. AB - Alzheimer's disease (AD) is characterized by progressive neurodegeneration and cerebral accumulation of the beta-amyloid peptide (Abeta), but it is unknown what makes neurons susceptible to degeneration. We report that the TGF-beta type II receptor (TbetaRII) is mainly expressed by neurons, and that TbetaRII levels are reduced in human AD brain and correlate with pathological hallmarks of the disease. Reducing neuronal TGF-beta signaling in mice resulted in age-dependent neurodegeneration and promoted Abeta accumulation and dendritic loss in a mouse model of AD. In cultured cells, reduced TGF-beta signaling caused neuronal degeneration and resulted in increased levels of secreted Abeta and beta secretase-cleaved soluble amyloid precursor protein. These results show that reduced neuronal TGF-beta signaling increases age-dependent neurodegeneration and AD-like disease in vivo. Increasing neuronal TGF-beta signaling may thus reduce neurodegeneration and be beneficial in AD. PMID- 17080200 TI - Gene therapy of metachromatic leukodystrophy reverses neurological damage and deficits in mice. AB - Metachromatic leukodystrophy (MLD) is a demyelinating lysosomal storage disorder for which new treatments are urgently needed. We previously showed that transplantation of gene-corrected hematopoietic stem progenitor cells (HSPCs) in presymptomatic myeloablated MLD mice prevented disease manifestations. Here we show that HSC gene therapy can reverse neurological deficits and neuropathological damage in affected mice, thus correcting an overt neurological disease. The efficacy of gene therapy was dependent on and proportional to arylsulfatase A (ARSA) overexpression in the microglia progeny of transplanted HSPCs. We demonstrate a widespread enzyme distribution from these cells through the CNS and a robust cross-correction of neurons and glia in vivo. Conversely, a peripheral source of enzyme, established by transplanting ARSA-overexpressing hepatocytes from transgenic donors, failed to effectively deliver the enzyme to the CNS. These results indicate that the recruitment of gene-modified, enzyme overexpressing microglia makes the enzyme bioavailable to the brain and makes therapeutic efficacy and disease correction attainable. Overall, our data provide a strong rationale for implementing HSPC gene therapy in MLD patients. PMID- 17080201 TI - Animal toxins as analgesics--an overview. AB - Pain is a multidimensional sensory experience, and multiple mechanisms are involved in the generation of pathophysiological nociceptive pain. Identification of the mechanisms and molecular components responsible for pain generation has not only advanced our understanding of pain and its control, but has also led to the selection of new targets for designing novel analgesic drugs. The high selectivity and specificity of animal toxins have enabled their use as potential therapeutics in the treatment of pain and candidates for the development of new analgesic drugs. This review focuses on the use of animal toxins for pain control and examines the possible analgesic mechanisms of these molecules. PMID- 17080202 TI - Cortistatin as a potential multistep therapeutic agent for inflammatory disorders. AB - The induction of immune tolerance is critical for the prevention of autoimmunity and the maintenance of immune homeostasis. The identification of factors involved in the maintenance or restoration of such tolerance has become the focus of new therapies for inflammatory and autoimmune diseases. Cortistatin, a recently discovered cyclic neuropeptide related to somatostatin, has emerged as a potential endogenous antiinflammatory factor based on its production by, as well as its binding to, immune cells. Thus, cortistatin has been found to downregulate the inflammatory response mediated by activated macrophages. The present work reviews various recent studies involving different experimental models of sepsis, rheumatoid arthritis and inflammatory bowel disease, demonstrating that cortistatin treatment offers great benefits at both the clinical and pathological levels. These include the downregulation of both inflammatory and Th1-mediated autoimmune disease components and the emergence of regulatory T cells (Treg) that suppress autoreactive T cells, both of which contribute to the restoration of immune tolerance. While many questions need to be resolved, cortistatin appears to be an exciting and promising candidate for the treatment of several chronic inflammatory diseases and autoimmune disorders. PMID- 17080203 TI - The role of galactosyltransferases in cell surface functions and in the immune system. AB - The immune system relies on cellular communication and often on the recognition of carbohydrates by mammalian lectins. Galactose (Gal)-containing structures are involved in both the innate and adaptive immune systems. Gal is a ligand for Gal/N-acetylgalactosmine (GalNAc) receptors and galectins, and is part of the scaffold structure that synthesizes oligosaccharide ligands for selectins, siglecs and other lectins of the immune system. Gal residues are added to glycoproteins and glycolipids by members of a large family of galactosyltransferases. The expression of many of these enzymes is regulated by the action of cytokines, and becomes altered in various disease states. Specific galactosyltransferases have been shown to control cell adhesion and leukocyte functions. Antibodies need to be galactosylated for normal function, and undergalactosylated immunoglobulin (Ig) is associated with rheumatoid arthritis, while Gal is lacking in the IgA of patients with IgA nephropathy. Interactions involving Gal play important roles in host defenses; they can also result in serious pathophysiology. Galactosyltransferases represent potential targets for the control of cell growth and apoptosis, inflammation and infections. PMID- 17080204 TI - The role of heparan sulfate in the generation of Abeta. AB - Alzheimer's disease is a fatal neurodegenerative disorder for which there are currently few treatments and no cure. Heparan sulfate, a heterogeneously sulfated glycosaminoglycan, has been identified as the first naturally occurring inhibitor of beta secretase, the rate-limiting step in the formation of Abeta, the peptide core of the amyloid plaques that cause Alzheimer's disease. Though heparan sulfate has frequently been implicated in the formation of fibrils, only fairly recently has its role as an inhibitor of beta secretase been recognized. This inhibitory activity is dependent on the structure and size of the heparan sulfate chain, with emphasis placed on the position of the sulfates. Heparan sulfate directly binds to beta secretase and causes a closed configuration of the catalytic site. Regulation of amyloid precursor protein (APP) beta secretase cleavage could occur at a number of cellular locations, including the Golgi complex, endosomal system and cell surface. Heparan sulfate also binds to APP and may sequester it away from beta secretase. These findings have led to the examination of heparan sulfate analogues, such as beta-secretase inhibitors, as a potential therapeutic approach to treat Alzheimer's disease. PMID- 17080205 TI - Spotlight on lamotrigine for depression. AB - Augmentation strategy is an option for patients who do not respond to one or two adequate trials of antidepressant therapy and are considered treatment-resistant. A number of drugs have been proposed for this indication, including lamotrigine, an anticonvulsant that is also licensed for the treatment of bipolar I disorder. Three retrospective chart reviews provided preliminary evidence of efficacy. However, other studies have been very small open-label studies, with the exception of one randomized, double-blind study. Although these studies indicate that lamotrigine may be effective as augmentation therapy in treatment-resistant depression, larger controlled studies are required to confirm these initial findings and to provide evidence for its use in this indication. PMID- 17080206 TI - Molecule of the Month. Aliskiren fumarate. PMID- 17080207 TI - Perspectives and controversies in the field of stem cell research. AB - The fourth annual meeting of the International Society for Stem Cell Research focused on a number of pressing issues, including: (I) the need to better characterize the biology of stem cells; (II) the need to exploit and optimize the great therapeutic potential of stem cells in tissue regeneration; (III) ethical and safety considerations related to the use of human embryonic stem cells; (IV) the contribution of adult stem cells to carcinogenesis; (V) the need to investigate the biology of cancer stem cells. The purpose of this report is to summarize the current status of stem cell research, as surmised by the proceedings of this meeting. PMID- 17080208 TI - Laboratory methods in the haemostatic laboratory. PMID- 17080209 TI - Monitoring unfractionated heparin with the aPTT: time for a fresh look. AB - Laboratory monitoring is widely recommended to measure the anticoagulant effect of unfractionated heparin and to adjust the dose to maintain levels in the target therapeutic range. The most widely used laboratory assay for monitoring unfractionated heparin therapy is the activated partial thromboplastin time (aPTT). A fixed therapeutic range for the aPTT of 1.5 to 2.5 times the control value has become widely accepted, but the evidence supporting this range is weak and the clinical validity of using the aPTT for predicting thrombotic or bleeding events is questionable. The aPTT test is also affected by numerous preanalytic and analytic variables that are unrelated to the anticoagulant effect of unfractionated heparin, further eroding its potential value for monitoring unfractionated heparin treatment. Unfractionated heparin dose appears to be more important than the aPTT in predicting clinical efficacy. Despite serious limitations, the reliance on the aPTT is likely to continue because of its ready availability and familiarity of clinicians with the test. The focus of clinicians who manage unfractionated heparin therapy should be to ensure that an adequate starting dose of unfractionated heparin is used and that the aPTT method is standardized. Future research efforts should be directed towards developing methods to improve standardization of the aPTT assay for monitoring unfractionated heparin. Direct measures of the concentration of unfractionated heparin in the blood are attractive because these assays are not affected by many of the biologic variables that interfere with the aPTT and may be suitable for automation. However, currently available unfractionated heparin assays are much more expensive than the aPTT, are not widely available, and their validity has not been adequately assessed in clinical outcome studies. PMID- 17080210 TI - Thrombin generation, a function test of the haemostatic-thrombotic system. AB - By the use of a fluorogenic thrombin substrate and continuous calibration of each individual sample, it is now possible to obtain a thrombin generation (TG) curve (or thrombogram) in plasma, with or without platelets, in an easy routine procedure at high throughput and with an acceptable experimental error (<5%). Evidence is growing that the parameters of the thrombogram, and notably the area under the curve (endogenous thrombin potential, ETP), are useful in assessing bleeding- or thrombotic risk and its modification by antithrombotic- or haemostatic treatment. Available data strongly suggest that conditions (congenital, acquired, drug-induced) that increase TG all cause a thrombotic tendency and that conditions that decrease TG prevent thrombosis but, beyond a limit, cause bleeding. Diminution of TG is a common denominator of all antithrombotic treatment, including anti-platelet drugs. The thrombogram can also be used as a tool in the search for new antithrombotics and reflects the haemorrhagic or thrombotic side effects of other drugs (e.g. oral contraceptives). The thrombogram thus is a promising new approach to clinical management of bleeding and thrombotic disease as well as a tool in drug research and epidemiology. Our experience at this moment is insufficient, however, to already clearly define its limits. PMID- 17080211 TI - Use of calibrated automated thrombinography +/- thrombomodulin to recognise the prothrombotic phenotype. AB - There is currently no validated method to detect a prothrombotic phenotype. The question remains, can tissue factor (TF) induced thrombin generation (TG), as measured with the calibrated automated thrombinography (CAT) technique, according to Hemker et al., recognise a prothrombotic state either as such, or when the activated protein C (APC)-system is boosted with thrombomodulin (TM)? We determined the normal range of CAT-TG +/- TM in a group of 71 healthy blood donors, in 11 healthy women using oral contraceptives (OC), and in 89 patients with a history of venous thromboembolism (VTE), divided into a group of 50 in which a prothrombotic risk factor could be found (VTEprf+) and 39 others (VTEprf ). The endogenous thrombin potential (ETP) in the OC, VTEprf+ and VTEprf- group was significantly higher than for the controls. In the presence of TM, the differences were significantly higher than in its absence. The VTEprf+ group had a higher ETP, +/- TM than the VTEprf-group. In conclusion, TG, measured with the CAT technique in the presence of TM is capable of detecting the prothrombotic phenotype with a high sensitivity of 0.93 (95% confidence limits 0.82-0.99). PMID- 17080212 TI - Review: Laboratory markers quantifying prothrombin activation and actions of thrombin in venous and arterial thrombosis do not accurately assess disease severity or the effectiveness of treatment. AB - Thrombin is normally produced for hemostasis and physiological wound healing. Increased thrombin production in vivo, cell activation and inflammation mediated in part by thrombin are hallmarks of both arterial and venous thrombosis. Thrombin generates (pro) coagulant, mitogenic, inflammatory and anticoagulant responses by interacting with a variety of cells in vivo. Both direct and indirect thrombin inhibitors are effective drugs for preventing and treating the consequences of arterial and venous thrombosis. For these reasons, measurements of the production and activities of thrombin in vivo have the potential for gauging the extent of thromboembolism and the responses of patients to anticoagulant, antiplatelet and anti-inflammatory drugs. However, a critical review of published information suggests that measurement of thrombin production and activity in patients at risk for and in patients with significant thrombosis generally does not provide information useful for clinical decision-making. This lack of clinical utility of levels of thrombin production in vivo may arise from two causes: the inability of the measurement to differentiate between physiological (hemostatic) and disease-related (pathological) sources and/or causes of thrombin productio n in vivo, and the inability of antithrombotic treatment modalities to permanently eliminate the stimuli that cause increased thrombin production evident in venous and arterial thrombosis. PMID- 17080213 TI - Evaluation of ProC Global assay in women with a history of venous thromboembolism on hormonal therapy. AB - The risk of thrombosis in women increases significantly during treatment with hormonal therapy (HT). The aim of this study was to evaluate ProC Global assay in women with a history of venous thromboembolism (VTE) while using HT. Protein C activation time normalized ratio (PCAT-NR) levels were significantly lower in 32 women with a history of VTE while using HT (0.72 +/- 0.1) compared with 56 healthy controls without HT, matched by age at blood sampling (0.99 +/- 0.2) and 40 healthy controls with HT, matched by age and HT at VTE event (0.94 +/- 0.2) (P < 0.001 for both). PCAT-NR lower than the cut-off level of 0.8 was found in 23/32 (72%) patients compared with 5/56 (9%) age-matched controls (OR = 26, 95%CI: 7 106, P < 0.001) and 9/40 (22.5%) of HT-matched controls (OR = 9, 95%CI: 2.7-30, P < 0.001). Any thrombophilic risk factor was found in 20/32 (62.5%) of patients compared with 12/56 (21.4%) of agematched controls (OR = 6, 95%CI: 2.1-10, P < 0.001) and 12/40 (30%) of HT-matched controls (OR = 4, 95%CI: 1.3-11.8, P = 0.006). Out of the variables that are risk factors of VTE as age, HT or thrombophilic risk factor, ProC Global assay was found in the multivariate analysis - logistic regression, as the parameter that was the most associated with patient group [Exp(B) = 15.8, 95% CI: 4.2-59.0, P < 0.001]. In conclusion, abnormal PCAT-NR is associated with VTE in women using HT. ProC Global assay may potentially serve as a diagnostic tool for evaluating the risk of VTE in women prior to administration of HT. PMID- 17080214 TI - Performance goals for the laboratory testing of antithrombin, protein C and protein S. AB - To achieve a reliable analytical quality for both monitoring and diagnostic testing, laboratories need to fulfil the widely accepted analytical performance goals based on the biological variation of the analytes of testing. Not only is the short-term analytical performance, which regularly is assessed by internal quality control procedures, of importance, but also the long-term analytical performance. To assess the long-term analytical performance, data obtained from an external quality assessment programme can be used. In this study we have used the evaluation model designed by the ECAT Foundation for the assessment of the longterm analytical performance, including imprecision, bias and total analytical error. The model was applied to the data from 136 different laboratories for the assay of antithrombin (activity), protein C (activity and antigen) and protein S (activity, total and free antigen). The imprecision (median; range), reflected by the long-term analytical coefficient of variation (LCV (A) ), was the lowest for antithrombin (7.6%; 2.6 - 43.8%) and the highest for protein S activity (17.2%; 4.3 - 88.6%). For bias and total error the same pattern was observed (antithrombin: 3.8%; 0.3 - 17.1% and 9.1%; 3.4 - 34.3%, respectively; protein S activity: 12.8%; 3.1 - 34.8% and 24.5%; 9.9 - 87.0%, respectively). For the majority of the laboratories (70 - 85%) the imprecision contributes considerably more to the total error than the bias. However the effect of the bias on the analytical quality is not negligible. Assays for antithrombin, protein C and protein S are mainly used for diagnostic testing. About 70 - 100% of the laboratories can fulfil the desirable performance goal for imprecision. The desirable performance goal for bias was reached by 50 - 95% of the laboratories. In all cases the highest numbers of laboratories fulfilling performance goals was obtained for the protein C variables. To improve the analytical quality in assays of antithrombin, protein C and protein S it is highly recommended that primarily imprecision (non-systematic failures) be suppressed. However the effect of the bias (systematic failures) on the analytical quality should not be neglected. A useful tool for determining the imprecision (LCV (A) ) and bias is the long-term analytical performance evaluation model as used by the ECAT Foundation. PMID- 17080215 TI - Point-of-care International Normalised Ratios: UK NEQAS experience demonstrates necessity for proficiency testing of three different monitors. AB - External quality assessment (EQA) or proficiency testing is widely considered to be necessary for International Normalised Ratio (INR) determinations performed in conventional laboratory settings. There is increasing use of near-patient-test (NPT) or point-of-care (POC) INR devices and it is not known whether EQA is also necessary for these monitors. We report here on six years experience of proficiency testing for POC monitors used by health care professionals. Three devices were used by >10 centres who participated in the programme, the CoaguChek (CUC), the CUC-S and the TAS or Rapidpoint Coag. Not all users of the same type of monitor obtained the same INR result when analysing the same plasma sample. For the three monitors the CV of results in different centres was 11-14%. The variation between results in different centres could relate to inappropriately handled proficiency testing material, inaccuracies in the calibration of the system by the manufacturer or deterioration during transport/storage of the test strips. In each survey 10-11% of centres using POC monitors obtained INR results which were >15% different from those in other centres using the same monitors. For hospital laboratories using conventional INR techniques this figure was 12%. The relationship between INR results obtained by users of the Rapidpoint Coag or TAS monitor and results obtained by conventional techniques was not constant over the period of study. During one period INRs with TAS were 13.7% greater than with conventional methods. For the remaining three time periods results were similar. Our data suggest that the variation between INR results determined with three POC monitors show similar variation to that observed in hospital laboratories using conventional methods. Based on our data we recommend that users of these POC monitors participate regularly in an independent external proficiency testing programme. PMID- 17080216 TI - The UK National External Quality Assessment Scheme (UK NEQAS) for molecular genetic testing in haemophilia. AB - Molecular genetic analysis of families with haemophilia and other inherited bleeding disorders is now a common laboratory investigation. In contrast to phenotypic testing in which strict quality control is adhered to, in haemophilia molecular genetic testing there has been a lack of any external quality assurance schemes. In 1998 the UK National External Quality Assessment Scheme (UK NEQAS) established a pilot quality assurance scheme for molecular genetic testing in haemophilia. Results from three initial surveys highlighted problems with the quality of samples when used to screen for the intron 22 inversion within the F8 gene. The scheme was re-launched in 2003, and since that time there have been five exercises involving whole blood or immortalised cell line DNA. The results together with an overall summary of the exercise are subsequently returned to participants. Exercises to date have focused exclusively on haemophilia A and QA, material has included screening for the intron 1 and intron 22 inversions as well as sequence analysis. A paper exercise circulated in 2003 highlighted problems with the format of reports and, following feedback to participants, only a single error has been made in the subsequent four exercises. Participating laboratories now receive QA material every six months. Immortalised cell line material was introduced in 2005 and was shown to perform well. This will allow expansion of the scheme and a reduction in the dependence on blood donation. PMID- 17080217 TI - Rat models of myocardial infarction. Pathogenetic insights and clinical relevance. AB - Animal models of cardiovascular pathology contribute towards understanding and treatment of a broad range of conditions. Specifically in the context of acute myocardial infarction (AMI), rat models have been commonly used in studies of pathogenesis, investigation and novel therapies, although there has often been difficulty in translating experimental findings to clinical benefit. However, recent years have seen two important changes to our clinical approaches to AMI. First, there is increasing recognition that the pathophysiology of human AMI is a process occurring at many levels, not just within the epicardial coronary artery, but also within the microvasculature and the myocardium. Second, contemporary treatments are shifting away from thrombolytic dissolution of epicardial coronary thrombus to direct mechanical approaches using angioplasty and stents. These changes in our understanding of AMI have implications for the relevance of these animal models. The following discussion therefore reviews and examines the current rat models of AMI, places them in a clinical context, discusses their advantages and limitations, and outlines likely future developments, providing an overview of the place of these important models of AMI. PMID- 17080218 TI - Management of cutaneous type IV hypersensitivity reactions induced by heparin. AB - Localized hypersensitivity reactions to subcutaneous heparin injections have been described since 1952. Yet, the incidence of these reactions, which are distinct from skin lesions associated with heparin-induced thrombocytopenia type II (HIT II), remains uncertain. However, in the last 10 years an increasing number of patients have been reported, leading to the assumption that cutaneous hypersensitivity reactions towards heparin are underreported. Clinically patients present with itching, sometimes infiltrated, and blistering erythemas at the injection sites of heparins. The diagnosis of cutaneous heparin allergy may, on the one hand, lead to delay of required medical or surgical treatment. On the other hand, delayed initiation of treatment may lead to a generalized eczematous reaction. Hence, from review of 223 cases of patients with cutaneous hypersensitivity reactions to heparin, we here summarize the clinical picture of cutaneous type IV allergic reactions, define risk factors on both the patient- and drug-side, and give an overview of principle therapeutic alternatives, as well as recommendations for treatment options for emergency and elective patients. As the proposed management of patients with cutaneous hypersensitivity reactions to heparin may have fatal consequences when applied in patients with HIT type II, diagnosis of skin lesions in heparin-treated patients needs to be precise. PMID- 17080219 TI - Immunohistochemistry of thrombi following iliac venous stenting: a novel model of venous thrombosis. AB - Stenting has become a common intervention for venous occlusive disease. Little is known regarding the composition of venous thrombi complicating stent placement. The optimal design of antithrombotic agents in this setting requires this knowledge. Quantitative immunohistochemistry was undertaken to define the platelet, fibrin(ogen) and leukocyte composition and spatial orientation of venous thrombi following percutaneous iliac stent placement in pigs. Venous stent thrombus size was measured by weight and scintillation detection of autologous (111) In-platelets. Thrombi were divided in segments (cephalad to caudad), sectioned and stained with monoclonal anti-platelet glycoprotein Ib or polyclonal anti-fibrin(ogen) fluorescent antibodies. Thrombus platelet content was 100-fold greater than paired whole blood samples. The caudal-most segments contained platelet-rich aggregates (p < 0.05) with abundant leukocytes (p < 0.0001) relative to more cephalad segments. Platelet and fibrin(ogen) content varied over an eight-fold range between segments but were directly correlated with each other (r = 0.77; p < 0.0001). The platelet co-localization with fibrin(ogen) is consistent with the phospholipid dependence of prothrombin activation. The abundance and caudal distribution of platelet-leukocyte aggregates indicate their preferential accretion from flowing blood early in the genesis of venous stent thrombi. These may represent novel cellular targets for the prevention and treatment of venous thrombosis. PMID- 17080220 TI - The effect of glycaemic control on fibrin network structure of type 2 diabetic subjects. AB - Diabetic subjects have been shown to have altered fibrin network structures. One possible cause may be fibrinogen glycation resulting in altered structure/function properties. We investigated the effect of glucose control on fibrinogen glycation and fibrin network structure in type 2 diabetes. Blood samples were taken from twenty uncontrolled diabetic subjects at baseline to determine the levels of fibrinogen glycation and fibrin network structures. The subjects were then treated with insulin until blood glucose control was achieved before end blood samples were taken. Twenty age- and BMI-matched non-diabetic subjects were included as a reference group. The diabetic subjects had significantly higher mean fibrinogen glycation at baseline than the non-diabetic subjects (7.84 vs. 3.89 mol glucose / mol fibrinogen; p < 0.001). This was significantly reduced during the intervention (7.84 to 5.24 mol glucose / mol fibrinogen; p < 0.0002) in the diabetic group. Both groups had high mean fibrinogen concentrations (4.25 and 4.02 g/l, diabetic and non-diabetic subjects respectively). There was no difference in fibrinogen concentration, porosity, compaction and kinetics of clot formation between the diabetic subjects and non diabetic subjects at baseline, nor were there any changes during the intervention despite the reduced fibrinogen glycation. Fibrin network characteristics correlated well with fibrinogen but not with any markers of glycaemic control. Improved glycaemic control resulted in decreased fibrinogen glycation but not fibrinogen concentration. It seems as though porosity, compaction and kinetics of clot formation are more related to fibrinogen concentration than fibrinogen glycation in this model. PMID- 17080221 TI - An investigation of the von Willebrand factor genotype in UK patients diagnosed to have type 1 von Willebrand disease. AB - Forty families diagnosed by UK centres to have type 1 VWD were recruited. Following review, six families were re-diagnosed to have type 2 VWD, one to have a platelet storage pool disorder, and one family was determined to be unaffected. Direct DNA sequencing of the promoter region and all exons and intronic boundaries of the VWF gene identified six mutations likely to be causative of VWD in index cases of nine of the 32 (28%) confirmed type 1 VWD families. These included R1205H (3614G > A) VWD Vicenza, P1648fsX45 (4944delT), D141G (422A > G) and three splice site mutations: 3108 + 5G > A, 7437 + 1G > A and 3379 + 1G > A. The Y1584C (4751A > G) polymorphism was present in eight additional families. No significant VWF gene mutation or polymorphism was identified in 15 of the 32 type 1VWD index cases (47%). Haplotype studies were performed using a panel of VWF polymorphisms to investigate the segregation in families of VWD phenotype with the VWF gene. In 13 of the 32 families it was likely that VWD segregated with the VWF gene. In eight families (25%) VWD clearly did not segregate with the VWF gene. We suggest that mutation screening of the VWF gene has limited general utility in genetic diagnostic and family studies in type 1 VWD. If genetic studies are performed, the incomplete penetrance and variable expressivity of type 1 VWD must be taken into account. Unless linkage of VWD phenotype with the VWF gene can be clearly demonstrated, the results of any genetic family studies should be interpreted with caution. PMID- 17080222 TI - Integrin binding characteristics of the disintegrin-like domain of ADAM-15. AB - Although discovered as potent inhibitors of a (IIb) ss (3) -mediated platelet aggregation, snake venom disintegrins are now known to bind to other integrins according to different degrees of potency and specificity. More recently, homologues of the disintegrinlike loop have been found as a discrete domain in the ADAM family, yet the potency and specificity of each of these domains in terms of integrin binding is relatively unknown. In this present study, we have selected the disintegrin-like domain (dd) of ADAM-15 (designated as ddADAM-15), the only RGD containing domain in the ADAM family, for a structure/function study. Experimentally, the ddADAM-15 and a number of mutants in which the RGD containing loop was substituted by cognate regions from ADAM-2, -12 and -19 were tested in terms of integrin-binding activity. For comparison with ADAM-15, an additional mutant (dd (den) ADAM-15) was designed based upon the RGD-containing loop of snake venom dendroaspin, a disintegrin-like integrin antagonist. The results showed that ddADAM-15 is an inhibitor of platelet aggregation, though with less potency than dd (den) ADAM-15. None of the other mutants exhibited significant inhibition of platelet aggregation. ddADAM-15 was found to have higher binding ability for a (2) ss (1) and a (9) ss (1) than the ADAM-2 derived mutant which appeared to be more selective for a (V) ss (3) and a (4) ss (1) than either ddADAM-15 or its ADAM-19 based mutant. The integrin-binding properties of ddADAM-15 were completely abolished by point mutation within the RGD motif (R (64) GD ? A (64) GD). These results suggest a more subtle contribution of this loop sequence to defining the functionality of the ADAMs compared to dendroaspin. PMID- 17080223 TI - Platelet-induced growth of human fibroblasts is associated with an increased expression of 5-lipoxygenase. AB - Proliferation of fibroblasts is vital for adequate wound healing but is probably also involved in different hyperproliferative disorders such as atherosclerosis and cancer. The regeneration of tissue usually starts with coagulation, involving release of mitogenic and inflammatory factors from activated platelets. This study focuses on the role of eicosanoids in the proliferative effects of platelets on human fibroblasts. We show that the phospholipase A (2) inhibitor 7,7-dimethyl-5,8-eicosadienoic acid (DMDA), the combined cyclooxygenase (COX) and lipoxygenase (LOX) inhibitor 5,8,11,14-eicosatetraynoic acid (ETYA) and the LOX inhibitor 5,8,11-eicosatriynoic acid (ETI) block the platelet-induced proliferation of serum starved subconfluent human fibroblasts. Anti-proliferative effects were also obtained by specific inhibition of 5-LOX with 5,6-dehydro arachidonic acid (5,6-dAA), whereas the 12-LOX inhibitor cinnamyl-3,4-dihydroxy- a -cyanocinnamate (CDC) did not affect the platelet-stimulated growth of fibroblasts. The expression of 5-LOX was analyzed by reverse-transcriptase mediated PCR (RT-PCR), Western blotting and HPLC. 5-LOX message and protein was detected in fibroblasts but not in platelets. Incubation with platelets markedly increased, already after one hour, the expression of 5-LOX in the fibroblast culture. The increased 5-LOX activity was associated with an elevated level of the 5-LOX metabolite 5-hydroxyeicosatetraenoic acid (5-HETE) reaching its maximum after 1 - 2 hours of co-incubation of fibroblasts and platelets. The 5-HETE production was reduced by the inhibitors DMDA, ETYA and ETI. In conclusion, this study suggests that platelet-stimulated proliferation of fibroblasts is mediated by an increased 5-LOX activity, which supports recent findings indicating a crucial role for this enzyme in proliferative disorders such as atherosclerosis. PMID- 17080224 TI - No difference in the effects of clopidogrel and aspirin on inflammatory markers in patients with coronary heart disease. AB - Aspirin reduces several pro-inflammatory markers in patients with coronary heart disease (CHD), while limited data exists with clopidogrel. The aim of the present substudy of ASCET was to assess the influence of clopidogrel as compared to aspirin on selected circulating inflammatory markers in patients with stable angiographically verified CHD. Patients on treatment with aspirin 160 mg/day for at least seven days were randomized to either aspirin 160 mg/day (n = 105) or clopidogrel 75 mg/day (n = 101). Fasting blood samples were drawn at baseline and after one month and after one year for determination of high sensitivity C reactive protein, tumor necrosis factor a (TNFa), interleukin 6, monocyte chemoattractant protein 1(MCP-1), CD40L, P-selectin, interleukin 10 and transforming growth factor. The groups were similar regarding demographic variables. There were no differences in any variables including changes from baseline to one month and one year between the groups. In the aspirin group we found significantly lower levels of TNFa and MCP-1 after one year; 1.00 versus 1.16 pg/ml (p < 0.001) and 245 versus 261 pg/ml (p < 0.001), respectively. Likewise, in the clopidogrel group the level of TNFa was significantly reduced after one year; 0.99 versus 1.19 pg/ml (p < 0.001). In patients with CHD we found no between-group differences in circulating markers of inflammation after one year treatment with clopidogrel 75 mg/day compared to aspirin 160 mg/day, but in both groups lower levels of TNFa were obtained. The present results indicate similar anti-inflammatory effects of the two drugs. PMID- 17080225 TI - Plasminogen activator inhibitor-1 impairs plasminogen activation-mediated vascular smooth muscle cell apoptosis. AB - The role of plasminogen activator inhibitor-1 (PAI-1) in vascular smooth muscle cell (VSMC) apoptosis mediated by plasminogen activation was studied with the use of aorticVSMC derived from mice with deficiency of PAI-1 (PAI-1 (-/-) ), tissue type (t-PA (-/-) ) or urokinase-type (u-PA (-/-) ) plasminogen activator or from wildtype (WT) mice with corresponding genetic background. Plasminogen incubated with confluent VSMC was activated in a concentration-dependent and saturable manner for all four cell types, with maximal activation rates that were comparable for WT, u-PA (-/-) and t-PA (-/-) cells, but about two-fold higher for PAI-1 (-/-) cells. Plasminogen activation was impaired by addition of the lysine analogue 6-aminohexanoic acid, and by addition of t-PA and u-PA neutralizing antibodies, suggesting that it depends on binding to cell surface COOH-terminal lysine residues, and on plasminogen activator activity. Morphological alterations consistent with apoptosis were observed much earlier in PAI-1 (-/-) than in WT VSMC. Without addition of plasminogen, the apoptotic index was similar for all four cell types, whereas after incubation with physiological plasminogen concentrations, it was greater in PAI-1 (-/-) VSMC, as compared to WT, t-PA (-/-) or u-PA (-/-) VSMC. Furthermore, the apoptotic rate paralleled the release of plasmin. Thus, plasmin-mediated apoptosis of VSMC occurs via plasminogen activation by either t-PA or u-PA and is impaired by PAI-1. PMID- 17080226 TI - Rational humanization of the powerful antithrombotic anti-GPIbalpha antibody: 6B4. AB - Fab-fragments of the monoclonal antibody 6B4, raised against human glycoprotein Ibalpha (GPIbalpha), have a powerful antithrombotic effect in baboons by blocking the GPIbalpha binding site for von Willebrand factor (VWF), without significant prolongation of the skin bleeding time. In order to bring this antibody to the clinic,we here humanized for the first time an anti-human GPIbalpha by variable domain resurfacing guided by computer modeling. First, the genes coding for the variable regions of the heavy and light chains of 6B4 were cloned and sequenced. Based on this, a three-dimensional structure of the Fv-fragment was constructed by using homology-based modeling, and with this and comparison with antibodies with known structure,"murine" putative immunogenic residues which are exposed, were changed for "human-like" residues. The humanized Fab-fragment, h6B4-Fab, was constructed in the pKaneo vector system, expressed and purified and showed in vitro an unaltered, even slightly higher binding affinity for its antigen than the murine form as determined by different ELISA set-ups and surface plasmon resonance. Finally, injection of doses of 0.1 to 1.5 mg/kg of h6B4-Fab in baboons showed that both pharmacokinetics and ex-vivo bio-activity of the molecule were to a large extent preserved. In conclusion,the method used here to humanize 6B4 by resurfacing resulted in a fully active derivative, which is now ready for further development. PMID- 17080227 TI - Fibrinogen Darlinghurst: hypofibrinogenaemia caused by a W253G mutation in the gamma chain in a patient with both bleeding and thrombotic complications. PMID- 17080228 TI - Discrepancy between capillary and venous International Normalised Ratio (INR) in a patient on vitamin-K antagonist: potentially serious clinical consequences. PMID- 17080229 TI - Pulmonary embolism associated with olanzapine. PMID- 17080230 TI - Characterization of an acquired factor VIII inhibitor and plasmapheresis therapy in a patient with bullous pemphigoid. PMID- 17080231 TI - Extrinsic venous compression: a sufficient explanation for venous thromboembolism due to massive fibroids? PMID- 17080232 TI - Increased incidence of renal cysts in patients with abdominal aortic aneurysms: a common pathogenesis? AB - We have empirically observed that patients with abdominal aortic aneurysms (AAAs) seem to have an increased incidence of renal cysts on computed tomography (CT). In order to evaluate this possible association, a retrospective cohort study was conducted comparing the incidence of renal cysts on CT scan in 100 patients with AAA to 100 patients without AAA (matched by age and gender). Univariate analysis and multiple logistic regression were performed to evaluate the association of AAAs and other risk factors with the presence of renal cysts. Of patients with AAAs, 54% had renal cysts compared to only 30% in the control group (p = 0.0006, relative risk = 2.73). The AAA group had a higher incidence of chronic obstructive pulmonary disease (COPD, 14% vs. 1%), hypertension (76.6% vs. 46.5%), coronary artery disease (38.3% vs. 12%), and hypercholesterolemia (41.5% vs. 9.1%) compared to the non-AAA group. There was a significant linear correlation between renal cysts and COPD (p = 0.011), the presence of AAA (p = 0.0005), and age (p = 0.019), whereas hypercholesterolemia (p = 0.059) and diabetes (p = 0.063) approached significance. On multivariate analysis, there were three independent predictors of renal cysts: COPD (p = 0.051), age (p = 0.01), and AAA (p = 0.028). In conclusion, there is a significantly higher incidence of renal cysts in patients with AAA compared to patients without AAA. To our knowledge, this association has not previously been reported. Future studies are needed to determine whether this correlation is the result of a commonality in the pathogenesis of AAA and renal cysts. PMID- 17080233 TI - A ruptured pancreaticoduodenal artery aneurysm repaired by combined endovascular and open techniques. AB - Aneurysms of the pancreaticoduodenal arteries (PDA) are rare, accounting for <2% of all visceral aneurysms. An association with celiac artery stenosis has been reported. Many present with rupture, and a high mortality can be expected. Treatment is therefore challenging. Arterial ligation, anuerysmectomy, or bypass has been the mainstay of treatment. We recently treated a patient (who had no celiac axis) with a ruptured PDA aneurysm with combined open and endovascular techniques. A 46-year-old man was transferred to our hospital with a 1-day history of abdominal pain and syncope. On admission, an abdominal and pelvis computerized tomographic (CT) scan identified a large mesenteric hematoma, a 1.9 cm PDA aneurysm, and an occluded celiac axis. Mesenteric angiography revealed no active aneurysm leak and a stenotic superior mesenteric artery (SMA) origin. All hepatic blood flow originated from the stenotic SMA via markedly enlarged PDA collaterals. The patient was brought to the operating room, where absence of the celiac axis was confirmed. An aorto-to-proper hepatic and SMA bypass was performed using a bifurcated polyester graft. The next day, the patient was brought to the angiography suite, where the PDA aneurysm was coiled. Postprocedure CT scans confirmed thrombosis of the aneurysm. Ruptured mesenteric artery aneurysms are a challenging problem for the vascular surgeon. PDA aneurysms are rare and often occur in an unfavorable location. There appears to be an association with anatomic anomalies of the mesenteric circulation. Prompt invasive and noninvasive diagnostic studies aid in the definitive management of this often fatal problem. Combined endovascular and open techniques can be used for successful treatment. PMID- 17080234 TI - Outcome of metastatic GIST in the era before tyrosine kinase inhibitors. AB - BACKGROUND: Treatment of metastatic GIST with imatinib mesylate results in a 2 year survival of approximately 72%. The outcome of patients with metastatic GIST not treated with tyrosine kinase inhibitors is not well defined. METHODS: One hundred nineteen patients with metastatic GIST diagnosed prior to July 1, 1998 (approximately 2 years prior to the use of imatinib for GIST) were identified from an institutional database of patients with pathologically confirmed GIST. Mutational analysis was performed in cases with available tissue. The log rank test and Cox regression models were used to assess prognostic factors. RESULTS: Median survival was 19 months with a 41% 2-year survival and a 25% 5-year survival. Resection of metastatic GIST was performed in 81 patients (68%), while 50 (42%) received conventional chemotherapy. Twelve patients (10%) were eventually started on imatinib. Primary tumor size <10 cm, <5 mitoses/50 HPF in the primary tumor, epithelioid morphology, longer disease-free interval, and surgical resection were independent predictors of improved survival on multivariate analysis. Mutational status did not predict outcome. In patients who underwent resection, the 2 year survival was 53%, and negative microscopic margins also independently predicted improved survival. CONCLUSIONS: Treatment with imatinib appears to improve 2-year survival of metastatic GIST by approximately 20% when compared to surgery alone. The combination of imatinib and surgery for the treatment of metastatic GIST therefore warrants investigation. PMID- 17080235 TI - Disparities in colon cancer presentation and in-hospital mortality in Maryland: a ten-year review. AB - BACKGROUND: Much attention has focused on in-hospital treatment disparities in colon cancer outcomes. Little is known about the effect of prehospital factors on outcomes. We hypothesized that racial and socioeconomic disparities exist in the presentation of colon cancer and that these disparities affect in-hospital outcomes. METHODS: Ten-year data on colon cancer patients were obtained from the Maryland Hospital Discharge Database. Life-threatening symptoms at presentation served as a proxy for delay in diagnosis. Patients with the primary diagnosis of colon cancer treated with surgical resection were included. Outcomes of interest were obstruction, hemorrhage, perforation, and in-hospital mortality. RESULTS: A total of 14,291 patients had primary colon cancer, and 13,031 underwent resection. Among this group, 52% were male, 22% were African American (AA), and mean age of AA was 66.0 years versus non-AA mean age of 70.5 years (P < .001). Overall, 27.6% of patients presented with life-threatening symptoms. In-hospital mortality rate was 3.8%. Symptomatic patients had a 2-fold higher rate of in hospital mortality (odds ratio [OR], 6.06 vs. 2.89, P < .001). Multivariate analysis demonstrated that AA were more likely to have life-threatening symptoms at presentation independent of socioeconomic status (SES) (OR, 1.36). In addition, AA had a higher in-hospital mortality, both overall (OR, 1.39) and in the higher SES (OR, 1.81). CONCLUSIONS: Racial disparities exist in the rate of presentation with life-threatening symptoms that may be a proxy for a delay in diagnosis. These findings were independent of SES, implying that factors beyond health care access may account for poorer outcomes among AA. PMID- 17080236 TI - Stromal osteonectin overexpression is associated with poor outcome in patients with ampullary cancer. AB - BACKGROUND: Osteonectin has been suggested to be important in the progression of pancreatic cancer but has not been correlated with survival. We determined the osteonectin expression and its influence on survival in patients with ampullary carcinoma. METHODS: Tissue microarrays were constructed from the tumors of 56 patients with ampullary cancer undergoing pancreaticoduodenectomy. Immunohistochemical staining for osteonectin was undertaken and compared with staining in chronic pancreatitis (n = 13) and normal pancreas (n = 19). Survival curves were created by the Kaplan-Meier method and compared by log rank analysis. Median follow-up for all living patients with ampullary cancer was 69.6 months. RESULTS: Osteonectin was significantly (P < .05, Fisher's exact test) overexpressed in the stroma of ampullary cancers (90%) relative to chronic pancreatitis (62%) and normal pancreas (0%). Tumors expressing osteonectin were more likely to have nodal metastases than those lacking osteonectin expression (48% vs. 0%, P = .06, Fisher's exact test) and showed decreased survival. Node negative status, pylorus preservation at the time of pancreaticoduodenectomy, and lack of osteonectin expression were predictors of prolonged survival by multivariate analysis. CONCLUSIONS: Although the importance of tumor-stroma interactions in periampullary cancers is not fully understood, our data suggest that osteonectin is an integral stromal element in ampullary cancers, and its overexpression is associated with decreased survival. PMID- 17080237 TI - Radiation-induced cellular DNA damage repair response enhances viral gene therapy efficacy in the treatment of malignant pleural mesothelioma. AB - BACKGROUND: Malignant pleural mesothelioma (MPM) treated with radiotherapy (RT) has incomplete responses as a result of radiation-induced tumoral stress response that repairs DNA damage. Such stress response is beneficial for oncolytic viral therapy. We hypothesized that a combination of RT and NV1066, an oncolytic herpes virus, might exert an additive or synergistic effect in the treatment of MPM. METHODS: JMN, a MPM cell line, was infected with NV1066 at multiplicities of infection of .05 to .25 in vitro with and without radiation (1 to 5 Gy). Virus replication was determined by plaque assay, cell kill by lactate dehydrogenase assay, and GADD34 (growth arrest and DNA damage repair 34, a DNA damage-repair protein) by real-time reverse transcriptase-polymerase chain reaction and Western blot test. Synergistic cytotoxicity dependence on GADD34 upregulation was confirmed by GADD34 small inhibitory RNA (siRNA). RESULTS: Synergism was demonstrated between RT and NV1066 across a wide range of doses. As a result of such synergism, a dose-reduction for each agent (up to 5500-fold) can be accomplished over a wide range of therapeutic-effect levels without sacrificing tumor cell kill. This effect is correlated with increased GADD34 expression and inhibited by transfection of siRNA directed against GADD34. CONCLUSIONS: RT can be combined with oncolytic herpes simplex virus therapy in the treatment of malignant pleural mesothelioma to achieve synergistic efficacy while minimizing dosage and toxicity. PMID- 17080238 TI - Major liver resections synchronous with colorectal surgery. AB - BACKGROUND: Surgical strategy in liver metastases synchronous to colorectal cancer remains controversial. The aim of this study was to evaluate feasibility and short-term outcomes of major hepatectomies synchronous to colorectal surgery. METHODS: Between January 1985 and December 2004, 79 patients underwent major hepatectomy for metastases synchronous to colorectal cancer; 31 underwent synchronous hepatectomy and colorectal surgery, and 48 underwent delayed liver resection. RESULTS: The synchronous group had a higher rate of right colectomy (38.7% vs. 18.8%, P = .0499) and larger metastases (8 vs. 5.3 cm, P = .0032). Mortality (one patient in synchronous group), morbidity, and anastomotic leak rates were similar in the two groups. Colon-related morbidity did not cause adjunctive liver complications. Hospitalization in delayed hepatectomies was shorter (10.4 days vs. 13.9 days, P = .0021). Blood and plasma transfusions were higher in synchronous resections (41.9% vs. 16.7%, P = .0131 and 54.8% vs. 31.3%, P = .0370); no differences were found in the last 10 years. Considering both surgical procedures (colorectal + liver resection), in delayed hepatectomies, morbidity was higher (56.3% vs. 32.6%, P = .0369) and hospitalization was longer (20.5 vs. 13.9 days, P = .00001). Nine patients underwent major hepatectomy at the same time as anterior rectal resection with no mortality (morbidity 22.2%, mean hospitalization 12.4 days). CONCLUSIONS: Major hepatectomies can be safely performed at the same time as colorectal surgery in selected patients with synchronous metastases with similar short-term results, even in the presence of rectal cancer. PMID- 17080239 TI - Impact of regional lymph node evaluation in staging patients with periampullary tumors. AB - BACKGROUND: Two distinct lymph nodes reproducibly assessed by computed tomography for the evaluation of periampullary tumors are the common bile duct (CBD) node and the gastroduodenal artery (GDA) node. We examined whether radiographical enlargement of either lymph node predicts tumor resectability, nodal metastasis, or patient survival. METHODS: Ninety-four consecutive patients underwent attempted curative resection of periampullary tumors between September 2001 and June 2003. A single radiologist recorded in a retrospective, blinded fashion the short- and long-axis measurements of the CBD and GDA nodes. RESULTS: Sixty-one percent (n = 57) of tumors were resectable by pancreaticoduodenectomy. Overall, actual 6-, 12-, and 18-month survival was 87%, 68%, and 63%, respectively. Enlarged radiographical nodal size by either axis was not associated with the presence of metastasis to these lymph nodes or with reduced overall patient survival. Only a CBD node short-axis size >10 mm predicted unresectability (odds ratio, 3.2; P = .036). Liver metastasis and/or carcinomatosis were present in 43% of unresectable patients, and this was associated with decreased survival at both 1 year (25% vs. 77%; P < .001) and 18 months (19% vs. 72%; P <.001). A pathologic diagnosis of metastasis to the GDA node, but not the CBD node, was associated with a similarly decreased survival (1 year: 33% vs. 78%, P = .028; 18 months: 22% vs. 70%, P = .023). CONCLUSIONS: For presumed periampullary malignancy, a CBD node short-axis size >10 mm predicts tumor unresectability. Metastatic disease to the GDA node, particularly for pancreatic adenocarcinoma, portends a poor prognosis equivalent to that of hepatic or peritoneal spread. Given these findings, radiographical CBD lymph node measurements may guide selection for performing laparoscopic staging with or without ultrasonography in conjunction with GDA nodal biopsy in patients with periampullary malignancy. PMID- 17080240 TI - Prognostic value of cytokeratin-positive bone marrow cells of gastric cancer patients. AB - BACKGROUND: Epithelial cells in the bone marrow of patients with gastric cancer suggest tumor dissemination; however, their prognostic implications are controversial. We prospectively evaluated the correlation of bone marrow findings, recurrence rate, and disease-free survival after long-term follow-up. METHODS: Bone marrow were aspirated from both iliac crests and stained with monoclonal cytokeratin (CK)-18 antibody in 209 patients before their initial operation. Patients were followed up for a median of 56 months. RESULTS: Overall, 39 (19%) of 209 patients and 15 (14%) of 109 R0-resected patients had CK-positive cells. CK-positive patients had more local, regional, and distant recurrence than CK-negative patients (P < .05). We found a significantly shorter disease-free survival (P < .05) in the patients with >2 CK-positive cells per 2 x 10(6) bone marrow cells (mean, 35 months) than in patients with 2 CK-positive cells per 2 x 10(6) bone marrow cells was an independent prognostic factor for tumor-related death (P < .05). CONCLUSIONS: Not only the mere presence of CK positive epithelial cells in bone marrow, but also the cell number, correlates with prognosis. Our findings suggest that classifying CK-positive bone marrow cells in these patients will facilitate future studies. PMID- 17080241 TI - Review: Genetically modified plants for the promotion of human health. AB - Plants are attractive biological resources because of their ability to produce a huge variety of chemical compounds, and the familiarity of production in even the most rural settings. Genetic engineering gives plants additional characteristics and value for cultivation and post-harvest. Genetically modified (GM) plants of the "first generation" were conferred with traits beneficial to producers, whereas GM plants in subsequent "generations" are intended to provide beneficial traits for consumers. Golden Rice is a promising example of a GM plant in the second generation, and has overcome a number of obstacles for practical use. Furthermore, consumer-acceptable plants with health-promoting properties that are genetically modified using native genes are being developed. The emerging technology of metabolomics will also support the commercial realization of GM plants by providing comprehensive analyzes of plant biochemical components. PMID- 17080242 TI - Correlation of GLUT-1 overexpression, tumor size, and depth of invasion with 18F 2-fluoro-2-deoxy-D-glucose uptake by positron emission tomography in colorectal cancer. AB - We investigated the wide variability of 18F-2-fluoro-2-deoxy-D: -glucose (FDG) uptake, semiquantified as standardized uptake value (SUV), in positron emission tomography (PET) scanning, in 20 patients with colorectal cancer (CRC), including 1 with synchronous hepatic metastasis. The sensitivity of PET in CRC diagnosis was 100%, with a mean SUV of 8.0 (3.1-11.9). Tumor size and depth of invasion were associated with higher SUVs (P=.0004, .042, respectively). Strong glucose transporter-1 (GLUT-1) expression had significantly positive correlation with the SUV (r=.619, P=.003). GLUT-1 expression revealed positive staining in 17 (85%) of the 20 primary lesions. The central part of the tumor, thought to be relatively hypoxic, had stronger GLUT-1 expression and a higher SUV than the periphery, in both the primary tumor and hepatic metastatic foci. Our data suggest that the SUVs of FDG uptake in PET may be a noninvasive biomarker for advanced CRC, indicative of a large hypoxic tumor with deep invasion. PMID- 17080244 TI - The relationship between intestinal hypoperfusion and serum d-lactate levels during experimental intra-abdominal hypertension. AB - Intra-abdominal hypertension (IAH) and abdominal compartment syndrome (ACS) may result from several clinic situations and carries high morbidity and mortality risk, particularly in intensive care unit patients. The clinical spectrum changes from splanchnic hypoperfusion and intestinal ischemia to multiple organ failure. Previous studies demonstrated that serum D-lactate levels may be an early indicator in intestinal ischemia. This study aimed to investigate the relationship between intestinal ischemia and serum D-lactate levels during experimental IAH. Thirty-two male Wistar Albino rats weighing 250+/-50 g were divided into four groups. Three different intra-abdominal pressure (IAP) levels supplied by placement of an intraperitoneal Peritofix catheter and iso-osmotic polyethylene glycol infusion. Each of the IAP levels (15, 20, and 25 mm Hg groups) was checked with the monitor system and fixed for an hour. Control-group animals were not subjected to increased IAP. One hour later, 5-ml blood samples were taken for measurement of serum D-lactate levels and 2-cm intestinal tissue samples were taken 5 cm proximal to the ileocecal valve for histopathologic examination. Elevated serum D-lactate levels were recorded in animals with higher IAP levels. There was a positive correlation between serum D-lactate levels and IAP levels. Histological examinations of the intestinal tissue samples showed no significant pathologic changes in concordance with intestinal ischemia. Serum D lactate levels may be an early indicator for increased IAP pressure before intestinal ischemic changes occur. PMID- 17080243 TI - The restorative effect of taurine on experimental nonalcoholic steatohepatitis. AB - Our objective was to explore the restorative effect of taurine on experimental nonalcoholic steatohepatitis (NASH). Thirty-six SD rats were randomly divided into three groups, 12 in each group: the normal group was fed standard rat diet; the model group and the treatment group were both fed a high-fat rat diet for 12 weeks, and the rats in the treatment group were simultaneously injected with taurine subcutaneously for 8 weeks. Hepatic histological change was observed; TNF alpha and TGF-beta(1) protein expression was identified by immunohistochemistry; mRNA expression of TNF-alpha, TGF-beta(1), type I procollagen, and adiponectin was measured by RT-PCR; body weight, weight gain, liver weight, and liver index were measured; and biochemical parameters monitored included serum transaminases, serum lipids, fasting plasma glucose, and hepatic level of oxidative stress. Rats in the model group showed a significant increase in liver weight, liver index, serum transaminase activities, serum triglyceride, fasting plasma glucose, and oxidative stress; the mRNA expression of TNF-alpha, TGF-beta(1), and type I procollagen increased, whereas the expression of adiponectin decreased significantly, compared with that in the normal group. The typical hepatic lesions of NASH were observed histologically in the model group. Taurine treatment resulted in a significant decrease in liver weight, liver index, serum transaminase activities, serum triglyceride, fasting plasma glucose, and oxidative stress; the mRNA expression of TNF-alpha, TGF-beta(1), and type I procollagen decreased, but the expression of adiponectin increased significantly, compared with that in the model group. Histological improvement was observed in the treatment group. In conclusion, taurine could inhibit lipid peroxidation, improve lipid and glucose metabolism, decrease synthesis of TNF-alpha and TGF beta(1), promote synthesis of adiponectin, and have a restorative effect on experimental NASH. PMID- 17080245 TI - Hepatic venous pressure gradient in cirrhosis: correlation with the size of varices, bleeding, ascites, and child's status. AB - The hepatic venous pressure gradient (HVPG) clearly reflects portal pressure in cirrhotic portal hypertension. Its relation with variceal bleeding has been well studied. We undertook to study the relation of HVPG to variceal size, Child's status, and etiology of cirrhosis. Patients with cirrhotic portal hypertension with esophageal varices underwent HVPG measurement as part of a prospective evaluation. One hundred seventy-six cirrhotics with varices (M:F, 140:36; mean age, 42.6 +/- 13.4 years), 104 with CLD related to viral etiology, 40 with alcoholic liver disease, 26 cryptogenic with cirrhosis, and 6 with miscellaneous causes of CLD underwent HVPG measurement. The mean HVPG was lower in patients with small varices (n = 77; 14.6 +/- 5.9 mm Hg) than in patients with large varices (n = 99; 19.2 +/- 6.6 mm Hg; P < 0.01). In patients with large varices, the mean HVPG in bleeders (n = 37) was higher than in nonbleeders (n = 62) (21.7 +/- 7.2 vs 17.9 +/- 6.2 mm Hg; P < 0.01). The mean HVPG was significantly higher in Child's B (n = 97; 17.4 +/- 6.9 mm Hg) and C (n = 56; 19.0 +/- 5.7 mm Hg) compared to Child's A cirrhotics (n = 23; 12.2 +/- 5.9 mm Hg; P < 0.01), and Child's C compared to Child's B cirrhotics (P = 0.05). HVPG was higher in alcoholic compared to nonalcoholic cirrhotics (20.8 +/- 7.3 vs 16.4 +/- 6.3 mm Hg; P < 0.05), but this was not significant in multivariate analysis. The HVPG was comparable between hepatitis B- and hepatitis C virus-related cirrhotics (P = 0.8). Cirrhotics with ascites had a higher HVPG than those without ascites (18.5 +/- 5.6 vs 16.6 +/- 7.6 mm Hg; P = 0.02). In multivariate analysis, only Child's status, size of varices, and variceal bleed predicted higher HVPG. HVPG is higher in cirrhotics with large varices and a history of bleed. There is a good correlation between HVPG and large varices, bleeder status, and ascites. A higher HVPG reflects more severe liver disease. The etiology of liver disease did not influence the portal pressure. PMID- 17080246 TI - Studying the overlap between IBS and GERD: a systematic review of the literature. AB - Evidence points to a significant overlap between irritable bowel syndrome (IBS) and gastroesophageal reflux disease (GERD). In this study, we evaluate this overlap by conducting a systematic review of the literature. Six electronic databases from 1966 through January 2005 were screened by multiple search terms to identify all epidmiological evidence linking IBS and GERD. In addition, AGA meeting abstracts for 2003 and 2004 were also screened. All studies were validated by the authors and data extracted according to predefined criteria. As a separate search strategy, studies evaluating the prevalence of IBS and GERD in the general population were sought. These articles were obtained to compare the prevalence of IBS and GERD in the community to the degree of overlap. The search identified 997 original titles with 15 publications that fulfilled our eligibility criteria. Among the 15 studies, 7 determined the GERD maximum mean prevalence in patients already diagnosed with IBS to be 39.3% and the weighted mean 30.3%. The other 7 studies examined the prevalence of IBS in patients already diagnosed with GERD. The maximum mean prevalence of IBS in subjects with known GERD was 48.8% and the weighted mean 60.5%. Based on the prevalence of IBS (12.1%) and GERD (19.4%) in the community, the rate of IBS in the non-GERD community was calculated to be only 5.1%. There is a strong overlap between GERD and IBS that exceeds the individual presence of each condition. In the absence of GERD, IBS is relatively uncommon. PMID- 17080247 TI - Sleep-related autonomic disturbances in symptom subgroups of women with irritable bowel syndrome. AB - The objective was to investigate whether predominant symptom patterns in women with irritable bowel syndrome (IBS) affect autonomic activity during sleep. Seventy-five women with IBS underwent a polysomnographic sleep study. Twenty-two of the IBS patients were diarrhea-predominant (IBS-D), 33 were constipation predominant (IBS-C), and 20 patients had alternating symptoms (IBS-A). Autonomic activity was measured by heart rate variability. The IBS-D group had significant vagal withdrawal compared to the IBS-A group during REM and non-REM sleep (P < 0.05). The IBS-D symptom subgroup had significantly (P < 0.05) greater sympathetic dominance during non-REM than IBS-A patients. Lower abdominal pain correlated with sympathetic dominance during sleep in the IBS-D group (r=0.54, P < 0.01). The IBS-D patients were physiologically distinct with regard to autonomic functioning during sleep compared to the alternating patients, but not the constipated patients. Sleep appears to unmask differences in autonomic activity that may distinguish IBS patients. PMID- 17080248 TI - Low-dose naltreoxone for the treatment of irritable bowel syndrome: a pilot study. AB - Preclinical studies have shown that a very low dose of naltreoxone hydrochloride (NTX), an opiate antagonist, can block excitatory opioid receptors without affecting inhibitory opioid receptors, resulting in analgesic potency without side effects. The present study assessed the efficacy and safety of PTI-901 (low dose NTX) treatment in Irritable bowel syndrome (IBS) patients. Forty-two IBS patients participated in an open-label study. Participants received 0.5 mg PTI 901/day for 4 weeks and were evaluated during baseline, during treatment, and at 4-week follow-up. Patients recorded degree of abdominal pain, stool urgency, consistency, and frequency. Primary outcomes were number of pain-free days and overall symptom relief, evaluated by a global assessment score. Data were analyzed per protocol. Global assessment improved in 76% of 42 patients. During treatment, the mean weekly number of pain-free days increased from 0.5+/-1 to 1.25+/-2.14 (P=0.011). There were no significant adverse reactions. PTI-901 improves pain and overall feeling, and is well tolerated by IBS patients. A large, randomized, double-blind, placebo-controlled study is justified. PMID- 17080249 TI - Progression of atrophic gastritis and intestinal metaplasia drives Helicobacter pylori out of the gastric mucosa. AB - This study was performed to evaluate the implication of anti-H. pylori IgG positivity when CLOtest, histological test, and culture in the antrum and body are all negative, and to find out the specific disease category that is more affected by the hostile relationship of atrophic gastritis and intestinal metaplasia (IM) with H. pylori. Four hundred thirty-six patients (84 controls, 69 with duodenal ulcer, 96 with benign gastric ulcer, 43 with dysplasia, 144 with gastric cancer), who had not received any eradication therapy, were divided into three groups according to H. pylori test: CLOtest or histological H. pylori positive group (group A; 294 cases), only anti-H. pylori IgG-positive group (group B; 62 cases), and anti-H. pylori IgG-negative group (group C; 80 cases). The grade of neutrophil and monocyte infiltration, atrophic gastritis, and IM was compared according to the updated Sydney system classification. Neutrophil and monocyte infiltrations were significantly severe in the group A. In contrast, the grade of atrophic gastritis and IM in the antrum was significantly higher in group B than the other two groups, A or C. When patients were divided according to the disease outcome in each group, the grade of IM in the body was statistically higher only in the patients with cancer or dysplasia in group B. These results suggest that anti-H. pylori IgG positivity with all negative invasive H. pylori tests represents past infection with H. pylori rather than a false negative, especially in the case of dysplasia and gastric cancer. PMID- 17080250 TI - During thioacetamide-induced acute liver failure, the proliferative response of hepatocytes to thyroid hormone is maintained, indicating a potential therapeutic approach to toxin-induced liver disease. AB - In toxic liver injury, proliferation of preexisting hepatocytes helps restore liver mass and function. While loss of liver mass per se stimulates hepatocyte proliferation, exogenous mitogens have a potential role in enhancing liver regeneration. The aim of this study was to characterize the effects of the mitogen, tri-iodothyonine, on the regenerative capacity of hepatocytes during thioacetamide-induced liver failure. Rats received (two) thioacetamide injections and, 12 hr later, either tri-iodothyonine or vehicle-only control. Liver cell proliferation was assessed and comparison made with other control groups receiving tri-iodothyonine or vehicle only. In rats with thioacetamide-induced hepatitis the proportion of hepatocytes in S-phase was greater in the tri iodothyonine group (27+/-3.5%) compared to the vehicle-only group (20+/-2.5%; P < 0.05), with, notably, a greater number of midzonal (BrdU) positive hepatocytes in the tri-iodothyonine group. We conclude that the ability of hepatocytes in the midzonal areas of rat liver to proliferate in response to tri-iodothyonine is maintained during severe acute toxic injury. PMID- 17080251 TI - The use of early and midpoint adenoma-carcinoma sequence biomarkers in prediction of neoplastic progression in patients with a history of colorectal neoplasia. AB - Since significant neoplasia after initial colonoscopy is low, we conducted this pilot study to compare the predictive role for colorectal neoplasia recurrence of anti-DCC with that of Adnab-9 binding to colonic effluent of high-risk patients. DCC and Adnab-9 effluent ELISA were performed at baseline colonoscopies. The results of follow-up colonoscopies were reviewed. To ensure specificity, immunohistochemistry and Western blot was performed with anti-DCC and for Adnab-9 where optimal fixation times were also evaluated. Mean follow-up was 2.6 years. Of 21 patients, 6 of 10 who progressed to CRN and 2 of 11 who did not had a positive Adnab-9 ELISA result (P=0.08). Despite an initial good correlation with Adnab-9 ELISA results in a smaller dataset, we were unable to obtain consistent subsequent DCC immunohistochemistry or Western blot data using antibody from two different sources. However, the original dataset of Adnab-9 results was reproducible on repetition of the ELISA with a larger set of samples that included this initial dataset and optimal fixation time was 20 min. We conclude that Adnab-9 appears to be a promising prognostic marker for neoplasia in the high-risk population. Industry standards need to be developed for DCC monoclonal antibodies that may have similar utility. PMID- 17080252 TI - One-stage laparoscopic colorectal resection after placement of self-expanding metallic stents for colorectal obstruction: a prospective study. AB - The aim of this study was to assess the clinical outcomes of self-expandable metallic stents placing followed by laparoscopic resection and primary anastomosis for the treatment of acute colonic obstruction. From January 2003 to December 2004, 14 patients diagnosed with acute and complete colonic obstruction were treated with endoscopic colonic stenting as a bridge to an elective 1-stage laparoscopic resection. Three patients who underwent a successful stent insertion but had an inoperable tumor were excluded from the analyzed data. Ninety-three percent technical and clinical success was achieved. The stent insertion related perforation rate was 7% (1/14). The mean duration of stent insertion was approximately 1 hour and the mean time between the stent insertion and surgery was 6.2 days. Mean operating time was 132 +/- 38 minutes. No cases required conversion to laparotomy and there were no intraoperative complications. One case of anastomotic leakage was observed and treated by laparoscopic drainage and protective ileostomy. Ambulation time after operation was 1.8 +/- 0.6 days and total hospital stay length was 16.4 +/- 5.0 days. During a period of 11 +/- 7 months of follow-up, neither recurrences nor port-site metastases were observed. The management of acute colonic obstruction using endoscopic stent decompression, followed by laparoscopic resection, had good results and can be considered feasible and safe. Larger comparative studies may help to establish this approach. PMID- 17080253 TI - Endoscopic ultrasonography versus other diagnostic modalities in the diagnosis of choledocholithiasis. AB - Until recently, endoscopic retrograde cholangiopancreatography (ERCP) has been considered the gold standard for the diagnosis of and therapy in patients with suspected choledocholithiasis. However, the non-negligible complication rate of diagnostic and therapeutic ERCP has led investigators to identify different noninvasive diagnostic modalities. Endoscopic ultrasonography has been proved to be of great sensitivity (up to 97%) in the diagnosis of even tiny stones that can be easily masked by contrast medium during ERCP, without any procedure-related complications and with a negative predictive value reaching 100%, meaning that it can accurately and safely identify patients with choledocholithiasis, thereby avoiding inappropriate instrumental exploration of the common bile duct. PMID- 17080254 TI - Effects of L-acetylcarnitine on cirrhotic patients with hepatic coma: randomized double-blind, placebo-controlled trial. AB - Multiple therapeutic modalities have been used to treat hepatic encephalopathy. L: -Acetylcarnitine (LAC) is a physiologically active substance that improves both the energetic and the neurotransmission profiles. LAC is able to cross the hematoencephalic barrier and reach the cerebral regions, where the acetylic group may be utilized. The aim of this work was to evaluate the efficacy of LAC in the treatment of hepatic coma in cirrhotic patients. Twenty-four suitably selected patients were enrolled in the study and, following randomization, received either LAC (n=13) or placebo (n=11). Statistically significant differences in neurological findings, as evaluated by the Glasgow Scale, as well as in ammonia serum levels and BUN were found following LAC treatment. In the placebo group we observed two cases of improved neurological findings as well as one case of improved EEG grading. In the other group we observed an improvement of neurological findings and of EEG grade in 10 and 8 subjects, respectively. Noteworthily, seven (54%) patients went from grade 4 down to grade 3, and one from grade 4 down to grade 1. The improvement in the neurological picture was evident at between 1 and 4 hr after the end of treatment, remaining until 24 hr after. No side effects were observed in our study series. Our study demonstrates that LAC administration improved neurological and biohumoral symptoms in selective cirrhotic patients with hepatic coma. PMID- 17080255 TI - Anti-inflammatory effect of bioflora probiotic administered orally or subcutaneously with live or dead bacteria. AB - The anti-inflammatory effect of Bioflora probiotic administered orally or subcutaneously with viable or nonviable bacteria was analyzed in two experimental models of randomly selected female Wistar rats. The use of indomethacin at a dose of 50 mg/kg was associated with gastric necrotic lesions and multiple erosions of the small intestine, with marked mucosal neutrophil infiltrate measured by myeloperoxidase (MPO). Probiotics prevented both gastrointestinal lesions and neutrophil infiltrate (p < 0.001). In the model of carrageenin-induced plantar edema in the rat, the oral or subcutaneous administration of Bioflora with live or dead bacteria proved to prevent plantar edema (p < 0.001). We concluded that Bioflora probiotic given orally or subcutaneously with live or dead bacteria has an anti-inflammatory effect. PMID- 17080256 TI - Phase II clinical and pharmacologic study of radiation therapy and carboxyamido triazole (CAI) in adults with newly diagnosed glioblastoma multiforme. AB - INTRODUCTION: Carboxyamido-triazole (CAI) is a synthetic inhibitor of non-voltage gated calcium channels that reversibly inhibits angiogenesis, tumor cell proliferation, and metastatic potential. This study examined the efficacy, safety and pharmacokinetics of oral CAI in the treatment of patients with newly diagnosed glioblastoma multiforme (GBM) in an open-label, single arm non randomized phase 2 trial. METHODS: Eligible patients with histologically confirmed GBM started CAI therapy (250 mg daily) on the first day of radiation (6000 cGy in 30 fractions) and continued until progression, unless side effects became intolerable. The primary outcome was survival compared to historical controls within the NABTT CNS Consortium database. Secondary outcomes included toxicity and pharmacokinetic parameters. RESULTS: Fifty-five patients were enrolled with a median Karnofsky performance status of 90 and age of 56 years. Forty-six (84%) of these patients had debulking surgeries and 52 have died. The median survival was 10.3 months (95% confidence interval (CI), 8.5-12.8) compared to 12.1 months (95% CI, 10.3-13.3) in the NABTT reference group (p = 0.97). Significant toxicities included 2 incidents of reversible vision loss. The mean CAI plasma concentration for patients taking enzyme inducing antiepileptic drugs (EIAED) was 1.35 +/-1.22 compared to 4.06 +/- 1.50 (p < 0.001) for subjects not taking these agents. Overall survival and grade > or = 3 toxicities were comparable by EIAED status. CONCLUSIONS: This study demonstrated that (1) CAI can be administered safely with concomitant cranial irradiation, (2) the pharmacokinetics of CAI are significantly affected by co-administration of EIAED, and (3) the survival of patients with newly diagnosed GBM was not improved with this novel agent, despite achieving adequate drug levels. PMID- 17080257 TI - Asymmetric specialization and extinction risk in plant-flower visitor webs: a matter of morphology or abundance? AB - A recently discovered feature of plant-flower visitor webs is the asymmetric specialization of the interaction partners: specialized plants interact mainly with generalized flower visitors and specialized flower visitors mainly with generalized plants. Little is known about the factors leading to this asymmetry and their consequences for the extinction risk of species. Previous studies have proposed random interactions proportional to species abundance as an explanation. However, the simulation models used in these studies did not include potential biological constraints. In the present study, we tested the potential role of both morphological constraints and species abundance in promoting asymmetric specialization. We compared actual field data of a Mediterranean plant-flower visitor web with predictions of Monte Carlo simulations including different combinations of the potential factors structuring the web. Our simulations showed that both nectar-holder depth and abundance were able to produce asymmetry; but that the expected degree of asymmetry was stronger if based on both. Both factors can predict the number of interaction partners, but only nectar-holder depth was able to predict the degree of asymmetry of a certain species. What is more, without the size threshold the influence of abundance would disappear over time. Thus, asymmetric specialization seems to be the result of a size threshold and, only among the allowed interactions above this size threshold, a result of random interactions proportional to abundance. The simulations also showed that asymmetric specialization could not be the reason that the extinction risk of specialists and generalists is equalized, as suggested in the literature. In asymmetric webs specialists clearly had higher short-term extinction risks. In fact, primarily generalist visitors seem to profit from asymmetric specialization. In our web, specialists were less abundant than generalists. Therefore, including abundance in the simulation models increased the difference between specialists and generalists even more. PMID- 17080258 TI - Inheritance of organelle DNA markers in a pea cross associated with nuclear cytoplasmic incompatibility. AB - An unusual biparental mode of plastid inheritance was found in pea, in a cross associated with nuclear-cytoplasmic incompatibility manifested as deficiency of chlorophyll pigmentation. Plastid DNA marker trnK and mitochondrial DNA marker cox1 were analyzed in F1 progeny that received cytoplasm from an accession of a wild subspecies Pisum sativum ssp. elatius. Plants with sectors of green tissue on leaves and seed cotyledons with green patches on an otherwise chlorotic background were found to carry paternally inherited plastid DNA, suggesting that photosynthetic function was affected by nuclear-cytoplasmic conflict and required proliferation of paternally inherited plastids for normal performance. The paternally inherited plastid DNA marker was also observed in the roots. The presence of the paternal marker in cotyledons, roots and leaves was independent of each other. Inheritance of the mitochondrial DNA marker cox1 appeared to be of the maternal type. PMID- 17080259 TI - The effect of stimulation technique on sympathetic skin responses in healthy subjects. AB - The aim of this study was to collect normative data for sympathetic skin responses (SSR) elicited by electrical stimulus of the ipsilateral and contralateral peripheral nerves, and by magnetic stimulus of cervical cord. SSRs were measured at the mid-palm of both hands following electrical stimulation of the left median nerve at the wrist and magnetic stimulation at the neck in 40 healthy adult volunteers (mean age 52.2 +/- 12.2 years, 19 males). The onset latency, peak latency, amplitude and area were estimated in "P" type responses (i.e., waveforms with a larger positive, compared to negative, component). SSR onset and peak latency were prolonged when the electrical stimulus was applied at the contralateral side (i.e., the SSR recorded in the right palm P < 0.001). The onset latency was similar on both sides during cervical magnetic stimulation. However, peak latency was faster on the left side (P < 0.03). Comparison of electrical and magnetic stimulation revealed that both the onset and peak latency were shorter with magnetic stimulation (P < 0.001). The latency of a SSR varies depending on what type of stimulation is used and where the stimulus is applied. Electrically generated SSRs have a longer delay and the delay is prolonged at the contralateral side. These factors should be taken into account when interpreting SSR data. PMID- 17080260 TI - Selective bronchodilatory effect of Rooibos tea (Aspalathus linearis) and its flavonoid, chrysoeriol. AB - BACKGROUND: Rooibos tea (Aspalathus linearis) is commonly used for hyperactive gastrointestinal, respiratory and cardiovascular disorders. AIM OF STUDY: The aqueous extract of Rooibos tea (RT) was studied for the possible bronchodilator, antispasmodic and blood pressure lowering activities in an attempt to rationalize some of its medicinal uses. METHODS: Isolated tissue preparations, such as rabbit jejunum, aorta and guinea-pig trachea and atria were set up in appropriate physiological salt solutions and aerated with carbogen. For in vivo studies rats were anesthetized with pentothal sodium and blood pressure was measured through carotid artery cannulation. RESULTS: In jejunum, RT caused a concentration dependent relaxation of low K(+) (25 mM)-induced contractions, with mild effect on the contractions induced by high K(+) (80 mM). In presence of glibenclamide, the relaxation of low K(+)-induced contractions was prevented. Similarly, cromakalim caused glibenclamide-sensitive inhibition of low K(+), but not of high K(+), while verapamil did not differentiate in its inhibitory effect on contractions produced by the two concentrations of K(+). Like in jejunum, RT caused glibenclamide-sensitive relaxation of low K(+)-induced contractions in trachea and aorta, but with a 20 times higher potency in trachea. In atria, RT was least potent with weak inhibitory effect on atrial force and rate of contractions. RT caused a dose-dependent fall in arterial blood pressure in rats under anesthesia. Among the tested pure compounds of Rooibos, chrysoeriol showed selective bronchodilator effect. Chrysoeriol (luteolin 3'-methyl ether) is a bioactive flavonoid known for antioxidant, antiinflammatory, antitumor, antimicrobial, antiviral, and free radical scavenging activities. CONCLUSION: These results indicate that the bronchodilator, antispasmodic and blood pressure lowering effects of Rooibos tea are mediated predominantly through K(ATP) channel activation with the selective bronchodilatory effect. This study provides a sound mechanistic basis for the wide medicinal use of Rooibos tea, with the therapeutic potential to be developed for congestive respiratory ailments. PMID- 17080261 TI - Genotype-by-nutrient interactions assessed in European obese women. A case-only study. AB - BACKGROUND: The development of obesity is influenced by both genetic and environmental risk factors. Whereas changes in the environment appear to be responsible for the increasing prevalence of obesity, genetic factors interacting with environmental factors would contribute to explain obesity onset and severity. AIM: To explore epidemiologic genotype-by-nutrient interactions in obesity. METHODS: A total of 42 polymorphisms of 26 candidate genes for obesity were genotyped in 549 adult obese women recruited from eight European centres in a case-only study. The nutritional variables assessed in this study were the dietary fibre intake (grams per day), the ratio of dietary polyunsaturated fat to saturated fat (P:S ratio) and the percentage of energy derived from fat in the diet as calculated from a weighed three-day food record (%E). Under the assumption of genotype-nutrient independence in the population, the odds ratio calculated in a sample of obese women would indicate the existence of genotype-by nutrient interactions, measured as deviations from the multiplicative effects of the genetic and the nutrient factors separately. RESULTS: No new but confirmaty evidences for genotype-by-nutrient interactions in obesity were detected in this case-only study. The test of interaction between fibre intake and the -514 C > T polymorphism of the hepatic lipase gene (LIPC) yielded P-values of 0.01 across different statistical models. Likewise, the -11377G > C polymorphism of the adiponectin gene (ADIPOQ) and the -681 C > G polymorphism of the PPARG3 gene might interact with the percentage of energy derived from fat in the diet for the development of obesity (P-values in the range of 0.01-0.05 across different statistical models). The P-values were not adjusted for multiple testing, so these results should be considered with caution. CONCLUSIONS: Although the use of obese-only samples is theoretically a useful approach to detect interactions, few genotype-by-nutrient interactions have been suggested in obese European women after the analysis of candidate polymorphisms and the selected nutrient variables. The most remarkable multiplicative interaction found in this study refers to the combination of the hepatic lipase gene polymorphism -514 C > T and fibre intake. PMID- 17080262 TI - Collagen in colon disease. AB - The pathophysiology of wound healing in the bowel wall suggests that collagen and matrix metalloproteinases (MMPs) have an important role in the changes of the bowel wall seen in several colonic diseases. Several recent studies suggest that disturbances of the collagen texture and the extracellular matrix (ECM) metabolism are major factors leading to the onset of diverticular disease. Changes of the ECM also play a role in the development of inflammatory bowel diseases. Regarding the permanent remodeling of the bowel wall, any imbalance of the ECM could support the onset of chronic inflammation and the development of fistula formation, such as that seen in patients with Crohn's disease. Disturbances of the ECM play a role in the pathogenesis of anastomotic leakage after large bowel surgery and suggest the presence of a genetically defined risk population with disturbed wound healing mechanisms. This concept could explain the well known situation where an anastomotic breakdown is observed, despite the absence of other known risk factors and after a technically correct anastomosis. PMID- 17080263 TI - Hypophysitis. AB - Hypophysitis is an uncommon inflammatory condition that may affect the pituitary gland and stalk. Patients often present with varying degrees of hypopituitarism. The diagnosis is often made presumptively based on clinical history and biochemical data but may also be supported by magnetic resonance imaging. Therapy is generally supportive in nature but may require surgery for pathological diagnosis and treatment of mass effect. PMID- 17080264 TI - Neurogenic diabetes insipidus. PMID- 17080265 TI - Craniopharyngioma. AB - Craniopharyngiomas are benign tumors of the parasellar region. These tumors may be cystic, solid, or combinations of the two. They have a bimodal age distribution and no apparent gender predilection. Patients may present with endocrinopathy or symptoms related to mass effect from the growing tumor. Patients require a multidisciplinary approach during their diagnostic evaluation and subsequent to initiating therapy. Endocrinopathy should be recognized and treated with appropriate hormonal replacement. Surgery is the first-line therapy for most patients. The specific surgical approach must be tailored to the specific clinical situation and depends on the patient age, endocrine status, and the geometry and consistency of the tumor. Whereas most solid tumors will require craniotomy or transsphenoidal surgery, some cystic tumors may be adequately managed with intracavitary therapies. Subtotally resected or residual tumors often require adjuvant radiation therapy or radiosurgery. Long-term multidisciplinary follow-up is necessary for all patients. PMID- 17080266 TI - TSH adenomas. AB - Thyrotropin (TSH) adenomas are uncommon tumors of the anterior pituitary. Clinically, they may present with hyperthyroidism, hypothyroidism, hypopituitarism, or visual loss. Proper diagnosis requires appropriate neuroendocrine evaluation. Although transsphenoidal surgery remains the first line treatment, adjuvant medical therapy is frequently required for these often invasive tumors. Radiotherapy or radiosurgery should be considered for those patients with persistent TSH hypersecretion following surgery and medical therapy. The clinical evaluation and treatment algorithm for patients with TSH adenomas is discussed. PMID- 17080267 TI - Analysis of respiratory complications after minimally invasive esophagectomy: preliminary observation of persistent aspiration risk. AB - Minimally invasive (MI) esophageal resection (ER) has the theoretical advantage of reduced postoperative complications compared with standard ER. However, the impact of MIER on rates and severity of pulmonary complications is unclear. Four patients underwent laparoscopic gastroesophageal mobilization and resection followed by gastric pull-up and cervical esophageal anastomosis (MIER). Videofluoroscopic swallowing studies (VFSS) assessed pharyngolaryngeal function postoperatively. All postoperative complications were documented. Each MIER was completed successfully without intraoperative complications. Mean operative time was 4.3 +/- 2 h. Postoperatively, VFSS detected laryngeal penetration, vocal cord paralysis, and/or aspiration in three patients, two of whom experienced severe respiratory complications. MIER patients are susceptible to aspiration, likely due to transient denervation of the pharynx and laryngeal structures. Following MIER, aggressive pulmonary toilet and aspiration precautions are emphasized to reduce pulmonary complications. Furthermore, serial evaluation of deglutition is encouraged to guide the safe and appropriate resumption of oral feeding. PMID- 17080268 TI - Letter to the Editor: Comment on "The Adult Asperger Assessment (AAA): a diagnostic method", Simon Baron-Cohen, Sally Wheelwright, Janine Robinson and Marc Woodbury-Smith, Journal of Autism and Developmental Disorders Vol 35(6). PMID- 17080269 TI - Response to "Letter to the Editor: The overlap between alexithymia and Asperger's syndrome", Fitzgerald and Bellgrove, Journal of Autism and Developmental Disorders, 36(4). PMID- 17080270 TI - Validity of the social communication questionnaire in assessing risk of autism in preschool children with developmental problems. AB - This study estimates the sensitivity and specificity of the social communication questionnaire (SCQ) for autistic spectrum disorders in preschool children at high risk for developmental problems referred to a tertiary centre and compares the predictions of the SCQ and the referrer. The SCQ was completed by 81 parents prior to multidisciplinary assessment and compared with the final diagnosis. The sensitivity and specificity were 93% and 58% for children aged 2-6, and 100 and 62% for children aged 3-5 years, with a cut-off score of 11. The SCQ performed better than referrers. Low-specificity means the SCQ is not suitable as a diagnostic tool but will assist clinicians and tertiary services in selecting children with developmental problems who require autism-specific assessment. PMID- 17080271 TI - Adults with autism show increased sensitivity to outcomes at low error rates during decision-making. AB - Decision-making is an important function that can be quantified using a two choice prediction task. Individuals with Autistic Disorder (AD) often show highly restricted and repetitive behavior that may interfere with adaptive decision making. We assessed whether AD adults showed repetitive behavior on the choice task that was unaffected by changing task demands, by examining the influence of experimenter-determined error rates on decision-making. Sixteen AD adults and 14 typically developed subjects were administered a two-choice task using three error rate conditions. Although AD subjects showed occurrences of stereotyped responding, their decision-making behavior was strongly affected by changes in task demands, especially when they experienced frequent success. Thus, behavioral paradigms that provide frequent reinforcement may be helpful in modifying decision-making abilities in AD. PMID- 17080272 TI - Adults' versus children's perceptions of a child with autism or attention deficit hyperactivity disorder. AB - The present study examined public perceptions toward children with autism or with attention deficit hyperactivity disorder (ADHD). A convenience sample was used consisting of 30 children (7-12-year-olds) and 30 adults. Participants read a stereotyped scenario featuring either a child with autism, a child with ADHD, or a normal child. Child participants were significantly more likely than adults to (a) express dislike/avoidance toward a child described with either stereotypic autistic or ADHD behaviors, and (b) perceive the child with ADHD as unlike themselves. However, child participants and adults were equally likely to see the autistic child as unlike themselves. Reasons for the different perceptions of children and adults may include differences in perceived threat and in categorization. PMID- 17080273 TI - Clarifying the associations between language and social development in autism: a study of non-native phoneme recognition. AB - Autism spectrum disorders (ASDs) are characterized by correlated deficiencies in social and language development. This study explored a fundamental aspect of auditory information processing (AIP) that is dependent on social experience and critical to early language development: the ability to compartmentalize close sounding speech sounds into singular phonemes. We examined this ability by assessing whether close-sounding non-native language phonemes were more likely to be perceived as disparate sounds by school-aged children with high-functioning ASD (n = 27), than by unaffected control subjects (n = 35). No significant group differences were observed. Although earlier in autistic development there may exist qualitative deficits in this specific aspect of AIP, they are not an enduring characteristic of verbal school-aged children with ASD. PMID- 17080274 TI - Individual and marital adaptation in men with autism spectrum disorder and their spouses: the role of social support and coping strategies. AB - The aim of the present study was to examine the predictive value of social support and coping for individual and marital adaptation in adult men with autism spectrum disorder (ASD) and their spouses, based on the double ABCX model of adaptation. Twenty-one couples participated in the study and completed measures of stressor severity, social support, coping, individual and marital adaptation. Bivariate analyses showed that each of the model components was related to adaptation in men and women. Hierarchical regression analyses revealed that, after controlling for relevant demographics and stressor severity, informal support was a strong, and unique predictor of adaptation in both spouses (explained variance: 27-89%). Coping did not add to the prediction of adaptation. Clinical implications and limitations are discussed. PMID- 17080275 TI - Atypical lexical/semantic processing in high-functioning autism spectrum disorders without early language delay. AB - Although autism is associated with impaired language functions, the nature of semantic processing in high-functioning pervasive developmental disorders (HFPDD) without a history of early language delay has been debated. In this study, we aimed to examine whether the automatic lexical/semantic aspect of language is impaired or intact in these population. Eleven individuals with Asperger's Disorder (AS) or HFPDD-Not Otherwise Specified (NOS) and age-, IQ-, and gender matched typically developing individuals performed a semantic decision task in four conditions using an indirect priming paradigm. Semantic priming effects were found for near-semantically related word pairs in the controls, whereas this was not the case in the AS or HFPDDNOS participants. This finding suggests similarities in the underlying semantic processing of language across PDD subtypes. PMID- 17080276 TI - Bacterial endocarditis following rubber band ligation in a patient with a ventricular septal defect: report of a case and guideline analysis. AB - Rubber band ligation is a common option used to treat symptomatic internal hemorrhoids. Severe complications such as pelvic sepsis are a rare occurrence. We report a case of endocarditis leading to septic pulmonary and renal emboli following single-quadrant rubber band ligation. The patient had a known ventricular septal defect and developed low back pain and fever after ligation of a right anterior internal hemorrhoid. He was found to have septic pulmonary emboli, a renal wedge septic infarct, and a large vegetation on his membranous ventricular septal defect requiring operative intervention. Before this report, rubber band ligation has not been associated with endocarditis. According to several guidelines, this patient did not require antibiotic prophylaxis. It is unclear whether prophylaxis could have prevented this complication. Surgeons utilizing rubber band ligation need to be familiar with all potential complications. PMID- 17080277 TI - Perianal local block for stapled anopexy. AB - PURPOSE: This study was designed to demonstrate the usefulness of a method of regional anesthesia for circular stapler anopexy for prolapsing hemorrhoids. METHODS: Thirty-three patients consented to stapled anopexy under perianal local anesthesia. Eighteen patients with stapled anopexy under general anesthesia were controls. The perianal block was applied with 40 ml of ropivacaine, 4.75 mg/ml, injected immediately peripheral to the external sphincter. A submucosal block with 15 ml of ropivacaine, 2 mg/ml, was added after applying the pursestring suture. Postoperative pain was rated by the patient for 14 days by using a ten point visual analogue scale. Patients also submitted a preoperative and postoperative (3-6 months) symptom questionnaire to rate anal symptoms. RESULTS: No operation was converted to general anesthesia. Operation time was similar in both groups. All patients in the local anesthesia group were pain free at discharge. The sums of pain scores during 14 days for daily average pain and peak pain were similar in both groups (average pain 23 (local anesthesia) vs. 35 (general anesthesia); peak pain 39 (local anesthesia) vs. 50 (general anesthesia); P>0.05). The preoperative symptom scores were 7.8 (local anesthesia) vs. 8.9 (general anesthesia) points, and the follow-up scores were 2.2 (local anesthesia) and 2.7 (general anesthesia), a significant improvement (P=0.001) in both groups but not different between groups. CONCLUSIONS: A perianal local block is easy to apply and has a high degree of acceptance among patients. The operation time, postoperative pain, and success rates of the operation equaled those of stapled anopexy performed under general anesthesia. The advantages are quicker turnover between cases and simpler management of pain-free postoperative patients in day surgery. PMID- 17080278 TI - Sacral nerve stimulation in fecal incontinence: are there factors associated with success? AB - PURPOSE: Sacral nerve stimulation has been used successfully in treating fecal incontinence. This study was designed to evaluate the proportion of patients with unsuccessful implantation despite positive test stimulation and to examine and compare factors associated with the success of the transitory and permanent sacral nerve stimulation. METHODS: A total of 61 patients (55 females; median age, 56 (range, 33-77) years) with refractory fecal incontinence underwent temporary stimulation. A 50 percent or greater improvement in the number of episodes of fecal incontinence or urgency was required to proceed to permanent implantation and was the criteria of success of permanent sacral nerve stimulation at the last follow-up visit in implanted patients. The factors compared between the success and the failure groups during temporary and permanent stimulation were patients' age and gender, diagnosis and characteristics of fecal incontinence, previous surgery, quality of life scores, anorectal manometry, endoanal ultrasound, and electrophysiologic tests performed before stimulation. RESULTS: Temporary stimulation was successful in 35 patients (57.4 percent). A permanent neurostimulation device was implanted in 33 patients. Age was the only factor related to success of the temporary stimulation (P=0.03). After permanent implantation, 31 percent of patients did not attain screening phase results for the number of episodes of fecal incontinence or urgency. A neurologic disorder was more frequently the origin of fecal incontinence in the success group compared with others (P=0.03). The left bulbocavernosus reflex was more frequently delayed in the success group than in the others (P=0.03), and a prolonged or absent bulbocavernosus reflex was more frequent in the success group than in the failure group (P=0.03). CONCLUSIONS: Patients with fecal incontinence from neurologic origins could be good candidates for sacral nerve stimulation. PMID- 17080279 TI - Tumor invasion of lymph node capsules in patients with Dukes C colorectal adenocarcinoma. AB - PURPOSE: The objective of this study was to investigate the correlation between the microscopic findings of positive lymph nodes, especially focusing on capsular invasion, and the outcome after curative surgical resection of colorectal cancer. METHODS: We analyzed 480 positive lymph nodes from 155 consecutive patients with Stage III colorectal cancer to determine the frequency and significance of lymph node capsular invasion. Recurrence-free and cancer-specific survival rates were assessed in the patients with and without lymph node capsular invasion. RESULTS: Between April 1995 and December 2000, 406 consecutive patients with primary colorectal cancer underwent curative resection. Regional lymph node metastases were present in 155 cases (38.2 percent). During the median follow-up period of 4.8 years, 41 patients (26.5 percent) developed recurrent disease and 28 patients died of cancer. Lymph node capsular invasion was detected in one or more lymph nodes from 75 cases (48.3 percent). The five-year recurrence-free rate was 56.1 percent in this group, whereas in the 80 patients without lymph node capsular invasion the rate was 88 percent (P<0.01). Features that were associated with recurrent disease were greater number of positive lymph nodes, venous invasion in primary tumor, infiltrative growth pattern of intranodal tumor, and presence of lymph node capsular invasion. Multivariate analysis identified lymph node capsular invasion as the only significant prognostic factor for recurrence. In multivariate analysis with regard to survival, lymph node capsular invasion, venous invasion, and number of positive nodes remained as significant prognostic factors. CONCLUSIONS: Lymph node capsular invasion, determined by routine hematoxylin-eosin staining, is a potent prognostic factor in Stage III colorectal cancer. PMID- 17080280 TI - Sonographic characteristics of rectal sensations in healthy females. AB - PURPOSE: This study was designed to characterize rectal sensations by visualizing the internal and external anal sphincter and intra-anal transport of bolus during elicited rectal sensations. METHODS: The anal canal was visualized with real-time transperineal ultrasonography in 13 healthy female volunteers. Rectal sensations were elicited by injecting water into the rectum. The ultrasound images were recorded on a videotape and analyzed offline. RESULTS: The median time between an injection of water and the events studied was calculated in 105 rectal sensations. A relaxation in the internal anal sphincter (4 seconds after the injection of water), an antegrade transport of bolus (4 seconds) into the anal canal, and a contraction in the external anal sphincter (5 seconds) were observed before a sensation (6 seconds) was reported. The antegrade flow continued until the distal internal anal sphincter contracted (18 seconds) and the bolus moved in a retrograde transport direction (17 seconds) thereafter the sensation disappeared (18 seconds) and the external anal sphincter relaxed (22 seconds). A significant correlation in time between the end of the sensation, contraction in the internal anal sphincter, reversed flow of anal contents, and relaxation of the external anal sphincter was found (Pearson, P<0.01). CONCLUSIONS: The results verified that the internal anal sphincter contributes to the perception of rectal sensations by a relaxation allowing intra-anal bolus to increase the pressure on the anoderm during rectal contraction. A new observation is presented on the time relation between contraction in the distal internal anal sphincter, reversed flow in the anal canal, and the end of rectal sensations. PMID- 17080281 TI - Comparison of outcomes in Z-plasty and delayed healing by secondary intention of the wound after excision of the sacral pilonidal sinus: results of a randomized, clinical trial. AB - PURPOSE: Pilonidal sinus is a common disease, mostly among young males. Although the conventional surgery, secondary repair of the wound after wide excision with or without curettage, is not technically difficult, it takes a long time, weeks or months, for a wound to heal. In this study, we compared outcomes and complications of Z-plasty repair with outcomes and complications of delayed healing by secondary intention (conventional surgery). METHODS: In a randomized, clinical trial, we recruited 72 patients in each arm. All of the patients were operated on by a general surgeon and were followed up for variable periods. Patients in the Z-plasty arm were followed for a mean period of 22.01 months. Patients conventional surgery arm were followed for a mean period of 22.23 months. The main studied outcomes were healing of the wound and disease recurrence during the follow-up period. The main complications that were taken into account were bleeding, hematoma, infection, and recurrence. RESULTS: The two arms were comparable in the age and gender distributions, duration of disease, and history of any types of operations. The duration of hospitalization after the operation was shorter in the conventional surgery group (1.76+/-0.75 days vs. 2.86+/-0.73 days, P<0.001). However, wounds healed much faster in the Z-plasty group (15.4 vs. 41 days, P<0.001). Moreover, it took longer for patients in the conventional surgery group to return to normal activity (17.5 vs. 11.9 days, P<0.001). There was no difference in postoperative complications regarding bleeding, hematoma, infection, or recurrence during the follow-up period in two arms. CONCLUSIONS: Healing was considerably faster in patients who were treated with Z-plasty technique of wound closure. This procedure can be used as a desirable treatment for pilonidal sinus, which is a noncomplex method without increased rate of complications. Nonetheless, it might slightly increase the duration of hospitalization. PMID- 17080282 TI - Solitary rectal ulcer syndrome in childhood: a rare, benign, and probably misdiagnosed cause of rectal bleeding. Report of three cases. AB - Solitary rectal ulcer syndrome is an uncommon and often underdiagnosed condition that usually presents with hematochezia, mucous discharge, and tenesmus. Its etiology is unknown but it seems related to excessive straining with defecation. Prolonged efforts force the anterior rectal mucosa into the anal canal with strangulation and appearance of congestion, edema, and ulceration. Histologic findings (fibromuscular obliteration of lamina propria and disorientation of muscle fibers) are characteristic, which helps to differentiate these lesions from other rectal entities. Although solitary rectal ulcer syndrome is rarely reported in children, it must be suspected in patients with rectal discharge of blood and mucus and previous disorders of evacuation. We present three children (aged 9, 10, and 14 years) with solitary rectal ulcer syndrome that had presented with rectal bleeding. A careful inquiry about evacuation habits and a high index of suspicion in children presenting with hematochezia helps to diagnose this possibly unrecognized or misdiagnosed entity in children. Endoscopy and histologic examination confirms this condition. PMID- 17080283 TI - Application of 10 percent formalin for the treatment of radiation-induced hemorrhagic proctitis. AB - PURPOSE: Rectal instillation of 4 percent formalin solution has been described as a successful treatment for hemorrhagic radiation proctitis recalcitrant to medical treatment. We present our experience with a new method of treatment involving the topical application of 10 percent buffered formalin, which is well tolerated and suitable for office use. METHODS: Patients with marked or refractory rectal bleeding and clinical features consistent with radiation proctitis were reviewed. Treatment involved direct application of a 10 percent buffered formalin solution to the affected mucosa using a 16-inch cotton tip applicator applied through a proctoscope in the office setting. RESULTS: A total of 100 patients with a mean age of 75 (range, 49-91) years were followed for 18 (range, 1-79) months. The interval from radiation exposure to formalin treatment was 21 months. Overall, 93 percent of patients had cessation of bleeding after an average of 3.5 formalin applications at two-week to four-week intervals. Patients with severe (Grade 3) proctitis and those taking aspirin required an average of 1.5 additional treatments. A total of eight patients rebled at a mean of 24 months from treatment; however, all responded to further applications of formalin. Three patients complained of anal pain and one experienced dizziness postprocedure for a complication rate of 1.1 percent. CONCLUSIONS: We present a simple, cost-effective, and well-tolerated method of controlling hemorrhagic radiation proctitis. It is performed by using materials readily available in the office of a colon and rectal surgeon, eliminating the need for bowel preparation, anesthesia, or a surgical suite. PMID- 17080284 TI - Anti-Saccharomyces cerevisiae antibody in intestinal Behcet's disease patients: relation to clinical course. AB - PURPOSE: This study was designed to assess anti-Saccharomyces cerevisiae antibody positive rate in Behcet's disease and intestinal Behcet's disease and to evaluate whether anti-Saccharomyces cerevisiae antibody expression is associated with clinical findings at diagnosis and clinical course of intestinal Behcet's disease. METHODS: One hundred six patients with intestinal Behcet's disease, 30 patients with Behcet's disease, and 45 healthy control subjects were included. Anti-Saccharomyces cerevisiae antibody was detected by indirect immunofluorescence assay. According to anti-Saccharomyces cerevisiae antibody expression, the various parameters at diagnosis, cumulative relapse rates, and cumulative probabilities of operation were analyzed. RESULTS: Anti-Saccharomyces cerevisiae antibody positive rate was 44.3 percent in intestinal Behcet's disease, 3.3 percent in Behcet's disease, and 8.8 percent in healthy control subjects. In patients with intestinal Behcet's disease, age, gender, distribution of Behcet's disease subtype, symptoms, laboratory tests, and colonoscopic findings at diagnosis were not different according to anti-Saccharomyces cerevisiae antibody expression. Cumulative probability of a first operation was significantly higher in anti-Saccharomyces cerevisiae antibody (+) intestinal Behcet's disease than in anti-Saccharomyces cerevisiae antibody (-) intestinal Behcet's disease: 44.8 and 17.2 percent at one year, and 53 and 24.3 percent at two years after diagnosis, respectively (P=0.006). The number of patients who underwent two or more operations was higher in anti-Saccharomyces cerevisiae antibody (+) intestinal Behcet's disease than in anti-Saccharomyces cerevisiae antibody (-) intestinal Behcet's disease (21.3 vs. 8.5 percent). The cumulative relapse rates were not different between the two groups. CONCLUSIONS: Anti Saccharomyces cerevisiae antibody positive rate was 44.3 percent in intestinal Behcet's disease. Clinical findings at diagnosis and cumulative relapse rates of intestinal Behcet's disease were not found to be associated with anti Saccharomyces cerevisiae antibody expression. However, patients with anti Saccharomyces cerevisiae antibody (+) intestinal Behcet's disease were more likely to receive surgical treatment. PMID- 17080285 TI - An extended RNA code and its relationship to the standard genetic code: an algebraic and geometrical approach. AB - An algebraic and geometrical approach is used to describe the primaeval RNA code and a proposed Extended RNA code. The former consists of all codons of the type RNY, where R means purines, Y pyrimidines, and N any of them. The latter comprises the 16 codons of the type RNY plus codons obtained by considering the RNA code but in the second (NYR type), and the third, (YRN type) reading frames. In each of these reading frames, there are 16 triplets that altogether complete a set of 48 triplets, which specify 17 out of the 20 amino acids, including AUG, the start codon, and the three known stop codons. The other 16 codons, do not pertain to the Extended RNA code and, constitute the union of the triplets YYY and RRR that we define as the RNA-less code. The codons in each of the three subsets of the Extended RNA code are represented by a four-dimensional hypercube and the set of codons of the RNA-less code is portrayed as a four-dimensional hyperprism. Remarkably, the union of these four symmetrical pairwise disjoint sets comprises precisely the already known six-dimensional hypercube of the Standard Genetic Code (SGC) of 64 triplets. These results suggest a plausible evolutionary path from which the primaeval RNA code could have originated the SGC, via the Extended RNA code plus the RNA-less code. We argue that the life forms that probably obeyed the Extended RNA code were intermediate between the ribo-organisms of the RNA World and the last common ancestor (LCA) of the Prokaryotes, Archaea, and Eucarya, that is, the cenancestor. A general encoding function, E, which maps each codon to its corresponding amino acid or the stop signal is also derived. In 45 out of the 64 cases, this function takes the form of a linear transformation F, which projects the whole six-dimensional hypercube onto a four-dimensional hyperface conformed by all triplets that end in cytosine. In the remaining 19 cases the function E adopts the form of an affine transformation, i.e., the composition of F with a particular translation. Graphical representations of the four local encoding functions and E, are illustrated and discussed. For every amino acid and for the stop signal, a single triplet, among those that specify it, is selected as a canonical representative. From this mapping a graphical representation of the 20 amino acids and the stop signal is also derived. We conclude that the general encoding function E represents the SGC itself. PMID- 17080287 TI - Hypothetical approval in prudence and medicine. AB - We often assume that hypothetical approval--either in the form of preferences or consent--under ideal conditions adds to the legitimacy of an arrangement or act. I want to show that this assumption, reasonable as it may seem, will also give rise to ethical problems. I focus on three problem areas: prudence, euthanasia and coercive psychiatric treatment. If we are to count as prudentially or morally relevant those preferences you would have if you were informed and rational, we will run into difficulties in all these areas if your actual and rational preferences are at variance with each other. In the prudential sphere we may question the personal value of satisfying preferences that a person does not actually have. In this case our problem concerns the point of satisfying a rational preference in conflict with an actual one. In the cases of euthanasia and coercive care it concerns instead whether it would be morally right to do such a thing. I doubt there is a simple solution to our problem. In this paper at most I prepare the way for a solution or for wiser decisions in the hard cases, by pointing out what they will have to deal with. PMID- 17080289 TI - Mucor rouxii Rho1 protein; characterization and possible role in polarized growth. AB - We have previously shown that protein kinase A of the medically important zygomycete Mucor rouxii participates in fungal morphology through cytoskeletal organization. As a first step towards finding the link between protein kinase A and cytoskeletal organization we here demonstrate the cloning of the Rho1 gene and the characterization of its protein product. The RHO1 protein primary sequence shows 70-85% identity with fungal RHO1 or mammalian RhoA. Two protein kinase A phosphorylation sequences in adequate context are predicted, Ser73 and Ser135. The peptide IRRNSQKFV, containing Ser135 proved to be a good substrate for M. rouxii protein kinase A catalytic subunit. The over-expressed Rho1 fully complements a Saccharomyces cerevisiae null mutant. The endogenous protein was identified by western blot against a developed antibody and by ADP-ribosylation. Localization in germlings was visualized by immunofluorescence; the protein was localized in patches in the mother cell surface and excluded from the germ tube. Measurement of Rho1 expression during germination indicates that Rho1, at both the mRNA and protein levels, correlates with differentiation and not with growth. Rho1 has been shown to be the regulatory protein of the beta-1,3-glucan synthase complex in fungi in which beta-1,3-glucans are major components of the cell wall. Even though glucans have not been detected in zygomycetes, caspofungin, an echinochandin known to be an inhibitor of beta-1,3-glucan synthase complex, is shown here to have a negative effect on growth and to produce an alteration on morphology when added to M. rouxii growth culture medium. This result has an important impact on the possible participation of beta-1,3-glucans on the regulation of morphology of zygomycetes. PMID- 17080288 TI - Arabidopsis immunophilins ROF1 (AtFKBP62) and ROF2 (AtFKBP65) exhibit tissue specificity, are heat-stress induced, and bind HSP90. AB - The plant co-chaperones FK506-binding proteins (FKBPs) are peptidyl prolyl cis trans isomerases that function in protein folding, signal transduction and chaperone activity. We report the characterization of the Arabidopsis large FKBPs ROF1 (AtFKBP62) and ROF2 (AtFKBP65) expression and protein accumulation patterns. Transgenic plants expressing ROF1 promoter fused to GUS reporter gene reveal that ROF1 expression is organ specific. High expression was observed in the vascular elements of roots, in hydathodes and trichomes of leaves and in stigma, sepals, and anthers. The tissue specificity and temporal expression of ROF1 and ROF2 show that they are developmentally regulated. Although ROF1 and ROF2 share 85% identity, their expression in response to heat stress is differentially regulated. Both genes are induced in plants exposed to 37 degrees C, but only ROF2 is a bonafide heat-stress protein, undetected when plants are grown at 22 degrees C. ROF1/ROF2 proteins accumulate at 37 degrees C, remain stable for at least 4 h upon recovery at 22 degrees C, whereas, their mRNA level is reduced after 1 h at 22 degrees C. By protein interaction assays, it was demonstrated, that ROF1 is a novel partner of HSP90. The five amino acids identified as essential for recognition and interaction between the mammalian chaperones and HSP90 are conserved in the plant ROF1-HSP90. We suggest that ROF/HSP90 complexes assemble in vivo. We propose that specific complexes formation between an HSP90 and ROF isoforms depends on their spatial and temporal expression. Such complexes might be regulated by environmental conditions such as heat stress or internal cues such as different hormones. PMID- 17080290 TI - Microcolonial fungi from antique marbles in Perge/Side/Termessos (Antalya/Turkey). AB - In this study rock surfaces of archaeological sites in Antalya were investigated with a focus on black fungi for the first time. Black, meristematic fungi were isolated from surfaces of antique marble monuments in Antalya (Side, Perge, Termessos). Their morphology was characterized, their diversity was documented and the taxonomy and phylogeny of new isolates was clarified based on molecular methods, that is, by sequencing parts of the small ribosomal subunit (18S) and internal transcribed spacer (ITS) regions. From a total of around 250 samples 99 different fungal strains were isolated. In most of 99 strains the rDNA sequencing data and the lack of homologies in "Genbank" gave strong evidence that these strains have to be described as new species/subspecies and/or genera. All of them, however, clustered within the ascomycete orders of Dothideales, Chaetothyriales, and Pleosporales. Field studies show that these organisms cause color changes, black spots, crater shaped lesions as well as chipping and exfoliation of the rock surfaces in Antalya and thus they have to be regarded as a serious threat to the antique cultural heritage in Turkey. PMID- 17080291 TI - 31P NMR study of polyphosphate levels during different growth phases of Phycomyces blakesleeanus. AB - The changes in relative polyphosphate content, estimated as the intensity ratio of core polyphosphate signal and intracellular inorganic phosphate signal from 31P NMR spectra, during the growth of Phycomyces blakesleeanus are reported. The ratio increases from 16 h to 28 h of growth, the minimum occurs at 32 h, followed by sharp increase up to 36 h, and a steady decrease afterwards. The changes in the biomass during mycelium growth showed steady increases, with a stagnation period between 32 h and 36 h during which a pronounced increase in the intensity ratio of core polyphosphates to intracellular inorganic phosphate signal occurred. The reduction of growth temperature from 22 degrees C to 18 degrees C significantly decreased the rate and intensity of growth, but the pattern of polyphosphate changes remained unchanged. The changes of the intensity ratio of core polyphosphates to intracellular inorganic phosphate signal are linked to characteristic stages of sporangiophore development. Analysis of core polyphosphates, intracellular inorganic phosphate and beta-ATP signal intensities suggest the role of polyphosphates as an energy and/or a phosphate reserves during Phycomyces development. PMID- 17080292 TI - Phylogenetic identification of Aeromonas strains isolated from carcasses of pig as new members of the species Aeromonas allosaccharophila. AB - The first description of the species Aeromonas allosaccharophila was only based on two strains (the type strain CECT4199, and a duplicate CECT4200) isolated from diseased elvers (Anguilla anguilla) of an eel-farm located in Valencia, Spain, and one stool isolate (ATCC35942) from a female with diarrhoea and food poisoning in South Carolina, U.S.A. In the present study, 17 Aeromonas isolates obtained from carcasses of pigs and from the equipment for the cleaning process, and one strain recently isolated from a clinical case of gastroenteritis, were genetically identified as Aerornonas allosaccharophila on the basis of gyrB and 16S rRNA gene sequencing. In addition, this phylogenetic approach also supports the classification of Aeromonas veronii biogroup sobria reference strains LMG13071, LMG13073 and LMG13074 within the species A. allosaccharophila. The A. allosaccharophila strains isolated from pig carcasses processed in a single slaughterhouse presented a clonal origin, on the basis of random amplified polymorphic DNA genetic typing. To our knowledge, this is the first time since the species description that A. allosaccharophila has been newly identified, being on this occasion isolated from the environment of a slaughterhouse. Our findings indicate that this species may be readily identified by a sequencing approach and, consequently, the present work supports the existence of this phylogenetic cluster. PMID- 17080293 TI - The role of the human DNA mismatch repair gene hMSH2 in DNA repair, cell cycle control and apoptosis: implications for pathogenesis, progression and therapy of cancer. AB - The cellular DNA mismatch repair (MMR) pathway, involving the DNA mismatch repair genes MLH1 and MSH2, detects and repairs DNA replication errors. Defects in MSH2 and MLH1 account for most cases of hereditary non-polyposis colorectal cancer as well as for sporadic colorectal tumors. Additionally, increased expression of MSH2 RNA and/or protein has been reported in various malignancies. Loss of DNA MMR in mammalian cells has been linked to resistance to certain DNA damaging agents including clinically important cytotoxic chemotherapeutics. Due to other functions besides its role in DNA repair, that include regulation of cell proliferation and apoptosis, MSH2 has recently been shown to be of importance for pathogenesis and progression of cancer. This review summarizes our present understanding of the function of MSH2 for DNA repair, cell cycle control, and apoptosis and discusses its importance for pathogenesis, progression and therapy of cancer. PMID- 17080294 TI - Langerhans cells in cutaneous tumours: immunohistochemistry study using a computer image analysis system. AB - Immunohistochemistry, based on antibody anti-S100 protein, was used to evaluate the Langerhans cells (LC) in benign and malign skin neoplasias. These cells were quantitatively estimated using a computer image analysis (OPTIMAS software system, Version 6.1) in skin biopsies diagnosed as basal cell carcinoma (BCC), epidermoid carcinoma (EpC), trichoepithelioma (TE), keratoacanthoma (KA), seborreic keratosis (SK) and actinic keratosis (AK). The antibody anti-S100 protein recognized them. No significant variations were observed in the number of LC among malignant tumour (BCC = 23.25 +/- 5.81 and EpC = 20.88 +/- 4.24). Benign lesions (AK = 33.04 +/- 7.11; TE = 55.74 +/- 9.35; SK = 42.38 +/- 9.92, and KA = 47.62 +/- 10.4) presented a higher number of LC when they were compared among them and to malignant and normal tissues. No significant differences were observed in LC area and volume between benign and malign neoplasias. These results indicate possibly differences in the immunogenicity between benign and malign epidermic tumours. In conclusion, the experimental computer assessment method was reliable and consistent to morphometric analysis of tumoural tissues. PMID- 17080295 TI - Signal assignment and secondary structure analysis of a uniformly [13C, 15N] labeled membrane protein, H +-ATP synthase subunit c, by magic-angle spinning solid-state NMR. AB - Signal assignment and secondary structural analysis of uniformly [13C, 15N] labeled H+-ATP synthase subunit c from E. coli (79 residues) in the solid state were carried out by two- and three-dimensional solid-state NMR under magic-angle spinning. The protein took on a unique structure even in the solid state from the 13C linewidths of about 1.7 ppm. On the basis of several inter- and intra-residue 13C-13C and 13C-15N chemical shift correlations, 78% of Calpha, 72% of Cbeta, 62% of C' and 61% of NH signals were assigned, which provided the secondary structure information for 84% of the 79 residues. Here, inter-residue correlations involving Gly, Ala, Pro and side-chains and a higher resolution in the 3D spectrum were significantly useful for the sequence specific assignment. On top of this, the 13C-13C correlation spectra of subunit c was analyzed by reproducing experimental cross peaks quantitatively with chemical shift prediction and signal intensity calculation based on the structure. It revealed that the subunit c in the solid state could be specified by alpha-helices with a loop structure in the middle (at sequence 41-45) as in the case of the solution structure in spite of additional extended conformations at 76-79 at the C-terminus. PMID- 17080296 TI - Genotype and allele frequency of human multidrug resistance (MDR1) gene C3435T polymorphism in Denizli province of Turkey. AB - Human p-glycoprotein encoded by human multidrug resistance (MDR1) gene, is a transmembrane protein that serves as efflux pump for a wide variety of lipophilic compounds possessing a physiological role in protecting cells against the DNA damaging of certain xenobiotics. According to the published data, the frequency of C3435T polymorphism differs depending on the different ethnical populations such as Asian, African, and Caucasians populations. In our study, we identified the MDR1 C3435T polymorphism in 150 healthy volunteers in Denizli province of Turkey. DNA was extracted from peripheral blood samples by standard phenol/chloroform extraction method. Polymerase chain reaction-restriction fragment length polymorphism was used for the detection of C3435T single nucleotide polymorphism. We obtained CC, CT and TT genotype frequencies as 20, 53 and 27%, respectively. According to our results, the C allele in Turkish population (Denizli province, west of Turkey) is found 47% and this data shows similarity with Caucasian (UK and German) populations and significantly lower than African populations (p < 0.001). Our study is the first data on the genotype and allele frequency of the human multidrug resistance (MDR1) Gene C3435T Polymorphism in Denizli Province at regional basis in Turkey. Our results could serve as a basis for large-scale correlation studies on the relevance of C3435T genotype in cancer therapy and other diseases in Turkish population. Investigation of genotype frequencies related with p-glycoprotein substrates should be investigated in large scale at regional bases in Turkish population. The scaled-up data might help either to the use of p-glycoprotein substrates to be used for therapeutic applications and population genetics considering the genotype frequencies possibly occurring throughout the history in Anatolian basin. PMID- 17080298 TI - Standard percutaneous nephrolithotomy on a pelvic kidney without any ancillary modalities. AB - We are presenting a patient with two calculi in a right pelvic kidney managed by percutaneous nephrolithotomy with an access just above the iliac crest in the prone position. PMID- 17080297 TI - Introduction of G1 phase arrest in Human Hepatocellular carcinoma cells (HHCC) by APMCF1 gene transfection through the down-regulation of TIMP3 and up-regulation of the CDK inhibitors p21. AB - We previously found that there was up-regulation of APMCF1 expression in apoptotic MCF-7 cells. Moreover, bioinformatics analysis has found that APMCF1 molecules had similar size and structure with molecules which belong to small G protein superfamily. We presume that APMCF1 plays certain biological role in the regulation of cell proliferation and apoptosis. In this study, we first detected the expression pattern of APMCF1 in human hepatocellular carcinoma cell line and find no expression in Human Hepatocellular carcinoma cells (HHCC) and enhanced expression in HepG2 cells. Expression of liposome-mediated ectogenic APMCF1 induced inhibition of HHCC growth and cell cycle, and RNAi inhibited APMCF1 expression and promoted HepG2 cell growth. Results of cell cycle gene chips analysis showed up-regulation of p21 expression and down-regulation of TIMP3 in HHCC cells expressing ectogenic APMCF1, indicating that APMCF1 participates at least partially in cell cycle regulation through regulating genes such as p21 and TIMP3. PMID- 17080299 TI - Function of interdomain alpha-helix in human brain hexokinase: covalent linkage and catalytic regulation between N- and C-terminal halves. AB - Human brain relies on a steady supply of glucose as the source of fuel, and type I hexokinase is the major isozyme governing the introduction of glucose to glycolysis in the brain. One unique regulatory property associated with type I isozyme is the alleviation of product inhibition by inorganic phosphate which binds to the N-terminal half, and the conformational change induced by inorganic phosphate must be propagated to the active site in the C-terminal half. With a single interdomain alpha-helix as the only covalent connection between the N- and C-terminal halves, the question arises as what role the interdomain alpha-helix plays at the interdomain signal transduction. Two mutants were constructed in an attempt to answer this question. The first mutant, A464P/E465G, with a helix breaker embedded in the interdomain alpha-helix had a smaller magnitude of phosphate alleviation than the wild type. The second mutant, with an insertion of seven additional residues between Gln 466 and His 467, had this phosphate relief property further diminished. Neither mutant showed dramatic changes nor the other kinetic properties. It is speculated that the interdomain alpha-helix is important for keeping the proper non-covalent contact so that transmission of the conformational changes across the N- and C-terminal half boundary can be achieved. PMID- 17080300 TI - Potential cause of lethality of an allele implicated in social evolution in fire ants. AB - The gene Gp-9 is believed to have a major effect on colony social organization in fire ants, with the presence of b-like alleles in a colony associated with multiple-queen (polygyne) organization. Queens and workers of polygyne Solenopsis invicta homozygous for the b-like allele designated b suffer reduced viability compared to other genotypes, and bb queens do not survive to become egg-layers. Thus, the b allele effectively acts as a recessive lethal. This allele differs from the remaining b-like alleles (designated b'), as well as all other Gp-9 alleles, by encoding a lysine at position 151 in the protein product, suggesting that this substitution is responsible for its deleterious effects. We tested this hypothesis by comparing frequencies of b'b' and bb homozygotes, first in queens of Solenopsis richteri and S. invicta, then in S. invicta workers from populations polymorphic for the two b-like alleles. We found that almost 20% of S. richteri queens were b'b' homozygotes, compared to the virtual absence of bb homozygotes among S. invicta queens, and that 5-18% of S. invicta workers bore genotype b'b', compared to the apparent lack of bb workers in the same populations. While we cannot entirely rule out involvement of other genes in complete gametic disequilibrium with Gp-9, our data are consistent with the hypothesis that the Lys(151) residue in GP-9 protein confers the deleterious effects of the b allele in homozygous condition, possibly by impairing the protein's function through interference with ligand binding/release or hindrance of dimer formation. PMID- 17080301 TI - Effect of surfactants on plasticizer biodegradation by Bacillus subtilis ATCC 6633. AB - The biodegradation of plasticizers has been previously shown to result in the accumulation of metabolites that are more toxic than the initial compound. The present work shows that the pattern of degradation of di-2-ethylhexyl adipate by Bacillus subtilis can be significantly altered by the presence of biosurfactants, such as surfactin, or synthetic surfactants, such as Pluronic L122. In particular, this work confirms that the monoester, mono-2-ethylhexyl adipate, is a metabolite in the breakdown of the plasticizer. This metabolite was proposed but not observed in earlier studies. Toxicity measurements showed it to be significantly more toxic than the plasticizer. Thus, the effect of the surfactants was to significantly increase the accumulation of one or both of the two most toxic metabolites; i.e., the monoester and 2-ethylhexanol. It was proposed that the most likely cause of the effect of the surfactants was the sequestering of these two metabolites into mixed micelles, resulting in their reduced availability for further degradation. PMID- 17080302 TI - Easy and rapid method of zygosity determination in transgenic mice by SYBR Green real-time quantitative PCR with a simple data analysis. AB - Establishment and maintenance of transgenic mouse strains require being able to distinguish homozygous from heterozygous animals. To date, the developed real time quantitative PCR techniques are often complicated, time-consuming and expensive. Here, we propose a very easy and rapid method with a simple data analysis to determine zygosity in transgenic mice. We show that the real-time quantitative PCR using SYBR Green fluorescent dye can be applied to discriminate two-fold differences in copy numbers of the transgene. Our procedure has to fit only three simple requirements: (1) to design primers capable of detecting one Ct difference for two-fold differences in DNA amounts (2) to measure genomic DNA concentrations accurately and (3) to have a reference animal of known zygosity in each run. Then, if the Ct values for the control gene are similar in all samples, we are able to compare directly the Ct values for the transgene in every sample, and so, to deduce the zygosity status of each mouse relative to the reference animal. This method is really simple and reliable, and it may be valuable as a rapid screening tool for zygosity status in transgenic animals. PMID- 17080303 TI - Production of endothelial nitric oxide synthase (eNOS) over-expressing piglets. AB - Vascular function, vascular structure, and homeostasis are thought to be regulated in part by nitric oxide (NO) released by endothelial cell nitric oxide synthase (eNOS), and NO released by eNOS plays an important role in modulating metabolism of skeletal and cardiac muscle in health and disease. The pig is an optimal model for human diseases because of the large number of important similarities between the genomic, metabolic and cardiovascular systems of pigs and humans. To gain a better understanding of cardiovascular regulation by eNOS we produced pigs carrying an endogenous eNOS gene driven by a Tie-2 promoter and tagged with a V5 His tag. Nuclear transfer was conducted to create these animals and the effects of two different oocyte activation treatments and two different culture systems were examined. Donor cells were electrically fused to the recipient oocytes. Electrical fusion/activation (1 mM calcium in mannitol: Treatment 1) and electrical fusion (0.1 mM calcium in mannitol)/chemical activation (200 microM Thimerosal for 10 min followed by 8 mM DTT for 30 min: Treatment 2) were used. Embryos were surgically transferred to the oviducts of gilts that exhibited estrus on the day of fusion or the day of transfer. Two cloned transgenic piglets were born from Treatment 1 and low oxygen, and another two from Treatment 2 and normal oxygen. PCR, RT-PCR, Western blotting and immunohistochemistry confirmed that the pigs were transgenic, made message, made the fusion protein and that the fusion protein localized to the endothelial cells of placental vasculature from the conceptuses as did the endogenous eNOS. Thus both activation conditions and culture systems are compatible with development to term. These pigs will serve as the founders for a colony of miniature pigs that will help to elucidate the function of eNOS in regulating muscle metabolism and the cardiorespiratory system. PMID- 17080305 TI - Sequestration of plant-derived phenolglucosides by larvae of the leaf beetle Chrysomela lapponica: thioglucosides as mechanistic probes. AB - Feeding larvae of Chrysomela lapponica (Coleoptera: Chrysomelidae) acquire characteristic O-glucosides from the leaves of their food plants. The glucosides are selectively channeled from the gut to the defensive gland. Subsequent enzymatic transformations generate a blend of different defensive compounds, e.g., salicylaldehyde and two series of 2-methylbutyl and isobutyryl esters. By using systematically modified and hydrolysis-resistant thioglucosides as structural mimics of the plant-derived glucosides, e.g., salicin and its o-, m-, and p-isomers 1, 2, and 3; o-, m-, and p-cresols 5, 6, 7; along with thioglucosides of 2-phenylethanol 9 and (3Z)-hexenol 10, we demonstrated that the larvae of C. lapponica are able to sequester a broad range of structurally different thioglucosides with comparable efficiency. This sharply contrasts with the sequestration habitus previously observed in Chrysomela populi and Phratora vitellinae, which secrete almost pure salicylaldehyde and posses a highly specific transport mechanism for salicin (Kuhn et al., Proc. Natl. Acad. Sci. USA 101:13808-13813, 2004). Also, neither C. lapponica nor C. populi sequester in their gland the thioglucoside of 8-hydroxygeraniol, the mimic of the glucoside specifically transported by larvae secreting iridoid monoterpenes (Phaedon cochleariae, Gastrophysa viridula). Accordingly, leaf beetle larvae possess selective membrane carriers in their gut and their defensive systems that match the orientation of the functional groups of glucosides from their food plants probably by embedding the substrate in a network of hydrogen bonds inside the membrane carriers. The synthesis and the spectroscopic properties of the test compounds along with a comparative evaluation of the transport capabilities of larvae of C. populi and C. lapponica are described. PMID- 17080304 TI - Glucosylceramide transfer from lysosomes--the missing link in molecular pathology of glucosylceramidase deficiency: a hypothesis based on existing data. AB - Gaucher disease (GD), deficiency of acid glucosylceramidase (GlcCer-ase) is characterized by deficient degradation of beta-glucosylceramide (GlcCer). It is well known that, in GD, the lysosomal accumulation of uncleaved GlcCer is limited to macrophages, which are gradually converted to storage cells with well known cytology--Gaucher cells (GCs). On the basis of previous studies of the disorder and of a comparison with other lysosomal enzymopathies affecting degradation of the GlcCer-based glycosphingolipid series, it is hypothesized that in other cell types (i.e. non-macrophage cells) the uncleaved GlcCer, in GlcCer-ase deficiency, is transferred to other cell compartments, where it may be processed and even accumulated to various degrees. The consequence of the abnormal extralysosomal load may differ according to the cell type and compartment targeted and may be influenced by genetically determined factors, by a number of acquired conditions, including the current metabolic situation. The sequelae of the uncleaved GlcCer extralysosomal transfer may range from probably innocent or positive stimulatory, to the much more serious, in which it interferes with a variety of cell functions, and in extreme cases, can lead to cell death. This alternative processing of uncleaved GlcCer may help to explain tissue alterations seen in GD that have, so far, resisted explanation based simply on the presence of GCs. Paralysosomal alternative processing may thus go a long way towards filling a long-standing gap in the understanding of the molecular pathology of the disorder. The impact of this alternative process will most likely be inversely proportional to the level of residual GlcCer-ase activity. Lysosomal sequestration of GlcCer in these cells is either absent or in those exceptional cases where it does occur, it is exceptional and rudimentary. It is suggested that paralysosomal alternative processing of uncleaved GlcCer is the main target for enzyme replacement therapy. The mechanism responsible for GlcCer transfer remains to be elucidated. It may also help in explaining the so far unclear origin of glucosylsphingosine (GlcSph) and define the mutual relation between these two processes. PMID- 17080306 TI - Electrophysiological responses of the lepidopterous stemborers Chilo partellus and Busseola fusca to volatiles from wild and cultivated host plants. AB - The stemborers Chilo partellus and Busseola fusca are major pests of subsistence cereal farming in Africa. Volatiles released by two cultivated hosts, sorghum and maize (Sorghum bicolor and Zea mays), and two wild grass hosts, Pennisetum purpureum and Hyparrhenia tamba, were collected by air entrainment. Electrophysiologically active components in these samples were detected by coupled gas chromatography-electroantennography (GC-EAG), and the active peaks identified by gas chromatography-mass spectrometry. A total of 41 compounds were identified from the four plant species, all of which, as well as two unidentified compounds, elicited an electrophysiological response from one or both of the stemborers. The compounds included a number of green leaf volatiles and other aliphatic aldehydes, ketones, and esters, mono- and sesquiterpenoids, and some aromatic compounds. EAG studies with authentic samples, conducted at two discriminating doses for all compounds, and dose-response curves for 14 of the most highly EAG-active compounds, showed significant differences in relative responses between species. The compounds that elicited large responses in both species of moths included linalool, acetophenone, and 4-allylanisole, while a number of compounds such as the aliphatic aldehydes octanal, nonanal, and decanal elicited a large response in B. fusca, but a significantly smaller response in C. partellus. Furthermore, the wild hosts produced higher levels of physiologically active compounds compared with either of the cultivated hosts. These differences are discussed in relation to the differential attraction/oviposition of the two stemborers observed in the field and, particularly for eastern African small scale farming systems, in the context of using a push-pull strategy for their control. PMID- 17080307 TI - Metabolomic differentiation of Brassica rapa following herbivory by different insect instars using two-dimensional nuclear magnetic resonance spectroscopy. AB - The metabolic alterations of Brassica rapa (L.) leaves attacked by larvae of the specialist Plutella xylostella L. (Lepidoptera: Yponomeutidae) and the generalist Spodoptera exigua Hubner (Lepidoptera: Noctuidae) were investigated with nuclear magnetic resonance (NMR) spectroscopy, followed by a multivariate data analysis. The principal component analysis (PCA) of (1)H NMR spectra showed that metabolic changes in B. rapa leaves induced by the 2nd and the 4th instars were different from each other. However, the congestion of the one-dimensional (1)H NMR spectrum made it difficult to identify discriminating metabolites. To overcome the spectral complexity, several two-dimensional NMR techniques were applied. Of those evaluated, J-resolved spectroscopy, which affords an additional coupling constant, provided a wide range of structure information on differentiating the metabolites. Based on the J-resolved spectra combined with PCA, the major signals contributing to the discrimination were alanine, threonine, glucose, sucrose, feruloyl malate, sinapoyl malate, and gluconapin. PMID- 17080308 TI - MDM2 SNP309 accelerates familial breast carcinogenesis independently of estrogen signaling. AB - A single nucleotide polymorphism (SNP309T>G) in the intronic promoter of MDM2 was recently found to accelerate carcinogenesis in early-onset cancer cases. This cancer acceleration presumably was due to increased SP1 binding, resulting in enhanced MDM2 transcriptional activation by estrogens. We evaluated MDM2 SNP309 in 343 familial breast cancer cases with known mutation status for CHEK2 1100delC, BRCA1 and BRCA2. Cancer acceleration was indeed observed in early-onset familial breast cancer cases (diagnosed or = 25 kg between 15 and age 50 was inversely associated with breast cancer among Hispanic women (OR 0.51, 95% CI 0.23, 1.14). A large weight gain and a large waist-to-hip ratio (WHR) was associated with an increased odds of having an estrogen receptor negative tumor among NHW only (OR 1.81, 95% CI 1.07, 3.08, and 2.04 95% CI 1.20,3.50). CONCLUSIONS: These findings suggest that the metabolic consequences of obesity on breast cancer risk differ between NHW and Hispanic women living in the Southwest. PMID- 17080311 TI - Release of secondary free radicals during post-ischaemic reperfusion is not influenced by extracellular calcium levels in isolated rat hearts. AB - In this study, we evaluated the impact of the calcium concentration present in the perfusion medium (1.2-3 mM) on contractile performance, lactate dehydrogenase (LDH) release and secondary free radical production during post-ischaemic reperfusion of isolated rat hearts. The impact of calcium concentration on post ischaemic free radical release was investigated using the Electron Paramagnetic Resonance (EPR) technique and spin trapping with the lipophilic spin trap alpha phenyl N-tert-butylnitrone (PBN). The evolution of left ventricular end diastolic pressure (LVEDP) in both groups followed the same pattern, but we observed that ischaemic and post-ischaemic contracture was more severe in the group of hearts perfused with 3 mM of calcium as compared with those perfused with 1.2 mM of calcium. A large release of alkyl/alkoxyl species occurred in all hearts from the onset of reperfusion and remained at a high level during the 30 min of reperfusion with no return to basal values. The kinetics and intensity of these releases were the same in both groups. In conclusion, in a range of extracellular calcium levels (1.2-3 mM), the release of alkyl/alkoxyls radicals does not seem to be calcium-dependent. Due to the protective actions of PBN itself, the results of simultaneous investigations of the effects of radical scavengers on isolated heart function may be limited. However, since many pharmacological properties (antioxidant, cellular protector, NO precursor ...) are attributed to PBN, studies investigating oxidative stress with such a multi-faceted tool make interpretation difficult. PMID- 17080314 TI - Endocrine disruption in male mosquitofish (Gambusia holbrooki) inhabiting wetlands in Western Australia. AB - The use of gonopodial indices as potential indicators of endocrine disruption in the mosquitofish Gambusia holbrooki inhabiting south west Australian wetlands was investigated. A minimum of 50 mature males was collected from each of five water bodies in the Swan Coastal Plain, Western Australia, in order to measure morphological features related to reproduction. A set of morphological measurements were used to derive the following indices: gonopodium length/standard body length, pre-anal length/standard body length, the index of elongation and the percentage of male fish with hooks on the distal end of the gonopodium. Indices of male mosquitofish collected from Jack Finney Lake, located in the Curtin University campus, suggest the presence of endocrine disrupting chemicals (EDCs) in this water-body, while those from Lake Kulinup suggest this is a site of concern. Indices of male fish from the Wagerup wetland, Lake Monger and Loch McNess indicate that fish inhabiting these wetlands are not affected by EDCs. This preliminary study suggests that EDCs may be present in a number of wetlands of the Swan Coastal Plain. Further study using EDC specific markers such as vitellogenin induction in male mosquitofish is required to confirm whether EDCs are present in these water-bodies. PMID- 17080313 TI - The CNS synapse revisited: gaps, adhesive welds, and borders. AB - Although processes leading up to the point of synapse formation are fairly well understood, the precise sequence of events in which the membranes of two separate cells "lock in" to form a mature synaptic junctional complex is poorly understood. A careful study of the molecules operating at the synapse indicates that their roles are more multifarious than once imagined. In this review we posit that the synapse is a functional organelle with poorly defined boundaries and a complex biochemistry. The role of adhesion molecules, including the integration of their signaling and adhesive properties in the context of synaptic activity is discussed. PMID- 17080315 TI - Understanding effects of matrix protease and matrix organization on directional persistence and translational speed in three-dimensional cell migration. AB - Recent studies have shown significant differences in migration mechanisms between two- and three-dimensional environments. While experiments have suggested a strong dependence of in vivo migration on both structure and proteolytic activity, the underlying biophysics of such dependence has not been studied adequately. In addition, the existing models of persistent random walk migration are primarily based on two-dimensional movement and do not account for the effect of proteolysis or matrix inhomogeneity. Using lattice Monte Carlo methods, we present a model to study the role of matrix metallo-proteases (MMPs) on directional persistence and speed. The simulations account for a given cell's ability to deform as well as to digest the matrix as the cell moves in three dimensions. Our results show a bimodal dependence of speed and persistence on matrix pore size and suggest high sensitivity on MMP activity, which is in very good agreement with experimental studies carried out in 3D matrices. PMID- 17080316 TI - Circumstances of pregnancy: low income women in Georgia describe the difference between planned and unplanned pregnancies. AB - INTRODUCTION AND BACKGROUND: The United States has a higher rate of unintended pregnancies than many other developed countries despite the availability of effective contraception. Using Azjen's Theory of Planned Behavior and Luker's Theory of Contraceptive Risk Taking this qualitative study explores the reasons for the high rate of unintended pregnancy among low income women in Georgia. METHODS: Six focus groups (N=39) were conducted at county health department clinics. Women were recruited at the clinics to participate in a discussion of Pregnancy and Motherhood. All English or Spanish speaking women between the ages of 18 and 49 were eligible for participation. RESULTS: Three dimensions of pregnancy planning emerged-the decision to have a child, taking specific behaviors to increase the likelihood of conception, and making plans to care for a child. Planned pregnancies may occur when a couple or a woman decides that they are ready to have a child, but also occur in less stable situations that pose risks for the family and the child. Unplanned pregnancies may be welcomed or be unwanted depending on the circumstances. Women perceive advantages and disadvantages to planned and unplanned pregnancies. DISCUSSION: There are major differences between the definition and values of planned and unplanned pregnancies promoted by public health practitioners and those of the women in our study. This suggests the need for research on the circumstances in which pregnancies occur, as well as targeted interventions and policies that help women and men define and achieve the circumstances in which they would like to bear and rear children. PMID- 17080317 TI - Leukocytes in Complex Regional Pain Syndrome type I. AB - OBJECTIVE: The pathophysiology of Complex Regional Pain Syndrome type I (CRPS I) is unclear. An inflammatory reaction may cause the syndrome in which leukocytes may play an important role. MATERIALS AND METHODS: In this pilot study of six patients with acute warm CRPS I, we performed radiolabeled autologous leukocyte scans of both hands, in order to assess leukocyte accumulation. Comparison was made with the unaffected limb, and with three control patients with a Colles fracture without CRPS I. RESULTS: Images of the CRPS I patients obtained 4 h after leukocyte injection provided the clearest results. At 4 h post-injection, there was clear, asymmetrical leukocyte accumulation in the affected extremity with a mean ratio of 1.49+/-0.19. In control patients, no asymmetry was observed between hands (mean ratio 1.09+/-0.06), indicating the absence of specific leukocyte accumulation. There was a statistically significant difference between CRPS I and control subjects 4 h post injection (p=0.012). CONCLUSION: We found a significantly increased accumulation of leukocytes in patients with CRPS I. This is the first study to show a possible role for leukocytes in the pathophysiology of acute CRPS I. PMID- 17080318 TI - Phonological activation in multi-syllabic sord recognition. AB - Three experiments were conducted to test the phonological recoding hypothesis in visual word recognition. Most studies on this issue have been conducted using mono-syllabic words, eventually constructing various models of phonological processing. Yet in many languages including English, the majority of words are multisyllabic words. English includes words incorporating a silent letter in their letter strings (e.g., champagne). Such words provide an opportunity for investigating the role of phonological information in multi-syllabic words by comparing them to words that do not have the silent letter in the corresponding position (e.g., passenger). The performance focus is on the effects of removing letters from words with a silent letter and from words with a non-silent letter. Three representative lexical tasks--naming, semantic categorization, lexical decision--were conducted in the present study. Stimuli that excluded a silent letter (e.g., champa_ne) were processed faster than those that excluded a sounding letter (e.g., passen_er) in the naming (Experiment 1), the semantic categorization (Experiment 2), and the lexical decision task (Experiment 3). The convergent evidence from these three experiments provides seminal proof of phonological recoding in multi-syllabic word recognition. PMID- 17080320 TI - Bipolar disorders in Australia. A population-based study of excess costs. AB - OBJECTIVE: To estimate the excess costs associated with bipolar disorders in Australia, based on prevalence (using the Mood Disorder Questionnaire (MDQ)) and associated excess burden-of-illness costs. METHODS: Using data from the 2004 South Australian Health Omnibus Survey (HOS), a weighted cross-sectional survey of 3,015 adults, excess costs were estimated from health service utilisation. RESULTS: There was a 2.5% lifetime prevalence of bipolar disorders, delineated by the MDQ. Those persons (MDQ positive) reported a significantly greater use of services and a poorer health status and quality of life than those who were MDQ negative. Using the service provision perspective, excess costs of bipolar disorders in Australia were approx $3.97-$4.95 billion. CONCLUSIONS: These results from an Australian population demonstrate the significant economic burden of bipolar disorders. Our findings emphasise the need for further evaluation of the cost-effectiveness of different treatments, or alternative means of reducing the burden borne by individuals, the health system and the general community. PMID- 17080321 TI - Short-term outcome of inpatient psychiatric care--impact of coercion and treatment characteristics. AB - AIMS: Little is known about the outcome of brief inpatient treatment interventions in routine psychiatric practice. The aim of this article was to study if subjective and assessed outcome of brief psychiatric inpatient care are related to patient characteristics, coercion at admission and during care, and other treatment characteristics. METHOD: A total of 233 involuntarily and voluntarily admitted patients were interviewed within 5 days from admission and at discharge or after 3 weeks of care. Outcome was measured as reported by patients and by change in GAF (Global Assessment Scale) scores. RESULTS: Predictors for a positive subjective outcome were if the patients reported that they had been well treated by the staff and had contact persons at the ward. Predictors for a GAF improvement were a low GAF score at admission and a mood disorder diagnosis. CONCLUSIONS: Subjectively reported outcome and outcome measured by assessing change in level of functioning differed. Coercion was not related to outcome. The way the patient perceived they had been treated by the staff was strongly related to subjective outcome. PMID- 17080322 TI - Mental health in three generations of Iranian medical students and doctors. A cross-sectional study. AB - OBJECTIVE: Despite research in Western countries finding high levels of psychological morbidity in medical students and doctors, little is known about difficulties faced by medical staff in the Middle East. The aim of the present study was to assess emotional disturbance and interpersonal attitudes in a representative sample of Iranian medical students and practitioners. METHOD: A total of 82 medical students, 92 interns and 89 general practitioners (GPs) participated in the study. Participants rated the 28-item version of the General Health Questionnaire (GHQ-28) and a measure of interpersonal attitudes. RESULTS: A total of 44% of participants scored above the threshold of the GHQ-28, indicating probable psychiatric disorder. The GHQ-28 scores were higher in students than interns or GPs and in women compared with men. Medical students and doctors had high levels of indifference and cynicism. CONCLUSIONS: This study suggests that psychological morbidity was common in Iranian medical students and practitioners, particularly women. Women were at particular risk. A high prevalence of emotional disturbance among health care practitioners is likely to compound existing problems of health care provision. PMID- 17080323 TI - Ethnic disparities in problem behaviour in adolescence contribute to ethnic disparities in social class in adulthood. AB - BACKGROUND: It is important for prevention of social class disparities to know how ethnic disparities in social class arise among migrant children. We contribute to this understanding by examining the role of problem behaviour in adolescence. METHODS: Prospective observational study with 753 Dutch native and 217 Turkish migrant adolescents (11-18 year) followed for 10 years. Internalising and externalising problems were assessed in adolescence and employment status and occupational level were assessed in adulthood. The difference in odds ratios (OR) before and after adjustment for internalising and externalising problems was an indication of the predictive value of disparities in internalising and externalising problems for the development of social class disparities. RESULTS: A total of 135 (62%) of the Turkish and 602 (80%) of the Dutch adults were employed. Internalising and externalising problems were not associated with employment status. Of the employed, 65 (48%) Turkish and 179 (30%) Dutch adults worked in low-level occupations (p < 0.0001). Internalising and externalising problems were associated with both ethnicity and occupation. The OR for low-level occupation for Turkish adults was 1.78 (1.19-2.65), indicating ethnic disparities. Adjustment for internalising problems lowered the OR with 36% to 1.50 (0.97-2.31), and adjustment for externalising problems lowered it with 8% to 1.72 (1.15-2.57). Findings were similar for men and women and did not vary by age. CONCLUSIONS: Ethnic disparities in occupational level in adulthood could partly be attributed to disparities in mental health between Turkish migrants and Dutch natives in adolescence. Prevention of ethnic disparities in mental health at young age may therefore also contribute to the prevention of occupational differences in adulthood. PMID- 17080324 TI - Multiple exposure to life events and clinical psychopathology in adults with intellectual disability. AB - BACKGROUND: Relatively few studies have examined the impact of life events on mental health in the field of intellectual disability (ID), despite the possibility that adults with ID and mental health problems might be particularly vulnerable to multiple events. AIMS: To examine the impact of multiple life events on mental health in people with ID. METHOD: The sample consisted of 281 men and women with ID consecutively referred to a specialist mental health service in South-East London. For each participant, key informants completed the PAS-ADD Life Events list, while psychiatric diagnoses were made by clinicians based on ICD-10 criteria. RESULTS: Logistic regression analysis revealed that single exposure to life events was significantly associated with female gender, schizophrenia, personality disorders and depression. However, multiple exposure to life events was associated with personality disorder, depression and adjustment reaction. CONCLUSIONS: The present results suggest an increased vulnerability to life events in people with ID. A better understanding of the relationship between life events and mental health can enhance the development of interventions, which may benefit substantially this particular population. PMID- 17080325 TI - Sesquiterpene lactones induce distinct forms of cell death that modulate human monocyte-derived macrophage responses. AB - Sesquiterpene lactones (SQTLs) are shown to possess anti-inflammatory as well as cytotoxic activity. No study, however, links both activities. We, therefore, hypothesized that SQTL-treated, dying cells might induce an anti-inflammatory response in cocultured THP-1 macrophages. Here we show that SQTLs bearing either an alpha,beta-unsaturated cyclopentenone or an alpha-methylene-gamma-lactone induce different forms of cell death. Whereas the cyclopentenone SQTL induced typical apoptosis, the alpha-methylene-gamma-lactone SQTLs-induced cell death lacked partly classical signs of apoptosis, such as DNA fragmentation. All SQTLs, however, activated caspases and the nuclear morphology of cell death was dependent on caspase activation. Most interestingly, alpha-methylene-gamma lactone SQTLs induced a more pronounced phosphatidylserine (PS) exposure than the cyclopentenone SQTL. Especially, 7-hydroxycostunolide (HC), with an alpha methylene-gamma-lactone substituted with a hydroxyl group, showed a striking fast and pronounced PS translocation. This result was in agreement with a strong activation of phagocytosis in cocultured THP-1 macrophages. Interestingly, HC treated Jurkat cells led to an early (3.5 h) but transient increase in TNF-alpha levels in macrophage coculture. Release of TGF-beta remained unaffected after 18 h. We propose that this type of SQTL may influence local inflammation by transiently activating the immune system and help to clear cells by inducing a form of cell death that promotes phagocytosis. PMID- 17080326 TI - Herpes simplex virus blocks apoptosis by precluding mitochondrial cytochrome c release independent of caspase activation in infected human epithelial cells. AB - Expression of HSV-1 genes leads to the induction of apoptosis in human epithelial HEp-2 cells but the subsequent synthesis of infected cell protein prevents the process from killing the cells. Thus, viruses unable to produce appropriate prevention factors are apoptotic. We now report that the addition of either a pancaspase inhibitor or caspase-9-specific inhibitor prevented cells infected with an apoptotic HSV-1 virus from undergoing cell death. This result indicated that HSV-1-dependent apoptosis proceeds through the mitochondrial apoptotic pathway. However, the pancaspase inhibitor did not prevent the release of cytochrome c from mitochondria, implying that caspase activation is not required for this induction of cytochrome c release by HSV-1. The release of cytochrome c was first detected at 9 hpi while caspase-9, caspase-3 and PARP processing were detected at 12 hpi. Finally, Bax accumulated at mitochondria during apoptotic, but not wild type HSV-1 infection. Together, these findings indicate that HSV-1 blocks apoptosis by precluding mitochondrial cytochrome c release in a caspase independent manner and suggest Bax as a target in infected human epithelial cells. PMID- 17080327 TI - Fusiogenic endogenous-retroviral syncytin-1 exerts anti-apoptotic functions in staurosporine-challenged CHO cells. AB - Fusiogenic glycoprotein syncytin-1, expressed in human placenta, is a promising candidate for acquiring a basic knowledge of placental syncytialization. However, its cellular mode of action is unidentified. We investigated whether syncytin-1 may exert influence on apoptotic processes. Therefore, we incubated CHO cells after stable transfection with syncytin-1 (CHO-52) in the presence or absence of staurosporine (STS), a kinase inhibitor well characterized to induce apoptosis. When testing the phenotype of CHO-52 cells, we could demonstrate that the induction of apoptosis by STS was delayed over a period of up to 24 h. Furthermore, the cell death rate was decreased by approx 75% following transfection of syncytin-1 in CHO-52 compared to mock-treated cells. In detail, after 18h of incubation with 500 nM STS, 64 +/- 2% of CHO-52 cells were viable compared to 16 +/- 1% of CHO-mocks, after 24 h 43 +/- 3% vs 5 +/- 2%, respectively. CHO-52 cells exhibited a lower expression of active caspase 3 and anti-apoptotic Bcl-2 was found to be increased in CHO-52 cells at baseline and following STS treatment. Our study provides first evidence that syncytin-1 serves anti-apoptotic function under certain conditions. A lessened activation of caspase 3 and an increased expression of Bcl-2 are possible mechanisms. PMID- 17080328 TI - Mitochondrial control of cell death induced by hyperosmotic stress. AB - HeLa and HCT116 cells respond differentially to sorbitol, an osmolyte able to induce hypertonic stress. In these models, sorbitol promoted the phenotypic manifestations of early apoptosis followed by complete loss of viability in a time-, dose-, and cell type-specific fashion, by eliciting distinct yet partially overlapping molecular pathways. In HCT116 but not in HeLa cells, sorbitol caused the mitochondrial release of the caspase-independent death effector AIF, whereas in both cell lines cytochrome c was retained in mitochondria. Despite cytochrome c retention, HeLa cells exhibited the progressive activation of caspase-3, presumably due to the prior activation of caspase-8. Accordingly, caspase inhibition prevented sorbitol-induced killing in HeLa, but only partially in HCT116 cells. Both the knock-out of Bax in HCT116 cells and the knock-down of Bax in A549 cells by RNA interference reduced the AIF release and/or the mitochondrial alterations. While the knock-down of Bcl-2/Bcl-X(L) sensitized to sorbitol-induced killing, overexpression of a Bcl-2 variant that specifically localizes to mitochondria (but not of the wild-type nor of a endoplasmic reticulum-targeted form) strongly inhibited sorbitol effects. Thus, hyperosmotic stress kills cells by triggering different molecular pathways, which converge at mitochondria where pro- and anti-apoptotic members of the Bcl-2 family exert their control. PMID- 17080329 TI - [Measures of quality assurance for in-patient pediatric oncology units]. PMID- 17080330 TI - Pathology and molecular biology of teratomas in childhood and adolescence. AB - The biologic behaviour of teratomas depends on various interdependent clinical and epidemiologic variables such as the age at diagnosis, sex, tumor site, histology which all correlate to different cytogenetic and molecular biologic aberrations. Thus, testicular teratomas of infancy are generally benign. Accordingly, prepubertal teratomas show no cytogenetic or molecular genetic aberrations. In contrast, postpubertal testicular teratomas can present as clinically malignant tumors and may show complex cytogenetic aberrations such as the isochromosome 12p, which is pathognomonic of malignant germ cell tumors. Notably, teratomas of both age groups show an at least partial erasure of the genomic imprinting, correlating with their origin from primordial germ cells. The Kiel Pediatric Tumor Registry includes 541 teratoma specimens, and among these, the most frequent tumor sites (in descending order) are: the sacrococcygeal region (33.8 %), the ovaries (31.2 %) and the testes (10.5 %). Rare localizations include the mediastinum, the retroperitoneum, the head and neck region as well as the central nervous system. The WHO classification of germ cell tumors distinguishes mature and immature teratomas as well as teratomas with malignant transformation. In immature teratomas, primitive neuroectodermal structures predominate. According to the grading system (Gonzalez-Crussi, 1982), mature teratomas (G0) are more frequent (54.5 %) than immature teratomas (G1-G3, 45.5 %). Only 7.8 % of all teratomas show the highest grade of immaturity (G3). The frequency of additional microscopic foci of malignant yolk sac tumor correlates with the grade of immaturity. In sacrococcygeal teratomas, the yolk sac tumor microfoci may give rise to a malignant relapse after incomplete resection. The rare teratomas with malignant transformation contain components with "conventional" somatic type malignancy such as leukaemia, carcinoma or sarcoma. Here, molecular genetic analysis has demonstrated the origin of the somatic malignancy from a malignant transformation within the germ cell tumor with retention of the cytogenetic changes characteristic of malignant germ cell tumors. PMID- 17080331 TI - Teratoma with malignant transformation: experiences of the cooperative GPOH protocols MAKEI 83/86/89/96. AB - BACKGROUND: The designation of a teratoma with malignant transformation (TMT) refers to the occurrence of somatic non-germ cell malignancies within a teratoma. While TMT is a rare but well recognised phenomenon in adult germ cell tumors (GCT), data on TMT in pediatric GCTs are lacking. PATIENTS AND METHODS: Between 1982 and 2003, 641 patients with extracranial nontesticular pure teratoma (256 coccygeal, 246 ovarian, 139 other sites) were reported to the MAKEI protocols 83/86/89/96 by various, mainly German centres. Patients with teratoma and somatic malignancy were identified by database queries. Patients with malignant germ cell tumor components were excluded from this analysis. The clinical files were reviewed and re-evaluated centrally. Information was actualized by requisition to the participating hospitals. RESULTS: We identified nine patients (all female, age 0-39 years) meeting the histological criteria of TMT. Two patients had coccygeal teratomas and seven ovarian tumors. Carcinoma was the dominating malignancy (five of nine cases). Tumors with glial differentiation and embryonal tumors occurred in two cases each. Resection was performed in seven patients (including both coccygeal tumors) and adjuvant chemotherapy was administered in one of them. Two patients relapsed after resection, but both were cured with chemotherapy. Two patients suffered from advanced tumors and both were treated with primary chemotherapy. One patient was cured from the malignant component (astrocytoma), but the teratomatous components persisted. The other patient died as a result of progression of her malignant medulloepithelioma. CONCLUSIONS: Malignant transformation of pure teratomas constitutes a very rare entity in children and adolescents that is most commonly observed in postpubertal girls with ovarian teratoma. Compared to adult patients, similar malignant entities can be observed in association with teratoma. However, in our series, no sarcoma was diagnosed. In localised tumors, complete resection appears to be adequate, whereas chemotherapy should be considered in patients with R1- or R2-resection. Cisplatinum-based chemotherapy was effective as two of four relapsed patients survived tumor free. However, the ideal regimen has not yet been established and the known sensitivity of the histologic components to cytostatic drugs has to be considered in the choice of treatment. Further molecular biologic studies are necessary to understand the origin of these tumors. PMID- 17080332 TI - The malignant potential of teratomas in infancy and childhood: the MAKEI experiences in non-testicular teratoma and implications for a new protocol. AB - Since 1982, mature and immature teratomas have been recruited into the MAHO and MAKEI protocols of the German Society for Pediatric Oncology and Hematology (GPOH) for testicular and non-testicular germ cell tumors in order to study the epidemiology and clinical behaviour of teratomas. Patients were registered in the epidemiologic German Childrens Cancer Registry and the GPOH Childrens Tumor Registry for pathological review. Patients with immaturity grade 2 and 3 according to Gonzales-Crussi were eligible for adjuvant chemotherapy. The consecutive protocols MAKEI 83/86/89 have been published previously in detail (Klin Paediatr 1997; 209: 228-234, Med Pediat Oncol 1998; 31: 8-15) and will be compared to the data of MAKEI 96. For this comparison, 274 patients from MAKEI 83/86/89 and 261 patients from MAKEI 96 are evaluable. RESULTS: 1) EFS after complete tumor resection has been estimated to 0.96 +/- 0.01 in both observation periods. 2) Incomplete tumor resection remains the main risk factor for relapse (EFS 0.55 +/- 0.09). 3) The relapse rate declined from 13.9 % in MAKEI 83/86/89 to 9.5 % in MAKEI 96. 4) In MAKEI 83/86/89 four newborns with teratoma died due to perioperative complications and nine children as a result of tumor progression, whereas in MAKEI 96 no newborn died, only one child died from tumor progression, and another child died during long time observation for another reason (meningitis). 5) In accordance to the experience of the MAKEI 83/86/89 studies, no child of the MAKEI 96 study presented with yolk sac tumor at recurrence if adjuvant chemotherapy was administered during first-line treatment because of immaturity. In contrast, more than half of the children with tumor recurrence after watch and wait strategy had yolk sac tumor in addition to teratoma. PMID- 17080333 TI - Relapse pattern after complete resection and early progression after incomplete resection of childhood craniopharyngioma. AB - In HIT Endo data on therapy and prognosis of 306 patients with childhood craniopharyngioma (CP) were analyzed. The 5 years-overall survival rate was 94 +/ 4 % in irradiated patients and 93 +/- 5 % in non-irradiated patients. Aims of the prospective study KRANIOPHARYNGEOM 2000 were to collect data on the incidence and time course of relapses after complete surgery and tumour progressions after incomplete resection. Furthermore, the impact of irradiation therapy (XRT) on tumour relapse and recurrence rates was analyzed. Since 2001 ninety-eight patients with CP were recruited at a median age at diagnosis of 9.9 years ranging from 1.8 to 18.0 years. Complete resection was achieved in 44 %, incomplete resection in 54 %. XRT was performed in 24 of 98 CP patients; in 10 early after incomplete resection, in 14 of 24 after progression of residual tumour or relapse, in 3 of 14 after second surgery of relapse. XRT was performed at a median age of 12.0 years ranging from 5.0 to 18.9 years and in median after an interval of 9 months after first diagnosis. The analysis of event-free survival rates (EFS) in patients with CP showed a high rate of early events in terms of tumour progression after incomplete resection (3y-EFS: 0.22 +/- 0.09) and relapses after complete resection (3y-EFS: 0.60 +/- 0.10) during the first three years of follow-up. A high rate of early events (1y-EFS: 0.78 +/- 0.10; 2y-EFS: 0.57 +/- 0.15) was also found for patients after XRT (3 cystic progressions, 3 progressions of solid tumour; in 24 patients after XRT). We conclude that tumour progression and relapse are frequent and early events even in irradiated patients. Monitoring of cerebral imaging and clinical status is recommended in follow-up of patients with childhood CP. In order to analyze the appropriate time point of XRT after incomplete resection, QoL, EFS and overall survival in patients (age > or = 5 years) will be analyzed in KRANIOPHARYNGEOM 2007 after stratified randomization of the time point of irradiation after incomplete resection (early irradiation versus irradiation at progression of residual tumour). PMID- 17080334 TI - Haploidentical stem cell transplantation in patients with pediatric solid tumors: preliminary results of a pilot study and analysis of graft versus tumor effects. AB - Pediatric patients with relapsed metastatic tumors have a poor prognosis and new treatment strategies are warranted. We present preliminary results of a pilot study, evaluating the feasibility and toxicity of transplantation of haploidentical T and B cell depleted grafts with high numbers of NK cells. 6 patients with relapsed metastatic neuroblastomas (n = 4), rhabdomyosarcoma (n = 1) or Ewing's sarcoma (n = 1) after previous autologous transplantation received CD3/CD19 depleted grafts from mismatched family donors with a median number of 16 x 10 (6)/kg stem cells, 167 x 10 (6)/kg Natural Killer cells and only 5.4 x 10 (4)/kg residual T cells. A melphalan-based, reduced intensity conditioning was used. Despite pretransplant chemotherapy, patients entered transplantation with significant tumor burden. Primary engraftment occurred in 6/6 patients. One patient had secondary graft failure. Hematopoietic recovery was rapid (ANC > 0.5 x 10 (9)/L: 11 days (9-12); independence from platelet substitution: 8 days (7 11)). Four patients had acute GvHD grade II, limited chronic GvHD was observed in 2 patients. No transplant-related mortality and only low toxicity occurred. Four patients died from progression, two patients are alive. Overall median survival time is 6 months (2-11) to date. Analysis of posttransplant NK cell function revealed stable cytotoxic activity against K562 targets, whereas activity against neuroblastoma targets was low. Stimulation with cytokines and use of appropriate antibodies clearly enhanced specific lysis in vitro. In summary, these preliminary results indicate the feasibility and low toxicity even in intensively pre-treated patients with neuroblastomas/sarcomas. This approach may form the basis for posttransplant immunomodulation and other therapeutic strategies. Further experience is warranted to evaluate the method. PMID- 17080335 TI - Identification of candidate target antigens for antibody-based immunotherapy in childhood B-cell precursor ALL. AB - BACKGROUND: Contemporary risk adapted treatment protocols for childhood acute lymphoblastic leukemia (ALL) rely on accurate risk assessment strategies for disease re-occurrence by incorporating clinical parameters as well as immunological, molecular and cytogenetic features of the blasts at initial manifestation. Additional risk stratification is provided by analysis of the IN VITRO and IN VIVO response of the blasts towards standard chemotherapy. Despite adapted therapies, a number of children with good and bad prognostic factors still fail therapy. One approach to this problem might be to incorporate monoclonal antibodies (MoAbs) as additional modalities into the first or second line treatment. PATIENTS AND METHODS: In order to identify target antigen structures, we analyzed the immunological expression profiles of blasts from 181 patients with B-cell precursor ALL treated at our institution in 11 years according to the CoALL-92/97/03 protocols. Blasts were classified according to the EGIL guidelines as 9 proB-, 110 common (c-) and 62 preB-ALL. RESULTS: > 99 and 96 % of patients expressed CD19 and CD22 on > 90 % of their blasts, respectively. HLA-DR on > 95 % blasts was present in all patients. CD10 was expressed on all c-/preB-ALL and absent on proB-ALL cells. CD20 was expressed on 11-37 % of B-cell precursor ALL samples. CD34 positive blasts were found in 89, 83 and 68 % of patients with proB-, c- and preB-ALL, respectively. CD37 expression was detected in 0-18 % of patients. < 20 % CD45(+) blasts were found in 11, 19 and 18 % of patients with proB-, c- and preB-ALL. CD33(+) was expressed on 33, 29 and 21 % of patients samples with proB-, c- and preB-ALL. Other myeloid antigens (CD13, CD14, CD15, CD65) were positive on blasts in < 25 % of patients. Analyses of the immunological profile of blasts in 9 consecutive children with relapse revealed that the antigen expression profile varied little compared to the initial diagnosis for CD10, CD19, CD22 and HLA-DR. CONCLUSIONS: These analyses clearly identified the three antigens CD19, CD22 and HLA-DR present on blasts in more than 90 % of patients as potential target structures for targeted therapies with native or toxin-bound monoclonal antibodies in childhood ALL. PMID- 17080336 TI - Preoperative screening for coagulation disorders in children undergoing adenoidectomy (AT) and tonsillectomy (TE): does it prevent bleeding complications? AB - BACKGROUND: Bleeding remains the most important complication of adenotonsillectomy in children. Preoperative coagulation tests are widely used to detect unknown bleeding disorders. To determine the efficacy of preoperative coagulation screening in preventing bleeding complications. PATIENTS: Study group 1: 148 healthy children referred by the otorhinolaryngology department for preoperative pediatric examination. Study group 2: 124 healthy children sent to the hemostaseologic clinic for preoperative investigation of a prolonged PTT. METHOD: The incidence of relevant coagulation disorders detected by a standardized bleeding history and coagulation screening tests was studied prospectively in 2 study groups planned for AT and/or TE. The frequency of abnormal bleeding was investigated retrospectively in those children who underwent surgery. RESULTS: Bleeding disorders were detected in 7/148 and 15/124 children in study group 1 and 2 respectively. 141/148 and 79/124 children actually underwent surgery, 62 TE +/- AT + 79 AT alone and 26 TE +/- AT + 53 AT alone respectively. Major bleeding occurred in 1/141 patients (1 TE) in study group 1. Preoperatively, this child had shown normal coagulation screening tests. In 4/79 patients (3 TE, 1 AT) in study group 2, surgery was complicated by major bleeding. Despite extensive testing, no relevant bleeding disorder had been diagnosed in these children preoperatively. Sensitivity of coagulation screening tests for major bleeding was 0 in study group 1. CONCLUSIONS: In our study, coagulation screening failed to effectively identify patients at risk of bleeding. PMID- 17080337 TI - Antiepileptic treatment in paediatric oncology--an interdisciplinary challenge. AB - Epileptic seizures are a common and clinically relevant problem in paediatric oncology. Attributable to the heterogeneity of this group of patients and a number of possible comorbidities antiepileptic treatment in paediatric oncology poses a number of diagnostic and therapeutic challenges. This requires a close interdisciplinary approach to the seizing child or adolescent. A prompt and detailed diagnostic work-up is needed in every case in order to establish the diagnosis and, equally important, to detect secondary aetiological factors, e. g. epileptogenic drugs or any acute underlying pathology, such as metabolic or toxic encephalopathies, CNS-infections or cerebrovascular events. This might offer the opportunity for a specific causative treatment and thus prevent unnecessary long term antiepileptic drug (AED) treatment. If AED treatment is initiated several aspects have to be taken into account. Most importantly, AEDs and chemotherapeutic drugs (CTDs) may interact. Depending on the comedication this may result in reduced tumour or seizure control or unexpected toxicity of AEDs or CTDs. Understanding these interactions will allow to anticipate clinically relevant adverse effects. AED may be further complicated by side-effects, some of them of particular concern for children or adolescents, such as cognitive effects, myelotoxicity, serious rashes, endocrinological disturbances, and many more. Beside critically questioning the need for AED treatment it is therefore important to prefer AED with a good safety-profile in this population. Enzyme inducing and inhibiting AED should be avoided if possible. Preliminary studies indicate that gabapentin and levetiracetam may provide good options in terms of efficacy and safety. However, more properly designed clinical studies are warranted to raise the level of evidence for robust clinical recommendations. Until that time, clinicians will need to continue to question current policies and adapt their daily practice to evolving scientific data. PMID- 17080338 TI - Acute ascending motoric paraplegia following intrathecal chemotherapy for treatment of acute lymphoblastic leukemia in children: case reports and review of the literature. AB - Severe side effects of chemotherapy in pediatric patients with acute lymphoblastic leukemia are rare, but well-known. We present two pediatric patients who developed ascending motoric paraplegia (AMP) following intrathecal chemotherapy. Both patients suffered from progressive weakness of their lower extremities, neurogenic bladder dysfunction, autonomous neural dysregulation and minor sensory deficits. Despite an initially similar clinical picture, progression and outcome were fairly different. There is convincing evidence that AMP is caused by spinal cord toxicity of intrathecally applied toxic agents such as cytarabin and/or methotrexate leading to spinal demyelinisation as demonstrated by elevated myelin basic protein in cerebrospinal fluid. PMID- 17080339 TI - Immunotherapy in children: report from the Reisensburg-Symposium October 20-22, 2004 and recent advances. AB - Immunotherapy of childhood malignancies has a magic aura of promising an easy way to cure. It is not only the public, but also the medical community, which has caused this hope. Therefore, it was worthwhile to bring together a panel of experts working in the field of immunotherapy in order to discuss ongoing projects, and to show hopes and disappointments. The result was a fascinating symposium sponsored by The Kind Philipp Leukemia Foundation (Monchengladbach, Germany). The symposium took place in the Reissensburg near Ulm (Germany) which gives the ideal location for such an event. PMID- 17080340 TI - [Effects of self-adapting G-DRG system 2004 to 2006 on in-patient services payment in pediatric hematology and oncology patients of a university hospital]. AB - BACKGROUND: Reimbursement of inpatient treatment by daily constant charges is replaced by diagnosis- and procedure-related group system (G-DRG) in German acute care hospitals excerpt for psychiatry since 2004. Re-designs of G-DRG system were undertaken in 2005 and 2006. Parallel to implementation requirement- and resource based self-adjustment of this new reimbursement system has been established by law. Adjustments performed in 2005 and 2006 are examined with respect to their effect on reimbursements in treatments of children with oncological, hematological, and immunological diseases. PATIENTS AND METHODS: An unchanged population of 349 patients associated with 1,731 inpatient stays of a Clinic of Pediatric Oncology, Hematology, and Immunology in 2004 was analyzed by methods and means of G-DRG systems 2004, 2005, and 2006. DRGs and additional payments for drugs and procedures eligible for all and/or individual hospitals were calculated. RESULTS: G-DRG system 2005 resulted in overall reimbursement loss of 3.77 % compared to G-DRG 2004. G-DRG 2006 leads to slightly improved overall reimbursements compared to G-DRG 2005 by increasing DRG-based revenues. G-DRG 2006 effects 2.40 % reduction in overall reimbursement compared to G-DRG 2004. This loss includes ameliorating effects of additional payments for drugs and blood products already. Despite introduction of additional payments especially designed for children and teenagers in 2006, additional payment volume is decreased by 21.71 % from 2005 to 2006. G-DRG 2006 yields over-all reimbursement losses of 1.45 % in comparison to G-DRG 2004. Overall reimbursements include introduced additional payments for drugs and blood products. (Reimbursements resulting out of DRG payment alone drop by 14.73 % from 2004 to 2005, and increase by 3.26 % from 2005 to 2006 (2004 vs. 2006 11.95 %). Introduction of additional payments for drugs and blood products on a Germany-wide basis introduced in 2005 dampens DRG-based reimbursement losses. Despite introduction of dosage intervals specifically designed for children and adolescents in 2006, reimbursement of additional payments for drugs and blood products decrease by 21.71 % from 2005 to 2006. An important revenue-balancing function is attributed to additional charges individual for each hospital according to Par. 6 Section 2 (New diagnostic and therapeutic methods) and Section 2 a KHEntgG (German Hospital Reimbursement Law) with respect to financing tertiary care focusses. If possible to attain, those charges may partially equalize losses. Including these additional charges per individual hospital balance of summarized additional charges is -3.89 % from 2005 to 2006. However, fraction of additional payments on total reimbursements increases from 0.64 % in 2004 to 11.98 % in 2005, and 11.24 % in 2006, respectively. CONCLUSIONS: The G-DRG system in its versions 2005 and 2006 results in lowering overall reimbursements of a pediatric hematology, oncology, and immunology department compared to initial status in 2004. The growing chargeability of additional payments ameliorate this effect. PMID- 17080341 TI - [New gelatine-free TBE vaccines with improved safety profile]. PMID- 17080342 TI - [Current treatment of hamate hook fractures]. AB - PURPOSE: Hamate hook fractures can be treated conservatively or surgically, whereby fragment and fracture position, age of the fracture, associated injuries, and individual needs of the patient all influence the choice of the therapeutic procedure. Acute non-displaced fractures are frequently treated conservatively, while displaced fractures and nonunions undergo surgical treatment. We report our experience in diagnostic and therapeutic management of 14 hamate hook fractures. CLINICAL MATERIAL AND METHOD: During a three-year period, 14 patients (eleven men and three women) 21 to 73 years old (42.0 +/- 17.9 years) with fractures at the base of the hamate hook were treated at three hand surgery units. The retrospective study of all patients included a chart review, postoperative radiological imaging after one year, and clinical examinations with grip strength measurements after 18 to 34 months (27.8 +/- 4.9 months). In six patients (43 %), the acute fracture was immobilized in a lower arm cast for six weeks, while eight patients (57 %) were operated primarily. In five cases (36 %) excision of the fragment and in three cases (21 %) open reduction and internal fixation using a screw were performed. Of six patients treated conservatively, five developed nonunion after two to five months (3.0 +/- 1.2 months) with persistent pain and underwent secondary surgery. One patient was asymptomatic despite a nonunion and declined surgical treatment. In three cases the fragment was excised, while two patients underwent open reduction and internal fixation with a screw. RESULTS: All patients operated primarily were free of complaints three months after surgery. The success rate of surgical treatment (8/8) was therefore significantly higher than that after conservative treatment (1/6). CONCLUSION: Compared to conservative treatment of acute non-displaced hamate hook fractures, which is associated with a high risk of developing symptomatic nonunion, primary surgical treatment reliably yields a good clinical outcome. Here, results after fragment excision and open reduction and internal fixation are comparable. PMID- 17080343 TI - [Surgical anatomy of the carpal tunnel]. AB - The carpal tunnel is the pathway between the flexor compartment of the distal forearm and the midpalmar space of the hand. It contains all the long finger and thumb flexor tendons including the median nerve. In cases of a discrepancy between the volume of the carpal tunnel in relation to its contents, high pressures in this region can result. There is a risk that the median nerve will be sustained by compression. The carpal tunnel is covered by the flexor retinaculum. Length and width of the retinaculum correspond to the extension of the tunnel. Radial und ulnar osseous walls can narrow the tunnel diameter as well as an prominent capitate bone at the floor. The median nerve normally courses the tunnel radially near the middle of the flexor retinaculum far reaching straight to the midpalmar region. Several deviations of its tunnel course, atypical divisions, accessory branches and variations of the ramification of the thenar branch of the median nerve are described. PMID- 17080344 TI - [Specific strength measurement of musculus abductor pollicis brevis in order to objectively evaluate muscle regeneration after carpal tunnel release surgery]. AB - 47 patients with a carpal tunnel syndrome were preoperatively examined not only by electrophysical studies (nerve velocity and distal motoric latency) and clinical tests (Phalen's sign, Tinel's sign) but also by measuring the strength of the M. abductor pollicis brevis with a specifically designed strength-testing unit. 30 patients agreed to participate in a voluntary postoperative examination. In the 45 degrees -measurement there was a relative strength loss of the injured hand's thumb of about 25 % in comparison with the healthy hand preoperatively. The postoperative strength disadvantage amounted to only 10 %. Measurement of the strength in the neutral zero method showed a relative strength loss of the injured hand's thumb of 8 % while postoperatively there was even a strength advantage of 9 % in comparison with the thumb of the healthy hand. Both methods showed a relevant and significant postoperative strength increase of the injured hand's thumb. We could prove a significant correlation (p = 0.03) of the neutral zero strength measurement and the distal motoric latency of the median nerve. Furthermore, we could show a trend for a correlation between the neutral zero strength measurement and the median motoric velocity. In conclusion, it can be said that the special strength measurement of the M. abductor pollicis brevis is an important parameter to objectively evaluate the motoric symptomatology in carpal tunnel syndrome patients. PMID- 17080345 TI - [Surgical treatment of persisting and recurrent carpal tunnel syndrome from 1999 to 2003]. AB - The operative treatment of carpal tunnel syndrome is relatively simple and is carried out by doctors from various surgical specialities. In cases of persisting pain or postoperative worsening of the condition, the indication for the procedure could have been wrong or an iatrogenic complication may have to be taken into consideration. We have analysed 42 patients (48 hands) who underwent surgical treatment for carpal tunnel release from 1999 to 2003. We treated eight men and 34 women with an average age of 56 years in this way. Ten patients were initially operated upon by endoscopic release. We found an incompletely transected or even untouched retinaculum flexorum in 16 patients. In eight patients we found an iatrogenic nerve lesion and 24 patients developed serious scarring. Revision surgery should be undertaken only by a surgeon who is a specialist in hand surgery and has extensive experience in this field. In the same context, postoperative hand therapy is essential for a good result, which is performed by occupational therapists in our clinic. PMID- 17080346 TI - [Reoperations for CTS because of recurrence or for correction]. AB - BACKGROUND: With an increasing number of operative procedures for CTS, the number of reoperations is increasing too. These procedures are not in general performed because of recurrence, other reasons may play a role for the failure of the initial operation or recurrent symptoms. METHODS AND CLINICAL MATERIAL: Revision procedures performed in 57 patients in the practice for peripheral neurosurgery from January to September 2004 were analyzed for incision, intraoperative findings, method of operation, electrophysiological findings, and revised diagnosis. Moreover, the data were compared with 185 revision procedures performed in the years from 1986 up to 1994. RESULTS: 2.4 % of all CTS operations were revision procedures (in the former time period 1.5 %). In 59 % of revision operations (former 50 %) an incomplete release of the transverse ligament was found, in 27 % (former 31 %) typical recurrence, in 5 % (former 6 %) nerve lesions, and in 9 % (former 13 %) no abnormalities so that other reasons for complaints of mainly radicular lesions must be assumed. In cases of incomplete release of the transverse ligament, only in 16 % of the patients were normal skin incisions seen, but in 56.3 % there were small incisions (i.e., short or mini incisions). Typical CTS recurrence occurred mainly in hemodialysis patients, nerve lesions were seen mainly when endoscopic procedures were performed. CONCLUSION: Diagnostic problems because of incomplete or misinterpreted ENG findings may lead to delayed or useless primary as well as revision operations. Too small incisions but also endoscopic procedures used by less experienced surgeons are accompanied with an increased risk for avoidable revisions and nerve lesions. Not only for forensic reasons but also in view of quality management, procedures for correction (of operative failure) should be distinguished from those for recurrence. PMID- 17080347 TI - [Re-intervention after carpal tunnel release]. AB - In the last eight years we performed 63 re-interventions because of recurrent carpal tunnel syndromes. Intact parts of the retinaculum were the reason for persistent symptoms in 38 patients. 21 patients suffered from scar tethering, two patients from circumferential fibrosis of the nerve. In one patient a ganglion and in another patient a direct injury to the median nerve was detected. In 21 patients short incisions were performed in the primary procedures. Since the introduction of endoscopic carpal tunnel release, an open decompression via a short incision is recommended as an alternative. Since then we encounter an increase of necessary operative revisions after short incisions. Optical tools may increase the survey especially in short incisions. In extended teno synovialitis of the flexor tendons, a longer incision over the wrist enables adequate conditions for a safe teno-synovialectomy. The length of the incision has to be chosen according to the pathologic-anatomic situation, the disposable equipment and the experience of the surgeon. PMID- 17080348 TI - Protective coverage of the median nerve using fascial, fasciocutaneous or island flaps. AB - PURPOSE: The aim of the study is to present our experience with fascial or fasciocutaneous pedicle and island flaps in the treatment of recurrences of CTS with and without median nerve lesions. MATERIAL AND METHODS: From 1987 to 2006 we have operated on 25 patients (17 women and 8 men, ages ranging from 38 to 76 years with a mean age of 55 years) due to a recurrence of CTS. All the patients required nerve coverage using a local or distant flap. There were 19 hypothenar fat flaps; two forearm radial artery flaps, a forearm ulnar artery flap, an ulnar fascial-fat flap and a posterior interosseous flap. Patients were clinically and instrumentally evaluated before the operation. Assessments of the evaluation parameters were classified in excellent, good, fair and poor according to clinical and return to work criteria. RESULTS: Patients were evaluated after a mean follow-up of 51 months (12 to 168 months). The pain evaluation showed an improvement passing from a mean value of 9 to 4. The best results were for those patients in whom the median nerve was undamaged (mean value of 1). Eleven patients obtained excellent results; good results were obtained in twelve cases; two patients demonstrated fair results due to partial median nerve injury. In these cases, a hypothenar fat flap and an ulnar fascial-fat flap were used, respectively. CONCLUSION: Protective coverage of the median nerve by using fascial or fasciocutaneous flaps after failure of CTR and/or unsuccessful re operations is a good solution to furnish to the median nerve a gliding tissue to avoid adherences with the surrounding tissue of previous surgery. The protection of the nerve can reduce painful symptoms even if it does not permit a return to a painless condition. However, the clinical results in terms of median nerve functional recovery cannot be predicted: if the median nerve is damaged, protective coverage of it by flaps cannot give a favourable result in terms of recovery of both sensory and motor deficits. PMID- 17080349 TI - [The iatrogenic carpal tunnel syndrome--case report]. AB - An inadequate indication for a carpal tunnel revision procedure may be followed by disastrous sequelae for the patient. So it may be justified to talk of iatrogenic CTS. A patient suffering from bilateral CTS is presented who had been operated on six times in all. The following techniques had been used by an orthopedic surgeon, a neurologist, a neurosurgeon and a hand surgeon: microsurgical neurolysis, epineurectomy, neuroma resection, tenosynovectomy and finally a hypothenar fat flap. Initial cause for this fatal series was scar tenderness following lesion of the muscular and palmar branches of the median nerve in the first or second operation. Since sensory nerve conduction was normal after decompression had been performed, there would have been no indication for further surgery of the median nerve. Resection of the neuromas of the two injured branches was not followed by any relief for the patient nor did wrapping the nerve in a fat flap help. Such courses may lead to high costs in health care and occupational disability. They can be avoided by competent neurological and electrophysiological examination, correct interpretation of findings as well as critical consideration of the indication for revision procedures. PMID- 17080350 TI - [The reamer--a new instrument for simplified harvesting of donor bone from the iliac crest]. AB - Harvesting donor bone from the iliac crest site to fill gaps in bone defects is a simple operation but there are still reports of many complications occurring in the bone harvesting process. We now describe a procedure that significantly decreases the morbidity of the donor site. Reports on the iliac crest miller modified according to Krimmer that was applied on 40 patients (average age: 68 years) afforded results that were compared with data collected from the use conventional methods (average age: 65 years). The average length of the skin incision of 41 mm was significantly shorter than the incision length recorded in connection with the conventional methods, namely 79 mm. The harvesting time was limited to 7.6 minutes instead of 18 minutes und thus was also significantly shorter. The postoperative pain as measured on the basis of a visual analogue scale (0 to 100 points) was as follows: pain records were lower than those of the control group by 29 points on the second day following the operation, by 25 points on the fourth day, and by 17 points on the 12th day following the operation. No complications were observed. In the control group there were three haematomas, two cicatricial dehiscences, one cicatricial keloid and one loss of sensibility. This new procedure fulfils the demands for a minimally invasive technique. PMID- 17080351 TI - [Presumptive identification of Candida albicans by rapid diagnostic tests]. AB - PURPOSE OF THE STUDY: The goal of the study was to evaluate the practical value of rapid tests in laboratory diagnostics of pathogenic yeasts with focus on Candida albicans and to elaborate an alternative method of germ tube test in NYP medium. MATERIAL AND METHODS: 200 clinical strains of 12 yeast species isolated in mycology laboratory at the Department of Clinical Microbiology, Teaching Hospital in Hradec Kralove were investigated. All these isolates were studied according to their growth onto the two chromogenic media (CandiSelect, Bio-Rad, HiCrome Candida Agar, HiMedia) and the production of germ tubes in serum and NYP medium. The results of identification by the rapid tests were verified by standard methods. RESULTS: All 100 C. albicans isolates grew on CandiSelect as well as HiCrome Candida agar as characteristically coloured colonies. In case of HiCrome Candida agar, C. tropicalis strains (n = 15) produced typically blue green colonies. Sensitivity and specifity of both the media was 100%. Germ tubes were observed in serum and NYP medium in all C. albicans strains. Lower proportion of pseudohyphae was noticed in NYP medium than serum particularly in non-albicans Candida strains. Two strains C. tropicalis (13.3 %) produced germ tubes in serum, but not NYP. CONCLUSIONS: Both CandiSelect and HiCrome Candida Agar identified reliably C. albicans isolates, in case of the latter C. tropicalis as well. The comparison of the production of germ tubes in serum versus NYP medium showed comparable results and lower risk of false positive results in NYP medium which can follow from pseudohyphae formation. All rapid tests studied represent reliable tools for presumptive identification of C. albicans. PMID- 17080352 TI - [Can the determination of IgG antibodies to the pertussis toxin help the diagnosis of the disease?]. AB - BACKGROUND: In the differential diagnosis of protracted irritating cough we should always consider the possibility of pertussis. Serology performed primarily in a late stage of the disorder does not always provide a clear answer. We wanted to verify whether a quantitative determination of IgG antibodies to the pertussis toxin (IgG-PT) could help establish a clear diagnosis. MATERIAL AND METHODS: Between 1 January and 30 June 2005 we performed serological investigations in 139 children presenting with an irritable cough or after application of an acellular pertussis vaccine. In 95 children we compared the serological response of agglutination antibodies and IgG-PT. RESULTS: After vaccination the children presented with different levels of antibodies and these were not always identical in two types of serological response. Children with clinical manifestations of pertussis presented a good formation of antibodies, but the two kinds of antibodies often persisted for long periods at unchanged levels. CONCLUSIONS: The determination of IgG-PT can assist the diagnosis of the disease, but this investigation alone cannot yield a clear-cut confirmation of pertussis. PMID- 17080353 TI - [Staining of mycobacteria without pre-heating in a modification of the Ziehl Neelsen method]. AB - BACKGROUND: To evaluate the use of a new solution for the staining of mycobacteria without pre-heating for microscopic examinations in visible light. To verify the possibilities of the method in everyday practice. MATERIAL AND METHODS: For the diagnostic staining of mycobacteria we used a modification of the Ziehl-Neelsen method without pre-heating. The carbolfuchsin solution contained a higher proportion of basic fuchsin, alcohol and phenol than in the original method; all other solutions used in this method remained unchanged. Efficacy was verified on 8 mycobacterial strains by a comparison with the original procedure that uses pre-heating, as well as with the fluorescence method. 310 microscopic specimens from clinical samples were compared with the fluorescence method, 2 500 smears from cultures were compared with the original staining method. RESULTS: The results of mycobacteria staining with a modification of the Ziehl-Neelsen method without pre-heating were similar as those obtained by staining with the original method. The sensitivity of the new method was slightly lower than fluorescence (89 % and 92 %), but its specificity was higher (82 % and 65 %). CONCLUSIONS: The method can be used for the staining of mycobacteria in everyday practice. Its efficacy was verified on microscopic specimens from clinical samples and on smears from cultures. PMID- 17080354 TI - [The demonstration of tuberculosis using molecular genetic methods polymerase chain reaction (PCR) and Amplified Mycobacterium tuberculosis direct (AMTD) test]. AB - INTRODUCTION: Tuberculosis is a communicable disease, in most instances with a chronic course. The aetiological agent is Mycobacterium tuberculosis. Its demonstration is based on microscopic investigations and cultures. Microscopy is not sufficiently sensitive, while cultures are lengthy. One of the possibilities of speeding up diagnosis are molecular genetic methods. PURPOSE OF THE STUDY: To compare the demonstration of mycobacteria using molecular genetic methods with the results of cultures. METHODS: We used two methods to demonstrate the nucleic acid complex of Mycobacterium tuberculosis-the polymerase chain reaction (PCR) and the Amplified Mycobacterium tuberculosis direct test (AMTD). We investigated 647 samples. Out of these, 275 samples were tested with PCR and 372 were investigated with a AMTD set. At the same time we started for each sample a parallel culture. RESULTS: In 275 samples, out of a total of 647, which were analysed with PCR, mycobacterial DNA was demonstrated in 18 (6.5 %). Out of the 372 samples investigated with AMTD, mycobacterial RNA was demonstrated in 27 (7 %). Out of the 18 PRC positive samples, 6 (13 %) did not yield a positive mycobacterial culture. Out of the 27 positive results RNA with the AMTD method 17 did not yield positive cultures. On the other hand, a diagnosis of tuberculosis verified by cultures without a positive PCR was found in 2 patients (0.7 %). Disagreement between the results of AMTD and cultures was also found in 2 samples (0.5 %). CONCLUSIONS: Molecular genetic methods substantially speed up the diagnosis of tuberculosis. These methods are particularly important in cases of paucibacillary material and of unique and unrepeatable samples (tissues biopsies, nodes, cerebrospinal fluid). Given the possibility of false positive results, parallel verification by microscopy and cultures is essential. PMID- 17080355 TI - [Pulmonary sarcoidosis--a rare complication of interferon treatment for chronic hepatitis C]. PMID- 17080356 TI - Scale-dependent inhibition drives regular tussock spacing in a freshwater marsh. AB - Regular spatial patterning is common in nature, and various mechanisms of self organization have been proposed to explain regular patterning. We report on regular spatial patterning in Carex stricta in a freshwater wetland and investigate the applicability of theoretical models that explain regular patterning based on inhibition, facilitation, or interaction between the two. Spectral analysis of aerial photographs revealed that tussocks were regularly spaced at an average distance of 60 cm. Photosynthetically active radiation varied significantly with distance from the tussock and was lowest at intermediate distance from the tussock center (15-40 cm). Using transplants to assay growth conditions, we found that C. stricta grew well in all distance classes with and without natural C. stricta biomass, except at intermediate distances when buried in C. stricta wrack. Our experimental results reveal that C. stricta inhibits its growth in a scale-dependent manner: inhibition was found to peak at intermediate distance from the tussock. We compared three alternative models to examine potential mechanisms driving regularity and found that, similar to our experimental results, scale-dependent inhibition provides the best explanation for the observed regular tussock spacing. Our study underlines the importance of scale-dependent feedback in the formation of regular spatial patterning in ecosystems. PMID- 17080357 TI - Evolutionary predictions should be based on individual-level traits. AB - Recent theoretical studies have analyzed the evolution of habitat specialization using either the logistic or the Ricker equation. These studies have implemented evolutionary change directly in population-level parameters such as habitat specific intrinsic growth rates r or carrying capacities K. This approach is a shortcut to a more detailed analysis where evolutionary change is studied in underlying morphological, physiological, or behavioral traits at the level of the individual that contribute to r or K. Here we describe two pitfalls that can occur when such a shortcut is employed. First, population-level parameters that appear as independent variables in a population dynamical model might not be independent when derived from processes at the individual level. Second, patterns of covariation between individual-level traits are usually not conserved when mapped to the level of demographic parameters. Nonlinear mappings constrain the curvature of trade-offs that can sensibly be assumed at the population level. To illustrate these results, we derive a two-habitat version of the logistic and Ricker equations from individual-level processes and compare the evolutionary dynamics of habitat-specific carrying capacities with those of underlying individual-level traits contributing to the carrying capacities. Finally, we sketch how our viewpoint affects the results of earlier studies. PMID- 17080358 TI - Evolutionary and ecological causes of the latitudinal diversity gradient in hylid frogs: treefrog trees unearth the roots of high tropical diversity. AB - Why are there more species in the tropics than in temperate regions? In recent years, this long-standing question has been addressed primarily by seeking environmental correlates of diversity. But to understand the ultimate causes of diversity patterns, we must also examine the evolutionary and biogeographic processes that directly change species numbers (i.e., speciation, extinction, and dispersal). With this perspective, we dissect the latitudinal diversity gradient in hylid frogs. We reconstruct a phylogeny for 124 hylid species, estimate divergence times and diversification rates for major clades, reconstruct biogeographic changes, and use ecological niche modeling to identify climatic variables that potentially limit dispersal. We find that hylids originated in tropical South America and spread to temperate regions only recently (leaving limited time for speciation). There is a strong relationship between the species richness of each region and when that region was colonized but not between the latitudinal positions of clades and their rates of diversification. Temperature seasonality seemingly limits dispersal of many tropical clades into temperate regions and shows significant phylogenetic conservatism. Overall, our study illustrates how two general principles (niche conservatism and the time-for speciation effect) may help explain the latitudinal diversity gradient as well as many other diversity patterns across taxa and regions. PMID- 17080359 TI - Functional significance of shade-induced leaf senescence in dense canopies: an experimental test using transgenic tobacco. AB - Canopy photosynthesis models have predicted an optimal leaf area index (LAI; leaf area per unit surface area) and leaf nitrogen distribution at which whole-plant carbon gain per unit N is maximized. In this study we experimentally tested these models, using transgenic P(SAG12)-IPT tobacco (SAG; Nicotiana tabacum L.) plants with delayed leaf senescence and therefore a greater LAI and more uniform N distribution than the wild type (WT). In a competition experiment, the increased density of surrounding WT plants caused a greater reduction in dry mass of mature SAG target plants than in that of WT target plants, indicating negative effects of delayed leaf senescence on performance at high canopy density. Vegetative SAG plants achieved a lower calculated daily carbon gain than competing WT plants because the former retained leaves with a negative carbon gain in the shaded, lower part of the canopy. Sensitivity analyses showed that the carbon gain of SAG plants would increase if these lower leaves were shed and the N reallocated from these leaves were used to form additional leaf area at the canopy top. This strategy, which is adopted by the WT, is most advantageous because it results in the shading of competing neighbors. PMID- 17080360 TI - Plant reproductive allocation predicts herbivore dynamics across spatial and temporal scales. AB - Life-history theory suggests that iteroparous plants should be flexible in their allocation of resources toward growth and reproduction. Such plasticity could have consequences for herbivores that prefer or specialize on vegetative versus reproductive structures. To test this prediction, we studied the response of the cactus bug (Narnia pallidicornis) to meristem allocation by tree cholla cactus (Opuntia imbricata). We evaluated the explanatory power of demographic models that incorporated variation in cactus relative reproductive effort (RRE; the proportion of meristems allocated toward reproduction). Field data provided strong support for a single model that defined herbivore fecundity as a time varying, increasing function of host RRE. High-RRE plants were predicted to support larger insect populations, and this effect was strongest late in the season. Independent field data provided strong support for these qualitative predictions and suggested that plant allocation effects extend across temporal and spatial scales. Specifically, late-season insect abundance was positively associated with interannual changes in cactus RRE over 3 years. Spatial variation in insect abundance was correlated with variation in RRE among five cactus populations across New Mexico. We conclude that plant allocation can be a critical component of resource quality for insect herbivores and, thus, an important mechanism underlying variation in herbivore abundance across time and space. PMID- 17080361 TI - Host-parasite coevolution and selection on sex through the effects of segregation. AB - The advantage of producing novel variation to keep apace of coevolving species has been invoked as a major explanation for the evolution and maintenance of sex (the Red Queen hypothesis). Recent theoretical investigations of the Red Queen hypothesis have focused on the effects of recombination in haploid species, finding that species interactions rarely favor the evolution of sex unless selection is strong. Yet by focusing on haploids, these studies have ignored a potential advantage of sex in diploids: generating novel combinations of alleles at a particular locus through segregation. Here we investigate models of host parasite coevolution in diploid species to determine whether the advantages of segregation might rescue the Red Queen hypothesis as a more general explanation for the evolution of sex. We find that the effects of segregation can favor the evolution of sex but only under some models of infection and some parameter combinations, almost always requiring inbreeding. In all other cases, the effects of segregation on selected loci favor reductions in the frequency of sex. In cases where segregation and recombination act in opposite directions, we found that the effects of segregation dominate as an evolutionary force acting on sex in diploids. PMID- 17080362 TI - Testing the beneficial acclimation hypothesis and its alternatives for locomotor performance. AB - The beneficial acclimation hypothesis (BAH) is controversial. While physiological work all but assumes that the BAH is true, recent studies have shown that support for the BAH is typically wanting. The latter have been criticized for assessing the benefits of developmental plasticity rather than acclimation. Here we examine the BAH within a strong inference framework for five congeneric species of ameronothroid oribatid mites that occupy marine to terrestrial habitats. We do so by assessing responses of maximum speed, optimum temperature, and performance breadth, measured from -10 degrees C to 35 degrees C, to four treatment temperatures (0 degrees , 5 degrees , 10 degrees , and 15 degrees C). We show that the BAH and its alternatives often make similar empirical predictions. Weak beneficial acclimation is characteristic of one of the more marine species. In the other two upper-shore and marine species, evidence exists for deleterious acclimation and the colder-is-better hypothesis. In the two fully terrestrial species, there is no plasticity. Lack of plasticity is beneficial when cue reliability is low or costs of plasticity are high, and the former seems plausible in terrestrial habitats. However, weak plasticity in the upper shore/marine species and the absence of plasticity in the terrestrial species might also be a consequence of phylogenetic constraint. PMID- 17080363 TI - The effects of switching behavior on the evolutionary diversification of generalist consumers. AB - Mathematical models of consumer-resource systems explore the evolution of a morphological trait that determines two resource acquisition rates in a generalist consumer. The consumer also has the ability to adjust its relative consumption of the two resources via behavioral (or developmental) plasticity subject to a trade-off. The analysis examines both stable systems and those with sustained fluctuations in abundance. In both cases, it seeks to determine how the behavioral choice affects the evolution of the morphological characters. The presence of adaptive switching behavior transforms the shape of the relationship between the morphological character and fitness in a manner that usually leads to evolution of two or more morphological types. As in models without switching, the presence of sustained cycles in resource densities often allows the evolution of a generalist as well as two specialists. However, switching expands and shifts the parameter regions yielding this outcome and in some cases allows the evolution and coexistence of at least two generalists as well as the two specialists. This level of diversity supported by only two resources is not seen in the absence of behavioral choice and resource cycles. The results suggest major roles for both behavior and environmental variation in adaptive radiation. PMID- 17080364 TI - Pleistocene rewilding: an optimistic agenda for twenty-first century conservation. AB - Large vertebrates are strong interactors in food webs, yet they were lost from most ecosystems after the dispersal of modern humans from Africa and Eurasia. We call for restoration of missing ecological functions and evolutionary potential of lost North American megafauna using extant conspecifics and related taxa. We refer to this restoration as Pleistocene rewilding; it is conceived as carefully managed ecosystem manipulations whereby costs and benefits are objectively addressed on a case-by-case and locality-by-locality basis. Pleistocene rewilding would deliberately promote large, long-lived species over pest and weed assemblages, facilitate the persistence and ecological effectiveness of megafauna on a global scale, and broaden the underlying premise of conservation from managing extinction to encompass restoring ecological and evolutionary processes. Pleistocene rewilding can begin immediately with species such as Bolson tortoises and feral horses and continue through the coming decades with elephants and Holarctic lions. Our exemplar taxa would contribute biological, economic, and cultural benefits to North America. Owners of large tracts of private land in the central and western United States could be the first to implement this restoration. Risks of Pleistocene rewilding include the possibility of altered disease ecology and associated human health implications, as well as unexpected ecological and sociopolitical consequences of reintroductions. Establishment of programs to monitor suites of species interactions and their consequences for biodiversity and ecosystem health will be a significant challenge. Secure fencing would be a major economic cost, and social challenges will include acceptance of predation as an overriding natural process and the incorporation of pre-Columbian ecological frameworks into conservation strategies. PMID- 17080365 TI - Multilocus test for introgression between the cactophilic species Drosophila mojavensis and Drosophila arizonae. AB - Information obtained from laboratory studies regarding the efficacy of barriers to gene flow (reproductive isolation) between species is often incomplete or misleading, so detailed genetic analyses are needed to determine whether hybridization and introgression occur in nature. Previous laboratory studies of the cactophilic species Drosophila mojavensis and Drosophila arizonae suggest that reproductive isolation is incomplete and that gene flow may occur in sympatry. We sampled 18 nuclear and one mitochondrial loci from multiple populations of D. arizonae and D. mojavensis to test for the signature of recent or historic gene flow between these two species. We located chromosomal regions that were inverted between these species and analyzed those regions independently of others. Statistical tests for introgression using all loci or only collinear loci failed to reject expectations of an isolation model. Further tests using average nucleotide differences between species and phylogenetic analyses also failed to find support for introgression between D. mojavensis and D. arizonae. Additional ecological and behavioral studies of these species in their natural habitats are required to explain why the signature of gene flow was not detected at the DNA sequence level in populations when laboratory studies suggest such gene flow should be possible. PMID- 17080366 TI - Floater dynamics can explain positive patterns of density-dependent fecundity in animal populations. AB - After some 70 years of debate on density-dependent regulation of animal populations, there is still poor understanding of where spatial and temporal density dependence occurs. Clearly defining the portion of the population that shapes density-dependent patterns may help to solve some of the ambiguities that encircle density dependence and its patterns. In fact, individuals of the same species and population can show different dynamics and behaviors depending on their locations (e.g., breeding vs. dispersal areas). Considering this form of intrapopulation heterogeneity may improve our understanding of density dependence and population dynamics in general. We present the results of individual-based simulations on a metapopulation of the Spanish imperial eagle Aquila adalberti. Our results suggest that high rates of floater mortality within settlement areas can determine a shift in the classical relationship (from negative to positive) between the fecundity (i.e., fledglings per pair) and density (i.e., number of pairs) of the breeding population. Finally, we proved that different initial conditions affecting the breeder portion of the population can lead to the same values of fecundity. Our results can represent a starting point for new and more complex approaches studying the regulation of animal populations, where the forgotten and invisible component--the floater--is taken into account. PMID- 17080367 TI - On testing the competition-colonization trade-off in a multispecies assemblage. AB - The competition-colonization trade-off has long been considered an important mechanism explaining species coexistence in spatially structured environments, yet data supporting it remain ambiguous. Most competition-colonization research examines plants and the dispersal-linked traits of their seeds. However, colonization is more than just dispersal because rapid population growth is also an important component of colonization. We tested for the presence of competition colonization trade-offs with a commonly used artificial assemblage consisting of protozoan and rotifer species, where colonization was the ability of a species to establish populations in patches. By ranking species according to their colonization abilities and their pairwise competitive interactions, we show that these species exhibit competition-colonization trade-offs. These results reveal that the competition-colonization trade-off exists within nonplant assemblages and that even in a laboratory setting, species are constrained to be either good competitors or colonizers but not both. PMID- 17080368 TI - Outbreak investigations: red-eyed rabbits and community service. PMID- 17080369 TI - Nationwide outbreak of red eye syndrome associated with transfusion of leukocyte reduced red blood cell units. AB - OBJECTIVE: To characterize red eye reactions occurring within 24 hours after receipt of units of leukocyte-reduced red blood cells, determine their etiology, and investigate their potential link to transfusion. METHODS: We conducted a survey of transfusion facilities nationwide to determine the scope and magnitude of the reactions; performed case-control and cohort studies among transfused patients at the facility where most reactions occurred; and performed animal experiments, using cellulose acetate derivatives extracted from leukocyte reduction filters and filter precursors, to reproduce reactions. RESULTS: From January 1, 1997, through January 15, 1998, we identified 159 reactions in 117 patients from 17 states. Reactions were characterized by conjunctival erythema or hemorrhage (in 100% of patients), eye pain (in 62%), photophobia (in 46%), and decreased visual acuity (in 32%). Symptom onset occurred 1-24 hours after initiation of transfusion and resolved within a median of 5 days. Reactions were associated with transfusion sessions that included units of red blood cells filtered with a specific brand of filter, the LeukoNet filter (HemaSure) (odds ratio, 100.4; P<.001). There was a dose-response relationship between the number of LeukoNet-filtered units transfused and the attack rate for reactions, ranging from 0.8% among sessions in which 1 unit was transfused to 27.3% among sessions in which 3 or more units were transfused (P<.001). A similar ocular syndrome was elicited in rabbits injected with cellulose acetate derivatives extracted from unused LeukoNet filters or filter precursors. No reactions were reported after LeukoNet filters were withdrawn from the market. CONCLUSIONS: This transfusion associated red eye syndrome was linked to a specific brand of leukocyte-reduction filter and likely resulted from cellulose acetate derivatives leached from the filter membrane. PMID- 17080370 TI - Outbreak of Serratia marcescens colonization and infection traced to a healthcare worker with long-term carriage on the hands. AB - OBJECTIVE: To reveal the source of a nosocomial outbreak of colonization and infection with a strain of Serratia marcescens positive for Guiana extended spectrum beta-lactamase 1 (GES-1) that occurred among patients in a neurosurgical intensive care unit (ICU) in a Dutch university medical center from May 2002 through March 2003. METHODS: Samples from the environment and from the hands of healthcare workers (HCWs) were cultured. A retrospective case-control study was carried out. RESULTS: Fifteen neurosurgical ICU patients who had 1 or more cultures that yielded the epidemic strain of S. marcescens from May 2002 through March 2003 were defined as case patients and matched with 30 control patients. Environmental cultures did not reveal a prominent source of S. marcescens. Cultures of specimens from the hands of 100 HCWs revealed colonization of a single HCW with the epidemic strain. Although this HCW instantly went on leave, serial cultures detected prolonged carriage of the epidemic strain on the hands of the HCW for 3 months. The skin of the HCW's hands was psoriatic. The epidemic abruptly ended after the colonized HCW went on leave. Retrospective case-control analysis showed that the patients colonized or infected with S. marcescens received significantly more nursing care from the colonized HCW than did control patients (P<.05). From February 2004 through October 2004, a second cluster of 3 patients was detected with the epidemic strain of S. marcescens. In October 2004, the formerly colonized HCW appeared to have carriage of the epidemic strain on the hands again. CONCLUSIONS: A single HCW with the epidemic strain of S. marcescens on the hands was considered the source of this outbreak. PMID- 17080371 TI - Transmission of group A Streptococcus limited to healthcare workers with exposure in the operating room. AB - BACKGROUND: Nosocomial transmission of group A Streptococcus (GAS) has been well described. A recent report of an outbreak investigation suggested that transmission can be extensive and that standard infection control measures may not be adequate to prevent transmission from patients with severe, invasive disease to healthcare workers (HCWs). OBJECTIVE: A case of pharyngitis in an HCW caring for a patient with GAS pharyngitis and necrotizing fasciitis prompted an investigation of the extent and risk factors for nosocomial transmission of GAS. SETTING: A 509-bed, tertiary care center in Portland, Oregon with 631,100 patient visits (hospital and clinic) and 11,500 employees in the year 2003. METHODS: HCWs with exposure to the index patient ("contacts") were identified for streptococcal screening and culture and completion of a questionnaire regarding the location and duration of exposure, use of personal protective equipment, and symptoms of GAS infection. RESULTS: We identified 103 contacts of the index patient; 89 (86%) submitted oropharyngeal swabs for screening and culture. Only 3 (3.4%) of contacts had a culture that yielded GAS; emm typing results and pulsed-field gel electrophoresis patterns of GAS isolates from 2 HCWs were identical to those for the isolate from the index patient. Both HCWs were symptomatic, with febrile pharyngitis and reported prolonged contact with the open wound of the patient in the operating room. CONCLUSIONS: In this investigation, nosocomial transmission was not extensive, and standard precautions provided adequate protection for the majority of HCWs. Transmission was restricted to individuals with prolonged intraoperative exposure to open wounds. As a result, infection control policy for individuals was modified only for HCWs with exposure to GAS in the operating room. PMID- 17080372 TI - Outbreak of bloodstream infection with the mold Phialemonium among patients receiving dialysis at a hemodialysis unit. AB - BACKGROUND: Molds are a rare cause of disseminated infection among dialysis patients. OBJECTIVE: We evaluated a cluster of intravascular infections with the mold Phialemonium among patients receiving hemodialysis at the same facility in order to identify possible environmental sources and prevent further infection. DESIGN: Environmental assessment and case-control study. SETTING: A hemodialysis center affiliated with a tertiary care hospital. METHODS: We reviewed surveillance and clinical microbiology records and performed a blood culture survey for all patients. The following data for case patients were compared with those for control patients: underlying illness, dialysis characteristics, medications, and other possible exposure for 120 days prior to infection. Environmental assessment of water treatment, dialysis facilities, and heating, ventilation, and air-conditioning (HVAC) systems of the current and previous locations of the dialysis center was performed. Samples were cultured for fungus; Phialemonium isolates were confirmed by sequencing of DNA. Investigators observed dialysis access site disinfection technique. RESULTS: Four patients were confirmed as case patients, defined as a patient having intravascular infection with Phialemonium species; 3 presented with fungemia, and 1 presented with an intravascular graft infection. All case patients used a fistula or graft for dialysis access, as did 12 (75%) of 16 of control patients (P=.54). Case and control patients did not differ in other dialysis characteristics, medications received, physiologic findings, or demographic factors. Phialemonium species were not recovered from samples of water or dialysis machines, but were recovered from the condensation drip pans under the blowers of the HVAC system that supplied air to the dialysis center. Observational study of 21 patients detected suboptimal contact time with antiseptic agents used to prepare dialysis access sites. CONCLUSION: The report of this outbreak adds to previous published reports of Phialemonium infection occurring in immunocompromised patients who likely acquired infection in the healthcare setting. Recovery of this mold from blood culture should be considered indicative of infection until proven otherwise. Furthermore, an investigation into possible healthcare-related environmental reservoirs should be considered. PMID- 17080373 TI - Outbreak of Chlamydia pneumoniae infection in a Japanese nursing home, 1999-2000. AB - OBJECTIVE: To identify risk factors for infection and severe illness due to Chlamydia pneumoniae. METHODS: To identify risk factors for infection, we conducted a case-control study among nursing home residents who had onset of symptoms during December 1, 1999, to February 20, 2000. To identify risk factors for severe illness among nursing home residents, we conducted a retrospective cohort study. SETTING: A nursing home providing long-term and day care services for elderly patients in Japan.Participants. Fifty-nine residents and 41 staff members of a nursing home. RESULTS: The attack rates for respiratory illness were 53% (31 of 59) among residents and 22% (9 of 41) among staff. Infection was confirmed in 15 resident and 2 staff case patients by isolation of C. pneumoniae from nasal swab specimens. Fifteen resident case patients developed severe illness (ie, bronchitis, pneumonia, and hypoxia); one case patient died. The median age of resident case patients was 87 years. We could identify neither the source of the outbreak nor significant risk factors for infection and severe illness in residents. However, residents with a higher level of physical activity were more likely to become infected, whereas older residents (aged more than 85 years) and those with a lower level of physical activity were more likely to develop severe illness (P>.05). Contact with residents was a risk factor for infection in staff (relative risk, undefined; P=.04). CONCLUSIONS: C. pneumoniae can cause large outbreaks of infection and severe illness among elderly persons, and its transmission is likely to be enhanced by close contacts among people in nursing homes. Therefore, early detection of an outbreak by means of better surveillance, and subsequent isolation of patients, may be effective control measures. PMID- 17080374 TI - Seroprevalence survey of varicella, measles, rubella, and hepatitis A and B viruses in a multinational healthcare workforce in Saudi Arabia. AB - OBJECTIVE: To determine the prevalence of the vaccine-preventable diseases caused by varicella, measles, rubella, and hepatitis A and B viruses in a multinational healthcare workforce. DESIGN: Prospective cohort study. SETTING: A 750-bed tertiary care center located in Riyadh, Saudi Arabia. METHODS: In compliance with hospital policy, newly recruited healthcare workers (HCWs) were enrolled in the study from September 2001 to March 2005. Serum samples were collected from all HCWs during the initial hiring process and tested for IgG antibodies against each of the 5 viral agents. Nonimmune HCWs were subsequently vaccinated at the earliest opportunity. RESULTS: A total of 4,006 newly hired (international and local) employees were included in the study. All underwent serologic testing for IgG antibodies against varicella, measles, rubella, hepatitis A, and hepatitis B viruses. Of the total, 63% were female and 37% were male. Middle Eastern employees comprised 47% of the total, followed by employees from the Far East (35%), the West (10%), and Africa (8%). Forty-two percent were nurses, 27% were in administration, 18% were medical technicians, and 13% were physicians. Among the 4,006 newly hired HCWs, 14% had negative IgG antibody test results for varicella virus, 13% for measles virus, 10% for rubella virus, 33% for hepatitis A virus, and 43% for hepatitis B virus. More women than men were susceptible to hepatitis A (40% vs. 24%; P<.001), whereas more men were susceptible to hepatitis B (55% vs. 35%; P<.001). Varicella susceptibility was more common among HCWs from the Far East (19%), whereas susceptibility to measles, rubella, hepatitis A, and hepatitis B was highest among HCWs from the Middle East. Both relative youth and male sex were associated with lack of antibodies against hepatitis B virus and rubella virus. In contrast, female sex and younger age were associated with lack of antibodies against hepatitis A virus (P<.001). CONCLUSION: Seroprevalence surveys of vaccine-preventable diseases among HCWs, although labor intensive, are invaluable in caring for a multinational workforce. PMID- 17080375 TI - Optimal bandaging of smallpox vaccination sites to decrease the potential for secondary vaccinia transmission without impairing lesion healing. AB - OBJECTIVE: To assess the optimal method for covering smallpox vaccination sites to prevent transmission of vaccinia. DESIGN: Randomized, nonblinded clinical trial. SETTING: Tertiary care medical center. PARTICIPANTS: Vaccinia-naive and vaccinia-experienced volunteers. INTERVENTIONS: After vaccination, study participants were randomized to receive 1 of 3 types of bandage: gauze, occlusive with gauze lining, or foam. Vaccination sites were assessed every 3 to 5 days until the lesion healed. During each visit, specimens were obtained from the vaccination site, the bandage surface before removal, and the index finger contralateral to the vaccination site and were cultured for vaccinia. Time to lesion healing was assessed. RESULTS: All 48 vaccinia-naive and 47 (87%) of 54 vaccinia-experienced participants developed a vesicle or pustule at the injection site 6-11 days after vaccination. Fourteen (14%) of 102 participants had bandage cultures positive for vaccinia. All but 1 of these vaccinia-positive cultures were of a bandage from participants randomized to the gauze bandage group, and all but 3 were of bandages from vaccinia-naive participants. No finger-specimen cultures were positive for vaccinia. One episode of neck autoinoculation occurred in a vaccinia-naive individual who had vaccinia recovered from his gauze bandage on multiple visits. The foam bandage was associated with more local adverse effects (skin irritation and induration). The time to healing did not differ among the bandage groups. CONCLUSIONS: The potential for transmission of vaccinia from a vaccination site is greater if the site is covered by gauze than if it is covered by occlusive or foam bandages. Use of an occlusive bandage with a gauze lining is the best choice for coverage of smallpox vaccination sites because of a reduced potential for vaccinia transmission and a lower reactogenicity rate. Bandage choice did not affect vaccination lesion healing. PMID- 17080376 TI - Experimental evaluation of an automated endoscope reprocessor with in situ generation of peracetic acid for disinfection of semicritical devices. AB - OBJECTIVE: To evaluate the effectiveness of a high-level disinfection solution generated inside an endoscope processing system for decontaminating external and internal surfaces of experimentally contaminated heat-sensitive medical devices. METHODS: The American Society for Testing and Materials Simulated-Use Test protocol (E1837-02), which incorporates a soil load in each inoculum, was used to evaluate the efficacy of the system when processing 4 common types of endoscopes contaminated separately with 5 types of nosocomial pathogens: Pseudomonas aeruginosa (ATCC 15442), spores of Clostridium difficile (ATCC 9689), a glutaraldehyde-resistant strain of Mycobacterium chelonae, a vancomycin-resistant strain of Enterococcus faecalis, and a methicillin-resistant strain of Staphylococcus aureus. Rinse solution samples from channels and from surfaces of the processed endoscopes were tested for any microbicidal residues. RESULTS: For all organisms tested, the baseline level of contamination of the endoscopes ranged from 5 log(10) to greater than 7 log(10) at each external surface site and internal channel. All tests showed reductions in viability of the test organisms to undetectable levels. All rinse solution samples from external and internal sites of the endoscopes proved to be free of any residual microbicidal activity. CONCLUSIONS: The endoscope reprocessor, with its processor-generated high-level disinfection solution, successfully reduced the numbers of selected, clinically relevant pathogens to undetectable levels both in the channels and on the outside surfaces of the 4 representative endoscopes tested in this study. PMID- 17080377 TI - Management of an outbreak of Clostridium difficile-associated disease among geriatric patients. AB - OBJECTIVE: To describe a nosocomial outbreak of Clostridium difficile-associated disease (CDAD). DESIGN: A traditional outbreak investigation. SETTING: Geriatric department of a tertiary care teaching hospital from March through April 2003. METHODS: The outbreak was detected by the C. difficile surveillance program of the infection control unit. CDAD was diagnosed by stool culture and fecal toxin A detection with a qualitative rapid immunoassay. Isolates of C. difficile were serotyped and genotyped using pulsed-field gel electrophoresis. RESULTS: The incidence of CDAD increased from 27 cases per 100,000 patient-days in the 6-month period before the outbreak to 99 cases per 100,000 patient-days during the outbreak. This outbreak involved 21 of 92 patients in 4 geriatric wards, which were located at 2 geographically distinct sites and staffed by the same medical team. The mean age of patients was 83 years (range, 71-100 years). Five (24%) of the 21 patients had community-acquired diarrhea, and secondary hospital transmission resulted in 3 clusters involving 16 patients. Serotyping and genotyping were performed on isolates in stool specimens from 19 different patients; 16 of these isolates were serotype A1, whereas 3 displayed profiles different from the outbreak strain. Management of this outbreak consisted in reinforcement of contact isolation precautions for patients with diarrhea, cohorting of infected patients in the same ward, and promotion of hand hygiene. Relapses occurred in 6 (29%) of 21 patients. CONCLUSION: Control of this rapidly developing outbreak of CDAD was obtained with early implementation of cohorting and ward closure and reinforcement of environmental disinfection, hand hygiene, and enteric isolation precautions. PMID- 17080378 TI - Duration of methicillin-resistant Staphylococcus aureus carriage, according to risk factors for acquisition. AB - OBJECTIVE: To examine the duration of methicillin-resistant Staphylococcus aureus (MRSA) carriage and its determinants and the influence of eradication regimens. DESIGN: Retrospective cohort study. SETTING: A 1,033-bed tertiary care university hospital in Bern, Switzerland, in which the prevalence of methicillin resistance among S. aureus isolates is less than 5%. PATIENTS: A total of 116 patients with first-time MRSA detection identified at University Hospital Bern between January 1, 2000, and December 31, 2003, were followed up for a mean duration of 16.2 months. RESULTS: Sixty-eight patients (58.6%) cleared colonization, with a median time to clearance of 7.4 months. Independent determinants for shorter carriage duration were the absence of any modifiable risk factor (receipt of antibiotics, use of an indwelling device, or presence of a skin lesion) (hazard ratio [HR], 0.20 [95% confidence interval {CI}, 0.09-0.42]), absence of immunosuppressive therapy (HR, 0.49 [95% CI, 0.23-1.02]), and hemodialysis (HR, 0.08 [95% CI, 0.01 0.66]) at the time MRSA was first MRSA detected and the administration of decolonization regimen in the absence of a modifiable risk factor (HR, 2.22 [95% CI, 1.36-3.64]). Failure of decolonization treatment was associated with the presence of risk factors at the time of treatment (P=.01). Intermittent screenings that were negative for MRSA were frequent (26% of patients), occurred early after first detection of MRSA (median, 31.5 days), and were associated with a lower probability of clearing colonization (HR, 0.34 [95% CI, 0.17-0.67]) and an increased risk of MRSA infection during follow-up. CONCLUSIONS: Risk factors for MRSA acquisition should be carefully assessed in all MRSA carriers and should be included in infection control policies, such as the timing of decolonization treatment, the definition of MRSA clearance, and the decision of when to suspend isolation measures. PMID- 17080379 TI - community-acquired infection with healthcare-associated methicillin-resistant Staphylococcus aureus: the role of home nursing care. AB - OBJECTIVE: To better understand the role of indirect transmission in community acquired infection with methicillin-resistant Staphylococcus aureus (MRSA). DESIGN: Prospective case-control study. SETTING: A French teaching hospital. PATIENTS: A total of 198 case patients and 198 control patients with MRSA or methicillin-susceptible S. aureus infection diagnosed between April 2002 and July 2003. RESULTS: Multivariate analysis showed a highly significant independent link between MRSA infection at admission and prior receipt of home nursing care (odds ratio [OR], 3.7; P<.001). Other independent risk factors were prior hospitalization (OR, 3.8; P<.001), transfer from another institution (OR, 2.3; P=.008), and age older than 65 years (OR, 1.6; P=.04). Prior home nursing care showed a frequency dose-response relationship. Eleven MRSA-infected patients had had home nursing procedures but no hospital stay in the previous 3 years. These patients' MRSA strains were related to the prevalent MRSA clone currently spreading in French hospitals. CONCLUSION: Home nursing care appears to be an independent risk factor for MRSA acquisition in the community. The reservoir probably consists of MRSA carriers discharged from the hospital. Community nurses seem to be a potential vector. PMID- 17080380 TI - Staphylococcus aureus bacteremia in older adults: predictors of 7-day mortality and infection with a methicillin-resistant strain. AB - OBJECTIVES: To determine the predictors of 7-day mortality in older adult patients with Staphylococcus aureus bacteremia after controlling for comorbidity using the Charlson weighted index of comorbidity (WIC) and to identify the risk factors associated with bacteremia due to methicillin-resistant S. aureus (MRSA). DESIGN: Retrospective cohort study from January 2003 until December 2004. SETTING: Two tertiary care, university-affiliated hospitals. METHODS: One hundred thirty-five hospitalized patients with S. aureus bacteremia were included in the study. All patients who were 60 years or older and had 1 or more blood cultures positive for S. aureus were included in the study. The primary outcome was death 7 days after the onset of S. aureus bacteremia. RESULTS: Twenty-one patients (15.6%) died within 7 days after the onset of S. aureus bacteremia. Seventy-four patients (56.1%) had MRSA bacteremia. Multivariate analysis identified 3 independent determinants of 7-day mortality: Charlson WIC score greater than 5 (odds ratio [OR], 3.6 [95% confidence interval {CI}, 1.1-11.2]; P=.03), previous hospitalization in the past 3 months (OR, 5.0 [95% CI, 1.1-25.1]; P=.04), and altered mental status at the onset of S. aureus bacteremia (OR, 13.6 [95% CI, 2.9 64.6]; P=.001). Multivariate analysis identified previous hospitalization in the past 3 months (OR, 2.6 [95% CI, 1.1-5.9]; P=.02), residence in a long-term care facility (OR, 4.5 [95% CI, 1.7-12.3]; P=.003), and altered mental status at the onset of S. aureus bacteremia (OR, 2.5 [95% CI, 1.5-5.6]; P=.02) to be independently associated with the presence of MRSA. CONCLUSIONS: The Charlson WIC is significantly associated with increased mortality of S. aureus bacteremia in older adults. Previous hospitalization in the past 3 months, residence in a long term care facility, and altered mental status should be used as a guidance for empirical vancomycin therapy and application of infection control measures in older adults with suspected S. aureus bacteremia. PMID- 17080381 TI - Impact of extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella species on clinical outcomes and hospital costs: a matched cohort study. AB - OBJECTIVES: To evaluate the economic and clinical impact of infection with extended-spectrum beta -lactamase (ESBL)-producing Escherichia coli and Klebsiella species (ESBL-EK). DESIGN: A matched-cohort analysis of the cost of illness. SETTING: An 810-bed, urban, community hospital in Hartford, Connecticut. PATIENTS: Twenty-one case patients infected with ESBL-EK at a site other than the urinary tract were matched with 21 control subjects infected with a non-ESBL producing organism on the basis of pathogen species, age, anatomic site of infection, hospitalization in the intensive care unit (ICU) during the time of infection, date of hospitalization, and initial antibiotics received. RESULTS: Mean infection-related costs per patient were significantly greater for case patients than for control patients ($41,353 vs $24,902; P=.034). Infection related length of stay was the main driver of cost, which was prolonged for case patients, compared with control patients (21 vs 11 days; mean difference, 9.7 days [95% confidence interval {CI}, 3.2-14.6 days]; P=.006). The additional cost attributed to the presence of an ESBL-EK infection was $16,450 per patient (95% CI, $965-$31,937). Case patients were more likely than control patients to have clinical failure (P=.027), and the rate of treatment success for case patients whose initial treatment involved antibiotics other than carbapenems was lower than that for their matched control patients (39% vs 83%; P=.013). Treatment was successful in patients for whom initial treatment was with a carbapenem, regardless of the ESBL status of the pathogen. CONCLUSION: The cost of non urinary tract infections caused by ESBL-EK was 1.7 times the cost of non-urinary tract infections caused by non-ESBL producers. Prompt recognition and appropriate antimicrobial selection may minimize this ESBL-related impact on hospital costs. PMID- 17080382 TI - Screening for multidrug-resistant bacteria as a predictive test for subsequent onset of nosocomial infection. AB - OBJECTIVE: To investigate whether carriage of multidrug-resistant bacteria is a risk factor for nosocomial infection and whether detection of carriage is predictive of subsequent onset of nosocomial infection. METHODS: In this observational cohort (study period, June 1998 through October 2002), nasal and rectal swab specimens from 412 consecutive patients admitted to the intensive care unit were tested for carriage of multidrug-resistant bacteria. Concomitantly, the bacteria responsible for any subsequent nosocomial infection, the date of infection, and some of the known clinical risk factors for nosocomial infection were noted. These factors were adjusted for potential confounders, using a Cox model stratified on the propensity score of multidrug-resistant bacteria carriage. The diagnostic characteristics of a carriage test, including the positive and negative diagnostic likelihood ratios, were calculated for all strata of the propensity score. RESULTS: Forty-two patients were carrying multidrug-resistant bacteria. Nosocomial infection occurred in 95 patients, of whom 16 (38%) were carriers, and 79 (83%) were noncarriers (P=.01). After adjustment for potential confounders, statistical analysis revealed that carriage remained a risk factor for nosocomial infection (relative risk, 2.08 [95% confidence interval {CI}, 1.13-3.81]). Receipt of antibiotic treatment at the time of intensive care unit admission was found to be protective against nosocomial infection. A positive result of test for detection of carriage seemed to be an efficient predictor of subsequent nosocomial infection (positive diagnostic likelihood ratio, 2.05 [95% CI, 1.15-3.66]), although a negative test result was not a predictor of subsequent nosocomial infection (negative likelihood ratio, 0.91 [95% CI, 0.73-1.11]). CONCLUSION: Carriage proved to be a risk factor for subsequent nosocomial infection. However, the carriage test was useful as a predictive tool only for patients with a positive test result. PMID- 17080383 TI - Programmatic factors related to smallpox vaccine uptake by healthcare workers and others. AB - We surveyed program coordinators at 106 hospitals and health departments that participated in the National Smallpox Vaccination Program to ascertain how program-level factors affected the rate of smallpox vaccine uptake by staff. In a fully adjusted multivariate model, health departments achieved significantly higher vaccination rates than did hospitals, as did facilities that invited fewer employees to be vaccinated. PMID- 17080384 TI - Outbreak of endophthalmitis after cataract surgery: the importance of the quality of the surgical wound. AB - Postoperative endophthalmitis is a rare but potentially devastating condition. We investigated an outbreak of 8 cases of endophthalmitis in patients who underwent phakectomy performed by a single surgeon from January through September 2004. The outbreak was traced to damaged surgical blades, and it highlights the importance of the quality of the surgical wound. PMID- 17080385 TI - Nosocomial outbreak of Pseudomonas aeruginosa endophthalmitis. AB - We describe an outbreak of nosocomial endophthalmitis due to a common source, which was determined to be trypan blue solution prepared in the hospital's pharmacy service. We assume that viable bacteria probably gained access to the trypan blue stock solution during cooling after autoclaving. The temporal cluster of Pseudomonas aeruginosa endophthalmitis was readily perceived on the basis of clinical and microbiological findings, and an exogenous source of contamination was unequivocally identified by means of DNA fingerprinting. PMID- 17080386 TI - An outbreak of Pneumococcal pneumonia among residents of a retirement home in France during October 2003. AB - We describe an outbreak of pneumonia due to Streptococcus pneumoniae in a French retirement home. Eleven residents developed pneumonia. Eight patients had positive results of urinary antigen tests. There were no further cases after the implementation of control measures, which involved isolation of and receipt of antibiotic therapy by symptomatic residents. No risk factors for transmission of S. pneumoniae were identified in this population. PMID- 17080387 TI - A cluster of nosocomial herpes simplex virus type 1 pneumonia in a medical intensive care unit. AB - We report a cluster of 3 cases of nosocomial herpes simplex virus type 1 (HSV-1) pneumonia occurring in close temporal and physical proximity during a 1-week period, which suggested a common source. HSV-1 nosocomial pneumonia occurs in immunocompetent intubated patients and presents as otherwise unexplained profound and/or prolonged hypoxemia (decreased F(IO2), increased P(O2), and decreased A-a gradient) and "failure to wean." The diagnosis of HSV-1 pneumonia is determined by demonstration of characteristic cytopathologic findings (Cowdry type A inclusion bodies) in distal respiratory epithelial cells from bronchoscopic specimens. Acyclovir therapy results in rapid improvement and ability to wean. PMID- 17080388 TI - Pseudo-outbreak of hepatitis B virus infection associated with contamination of a semiautomatic cap remover. AB - A pseudo-outbreak of hepatitis B virus caused by cross-contamination from a semiautomatic cap remover for blood collection tubes is reported. The source of the outbreak was elucidated by using basic epidemiological methods. Laboratories should always be critical about their results in order to identify contamination problems. PMID- 17080389 TI - Ethnicity and other possible risk factors for candidemia at 3 tertiary care university hospitals in Hawaii. AB - A total of 108 cases of candidemia detected in 3 tertiary care university hospitals in Hawaii between January 2001 and December 2002 were retrospectively reviewed. Candida albicans, Candida glabrata, and Candida tropicalis accounted for 28% of the cases. Mortality among Filipino patients was significantly higher than that among other ethnic groups (71% vs 48%; P<.05). PMID- 17080390 TI - Comparative cost of selective screening to prevent transmission of methicillin resistant Staphylococcus aureus (MRSA), compared with the attributable costs of MRSA infection. AB - The annual cost of a screening program to detect methicillin-resistant Staphylococcus aureus (MRSA) in a teaching hospital in Spain was 10,261 Euro. The average cost per MRSA infection was 2,730 Euro; therefore, the cost of the program would be covered if it only prevented 4 infections per year (11% of the total number of MRSA infections at our hospital). PMID- 17080391 TI - Methicillin-resistant Staphylococcus aureus survival on hospital fomites. AB - We examined the duration of survival of 2 strains of methicillin-resistant Staphylococcus aureus (MRSA) on 3 types of hospital fomites. MRSA survived for 11 days on a plastic patient chart, more than 12 days on a laminated tabletop, and 9 days on a cloth curtain. Irregular surfaces may help harbor organisms in the environment. In addition to contact precautions, MRSA containment during an outbreak should include concurrent environmental decontamination. PMID- 17080392 TI - Clinical culture surveillance of carbapenem-resistant Pseudomonas aeruginosa and Acinetobacter species in a teaching hospital in Sao Paulo, Brazil: a 7-year study. AB - Carbapenem-resistant Pseudomonas aeruginosa and Acinetobacter species are worrisome nosocomial pathogens. After introduction of a preventive program involving clinical surveillance culture to reduce the spread of those pathogens, we observed an 80% decrease in the percentage of cultures that yielded carbapenem resistant Acinetobacter isolates. The percentage of cultures that yielded carbapenem-resistant P. aeruginosa remained relatively stable during the intervention. PMID- 17080393 TI - Knowledge, attitudes, and behaviors regarding piperacillin-tazobactam prescribing practices: results from a multicenter study. AB - We investigated knowledge, attitudes, and behaviors of prescribers concerning piperacillin-tazobactam use at 4 Emory University-affiliated hospitals. Discussions during focus groups indicated that the participants' perceived knowledge of clinical criteria for appropriate piperacillin-tazobactam use was inadequate. Retrospective review of medical records identified inappropriate practices. These findings have influenced ongoing interventions aimed at optimizing piperacillin-tazobactam use. PMID- 17080394 TI - Factors that influence influenza vaccination among healthcare workers in a French geriatric hospital. AB - In a geriatric hospital in France, only 80 (21%) of 390 healthcare workers (HCWs) were vaccinated against influenza. Predictive factors for accepting influenza vaccination were occupation as a physician (odds ratio [OR], 9.79), previous receipt of influenza vaccination (OR, 36), and desire to protect their own health (OR, 2.42) and residents' health (OR, 3.68). Predictive factors for refusing influenza vaccination were occupation as a nurse (OR, 6.41) or nursing assistant (OR, 4.04) and belief that homeopathic medication is more effective than the vaccine (OR, 5.75). PMID- 17080395 TI - Intrinsically contaminated alcohol-free mouthwash implicated in a nosocomial outbreak of Burkholderia cepacia colonization and infection. PMID- 17080396 TI - Analysis of costs attributable to an outbreak of severe acute respiratory syndrome at a French hospital. PMID- 17080397 TI - Defining influenza-like illness. PMID- 17080398 TI - Markers of absorption and synthesis of cholesterol in men with type 1 diabetes. AB - BACKGROUND AND METHODS: Serum cholestanol and plant sterol ratios to cholesterol, surrogate markers of cholesterol absorption, are assumed to be high in type 1 diabetes (T1D), and the ratios of cholesterol precursor sterols (markers of synthesis) are assumed to be low reflecting downregulated cholesterol synthesis. To this end, we measured serum sterols with gas-liquid-chromatography in 56 men with T1D and in 18 controls to evaluate cholesterol metabolism. Subjects were categorised into tertiles by the cholestanol to cholesterol ratio of controls indicating low to high absorption of cholesterol. RESULTS: The ratios of the synthesis markers were negatively related to the absorption markers in controls, but less consistently in T1D. The absorption markers were positively related to each other, but interrelation of the synthesis markers was less consistent in T1D. In the low absorbers the absorption markers were higher in T1D than in controls (e.g. sitosterol ratio 173 +/- 9 in T1D vs 135 +/- 11 10(2) x mmol/mol of cholesterol in controls, p < 0.05). In the high absorbers, the absorption markers were similar in T1D and controls, but the synthesis markers were higher in T1D than in controls (e.g. lathosterol ratio 154 +/- 10 in T1D vs 120 +/- 5 10(2) x mmol/mol of cholesterol in controls, p < 0.05). CONCLUSIONS: Absorption and synthesis of cholesterol are less closely related to each other in T1D than in controls, but the markers of cholesterol absorption are interrelated also in T1D. Absorption of cholesterol is higher in T1D than in controls within the range of low absorption, but similar in those with relatively high cholesterol absorption. PMID- 17080399 TI - High levels of carcinogenic polycyclic aromatic hydrocarbons (PAH) in 20 brands of Chinese cigarettes. AB - Tar and nicotine levels have been made to conform to EU standards as of 1 July 2004, but data on tobacco-derived carcinogenic compounds, such as PAH, in Chinese cigarettes are lacking in the literature. Levels of tar, nicotine, carbon monoxide and PAH were measured in 20 cigarette brands purchased in China between 2003 and 2004. Higher nicotine and tar levels were found in Chinese cigarettes than in European brands just 3 months before the above deadline; carcinogenic PAH levels were about 1.5 fold higher than in European cigarettes, but analysed singly, the mean value of benzo(a)pyrene (B(a)P) and dibenzo(a,h)anthracene (DBA), the most potent carcinogenic PAH yields, were 2.4 and 4.4 fold higher, respectively. Tar levels were well correlated with carcinogenic PAH (r = 0.53, P < 0.01), thus providing an easily measurable parameter for ranking various cigarette brands in developing countries where more sophisticated techniques might not be feasible for lack of funds and expertise. PMID- 17080400 TI - S-adenosylmethionine or 5'-methylthioadenosine are unable to prevent fumonisin B1 hepatotoxicity in mice despite increased oxidation in liver. AB - Fumonisins are mycotoxins produced by the fugus Fusarium verticillioides, a common fungus growing on corn. Fumonisin B(1) (FB(1)) is the most toxic and prevalent fumonisin detected in corn and corn-based foods. It produces species-, gender-specific damage, and is hepatotoxic and nephrotoxic in rodents. Disruption of sphingolipid metabolism resulting from inhibition of ceramide synthase leads to alterations of cell signaling events, particularly tumor necrosis factor (TNF)alpha signal pathways and to the toxic effects of FB(1). It has been reported that FB(1) toxicity involves oxidative stress. S-adenosylmethionine (SAM) and methylthioadenosine (MTA), an intermediate metabolite in SAM metabolism, are hepatoprotective by modulating TNFalpha expression and increasing reduced glutathione (GSH) levels. The current study investigated the effects of SAM and MTA on FB(1) hepatotoxicity in C57BL/6N mice. The animals were given SAM or MTA by intraperitoneal injection of 25 mg kg(-1) body weight every 12 h when they received subcutaneous injection of 2.25 mg FB(1) kg(-1) body weight once daily for 5 days. The results showed that neither SAM nor MTA protected FB(1) induced liver damage indicated by the increases in activities of plasma alanine aminotransferase and aspartate aminotransferase as well as the number of apoptotic hepatocytes. Both agents prevented an increase of free sphingosine but not sphinganine. Neither SAM nor MTA modified the FB(1)-induced expression of TNFalpha, interleukin (IL)-1alpha or IL-1 receptor antagonist. The decreased GSH in liver following FB(1) treatment was not protected by either agent. The data indicate that SAM and MTA are ineffective in protecting against FB(1) toxic effects. PMID- 17080401 TI - 4-Aminobiphenyl N-glucuronidation by liver microsomes: optimization of the reaction conditions and characterization of the UDP-glucuronosyltransferase isoforms. AB - 4-Aminobiphenyl (4-ABP) is an arylamine that has long been associated with human and animal urinary bladder cancer. N-glucuronidation is an important metabolic pathway that contributes significantly to 4-ABP-bladder carcinogenesis by facilitating transport of the active metabolites from the liver to the bladder. This pathway is carried out by UDP-glucuronosyltransferase (UGTs). These enzymes are located in the inner membrane of the endoplasmic reticulum. Full UGT activity is not achieved until membrane constraints are removed. This study was conducted to optimize the incubation conditions of 4-ABP N-glucuronidation. The kinetic parameters of the isozymes most commonly involved in arylamine glucuronidation, namely UGT1A4 and UGT1A9, were also determined. The UGT reaction was linear in the incubation time (0-90 min) and in the microsomal protein range of 0-0.5 mg. Alamethicin, a pore-forming agent, was found to be the best reagent to activate UGTs. It increased the enzyme activity by nearly 8-fold and this activation was at concentration of 50 microg mg(-1) protein. Interestingly, UGT1A4 glucuronidated 4-ABP with more affinity and efficiency than did UGT1A9. The K(m) and V(max) of UGT1A4 for 4-ABP were 58.8 microm and 234.9 pmol min(-1) mg(-1) protein, respectively, and 227.5 microm and 31.2 pmol min(-1) mg(-1) protein for UGT1A9. Furthermore, hecogenin was found to be a competitive inhibitor for UGT1A4. It increased the K(m) of UGT1A4 for 4-ABP by nearly 10 fold at a concentration of 50 microm. This is the first report that tried to optimize the incubation conditions for 4-ABP N-glucuronidation and characterized the enzyme kinetic parameters of UGT isoforms catalysing 4-ABP N-glucuronidation. PMID- 17080402 TI - Comparison of organic and inorganic mercury distribution in suckling rat. AB - Thiomersal is used as a preservative in vaccines given to small children. The metabolic product of thiomersal is ethylmercury and its distribution and kinetics are still not known, especially at this early age. The purpose of this study was to compare the body distribution of two forms of mercury: organic (thiomersal) and inorganic (mercury(2+) chloride) in very young, suckling rats. Mercury was applied subcutaneously three times during the suckling period on days 7, 9 and 11 of pups age, imitating the vaccination of infants. A single dose of mercury was equimolar in both exposed groups, i.e. 0.81 micromol Hg kg(-1). At 14 days of age the animals were killed and the total mercury analysed in blood and organs (kidney, liver and brain). The analytical method applied was total decomposition, amalgamation, atomic absorption spectrometry. The results showed that the level of mercury was higher in the liver and kidney of the inorganic mercury group than in the thiomersal exposed group. However, the brain and blood concentrations of mercury were higher in the thiomersal exposed group. These results need to be clarified by additional data on the kinetic pathways of ethylmercury compared with inorganic mercury. PMID- 17080403 TI - Mycotoxin identification in moldy dwellings. AB - The objective of this study was to assess the level of macrocyclic trichothecenes (MCT), one of the most potent mycotoxins, on wall surfaces, floor dust and air samples from moldy dwellings. The study was based on an index group comprising 15 flooded dwellings contaminated by Stachybotrys chartarum or Chaetomium and a control group comprising nine dwellings without molds on visual inspection and mold sampling. Three samples were collected from each dwelling: a sample from the moldy wall, using a swab, a floor dust sample on a 0.5 m(2) surface with a cloth and an air sample using a pump comprising a 1 microm Teflon filter. The MCT level was measured using a monoclonal, antibody-based ELISA test. Compared with measurements performed in index dwellings, higher MCT values were measured on floor dust samples from moldy dwellings (P = 0.02). Samples from wall surfaces demonstrated nearly significant differences (P = 0.06). No significant differences were observed for air samples (P = 0.15), but some samples showed increased MCT levels. Significant correlations were observed between the levels measured on wall surfaces, floor dust and air samples (P = 0.02 to 0.05). In contrast, no correlation was observed between moldy surface and MCT levels. In conclusion, this paper reports for the first time direct evidence for the presence of MCT in moldy dwellings. PMID- 17080404 TI - Steroidogenic gene expression in H295R cells and the human adrenal gland: adrenotoxic effects of lindane in vitro. AB - The focus on the refinement, reduction and replacement of animal use in toxicity testing requires the development of cell-based systems that mimic the effects of xenobiotics in human tissues. The human adrenocortical carcinoma cell line, H295R, has been proposed as a model for studies on adrenal steroidogenesis and its disruption. In this study, expression profiles for nine adrenal steroidogenic genes were characterized in H295R cells using real-time RT-PCR. Treatment with forskolin increased cortisol secretion and stimulated transcription of all the steroidogenic genes except SULT2A1. The transcript profile from H295R cells in the presence and absence of forskolin was compared with the transcript profile from human adrenal glands. The gene expression pattern observed in the forskolin treated H295R cells was more similar to that in the human adrenal gland, than the expression pattern in untreated cells. To examine H295R cells as a possible in vitro system for the assessment of adrenal disruption using molecular endpoints, the insecticide lindane (gamma-hexachlorocyclohexane) was used. In vivo, lindane has been shown to inhibit testicular, ovarian and adrenal steroidogenesis. It was demonstrated that lindane reduced cortisol secretion, downregulated the expression of a subset of the genes encoding steroidogenic enzymes and repressed transcriptional activation of the steroidogenic acute regulatory protein (StAR) gene promoter. Thus the H295R cell line provides a good in vitro system for the analysis of the human adrenal steroidogenic pathway at the level of hormone production and gene expression. This in vitro test can be used for the rapid detection of adrenal endocrine disruption and as a tool for mechanistic studies. PMID- 17080405 TI - Protective effects of isothiocyanates towards N-nitrosamine-induced DNA damage in the single-cell gel electrophoresis (SCGE)/HepG2 assay. AB - The aim of this study was to investigate the protective effect of isothiocyanates towards N-nitrosamine-induced DNA damage in the single-cell gel electrophoresis (SCGE)/HepG2 assay. None of the isothiocyanates (ITCs) concentrations tested in the presence or absence of formamidopyrimidine-DNA glycosylase (Fpg), caused DNA damage per se. Combined treatments of HepG2 cells with phenethyl isothiocyanate (PEITC), allyl isothiocyanate (AITC) or indol-3-carbinol (I3C) and N nitrosopyrrolidine (NPYR) or N-nitrosodimethylamine (NDMA) reduced the genotoxic effects of the N-nitrosamines in a dose-dependent manner. The protective effect of the three ITCs tested was higher towards NPYR-induced oxidative DNA damage than against NDMA. The greatest protective effect towards NPYR-induced oxidative DNA damage was shown by I3C (1 microm, 79%) and by PEITC (1 microm, 67%) and I3C (1 microm, 61%) towards NDMA (in the presence of Fpg enzyme). However, in the absence of Fpg enzyme, AITC (1 microm, 72%) exerted the most drastic reduction towards NPYR-induced oxidative DNA damage, and PEITC (1 microm, 55%) towards NDMA. The results indicate that ITCs protect human-derived cells against the DNA damaging effect of NPYR and NDMA, two carcinogenic compounds which occur in the environment. PMID- 17080406 TI - Acrylonitrile-induced extracellular signal-regulated kinase (ERK) activation via protein kinase C (PKC) in SK-N-SH neuroblastoma cells. AB - Acrylonitrile (ACN) is classified by IARC as a probable carcinogen. Chronic exposure to ACN increases the incidence of tumors in various organs of test animals, including the brain and lung. ERK1/2 activation plays crucial roles in cell proliferation and is involved in many steps of tumor progression. Therefore, this study examined whether ACN altered the activation state of ERK1/2 in human neuroblastoma SK-N-SH cells. Treatment of these cells with ACN greatly increased phosphorylation of ERK1/2 in dose- and time-dependent manners. This effect was inhibited by PD 98059 and U 0126, specific inhibitors of MEK, indicating that MEK, an upstream activator of ERK1/2, was directly involved in ACN-induced ERK1/2 activation. Furthermore, the activation of ERK1/2 by ACN was attenuated by inhibition of PKC with GF 109203X, rottlerin and prolonged incubation with PMA (phorbol 12-myristate 13-acetate). This demonstrated the participation of PKC in the ACN-stimulated activation of ERK1/2. Taken together, our results indicate that ACN-induced ERK1/2 activation involves PKC through a MEK-dependent pathway. PMID- 17080407 TI - Toxicological effects of Carica papaya seed extract on spermatozoa of mice. AB - Investigations were carried out on epididymal spermatozoa of male mice to study the effects of high dosages (100, 200 and 300 mg kg(-1) bwt) of aqueous Carica papaya seed extract. The results revealed a significant dosedependent suppression of cauda epididymal sperm motility coinciding with a decrease in sperm count and viability. When tested 45 days after the withdrawal of treatment, complete normalcy was restored, proving that the induced effects were transient. PMID- 17080408 TI - Effects of stabilizers on the destabilization of proteins upon adsorption to aluminum salt adjuvants. AB - Excipients for protein-based vaccines are currently identified by evaluating the stability of the protein in solution. In most cases, however, the protein is adsorbed to the surface of an aluminum salt adjuvant in the final vaccine formulation. Previous studies showed that model protein antigens may be structurally altered and less thermally stable upon adsorption to aluminum salt adjuvants [Jones LS, Peek LJ, Power J, Markham A, Yazzie B, Middaugh CR, 2005, J Biol Chem 280:13406-13414]. The work presented herein provides evidence that compounds that stabilize the protein in solution also stabilize the adsorbed protein; however, the stability of the adsorbed protein in the presence of the stabilizer remains lower than that of the protein in solution. Potential implications of the reduced stability on the approach used to select excipients during formulation development are discussed. PMID- 17080409 TI - Drug release properties of polymer coated ion-exchange resin complexes: experimental and theoretical evaluation. AB - Although ion-exchange resins have been used widely as drug delivery systems, their exact release kinetics has not been reported yet. Usually only the rate limiting step has been taken into account and the rest of the steps have been ignored as instantaneous processes. To investigate the exact release kinetics of polymer-coated drug/ion-exchange resin complexes for sustained drug delivery, the results of new mathematical modeling were compared with experimental results. Drug/resin complexes with a model drug, dextromethorphan, were prepared and used as cores for fluid-bed coating. An aqueous colloidal dispersion of poly(vinyl acetate) was applied for the coating. A comprehensive mathematical model was developed using a mechanistic approach by considering diffusion, swelling, and ion-exchange processes solved by numerical techniques. The rate-limiting factor of the uncoated resin particles was diffusion through the core matrix. Similarly, in the coated particles the rate-limiting factor was diffusion through the coating membrane. The mathematical model has captured the phenomena observed during experimental evaluations and the release dynamics from uncoated and coated (at different coat levels) particles were predicted accurately (maximum RMSE 2.4%). The mathematical model is a useful tool to theoretically evaluate the drug release properties from coated ion-exchange complexes thus can be used for design purposes. PMID- 17080410 TI - A relational database for management of flow cytometry and ELISpot clinical trial data. AB - BACKGROUND: Although relational databases are widely used in bioinformatics with deposited and finalized data, they have not received widespread usage among immunologists for managing raw laboratory data such as that generated by ELISpot or flow cytometry assays. Almost no published guidance exists for immunologists to design appropriate and useful data management systems. METHODS: We describe the design and implementation of a Microsoft Access relational database used in a clinical trial in which the primary immunogenicity measures were ELISpot and intracellular cytokine staining. RESULTS: Our data management system enabled us to perform sophisticated queries and to interpret our data as quantitatively as possible. It could easily be used without modification by other researchers using automated plate reading of ELISpot plates or four color flow cytometry. CONCLUSION: We illustrate in detail the use of a flexible data management system for two of the most widely used immunological techniques. Minor modifications for more colors or other outputs can easily be implemented. Based on this example, other modifications could be easily envisaged for any other quantitative output. PMID- 17080411 TI - Improving the loading and release of NSAIDs from pHEMA hydrogels by copolymerization with functionalized monomers. AB - Poly(hydroxyethyl methacrylate), pHEMA, hydrogels are widely used for preparing implants, contact lenses, and other biomedical devices, which in many circumstances should load drugs to deliver them in the adjacent tissues. To enhance the potential of pHEMA hydrogels as nonsteroidal anti-inflammatory drugs (NSAIDs) delivery systems, 4-vinyl-pyridine (VP) and N-(3-aminopropyl) methacrylamide (APMA) were incorporated to the network (25-150 mM). The incorporated monomers did not change the viscoelastic properties neither the state of water, but remarkably increased the amount of ibuprofen (up to 10-fold) and diclofenac (up to 20-fold) loaded. Dried loaded pHEMA-APMA and pHEMA-VP hydrogels quickly swelled in water but ionic/hydrophobic interactions prevented the amount of drug released to be above 10%. By contrast, once the water-swollen hydrogels were transferred to pH 5.8 or 8.0 phosphate buffers or NaCl solutions, the release was prompted by competition with ions of the medium. The remaining of hydrophobic interactions and the high polymeric density of the pHEMA hydrogels contributed to sustain the release process for at least 24 h for ibuprofen and almost 1 week for diclofenac. The release rate was independent of the salt content and pH in the physiological range of values, which enables the design of hydrogel-based delivery systems with predictable release rate. PMID- 17080412 TI - Influence of polymorphic form, morphology, and excipient interactions on the dissolution of carbamazepine compacts. AB - To gain a deeper understanding of the behavior of carbamazepine (CBZ) and CBZ dihydrate (DH) compacts during in vitro dissolution tests various factors were investigated: hydrate formation of CBZ, crystal morphology, surface area, and excipient influence. Dissolution tests were performed in three different dissolution media: distilled water, hydroxypropyl methylcellulose (HPMC), and polyethylene glycol (PEG) solutions. For the CBZ compacts, the dissolution rate of CBZ in water was fastest (0.338 mg L(-1) min(-1)). With increasing ability of the excipients to inhibit the hydration of CBZ (PEG < HPMC), surprisingly the dissolution rate of CBZ compacts decreased: PEG solution (0.314 mg L(-1) min(-1)) > HPMC solution (0.257 mg L(-1) min(-1)). This implies that DH formation resulted in an apparent increase in the dissolution rate rather than slowing it down. For the DH compacts, the dissolution rate in water (0.055 mg L(-1) min(-1)) was slower than that of PEG and HPMC solutions (0.174 and 0.178 mg L(-1) min(-1), respectively). The contact angle measurements showed a significantly higher value in water (61.0 degrees) than in PEG and HPMC solutions (44.8 degrees and 43.1 degrees, respectively). Although the dissolution of CBZ and DH compacts in various dissolution media are complex processes, the influence and relative importance of these factors were clearly detected providing better understanding of the dissolution behavior of the drug. PMID- 17080413 TI - Thermally associating polypeptides designed for drug delivery produced by genetically engineered cells. AB - Thermally associating polymers, including gelatin, cellulose ethers (e.g., Methocels and poloxamers (e.g., Pluronics) have a long history of use in pharmacy. Over the past 20 years, significant advances in genetic engineering and the understanding of protein secondary and tertiary structures have been made. This has led to the development of a variety of polypeptides that do not occur naturally but can be expressed in recombinant cells and have useful properties that lend themselves to novel applications where current materials cannot perform. The most intensively studied motifs are derived from the consensus repeats of elastin and silk, as well as coiled-coil helices. Many of these designed polypeptides or 'artificial proteins' are thermally associating materials. This property can be exploited to develop solid dosage forms, injectable drug delivery systems, micro- or nanoparticle drug carriers, triggered or targeted release systems, or as a means of simplifying the purification process and thus reducing costs of production of these materials. This review focuses on the development and characterization of this novel class of biomaterials and examines their potential for pharmaceutical applications. PMID- 17080414 TI - Assessing the antifungal activity, pharmacokinetics, and tissue distribution of amphotericin B following the administration of Abelcet and AmBisome in combination with caspofungin to rats infected with Aspergillus fumigatus. AB - The purpose of this study was to assess the antifungal activity, pharmacokinetics, and tissue distribution of amphotericin B (AmpB) following the administration of Abelcet and AmBisome alone and in combination with Caspofungin to rats infected with Aspergillus fumigatus. Aspergillus fumigatus inoculum (2.1 2.5 x 10(7) colony forming units [CFU]) was injected via the jugular vein; 48 h later male albino Sprague-Dawley rats (350-400 g) were administered either a single intravenous (i.v.) dose of Abelcet (5 mg AmpB/kg; n = 6), AmBisome (5 mg AmpB/kg; n = 6), Caspofungin (3 mg/kg; n = 5), Abelcet (5 mg AmpB/kg) plus Caspofungin (3 mg/kg) (n = 6), AmBisome (5 mg AmpB/kg) plus Caspofungin (3 mg/kg) (n = 7), or physiologic saline (non-treated controls; n = 6) once daily for 4 days. Antifungal activity was assessed by organ CFU concentrations and plasma galactomannan levels. Plasma and tissue samples were taken from each animal for AmpB pharmacokinetic analysis and tissue distribution determinations. Abelcet treatment significantly decreased total fungal CFU concentrations recovered in all the organs added together by 73% compared to non-treated controls. Ambisome treatment significantly decreased total fungal CFU concentrations recovered in all the organs added together by 69% compared to non-treated controls. Caspofungin treatment significantly decreased total fungal CFU concentrations recovered in all the organs added together by 80% compared to non-treated controls. Abelcet plus Caspofungin treatment significantly decreased total fungal CFU concentrations recovered in all the organs added together by 81% compared to non-treated controls. Ambisome plus Caspofungin treatment significantly decreased total fungal CFU concentrations recovered in all the organs added together by 98% compared to non-treated controls. Abelcet treatment significantly decreased plasma galactomannan levels by 50 and 75% 96 h following the initiation of treatment in the absence and presence of Caspofungin co-therapy, respectively. AmBisome treatment significantly decreased plasma galactomannan levels by 73 and 78% 96 h following the initiation of treatment in the absence and presence of Caspofungin co-therapy, respectively. Co-administration of Caspofungin with Abelcet and AmBisome did not significantly alter the plasma concentration-time profile, pharmacokinetic parameters, and tissue distribution of AmpB. Taken together, our findings suggest that an alternative mechanism, possibly at the cellular level rather than altered AmpB disposition, may be an explanation for the differences in organ CFU concentrations following Abelcet plus Caspofungin versus AmBisome plus Caspofungin administration. PMID- 17080415 TI - A statistical comparison of pad numbers versus pad weights in the quantification of urinary incontinence. AB - AIMS: Pad per day (PPD) usage is a frequently utilized measure of urinary incontinence. The 24-hour pad weight test (24PWT) is a reproducible test for quantifying incontinence volumes. We investigated whether PPD validly reports the magnitude of urinary incontinence. METHODS: This was a retrospective review of patients undergoing stress incontinence surgery from July 2002 to 2005. Inclusion criteria were a documented 24PWT and patient-reported PPD usage. Grams of urine loss per pad (GPP) provided a third measure of incontinence. Descriptive statistics and correlations between all variables and significance were noted. Factor analysis was performed on the three measures of leakage and age for all patients over age 50. RESULTS: One hundred forty-five male and 116 female patients met inclusion criteria. Correlated against 24PWT, GPP has the strongest association with a correlation of 0.80 for males and 0.88 for females. PPD has a weaker correlation of 0.64 for males and 0.61 for females (R2 = 0.38 overall). Factor analysis identified two components associated with incontinence. A "leakage" component correlated best with 24PWT and GPP. Additionally, an "age" component implies that despite stable 24PWT values, older patients increase GPP while PPD decreases. CONCLUSIONS: Self-reported PPD is an unreliable measure of incontinence as this variable only measures 38% of the variation of urinary incontinence volume. Patients at a given PPD level present with a wide range of 24PWT values. Older patients have higher per-pad leakage. Future incontinence studies should report 24PWT to ensure the most reliable and uniform data. PMID- 17080416 TI - Analysis of outcomes of single polypropylene mesh in total pelvic floor reconstruction. AB - AIMS: A 2.5-year outcome analysis was performed on patients who underwent transvaginal repair of total pelvic organ prolapse with single polypropylene mesh. A description of the repair technique using a tension-free 4-point fixation is also reviewed. METHODS: After proper vaginal dissection, a specially fashioned "H" shaped polypropylene mesh is positioned and fixed at 4-points. With a single piece of mesh, the anterior arms provide mid-urethral and bladder neck support, the mid-portion of the mesh corrects anterior compartment defects, and the posterior arms aid in vaginal vault suspension. Initially, bone anchors were utilized for anterior fixation, but currently a tension-free method is used. A retrospective analysis using chart review was performed on 96 patients who underwent this procedure from January 2000 to June 2005. Additional information was gathered by a telephone survey using a questionnaire. Statistical analysis was performed using Student's t-test, with Sigma Stat(R). RESULTS: Seventy-six patients (79%) were available with a mean follow-up time of 30.7 +/- 1.7 months and mean age of 69.3 +/- 11.3. Among those with follow-up, 36 patients (47.4%) underwent concurrent hysterectomies. Recurrence of prolapse was reported by four patients (5.2%). Sixty-eight patients (89%) were completely dry or almost dry, defined as an occasional leak. For those with preoperative incontinence (n = 36), average pad use per day decreased significantly from 2.1 +/- 0.4 to 0.8 +/- 0.2 (P < 0.005) postoperatively. Twelve patients (15.7%) reported of de novo urgency. Six patients required reoperation including excision of vaginal mesh erosion (2), uretholysis for obstruction (1), removal of palpable vaginal suture (1), and recurrent SUI (2). Among the 21 patients who are sexually active, 19 denied any dyspareunia (90.4%). Patient satisfaction was high, as the mean value was 7.9 +/- 0.3 on a scale of 1 (least satisfied) to 10 (most satisfied). CONCLUSIONS: Transvaginal repair of complete pelvic prolapse using polypropylene mesh is a safe and efficacious option, with minimal recurrence of prolapse and SUI. While two patients had vaginal erosions, no urethral or bladder erosions occurred. Patient satisfaction was overall favorable. PMID- 17080417 TI - Update on the neurology of Parkinson's disease. AB - The differential diagnosis of a patient with apparent Parkinson's Disease (PD) and bladder symptoms is considered and the bladder dysfunction of Multiple System Atrophy (MSA) is reviewed. Recent insights into the progression of the neuropathology of PD have enabled thinking about the stage of the disease at which bladder dysfunction is likely to occur and the expected clinical context of the problem. Bladder symptoms of neurological origin are likely in a patient who has had treated motor symptoms for some years and in whom the ongoing neuropathology has progressed beyond involvement of the basal ganglia, so that symptoms due to cortical dysfunction as well as the adverse effects of dopaminergic medication are also confounding factors. Bladder symptoms in a man with lesser neurological disability should be investigated to exclude underlying outflow obstruction. Possible management options are considered. PMID- 17080419 TI - Effect of moisture and pressure on tablet compaction studied with FTIR spectroscopic imaging. AB - FTIR spectroscopic imaging using a diamond ATR accessory has been applied to examine the influence of moisture and compression pressure on the density and components distribution of compacted pharmaceutical tablets. The model drug and excipient used within this study are ibuprofen and hydroxypropylmethylcellulose (HPMC). Chemical images of these compacted tablets were captured in situ without removing the tablet between measurements. A powder mixture of both, drug and excipient, prior to compaction, were subjected to a controlled environment, using a controlled humidity cell. Histograms were plotted to assess the density distribution quantitatively. This FTIR spectroscopic imaging approach enabled both measurement of water sorption and enhanced visualization of the density distribution of the compacted tablets. PMID- 17080420 TI - Under the microscope: single molecule symposium at the University of Michigan, 2006. AB - In recent years, a revolution has occurred in the basic sciences, which exploits novel single molecule detection and manipulation tools to track and analyze biopolymers in unprecedented detail. A recent Gordon Research Conference style meeting, hosted by the University of Michigan, highlighted current status and future perspectives of this rising field as researchers begin to integrate it with mainstream biology and nanotechnology. PMID- 17080418 TI - Molecular dynamics simulations of RNA: an in silico single molecule approach. AB - RNA molecules are now known to be involved in the processing of genetic information at all levels, taking on a wide variety of central roles in the cell. Understanding how RNA molecules carry out their biological functions will require an understanding of structure and dynamics at the atomistic level, which can be significantly improved by combining computational simulation with experiment. This review provides a critical survey of the state of molecular dynamics (MD) simulations of RNA, including a discussion of important current limitations of the technique and examples of its successful application. Several types of simulations are discussed in detail, including those of structured RNA molecules and their interactions with the surrounding solvent and ions, catalytic RNAs, and RNA-small molecule and RNA-protein complexes. Increased cooperation between theorists and experimentalists will allow expanded judicious use of MD simulations to complement conceptually related single molecule experiments. Such cooperation will open the door to a fundamental understanding of the structure function relationships in diverse and complex RNA molecules. . PMID- 17080421 TI - Mechanisms of DNA binding determined in optical tweezers experiments. AB - The last decade has seen rapid development in single molecule manipulation of RNA and DNA. Measuring the response force for a particular manipulation has allowed the free energies of various nucleic acid structures and configurations to be determined. Optical tweezers represent a class of single molecule experiments that allows the energies and structural dynamics of DNA to be probed up to and beyond the transition from the double helix to its melted single strands. These experiments are capable of high force resolution over a wide dynamic range. Additionally, these investigations may be compared with results obtained when the nucleic acids are in the presence of proteins or other binding ligands. These ligands may bind into the major or minor groove of the double helix, intercalate between bases or associate with an already melted single strand of DNA. By varying solution conditions and the pulling dynamics, energetic and dynamic information may be deduced about the mechanisms of binding to nucleic acids, providing insight into the function of proteins and the utility of drug treatments. PMID- 17080422 TI - Analysis of the inter-molecular interactions between amino acids and acetone by THz spectroscopy. AB - The THz spectra of amino acids after application of spots of acetone were measured. The 0.6 THz band was commonly observed in many amino acids that formed the intra-molecular salt structure. The band can be attributed to the interaction vibration from the common structural configuration of amino acids and acetone molecules. The evidence suggests that the vibration between the amino acids with intra-molecular salt structure and acetone has a peak at 0.6 THz. A model of the interaction vibration of acetone and the functional groups of amino acids is proposed. PMID- 17080423 TI - Analysis, interpretation, and extrapolation of dermal permeation data using diffusion-based mathematical models. AB - New dermal penetration data have been measured in both "infinite" and finite dose experiments on a range of compounds of varying lipophilicities. The data are analyzed, using parameter fitting, to determine the values of parameters governing the overall skin absorption processes. Two one-dimensional diffusion models are used. The first is novel, and well suited to the modeling of dermal uptake in occupational exposure scenarios. The second is an implementation of a model taken from the literature. The models are compared in a variety of exposure scenarios, and exhibit good mutual agreement. Both successfully reproduce expected features of the absorption process. Penetration parameters are determined by analyzing both infinite and finite dose data. Prediction of dermal absorption with finite dose scenarios is carried out and compared with experimental data obtained under these conditions. Parameters determined may also have an important role in improving the reliability of predictive QSARs used to estimate the extent of penetration of untested molecules. PMID- 17080424 TI - Quantitation of aggregate levels in a recombinant humanized monoclonal antibody formulation by size-exclusion chromatography, asymmetrical flow field flow fractionation, and sedimentation velocity. AB - Size-exclusion high-performance liquid chromatography (SE-HPLC, SEC) is the long standing biopharmaceutical industry standard for quantitation of soluble protein aggregates. Recently, sedimentation velocity analytical ultracentrifugation (SV AUC) has emerged as a possible orthogonal technique to SEC for soluble aggregate quantitation. Moreover, asymmetrical flow field flow fractionation (AF4) has shown early promise in quantifying protein aggregates, both soluble and insoluble. We report soluble aggreg ate quantities measured by SEC, AF4, and SV AUC analyzed by SEDFIT/c(s) for acid stressed and unstressed samples of a recombinant humanized monoclonal antibody. In equivalent antibody samples, SV AUC, and AF4 detect markedly higher total aggregate levels than SEC. Furthermore, SEC fails to detect higher molecular weight soluble aggregates apparent in SV-AUC and AF4 analyses. Pooled fractions containing soluble dimeric aggregates were purified and re-analyzed by both SV-AUC and SEC. Reinjection of purified dimer onto the SEC column induces formation of detectable quantities of monomer and trimer. All sample types show statistically significant (p-values<0.01) antibody losses through the SEC column. This incomplete mass recovery from SEC indicates probable antibody physical adsorption to gel filtration media. Analysis of the sedimentation behavior of high molecular weight components suggests increased molecular asphericity with increasing molecular weight. We present an aggregation model based on nearly linear end-to-end assembly of monomeric subunits which is shown to be consistent with SV-AUC, SEC, AF4, and dynamic light scattering (DLS) results. PMID- 17080425 TI - Iontophoretic drug delivery across human nail. AB - Topical trans-nail delivery of antifungal drugs is limited by several physicochemical and physiological factors. Use of chemical permeation enhancers has been a common approach for enhancing trans-nail delivery of drugs. The potential of physical permeation enhancement techniques has been found to be higher than the potential of chemical permeation enhancers in transdermal delivery of hydrophilic drugs and macromolecular therapeutic agents. However, application of physical permeation enhancement techniques has not been explored for trans-nail drug delivery. In the current work, iontophoresis was applied across human nail in vitro to assess its efficiency in enhancing drug delivery. Salicylic acid (SA) was used as test diffusant. The influence of pH, ionic strength, and current density was studied. Obviously, increase in current density increased the trans-nail transport flux. It appears that about 50-100 mM ionic strength is required for optimal conduction of electric current across nail. The flux enhancement factor (iontophoretic flux/passive flux) also increased with increase in pH due to increased ionization of SA. This study demonstrates the efficacy of iontophoresis in enhancing the trans-nail delivery of drugs. PMID- 17080426 TI - Air-liquid interface (ALI) culture of human bronchial epithelial cell monolayers as an in vitro model for airway drug transport studies. AB - Serially passaged normal human bronchial epithelial (NHBE) cell monolayers were established on Transwell inserts via an air-liquid interface (ALI) culture method. NHBE cells were seeded on polyester Transwell inserts, followed by an ALI culture from day 3, which resulted in peak TEER value of 766+/-154 Omegaxcm2 on the 8th day. Morphological characteristics were observed by light microscopy and SEM, while the formation of tight junctions was visualized by actin staining, and confirmed successful formation of a tight monolayer. The transepithelial permeability (Papp) of model drugs significantly increased with the increase of lipophilicity and showed a good linear relationship, which indicated that lipophilicity is an important factor in determining the Papp value. The expression of P-gp transporter in NHBE cell monolayers was confirmed by the significantly higher basolateral to apical permeability of rhodamine123 than that of reverse direction and RT-PCR of MDR1 mRNA. However, the symmetric transport of fexofenadine.HCl in this NHBE cell monolayers study seems to be due to the low expression of P-gp transporter and/or to its saturation with high concentration of fexofenadine.HCl. Thus, the development of tight junction and the expression of P-gp in the NHBE cell monolayers in this study imply that they could be a suitable in vitro model for evaluation of systemic drug absorption via airway delivery, and that they reflect in vivo condition better than P-gp over-expressed cell line models. PMID- 17080427 TI - The relationship between water vapor absorption and desorption by phospholipids and bilayer phase transitions. AB - Water vapor absorption and desorption at 25 degrees C and phase transition temperatures of phospholipid bilayers were measured as a function of relative humidity (RH) to better understand how the patterns of water vapor absorption and desorption are linked to corresponding phase changes induced by the level of hydration. Comparisons were made of the dipalmitoyl and palmitoyloleyol esters of glycerol derivatized with phosphatidyl-choline, -glycerol, -ethanolamine and with phosphatidic acid. The results suggest that the extent of water vapor absorption and desorption at a given RH reflects the combined effects of water-polar group interaction and access of water to the polar region as controlled by intra- and interbilayer molecular packing and intermolecular attractive and repulsive interactions. The results further suggest that the extent of water vapor absorption and desorption over a range of relative humidities reflects the combined effects of the polar group's ability to interact with water, the access that water has to the polar groups as determined by molecular size and various intermolecular and intrabilayer forces of attraction and repulsion, and interbilayer interactions which influence the degree of order/disorder present in the overall solid-state structure. This behavior is also reflected in the changes observed in the various bilayer phase transition temperatures as a function of RH. Analyses of absorption isotherms suggests that after exceeding a critical RH, water initially interacting with these phospholipids most likely forms either stoichiometric or nonstoichiometric crystal hydrates, as with the disaturated derivatives, or hydrated mesophases, as with the gel states of the monounsaturated derivatives. PMID- 17080428 TI - Notch1 and Notch2 receptors influence progressive hair graying in a dose dependent manner. AB - The Notch signaling pathway is involved in diverse biological processes such as cell fate decisions or stem cell maintenance. In this study, we assessed the role of this pathway for melanocyte development and hair pigmentation using RBP Jkappa, Notch1, and Notch2 conditional knockout mice. Disruption of the Notch pathway by inactivating RBP-Jkappa in the melanocyte lineage using Tyr::Cre mice led to a severe coat color dilution. Similarly, hair graying was observed when Notch1 and/or Notch2 receptors were ablated in melanocytes. This phenotype was proportional to the number of floxed Notch alleles, with the most pronounced effect seen in Tyr::Cre/degrees; Notch1(flox/flox); Notch2(flox/flox) mice. Deletion of Notch1 and/or Notch2 in melanoblasts did not induce a congenital defect. The number of Dct-expressing cells at embryonic stages was not affected, but melanocytes located within the hair matrix progressively disappeared during the first regeneration of the hair follicle. In contrast, non-follicular melanocytes and pigmentation in the dermis and in the choroid were not affected. We suggest that both Notch1 and Notch2 receptors contribute to the maintenance of melanoblasts and melanocyte stem cells, and are essential for proper hair pigmentation. PMID- 17080429 TI - Beneficial effects of creatine, CoQ10, and lipoic acid in mitochondrial disorders. AB - Mitochondrial disorders share common cellular consequences: (1) decreased ATP production; (2) increased reliance on alternative anaerobic energy sources; and (3) increased production of reactive oxygen species. The purpose of the present study was to determine the effect of a combination therapy (creatine monohydrate, coenzyme Q(10), and lipoic acid to target the above-mentioned cellular consequences) on several outcome variables using a randomized, double-blind, placebo-controlled, crossover study design in patients with mitochondrial cytopathies. Three patients had mitochondrial encephalopathy, lactic acidosis, and stroke-like episodes (MELAS), four had mitochondrial DNA deletions (three patients with chronic progressive external ophthalmoplegia and one with Kearns Sayre syndrome), and nine had a variety of other mitochondrial diseases not falling into the two former groups. The combination therapy resulted in lower resting plasma lactate and urinary 8-isoprostanes, as well as attenuation of the decline in peak ankle dorsiflexion strength in all patient groups, whereas higher fat-free mass was observed only in the MELAS group. Together, these results suggest that combination therapies targeting multiple final common pathways of mitochondrial dysfunction favorably influence surrogate markers of cellular energy dysfunction. Future studies with larger sample sizes in relatively homogeneous groups will be required to determine whether such combination therapies influence function and quality of life. PMID- 17080430 TI - Prolonged residual catatonia or prolonged neuroleptic-induced Parkinsonism following catatonic stupor in a patient with manic-depressive illness. PMID- 17080431 TI - Combining optical tweezers and scanning probe microscopy to study DNA-protein interactions. AB - We present the first results obtained with a new instrument designed and built to study DNA-protein interactions at the single molecule level. This microscope combines optical tweezers with scanning probe microscopy and allows us to locate DNA-binding proteins on a single suspended DNA molecule. A single DNA molecule is stretched taut using the optical tweezers, while a probe is scanned along the molecule. Interaction forces between the probe and the sample are measured with the optical tweezers. The instrument thus enables us to correlate mechanical and functional properties of bound proteins with the tension within the DNA molecule. The typical friction force between a micropipette used as probe and a naked DNA molecule was found to be <1 pN. A 16 micro m DNA molecule with approximately 10 15 digoxygenin (DIG) molecules located over a 90 nm range in the middle of the DNA was used as a model system. By scanning with an antidigoxygenin (alpha-DIG) antibody-coated pipette we were able to localize these sites by exploiting the high binding affinity between this antibody-antigen pair. The estimated experimental resolution assuming an infinitesimally thin and rigid probe and a single alpha-DIG/DIG bond was 15 nm. PMID- 17080432 TI - FRET and colocalization analyzer--a method to validate measurements of sensitized emission FRET acquired by confocal microscopy and available as an ImageJ Plug-in. AB - Fluorescence resonance energy transfer (FRET) between an adequate pair of fluorophores is an indication of closer proximity than colocalization and is used by biologists to study fluorescently modified protein interactions inside cells. We present a method for visualization of FRET images acquired by confocal sensitized emission, involving excitation of the donor fluorophore and detection of the energy transfer as an emission from the acceptor fluorophore into the FRET channel. Authentic FRET signal measurements require the correction from the FRET channel of the undesired bleed-through signals (BT) resulting from both the leak through of the donor emission and the direct acceptor emission. Our method reduces the interference of the user to a minimum by analyzing the entire image, pixel by pixel. It proposes imaging treatments and the display of control images to validate the BT calculation and the image corrections. It displays FRET images as a function of the colocalization of the two fluorescent partners. Finally, it proposes an alternative to normalization of the FRET intensities to compare FRET signal variations between samples. This method called "FRET and Colocalization Analyzer" has been implemented in a Plug-in of the freely available ImageJ software. It is particularly adapted when transient expression of the fluorescent proteins is used thereby giving very variable expression levels or when the colocalization of the two partners is varying in proportion, in amount, and in size, as a function of time. The method and program are validated using the analysis of the spatio-temporal interactions between a G-protein coupled receptor, the tachykinin NK2 receptor, and the beta-arrestin 2 as an example. PMID- 17080433 TI - Smoking-responsive juvenile-onset Parkinsonism. AB - We describe a patient with juvenile levodopa-responsive Parkinsonism who reported a dramatic response to cigarette smoking with transient but marked improvement of motor symptoms associated with oculogyric crises and psychotic behavior. His beta CIT single-photon emission computed tomography scan showed a complete absence of presynaptic dopaminergic nerve terminals. PMID- 17080434 TI - Differentiation syndrome with lethal lung injury after arsenic trioxide treatment for myelodysplastic syndrome. PMID- 17080435 TI - Detecting functional nodes in large-scale cortical networks with functional magnetic resonance imaging: a principal component analysis of the human visual system. AB - This study aimed to demonstrate how a regional variant of principal component analysis (PCA) can be used to delineate the known functional subdivisions of the human visual system. Unlike conventional eigenimage analysis, PCA was carried out as a second-level analysis subsequent to model-based General Linear Model (GLM) type functional activation mapping. Functional homogeneity of the functional magnetic resonance imaging (fMRI) time series within and between clusters was examined on several levels of the visual network, starting from the level of individual clusters up to the network level comprising two or more distinct visual regions. On each level, the number of significant components was identified and compared with the number of clusters in the data set. Eigenimages were used to examine the regional distribution of the extracted components. It was shown that voxels within individual clusters and voxels located in bilateral homologue visual regions can be represented by a single component, constituting the characteristic functional specialization of the cluster(s). If, however, PCA was applied to time series of voxels located in functionally distinct visual regions, more than one component was observed with each component being dominated by voxels in one of the investigated regions. The model of functional connections derived by PCA was in accordance with the well-known functional anatomy and anatomical connectivity of the visual system. PCA in combination with conventional activation mapping might therefore be used to identify the number of functionally distinct nodes in an fMRI data set in order to generate a model of functional connectivity within a neuroanatomical network. PMID- 17080436 TI - A history of human-like dieting alters serotonergic control of feeding and neurochemical balance in a rat model of binge-eating. AB - OBJECTIVE: This study replicated a model of stress-induced binge-eating in rats with a history of caloric restriction (HCR), tested their response to SSRI (fluoxetine) treatment, and explored changes in brain monoamine levels. METHOD: Young female rats with no-HCR/no-Stress, no-HCR/Stress, HCR/no-Stress, and HCR+Stress (binge-eating) were treated with fluoxetine. Post-mortem levels of serotonin, dopamine, and metabolites were assessed from brain regions key to feeding and reward. RESULTS: A 3 mg/kg dose of fluoxetine without effect in the no-HCR groups suppressed intake of HCR groups, normalizing the binge-eating of HCR/Stress rats. No differences in monoamines were detected in the hypothalamus or tegmentum but a strong positive relationship between accumbens serotonin and dopamine turnover in no-HCR rats was absent in rats with HCR. CONCLUSION: Despite lack of hunger, a history of human-like dieting alters serotonin function in ways suggesting consequences not only to feeding but also control of reward and mood that are dependent on dopamine/serotonin interactions. PMID- 17080437 TI - Relationship between regional hemodynamic activity and simultaneously recorded EEG-theta associated with mental arithmetic-induced workload. AB - Theta increases with workload and is associated with numerous processes including working memory, problem solving, encoding, or self monitoring. These processes, in turn, involve numerous structures of the brain. However, the relationship between regional brain activity and the occurrence of theta remains unclear. In the present study, simultaneous EEG-fMRI recordings were used to investigate the functional topography of theta. EEG-theta was enhanced by mental arithmetic induced workload. For the EEG-constrained fMRI analysis, theta-reference time series were extracted from the EEG, reflecting the strength of theta occurrence during the time course of the experiment. Theta occurrence was mainly associated with activation of the insular cortex, hippocampus, superior temporal areas, cingulate cortex, superior parietal, and frontal areas. Though observation of temporal and insular activation is in accord with the theory that theta specifically reflects encoding processes, the involvement of several other brain regions implies that surface-recorded theta represents comprehensive functional brain states rather than specific processes in the brain. The results provide further evidence for the concept that emergent theta band oscillations represent dynamic functional binding of widely distributed cortical assemblies, essential for cognitive processing. This binding process may form the source of surface recorded EEG theta. PMID- 17080438 TI - Effect of corpus callosum damage on ipsilateral motor activation in patients with multiple sclerosis: a functional and anatomical study. AB - Functional MRI (fMRI) studies have shown increased activation of ipsilateral motor areas during hand movement in patients with multiple sclerosis (MS). We hypothesized that these changes could be due to disruption of transcallosal inhibitory pathways. We studied 18 patients with relapsing-remitting MS. Conventional T1- and T2-weighted images were acquired and lesion load (LL) measured. Diffusion tensor imaging (DTI) was performed to estimate fractional anisotropy (FA) and mean diffusivity (MD) in the body of the corpus callosum (CC). fMRI was obtained during a right-hand motor task. Patients were studied to evaluate transcallosal inhibition (TCI, latency and duration) and central conduction time (CCT). Eighteen normal subjects were studied with the same techniques. Patients showed increased MD (P < 0.0005) and reduced FA (P < 0.0005) in the body of the CC. Mean latency and duration of TCI were altered in 12 patients and absent in the others. Between-group analysis showed greater activation in patients in bilateral premotor, primary motor (M1), and middle cingulate cortices and in the ipsilateral supplementary motor area, insula, and thalamus. A multivariate analysis between activation patterns, structural MRI, and neurophysiological findings demonstrated positive correlations between T1-LL, MD in the body of CC, and activation of the ipsilateral motor cortex (iM1) in patients. Duration of TCI was negatively correlated with activation in the iM1. Our data suggest that functional changes in iM1 in patients with MS during a motor task partially represents a consequence of loss of transcallosal inhibitory fibers. PMID- 17080439 TI - Event-related functional near-infrared spectroscopy (fNIRS) based on craniocerebral correlations: reproducibility of activation? AB - The purpose of the present study was to assess the retest reliability of cortical activation detected by event-related functional near-infrared spectroscopy (fNIRS) based on craniocerebral correlations. Isolated functional activation was evoked in the motor cortex by a periodically performed finger-tapping task. During 44-channel fNIRS recording, 12 subjects performed 30 trials of right and left index finger tapping in two sessions. The retest interval was set to 3 weeks. Simple correlations of the contrast t-values supplemented by scatterplots, channel-wise intraclass correlation coefficients (ICC), as well as reproducibility indices for the size and the location of the detected activation were calculated. The results at the group level showed sufficient single measure ICCs (up to 0.80) and excellent reproducibility of the size and the location (up to 89% were reproducible). Comparisons of the intersession group amplitudes demonstrate that the fNIRS signals were stable across time in a retest study design: the number of significant differences was less than randomly occurring false-positive activated channels if an alpha level of 5% is chosen. Effect size analyses indicated that the intersession amplitude differences are small (mean < 0.25). For deoxyhemoglobin and oxyhemoglobin distinct statistical power profiles were revealed regarding the activation vs. baseline contrast as well as the intersession amplitude differences, indicating a higher sensitivity of deoxyhemoglobin for local hemodynamic changes. The results suggest that sensorimotor activation assessed by event-related fNIRS based on craniocerebral correlations is sufficiently reproducible at the group level. PMID- 17080440 TI - Assessing the influence of scanner background noise on auditory processing. I. An fMRI study comparing three experimental designs with varying degrees of scanner noise. AB - We compared two experimental designs aimed at minimizing the influence of scanner background noise (SBN) on functional MRI (fMRI) of auditory processes with one conventional fMRI design. Ten subjects listened to a series of four one-syllable words and had to decide whether two of the words were identical. This was contrasted with a no-stimulus control condition. All three experimental designs had a duration of approximately 17 min: 1) a behavior interleaved gradients (BIG; Eden et al. [1999] J Magn Reson Imaging 41:13-20) design (repetition time, TR, = 6 s), where stimuli were presented during the SBN-free periods between clustered volume acquisitions (CVA); 2) a sparse temporal sampling technique (STsamp; e.g., Gaab et al., [2003] Neuroimage 19:1417-1426) acquiring only one set of slices following each of the stimulations with a 16-s TR and jittered delay times between stimulus offset and image acquisition; and 3) an event-related design with continuous scanning (ERcont) using the stimulation design of STsamp but with a 2-s TR. The results demonstrated increased signal within Heschl's gyrus for the STsamp and BIG-CVA design in comparison to ERcont as well as differences in the overall functional anatomy among the designs. The possibility to obtain a time course of activation as well as the full recovery of the stimulus- and SBN induced hemodynamic response function signal and lack of signal suppression from SBN during the STsamp design makes this technique a powerful approach for conducting auditory experiments using fMRI. Practical strengths and limitations of the three auditory acquisition paradigms are discussed. PMID- 17080441 TI - Task demand modulation of steady-state functional connectivity to primary motor cortex. AB - Correlations in blood oxygen level-dependent (BOLD) MRI signals from separate areas within the human brain have been used as a measure of functional connectivity. Steady-state measures of interregional correlations are particularly useful because they do not depend on the specific design of a task nor on subtracting conditions in a blocked design task. However, the conditions under which such correlations are measured may influence these indices of functional connectivity. The aim of this study was to investigate the influence of task demand on interregional correlations within the motor system. Specifically, tapping rates in audibly paced finger-tapping tasks were controlled and varied between runs in order to observe their effects on interregional correlations to contralateral primary motor cortex (PM). Regions of interest included the supplementary motor area, ipsilateral cerebellum, ipsilateral auditory cortex, and a control region. It was found that tapping rate was a significant factor in determining the mean correlation of some regions to PM, and that correlations measured during tapping in general increased relative to resting state. Furthermore, analysis of the percent of voxels in each region significantly correlated to PM suggested that changes in the mean correlation of that region to PM could be accounted for by changes in the fraction of significantly correlated voxels within a region. This provides insight into the manner in which steady-state correlations are modified in response to different task demands and further evidence that low-frequency fluctuations in BOLD signals reflect functional connectivity. PMID- 17080442 TI - Methylphenidate-induced activation of the anterior cingulate but not the striatum: a [15O]H2O PET study in healthy volunteers. AB - The dopaminergic system has been implicated in the pathogenesis and treatment of a variety of neuropsychiatric disorders, such as schizophrenia, depression, and addiction. (Dys)function of the dopaminergic system may be studied by combining [15O]H2O PET with a dopaminergic drug challenge. In this pilot study we investigated the suitability of the dopamine reuptake blocker methylphenidate (MP) as a dopaminergic probe. Measurements of regional cerebral blood flow (rCBF) were made at 10 and 30 min after placebo and MP (0.25 mg/kg) injection to seven healthy volunteers. During scanning the behavioral condition of the subjects was standardized using a continuous performance task. Growth hormone levels were assessed and subjective ratings were obtained. MP significantly elevated growth hormone levels. After receiving MP, the subjective experience varied from neutral to highly pleasurable. Ten minutes after MP administration, significant relative increases in rCBF were found in the rostral anterior cingulate (AC), temporal poles, and the supplementary motor area. Significant reductions were seen in the superior temporal gyri, right medial frontal gyrus, and right inferior parietal cortex. At 30 min after MP administration, increases were seen in the AC, temporal pole, and right cerebellum. No changes were observed in the striatum. The activation in the right rostral AC was significantly higher in the subjects with the highest euphoria scores compared to the subjects with minimal MP-induced changes in euphoria. We suggest that the combined MP challenge with functional imaging, as described in our study, may be a useful tool to study the functional integrity of the dopaminergic system in psychiatric disorders. PMID- 17080443 TI - Subthalamic nucleus stimulation is efficacious in patients with Parkinsonism and LRRK2 mutations. AB - Stimulation of the subthalamic nucleus (STN) improves motor signs in patients with levodopa-responsive Parkinson's disease (PD). Mutations in the leucine-rich repeat kinase-2 (LRRK2) gene cause Parkinsonism. We assessed 69 patients under STN stimulation and found heterozygous LRRK2 mutations in 9 (G2019S in 8 and T2031S in 1). The age at onset of PD, the clinical characteristics before or after neurosurgery, and the clinical response to STN stimulation were similar in both groups. Two patients with the G2019S LRRK2 mutation still benefited from STN stimulation, 9 and 10 years after surgery. Patients with LRRK2 mutations are, therefore, good candidates for STN stimulation. PMID- 17080444 TI - Neurodegenerative overlap syndrome: Parkinsonism and motor neuron disorder. PMID- 17080445 TI - Trade names in publications. PMID- 17080446 TI - Narcissism and narcissistic defences in the eating disorders. AB - OBJECTIVE: This study examined the associations between eating pathology and narcissism in an eating-disordered group. Narcissism was conceptualized in terms of both its core element (entitlement, grandiosity) and the narcissistic defenses that are used to maintain self-esteem. METHOD: Seventy non-clinical and 84 eating disordered patients completed a measure of the different elements of narcissism, and a standardized measure of eating pathology. RESULTS: The eating-disordered group scored higher than the non-clinical women on the measures of core narcissism and of the narcissistically abused style ("poor me" defense). The pattern of dimensional associations between narcissism and eating pathology was highly similar across the clinical and nonclinical groups, with the narcissistic defenses playing the strongest role. The poisonous pedagogy style ("bad you" defense) was positively associated with restrictive attitudes toward eating, while the narcissistically abused style was positively associated with restraint, eating concern, body shape concern, and body weight concern. CONCLUSION: The narcissistic defenses are particularly relevant in understanding the eating disorders. Implications for future research are outlined, and suggestions are made about the need to assess and respond to these associations in treatment. PMID- 17080447 TI - Maintenance of internet-based prevention: a randomized controlled trial. AB - OBJECTIVE: Excessive weight or shape concerns and dieting are among the most important and well-established risk factors for the development of symptoms of disordered eating or full-syndrome eating disorders. Prevention programs should therefore target these factors in order to reduce the likelihood of developing an eating disorder. The aims of this study were to determine the short-term and maintenance effects of an internet-based prevention program for eating disorders. METHOD: One hundred female students at two German universities were randomly assigned to either an 8-week intervention or a waiting-list control condition and assessed at preintervention, postintervention, and 3-month follow-up. RESULTS: Compared with the control group, the intervention produced significant and sustained effects for high-risk women. CONCLUSION: Internet-based prevention is effective and can be successfully adapted to a different culture. PMID- 17080448 TI - Treatment of bulimia nervosa: where are we and where are we going? AB - OBJECTIVE: The purpose of this article is to review the extant treatment literature on bulimia nervosa and to offer suggestions for future research directions. METHOD: The available treatment studies regarding both pharmacotherapy and psychotherapy are reviewed. RESULTS: Both pharmacotherapy and psychotherapy appear to play a role in the treatment of bulimia nervosa; however, available data suggest that cognitive behavioral therapy remains the treatment of choice. CONCLUSION: Additional work is clearly indicated regarding assisted and unassisted self-help. An enhanced form of CBT and the integrative cognitive affective therapy both deserve further study. New approaches need to be piloted. More research is needed on treatment modeling. PMID- 17080449 TI - Cross-ethnic differences in eating disorder symptoms and related distress. AB - OBJECTIVE: This study investigated ethnic differences in the frequency of eating disorder symptoms and related distress in a U.S. college-based eating disorders screening program. METHOD: Responses to self-report questions and counselors' assessment were analyzed in an ethnically diverse sample (n = 5,435). RESULTS: The frequency of binge-eating, restrictive eating, vomiting, and amenorrhea did not differ significantly across ethnic groups. However, significant between-group differences were found with respect to modes of purging. Binge correlates (e.g., eating until uncomfortably full) were significantly more frequent among Caucasian than African American participants (p < .001). Binge eating was the best predictor of distress among Caucasians, African Americans, and Latinos, whereas vomiting was the best predictor of distress among Asians. Asian participants who used laxatives were significantly less likely to receive a recommendation for further evaluation than non-Asian participants. CONCLUSION: Ethnic diversity in symp tom prevalence and related distress was identified. Clinician recognition of this potential diversity may enhance culturally competent care for eating disorders. PMID- 17080450 TI - Predictors of weight maintenance after hospital discharge in adolescent anorexia nervosa. AB - OBJECTIVE: To analyze variables that predict weight maintenance in adolescent anorexia nervosa. METHOD: The Eating Attitudes Test, the Beck Depression Inventory, the Leyton Obsessional Inventory, the State and Trait Anxiety Inventory, and the Anorexia Nervosa Stages of Change Questionnaire were administered to 49 anorexia nervosa patients (mean age 14.3 years, SD 1.7) consecutively admitted to an eating disorder unit. They were evaluated at admission, at discharge, and after nine months follow-up. RESULTS: At discharge, patients had improved in body mass index (p < .001), eating attitudes (p = .002), depressive symptomatology (p = .001), and motivation to change (p < .001). Patients with good weight maintenance at follow-up had higher body mass index (p = .017) at admission, lower abnormal eating attitudes (p = .035), depressive symptomatology (p = .026), and higher motivation to change (p = .004) at discharge. Logistic regression analysis showed a high motivation to change at discharge and a high body mass index at admission to be predictors of weight maintenance. CONCLUSION: High motivation to change, low abnormal eating attitudes, depressive symptomatology at discharge, and high body mass index at admission are associated with weight maintenance in adolescent anorexia nervosa. PMID- 17080451 TI - A comparison of energy intake and food selection during laboratory binge eating episodes in obese women with and without a binge eating disorder diagnosis. AB - OBJECTIVE: The purpose of the study was to compare energy intake and food selection of laboratory binge eating episodes in obese women with and without binge eating disorder (BED). METHOD: Twenty women, 12 meeting BED criteria, and 8 BMI and age matched obese controls, engaged in a laboratory binge eating episode. RESULTS: BED participants consumed significantly more total food in kilocalories than the non-BED obese group and more kilocalories of fat. However, there were no differences between the groups in the proportion of calories from any macronutrient. There was a nonsignificant trend for the BED group to consume more total grams of food. During the binge, the BED group consumed more dairy products. CONCLUSION: Results of the study confirmed that women with BED consumed significantly more total food in kilocalories than the non-BED obese women when they were allowed to have a binge eating episode in a laboratory setting. PMID- 17080452 TI - Change of nitric oxide concentration in men exposed to a 1.5 T constant magnetic field. AB - This study was carried out in order to determine nitric oxide (NO) production immediately after a 1.5 T magnetic field 30 min exposure to an experimental group, comprising 33 healthy young male volunteers aged 18-26 years old. In addition, a control group, comprising 30 healthy male volunteers aged 19-26 years old, was not exposed to the magnetic field and their NO levels were also measured. The experimental group was exposed using a magnetic resonance imaging (MRI) apparatus. Nitrite and nitrate concentrations were determined by UV-VIS spectrophotometer. The results, related to the parameters measured in this study, were analyzed by one-way ANOVA. Total nitrite concentration in post-magnetic field samples was found to be higher than in pre-magnetic field samples (P < .05). PMID- 17080453 TI - Measurements of skin surface temperature during mobile phone use. AB - We measured maximum temperature rises on the side of the face after 6 min of continuous mobile phone operation using two models of AMPS analog phones operating in the 835 MHz band and three early model GSM digital phones operating in the 900 MHz band. For the GSM phones the highest recorded temperature rise difference was 2.3 degrees C and for the AMPS phones it was 4.5 degrees C, both at locations on the cheek. The higher differential temperature rise between AMPS and GSM may reflect the higher maximum average operating power of AMPS (600 mW) versus GSM900 (250 mW). Additionally, we compared temperature changes at a consistent location on the cheek for an AMPS phone that was inoperative (-0.7 degrees C), transmitting at full power (+2.6 degrees C) and in stand-by mode (+2.0 degrees C). Our results suggest that direct RF heating of the skin only contributes a small part of the temperature rise and that most is due to heat conduction from the handset. PMID- 17080454 TI - Low-power millimeter wave radiations do not alter stress-sensitive gene expression of chaperone proteins. AB - This article reports experimental results on the influence of low-power millimeter wave (MMW) radiation at 60 GHz on a set of stress-sensitive gene expression of molecular chaperones, namely clusterin (CLU) and HSP70, in a human brain cell line. Selection of the exposure frequency is determined by its near future applications for the new broadband civil wireless communication systems including wireless local area networks (WLAN) for domestic and professional uses. Frequencies around 60 GHz are strongly attenuated in the earth's atmosphere and such radiations represent a new environmental factor. An exposure system operating in V-band (50-75 GHz) was developed for cell exposure. U-251 MG glial cell line was sham-exposed or exposed to MMW radiation for different durations (1 33 h) and two different power densities (5.4 microW/cm(2) or 0.54 mW/cm(2)). As gene expression is a multiple-step process, we analyzed chaperone proteins induction at different levels. First, using luciferase reporter gene, we investigated potential effect of MMWs on the activation of transcription factors (TFs) and gene promoter activity. Next, using RT-PCR and Western blot assays, we verified whether MMW exposure could alter RNA accumulation, translation, or protein stability. Experimental data demonstrated the absence of significant modifications in gene transcription, mRNA, and protein amount for the considered stress-sensitive genes for the exposure durations and power densities investigated. The main results of this study suggest that low-power 60 GHz radiation does not modify stress-sensitive gene expression of chaperone proteins. PMID- 17080455 TI - Insight into the catalytic mechanism of arginine deiminase: functional studies on the crucial sites. AB - Arginine deiminase (ADI) catalyzes the irreversible hydrolysis of arginine to citrulline and ammonia. It belongs to a newly classified superfamily of guanidino group-modifying enzymes. Located in the catalytic center of Mycoplasma hominis ADI, some crucial sites (Asp160, Glu212, His268, and Asp270) are highly conserved among these enzymes. Here, we constructed five ADI single mutants D160E, E212D, H268F, H268Y, and D270E, and three double mutants D160E/D270E, D160E/E212D, and E212D/D270E, aiming to evaluate the contributions of these crucial residues to the structure, stability, and enzymatic activity of ADI, and to elucidate their roles in the catalytic process of this family of enzymes. Tryptophan emission fluorescence and circular dichroism were used to analyze the different effects of mutagenesis on these conserved residues on the secondary and tertiary structures of ADI. Urea-induced unfolding and trypsin digestion were applied to measure their stabilities against denaturants and proteases, respectively. Additionally, the enzymatic activities of ADI and its mutants were measured. Here, we report that all the mutations have little effect on the native structure of ADI. However, the substitutions on these crucial sites still interfere with the stability of ADI to different degrees. As these mutations impair both the substrate binding and the substrate induced conformational changes of ADI to different extents, most of the mutants except D160E (preserves about 30% of the enzymatic activity of wild type) have totally lost the enzymatic activity in the hydrolysis of arginine and the inhibitory ability on the proliferation of mouse melanoma cells. PMID- 17080456 TI - Elevated sister chromatid exchange frequencies in dividing human peripheral blood lymphocytes exposed to 50 Hz magnetic fields. AB - The in vitro cytomolecular technique, sister chromatid exchange (SCE), was applied to test the clastogenic potentiality of extremely low frequency (ELF) electromagnetic fields (EMFs) on human peripheral blood lymphocytes (HPBLs). SCE frequencies were scored in dividing peripheral blood lymphocytes (PBLs) from six healthy male blood donors in two rounds of experiments, R1 and R2, to determine reproducibility. Lymphocyte cultures in the eight experiments conducted in each round were exposed to 50 Hz sinusoidal (continuous or pulsed) or square (continuous or pulsed) MFs at field strengths of 1 microT or 1 mT for 72 h. A significant increase in the number of SCEs/cell in the grouped experimental conditions compared to the controls was observed in both rounds. The highest SCE frequency in R1 was 10.03 for a square continuous field, and 10.39 for a square continuous field was the second highest frequency in R2. DNA crosslinking at the replication fork is proposed as a model which could explain the mechanistic link between ELF EMF exposure and increased SCE frequency. PMID- 17080457 TI - Investigation of electric current perception thresholds of different EHS groups. AB - An increasing number of persons with health symptoms of unclear origin take refuge in the hypothesis that they suffer from electromagnetic hypersensitivity (EHS). So far EHS is not an accepted diagnosis and there is no validated test to verify the proposed relationship between electromagnetic fields and symptoms. Groups reporting EHS are very heterogeneous but share a belief that they have an increased sensitivity to electromagnetic fields. It was studied to which extent a quantitative indicator for electrosensitivity, the electric current perception threshold, and its variability coefficient, depend on the recruitment strategy for self-declared hypersensitive persons. Individual electrosensitivity was investigated by provocation of the lower arms to directly coupled 50 Hz electric currents. Self-declared EHS persons were selected from members of a self aid group, from responders to a newspaper call, and from persons actively asking for investigations in their search for help. It turned out that quantitative electrosensitivity was quite different among the three groups. It is interesting that the members of the EHS self aid group exhibit a considerable overlap with general population sample. Pooled together it could be shown that hypersensitive persons as a group differ significantly from the general population sample, however with a pronounced overlap with the normal range. It can be concluded that EHS groups are very inhomogeneous and contain numerous persons with no increased ability to perceive low frequency electric or magnetic fields. This investigation shows the importance of the study design, in particular of the recruitment strategies of EHS persons for the final outcome. PMID- 17080458 TI - Extremely low frequency weak magnetic fields enhance resistance of NN tobacco plants to tobacco mosaic virus and elicit stress-related biochemical activities. AB - Increasing evidence has accumulated concerning the biological effects of extremely low frequency magnetic fields (ELF-MFs) in different plant models. In the present study, effects of ELF-MFs in tobacco plants reacting to tobacco mosaic virus (TMV) with a hypersensitive response (HR) were evaluated. Plants were exposed for 8 or 24 h (either before or after TMV inoculation) to a static MF, at either -17 or 13 microT, combined with a 10 Hz sinusoidal MF with different intensities (25.6 or 28.9 microT). The working variables were the area and number of hypersensitive lesions in leaves. Following ELF-MFs exposure, an increased resistance was detected, particularly after an 8-h treatment, as shown by the decrease in lesion area and number. Moreover, two enzyme activities involved in resistance mechanisms were analyzed: ornithine decarboxylase (ODC) and phenylalanine ammonia-lyase (PAL). Uninoculated leaves previously exposed to ELF-MFs in general showed a significant increase relative to controls in ODC and PAL activities, in particular for 13 microT static MF plus 28.9 microT, 10 Hz sinusoidal MF (24 h) treatment. In conclusion, ELF-MFs seem to influence the HR of tobacco to TMV, as shown by the increased resistance and changes in ODC and PAL activities, indicating the reliability of the present plant model in the study of bioelectromagnetic interactions. PMID- 17080459 TI - Feasibility of future epidemiological studies on possible health effects of mobile phone base stations. AB - The increasing deployment of mobile communication base stations led to an increasing demand for epidemiological studies on possible health effects of radio frequency emissions. The methodological challenges of such studies have been critically evaluated by a panel of scientists in the fields of radiofrequency engineering/dosimetry and epidemiology. Strengths and weaknesses of previous studies have been identified. Dosimetric concepts and crucial aspects in exposure assessment were evaluated in terms of epidemiological studies on different types of outcomes. We conclude that in principle base station epidemiological studies are feasible. However, the exposure contributions from all relevant radio frequency sources have to be taken into account. The applied exposure assessment method should be piloted and validated. Short to medium term effects on physiology or health related quality of life are best investigated by cohort studies. For long term effects, groups with a potential for high exposure need to first be identified; for immediate effect, human laboratory studies are the preferred approach. PMID- 17080460 TI - Effects of 50 Hz sinusoidal magnetic fields on Hsp27, Hsp70, Hsp90 expression in porcine aortic endothelial cells (PAEC). AB - The aim of the present study was to investigate the influence of 50 Hz sinusoidal magnetic field on Hsp27, Hsp70, and Hsp90 expression in a model of primary culture of porcine aortic endothelial cells (PAEC). We took into consideration the Hsp profile in terms of mRNA expression, protein expression and protein localization inside the cells. The choice of the cell system was motivated by the involvement of the endothelial cells in the onset of many diseases; moreover, only few reports describe the effects of extremely low frequency magnetic fields (ELF-MFs) on such cells. ELF-MF exposure induced an increase in the mRNA levels of the three proteins, which was statistically significant for Hsp70. On the contrary, we did not observe any influence on Hsp27, Hsp70, and Hsp90 protein levels. Analysis in situ by immunofluorescence revealed that ELF-MF exposure affected the cellular distribution of Hsp27; in particular a partial relocalization in the nucleus was observed. PMID- 17080461 TI - Microsurgical anatomy of the superior orbital fissure. AB - The microanatomy of the superior orbital fissure (SOF) was studied in 96 sides of cadaver specimens. The SOF is a narrow bony cleft that lies at the apex of the orbit between the greater and lesser wings of the sphenoid. Through this fissure, many important structures enter the orbit from the middle cranial fossa including the third, fourth, sixth cranial nerves, and the ophthalmic branch of the fifth nerve. In addition, the superior opthalmamic vein exits the orbit to drain into the cavernous sinus via the SOF. The fissure can be divided into three anatomical regions by the annulus of Zinn (common annular tendon): the lateral, central, and inferior regions. The lateral wall of the SOF can also be divided between the upper and lower segments, and the angle between them was measured to be 144.27 degrees +/- 20.03 degrees . Defining these regions is useful in describing the course and placement of the nerves and vasculature in the SOF. Managing lesions at the orbital apex requires an extensive knowledge of the cranial base and the intracranial and extracranial relationships of the anatomical structures coursing through the SOF. The goal of this study was to describe the microanatomy of the SOF region in detail and to provide a reference for surgical procedures involving the orbital apex. PMID- 17080462 TI - Statistical and conformational analysis of the electron density of protein side chains. AB - Protein side chains make most of the specific contacts between proteins and other molecules, and their conformational properties have been studied for many years. These properties have been analyzed primarily in the form of rotamer libraries, which cluster the observed conformations into groups and provide frequencies and average dihedral angles for these groups. In recent years, these libraries have improved with higher resolution structures and using various criteria such as high thermal factors to eliminate side chains that may be misplaced within the crystallographic model coordinates. Many of these side chains have highly non rotameric dihedral angles. The origin of side chains with high B-factors and/or with non-rotameric dihedral angles is of interest in the determination of protein structures and in assessing the prediction of side chain conformations. In this paper, using a statistical analysis of the electron density of a large set of proteins, it is shown that: (1) most non-rotameric side chains have low electron density compared to rotameric side chains; (2) up to 15% of chi1 non-rotameric side chains in PDB models can clearly be fit to density at a single rotameric conformation and in some cases multiple rotameric conformations; (3) a further 47% of non-rotameric side chains have highly dispersed electron density, indicating potentially interconverting rotameric conformations; (4) the entropy of these side chains is close to that of side chains annotated as having more than one chi(1) rotamer in the crystallographic model; (5) many rotameric side chains with high entropy clearly show multiple conformations that are not annotated in the crystallographic model. These results indicate that modeling of side chains alternating between rotamers in the electron density is important and needs further improvement, both in structure determination and in structure prediction. PMID- 17080463 TI - Cleland's and Grayson's ligaments of the hand: a morphometrical investigation. AB - The cutaneous ligaments of the human digits are delicate functional structures essential for normal skin stability during digital movements. These ligaments extend bilaterally between the phalanx and the finger dermis, either posteriorly (Cleland's ligaments) or anteriorly (Grayson's ligaments) to the digital neurovascular bundles. We have performed a series of detailed anatomical dissections of the human digits so as to investigate morphometrically Cleland's and Grayson's ligaments and their topographic arrangements. Data were statistically compared between fingers, respecting both side (left or right) and sex, in an attempt to clarify some of the morphologic variations of these structures. The cutaneous ligaments of the human digits have been analyzed bilaterally both in 30 fixed cadavers (300 adult human digits) and in 10 nonfixed human cadaveric digits. A computerized morphometrical investigation of the human digits and their Cleland's and Grayson's ligaments has been performed and the resulting quantitative data have been statistically assessed, comparing groups according to finger, phalanx, side (left or right hands), and sex. The ratio between the origin and insertion (O:I) of these ligaments indicate a divergent arrangement of fibers, with values varying from 0.52 to 0.84, depending on the phalanx and finger analyzed. Our morphometrical data provide normal reference values, mainly for Grayson's ligaments, that can be useful in the comparison with the respective measurements obtained in Dupuytren's disease. Morphological bases are also provided, which may be relevant either in computerized tomography or magnetic resonance imaging involving the hand region and in their application in surgical procedures of the human hand. PMID- 17080464 TI - Fatal Parkinsonism-hyperpyrexia syndrome in a Parkinson's disease patient while actively treated with deep brain stimulation. PMID- 17080465 TI - Reduced myocardial 123I-MIBG uptake in a patient with idiopathic rapid eye movement sleep behavior disorder. PMID- 17080466 TI - Bilateral mirror writing movements (mirror dystonia) in a patient with writer's cramp: Functional correlates. PMID- 17080467 TI - Mitral valve hemodynamic effects of percutaneous edge-to-edge repair with the MitraClip device for mitral regurgitation. AB - INTRODUCTION: The Endovascular Valve Edge-to-Edge REpair STudies (EVEREST) are investigating a percutaneous technique for edge-to-edge mitral valve repair with a repositionable clip. The effects on the mitral valve gradient (MVG) and mitral valve area (MVA) are not known. METHODS: Twenty seven patients with moderate to severe or severe mitral regurgitation (MR) were enrolled. Echocardiography was performed preprocedure, at discharge, and at 1, 6, and 12 months. Mean MVG was measured by Doppler and MVA by planimetry and pressure half-time, and evaluated in a central core laboratory. Pre- and postclip deployment, simultaneous left atrial/pulmonary capillary wedge and left ventricular pressures were obtained in eight patients. RESULTS: Three patients did not receive a clip, six patients had their clip(s) explanted by 6 months (none for mitral stenosis), and four were repaired with two clips. Results are notable for a slight increase in mean MVG by Doppler postclip deployment (1.79 +/- 0.89 to 3.31 +/- 2.09 mm Hg, P < 0.01) and an expected decrease in MVA by planimetry (6.49 +/- 1.61 to 4.46 +/- 2.14 cm(2), P < 0.001) and by pressure half time (4.35 +/- 0.98 to 3.01 +/- 1.42 cm(2), P < 0.05). There were no significant changes in hemodynamic parameters postclip deployment by direct pressure measurements. There was no change in MVA by planimetry from discharge to 12 months (3.90 +/- 1.90 to 3.79 +/- 1.54 cm(2), P = 0.78). CONCLUSIONS: Echocardiographic and hemodynamic measurements after percutaneous mitral valve repair with the MitraClip show an expected decrease in mitral valve area with no evidence of clinically significant mitral stenosis either immediately after clip deployment or after 12 months of follow-up. PMID- 17080468 TI - Trends in percutaneous versus surgical revascularization of unprotected left main coronary stenosis in the drug-eluting stent era: a report from the American College of Cardiology-National Cardiovascular Data Registry (ACC-NCDR). AB - OBJECTIVES: We sought to determine how practice patterns for unprotected left main stenosis have changed with the advent of drug-eluting stents (DES). BACKGROUND: Percutaneous coronary intervention (PCI) of unprotected left main coronary stenosis has been controversial. METHODS: We analyzed data submitted to the American College of Cardiology-National Cardiovascular Data Registry (ACC NCDR) between January 1, 2002 and December 31, 2004 from 1,276,582 cardiac catheterizations at 417 institutions. Of these, 53,548 (4.2%) had left main stenosis >50% and no prior CABG. After excluding the unrevascularized, the patient sample (N = 32,562) was analyzed for PCI vs. CABG. Data was stratified by year/quarter, bare metal stent vs. DES, elective vs. urgent/emergent situations, LVEF < or > or =40%, and %left main and RCA stenosis. RESULTS: Of unprotected left main revascularizations from 2002 to 2004, PCI increased from 17.0% to 21.9%, while CABG decreased from 83.0% to 78.1% (P < 0.0001). In 2002, bare metal stents were used for all PCIs; in 2004, bare metal stent use was only 25.5%, while DES use was 74.5% (P < 0.0001). Of elective procedures, PCI rose from 19.1% to 27.5% while CABG fell from 80.9% to 72.5% (P < 0.0001). Similar trends, all significant, were seen in every clinical situation. CONCLUSIONS: In the era of DES, the rate of PCI for unprotected left main stenosis has risen, while CABG has declined. These findings are seen across varying clinical situations, including elective procedures. DES have rapidly and largely replaced bare metal stents for PCI of unprotected left mains. However, PCI is still chosen less frequently than CABG for unprotected left main revascularization. PMID- 17080469 TI - Small, dense low-density-lipoproteins and the metabolic syndrome. AB - Small, dense low-density-lipoproteins (LDL) are associated with increased risk for cardiovascular diseases and diabetes mellitus and a reduction in LDL size has been reported in patients with coronary and non-coronary forms of atherosclerosis. LDL size has been accepted as an important predictor of cardiovascular events and progression of coronary artery disease as well as an emerging cardiovascular risk factor by the National Cholesterol Education Program Adult Treatment Panel III. Small, dense LDL, with elevated triglyceride levels and low HDL-cholesterol concentrations, constitute the 'atherogenic lipoprotein phenotype (ALP)', a form of atherogenic dyslipidemia that is a feature of type 2 diabetes and the metabolic syndrome. LDL size and subclasses show specific alterations in patients with the metabolic syndrome that probably significantly increase their cardiovascular risk; however, so far it has not been recommended to incorporate LDL size measurements in treatment plans, when hypolipidemic therapies are installed. Patients with type 2 diabetes are at high cardiovascular risk and it is still on debate if the treatment goals may be identical or whether there are distinct groups with different cardiovascular risks and hence with different treatment goals. Measurements beyond traditional lipids, such as measurements on the presence of small, dense LDL in patients with the metabolic syndrome, may help to identify cardiovascular risk subgroups. In addition, it might be possible in the future to individualize hypolipidemic treatments if more than the traditional lipids are taken into account. LDL size measurement may potentially help to assess cardiovascular risk within the metabolic syndrome and adapt the treatment goals thereafter. PMID- 17080470 TI - Nanoparticle-based energy transfer for rapid and simple detection of protein glycosylation. PMID- 17080471 TI - Rapid in vivo fingerprinting of nonvolatile compounds in breath by extractive electrospray ionization quadrupole time-of-flight mass spectrometry. PMID- 17080472 TI - Regio- and chemoselective enzymatic N-oxygenation in vivo, in vitro, and in flow. PMID- 17080473 TI - Cation-selective exhaustive injection and sweeping MEKC for direct analysis of methamphetamine and its metabolites in urine. AB - Direct analysis of methamphetamine, amphetamine, and p-hydroxymethamphetamine in urine was achieved by cation-selective exhaustive injection and sweeping micellar EKC. A bare fused-silica capillary (40 cm, 50 microm id) was filled with phosphate buffer (80 mM, pH 3, containing 20% ACN). Then a high-conductivity buffer (100 mM phosphate, pH 3; 6.9 kPa for 2.5 min) was injected. Samples were loaded using electrokinetic injection (10 kV, 600 s) which created long zones of cationic analytes. To enhance sensitivity by sweeping, the stacking step was performed using a phosphate buffer (50 mM, pH 3, containing 20% ACN and 100 mM SDS) at -20 kV before separation by MEKC. This method was capable of detecting the analytes at ppb levels. The calibration plots were linear (r(2) >or= 0.9948) over a range of 100-5000 ng/mL for methamphetamine, and 100-2000 ng/mL for amphetamine and p-hydroxymethamphetamine. The LODs (S/N = 3) were 20 ng/mL for methamphetamine, and 15 ng/mL for amphetamine and p-hydroxymethamphetamine. The method was applied to analysis of 14 urine samples of addicts and is suitable for screening suspected samples for forensic purposes. The results showed good agreement with fluorescence polarization immunoassay and GC-MS. PMID- 17080474 TI - Trace analysis of aminoglycoside antibiotics in bovine milk by MEKC with LIF detection. AB - This work describes a straightforward and sensitive method for the multi-residue analysis of aminoglycoside antibiotics (kanamycin B, amikacin, neomycin B and paromomycin I) in bovine milk samples. The method involves the pre-capillary derivatization of antibiotics with sulfoindocyanine succinimidyl ester (Cy5) and their separation and determination by MEKC with LIF detection. The optimum procedure includes a derivatization step of the antibiotics at 25 degrees C for 30 min and direct injection for MEKC analysis, which is performed in about 20 min by using borate buffer (35 mM; pH 9.2) with 55 mM SDS as an anionic surfactant and 20% ACN as the organic modifier. Under these conditions, dynamic ranges of 10 500 microg/L and RSDs (within-day precision) from 3.8 to 5.3% were obtained. These results indicate that the proposed MEKC-LIF method is useful as a selective and sensitive tool for the determination of these antibiotics and surpasses other reported electrophoretic alternatives. Finally, the method was successfully applied to bovine milk samples after a simple solid-phase extraction clean-up and preconcentration procedure. The aminoglycosides were readily detected at 0.5-1.5 microg/kg levels with average recoveries ranging from 89.4 to 93.3%. PMID- 17080475 TI - Penicillin G acylase as chiral selector in CE using a pullulan-coated capillary. AB - In the present study, penicillin G acylase (PGA), an enzyme belonging to the family of hydrolases, has been investigated as chiral selector in CE using the partial filling technique. Owing to the strong disposition of PGA to be adsorbed by the inner capillary wall, permanently coated capillaries were used to diminish both the protein-wall interactions and the EOF. In particular, the silica surface of the capillary was chemically coated by an antiadhesive and an hydrophilic layer of pullulan, a high-molecular-mass homopolysaccharide. The coating procedure consisted in the silanization with glycidoxypropyltrimethoxysilane and the subsequent coupling of the hydroxyl groups of pullulan onto the silanized capillary. Using this approach, a significant EOF suppression was obtained within a wide pH range (pH 3.0-9.0); this result was very important in order to find the suitable conditions for the application of partial filling technique. The optimization of partial filling was carried out by considering the effects of different experimental conditions (buffer pH, PGA concentration, and loading duration), on the migration time and enantioresolution of rac-ketoprofen. Under the selected conditions as: 100 mM sodium phosphate buffer (pH 5.5) containing 240 microM of PGA (partial filling of 120 s at a pressure of 50 mbar), a series of acidic compounds resulted to be enantioresolved in about 10 min. The long-term stability of the proposed coating was evaluated; more than 100 injections were performed without significant loss of reproducibility. PMID- 17080476 TI - Miniaturized liquid core waveguide-based fluorimetric detection cell for capillary separation methods: application in CE of amino acids. AB - A miniaturized post-column fluorimetric detection cell for capillary separation methods based on optical fibers and liquid core waveguides (LCWs) is described. The main part of the detection cell is a fused-silica capillary coated with Teflon AF serving as an LCW. The optical fibers are used both for coupling the excitation source with the detection domain in the LCW and for the axial fluorescence collection from the LCW end. The latter fiber is connected with a compact CCD spectrometer that serves for the rejection of the scattered excitation light and for the fluorescence signal detection. The proposed design offers a compact fluorescence detector for various microcolumn separation techniques without optical elements such as filters or objectives. Moreover, its construction and optical adjustment are very simple and the whole system is highly miniaturized. The function of the detection cell is demonstrated by CE of amino acids labelled by fluorescein-based tags. Separations of different standard amino acid mixtures and plasma samples are presented. The comparison of plasma amino acid levels of individuals being in good health with those of patients with inherited metabolic disorders is also shown. PMID- 17080477 TI - Eigenmobilities in background electrolytes for CZE. V. Intensity (amplitudes) of system peaks. AB - We present a mathematical model of CZE based on the concept of eigenmobilities - the eigenvalues of matrix M tied to the linearized governing equations of electromigration, and the spectral decomposition of matrix M into matrices of amplitudes P(j). Any peak in an electropherogram, regardless of whether it is an analyte peak or a system peak (system zone), is matched with its matrix P(j). This enables calculation of the peak parameters, such as the transfer ratio and the molar conductivity detection response (which give the indirect detection signal and the conductivity detection signal, respectively), when the initial disturbance caused by the injection of the sample is known. We also introduce new quantities, such as the generalized transfer ratio and the conductivity response of system zones, and show how the amplitude (intensity, area) of the analyte peaks and the system peaks can be calculated. We offer a free software, PeakMaster (http://www.natur.cuni.cz/gas), which yields this information in a user-friendly way. PMID- 17080478 TI - Cooperative coexistence: effective interplay of two Bronsted acids in the asymmetric synthesis of isoquinuclidines. PMID- 17080480 TI - Diffusion of gold into InAs nanocrystals. PMID- 17080479 TI - Proteomic analysis of reporter genes for molecular imaging of transplanted embryonic stem cells. AB - Study of stem cells may reveal promising treatment for diseases. The fate and function of transplanted stem cells remain poorly defined. Recent studies demonstrate that reporter genes can monitor real-time survival of transplanted stem cells in living subjects. We examined the effects of a novel and versatile triple fusion (TF) reporter gene construction on embryonic stem (ES) cell function by proteomic analysis. Murine ES cells were stably transduced with a self-inactivating lentiviral vector containing fluorescence (firefly luciferase; Fluc), bioluminescence (monomeric red fluorescence protein; mRFP), and positron emission tomography (herpes simplex virus type 1 truncated thymidine kinase; tTK) reporter genes. Fluorescence-activated cell sorting (FACS) analysis isolated stably transduced populations. TF reporter gene effects on cellular function were evaluated by quantitative proteomic profiling of control ES cells versus ES cells stably expressing the TF construct (ES-TF). Overall, no significant changes in protein quantity were observed. TF reporter gene expression had no effect on ES cell viability, proliferation, and differentiation capability. Molecular imaging studies tracked ES-TF cell survival and proliferation in living animals. In summary, this is the first proteomic study, demonstrating the unique potential of reporter gene imaging for tracking ES cell transplantation non-invasively, repetitively, and quantitatively. PMID- 17080481 TI - Proteome analysis of maize pollen for allergy-relevant components. AB - Over the last few decades, the cultivation of maize (Zea mays) has strongly increased in Central Europe. We therefore decided to study the allergen composition and the allergenic potency of its pollen in comparison with pollen from timothy grass (Phleum pratense), a typical representative of the native grasses. We found that 65% of the sera reactive to timothy pollen also bound to maize pollen proteins. By using 2-DE immunoblotting, followed by incubation with mAbs directed against known allergens or protein sequencing, those IgE-reactive components were further classified. Although novel, maize-specific pollen allergens could not be found, the presence of crossreacting allergens belonging to groups 1 and 13 (Zea m 1 and 13), both having high IgE prevalence, as well as the presence of the less important group 3 and 12 allergens was found. The structural variability of Zea m 1 and Zea m 13 was determined by sequencing clones isolated from a maize pollen cDNA library. This revealed sequence identities of 72 and 70%, respectively, to the corresponding Phl p 1 and Phl p 13 allergens of timothy grass pollen. IgE-crossreactivity was further studied using immunoblot inhibition tests. Here, timothy pollen extract completely blocked IgE binding to maize, whereas maize pollen extract blocked IgE reactivity to only some timothy pollen allergens. PMID- 17080482 TI - Analysis of the synaptic vesicle proteome using three gel-based protein separation techniques. AB - Synaptic vesicles are key organelles in neurotransmission. Their functions are governed by a unique set of integral and peripherally associated proteins. To obtain a complete protein inventory, we immunoisolated synaptic vesicles from rat brain to high purity and performed a gel-based analysis of the synaptic vesicle proteome. Since the high hydrophobicity of integral membrane proteins hampers their resolution by gel electrophoretic techniques, we applied in parallel three different gel electrophoretic methods for protein separation prior to MS. Synaptic vesicle proteins were subjected to either 1-D SDS-PAGE along with nano LC ESI-MS/MS or to the 2-D gel electrophoretic techniques benzyldimethyl-n hexadecylammonium chloride (BAC)/SDS-PAGE, and double SDS (dSDS)-PAGE in combination with MALDI-TOF-MS. We demonstrate that the combination of all three methods provides a comprehensive survey of the proteinaceous inventory of the synaptic vesicle membrane compartment. The identified synaptic vesicle proteins include transporters, soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs), synapsins, rab and rab-interacting proteins, additional guanine nucleotide triphosphate (GTP) binding proteins, cytoskeletal proteins, and proteins modulating synaptic vesicle exo- and endocytosis. In addition, we identified novel proteins of unknown function. Our results demonstrate that the parallel application of three different gel-based approaches in combination with mass spectrometry permits a comprehensive analysis of the synaptic vesicle proteome that is considerably more complex than previously anticipated. PMID- 17080483 TI - Glycoproteome profiling of transforming growth factor-beta (TGFbeta) signaling: nonglycosylated cell death-inducing DFF-like effector A inhibits TGFbeta1 dependent apoptosis. AB - Transforming growth factor-beta (TGFbeta) is a potent regulator of cell growth, differentiation, and apoptosis. TGFbeta binds to specific serine/threonine kinase receptors, which leads to activation of Smad-dependent and Smad-independent signaling pathways. O-Glycosylation is a dynamic PTM which has been observed in many regulatory proteins, but has not been studied in the context of TGFbeta signaling. To explore the effect of TGFbeta1 on protein O-glycosylation in human breast epithelial cells, we performed analyses of proteins which were affinity purified with Helix pomatia agglutinin (HPA). HPA lectin allowed enrichment of proteins containing GalNAc and GlcNAc linked to serine and threonine residues. Using 2-DE and MALDI-TOF-MS, we identified 21 HPA-precipitated proteins, which were affected by treatment of cells with TGFbeta1. Among these proteins, regulators of cell survival, apoptosis, trafficking, and RNA processing were identified. We found that TGFbeta1 inhibited the appearance of cell death inducing DFF-like effector A (CIDE-A) in 2-D gels with HPA-precipitated proteins. CIDE-A is a cell death activator which promotes DNA fragmentation. We observed that TGFbeta1 did not affect expression of CIDE-A, but inhibited its glycosylation. We found that deglycosylation of CIDE-A correlated with enhanced nuclear export of the protein, and that high level of nonglycosylated CIDE-A inhibited TGFbeta1-dependent cell death. Thus, inhibition of the glycosylation of CIDE-A may be a mechanism to protect cells from apoptosis. PMID- 17080484 TI - Detection in human saliva of different statherin and P-B fragments and derivatives. AB - Statherin is a multifunctional polypeptide specific of human saliva involved in oral calcium homeostasis, phosphate buffering and formation of protein networks. Salivary P-B peptide is usually included into the basic proline-rich protein family but it shows some similarities with statherin and its specific biological role is still undefined. In this study, various fragments and derivatives of statherin and P-B peptide were consistently detected by RP-HPLC ESI-IT MS in 23 samples of human saliva. They were: statherin mono- and non-phosphorylated, statherin Des-Phe(43) (statherin SV1), statherin Des-Thr(42),Phe(43), statherin Des-Asp(1), statherin Des(6-15) (statherin SV2), statherin Des(1-9), statherin Des(1-10), statherin Des(1-13) and P-B Des(1-5). Statherin SV3 (statherin Des(6 15), Phe(43)) was detected only in one sample. Identity of the fragments was confirmed either by MS/MS experiments or by enzymatic digestion or by Edman sequencing. Detection of the fragments suggests that statherin and P-B peptide are submitted to post-translational proteolytic cleavages that are common to other classes of salivary proteins. PMID- 17080485 TI - Simultaneous contactless conductivity detection and UV detection for the study of separation of tamsulosin enantiomers in discontinuous electrolyte systems by CE. AB - This work shows the potential of using discontinuous electrolyte systems for the separation of tamsulosin enantiomers by CE. Sulfated beta-cyclodextrin was used as a chiral selector. In acidic electrolytes, sulfated beta-cyclodextrin migrates as an anion and the analyte (tamsulosin) migrates as a cation. Due to this, four experimental arrangements were proposed. These arrangements differ in composition of electrolytes in the inlet compartment, in the capillary and in the outlet compartment. The separation of tamsulosin enantiomers in acetate buffers with sodium and Tris counterions was studied. Simultaneous contactless conductivity detection and UV detection were used for the study of the separation mechanism in these systems. Mobilities of sulfated beta-cyclodextrin were used for the calculation of the time when the analyte migrates through the BGE zone with the selector. The simulation program Simul 4.0 was used for the calculations of the concentration profiles of the electrolyte components dependent on the time of the separation. The mechanism of enantioseparation in these arrangements was suggested. PMID- 17080486 TI - CEC with monolithic poly(styrene-divinylbenzene-vinylsulfonic acid) as the stationary phase. AB - In this study, poly(styrene-divinylbenzene-vinylsulfonic acid) monolithic columns, prepared by singlestep in situ polymerization, were first used as separation columns for CEC in which vinylsulfonic acid was employed as the charge bearing monomer. The effects of the polymerization conditions (compositions of monomer and porogenic solvent, and polymerization time) on the chromatographic property of the polystyrene-based monolithic column were examined. Furthermore, an increase in water content of the porogenic solvent markedly altered the retention behaviors of these analytes, where the monolithic column was able to be produced in a higher ratio of vinylsulfonic acid. Finally, the study demonstrated that acidic analytes could be successfully separated by the polystyrene-based monolithic column under an acidic mobile phase. PMID- 17080487 TI - Analysis of biologically active amines by CE. AB - This paper provides an overview on the current status of the analysis of biogenic amines by CE. The basic CE separation and detection strategies for the analysis of biogenic amines are briefly described. CZE and MEKC that provide highly efficient and reproducible analysis of biogenic amines are particularly surveyed. With respect to the detection of biogenic amines, we focus on LIF, UV-visible absorption, electrochemiluminescence, and MS. Derivatization strategies, indirect methods, and on-line concentration techniques such as field-amplified sample stacking, sweeping, and use of polymer solution are described. To show the practicality of CE, we highlight currently developed techniques for the determinations of biogenic amines in biological samples, including foods, beverages, cerebrospinal fluids, urine, and single cells. PMID- 17080488 TI - Determination of pK(a) values of diastereomers of phosphinic pseudopeptides by CZE. AB - A CE method was used for the determination of acidity constants (pK(a)) of a series of ten phosphinic pseudopeptides, which varied in number and type of ionogenic groups. Effective electrophoretic mobilities were measured in the 1.8 12.0 pH range in the BGEs of constant ionic strength of 25 mM. Effective electrophoretic mobilities, corrected to standard temperature of 25 degrees C, were subjected to non-linear regression analysis and the obtained apparent pK(a) values were recalculated to thermodynamic pK(a)'s by extrapolation to zero ionic strength according to the extended Debye-Huckel model. The pK(a) values of the phosphinic acid group fell typically in the 1.5-2.25 interval, C-terminal carboxylic groups in the 2.94-3.50 interval, carboxylic groups of the lateral chain of glutamate and aspartate in the 4.68-4.97 interval, imidazolyl moiety of histidine in the 6.55-8.32 interval, N-terminal amino groups in the 7.65-8.28 interval and epsilon-amino group of the lateral chain of lysine in the 10.46 10.61 interval. Further, separation of diastereomers of the phosphinic pseudopeptides was investigated in achiral BGEs. Evaluation of the resolution of the diastereomers as a function of pH of the BGE revealed that most suitable pH region for separation of the diastereomers is around the pK(a) values of the central phosphinic acid group of the pseudopeptides. Successful separation of some diastereomers was, however, achieved in the neutral and alkaline BGEs as well. PMID- 17080489 TI - Chiral resolution of monosaccharides as 1-phenyl-3-methyl-5-pyrazolone derivatives by ligand-exchange CE using borate anion as a central ion of the chiral selector. AB - Six reducing monosaccharides (mannose, galactose, fucose, glucose, xylose, and arabinose) were derivatized with 1-phenyl-3-methyl-5-pyrazolone (PMP) and chiral resolution of these racemic PMP-monosaccharides was studied by ligand-exchange CE using borate anion as a central ion of the chiral selector and (S)-3-amino-1,2 propanediol (SAP) as a chiral selector ligand. PMP-mannose, PMP-galactose and PMP fucose were successfully enantioseparated. Lowering the capillary temperature increased the resolution of PMP-mannose system, but decreased that of PMP galactose and PMP-fucose systems. Whereas the maximum resolution was obtained at pH 8.9 in the PMP-mannose system, resolution increased gradually with pH in the PMP-galactose and PMP-fucose systems. Expecting the formation of the ternary borate complexes with SAP and PMP-monosaccharide in the CE experiments, the optimized structures of the borate diastereomers were obtained by semiempirical molecular orbital calculations to discuss the structural difference of the diastereomers in connection with the enantioseparation behaviors. PMID- 17080490 TI - Screening of drug metabolism by CE. AB - The use of CE for rapid assessment of metabolic stability of drugs with cytochrome P450 (CYP) enzymes, based on relative rates of reduced nicotinamide adenine dinucleotide phosphate (NADPH) consumption and nicotinamide adenine dinucleotide phosphate (NADP) production, was investigated. The separation conditions were as follows: capillary, 80.5 cm (75 microm id, 72 cm effective length for UV detection, 58 cm effective length for fluorescence detection); 25 mM sodium phosphate buffer (pH 8.8); 28 kV (80 microA) applied voltage; UV, 260 nm; fluorescence detection, excitation wavelength, 310 nm, emission wavelength, 418 nm; capillary temperature, 25 degrees C. For UV detection, the incubation conditions were as follows: CYP3A4: 20 pmol/mL; NADPH: 1 mM; EDTA: 1 mM; concentration of the substrate: 5-10 times its reported literature K(m) value; temperature: 37 degrees C; incubation time: 15 min. For fluorescence detection, the concentrations were reduced to CYP3A4: 4 pmol/mL, NADPH: 20 microM, EDTA: 20 microM and substrate: 10 microM. Blank incubations were performed in the absence of substrate. Compared with the blank, significant differences were found for the consumption of NADPH and the production of NADP. The development of this assay system allows rapid assessment of metabolic stability relative to standard compounds, as well as potential identification of the major CYP involved in the metabolism. It would reduce the backlog of compounds that require LC/MS analysis, and thereby expedite the process of metabolic stability screening. PMID- 17080491 TI - Increasing the sample size during clinical trials with t-distributed test statistics without inflating the type I error rate. AB - In clinical trials with t-distributed test statistics the required sample size depends on the unknown variance. Taking estimates from previous studies often leads to a misspecification of the true value of the variance. Hence, re estimation of the variance based on the collected data and re-calculation of the required sample size is attractive. We present a flexible method for extensions of fixed sample or group-sequential trials with t-distributed test statistics. The method can be applied at any time during the course of the trial and does not require the necessity to pre-specify a sample size re-calculation rule. All available information can be used to determine the new sample size. The advantage of our method when compared with other adaptive methods is maintenance of the efficient t-test design when no extensions are actually made. We show that the type I error rate is preserved. PMID- 17080492 TI - Complete assignment of 1H and 13C NMR spectra of some alpha-arylthio and alpha arylsulfonyl substituted N-methoxy-N-methyl propionamides. AB - The complete assignments of the 1H and 13C NMR spectra of the some alpha-arylthio and alpha-arylsulfonyl substituted N-methoxy-N-methyl propionamides, bearing methoxy, methyl, chloro, and nitro as substituents at the phenyl ring are reported. PMID- 17080493 TI - An anomalous course of the reduction of 2-(3-oxo-3,4-dihydroquinoxalin-2 yl)benzene diazonium salt: a reinvestigation. AB - The 1H,13C and 15N NMR spectra of the reduction product of 2-(3-oxo-3,4 dihydroquinoxalin-2-yl)benzene diazonium salt with sodium sulfite were measured and analysed. It is shown that the reaction product corresponds to 1-(indazol-3 yl)-1,2-dihydro-benzimidazol-2-on and not 6H-quinoxalino[1,2-c] [1,2,3]benzotriazin-12(13H)-one as published previously. The correctness of the structure was confirmed by an independent synthesis. The observed 15N chemical shifts were compared with the predicted ones using the ACD/NNMR 9.01 program. PMID- 17080495 TI - Characterization of in vitro metabolites of toad venom using high-performance liquid chromatography and liquid chromatography-mass spectrometry. AB - The characterization of the in vitro metabolites of toad venom, which has been widely used as a traditional Chinese drug, Ch'an Su, has been completed. Toad venom contains bufotoxins (such as marinobufotoxin; marinobufagin 3 suberoylarginine ester) and bufogenins (such as marinobufagin and bufalin) as the main cardiac steroids. An in vitro experiment using the rat or human liver cytosolic fraction disclosed that marinobufotoxin produced marinobufagin, but not its 3-hemisuberate. Marinobufagin was subjected to the enzyme reaction using the rat or human liver microsomal fraction together with NADPH and NAD, which produced 3-dehydromarinobufagin and 3-epimarinobufagin. Marinobufagin produced its 3-sulfate upon treatment with the rat or human liver cytosolic fraction and 3'-phosphoadenosine 5'-phosphosulfate. Bufalin was also subjected to the above enzyme reactions and showed almost the same results except for the result that the hydroxylation occurred at the 5beta-position. On the other hand, small amounts of marinobufagin 3-glucuronide were obtained only by treatment with the human liver microsomal fraction and uridine 5'-diphosphoglucuronic acid. The structures of these metabolites were confirmed using authentic samples regarding their high-performance liquid chromatographic behavior and/or liquid chromatography-mass spectrometry analysis. PMID- 17080494 TI - Extending the validity of the Memorial Anxiety Scale for Prostate Cancer (MAX-PC) at the time of prostate biopsy in a racially-mixed population. AB - The Memorial Anxiety Scale for Prostate Cancer (MAX-PC) has been validated for assessing men with prostate cancer for cancer-specific anxiety. It was originally validated in a predominantly white population. The MAX-PC Prostate Cancer Anxiety Subscale (MAX-PC-PCAS) may be relevant for measuring cancer-specific anxiety in undiagnosed men at risk for prostate cancer. We assess the validity of the MAX-PC PCAS at the time of prostate biopsy (n = 178). Questions assessed socio demographic information, health status, patient-estimated risk of cancer, the Hospital Anxiety and Depression Scale--Anxiety Subscale (HADS-A), and the MAX-PC PCAS. The patients' most recent PSA was recorded. Cronbach's alpha, inter-item correlations, and Pearson correlations with both the HADS-A and clinical variables were compared with the original validation sample. Our sample was younger (63.1 vs 71.1 years), had a larger fraction of African-Americans (43 vs 10%), and had higher PSAs. Cronbach's alpha was equivalent (0.91 vs 0.90), median inter-item correlation was equivalent (0.63 vs 0.61), and Pearson correlation with HADS-A was higher (0.71 vs 0.57). Anxiety levels were not correlated with PSA levels, and there were minor differences in the validation findings by race. The validity of the MAX-PC-PCAS extends to men without cancer undergoing biopsy and to African-Americans. PMID- 17080496 TI - Analysis of alkannin derivatives from Alkanna species by high-performance liquid chromatography/photodiode array/mass spectrometry. AB - Alkannin, shikonin (A/S) and their derivatives are enantiomeric hydroxynaphthoquinone red pigments found in the roots of almost 150 species of the Boraginaceae family. A/S have been shown to exhibit strong wound healing, antimicrobial, anti-inflammatory and antioxidant activities and recent extensive research has well established their antitumor properties. A/S and their derivatives comprise the active ingredients of several pharmaceutical and cosmetic preparations. Although A/S have been efficiently synthesized and have been produced by cell tissue cultures in high yield, most of the pharmaceutical preparations worldwide contain A/S extracted from the roots of Boraginaceous species, found in nature. In the present study, a high-performance liquid chromatography/photodiode array/mass spectrometry (HPLC/PDA/MS) method was established to identify monomeric hydroxynaphthoquinones of the alkannin series and other metabolites from Boraginaceous root extracts. This method can be applied for the identification of alkannin derivatives and other metabolites from Boraginaceous cell cultures, and also to determine active ingredients in pharmaceutical preparations containing A/S derivatives. A phytochemical investigation of the Alkanna genus grown in Greece was also performed. Fifty three root samples belonging to 10 species of the genus Alkanna (A. calliensis, A. corcyrensis, A. graeca, A. methanaea, A. orientalis, A. pindicola, A. primuliflora, A. sieberi, A. stribrnyi and A. tinctoria) were collected from several regions of the Greek flora and analyzed for their constituent hydroxynaphthoquinones and other metabolites. In most of the above Alkanna samples tested, the main hydroxynaphthoquinones were determined to be beta,beta dimethylacrylalkannin, isovalerylalkannin + alpha-methyl-n-butylalkannin and acetylalkannin. The hydroxynaphthoquinone constituents and their proportions were found to vary among Alkanna species. Unknown metabolites (not monomeric hydroxynaphthoquinones) were detected by HPLC-PDA-MS, while in several Alkanna species hydroxynaphthoquinones were detected for the first time. PMID- 17080497 TI - Simple determination of huperzine A in human plasma by liquid chromatographic tandem mass spectrometric method. AB - Huperzine A is a potent, reversible acetylcholinesterase inhibitor. In the present work, a rapid and sensitive LC-MS-MS method for the determination of huperzine A in human plasma using codeine phosphate as internal standard has been developed and validated. The analyte and internal standard were extracted from plasma using ethyl acetate, chromatographed on a C(18) column (5 microm, 150 x 4.6 mm i.d.) with a mobile phase consisting of 1% formic acid-methanol (40:60, v/v), and detected using a tandem mass spectrometer with a TurboIonSpray ionization interface. The run time was only 2 min. Good linearity was achieved in the range 0.126 -25.2 ng/mL and the limit of detection in plasma was 0.064 ng/mL. The average recovery for huperzine A was 83.4% from plasma. The analytical sensitivity and accuracy of this assay is adequate for characterization of huperzine A in human plasma. PMID- 17080498 TI - Simple determination of pirfenidone in rat plasma via high-performance liquid chromatography. AB - A simple, rapid and reliable high-performance liquid chromatographic method was developed and validated for the determination of pirfenidone and its major metabolites in rat plasma. Plasma proteins were precipitated with perchloric acid (10%, v/v) and the supernatant after centrifugation was determined using high performance liquid chromatography. The analysis was carried out on a Lichrospher C(18) column (250 x 4.6 mm i.d., 5 microm). The mobile phase consisted of acetonitrile-water containing 0.2% acetic acid (23:77, v/v) at a flow-rate of 1 mL/min. The eluant was detected at 310 nm. The calibration curves were linear over a concentration range from 0.15 to 76.67 microg/mL. The accuracy (relative error) of the assay ranged from -2.6 to 7.9% and the precision (coefficient of variation) was less than 4.5%. The established method has been successfully applied to a pharmacokinetic study of pirfenidone following a single oral dose to rats. PMID- 17080499 TI - Comparison between micellar liquid chromatography and capillary zone electrophoresis for the determination of hydrophobic basic drugs in pharmaceutical preparations. AB - The determination of highly hydrophobic basic compounds by means of conventional reversed-phase liquid chromatographic methods has several drawbacks. Owing to the characteristics of micellar liquid chromatography (MLC) and capillary electrophoresis (CE), these techniques could be advantageous alternatives to reversed-phase chromatographic methods for the determination of these kinds of compounds. The objective of this study was to develop and compare MLC and CE methods for the determination of antipsychotic basic drugs (amitryptiline, haloperidol, perphenazine and thioridazine) in pharmaceutical preparations. The chromatographic determination of the analytes was performed on a Kromasil C(18) analytical column; the mobile phase was 0.04 m cetyltrimethylammonium bromide (CTAB), at pH 3, containing 5% 1-butanol, at a flow rate of 1 mL/min. The CE separation was performed in a fused-silica capillary with a 50 mm tris (hydroxymethyl)-aminomethane buffer, pH 7, at an applied voltage of 20 kV, using barbital as internal stardard. The proposed methods are suitable for a reliable quantitation of these compounds in the commercial tablets and drops in terms of accuracy and precision and require a very simple pre-treatment of the samples. By comparing the performance characteristics and experimental details of the MLC and CE methods we conclude that CE seems to be slightly better than MLC in the determination of highly hydrophobic compounds in pharmaceuticals in terms of resolution and economy, taking into account that the limits of detection are not a handicap in pharmaceutical samples. PMID- 17080500 TI - Unexplained acidosis of malnutrition: a study by ion-exchange chromatography/mass spectrometry. AB - Keto-acidosis is usually associated with uncontrolled diabetes and typically poses few diagnostic problems when presenting as hyperglycaemia, metabolic acidosis and a high anion gap. An emaciated patient suffering from Duchenne Muscular Dystrophy and volume depletion presented with acidosis of unknown origin. Preliminary investigations appeared to rule out lactic acidosis, diabetic keto-acidosis and acidosis due to base loss. We have previously reported a technique utilizing liquid chromatography coupled to mass spectrometry (LC-MS) which can be used to characterize the underlying aetiology of acidosis and applied it to ultrafiltrate derived from a blood sample taken from this patient. The anion profile obtained on the chromatogram showed elevated levels of acetoacetate and hydroxybutyrate but no evidence of lactic acidosis, nor was the profile typical of that seen in 'unexplained' acidosis. We concluded that the patient was suffering from keto-acidosis associated with starvation and dehydration, the biochemical features being obscured by both the patient's chronic malnutrition and minimal muscle mass. A combination of enteral feeding and rehydration led to prompt resolution of the patient's metabolic acidosis. PMID- 17080501 TI - Benztropine pretreatment does not affect responses to acute cocaine administration in human volunteers. AB - Benztropine (Cogentin ) was evaluated for its ability to block cocaine's physiological and subjective effects in humans. In healthy, recreational users of cocaine, placebo, or benztropine (1, 2, and 4 mg orally) was given 2 hr before subjects self-administered 0.9 mg/kg of cocaine intranasally. Measurements were made for 2 hr following cocaine administration, and plasma cocaine and cocaine metabolites were assayed. Cocaine produced typical increases in heart rate and alterations in self-reports measured by visual analog scales (VAS). Benztropine alone did not produce changes on any of these measures. Responses to cocaine with and without benztropine pretreatment were similar: benztropine did not change cocaine's effects. This study of one of the tropane-ring analogs that is approved for human use suggests this compound does not alter cocaine-induced effects, but just as importantly, does not produce any adverse behavioral or physiological effects. The exact therapeutic application of benztropine as a possible adjunct treatment for cocaine abuse in humans require further exploration. PMID- 17080502 TI - Colloidal gold-based immunochromatographic assay for detection of diethylstilbestrol residues. AB - One-step membrane-based competitive colloidal gold-based immunoassays in immunochromatographic formats for the rapid detection of diethylstilbestrol (DES) were developed. Nitro-cellulose membrane strip was separately coated with goat anti-rabbit IgG (control line) and DES hapten-ovalubumin conjugate (test line). Anti-DES polyclonal antibody labeled with colloidal gold particles was first incubated with DES. A positive reaction as a result of the remaining antibody gold conjugate combining with antigen coated on the membrane was obvious by visual detection, with detection limits for immunochromatographic of 0.5 microg/kg for detecting DES standard solution, and the limit of detection was 5 microg/kg for detecting the DES spiked in swine pork and liver. The assay time for test was less than 5 min, suitable for rapid testing on-site. PMID- 17080503 TI - High-throughput liquid-chromatography method with fluorescence detection for reciprocal determination of furosemide or norfloxacin in human plasma. AB - A simple, high-throughput, highly selective and sensitive HPLC-FLD method for isolation and determination of furosemide and/or norfloxacin in human plasma samples following a simple organic solvent deproteinization step with acetonitrile as sample 'clean-up' procedure is reported. One of the two drug substances plays the internal standard role for the determination of the other. Separation of analyte and internal standard was achieved in less than 5.3 min (injection to injection) on a Chromolith Performance RP-18e column, using an aqueous component containing 0.015 mol/L sodium heptane-sulfonate and 0.2% triethylamine brought to pH = 2.5 with H(3)PO(4). The composition of the mobile phase was: acetonitrile-methanol-aqueous component = 70:15:15 (v/v/v) and the flow-rate was set up to 3 mL/min. The chromatographic method applied to the determination of furosemide relies on fluorescent detection parameters of 235 nm for the excitation wavelength, and 402 nm for the emission wavelength. In case of norfloxacin, the excitation wavelength is set up to 268 nm and the emission wavelength is set up to 445 nm. The overall method leads to quantitation limits of about 27 ng/mL for furosemide, and 19.5 ng/mL for norfloxacin, using an injection volume of 250 microL. The method was applied to the bioequivalence study of two furosemide-containing formulations. PMID- 17080504 TI - A simple and sensitive HPLC-fluorescence method for quantification of MDMA and MDA in blood with 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride (DIB-Cl) as a label. AB - A sensitive high-performance liquid chromatographic method with fluorescence detection to determine 3,4-methylenedioxymethamphethamine (MDMA) and 3,4 methylenedioxyamphethamine (MDA) in human and rat whole blood or plasma samples was developed by using 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride (DIB-Cl) as a label. MDMA and MDA in a small amount of blood sample (ca 100 microL) were extracted by liquid-liquid extraction with ethyl acetate, and were derivatized with DIB-Cl under mild conditions (10 min at room temperature). A good separation of DIB-derivatives could be achieved within 45 min using a commercially available ODS column with an isocratic eluent of 10 mM citric acid-20 mM Na(2)HPO(4) aqueous buffer (pH 4.0)-CH(3)CN-CH(3)OH (50:45:5, v/v/v %). The calibration curves prepared with 1-methyl-3-phenylpropylamine (MPPA) as an internal standard showed good linearity (r = 0.999) with 0.36-0.83 ng/mL detection limit at a signal-to-noise ratio of 3. MDMA and MDA in rat whole blood could be monitored for 6 h after a single administration of MDMA (2.2 mg/kg, i.p.). The pharmacokinetic parameters for MDMA and MDA obtained by triplicate measurements were 426 +/- 23 and 39 +/- 6 ng/mL (C(max)), 20 +/- 5 and 100 +/- 10 min (T(max)), respectively. PMID- 17080505 TI - Determination of polymethoxylated flavones in peels of selected Jamaican and Mexican citrus (Citrus spp.) cultivars by high-performance liquid chromatography. AB - The concentrations of the polymethoxylated flavones (PMFs) in peels of selected citrus cultivars grown in Jamaica and Mexico were determined. The PMFs were extracted from sun-dried citrus peels with reagent-grade methanol. Analyses were carried out by reverse-phase HPLC and UV detection. The column used was a C(18) 5 microm (150 x 4.6 mm) Discovery column. Elution was in the gradient mode, using a ternary mobile phase. The results showed that all the citrus cultivars used contained at least three of the six major PMFs quantified. Ortanique peel contained the highest quantity of PMFs (34,393 +/- 272 ppm), followed by tangerine (28,389 +/- 343 ppm) and Mexican sweet orange (sample 1; 21,627 +/- 494 ppm). The major PMFs, i.e. sinensetin, nobiletin, tangeretin, heptamethoxyflavone, tetramethylscutellarein and hexamethyl-o-quercetagetin, present in the peels of 20 citrus cultivars, was quantified. The results were compared with those of Florida citrus peels. A large amount of citrus peels and byproducts are produced in the Caribbean which could provide a cheap and convenient source of PMFs. PMID- 17080506 TI - A rapid liquid chromatography-tandem mass spectrometry method for quantification of a biologically active molecule camboginol in the extract of Garcinia cambogia. AB - A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of camboginol in the extract of fruit rinds of Garcinia cambogia has been developed. Separation was achieved isocratically on an RP C(18) column using a solvent system consisting of a mixture of acetonitrile-water (9:1) and methanol-acetic acid (99.5:0.5) in the ratio of 30:70 as mobile phase at a flow rate of 0.4 mL/min. A multiple reaction monitoring (MRM) method was developed for quantification of camboginol in the fruit rinds extract of G. cambogia using MRM transitions of m/z 601.4 --> m/z 176.7 and m/z 601.4 --> m/z 448.9, respectively. The calibration curve based on peak area against concentration was linear up to 50 ng/mL with a detection limit of 0.5 ng/mL. The method showed satisfactory reproducibility with a coefficient of variation of less than 6%. The method was successfully applied for quantification of camboginol in different Garcinia extracts. PMID- 17080507 TI - A rapid and specific approach for direct measurement of pravastatin concentration in plasma by LC-MS/MS employing solid-phase extraction. AB - A rapid, specific and sensitive LC-MS/MS assay using solid-phase extraction (SPE) for the determination of pravastatin, in human plasma is described. The plasma filtrate obtained after SPE, using a polymer base, a hydrophilic-lipophilic balance (HLB) cartridge, was submitted directly to short-column liquid chromatography-tandem mass spectrometric (LC-MS/MS) assay, with negligible matrix effect on the analysis. For validation of the method, the recovery of the free analytes was compared with that from an optimized extraction method, and the analyte stability was examined under conditions mimicking the sample storage, handling, and analysis procedures. The extraction procedure yielded extremely clean extracts with a recovery of 107.44 and 98.93% for pravastatin and IS, respectively. The intra-assay and inter-assay precisions for the samples at the LLOQ were 3.30 and 7.31% respectively. The calibration curves were linear for the dynamic range 0.5-200 ng/mL with correlation coefficient r > or = 0.9988. The intra- and inter-assay accuracy ranged from 95.87 to 112.40%. The method is simple and reliable with a total run time of 3 min. This novel validated method was applied to the pharmacokinetic (PK) study in human volunteers receiving a single oral dose of 40 mg immediate release (IR) formulation. PMID- 17080508 TI - Extraction of paeonol from Jisheng Shenqi Wan using supercritical fluid extraction. AB - Supercritical carbon dioxide with or without methanol and 95% ethanol modifiers was used to extract paeonol from the pills of Jisheng Shenqi Wan and high performance liquid chromatography (HPLC) was used to analyze the extract. The effect of temperature and pressure on the yield of paeonol was tested. The results show that the recovery of paeonol was improved by adding a polar modifier, such as methanol and 95% ethanol in supercritical fluid. The yield obtained after 120 min extraction with 95% ethanol modified-supercritical carbon dioxide was the highest (1.51, w/w at 60 degrees C and 400 bar), while that obtained with supercritical carbon dioxide only at 40 degrees C was the lowest from 0.67 to 0.83 over a range of 200-600 bar. PMID- 17080509 TI - Determination of dopamine and its relativity of baicalin in rat nuclei after intravenous administration of flavonoids from Scutellariae radix. AB - Baicalin is an active component of Scutellariae radix extracts. It can cross the blood-brain barrier and distribute in cerebral nuclei. However, its mechanism and the effects on the nuclei where it accumulates remain unclear. We used an HPLC electrochemical detection method to determine the dopamine in cerebral nuclei after intravenous administration of flavonoids from Scutellariae radix and analyzed the relativity between baicalin and DA in cerebral nuclei. We found that the dopamine system is another target system of brain function which baicalin acts on. PMID- 17080510 TI - HPLC determination and pharmacokinetic study of tenatoprazole in dog plasma after oral administration of enteric-coated capsule. AB - A simple, sensitive and selective high-performance liquid chromatographic (HPLC) method with UV detection (306 nm) was developed and validated for determination of tenatoprazole, a novel proton-pump inhibitor, in dog plasma. Tenatoprazole and internal standard (pantoprazole) were extracted into diethyl ether and separated using an isocratic mobile phase of 10 mm phosphate buffer (pH4.7)-acetonitrile (70:30, v/v) on a Diamonsil C(18) column (150 x 4.6 mm, 5 microm). The retention times for tenatoprazole and internal standard were 7.1 and 12.3 min, respectively. No endogenous interferences were observed. This HPLC method was fully validated. The lower limit of quantitation was 20 ng/mL, with a relative standard deviation of less than 20%. A linear range of 0.02-5.0 microg/mL was established. The interday and intraday precisions were within RSD 13.4-10.1 and 4.6-1.4%, respectively. This method developed can be easily applied to the pharmacokinetic study of tenatoprazole in dog plasma after oral administration of an enteric-coated capsule. The plasma concentration of tenatoprazole from six dogs showed a mean C(max) of 2.63 microg/mL at T(max) of 1.89 h. The bioavailability of tenatoprazole was improved by administration of enteric-coated capsule. PMID- 17080511 TI - Quantitative 2D HSQC NMR determination of polymer structures by selecting suitable internal standard references. AB - A new analytical method based on the 2D HSQC NMR sequence is presented, which can be applied for quantitative structural determination of complicated polymers. The influence of T1 and T2 relaxations, off-resonance effects, coupling constants and homonuclear couplings are discussed. It was found that the T2 values measured on polymeric samples with the conventional HSQC-CPMG sequence could not be used to correct the errors caused by T2 relaxations during the polarization transfer delay. A unique way of selecting the proper internal standard reference signal(s) is therefore proposed to eliminate the major errors caused by T2 relaxations, resonance offsets, coupling constant deviations and homonuclear couplings. Two polymer samples, a cellulose triacetate and an acetylated lignin, have been used to illustrate the principles. The methodology developed in this work is robust to instrument miss-setting and it can find wide-spread applications in areas where a quantitative analysis of structurally complicated polymers is necessary. PMID- 17080512 TI - Resistance and cross-resistance to imidacloprid and thiamethoxam in the Colorado potato beetle Leptinotarsa decemlineata. AB - One of the major challenges in managing the Colorado potato beetle, Leptinotarsa decemlineata (Say) is its remarkable ability to develop resistance to virtually every insecticide that has ever been used against it. Resistance is particularly common throughout northeastern USA as far north as Maine. The first instances of resistance to imidacloprid have already been reported from several locations in New York, Delaware and southern Maine. Rotating insecticides with different modes of action may delay insecticide resistance, but successful implementation of this technique depends on a good understanding of resistance and cross-resistance patterns in populations of target pests. LC(50) values were measured for imidacloprid and thiamethoxam in Colorado potato beetle populations from a variety of locations in the USA and Canada using diet incorporation bioassays. The field performance of imidacloprid, thiamethoxam and clothianidin against imidacloprid-resistant beetles on a commercial potato farm in southern Maine was also evaluated. Correlation between LC(50) values for imidacloprid and thiamethoxam was highly significant, even when populations previously exposed to thiamethoxam were excluded from the analysis. There was no statistically detectable difference in the LC(50) values between populations exposed to both insecticides and to imidacloprid alone. Applications of neonicotinoid insecticides at planting delayed build-up of imidacloprid-resistant beetle populations on field plots by 1-2 weeks but failed to provide adequate crop protection. Consistently with bioassay results, there was also substantial cross resistance among the three tested neonicotinoid insecticides. Results of the present study support the recommendation to avoid rotating imidacloprid with thiamethoxam as a part of a resistance management plan. PMID- 17080513 TI - Geode of the femur: an uncommon manifestation potentially reflecting the pathogenesis of rheumatoid arthritis. AB - Geodes are noted frequently in rheumatoid arthritis (RA), but large geodes of the femur are uncommon. We describe a patient with RA and a large geode in his femur; histological findings were consistent with a rheumatoid nodule and chronically inflamed synovium. We review the literature of large femoral geodes and what this particular manifestation may reflect about the pathogenesis of RA. PMID- 17080515 TI - Bosentan increases myocardial perfusion and function in systemic sclerosis: a magnetic resonance imaging and Tissue-Doppler echography study. AB - OBJECTIVE: To evaluate the short-term effects of bosentan on myocardial perfusion and function assessed by cardiac magnetic resonance imaging (MRI) and Tissue Doppler echography (TDE) respectively in patients with systemic sclerosis (SSc). METHODS: We prospectively evaluated 18 SSc patients without clinical heart failure and with normal pulmonary arterial pressure. MRI perfusion index and systolic and diastolic strain rates (SR) determined by TDE were assessed at baseline for comparison with healthy controls (after a 72-hour vasodilator washout period), and repeated after 4 weeks of bosentan treatment (62.5 mg bid for 2 weeks titrated to 125 mg bid for 2 weeks). RESULTS: Patients with SSc had decreased MRI perfusion index and TDE SR in comparison with healthy controls. Bosentan treatment led to a significant increase in median (range) global MRI perfusion index [from 0.17 (0.09-0.23) at Day 0 to 0.22 (0.13-0.30) after bosentan treatment; p = 0.0004], systolic SR [from 2.1 (1.3-3.1) s-1 to 2.8 (2.1 4.8) s-1; p = 0.0002), and diastolic SR [from 2.6 (1.4-6.7) to 3.6 (2.0-7.6) s-1; p = 0.0003]. CONCLUSION: Short-term treatment with bosentan simultaneously improves myocardial perfusion and function, as evaluated by highly sensitive and quantitative methods, in patients with SSc. Whether additional remodeling effect may be observed after longterm treatment with bosentan remains to be determined. PMID- 17080516 TI - Development and validation of a preference weight multiattribute health outcome measure for rheumatoid arthritis. AB - OBJECTIVE: To develop and validate multiattribute measures for patients with rheumatoid arthritis (RA) to report health states and estimate preference weights. METHODS: Survey materials were mailed to 748 patients. Factor analysis, an item response theory-based model, and an internal consistency test were used to identify attributes and evaluate items. Two multiattribute preference weight functions (MAPWF) were constructed. Construct validity of the new measures was then tested. RESULTS: Four hundred eighty-seven patients returned the survey; 24 items on 6 health attributes were selected to form the new outcomes measure. Two MAPWF were derived with preference weights measured with time tradeoff and visual analog scales as dependent variables. All validity test results were statistically significant. CONCLUSION: Our results reveal that the new measures are reliable and valid in assessing health states and associated preference weights of patients with RA. PMID- 17080517 TI - CD64 on neutrophils is a sensitive and specific marker for detection of infection in patients with rheumatoid arthritis. AB - OBJECTIVE: In inflammatory diseases, differentiation between infection and disease flares is often clinically difficult because of similar signs and symptoms, such as fever and elevation of inflammatory markers. In rheumatoid arthritis (RA), infection is not only one of the major complications but also one of the frequent causes of death. Use of biologic agents such as tumor necrosis factor-a blockers has been reported to increase the incidence of tuberculosis or opportunistic infections. We examined the utility of CD64 (FcgRI) expressed on neutrophils as a marker for detection of infection complicated with RA. METHODS: We measured the expression level of CD64 per neutrophil quantitatively by flow cytometry in 279 samples from 237 patients with RA with various levels of disease activity or types of infection, and in 52 samples from 36 controls including subjects with infection. RESULTS: CD64 expression was significantly higher among RA patients with infection (median 4156 molecules per neutrophil, interquartile range 2583-8587) than in those without infection (884, IQR 670-1262) (p < or = 0.001). The sensitivity of CD64 on neutrophils for the diagnosis of infection (using a cutoff value of 2000 molecules per cell) was 92.7% and specificity was 96.5%. CD64 expression was not affected by the disease activity of RA or the use of corticosteroids, disease modifying antirheumatic drugs, and biologic agents. CD64 was upregulated in infection by bacteria, viruses, fungi, and mycobacteria. CONCLUSION: Our results suggest that quantitative measurement of CD64 expression on neutrophils can be used as a sensitive and specific marker to detect infection complicating RA. PMID- 17080518 TI - An index of patient reported outcomes (PRO-Index) discriminates effectively between active and control treatment in 4 clinical trials of adalimumab in rheumatoid arthritis. AB - OBJECTIVE: To analyze 2 indices composed of the 3 patient reported outcomes (PRO) in the American College of Rheumatology (ACR) Core Data Set--physical function, pain, and global estimate--without joint count or laboratory data, for capacities to distinguish active from control treatments in 4 pivotal clinical trials. METHODS: Data from 4 clinical trials involving adalimumab, in combination with methotrexate or other disease-modifying antirheumatic drugs (DMARD) or as monotherapy, versus control treatment were made available to analyze properties of various indices. A categorical PRO-Index M was defined as "majority" improvement in 2 of the 3 PRO measures at 20%, 50%, and 70% levels; results were evaluated to analyze agreement with ACR20, ACR50, ACR70 responses and an "all Core Data Set measures" index based on 4 of the 7 measures having such levels of improvement. A continuous PRO-Index C was defined as the median or 2nd highest of 3 percentage differences from baseline to endpoint; results were evaluated to analyze agreement with a continuous ACR-N, "all Core Data Set measures" index, and Disease Activity Score 28 (DAS28). RESULTS: All indices distinguished active versus control treatment at similar levels, including PRO-Index M versus ACR20, ACR50, and ACR70 responses, and PRO-Index C versus DAS28. CONCLUSION: PRO indices based only on patient questionnaire data, without joint counts or laboratory tests, may be useful quantitative measures of therapeutic efficacy for use in standard rheumatology clinical care. PMID- 17080519 TI - Metabolic syndrome and subclinical atherosclerosis in rheumatoid arthritis. AB - OBJECTIVE: We assessed whether features of metabolic syndrome (MetSyn) were risk factors for subclinical atherosclerosis independent of previously identified determinants of cardiovascular disease in 74 patients with rheumatoid arthritis (RA). We further evaluated the clinical utility of currently recommended MetSyn definitions in the identification of RA patients with subclinical atherosclerosis. METHODS: We investigated the associations of MetSyn features and MetSyn definitions with ultrasonographically determined common carotid artery intima-media thickness (CCA-IMT) and plaque, with adjustment for age, radiographic scores (cumulative inflammation), polymorphonuclear cell counts (current inflammation), or hypothyroidism. RESULTS: The Quantitative Insulin Sensitivity Check Index (QUICKI) (partial R = -0.24 to -0.26, p = 0.04 to 0.02), log triglycerides (partial R = 0.23 to 0.30, p = 0.05 to 0.01), and systolic blood pressure (partial R = 0.22 to 0.30, p = 0.06 to 0.002) were consistently associated with the log CCA-IMT. Log triglycerides (OR 1.04, 95% CI 1.01-1.08, p = 0.02) and the QUICKI (OR 0.22, 95% CI 0.05-0.91, p = 0.03) were associated with plaque after adjusting for cumulative inflammation. Hypertension (blood pressure > or = 130/85 mm Hg or drug treatment for hypertension) was consistently associated with CCA-IMT (p = 0.05 to 0.0003) and plaque (p = 0.03 to 0.006). The WHO-defined MetSyn was associated with CCA-IMT (p = 0.08 to 0.04) but not with plaque (p > or = 0.1). The National Cholesterol Education Program-defined MetSyn was not associated with CCA-IMT or plaque (p > or = 0.3). CONCLUSION: In this RA cohort, the MetSyn features of hypertension, insulin resistance, and triglycerides were risk factors for subclinical atherosclerosis, independent of previously identified determinants of cardiovascular disease. Individual MetSyn features were more strongly associated with subclinical atherosclerosis than were currently recommended MetSyn definitions. PMID- 17080520 TI - Clinical outcome of extrasynovial steroid injection for trigger finger. AB - A prospective clinical study was performed to investigate the clinical results of extrasynovial (subcutaneous) steroid injection for trigger finger. One hundred and twenty-nine trigger fingers were investigated in 100 adult patients; 76 were women and 24 were men. Their mean age was 60 years (range: 17 to 88 years). We classified trigger fingers into three different grades according to clinical severity at a medical examination. All patients were injected with betamethasone mixed with lidocaine. Surgical release of the A1 pulley was performed at the patients' request if steroid injection therapy was not effective. Pain and snapping were relieved in 98% and 74% of cases, respectively. Recurrence occurred in about half our patients, but the same clinical benefit was obtained after re injection. Surgery was performed for seven fingers. No complications of steroid injections were observed. This study suggests that extrasynovial steroid injection is a valuable conservative treatment for trigger finger and it is not necessary to try and inject into the tendon sheath to get a good result and markedly reduce the risk of causing damage to tendons and other structures. PMID- 17080521 TI - Morbidity of hand and wrist Ganglia. AB - Pain and disability caused by ganglia of the hand and wrist were assessed using a patient-rated wrist evaluation questionnaire in 75 patients. Dorsal wrist ganglia were the most painful and disabling. However, the majority of ganglia cause little pain or disability. Consequently, referral by General Practitioners should be confined to those with pain, disability or failure of conservative management. PMID- 17080522 TI - Limited wrist arthrodesis versus radial osteotomy for advanced Kienbock's disease -for a fragmented lunate. AB - Thirty-eight patients with advanced Kienbock's disease treated by limited wrist arthrodesis (LWA: n = 10) or radial osteotomy (RO: n = 28) for a fragmented lunate were retrospectively examined after an average of 47.9 and 68.1 months, respectively. Compared with pre-operative values, the active flexion-extension range of motion decreased by about 16.0 degrees in LWA and increased approximately 9.7 degrees in RO and the grip strength improved by approximately 7.5 kg in LWA and 8.0 kg in RO. In both groups, radiographs showed no significant progression of carpal collapse. Although LWA caused some decrease in wrist flexion-extension, both procedures are appropriate for surgical treatment of advanced Kienbock's disease. Most patients experienced a reduction in pain and were able to return to work. PMID- 17080523 TI - Modified matched ulnar resection for arthrosis of distal radioulnar joint in rheumatoid arthritis. AB - The pain of distal radioulnar arthrosis in rheumatoid patients is often due to disease largely confined to the radioulnar rather than the ulnocarpal articulation. This is a retrospective study of 14 patients (14 wrists) who underwent selective shaving of the radial articulation of the ulnar head leaving the ulnocarpal articulation intact. The ulnar head is reduced to the circumference of its shaft and a dorsal retinacular flap is interposed between it and the distal radius. Average age of the patients and follow-up were 63.5 years and 31 months, respectively. All patients had rheumatoid arthritis. Pain improved in 14 out of 15 wrists. Overall results were 67% good to excellent and 33% fair based on the modified Mayo's wrist score. This novel procedure for DRUJ arthrosis produces predictable pain relief in low demand rheumatoid patient. PMID- 17080524 TI - Results and complications in dorsal and volar wrist Ganglia arthroscopic resection. AB - The authors present the procedure and results of five years of arthroscopic treatment of wrist radiocarpal and midcarpal ganglia. Thirty cases of dorsal ganglia and seventeen cases of volar ganglia were operated on arthroscopically. The technique was easy to perform in all the radiocarpal ganglia, not easy in midcarpal dorsal ganglia and very difficult in midcarpal volar ganglia. The results were recorded with a mean follow-up of 15 months. Twenty-seven cases of dorsal ganglia and twelve cases of volar ganglia had excellent results with active motion recovery, no complications, absence of scars and no recurrence. Two cases had a recurrence. There were four complications: a case of injury of a radial artery branch, a case of extensive haematoma, and two cases of neuropraxia. In three cases the procedure was converted into open surgery: they had a longer time of healing and a residual scar. The arthroscopic resection has been in our experience effective and safe for the treatment of all radiocarpal ganglia. Good results have been obtained also in the treatment of dorsal midcarpal ganglia. Concerning the uncommon cases of volar midcarpal (STT) ganglia, an open approach seems still indicated. PMID- 17080525 TI - Comparative responsiveness of Japanese versions of the DASH and SF-36 questionnaires and physical measurement to clinical changes after carpal tunnel release. AB - The purpose of the present study was to test the responsiveness of the Japanese Society for Surgery of the Hand version of the Disability of the Arm, Shoulder and Hand questionnaire (DASH-JSSH) by evaluating effect size (ES) and standardised response mean (SRM) in patients undergoing carpal tunnel release. Subjects comprised 25 patients with carpal tunnel syndrome. All subjects completed the DASH-JSSH, medical outcomes 36-item short-form health survey (SF 36) and visual analogue scale (VAS) for pain and underwent objective assessment of grip strength, pinch strength and static two-point discrimination before and three months after surgery. DASH-JSSH displayed the highest sensitivity to changes at three months, followed by VAS. All subscales of SF-36 were much less sensitive and both grip and pinch strength were unchanged over the three-month period. DASH-JSSH demonstrated more responsiveness to changes after carpal tunnel release than SF-36, VAS and physical measurements, and displayed correlations with subscales of SF-36. PMID- 17080526 TI - Aneurysmal bone cyst of the hand: a report of four cases. AB - Aneurysmal bone cysts are benign lesions that rarely occur in the bones of the hand. Curettage and bone grafting are the most common treatment modalities performed considering the possible functional loss after total excision. Four cases of aneurysmal bone cyst of the hand were identified. Three out of four cases were treated initially with curettage and bone grafting alone without any other local therapy. One had total excision of the lesion. There were two recurrences in the curettage group. Both were treated with excision and followed up for a minimum of 12 months. Curettage alone was associated with high recurrence rates. Total excision of the lesion was the most successful procedure employed. In view of the high recurrence rates following curettage alone and non neoplastic nature of the lesion, adjuvant treatment modalities such as cryosurgery, electric cauterisation should be considered for initial treatment of ABC. Resection should be preserved for recurrent cases. PMID- 17080527 TI - Acute rupture of extensor pollicis longus tendon due to avulsed fracture of the second metacarpal base: a case report. AB - Acute tendon rupture of the extensor pollicis longus (EPL) is rare. We present acute EPL tendon rupture associated with avulsed fracture of the second metacarpal at the insertion of extensor carpi radialis longus. Tendon rupture of the EPL was repaired with end-to-end suture and avulsed fracture of the second metacarpal base was treated with open reduction and internal fixation. PMID- 17080528 TI - An anomalous muscle mimicking a dorso-radial ganglion as a cause of radial wrist pain. AB - We report an abnormal muscle on the radial aspect of the wrist, which presented clinically as a ganglion and radial wrist pain. Existence of muscles in the first compartment has been accounted earlier, however its atypical presentation advocates its inclusion in the differential diagnosis of a dorso-radial ganglion and wrist pain. PMID- 17080529 TI - Digital papillary adenocarcinoma: a case report. AB - Digital papillary adenocarcinoma (DPAc) is a relatively rare neoplasm arising from the sweat glands with a predilection for the hand. A case of DPAc in the third finger at the level of the proximal phalanx in a 55-year-old male is presented. Our paper recommends specific consideration of DPAc in evaluating digital soft tissue masses, particularly those that present with an aggressive nature. PMID- 17080530 TI - Dynamic scapholunate dissociation in a child. AB - We describe a case of dynamic scapholunate dissociation in a skeletally immature ten-year-old girl. The patient presented for pain in the anatomical snuffbox and swelling over the dorsum of the wrist and hand. Standard static and dynamic radiographs of the wrist did not reveal any pathology. Magnetic resonance imaging was inconclusive. The scapholunate dissociation was diagnosed by wrist arthroscopy, reduced, and stabilised using standard techniques. Full and pain free motion of the wrist was restored. PMID- 17080531 TI - Locking of the metacarpophalangeal joint of the thumb by a loose body: a case report. AB - We report a case with locking of the metacarpophalangeal (MP) joint of the thumb in a 15-year-old high school baseball catcher, which was caused by an intra articular loose body arising from osteochondritis dissecans. The loose body was removed arthroscopically, enabling early return to full MP joint function. PMID- 17080532 TI - Simultaneous fractures of distal and proximal phalanges of the thumb, trapezium and carpal scaphoid of the ipsilateral hand: a case of very unusual combination of fractures. AB - Simultaneous multiple injuries of the thumb ray are rare. An unusual case of a simultaneous fracture of proximal and distal phalanges, trapezium and scaphoid is reported. The patient had an acceptable result six months after injury. This type of injury of the thumb ray may be a specific form of injury pattern caused by the mobile phone. PMID- 17080533 TI - Delayed rupture of flexor pollicis longus tendon after volar plating of the distal radius. AB - Flexor tendon rupture following distal radius fracture is rare. We described a case of flexor pollicis longus rupture, presented five years after volar plating of distal radius fracture. PMID- 17080534 TI - The effectiveness of RA wrist fusion using Beta-TCP without autogenous iliac bone grafting: a report of four cases. AB - Wrist arthrodesis is indicated for the rheumatoid hand especially in cases with severe destruction of the carpal bones. In the arthrodesis procedure for the rheumatoid wrist, autogenous iliac bone grafting is required in most cases. However, autogenous iliac bone graft necessitates the additional surgical intervention, and can be associated with the problem of inadequate bony quality or quantity. It is thought that use of the artificial bone substitute in the procedure can lessen the surgical morbidity while supplying the consistent material without shortage of graft quantity. We have performed arthrodesis of the rheumatoid wrist using beta-TCP for four patients. Clinical results of these patients were satisfactory both in pain relief and functional improvement with complete bony healing. Therefore, this procedure seems to be an effective option for the rheumatoid wrist with severe destructive changes. PMID- 17080535 TI - Necrotising fasciitis causing compartment syndrome of the forearm and septic shock due to Vibrio vulnificus: a case report. AB - Compartment syndrome caused by necrotising fasciitis has rarely been described. We report a case of systemic Vibrio vulnificus necrotising fasciitis presented with compartmental syndrome of the forearm and septic shock. The patient was treated with systemic antibiotic treatment and urgent surgical decompression followed by multiple necrotic tissue debridements. The patient recovered with some limited motion of the hand function. Prompt recognition and immediate treatment with antibiotics and surgical intervention are essential. PMID- 17080536 TI - Treating deep-seated mycobacterium marinum infection in the hand: a report of three cases. AB - We report three cases of an unusual aggressive type of Mycobacterium marinum infection of the hand which had been a nightmare both for us and the patient. These were the patients in which even after repeated thorough surgical debridement and appropriate (drug sensitive) medical treatment over a period had resulted in amputation or devastation of the soft tissues so extensively that almost only the neurovascular bundle, bone and the skin were the only structures left (cosmetic fingers - no function) at the last debridement after which the infection has not recurred. By presenting this article we want to stress on certain points pertaining to diagnosis, management, varied presentation and of course the dreadful complications of deep-seated M. marinum infection. PMID- 17080537 TI - Cubital tunnel syndrome due to giant cell tumour of tendon sheaths. AB - Cubital tunnel syndrome is one of the most common entrapment neuropathies in adults. It is mainly caused by the depression of ulnar nerve from normal structures at the elbow area. Despite the fact that several pathgological entities can be potential mechanisms of the syndrome, the pathogenesis due to benign or malignant neoplasms is extremely rare. In the present report we describe the first case of cubital tunnel syndrome due to giant cell tumour of the tendon sheaths. PMID- 17080538 TI - A comparative biomechanical study of the looped square slip knot and the simple surgical knot. AB - The looped square slip knot was introduced as a technique for skin closure to avoid the use of sharp instruments in suture removal after hand surgery. We compared the biomechanical properties of this knot with the simple surgical square knot. The ultimate strength of the looped square slip knot was significantly (p = 0.015) higher than the simple surgical knot. There was no significant difference between the two knots in mode of failure. Knot slippage or suture breakage did not occur in any samples when testing security by repetitive loading. Therefore, the looped square slip knot is a safe and convenient alternative to the two-throw surgical knot for use in hand surgery. PMID- 17080539 TI - Alternative therapies for osteoporosis. AB - A variety of common complementary and alternative medicine therapies are now being examined for effectiveness in the management of osteoporosis. Short-term studies in postmenopausal women show beneficial effects of soy isoflavone supplementation on bone density, but its long-term effects require clarification. Prospective controlled trials have shown that physical training can increase bone density to varying degrees. Other therapies that have been examined include herbal formulae, essential fatty acids and vitamins A, C, and K, but few data regarding their effectiveness, mechanisms and safety have been published. Further randomized controlled trials are needed. PMID- 17080541 TI - The effects of Baduanjin qigong in the prevention of bone loss for middle-aged women. AB - This study aimed to assess the efficacy of a 12-week Baduanjin qigong training program in preventing bone loss for middle-aged women. An experimental design was adopted, and subjects were assigned randomly into an experimental group (n = 44) and a control group (n = 43). The experimental group received a 12-week Baduanjin qigong training program, whereas the control group did not. Interleukin-6 (IL-6) and bone mineral density (BMD) were measured before and after the intervention. The results showed significant differences in IL-6 (t = -5.19, p < 0.000) and BMD (t = 1.99, p = 0.049) between the groups. Baduanjin reduced IL-6 and maintained BMD in the experimental group. In conclusion, this study demonstrates promising efficacy of Baduanjin in preventing bone loss commonly occurring in middle-aged women. Thus, Baduanjin is valuable for promoting and maintaining the health status of middle-aged women. PMID- 17080540 TI - Efficacy of the Kampo medicine Xiong-gui-jiao-ai-tang, a traditional herbal medicine, in the treatment of threatened abortion in early pregnancy. AB - This study was carried out to evaluate the clinical efficacy of Xiong-gui-jiao-ai tang (Kyuki-kyogai-to), a traditional Chinese herbal medicine, in the treatment of threatened abortion in early pregnancy. We enrolled 72 women diagnosed with threatened abortion at Osaka Medical College Hospital and assigned them at random to the following two groups: a group of 36 women who received Xiong-gui-jiao-ai tang at a dose of 7.5 g/day and another group of 36 women who received human chorionic gonadotropin (hCG)(control group). We found that in the Xiong-gui-jiao ai-tang group (2.9 + or - 3.5 days), the number of days required before hemostasis was reached in the uterus was significantly shorter than in the control group (10.8 + or - 8.2 days, p < 0.0001). Furthermore, the number of days required for retroplacental hematoma in the vicinity of the gestational sac to disappear was significantly shorter in the Xiong-gui-jiao-ai-tang group (9.9 + or - 7.1 days) than in the control group (23.2 + or - 12.8 days) (p < 0.0001). In retroplacental hematoma size, significant rates of reduction were obtained in both major and minor axis measurements at the 7th day of treatment for the Xiong gui-jiao-ai-tang group compared to the control group (control vs Xiong-gui-jiao ai-tang: major axis: 7.5 + or - 3.8% vs 42.3 + or - 10.5%; minor axis: 15.3 + or 16.8% vs 71.5 + or - 48.2%)(p < 0.0001, each case). The results of this study demonstrated the beneficial effects of Xiong-gui-jiao-ai-tang in stabilizing early pregnancy. Xiong-gui-jiao-ai-tang can be expected to improve unstable early pregnancy with uterine bleeding and to prevent abortion. PMID- 17080542 TI - A comparison between directional and proportional methods in locating acupuncture points using dual-energy X-ray absorptiometry in Korean women. AB - The cun measurement system, an essential and convenient method for locating acupoints, has been widely used in the practice of acupuncture. However, the traditional cun measurement methods have been criticized for their lack of reliability. In addition, new technology has become available to measure the soft tissue and bone mass independently. The present study was conducted in order to determine if one cun, measured by the directional method, is consistent with proportional methods and to investigate whether both methods can be used to locate acupoints. The lengths of bone from several parts of the body in 47 Korean women were measured using Dual-Energy X-ray Absorptiometry. The lengths of each part of the body were compared with 3 cun measured by proportional methods, and the width of all four fingers measured at the level of the proximal inter phalangeal joint used in the directional method. The results showed that one cun measured by the directional method was significantly different from one cun by proportional methods. The directional method is likely less dependable in locating acupoints than the proportional method. PMID- 17080543 TI - A study on the clinical effects of physical therapy and acupuncture to treat spontaneous frozen shoulder. AB - The integration of traditional Chinese and Western medicine and their clinical effects have been widely evaluated. Many studies have shown that using a combination of these two remedies has resulted in better outcomes than using only one of them. Acupuncture is a traditional Chinese medical technique, which plays an important role in enforcing pain control, prevention and functional improvement. In 1979, the World Health Organization (WHO) journal introduced acupuncture as a remedy for 43 diseases, including frozen shoulder. This study aims to assess the therapeutic outcomes of combining acupuncture and physical therapy to treat frozen shoulder, and hopes to establish an evidence-based study of the integration of acupuncture and western medicine in the future. A total of 75 frozen shoulder patients treated in a medical center were recruited for the study between January 2002 and December 2002. The average age of these patients was 54.8 years. The average duration of the condition was 25.8 weeks before treatment. Of the 75 patients, 30 were treated by physical therapy, 30 by acupuncture and 15 by both remedies. Before the treatment began, all patients were evaluated by assessing static pain scale, motion pain scale, active and passive ROM (range of motion) and quality of life scale sheet SF-36 (Short Form 36). The outcome was evaluated by follow-up assessments conducted at the 2nd week and 4th week of treatment sessions. All patients showed improvement in quality of life (Short Form-36). Pain was controlled better by acupuncture while ROM improved following physical therapy. However, patients treated by both methods had the best outcome. The integration of acupuncture and physical therapy to treat frozen shoulder leads to a better outcome than using only one method. The author suggests that an evidence-based foundation of the integration of Chinese and Western medicine should be established in the future, to encourage the integration of Chinese and Western medicine. PMID- 17080544 TI - Time course of changes in nail fold microcirculation induced by acupuncture stimulation at the Waiguan acupoints. AB - Acupuncture has been used widely to treat disease; however, the time course for acupuncture to have an effect remains unclear. Therefore, the aim of the present study was to investigate the time course of changes in nail fold microcirculation (NFM) induced by acupuncture stimulation (AS) at the right and left Waiguan acupoints (WAs). A total of 38 healthy female volunteers, age range from 21 to 33, were studied. We recorded NFM of the right middle finger before, and 5 min, 10 min, 15 min and 20 min after initiating AS; NFM was also recorded 5 min and 10 min after secessions of AS. Nitric oxide (NO) and endothelin-1 levels were measured from the left cubital vein, before AS and 10 min after stopping AS. The results indicated that capillary density of NFM increased 5 min after AS at the right Waiguan acupoint (WA); however, similar changes were not noted at the left WA. The capillary density decreased beginning 15 min after AS at the right and left WA. Capillary red blood cell velocity increased 5 min and 10 min after AS at the right and left WAs, but decreased 5 min and 10 min after stopping AS at the left WA. NO and endothein-1 levels were similar before AS and 10 min after stopping AS. Therefore, we suggest that a segmental effect of the spinal nerve contributes to the increasing capillary density of NFM induced by AS. The effect of acupuncture on NFM lasts about 10-15 min. The changes of balance between the sympathetic nerve activities and parasympathetic nerve activities may be induced by AS. PMID- 17080545 TI - Positron emission tomography analysis of the analgesic effects of acupuncture in rhesus monkeys. AB - The purpose of this study was to examine whether pain-induced brain activation was suppressed by acupuncture analgesia. We investigated the suppression of the pain-induced neuronal activation in specific brain areas of three male rhesus monkeys (aged four years old) using positron emission tomography (PET), in which changes in the regional cerebral blood flow (rCBF) were examined as an index of the neuronal activation. The brain areas such as the thalamus, insula and anterior cingulate cortex were activated by heating the tail of monkeys in 47 degrees C water compared to the heating at 37 degrees C. Those activations were suppressed by electroacupuncture (EA) with a 2 sec alteration of the frequency of 4 Hz/60 Hz at the right ST36 (the upper anterior tibial muscle) and the right LI4 (the back palm between the first and second metacarpal) acupoints. Meanwhile, this EA analgesic effect was confirmed by prolonging the tail withdrawal latencies from hot water in the temperature range from 45 to 50 degrees C.These brain areas were corresponded to the pain-related areas as reported in previous studies. In conclusion, we were able to visualize the acupuncture analgesia in the CNS. We also detected the brain areas activated or inactivated by acupuncture. The areas that responded to acupuncture stimulation at 47 degrees C water were different from the regions at 37 degrees C. We consider that this difference in the response to acupuncture may support the variation of the clinical efficacy of acupuncture in patients bearing pain or other disorders. PMID- 17080546 TI - Ginkgo biloba leaf extract (EGb761) combined with neuroprotective agents reduces the infarct volumes of gerbil ischemic brain. AB - Ginkgo biloba exerts many pharmacological actions. It possesses antioxidant properties, the ability of neurotransmitter/receptor modulation and antiplatelet activation factor. This research is designed to investigate the neuroprotective effects of long-term treatment with EGb761 (a standard form of the extract of Ginkgo biloba leaf) in combination with MgSO(4), FK506, or MK-801 on the infarct volume of male gerbils' brain induced by unilateral middle cerebral artery occlusion (MCAO). Thirty-five gerbils fed a standard diet were intragastrically given water or EGb761 (100 mg/kg/day) for one week. Five randomized groups were established: control (n = 7), EGb761 (n = 8), EGb761 + MgSO(4) (n = 7), EGb761 + FK506 (n = 7), and EGb761 + MK-801 (n = 6). The three drug-combination groups were injected with MgSO(4) (90 mg/kg), FK506 (0.5 mg/kg), or MK-801 (1 mg/kg), respectively 30 min before MCAO. Gerbils were anesthetized and craniectomized to expose the right middle cerebral artery (MCA). The right MCA was constricted with an 8-0 suture to produce a permanent ligation for 24 hours. Postmortem infarct volumes were determined by quantitative image analysis of 2,3,5 triphenyltetrazolium chloride (TTC)-stained brain sections. Results showed that the total infarct volumes of the four treated groups either EGb761 alone or in combination with drugs were lower than the control group by 36.1% (EGb761 alone), 40.3% (EGb761 + MgSO(4)), 35.3% (EGb761 + FK506), and 56.4% (EGb761 + MK-801), respectively (p < 0.01). The main affected areas of the brain in the four treated groups were significantly focused between 4 and 6 mm from the frontal pole, when compared to the control group (p < 0.01). All animals in the five groups had infarctions in both cortex and subcortex. These results indicate that long-term pre-treatment of EGb761 administered either alone or in combination with drugs significantly effective neuroprotection on infarct volume in gerbil ischemic brains. PMID- 17080547 TI - Anti-hyperglycemic activity of natural and fermented Cordyceps sinensis in rats with diabetes induced by nicotinamide and streptozotocin. AB - Our previous study demonstrated that the fruiting bodies of Cordyceps sinensis, a traditional Chinese medicine, attenuated diabetes-induced weight loss, polydipsia, and hyperglycemia in rats. In the present study, we further compared the anti-hyperglycemic activity of the fermented mycelia and broth of Cordyceps sinensis with that of the fruiting bodies. Male Wistar rats orally administered a placebo (STZ group), fruiting bodies (FB group, 1 g/day), fermented mycelia (MCS group, 1 g/day), fermented broth (BCS group, 1 g/day), or fermented mycelia plus broth (XCS group, 0.5 g/day of each) of Cordyceps sinensis (d1 to d28) were injected with nicotinamide (200 mg/kg) and streptozotocin (65 mg/kg) on d15. Rats fed with a placebo and injected with saline served as the control (CON) group. The amount of water and food consumption (d15 to d29), the 2-hour-postprandial blood glucose concentrations (d21 and d28), and the serum concentrations of fructosamine (d29) were significantly lower in the FB, MCS, BCS, and XCS groups than in the STZ group (one-way ANOVA, p < 0.05). The diabetic rats had significantly higher blood glucose concentrations as measured by the oral glucose tolerance test than the control rats; moreover, these changes were significantly reduced by ingesting the fruiting bodies, fermented mycelia and/or broth of Cordyceps sinensis. Our results revealed that the fermented mycelia and broth of Cordyceps sinensis have anti-hyperglycemic activities similar to those of the fruiting bodies. Therefore, the fermented products of Cordyceps sinensis could be developed as potential anti-diabetic agents or functional foods for persons with a high risk of diabetes mellitus. PMID- 17080548 TI - Anti-hyperalgesic and anti-inflammatory effects of the modified Chinese herbal formula Huo Luo Xiao Ling Dan (HLXL) in rats. AB - Chinese herbal medicine has been used for thousands of years in China and other Asian countries to treat a variety of inflammatory diseases. The classic Chinese herbal formula, Huo Luo Xiao Ling Dan (HLXL) is commonly used in traditional Chinese herbal medicine for the treatment of joint pain and other symptoms of arthritis. The present study is an investigation of the effects of a modified HLXL extract on persistent hyperalgesia and edema in rats with peripheral inflammation. Inflammation was induced by injecting complete Freund's adjuvant (CFA) into one hind paw. Four dosages of the extract were compared to a vehicle control. Each was administered intragastrally (i.g.) daily for seven days beginning one day before CFA. Hyperalgesia was assessed using a paw withdrawal latency (PWL) test and edema was determined by measuring paw thickness at pre-CFA and 2 hours, 24 hours, and 5 days post-CFA. Immunohistochemistry was performed 2 hours post-CFA to determine spinal Fos protein expression. Adverse effects of the extract were monitored by observing the animals closely for unusual behavioral changes. Compared to the control, HLXL at the two lower dosages (0.575 g/kg and 1.15 g/kg) were effective in the later stage (day 5) of inflammatory hyperalgesia and edema, while the two higher dosages (2.3 g/kg and 4.6 g/kg) alleviated early stage hind paw inflammation and hyperalgesia and facilitated recovery from paw edema and hyperalgesia during the late stage. HLXL at 2.30 g/kg significantly suppressed Fos expression in laminae I-II, III-IV and V-VI ipsilaterally and in III-IV contralaterally. No significant signs of toxicity or adverse effects were observed. The data suggest that HLXL dosage-dependently attenuates CFA-induced inflammation and hyperalgesia, at least in part by inhibiting noxious transmission at the dorsal horn of the spinal cord. PMID- 17080549 TI - Ginger significantly decreased the oral bioavailability of cyclosporine in rats. AB - Ginger (roots of Zingiber officinale ROSCOE) is a popular spice and herbal medicine worldwide. Cyclosporine is clinically used as an important immunosupressant with narrow therapeutic index. This study attempted to investigate the effect of ginger juice on the pharmacokinetics of cyclosporine in rats. Rats were orally administered cyclosporine alone and in combination with ginger juice (5 ml/kg) concomitantly, as well as 2 hours after the ginger juice, respectively, in crossover designs. In addition, rats were intravenously administered cyclosporine with and without an oral dose of ginger juice (5 ml/kg). The blood samples were withdrawn via cardiopuncture at determined time points and cyclosporine concentrations were determined by a specific monoclonal fluorescence polarization immunoassay. The pharmacokinetic parameters of cyclosporine were calculated using a non-compartment model of WINNONLIN. The results indicated that concomitant intake of ginger significantly decreased C(max) and AUC(0-t) of oral cyclosporine by 70.9% and 63.1%, respectively. The intake of ginger 2 hours before cyclosporine significantly decreased C(max) and AUC(0-t) by 51.4% and 40.3%, respectively. In contrast, the pharmacokinetics of intravenous cyclosporine not altered by orally in combination with ginger juice. In conclusion, ginger significantly decreased the oral bioavailability of cyclosporine, and the interaction should occur at the absorption phase. Patients treated with cyclosporine should be discouraged from using ginger products to ensure the efficacy of cyclosporine. PMID- 17080550 TI - Activation of nitric oxide signaling pathway mediates hypotensive effect of Muntingia calabura L. (Tiliaceae) leaf extract. AB - The cardiovascular effect of the crude methanol extract from the leaf of Muntingia calabura L. (Tiliaceae) was investigated in the anesthetized rats. The crude methanol extract was sequentially fractionated to obtain the water-soluble extract (WSE). Intravenous administration of the WSE (10, 25, 50, 75 or 100 mg/kg) produced an initial followed by a delayed decrease in systemic arterial pressure (SAP) in a dose-dependent manner. The M. calabura-induced initial hypotension lasted for 10 min and the delayed depressor effect commenced after 90 min and lasted for at least 180 min post-injection. The same treatment, on the other hand, had no appreciable effect on heart rate (HR) or the blood gas/electrolytes concentrations. Both the initial and delayed hypotensive effects of WSE (50 mg/kg, i.v.) were significantly blocked by pre-treatment with a nonselective nitric oxide (NO) synthase (NOS) inhibitor, N(G)-nitro-L-arginine methyl ester ((L)-NAME, 0.325 mg/kg/min for 5 min) or a soluble guanylate cyclase (sGC) inhibitor, 1H-[1,2,4]oxadiazole[4,3-alpha]quinoxalin-1-one (ODQ, 0.2 mg/kg/min for 5 min). Moreover, whereas the initial depressor effect of WSE was inhibited by pre-treatment with a selective endothelial NOS (eNOS) inhibitor, N5 (1-Iminoethyl)-L-ornithine ((L)-NIO, 1 mg/kg/min for 5 min), the delayed hypotension was attenuated by a selective inducible NOS (iNOS) inhibitor, S methylisothiourea (SMT, 0.5 mg/kg/min for 5 min). Administration of WSE also produced an elevation in plasma nitrate/nitrite concentration, as well as an increase in the expression of iNOS protein in the heart and thoracic aorta. These results indicate that WSE from the leaf of M. calabura elicited both a transient and delayed hypotensive effect via the production of NO. Furthermore, activation of NO/sGC/cGMP signaling pathway may mediate the M. calabura-induced hypotension. PMID- 17080551 TI - Osteogenic potential using a malleable, biodegradable composite added traditional Chinese medicine: in vitro and in vivo evaluations. AB - The purpose of this investigation was to prepare and evaluate the feasibility and biocompatibility of a new composite as a large defect bone substitute. The new GTGG was mainly composed of tricalcium phosphate ceramic particles and glutaraldehyde crosslinked gelatin in which Gui-Lu-Jiao was added (a mixture of Cervi Colla Cornus and Colla Plastri Testudinis). In the in vitro study, rat's calvaria osteoblasts were used to study bone characteristics upon exposure to different concentrations of the Gui-Lu-Jiao solution. In the in vivo study, GTGG composites were implanted into the defects of calvarial bones in mature New Zealand rabbits to test their osteogenerative characteristics. As a result, we found that Gui-Lu-Jiao added to the culture could promote the proliferation of osteoblasts. In addition, GTGG could induce a large amount of new bone growth in the rabbit's calvarial bone defect. Therefore, the GTGG composite might be a potential bone substitute. PMID- 17080552 TI - Mechanisms of pseudolaric acid B-induced apoptosis in Bel-7402 cell lines. AB - Previous studies have shown that pseudolaric acid B (PB) would cause apoptosis in human tumor cell lines. However, the mechanisms of PB induced apoptosis are still unclear. In the present study, the mechanisms of PB induced apoptosis in the human hepatocellular carcinoma Bel-7402 cell line were investigated by measuring cell viability, rate of apoptosis, cell cycle, detecting DNA fragmentation, and measuring caspase-3 activation. The results indicated that PB inhibited Bel-7402 cell viability and induced cell death by causing DNA fragmentation, up regulating the early and late apoptotic rates, activating caspase-3 protein, and detaining the cell cycle in the G2/M phases. Additionally, PB-induced apoptosis was a dose- and time-dependent manner. These observations suggest that PB-induced apoptosis occurs through a caspase-dependent pathway and detains the cell cycle in the G2/M phase. PMID- 17080553 TI - The effects of five alkaloids from Bulbus Fritillariae on the concentration of cAMP in HEK cells transfected with muscarinic M(2) receptor plasmid. AB - The aim of this study was to investigate the effects of five alkaloids, namely verticine, verticinone, imperialine, imperialine-3beta-D-glucoside, and puqietinone, purified from Bulbus Fritillariae and used as an antitussive drug in traditional Chinese medicine, on their antimuscarinic M(2) function and the cAMP level of HEK cells transfected with muscarinic M(2) receptor plasmid. By transfecting the HEK cells with the method of calcium phosphate co-precipitation and screening with G418, the cells stably expressing M(2) receptor were identified. The expression of M(2) receptor in HEK cells was confirmed by both RT PCR and western blot. The cAMP level in the treated cells was analyzed with RIA method ((125)I-cAMP KIT). And the results suggested that the five alkaloids could significantly elevate the cAMP concentration in the HEK cells transfected with muscarinic M(2) receptor plasmid (p < 0.01). PMID- 17080554 TI - Effect of epigallocatechingallate on ultraviolet B-induced photo-damage in keratinocyte cell line. AB - One type of traditional Chinese medicines, epigallocatechingallate (EGCG) has been commonly used as a clinical and skin health protective ingredient. It has been known to have photo-protective, anti-inflammatory, and anti-oxidant effects. However, little is known about the mechanisms of EGCG on UV-induced photo-aging and photo-carcinogenesis. In the present study, we investigated the photo protective mechanisms of EGCG on UVB-induced skin damage, including the potency of EGCG to inhibit the UVB-induced cytotoxicity, secretion of cytokine (IL-6 and TNF-alpha), cellular apoptosis, expression of apoptosis-regulatory genes (p53 p21) and c-fos gene in cultured immortalized human keratinocyte HaCaT cells. EGCG treatment decreased UVB- induced cell cytotoxicity and apoptosis. It also inhibited the mRNA expressions of apoptosis-regulatory gene (p53 and p21) and c fos gene. These results suggest that EGCG may have an inhibitory effect on UVB induced photo-damage and apoptosis by blocking the cytokine secretion and the mRNA expressions of p53, p21 and c-fos genes. PMID- 17080555 TI - [Native pulmonary and aortic value endocarditis by Streptococcus bovis]. PMID- 17080556 TI - Re: cost analysis of neonatal circumcision in a large health maintenance organization. PMID- 17080557 TI - Re: cost analysis of neonatal circumcision in a large health maintenance organization. PMID- 17080558 TI - Re: cost analysis of neonatal circumcision in a large health maintenance organization. PMID- 17080559 TI - [Internist mission at the modern hospital]. PMID- 17080560 TI - Origins of ecstasy an urban myth. PMID- 17080561 TI - Final diagnosis: solitary fibrous tumor of the kidney. PMID- 17080562 TI - [NSAIDs and cardiovascular and renal complications]. PMID- 17080563 TI - Limitations of impedance cardiography. PMID- 17080564 TI - Challenges of vaccination. PMID- 17080565 TI - [Dubious documentation of the effect of acupuncture]. PMID- 17080566 TI - [Acute meningoencephalitis due to type 1 herpes virus simple]. PMID- 17080567 TI - Wanted: a few good sperm (looking for Mr. Good Sperm). PMID- 17080568 TI - Looking for the lie: scientists are using brain imaging and other tools as new kinds of lie detectors. But trickier even than finding the source of deception might be navigating a world without it. PMID- 17080569 TI - My father's abortion war: when the abortion provider Dr. Barnett Slepian was murdered outside Buffalo, one of his colleagues decided to keep his own nearby office open. How America's most polarizing conflict came to my family's doorstep. PMID- 17080570 TI - What would a clone say? A humanist case against therapeutic cloning. PMID- 17080571 TI - Will we ever arrive at the good death? Almost 40 years after the birth of the hospice movement, and despite the rise of living wills and palliative care, the end of life remains anxious and hypermedicalized. Goldie Gold's struggle, and ours, to come to a dignified end. PMID- 17080572 TI - The mystery of my eggs. PMID- 17080573 TI - Preschool meds: the first clinical trial examining the effects of generic Ritalin on 3- to 5-year-old subjects raises questions not only about the safety of the drug but also about the ethics of testing on ever younger brains. PMID- 17080574 TI - [Streptococcus Viridans emboligen endocarditis]. PMID- 17080575 TI - [Topiramate in migraine prophylaxis]. PMID- 17080576 TI - No increase in CNS defects. PMID- 17080577 TI - Growth hormone impacts thymic output. PMID- 17080578 TI - A new era. PMID- 17080579 TI - Fluoroquinolone-resistant Salmonella sp. in carcasses. PMID- 17080580 TI - Cocirculation of dengue serotypes, Delhi, India, 2003. PMID- 17080581 TI - Borrelia burgdorferi and Anaplasma phagocytophilum coinfection. PMID- 17080582 TI - Corynebacterium pseudogenitalium urinary tract infection. PMID- 17080583 TI - Puumala virus RNA in patient with multiorgan failure. PMID- 17080584 TI - The ins and outs of medication delivered by enteral tube. PMID- 17080585 TI - Reptile-associated salmonellosis in man, Italy. PMID- 17080586 TI - Congenital visceral leishmaniasis. PMID- 17080587 TI - Endometrial safety, overall safety and tolerability of transdermal continuous combined hormone replacement therapy over 96 weeks: a randomized open-label study. AB - OBJECTIVES: To establish whether transdermal continuous hormone replacement therapy (HRT) with estrogen/progestogen provides adequate long-term endometrial protection in postmenopausal women over a period of 96 weeks. METHODS: This multicenter, randomized, open-label, parallel-group study evaluated the endometrial effects and overall safety and tolerability of a transdermal matrix patch delivering estradiol (E2) 50 microg/day and norethisterone acetate (NETA) 140 microg/day (Estalis; patches applied twice weekly without intermediate breaks) and a once-daily oral comparator (Kliogest; one tablet containing E2 2 mg/NETA 1 mg) in postmenopausal women. A total of 406 women with an intact uterus, aged 44-69 years, were randomized in the 48-week core phase of the study, and 239 continued into the 48-week extension phase. Subjects were randomized in the ratio 3 : 1 to transdermal or oral E2/NETA treatment. RESULTS: No cases of endometrial hyperplasia or endometrial cancer were reported with either treatment during the core or extension phase. Both treatments were generally well tolerated, with most adverse events (>90%) being mild to moderate, although minor differences in the tolerability profile were observed between treatments. CONCLUSIONS: Continuous combined transdermal HRT with E2/NETA shows no evidence of an increased endometrial hyperplasia or endometrial cancer risk over a 96-week period. PMID- 17080588 TI - Can improvement in photosynthesis increase crop yields? AB - The yield potential (Yp) of a grain crop is the seed mass per unit ground area obtained under optimum growing conditions without weeds, pests and diseases. It is determined by the product of the available light energy and by the genetically determined properties: efficiency of light capture (epsilon i), the efficiency of conversion of the intercepted light into biomass (epsilon c) and the proportion of biomass partitioned into grain (eta). Plant breeding brings eta7 and epsilon i close to their theoretical maxima, leaving epsilon c, primarily determined by photosynthesis, as the only remaining major prospect for improving Yp. Leaf photosynthetic rate, however, is poorly correlated with yield when different genotypes of a crop species are compared. This led to the viewpoint that improvement of leaf photosynthesis has little value for improving Yp. By contrast, the many recent experiments that compare the growth of a genotype in current and future projected elevated [CO2] environments show that increase in leaf photosynthesis is closely associated with similar increases in yield. Are there opportunities to achieve similar increases by genetic manipulation? Six potential routes of increasing epsilon c by improving photosynthetic efficiency were explored, ranging from altered canopy architecture to improved regeneration of the acceptor molecule for CO2. Collectively, these changes could improve epsilon c and, therefore, Y p by c. 50%. Because some changes could be achieved by transgenic technology, the time of the development of commercial cultivars could be considerably less than by conventional breeding and potentially, within 10-15 years. PMID- 17080589 TI - Transgenic approaches to manipulate the environmental responses of the C3 carbon fixation cycle. AB - The limitation to photosynthetic CO2 assimilation in C3 plants in hot, dry environments is dominated by ribulose 1.5-bisphosphate carboxylase/oxygenase (Rubisco) because CO2 availability is restricted and photorespiration is stimulated. Using a combination of genetic engineering and transgenic technology, three approaches to reduce photorespiration have been taken; two of these focused on increasing the carboxylation efficiency of Rubisco either by reducing the oxygenase reaction directly or by manipulating the Rubisco enzyme by concentrating CO2 in the region of Rubisco through the introduction of enzymes of the C4 pathway. The third approach attempted to reduce photorespiration directly by manipulation of enzymes in this pathway. The progress in each of these areas is discussed, and the most promising approaches are highlighted. Under saturating CO2 conditions, Rubisco did not limit photosynthesis, and limitation shifted to ribulose bisphosphate (RuBP) regeneration capacity of the C3 cycle. Transgenic analysis was used to identify the specific enzymes that may be targets for improving carbon fixation, and the way this may be exploited in the high CO2 future is considered. PMID- 17080590 TI - Functional dynamics of plant growth and photosynthesis--from steady-state to dynamics--from homogeneity to heterogeneity. AB - Plants are much more dynamic than we usually expect them to be. This dynamic behaviour is of paramount importance for their performance under natural conditions, when resources are distributed heterogeneously in space and time. However, plants are not only the cue ball of their physical and chemical environment. Endogenous rhythms and networks controlling photosynthesis and growth buffer plant processes from external fluctuations. This review highlights recent evidence of the importance of dynamic temporal and spatial organization of photosynthesis and of growth in leaves and roots. These central processes for plant performance differ strongly in their dependence on environmental impact and endogenous properties, respectively. Growth involves a wealth of processes ranging from the supply of resources from external and internal sources to the growth processes themselves. In contrast, photosynthesis can only take place when light and CO2 are present and thus clearly requires 'input from the environment'. Nevertheless, growth and photosynthesis are connected to each other via mechanisms that are still not fully understood. Recent advances in imaging technology have provided new insights into the dynamics of plant-environment interactions. Such processes do not only play a crucial role in understanding stress response of plants under extreme environmental conditions. Dynamics of plants under modest growth conditions rise from endogenous mechanisms as well as exogenous impact too. It is thus an important task for future research to identify how dynamic external conditions interact with plant-internal signalling networks to optimize plant behaviour in real time and to understand how plants have adapted to characteristic spatial and temporal properties of the resources from their environment, on which they depend on. PMID- 17080591 TI - The importance of maltose in transitory starch breakdown. AB - Starch content of leaves responds to environmental stresses in various ways. Understanding these environmental effects on starch metabolism has been difficult in the past because the pathways of transitory starch synthesis and degradation are not completely known. Over the past two years there has been a significant increase in our understanding of transitory starch breakdown. The discovery of a maltose transporter (MEX1) and the studies of a cytosolic disproportionating enzyme (D-enzyme, DPE2) confirmed that maltose is the predominant form of carbon exported from chloroplasts at night. Maltose increases in leaves when starch breakdown is induced during the day under photorespiratory conditions. Maltose metabolism is regulated by a circadian clock, day length and temperature. The expression of maltose-metabolizing genes shows a pronounced circadian rhythm indicating maltose metabolism is clock regulated. Indeed, the maltose level oscillates under continuous light. The transcript of a beta-amylase gene (BAM3) peaks during the day in long days and peaks at night in short days. This could provide a mechanism for adjusting starch breakdown rates to day length. Under cold-stress conditions, maltose increases and BAM3 expression is induced. We hypothesize that maltose metabolism is a bridge between transitory starch breakdown and the plants' adaptation to changes in environmental conditions. PMID- 17080592 TI - The hydraulic limitation hypothesis revisited. AB - We proposed the hydraulic limitation hypothesis (HLH) as a mechanism to explain universal patterns in tree height, and tree and stand biomass growth: height growth slows down as trees grow taller, maximum height is lower for trees of the same species on resource-poor sites and annual wood production declines after canopy closure for even-aged forests. Our review of 51 studies that measured one or more of the components necessary for testing the hypothesis showed that taller trees differ physiologically from shorter, younger trees. Stomatal conductance to water vapour (g(s)), photosynthesis (A) and leaf-specific hydraulic conductance (K L) are often, but not always, lower in taller trees. Additionally, leaf mass per area is often greater in taller trees, and leaf area:sapwood area ratio changes with tree height. We conclude that hydraulic limitation of gas exchange with increasing tree size is common, but not universal. Where hydraulic limitations to A do occur, no evidence supports the original expectation that hydraulic limitation of carbon assimilation is sufficient to explain observed declines in wood production. Any limit to height or height growth does not appear to be related to the so-called age-related decline in wood production of forests after canopy closure. Future work on this problem should explicitly link leaf or canopy gas exchange with tree and stand growth, and consider a more fundamental assumption: whether tree biomass growth is limited by carbon availability. PMID- 17080593 TI - Managing sulphur metabolism in plants. AB - Resolution and analysis of genes encoding components of the pathways of primary sulphur assimilation have provided the potential to elucidate how sulphur is managed by plants. Individual roles for members of gene families and regulatory mechanisms operating at gene, cellular and whole plant levels have been recognized. Sulphur is taken up and transported around the plant principally as sulphate, catalysed for the most part by a single gene family of highly regulated transporters. Additional regulation occurs in the pathway of reduction of sulphate to sulphide and its incorporation into cysteine, which occurs principally within the plastid. Cellular and whole-plant regulation of uptake, and the assimilatory pathway attempt to balance supply with demand for growth and include mechanisms for re-mobilization and redistribution of sulphur. Furthermore, optimization of sulphur assimilation requires coordination with carbon and nitrogen pathways, and multiple processes have been proposed to contribute to this balance. Present studies on cis and trans elements are focusing on transcriptional regulation, but this regulation still needs to be linked to apparent metabolite sensing. Whilst the components of the assimilatory pathways have been resolved after many years of controversy, uncertainties remain concerning roles of individual genes in gene families, their sub-cellular localization and their significance in balancing sulphur flux to sulphur demand of the plant for growth under variable environmental conditions. PMID- 17080594 TI - Diurnal regulation of plant growth. AB - Life occurs in an ever-changing environment. Some of the most striking and predictable changes are the daily rhythms of light and temperature. To cope with these rhythmic changes, plants use an endogenous circadian clock to adjust their growth and physiology to anticipate daily environmental changes. Most studies of circadian functions in plants have been performed under continuous conditions. However, in the natural environment, diurnal outputs result from complex interactions of endogenous circadian rhythms and external cues. Accumulated studies using the hypocotyl as a model for plant growth have shown that both light signalling and circadian clock mutants have growth defects, suggesting strong interactions between hypocotyl elongation, light signalling and the circadian clock. Here, we review evidence suggesting that light, plant hormones and the circadian clock all interact to control diurnal patterns of plant growth. PMID- 17080595 TI - Metabolic signalling in defence and stress: the central roles of soluble redox couples. AB - Plant growth and development are driven by electron transfer reactions. Modifications of redox components are both monitored and induced by cells, and are integral to responses to environmental change. Key redox compounds in the soluble phase of the cell are NAD, NADP, glutathione and ascorbate--all of which interact strongly with reactive oxygen. This review takes an integrated view of the NAD(P)-glutathione-ascorbate network. These compounds are considered not as one-dimensional 'reductants' or 'antioxidants' but as redox couples that can act together to condition cellular redox tone or that can act independently to transmit specific information that tunes signalling pathways. Emphasis is placed on recent developments highlighting the complexity of redox-dependent defence reactions, and the importance of interactions between the reduction state of soluble redox couples and their concentration in mediating dynamic signalling in response to stress. Signalling roles are assessed within the context of interactions with reactive oxygen, phytohormones and calcium, and the biochemical reactions through which redox couples could be sensed are discussed. PMID- 17080596 TI - Sugar and ABA response pathways and the control of gene expression. AB - Sugars are essential to plant growth and metabolism, both as energy source and as structural components. Sugar production and use are in part controlled at the level of gene expression by the sugars themselves. Responses to sugar are closely integrated with response pathways that indicate environmental conditions such as light and water availability. High sugar levels inhibit seedling development, repress photosynthetic gene expression and induce genes of storage metabolism such as those of starch biosynthesis. Genetic approaches have demonstrated the importance of abscisic acid (ABA) and the transcriptional regulator ABA insensitive4 (ABI4) in sugar response pathways. Recent analysis of both photosynthetic and starch biosynthetic gene promoters suggest a direct role for ABI4 in their control. The increased understanding of the regulatory promoter elements controlling gene expression, in response to sugar and ABA, allows transcriptional networks to be understood at a molecular level. PMID- 17080597 TI - Carotenoid accumulation and function in seeds and non-green tissues. AB - Carotenoids are plant pigments that function as antioxidants, hormone precursors, colourants and essential components of the photosynthetic apparatus. Carotenoids accumulate in nearly all types of plastids, not just the chloroplast, and are thus found in most plant organs and tissues, albeit at trace levels in some tissues. In this review we summarise the current knowledge of the carotenoid content of non-green plastids and discuss what is known about the regulation of their biosynthesis in roots, fruits, flowers, tubers and seeds. The emphasis is on food crops as carotenoids are essential components of human diets, primarily as some are precursors of vitamin A. The low carotenoid content of many staple foods, such as cereals, can exacerbate dietary deficiencies. The World Health Organisation has estimated that more than 100 million children are vitamin A deficient and up to 500,000 of these children become blind each year. Many of these children die within 12 months of going blind. Thus, understanding the regulation of carotenoid accumulation in food crops, especially tubers and cereals, should facilitate improvements to nutritional value with potentially significant health benefits. PMID- 17080598 TI - Brassinosteroids and plant function: some clues, more puzzles. AB - The role of brassinosteroids (BRs) in plant function has been intensively studied in the last few years. Mutant analysis has demonstrated that the ability to synthesize, perceive and respond to BRs is essential to normal plant growth and development. Several key elements of BR response have been identified using both genetic and biochemical approaches, and molecular models that parallel Wingless (Wnt), transforming growth factor beta (TGF beta) and receptor tyrosine kinase (RTK) signalling in animals have been proposed. Many studies have demonstrated the role of BRs, alone and in interaction with other plant hormones, in processes such as cell elongation and seed germination. In contrast, little is known about how the sensing of BRs is connected to specific physiological responses such as stress resistance. There remain many open questions about how these connections are made. PMID- 17080599 TI - Oxidative gating of water channels (aquaporins) in corn roots. AB - An oxidative gating of water channels (aquaporins: AQPs) was observed in roots of corn seedlings as already found for the green alga Chara corallina. In the presence of 35 mM hydrogen peroxide (H2O2)--a precursor of hydroxyl radicals (*OH)--half times of water flow (as measured with the aid of pressure probes) increased at the level of both entire roots and individual cortical cells by factors of three and nine, respectively. This indicated decreases in the hydrostatic hydraulic conductivity of roots (Lp(hr)) and of cells (Lp(h)) by the same factors. Unlike other stresses, the plant hormone abscisic acid (ABA) had no ameliorative effect either on root LP(hr) or on cell Lp(h) when AQPs were inhibited by oxidative stress. Closure of AQPs reduced the permeability of acetone by factors of two in roots and 1.5 in cells. This indicated that AQPs were not ideally selective for water but allowed the passage of the organic solute acetone. In the presence of H2O2, channel closure caused anomalous (negative) osmosis at both the root and the cell level. This was interpreted by the fact that in the case of the rapidly permeating solute acetone, channel closure caused the solute to move faster than the water and the reflection coefficient (sigma s) reversed its sign. When H2O2 was removed from the medium, the effects were reversible, again at both the root and the cell level. The results provide evidence of oxidative gating of AQPs, which leads on to inhibition of water uptake by the roots. Possible mechanisms of the oxidative gating of AQPs induced by H2O2 (*OH) are discussed. PMID- 17080600 TI - NMR metabolite profiling analysis reveals changes in phospholipid metabolism associated with the re-establishment of desiccation tolerance upon osmotic stress in germinated radicles of cucumber. AB - The adaptation of metabolism is thought to play a role in the acquisition of desiccation tolerance (DT). However, the importance of such a role and whether specific regulatory pathways exist remain to be assessed. Using in vitro 31P and 13C nuclear magnetic resonance (NMR) spectroscopy and biochemical assays, we analysed metabolite profiles of perchloric extracts from germinating radicles of cucumber to identify changes in carbon and phosphate metabolism associated with DT. Emerged radicles measuring 2 mm long can be rendered tolerant to desiccation by incubation in a polyethylene glycol (PEG) solution with a water potential of 1.5 MPa. However, in 4-mm-long emerged radicles, this treatment was ineffective. This manipulable system enabled the discrimination of changes in metabolites associated with DT from those associated with the response to osmotic stress. Independent of radicle length, the PEG treatment resulted in an increase in sucrose (Suc) content, whereas glucose (Glc), fructose (Fru) and the hexose phosphate pool, as well as phosphoenolpyruvate decreased three- to fourfold. In addition, three derivatives arising early during phospholipid catabolism (glycerylphosphorylcholine, glycerylphosphorylethanolamine and glycerylphosphorylinositol) appeared in the PEG-treated radicles. Interestingly, phospholipid degradation was much more pronounced in osmotically challenged radicles that remain sensitive to drying. This was proved by the appearance of catabolites, such as phosphocholine and phosphoethanolamine, solely in 4 mm PEG treated radicles. Furthermore, glycerol-3-phosphate and its derivative 3 phosphoglycerate increased significantly. Our data suggest that the metabolic response leading to the re-establishment of DT is not entirely identical to that of an osmotic response. It is inferred that membrane remodelling and/or increased phospholipid catabolism is an adaptive response common to osmotic adjustment and DT but is controlled differently in tolerant and sensitive radicles. PMID- 17080601 TI - Using continuous stable isotope measurements to partition net ecosystem CO2 exchange. AB - Ecosystem-scale estimation of photosynthesis and respiration using micrometeorological techniques remains an important, yet difficult, challenge. In this study, we combined micrometeorological and stable isotope methods to partition net ecosystem CO2 exchange (FN) into photosynthesis (F(A)) and respiration (F(R)) in a corn-soybean rotation ecosystem during the summer 2003 corn phase. Mixing ratios of (12)CO2 and (13)CO2 were measured continuously using tunable diode laser (TDL) absorption spectroscopy. The dynamics of the isotope ratio of ecosystem respiration (R), net ecosystem CO2 exchange (deltaN) and photosynthetic discrimination at the canopy scale (delta canopy) were examined. During the period of full canopy closure, F(N) was partitioned into photosynthesis and respiration using both the isotopic approach and the conventional night-time-derived regression methodology. Results showed that deltaR had significant seasonal variation (-32 to -11% per hundred) corresponding closely with canopy phenology. Daytime deltaN typically varied from -12 to -4% per hundred, while delta canopy remained relatively constant in the vicinity of 3% per hundred. Compared with the regression approach, the isotopic flux partitioning showed more short-term variations and was considerably more symmetric about F(N). In this experiment, the isotopic partitioning resulted in larger uncertainties, most of which were caused by the uncertainties in deltaN. and the daytime estimate of deltaR. By sufficiently reducing these uncertainties, the tunable diode laser (TDL)-micrometeorological technique should yield a better understanding of the processes controlling photosynthesis, respiration and ecosystem-scale discrimination. PMID- 17080602 TI - Spatial and temporal scaling of intercellular CO2 concentration in a temperate rain forest dominated by Dacrydium cupressinum in New Zealand. AB - Seven methods, including measurements of photosynthesis (A) and stomatal conductance (g(s)), carbon isotope discrimination, ecosystem CO2 and water vapour exchange using eddy covariance and the use of a multilayer canopy model and ecosystem Keeling plots, were employed to derive estimates of intercellular CO2 concentration (Ci) across a range of spatial and temporal scales in a low productivity rain forest ecosystem dominated by the conifer Dacrydium cupressinum Lamb. in New Zealand. Estimates of shoot and canopy Ci across temporal scales ranging from minutes to years were remarkably similar (range of 274-294 micromol mol(-1)). The gradual increase in shoot Ci with depth in the canopy was more likely attributable to decreases in A resulting from lower irradiance (Q) than to increases in g, due to changes in air saturation deficit (D). The lack of marked vertical gradients in A and g(s) at saturating Q through the canopy and the low seasonal variability in environmental conditions contributed to the efficacy of scaling Ci. However, the canopy Ci estimate calculated from the carbon isotope composition of respired ecosystem CO2 (delta13CR; 236 micromol mol(-1)) was much lower than other estimates of canopy Ci. Partitioning delta13CR into four components (soil, roots, litter and foliage) indicated root respiration as the dominant (> 50%) contributor to delta13CR. Variable time lags and differences in isotopic composition during photosynthesis and respiration make the direct estimation of canopy Ci from delta 13CR problematic. PMID- 17080603 TI - Phosphorus nutrition and mycorrhiza effects on grass leaf growth. P status- and size-mediated effects on growth zone kinematics. AB - This study tested whether leaf elongation rate (LER, mm h(-1)) and its components -average relative elemental growth rate (REGRavg, mm mm(-1) h(-1)) and leaf growth zone length (L(LGZ), mm)--are related to phosphorus (P) concentration in the growth zone (P(LGZ) mg P g(-1) tissue water) of Lolium perenne L. cv. Condesa and whether such relationships are modified by the arbuscular mycorrhizal fungus (AMF) Glomus hoi. Mycorrhizal and non-mycorrhizal plants were grown at a range of P supply rates and analysed at either the same plant age or the same tiller size (defined by the length of the sheath of the youngest fully expanded leaf). Both improved P supply (up to 95%) and AMF (up to 21%) strongly increased LER. In tillers of even-aged plants, this was due to increased REGRavg and L(LGZ). In even-sized tillers, it was exclusively due to increased REGRavg. REGRavg was strictly related to P(LGZ) (r2 = 0.95) and independent of tiller size. Conversely, L(LGZ) strictly depended on tiller size (r2 = 0.88) and not on P(LGZ). Hence, P status affected leaf growth directly only through effects on relative tissue expansion rates. Symbiosis with AMF did not modify these relationships. Thus, no evidence for P status-independent effects of AMF on LER was found. PMID- 17080604 TI - Impact of elevated UV-B radiation on photosynthetic electron transport, primary productivity and carbon allocation in estuarine epipelic diatoms. AB - Epipelic diatoms are important components of microphytobenthic biofilms. Cultures of four diatom species (Amphora coffeaeformis, Cylindrotheca closterium, Navicula perminuta and Nitzschia epithemioides) and assemblages of mixed diatom species collected from an estuary were exposed to elevated levels of ultraviolet-B (UV-B) radiation. Short exposures to UV-B resulted in decreases in photosystem II (PSII) photochemistry, photosynthetic electron transport, photosynthetic carbon assimilation and changes in the pattern of allocation of assimilated carbon into soluble colloidal, extracellular polysaccharides (EPS) and glucan pools. The magnitude of the effects of the UV-B treatments varied between species and was also dependent upon the photosynthetically active photon flux density (PPFD) to which the cells were also exposed, with effects being greater at lower light levels. Both increases in nonphotochemical quenching of excitation energy in the pigment antennae and photodamage to the D1 reaction centres contributed to decreases in PSII photochemistry. All species demonstrated a rapid ability to recover from perturbations of PSII photochemistry, with some species recovering during the UV-B exposure period. Some of the perturbations induced in carbon metabolism were independent of effects on PSII photochemistry and photosynthetic electron transport. Elevated UV-B can significantly inhibit photosynthetic performance, and modify carbon metabolism in epipelic diatoms. However, the ecological effects of UV-B at the community level are difficult to predict as large variations occur between species. PMID- 17080605 TI - Adjustments in hydraulic architecture of Pinus palustris maintain similar stomatal conductance in xeric and mesic habitats. AB - We investigated relationships between whole-tree hydraulic architecture and stomatal conductance in Pinus palustris Mill. (longleaf pine) across habitats that differed in soil properties and habitat structure. Trees occupying a xeric habitat (characterized by sandy, well-drained soils, higher nitrogen availability and lower overstory tree density) were shorter in stature and had lower sapwood to-leaf area ratio (A(S):A(L)) than trees in a mesic habitat. The soil-leaf water potential gradient (psiS - psiL) and leaf-specific hydraulic conductance (kL) were similar between sites, as was tissue-specific hydraulic conductivity (Ks) of roots. Leaf and canopy stomatal conductance (gs and Gs, respectively) were also similar between sites, and they tended to be somewhat higher at the xeric site during morning hours when vapour pressure deficit (D) was low. A hydraulic model incorporating tree height, A(S):A(L) and psiS-psiL accurately described the observed variation in individual tree G(Sref) (G(S) at D = 1 kPa) across sites and indicated that tree height was an important determinant of G(Sref) across sites. This, combined with a 42% higher root-to-leaf area ratio (A(R):A(L)) at the xeric site, suggests that xeric site trees are hydraulically well equipped to realize equal--and sometimes higher potential for conductance compared with trees on mesic sites. However, a slightly more sensitive stomatal closure response to increasing D observed in xeric site trees suggests that this potential for higher conductance may only be reached when D is low and when the capacity of the hydraulic system to supply water to foliage is not greatly challenged. PMID- 17080606 TI - Reduced levels of volatile emissions in jasmonate-deficient spr2 tomato mutants favour oviposition by insect herbivores. AB - Tomato plants release volatile organic compounds (VOCs) following insect or mechanical damage. In this study, the constitutive and wound-induced emission levels of VOCs in suppressor of prosystemin-mediated responses2 (spr2) mutant plants, compromised in linolenic acid (LA) and jasmonic acid (JA) synthesis, and in 35S::prosystemin (35S::prosys) plants, having upregulated direct defence responses, were compared. The spr2 mutants produced constitutively lower levels of VOCs, which were nonetheless increased in response to (a)biotic damage, although at lower levels than wild-type (Wt) and 35S::prosys plants. No significant differences in VOC emissions were detected between the latter two genotypes, thereby suggesting that systemin does not regulate indirect defence responses, whereas differences in fatty acid composition in spr2 plants led to the predominant emission of saturated C6 volatiles in response to wounding. The expression of 1-deoxy-D-xylulose 5-phosphate synthase (DXS2), a key gene involved in VOC synthesis in the chloroplast, was only upregulated in Manduca sexta L. damaged Wt and 35S::prosys plants. However, its expression was restored in spr2 plants by exogenous LA or JA, suggesting that abated VOC emissions in spr2 plants are correlated with lowered DXS2 expression. Bioassays with two different insects showed that adult females significantly preferred spr2 plants, indicating that lowered levels of VOCs in tomato influence plant selection by insects during oviposition. PMID- 17080607 TI - Identification of cold acclimation-responsive Rhododendron genes for lipid metabolism, membrane transport and lignin biosynthesis: importance of moderately abundant ESTs in genomic studies. AB - We have previously analysed expressed sequence tags (ESTs) from non-acclimated (NA) and cold-acclimated (CA) Rhododendron leaves, and identified highly abundant complementary DNAs (cDNAs) possibly involved in cold acclimation. A potentially significant, but relatively unexplored, application of these EST data sets is the study of moderately abundant cDNAs, such as those picked only 1-3 times from each Rhododendron EST library containing approximately 430 ESTs. Using statistical tests and Northern blots, we established that the probability of differential expression of moderately abundant cDNAs based on the EST data is, indeed, a reasonably accurate predictor of their 'true' upregulation or downregulation as 11 out of 13 cDNAs (85%) studied fit this criterion. The analyses also revealed four aspects of cold acclimation in Rhododendron leaf tissues. Firstly, the concomitant upregulation of long-chain acyl-coenzyme A (acyl-CoA) synthetase, CTP:cholinephosphate cytidylyltransferase and delta-12 fatty acid desaturase in CA leaf tissues suggests that phospholipid biosynthesis and desaturation are important components of cold hardening in Rhododendron. Secondly, upregulation of plastidic nicotinamide adenine dinucleotide phosphatemalic enzyme (NADP-ME) in CA tissues suggests that malate is an important source of acetyl-CoA used for fatty acid biosynthesis during cold acclimation. Thirdly, down-regulation of plasma membrane intrinsic protein (PIP)2-1 aquaporin and upregulation of gated outward rectifying K+ channel (GORK) in CA tissues may be associated with the protection of overwintering leaves from freeze-induced cellular dehydration. Fourthly, upregulation of coumarate 3-hydroxylase may be associated with cell wall thickening in CA tissues. Physiological implications of these results, which reveal potentially novel regulations of cold acclimation in overwintering woody evergreens, are discussed. This work highlights the importance of also investigating low/moderately abundant ESTs (in addition to highly abundant ones) in genomic studies, in that it offers an effective strategy for identifying stress-related genes, especially when large-scale cDNA sequencing/microarray studies are not possible. PMID- 17080608 TI - Functional coordination between leaf gas exchange and vulnerability to xylem cavitation in temperate forest trees. AB - We examined functional coordination among stem and root vulnerability to xylem cavitation, plant water transport characteristics and leaf traits in 14 co occurring temperate tree species. Relationships were evaluated using both traditional cross-species correlations and phylogenetically independent contrast (PIC) correlations. For stems, the xylem tension at which 50% of hydraulic conductivity was lost (psi50) was positively associated (P < 0.001) with specific conductivity (K(S)) and with mean hydraulically weighted xylem conduit diameter (D(h-w)), but was only marginally (P = 0.06) associated with leaf specific conductivity (K(L)). The PIC correlation for each of these relationships, however, was not statistically significant. There was also no relationship between root psi50 and root K(S) in either cross-species or PIC analysis. Photosynthetic rate (A) and stomatal conductance (g(s)) were strongly and positively correlated with root psi50 in the cross-species analysis (P < 0.001), a relationship that was robust to phylogenetic correction (P < 0.01). A and g(s) were also positively correlated with stem psi50 in the cross-species analysis (P = 0.02 and 0.10, respectively). However, only A was associated with stem psi50 in the PIC analysis (P = 0.04). Although the relationship between vulnerability to cavitation and xylem conductivity traits within specific organs (i.e. stems and roots) was weak, the strong correlation between g(s) and root psi50 across species suggests that there is a trade-off between vulnerability to cavitation and water transport capacity at the whole-plant level. Our results were therefore consistent with the expectation of coordination between vulnerability to xylem cavitation and the regulation of stomatal conductance, and highlight the potential physiological and evolutionary significance of root hydraulic properties in controlling interspecific variation in leaf function. PMID- 17080609 TI - Higher rates of leaf gas exchange are associated with higher leaf hydrodynamic pressure gradients. AB - Steady-state leaf gas-exchange parameters and leaf hydraulic conductance were measured on 10 vascular plant species, grown under high light and well-watered conditions, in order to test for evidence of a departure from hydraulic homeostasis within leaves as hydraulic conductance varied across species. The plants ranged from herbaceous crop plants to mature forest trees. Across species, under standardized environmental conditions (saturating light, well watered), mean steady-state stomatal conductance to water vapour (g(w)) was highly correlated with mean rate of CO2 assimilation (A) and mean leaf hydraulic conductance normalized to leaf area (k(leaf)). The relationship between A and g(w) was well described by a power function, while that between A and k(leaf) was highly linear. Non-linearity in the relationship between g(w) and k(leaf) contributed to an increase in the hydrodynamic (transpiration-induced) water potential drawdown across the leaf (delta psi(leaf)) as k(leaf) increased across species, although across the 10 species the total increase in delta psi(leaf) was slightly more than twofold for an almost 30-fold increase in g(w). Higher rates of leaf gas exchange were therefore associated with higher k(leaf) and higher leaf hydrodynamic pressure gradients. A mechanistic model incorporating the stomatal hydromechanical feedback loop is used to predict the relationship between delta psi(leaf) and k(leaf), and to explore the coordination of stomatal and leaf hydraulic properties in supporting higher rates of leaf gas exchange. PMID- 17080610 TI - Molecular identification, heterologous expression and properties of light insensitive plant catalases. AB - Most catalases are inactivated by light in a heme-sensitized and O2-dependent reaction. In leaves of the alpine plant Homogyne alpina and in the peroxisomal cores of Helianthus annuus, light-insensitive catalases were observed. For the catalases Hacat1 of H. alpina and HnncatA3 of H. annuus, cDNA clones were obtained. Expression of recombinant active enzymes in insect cells confirmed that they coded for light-insensitive catalases. Kinetic and catalytic properties of light-sensitive or light-insensitive catalases did not differ substantially. However, the specific activity of the latter was markedly lower. The light insensitive catalase HaCAT-1 was not resistant against inactivation by superoxide. Amino acid sequences of the light-insensitive catalases HaCAT-1 and HNNCATA3 were highly identical. They showed only a few exceptional amino acid substitutions at positions that are highly conserved in other catalases. These appeared to be localized mainly in a surface cavity at the entrance of a minor channel leading to the central heme, suggesting that this region played some, though yet undefined, role for light sensitivity. While the replacement of a highly conserved His by Thr225 was the most unique substitution, a single exchange of His225 by Thr in the light-sensitive catalase SaCAT-1 by mutagenesis was not sufficient to reduce its sensitivity to photoinactivation. PMID- 17080611 TI - Acyl-homoserine lactones modulate the settlement rate of zoospores of the marine alga Ulva intestinalis via a novel chemokinetic mechanism. AB - Bacteria utilize quorum sensing to regulate the expression of cell density dependant phenotypes such as biofilm formation and virulence. Zoospores of the marine alga Ulva intestinalis exploit the acyl-homoserine lactone (AHL) quorum sensing system to identify bacterial biofilms for preferential settlement. Here, we demonstrate that AHLs act as strong chemoattractants for Ulva zoospores. Chemoattraction does not involve a chemotactic orientation towards the AHL source. Instead, it occurs through a chemokinesis in which zoospore swimming speed is rapidly decreased in the presence of AHLs. The chemoresponse to AHLs was dependant on the nature of the acyl side chain, with N-(3-oxododecanoyl) homoserine lactone (30-C12-HSL) being the most effective signal molecule. Mean zoospore swimming speed decreased more rapidly over wild-type biofilms of the marine bacteria Vibrio anguillarum relative to biofilms of the vanM mutant, in which AHL synthesis is disrupted. These data implicate a role for AHL-mediated chemokinesis in the location and preferential settlement of Ulva zoospores on marine bacterial assemblages. Exposure to AHLs did not inhibit the negative phototaxis of Ulva zoospores, indicating that chemoattraction to bacterial biofilms does not preclude the response to a light stimulus in substrate location. PMID- 17080612 TI - gid1, a gibberellin-insensitive dwarf mutant, shows altered regulation of probenazole-inducible protein (PBZ1) in response to cold stress and pathogen attack. AB - A recessive gibberellin (GA)-insensitive dwarf mutant of rice, gibberellin insensitive dwarf1 (gid1), has been identified, which shows a severe dwarf phenotype and contains high concentrations of endogenous GA. To elucidate the function of gid1, proteins regulated downstream of gid1 were analysed using a proteomic approach. Proteins extracted from suspension-cultured cells of gid1 and its wild type were separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Of a total of 962 proteins identified from the suspension-cultured cells, 16 were increased and 14 were decreased in gid1 compared with its wild type. Among the proteins hyper-accumulated in gid1 were osmotin, triosephosphate isomerase, probenazole inducible protein (PBZ1) and pathogenesis-related protein 10. Of these four genes, only the expression of PBZ1 was increased by exogenous GA3 application. Expression of this gene was also enhanced in shoots of the wild type by cold stress or by rice blast fungus infection. Under normal growth conditions, there was more PBZ1 protein in gid1 than in the wild type. In addition, gid1 showed increased tolerance to cold stress and resistance to blast fungus infection. The entcopalyl diphosphate synthase (OsCPS) genes, which encode enzymes at the branch point between GA and phytoalexin biosynthesis, were expressed differentially in gid1 relative to the wild type. Specifically, OsCPS1, which encodes an enzyme in the GA biosynthesis pathway, was down-regulated and OsCPS2 and OsCPS4, which encode enzymes in phytoalexin biosynthesis, were up-regulated in gid1. These results suggest that the expression of PBZ1 is regulated by GA signalling and stress stimuli, and that gid1 is involved in tolerance to cold stress and resistance to blast fungus. PMID- 17080613 TI - Stem and leaf gas exchange and their responses to fire in a north Australian tropical savanna. AB - We measured stem CO2 efflux and leaf gas exchange in a tropical savanna ecosystem in northern Australia, and assessed the impact of fire on these processes. Gas exchange of mature leaves that flushed after a fire showed only slight differences from that of mature leaves on unburned trees. Expanding leaves typically showed net losses of CO2 to the atmosphere in both burned and unburned trees, even under saturating irradiance. Fire caused stem CO2 efflux to decline in overstory trees, when measured 8 weeks post-fire. This decline was thought to have resulted from reduced availability of C substrate for respiration, due to reduced canopy photosynthesis caused by leaf scorching, and to priority allocation of fixed C towards reconstruction of a new canopy. At the ecosystem scale, we estimated the annual above-ground woody-tissue CO2 efflux to be 275 g C m(-2) ground area year(-1) in a non-fire year, or approximately 13% of the annual gross primary production. We contrasted the canopy physiology of two co-dominant overstory tree species, one of which has a smooth bark on its branches capable of photosynthetic re-fixation (Eucalyptus miniata), and the other of which has a thick, rough bark incapable of re-fixation (Eucalyptus tetrodonta). Eucalyptus miniata supported a larger branch sapwood cross-sectional area in the crown per unit subtending leaf area, and had higher leaf stomatal conductance and photosynthesis than E. tetrodonta. Re-fixation by photosynthetic bark reduces the C cost of delivering water to evaporative sites in leaves, because it reduces the net C cost of constructing and maintaining sapwood. We suggest that re-fixation allowed leaves of E. miniata to photosynthesize at higher rates than those of E. tetrodonta, while the two invested similar amounts of C in the maintenance of branch sapwood. PMID- 17080614 TI - Growth and ethylene evolution by shade and sun ecotypes of Stellaria longipes in response to varied light quality and irradiance. AB - Plants growing in the shade receive both low light irradiance and light enriched in far red (FR) (i.e., light with a low red (R) to FR ratio). In an attempt to uncouple the R/FR ratio effects from light irradiance effects, we utilized Stellaria longipes because this species has two distinct natural population ecotypes, alpine (dwarf) and prairie (tall). The alpine population occupies the open, sun habitat. By contrast, the prairie population grows in the shade of other plants. Both 'sun' and 'shade' ecotypes responded with increased stem elongation responses under low irradiance, relative to growth under 'normal' irradiance, and this increased growth was proportionally similar. However, only the shade ecotype had increased shoot elongation in response to a low R/FR ratio. By contrast, the sun ecotype showed increased stem elongation in response to increasing R/FR ratio. Varying the R/FR ratios had no significant effect on ethylene evolution in either sun or shade ecotype. Under low irradiance, only the sun ecotype showed a significantly changed (decreased) ethylene evolution. We conclude that R/FR ratio and irradiance both regulate growth, and that irradiance can also influence ethylene evolution of the sun ecotype. By contrast, R/FR ratio and irradiance, while having profound influences on growth of the shade ecotype, do not appear to regulate these growth changes via effects on ethylene production. PMID- 17080615 TI - Genotypic difference in canopy diffusive conductance measured by a new remote sensing method and its association with the difference in rice yield potential. AB - There have been few practical ways of measuring physiological determinants of rice yield. Rapid evaluation of yield determination traits may expedite breeding of high-yielding rice. Here, we report a new remote-sensing technique for the evaluation of canopy ecophysiological status under field conditions developed based on simultaneous measurements of sunlit and suddenly shaded canopy temperatures. This technique has the advantage of instantaneous estimation of aerodynamic resistance (r(a)) and canopy diffusive resistance (r(c) without measuring wind velocity. Canopy diffusive conductance (1 / r(c)) estimated by the remote sensing method was closely related to leaf stomatal conductance (g(s)) measured with a portable gas exchange system. This result supported the validity of this new method for quantitative estimation of canopy physiological characteristics. Significant genotypic differences were obtained in canopy-air temperature difference (Tc-Ta), r(c) and 1 / r(c) during the 2-week period preceding full heading for two years, and 1 / r(c) was highly correlated with crop growth rate (CGR), which was closely related to the final yield. These results suggest that 1 / r(c) can be an effective criterion for the selection of high-yielding rice genotypes, and the remote sensing technique proposed here can be a powerful tool for the rapid evaluation of 1 / r(c) under field conditions. PMID- 17080616 TI - Ca2+ transient induced by extracellular changes in osmotic pressure in Arabidopsis leaves: differential involvement of cell wall-plasma membrane adhesion. AB - We investigated the mechanism underlying the perception of extracellular changes in osmotic pressure in Vallisneria gigantea Graebner and transgenic Arabidopsis thaliana (L.) Heynh. expressing cytoplasmic aequorin. Hypertonic and hypotonic treatments of A. thaliana leaves each rapidly induced a Ca2+ transient. Both responses were essentially dependent on the presence of extracellular Ca2+ and were sensitive to Gd3+ a potential blocker of stretch-activated Ca2+ channels. Immediately after plasmolysis caused by hypertonic treatment and subsequent deplasmolysis caused by hypotonic treatment, the cells did not respond to a second hypertonic treatment and exhibited an impaired adhesion of the plasma membrane (PM) to the cell wall (CW). Recovery of the responsiveness required about 6 h. By contrast, no refractory phenomenon was observed in response to hypotonic treatment. Pretreatment with cellulase completely inhibited the Ca2+ transient induced by hypertonic treatment, but it did not affect the response to hypotonic treatment. V. gigantea mesophyll cells pretreated with cellulase exhibited an impaired adhesion of the PM to the CW. The leaf cells of multicellular plants can respond to both hypertonic and hypotonic treatments through the stretch-activated Ca2+ channels, whereas cellulase-sensitive adhesion of the PM to the CW is involved only in the response to hypertonic treatment. PMID- 17080617 TI - Evidence for altered polar and lateral auxin transport in the gravity persistent signal (gps) mutants of Arabidopsis. AB - Plant shoots do not respond when they are reoriented relative to gravity at 4 degrees C. However, when returned to vertical at room temperature, these organs bend in response to the previous cold gravistimulation. The inflorescence stem of the Arabidopsis thaliana gravity persistent signal (gps) mutants respond abnormally after the cold gravistimulation: gps1 does not bend when returned to room temperature, gps2 bends the wrong way and gps3 over-responds, curving past the predicted angle. In wild type and the mutants, basipetal auxin transport in the inflorescence stem was abolished at 4 degrees C but restored when plants were returned to room temperature. In gps1, auxin transport was increased; in both gps2 and gps3, no significant difference was found when compared to wild type. Expression of the auxin-inducible P(IAA2)::GUS reporter gene, indicated that auxin-induced gene expression was redistributed to the lower side of the inflorescence stem in wild type after gravistimulation at 4 degrees C. In gps1, no asymmetries in P(IAA2)::GUS expression were seen. In gps2, P(IAA2)::GUS expression was localized to the upper side of the stem and in gps3, asymmetric P(IAA2):GUS expression was extended throughout the elongation zone of the inflorescence stem. These results are consistent with altered lateral Indole-3 acetic-acid (IAA) gradients being responsible for the phenotype of each mutant. PMID- 17080618 TI - Effects of growth light and nitrogen nutrition on the organization of the photosynthetic apparatus in leaves of a C4 plant, Amaranthus cruentus. AB - Properties of C4 photosynthesis were examined in Amaranthus cruentus L. (NAD malic enzyme (ME) subtype, dicot) grown under different light and nitrogen (N) conditions, from the viewpoint of N investment into their photosynthetic components. In low-light (LL) leaves, chlorophyll content per leaf area was greater and chlorophyll alb ratio was lower than in high-light (HL) leaves. These indicate that LL leaves invest more N into their light-harvesting systems. However, this N investment did not contribute to the increase in the quantum yield of photosynthesis on the incident photon flux density (PFD) basis (Qi) in LL leaves. N allocation to ribulose 1,5-bisphosphate carboxylasel oxygenase (Rubisco) was significantly higher in HL-high N (HN) leaves than in other leaves. On the other hand, N allocation to C4 enzymes [phosphoenolpyruvate carboxylase (PEPC) and pyruvate Pi dikinase (PPDK)] was unaffected by the growth conditions. Maximum photosynthetic rates (Pmax) per Rubisco content were similar irrespective of the growth light treatments. Carbon isotope ratios (delta13 C) in the leaf dry matter were more negative in LL leaves than in HL leaves (LL = -19.3% per hundred, HL = -16.0% per hundred) and independent of leaf N. Vein density was highest in HL-HN leaves, and leaf thickness was unaffected by the growth light treatments. From these results, we conclude that A. cruentus leaves would not acclimate efficiently to low growth light. PMID- 17080619 TI - Tomato seed germination: regulation of different response modes by phytochrome B2 and phytochrome A. AB - Lycopersicon esculentum seeds germinate after rehydration in complete darkness. This response was inhibited by a far-red light (FR) pulse, and the inhibition was reversed by a red light (R) pulse. Comparison of germination in phytochrome deficient mutants (phyA, phyB1, phyB2, phyAB1, phyB1B2 and phyAB1B2) showed that phytochrome B2 (PhyB2) mediates both responses. The germination was inhibited by strong continuous R (38 micromol m(-2) s(-1)), whereas weak R (28 nmol m(-2) s( 1)) stimulated seed germination. Hourly applied R pulses of the same photon fluence partially replaced the effect of strong continuous R. This response was called 'antagonistic' because it counteracts the low fluence response (LFR) induced by a single R pulse. This antagonistic response might be an adaptation to a situation where the seeds sit on the soil surface in full sunlight (adverse for germination), while weak R might reflect that situation under a layer of soil. Unexpectedly, the effects of continuous R or repeated R pulses were mediated by phytochrome A (PhyA). We therefore suggest that low levels of PhyA in its FR absorbing form (Pfr) cause inhibition of seed germination produced either by extended R irradiation (by degradation of PhyA-Pfr) or by extended FR irradiation [keeping a low Pfr/R-absorbing form (Pr) ratio]. PMID- 17080620 TI - Responses of spinach leaf mitochondria to low N availability. AB - Low N availability induces carbohydrate accumulation in leaf cells, which often causes suppression of photosynthesis. Under low N supply, excess carbohydrates would be preferentially respired by the non-phosphorylating pathways, such as the alternative oxidase (AOX) and uncoupling protein (UCP), which would suppress the excessive increase in the ratio of C to N (C/N ratio). In leaves, however, responses of these pathways to the low N stress are still unknown. We examined the mitochondrial respiratory pathways in spinach leaves grown at three different N availabilities to clarify whether the respiratory pathways change depending on the N availabilities. With the decrease in N availability, leaf respiratory rates per leaf area decreased, but the rates on the leaf N basis were comparable. Using fumarase activities of whole leaf extracts and isolated mitochondria, we estimated mitochondrial protein contents per leaf N. The contents increased with the decrease in the N availability, that is, at the low N availability, N was preferentially invested into mitochondria. On the mitochondrial protein basis, capacities of cytochrome pathway (CP) and cytochrome c oxidase (COX) were comparable regardless of the N availabilities, whereas both AOX capacity and the amounts of AOX protein increased with the decrease in the N availability. Some enzymes of tricarboxylic acid (TCA) cycle, especially NAD-dependent malic enzyme (NAD-ME), showed higher capacities under lower N. On the other hand, amounts of UCP did not differ amongst the N availabilities. These results indicated that, under low N stress, AOX will be preferentially up-regulated and will efficiently consume excess carbohydrates, which leads to suppressing the rise in the C/N ratio to a moderate level. PMID- 17080621 TI - Low growth temperatures modify the efficiency of light use by photosystem II for CO2 assimilation in leaves of two chilling-tolerant C4 species, Cyperus longus L. and Miscanthus x giganteus. AB - Two C4 plants, Miscanthus x giganteus and Cyperus longus L., were grown at suboptimal growth temperatures and the relationships between the quantum efficiencies of photosynthetic electron transport through photosystem II (PSII) (PSII operating efficiency; Fq'/Fm') and CO2 assimilation (phiCO2) in leaves were examined. When M. x giganteus was grown at 10 degrees C, the ratio of the PSII operating efficiency to phiCO2 increased relative to that found in leaves grown at 14 and 25 degrees C. Similar increases in the Fq'/Fm': phiCO2 occurred in the leaves of two C. longus ecotypes when the plants were grown at 17 degrees C, compared to 25 degrees C. These elevations of Fq'/Fm': phiCO2 at low growth temperatures were not attributable to the development of anthocyanins, as has been suggested for maize, and were indicative of the operation of an alternative sink to CO2 assimilation for photosynthetic reducing equivalents, possibly oxygen reduction via a Mehler reaction, which would act as a mechanism for protection of PSII from photoinactivation and damage. Furthermore, in M. x giganteus grown at 10 degrees C, further protection of PSII was effected by a 20-fold increase in zeaxanthin content in dark-adapted leaves, which was associated with much higher levels of non-photochemical quenching of excitation energy, compared to that observed in leaves grown at 14 and 25 degrees C. These differences may explain the long growing season and remarkable productivity of this C4 plant in cool climates, even in comparison to other C4 species such as C. longus, which occur naturally in such climates. PMID- 17080622 TI - [Nurses as prescribers]. PMID- 17080623 TI - [Testing an instrument measuring Italian nurses' knowledge and attitudes regarding pain]. AB - AIM: To test the construct validity, test-retest reliability, and internal consistency of the Nurses' Knowledge and Attitudes Survey Regarding Pain translated from English to Italian. DESIGN: Validation study. SETTING: Oncology, long term care wards and hospice of 5 different italian regions. SAMPLE: 157 nurses divided in four groups according to the setting: students (29), general medical wards (35), oncology (43) and hospice (50). METHODS: The instrument was translated and adapted from English to Italian by a group of experts in pain and by pilot-testing. It was completed by 90% of the sample and the percentage of correct answers and alpha Cronbach were calculated. The test-retest reliability was measured having the questionnaire completed after a 14 days time interval, by the nurses of medical wards. RESULTS: The instrument discriminated between levels of expertise. Overall, the average percentage of correct answers was 52.7%: 40.7% for student nurses, 46% for general medicine nurses, 56.3% for oncology nurses and 62% for hospice nurses (P<0.001). Test-retest reliability was r = 0.97 and Cronbach's alfa was 0.69. CONCLUSION: These results contribute to the validity and reliability of the Italian version of the Nurses' Knowledge and Attitudes Survey Regarding Pain, show a limited knowledge in the four groups of nurses and provide the basis for other studies in oncology and palliative settings. PMID- 17080624 TI - [Nurses and doctors in the management of arrhytmic complications: the case of electric storm in patients with an implantable cardioverter defibrillator]. AB - The evolution of arrhythmologic technology introduced new therapeutic opportunities for cardiology patients. Competence, continuity of care, integration and collaboration between medical doctors and nurses allow a safe management of short and long term complications. A typical example of a new arrhythmic complication is the "electrical storm" or "arrhythmic warm up" in patients implanted with a cardioverter-defibrillator (ICD). The definition and epidemiology of the electrical storm stress the relevance of this clinical situation; its dramatic emergency involves several aspects of nursing care, clinical, prognostic, ethical and deontological. The definition and implementation of an integrated protocol for the treatment of this condition is described and ethical and unresolved questions are rised. One of these problems is the recommendation to offer the patient the opportunity to receive information about the option of inactivating the ICD. PMID- 17080625 TI - [Knowledge and lifestyles of patients: a survey on patients admitted with acute coronary syndrome in the CCU of an Italian hospital]. AB - AIM AND METHODS: The lifestyles before hospital admission, knowledge on their illness and lifestyles after the acute coronary event were analysed with questionnaires, in three different samples of patients: a. all the patients admitted for acute coronary event from may 2003 to may 2005 to explore lifestyles before acute coronary event (416 patients) b. all the patients admitted from may 2003 to april 2004 (before the start of health information meetings organised by nurses), to explore the knowledge of the illness and its causes (132 patients) c. a sample of 83 patients followed in day hospital, to explore the lifestyles after the acute coronary event. RESULTS: Lifestyles before the event. Most patients have incorrect lifestyles: 50% eat cheese every day and never exercise for at least 30 minutes everyday. Even after the acute coronary event, some incorrect lifestyles are still present. Seventy-five percent of patients have incorrect or insufficient knowledge on illness and risk factors at discharge and only 50% is willing to increase the amount of fruit and vegetables in their diet. CONCLUSIONS: Although confirmed by other studies, these results are worrying and call for the systematic adoption of secondary prevention strategies with effective interventions aimed at increasing knowledge and modifying lifestyles. PMID- 17080626 TI - [An health education program for patients admitted to CCU for an acute coronary event]. AB - In spite of the broad recognition of the importance of health education, time for structured one-to -one initiatives of health education during the hospital stay is limited. The organization of an health education meeting for patients admitted to CCU for an acute coronary event is described. METHODS: The planning and implementation of the initiative lasted two years and involved 7 nurses and one doctor. The organization required efforts related to the event itself (preparation of training aids, identification or contents and methods for delivery) but also organizative changes. Dietitians in fact had to be involved because the healthy diet recommended was different from the hospital diet. The assessment of the effectiveness of the health education was also planned: administration of a questionnaire to explore lifestyles and knowledge of the illness before and after the meeting; phone interviews after 3, 6 and 12 months from the meeting. RESULTS: Since may 2003, in the first 3 years 74 meetings have been organised, involving 507 patients and 329 relatives. Each meeting lasts 2 hours and contents delivered encompass the coronary event, risk factors and their modification, healthy lifestyles. Initial preliminary results on the impact of the meeting on lifestyle changes are promising. Initiatives are ongoing to include this activity among officially recognised nursing activities. PMID- 17080627 TI - [The OSCE method (Objective Structured Clinical Examination) in the university course for nurses in Udine]. AB - The problems and discrepancies occurring in the final evaluation of nursing students brought about the introduction of the OSCE method (Objective Structured Clinical Examination) in the University Course for nurses. AIM: of the article is to describe our experience and first results. METHODS: Sixty-seven Ist year students were assessed with the OSCE and asked to perform 3 tests to assess theoretical knowledge; practical skills and diagnostic reasoning. RESULTS: 82% passed the three tests. The best results were achieved for the diagnositc reasoning (87%) and the higher number of failures in the practical skills test (57%). The students who had previously passed the exams of the nursing courses had a lower number of failures (3% vs 29%). CONCLUSIONS: exposure to the skills assessed and to the differences between what is thaugh in the classroom and what the students observe in practice. A close collaboration is needed with ward nurses to reduce the gap between theory and practice and the variability of assessment criteria. PMID- 17080628 TI - [Prescriptions by non medical personnel]. PMID- 17080629 TI - [What is the cost of health in low income countries]. PMID- 17080630 TI - [Children as "collateral damages" in Lebanon]. PMID- 17080631 TI - Evidence from Amazonian forests is consistent with isohydric control of leaf water potential. AB - Climate modelling studies predict that the rain forests of the Eastern Amazon basin are likely to experience reductions in rainfall of up to 50% over the next 50-100 years. Efforts to predict the effects of changing climate, especially drought stress, on forest gas exchange are currently limited by uncertainty about the mechanism that controls stomatal closure in response to low soil moisture. At a through-fall exclusion experiment in Eastern Amazonia where water was experimentally excluded from the soil, we tested the hypothesis that plants are isohydric, that is, when water is scarce, the stomata act to prevent leaf water potential from dropping below a critical threshold level. We made diurnal measurements of leaf water potential (psi 1), stomatal conductance (g(s)), sap flow and stem water potential (psi stem) in the wet and dry seasons. We compared the data with the predictions of the soil-plant-atmosphere (SPA) model, which embeds the isohydric hypothesis within its stomatal conductance algorithm. The model inputs for meteorology, leaf area index (LAI), soil water potential and soil-to-leaf hydraulic resistance (R) were altered between seasons in accordance with measured values. No optimization parameters were used to adjust the model. This 'mechanistic' model of stomatal function was able to explain the individual tree-level seasonal changes in water relations (r2 = 0.85, 0.90 and 0.58 for psi 1, sap flow and g(s), respectively). The model indicated that the measured increase in R was the dominant cause of restricted water use during the dry season, resulting in a modelled restriction of sap flow four times greater than that caused by reduced soil water potential. Higher resistance during the dry season resulted from an increase in below-ground resistance (including root and soil-to-root resistance) to water flow. PMID- 17080632 TI - Effects of red, far-red and blue light in maintaining growth in latitudinal populations of Norway spruce (Picea abies). AB - Seedlings of trees with a free growth pattern cease growth when night-lengths become shorter than a critical value, and this critical night-length (CNL) decreases with increasing latitude of origin. In northern populations, the light quality also appears to play an important role and a clinal variation in requirement for far-red (FR) light has been documented. In this study we dissected the light quality requirements for maintaining growth in different latitudinal populations of Norway spruce (Picea abies (L.) H. Karst.) using light emitting diodes for red (R), FR and blue (B) light, as 12 h day extension to provide 24 h photoperiod. At equal spectral photon flux, FR light was more effective than R light in maintaining growth, and the requirement of both R and FR increased with northern latitude of origin. One-to-one mixtures of R and FR light were more effective in maintaining growth than either FR or R light alone, indicating a possible interaction between R and FR light maintaining growth. Using the blue light as day extension could not prevent growth cessation in any of the populations, but delayed the bud set slightly in all populations. Our results suggest that phytochrome(s) are the primary photoreceptors in high irradiance responses maintaining growth in Norway spruce seedlings. PMID- 17080633 TI - Characterization of salt tolerance in ectoine-transformed tobacco plants (Nicotiana tabaccum): photosynthesis, osmotic adjustment, and nitrogen partitioning. AB - Ectoine (1,4,5,6-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid) biosynthetic genes (ect. ABC) from Halomonas elongata were introduced to tobacco plants using an Agrobacterium-mediated gene delivery system. The genes for ectoine biosynthesis were integrated in a stable manner into the tobacco genome and the corresponding transcripts were expressed. The concentration of ectoine under salt stress conditions was higher in the roots than in leaves. A close relationship was found between stomatal conductance and the amount of transported nitrogen, suggesting that water transport through the xylem in the stem and transpiration may be involved in nitrogen transport to leaves. The data indicate that the turgor values of the ectoine transgenic lines increased with increasing salt concentration. The data revealed two ways in which ectoine enhanced salinity tolerance of tobacco plants. First, ectoine improved the maintenance of root function so that water is taken up consistently and supplied to shoots under saline conditions. Second, ectoine enhanced the nitrogen supply to leaves by increasing transpiration and by protecting Rubisco proteins from deleterious effects of salt, thereby improving the rate of photosynthesis. PMID- 17080634 TI - Elicitor-induced down-regulation of cell cycle-related genes in tobacco cells. AB - The fungal elicitors, a xylanase from Trichoderma viride and an extract from the cell wall of Phytophthora infestans, are shown to cause a rapid reduction of the mRNA levels of various cell cycle-related genes, including MAP kinase genes and cyclin genes, in cultured tobacco cells (Nicotiana tabacum cv. Xanthi, line XD6S). Pharmacological analyses suggest that the elicitor-induced decrease in Bi type cyclin (Nicta;CycB1;3) and A1-type cyclin (Nicta;CycA1;1) mRNAs may be due to transcriptional repression, and that in D3-type cyclin (Nicta;CycD3;2) mRNA due to destabilization of the mRNA molecule itself. The activity of protein kinases is required for both the activation of defence genes and the repression of cyclin genes. The transcriptional activity of the promoter of the B1-class cyclin gene decreases upon elicitor treatment. The transactivation activity of NtmybA2, a tobacco Myb transcription activator for the M phase-specific cis acting elements in the promoter of the B-type cyclin gene, is inhibited by elicitor treatment. In addition, the mRNA levels of NtmybA2 and two other related genes, NtmybA1 and NtmybB, decrease in response to the elicitor. Finally, we discuss a negative cross-talk between signal transduction pathways for growth and defence responses, which might be important for adaptation to environmental stress by potential pathogens. PMID- 17080635 TI - Internal conductance does not scale with photosynthetic capacity: implications for carbon isotope discrimination and the economics of water and nitrogen use in photosynthesis. AB - Central paradigms of ecophysiology are that there are recognizable and even explicit and predictable patterns among species, genera, and life forms in the economics of water and nitrogen use in photosynthesis and in carbon isotope discrimination (delta). However most previous examinations have implicitly assumed an infinite internal conductance (gi) and/or that internal conductance scales with the biochemical capacity for photosynthesis. Examination of published data for 54 species and a detailed examination for three well-characterized species--Eucalyptus globulus, Pseudotsuga menziesii and Phaseolus vulgaris--show these assumptions to be incorrect. The reduction in concentration of CO2 between the substomatal cavity (Ci) and the site of carbon fixation (Cc) varies greatly among species. Photosynthesis does not scale perfectly with gi and there is a general trend for plants with low gi to have a larger draw-down from Ci to Cc, further confounding efforts to scale photosynthesis and other attributes with gi. Variation in the gi-photosynthesis relationship contributes to variation in photosynthetic 'use' efficiency of N (PNUE) and water (WUE). Delta is an information-rich signal, but for many species only about two-thirds of this information relates to A/gs with the remaining one-third related to A/gi. Using data for three well-studied species we demonstrate that at common WUE, delta may vary by up to 3 per thousand. This is as large or larger than is commonly reported in many interspecific comparisons of delta, and adds to previous warnings about simplistic interpretations of WUE based on delta. A priority for future research should be elucidation of relationships between gi and gs and how these vary in response to environmental conditions (e.g. soil water, leaf-to-air vapour pressure deficit, temperature) and among species. PMID- 17080636 TI - Diurnal variation of the delta 13C of pine needle respired CO2 evolved in darkness. AB - The delta 13C of pine needle CO2 evolved in darkness (delta 13Cr) for slash pine trees (Pinus elliottii) was determined by placing recently collected pine needles in darkness and collecting respired CO2 over a short time period (<15 min). Delta 13Cr measurements were made over several 24 h periods to test the hypothesis that significant variation in delta 13Cr would be observed during a diurnal cycle. The delta 13Cr measurements from the 24 h time series trials showed a consistent midday 13C-enrichment (5-10 per thousand) relative to bulk biomass. The delta 13Cr values became more 13C-depleted at night and following shading, and approached bulk-biomass delta 13C values by dawn. The effect of night-time respired 13C-enriched CO2 on the delta 13C value of the remaining assimilate is shown to be minimal (13C depleted by 0.22 per thousand) under field conditions for P. elliottii needles. PMID- 17080638 TI - Hydraulic efficiency and safety of branch xylem increases with height in Sequoia sempervirens (D. Don) crowns. AB - The hydraulic limitation hypothesis of Ryan & Yoder (1997, Bioscience 47, 235 242) suggests that water supply to leaves becomes increasingly difficult with increasing tree height. Within the bounds of this hypothesis, we conjectured that the vertical hydrostatic gradient which gravity generates on the water column in tall trees would cause a progressive increase in xylem 'safety' (increased resistance to embolism and implosion) and a concomitant decrease in xylem 'efficiency' (decreased hydraulic conductivity). We based this idea on the historically recognized concept of a safety-efficiency trade-off in xylem function, and tested it by measuring xylem conductivity and vulnerability to embolism of Sequoia sempervirens branches collected at a range of heights. Measurements of resistance of branch xylem to embolism did indeed show an increase in 'safety' with height. However, the expected decrease in xylem 'efficiency' was not observed. Instead, sapwood-specific hydraulic conductivities (Ks) of branches increased slightly, while leaf-specific hydraulic conductivities increased dramatically, with height. The latter could be largely explained by strong vertical gradients in specific leaf area. The increase in Ks with height corresponded to a decrease in xylem wall fraction (a measure of wall thickness), an increase in percentage of earlywood and slight increases in conduit diameter. These changes are probably adaptive responses to the increased transport requirements of leaves growing in the upper canopy where evaporative demand is greater. The lack of a safety-efficiency tradeoff may be explained by opposing height trends in the pit aperture and conduit diameter of tracheids and the major and semi-independent roles these play in determining xylem safety and efficiency, respectively. PMID- 17080637 TI - Heat sensitivity of photosynthetic electron transport varies during the day due to changes in sugars and osmotic potential. AB - In water-stressed leaves, accumulation of neutral osmotica enhances the heat tolerance of photosynthetic electron transport. There are large diurnal and day to-day changes in leaf sugar content because of variations in net photosynthetic production, respiration and retranslocation. To test the hypothesis that diurnal and day-to-day variations in leaf sugar content and osmotic potential significantly modify the responses to temperature of photosynthetic electron transport rate, we studied chlorophyll fluorescence rise temperatures (i.e. critical temperatures at break-points in fluorescence versus temperature response curves, corresponding to enhanced damage of PSII centers and detachment of pigment-binding complexes) in the dark at a background of weak far-red light (T(FR)) and under actinic light (T(L)), and responses of foliar photosynthetic electron transport rate to temperature using gas-exchange and chlorophyll fluorescence techniques in the temperate tree Populus tremula L. Sucrose and sorbitol feeding experiments demonstrated strong increases of fluorescence rise temperatures T(FR) and T(L) with decreasing leaf osmotic potential and increasing internal sugar concentration. Similar T(FR) and T(L) changes were observed in response to natural variation in leaf sugar concentration throughout the day. Increases in leaf sugar concentration led to an overall down-regulation of the rate of photosynthetic electron transport (J), but increases in the optimum temperature (Topt) of J. For the entire dataset, Topt varied from 33.8 degrees C to 43 degrees C due to natural variation in sugars and from 33.8 degrees C to 52.6 degrees C in the sugar feeding experiments, underscoring the importance of sugars in modifying the response of J to temperature. However, the correlations between the sugar concentration and fluorescence rise temperature varied between the days. This variation in fluorescence rise temperature was best explained by the average temperature of the preceding 5 or 6 days. In addition, there was a significant year-to-year variation in heat sensitivity of photosynthetic electron transport that was associated with year-to-year differences in endogenous sugar content. Our data demonstrate a diurnal variation in leaf heat tolerance due to changes in sugar concentration, but they also show that this short-term modification in heat tolerance is super-imposed by long-term changes in heat resistance driven by average temperature of preceding days. PMID- 17080639 TI - Comparative studies on the effect of a protein-synthesis inhibitor on aluminium induced secretion of organic acids from Fagopyrum esculentum Moench and Cassia tora L. roots. AB - Aluminium (Al)-induced secretion of organic acids from plant roots is considered a mechanism of Al resistance, but the processes leading to the secretion of organic acids are still unknown. In the present study, a protein-synthesis inhibitor, cycloheximide (CHM), was used to investigate its effect on Al-induced organic acid secretion in a pattern I (rapid exudation of organic acids under Al stress) plant buckwheat (Fagopyrum esculentum Moench) and a pattern II (exudation of organic acids was delayed by several hours under Al stress) plant Cassia tora L. A dose-response experiment showed that the secretion of oxalate by buckwheat roots was not affected by CHM when added in the range from 0 to 50 microM, with or without exposure to 100 microm Al, but the secretion of citrate was completely inhibited by 30 microM CHM in C. tora. A time-course experiment showed that even prolonged exposure to 20 microM CHM did not affect oxalate secretion in buckwheat, but significantly inhibited citrate secretion in C. tora. However, citrate synthase (CS) activity in C. tora was not affected during 12 h exposure to 100 microM Al when compared with that in control roots, although CHM can inhibit CS activity effectively. These results indicated that CS activity was not related to Al-regulated citrate efflux in C. tora. The total protein was decreased by 14.0% and 32.3% in C. tora and buckwheat root tip, respectively, after 3-h treatment with 20 microM CHM. A 3-h pulse with 20 microM CHM completely inhibited citrate efflux in C. tora during the next 6-h exposure to Al, although a small amount of citrate was exuded after 9-h exposure. However, oxalate efflux in buckwheat was not influenced by a similar treatment. In buckwheat, a 3-h pulse with 100 microM Al maintained oxalate secretion at a high level during the next 9 h, with or without CHM treatment. Conversely, in C. tora a 6-h pulse with 100 microM Al induced significant secretion of citrate which was inhibited by the CHM. Taken together, these findings suggest that both de novo synthesis and activation of an anion channel are needed for Al-induced secretion of citrate in C. tora, but in buckwheat the plasma membrane protein responsible for oxalate secretion pre-exists. PMID- 17080640 TI - Ecological physiology of Pereskia guamacho, a cactus with leaves. AB - The specialized physiology of leafless, stem-succulent cacti is relatively well understood. This is not true, however, for Pereskia (Cactaceae), the 17 species of leafy trees and shrubs that represent the earliest diverging lineages of the cacti. Here we report on the water relations and photosynthesis of Pereskia guamacho, a small tree of the semiarid scrubland of Venezuela's Caribbean coast. Sapwood-specific xylem conductivity (Ksp) is low when compared to other vessel bearing trees of tropical dry systems, but leaf-specific xylem conductivity is relatively high due to the high Huber value afforded by P. guamacho's short shoot architecture. P. guamacho xylem is not particularly vulnerable to drought-induced cavitation, especially considering the high leaf water potentials maintained year round. This is confirmed by the lack of significant variation exhibited in Ksp between wet and dry seasons. In the rainy season, P. guamacho exhibited C3-like patterns of stomatal conductance, but during a prolonged drought we documented nocturnal stomatal opening with a concomitant accumulation of titratable acid in leaves. This suggests that P. guamacho can perform drought-induced crassulacean acid metabolism (CAM photosynthesis), although delta 13C values imply that most carbon is assimilated via the C3 pathway. P. guamacho leaves display very low stomatal densities, and maximum stomatal conductance is low whether stomata open during the day or night. We conclude that leaf performance is not limited by stem hydraulic capacity in this species, and that water use is conservative and tightly regulated at the leaf level. PMID- 17080641 TI - Comparison of leaf structure and photosynthetic characteristics of C3 and C4 Alloteropsis semialata subspecies. AB - Alloteropsis semialata (R. Br.) Hitchcock includes both C3 and C4 subspecies: the C3 subspecies eckloniana and the C4 subspecies semialata. We examined the leaf structural and photosynthetic characteristics of these plants. A. semialata ssp. semialata showed high activities of photosynthetic enzymes involved in phosphoenolpyruvate carboxykinase-type C4 photosynthesis and an anomalous Kranz anatomy. Phosphoenolpyruvate carboxylase; pyruvate, Pi dikinase and glycine decarboxylase (GDC) were compartmentalized between the mesophyll (M) and inner bundle sheath cells, whereas ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) occurred in both cells. A. semialata ssp. eckloniana also showed an anomalous non-Kranz anatomy, in which the mestome sheath cells included abundant chloroplasts and mitochondria. Rubisco and GDC accumulated densely in the M and mestome sheath cells, whereas the levels of C4 enzymes were low. The activity levels of photo-respiratory enzymes in both subspecies were intermediate between those in typical C3 and C4 plants. The values of CO2 compensation points in A. semialata ssp. semialata were within the C4 range, whereas those in A. semialata ssp. eckloniana were somewhat lower than the C3 range. These data suggest that the plants are C3-like and C4-like but not typical C3 and C4, and when integrated with previous findings, point to important variability in the expression of C4 physiology in this species complex. A. semialata is therefore an intriguing grass species with which to study the evolutionary linkage between C3 and C4 plants. PMID- 17080642 TI - A mutation affecting ASCORBATE PEROXIDASE 2 gene expression reveals a link between responses to high light and drought tolerance. AB - Molecular analyses of plants have revealed a number of genes whose expression changes in response to high light (HL), including the H2O2 scavenger, ASCORBATE PEROXIDASE 2 (APX2). We carried out a screen in Arabidopsis thaliana for lesions that alter HL-induced expression of APX2 to identify components in abiotic stress signalling pathways. High light was used as it can be instantaneously applied or removed and accurately measured. We identified a number of alx mutations causing altered APX2 expression. Here we describe the gain-of-function mutant, alx8, which has constitutively higher APX2 expression and higher levels of foliar abscisic acid (ABA) than wild type. In fact, exogenous ABA increased APX2 expression and the APX2 promoter contains ABA response elements. Furthermore, we have shown that HL stress increases ABA in wild-type plants, implicating ABA in the regulation of HL-inducible genes. The alx8 mutant is drought tolerant, exhibits improved water-use efficiency and a number of drought-tolerance genes are upregulated. Additionally, alx8 demonstrates the complexity of ABA-dependent and ABA-independent transcriptional networks as some components in both pathways are upregulated in alx8. This study provides evidence for common steps in drought and HL stress response pathways. PMID- 17080643 TI - The stimulating effects of ethylene and auxin on petiole elongation and on hyponastic curvature are independent processes in submerged Rumex palustris. AB - The flooding-tolerant plant species Rumex palustris (Sm.) responds to complete submergence with stimulation of petiole elongation mediated by the gaseous hormone ethylene. We examined the involvement of auxin in petiole elongation. The manipulation of petiolar auxin levels by removing the leaf blade, or by addition of synthetic auxins or auxin transport inhibitors, led to the finding that auxin plays an important role in submergence-induced petiole elongation in R. palustris. A detailed kinetic analysis revealed a transient effect of removing the auxin source (leaf blade), explaining why earlier studies in which less frequent measurements were taken failed to identify any role for auxin in petiole elongation. We previously showed that the onset of stimulated petiole elongation depends on a more upright petiole angle being reached by means of hyponastic (upward) curvature, a differential growth process that is also regulated by ethylene and auxin. This raised the possibility that both ethylene and auxin stimulate elongation only indirectly by influencing hyponastic growth. We show here that the action of ethylene and auxin in promoting petiole elongation in submerged R. palustris is independent of the promoting effect that these hormones also exert on the hyponastic curvature of the same petiole. PMID- 17080644 TI - Sanguinarine reductase, a key enzyme of benzophenanthridine detoxification. AB - Cultured cells of Eschscholzia californica respond to a yeast glycoprotein elicitor by producing benzophenanthridine alkaloids, which are excreted into the cell wall and the outer medium. These compounds, preferentially sanguinarine, are efficient phytoalexins because of their ability to intercalate double-stranded DNA (dsDNA), penetrate membranes and inhibit various enzymes containing SH groups. Externally added sanguinarine is rapidly taken up by intact cells and converted to dihydrosanguinarine, which is substituted intracellularly according to the biosynthetic route. A 29.5 kDa soluble enzyme that catalyses the reduction of sanguinarine and chelerythrine by either NADPH or NADH has been isolated and purified to homogeneity. Benzophenanthridines that accumulate in the outer medium, mainly 10-OH-chelerythrine, chelirubine and macarpine, are converted by the isolated enzyme and by intact cells at much slower rates than sanguinarine. The cellular capacity of uptake and conversion of sanguinarine largely surpasses the rate of alkaloid production. We conclude that the sanguinarine produced by intact cells, after excretion and binding to cell wall elements, is rapidly reabsorbed and reduced to the less toxic dihydrosanguinarine, which then undergoes further biosynthetic reactions. This recycling process would allow the presence of the toxic phytoalexin at the cellular surface without taking the risk of injuring the producing cell. PMID- 17080645 TI - Differential synthesis of phosphate-starvation inducible purple acid phosphatase isozymes in tomato (Lycopersicon esculentum) suspension cells and seedlings. AB - This study compares the influence of phosphate (Pi) deprivation on the coordinate synthesis of the principle Pi-starvation inducible (PSI) acid phosphatase (AP) isozymes in a suspension cell culture with the homologous in planta system. Tomato suspension cells express three PSI purple AP isozymes: a heterodimeric intracellular AP (IAP) composed of 63 and 57 kDa subunits, and two monomeric secreted APs (SAPs) (84 kDa SAP1 and 57 kDa SAP2) localized in the culture media. Immunoblots probed with rabbit antibodies raised against purified SAP1 or IAP indicated the immunological distinctiveness of SAP1 relative to IAP and SAP2. Time-course studies of cells and seedlings undergoing a transition from Pi sufficiency to Pi deficiency revealed a close relationship between total IAP or SAP activity and relative amounts of antigenic IAP or SAP polypeptides. Upregulation of the pre-existing IAP in 6-day-old Pi-deficient (-Pi) suspension cells coincided with a 20-fold reduction in intracellular free Pi levels, which occurred 2 d prior to initial accumulation of SAP1 and SAP2 in the culture media. Similarly, root-specific SAP synthesis in -Pi seedlings occurred at least 7 d following IAP induction in roots or shoots. Preferential sequestration of limiting Pi to the leaves of the -Pi seedlings was suggested by the delayed induction of leaf versus root IAP. Our results confirm recent transcript profiling studies suggesting that PSI proteins are subject to both temporal and tissue-specific syntheses in- Pi plants. PMID- 17080646 TI - Blood loss management in high-risk patients undergoing total knee arthroplasty: a comparison of two techniques. AB - Although it is well known that patients with preoperative hemoglobin levels <13.0 g/dL are at a higher risk for requiring postoperative transfusions, the ideal blood management strategy for this group of patients remains unclear. This study compared preoperative autologous donation with preoperative administration of epoetin alfa as a method to maximize perioperative hemoglobin levels and minimize blood transfusions in these high-risk patients undergoing total knee arthroplasty (TKA). Results show that both preoperative autologous donation and epoetin alfa were successful in decreasing the need for allogeneic blood transfusions following TKA in high-risk patients. Epoetin alfa was more effective in maximizing perioperative hemoglobin levels. PMID- 17080647 TI - Extramedullary femoral alignment instrumentation reduces blood loss after uncemented total knee arthroplasty. AB - The influence of different femoral alignment systems on blood loss and the need for blood transfusion after total knee arthroplasty (TKA) was studied. The blood loss was retrospectively recorded in two similar groups of consecutive patients. The first group consisted of 46 patients in whom TKA was performed using the intramedullary femoral alignment system and the second group consisted of 45 patients in whom the procedure was performed with the extramedullary system. In the intramedullary group, the mean volume of drained blood was 758 mL, whereas in the extramedullary group it was 613 mL (P < .05). More patients in the intramedullary. group required blood transfusions, but no significant difference was noted in the number of blood units transfused per patient. Extramedullary femoral alignment instrumentation reduces blood loss after cementless TKA. PMID- 17080648 TI - Use of multi-detector computed tomography for the detection of periprosthetic osteolysis in total knee arthroplasty. AB - This study determined the accuracy of plain radiography in detecting osteolytic lesions around total knee prostheses compared to multi-detector computed tomography (CT). Thirty-one patients diagnosed with periprosthetic osteolysis by multi-detector CT after total knee arthroplasty (TKA) were studied. Computed tomography for each patient was retrospectively reviewed in a blinded fashion. The plain radiographs for each patient that had been obtained prior to CT were reviewed in the same manner. The results of the CT were compared with the results of the radiographs. The number, size, and location of the lesions were compared. The multi-detector CT detected 48 lesions in 31 knees: 40 tibial lesions, 4 femoral lesions, and 4 patellar lesions. Radiographic diagnosis was made in 6 of the 40 tibial lesions, 2 of the 4 femoral lesions, and 0 of the 4 patellar lesions. Plain radiographs are inadequate for evaluating periprosthetic osteolysis in TKA with only 8 (17%) of 48 lesions detected by multi-detector CT visible on the standard radiographs. Multi-detector CT provides the surgeon with a diagnostic and preoperative planning tool when osteolysis is suspected. PMID- 17080649 TI - Prolonged storage of osteochondral allografts: does the addition of fetal bovine serum improve chondrocyte viability? AB - Osteochondral plugs were harvested from eight fresh human femoral condyles within 96 hours of donor death. The plugs were either stored in a serum-free media containing glucose, salts, and amino acids or 10% fetal bovine serum at 4 degrees C. After 28 days of storage, the osteochondral plugs were analyzed for chondrocyte viability and viable cell density using confocal microscopy, proteoglycan synthesis by (35)SO4 incorporation, and glycosaminoglycan content. Chondrocyte viability and cell density were significantly lower in grafts stored in serum-free media compared to fetal bovine serum, 27% versus 68% (P < .001) and 3250 cells/mm3 versus 8960 cells/mm3, respectively (P < .001). The metabolic activity determined by proteoglycan synthesis was significantly better in the specimens stored in fetal bovine serum (P < .01). No significant difference was detected between the glycosaminoglycan content in any of the specimens. These data suggest that the quality of osteochondral allografts as measured by chondrocyte viability, viable cell density, and proteoglycan synthesis is superior after storage in fetal bovine serum versus serum-free media. These results must be taken cautiously, however, as the clinical ramifications of storage in fetal bovine serum, including potential infectious disease transmission risks and immunogenic factors, have yet to be studied. PMID- 17080650 TI - Ehrlichiosis as a near-fatal cause of postoperative fever after bilateral total knee arthroplasty. PMID- 17080651 TI - Lateral retinacular release for anterior knee pain: a systematic review of the literature. PMID- 17080652 TI - Treatment of chondral defects in the patellofemoral joint. AB - Patellofemoral disease is one of the most controversial management issues in orthopedic surgery. Nonoperative management as a prerequisite first line treatment is successful in the majority of cases. However, a small subset of patients with persistent pain after adequate rehabilitation will be potential candidates for surgical intervention. Careful assessment of the underlying pathomechanics is critical for a successful outcome; these include malalignment of the extensor mechanism, trochlear dysplasia, soft-tissue imbalance, and chondral damage. As the pathology is multifactorial, the planning and treatment must be multifaceted. With careful patient selection, the options of titrated limited lateral release, restoration of MPFL function, tibial tubercle osteotomy, cartilage repair, and patellofemoral resurfacing provide improved functionality and pain relief for the young patient suffering from patellofemoral pain. PMID- 17080653 TI - Surgical options for treating injuries to the medial patellofemoral ligament. PMID- 17080654 TI - Reconstruction of the medial patellofemoral ligament for recurrent patellar instability. PMID- 17080655 TI - Patellofemoral forces after medial patellofemoral ligament reconstruction: a biomechanical analysis. PMID- 17080656 TI - [Expressions of Bcl-2 and Caspase-3 in the internal organs of rats when tetramine was administered]. AB - OBJECTIVE: To find whether Bcl-2 and Caspase-3 take part in the pathophysiological mechanism of tetramine toxification. METHODS: Sixty Sprague Dawley rats were divided into normal control group, the sham poisoned group, high dose poisoned group, low dose poisoned group. High dose poisoned group were administered 1.0 mg/kg weight body tetramine by mouth, however low dose poisoned group was administered tetramine 0.1 mg/kg weight body by mouth. The rats of the sham poisoned group were administered water, and rats of normal control group were given nothing. Bcl-2 and Caspase-3 were detected by immunohistochemistry staining and the results were assessed by image analysis system. RESULTS: The expressions of Bcl-2 and Caspase-3 in all organs were similar, ie, Bcl-2 and Caspase-3 expressed obviously in all organs of high dose poisoned group; in all organs of low dose poisoned group, they were hardly detected at 30 min after administration, however, at 3 h after administration, they could be detected obviously; Bcl-2 got to peak at 6 h-3 d after administration and Caspase-3 got to peak at 24 h-3 d after administration. CONCLUSION: Bcl-2 and Caspase-3 take part in the pathophysiological procedure of tetramine poisoned rats. PMID- 17080657 TI - [Changes of cyclin D1 expression following human brain contusion]. AB - OBJECTIVE: To study the relationships of Cyclin D1 expression with the posttraumatic intervals (PTI) following the cerebra, brainstem or cerebella contusion in human. METHODS: 88 cases of brain contusions of the closed head injury were investigated with pathological and Cyclin D1 immunohistochemistry methods. The results were analyzed by image analysis technique (IAT). RESULTS: The immunoreactivity of Cyclin D1 was almost disappeared in the core cells of the brain contusion. Cyclin D1-positive cells started to increase in the boundary of the brain contusion in the 1h group. Cyclin D1-positive cells were increased significantly in the 3 h-30 d groups and maintained at a high level in the boundary of the brain contusion of those groups. It is suggested that the Cyclin D1-positive cells were primarily origin from microglia and other glia. A few neurons expressed Cyclin D1. CONCLUSION: Cyclin D1 can express in several kinds of brain cells following the contusion, especially in the glia cells. Cyclin D1 positive cells were increased obviously and rapidly after injury, so it could be used as a reference marker for early stage brain injury. PMID- 17080658 TI - [Mechanism of acute liver injury induced by crushing hindlimbs in rabbits]. AB - OBJECTIVE: To investigate the role of oxidative stress in acute liver injury during crushing hindlimbs in rabbit. METHODS: The crushing injury model in rabbit was established by intermittent crushing the hind limbs of rabbit with standard weight. The ALT and AST activities were spectrophotometrically measured. The weight ratio (wet/dry,W/D) of livers was measured with scale, and the pathologic changes were observed. Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and total anti-oxidant capacity (T-AOC) as well as malondialdehyde (MDA) level were spectrophotometrically measured. RESULTS: As compared with control rabbits, crushing hindlimbs of rabbits induced acute liver injury with the increase in ALT and AST activities in serum,which were 4.31 (P < 0.01) and 10.54 times (P < 0.01) of control group respectively, there were cellular swellings and slight congestion of hepatic sinuses. In addition,crushing hind-limbs elicited significant decrease in SOD,CAT,GSH-Px activity and T-AOC to 17%, 29%, 24% and 21% (P < 0.01) compared with control group respectively, whereas MDA level markedly enhanced. CONCLUSION: Crushing hindlimbs of rabbits induced acute liver injury and significant decrease in anti-oxidant capacity, the latter maybe play an important role in crushing hind-limbs of rabbits-elicited the acute liver injury. PMID- 17080659 TI - [Measuring volume and density of human organs and its significance in medicine and forensic medicine: lung]. AB - OBJECT: To obtain values of normal human lung and diffuse pathological changes, in order to provide a simple and convenient diagnostic method for measuring human visceral organs in the autopsy of pathology and forensic pathology. METHODS: The exact mass, volume and density of normal and pathological lung were synchronously measured with the intelligentized volume-densimeter made by authors. RESULTS: The date-base about the volume and density of human lung with different races were established for anatomy, anthropology and biodynamics. In two cases of over-load in circulation and water toxicosis, the severe lung edema was proved with the scale of lung density, which was atypical edema in the lung tissue. CONCLUSION: Measurement of visceral organs in the autopsy can assist to an integrative pathological diagnosis. PMID- 17080660 TI - [Changes of adenylate cyclase on cerebral regions related to mophine dependence in rats]. AB - OBJECTIVE: To observe the changes of adenylate cyclase(AC) on cerebral regions related to morphine dependence in rats and investigate the relationship between the enzymological changes and the mechanism of morphine dependence. METHODS: The technique of enzyme-histochemistry was used to detect the variations of AC of special seven cerebral regions including frontalis cortex, lenticula, corpus amygdaloideun, substantia nigra, hippocampus, periaqueductal gray and locus coerleus in morphine dependent rats. The enzymological changes were observed by optical microscope. Changes of gray degree of these cerebral regions were also observed by using the image analysis system. RESULTS: Compared with those in control group, the contents of AC in morphine dependent groups were increased. CONCLUSION: The contents of AC are increase in those regions. The mechanism of morphine dependence close related to the increasing of AC. The correlation of the mechanism of morphine dependence and up-regulation of AC/cAMP-PKA system is discussed. PMID- 17080661 TI - [Changes of retinal ganglion cells and expression of Bad after optic nerve crush in rats]. AB - OBJECTIVE: To observe the change of retinal ganglion cells (RGCs)and the expression of Bad after optic nerve injury, so as to study the changes of optic function level on morphology and molecular. METHODS: The experimental models of optic nerve crush were established in fifty Wistar rats. At the different time after injuries (from one to twenty-eight day), the changes of RGCs were observed under microscope. Immunohistochemiscal technique and computer image analysis methods were performed to observe the changes of Bad in RGCs in rats. RESULTS: The number of RGCs was reduced significantly according to partial lesion of optic nerve crush. An initial loss of RGCs densities was accelerated in one week after nerve crush, two weeks later the trend mitigated. After four weeks, no obvious change were observed. The expression of Bad increased in 3 days, reached peak in 5 days, and declined one week later. No obvious changes were observed after two weeks. CONCLUSION: The expression of Bad lead to the loss of RGCs following optic nerve crush. This is the important reason of loss optic function. The identification on optic nerve injuries should be done at least four weeks later. PMID- 17080662 TI - [Computer simulation by passenger wound analysis of vehicle collision]. AB - OBJECTIVE: To reconstruct the course of vehicle collision, so that to provide the reference for forensic identification and disposal of traffic accidents. METHODS: Through analyzing evidences left both on passengers and vehicles, technique of momentum impulse combined with multi-dynamics was applied to simulate the motion and injury of passengers as well as the track of vehicles. RESULTS: Model of computer stimulation perfectly reconstructed phases of the traffic collision, which coincide with details found by forensic investigation. CONCLUSION: Computer stimulation is helpful and feasible for forensic identification in traffic accidents. PMID- 17080663 TI - [Change of nitric oxide in local muscle of crush injury hind-limbs in rats]. AB - OBJECTIVE: To investigate the change of nitric oxide (NO) level in local muscles induced by crushing hind-limbs in rats. METHODS: The rat experimental model of hind-limb crushing injury was established by crushing the hind limbs of rats with standard weight for 5 hours, thereafter releving the standard weight for another 5 hours. The rats were randomly divided into sham group, crushing group, crushing and injecting aminoguanidine (AG) group, crushing and injecting L-arginine (L Arg) group. The NOS activity and NO level in local muscles and serum were spectrophotometrically measured, and iNOS and eNOS protein expressions in local muscles were examined by immunohistochemistry. The weight ratio of wet to dry (W/D) of local muscles was measured and the pathologic changes were observed. RESULTS: The crushing hind-limbs induced serious primary and secondary injuries of local muscles such as rupture and rhadomyolysis of skeletal muscular fibers, interstitial vascular congestion and edema, and marked increase in W/D. The expressions of eNOS and iNOS were upregulated in local muscle in crush group compared with sham group. The NOS activity and NO level in local muscles and serum significantly increased. There was positive relationship between NO level and W/D in local muscles. With the usage of AG and L-arg, the hind-limb injuries seemed alleviated and aggravated, respectively. CONCLUSION: The crushing hind limbs of rats elicited the upregulation of eNOS and iNOS protein expression, the enhancement of NOS activity and the excess production of NO, the latter of which was involved in the mediation of secondary pathological changes in local muscles. PMID- 17080664 TI - [The study on relativity of the time of traumatic diskal hernia and the change of CT rate]. AB - OBJECTIVE: To find out the relativity between the time when the traumatic prolapse of intervertebral disc occurred and the change of the CT numerical value. METHODS: The CT numerical value of the vertebral pulp of herniation (denoted as A) and the CT numerical value of the vertebral pulp without herniation (denoted as B) were measured in for 198 patients and analyzed the relation between this parameter and the time when the prolapse of intervertebral disc occurred. RESULTS: It's found find out that there is relativity between the parameter and the time when the prolapse of intervertebral disc occurred, and this difference has the value for the statistics, when the parameter is at about 12 to -40 HU, the average value is -25.8 HU (A < B), it can be determined the prolapse of intervertebral disc occurred within 3 months ago. When the parameter is at about -13 to 0 HU, the average value is -6.6HU (A is similar with B), it can be determined the prolapse of intervertebral disc occurred at about 4-6 months ago. When the parameter is at about 37 to 289HU, the average value is 96.1 HU (A > B). It can be determined the prolapse of intervertebral disc occurred more than 12 months ago. CONCLUSION: The approximate time can be determined when the prolapse of interverterbral disc occurred by the paremeter. PMID- 17080665 TI - [RHCE genotyping in Chinese Han from north and Li from south]. AB - OBJECTIVE: To establish a method which processes RHCE genotyping with PCR. METHODS: Using PCR-SSP to detect RHCE genotype in 200 cases of Han population in north of China and Li population in south of China and detecting 5 samples of parentage testing at the same time. RESULTS: The results of RHCE genotyping in individuals of two populations are completely accorded with the results of serology typing. And the distribution of RH genotypic frequency in Han population in north China is: RHCCEE 1, RHCCEe 3, RHCCee 88, RHCcEE 4, RHCcEe 20, RHCcee 54, RHccEE 1, RHccEe 22, RHccee; The distribution of RH genotypic frequency in Li populatin in south China is RHCCEE 2, RHCCEe 2, RHCCee 106, RHCcEE 7, RHCcEe 62, RHCcee 10, RHccEE 3, RHccEe 8. The results of RHCE genotype detecting of parentage testing samples are accorded with the results of associated identification of 13 STR loci. CONCLUSION: PCR-SSP technology can exactly detect RHCE genotype in individuals of Han population in north China and Li population in south China. PMID- 17080666 TI - [Influence of meliorated AS-PCR primer for ABO genotyping]. AB - OBJECTIVE: To improve the correct rate of ABO genotyping by meliorating AS-PCR primer. METHODS: The primer P1 was changed into primer P1' by substituting the fifth base G for C of 3' end and the ABO genotyping results of primer P1 and P1' was compared and analysed. RESULTS: The non-specific product of OO typing is reducing and the wrong genotyping of OO and AO was avoided by meliorating AS-PCR primer. CONCLUSION: The rates of wrong ABO genotyping results could be effectively reduced by using altering primer P1'. PMID- 17080667 TI - [Genetic polymorphisms of 5 X-STR loci in Yunnan Nu population]. AB - OBJECTIVE: To investigate the allele and genotype frequencies of DXS6804, DXS6799, DXS8378, DXS7130 and DXS7132 in unrelated individuals of Nu population and establish the related genetic database. METHODS: Five X-STR loci were analyzed by PCR followed PAGE and silver staining. RESULTS: The allele frequencies of the five X-STRs in Yunnan Nu population are in accordance with Hardy-Weinberg equilibrium. CONCLUSION: Five X-STRs loci of Nu population could be used in forensic identification. PMID- 17080668 TI - [How to draw a conclusion in motherless parentage testing using short tandem repeats as genetic makers]. AB - OBJECTIVE: To calculate the exclusion power of STR loci in motherless parentage testing and to discuss how to draw a conclusion if there are inconsistent loci. METHODS: Based on the law of inheritance and allele frequency, the powers of exclusion of STR loci in motherless parentage testing (PE(M)) were calculated. Based on the mean PE(M) and mutation rate of 13 CODIS loci. The probabilities of inconsistence under paternity and non-paternity were calculated respectively according to binomial theorem. RESULTS: The PE(M) of locus having co-dominate alleles could be calculated as: PE(M) = (i = 1)sigma (n) p i 2(1-p (i))2+ (i < j)sigma (n) 2p (i)p (j)(1-p (i)-p (j))2. According to the formula, the average PE(M) of 13 CODIS was 0.411. Based on the mean PE(M) and mutation rate, the likelihood ratio of true father to random man (paternity index) was got using binomial theorem. CONCLUSION: The conclusion in motherless parentage testing could be drawn based on the likelihood ratio (paternity index) derived from mean PE(M) and mutation ratio. PMID- 17080669 TI - [Determination of scopolamine in the poisoning case by GC/MS]. AB - OBJECTIVE: To separate and determine scopolamine from the food in a poisoning case by GC/MS. METHODS: The scopolamine was determined by GC/MS/El used CP5860(CP sil8CB) column (30 mx 0.25 mmx 0.33 microm) with liquid- liquid extraction. RESULTS: The deny scopolamine was found in the case sample, and the chromatographic separation of the peaks is fine. CONCLUSION: The method is accurate and reliable. PMID- 17080671 TI - [Analysis on 55 cases of traumatic cerebral infarction in forensic identification]. AB - 55 cases of traumatic cerebral infarction collected from these 5 years were respectively studied to analyze its clinical manifestation, the characteristic of CT and MRI scan, the mechanism of illness, the degrees of injury, and to explore the relation of craniocerebral trauma and cerebral infarction. PMID- 17080670 TI - [The application of diminished criminal responsibility rating scale to schizophrenia offenders]. AB - OBJECTIVE: To explore the feasibility of Diminished Criminal Responsibility Rating Scale(DCRRS) to schizophrenia offenders. METHODS: The DCRRS were used respectively to 325 cases of schizophrenia offender which were divided into three groups according to the degree of criminal responsibility. RESULTS: There were significant differences in rating score among three groups (mild group 21.16 +/- 4.17, moderate group 25.87 +/- 5.43, major group 29.35 +/- 4.60), and all items have good correlation with the total score of the scale. 6 factors were extracted by the factor analysis, and their cumulative squared loadings is 68.485%. CONCLUSION: The diminished criminal responsibility in schizophrenia offenders could be divided into three grades. PMID- 17080672 TI - [Analysis on the injury by lock-against-theft]. AB - Nowdays, it is common seen that criminals tend to use the lock-against-theft for motorcycle to harm others as a tool. In order to find out the chief characters of this kind of injury, 28 cases in which victims were hurt by lock-against-theft for motorcycle were analyzed. In each case, the victims carefully questioned to confirm the damage procedure. Furthermore, the wound configuration was observed and the forming mechanism was deduced to provide morphological evidences for the judgement in this kind of case. PMID- 17080673 TI - [The clinical forensic medicine identification on pseudoseizures after head trauma]. AB - Two pseudoseizures after head trauma are reported. We summarize the clinical manifestation of pseudoseizures, and identify difference of pseudoseizures and Seizures after head trauma. The forensic identification of pseudoseizures after head trauma should base on the extend and position of cerebral trauma, sequelae focus in cerebral, and the clinical manifestation. PMID- 17080674 TI - [Delayed gastric emptying in 5 case]. AB - It is a common way for estimating of time of death in forensic science by the extent of gastric emptying, but this method may be less accurate in sometimes. Here, we report 5 cases with significant delayed gastric emptying and made discussion about it. PMID- 17080675 TI - [Condition and mechanism of formation of mummy in Shanghai region]. AB - To investigates the condition and the mechanism of the formation of mummy in Shanghai region. Three cases of mummy were normaly examinated. It showed that the formation condition of mummy were influenced by season, scene, and the corpse keeping methods etc. In warm and damp southern region in Shanghai, the mummy formation depended on lower temperature, dry environment season, dehydrated and hunger before death, and interfered factors like ventilating and turning over corpse frequently. PMID- 17080677 TI - [Anthropology play a key role in simulated portrait]. AB - The technology of simulated portrait played a key role in criminal cases' deterction recently, while anthropology is attached to it tightly. This paper analyzed and discussed the impact of anthropology in simulated portrait by means of studying the relationship between races, regions figures skeletons and physiognomy. PMID- 17080676 TI - [Analysis on 27 autopsy cases died of anaphylactic shock induced by mainline]. AB - 27 autopsy cases died of anaphylactic shock induced by mainline during 1996-2005 were selected and analyzed. The results showed that most anaphylactic shock induced by mainline were rapid type and more easily happened in individual clinique and lawless clinique, and it could be arisen by either antibiotics or non-antibiotics. In some cases, there may be no pathologic findings by autopsy. So, all the materials must be taken into account for determination the anaphylactic shock, such as drugs, clinic symptom, autopsy findings, and so on. PMID- 17080678 TI - [Method selection from medical imaging examinations in evaluation of pelvic injury in forensic medicine]. AB - OBJECTIVE: To table a proposal for select using the medical imaging methods to evaluate pelvic injury in forensic field, by studying the structure of pelvis and radiological methods in common use. METHODS: A study of several cases of pelvic injury was done, in which biomechanics and classification of pelvic injury were analyzed, moreover imaging methods were compared with each other, such as fluoroscopy, X-ray photography, computerized tomography (CT) and so on. RESULTS: Four cases of pelvic injury are all multiple injuries, confirmed by X-ray photography and CT examination approvingly. CONCLUSIONS: Authors advocate that pelvic injury mechanism and biomechanics should be considered accordingly, multiple injuries should be attracted notice, so as to select suitable imaging methods to evaluate pelvic injury exactly. PMID- 17080679 TI - [Development of a gas chromatography-mass spectrometry method for the quantification of propofol in blood]. AB - OBJECTIVE: A rapid and selective method has been developed for the determination of propofol in blood by gas chromatography-mass spectrometry (GC-MS). METHODS: The blood was extrasted with the solution of internal standard, the extracted residue was analyzed by gas chromatography-mass spectrometry using selected-ion monitoring mode. RESULTS: The linear range was 0.1-10 microg/mL. The coefficient(R2) was 0.993 1. The detection limit was 0.05 microg/mL. CONCLUSION: The method is simple and rapid, offering superior sensitivity and selectivity for propofol. The method can be used successfully during clinical and forensic toxicology. PMID- 17080680 TI - Four new macrocyclic trichothecenes from two strains of marine-derived fungi of the genus Myrothecium. AB - Three new macrocyclic trichothecenes, named 12'-hydroxyroridin E (1), roridin Q (2), and 2',3'deoxyroritoxin D (3), were isolated from the marine-derived fungus Myrothecium roridum TUF 98F42, and a new macrocyclic trichothecene, named roridin R (4), was isolated from Myrothecium sp. TUF 02F6 together with roridins A and H and isororidin E. The structures of new compounds were determined on the basis of their spectral data. Compound 2 possessed a unique ether moiety at the 13' position of 1. Compound 4 was a 2',3'-dihydro-2'-hydroxy derivative of roridin H. The IC50 values of compounds 1, 2, and 4 against the murine leukemia cell line L1210 were 0.19, 31.2, and 0.45 microM, respectively. Compound 3 showed antiyeast activity to Saccharomyces cerevisiae at 1 microg/disc (inhibition zone: 12.2 mm), which was about 10 time more active than roritoxin D (10.2 mm at 10 microg/disc). PMID- 17080681 TI - Total syntheses of (+)-1893B and its three diastereomers and evaluation of their biological activities. AB - The total syntheses of natural (+)-1893B (2) and three other diastereomers 14, 18, and 21 were accomplished. Starting from the sequential metathesis product 5 prepared in turn from a 7-oxanorbornene derivative (+)-4, 2 was synthesized by means of an epoxy-ring opening of 9a with trimethylsilylacetylide followed by Wacker-type oxidation of the resulting alkyne 10 for the construction of the gamma-lactone moiety. By applying the same synthetic sequence, three additional diastereomers of 2, 14, 18, and 21 were also synthesized. The biological activities of previously synthesized 1893A (1), 1893B (2), and the diastereomers of 1893B 14, 18, and 21 were investigated. PMID- 17080682 TI - Production of ansamycin polyketide precursors in Escherichia coli. AB - For the heterologous production of ansamycin polyketides such as rifamycin and geldanamycin in Escherichia coli, a number of unusual but important tools must be engineered into the bacterium. Here we demonstrate efficient production of the starter unit 3-amino-5-hydroxybenzoic acid (AHBA) and the methoxymalonyl extender unit in E. coli. Previous work has demonstrated the production of the ansamycin starter unit AHBA in E. coli in low yield. It was shown that the low yield is primarily due to acetylation of AHBA into N-acetyl-AHBA. Three methods for minimizing this side reaction were evaluated. First, a putative N-arylamine acetyltransferase (NAT) was deleted from the E. coli chromosome, although this did not alter N-acetyl-AHBA production. Next, E. coli grown in media devoid of glucose yielded a nearly equal mixture of AHBA and N-acetyl-AHBA. Lastly, the NAT inhibitor glycyrrhizic acid was shown to further inhibit the acetylation reaction. The entire set of genes for synthesizing the methoxymalonyl extender unit was transferred from the geldanamycin producer Streptomyces hygroscopicus into E. coli. The pathway specific ACP isolated from the resulting recombinant strain was found to predominantly occur as methyoxymalonyl-ACP. Together, these findings set the stage for engineered biosynthesis of ansamycin polyketides in E. coli. PMID- 17080683 TI - Geosmin biosynthesis in Streptomyces avermitilis. Molecular cloning, expression, and mechanistic study of the germacradienol/geosmin synthase. AB - Geosmin (1) is responsible for the characteristic odor of moist soil. The Gram positive soil bacterium Streptomyces avermitilis produces geosmin (1) as well as its precursor germacradienol (3). The S. avermitilis gene SAV2163 (geoA) is extremely similar to the S. coelicolor A3(2) SCO6073 gene that encodes a germacradienol/geosmin synthase. S. avermitilis mutants with a deleted geoA were unable to produce either germacradienol (3) or geosmin (1). Biosynthesis of both compounds was restored by introducing an intact geoA gene into the mutants. Incubation of recombinant GeoA, encoded by the SAV2163 gene of S. avermitilis, with farnesyl diphosphate (2) in the presence of Mg2+ gave a mixture of (4S,7R) germacra-1(10)E,5E-diene-11-ol (3) (66%), (7S)-germacrene D (4) (24%), geosmin (1) (8%), and a hydrocarbon, tentatively assigned the structure of octalin 5 (2%). Incubation of this germacradienol/geosmin synthase with [1,1-(2)H2] FPP (2a) gave geosmin-d1 (1a), as predicted. When recombinant GeoA from either S. avermitilis or S. coelicolor A3(2) was incubated with nerolidyl diphosphate (8), only the acyclic elimination products beta3-farnesene (10), (Z)-alpha-farnesene (11), and (E)-alpha-farnesene (12) were formed, thereby ruling out nerolidyl diphosphate as an intermediate in the conversion of farnesyl diphosphate to geosmin, germacradienol, and germacrene D. PMID- 17080684 TI - Tensidols, new potentiators of antifungal miconazole activity, produced by Aspergillus niger FKI-2342. AB - Two new furopyrrols, designated tensidols A and B, were isolated from the culture broth of Aspergillus niger FKI-2342 by solvent extraction, silica gel column chromatography and HPLC. Their structures were elucidated and shown to have the common skeleton of 6-benzyl-6H-furo[2,3-b]pyrrole. Tensidols A and B potentiated miconazole activity against Candida albicans. Tensidols also showed moderate antimicrobial activity only against Pyricularia oryzae. PMID- 17080685 TI - Isolation and identification of the icosalides--cyclic peptolides with selective antibiotic and cytotoxic activities. AB - Three cyclic peptolides have been isolated from two different fungal species and their structures determined. Icosalides A1 (1a), A2 (1b), and B (1c) each contain two serine and two leucine amino acid residues and incorporate two fatty acid moieties as part of the central twenty-member ring. 1a contains L-serine and both D- and L-leucine residues, while 1b and 1c contain only L-amino acid residues. Icosalide A1 displays antimicrobial activity against Streptococcus pyogenes, S. pneumoniae (Felton), and Enterococcus faecalis. Icosalides A2 and B are cytotoxic to replicating MDCK cells. PMID- 17080686 TI - Cylindrocyclin A, a new cytotoxic cyclopeptide from Cylindrocarpon sp. AB - In the course of a screening of fungal extracts for new metabolites with cytotoxic activities cylindrocyclin A (1) was isolated. The producing strain was identified as Cylindrocarpon sp. by microscopy and ITS rDNA sequence analysis. 1 is a novel compound that exhibits cytotoxic acticity against six different cell lines with IC50 values ranging from 11 to 53 microM. 1 has no antibacterial or antifungal activity. The compound is a cyclic nonapeptide comprising three alanines, five leucines and one isoleucine. Four amino acids are N-methylated. Its structure was elucidated by spectroscopic methods. PMID- 17080687 TI - Sporminarins A and B: antifungal metabolites from a fungicolous isolate of Sporormiella minimoides. AB - Cultures of a fungicolous isolate of Sporormiella minimoides afforded two new polyketide metabolites which we have named sporminarins A (1) and B (2). The planar structures of 1 and 2 were elucidated by analysis of NMR and MS data, and by chemical methods. 1 exhibited significant antifungal activity against Aspergillus flavus. PMID- 17080688 TI - 1-Deoxynojirimycin derivatives from the marine sponge Lendenfeldia chondrodes. AB - Two 1-deoxynojirimycin derivatives, 1-deoxynojirimycin-6-phosphate (1) and N methyl-1-deoxynojirimycin-6-phosphate (2) were isolated from an aqueous extract of Micronesian marine sponge Lendenfeldia chondrodes for the first time as natural products. Structures of these compounds were assigned on the basis of their spectral data and chemical degradation. PMID- 17080689 TI - Amphidinin B, a new polyketide metabolite from marine dinoflagellate Amphidinium sp. AB - A new polyketide metabolite, amphidinin B (1), has been isolated from a marine dinoflagellate Amphidinium sp. and the structure and absolute stereochemistry were elucidated on the basis of spectroscopic data and chemical means. PMID- 17080690 TI - Fasting from midnight--the history behind the dogma. AB - The purpose of this chapter is to review historical fasting guidelines and how the dogma of fasting from midnight arose and came to be challenged by randomized clinical trials of preoperative clear liquids versus overnight fast. Medical and anaesthesia textbooks and journals from the 19th and 20th centuries were consulted, and the results of clinical trials and the reaction to them are reviewed. The dogma appeared to result from extrapolation of pulmonary aspiration risk in 'full-stomach' emergency cases to healthy elective cases. This was reinforced when 25 mL in the stomach, present in half of all healthy fasting patients, was used as a surrogate marker for high risk of aspiration. Subsequent large-scale studies showed the risk to be minimal. Meta-analysis of randomized clinical trials demonstrated the safety of clear oral liquids until 2 hr preoperatively in healthy patients undergoing elective surgery. Reaction was cautious but led to eventual acceptance of evidence-based fasting guidelines. PMID- 17080691 TI - The resistance to changing guidelines--what are the challenges and how to meet them. AB - Bridging the gap between scientific evidence and its practical application is of the utmost importance in improving the quality of care and increasing patient safety. Guidelines based on evidenced-based medicine (EBM) have led to improved performances and better outcomes. However, even though scientific data are available, resistance to adopting evidence-based guidelines is still enormous. Significant barriers hinder the introduction of best medical practice into the daily clinical routine. The barriers to implementing change are complex, multifunctional, and influenced on many levels by various interests both inside and outside the health-care system. Political, organizational, financial, cultural and scientific interests are regarded as being as important as the perception of patients and health workers. Strategies need to be planned which take account of the multidimensional character of quality of care and incorporate it at the various levels. The conclusion, therefore, is that we need to combine methods and tools to tailor our interventions to the patient's needs. PMID- 17080692 TI - The physiology of gastric emptying. AB - The gastric emptying rate is a carefully regulated process consisting of different mathematically defined phases. The gastric metabolic load, as well as neural regulatory mechanisms and hormonal influences, cooperate in order to achieve a well-balanced emptying of contents from the stomach into the duodenum for absorption in the small intestine. This finely tuned regulation is primarily regulated by the release of gastrointestinal peptide hormones which serve to counteract the emptying process in the fed state and to stimulate sweeping contractions in the fasted state, most likely in order to prepare the stomach for another meal. We have found that the two peptide hormones ghrelin and glucagon like peptide- I (GLP- I) have a great impact on the regulation of gastric emptying: ghrelin is a most potent stimulator of gastric contractions and emptying, and GLP- I profoundly inhibits this emptying process. These data suggest possibilities for governing the rate of gastric emptying as a natural step in achieving metabolic balance and control. PMID- 17080693 TI - Management of pulmonary aspiration. AB - Pulmonary aspiration of gastric contents in the perioperative phase is associated with increased postoperative morbidity and mortality. For the management of aspiration, differentiation between acid-associated aspiration pneumonitis and aspiration pneumonia as a consequence of a secondary bacterial contamination is of crucial importance. The incidence of aspiration in elective surgery is 1 per 2000-3000 anaesthesias in adults. In children, it is slightly more common with 1 per 1200-2600 anaesthesias. In the context of emergency anaesthesias the incidence of aspiration is three to four times higher. The risk particularly increases with recent ingestion of solid food or fluids, with older patients, with pregnant women, and with consciousness-reduced patients. Besides giving a review of the pathophysiology, incidence, and the risk factors of aspiration, this article places emphasis on the practical management of this anaesthesia associated complication. Cricoid pressure, as a non-evidence-based but clinically wide-spread method in the context of the prophylaxis of aspiration, is discussed critically. The main part deals with strategies to structure the management of aspiration by use of scientific concepts based on medical crisis management. For this, an algorithm based on current scientific investigations is presented. PMID- 17080694 TI - The metabolic effects of fasting and surgery. AB - This review discusses the metabolic changes that occur during fasting, especially in relation to surgical stress. Currently available evidence on the relevance of different aspects of perioperative nutritional support are presented. Attenuation of surgical stress and insulin treatment in insulin-resistant patients after surgery and trauma have been shown to markedly affect stress metabolism as well as patient morbidity and mortality. Avoiding preoperative fasting has been shown to be related to a substantial reduction in postoperative stress and insulin resistance. Implementation of a standardized programme with a multimodal approach using evidence-based perioperative routines has been shown to further reduce surgical stress and to markedly enhance functional recovery also after major surgical procedures. PMID- 17080695 TI - How perioperative fluid balance influences postoperative outcomes. AB - Fasting, anaesthesia and surgery affect the body's physiological capacity not only to control its external fluid and electrolyte balance but also the internal balance between the various body fluid compartments. Conversely, abnormalities of fluid and electrolyte balance may adversely affect organ function and surgical outcome. Perioperative fluid therapy has a direct bearing on outcome, and prescriptions should be tailored to the needs of the patient. The goal of fluid therapy in the elective setting is to maintain the effective circulatory volume while avoiding interstitial fluid overload whenever possible. Weight gain in elective surgical patients should be minimized in an attempt to achieve a 'zero fluid balance status'. On the other hand, these patients should arrive in the anaesthetic room in a state of normal fluid and electrolyte balance so as to avoid the need to resuscitate fluid-depleted patients in the anaesthetic room or after the induction of anaesthesia. Optimal fluid delivery should be part of an overall care package that involves minimization of the period of preoperative fasting, preoperative carbohydrate loading, thoracic epidural analgesia, avoidance of nasogastric tubes, early mobilization, and early return to oral feeding, as exemplified by the enhanced recovery after surgery programme. PMID- 17080696 TI - The evidence base behind modern fasting guidelines. AB - Fasting before general anaesthesia aims to reduce the volume and acidity of stomach contents, thus decreasing the risk of regurgitation/aspiration. The objectives of the Cochrane report which are summarised in this paper, were to systematically review the effect of different preoperative fasting regimes (duration, type and volume of intake) on perioperative complications and patient wellbeing. Few trials reported the incidence of aspiration/regurgitation or related morbidity but relied on indirect measures of patient safety ie. intraoperative gastric volume and pH. There was no evidence to suggest intake of fluids up to 2 hr preoperatively has an impact on patients gastric volumes or pH. Intake of fluids up to 90 min preoperatively had no impact on gastric contents but this was based on small numbers. In addition, permitting patients to drink water preoperatively resulted in significantly lower gastric volumes. Clinicians should evaluate this evidence for themselves and when necessary, adjust existing fasting policies. PMID- 17080697 TI - Modern fasting guidelines in children. AB - Pediatric fasting guidelines are intended to reduce the risk of pulmonary aspiration of gastric contents and facilitate the safe and efficient conduct of anesthesia. Recent changes in these guidelines, while assuring appropriate levels of patient safety, have been directed at improving the overall perioperative experience for infants, children, and their parents. Now after nearly 15 years of practice worldwide, the relative safety and benefits of allowing clear liquids up to 2 hr prior to anesthesia for otherwise healthy children are well established. Shortened fasting periods for breast milk (3 hr), formula (4 hr) and light meals (6 hr) are supported by accumulated experience and an evolving literature that includes evidence of minimal gastric fluid volumes (GFVs) at the time of surgery. Ideal fasting intervals for children with disorders that may affect gastrointestinal transit have yet to be determined. PMID- 17080698 TI - Modern preoperative fasting guidelines: a summary of the present recommendations and remaining questions. AB - This chapter is complementary to the others in this volume focusing on preoperative fasting routines. In it we discuss some of the issues in need of more research to define best practice. One of these is the role of fasting in emergency patients. Modern preoperative fasting recommendations almost exclusively deal with elective patients. In emergency patients preoperative fasting cannot secure gastric emptying to reduce the risk of pulmonary aspiration. Hence, surgery should be timed according to the urgency of the situation, and the patient should always be treated as if the stomach was full. More data are needed to better define what is going on in the gastrointestinal tract during the perioperative period in these patients. In certain patient groups--such as patients with diabetes, gastro-oesophageal reflux disease and/or obesity--the data are insufficient to give complete guidance to best practice. Preoperative fasting guidelines also affect fluid balance and perioperative fluid management, a topic of debate in recent years. In addition, carbohydrate-enriched fluids for oral use in the preoperative phase have been shown to have a positive effect on postoperative metabolism. Recent studies also suggest that the immune system would be less affected by surgery with such preparations. Last but not least, new scientific evidence alone is not enough to change daily practice. Active implementation of new evidence is also needed. To improve perioperative care, anaesthesiologists, surgeons and the nursing staff must work together. PMID- 17080699 TI - On the construction of clinical life table for studying the survivability of diagnosed AIDS cases. AB - In this paper, an attempt has been made to estimate the various survival functions of clinically diagnosed AIDS cases by the life table method. The actuarial life table method has already been applied to clinical data for many decades. To exhibit computations for the clinical life table, we have used the clinical data gathered on diagnosed 731 male and female AIDS patients recorded from Dec.1993 up to Dec.2002 in Delhi. The method is also used to study the nature of the survival functions for the AIDS patients with different causes of death. The comparisons among various survival functions are shown with the help of the graphs. PMID- 17080700 TI - Entomological indices of Aedes aegypti at some international airports and seaports of southern India--a report. AB - Entomological surveys were undertaken at some of the international airports/seaports (Bangalore, Calicut, Chennai, Cochin, Thiruvanathapuram and Vishakapatnam) to find out the breeding prevalence of dengue vector mosquito in diverse breeding containers from 1998 to 2004. Three vector indices (House index, Container index and Breateu index) were used to assess the breeding potential at each airport/seaport. International Health Regulations urged national governments to keep all the international airports/seaports and peripheral areas up to 400 meters free from Aedes aegypti mosquitoes. However, surveys revealed high vector indices at all the airports and seaports. Health authorities of airports/seaports need to take cognizance of these facts and develop action plan for appropriate control measures with emphasis on vector surveillance. PMID- 17080701 TI - Evaluation of malaria endemicity by peptide ELISA. AB - Interest in mapping the malaria transmission has motivated a need to develop a simple serological assay in a defined population. Evaluation of coded samples by peptide ELISA provided a framework to estimate the malarial impact. Comparison of field data and ELISA OD values in different malariogenic areas shows how the disease has been more correctly interpreted. Here we provide an empirical approach to estimate Annual Parasite Index (API) as well as Equivalent Transmission Index (ETI) using a combination of epidemiological, parasitological and immunological data. We estimate that there were 3 different malariogenic status like low, moderate and high transmission zones based on their' ELISA OD values. Our indigenous developed ETI estimates are 10 fold higher than API reported by the Primary Health Centre. Our record indicates that incidences of malaria will continue to be underestimated unless we adapt an alternative strategy. In order to verify the scope of malaria surveillance, coded samples were tested. Comparison of ELISA OD, API and ETI of the coded samples indicated Rourkela had high, Shahjahanpur had moderate and Bangalore had the lowest malaria transmission. For mass blood slide examination, microscopic method is a tedious process prone to human error while largely automated ELISA could reduce the scope for human error and could be a supplement for microscopic process. PMID- 17080702 TI - Preservation of Leishmania donovani promastigotes in blood agar slants. AB - Long-term cultivation of Leishmania promastigotes by weekly passage to fresh medium was reported to be disadvantageous because needs labor, risk of contamination, lowering in infectivity and virulence pattern. Cryopreservation and Lyophilization require expensive facilities which could be a burden and unaffordable to most laboratories of developing countries where the disease is endemic. These problems could be minimized by simple preservation of Leishmania donovani promastigotes in blood agar slants at 7-8 degrees C for 6-7 months. The preserved promastigotes were examined for viability up to one year at a regular interval of one month. Viable promastigotes were found and revived successfully from all the slants stored up to 7 months after that, the viability of promastigotes was found to be decreased in the slants of 8-9 month storage. No viable promastigotes were recovered from the slants stored up to 11-12 months. By this method, the promastigotes can easily be stored up to 7 months without loss of biological activity. The number of passage of promastigotes to fresh medium has been greatly reduced by this method from 30 times to 01 when compared with weekly passage in liquid medium. This simple and economical method can be recommended for short storage of Leishmania culture without loss of any activity. PMID- 17080703 TI - Operationalisation of surveillance of communicable diseases in Chandigarh. AB - Weekly reports of listed communicable diseases from various departments and centres of Government Medical College, Chandigarh, involved in clinical care and laboratory diagnosis, compiled and forwarded by Department of Community Medicine, sub-nodal centre under National Surveillance Programme for Communicable Diseases (NSPCD), to Anti- Malaria-cum-Nodal Officer, NSPCD were analysed for a period of one year Out of 14,082 cases of various communicable diseases 9166 (64.62%) were of Acute Respiratory Infections (ARI), 3586 (25.78%) of Acute Diarrhoeal Diseases (ADDs) and 576 (4.10%) of Pulmonary Tuberculosis. The proportion of ARI appeared higher among females while that of other diseases was higher among males. Most cases of ARI (76.5%) and Pneumonia (3.09%) reported in winter, ADDs (38.89%) and Pulmonary Tuberculosis (4.68%) in summer and Typhoid (1.57%) and Viral Hepatitis (1.23%) in monsoon season. No significant gender predilection was seen. Overall reporting of communicable diseases seen to be significantly more during winter and summer compared to monsoon season, with specific seasonal trends demonstrated by various morbidities. PMID- 17080704 TI - Acute lower respiratory tract infection among under- fives in urban eastern India -an appraisal of risk factors. AB - 200 under-fives children diagnosed as having ALRTI in pediatric OPD and indoor ward were sampled using standard case definition, and bodyweight was noted for each. All relevant factors of ALRTI were studied using pre-designed and pre tested proforma. 90 (45%) were underweight and 143 (71.5%) had >5 episodes of ALRTI. Socio-economic status was inversely related to the percentage having >5 ALRTI episodes. 93.0% of children whose parents were up to primary education level suffered from >5 episodes of ALRTI as compared to 55.3% whose parents had higher literacy levels (Z=7.0, P<0.05). Odds ratio for absence of exclusive breastfeeding (14.1), absence of measles vaccination (6.1), attached kitchen (6.9) were very high. PMID- 17080705 TI - Decline of brugian filariasis in Cherthala taluk, Alappuzha district, Kerala. AB - A total of 4492 persons from 5 panchayats and 1 town were investigated from the Brugia malayi most endemic taluk of Cherthala, Alappuzha district of Kerala state. The urban area in Cherthala taluk only revealed mf carriers; mf rate was 0.13%. Rural areas in Cherthala taluk were free from infection. Microfilaria rate had declined by 99.5% and disease rate by 90.7% in Cherthala compared to 1934 prevalence. Shedding of sheath by B. malayi microfilariae was recorded for the first time in India. The youngest person with microfilaria and disease manifestation was 4 1/2 and 9 years respectively. All the 3 major vectors, Mansonia annulifera, Ma.uniformis and Culex quinquefasciatus were prevalent throughout. Complete disappearance of brugian filariasis from this taluk is a distinct possibility. The reasons for the drastic decline are discussed. PMID- 17080706 TI - Hypogonadism in male Leprosy patients--a study from rural Uttar pradesh. AB - Hypogonadism in male patients with Leprosy is common and may identify patients with future risk for bone loss and osteoporosis. In the present study, we evaluated gonadal function in 71 male patients with Leprosy both clinically and by estimation of serum testosterone levels. The patients belonged to selected rural areas of Uttar pradesh, with majority aged less than 50 yrs (74.6%), Hindus (66.7%), illiterate (60.9%), and of low socioeconomic status (58% with per capita income < Rs.500 per month). Most patients had multibacillary Leprosy (83.1%), duration less than 2 years (75.4%) and had received antileprosy drugs for less than a year (95.6 %). Seven patients (9.9%) had clinical features of hypogonadism such as gynaecomastia, decreased sexual hair and infertility. Serum testosterone levels, estimated in 31 of the patients, revealed low values in 25.8% (8/31) patients (Mean 4.65+/-3.37 ng/ml). Age, duration of Leprosy and socioeconomic status but not type of Leprosy or treatment duration affected hypogonadism significantly. The results of the present study indicate a high frequency of hypogonadism among rural male Leprosy patients that warrants routine screening to identify patients at risk for osteoporosis and possible prevention with testosterone replacement therapy. PMID- 17080707 TI - Microbiological profile of neonatal septicemia in a pediatric care hospital in Delhi. AB - Septicemia continues to be an important cause of morbidity and mortality in the neonatal units and periodic review of cases to assess any changing trends in the infecting organisms and their antimicrobial susceptibility is important. Over a period of one year (July 2000 to June 2001), 632 samples of blood cultures were submitted to the bacteriology laboratory Microbiology, Lady Hardinge Medical College. These samples were investigated for microbial etiology and the isolates obtained were tested for their susceptibility to the commonly used antibiotics. Twenty per cent (125) cases were culture positive. Gram-negative bacteria were the predominant isolates (62%), commonest being Klebsiella pneumoniae (34%) followed by E. coli (17%), Acinetobacter spp. (9%) and Enterobacter aerogenes (2%). Gram-positive cocci were isolated in 20% cases, of which coagulase negative staphylococcus was the predominant isolate (11%) followed by Enterococcus spp. (5%) and S. aureus (4%). Candida spp. was isolated from 18% of cases. Resistance to commonly used antibiotics was seen in more than 35% of isolates. An alarming observation was the very high incidence of resistance to amoxycillin+clavulanic acid and ceftriaxone (>80%). All isolates showed highest susceptibility to ciprofloxacin. PMID- 17080708 TI - Seroprevalence of Chalmydia trachomatis infection amongst patients with pelvic inflammatory diseases and infertility. AB - Chlamydia trachomatis (C. trachomatis) is a common cause of pelvic inflammatory disease in female population. This infection can be diagnosed both by culture and serology. The present study evaluated the seroprevalence of chlamydial infection in patients with pelvic inflammatory disease (ND) and infertility and in control population of healthy normal females. The seroprevalence was found to be 82.7% in patients and 32% in controls by Enzyme Linked Immuno Sorbent Assay (ELISA) for any one or more class of antichlamydial antibodies (IgG/IgM/ IgA). The study demonstrates the importance of serology to monitor trends of chlamydial infections in women with PID and infertility. PMID- 17080709 TI - Comparative analysis of intestinal parasitic infections in slum, rural and urban populations in and around union Territory, Chandigarh. AB - A study was conducted in and around the union Territory, Chandigarh, India to estimate the prevalence of intestinal parasitic infections in different population groups. The stool samples from a total of 600 subjects selected by cluster sampling technique were examined by direct microscopy. The prevalence of intestinal parasitic infections was found to be 14.6% with highest prevalence of 19% from the slum area. Children were the most commonly affected group (18%) with those from slums showing the highest prevalence (24.6%). The most common parasite was Giardia (5.5%) followed by H. nana and Ascaris (2.8% and 2.7% respectively). PMID- 17080710 TI - Effectiveness of modular training regarding Revised National Tuberculosis Control Programme among paramedical workers in Delhi. PMID- 17080711 TI - Sexual risk behaviors in HIV positive individuals in north India. PMID- 17080712 TI - Age-wise seroprevalence of hepatitis B infection in clinical cases of jaundice attending a tertiary health care institute of Delhi. PMID- 17080713 TI - Chryseobacterium indologenes bacteremia: a case report. PMID- 17080714 TI - Chaperone and anti-chaperone: two-faced synuclein as stimulator of synaptic evolution. AB - Previous studies have shown that beta-synuclein (beta-syn), the homologue of alpha-syn, inhibited alpha-syn aggregation and stabilized Akt cell survival signaling molecule, suggesting that beta-syn was protective against alpha-syn related neurodegenerative disorders, such as Parkinson's disease and diffuse Lewy body disease. However, emerging evidence argues that the situation may be not so simple. Two missense mutations of beta-syn were identified in familial and sporadic diffuse Lewy body disease, and wild type beta-syn was induced to form fibril structures in vitro, while, alpha-syn was shown to be protective against neurodegeneration caused by deletion of cysteine-string protein-alpha, the presynaptic cochaperone to Hsc70 in mice. Collectively, alpha- and beta-syn are both, but in varying degrees, featured with two opposite properties, namely normal chaperone and anti-chaperone. By reviewing recent progress in syn biology with a particular focus on beta-syn, this manuscript refers to the intriguing possibility that the dual syn proteins might have acquired a driving force for synaptic evolution. Hypothetically, the anti-chaperone syn may provoke stress induced diverse responses, whereas, the chaperone syn may provide buffering for them, allowing accumulation of nonlethal phenotypic variations in synapses. Consequently, dual syn proteins may cope with forthcoming stresses in the brain by stimulating adaptive evolution. In this context, failure to regulate this process due to various causes, such as gene mutations and environmental risk factors, may result in imperfect adaptability against stresses, leading to neurodegenerative disorders. PMID- 17080715 TI - Quantitative mRNA expression analysis of neurotrophin-receptor TrkC and oncogene c-MYC from formalin-fixed, paraffin-embedded primitive neuroectodermal tumor samples. AB - Most recent studies analyzing candidate biological prognostic factors (including neurotrophin receptor TrkC and proto-oncogene c-MYC) in childhood primitive neuroectodermal brain tumors (PNET) are limited by small patient numbers due to dependence on fresh-frozen tumor material. In contrast, large archives of formalin-fixed, paraffin-embedded PNET samples exist from homogeneously treated patients. The ability of real-time RT-PCR to assay very small mRNA fragments makes this assay amenable to studies where the RNA is moderately or even highly degraded. We have optimized RNA isolation from archive PNET samples and found that TrkC and c-MYC mRNA measurements significantly correlated with those obtained from matching fresh-frozen tissues. Exploitation of already existing archives of formalin-fixed paraffin-embedded PNET samples may accelerate the building of better stratification systems for PNET patients. PMID- 17080716 TI - Olfactory neuroepithelioma: an immunohistochemical and ultrastructural study. AB - Three cases of olfactory neuroepithelioma are presented in this report. Histologically, these tumors were composed of small cells with round to oval, relatively hyperchromatic nuclei and scanty cytoplasm. The tumor cells were occasionally observed in tubular formations or rosette-like arrangements. Immunohistochemically, the tumor cells showed a positive reaction for cytokeratin AE1, cytokeratin CAM5.2, Ber-EP4, antisynaptophysin and anti-S100 protein in all cases. In two cases, LH-RH was detected in the tumor cells. Ultrastructurally, the tumor cells had the differentiation features of olfactory epithelium. Olfactory neuroepithelioma is a rare occurrence and it can be very difficult to distinguish olfactory neuroepithelioma from small cell carcinoma, neuroendocrine carcinoma and so-called "olfactory neuroblastoma" on the basis of hematoxylin and eosin stained sections alone. In controversial cases, a diagnosis of olfactory neuroepithelioma must be substantiated by ultrastructural and immunohistochemical findings, particularly regarding the detection of Ber-EP4 and LH-RH immunoreactivity. PMID- 17080717 TI - Oligonucleotide DNA chips are useful adjuncts in epigenetic studies of glioblastomas. AB - Several studies have suggested that hypermethylation and hypomethylation of CpG islands within the promoters and 5' exons of tumor-related genes are closely associated with carcinogenesis. However, large-scale analysis of candidate genes has been hampered by the lack of a high throughput approach for analyzing methylation patterns. Using methylation-specific oligonucleotide (MSO) chips, we evaluated the methylation patterns of eight samples of fresh frozen glioblastoma tissue. The MSO chip used contained DNA probes with the CpG sites of p16 (p16INK4A, CDKN2A), MGMT (O6-Methylguanine-DNA-methyltransferase), APC (adenomatous polyposis coil), RASSF1A (human RAS effect homolog), which are usually hypermethylated in cancer cells and MAGE (melanoma antigen), which is usually hypomethylated in cancer cells. We selected CpG sites for analysis; 28 CpG sites (263 bp) for p16, 26 CpG sites (249 bp) for MGMT, 16 CpG sites (195 bp) for APC, 22 CpG sites (262 bp) for RASSF1A and 18 CpG sites (235 bp) for MAGE. We then constructed primer sets not including CpG sites. Bisulfite modification of genomic DNA, methylation specific PCR, hybridization and image scan with data analysis and sequencing of the bisulfite modified DNA were carried out. Of the eight glioblastomas, hypermethylation of the 5'-CpG sites of the MGMT were found in two, RASSF1A were found in five, and p16 and APC genes were not found in any cases and hypomethylation of that of the MAGE was found in eight cases. These results obtained from the oligo DNA chip study were correlated well with the sequencing data of bisulfite modified genomic DNA except in regard to the RASSF1A and MAGE genes. The devised MSO DNA chip is a useful tool for studies on methylation. PMID- 17080718 TI - Heterogeneous inclusions in neurofilament inclusion disease. AB - Neurofilament inclusion disease (NFID) is a rare disease, whose pathogenesis remains to be elucidated. Immunoreactivity of ubiquitin-binding protein p62 has been reported in various neurodegenerative diseases, but it has not been studied in NFID. In this report we show p62 immunoreactivity in neuronal perikaryon of three cases of NFID. We found inclusions in NFID to be heterogenous based on immunoreactivity for alpha-internexin, phosphorylated neurofilament-H, p62 and ubiquitin. Moreover, we showed both p62- and alpha-internexin-immunoreactive inclusions within the perikarya of the same neuron. Electron microscopy findings support the notion that inclusions in NFID are heterogenous. The present study extends the list of proteins that have been identified as components of neuronal inclusions in NFID, and may help account for the pathogenesis of NFID. PMID- 17080719 TI - Expression of cyclo-oxygenase-2 in ependymal tumors. AB - Up-regulation of cyclo-oxygenase-2 (COX-2), a cytokine-induced enzyme that metabolizes arachidonic acid into prostaglandins, has been described in some brain tumors, including astrocytomas. Little is known about its expression in ependymal neoplasms. The objective of the present study was to assess COX-2 immunostaining of ependymal tumors. Retrospective COX-2 immunohistochemical analysis was conducted on 117 ependymal tumors. Statistical analysis was performed using Student t-test. The study group (56 men and 44 women, mean age, 30.8 years) was comprised of 48 low-grade ependymomas (WHO grade II), 12 anaplastic ependymomas (WHO grade III), 27 myxopapillary ependymomas (WHO grade I) and 13 subependymomas (WHO grade I). At last known follow-up (range, 12-226 months; mean, 74 months), 52 patients were alive with no evidence of tumor, 16 patients were alive with residual tumor, nine patients died with tumor, one patient died with no tumor and three died with tumor status unknown. Nineteen patients had less than 12 months of follow-up. Thirty-six (36%) patients had tumors, which demonstrated positive COX-2 staining, including 16/27 (59%) myxopapillary ependymomas, 3/13 (23%) subependymomas, 14/48 (29%) ependymomas and 3/12 (25%) anaplastic ependymomas. Statistically significant COX-2 positive immunostaining was observed in myxopapillary ependymomas versus WHO grade II (P = 0.03) and grade III (P = 0.02) tumors. Increased COX-2 expression in myxopapillary ependymoma as compared to the WHO grade II and II ependymoma was observed. The reason for this apparent increased immunoexpression in these low grade tumors is uncertain. COX-2 inhibitors may play a role in treatment of the subset of ependymal tumors that demonstrate increased expression. COX-2 staining did not reliably predict tumor behavior. PMID- 17080720 TI - Report on the first Chinese family with Gerstmann-Straussler-Scheinker disease manifesting the codon 102 mutation in the prion protein gene. AB - The authors found a female patient aged 33-years with dementia and cerebellar ataxia rapidly progressing for a year. EEG tracings were abnormal but without features of typical CJD. The patient died 13 months after the onset of illness. Biopsy of her cerebral cortex showed moderate spongiform changes, neuronal loss and gliosis. Numerous deposits of eosinophilic substance amorphous or in the shape of Kuru plaques were disclosed in the cerebral cortex. All deposits stained strongly with monoclonal 3F4 antibody to human prion protein. Genetic studies disclosed the Pro to Leu point mutation at codon 102 with a 102 Leu-129 Met in the PrP gene. Codon 129 was heterozygous for Met/Val, and codon 219 was homozygous for Glu/Glu. It was established; moreover, that the patient's grandfather had a similar disease and died at age 48 and the patient's brother died after a 10-year long neurological disease diagnosed as hereditary cerebellar ataxia. On the basis of clinical, neuropathological and genetic findings, the authors diagnosed the Gerstmann-Straussler-Scheinker disease, a familial prion disease with an autosomal dominant character. This is the first report on this disease in China. PMID- 17080721 TI - Severe cortical involvement in MV2 Creutzfeldt-Jakob disease: an autopsy case report. AB - MV2 type sporadic Creutzfeldt-Jakob disease (sCJD) is reported to have a long duration and marked involvement of the cerebral deep gray matter. We describe an autopsied long-surviving sCJD case of MV2. In the early stages, the patient exhibited memory impairment, attention deficit and semantic memory disorder. Diffusion-weighted MRI showed abnormal hyperintensity signals along the cerebral cortex, sparing the thalami and basal ganglia. Pathological observations included: severe spongiosis throughout the cerebral cortex, several kuru plaques and plaque-like PrP deposits in the cerebellum, with only minimal degeneration in the thalami and basal ganglia. Our case suggests that MV2 has a wide clinicopathological spectrum, which ranges from "VV2" to "MM2" type. PMID- 17080722 TI - Rapidly enlarging chordoid meningioma with abundant mucin production. AB - This 77-year-old woman with a rapidly enlarging chordoid meningioma first noticed a growing, non-pulsatile, nonpainful soft mass in the left temporal region after a head trauma 2 years earlier. Neuroimaging showed a homogeneously enhanced osteolytic mass lesion in the left temporal bone. Surgery revealed an extradural tumor without significant adhesions. Histopathologically it was a meningioma with features reminiscent of chordoma. Most of the tumor cells contained mucin-rich chordoid elements. Immunohistochemically, the lesion was positive for vimentin and epithelial membranous antigen; it was negative for cytokeratin and S-100 protein. Based on these findings a diagnosis of chordoid meningioma was made. We posit that the rapid enlargement of the tumor over a relatively short period was attributable to its high mucin-producing activity. PMID- 17080723 TI - BCR gene disruption in a pilomyxoid astrocytoma. AB - We report here a 4-month-old child with a large, solid enhancing mass involving predominantly the suprasellar and diencephalic regions, with extension of both hemispheres. The patient underwent partial resection of the mass by right temporal craniotomy. Histological diagnosis was of a low-grade glioma consistent with pilomyxoid astrocytoma. Cytogenetic analyses revealed an insertion on chromosome 17 that involved disruption of the BCR gene. This finding suggests a possible rearrangement of this gene that could act in a similar way to chronic myeloid leukemia with formation of a chimeric tyrosine kinase protein. This study may suggest the use of inhibitors of tyrosine kinase proteins as an alternative treatment approach in cases of refractory or disseminated pilocytic astrocytomas. PMID- 17080724 TI - An autopsy case of frontotemporal dementia with severe dysarthria and motor neuron disease showing numerous basophilic inclusions. AB - We report a clinicopathological study of a patient suffering from frontotemporal dementia (FLD) with severe dysarthria and concomitant motor neuron disease (MND). The patient was a 52-year-old woman with almost simultaneous emergence of severe dysarthria and FTD. The severe dysarthria subsequently evolved into anterior opercular syndrome. Motor neuron signs then emerged, and the patient developed akinetic mutism approximately 2 years after the onset of the disease. The patient died of pneumonia after a 7-year clinical illness. Pathologically, severe and widespread degeneration in the frontal and temporal lobes, including the anterior opercular area, limbic system, basal ganglia, spinal cord and cerebellum, and frequent ubiquitin- and tau-negative basophilic inclusions were observed. The pyramidal tracts and anterior horns of the cervical cord also showed marked degeneration. Cases showing basophilic inclusions reported so far have been divided into two groups: early onset FTD and MND with basophilic inclusions. Our case presented clinicopathological features of both FTD and MND, which suggests that cases showing basophilic inclusions may constitute a clinicopathological entity of FTD/MND. PMID- 17080725 TI - Tauopathy: an overview. AB - In this symposium, cutting-edge data on neuropathology, pathological biochemistry and molecular biology of sporadic and familial tauopathies, as well as on the development of model animals, were presented and discussed by five expert neuropathologists. PMID- 17080726 TI - Cellular tau pathology and immunohistochemical study of tau isoforms in sporadic tauopathies. AB - Pathological inclusions in neurons and glial cells containing fibrillary aggregates of abnormally hyperphosphorylated tau protein are characteristic features in sporadic tauopathies. In the first part of this paper we outline the morphological features of some major sporadic tauopathies. In the second part, to better define the tau isoform composition, we report on the immunohistochemistry of tau isoforms in autopsied brains, including two cases with AD, two with diffuse neurofibrillary tangles with calcification, four with Pick's disease with Pick bodies (PiD), seven with progressive supranuclear palsy (PSP), six with corticobasal degeneration (CBD) and seven cases with argyrophilic grain disease. We used two monoclonal antibodies, RD3 and RD4, and a polyclonal antibody for exon 10 that effectively distinguish between three-repeat (3R) tau and four repeat (4R) tau. Neuronal neurofibrillary tangles (NFT) in AD and diffuse neurofibrillary tangles with calcification contained both 3R-tau and 4R-tau. The Pick bodies were immunopositive for 3R-tau in two cases; however, in two other cases they were mainly immunopositive for 4R-tau. Thus, Pick bodies demonstrated heterogeneity. 3R-tau PiD contained 3R-tau glial inclusions, and 4R-tau PiD contained mainly 4R-tau glial inclusions. Glial inclusions were more abundant in 4R-tau PiD cases. In progressive supranuclear palsy and CBD, both neuronal and glial tau accumulation forming NFF, pretangles, tuft-shaped astrocytes, astrocytic plaques, coiled bodies and threads demonstrated 4R-tau in the cerebral cortices, although in the basal ganglia and brainstem neuronal and glial inclusions were occasionally immunopositive for 3R-tau in addition to 4R-tau. Argyrophilic grains (AG) were immunopositive for 4R-tau, although pretangles were weakly stained for 4R-tau. Thus the immunoreactivity for 4R-tau was different between AG and pretangles. Therefore, the isoform composition on immunohistochemical study showed heterogeneity in PiD, and was not uniform in the basal ganglia and brain stem in PSP and CBD. It is suggested that the isoform composition of sporadic tauopathies may have a spectrum in individual cases, and cellular isoform composition may differ in various brain regions. PMID- 17080727 TI - Neuropathology of familial tauopathy. AB - Frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17) is a hereditary progressive neurodegenerative disorder. FTDP-17 was originally defined in Ann Arbor, Michigan, in 1996. Since then, more than 100 families with FTDP-17 have been described throughout the world, including 18 families identified in Japan. Genetic studies have identified 40 different mutations in the microtubule associated protein tau (MAPT) gene. The clinical features of FTDP-17 are characterized by behavioral, cognitive and motor disturbances that may occur in various combinations and degrees. Neuropathologic examination shows that various degrees of atrophy may be present in the frontal and temporal lobes, basal ganglia, amygdala and hippocampus. All the brains from patients with FTDP-17 have also shown the presence of tau deposits in neurons and glial cells. Mutations in MAPT may result in the increased splicing of exon 10, leading to 4-repeat tau depositions in both neurons and glial cells. MAPT mutations outside of exon 10 show 3- and 4-repeat tau deposits, predominantly in neurons with less glial pathology. Neuronal pathology may resemble that of Alzheimer's disease or Pick's disease because of the presence of neurofibrillary tangles or Pick-like bodies, whereas glial pathology may resemble that of progressive supranuclear palsy or corticobasal degeneration because of the presence of coiled bodies, tufted astrocytes or astrocytic plaques. Correlations between genetic mutations and the heterogeneity of clinical and neuropathologic features remain unclear. PMID- 17080728 TI - Ultrastructural characteristics of tau filaments in tauopathies: immuno-electron microscopic demonstration of tau filaments in tauopathies. AB - The microtubule-associated protein tau aggregates into filaments in the form of neurofibrillary tangles, neuropil threads and argyrophilic grains in neurons, in the form of variable astrocytic tangles in astrocytes and in the form of coiled bodies and argyrophilic threads in oligodendrocytes. These tau filaments may be classified into two types, straight filaments or tubules with 9-18 nm diameters and "twisted ribbons" composed of two parallel aligned components. In the same disease, the fine structure of tau filaments in glial cells roughly resembles that in neurons. In sporadic tauopathies, individual tau filaments show characteristic sizes, shapes and arrangements, and therefore contribute to neuropathologic differential diagnosis. In frontotemporal dementias caused by tau gene mutations, variable filamentous profiles were observed in association with mutation sites and insoluble tau isoforms, including straight filaments or tubules, paired helical filament-like filaments, and twisted ribbons. Pre embedding immunoelectron microscopic studies were carried out using anti-3-repeat tau and anti-4-repeat tau specific antibodies, RD3 and RD4. Straight tubules in neuronal and astrocytic Pick bodies were immunolabeled by the anti-3-repeat tau antibody. The anti-4-repeat tau antibody recognized abnormal tubules comprising neurofibrillary tangles, coiled bodies and argyrophilic threads in progressive supranuclear palsy (PSP) and corticobasal degeneration. In the pre-embedding immunoelectron microscopic study using the phosphorylated tau AT8 antibody, tuft shaped astrocytes of PSP were found to be composed of bundles of abnormal tubules in processes and perikarya of protoplasmic astrocytes. In this study, the 3 repeat tau or 4-repeat tau epitope was detected in situ at the ultrastructural level in abnormal tubules in representative pathological lesions in Pick's disease, PSP and corticobasal degeneration. PMID- 17080729 TI - Biochemistry and molecular biology of tauopathies. AB - Filamentous tau deposits in neurons or glial cells are the hallmark lesions of neurodegenerative tauopathies, such as Alzheimer's disease, Pick's disease, corticobasal degeneration and progressive supranuclear palsy. Biochemical analyses of Sarkosyl-insoluble tau from brains with tauopathies have revealed that tau deposits in different diseases consisted of different tau isoforms (i.e., all six tau isoforms occur in Alzheimer's disease, four repeat tau isoforms occur in corticobasal degeneration or progressive supranuclear palsy, and three repeat tau isoforms occur in Pick's disease). The discovery of mutations in the tau gene in FTDP-17 has established that abnormalities in tau function or expression are sufficient to cause filamentous aggregation of hyperphosphorylated tau and neurodegeneration similar to that seen in sporadic tauopathies. Because the number of tau inclusions and their regional distribution correlate with clinical symptoms, inhibition of tau aggregation or filament formation in neurons or glial cells may prevent neurodegeneration. We have investigated the effects of 42 compounds belonging to nine different chemical classes on tau filament formation, and found that several phenothiazine and polyphenol compounds, and one porphyrin compound inhibit tau filament formation. PMID- 17080730 TI - Animal models of tauopathies. AB - Intracellular fibrillar amyloid lesions comprised of tau proteins are pathological hallmarks in diverse neurodegenerative disorders. As models of these tauopathies, transgenic mice overexpressing tau with or without mutations discovered in familial tauopathies were generated. Findings in these tau transgenic mice support the notion that impairments of tau proteins are causally related to tauopathies, while studies on crossbred mice have indicated initiation and promotion of tau-positive neuropathologies by crosstalk among several pathogenic molecules. Enhancement of tau pathology by amyloid beta (Abeta) deposition provided some of the most compelling evidence for such a cross-talk, and molecular processes linking abnormalities of Abeta and tan have been suggested to involve activation of calcium-dependent protease, calpain, based on analyses of amyloid precursor protein transgenic mice crossbred with other genetically engineered mice with altered calpain activity. It also should be noted that mice transgenic for both tau and alpha-synuclein exhibit facilitated polymerization of these molecules into pathological filaments. Roles of fibrillar tau deposits in nervous system injuries can be mechanistically pursued by longitudinal monitoring of brain amyloidosis and neuroglial degeneration in the time course of antiamyloid intervention. The possibility of in vivo detection of tau-positive amyloid lesions has been demonstrated by intravenous administration of potential tracers into tau transgenic mice and subsequent brain imaging. Moreover, visualization of glial responses in living brains may allow sensitive detection of degenerative changes in the central nervous system. PMID- 17080731 TI - An autopsied case of a 10-month-old baby with hepatosplenomegaly, high fever and necrotizing encephalopathy. PMID- 17080732 TI - Contrasts, conflicts, and change: a case in cultural oncology revisited. PMID- 17080733 TI - Hypovolemia and dehydration in the oncology patient. AB - Dehydration is commonly but often inappropriately diagnosed in cancer patients. Dehydration is the loss of water from the intracellular compartment due to hypernatremia. Dehydration can occur among patients who are hypervolemic, euvolemic, or hypovolemic. Cancer patients are more often hypovolemic, reflecting depletion of water from the extracellular space due to excessive loss, such as from vomiting and diarrhea, or inadequate intake of fluids. Hypovolemia can be hypernatremic, eunatremic, or hyponatremic. The appropriate state of the patient should be determined prior to attempts at correcting the problem. A hyponatremic patient would rehydrate more quickly with a solution higher in sodium, whereas this solution could be dangerous for a hypernatremic patient. Rapid or inappropriate treatment of hypernatremia can lead to death. Subjective findings, physical findings, and laboratory values will help direct the appropriate resuscitation methods. This paper reviews the physiologic control of extracellular volume and electrolytes, diagnosis of sodium and water balance problems, and the management of these concerns. PMID- 17080734 TI - Endoscopic ultrasound-guided celiac plexus neurolysis for pancreatic cancer pain: a single-institution experience and review of the literature. AB - Pancreatic cancer is a common gastrointestinal malignancy with a poor prognosis. The primary goal for caregivers is effective palliative care, especially pain control, which is routinely managed by administration of narcotic analgesics. An alternative or adjunctive modality is celiac plexus neurolysis (CPN), a safe and effective procedure. Recent advances in the use of endoscopic ultrasonography (EUS) have made it an attractive guidance technique for CPN while allowing for a simultaneous tissue diagnosis. We report our experience using EUS-guided CPN and review the available literature regarding this modality. PMID- 17080735 TI - Infusion of palonosetron plus dexamethasone for the prevention of chemotherapy induced nausea and vomiting. AB - Serotonin (5-HT3) receptor antagonists are the foundation of standard antiemetic care for cancer patients receiving emetogenic chemotherapy. To enhance the efficacy of these supportive care agents, dexamethasone is routinely admixed with the 5-HT3 receptor antagonist, which is administered by intravenous infusion before chemotherapy begins. This phase II study evaluated the safety and efficacy of intravenous palonosetron admixed with dexamethasone to prevent chemotherapy induced nausea and vomiting (CINV) in patients receiving moderately emetogenic chemotherapy. Cancer patients received palonosetron 0.25 mg plus dexamethasone 8 mg admixed in 50 mL of infusion solution before receiving at least one qualifying chemotherapeutic agent (cyclophosphamide < or = 1,500 mg/m2, doxorubicin > or = 20 mg/m2, carboplatin, or oxaliplatin). Patients used diaries to record nausea and emesis experienced and rescue medications used. Of 32 participants, 27 (84%) had a complete response (no emesis and no rescue medication) during the acute (0 24 hours) interval posttherapy, 19 (59%) had a complete response during the delayed (> 24-120 hours) posttherapeutic interval, and 19 (59%) had a complete response during the overall (0-120 hours) posttreatment interval. A total of 23 patients (72%) had no emetic episodes, 16 (50%) had no nausea, and 21 (66%) used no rescue medication throughout the overall 5-day interval. The combination was well tolerated. Palonosetron plus dexamethasone given as a pretreatment infusion is effective and safe in preventing acute and delayed CINV in patients receiving moderately emetogenic chemotherapy. PMID- 17080736 TI - A prospective investigation of chemotherapy-induced neutropenia and quality of life. AB - This prospective study of chemotherapy-induced neutropenia (CIN) explored the association between the relative grade of neutropenia and symptom burden and quality of life (QOL). Eighty-four adult cancer patients from nine community oncology centers receiving 1 of 13 myelosuppressive chemotherapies were evaluated at days 0, 4, 7, 9, 11, 14, and 21 of their respective first cycle. Neutropenia grade (grade 3/4 vs grades 0 to 2) was determined by serial absolute neutrophil count (ANC) measures. Measures of patient-reported outcomes included the Rotterdam Symptom Checklist (RSCL), Hospital Anxiety and Depression Scale (HADS), Cancer Care Monitor-Medical Isolation Scale (CCM-MIS), and SF-36. Changes in outcomes from baseline to highest grade of neutropenia were evaluated using mixed model-repeated measures for each of 15 outcomes. Compared with grades 0-2, grade 3/4 neutropenia was associated with greater symptom burden and worse QOL for six measures (P < 0.05). The pattern of differences suggested that measures of symptom distress and social functioning were sensitive to patient changes associated with grade 3/4 neutropenia. Worsening of symptom burden and QOL appears to be associated with severe afebrile neutropenia. A causal relationship between neutropenia and worse symptoms and QOL remains difficult to establish due to the confounding of the effects of neutropenia with other adverse effects of chemotherapy. PMID- 17080737 TI - How to manage steroid diabetes in the patient with cancer. PMID- 17080738 TI - Second Annual Chicago Supportive Oncology Conference: treating symptoms, side effects, and the state of illness. PMID- 17080739 TI - [The surgical treatment of thoracic outlet syndrome]. AB - In this paper the surgical treatment of thoracic outlet syndrome (TOS) is presented. MATERIAL AND METHODS: The investigation of 74 patients treated for vascular complications of the TOS the Paget-Schroetter syndrome and postthrombotic syndrome (27 patients), as well as upper limb ischemia and/or the subclavian artery aneurysm (14 patients) and for neurological complications of the TOS (33 patients) was performed. In case of venous thrombosis the treatment consisted of thrombolytic therapy and decompressive procedures that included transaxillary first rib resection and other surgical procedures that excise different anomalies in the region of the thoracic outlet. In case of complications associated with compression of the subclavian artery the operation consisted of resection of the first rib and accessory osseous and muscular pathological elements using supraclavicular and/or infraclavicular approach with implantation of vascular bypass were performed. In the neurological syndrome of TOS the operation consisted of transaxillary first rib resection and other surgical procedures that excise different anomalies in the region of the thoracic outlet. The therapy results were estimated with use of the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire of the American Academy of Orthopedic Surgeons. RESULTS: In case of venous complications the thrombolytic therapy was successful in all cases the restoration of complete patency was obtained. In all patients first rib was resected via transaxillary approach. The DASH questionnaire revealed the full return of upper limb function in most of patients. Improvement was obtained the in the cases with postthrombotic syndrome. In case of upper limb ischemia the improvement of blood flow was obtained after the subclavian-brachial bypass implantation (5 patients). In this group the DASH score showed return to full activity. In case of subclavian artery aneurysm surgery (9 patients) the DASH scale revealed worsening of limb function in 4 patients during the follow-up period. In the neurological syndrome of TOS the restoration of complete activity of limb was observed. The DASH scale revealed worsening of limb function in 4 patients during the follow-up period. CONCLUSIONS: The optimal therapy of vascular complications is multimodal treatment (thrombolysis or reconstructive vascular procedure with decompressive surgery). The surgical treatment of neurological TOS halts degradation of brachial splice. The decompression of neurovascular bundle in vascular TOS should include the first rib resection in each case. PMID- 17080740 TI - [Subclinical hypothyroidism influences ventricular repolarization measured by QTc interval]. AB - Thyroid disorders exert a major effect on cardiac function and on ECG. Few studies examined the effect of thyroid hormones on ventricular repolarization, measured by corrected QT interval (QTc). Prolonged QTc is associated with increased risk of arrythmias and cardiac mortality. The aim of this study was to examine the influence of subclinical hypothyroidism on ventricular repolarization measured by corrected QTc in standard 12-lead electrocardiogram. The examined group consisted of 32 patients with hypothyroidism, the controls were 39 healthy individuals. The mean corrected QTc was 0.434, SD +/- 0.0296 seconds and 0.414, SD +/- 0.0208 in the examined groups and in controls, respectively (p < 0.01). QTc did not correlate with free thyroxine concentrations (p = 0.4064). In the group with subclinical hypothyroidism we did not observe a significant difference in heart rate (74.3 vs. 73.7, SD +/- 13.58, NS). CONCLUSION: Corrected QT intervals were significantly prolonged in patients with subclinical hypothyroidism. PMID- 17080741 TI - [Quality of life predictors among patients suffering from systemic lupus erythematosus]. AB - Researches on quality of life have been undertaken from dozens of years. However, there are few information about quality of life among people suffering from systemic lupus erythematosus (SLE). The aim of this study was to set predictors making possible quality of life anticipation. The participants of the studied group were 42 patients with inactive illness stage (disease severity were estimated by SLAM scale). Quality of life was measured by own questionnaire. Personal resources, which are the base of anticipation, were estimated by standardized tests (AIS, LOT, GSES). The results indicated that two of examined variables became the quality of predictors: helpless/hopeless strategy and illness acceptation. PMID- 17080742 TI - [Thrombo-embolic events in patients with mechanical prosthetic valves- echocardiography in diagnostic and therapeutic decision making]. AB - Mechanical prosthetic valves (PV) are prone to thrombosis which may result in death or cripple hood due to PV blockade and embolisation (EM). In most pts with clinical symptoms of PV thrombosis (TPV) and in pts with a history of EM transthoracic echocardiography (TTE) and Doppler echocardiography are sufficient in diagnosing but not in stating the exact mechanism of PV stenosis. Transesophageal echocardiography (TEE) is a method of choice in the therapeutic decision making in this group of pts. PV replacement is indicated in pts with a thrombus floating in the left atrium (LA), large, peduncle-like thrombus in LA, TPV in mitral or aortic position with blockade of the disc and symptoms of heart failure < III NYHA, TPV with a high risk of spontaneous or post-thrombolytic EM (large thrombus on the PV annulus >5 mm or >0.8 cm2, highly mobile thrombus, history of EM). Thrombolysis is recommended in tricuspid TPV, TPV in mitral or aortic position with blockade of the disc and symptoms of heart failure III/IV NYHA. Intravenous heparin infusion can be effective as a short-term bridging to PV replacement or an alternative for thrombolysis in case of TPV with no signs of disc blockade. The results of thrombolysis and heparin infusion should be controlled by TEE. In pts with proper function of PV on clinical examination and negative history of EM, TTE/TEE should be considered if the risk factors of EM are present: incorrect INR control, atrial fibrillation, left ventricular dysfunction, III/IV NYHA, diabetes, certain types of PV cage PV, pivoting-disc PV other than Medtronic-Hall valve. PMID- 17080743 TI - [Advanced glycation end products (AGEs) and renal failure]. AB - In the last decade a central role has been attributed to advanced glycation end products (AGE's) in the progression of accelerated atherosclerosis and progresion of chronic renal failure, especially in diabetic nephropathy. Deleterious effects of high AGE's concentration was observed mainly in diabetic hemodialyzed patients. D. M. type 2 is the most common cause of renal failure and renal replacement therapy in Poland and in the world. Diabetic complications are of huge medical and social interest. Therefore it is import ant to reveal all the mechanisms responsible for development of diabetic complications especially diabetic nephropatyhy. A significant role is also played by high cardiovascular mortality in hemodialysis patients, which is caused by atherosclerotic changes in vessels. PMID- 17080744 TI - [Clinical and metabolic consequences of uremic toxicity]. AB - Retention of many substances takes place in the pathogenesis of uremic toxicity. There are almost 100 different molecules described and defined as uremic toxins. These substances are divided into three groups according to EUTOX group calssification. Small water soluble molecules with a molecular weight less than 500 D are included into the first group. Derivate of guanidines, purines, pyrimidines and methyloamines appeared in this group. There is also an unclassified subgroup with urea as a "classical" toxin which the real role in the uraemic syndrome is still discussed. Main symptoms caused by these molecules are digestive disturbances, neurological changes, hypertension etc. We can eliminate almost all of these toxins with standard methods used during dialysotherapy. Substances with a different molecular weight but connected with proteins determine the second group. AGE-s, phenol derivates, leptin and poliamines beside others create this group. There are many studies that have proved that these toxins cause hypertension, arteriosclerosis and shortened life time of hemodialysed patients. However, melatonin toxicity is not fully proved. Different types of renal replacement therapy are not valid to purify blood from protein bound substances. Middle molecules are included into the third group, with a molecular weight higher than 500 D. There are cytokines, neuro-transmitters e.g. beta-endorphin, metencephalin and many others accounted into this group. One of them is the parathormon, well known and considered as "universal" toxin for several years. Middle molecules are causing very different effects. They are responsible for: anemia, arteriosclerosis, chronic inflammation and generally increase dialysed patient mortality. Toxic action of several molecules described below is still not proved; however there are some ongoing studies aimed to find pathophysiological links between old and new described uremic toxins. PMID- 17080745 TI - [Fabry disease]. AB - Fabry disease is a rare genetic disorder, which is linked to a defect of alfa galactosidase. In consequence it leads to an excess of glicosphyngolipids in lysosomes of various tissues and organs. Clinical symptoms are related to heart, skin, kidneys and nervous system. Nowadays due to a possibility of substitution of galactosidase A, a influence on clinical course of the disease can be attained: arresting of progression and avoidance of complications. PMID- 17080746 TI - [Skin manifestations of Lyme borreliosis]. AB - Lyme borreliosis is the most common tick-borne disease which is caused by Borrelia burgdorferi and transmitted in the Poland, as well as Europe, primarily by Ixodes ticks. After inoculation spirochetes spreads in the skin, activate locally immune response mechanisms and cause characteristic skin lesions like erythema migrans, lymphadenosis benigna cutis, erythema migrans multiplex and acrodermatitis chronica atrophicans. The correct clinical diagnosis of skin lesions and antibiotic treatment is most important for regression of symptoms and prevention of late manifestations of Lyme disease. PMID- 17080747 TI - [Outbreak of Lyme borreliosis with joint manifestation--case report]. AB - Lyme borreliosis is caused by the spirochete Borrelia burgdorferi, which is transmitted into the human body by ticks. The clinical symptoms are associated with skin, joints, heart and nervous system. Four groups of antibiotics are used in Lyme borreliosis treatment: penicillins, cephalosporins, tetracyclines and macrolides. We present the case of Lyme borreliosis with outbreak joint manifestation. PMID- 17080748 TI - [Early metabolic abnormalities--insulin resistance, hyperinsulinemia, impaired glucose tolerance and diabetes, in adolescent girls with polycystic ovarian syndrome]. AB - Polycystic ovary syndrome (PCOS), the main androgen disorder in women. Recently, it has been suggested that the condition is hereditary and that women with PCOS have disturbances, such as hyperandrogenism and additional metabolic abnormalities as hyperinsulinaemia, increased insulin resistance, dyslipidemia. Polycystic ovary syndrome (PCOS) usually arises during puberty and is marked by hyperinsulinemia and hyperandrogenism. Adolescents with PCOS are at an increased risk of developing health problems later on in life such as type 2 diabetes, cardiovascular disease, and infertility. Furthermore, the physical signs of PCOS can be detrimental to a teenage girl's self-image. Early diagnosis and treatment of PCOS in adolescents are essential in ensuring adulthood health and restoring self-esteem. Treatment for an adolescent with PCOS includes diet and exercise, metformin, and oral contraceptive pills. Still, surgery is not indicated in teenagers. Furthermore, psychological factors must be taken into consideration as well. PMID- 17080749 TI - [Heterotopic pregnancy after in vitro fertilization]. AB - In our paper we described a rare case of heterotopic pregnancy after in vitro fertilization (IVF-ET). Heterotopic pregnancy (coexistence of intrauterine and ectopic pregnancy) appears in 1 per 30 thousand cases of natural pregnancies and in 1% of pregnancies after IVF. The pregnancy was obtained after ICSI and transfer of 2 embryos. The pregnancy was confirmed by HCG serum presence--13 days after ET. On day 39 after IVF procedure, the patient was admitted to the hospital because of abdominal pain. At admission, ultrasound examination confirmed an 8 week intrauterine pregnancy and enlarged polycystic ovaries. After 3-day-long observation, patient status worsened and laparoscopy procedure was performed. The ectopic pregnancy was confirmed during operation and the ovum was removed a long with fallopian tube. Ultrasound examination on the 6th day after operation confirmed single intrauterine viable fetus. in order to diagnose the possible complications, women undergoing IVF procedure should be carefully observed by IVF performing physicians. PMID- 17080750 TI - Power and sample size considerations in clinical trials with competing risk endpoints. AB - In clinical trials with a time-to-event endpoint, subjects are often at risk for events other than the one of interest. When the occurrence of one type of event precludes observation of any later events or alters the probably of subsequent events, the situation is one of competing risks. During the planning stage of a clinical trial with competing risks, it is important to take all possible events into account. This paper gives expressions for the power and sample size for competing risks based on a flexible parametric Weibull model. Nonuniform accrual to the study is considered and an allocation ratio other than one may be used. Results are also provided for the case where two or more of the competing risks are of primary interest. PMID- 17080751 TI - Surrogate threshold effect: an alternative measure for meta-analytic surrogate endpoint validation. AB - In many therapeutic areas, the identification and validation of surrogate endpoints is of prime interest to reduce the duration and/or size of clinical trials. Buyse et al. [Biostatistics 2000; 1:49-67] proposed a meta-analytic approach to the validation. In this approach, the validity of a surrogate is quantified by the coefficient of determination Rtrial2 obtained from a model, which allows for prediction of the treatment effect on the endpoint of interest ('true' endpoint) from the effect on the surrogate. One problem related to the use of Rtial2 is the difficulty in interpreting its value. To address this difficulty, in this paper we introduce a new concept, the so-called surrogate threshold effect (STE), defined as the minimum treatment effect on the surrogate necessary to predict a non-zero effect on the true endpoint. One of its interesting features, apart from providing information relevant to the practical use of a surrogate endpoint, is its natural interpretation from a clinical point of view. PMID- 17080752 TI - Identifying the most successful dose (MSD) in dose-finding studies in cancer. AB - For a dose finding study in cancer, the most successful dose (MSD), among a group of available doses, is that dose at which the overall success rate is the highest. This rate is the product of the rate of seeing non-toxicities together with the rate of tumor response. A successful dose finding trial in this context is one where we manage to identify the MSD in an efficient manner. In practice we may also need to consider algorithms for identifying the MSD which can incorporate certain restrictions, the most common restriction maintaining the estimated toxicity rate alone below some maximum rate. In this case the MSD may correspond to a different level than that for the unconstrained MSD and, in providing a final recommendation, it is important to underline that it is subject to the given constraint. We work with the approach described in O'Quigley et al. [Biometrics 2001; 57(4):1018-1029]. The focus of that work was dose finding in HIV where both information on toxicity and efficacy were almost immediately available. Recent cancer studies are beginning to fall under this same heading where, as before, toxicity can be quickly evaluated and, in addition, we can rely on biological markers or other measures of tumor response. Mindful of the particular context of cancer, our purpose here is to consider the methodology developed by O'Quigley et al. and its practical implementation. We also carry out a study on the doubly under-parameterized model, developed by O'Quigley et al. but not PMID- 17080753 TI - Comparison of models for average bioequivalence in replicated crossover designs. AB - Average bioequivalence (ABE) has been the regulatory standard for bioequivalence (BE) since the 1990s. BE studies are commonly two-period crossovers, but may also use replicated designs. The replicated crossover will provide greater power for the ABE assessment. FDA has recommended that ABE analysis of replicated crossovers use a model which includes terms for separate within- and between subject components for each formulation and which allows for a subject x formulation interaction component. Our simulation study compares the performance of four alternative mixed effects models: the FDA model, a three variance component model proposed by Ekbohm and Melander (EM), a random intercepts and slopes model (RIS) proposed by Patterson and Jones, and a simple model that contains only two variance components. The simple model fails (when not 'true') to provide adequate coverage and it accepts the hypothesis of equivalence too often. FDA and EM models are frequently indistinguishable and often provide the best performance with respect to coverage and probability of concluding BE. The RIS model concludes equivalence too often when both the within- and between subject variance components differ between formulations. The FDA analysis model is recommended because it provides the most detail regarding components of variability and has a slight advantage over the EM model in confidence interval length. PMID- 17080754 TI - Comparing proportional hazards and accelerated failure time models: an application in influenza. AB - The proportional hazards (PH) model is routinely employed for the analysis of time-to-event data in medical research when it is required to assess the effect of an intervention in the presence of covariates. The assumption of PH required for the PH approach may not hold, especially in circumstances where the effect of the intervention is to delay or accelerate the onset of an event rather than to reduce or increase the overall proportion of subjects who observe the event through time. If the assumption of PH is violated, the results from a PH model will be difficult to generalize to situations where the length of follow-up is different to that used in the analysis. It is also difficult to translate the results into the effect upon the expected median duration of illness for a patient in a clinical setting. The accelerated failure time (AFT) approach is an alternative strategy for the analysis of time-to-event data and can be suitable even when hazards are not proportional and this family of models contains a certain form of PH as a special case. The framework can allow for different forms of the hazard function and may provide a closer description of the data in certain circumstances. In addition, the results of the AFT model may be easier to interpret and more relevant to clinicians, as they can be directly translated into expected reduction or prolongation of the median time to event, unlike the hazard ratio. We recommend that consideration is given to an AFT modelling approach in the analysis of time-to-event data in medical research. PMID- 17080755 TI - Teaching statistics to clinical research staff in a pharmaceutical company. AB - Education of clinical research staff in understanding statistical concepts is an area of importance for pharmaceutical companies. This understanding is needed to help them communicate with statisticians using a common language, in designing clinical trials and interpretation of clinical trial results. Such staff has little time for a one-semester or even a one-week continuing education course in statistics. Faced with this reality, we developed a 3-module course,for a total of 1.5 days, which was taught over a period of one month that addresses the needs of this audience. We describe the format and content of the course and provide references that can serve as a resource for teaching such a course. PMID- 17080756 TI - Comment on 'Comparison of sample size formulae for 2 x 2 cross-over designs applied to bioequivalence studies', Siqueira AL, Whitehead A, Todd S and Lucini MM. Pharmaceutical Statistics 2005; 4:233-243. PMID- 17080757 TI - [Farewell to a pioneer in the campaign against tuberculosis. Prof. Dr. med. Eduard Haeflinger]. PMID- 17080758 TI - [CME ultrasound diagnosis 13. Right-sided hip pain]. PMID- 17080759 TI - [Tuberculosis: a perspective on its development from an epidemic to an endemic disease--reminiscences of a contemporary witness]. AB - The prevalence of infection, i.e. endemic infection with Mycobacterium tuberculosis, which was 100% in post-adolescent adults in 1900, had decreased to 23.1% by the year 1998. In Switzerland, on average 1100 active tuberculosis cases were diagnosed in hospitals (acute-care clinics) between 1990 and 1993. A mass X ray fluoroscopic screening performed in 1943/44 by the Swiss Army detected 967 cases of active tuberculosis, corresponding to 1.9 per thousand of the fluoroscopically screened subjects. In 2000, photofluorography examinations performed on asylum seekers detected 3.25 per thousand cases of active tuberculosis. Since the beginning of 2006, these examinations have no longer been performed to save costs. Back in 1890, 6639 of the patients died of tuberculosis; by 2001, this number had dropped to an incredibly low 22. Our present aim must now be to prevent the primary infections that will later lead to tuberculosis in 5-8% of the cases. Consequently, we will not be able to eradicate tuberculosis in Switzerland for two to three more decades; a feat that might not even be accomplished within in a century in the developing countries. PMID- 17080760 TI - [Pelvic congestion syndrom]. AB - We describe imaging findings and interventional therapy of a 37-year old female patient suffering from chronic pelvic pain with associated vulvar varicosis. MR angiography showed multiple varices in the left hemi-pelvis, a dilated draining vein from the left ovary as well as a left-sided vulvar varicosis, which was clinically evident. In addition, the MRI suggested insufficiency of the great saphenous vein with varicosis of its tributaries. After crossectomy and infragenual stripping of the great saphenous vein, we proceeded with a left phlebectomy and ligation of the supplying veins. Due to post-operative persistence of the vulvar varicosis the patient underwent coil-embolisation and liquid sclerotherapy (Aethoxysklerol) of the varicose pelvic veins and the left ovarian vein with good results. PMID- 17080761 TI - [Blue discoloration of hands, numbness of feet, indolent cervical lymph node swelling in a 73-year-old man]. AB - A 73-years old patient came to our outpatient clinic because of a blue discoloration of his hands. Furthermore there was a systemic inflammatory reaction, burning pain and numbness of his feet due to a polyneuropathy, and an indolent enlargement of the cervical lymph nodes. In 1996 and 2001 cervical lymph node resections were done because of localized angiofollicular lymphnode hyperplasia (Castleman's disease). The laboratory values confirmed a systemic inflammatory reaction, a hypothyreosis and a monoclonal gammopathy. A CT-scan showed enlarged cervical, intrathoracic and abdominal lymphnodes and a splenomegaly. So all the criterias for a POEMS syndrome (special form of multiple myeloma) were met with polyneuropathy, organomegaly, endocrinopathy, monoclonal protein and skin changes. The generalized lymphadenopathy corresponded histologically to the prior mentioned Castleman's disease. The patient responded well to systemic glucocorticoid treatment and today he is asymptomatic. PMID- 17080762 TI - [CME laboratory 1/solution. Chronic cannabis use]. PMID- 17080763 TI - ["Our God rightly sends miracles only extremely rarely"--Bernadette Soubirous (1844-1879), Lourdes, and the history of miracle cures]. PMID- 17080764 TI - Adaptive design: a concept whose time has come. PMID- 17080765 TI - Adapting the sample size planning of a phase III trial based on phase II data. AB - Traditionally, in clinical development plan, phase II trials are relatively small and can be expected to result in a large degree of uncertainty in the estimates based on which Phase III trials are planned. Phase II trials are also to explore appropriate primary efficacy endpoint(s) or patient populations. When the biology of the disease and pathogenesis of disease progression are well understood, the phase II and phase III studies may be performed in the same patient population with the same primary endpoint, e.g. efficacy measured by HbA1c in non-insulin dependent diabetes mellitus trials with treatment duration of at least three months. In the disease areas that molecular pathways are not well established or the clinical outcome endpoint may not be observed in a short-term study, e.g. mortality in cancer or AIDS trials, the treatment effect may be postulated through use of intermediate surrogate endpoint in phase II trials. However, in many cases, we generally explore the appropriate clinical endpoint in the phase II trials. An important question is how much of the effect observed in the surrogate endpoint in the phase II study can be translated into the clinical effect in the phase III trial. Another question is how much of the uncertainty remains in phase III trials. In this work, we study the utility of adaptation by design (not by statistical test) in the sense of adapting the phase II information for planning the phase III trials. That is, we investigate the impact of using various phase II effect size estimates on the sample size planning for phase III trials. In general, if the point estimate of the phase II trial is used for planning, it is advisable to size the phase III trial by choosing a smaller alpha level or a higher power level. The adaptation via using the lower limit of the one standard deviation confidence interval from the phase II trial appears to be a reasonable choice since it balances well between the empirical power of the launched trials and the proportion of trials not launched if a threshold lower than the true effect size of phase III trial can be chosen for determining whether the phase III trial is to be launched. PMID- 17080766 TI - Methodological issues with adaptation of clinical trial design. AB - Adaptation of clinical trial design generates many issues that have not been resolved for practical applications, though statistical methodology has advanced greatly. This paper focuses on some methodological issues. In one type of adaptation such as sample size re-estimation, only the postulated value of a parameter for planning the trial size may be altered. In another type, the originally intended hypothesis for testing may be modified using the internal data accumulated at an interim time of the trial, such as changing the primary endpoint and dropping a treatment arm. For sample size re-estimation, we make a contrast between an adaptive test weighting the two-stage test statistics with the statistical information given by the original design and the original sample mean test with a properly corrected critical value. We point out the difficulty in planning a confirmatory trial based on the crude information generated by exploratory trials. In regards to selecting a primary endpoint, we argue that the selection process that allows switching from one endpoint to the other with the internal data of the trial is not very likely to gain a power advantage over the simple process of selecting one from the two endpoints by testing them with an equal split of alpha (Bonferroni adjustment). For dropping a treatment arm, distributing the remaining sample size of the discontinued arm to other treatment arms can substantially improve the statistical power of identifying a superior treatment arm in the design. A common difficult methodological issue is that of how to select an adaptation rule in the trial planning stage. Pre-specification of the adaptation rule is important for the practicality consideration. Changing the originally intended hypothesis for testing with the internal data generates great concerns to clinical trial researchers. PMID- 17080767 TI - Application of an adaptive design to a randomized phase II selection trial in gastric cancer: a report of the study design. AB - Randomized phase II selection trials seek to provide unbiased comparisons for the selection of the most promising treatment arm for evaluation in a future phase III trial. In this paper, we present an application of an adaptive design to a randomized phase II selection trial comparing three experimental treatments with a control arm in patients with advanced gastric cancer. The trial design continuously monitors multiple patient outcomes to protect future patients from treatments with unacceptably high toxicity and/or unacceptably low efficacy. We use a Bayesian approach to monitor the trial and carry out simulations to investigate operating characteristics of the trial design. The simulation study also evaluates the sensitivity of the design to the prior distribution by considering two alternative priors. PMID- 17080768 TI - Operational challenges in adaptive design implementation. AB - Implementation of adaptive clinical trial designs raises challenges with regard to the processes by which accruing trial data is analyzed, reviewed, and acted upon. In line with current monitoring conventions, it should be viewed that inappropriate knowledge of interim results can raise concerns regarding maintaining trial integrity and interpretability of results. Here we discuss issues related to these processes in adaptive trials, and point out distinctions versus other more familiar monitoring situations. One topic involves the composition of the group of individuals who will have access to interim results in order to recommend adaptations. We discuss operational models for data review by this group; one question addressed is whether in adaptive trials a role in this process for a representative of the study sponsor could at times be warranted, and might be justified if adequate protections are in place. Another issue involves whether adaptations made based upon interim data can convey to observers an amount of information about the results, which could rise to a level of concern. We consider whether different types of adaptations might be more or less problematic with regard to this issue, and recommend steps that might be considered to mitigate this concern. PMID- 17080769 TI - Bayesian decision procedures for binary and continuous bivariate dose-escalation studies. AB - In this paper, Bayesian decision procedures are developed for dose-escalation studies based on binary measures of undesirable events and continuous measures of therapeutic benefit. The methods generalize earlier approaches where undesirable events and therapeutic benefit are both binary. A logistic regression model is used to model the binary responses, while a linear regression model is used to model the continuous responses. Prior distributions for the unknown model parameters are suggested. A gain function is discussed and an optional safety constraint is included. PMID- 17080770 TI - Planning for an adaptive design: a case study in COPD. AB - We discuss the practical and clinical considerations encountered when planning a Phase IIa trial in chronic obstructive pulmonary disease (COPD). Various adaptive strategies for reducing the cost of the trial and the statistical implications of these are explored. Use of the EAST software to evaluate the properties of the study designs with one or more interim analyses for futility, efficacy or either is described. We emphasize the rationale for choosing between alternative designs and the relationship between the clinical and statistical considerations. PMID- 17080771 TI - Developing a tier I CAMHS foundation course: report on a 4-year initiative. AB - Despite a growing recognition of the strategic importance of the UK's Tier 1 in supporting children and young people with mental health needs, detailed descriptions of the developmental hurdles and content of such courses remain obscure. There are a number of initiatives being undertaken nationally to remedy this and the aim of this article is to contribute the authors' experience to what is becoming a burgeoning national endeavour. Particular attention is drawn to the problems of developing volume multiagency trainings in areas of significant social deprivation and mental health morbidity but innovation is needed to balance the competing needs for quality, penetration into Tier 1, pressures on staff time and managers' release constraints. A core syllabus is tentatively suggested, with additional skills components, and a teaching session is explored in detail to illustrate the authors' methodology and approach. PMID- 17080772 TI - Parenting programmes for behavioural problems: where do tertiary units fit in a comprehensive service? AB - Children with behavioural problems are often referred to child mental health services without rationalization of criteria or referral routes. The aims of the present study were to establish the psychosocial characteristics of children with behavioural problems referred to a tertiary Day Resource, and to compare these with children referred for similar problems to a specialist outpatient child mental health service and a family support community service. Demographic data, the Strengths and Difficulties Questionnaire and the Health of the Nations Outcome scales for Children and Adolescents were used. Families attending the Day Resource were found to have more involvement with other agencies, and children showed higher levels of clinical severity and complexity. Most of this difference was accounted for by attentional difficulties, as well as communication and physical health problems. These findings confirmed the hypothesis that families seen at tertiary service level presented with more complex psychosocial needs and frequent child comorbidity, thereby justifying the multimodal treatment package developed at that service level. It is concluded that there is a need for this level of parenting interventions, but only for complex cases that have not responded to community interventions. PMID- 17080773 TI - Family impact of neurodevelopmental late effects in survivors of pediatric cancer: review of research, clinical evidence, and future directions. AB - Children and adolescents who have survived childhood cancer are vulnerable to late effects of their disease and treatment experience, including both physical and psychological sequelae. The neurodevelopmental (ND) sequelae (e.g., difficulties with attention, memory, information processing, and other executive functions) faced by children surviving central nervous system (CNS) cancers or CNS treatments can have a range of effects on their psychological adjustment and quality of life as they reintegrate into school and social settings. Survivors facing such difficulties may be influenced by their family environment, and these struggles may in turn impact the perceived burden of their parents and family system, suggesting a bidirectional framework for understanding the impact of ND late effects within the family. The current study summarizes the following: Findings of family outcomes in children with ND late effects of cancer treatment; evaluation of methodological and measurement issues; the importance of examining bidirectional family-child influences; and implications for future survivorship research and clinical care addressing the role of ND late effects from a systems perspective. PMID- 17080774 TI - Solution-focused brief therapy with families who have a child with intellectual disabilities: A description of the content of initial sessions and the processes. AB - Solution-focused brief therapy (SFBT) is used in a range of child services but little is known about its application to families who have a child with severe or profound intellectual disabilities. This qualitative study examines SFBT with seven such families. It considers the content of sessions and the processes that occur from the practitioner's perspective. Participants' experiences are reported elsewhere (Lloyd & Dallos, submitted). A thematic analysis of seven initial sessions indicated that SFBT highlighted parents' competencies, goals and achievements. The 'miracle question' prompted a change in rapport, discussion about the impossibility of the child becoming 'normal' and a shift away from wishful thinking towards problem solving. In sessions processes emerged of empowerment, integration of the goals into the mothers' life narrative, understanding the child's abilities and clarification of the preferred future. These themes resonated with the literature on effective coping styles for these families. The cases suggest that SFBT can generate a useful parent-professional partnership. However, difficulties were encountered when a child with autism participated in the session. Consideration is given to the 'miracle question' and the inclusion of measures of self-efficacy, coping and the therapeutic relationship. PMID- 17080775 TI - An experience of group work with parents of children and adolescents with gender identity disorder. AB - This article gives an account of an experience of group work with parents and carers who had children or adolescents with gender identity disorder (GID). The history of this intervention within the context of a service for children with gender identity problems is outlined. The limited literature on the subject is reviewed. Group meetings were held monthly for 6 months, facilitated by two therapists (the authors). Selection criteria for group participants, the aims of the group and the methodology for achieving those aims are described. Some information about the group's composition is provided. The structure and content of the group sessions are outlined together with details of some group interactions. Finally, we present the results of an evaluation of the intervention through feedback questionnaires and discuss the value for the children and young people of running such groups. PMID- 17080776 TI - A parent-report Gender Identity Questionnaire for Children: A cross-national, cross-clinic comparative analysis. AB - A one-factor, 14-item parent-report Gender Identity Questionnaire for Children (GIQC) was developed in a sample of 325 clinic-referred children with gender identity problems and 504 controls from Toronto, Canada (Johnson et al., 2004). In this study, we report a cross-national, cross-clinic comparative analysis of the GIQC on gender-referred children (N = 338) from Toronto and gender-referred children (N = 175) from Utrecht, The Netherlands. Across clinics, the results showed both similarities and differences. Gender-referred boys from Utrecht had a significantly higher total score (indicating more cross-gender behavior) than did gender-referred boys from Toronto, but there was no significant difference for girls. In the Toronto sample, the gender-referred girls had a significantly higher total score than the gender-referred boys, but there was no significant sex difference in the Utrecht sample. Across both clinics, gender-referred children who met the complete DSM criteria for gender identity disorder (GID) had a significantly higher cross-gender score than the gender-referred children who were subthreshold for GID (Cohen's d = 1.11). The results of this study provide the first empirical evidence of relative similarity in cross-gender behavior in a sample of gender-referred children from western Europe when compared to North American children. The results also provide some support for cross-clinic consistency in clinician-based diagnosis of GID. PMID- 17080777 TI - An examination of cutting and other methods of DSH among children and adolescents presenting to an outpatient psychiatric clinic in New Zealand. AB - This article explores the issue of cutting among children and adolescents presenting to an outpatient child and adolescent mental health service in New Zealand. Of interest is the extent to which children and adolescents who cut themselves also carry out other forms of deliberate self-harm (DSH). This study found that there was much cross over in the methods of DSH used and that cutters were not a distinguishable group. Therefore, those who present to mental health services with cutting need to be treated as carrying significant risk of fatality. PMID- 17080778 TI - Intensive mental health services for children with serious emotional disturbances through a school-based, community-oriented program. AB - Treating children with severe, early onset emotional and behavioral disturbances remains a daunting task for mental health and education systems. This article describes key principles, features, and outcomes for the school-based Intensive Mental Health Program (IMHP), a program designed specifically to provide comprehensive evidence-based, ecologically sensitive, and individualized services for this difficult-to-treat population. Although the IMHP is a relatively new model, preliminary studies of outcomes and treatment processes give initial evidence that most children improve notably in their role performance, behavior, and emotional adjustment over the course of treatment. With increasing demands for effective, affordable school-based mental health services, the IMHP offers a developing but promising prototype for services geared to meet the unique needs of children with complex disturbances of emotions and behavior. PMID- 17080779 TI - 'Outlook': A psychological service for children with a different appearance. AB - 'Outlook' is a unit within North Bristol Trust which has been developing a psychosocial service for children, young people and adults with a different appearance resulting from congenital abnormalities such as cleft lip and palate and also burns and other forms of trauma. This study presents data collected from 29 children aged between 5 and 16 who received individual therapeutic interventions. The baseline assessment included a semistructured interview schedule, visual analogue scales and the CBCL (Achenbach, 1991) to provide the parents' view of their children's behaviour profile. Measures were repeated post intervention and at 6 months follow-up. The intervention consisted of four sessions of cognitive-behavioural therapy (CBT) including social skills and problem-solving components. Statistical analyses revealed significant outcomes on a range of measures. At 6 months follow-up children reported a reduction in the frequency of teasing and in the degree of distress it caused both in the classroom and in the playground. Parents reported reductions in their children's' somatizing behaviour and levels of anxiety. These outcomes suggest that this approach represents an effective therapy for children with appearance-related psychological problems. PMID- 17080780 TI - The management of ADHD and associated problems in a young person with cleidocranial dysostosis (CCD) and mild intellectual disability. AB - It is increasingly recognized that comorbidity is common in all fields of psychiatry, and furthermore, it is acknowledged that a large number of individuals with genetically determined conditions have associated behavioural phenotypes, and are more susceptible to particular psychiatric and psychological comorbidities than others. It is also recognized that the identification of such phenotypes enables clinicians to be more aware of the potential difficulties an individual may experience, and hence, facilitate early diagnosis, effective management and prevention, appropriate allocation of resources and psychoeducation for the individual and their family. We describe the case report of a girl with cleidocranial dysostosis (CCD), and comorbid intellectual disability and attention deficit hyperactivity disorder (ADHD), and suggest the possible existence of a behavioural phenotype. We also highlight the lack of an evidence base for the management of ADHD within the learning-disability population, and describe successful management utilizing the current evidence base, which exists for those of average intellectual ability. PMID- 17080781 TI - Pervasive refusal syndrome among asylum-seeking children. AB - A number of asylum-seeking children in Sweden have developed a pervasive loss of function associated with profound social withdrawal. The syndrome is called Depressive Devitalization. The aim of this study was to identify possible aetiological factors, outline the similarities between Depressive Devitalization and Pervasive Refusal Syndrome and to explore possible differential diagnoses. The research was based on a literature study. Databases searched included PsychINFO, Medline, Pub med, COCHRANE and PILOTS. Possible aetiological factors identified included: Children having a perfectionist, ambitious and conscientious premorbid personality, psychiatric problems of children and parents, and traumatic events. Symptoms between the two syndromes differed only in pattern of refusal and neurological symptoms. None of the differential diagnoses explored could account for all features. The individual impact of aetiological factors requires further investigation. Children might previously have been diagnosed with a number of differential diagnoses, though none of these accounts for all symptoms seen in the syndromes. Depressive Devitalization and Pervasive Refusal Syndrome are suggested to be subgroups of the same refusal syndrome. PMID- 17080782 TI - Review of Liebman et al.'s 'An integrated treatment program for anorexia nervosa'. AB - This article reviews the paper by Liebman, Minuchin, and Baker (1974) describing the use of a family meal as part of an integrated treatment approach for anorexia nervosa. The ideas laid out in the paper are described and discussed in terms of the understanding of anorexia nervosa at the time as well as placed in a current clinical and theoretical context. A comment is made on whether or not, in this author's opinion, the paper stands 'the test of time'. PMID- 17080783 TI - PKU in the family: working together. PMID- 17080784 TI - The 2005 Visualization Career Award: Arie Kaufman. PMID- 17080785 TI - The 2005 Visualization Technical Achievement Award: Charles Hansen. PMID- 17080786 TI - ASK-GraphView: A large scale graph visualization system. AB - We describe ASK-GraphView, a node-link-based graph visualization system that allows clustering and interactive navigation of large graphs, ranging in size up to 16 million edges. The system uses a scalable architecture and a series of increasingly sophisticated clustering algorithms to construct a hierarchy on an arbitrary, weighted undirected input graph. By lowering the interactivity requirements we can scale to substantially bigger graphs. The user is allowed to navigate this hierarchy in a top down manner by interactively expanding individual clusters. ASK-GraphView also provides facilities for filtering and coloring, annotation and cluster labeling. PMID- 17080787 TI - MatrixExplorer: a dual-representation system to explore social networks. AB - MatrixExplorer is a network visualization system that uses two representations: node-link diagrams and matrices. Its design comes from a list of requirements formalized after several interviews and a participatory design session conducted with social science researchers. Although matrices are commonly used in social networks analysis, very few systems support the matrix-based representations to visualize and analyze networks. MatrixExplorer provides several novel features to support the exploration of social networks with a matrix-based representation, in addition to the standard interactive filtering and clustering functions. It provides tools to reorder (layout) matrices, to annotate and compare findings across different layouts and find consensus among several clusterings. MatrixExplorer also supports Node-link diagram views which are familiar to most users and remain a convenient way to publish or communicate exploration results. Matrix and node-link representations are kept synchronized at all stages of the exploration process. PMID- 17080788 TI - Visual analysis of multivariate state transition graphs. AB - We present a new approach for the visual analysis of state transition graphs. We deal with multivariate graphs where a number of attributes are associated with every node. Our method provides an interactive attribute-based clustering facility. Clustering results in metric, hierarchical and relational data, represented in a single visualization. To visualize hierarchically structured quantitative data, we introduce a novel technique: the bar tree. We combine this with a node-link diagram to visualize the hierarchy and an arc diagram to visualize relational data. Our method enables the user to gain significant insight into large state transition graphs containing tens of thousands of nodes. We illustrate the effectiveness of our approach by applying it to a real-world use case. The graph we consider models the behavior of an industrial wafer stepper and contains 55 043 nodes and 289 443 edges. PMID- 17080789 TI - Balancing systematic and flexible exploration of social networks. AB - Social network analysis (SNA) has emerged as a powerful method for understanding the importance of relationships in networks. However, interactive exploration of networks is currently challenging because: (1) it is difficult to find patterns and comprehend the structure of networks with many nodes and links, and (2) current systems are often a medley of statistical methods and overwhelming visual output which leaves many analysts uncertain about how to explore in an orderly manner. This results in exploration that is largely opportunistic. Our contributions are techniques to help structural analysts understand social networks more effectively. We present SocialAction, a system that uses attribute ranking and coordinated views to help users systematically examine numerous SNA measures. Users can (1) flexibly iterate through visualizations of measures to gain an overview, filter nodes, and find outliers, (2) aggregate networks using link structure, find cohesive subgroups, and focus on communities of interest, and (3) untangle networks by viewing different link types separately, or find patterns across different link types using a matrix overview. For each operation, a stable node layout is maintained in the network visualization so users can make comparisons. SocialAction offers analysts a strategy beyond opportunism, as it provides systematic, yet flexible, techniques for exploring social networks. PMID- 17080790 TI - Multi-scale banking to 45 degrees. AB - In his text Visualizing Data, William Cleveland demonstrates how the aspect ratio of a line chart can affect an analyst's perception of trends in the data. Cleveland proposes an optimization technique for computing the aspect ratio such that the average absolute orientation of line segments in the chart is equal to 45 degrees. This technique, called banking to 45 degrees, is designed to maximize the discriminability of the orientations of the line segments in the chart. In this paper, we revisit this classic result and describe two new extensions. First, we propose alternate optimization criteria designed to further improve the visual perception of line segment orientations. Second, we develop multi-scale banking, a technique that combines spectral analysis with banking to 45 degrees. Our technique automatically identifies trends at various frequency scales and then generates a banked chart for each of these scales. We demonstrate the utility of our techniques in a range of visualization tools and analysis examples. PMID- 17080791 TI - Measuring data abstraction quality in multiresolution visualizations. AB - Data abstraction techniques are widely used in multiresolution visualization systems to reduce visual clutter and facilitate analysis from overview to detail. However, analysts are usually unaware of how well the abstracted data represent the original dataset, which can impact the reliability of results gleaned from the abstractions. In this paper, we define two data abstraction quality measures for computing the degree to which the abstraction conveys the original dataset: the Histogram Difference Measure and the Nearest Neighbor Measure. They have been integrated within XmdvTool, a public-domain multiresolution visualization system for multivariate data analysis that supports sampling as well as clustering to simplify data. Several interactive operations are provided, including adjusting the data abstraction level, changing selected regions, and setting the acceptable data abstraction quality level. Conducting these operations, analysts can select an optimal data abstraction level. Also, analysts can compare different abstraction methods using the measures to see how well relative data density and outliers are maintained, and then select an abstraction method that meets the requirement of their analytic tasks. PMID- 17080792 TI - Enabling automatic clutter reduction in parallel coordinate plots. AB - We have previously shown that random sampling is an effective clutter reduction technique and that a sampling lens can facilitate focus+context viewing of particular regions. This demands an efficient method of estimating the overlap or occlusion of large numbers of intersecting lines in order to automatically adjust the sampling rate within the lens. This paper proposes several ways for measuring occlusion in parallel coordinate plots. An empirical study into the accuracy and efficiency of the occlusion measures show that a probabilistic approach combined with a 'binning' technique is very fast and yet approaches the accuracy of the more expensive 'true' complete measurement. PMID- 17080793 TI - Topographic visualization of prefix propagation in the internet. AB - We propose a new metaphor for the visualization of prefixes propagation in the Internet. Such a metaphor is based on the concept of topographic map and allows to put in evidence the relative importance of the Internet Service Providers (ISPs) involved in the routing of the prefix. Based on the new metaphor we propose an algorithm for computing layouts and experiment with such algorithm on a test suite taken from the real Internet. The paper extends the visualization approach of the BGPlay service, which is an Internet routing monitoring tool widely used by ISP operators. PMID- 17080794 TI - Network visualization by semantic substrates. AB - Networks have remained a challenge for information visualization designers because of the complex issues of node and link layout coupled with the rich set of tasks that users present. This paper offers a strategy based on two principles: (1) layouts are based on user-defined semantic substrates, which are non-overlapping regions in which node placement is based on node attributes, (2) users interactively adjust sliders to control link visibility to limit clutter and thus ensure comprehensibility of source and destination. Scalability is further facilitated by user control of which nodes are visible. We illustrate our semantic substrates approach as implemented in NVSS 1.0 with legal precedent data for up to 1122 court cases in three regions with 7645 legal citations. PMID- 17080795 TI - Hierarchical edge bundles: visualization of adjacency relations in hierarchical data. AB - A compound graph is a frequently encountered type of data set. Relations are given between items, and a hierarchy is defined on the items as well. We present a new method for visualizing such compound graphs. Our approach is based on visually bundling the adjacency edges, i.e., non-hierarchical edges, together. We realize this as follows. We assume that the hierarchy is shown via a standard tree visualization method. Next, we bend each adjacency edge, modeled as a B spline curve, toward the polyline defined by the path via the inclusion edges from one node to another. This hierarchical bundling reduces visual clutter and also visualizes implicit adjacency edges between parent nodes that are the result of explicit adjacency edges between their respective child nodes. Furthermore, hierarchical edge bundling is a generic method which can be used in conjunction with existing tree visualization techniques. We illustrate our technique by providing example visualizations and discuss the results based on an informal evaluation provided by potential users of such visualizations. PMID- 17080796 TI - Visualization of geo-spatial point sets via global shape transformation and local pixel placement. AB - In many applications, data is collected and indexed by geo-spatial location. Discovering interesting patterns through visualization is an important way of gaining insight about such data. A previously proposed approach is to apply local placement functions such as PixelMaps that transform the input data set into a solution set that preserves certain constraints while making interesting patterns more obvious and avoid data loss from overplotting. In experience, this family of spatial transformations can reveal fine structures in large point sets, but it is sometimes difficult to relate those structures to basic geographic features such as cities and regional boundaries. Recent information visualization research has addressed other types of transformation functions that make spatially-transformed maps with recognizable shapes. These types of spatial-transformation are called global shape functions. In particular, cartogram-based map distortion has been studied. On the other hand, cartogram-based distortion does not handle point sets readily. In this study, we present a framework that allows the user to specify a global shape function and a local placement function. We combine cartogram-based layout (global shape) with PixelMaps (local placement), obtaining some of the benefits of each toward improved exploration of dense geo-spatial data sets. PMID- 17080797 TI - WORLDMAPPER: the world as you've never seen it before. AB - This paper describes the Worldmapper Project, which makes use of novel visualization techniques to represent a broad variety of social and economic data about the countries of the world. The goal of the project is to use the map projections known as cartograms to depict comparisons and relations between different territories, and its execution raises many interesting design challenges that were not all apparent at the outset. We discuss the approaches taken towards these challenges, some of which may have considerably broad application. We conclude by commenting on the positive initial response to the worldmapper images published on the web, which we believe is due, at least in part, to the particular effectiveness of the cartogram as a tool for communicating quantitative geographic data. PMID- 17080798 TI - Spatial analysis of news sources. AB - People in different places talk about different things. This interest distribution is reflected by the newspaper articles circulated in a particular area. We use data from our large-scale newspaper analysis system (Lydia) to make entity datamaps, a spatial visualization of the interest in a given named entity. Our goal is to identify entities which display regional biases. We develop a model of estimating the frequency of reference of an entity in any given city from the reference frequency centered in surrounding cities, and techniques for evaluating the spatial significance of this distribution. PMID- 17080799 TI - Dynamic map labeling. AB - We address the problem of filtering, selecting and placing labels on a dynamic map, which is characterized by continuous zooming and panning capabilities. This consists of two interrelated issues. The first is to avoid label popping and other artifacts that cause confusion and interrupt navigation, and the second is to label at interactive speed. In most formulations the static map labeling problem is NP-hard, and a fast approximation might have O(nlogn) complexity. Even this is too slow during interaction, when the number of labels shown can be several orders of magnitude less than the number in the map. In this paper we introduce a set of desiderata for "consistent" dynamic map labeling, which has qualities desirable for navigation. We develop a new framework for dynamic labeling that achieves the desiderata and allows for fast interactive display by moving all of the selection and placement decisions into the preprocessing phase. This framework is general enough to accommodate a variety of selection and placement algorithms. It does not appear possible to achieve our desiderata using previous frameworks. Prior to this paper, there were no formal models of dynamic maps or of dynamic labels; our paper introduces both. We formulate a general optimization problem for dynamic map labeling and give a solution to a simple version of the problem. The simple version is based on label priorities and a versatile and intuitive class of dynamic label placements we call "invariant point placements". Despite these restrictions, our approach gives a useful and practical solution. Our implementation is incorporated into the G-Vis system which is a full-detail dynamic map of the continental USA. This demo is available through any browser. PMID- 17080800 TI - Visualization of barrier tree sequences. AB - Dynamical models that explain the formation of spatial structures of RNA molecules have reached a complexity that requires novel visualization methods that help to analyze the validity of these models. Here, we focus on the visualization of so-called folding landscapes of a growing RNA molecule. Folding landscapes describe the energy of a molecule as a function of its spatial configuration; thus they are huge and high dimensional. Their most salient features, however, are encapsulated by their so-called barrier tree that reflects the local minima and their connecting saddle points. For each length of the growing RNA chain there exists a folding landscape. We visualize the sequence of folding landscapes by an animation of the corresponding barrier trees. To generate the animation, we adapt the foresight layout with tolerance algorithm for general dynamic graph layout problems. Since it is very general, we give a detailed description of each phase: constructing a supergraph for the trees, layout of that supergraph using a modified DoT algorithm, and presentation techniques for the final animation. PMID- 17080801 TI - Visualizing business data with generalized treemaps. AB - Business data is often presented using simple business graphics. These familiar visualizations are effective for providing overviews, but fall short for the presentation of large amounts of detailed information. Treemaps can provide such detail, but are often not easy to understand. We present how standard treemap algorithms can be adapted such that the results mimic familiar business graphics. Specifically, we present the use of different layout algorithms per level, a number of variations of the squarified algorithm, the use of variable borders, and the use of non-rectangular shapes. The combined use of these leads to histograms, pie charts and a variety of other styles. PMID- 17080802 TI - FacetMap: A scalable search and browse visualization. AB - The dominant paradigm for searching and browsing large data stores is text-based: presenting a scrollable list of search results in response to textual search term input. While this works well for the Web, there is opportunity for improvement in the domain of personal information stores, which tend to have more heterogeneous data and richer metadata. In this paper, we introduce FacetMap, an interactive, query-driven visualization, generalizable to a wide range of metadata-rich data stores. FacetMap uses a visual metaphor for both input (selection of metadata facets as filters) and output. Results of a user study provide insight into tradeoffs between FacetMap's graphical approach and the traditional text-oriented approach. PMID- 17080803 TI - Visual exploration of complex time-varying graphs. AB - Many graph drawing and visualization algorithms, such as force-directed layout and line-dot rendering, work very well on relatively small and sparse graphs. However, they often produce extremely tangled results and exhibit impractical running times for highly non-planar graphs with large edge density. And very few graph layout algorithms support dynamic time-varying graphs; applying them independently to each frame produces distracting temporally incoherent visualizations. We have developed a new visualization technique based on a novel approach to hierarchically structuring dense graphs via stratification. Using this structure, we formulate a hierarchical force-directed layout algorithm that is both efficient and produces quality graph layouts. The stratification of the graph also allows us to present views of the data that abstract away many small details of its structure. Rather than displaying all edges and nodes at once, resulting in a convoluted rendering, we present an interactive tool that filters edges and nodes using the graph hierarchy and allows users to drill down into the graph for details. Our layout algorithm also accommodates time-varying graphs in a natural way, producing a temporally coherent animation that can be used to analyze and extract trends from dynamic graph data. For example, we demonstrate the use of our method to explore financial correlation data for the U.S. stock market in the period from 1990 to 2005. The user can easily analyze the time varying correlation graph of the market, uncovering information such as market sector trends, representative stocks for portfolio construction, and the interrelationship of stocks over time. PMID- 17080804 TI - Smashing peacocks further: drawing quasi-trees from biconnected components. AB - Quasi-trees, namely graphs with tree-like structure, appear in many application domains, including bioinformatics and computer networks. Our new SPF approach exploits the structure of these graphs with a two-level approach to drawing, where the graph is decomposed into a tree of biconnected components. The low level biconnected components are drawn with a force-directed approach that uses a spanning tree skeleton as a starting point for the layout. The higher-level structure of the graph is a true tree with meta-nodes of variable size that contain each biconnected component. That tree is drawn with a new area-aware variant of a tree drawing algorithm that handles high-degree nodes gracefully, at the cost of allowing edge-node overlaps. SPF performs an order of magnitude faster than the best previous approaches, while producing drawings of commensurate or improved quality. PMID- 17080805 TI - IPSEP-COLA: an incremental procedure for separation constraint layout of graphs. AB - We extend the popular force-directed approach to network (or graph) layout to allow separation constraints, which enforce a minimum horizontal or vertical separation between selected pairs of nodes. This simple class of linear constraints is expressive enough to satisfy a wide variety of application specific layout requirements, including: layout of directed graphs to better show flow; layout with non-overlapping node labels; and layout of graphs with grouped nodes (called clusters). In the stress majorization force-directed layout process, separation constraints can be treated as a quadratic programming problem. We give an incremental algorithm based on gradient projection for efficiently solving this problem. The algorithm is considerably faster than using generic constraint optimization techniques and is comparable in speed to unconstrained stress majorization. We demonstrate the utility of our technique with sample data from a number of practical applications including gene activation networks, terrorist networks and visualization of high-dimensional data. PMID- 17080806 TI - User interaction with scatterplots on small screens--A comparative evaluation of geometric-semantic zoom and fisheye distortion. AB - Existing information-visualization techniques that target small screens are usually limited to exploring a few hundred items. In this article we present a scatterplot tool for Personal Digital Assistants that allows the handling of many thousands of items. The application's scalability is achieved by incorporating two alternative interaction techniques: a geometric-semantic zoom that provides smooth transition between overview and detail, and a fisheye distortion that displays the focus and context regions of the scatterplot in a single view. A user study with 24 participants was conducted to compare the usability and efficiency of both techniques when searching a book database containing 7500 items. The study was run on a pen-driven Wacom board simulating a PDA interface. While the results showed no significant difference in task-completion times, a clear majority of 20 users preferred the fisheye view over the zoom interaction. In addition, other dependent variables such as user satisfaction and subjective rating of orientation and navigation support revealed a preference for the fisheye distortion. These findings partly contradict related research and indicate that, when using a small screen, users place higher value on the ability to preserve navigational context than they do on the ease of use of a simplistic, metaphor-based interaction style. PMID- 17080807 TI - The perceptual scalability of visualization. AB - Larger, higher resolution displays can be used to increase the scalability of information visualizations. But just how much can scalability increase using larger displays before hitting human perceptual or cognitive limits? Are the same visualization techniques that are good on a single monitor also the techniques that are best when they are scaled up using large, high-resolution displays? To answer these questions we performed a controlled experiment on user performance time, accuracy, and subjective workload when scaling up data quantity with different space-time-attribute visualizations using a large, tiled display. Twelve college students used small multiples, embedded bar matrices, and embedded time-series graphs either on a 2 megapixel (Mp) display or with data scaled up using a 32 Mp tiled display. Participants performed various overview and detail tasks on geospatially-referenced multidimensional time-series data. Results showed that current designs are perceptually scalable because they result in a decrease in task completion time when normalized per number of data attributes along with no decrease in accuracy. It appears that, for the visualizations selected for this study, the relative comparison between designs is generally consistent between display sizes. However, results also suggest that encoding is more important on a smaller display while spatial grouping is more important on a larger display. Some suggestions for designers are provided based on our experience designing visualizations for large displays. PMID- 17080808 TI - Complex logarithmic views for small details in large contexts. AB - Commonly known detail in context techniques for the two-dimensional Euclidean space enlarge details and shrink their context using mapping functions that introduce geometrical compression. This makes it difficult or even impossible to recognize shapes for large differences in magnification factors. In this paper we propose to use the complex logarithm and the complex root functions to show very small details even in very large contexts. These mappings are conformal, which means they only locally rotate and scale, thus keeping shapes intact and recognizable. They allow showing details that are orders of magnitude smaller than their surroundings in combination with their context in one seamless visualization. We address the utilization of this universal technique for the interaction with complex two-dimensional data considering the exploration of large graphs and other examples. PMID- 17080809 TI - Software design patterns for information visualization. AB - Despite a diversity of software architectures supporting information visualization, it is often difficult to identify, evaluate, and re-apply the design solutions implemented within such frameworks. One popular and effective approach for addressing such difficulties is to capture successful solutions in design patterns, abstract descriptions of interacting software components that can be customized to solve design problems within a particular context. Based upon a review of existing frameworks and our own experiences building visualization software, we present a series of design patterns for the domain of information visualization. We discuss the structure, context of use, and interrelations of patterns spanning data representation, graphics, and interaction. By representing design knowledge in a reusable form, these patterns can be used to facilitate software design, implementation, and evaluation, and improve developer education and communication. PMID- 17080810 TI - A pipeline for computer aided polyp detection. AB - We present a novel pipeline for computer-aided detection (CAD) of colonic polyps by integrating texture and shape analysis with volume rendering and conformal colon flattening. Using our automatic method, the 3D polyp detection problem is converted into a 2D pattern recognition problem. The colon surface is first segmented and extracted from the CT data set of the patient's abdomen, which is then mapped to a 2D rectangle using conformal mapping. This flattened image is rendered using a direct volume rendering technique with a translucent electronic biopsy transfer function. The polyps are detected by a 2D clustering method on the flattened image. The false positives are further reduced by analyzing the volumetric shape and texture features. Compared with shape based methods, our method is much more efficient without the need of computing curvature and other shape parameters for the whole colon surface. The final detection results are stored in the 2D image, which can be easily incorporated into a virtual colonoscopy (VC) system to highlight the polyp locations. The extracted colon surface mesh can be used to accelerate the volumetric ray casting algorithm used to generate the VC endoscopic view. The proposed automatic CAD pipeline is incorporated into an interactive VC system, with a goal of helping radiologists detect polyps faster and with higher accuracy. PMID- 17080811 TI - Full body virtual autopsies using a state-of-the-art volume rendering pipeline. AB - This paper presents a procedure for virtual autopsies based on interactive 3D visualizations of large scale, high resolution data from CT-scans of human cadavers. The procedure is described using examples from forensic medicine and the added value and future potential of virtual autopsies is shown from a medical and forensic perspective. Based on the technical demands of the procedure state of-the-art volume rendering techniques are applied and refined to enable real time, full body virtual autopsies involving gigabyte sized data on standard GPUs. The techniques applied include transfer function based data reduction using level of-detail selection and multi-resolution rendering techniques. The paper also describes a data management component for large, out-of-core data sets and an extension to the GPU-based raycaster for efficient dual TF rendering. Detailed benchmarks of the pipeline are presented using data sets from forensic cases. PMID- 17080812 TI - Real-time illustration of vascular structures. AB - We present real-time vascular visualization methods, which extend on illustrative rendering techniques to particularly accentuate spatial depth and to improve the perceptive separation of important vascular properties such as branching level and supply area. The resulting visualization can and has already been used for direct projection on a patient's organ in the operation theater where the varying absorption and reflection characteristics of the surface limit the use of color. The important contributions of our work are a GPU-based hatching algorithm for complex tubular structures that emphasizes shape and depth as well as GPU accelerated shadow-like depth indicators, which enable reliable comparisons of depth distances in a static monoscopic 3D visualization. In addition, we verify the expressiveness of our illustration methods in a large, quantitative study with 160 subjects. PMID- 17080813 TI - Lines of curvature for polyp detection in virtual colonoscopy. AB - Computer-aided diagnosis (CAD) is a helpful addition to laborious visual inspection for preselection of suspected colonic polyps in virtual colonoscopy. Most of the previous work on automatic polyp detection makes use of indicators based on the scalar curvature of the colon wall and can result in many false positive detections. Our work tries to reduce the number of false-positive detections in the preselection of polyp candidates. Polyp surface shape can be characterized and visualized using lines of curvature. In this paper, we describe techniques for generating and rendering lines of curvature on surfaces and we show that these lines can be used as part of a polyp detection approach. We have adapted existing approaches on explicit triangular surface meshes, and developed a new algorithm on implicit surfaces embedded in 3D volume data. The visualization of shaded colonic surfaces can be enhanced by rendering the derived lines of curvature on these surfaces. Features strongly correlated with true positive detections were calculated on lines of curvature and used for the polyp candidate selection. We studied the performance of these features on 5 data sets that included 331 pre-detected candidates, of which 50 sites were true polyps. The winding angle had a significant discriminating power for true-positive detections, which was demonstrated by a Wilcoxon rank sum test with p < 0.001. The median winding angle and inter-quartile range (IQR) for true polyps were 7.817 and 6.770 - 9.288 compared to 2.954 and 1.995 - 3.749 for false-positive detections. PMID- 17080814 TI - Outlier-preserving focus+context visualization in parallel coordinates. AB - Focus+context visualization integrates a visually accentuated representation of selected data items in focus (more details, more opacity, etc.) with a visually deemphasized representation of the rest of the data, i.e., the context. The role of context visualization is to provide an overview of the data for improved user orientation and improved navigation. A good overview comprises the representation of both outliers and trends. Up to now, however, context visualization not really treated outliers sufficiently. In this paper we present a new approach to focus+context visualization in parallel coordinates which is truthful to outliers in the sense that small-scale features are detected before visualization and then treated specially during context visualization. Generally, we present a solution which enables context visualization at several levels of abstraction, both for the representation of outliers and trends. We introduce outlier detection and context generation to parallel coordinates on the basis of a binned data representation. This leads to an output-oriented visualization approach which means that only those parts of the visualization process are executed which actually affect the final rendering. Accordingly, the performance of this solution is much more dependent on the visualization size than on the data size which makes it especially interesting for large datasets. Previous approaches are outperformed, the new solution was successfully applied to datasets with up to 3 million data records and up to 50 dimensions. PMID- 17080815 TI - Composite rectilinear deformation for stretch and squish navigation. AB - We present the first scalable algorithm that supports the composition of successive rectilinear deformations. Earlier systems that provided stretch and squish navigation could only handle small datasets. More recent work featuring rubber sheet navigation for large datasets has focused on rendering and on application-specific issues. However, no algorithm has yet been presented for carrying out such navigation methods; our paper addresses this problem. For maximum flexibility with large datasets, a stretch and squish navigation algorithm should allow for millions of potentially deformable regions. However, typical usage only changes the extents of a small subset k of these n regions at a time. The challenge is to avoid computations that are linear in n, because a single deformation can affect the absolute screen-space location of every deformable region. We provide an O(klogn) algorithm that supports any application that can lay out a dataset on a generic grid, and show an implementation that allows navigation of trees and gene sequences with millions of items in sub millisecond time. PMID- 17080816 TI - Multi-variate, time-varying, and comparative visualization with contextual cues. AB - Time-varying, multi-variate, and comparative data sets are not easily visualized due to the amount of data that is presented to the user at once. By combining several volumes together with different operators into one visualized volume, the user is able to compare values from different data sets in space over time, run, or field without having to mentally switch between different renderings of individual data sets. In this paper, we propose using a volume shader where the user is given the ability to easily select and operate on many data volumes to create comparison relationships. The user specifies an expression with set and numerical operations and her data to see relationships between data fields. Furthermore, we render the contextual information of the volume shader by converting it to a volume tree. We visualize the different levels and nodes of the volume tree so that the user can see the results of suboperations. This gives the user a deeper understanding of the final visualization, by seeing how the parts of the whole are operationally constructed. PMID- 17080817 TI - Multifield-graphs: an approach to visualizing correlations in multifield scalar data. AB - We present an approach to visualizing correlations in 3D multifield scalar data. The core of our approach is the computation of correlation fields, which are scalar fields containing the local correlations of subsets of the multiple fields. While the visualization of the correlation fields can be done using standard 3D volume visualization techniques, their huge number makes selection and handling a challenge. We introduce the Multifield-Graph to give an overview of which multiple fields correlate and to show the strength of their correlation. This information guides the selection of informative correlation fields for visualization. We use our approach to visually analyze a number of real and synthetic multifield datasets. PMID- 17080818 TI - Saliency-guided enhancement for volume visualization. AB - Recent research in visual saliency has established a computational measure of perceptual importance. In this paper we present a visual-saliency-based operator to enhance selected regions of a volume. We show how we use such an operator on a user-specified saliency field to compute an emphasis field. We further discuss how the emphasis field can be integrated into the visualization pipeline through its modifications of regional luminance and chrominance. Finally, we validate our work using an eye-tracking-based user study and show that our new saliency enhancement operator is more effective at eliciting viewer attention than the traditional Gaussian enhancement operator. PMID- 17080819 TI - Importance-driven focus of attention. AB - This paper introduces a concept for automatic focusing on features within a volumetric data set. The user selects a focus, i.e., object of interest, from a set of pre-defined features. Our system automatically determines the most expressive view on this feature. A characteristic viewpoint is estimated by a novel information-theoretic framework which is based on the mutual information measure. Viewpoints change smoothly by switching the focus from one feature to another one. This mechanism is controlled by changes in the importance distribution among features in the volume. The highest importance is assigned to the feature in focus. Apart from viewpoint selection, the focusing mechanism also steers visual emphasis by assigning a visually more prominent representation. To allow a clear view on features that are normally occluded by other parts of the volume, the focusing for example incorporates cut-away views. PMID- 17080820 TI - ClearView: An interactive context preserving hotspot visualization technique. AB - Volume rendered imagery often includes a barrage of 3D information like shape, appearance and topology of complex structures, and it thus quickly overwhelms the user. In particular, when focusing on a specific region a user cannot observe the relationship between various structures unless he has a mental picture of the entire data. In this paper we present ClearView, a GPU-based, interactive framework for texture-based volume ray-casting that allows users which do not have the visualization skills for this mental exercise to quickly obtain a picture of the data in a very intuitive and user-friendly way. ClearView is designed to enable the user to focus on particular areas in the data while preserving context information without visual clutter. ClearView does not require additional feature volumes as it derives any features in the data from image information only. A simple point-and-click interface enables the user to interactively highlight structures in the data. ClearView provides an easy to use interface to complex volumetric data as it only uses transparency in combination with a few specific shaders to convey focus and context information. PMID- 17080821 TI - Visualization tools for vorticity transport analysis in incompressible flow. AB - Vortices are undesirable in many applications while indispensable in others. It is therefore of common interest to understand their mechanisms of creation. This paper aims at analyzing the transport of vorticity inside incompressible flow. The analysis is based on the vorticity equation and is performed along pathlines which are typically started in upstream direction from vortex regions. Different methods for the quantitative and explorative analysis of vorticity transport are presented and applied to CFD simulations of water turbines. Simulation quality is accounted for by including the errors of meshing and convergence into analysis and visualization. The obtained results are discussed and interpretations with respect to engineering questions are given. PMID- 17080822 TI - Vortex visualization for practical engineering applications. AB - In order to understand complex vortical flows in large data sets, we must be able to detect and visualize vortices in an automated fashion. In this paper, we present a feature-based vortex detection and visualization technique that is appropriate for large computational fluid dynamics data sets computed on unstructured meshes. In particular, we focus on the application of this technique to visualization of the flow over a serrated wing and the flow field around a spinning missile with dithering canards. We have developed a core line extraction technique based on the observation that vortex cores coincide with local extrema in certain scalar fields. We also have developed a novel technique to handle complex vortex topology that is based on k-means clustering. These techniques facilitate visualization of vortices in simulation data that may not be optimally resolved or sampled. Results are included that highlight the strengths and weaknesses of our approach. We conclude by describing how our approach can be improved to enhance robustness and expand its range of applicability. PMID- 17080823 TI - An advanced evenly-spaced streamline placement algorithm. AB - This paper presents an advanced evenly-spaced streamline placement algorithm for fast, high-quality, and robust layout of flow lines. A fourth-order Runge-Kutta integrator with adaptive step size and error control is employed for rapid accurate streamline advection. Cubic Hermite polynomial interpolation with large sample-spacing is adopted to create fewer evenly-spaced samples along each streamline to reduce the amount of distance checking. We propose two methods to enhance placement quality. Double queues are used to prioritize topological seeding and to favor long streamlines to minimize discontinuities. Adaptive distance control based on the local flow variance is explored to reduce cavities. Furthermore, we propose a universal, effective, fast, and robust loop detection strategy to address closed and spiraling streamlines. Our algorithm is an order of-magnitude faster than Jobard and Lefer's algorithm with better placement quality and over 5 times faster than Mebarki et al.'s algorithm with comparable placement quality, but with a more robust solution to loop detection. PMID- 17080824 TI - Fine-grained visualization pipelines and lazy functional languages. AB - The pipeline model in visualization has evolved from a conceptual model of data processing into a widely used architecture for implementing visualization systems. In the process, a number of capabilities have been introduced, including streaming of data in chunks, distributed pipelines, and demand-driven processing. Visualization systems have invariably built on stateful programming technologies, and these capabilities have had to be implemented explicitly within the lower layers of a complex hierarchy of services. The good news for developers is that applications built on top of this hierarchy can access these capabilities without concern for how they are implemented. The bad news is that by freezing capabilities into low-level services expressive power and flexibility is lost. In this paper we express visualization systems in a programming language that more naturally supports this kind of processing model. Lazy functional languages support fine-grained demand-driven processing, a natural form of streaming, and pipeline-like function composition for assembling applications. The technology thus appears well suited to visualization applications. Using surface extraction algorithms as illustrative examples, and the lazy functional language Haskell, we argue the benefits of clear and concise expression combined with fine-grained, demand-driven computation. Just as visualization provides insight into data, functional abstraction provides new insight into visualization. PMID- 17080825 TI - A novel visualization model for web search results. AB - This paper presents an interactive visualization system, named WebSearchViz, for visualizing the Web search results and acilitating users' navigation and exploration. The metaphor in our model is the solar system with its planets and asteroids revolving around the sun. Location, color, movement, and spatial distance of objects in the visual space are used to represent the semantic relationships between a query and relevant Web pages. Especially, the movement of objects and their speeds add a new dimension to the visual space, illustrating the degree of relevance among a query and Web search results in the context of users' subjects of interest. By interacting with the visual space, users are able to observe the semantic relevance between a query and a resulting Web page with respect to their subjects of interest, context information, or concern. Users' subjects of interest can be dynamically changed, redefined, added, or deleted from the visual space. PMID- 17080826 TI - A trajectory-preserving synchronization method for collaborative visualization. AB - In the past decade, a lot of research work has been conducted to support collaborative visualization among remote users over the networks, allowing them to visualize and manipulate shared data for problem solving. There are many applications of collaborative visualization, such as oceanography, meteorology and medical science. To facilitate user interaction, a critical system requirement for collaborative visualization is to ensure that remote users will perceive a synchronized view of the shared data. Failing this requirement, the user's ability in performing the desirable collaborative tasks will be affected. In this paper, we propose a synchronization method to support collaborative visualization. It considers how interaction with dynamic objects is perceived by application participants under the existence of network latency, and remedies the motion trajectory of the dynamic objects. It also handles the false positive and false negative collision detection problems. The new method is particularly well designed for handling content changes due to unpredictable user interventions or object collisions. We demonstrate the effectiveness of our method through a number of experiments. PMID- 17080827 TI - Concurrent visualization in a production supercomputing environment. AB - We describe a concurrent visualization pipeline designed for operation in a production supercomputing environment. The facility was initially developed on the NASA Ames "Columbia" supercomputer for a massively parallel forecast model (GEOS4). During the 2005 Atlantic hurricane season, GEOS4 was run 4 times a day under tight time constraints so that its output could be included in an ensemble prediction that was made available to forecasters at the National Hurricane Center. Given this time-critical context, we designed a configurable concurrent pipeline to visualize multiple global fields without significantly affecting the runtime model performance or reliability. We use MPEG compression of the accruing images to facilitate live low-bandwidth distribution of multiple visualization streams to remote sites. We also describe the use of our concurrent visualization framework with a global ocean circulation model, which provides a 864-fold increase in the temporal resolution of practically achievable animations. In both the atmospheric and oceanic circulation models, the application scientists gained new insights into their model dynamics, due to the high temporal resolution animations attainable. PMID- 17080828 TI - Scalable WIM: effective exploration in large-scale astrophysical environments. AB - Navigating through large-scale virtual environments such as simulations of the astrophysical Universe is difficult. The huge spatial range of astronomical models and the dominance of empty space make it hard for users to travel across cosmological scales effectively, and the problem of wayfinding further impedes the user's ability to acquire reliable spatial knowledge of astronomical contexts. We introduce a new technique called the scalable world-in-miniature (WIM) map as a unifying interface to facilitate travel and wayfinding in a virtual environment spanning gigantic spatial scales: Power-law spatial scaling enables rapid and accurate transitions among widely separated regions; logarithmically mapped miniature spaces offer a global overview mode when the full context is too large; 3D landmarks represented in the WIM are enhanced by scale, positional, and directional cues to augment spatial context awareness; a series of navigation models are incorporated into the scalable WIM to improve the performance of travel tasks posed by the unique characteristics of virtual cosmic exploration. The scalable WIM user interface supports an improved physical navigation experience and assists pragmatic cognitive understanding of a visualization context that incorporates the features of large-scale astronomy. PMID- 17080829 TI - Using visual cues of contact to improve interactive manipulation of virtual objects in industrial assembly/maintenance simulations. AB - This paper describes a set of visual cues of contact designed to improve the interactive manipulation of virtual objects in industrial assembly/maintenance simulations. These visual cues display information of proximity, contact and effort between virtual objects when the user manipulates a part inside a digital mock-up. The set of visual cues encloses the apparition of glyphs (arrow, disk, or sphere) when the manipulated object is close or in contact with another part of the virtual environment. Light sources can also be added at the level of contact points. A filtering technique is proposed to decrease the number of glyphs displayed at the same time. Various effects--such as change in color, change in size, and deformation of shape- can be applied to the glyphs as a function of proximity with other objects or amplitude of the contact forces. A preliminary evaluation was conducted to gather the subjective preference of a group of participants during the simulation of an automotive assembly operation. The collected questionnaires showed that participants globally appreciated our visual cues of contact. The changes in color appeared to be preferred concerning the display of distances and proximity information. Size changes and deformation effects appeared to be preferred in terms of perception of contact forces between the parts. Last, light sources were selected to focus the attention of the user on the contact areas. PMID- 17080830 TI - High-level user interfaces for transfer function design with semantics. AB - Many sophisticated techniques for the visualization of volumetric data such as medical data have been published. While existing techniques are mature from a technical point of view, managing the complexity of visual parameters is still difficult for non-expert users. To this end, this paper presents new ideas to facilitate the specification of optical properties for direct volume rendering. We introduce an additional level of abstraction for parametric models of transfer functions. The proposed framework allows visualization experts to design high level transfer function models which can intuitively be used by non-expert users. The results are user interfaces which provide semantic information for specialized visualization problems. The proposed method is based on principal component analysis as well as on concepts borrowed from computer animation. PMID- 17080831 TI - LOD map--A visual interface for navigating multiresolution volume visualization. AB - In multiresolution volume visualization, a visual representation of level-of detail (LOD) quality is important for us to examine, compare, and validate different LOD selection algorithms. While traditional methods rely on ultimate images for quality measurement, we introduce the LOD map--an alternative representation of LOD quality and a visual interface for navigating multiresolution data exploration. Our measure for LOD quality is based on the formulation of entropy from information theory. The measure takes into account the distortion and contribution of multiresolution data blocks. A LOD map is generated through the mapping of key LOD ingredients to a treemap representation. The ordered treemap layout is used for relative stable update of the LOD map when the view or LOD changes. This visual interface not only indicates the quality of LODs in an intuitive way, but also provides immediate suggestions for possible LOD improvement through visually-striking features. It also allows us to compare different views and perform rendering budget control. A set of interactive techniques is proposed to make the LOD adjustment a simple and easy task. We demonstrate the effectiveness and efficiency of our approach on large scientific and medical data sets. PMID- 17080832 TI - Analyzing complex FTMS simulations: a case study in high-level visualization of ion motions. AB - Current practice in particle visualization renders particle position data directly onto the screen as points or glyphs. Using a camera placed at a fixed position, particle motions can be visualized by rendering trajectories or by animations. Applying such direct techniques to large, time dependent particle data sets often results in cluttered images in which the dynamic properties of the underlying system are difficult to interpret. In this case study we take an alternative approach to the visualization of ion motions. Instead of rendering ion position data directly, we first extract meaningful motion information from the ion position data and then map this information onto geometric primitives. Our goal is to produce high-level visualizations that reflect the physicists' way of thinking about ion dynamics. Parameterized geometric icons are defined to encode motion information of clusters of related ions. In addition, a parameterized camera control mechanism is used to analyze relative instead of only absolute ion motions. We apply the techniques to simulations of Fourier transform mass spectrometry (FTMS) experiments. The data produced by such simulations can amount to 5 10(4) ions and 10(5) timesteps. This paper discusses the requirements, design and informal evaluation of the implemented system. PMID- 17080833 TI - Detection and visualization of defects in 3D unstructured models of nematic liquid crystals. AB - A method for the semi-automatic detection and visualization of defects in models of nematic liquid crystals (NLCs) is introduced; this method is suitable for unstructured models, a previously unsolved problem. The detected defects-also known as disclinations-are regions were the alignment of the liquid crystal rapidly changes over space; these defects play a large role in the physical behavior of the NLC substrate. Defect detection is based upon a measure of total angular change of crystal orientation (the director) over a node neighborhood via the use of a nearest neighbor path. Visualizations based upon the detection algorithm clearly identifies complete defect regions as opposed to incomplete visual descriptions provided by cutting-plane and isosurface approaches. The introduced techniques are currently in use by scientists studying the dynamics of defect change. PMID- 17080834 TI - Understanding the structure of the turbulent mixing layer in hydrodynamic instabilities. AB - When a heavy fluid is placed above a light fluid, tiny vertical perturbations in the interface create a characteristic structure of rising bubbles and falling spikes known as Rayleigh-Taylor instability. Rayleigh-Taylor instabilities have received much attention over the past half-century because of their importance in understanding many natural and man-made phenomena, ranging from the rate of formation of heavy elements in supernovae to the design of capsules for Inertial Confinement Fusion. We present a new approach to analyze Rayleigh-Taylor instabilities in which we extract a hierarchical segmentation of the mixing envelope surface to identify bubbles and analyze analogous segmentations of fields on the original interface plane. We compute meaningful statistical information that reveals the evolution of topological features and corroborates the observations made by scientists. We also use geometric tracking to follow the evolution of single bubbles and highlight merge/split events leading to the formation of the large and complex structures characteristic of the later stages. In particular we (i) Provide a formal definition of a bubble; (ii) Segment the envelope surface to identify bubbles; (iii) Provide a multi-scale analysis technique to produce statistical measures of bubble growth; (iv) Correlate bubble measurements with analysis of fields on the interface plane; (v) Track the evolution of individual bubbles over time. Our approach is based on the rigorous mathematical foundations of Morse theory and can be applied to a more general class of applications. PMID- 17080835 TI - Hub-based simulation and graphics hardware accelerated visualization for nanotechnology applications. AB - The Network for Computational Nanotechnology (NCN) has developed a science gateway at nanoHUB.org for nanotechnology education and research. Remote users can browse through online seminars and courses, and launch sophisticated nanotechnology simulation tools, all within their web browser. Simulations are supported by a middleware that can route complex jobs to grid supercomputing resources. But what is truly unique about the middleware is the way that it uses hardware accelerated graphics to support both problem setup and result visualization. This paper describes the design and integration of a remote visualization framework into the nanoHUB for interactive visual analytics of nanotechnology simulations. Our services flexibly handle a variety of nanoscience simulations, render them utilizing graphics hardware acceleration in a scalable manner, and deliver them seamlessly through the middleware to the user. Rendering is done only on-demand, as needed, so each graphics hardware unit can simultaneously support many user sessions. Additionally, a novel node distribution scheme further improves our system's scalability. Our approach is not only efficient but also cost-effective. Only a half-dozen render nodes are anticipated to support hundreds of active tool sessions on the nanoHUB. Moreover, this architecture and visual analytics environment provides capabilities that can serve many areas of scientific simulation and analysis beyond nanotechnology with its ability to interactively analyze and visualize multivariate scalar and vector fields. PMID- 17080836 TI - Feature aligned volume manipulation for illustration and visualization. AB - In this paper we describe a GPU-based technique for creating illustrative visualization through interactive manipulation of volumetric models. It is partly inspired by medical illustrations, where it is common to depict cuts and deformation in order to provide a better understanding of anatomical and biological structures or surgical processes, and partly motivated by the need for a real-time solution that supports the specification and visualization of such illustrative manipulation. We propose two new feature-aligned techniques, namely surface alignment and segment alignment, and compare them with the axis-aligned techniques which was reported in previous work on volume manipulation. We also present a mechanism for defining features using texture volumes, and methods for computing correct normals for the deformed volume in respect to different alignments. We describe a GPU-based implementation to achieve real-time performance of the techniques and a collection of manipulation operators including peelers, retractors, pliers and dilators which are adaptations of the metaphors and tools used in surgical procedures and medical illustrations. Our approach is directly applicable in medical and biological illustration, and we demonstrate how it works as an interactive tool for focus+context visualization, as well as a generic technique for volume graphics. PMID- 17080837 TI - Exploded views for volume data. AB - Exploded views are an illustration technique where an object is partitioned into several segments. These segments are displaced to reveal otherwise hidden detail. In this paper we apply the concept of exploded views to volumetric data in order to solve the general problem of occlusion. In many cases an object of interest is occluded by other structures. While transparency or cutaways can be used to reveal a focus object, these techniques remove parts of the context information. Exploded views, on the other hand, do not suffer from this drawback. Our approach employs a force-based model: the volume is divided into a part configuration controlled by a number of forces and constraints. The focus object exerts an explosion force causing the parts to arrange according to the given constraints. We show that this novel and flexible approach allows for a wide variety of explosion-based visualizations including view-dependent explosions. Furthermore, we present a high-quality GPU-based volume ray casting algorithm for exploded views which allows rendering and interaction at several frames per second. PMID- 17080838 TI - Caricaturistic visualization. AB - Caricatures are pieces of art depicting persons or sociological conditions in a non-veridical way. In both cases caricatures are referring to a reference model. The deviations from the reference model are the characteristic features of the depicted subject. Good caricatures exaggerate the characteristics of a subject in order to accent them. The concept of caricaturistic visualization is based on the caricature metaphor. The aim of caricaturistic visualization is an illustrative depiction of characteristics of a given dataset by exaggerating deviations from the reference model. We present the general concept of caricaturistic visualization as well as a variety of examples. We investigate different visual representations for the depiction of caricatures. Further, we present the caricature matrix, a technique to make differences between datasets easily identifiable. PMID- 17080839 TI - Visual signatures in video visualization. AB - Video visualization is a computation process that extracts meaningful information from original video data sets and conveys the extracted information to users in appropriate visual representations. This paper presents a broad treatment of the subject, following a typical research pipeline involving concept formulation, system development, a path-finding user study, and a field trial with real application data. In particular, we have conducted a fundamental study on the visualization of motion events in videos. We have, for the first time, deployed flow visualization techniques in video visualization. We have compared the effectiveness of different abstract visual representations of videos. We have conducted a user study to examine whether users are able to learn to recognize visual signatures of motions, and to assist in the evaluation of different visualization techniques. We have applied our understanding and the developed techniques to a set of application video clips. Our study has demonstrated that video visualization is both technically feasible and cost-effective. It has provided the first set of evidence confirming that ordinary users can be accustomed to the visual features depicted in video visualizations, and can learn to recognize visual signatures of a variety of motion events. PMID- 17080840 TI - Asynchronous distributed calibration for scalable and reconfigurable multi projector displays. AB - Centralized techniques have been used until now when automatically calibrating (both geometrically and photometrically) large high-resolution displays created by tiling multiple projectors in a 2D array. A centralized server managed all the projectors and also the camera(s) used to calibrate the display. In this paper, we propose an asynchronous distributed calibration methodology via a display unit called the plug-and-play projector (PPP). The PPP consists of a projector, camera, computation and communication unit, thus creating a self-sufficient module that enables an asynchronous distributed architecture for multi-projector displays. We present a single-program-multiple-data (SPMD) calibration algorithm that runs on each PPP and achieves a truly scalable and reconfigurable display without any input from the user. It instruments novel capabilities like adding/removing PPPs from the display dynamically, detecting faults, and reshaping the display to a reasonable rectangular shape to react to the addition/removal/faults. To the best of our knowledge, this is the first attempt to realize a completely asynchronous and distributed calibration architecture and methodology for multi-projector displays. PMID- 17080841 TI - Dynamic view selection for time-varying volumes. AB - Animation is an effective way to show how time-varying phenomena evolve over time. A key issue of generating a good animation is to select ideal views through which the user can perceive the maximum amount of information from the time varying dataset. In this paper, we first propose an improved view selection method for static data. The method measures the quality of a static view by analyzing the opacity, color and curvature distributions of the corresponding volume rendering images from the given view. Our view selection metric prefers an even opacity distribution with a larger projection area, a larger area of salient features' colors with an even distribution among the salient features, and more perceived curvatures. We use this static view selection method and a dynamic programming approach to select time-varying views. The time-varying view selection maximizes the information perceived from the time-varying dataset based on the constraints that the time-varying view should show smooth changes of direction and near-constant speed. We also introduce a method that allows the user to generate a smooth transition between any two views in a given time step, with the perceived information maximized as well. By combining the static and dynamic view selection methods, the users are able to generate a time-varying view that shows the maximum amount of information from a time-varying data set. PMID- 17080842 TI - Enhancing depth perception in translucent volumes. AB - We present empirical studies that consider the effects of stereopsis and simulated aerial perspective on depth perception in translucent volumes. We consider a purely absorptive lighting model, in which light is not scattered or reflected, but is simply absorbed as it passes through the volume. A purely absorptive lighting model is used, for example, when rendering digitally reconstructed radiographs (DRRs), which are synthetic X-ray images reconstructed from CT volumes. Surgeons make use of DRRs in planning and performing operations, so an improvement of depth perception in DRRs may help diagnosis and surgical planning. PMID- 17080843 TI - Texturing of layered surfaces for optimal viewing. AB - This paper is a contribution to the literature on perceptually optimal visualizations of layered three-dimensional surfaces. Specifically, we develop guidelines for generating texture patterns, which, when tiled on two overlapped surfaces, minimize confusion in depth-discrimination and maximize the ability to localize distinct features. We design a parameterized texture space and explore this texture space using a "human in the loop" experimental approach. Subjects are asked to rate their ability to identify Gaussian bumps on both upper and lower surfaces of noisy terrain fields. Their ratings direct a genetic algorithm, which selectively searches the texture parameter space to find fruitful areas. Data collected from these experiments are analyzed to determine what combinations of parameters work well and to develop texture generation guidelines. Data analysis methods include ANOVA, linear discriminant analysis, decision trees, and parallel coordinates. To confirm the guidelines, we conduct a post-analysis experiment, where subjects rate textures following our guidelines against textures violating the guidelines. Across all subjects, textures following the guidelines consistently produce high rated textures on an absolute scale, and are rated higher than those that did not follow the guidelines. PMID- 17080844 TI - Subjective quantification of perceptual interactions among some 2D scientific visualization methods. AB - We present an evaluation of a parameterized set of 2D icon-based visualization methods where we quantified how perceptual interactions among visual elements affect effective data exploration. During the experiment, subjects quantified three different design factors for each method: the spatial resolution it could represent, the number of data values it could display at each point, and the degree to which it is visually linear. The class of visualization methods includes Poisson-disk distributed icons where icon size, icon spacing, and icon brightness can be set to a constant or coupled to data values from a 2D scalar field. By only coupling one of those visual components to data, we measured filtering interference for all three design factors. Filtering interference characterizes how different levels of the constant visual elements affect the evaluation of the data-coupled element. Our novel experimental methodology allowed us to generalize this perceptual information, gathered using ad-hoc artificial datasets, onto quantitative rules for visualizing real scientific datasets. This work also provides a framework for evaluating visualizations of multi-valued data that incorporate additional visual cues, such as icon orientation or color. PMID- 17080845 TI - Occlusion-free animation of driving routes for car navigation systems. AB - This paper presents a method for occlusion-free animation of geographical landmarks, and its application to a new type of car navigation system in which driving routes of interest are always visible. This is achieved by animating a nonperspective image where geographical landmarks such as mountain tops and roads are rendered as if they are seen from different viewpoints. The technical contribution of this paper lies in formulating the nonperspective terrain navigation as an inverse problem of continuously deforming a 3D terrain surface from the 2D screen arrangement of its associated geographical landmarks. The present approach provides a perceptually reasonable compromise between the navigation clarity and visual realism where the corresponding nonperspective view is fully augmented by assigning appropriate textures and shading effects to the terrain surface according to its geometry. An eye tracking experiment is conducted to prove that the present approach actually exhibits visually-pleasing navigation frames while users can clearly recognize the shape of the driving route without occlusion, together with the spatial configuration of geographical landmarks in its neighborhood. PMID- 17080846 TI - Interactive visualization of intercluster galaxy structures in the horologium reticulum supercluster. AB - We present GyVe, an interactive visualization tool for understanding structure in sparse three-dimensional (3D) point data. The scientific goal driving the tool's development is to determine the presence of filaments and voids as defined by inferred 3D galaxy positions within the Horologium-Reticulum supercluster (HRS). GyVe provides visualization techniques tailored to examine structures defined by the intercluster galaxies. Specific techniques include: interactive user control to move between a global overview and local viewpoints, labelled axes and curved drop lines to indicate positions in the astronomical RA-DEC-cz coordinate system, torsional rocking and stereo to enhance 3D perception, and geometrically distinct glyphs to show potential correlation between intercluster galaxies and known clusters. We discuss the rationale for each design decision and review the success of the techniques in accomplishing the scientific goals. In practice, GyVe has been useful for gaining intuition about structures that were difficult to perceive with 2D projection techniques alone. For example, during their initial session with GyVe, our collaborators quickly confirmed scientific conclusions regarding the large-scale structure of the HRS previously obtained over months of study with 2D projections and statistical techniques. Further use of GyVe revealed the spherical shape of voids and showed that a presumed filament was actually two disconnected structures. PMID- 17080847 TI - An atmospheric visual analysis and exploration system. AB - Meteorological research involves the analysis of multi-field, multi-scale, and multi-source data sets. In order to better understand these data sets, models and measurements at different resolutions must be analyzed. Unfortunately, traditional atmospheric visualization systems only provide tools to view a limited number of variables and small segments of the data. These tools are often restricted to two-dimensional contour or vector plots or three-dimensional isosurfaces. The meteorologist must mentally synthesize the data from multiple plots to glean the information needed to produce a coherent picture of the weather phenomenon of interest. In order to provide better tools to meteorologists and reduce system limitations, we have designed an integrated atmospheric visual analysis and exploration system for interactive analysis of weather data sets. Our system allows for the integrated visualization of 1D, 2D, and 3D atmospheric data sets in common meteorological grid structures and utilizes a variety of rendering techniques. These tools provide meteorologists with new abilities to analyze their data and answer questions on regions of interest, ranging from physics-based atmospheric rendering to illustrative rendering containing particles and glyphs. In this paper, we will discuss the use and performance of our visual analysis for two important meteorological applications. The first application is warm rain formation in small cumulus clouds. Here, our three-dimensional, interactive visualization of modeled drop trajectories within spatially correlated fields from a cloud simulation has provided researchers with new insight. Our second application is improving and validating severe storm models, specifically the Weather Research and Forecasting (WRF) model. This is done through correlative visualization of WRF model and experimental Doppler storm data. PMID- 17080848 TI - Visualization of fibrous and thread-like data. AB - Thread-like structures are becoming more common in modern volumetric data sets as our ability to image vascular and neural tissue at higher resolutions improves. The thread-like structures of neurons and micro-vessels pose a unique problem in visualization since they tend to be densely packed in small volumes of tissue. This makes it difficult for an observer to interpret useful patterns from the data or trace individual fibers. In this paper we describe several methods for dealing with large amounts of thread-like data, such as data sets collected using Knife-Edge Scanning Microscopy (KESM) and Serial Block-Face Scanning Electron Microscopy (SBF-SEM). These methods allow us to collect volumetric data from embedded samples of whole-brain tissue. The neuronal and microvascular data that we acquire consists of thin, branching structures extending over very large regions. Traditional visualization schemes are not sufficient to make sense of the large, dense, complex structures encountered. In this paper, we address three methods to allow a user to explore a fiber network effectively. We describe interactive techniques for rendering large sets of neurons using self-orienting surfaces implemented on the GPU. We also present techniques for rendering fiber networks in a way that provides useful information about flow and orientation. Third, a global illumination framework is used to create high-quality visualizations that emphasize the underlying fiber structure. Implementation details, performance, and advantages and disadvantages of each approach are discussed. PMID- 17080849 TI - Comparative visualization for wave-based and geometric acoustics. AB - We present a comparative visualization of the acoustic simulation results obtained by two different approaches that were combined into a single simulation algorithm. The first method solves the wave equation on a volume grid based on finite elements. The second method, phonon tracing, is a geometric approach that we have previously developed for interactive simulation, visualization and modeling of room acoustics. Geometric approaches of this kind are more efficient than FEM in the high and medium frequency range. For low frequencies they fail to represent diffraction, which on the other hand can be simulated properly by means of FEM. When combining both methods we need to calibrate them properly and estimate in which frequency range they provide comparable results. For this purpose we use an acoustic metric called gain and display the resulting error. Furthermore we visualize interference patterns, since these depend not only on diffraction, but also exhibit phase-dependent amplification and neutralization effects. PMID- 17080850 TI - Hybrid visualization for white matter tracts using triangle strips and point sprites. AB - Diffusion tensor imaging is of high value in neurosurgery, providing information about the location of white matter tracts in the human brain. For their reconstruction, streamline techniques commonly referred to as fiber tracking model the underlying fiber structures and have therefore gained interest. To meet the requirements of surgical planning and to overcome the visual limitations of line representations, a new real-time visualization approach of high visual quality is introduced. For this purpose, textured triangle strips and point sprites are combined in a hybrid strategy employing GPU programming. The triangle strips follow the fiber streamlines and are textured to obtain a tube-like appearance. A vertex program is used to orient the triangle strips towards the camera. In order to avoid triangle flipping in case of fiber segments where the viewing and segment direction are parallel, a correct visual representation is achieved in these areas by chains of point sprites. As a result, a high quality visualization similar to tubes is provided allowing for interactive multimodal inspection. Overall, the presented approach is faster than existing techniques of similar visualization quality and at the same time allows for real-time rendering of dense bundles encompassing a high number of fibers, which is of high importance for diagnosis and surgical planning. PMID- 17080851 TI - Analyzing vortex breakdown flow structures by assignment of colors to tensor invariants. AB - Topological methods are often used to describe flow structures in fluid dynamics and topological flow field analysis usually relies on the invariants of the associated tensor fields. A visual impression of the local properties of tensor fields is often complex and the search of a suitable technique for achieving this is an ongoing topic in visualization. This paper introduces and assesses a method of representing the topological properties of tensor fields and their respective flow patterns with the use of colors. First, a tensor norm is introduced, which preserves the properties of the tensor and assigns the tensor invariants to values of the RGB color space. Secondly, the RGB colors of the tensor invariants are transferred to corresponding hue values as an alternative color representation. The vectorial tensor invariants field is reduced to a scalar hue field and visualization of iso-surfaces of this hue value field allows us to identify locations with equivalent flow topology. Additionally highlighting by the maximum of the eigenvalue difference field reflects the magnitude of the structural change of the flow. The method is applied on a vortex breakdown flow structure inside a cylinder with a rotating lid. PMID- 17080852 TI - Superellipsoid-based, real symmetric traceless tensor glyphs motivated by nematic liquid crystal alignment visualization. AB - A glyph-based method for visualizing the nematic liquid crystal alignment tensor is introduced. Unlike previous approaches, the glyph is based upon physically linked metrics, not offsets of the eigenvalues. These metrics, combined with a set of superellipsoid shapes, communicate both the strength of the crystal's uniaxial alignment and the amount of biaxiality. With small modifications, our approach can visualize any real symmetric traceless tensor. PMID- 17080853 TI - High-quality extraction of isosurfaces from regular and irregular grids. AB - Isosurfaces are ubiquitous in many fields, including visualization, graphics, and vision. They are often the main computational component of important processing pipelines (e.g. , surface reconstruction), and are heavily used in practice. The classical approach to compute isosurfaces is to apply the Marching Cubes algorithm, which although robust and simple to implement, generates surfaces that require additional processing steps to improve triangle quality and mesh size. An important issue is that in some cases, the surfaces generated by Marching Cubes are irreparably damaged, and important details are lost which can not be recovered by subsequent processing. The main motivation of this work is to develop a technique capable of constructing high-quality and high-fidelity isosurfaces. We propose a new advancing front technique that is capable of creating high-quality isosurfaces from regular and irregular volumetric datasets. Our work extends the guidance field framework of Schreiner et al. to implicit surfaces, and improves it in significant ways. In particular, we describe a set of sampling conditions that guarantee that surface features will be captured by the algorithm. We also describe an efficient technique to compute a minimal guidance field, which greatly improves performance. Our experimental results show that our technique can generate high-quality meshes from complex datasets. PMID- 17080854 TI - Mesh layouts for block-based caches. AB - Current computer architectures employ caching to improve the performance of a wide variety of applications. One of the main characteristics of such cache schemes is the use of block fetching whenever an uncached data element is accessed. To maximize the benefit of the block fetching mechanism, we present novel cache-aware and cache-oblivious layouts of surface and volume meshes that improve the performance of interactive visualization and geometric processing algorithms. Based on a general I/O model, we derive new cache-aware and cache oblivious metrics that have high correlations with the number of cache misses when accessing a mesh. In addition to guiding the layout process, our metrics can be used to quantify the quality of a layout, e.g. for comparing different layouts of the same mesh and for determining whether a given layout is amenable to significant improvement. We show that layouts of unstructured meshes optimized for our metrics result in improvements over conventional layouts in the performance of visualization applications such as isosurface extraction and view dependent rendering. Moreover, we improve upon recent cache-oblivious mesh layouts in terms of performance, applicability, and accuracy. PMID- 17080855 TI - Out-of-Core remeshing of large polygonal meshes. AB - We propose an out-of-core method for creating semi-regular surface representations from large input surface meshes. Our approach is based on a streaming implementation of the MAPS remesher of Lee et al. [18]. Our remeshing procedure consists of two stages. First, a simplification process is used to obtain the base domain. During simplification, we maintain the mapping information between the input and the simplified meshes. The second stage of remeshing uses the mapping information to produce samples of the output semi regular mesh. The out-of-core operation of our method is enabled by the synchronous streaming of a simplified mesh and the mapping information stored at the original vertices. The synchronicity of two streaming buffers is maintained using a specially designed write strategy for each buffer. Experimental results demonstrate the remeshing performance of the proposed method, as well as other applications that use the created mapping between the simplified and the original surface representations. PMID- 17080856 TI - Interactive point-based rendering of higher-order tetrahedral data. AB - Computational simulations frequently generate solutions defined over very large tetrahedral volume meshes containing many millions of elements. Furthermore, such solutions may often be expressed using non-linear basis functions. Certain solution techniques, such as discontinuous Galerkin methods, may even produce non conforming meshes. Such data is difficult to visualize interactively, as it is far too large to fit in memory and many common data reduction techniques, such as mesh simplification, cannot be applied to non-conforming meshes. We introduce a point-based visualization system for interactive rendering of large, potentially non-conforming, tetrahedral meshes. We propose methods for adaptively sampling points from non-linear solution data and for decimating points at run time to fit GPU memory limits. Because these are streaming processes, memory consumption is independent of the input size. We also present an order-independent point rendering method that can efficiently render volumes on the order of 20 million tetrahedra at interactive rates. PMID- 17080857 TI - Ambient occlusion and edge cueing to enhance real time molecular visualization. AB - The paper presents a set of combined techniques to enhance the real-time visualization of simple or complex molecules (up to order of 106 atoms) space fill mode. The proposed approach includes an innovative technique for efficient computation and storage of ambient occlusion terms, a small set of GPU accelerated procedural impostors for space-fill and ball-and-stick rendering, and novel edge-cueing techniques. As a result, the user's understanding of the three dimensional structure under inspection is strongly increased (even for still images), while the rendering still occurs in real time. PMID- 17080858 TI - Fast and efficient compression of floating-point data. AB - Large scale scientific simulation codes typically run on a cluster of CPUs that write/read time steps to/from a single file system. As data sets are constantly growing in size, this increasingly leads to I/O bottlenecks. When the rate at which data is produced exceeds the available I/O bandwidth, the simulation stalls and the CPUs are idle. Data compression can alleviate this problem by using some CPU cycles to reduce the amount of data needed to be transfered. Most compression schemes, however, are designed to operate offline and seek to maximize compression, not throughput. Furthermore, they often require quantizing floating point values onto a uniform integer grid, which disqualifies their use in applications where exact values must be retained. We propose a simple scheme for lossless, online compression of floating-point data that transparently integrates into the I/O of many applications. A plug-in scheme for data-dependent prediction makes our scheme applicable to a wide variety of data used in visualization, such as unstructured meshes, point sets, images, and voxel grids. We achieve state-of the-art compression rates and speeds, the latter in part due to an improved entropy coder. We demonstrate that this significantly accelerates I/O throughput in real simulation runs. Unlike previous schemes, our method also adapts well to variable-precision floating-point and integer data. PMID- 17080859 TI - Isualization and analysis of large data collections: a case study applied to confocal microscopy data. AB - In this paper we propose an approach in which interactive visualization and analysis are combined with batch tools for the processing of large data collections. Large and heterogeneous data collections are difficult to analyze and pose specific problems to interactive visualization. Application of the traditional interactive processing and visualization approaches as well as batch processing encounter considerable drawbacks for such large and heterogeneous data collections due to the amount and type of data. Computing resources are not sufficient for interactive exploration of the data and automated analysis has the disadvantage that the user has only limited control and feedback on the analysis process. In our approach, an analysis procedure with features and attributes of interest for the analysis is defined interactively. This procedure is used for off-line processing of large collections of data sets. The results of the batch process along with "visual summaries" are used for further analysis. Visualization is not only used for the presentation of the result, but also as a tool to monitor the validity and quality of the operations performed during the batch process. Operations such as feature extraction and attribute calculation of the collected data sets are validated by visual inspection. This approach is illustrated by an extensive case study, in which a collection of confocal microscopy data sets is analyzed. PMID- 17080860 TI - On histograms and isosurface statistics. AB - In this paper, we show that histograms represent spatial function distributions with a nearest neighbour interpolation. We confirm that this results in systematic underrepresentation of transitional features of the data, and provide new insight why this occurs. We further show that isosurface statistics, which use higher quality interpolation, give better representations of the function distribution. We also use our experimentally collected isosurface statistics to resolve some questions as to the formal complexity of isosurfaces. PMID- 17080861 TI - Interactive point-based isosurface exploration and high-quality rendering. AB - We present an efficient point-based isosurface exploration system with high quality rendering. Our system incorporates two point-based isosurface extraction and visualization methods: edge splatting and the edge kernel method. In a volume, two neighboring voxels define an edge. The intersection points between the active edges and the isosurface are used for exact isosurface representation. The point generation is incorporated in the GPU-based hardware-accelerated rendering, thus avoiding any overhead when changing the isovalue in the exploration. We call this method edge splatting. In order to generate high quality isosurface rendering regardless of the volume resolution and the view, we introduce an edge kernel method. The edge kernel upsamples the isosurface by subdividing every active cell of the volume data. Enough sample points are generated to preserve the exact shape of the isosurface defined by the trilinear interpolation of the volume data. By employing these two methods, we can achieve interactive isosurface exploration with high quality rendering. PMID- 17080862 TI - Using difference intervals for time-varying isosurface visualization. AB - We present a novel approach to out-of-core time-varying isosurface visualization. We attempt to interactively visualize time-varying datasets which are too large to fit into main memory using a technique which is dramatically different from existing algorithms. Inspired by video encoding techniques, we examine the data differences between time steps to extract isosurface information. We exploit span space extraction techniques to retrieve operations necessary to update isosurface geometry from neighboring time steps. Because only the changes between time steps need to be retrieved from disk, I/O bandwidth requirements are minimized. We apply temporal compression to further reduce disk access and employ a point-based previewing technique that is refined in idle interaction cycles. Our experiments on computational simulation data indicate that this method is an extremely viable solution to large time-varying isosurface visualization. Our work advances the state-of-the-art by enabling all isosurfaces to be represented by a compact set of operations. PMID- 17080863 TI - Isosurface extraction and spatial filtering using Persistent OcTree (POT). AB - We propose a novel Persistent OcTree (POT) indexing structure for accelerating isosurface extraction and spatial filtering from volumetric data. This data structure efficiently handles a wide range of visualization problems such as the generation of view-dependent isosurfaces, ray tracing, and isocontour slicing for high dimensional data. POT can be viewed as a hybrid data structure between the interval tree and the Branch-On-Need Octree (BONO) in the sense that it achieves the asymptotic bound of the interval tree for identifying the active cells corresponding to an isosurface and is more efficient than BONO for handling spatial queries. We encode a compact octree for each isovalue. Each such octree contains only the corresponding active cells, in such a way that the combined structure has linear space. The inherent hierarchical structure associated with the active cells enables very fast filtering of the active cells based on spatial constraints. We demonstrate the effectiveness of our approach by performing view dependent isosurfacing on a wide variety of volumetric data sets and 4D isocontour slicing on the time-varying Richtmyer-Meshkov instability dataset. PMID- 17080864 TI - Scalable data servers for large multivariate volume visualization. AB - Volumetric datasets with multiple variables on each voxel over multiple time steps are often complex, especially when considering the exponentially large attribute space formed by the variables in combination with the spatial and temporal dimensions. It is intuitive, practical, and thus often desirable, to interactively select a subset of the data from within that high-dimensional value space for efficient visualization. This approach is straightforward to implement if the dataset is small enough to be stored entirely in-core. However, to handle datasets sized at hundreds of gigabytes and beyond, this simplistic approach becomes infeasible and thus, more sophisticated solutions are needed. In this work, we developed a system that supports efficient visualization of an arbitrary subset, selected by range-queries, of a large multivariate time-varying dataset. By employing specialized data structures and schemes of data distribution, our system can leverage a large number of networked computers as parallel data servers, and guarantees a near optimal load-balance. We demonstrate our system of scalable data servers using two large time-varying simulation datasets. PMID- 17080865 TI - Distributed shared memory for roaming large volumes. AB - We present a cluster-based volume rendering system for roaming very large volumes. This system allows to move a gigabyte-sized probe inside a total volume of several tens or hundreds of gigabytes in real-time. While the size of the probe is limited by the total amount of texture memory on the cluster, the size of the total data set has no theoretical limit. The cluster is used as a distributed graphics processing unit that both aggregates graphics power and graphics memory. A hardware-accelerated volume renderer runs in parallel on the cluster nodes and the final image compositing is implemented using a pipelined sort-last rendering algorithm. Meanwhile, volume bricking and volume paging allow efficient data caching. On each rendering node, a distributed hierarchical cache system implements a global software-based distributed shared memory on the cluster. In case of a cache miss, this system first checks page residency on the other cluster nodes instead of directly accessing local disks. Using two Gigabit Ethernet network interfaces per node, we accelerate data fetching by a factor of 4 compared to directly accessing local disks. The system also implements asynchronous disk access and texture loading, which makes it possible to overlap data loading, volume slicing and rendering for optimal volume roaming. PMID- 17080866 TI - Progressive volume rendering of large unstructured grids. AB - We describe a new progressive technique that allows real-time rendering of extremely large tetrahedral meshes. Our approach uses a client-server architecture to incrementally stream portions of the mesh from a server to a client which refines the quality of the approximate rendering until it converges to a full quality rendering. The results of previous steps are re-used in each subsequent refinement, thus leading to an efficient rendering. Our novel approach keeps very little geometry on the client and works by refining a set of rendered images at each step. Our interactive representation of the dataset is efficient, light-weight, and high quality. We present a framework for the exploration of large datasets stored on a remote server with a thin client that is capable of rendering and managing full quality volume visualizations. PMID- 17080867 TI - Representing higher-order singularities in vector fields on piecewise linear surfaces. AB - Accurately representing higher-order singularities of vector fields defined on piecewise linear surfaces is a non-trivial problem. In this work, we introduce a concise yet complete interpolation scheme of vector fields on arbitrary triangulated surfaces. The scheme enables arbitrary singularities to be represented at vertices. The representation can be considered as a facet-based "encoding" of vector fields on piecewise linear surfaces. The vector field is described in polar coordinates over each facet, with a facet edge being chosen as the reference to define the angle. An integer called the period jump is associated to each edge of the triangulation to remove the ambiguity when interpolating the direction of the vector field between two facets that share an edge. To interpolate the vector field, we first linearly interpolate the angle of rotation of the vectors along the edges of the facet graph. Then. we use a variant of Nielson's side-vertex scheme to interpolate the vector field over the entire surface. With our representation, we remove the bound imposed on the complexity of singularities that a vertex can represent by its connectivity. This bound is a limitation generally exists in vertex-based linear schemes. Furthermore, using our data structure, the index of a vertex of a vector field can be combinatorily determined. We show the simplicity of the interpolation scheme with a GPU-accelerated algorithm for a LIC-based visualization of the so defined vector fields, operating in image space. We demonstrate the algorithm applied to various vector fields on curved surfaces. PMID- 17080868 TI - Techniques for the visualization of topological defect behavior in nematic liquid crystals. AB - We present visualization tools for analyzing molecular simulations of liquid crystal (LC) behavior. The simulation data consists of terabytes of data describing the position and orientation of every molecule in the simulated system over time. Condensed matter physicists study the evolution of topological defects in these data, and our visualization tools focus on that goal. We first convert the discrete simulation data to a sampled version of a continuous second-order tensor field and then use combinations of visualization methods to simultaneously display combinations of contractions of the tensor data, providing an interactive environment for exploring these complicated data. The system, built using AVS, employs colored cutting planes, colored isosurfaces, and colored integral curves to display fields of tensor contractions including Westin's scalar cl, cp, and cs metrics and the principal eigenvector. Our approach has been in active use in the physics lab for over a year. It correctly displays structures already known; it displays the data in a spatially and temporally smoother way than earlier approaches, avoiding confusing grid effects and facilitating the study of multiple time steps; it extends the use of tools developed for visualizing diffusion tensor data, re-interpreting them in the context of molecular simulations; and it has answered long-standing questions regarding the orientation of molecules around defects and the conformational changes of the defects. PMID- 17080869 TI - Diffusion tensor visualization with glyph packing. AB - A common goal of multivariate visualization is to enable data inspection at discrete points, while also illustrating larger-scale continuous structures. In diffusion tensor visualization, glyphs are typically used to meet the first goal, and methods such as texture synthesis or fiber tractography can address the second. We adapt particle systems originally developed for surface modeling and anisotropic mesh generation to enhance the utility of glyph-based tensor visualizations. By carefully distributing glyphs throughout the field (either on a slice, or in the volume) into a dense packing, using potential energy profiles shaped by the local tensor value, we remove undue visual emphasis of the regular sampling grid of the data, and the underlying continuous features become more apparent. The method is demonstrated on a DT-MRI scan of a patient with a brain tumor. PMID- 17080870 TI - Extensions of the Zwart-Powell box spline for volumetric data reconstruction on the cartesian lattice. AB - In this article we propose a box spline and its variants for reconstructing volumetric data sampled on the Cartesian lattice. In particular we present a tri variate box spline reconstruction kernel that is superior to tensor product reconstruction schemes in terms of recovering the proper Cartesian spectrum of the underlying function. This box spline produces a C2 reconstruction that can be considered as a three dimensional extension of the well known Zwart-Powell element in 2D. While its smoothness and approximation power are equivalent to those of the tri-cubic B-spline, we illustrate the superiority of this reconstruction on functions sampled on the Cartesian lattice and contrast it to tensor product B-splines. Our construction is validated through a Fourier domain analysis of the reconstruction behavior of this box spline. Moreover, we present a stable method for evaluation of this box spline by means of a decomposition. Through a convolution, this decomposition reduces the problem to evaluation of a four directional box spline that we previously published in its explicit closed form. PMID- 17080871 TI - A generic and scalable pipeline for GPU tetrahedral grid rendering. AB - Recent advances in algorithms and graphics hardware have opened the possibility to render tetrahedral grids at interactive rates on commodity PCs. This paper extends on this work in that it presents a direct volume rendering method for such grids which supports both current and upcoming graphics hardware architectures, large and deformable grids, as well as different rendering options. At the core of our method is the idea to perform the sampling of tetrahedral elements along the view rays entirely in local barycentric coordinates. Then, sampling requires minimum GPU memory and texture access operations, and it maps efficiently onto a feed-forward pipeline of multiple stages performing computation and geometry construction. We propose to spawn rendered elements from one single vertex. This makes the method amenable to upcoming Direct3D 10 graphics hardware which allows to create geometry on the GPU. By only modifying the algorithm slightly it can be used to render per-pixel iso-surfaces and to perform tetrahedral cell projection. As our method neither requires any pre-processing nor an intermediate grid representation it can efficiently deal with dynamic and large 3D meshes. PMID- 17080872 TI - A spectral analysis of function composition and its implications for sampling in direct volume visualization. AB - In this paper we investigate the effects of function composition in the form g(f(x)) = h(x) by means of a spectral analysis of h. We decompose the spectral description of h(x) into a scalar product of the spectral description of g(x) and a term that solely depends on f(x) and that is independent of g(x). We then use the method of stationary phase to derive the essential maximum frequency of g(f(x)) bounding the main portion of the energy of its spectrum. This limit is the product of the maximum frequency of g(x) and the maximum derivative of f(x). This leads to a proper sampling of the composition h of the two functions g and f. We apply our theoretical results to a fundamental open problem in volume rendering-the proper sampling of the rendering integral after the application of a transfer function. In particular, we demonstrate how the sampling criterion can be incorporated in adaptive ray integration, visualization with multi-dimensional transfer functions, and pre-integrated volume rendering. PMID- 17080873 TI - Improving adhesives. PMID- 17080875 TI - Factors affecting use of fluorescent agents in identification of resin-based polymers. AB - PURPOSE: The present study is a compilation of methodologies developed in order to evaluate the effects of addition of a fluorescent agent, rhodamine B, to resin based materials. The intent of the work was to develop a systematic methodology that accounts for variables not shown to be of concern in past testing, but may significantly affect interpretation of the resulting images and material properties. MATERIALS AND METHODS: Different methodologies were specifically developed to evaluate factors affecting the use of the fluorescent agent rhodamine B in the identification of resin-based materials. The influence of dye concentration was evaluated with respect to adhesive polymerization reaction kinetics and bond strength of the resin into which the dye is added. The pH of the dye-solvent solution was also considered. Additionally, the mass and percentage of rhodamine B leached into different media over time was determined. RESULTS: It was demonstrated that a specific dye must be used with caution, and the concentration of a fluorescent agent is important with respect to both the conversion and bond strength of the resin to which the dye is added. An alcohol based dye solution also produced an extremely high amount of dye leaching from polymerized specimens. The adhesive pH was not altered. CONCLUSION: The standardized methodology developed here to analyze restoration interfaces using a fluorescent dye can lessen the impact of variables when interpreting results. PMID- 17080874 TI - A new technique for assessing hybrid layer interfacial micromorphology and integrity: two-photon laser microscopy. AB - PURPOSE: This study describes a two-photon laser fluorescence microscopy technique developed to evaluate the interfacial micromorphology of the hybrid layer in bonded restorations. MATERIALS AND METHODS: Micropermeability of the hybrid layer was characterized by means of simultaneously contrasting a dye containing adhesive with a differently colored dye placed into the pulp chamber and allowed to diffuse toward the different-colored hybrid layer. A fluorescent red dye (rhodamine B) was incorporated into a commercial dentin bonding agent. Class I preparations (margins in enamel) were made on extracted human third molars. The teeth were restored using conventional methods: bonding agent, composite, finishing, and polishing. An aqueous solution of a yellow/green dye (fluorescein) was then placed into the pulp chamber for 3 h, allowing time to diffuse toward the different-colored bonded interface. The teeth were then embedded, sectioned, and microscopically analyzed using two-photon laser microscopy at 40X magnification. RESULTS: Subsurface fluorescent imaging using this technique enabled interfacial micromorphology to be characterized at submicrometer resolution and provided high-contrast images. The quality of surrounding structures and potential presence of gaps were also precisely assessed. CONCLUSION: Two-photon laser microscopy provided high quality, high resolution images of the bonded interface and surrounding areas, allowing accurate qualitative and quantitative analysis of the structure and integrity of the hybrid layer. PMID- 17080876 TI - Bond strength of resin-based restorations polymerized with different light-curing sources. AB - PURPOSE: To evaluate the influence of different light-curing units on microtensile bond strength of resin composite restorations. MATERIALS AND METHODS: Standardized Class I preparations (6.0 x 4.5 mm, 2.5 mm deep) were made in extracted human third molars after abrading the cusps. Resin was inserted in bulk using a 3M ESPE restorative system [Adper Single Bond (DBA)/ Filtek Z250 (RC)]. Both materials were polymerized using different light-curing units: QTH at 540 mW/cm(2) (XL 3000, 3M ESPE); LED at 750 mW/cm(2) (Elipar FreeLight2, 3M ESPE); PAC at 2130 mW/cm(2) (Arc Light II, Air Techniques). Nine different light combinations were developed to polymerize both DBA and RC: QTH/QTH; QTH/LED; QTH/PAC; LED/LED; LED/QTH; LED/PAC; PAC/PAC; PAC/QTH; PAC/LED. Restored teeth were stored in distilled water for 24 h at 37 degrees C and then sectioned, yielding stick-shaped specimens with a bonded area of approximately 0.9 mm(2). Specimens were assessed in a testing machine at a crosshead speed of 1 mm/min. The results were analyzed using two-way ANOVA and Tukey's test at a pre-set alpha = 0.05. RESULTS: The combinations PAC/QTH and QTH/QTH presented the highest bond strength values, and LED/QTH the lowest (p < 0.05). Significantly lower values were observed in combinations when the LED light was used to polymerize DBA compared to QTH and PAC lights, irrespective of the light source used to polymerize RC (p < 0.05). Same light combinations presented similar bond strength values. CONCLUSIONS: Different light sources influence restoration bond strength. Bond strength is more dependent on the light source used for DBA than for curing RC. PMID- 17080877 TI - Bonding of dual-curing resin cements to dentin. AB - PURPOSE: The purpose of the study was three-fold: 1) to determine the strength of the bond between a number of dual-curing resin cements and dentin treated with corresponding adhesive systems, 2) to determine the effect on bond strength of not light curing the cements, and 3) to investigate whether application of a solution of sodium sulfinate or ascorbic acid would increase the bond strength in the cases where the manufacturer's version of an adhesive system resulted in low bond strength with chemically cured cement, ie, cement cured without light. MATERIALS AND METHODS: The adhesive systems comprised 5 simplified systems (Adper Scotchbond 1 XT, ED Primer II, Excite DSC, OptiBond Solo Plus, and Prime and Bond NT), and as controls, two three-step etch and rinse systems (Adper Scotchbond Multi-Purpose Plus and Gluma Solid Bond). The corresponding dual-curing resin cements were RelyX ARC, Panavia F 2.0, Variolink II, Nexus 2, Calibra, RelyX ARC, and 2Bond2. The cements were either light and chemically cured or only chemically cured. The adhesive systems were used as recommended by the manufacturers, which for some systems involved inclusion of a so-called activator or catalyst when used with chemically cured cement. Sodium sulfinate and ascorbic acid were applied as a 1% ethanol solution. The bond strengths were measured in shear after storing specimens for 24 h in 37 degree C water. RESULTS: When the dual-curing resin cements had been both light and chemically cured, the bond strengths increased in this order: Gluma Solid Bond < ED Primer II < Prime and Bond NT < Adper Scotchbond Multi-Purpose Plus < Excite DSC < Adper Scotchbond 1 XT < OptiBond Solo Plus. Omission of light curing of the cements decreased the bond strengths with OptiBond Solo Plus and Prime and Bond NT. The use of activator in conjunction with OptiBond Solo Plus and Prime and Bond NT increased the bond strength to chemically cured cement, but not to the level obtained when the cement was both light and chemically cured. The use of the catalyst of Adper Scotchbond Multi-Purpose Plus did not increase the bond strength with chemically cured cement. Pretreatment with a solution of sodium sulfinate or ascorbic acid increased the bond strength with chemically cured cement in the case of OptiBond Solo Plus, whereas only the solution of sodium sulfinate was effective with Prime and Bond NT. CONCLUSION: Clinicians should be aware that in situations where a dual-curing resin cement cannot be light cured, some adhesive systems suffer a loss of efficacy, even when respective activators are used. Pretreatment with a 1% ethanol solution of sodium sulfinate may restore much of the lost bond strength. PMID- 17080878 TI - Influence of composite resin consistency and placement technique on proximal contact tightness of Class II restorations. AB - PURPOSE: To investigate the influence of composite resin consistency and placement technique on proximal contact tightness of Class II composite resin restorations. MATERIALS AND METHODS: A manikin model (KaVo Dental) was used with an artificial first molar in which a standardized MO preparation was ground. This preparation was duplicated 360 times. Cavities were restored using Clearfil Photo Bond (Kuraray) combined with one of three composite resins of different consistencies: a low-viscosity (X-Flow, Dentsply), a medium-viscosity (Clearfil AP-X, Kuraray) and a high-viscosity composite (Tetric Ceram HB, Ivoclar Vivadent). Each composite was combined with 6 different matrix systems and separation techniques (n = 20). Groups 1 and 2: precontoured metal circumferential matrix (KerrHawe 1101-c) in a Tofflemire retainer combined either with hand instrument (OptraContact, Ivoclar Vivadent) or separation ring (Composi Tight Gold, Garrison Dental Solutions). Group 3: pre-contoured metal sectional matrix (Lite-Flex, Danville Materials) with separation ring. Groups 4 and 5: pre contoured metal circumferential dead-soft matrix (Adapt SuperCap, KerrHawe) with or without separation ring. Group 6: flat metal circumferential matrix (OptraMatrix, Ivoclar Vivadent) in a Tofflemire-retainer with hand instrument (OptraContact). Proximal contact tightness was measured using the Tooth Pressure Meter (University of Technology, Delft). To determine the effect of experimental variables on the proximal contact tightness, a multiple linear regression model was constructed. RESULTS: Measurements in group 6 were not possible; therefore, this group was excluded. The use of medium- or high-viscosity instead of a low viscosity composite resin resulted in statistically significantly tighter proximal contacts (p < 0.01). The use of a separation ring resulted in a large, statistically significant increase (p < 0.001) in contact tightness, while the use of a hand instrument resulted in a small, statistically significant increase of contact tightness (p = 0.017). No statistically significant differences were found when a dead-soft matrix or a sectional matrix was used instead of a Tofflemire (p = 0.159, p = 0.261, resp.). CONCLUSION: Use of a separation ring when restoring a Class II composite resin restoration has a greater influence on the obtained proximal contact tightness compared to the influence of the consistency of the composite resin. PMID- 17080879 TI - Effects of saliva contamination on dentin bond strength using all-in-one adhesives. AB - PURPOSE: To evaluate the effects of saliva contamination on microtensile bond strengths of all-in-one adhesives to dentin. MATERIALS AND METHODS: Flat superficial occlusal dentin surfaces of extracted human molars were used and randomly divided into 2 groups depending on two all-in-one adhesives: One-Up Bond F Plus (Tokuyama Dental) and Adper Prompt L-Pop (3M ESPE). For each adhesive, the teeth were allocated to 7 subgroups in which one group was bonded without contamination with saliva, serving as a control. The other groups, either before or after the polymerization of adhesives, were contaminated with saliva and dried; contaminated, dried, and adhesives re-applied; contaminated, rinsed, and the adhesives re-applied. Resin composite was then placed on the treated dentin surfaces. After 24 h storage in distilled water at 37 degrees C, the teeth were sectioned perpendicular to the adhesive interface in x and y directions, obtaining multiple beam-shaped specimens with a cross-sectional area of approximately 1.0 mm2. Specimens were stressed in tension at 1 mm/min until rupture of the bond. Data were analyzed using one-way ANOVA and the Games-Howell test. RESULTS: For both all-in-one adhesives, saliva-contaminated groups showed lower bond strengths than the controls. Re-application of the adhesives after drying or rinsing the saliva off restored the dentin bond strengths to the control levels. CONCLUSION: Saliva contamination reduced the dentin bond strengths of all-in-one adhesives. Supplementary application of the adhesive after cleaning the saliva from the dentin surface is an empirical recommendation for restoring bond strength. PMID- 17080880 TI - Shear bond strength of different adhesives to Er:YAG laser-prepared dentin. AB - PURPOSE: The aim of this study was to examine the influence of Er:YAG laser on the shear bond strength of three different adhesives to lased dentin. MATERIALS AND METHODS: Seventy specimens obtained from 35 extracted human molars were embedded in polyester resin and ground with silicon carbide papers. The samples were divided into seven groups. 1. Er:YAG laser (Key Laser 3, KaVo) + Clearfil Protect Bond (Kuraray); 2. Er:YAG laser + Clearfil tri-S Bond (Kuraray); 3. Er:YAG laser + 37% H3PO4 + Single Bond 2 (3M-ESPE); 4. Er:YAG laser + Single Bond 2; 5. conventional method + Clearfil Protect Bond; 6. conventional method + Clearfil tri-S Bond; 7. conventional method + 37% H3PO4 + Single Bond 2. The samples were subjected to shear bond testing 24 h after bonding. Statistical analyses were carried out by two-way ANOVA, t-test, one-way ANOVA, post-hoc Tukey's and Dunnett C test (p = 0.05). RESULTS: Only the Er:YAG laser + Clearfil tri-S Bond group demonstrated significantly higher bond strengths vs conventionally prepared specimens (p < 0.05). There were no significant differences between the shear bond strengths of Single Bond 2 adhesive applied to laser- vs bur-treated specimens (p > 0.05). In laser prepared samples, Clearfil Protect Bond showed the highest scores (p < 0.05), whereas in conventionally prepared groups, no statistical differences were observed between Clearfil Protect Bond and Clearfil tri-S Bond (p > 0.05). CONCLUSION: Er:YAG laser irradiation did not adversely affect the shear bond strength of Single Bond 2 and Clearfil Protect Bond to dentin, whereas it increased the shear bond strength values of Clearfil tri-S Bond. PMID- 17080881 TI - Microtensile bond strength and interfacial properties of self-etching and self adhesive resin cements used to lute composite onlays under different seating forces. AB - PURPOSE: To evaluate strength and morphology of the interface created on enamel and dentin by Panavia F 2.0 (P, Kuraray), RelyX Unicem (RU, 3M ESPE), and Maxcem (M, Sybron-Kerr), applied under two standardized clinically realistic seating pressures. MATERIALS AND METHODS: Composite overlays (Paradigm MZ100, 3M ESPE) were luted on flat enamel or dentin surfaces of 48 extracted molars. During the initial 5-min self-curing period, a pressure of either 20 or 40 g/mm(2) was maintained on the overlay. Curing was completed by light irradiation from the top of the overlay for 20 s. Microtensile sticks and specimens for SEM observations were obtained from the luted teeth. Bond strength data from enamel and dentin (MPa) were analyzed with separate two-way ANOVAs in order to assess the effect of cement type, luting pressure, and their interactions. Tukey's test was used for post-hoc comparisons (alpha = 0.05). RESULTS: The bond strengths (MPa) to dentin were: P 20 g/mm(2) 7.5 +/- 3.7, 40 g/mm(2) 10.9 +/- 4.5; RU 20 g/mm(2) 6.8 +/- 2.6, 40 g/mm(2) 14.5 +/- 5.3; M 20 g/mm(2) 4.1 +/- 1.8, 40 g/mm(2) 5.2 +/- 1.6. The bond strengths (MPa) to enamel were: P 20 g/mm(2) 25.2 +/- 9.0, 40 g/mm(2) 30.7 +/- 8.6; RU 20 g/mm(2) 10.7 +/- 4.9, 40 g/mm(2) 11.1 +/- 5; M 20 g/mm(2) 7.3 +/- 3.1, 40 g/mm(2) 7.9 +/- .2. Cement type, luting pressure, and the interaction of these two factors had significant influences on dentin bond strength, with RU and P outperforming M. RU and P significantly benefited from the increase in luting pressure. In enamel, the type of cement was a significant factor for adhesion, with P yielding the highest and M the lowest strength. The measured bond strengths corresponded with the morphological results. CONCLUSION: Interfacial strength and adaptation of self-etching and self-adhesive dual-curing cements are enhanced if a seating force greater than finger pressure is maintained throughout the initial self-curing period. PMID- 17080882 TI - Microtensile bond strength of self-adhesive luting cements to ceramic. AB - PURPOSE: To test the null hypothesis that three self-adhesive luting materials had the same microtensile bond strength when used to lute ceramic IPS Empress II disks to the dentin of perfused teeth. MATERIALS AND METHODS: Occlusal enamel and the roots of 9 human third molars were removed and crown segments connected to a perfusion system (30 cm H2O). Nine ceramic disks (IPS Empress II) were prepared, conditioned with 5% HF (20 s), rinsed with water, and air dried. A primer silane agent was applied (Monobond-S) for 60 s and air dried. Teeth were bonded to disks using one of three materials: Multilink System, RelyX Unicem, or Panavia F light. Specimens were vertically sectioned to obtain square bars. Each bar was fixed to a rigid custom-made tensile device and submitted to tensile force until debonding. Microtensile bond strength (microTBS) was given in MPa. Because the means of bonded areas (BA) were different between groups, it was impossible to compare microTBS results directly. Thus, the regression line TBS(MPa) = -2.15 + [19.92 / BA (mm(2))] was calculated to correlate microTBS results and BA. Residuals (difference between the value estimated by the regression line and the actual value) were stored as "distances". ANOVA was performed to examine the statistical significance of differences between means distances of the groups', and the Tamhane's post hoc test was used to locate eventual differences. RESULTS: The number of bars used in the analysis was n = 147. Residuals should not be statistically different if the null hypothesis (represented by the regression line) was credible. ANOVA showed statistically significant differences between the distances of the groups' means (p < 0.00001). Tamhane's post hoc test (p < 0.05) showed that different group means were RelyX < Multilink < Panavia F. CONCLUSION: The null hypothesis was rejected: materials showed different tensile bond strengths. Panavia F obtained highest bonding values, followed by Multilink System. RelyX Unicem, the only material that does not require pretreatment of dentin, achieved the lowest TBS values under these study conditions. PMID- 17080884 TI - The placebo effect. PMID- 17080883 TI - Reattachment of a severely traumatized maxillary central incisor, one-year clinical evaluation: a case report. AB - Traumatic fracture of anterior teeth is one of the common dilemmas facing the dentist during daily practice. Classical treatment of a fractured tooth involving the pulp includes the use of post and core with or without a crown, or extraction of the remaining part of the fractured tooth and replacement with a three-unit fixed partial denture or an implant. With the improvement of adhesive systems, reattachment of the fractured tooth fragment became possible as a mode of treatment that immediately improves the esthetic quality of the restored tooth. This article presents a clinical technique for the restoration of a fractured maxillary central incisor, by reattaching the labial enamel fragment using a metallic post and resin composite restoration. One-year clinical evaluation revealed successful results for this technique. PMID- 17080885 TI - Best research for best health: a new national health research strategy. PMID- 17080886 TI - Why choose a career in geriatric medicine? PMID- 17080887 TI - Who's for five nine-hour shifts a week? PMID- 17080888 TI - Care of patients with gastrointestinal disorders. PMID- 17080889 TI - Unexplained (non-cardiac) chest pain. PMID- 17080890 TI - Treating functional lower gastrointestinal symptoms. PMID- 17080891 TI - Colorectal cancer screening. PMID- 17080892 TI - Artificial nutrition support in hospital: indications and complications. PMID- 17080893 TI - The management of upper gastrointestinal haemorrhage. PMID- 17080894 TI - Why do doctors choose a career in geriatric medicine? AB - Reforms to postgraduate training in the U.K. may affect recruitment to geriatric medicine. In 2005, a questionnaire survey was undertaken to determine the factors favouring geriatric medicine as a career choice and whether these might be used to influence recruitment. In all, 1036 responses to the questionnaire were received (response rate 56.4%); 4% of the respondents decided to specialise in geriatric medicine as students, 3.8% of consultants and 8.6% of registrars decided as pre-registration house officers while 39% of consultants and 7% of registrars chose geriatric medicine while a middle grade in another specialty. The strongest influences on choice were clinical aspects of the specialty (34.1%) and inspirational seniors (26.2%). However, 9.2% of consultants and 10.1% of registrars subsequently regretted their career decision. Geriatric medicine seems to be a career choice for doctors of increasing maturity and including more posts in foundation programmes may not improve recruitment as anticipated. Although a small number of doctors regretted choosing geriatric medicine as a career, this was rarely to do with core aspects of the specialty. PMID- 17080895 TI - Choose and Book. AB - Choose and Book combines three major elements of NHS organisational development. Patients are offered increasing choice about where they receive specialist advice, they have more control over the booking of their appointments, and a Choose and Book information technology (IT) application is being implemented to facilitate both these aspects of care. By exploring these three elements, some of the causes of difficulty in clinical implementation and the additional information required to inform choice are identified. PMID- 17080896 TI - Monitoring the introduction of new drugs--Herceptin to cardiotoxicity. AB - Trastuzumab (Herceptin), currently prescribed for metastatic breast cancer, has recently been shown to be effective as adjuvant therapy in early receptor 2 (HER2)-positive breast cancer. Cardiotoxicity is a serious adverse effect. A decrease in left ventricular ejection fraction (LVEF) occurs in as many as 27% of women treated with trastuzumab when combined with standard chemotherapy. The pathophysiology of this effect, which differs from the cardiotoxicity of anthracyclines, remains poorly understood. While overt heart failure is reversed with standard therapy, the longer-term consequences of asymptomatic declines in LVEF remain unknown. Monitoring 3-monthly for 5-10% changes in LVEF, the criteria for cessation of trastuzumab therapy in the clinical trials, is not possible for the population of women who might benefit from trastuzumab for early breast cancer. Extension of this therapy to an older and less fit population than those enrolled in the trials, with less rigorous cardiac screening, may result in significantly more cardiotoxicity. PMID- 17080897 TI - The elusive goal of liver support--quest for the Holy Grail. AB - The history of liver support devices is traced from early attempts with dialysis based on the known dialysability of ammonia--the major identified toxin in liver failure--and exchange transfusion with removal of protein-bound toxins, to the later techniques based on whole organ perfusion in extracorporeal circuits. Perfusion through charcoal as an adsorbent represented a major advance and remains a component of more recently introduced devices based on bioreactors of cultured hepatocytes and in the albumin dialysis techniques of molecular adsorbent recirculating system and the Prometheus device. The latter are the most highly efficient to date in toxin removal but whether survival is improved and the need for liver transplantation remain to be proven. PMID- 17080898 TI - Leadership in academic medicine: reflections from administrative exile. AB - Physicians are drawn into diverse leadership roles in academic medicine, but little in our education and training prepares us for these responsibilities. Fortunately, there is growing convergence in the literature on the attributes of successful leaders for knowledge-based organisations. Top-performing leaders seem to be self-effacing team-builders who eschew rapid-cycle strategic planning and management trends, focusing instead on strategic and incremental changes that will gradually transform their organisations. Academic physicians and search committees often concentrate on personal achievement and intellectual or technical mastery in research and clinical care. In contrast, the literature on leadership suggests other-directed skills matter more, eg mentorship, learning and teaching competencies, and so-called emotional intelligence. As a corollary, teaching hospitals, universities, and professional colleges or societies are long term organisations with a rich history. Leadership in such a context demands stewardship of tradition along with patient pursuit of changes required to ensure that the organisation evolves successfully. PMID- 17080899 TI - U.K. health challenges: can the E.U. make a difference? PMID- 17080900 TI - Transition: getting it right for young people. PMID- 17080901 TI - Mother and child care in art. The captured truant. PMID- 17080902 TI - Rationing of medical care by age. PMID- 17080903 TI - Systematic review of systematic reviews of acupuncture. PMID- 17080904 TI - Assisted dying. PMID- 17080905 TI - Assisted dying. PMID- 17080906 TI - Delay in the presentation of patients to hospital is the main factor for failure to adhere to the Royal College of Physicians' guidelines in the management of suspected subarachnoid haemorrhage. PMID- 17080908 TI - Clinical examination in three minutes? PMID- 17080907 TI - HIV testing for adult patients with Streptococcus pneumoniae bacteraemia. PMID- 17080909 TI - Characterization of T-cell population in children with prolonged fetal exposure to dexamethasone for anti-Ro/SS-A antibodies associated congenital heart block. AB - The objectives of the study were to characterize the production, function and survival of T lymphocytes of children with prolonged fetal exposure to dexamethasone for anti-Ro/SS-A antibodies associated congenital complete heart block. The analysis of thymic function, studied by measuring the level of T-cell receptor excision circles, was performed by real time PCR, the composition of T cell subpopulation was evaluated by flow cytometry and the T-cell diversity was assayed by heteroduplex analysis. T-cell competence was gauged at two functional levels by determining the proliferation and the number of T-cell divisions and by measuring gamma-interferon production after mitogenic stimulation. We observed that the thymic output, distribution of T-cell subsets, thymidine incorporation, number of T-cell divisions, and y-interferon production were comparable to those of age-matched control. On the contrary, heteroduplex analysis demonstrated the presence of both polyclonal and oligoclonal peripheral T-cell repertoires. In conclusion, the analysis of the T-cell compartment in children with prolonged intrauterine exposure to high dose dexamethasone did not disclose any relevant abnormality, except a restriction of T-cell receptor diversity in some patients. PMID- 17080910 TI - Cardiovascular risk scores and the presence of subclinical coronary artery atherosclerosis in women with systemic lupus erythematosus. AB - The Framingham risk score is widely used to identify patients at increased cardiovascular risk, and women with systemic lupus erythematosus (SLE) have a marked increased prevalence of cardiovascular events. Thus, we examined the hypothesis that cardiovascular risk scores would identify women with SLE who had asymptomatic coronary atherosclerosis. Ninety-three women with SLE and 65 control subjects were studied. The Framingham score and a score for younger populations developed from the Pathobiological Determinants of Atherosclerosis in Youth (PDAY) study were compared in both groups. Coronary atherosclerosis was ascertained by electron beam computed tomography. There were no significant differences in the median (interquartile range) Framingham [5 (2-10) compared to 7 (0-10), P = 0.88] and PDAY [15 (14-18) compared to 16 (13-18), P = 0.99] scores in patients with SLE and controls, respectively. Coronary atherosclerosis was associated with higher Framingham [12 (3-15) compared to 4 (1-8), P = 0.008] and PDAY [17 (15-19 compared to 15 (12-18), P = 0.03)] scores in patients with SLE; however, 99% of patients were classified as low-risk with a 10-year predicted risk of 1% (<1-3%). Our data indicate that cardiovascular risk scores are not adequate for risk stratification in women with SLE. Measurement of coronary calcification may add information to identify asymptomatic women with lupus who might benefit from aggressive preventive measures. PMID- 17080911 TI - B-lymphocyte activating factor in systemic lupus erythematosus and rheumatoid arthritis in relation to autoantibody levels, disease measures and time. AB - Overexpression of B-lymphocyte activating factor (BAFF) results in arthritis, glomerulonephritis and autoantibody formation in mice, but its role in human autoimmune disease is less obvious. Serum BAFF levels in patients with systemic lupus erythematosus (SLE) (n=42) and rheumatoid arthritis (RA) (n=60) were related to levels of disease activity, anti-dsDNA Ab, anti-ENA Ab, rheumatoid factor (RF) and anti-CCP Ab. BAFF levels were also followed over time in 19 SLE patients. BAFF levels correlated inversely with age, were higher in SLE than RA (median 2.7 versus 1.4 ng/mL, P < 0.01) and more SLE than RA patients had increased BAFF levels (57% versus 10%, P < or = 0.01). In SLE, BAFF levels correlated with SLEDAI scores but not with anti-dsDNA Ab levels. SLE patients with increased BAFF levels had higher SLEDAI and CRP levels. In RA, BAFF levels correlated weakly with anti-CCP levels (Rs 0.27, P = 0.07), but not with joint counts, ESR, CRP or RF levels. Longitudinal BAFF levels remained unaltered in two thirds of SLE patients and changes in BAFF levels were unrelated to disease flares. These findings suggest that BAFF stimulation of B-cells may contribute to SLE by other mechanisms than autoantibody production. PMID- 17080912 TI - Effect of antimalarials on thrombosis and survival in patients with systemic lupus erythematosus. AB - Antimalarials have shown beneficial effects on systemic lupus erythematosus (SLE) activity. Our aim was to investigate whether antimalarials protect against thrombosis and influence survival in SLE patients. A prospective cohort including 232 patients with SLE were included in the study at the time of lupus diagnosis. End points were documented thrombosis and death due to any cause. A Cox regression-multiple-failure time survival analysis model was fitted to establish the effect of antimalarials on the development of thrombosis. Kaplan-Meier survival curves and propensity score adjusted-Cox regression analysis were performed to investigate the effect of antimalarials use on survival. Of our subjects, 204 patients (88%) were women. 230 patients (99%) were white. 150 patients (64%) had ever received antimalarials. Median time on antimalarials was 52 months (range three to 228 months). The Cox multiple-failure time survival analysis showed that taking antimalarials was protective against thrombosis (HR 0.28, 95% CI 0.08-0.90), while aPL-positivity (HR 3.16, 95% CI 1.45-6.88) and previous thrombosis (HR 3.85, 95% CI 1.50-9.91) increased the risk of thrombotic events. Twenty-three patients died, 19 of whom (83%) had never received antimalarials. No patient treated with antimalarials died of cardiovascular complications. Cumulative 15-year survival rates were 0.68 for never versus 0.95 for ever treated patients (P < 0.001). Age at diagnosis and propensity score adjusted HR for antimalarials ever versus never users was 0.14 (95% CI 0.04 0.48). Our study shows a protective effect of antimalarials against thrombosis and an increased survival of SLE patients taking these drugs. These data support the routine use of antimalarials in all patients with SLE. PMID- 17080913 TI - Infections in systemic lupus erythematosus: a prospective and controlled study of 110 patients. AB - We decided to analyse the incidence and characteristics of infection in systemic lupus erythematosus (SLE) and determine the related risks factors. One-hundred and ten SLE patients and 220 controls were prospectively followed up over three years and all the infectious episodes were recorded. A case-control design was established to identify risk factors of infection. Thirty-nine SLE patients suffered at least one infection (36%) versus 53 controls (22%), RR = 1.63 (P < 0.05). The incidence of urinary infections, pneumonia and bacteraemia without known focus was significantly greater in SLE. E. coli was the chief isolated microorganism (21.3%). In the univariate analysis, nephritis, SLE activity, leucopenia, anti-dsDNA levels above 20 IU/mL, CH50 levels under 300 IU/mL, ever use of steroids, daily dose of prednisone higher than 10 mg and ever use of cyclophosphamide were significantly associated with infection. In the multivariate analysis, total serum complement levels below 300 UU/mL and a daily dose of prednisone above 20 mg during at least one month plus ever use of cyclophosphamide were found to be significant (P < 0.0001). We conclude that patients with SLE have an increased overall risk for infection and they are especially prone to develop urinary infection, pneumonia and bacteraemia without focus. Hypocomplementaemia represents an independent predictive factor for infection. It seems mandatory to closely follow up SLE patients with low complement levels and instruct them to report any suspicious sign of infection, especially in those receiving more than 20 mg/day of prednisone who have also been administered cyclophosphamide. PMID- 17080914 TI - A comparison of damage accrual across different calendar periods in systemic lupus erythematosus patients. AB - Therapeutic approaches in systemic lupus erythematosus (SLE) have evolved over the last few decades, but their impact on prevention of organ damage is unknown. The objective of this study was to compare new cumulative damage in SLE patients across different calendar periods. Patients from a large SLE cohort were divided into two subcohorts; the first diagnosed and followed between 1978 and 1988 (cohort #1, n=100) and the second between 1989 and 1999 (cohort #2, n=51). Initial Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index (SLICC/ACR DI) scores, and changes in scores over the observation intervals, were compared for the two groups. Logistic regression estimated adjusted odds ratios (OR) comparing damage accrual between the two cohorts. Medication exposures were noted. Baseline characteristics were similar between the two groups. At first assessment, the adjusted OR for a SLICC/ACR DI score > or =1 was 1.79 (95% CI 0.82, 3.88) for cohort #1 versus cohort #2. At the end of the observation interval, the adjusted OR for a SLICC/ACR DI score > or =1 was 1.22 (0.58, 2.55) for cohort #1 versus cohort #2. The adjusted OR for accruing damage over the observation interval in cohort #1 versus cohort #2 was 0.94 (0.39, 2.44). Increased medication exposure was evident for cohort #2 compared to cohort #1. Despite increased therapeutic measures used for patients in more recent periods, our data do not establish a clear difference in damage accrual. This emphasizes the need for strategies to effectively treat lupus specific manifestations, while minimizing side effects and comorbidities. PMID- 17080915 TI - Outcome of pregnancies complicated by systemic sclerosis and mixed connective tissue disease. AB - Systemic sclerosis (SSc) and mixed connective tissue disease (MCTD) are rare autoimmune diseases which share the common feature of non-inflammatory vasculopathy. Studies evaluating pregnancy outcomes in these patients have yielded conflicting results. We sought to describe the outcomes of pregnancies associated with SSc and MCTD followed at our center utilizing a retrospective review of all pregnant women with SSc and MCTD followed at Stanford University from 1993 to 2003. We identified 20 pregnancies occurring in 13 women with SSc or MCTD. Twelve pregnancies occurred in seven women with SSc and eight pregnancies occurred in six women with MCTD. The overall preterm delivery rate was 39% and small for gestational age infants occurred in 50% and 63% of pregnancies associated with SSc and MCTD, respectively. Fetal loss complicated two pregnancies in women with severe diffuse SSc and the antiphospholipid antibody syndrome. There were no cases of congenital heartblock among infants, and only one case of pre-eclampsia was observed. Maternal flares of disease during pregnancy were generally mild. Most pregnancies in women with SSc and MCTD in this cohort were uncomplicated. The high rates of prematurity and small for gestational age infants underscore the risk for growth restriction consistent with the vasculopathy associated with these diseases. PMID- 17080916 TI - Diminished expression of complement regulatory proteins (CD55 and CD59) in lymphocytes from systemic lupus erythematosus patients with lymphopenia. AB - CD55 and CD59 are glycophosphatidylinositol-anchored proteins with complement inhibitory properties. Lymphopenia in systemic lupus erythematosus (SLE) has been associated with autoantibodies targeting nuclear antigens. The aim of this study was to evaluate the surface density of CD55 and CD59 in T and B lymphocytes from patients with SLE and lymphopenia and its possible correlation with the presence of common SLE autoantibodies. Flow cytometric analyses were performed on CD55 and CD59 stained CD3+ and CD19+ cells from 40 SLE patients, 30 with lymphopenia and 10 without it, and 25 healthy controls. Autoantibodies were detected in the sera by enzyme linked immunosorbent assay. The mean fluorescence intensity of CD55 and CD59 in T and B cells was significantly diminished in SLE patients with lymphopenia when compared with healthy subjects. Interestingly, the opposite was found in T and B cells from non-lymphopenic SLE patients. Although there was no correlation between CD55 and CD59 surface density and the presence of any specificity of the autoantibodies tested, higher titres of anti-dsDNA, anti-SM and anti-ribosomal p antibodies were significantly associated with lymphopenia. The deficiency of CD55 and CD59 expression may play a role in the pathophysiology of lymphopenia, most likely by increasing the susceptibility of cells to complement mediated cytolysis. PMID- 17080917 TI - Systemic Lupus International Collaborating Group--onwards and upwards? PMID- 17080918 TI - Transforming growth factor beta 1 in children with systemic lupus erythematosus: a possible relation with clinical presentation of lupus nephritis. AB - Plasma and urinary (latent and active) TGF-beta1 levels were assessed in 32 children with active lupus and compared to 15 healthy controls of matched age and sex. Plasma latent and active TGF-beta1 levels in children with active disease were significantly lower than controls (P = 0.004 and P < 0.001 respectively). Plasma active TGF-beta1 correlated negatively with Systemic Lupus Erythematosus Disease Activity Index (r = -0.38, P = 0.03). On the contrary, urinary latent and active TGF-beta1 levels in children with active disease were significantly higher than controls (P < 0.001 and P = 0.003 respectively). Urinary active TGF-beta1 levels correlated positively with Anti-ds DNA titre (r = 0.42, P = 0.015) and negatively with serum C3 levels (r = -0.48, P = 0.005). Patients with symptomatic nephritis had significantly elevated urinary active TGF-beta1 levels in comparison to those with silent nephritis (P = 0.008). From this data we conclude that lowered plasma TGF-beta1 levels may be a feature of systemic immune dysfunction in children with active lupus while increased renal production of active TGF-beta1 seems to have a role in the clinical presentation of lupus nephritis. PMID- 17080919 TI - Renal MALToma: an unusual lymphoma in a patient with lupus. AB - Non-Hodgkin's Lymphomas (NHL) have been reported in association with autoimmune disorders particularly Sjogren's syndrome. We report a case of renal MALToma, an unusual NHL in an 84-year-old Caucasian lady with long-standing, non-aggressive Systemic Lupus Erythematosis with no associated Sjorgen's syndrome and who never received cytotoxics. PMID- 17080920 TI - Juvenile systemic lupus erythematosus in multicase families from Saudi Arabia: comparison of clinical and laboratory variables with sporadic cases. AB - The object of this study was to compare patients with familial versus sporadic systemic lupus erythematosus (SLE) with respect to clinical, laboratory variables and outcome. The familial SLE group comprised 12 patients while the comparative group comprised 24 patients selected by systemic sampling from our pediatric rheumatology clinic database. Those patients are listed according to the date of referral, which represents a sampling frame. The first patient was chosen randomly and subsequent patients were chosen at intervals of three. The two groups were compared with respect to: demographic information, age of onset of SLE, disease and follow up duration, clinical and laboratory variables and outcome. The patients from the familial group were younger and had an earlier age of onset of disease (P = 0.03, 0.001 respectively). Seven patients with familial SLE were from the eastern region of Saudi Arabia (P = 0.006). The two groups were comparable with respect to gender, disease duration and follow-up. At diagnosis, the discoid rash was more frequent in the familial group (P = 0.03) while other clinical and laboratory variables including disease activity as measured by SLEDAI did not show significant differences. The mean dose of steroid and use of other immunosuppressive therapy were similar in both groups. Three patients from the familial group died; two of them had unusual complications (one patient had transverse myelitis and pancreatic pseudocyst and the other one had extensive pyoderma gangrenosum). All patients from the sporadic group are alive in stable condition but one patient had severe central nervous system disease. Familial SLE patients tend to be younger and more likely to have discoid rash, in addition a marked difference in the origin of patients was noted. These differences may be helpful in identifying SLE patients with a stronger genetic predisposition. The mortality among familial SLE patients is more frequent which may reflect the disease severity. PMID- 17080921 TI - Paraneoplastic dermatomyositis associated with bladder cancer. PMID- 17080923 TI - Requiring 'independent' statistical analyses for industry sponsored trials? PMID- 17080922 TI - Systemic lupus erythematosus in a multiethnic cohort (LUMINAXXXIX): relationship between hormone replacement therapy and disease activity over time. PMID- 17080924 TI - Review of guidelines and literature for handling missing data in longitudinal clinical trials with a case study. AB - Missing data in clinical trials are inevitable. We highlight the ICH guidelines and CPMP points to consider on missing data. Specifically, we outline how we should consider missing data issues when designing, planning and conducting studies to minimize missing data impact. We also go beyond the coverage of the above two documents, provide a more detailed review of the basic concepts of missing data and frequently used terminologies, and examples of the typical missing data mechanism, and discuss technical details and literature for several frequently used statistical methods and associated software. Finally, we provide a case study where the principles outlined in this paper are applied to one clinical program at protocol design, data analysis plan and other stages of a clinical trial. PMID- 17080925 TI - Gatekeeping procedures in dose-response clinical trials based on the Dunnett test. AB - This paper discusses multiple testing procedures in dose-response clinical trials with primary and secondary endpoints. A general gatekeeping framework for constructing multiple tests is proposed, which extends the Dunnett test [Journal of the American Statistical Association 1955; 50: 1096-1121] and Bonferroni-based gatekeeping tests developed by Dmitrienko et al. [Statistics in Medicine 2003; 22:2387-2400]. The proposed procedure accounts for the hierarchical structure of the testing problem; for example, it restricts testing of secondary endpoints to the doses for which the primary endpoint is significant. The multiple testing approach is illustrated using a dose-response clinical trial in patients with diabetes. Monte-Carlo simulations demonstrate that the proposed procedure provides a power advantage over the Bonferroni gatekeeping procedure. The power gain generally increases with increasing correlation among the endpoints, especially when all primary dose-control comparisons are significant. PMID- 17080926 TI - Sample size calculations for clinical studies allowing for uncertainty about the variance. AB - One of the most important steps in the design of a pharmaceutical clinical trial is the estimation of the sample size. For a superiority trial the sample size formula (to achieve a stated power) would be based on a given clinically meaningful difference and a value for the population variance. The formula is typically used as though this population variance is known whereas in reality it is unknown and is replaced by an estimate with its associated uncertainty. The variance estimate would be derived from an earlier similarly designed study (or an overall estimate from several previous studies) and its precision would depend on its degrees of freedom. This paper provides a solution for the calculation of sample sizes that allows for the imprecision in the estimate of the sample variance and shows how traditional formulae give sample sizes that are too small since they do not allow for this uncertainty with the deficiency being more acute with fewer degrees of freedom. It is recommended that the methodology described in this paper should be used when the sample variance has less than 200 degrees of freedom. PMID- 17080927 TI - Implementation of a Bayesian adaptive design in a proof of concept study. AB - With increased costs of drug development the need for efficient studies has become critical. A key decision point on the development pathway has become the proof of concept study. These studies must provide clear information to the project teams to enable decision making about further developing a drug candidate but also to gain evidence that any effect size is sufficient to warrant this development given the current market environment. Our case study outlines one such proof of concept trial where a new candidate therapy for neuropathic pain was investigated to assess dose-response and to evaluate the magnitude of its effect compared to placebo. A Normal Dynamic Linear Model was used to estimate the dose-response--enforcing some smoothness in the dose-response, but allowing for the fact that the dose-response may be non-monotonic. A pragmatic, parallel group study design was used with interim analyses scheduled to allow the sponsor to drop ineffective doses or to stop the study. Simulations were performed to assess the operating characteristics of the study design. The study results are presented. Significant cost savings were made when it transpired that the new candidate drug did not show superior efficacy when compared placebo and the study was stopped. PMID- 17080928 TI - Simple graphical methods of displaying multiple clinical results. AB - A common challenge in clinical research trials is for applied statistics to manage, analyse, summarize and report an enormous amount of data. Nowadays, due to advances in medical technology, situations frequently arise where it is difficult to display and interpret results. Consequently, a creative approach is required to summarize the main outcomes of the statistical analyses in a form which is easy to grasp, to interpret and possibly to remember. In this paper a number of clinical case studies are provided. Firstly, a topographical map of the brain summarizing P-values obtained from comparisons across different EEG sites; secondly, a bulls eye plot, showing the agreement between observers in different regions of the heart; thirdly, a pictorial table reporting inter- and intra-rater reliability scores of a speech assessment; fourthly a star-plot to deal with numerous questionnaire results and finally a correlogram to illustrate significant correlation values between two diagnostic tools. The intention of this paper is to encourage the effort of visual representations of multiple statistical outcomes. Such representations do not only embellish the report, but aid interpretation by conveying a specific statistical meaning. PMID- 17080929 TI - Adaptive designs for pivotal trials: discussion points from the PSI Adaptive Design Expert Group. AB - The Committee for Medicinal Products for Human Use (CHMP) is currently preparing a guideline on 'methodological issues in confirmatory clinical trials with flexible design and analysis plan'. PSI (Statisticians in the Pharmaceutical Industry) sponsored a meeting of pharmaceutical statisticians with an interest in the area to share experiences and identify potential opportunities for adaptive designs in late-phase clinical drug development. This article outlines the issues raised, resulting discussions and consensus views reached. Adaptive designs have potential utility in late-phase clinical development. Sample size re-estimation seems to be valuable and widely accepted, but should be made independent of the observed treatment effect where possible. Where unblinding is necessary, careful consideration needs to be given to preserving the integrity of the trial. An area where adaptive designs can be particularly beneficial is to allow dose selection in pivotal trials via adding/dropping treatment arms; for example, combining phase II and III of the drug development program. The more adaptations made during a late-phase clinical trial, the less likely that the clinical trial would be considered as a confirmatory trial. In all cases it would be advisable to consult with regulatory agencies at the protocol design stage. All involved should remain open to scientifically valid opportunities to improve drug development. PMID- 17080930 TI - Water relations of baobab trees (Adansonia spp. L.) during the rainy season: does stem water buffer daily water deficits? AB - Baobab trees are often cited in the literature as water-storing trees, yet few studies have examined this assumption. We assessed the role of stored water in buffering daily water deficits in two species of baobabs (Adansonia rubrostipa Jum. and H. Perrier and Adansonia za Baill.) in a tropical dry forest in Madagascar. We found no lag in the daily onset of sap flow between the base and the crown of the tree. Some night-time sap flow occurred, but this was more consistent with a pattern of seasonal stem water replenishment than with diurnal usage. Intrinsic capacitance of both leaf and stem tissue (0.07-0.08 and 1.1-1.43 MPa(-1), respectively) was high, yet the amount of water that could be withdrawn before turgor loss was small because midday leaf and stem water potentials (WPs) were near the turgor-loss points. Stomatal conductance was high in the daytime but then declined rapidly, suggesting an embolism-avoidance strategy. Although the xylem of distal branches was relatively vulnerable to cavitation (P50: 1.1 1.7 MPa), tight stomatal control and minimum WPs near--1.0 MPa maintained native embolism levels at 30-65%. Stem morphology and anatomy restrict water movement between storage tissues and the conductive pathway, making stored-water usage more appropriate to longer-term water deficits than as a buffer against daily water deficits. PMID- 17080931 TI - Over-expression of different aldehyde dehydrogenase genes in Arabidopsis thaliana confers tolerance to abiotic stress and protects plants against lipid peroxidation and oxidative stress. AB - Aldehyde dehydrogenases (ALDHs) play a major role in the detoxification processes of aldehydes generated in plants when exposed to abiotic stress. In previous studies, we have shown that the Arabidopsis thaliana ALDH3I1 gene is transcriptionally activated by abiotic stress, and over-expression of the ALDH3I1 gene confers stress tolerance in transgenic plants. The A. thaliana genome contains 14 ALDH genes expressed in different sub-cellular compartments and are presumably involved in different reactions. The purpose of this study was to compare the potential of a cytoplasmic and a chloroplastic stress-inducible ALDH in conferring stress tolerance under different conditions. We demonstrated that constitutive or stress-inducible expression of both the chloroplastic ALDH3I1 and the cytoplasmic ALDH7B4 confers tolerance to osmotic and oxidative stress. Stress tolerance in transgenic plants is accompanied by a reduction of H2O2 and malondialdehyde (MDA) derived from cellular lipid peroxidation. Involvement of ALDHs in stress tolerance was corroborated by the analysis of ALDH3I1 and ALDH7B4 T-DNA knockout (KO) mutants. Both mutant lines exhibited higher sensitivity to dehydration and salt than wild-type (WT) plants. The results indicate that ALDH3I1 and ALDH7B4 not only function as aldehyde-detoxifying enzymes, but also as efficient reactive oxygen species (ROS) scavengers and lipid peroxidation inhibiting enzymes. The potential of ALDHs to interfere with H2O2 was also shown for recombinant bacterial proteins. PMID- 17080932 TI - Senescence-specific regulation of catalases in Arabidopsis thaliana (L.) Heynh. AB - Oxygen free radicals are thought to play an essential role in senescence, especially those derived from peroxisomes. Therefore, the activities of different isoforms of the peroxisomal hydrogen peroxide (H2O2)-scavenging enzyme catalase (CAT) were analysed during senescence of Arabidopsis. CAT2 activity decreased with bolting time parallel with cytosolic ascorbate peroxidase 1 (APX1) activity before loss of chlorophyll could be measured. At the same time point, the H2O2 content increased. Subsequently, the stress-inducible CAT3 isoform was activated and APX1 activity was recovered, accompanied by a decline of the H2O2 content. In very late stages, low activities of the seed-specific CAT1 became detectable in leaves, but H2O2 increased again. Further analyses of CAT expression by promoter: beta-glucuronidase (GUS) fusions in transgenic plants revealed a vasculature specific CAT3 expression, whereas CAT2 expression turned out to be specific for photosynthetic active tissues. CAT2 expression is down-regulated during leaf senescence, while CAT3 expression is induced with age and corresponds to an accumulation of H2O2 in the vascular bundles. CAT2 down-regulation on the transcriptional level appears as the initial step in creating the H2O2 peak during bolting time, while the decrease in APX1 activity might only be a secondary and amplifying effect. PMID- 17080933 TI - Metabolic consequences of susceptibility and resistance (race-specific and broad spectrum) in barley leaves challenged with powdery mildew. AB - In a compatible interaction biotrophic fungi often lower the yield of their hosts by reducing photosynthesis and altering the fluxes of carbon within the infected leaf. In contrast, comparatively little is known about the metabolic consequences of activating resistance responses. In this study we investigated the hypothesis that the activation of both race-specific (Mla12) and broad-spectrum (mlo) resistance pathways in barley leaves infected with Blumeria graminis represents a cost to the plant in terms of carbon production and utilization. We have shown, using quantitative imaging of chlorophyll fluorescence, that during a susceptible interaction, photosynthesis was progressively reduced both in cells directly below fungal colonies and in adjacent cells when compared with uninoculated leaves. The lower rate of photosynthesis was associated with an increase in invertase activity, an accumulation of hexoses and a down-regulation of photosynthetic gene expression. During both Mla12- and mlo-mediated resistance, photosynthesis was also reduced, most severely inhibited in cells directly associated with attempted penetration of the fungus but also in surrounding cells. These cells displayed intense autofluorescence under ultraviolet illumination indicative of the accumulation of phenolic compounds and/or callose deposition. The depression in photosynthesis was not due only to cell death but also to an alteration in source-sink relations and carbon utilization. Apoplastic (cell wall-bound) invertase activity increased more rapidly and to a much greater extent than in infected susceptible leaves and was accompanied by an accumulation of hexoses that was localized to areas of the leaf actively exhibiting resistance responses. The accumulation of hexoses was accompanied by a down-regulation in the expression of Rubisco (rbcS) and chlorophyll a/b binding protein (cab) genes (although to a lesser extent than in a compatible interaction) and with an up regulation in the expression of the pathogenesis-related protein 1 (PR-1). These results are consistent with a role for invertase in the generation of hexoses, which may supply energy for defence reactions and/or act as signals inducing defence gene expression. PMID- 17080934 TI - Higher daytime leaf temperatures contribute to lower freeze tolerance under elevated CO2. AB - Elevated atmospheric CO2 adversely affects freezing tolerance in many evergreens, but the underlying mechanism(s) have been elusive. We compared effects of elevated CO2 with those of daytime warming on acclimation of snow gum (Eucalyptus pauciflora) to freezing temperatures under field conditions. Reduction in stomatal conductance g(c) under elevated CO2 was shown to cause leaf temperature to increase by up to 3 degrees C. In this study, this increase in leaf temperature was simulated under ambient CO2 conditions by using a free air temperature increase (FATI) system to warm snow gum leaves during daytime, thereby increasing the diurnal range in temperature without affecting temperature minima. Acclimation to freezing temperatures was assessed using measures of electrolyte leakage and photosynthetic efficiency of leaf discs exposed to different nadir temperatures. Here, we show that both elevated CO2 and daytime warming delayed acclimation to freezing temperatures for 2-3 weeks after which time freeze tolerance of the treated plants in both the FATI and open top chamber (OTC) experiments did not differ from control plants. Our results support the hypothesis that delayed development of freezing tolerance under elevated CO2 is because of higher daytime leaf temperatures under elevated CO2. Thus, potential gains in productivity in response to increasing atmospheric CO2 and prolonging the growing season may be reduced by an increase in freezing stress in frost prone area. PMID- 17080935 TI - Using a standard framework for the phenotypic analysis of Medicago truncatula: an effective method for characterizing the plant material used for functional genomics approaches. AB - A crucial step for identifying genes of interest in legume crops is to determine gene function in Medicago truncatula. To facilitate functional genomics in this species, an ecophysiological framework of analysis was developed. Our primary aim was to establish a standard terminology for identifying each organ on the plant. A standard system for the characterization of the vegetative and the reproductive developmental stages was then proposed. Using these tools, the time course of vegetative development of nitrogen-fixing A17 plants was analysed in experiments conducted under different environmental conditions. To take into account the influence of temperature on plant development timing, an original approach was used by modelling vegetative development as a function of thermal time. Interestingly, the use of thermal time highlighted genotypic constants in plant development. Thereafter, to illustrate how this methodology can be used in explaining phenotypic alterations, the phenotype of two allelic mutants was analysed. Because the tools proposed in this paper allow the following: (1) standardization of how the plant material should be characterized to be used for functional genomics; (2) prediction of plant vegetative development; and (3) a more accurate phenotyping, the use of these tools by the M. truncatula community should provide a relevant framework for facilitating the production of reproducible functional genomics data. PMID- 17080936 TI - Shade avoidance and the regulation of leaf inclination in Arabidopsis. AB - As a rosette plant, Arabidopsis thaliana forms leaves near to the ground, which causes the plant to be vulnerable to shading by neighbours. One mechanism to avoid such shading is the regulation of leaf inclination, such that leaves can be raised to more vertical orientations to prevent neighbouring leaves from overtopping them. Throughout Arabidopsis rosette development, rosette leaves move to more vertical orientations when shaded by neighbouring leaves, exposed to low light levels or placed in the dark. After dark-induced reorientation of leaves, returning them to white light causes the leaves to reorient to more horizontal inclinations. These light-dependent leaf movements are more robust than, and distinct from, the diurnal movements of rosette leaves. However, the movements are gated by the circadian clock. The light-dependent leaf orientation response is mediated primarily through phytochromes A, B and E, with the orientation varying with the ratio of red light to far-red light, consistent with other shade avoidance responses. However, even plants lacking these phytochromes were able to alter leaf inclination in response to white light, suggesting a role for other photoreceptors. In particular, we found significant changes in leaf inclination for plants exposed to green light. This green light response may be caused, in part, by light-dependent regulation of abscisic acid (ABA) biosynthesis. PMID- 17080937 TI - Microelectrode ion and O2 fluxes measurements reveal differential sensitivity of barley root tissues to hypoxia. AB - Hypoxia-induced changes in net H+, K+ and O2 fluxes across the plasma membrane (PM) of epidermal root cells were measured using the non-invasive microelectrode ion flux measurement (MIFE) system in elongation, meristem and mature root zones of two barley (Hordeum vulgare L.) varieties contrasting in their waterlogging (WL) tolerance. The ultimate goal of this study was to shed light on the mechanisms underlying effects of WL on plant nutrient acquisition and mechanisms of WL tolerance in barley. Our measurements revealed that functionally different barley root zones have rather different O2 requirements, with the highest O2 influx being in the elongation zone of the root at about 1 mm from the tip. Oxygen deprivation has qualitatively different effects on the activity of PM ion transporters in mature and elongation zones. In the mature zone, hypoxic treatment caused a very sharp decline in K+ uptake in the WL sensitive variety Naso Nijo, but did not reduce K+ influx in the WL tolerant TX9425 variety. In the elongation zone, onset of hypoxia enhanced K+ uptake from roots of both cultivars. Pharmacological experiments suggested that hypoxia-induced K+ flux responses are likely to be mediated by both K(+) -inward- (KIR) and non-selective cation channels (NSCC) in the elongation zone, while in the mature zone K(+) outward- (KOR) channels are the key contributors. Overall, our results suggest that oxygen deprivation has an immediate and substantial effect on root ion flux patterns, and that this effect is different in WL-sensitive and WL-tolerant cultivars. To what extent this difference in ion flux response to hypoxia is a factor conferring WL tolerance in barley remains to be answered in future studies. PMID- 17080938 TI - Tonoplast-located GmCLC1 and GmNHX1 from soybean enhance NaCl tolerance in transgenic bright yellow (BY)-2 cells. AB - Genes encoding ion transporters that regulate ion homeostasis in soybean have not been carefully investigated. Using degenerate primers, we cloned a putative chloride channel gene (GmCLC1) and a putative Na+/H+ antiporter gene (GmNHX1) from soybean. Confocal microscopic studies using yellow fluorescent fusion proteins revealed that GmCLC1 and GmNHX1 were both localized on tonoplast. The expressions of GmCLC1 and GmNHX1 were both induced by NaCl or dehydration stress imposed by polyethylene glycol (PEG). Using mitochondrial integrity and cell death as the damage indicators, a clear alleviation under NaCl stress (but not PEG stress) was observed in both GmCLC1 and GmNHX1 transgenic cells. Using fluorescent dye staining and quenching, respectively, a higher concentration of chloride ion (Cl-) or sodium ion (Na+) was observed in isolated vacuoles in the cells of GmCLC1 and of GmNHX1 transgenic lines. Our result suggested that these vacuolar-located ion transporters function to sequester ions from cytoplasm into vacuole to reduce its toxic effects. PMID- 17080939 TI - Leaf growth and turgor in growing cells of maize (Zea mays L.) respond to evaporative demand under moderate irrigation but not in water-saturated soil. AB - To test whether the inhibition of leaf expansion by high evaporative demand is a result of hydraulic processes, we have followed both leaf elongation rate (LER) and cell turgor in leaves of maize plants either normally watered or in water saturated soil in which hydraulic resistance at the soil-root interface was abolished. Cell turgor was measured in situ with a pressure probe in the elongating zone of the first and sixth leaves, and LERs of the same leaves were measured continuously with transducers or by following displacements of marks along the growing leaves. Both variables displayed spatial variations along the leaf and positively correlated within the elongating zone. Values peaked at mid distance of this zone, where the response of turgor to evaporative demand was further dissected. High evaporative demand decreased both LER and turgor for at least 5 h, with dose-effect linear relations. This was observed in five genotypes with appreciable differences in turgor maintenance among genotypes. In contrast, the depressing effects of evaporative demand on both turgor and LER disappeared when the soil was saturated, thereby opposing a negligible resistance to water flow at the soil-root interface. These results suggest that the response of LER to evaporative demand has a hydraulic origin, enhanced by the resistance to water flux at the soil-root interface. They also suggest that turgor is not completely maintained under high evaporative demand, and may therefore contribute to the reductions in LER observed in non-saturated soils. PMID- 17080940 TI - Stem CO2 release under illumination: corticular photosynthesis, photorespiration or inhibition of mitochondrial respiration? AB - In illuminated stems and branches, CO2 release is often reduced. Many light triggered processes are thought to contribute to this reduction, namely photorespiration, corticular photosynthesis or even an inhibition of mitochondrial respiration. In this study, we investigated these processes with the objective to discriminate their influence to the overall reduction of branch CO2 release in the light. CO2 gas-exchange measurements of young birch (Betula pendula Roth.) branches (< 1.5 cm) performed under photorespiratory (20% O2) and non-photorespiratory (< 2%) conditions revealed that photorespiration does not play a pre-dominant role in carbon exchange. This suppression of photorespiration was attributed to the high CO2 concentrations (C(i)) within the bark tissues (1544 +/- 227 and 618 +/- 43 micromol CO2 mol(-1) in the dark and in the light, respectively). Changes in xylem CO2 were not likely to explain the observed decrease in stem CO2 release as gas-exchange measurements before and after cutting of the branches did not effect CO2 efflux to the atmosphere. Combined fluorescence and gas-exchange measurements provided evidence that the light dependent reduction in CO2 release can pre-dominantly be attributed to corticular refixation, whereas an inhibition of mitochondrial respiration in the light is unlikely to occur. Corticular photosynthesis was able to refix up to 97% of the CO2 produced by branch respiration, although it rarely led to a positive net photosynthetic rate. PMID- 17080941 TI - Complex adjustments of photosynthetic potentials and internal diffusion conductance to current and previous light availabilities and leaf age in Mediterranean evergreen species Quercus ilex. AB - Mature non-senescent leaves of evergreen species become gradually shaded as new foliage develops and canopy expands, but the interactive effects of integrated light during leaf formation (Q(int)G), current light (Q(int)C) and leaf age on foliage photosynthetic competence are poorly understood. In Quercus ilex L., we measured the responses of leaf structural and physiological variables to Q(int)C and Q(int)G for four leaf age classes. Leaf aging resulted in increases in leaf dry mass per unit area (M(A)), and leaf dry to fresh mass ratio (D(F)) and decreases in N content per dry mass (N(M)). N content per area (N(A)) was independent of age, indicating that decreases in N(M) reflected dilution of leaf N because of accumulation of dry mass (NA = N(M) M(A)). M(A), D(F) and N(A) scaled positively with irradiance, whereas these age-specific correlations were stronger with leaf growth light than with current leaf light. Area-based maximum ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) carboxylase activity (V(cmax)A), capacity for photosynthetic electron transport (J(max)A) and the rate of non-photorespiratory respiration in light (R(d)A) were also positively associated with irradiance. Differently from leaf structural characteristics, for all data pooled, these relationships were stronger with current light with little differences among leaves of different age. Acclimation to current leaf light environment was achieved by light-dependent partitioning of N in rate-limiting proteins. Mass-based physiological activities decreased with increasing leaf age, reflecting dilution of leaf N and a larger fraction of non-photosynthetic N in older leaves. This resulted in age-dependent modification of leaf photosynthetic potentials versus N relationships. Internal diffusion conductance (g(m)) per unit area (g(m)A) increased curvilinearly with increasing irradiance for two youngest leaf age classes and was independent of light for older leaves. In contrast, g(m) per dry mass (g(m)M) was negatively associated with light in current-year leaves. Greater photosynthetic potentials and moderate changes in diffusion conductance resulted in greater internal diffusion limitations of photosynthesis in higher light. Both area- and mass-based g(m) decreased with increasing leaf age. The decrease in diffusion conductance was larger than changes in photosynthetic potentials, leading to larger CO2 drawdown from leaf internal air space to chloroplasts (delta(c)) in older leaves. The increases in diffusion limitations in older leaves and at higher light scaled with age- and light-dependent increases in MA and D(F). Overall, our study demonstrates a large potential of foliage photosynthetic acclimation to changes in leaf light environment, but also highlights enhanced structural diffusion limitations in older leaves that result from leaf structural acclimation to previous rather than to current light environment and accumulation of structural compounds with leaf age. PMID- 17080942 TI - High expression in leaves of the zinc hyperaccumulator Arabidopsis halleri of AhMHX, a homolog of an Arabidopsis thaliana vacuolar metal/proton exchanger. AB - Zn hyperaccumulator plants sequester Zn into their shoot vacuoles. To date, the only transporters implicated in Zn sequestration into the vacuoles of hyperaccumulator plants are cation diffusion facilitators (CDFs). We investigated the expression in Arabidopsis halleri of a homolog of AtMHX, an A. thaliana tonoplast transporter that exchanges protons with Mg, Zn and Fe ions. A. halleri has a single copy of a homologous gene, encoding a protein that shares 98% sequence identity with AtMHX. Western blot analysis with vacuolar-enriched membrane fractions suggests localization of AhMHX in the tonoplast. The levels of MHX proteins are much higher in leaves of A. halleri than in leaves of the non accumulator plant A. thaliana. At the same time, the levels of MHX transcripts are similar in leaves of the two species. This suggests that the difference in MHX levels is regulated at the post-transcriptional level. In vitro translation studies indicated that the difference between AhMHX and AtMHX expression is not likely to result from the variations in the sequence of their 5' untranslated regions (5'UTRs). The high expression of AhMHX in A. halleri leaves is constitutive and not significantly affected by the metal status of the plants. In both species, MHX transcript levels are higher in leaves than in roots, but the difference is higher in A. halleri. Metal sequestration into root vacuoles was suggested to inhibit hyperaccumulation in the shoot. Our data implicate AhMHX as a candidate gene in metal accumulation or tolerance in A. halleri. PMID- 17080943 TI - O3 impacts on plant development: a meta-analysis of root/shoot allocation and growth. AB - The mechanism of O3 action on plants remains poorly characterized. Symptoms include visible lesions on the leaf surface, reduced growth and a hypothesized reduction in allocation of carbohydrate to roots. The generality of this latter phenomenon has not been demonstrated. Here, a meta-analysis is performed of all available experimental data, to test the hypotheses that O3 exposure of the shoot inhibits biomass allocation below ground (the root/shoot allometric coefficient, k) and inhibits whole-plant growth rate [relative growth rate (RGR)]. Both k and RGR were significantly reduced by O3 (5.6 and 8.2%, respectively). Variability in k was greater than in RGR, and both exhibited some positive as well as mostly negative responses. The effects on k were distinct from the effects on RGR. In some cases, k was reduced while RGR was unaffected. Slow-growing plants (small RGR) exhibited the largest declines in k. These observations may have mechanistic implications regarding O3 phytotoxicity. There were no effects of type of exposure chamber on sensitivity to O3. The analyses indicate that the O3 inhibition of allocation to roots is real and general, but variable. Further experiments are needed for under-represented plant groups, to characterize exceptions to this generalization and to evaluate O3--environment interactions. PMID- 17080944 TI - Expression of tobacco ethylene receptor NTHK1 alters plant responses to salt stress. AB - Ethylene has been regarded as a stress hormone involved in many stress responses. However, ethylene receptors have not been studied for the roles they played under salt stress condition. Previously, we characterized an ethylene receptor gene NTHK1 from tobacco, and found that NTHK1 is salt-inducible. Here, we report a further investigation towards the function of NTHK1 in response to salt stress by using a transgenic approach. We found that NTHK1 promotes leaf growth in the transgenic tobacco seedlings but affects salt sensitivity in these transgenic seedlings under salt stress condition. Differential Na+/K+ ratio was observed in the control Xanthi and NTHK1-transgenic plants after salt stress treatment. We further found that the NTHK1 transgene is also salt-inducible in the transgenic plants, and the higher NTHK1 expression results in early inductions of the ACC (1 aminocyclopropane-1-carboxylic acid) oxidase gene NtACO3 and ethylene responsive factor (ERF) genes NtERF1 and NtERF4 under salt stress. However, NTHK1 suppresses the salt-inducible expression of the ACC synthase gene NtACS1. These results indicate that NTHK1 regulates salt stress responses by affecting ion accumulation and related gene expressions, and hence have significance in elucidation of ethylene receptor functions during stress signal transduction. PMID- 17080945 TI - Evidence that differential gene expression between the halophyte, Thellungiella halophila, and Arabidopsis thaliana is responsible for higher levels of the compatible osmolyte proline and tight control of Na+ uptake in T. halophila. AB - Salt-sensitive glycophytes and salt-tolerant halophytes employ common mechanisms to cope with salinity, and it is hypothesized that differences in salt tolerance arise because of changes in the regulation of a basic set of salt tolerance genes. We explored the expression of genes involved in two key salt tolerance mechanisms in Arabidopsis thaliana and the halophytic A. thaliana relative model system (ARMS), Thellungiella halophila. Salt overly sensitive 1 (SOS1) is a plasma membrane Na+/H+ antiporter that retrieves and loads Na+ ions from and into the xylem. Shoot SOS1 transcript was more strongly induced by salt in T. halophila while root SOS1 was constitutively higher in unstressed T. halophila. This is consistent with a lower salt-induced rise in T. halophila xylem sap Na+ concentration than in A. thaliana. Thellungiella halophila contained higher unstressed levels of the compatible osmolyte proline than A. thaliana, while under salt stress, T. halophila accumulated more proline mainly in shoots. Expression of the A. thaliana ortholog of proline dehydrogenase (PDH), involved in proline catabolism, was undetectable in T. halophila shoots. The PDH enzyme activity was lower and T. halophila seedlings were hypersensitive to exogenous proline, indicating repression of proline catabolism in T. halophila. Our results suggest that differential gene expression between glycophytes and halophytes contributes to the salt tolerance of halophytes. PMID- 17080946 TI - Can fast-growing plantation trees escape biochemical down-regulation of photosynthesis when grown throughout their complete production cycle in the open air under elevated carbon dioxide? AB - Poplar trees sustain close to the predicted increase in leaf photosynthesis when grown under long-term elevated CO2 concentration ([CO2]). To investigate the mechanisms underlying this response, carbohydrate accumulation and protein expression were determined over four seasons of growth. No increase in the levels of soluble carbohydrates was observed in the young expanding or mature sun leaves of the three poplar genotypes during this period. However, substantial increases in starch levels were observed in the mature leaves of all three poplar genotypes grown in elevated [CO2]. Despite the very high starch levels, no changes in the expression of photosynthetic Calvin cycle proteins, or in the starch biosynthetic enzyme ADP-glucose pyrophosphorylase (AGPase), were observed. This suggested that no long-term photosynthetic acclimation to CO2 occurred in these plants. Our data indicate that poplar trees are able to 'escape' from long-term, acclimatory down regulation of photosynthesis through a high capacity for starch synthesis and carbon export. These findings show that these poplar genotypes are well suited to the elevated [CO2] conditions forecast for the middle of this century and may be particularly suited for planting for the long-term carbon sequestration into wood. PMID- 17080947 TI - An insect countermeasure impacts plant physiology: midrib vein cutting, defoliation and leaf photosynthesis. AB - One type of specialised herbivory receiving little study even though its importance has frequently been mentioned is vein cutting. We examined how injury to a leaf's midrib vein impairs gas exchange, whether impairment occurs downstream or upstream from injury, duration of impairment, compared the severity of midrib injury with non-midrib defoliation, and modelled how these two leaf injuries affect whole-leaf photosynthesis. Leaf gas exchange response to midrib injury was measured in five Asclepiadaceae (milkweed), one Apocynaceae (dogbane), one Polygonaceae and one Fabaceae species, which have been observed or reported to have midrib vein cutting injury in their habitats. Midrib vein injury impaired several leaf gas exchange parameters, but only downstream (distal) from the injury location. The degree of gas exchange impairment from midrib injury was usually more severe than from manually imposed and actual insect defoliation (non midrib), where partial recovery occurred after 28 d in one milkweed species. Non midrib tissue defoliation reduced whole-leaf photosynthetic activity mostly by removing photosynthetically active tissue, while midrib injury was most severe as the injury location came closer to the petiole. Midrib vein cutting has been suggested to have evolved as a countermeasure to deactivate induced leaf latex or cardenolide defences of milkweeds and dogbanes, yet vein cutting effects on leaf physiology seem more severe than the non-midrib defoliation the defences evolved to deter. PMID- 17080948 TI - The CBF1-dependent low temperature signalling pathway, regulon and increase in freeze tolerance are conserved in Populus spp. AB - The meristematic tissues of temperate woody perennials must acclimate to freezing temperatures to survive the winter and resume growth the following year. To determine whether the C-repeat binding factor (CBF) family of transcription factors contributing to this process in annual herbaceous species also functions in woody perennials, we investigated the changes in phenotype and transcript profile of transgenic Populus constitutively expressing CBF1 from Arabidopsis (AtCBF1). Ectopic expression of AtCBF1 was sufficient to significantly increase the freezing tolerance of non-acclimated leaves and stems relative to wild-type plants. cDNA microarray experiments identified genes up-regulated by ectopic AtCBF1 expression in Populus, demonstrated a strong conservation of the CBF regulon between Populus and Arabidopsis and identified differences between leaf and stem regulons. We studied the induction kinetics and tissue specificity of four CBF paralogues identified from the Populus balsamifera subsp. trichocarpa genome sequence (PtCBFs). All four PtCBFs are cold-inducible in leaves, but only PtCBF1 and PtCBF3 show significant induction in stems. Our results suggest that the central role played by the CBF family of transcriptional activators in cold acclimation of Arabidopsis has been maintained in Populus. However, the differential expression of the PtCBFs and differing clusters of CBF-responsive genes in annual (leaf) and perennial (stem) tissues suggest that the perennial driven evolution of winter dormancy may have given rise to specific roles for these 'master-switches' in the different annual and perennial tissues of woody species. PMID- 17080949 TI - Cell and leaf size plasticity in Arabidopsis: what is the role of endoreduplication? AB - Leaf area expansion is affected by environmental conditions because of differences in cell number and/or cell size. Increases in the DNA content (ploidy) of a cell by endoreduplication are related to its size. The aim of this work was to determine how cell ploidy interacts with the regulation of cell size and with leaf area expansion. The approach used was to grow Arabidopsis thaliana plants performing increased or decreased rounds of endoreduplication under shading and water deficit. The shading and water deficit treatments reduced final leaf area and cell number; however, cell area was increased and decreased, respectively. These differences in cell size were unrelated to alterations of the endocycle, which was reduced by these treatments. The genetic modification of the extent of endoreduplication altered leaf growth responses to shading and water deficit. An increase in the extent of endoreduplication in a leaf rendered it more sensitive to the shade treatment but less sensitive to water deficit conditions. The link between the control of whole organ and individual cell expansion under different environmental conditions was demonstrated by the correlation between the plasticity of cell size and the changes in the duration of leaf expansion. PMID- 17080950 TI - Exogenous supply of glutamine and active cytokinin to the roots reduces NO3- uptake rates in poplar. AB - The present study shows for the first time the influence of exogenously applied amino acids and cytokinin on the physiological and molecular aspects of N metabolism in poplar trees. In a short-term feeding experiment, glutamine or trans-zeatin riboside (tZR) was added directly to the nutrient solution. NO3- net uptake declined significantly in response to both treatments. Feeding with glutamine brought about an increase in concentrations of different amino compounds in the roots (glutamine, glutamate, alanine, gamma-amino butyric acid (GABA) and NH4+, which negatively correlated with the net NO3- uptake. The plants showed a reduction of cytosolic glutamine synthetase 1 (GS1) transcript level in the roots. In addition, glutamine feeding changed the root-to-shoot distribution on N assimilation in favour of the leaves and plant internal N cycling. tZR treatment resulted in expansion of zeatin-type (Z-type) cytokinins in the roots and increased nitrate reductase (NR)-mRNA level. The results indicate that both particular amino acids and active cytokinins are involved in the feedback regulation of N uptake and metabolism in poplar. We propose that inhibition of N uptake by cytokinins in poplar is more complex than that mediated by amino compounds, and other effectors are involved in this regulation. PMID- 17080951 TI - Thermal history regulates methylbutenol basal emission rate in Pinus ponderosa. AB - Methylbutenol (MBO) is a 5-carbon alcohol that is emitted by many pines in western North America, which may have important impacts on the tropospheric chemistry of this region. In this study, we document seasonal changes in basal MBO emission rates and test several models predicting these changes based on thermal history. These models represent extensions of the ISO G93 model that add a correction factor C(basal), allowing MBO basal emission rates to change as a function of thermal history. These models also allow the calculation of a new emission parameter E(standard30), which represents the inherent capacity of a plant to produce MBO, independent of current or past environmental conditions. Most single-component models exhibited large departures in early and late season, and predicted day-to-day changes in basal emission rate with temporal offsets of up to 3 d relative to measured basal emission rates. Adding a second variable describing thermal history at a longer time scale improved early and late season model performance while retaining the day-to-day performance of the parent single component model. Out of the models tested, the T(amb),T(max7) model exhibited the best combination of day-to-day and seasonal predictions of basal MBO emission rates. PMID- 17080952 TI - Spatial coordination of aluminium uptake, production of reactive oxygen species, callose production and wall rigidification in maize roots. AB - Aluminium (Al) toxicity associated with acid soils represents one of the biggest limitations to crop production worldwide. Although Al specifically inhibits the elongation of root cells, the exact mechanism by which this growth reduction occurs remains controversial. The aim of this study was to investigate the spatial and temporal dynamics of Al migration into roots of maize (Zea mays L.) and the production of the stress response compound callose. Using the Al-specific fluorescent probe morin, we demonstrate the gradual penetration of AI into roots. Al readily accumulates in the root's epidermal and outer cortical cell layers but does not readily penetrate into the inner cortex. After prolonged exposure times (12-24 h), Al had entered all areas of the root apex. The spatial and temporal accumulation of Al within the root is similarly matched by the production of the cell wall polymer callose, which is also highly localized to the epidermis and outer cortical region. Exposure to Al induced the rapid production of reactive oxygen species and induced a significant rigidification of the cell wall. Our results suggest that Al-induced root inhibition in maize occurs by rigidification of the epidermal layers. PMID- 17080953 TI - Involvement of gibberellins in the stem elongation of sun and shade ecotypes of Stellaria longipes that is induced by low light irradiance. AB - Plants from two ecotypes of Stellaria longipes, alpine (an open, sunny habitat) and prairie (where adjacent plants provide a shaded habitat), were grown under normal and reduced levels of photosynthetically active radiation (PAR). Growth under low PAR is significantly promoted in both ecotypes. When quantified by the stable isotope dilution method, endogenous gibberellins (GAs) (GA1, GA8, GA20, GA19) were significantly elevated under low PAR in both 'sun' and 'shade' ecotypes, as was GA53 in the shade ecotype. Changes in endogenous GA1 levels were significantly correlated with stem growth during a 28 d growth cycle and with relative growth rate (RGR) for height under low PAR for both ecotypes. Interestingly, under low irradiance PAR, changes (both increases and decreases) in GA8, the 2beta-hydroxylated 'inactive' catabolite of GA1, closely parallel bidaily stem growth changes for both ecotypes. Because the significantly greater stem elongation of both ecotypes in response to low irradiance PAR is associated with significant increases in the endogenous levels of five GAs (GA53, GA19, GA1, GA8) in the early 13-hydroxylation GA biosynthesis pathway (measured at days 7,14 and 21), we conclude that the low irradiance PAR has very likely induced an overall increase in GA biosynthesis. PMID- 17080954 TI - The effect of gibberellic acid on the response of leaf extension to low temperature. AB - The effect of cooling on leaf extension rate (LER) and on relative elemental growth rate (REGR) was measured in both gibberellic acid (GA)-responsive dwarf barley and in the same barley variety treated with GA. Seedlings were maintained at 20 degrees C while their leaf extension zone (LEZ) temperature was reduced either in steps to -6 degrees C in short-term cooling experiments, or to 10 degrees C for 48 h in long-term cooling experiments. Short-term cooling resulted in a biphasic response in LER, with a clear inflection point identified. Below this point, the activation energy for leaf extension becomes higher. The short term response of LER to cooling was altered by the application of GA, which resulted in a lower base temperature (Tb), inflection point temperature and activation energy for leaf extension. Both GA-treated and untreated seedlings were less sensitive to cooling maintained for a prolonged period, with LER making a partial recover over the initial 5 h. Although long-term cooling reduced maximum REGR, it resulted in a longer LEZ and an increase in the length of mature interstomatal cells in GA-treated and untreated seedlings. These changes in overall physiology appear to enhance the ability of the leaves to continue expansion at suboptimal temperatures. In both GA-treated and cold-acclimated tissue, the occurrence of a longer LEZ was associated with a lower temperature sensitivity in LER. PMID- 17080955 TI - Relationships between optically assessed polyphenols and chlorophyll contents, and leaf mass per area ratio in woody plants: a signature of the carbon-nitrogen balance within leaves? AB - Chlorophyll (Chl) and epidermal polyphenol (EPhen) contents were estimated in vivo using two optical leaf-clips, SPAD-502 and Dualex, respectively. The area based measurements were transformed into mass-based data by taking into account the leaf dry mass per area (LMA). Measurements were performed on forest trees and on saplings grown under controlled conditions. While LMA increased with irradiance along a vertical transect in a beech canopy or in saplings grown under different and increasing irradiance levels, mass-based EPhen (EPhen(m)) increased, whereas mass-based Chl (Chl(m)) decreased. This was a signature of a gradual switch of investment from protein into polyphenol production. A similar signature was obtained in saplings grown on nitrogen-deficient soil with respect to fertilized controls. However, nitrogen effects remained moderate compared to irradiance-induced effects. EPhen(m) and Chl(m) both declined with plant ageing induced increases in LMA, under all tested growth conditions. This was a signature of an accumulation of dry matter that diluted Chl and EPhen. The described competition between Chl and EPhen in leaves fits well with the predictions of the Protein Competition Model (PCM), that is, that the total leaf mass-based polyphenols content (Phen(t)) is controlled by the competition between protein and polyphenol biosynthetic pathways and its metabolic regulation. PMID- 17080956 TI - Coordinate up-regulation of carotenoid biosynthesis as a response to light stress in Synechococcus PCC7942. AB - Carotenoid biosynthesis is up-regulated by strong light in the cyanobacterium Synechococcus. By blocking off the pathway at the level of phytoene conversion, light-dependent accumulation of phytoene was observed. Real-time PCR studies demonstrated that four genes of the carotenogenic pathway are under transcriptional control. These were the genes encoding phytoene synthase, phytoene desaturase, zeta-carotene desaturase and beta-carotene hydroxylase. The transcript of the first three follow a similar kinetics, whereas the transcript of beta-carotene hydroxylase increased much faster upon transfer to high light. Promoter activities were determined with transcriptional fusions to chloramphenicol acyltransferase as reporter enzyme. The activity of the promoter of the phytoene desaturase/synthase operon was higher under strong light. PMID- 17080957 TI - Analysis of oxidative signalling induced by ozone in Arabidopsis thaliana. AB - We are using acute ozone as an elicitor of endogenous reactive oxygen species (ROS) to understand oxidative signalling in Arabidopsis. Temporal patterns of ROS following a 6 h exposure to 300 nL L(-1) of ozone in ozone-sensitive Wassilewskija (Ws-0) ecotype showed a biphasic ROS burst with a smaller peak at 4 h and a larger peak at 16 h. This was accompanied by a nitric oxide (NO) burst that peaked at 9 h. An analysis of antioxidant levels showed that both ascorbate (AsA) and glutathione (GSH) were at their lowest levels, when ROS levels were high in ozone-stressed plants. Whole genome expression profiling analysis at 1, 4, 8, 12 and 24 h after initiation of ozone treatment identified 371 differentially expressed genes. Early induction of proteolysis and hormone responsive genes indicated that an oxidative cell death pathway was triggered rapidly. Down-regulation of genes involved in carbon utilization, energy pathways and signalling suggested an inefficient defense response. Comparisons with other large-scale expression profiling studies indicated some overlap between genes induced by ethylene and ozone, and a significant overlap between genes repressed by ozone and methyl jasmonate treatment. Further, analysis of cis elements in the promoters of ozone-responsive genes also supports the view that phytohormones play a significant role in ozone-induced cell death. PMID- 17080958 TI - Functionality of resistance gene Hero, which controls plant root-infecting potato cyst nematodes, in leaves of tomato. AB - The expression of host genomes is modified locally by root endoparasitic nematode secretions to induce the development of complex cellular structures referred as feeding sites. In compatible interactions, the feeding sites provide the environment and nutrients for the completion of the nematode's life cycle, whereas in an incompatible (resistant) interaction, the host immune system triggers a plant cell death programme, often in the form of a hypersensitive reaction, which restricts nematode reproduction. These processes have been studied in great detail in organ tissues normally infected by these nematodes: the roots. Here we show that host leaves can support a similar set of programmed developmental events in the potato cyst nematode Globodera rostochiensis life cycle that are typical of the root-invading nematodes. We also show that a gene for-gene type specific disease resistance that is effective against potato cyst nematodes (PCN) in roots also operates in leaves: the expression of the resistance (R) gene Hero and members of its gene family in leaves correlates with the elicitation of a hypersensitive response only during the incompatible interaction. These findings, and the ability to isolate RNA from relevant parasitic stages of the nematode, may have significant implications for the identification of nematode factors involved in incompatible interactions. PMID- 17080959 TI - Agravitropic mutants of the moss Ceratodon purpureus do not complement mutants having a reversed gravitropic response. AB - New mutants of the moss Ceratodon purpureus have been isolated, which showed abnormal gravitropic responses. The apical cells of protonemal filaments of wild type strains respond to gravity by growing upwards and are well aligned to the gravity vector. This response only occurs in darkness. Mutants show a range of phenotypes. Some are insensitive to gravity, showing symmetrical growth, while others align to the gravity vector but orient growth downwards. A further class grows in darkness as though it were in light, showing insensitivity to gravity and continued chlorophyll synthesis. Somatic hybrids between mutants and wild type strains and between pairs of mutants have been selected using transgenic antibiotic resistance as selective markers. Hybrids between wild-type strains and all of the mutants have a wild-type phenotype, and so all mutants therefore have recessive phenotypes. Mutants comprise three complementation groups. One group has a single member, while another has three members. The third has at least 16 members and shows a complex pattern of complementation consistent with a single gene product functioning in both orientation and alignment to gravity, as well as contributing more than one subunit to the mature product. PMID- 17080960 TI - Oxygen dynamics during submergence in the halophytic stem succulent Halosarcia pergranulata. AB - This study elucidated O2 dynamics in shoots and roots of submerged Halosarcia pergranulata (Salicornioideae), a perennial halophytic stem succulent that grows on floodprone mudflats of salt lakes. Oxygen within shoots and roots was measured using microelectrodes, for plants when waterlogged or completely submerged, with shoots in light or in darkness, in a controlled environment. Net photosynthesis (PN) when underwater, at a range of dissolved CO2 concentrations, was measured by monitoring O2 production rates by excised stems. The bulky nature and apparently low volume of gas-filled spaces of the succulent stems resulted in relatively high radial resistance to gas diffusion. At ambient CO2, quasi-steady state rates of PN by excised succulent stems were estimated to be close to zero; nevertheless, in intact plants, underwater photosynthesis provided O2 to tissues and led to radial O2 loss (ROL) from the roots, at least during the first several hours (the time period measured) after submergence or when light periods followed darkness. The influence of light on tissue O2 dynamics was confirmed in an experiment on a submerged plant in a salt lake in south-western Australia. In the late afternoon, partial pressure of O2 (pO2) in the succulent stem was 23.2 kPa (i.e. approximately 10% above that in the air), while in the roots, it was 6.2 9.8 kPa. Upon sunset, the pO2 in the succulent stems declined within 1 h to below detection, but then showed some fluctuations with the pO2 increasing to at most 2.5 kPa during the night. At night, pO2 in the roots remained higher than in the succulent stems, especially for a root with the basal portion in the floodwater. At sunrise, the pO2 increased in the succulent stems within minutes. In the roots, changes in the pO2 lagged behind those in the succulent stems. In summary, photosynthesis in stems of submerged plants increased the pO2 in the shoots and roots so that tissues experience diurnal changes in the pO2, but O2 from the H2O column also entered submerged plants. PMID- 17080961 TI - Temperature dependence of nitrate reductase in the psychrophilic unicellular alga Koliella antarctica and the mesophilic alga Chlorella sorokiniana. AB - Temperature responses of nitrate reductase (NR) were studied in the psychrophilic unicellular alga, Koliella antarctica, and in the mesophilic species, Chlorella sorokiniana. Enzymes from both species were purified to near homogeneity by Blue Sepharose (Pharmacia, Uppsala, Sweden) affinity chromatography and high resolution anion-exchange chromatography (MonoQ; Pharmacia; Uppsala, Sweden). Both enzymes have a subunit molecular mass of 100 kDa, and K. antarctica NR has a native molecular mass of 367 kDa. NR from K. antarctica used both NADPH and NADH, whereas NR from C. sorokiniana used NADH only. Both NRs used reduced methyl viologen (MVH) or benzyl viologen (BVH). In crude extracts, maximal NADH and MVH dependent activities of cryophilic NR were found at 15 and 35 degrees C, respectively, and retained 77 and 62% of maximal activity, respectively, at 10 degrees C. Maximal NADH and MVH-dependent activities of mesophilic NR, however, were found at 25 and 45 degrees C, respectively, with only 33 and 23% of maximal activities being retained at 10 degrees C. In presence of 2 microM flavin adenine dinucleotide (FAD), activities of cryophilic NADH:NR and mesophilic NADH:NR were stable up to 25 and 35 degrees C, respectively. Arrhenius plots constructed with cryophilic and mesophilic MVH:NR rate constants, in both presence or absence of FAD, showed break points at 15 and 25 degrees C, respectively. Essentially, similar results were obtained for purified enzymes and for activities measured in crude extracts. Factors by which the rate increases by raising temperature 10 degrees C (Q10) and apparent activation energy (E(a)) values for NADH and MVH activities measured in enzyme preparations without added FAD differed slightly from those measured with FAD. Overall thermal features of the NADH and MVH activities of the cryophilic NR, including optimal temperatures, heat inactivation (with/without added FAD) and break-point temperature in Arrhenius plots, are all shifted by about 10 degrees C towards lower temperatures than those of the mesophilic enzyme. Transfer of electrons from NADH to nitrate occurs via all three redox centres within NR molecule, whereas transfer from MVH requires Mo-pterin prosthetic group only; therefore, our results strongly suggest that structural modification(s) for cold adaptation affect thermodynamic properties of each of the functional domains within NR holoenzyme in equal measure. PMID- 17080962 TI - Three grape CBF/DREB1 genes respond to low temperature, drought and abscisic acid. AB - The C-repeat (CRT)-binding factor/dehydration-responsive element (DRE) binding protein 1 (CBF/ DREB1) transcription factors control an important pathway for increased freezing and drought tolerance in plants. Three CBF/DREB1-like genes, CBF 1-3, were isolated from both freezing-tolerant wild grape (Vitis riparia) and freezing-sensitive cultivated grape (Vitis vinifera). The deduced proteins in V. riparia are 63-70% identical to each other and 96-98% identical to the corresponding proteins in V. vinifera. All Vitis CBF proteins are 42-51% identical to AtCBF1 and contain CBF-specific amino acid motifs, supporting their identification as CBF proteins. Grape CBF sequences are unique in that they contain 20-29 additional amino acids and three serine stretches. Agro infiltration experiments revealed that VrCBF1b localizes to the nucleus. VrCBF1a, VrCBF1b and VvCBF1 activated a green fluorescent protein (GFP) or glucuronidase (GUS) reporter gene behind CRT-containing promoters. Expression of the endogenous CBF genes was low at ambient temperature and enhanced upon low temperature (4 degrees C) treatment, first for CBF1, followed by CBF2, and about 2 d later by CBF3. No obvious significant difference was observed between V. riparia and V. vinifera genes. The expression levels of all three CBF genes were higher in young tissues than in older tissues. CBF1, 2 and 3 transcripts also accumulated in response to drought and exogenous abscisic acid (ABA) treatment, indicating that grape contains unique CBF genes. PMID- 17080963 TI - Synthesis of low molecular weight thiols in response to Cd exposure in Thlaspi caerulescens. AB - In this study, we investigated the accumulation of phytochelatins (PCs) and other low molecular weight (LMW) thiols in response to Cd exposure in two contrasting ecotypes differing in Cd accumulation. Using a root elongation test, we found that the highly accumulating ecotype Ganges was more tolerant to Cd than the low Cd-accumulation ecotype Prayon. L-buthionine-(S,R)-sulphoximine (BSO), a potent inhibitor of the gamma-glutamylcysteine synthetase gamma-ECS) (an enzyme involved in the PC biosynthetic pathway), increased the Cd sensitivity of Prayon, but had no effect on Ganges. Although PC accumulation increased in response to Cd exposure, no significant differences were observed between the two ecotypes. Cd exposure induced a dose-dependent accumulation of both Cys and a still unidentified LMW thiol in roots of both ecotypes. Root accumulation of Cys and this thiol was higher in Ganges than in Prayon; the ecotypic differences were more pronounced when the plants were treated with BSO. These findings suggest that PCs do not contribute to the Cd hypertolerance displayed by the Ganges ecotype of Thlaspi caerulescens, whereas Cys and other LMW thiols might be involved. PMID- 17080964 TI - Regulation of Arabidopsis thaliana 14-3-3 gene expression by gamma-aminobutyric acid. AB - The function in plants of the non-protein amino acid, gamma-aminobutyric acid (GABA) is poorly understood. In this study, we show that GABA down-regulates the expression of a large subset of 14-3-3 gene family members in Arabidopsis thaliana seedlings in a calcium, ethylene and abscisic acid (ABA)-dependent manner. Gene expression is not affected when seedlings are supplied with glutamate (GLU), a precursor of GABA. The repression of 14-3-3 gene expression by GABA is dependent on functional ethylene and ABA signalling pathways, because the response is lost in the etr1-1, abi1-1 and abi2-1 mutants. Calcium measurements show that in contrast to GLU, GABA does not elicit a cytoplasmic calcium elevation, suggesting that the GABA response is unlikely to be mediated by GLU receptors (GLRs), as has been suggested previously. We suggest that in addition to its role as a stress-related metabolite, GABA may regulate gene expression in A. thaliana, including members of the 14-3-3 gene family. PMID- 17080965 TI - Characterization of the Arabidopsis thermosensitive mutant atts02 reveals an important role for galactolipids in thermotolerance. AB - Plants are constantly challenged with various abiotic stresses in their natural environment. Elevated temperatures have a detrimental impact on overall plant growth and productivity. Many plants increase their tolerance to high temperatures through an adaptation response known as acquired thermotolerance. To identify the various mechanisms that plants have evolved to cope with high temperature stress, we have isolated a series of Arabidopsis mutants that are defective in the acquisition of thermotolerance after an exposure to 38 degrees C, a treatment that induces acquired thermotolerance in wild-type plants. One of these mutants, atts02, was not only defective in acquiring thermotolerance after the treatment, but also displayed a reduced level of basal thermotolerance in a 30 degrees C growth assay. The affected gene in atts02 was identified by positional cloning and encodes digalactosyldiacylglycerol synthase 1 (DGD1) (the atts02 mutant was, at that point, renamed dgd1-2). An additional dgd1 allele, dgd1-3, was identified in two other mutant lines displaying altered acquired thermotolerance, atts100 and atts104. Expression patterns of several heat shock proteins (HSPs) in heat-treated dgd1-2 homozygous plants were similar to those from identically treated wild-type plants, suggesting that the thermosensitivity in the dgd1-2 mutant was not caused by a defect in HSP induction. Lipid analysis of wild-type and mutant plants indicated a close correlation between the ability to acquire thermotolerance and the increases in digalactosyldiacylglycerol (DGDG) level and in the ratio of DGDG to monogalactosyldiacylglycerol (MGDG). Thermosensitivity in dgd1-2 and dgd1-3 was associated with (1) a decreased DGDG level and (2) an inability to increase the ratio of DGDG to MGDG upon exposure to a 38 degrees C sublethal temperature treatment. Our results suggest that the DGDG level and/or the ratio of DGDG to MGDG may play an important role in basal as well as acquired thermotolerance in Arabidopsis. PMID- 17080966 TI - The dehydrogenase-mediated recycling of NADPH is a key antioxidant system against salt-induced oxidative stress in olive plants. AB - NADPH is an important molecule in the redox balance of the cell. In this paper, using olive tissue cultures as a model of the function of the NADPH-generating dehydrogenases in the mechanism of oxidative stress induced by severe salinity conditions was studied. When olive (Olea europaea) plants were grown with 200 mM NaCl, a 40% reduction in leaf fresh weight was produced. The content of non enzymatic antioxidants such as ascorbate and glutathione was diminished between 20% to 39%, whereas the H2O2 content was increased threefold. In contrast, the analysis of the activity and protein contents of the main antioxidative enzymes showed a significant increase of catalase, superoxide dismutase and glutathione reductase. Overall, these changes strongly suggests that NaCl induces oxidative stress in olive plants. On the other hand, while the content of glucose-6 phosphate was increased almost eightfold in leaves of plants grown under salt stress, the content of NAD(P)H (reduced and oxided forms) did not show significant variations. Under salt stress conditions, the activity and protein contents of the main NADPH-recycling enzymes, glucose-6-phosphate dehydrogenase (G6PDH), isocitrate dehydrogenase (ICDH), malic enzyme (ME) and ferrodoxin-NADP reductase (FNR) showed an enhancement of 30-50%. In leaves of olive plants grown with 200 mM NaCl, analysis of G6PDH by immunocytochemistry and confocal laser scanning microscopy showed a general increase of this protein in epidermis, palisade and spongy mesophyll cells. These results indicate that in olive plants, salinity causes reactive oxygen species (ROS)-mediated oxidative stress, and plants respond to this situation by inducing different antioxidative enzymes, especially the NADPH-producing dehydrogenases in order to recycle NADPH necessary for the protection against oxidative damages. These NADP-dehydrogenases appear to be key antioxidative enzymes in olive plants under salt stress conditions. PMID- 17080967 TI - Of studies, syntheses, synopses, summaries, and systems: the "5S" evolution of information services for evidence-based health care decisions. PMID- 17080968 TI - Evaluating evidence-based practice performance. PMID- 17080969 TI - Aspirin plus dipyridamole was more effective than aspirin alone for preventing vascular events after minor cerebral ischemia. PMID- 17080970 TI - Clopidogrel plus aspirin was inferior to oral anticoagulation for preventing vascular events in atrial fibrillation. PMID- 17080971 TI - Early revascularization improved long-term survival after myocardial infarction with cardiogenic shock. PMID- 17080972 TI - Review: beta-blockers differ in their efficacy for preventing major cardiovascular events in younger and older patients. PMID- 17080973 TI - Review: early statin therapy does not reduce the composite endpoint of death, MI, or stroke in acute coronary syndromes. PMID- 17080974 TI - Atorvastatin did not prevent cardiovascular events in type 2 diabetes. PMID- 17080975 TI - N-acetylcysteine prevented contrast-medium-induced nephropathy in primary angioplasty. PMID- 17080976 TI - Review: a low-protein diet delays end-stage renal disease or death in stage 3 to 5 chronic kidney disease. PMID- 17080977 TI - Review: anti-tumor necrosis factor antibody therapy for rheumatoid arthritis increases risk for serious infection and malignancy. PMID- 17080978 TI - Review: selective COX-2 inhibitors increase vascular events more than placebo and naproxen, but not more than other NSAIDs. PMID- 17080979 TI - Adjunctive inhaled insulin before meals improved glycemic control more than adjunctive metformin in type 2 diabetes mellitus. PMID- 17080981 TI - A conservative fluid management strategy did not affect risk for death but shortened duration of ventilation in acute lung injury. PMID- 17080980 TI - Oral decontamination with chlorhexidine reduced ventilator-associated pneumonia in high-risk patients. PMID- 17080982 TI - Pulmonary artery catheters and central venous catheters did not differ for mortality in acute lung injury. PMID- 17080983 TI - Review: human parathyroid hormone reduces fractures and increases bone mineral density in severe osteoporosis. PMID- 17080984 TI - Raloxifene and tamoxifen had similar efficacy for preventing invasive breast cancer in women at increased risk. PMID- 17080985 TI - Raloxifene produced both harms and benefits in postmenopausal women, with no reduction in cardiovascular disease risk. PMID- 17080986 TI - Review: opioids are more effective than placebo but not other analgesics for chronic noncancer pain. PMID- 17080987 TI - Review: prophylactic interventions reduce oral mucositis in patients with cancer receiving chemotherapy or radiation therapy. PMID- 17080988 TI - Multidetector CTA with venography was more sensitive for diagnosing pulmonary embolism than CTA alone. PMID- 17080989 TI - Review: noninvasive imaging techniques may be useful for diagnosing 70% to 99% carotid stenosis in symptomatic patients. PMID- 17080990 TI - 1-year mortality after first hospitalization for heart failure was similar in patients with preserved or reduced ejection fraction. PMID- 17080991 TI - Review: healthy kidney donors may have a long-term increase in blood pressure beyond that associated with normal aging. PMID- 17080992 TI - Review: good adherence (compared with poor adherence) to drug therapy is associated with a reduction in mortality. PMID- 17080993 TI - A nurse-led clinical pathway reduced hospitalizations in nursing home residents with pneumonia. PMID- 17080994 TI - Sulfated meroterpenoids from the Brazilian sponge Callyspongia sp. are inhibitors of the antileishmaniasis target adenosine phosphoribosyl transferase. AB - Three new disulfated meroterpenoids, ilhabelanol (1), ilhabrene (2), and isoakaterpin (3), have been isolated from extracts of the Brazilian marine sponge Callyspongia sp. Isoakaterpin (3) inhibits Leishmania spp. adenosine phosphoribosyl transferase with an IC50 of 1.05 microM. The structures of 1, 2, and 3 were elucidated by analysis of one- and two-dimensional NMR data. Ilhabelanol (1) and ilhabrene (2) both have unprecedented meroterpenoid carbon skeletons. PMID- 17080995 TI - Synthesis of structurally diverse bis-peptide oligomers. AB - We have developed second-generation monomers 1 and 2 and improved conditions for rapidly and simultaneously closing multiple diketopiperazines on solid support. These new conditions involve either the microwave heating of a suspension of solid-supported amino-tetrafluoropropyl esters in acetic acid/triethylamine catalyst solution or continuous flow of catalyst solution through the resin, heated in a flow cell apparatus. We demonstrate that the new monomers 1 and 2 can be combined with the new conditions easily to synthesize previously inaccessible hetero and homo spiro ladder oligomers 3 and 4 and others. PMID- 17080996 TI - Supramolecular structures from lysine peptides and carbon dioxide. AB - The design, synthesis, and characterization of novel linear and cross-linked supramolecular polymers that are easily available from biologically friendly lysine peptides and carbon dioxide (CO2) are reported here. Polymeric structures 5, 6, and 19 readily form from peptides 2, 3, and 15, respectively, at ambient temperatures by simply bubbling CO2 through their solutions in apolar organic solvents (CHCl3, benzene) and even in the presence of 10% MeOH. The resulting gels can be easily isolated from solution, dried, and stored refrigerated for several months. At the same time, they may thermally release CO2 and convert back to the corresponding monomers. As a consequence, their structures and physical properties are switchable. They may also trap, store, and release foreign molecules. The typical entrapment procedure was demonstrated for tripeptide 3, CO2, and the commercially available dye coumarin 2. PMID- 17080997 TI - Hangman salen platforms containing two xanthene scaffolds. AB - A synthetic strategy for the construction of chiral salen ligands bearing two rigid xanthene spacers functionalized with carboxylic acid and ester groups is presented. Suzuki cross-coupling methodology is used to furnish the appropriately functionalized xanthene spacers to a salicylaldehyde, which is subsequently condensed with (1R,2R)-(-)-1,2-diaminocyclohexane to produce salen ligands featuring an expandable molecular cleft capable of multiple hydrogen-bonding interactions in addition to metallosalen oxidation chemistry. The ability of these "Hangman" platforms to support multielectron chemistry mediated by proton coupled electron transfer (PCET) is established by their proclivity to promote the catalytic disproportionation of hydrogen peroxide to oxygen and water via a high-valent metal oxo. Within this functionalized Hangman framework, the stereochemistry of the cyclohexyl backbone of the salen platform is revealed in the epoxidation of 1,2-dihydronaphthalene by the metal oxo. PMID- 17080998 TI - Condensation of alpha-aroylketene dithioacetals and 2-hydroxyarylaldehydes results in facile synthesis of a combinatorial library of 3-aroylcoumarins. AB - A facile, convenient, efficient, and high yielding synthesis of a combinatorial library of 3-aroylcoumarins has been developed by the condensation of easily available alpha-aroylketene dithioacetals (AKDTAs) and 2-hydroxybenzaldehydes (salicylaldehydes)/2-hydroxy-1-naphthaldehyde in the presence of catalytic amount of piperidine in THF reflux. The condensation of ferrocene derived alpha aroylketene dithioacetal and 2-hydroxybenzaldehyde furnished coumarin installed on a ferrocene platform. PMID- 17080999 TI - Biosynthesis of 13-desmethyl spirolide C by the dinoflagellate Alexandrium ostenfeldii. AB - Biosynthetic origins of the cyclic imine toxin 13-desmethyl spirolide C were determined by supplementing cultures of the toxigenic dinoflagellate Alexandrium ostenfeldii with stable isotope-labeled precursors [1,2-13C2]acetate, [1 13C]acetate, [2-13CD3]acetate, and [1,2-13C2,15N]glycine and measuring the incorporation patterns by 13C NMR spectroscopy. Despite partial scrambling of the acetate labels, the results show that most carbons of the macrocycle are polyketide-derived and that glycine is incorporated as an intact unit into the cyclic imine moiety. This work represents the first conclusive evidence that such cyclic imine toxins are polyketides and provides support for biosynthetic pathways previously defined for other polyether dinoflagellate toxins. PMID- 17081000 TI - Emission from charge recombination during the pulse radiolysis of 9-cyano-10 phenylethynylanthracenes with donor and acceptor substituents. AB - Emission from 9-cyano-10-phenylanthracene and 9-cyano-10-phenylethynylanthracenes having donor and acceptor substituents (RA = PA, PEA, OEA, NEA, and DEA) was studied with the time-resolved fluorescence measurement during the pulse radiolysis of RAs in benzene (Bz). PA and DEA showed only monomer emission, while other RAs (PEA, OEA, and NEA) showed both monomer and excimer emissions with much lower intensities. On the basis of the steady-state and transient absorption and emission measurements, the formation of RA in the singlet excited state ((1)RA*) can be attributed to the charge recombination between RA radical cation and anion (RA*+ and RA*-, respectively) which are initially generated from the radiolytic reaction in Bz. It is expected that for PA with a twisted geometry, the charge recombination between PA*+ and PA*- occurs to give (1)PA* during the pulse radiolysis in Bz. For PEA and OEA, pi-stacking interaction is possible for the formation of an encounter complex during the charge recombination between RA*+ and RA*-. For NEA, it is expected that NEA*+ and NEA*- collide neck-to-neck to generate the excimer due to the twisted geometry. For DEA, a considerably twisted structure is assumed to give (1)DEA* with strong ICT character but not (1)(DEA)2* because of the bulky donor substituent. PMID- 17081001 TI - Homogeneous decatungstate-catalyzed photooxygenation of tetrasubstituted alkenes: a deuterium kinetic isotope effect study. AB - The decatungstate W10O32(4-) homogeneous photocatalyzed oxygenation of tetrasubstituted alkenes has been mechanistically studied. In all cases, allylic hydroperoxides are the major products. The primary inter- and intramolecular as well as the remote delta-secondary deuterium kinetic isotope effects for the photooxidation of the 2,3-dimethyl-2-butene and 1,1,1-trideuterio-7-methyl-2 (trideuteriomethyl)octa-2,6-diene along with product analysis suggest a hydrogen abstraction in the rate-determining step. For comparison, singlet oxygen photosensitized oxidations of the above substrates were also studied. PMID- 17081002 TI - Divergent chemical synthesis of prolines bearing fluorinated one-carbon units at the 4-position via nucleophilic 5-endo-trig cyclizations. AB - N-[3-(Trifluoromethyl)homoallyl]sulfonamides, prepared via ring opening of (S) glycidyl ethers or 2-aryloxiranes with 1-(trifluoromethyl)vinyllithium, underwent intramolecular addition or S(N)2'-type reaction in the normally disfavored 5-endo trig fashion, leading to 2-substituted 4-(trifluoromethyl)- or 4 (difluoromethylene)pyrrolidines. Both alpha- and beta-face-selective hydrogenation of the 4-difluoromethylene group afforded syn- and anti-4 (difluoromethyl)pyrrolidines, respectively. These sequences, followed by the oxidation of a 2-hydroxymethyl or 2-aryl group, successfully provided prolines with a trifluoromethyl, difluoromethylene, or difluoromethyl group at the 4 position, including optically active prolines. PMID- 17081003 TI - Reaction of alpha-(N-carbamoyl)alkylcuprates with enantioenriched propargyl electrophiles: synthesis of enantioenriched 3-pyrrolines. AB - Enantioenriched propargyl mesylates or perfluorobenzoates react with alpha-(N carbamoyl)alkylcuprates to afford scalemic alpha-(N-carbamoyl) allenes which undergo N-Boc deprotection and AgNO3-promoted cyclization to afford N-alkyl-3 pyrrolines. The synthetic sequence proceeds under optimal conditions with no loss of enantiopurity relative to the starting propargyl alcohols prepared by asymmetric addition of terminal alkynes to aldehydes. PMID- 17081004 TI - Total synthesis of (-)-normalindine via addition of metalated 4-methyl-3 cyanopyridine to an enantiopure sulfinimine. AB - A concise total asymmetric synthesis of the tetrahydronaphthyridine alkaloid (-) normalindine has been accomplished via the addition of a laterally metalated 4 methyl-3-cyanopyridine to a sulfinimine (N-sulfinyl imine) as the key step. PMID- 17081005 TI - Divergent synthetic strategy leading to structurally diverse pyrrolidines and piperidines from common gamma-aminoalkyl butenolide and aldehyde precursors. AB - Condensation between aldehydes and the secondary amino function of 5 (aminoalkyl)furan-2(5H)-ones, obtained by the silyloxyfuran dienolate addition to imine-type derivatives, produces either aminoalkylbenzotriazoles or 1,2,3,4 tetrahydropyridines. The former can be reduced with SmI2 to generate alpha aminoalkyl radicals that are trapped by the alpha,beta-unsaturated lactone moiety yielding substituted pyrrolidines diastereoselectively, while catalytic hydrogenation of the latter affords isomeric piperidine analogues. Alternatively, SmI2-promoted reduction of tetrahydropyridines in the presence of acid also leads to intermediate alpha-aminoalkyl radicals that participate in inter- or intramolecular olefin addition reactions. Further manipulation of the lactone functionality in various ways gives access to a number of interesting derivatives based upon either a pyrrolidine or a piperidine structural motif. As a result, a high degree of structural diversity is obtained in a few steps starting from a common set of simple materials. PMID- 17081006 TI - ESI-MS detection of proposed reaction intermediates in the air-promoted and ligand-modulated oxidative Heck reaction. AB - Electrospray ionization mass spectrometry (ESI-MS) and subsequent MS/MS analyses were used to directly detect palladium-containing cationic reaction intermediates in a ligand controlled palladium(II)-catalyzed oxidative Heck arylation. All potential intermediates were observed as dmphen-ligated palladium(II) species, suggesting that the dmphen bidentate ligand is attached to the metal center during the entire catalytic cycle. The study supports previous mechanistic propositions and provides new information regarding the composition of aryl containing Pd(II) complexes in an ongoing oxidative Heck reaction. In addition, sodium acetate was found to be a useful base alternative to previously used tertiary amines. PMID- 17081007 TI - Synthesis of highly functionalized macrocycles by the peripheral functionalization of macrocyclic diimines. AB - The easily available macrocyclic diimines 4-7 can be stereoselectively transformed to macrocyclic bis-beta-amino acids 13-17, macrocyclic bisazetidines 18-20, and macrocyclic bisamides 21 and 22 by means of the corresponding bis-beta lactam scaffolds 8-12. These key intermediates are available through standard Staudinger reaction and obtained as the cis-cis diastereomers, exclusively. An interesting relation between the proximity of the reactive C=N bonds and the selectivity in the formation of the bis-beta-lactams 8-12 is observed. Thus, diimine 4 leads to low selectivities, producing a 1:1 mixture of cis-syn-cis and cis-anti-cis diastereomers, while diimines 5-7 having the diimine sites more separated lead almost exclusively to the cis-anti-cis diastereomers. The stereochemistry of all the products was unambiguously assigned by X-ray diffraction analysis of compounds cis-syn-cis 8 and cis-anti-cis 12-Co2CO6 complex. PMID- 17081008 TI - Solid-state nitrogen-15 NMR and quantum chemical study of N,N-dimethylaniline derivatives. AB - In this study the components of the nitrogen chemical shift (CS) tensor are examined for a series of para substituted N,N-dimethylaniline derivatives. This is done through measurement of the 15N NMR spectra of powder samples and through quantum chemical calculations on the isolated molecules. Experiments and calculations show that the isotropic CS, delta(iso), decreases with increasing electron donating ability of the para substituent, in agreement with previous solution studies. More importantly, this study shows that this decrease in the isotropic (solution) CS is due to decreasing values of the CS tensor component delta(11) and component delta(33). The component delta(22) is essentially invariant to the electron donating/withdrawing ability of the para substituent. Through Ramsey's theory of nuclear magnetic shielding, it can be seen that the variation in delta(11) and delta(33), and hence delta(iso), is due to changes in the n-pi* and the sigma-pi* energy gaps in N,N-dimethylaniline. This, in turn, is a result of the change in the energy of the pi* molecular orbital with change in the pi-electron donating ability of the para substituent. The effects of nitrogen inversion on the components of the nitrogen CS tensor components are also discussed. This study also shows the feasibility of performing 15N cross polarization experiments on nonspinning powder samples at natural isotopic abundance. PMID- 17081009 TI - Tunable capsule space: self-assembly of hemispherical cavitands with hydrogen bonding linkers. AB - Fine and/or drastic tuning of capsule space has been attained by alteration of the hydrogen-bonding linker and/or hemispherical cavitand, respectively. Two molecules of tetracarboxyl-cavitand 1 or tetrakis(4-carboxyphenyl)-cavitand 2 as a hemisphere and four molecules of 2-aminopyrimidine (2-AP) or tetrahydro-2 pyrimidinone (THP) as an equatorial hydrogen-bonding linker self-assemble into a capsule [(1)2.(2-AP)4] (3), [(1)2.(THP)4] (4), [(2)2.(2-AP)4] (5), or [(2)2.(THP)4] (6), respectively, via 16 hydrogen bonds. These capsules provide isolated nanospace and can encapsulate one guest molecule (7-13) in solution. Each capsule has a different cavity size and shows particular guest selectivity on the competitive encapsulation experiments. PMID- 17081010 TI - Masked imidazolyl-dipyrromethanes in the synthesis of imidazole-substituted porphyrins. AB - Imidazole-substituted metalloporphyrins are valuable for studies of self-assembly and for applications where water solubility is required. Rational syntheses of porphyrins bearing one or two imidazol-2-yl or imidazol-4-yl groups at the meso positions have been developed. The syntheses employ dipyrromethanes, 1 acyldipyrromethanes, and 1,9-diacyldipyrromethanes bearing an imidazole group at the 5-position. The polar, reactive imidazole unit was successfully masked by use of (1) the 2-(trimethylsilyl)ethoxymethyl (SEM) group at the imidazole pyrrolic nitrogen, and (2) a dialkylboron motif bound to the pyrrole of the dipyrromethane and coordinated to the imidazole imino nitrogen. The nonpolar nature of such doubly masked imidazolyl-dipyrromethanes facilitated handling. Selected masked dipyrromethanes were characterized by 11B and 15N NMR spectroscopy. Five distinct methods were examined to obtain trans-A2B2-, trans-AB2C-, and trans-AB porphyrins. Each porphyrin contained one or two SEM-protected imidazole units. The SEM group could be removed with TBAF or HCl. Two zinc(II) porphyrins and a palladium(II) porphyrin bearing a single imidazole moiety were prepared and subjected to alkylation (with ethyl iodide, 1,3-propane sultone, or 1,4-butane sultone) to give water-soluble imidazolium- porphyrins. This work establishes the foundation for the rational synthesis of a variety of porphyrins containing imidazole units. PMID- 17081011 TI - General route to racemic and enantiomeric carbo- and heterocyclic vinyl sulfoxides via tandem Michael addition/Horner olefination of alpha phosphorylvinyl sulfoxides. AB - A new one-pot synthesis of heterocyclic and carbocyclic vinyl sulfoxides has been developed which involves reaction of alpha-phosphorylvinyl sulfoxides with carbonyl compounds bearing oxygen, nitrogen, and carbon nucleophilic centers. Use of optically active alpha-phosphorylvinyl p-tolyl sulfoxides in this tandem Michael addition/Horner olefination reaction leads to the corresponding optically active cyclic sulfoxides. In this way, a variety of optically active chromene, pyrrolizine, chinoline, and cyclopentene sulfoxides have been efficiently prepared. PMID- 17081012 TI - Nonmevalonate terpene biosynthesis enzymes as antiinfective drug targets: substrate synthesis and high-throughput screening methods. AB - The nonmevalonate isoprenoid pathway is an established target for antiinfective drug development. This paper describes high-throughput methods for the screening of 2C-methyl-D-erythritol synthase (IspC protein), 4-diphosphocytidyl-2C-methyl-D erythritol synthase (IspD protein), 4-diphosphocytidyl-2C-methyl-D-erythritol kinase (IspE protein), and 2C-methyl-D-erythritol 2,4-cyclodiphosphate synthase (IspF protein) against large compound libraries. The assays use up to three auxiliary enzymes. They are all monitored photometrically at 340 nm and are robust as documented by Z-factors of >or=0.86. 13C NMR assays designed for hit verification via direct detection of the primary reaction product are also described. Enzyme-assisted methods for the preparation, on a multigram scale, of isoprenoid biosynthesis intermediates required as substrates for these assays are reported. Notably, these methods enable the introduction of single or multiple 13C labels as required for NMR-monitored assays. The preparation of 4 diphosphosphocytidyl-2C-methyl-D-erythritol 2-phosphate in multigram quantities is described for the first time. PMID- 17081013 TI - Effect of 2,6-disubstituted aryl groups on acyclic conformation: preference for an antiperiplanar orientation of the geminal and vicinal hydrogens. AB - Multiple X-ray crystallographic and 1H NMR spectroscopic studies demonstrate that 2,6-disubstituted aryl groups exert a strong effect on acyclic conformation, inducing the geminal and vicinal hydrogens on the adjacent sp3-sp3 C-C bond to adopt an antiperiplanar orientation. Twenty-one examples comprising nitrile, ketone, and dithiane aldols are provided. A search of the Cambridge Structural Database uncovered an additional 11 examples of this effect. This preference causes some unanticipated remote effects on acyclic conformations: in anti nitrile aldols, this effect causes 2,6-disubstituted aryls to generally prefer a gauche, rather than antiperiplanar, relationship to the largest vicinal groups. X ray crystallography, 1H-1H NOESY spectroscopy, and computation demonstrate that minimization of allylic 1,3-strain and syn-pentane-like interactions work together in establishing this conformational preference. PMID- 17081014 TI - Simple synthesis of some pentafluoropropenyl derivatives of pyrimidine and purine based on addition-elimination reaction. AB - Various pentafluoropropenyl derivatives of pyrimidine and purine bases have been obtained in good to high yield. The procedure involves the reaction of appropriate lithium derivatives prepared from both electron-rich and electron poor pyrimidines, with the hexafluoropropene at a low temperature, via an addition-elimination process. Organolithiums of pyrimidine and purine bases give addition-elimination products as E/Z mixtures, whereas the products of the reaction of lithium amide of protected inosine with hexafluoropropene contain traces of an addition product as well as the stable perfluoroenamine. The methodology proposed allows a series of perfluorovinyl nucleobases to be obtained quickly and conveniently. PMID- 17081015 TI - Using Kamlet-Taft solvent descriptors to explain the reactivity of anionic nucleophiles in ionic liquids. AB - In this paper, we report the effect of ionic liquids on substitution reactions using a variety of anionic nucleophiles. We have combined new studies of the reactivity of polyatomic anions, acetate, trifuoroacetate, cyanide, and thiocyanide, with our previous studies of the halides in [C4C1py][Tf2N], [C4C1py][TfO], and [C4C1im][Tf2N] (where [C4C1im]+ is 1-butyl-3-methylimidazolium and [C4C1py]+ is 1-butyl-1-methylpyrrolidinium) and compared their reactivities, k2, to the same reactions in the molecular solvents dichloromethane, dimethylsulfoxide, and methanol. The Kamlet-Taft solvent descriptors (alpha, beta, pi) have been used to analyze the rates of the reactions, which were found to have a strong inverse dependency on the alpha value of the solvent. This result is attributed to the ability of the solvent to hydrogen bond to the nucleophile, so reducing its reactivity. The Eyring activation parameters (DeltaH++ and DeltaS++), while confirming the reaction mechanism, do not offer obvious correlations with the Kamlet-Taft solvent descriptors. PMID- 17081016 TI - Diastereoselective synthesis of enantiopure morpholines by electrophilic selenium induced 6-exo cyclizations on chiral 3-allyl-2-hydroxymethylperhydro-1,3 benzoxazine derivatives. AB - Enantiopure morpholine derivatives have been prepared by selenocyclofunctionalization of chiral 3-allyl-2-hydroxymethyl-substituted perhydro-1,3-benzoxazine derivatives. The cyclization occurs in high yields and diastereoselection, although the temperature of the reaction and the structure of the substituent at C-2 and the substitution pattern of the double bond can modify the regio- and stereochemistry of the final products. PMID- 17081017 TI - Epimerization reaction of a substituted vinylcyclopropane catalyzed by ruthenium carbenes: mechanistic analysis. AB - A novel ruthenium carbene-catalyzed epimerization of vinylcyclopropanes is reported. The reaction rate strongly depends on the presence of ruthenium ligands in solution. When the first-generation Grubbs catalyst is employed, a 5.3:1 equilibrium ratio of epimers is established quickly, but when a first-generation Hoveyda catalyst is employed, epimerization is observed only if an additional phosphine or nitrogen ligand is added. NMR and kinetic studies suggest that the isomerization reaction occurs through the intermediacy of a ruthenacyclopentene. The observation suggests that cyclopropylmethylidene ruthenium carbenes of synthetic utility may be accessible via ruthenacyclopentenes obtained via other routes. PMID- 17081018 TI - Addition of difluorocarbene to 4',5'-unsaturated nucleosides: synthesis and deoxygenation reactions of difluorospirocyclopropane nucleosides. AB - Synthetic routes to 4'-(2,2-difluorospirocyclopropane) analogues of adenosine, cytidine, and uridine are described. Treatment of 2',3'-O-isopropylidene-4',5' unsaturated compounds derived from adenosine and uridine with difluorocarbene (generated from PhHgCF3 and NaI) gave diastereomeric mixtures of the 2,2 difluorospirocyclopropane adducts. Stereoselectivity resulting from hindrance by the isopropylidene group favored addition at the beta face. Removal of base and sugar protecting groups gave new difluorospirocyclopropane nucleoside analogues. The protected uridine analogue was converted into its cytidine counterpart via a 4-(1,2,4-triazol-1-yl) intermediate. Stannyl radical-mediated deoxygenation of the 3'-O-TBS-2'-thionocarbamate derivatives gave the 2'-deoxy products of direct hydrogen transfer. In contrast, identical treatment of the 2'-O-TBS-3' thionocarbamate isomers resulted in opening of the vicinal difluorocyclopropane ring upon generation of a C3' radical followed by homoallylic hydrogen transfer to give 4'-(1,1-difluoroethyl)-3',4'-unsaturated nucleoside derivatives. Structural aspects and biological effect considerations are discussed. PMID- 17081019 TI - An improved total synthesis of (+)-macroline and alstonerine as well as the formal total synthesis of (-)-talcarpine and (-)-anhydromacrosalhine-methine. AB - An intramolecular Pd-catalyzed alpha-vinylation process is described. This cyclization has been employed for the enantiospecific total synthesis of gram quantities of both (+)-macroline 3 and the macroline equivalent 4. This sequence is compared to the enolate-driven cross-coupling process. The intermediate 4 was also converted into (-)-alstonerine 1 via modification of an intramolecular Tsuji Wacker oxidation. This sequence resulted in an improved total synthesis of (-) talcarpine 5 and (-)-anhydromacrosalhine-methine 6 as well. PMID- 17081020 TI - Synthetic studies on perophoramidine and the communesins: construction of the vicinal quaternary stereocenters. AB - An efficient synthetic strategy for installation of the two vicinal quaternary carbon centers of the communesins is reported. Key steps include the O allylation/Claisen rearrangement of spirolactone systems, which are formed by tandem intramolecular Heck cyclization/carbonylation. Substituent and solvent effects on the stereochemical outcome of the Claisen rearrangements have been examined. The stereochemical assignment of the allyl spirolactone previously reported as 17 has now been revised to 31, which has the communesin relative configuration at the quaternary carbons. Key C-allyl spirolactone 59 bearing functional handles required for the communesin core has been constructed with a 9.8:1 diastereomer ratio. PMID- 17081021 TI - Regiospecific N9 alkylation of 6-(heteroaryl)purines: shielding of N7 by a proximal heteroaryl C-H1. AB - Purine alkylations have been plagued with formation of mixtures of N9 (usually desired), N7, and other regioisomers. We have developed methods for synthesis of 6-(azolyl)purine derivatives whose X-ray crystal structures show essentially coplanar conformations of the linked azole-purine rings. Such ring orientations position the C-H of the azole above N7 of the purine, which results in protection of N7 from alkylating agents. Treatment of 6-(2-butylimidazol-1-yl)-2 chloropurine (9) with sodium hydride in DMF followed by addition of ethyl iodide resulted in exclusive formation of 6-(2-butylimidazol-1-yl)-2-chloro-9 ethylpurine (10), whereas identical treatment of 2-chloro-6-(4,5-diphenylimidazol 1-yl)purine (11) produced a regioisomeric mixture 12/13 (N9/N7, approximately 5:1). The linked imidazole and purine rings are coplanar in 9 (the butyl side chain is extended away from the purine ring and C-H is over N7) but are rotated approximately 57 degrees in 11, and the more bulky azole substituent in 11 did not prevent formation of the minor N7 regioisomer 13. Access to various regioisomerically pure 9-alkylpurines is now readily available. PMID- 17081022 TI - On the mechanism of the conversion of methanol to 2,2,3-trimethylbutane (triptane) over zinc iodide. AB - Methanol is converted to a mixture of hydrocarbons by reaction with zinc iodide at 200 degrees C with one highly branched alkane, 2,2,3-trimethylbutane (triptane), being obtained in surprisingly high selectivity. Mechanistic studies implicate a two-stage process, the first involving heterogeneously catalyzed formation of a carbon-carbon-bonded species, probably ethylene, that undergoes homogeneously catalyzed sequential cationic methylation to higher hydrocarbons. The first stage can be bypassed by addition of olefins, higher alcohols, or arenes, which act as initiators. Rationales for the particular activity of zinc iodide and for the selectivity to triptane are proposed. PMID- 17081023 TI - Experimental and theoretical studies on radical trifluoromethylation of titanium ate and lithium enolates. AB - The radical trifluoromethylation of Ti ate and Li enolates has been investigated by both experiments and density functional (UB3LYP/6-311+G//UB3LYP/6-31+G*) calculations. Radical CF3 addition to the enolates proceeds in a highly exothermic manner without significant reaction barriers in both Ti ate and Li enolates. There are two possible reaction paths after the addition of CF3 radical in the case of Ti ate enolate; one is the elimination of Ti(III) from the ketyl radical intermediate and the other is the direct reaction of the ketyl radical intermediate with CF3I. However, in the case of Li enolate, only the latter path is possible due to the high energy barrier of the elimination of the Li radical. This analysis provides an explanation of the experimental observation that the Li enolate could form the radical cycle efficiently but the Ti ate enolate could not. To make the radical cycle complete, I- has to be extracted from CF3I itself or the radical anion of CF3I. In the case of Li, formation of Li-I bond could be the driving force for the extraction of I- and regeneration of CF3 radical. However, Ti does not give exothermic Ti-I formation and thus regeneration of CF3 radical is less likely. PMID- 17081024 TI - Acyclic or long-bond intermediate in the electron-transfer-catalyzed dimerization of 4-methoxystyrene. AB - The electron-transfer-catalyzed dimerization of 4-methoxystyrene has long been a prototypical reaction for the study of radical cation reactivity. The different possible pathways were explored at the B3LYP/6-31G level of theory. Both [2 + 2] and [4 + 2] cycloadditions proceed via a stepwise pathway, diverging at an acyclic intermediate and interconnected by a vinylcyclobutane-type rearrangement. The experimentally observed stereoselectivity of the cycloaddition was traced to relatively high barriers for isomerization, while the previously described "long bond" intermediate could not be located at the higher level of theory. CPCM calculations show that the highly exothermic [4 + 2] pathway becomes kinetically more favorable in condensed phase. Time-dependent density functional theory calculations indicate that the different possible intermediates have very similar absorption spectra, making the unambiguous assignment of the experimentally observed transient absorption of 500 nm to a given species difficult. PMID- 17081025 TI - Convergent synthesis of complex diketopiperazines derived from pipecolic acid scaffolds and parallel screening against GPCR targets. AB - A convergent approach to highly functionalized diketopiperazines (DKPs) using enantioenriched pipecolic acids is described. Scandium triflate-catalyzed [4 + 2] aza-annulation was employed to produce stereochemically well-defined building blocks. A resin "catch and release" strategy was devised to convert annulation products to pipecolic acid monomers. Complex diketopiperazines were efficiently assembled utilizing one-pot cyclodimerization of pipecolic acids. Massively parallel screening of the complex DKPs against a panel of molecular targets identified novel ligands for a number of G-protein-coupled receptors (GPCRs). PMID- 17081026 TI - Scalable methodology for the catalytic, asymmetric alpha-bromination of acid chlorides. AB - The optimization of a practical, catalytic, asymmetric process for the alpha bromination of acid chlorides to produce synthetically versatile, optically active alpha-bromoesters is reported. A range of products is produced in high enantioselectivity and moderate to good chemical yields with retention of both upon scale-up. The reactions herein are catalyzed by cinchona alkaloid derivatives, with the best performance achieved by the use of a proline cinchona alkaloid conjugate designed in a de novo fashion. PMID- 17081027 TI - Isomerization-free allylic alkylations of terminal pi-allyl palladium complexes. AB - Chelated amino acid ester enolates are excellent nucleophiles for allylic alkylations. With these enolates, even terminal pi-allyl palladium complexes react without significant isomerization. This allows a transfer of the cis-olefin geometry from the substrate into the product. Chiral substrates also show a reasonably good 1,5-induction. PMID- 17081028 TI - Synthesis of cyclic peptides constrained with biarylamine linkers using Buchwald Hartwig C-N coupling. AB - In this paper, we describe the synthesis of conformationally constrained cyclic peptides with biarylamine linkers for peptidomimetics using palladium-catalyzed intramolecular Buchwald-Hartwig C-N coupling. We have prepared a variety of di-, tri-, and tetrapeptides (16-22-membered) in good yields using this reaction. PMID- 17081029 TI - Expanding the C2-symmetric bicyclo[2.2.1]hepta-2,5-diene ligand family: concise synthesis and catalytic activity in rhodium-catalyzed asymmetric addition. AB - New C2-symmetric bicyclo[2.2.1]hepta-2,5-dienes bearing methyl and phenyl substituents at the 2 and 5 positions were prepared enantiomerically pure through a two-step sequence starting from the readily available bicyclo[2.2.1]hepta-2,5 dione. Due to the instability or volatility of these dienes, their isolation was achieved through the formation of the corresponding stable [RhCl(diene)]2 complexes. These chiral rhodium complexes displayed high activity and enantioselectivity (up to 99% ee) in the rhodium-catalyzed 1,4-addition and 1,2 addition of phenylboronic acid to cyclic enones and N-sulfonylimines, respectively. PMID- 17081030 TI - A mild and regioselective method for alpha-bromination of beta-keto esters and 1,3-diketones using bromodimethylsulfonium bromide (BDMS). AB - Bromodimethylsulfonium bromide has been found to be an effective and regioselective reagent for alpha-monobromination of beta-keto esters and 1,3 diketones. A wide variety of beta-keto esters and 1,3-diketones undergo chemoselective alpha-monobromination with excellent yields at 0-5 degrees C or room temperature. The notable advantages of this protocol are no need of chromatographic separation, use of less hazardous reagent than molecular bromine, and no added base, Lewis acid, or other catalyst. PMID- 17081031 TI - An N-heterocyclic carbene-catalyzed [8 + 3] annulation of tropone and enals via homoenolate. AB - A novel protocol for the annulation of tropone to enals involving nucleophilic heterocyclic carbene (NHC) catalyzed homoenolate formation has been developed. Interestingly, the reaction led to bicyclic delta-lactones instead of the expected gamma-spirolactones, presumably by the uncommon [8 + 3] annulation pathway. The strategy works well with a variety of enals. PMID- 17081032 TI - Stereoselective recognition of vicinal diamines with a Zn(II) complex. AB - Zn(II) complex of L (N,N'-bis(2-pyridylmethyl)-N,N'-dimethyl-trans-1,2 diaminocyclohexane) binds chiral vicinal diamines (1,2-diphenylethylenediamine (dpen) and 1,2-diaminocyclohexane (dach)) stereoselectively. Crystallographic studies reveal that the ternary complex has the C2 symmetric cis-alpha topology. 1H NMR shows that the R,R form of the tetradentate zinc complex binds rapidly and reversibly to the R,R form of the diamine over the S,S form with a stereoselectivity of about 5:1. Although the diamine exchange rate is rapid it is slower than the NMR time scale, and distinct signals for the diastereomeric complexes are observed when racemic mixtures of the host and guest molecules are mixed. Origin of stereoselectivity is discussed in terms of steric effects. PMID- 17081033 TI - Facile preparation and functionalization of chiral stabilized ylides from common chiral auxiliaries using triphenyl- phosphoranylideneketene (the Bestmann ylide) and their use in Wittig reactions. AB - Camphor-derived lactams and other related chiral controllers have been found to react with the Bestmann ylide (triphenylphosphoranylideneketene) upon heating in toluene. The resulting parent ylides provide convenient access to a structurally diverse set of chiral stabilized ylides via functionalization. The utility of these chiral ylides for Wittig reactions has been briefly investigated and the effects of alpha-substitution noted. PMID- 17081034 TI - Potassium hydride in paraffin: a useful base for organic synthesis. AB - The preparation of potassium hydride as a 1:1 homogenate with paraffin, termed KH(P), is reported. KH(P), a solid at room temperature, is stable without special handling. On suspension in THF with a phosphonium salt, KH(P) rapidly generates the ylide. Wittig condensation with aromatic, aliphatic, and alpha,beta unsaturated aldehydes proceeds with high Z selectivity. KH(P) should be a generally useful base for organic synthesis. PMID- 17081035 TI - Strategy for the enantioselective synthesis of trans-2,4-disubstituted piperidines: application to the CCR3 antagonist IS811. AB - A strategy for the enantioselective synthesis of trans-2,4-disubstituted piperidines is proposed and applied to the preparation of IS811, a potent CCR3 antagonist. The C2 stereocenter is derived from commercial (R)-epichlorohydrin, while the C4 stereocenter is installed via diastereoselective hydrogenation of an alpha,beta-unsaturated lactone intermediate. Inversion of the original stereocenter via an efficient intramolecular S(N)2 amination affords the piperidine core of IS811. An improved protocol for the lithiation of ethyl propiolate is reported. PMID- 17081036 TI - Preparation of highly substituted 6-arylpurine ribonucleosides by Ni-catalyzed cyclotrimerization. Scope of the reaction. AB - Transition metal complex catalyzed cocyclotrimerization of protected alkynylpurine ribonucleosides 1 with various diynes 2 gave rise to a series of 6 arylpurine nucleosides 3 that were further deprotected to free nucleosides 4. Generally, the best yields of cyclotrimerizations were obtained with a catalytic system Ni(cod)2/2PPh(3). On the other hand, CoBr(PPh(3))3 proved to be a superior catalyst for cyclotrimerization of 1 with dipropargyl ether 2g. In addition, Ni catalysis is also suitable for direct cyclotrimerization of unprotected alkynylpurine ribonucleosides 5 to the corresponding 6-arylpurinylribosides 4. PMID- 17081037 TI - Synthesis and stability of a homologous series of triynol natural products and their analogues. AB - A series of polyyne natural products 1, 13, and 31 and analogues 14, 21, and 22 are synthesized in six steps. The key step is a Fritsch-Buttenberg-Wiechell rearrangement in which a triyne framework is formed from the appropriate dibromoolefin precursor. Terminal conjugated triynes 13 and 14 are obtained as highly unstable products that rapidly decompose under ambient conditions. The stability of triynols increases via either the addition of methylene units (i.e., 6 --> 31 --> 1) or addition of terminal substituents (i.e., 13 --> 21 or 31). PMID- 17081038 TI - First enantiospecific synthesis of (-)-parvifoline and (-)-curcuquinone. AB - The first enantiospecific synthesis of (-)-parvifoline, employing ring-closing metathesis as the key step, and (-)-curcuquinone from naturally occurring (R)-(+) citronellal is described. PMID- 17081039 TI - Thiopyran route to polypropionates: an efficient synthesis of serricornin. AB - The synthesis of serricornin [(4S,6S,7S)-7-hydroxy-4,6-dimethylnonan-3-one], a sex pheromone produced by the female cigarette beetle (Lasioderma serricorne F.), in seven steps from readily available racemic 1,4-dioxa-8-thiaspiro[4.5]decane-6 carboxaldehyde (6) is described. The key steps include enantioselective aldol reaction of 6 with tetrahydrothiopyran-4-one catalyzed by 5-[(2S)-pyrrolidine-2 yl]-1H-tetrazole to fabricate the tetrapropionate skeleton, stereoselective Li(s)Bu(3)BH reduction of the resulting aldol adduct, Barton-McCombie deoxygenation, and Raney nickel desulfurization. PMID- 17081040 TI - New developments in profiling and imaging of proteins from tissue sections by MALDI mass spectrometry. AB - Molecular imaging of tissue by MALDI mass spectrometry is a powerful tool for visualizing the spatial distribution of constituent analytes with high molecular specificity. Although the technique is relatively young, it has already contributed to the understanding of many diverse areas of human health. In recent years, a great many advances in the practice of imaging mass spectrometry have taken place, making the technique more sensitive, robust, and ultimately useful. The purpose of this review is to highlight some of the more recent technological advances that have improved the efficiency of imaging mass spectrometry for clinical applications. Advances in the way MALDI mass spectrometry is integrated with histology, improved methods for automation, and better tools for data analysis are outlined in this review. Refined top-down strategies for the identification and validation of candidate biomarkers found in tissue sections are discussed. A clinical example highlighting the application of these methods to a cohort of clinical samples is described. PMID- 17081041 TI - Proteomic analysis of hypertrophied myocardial protein patterns in renovascularly hypertensive and spontaneously hypertensive rats. AB - The cardiac protein profiles of spontaneously hypertensive and renovascularly hypertensive hypertrophy showed a significant alteration compared with normal hearts. Most proteins with significant modulations in their expressions belong to the category of metabolic and stress-related proteins. Among these proteins, glutathione-S-transferase mu2 and short-chain acyl-CoA dehydrogenase may be two candidate proteins associated with left ventricular hypertrophy in spontaneously hypertensive rats. PMID- 17081042 TI - Detecting differential and correlated protein expression in label-free shotgun proteomics. AB - Recent studies have revealed a relationship between protein abundance and sampling statistics, such as sequence coverage, peptide count, and spectral count, in label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS) shotgun proteomics. The use of sampling statistics offers a promising method of measuring relative protein abundance and detecting differentially expressed or coexpressed proteins. We performed a systematic analysis of various approaches to quantifying differential protein expression in eukaryotic Saccharomyces cerevisiae and prokaryotic Rhodopseudomonas palustris label-free LC-MS/MS data. First, we showed that, among three sampling statistics, the spectral count has the highest technical reproducibility, followed by the less-reproducible peptide count and relatively nonreproducible sequence coverage. Second, we used spectral count statistics to measure differential protein expression in pairwise experiments using five statistical tests: Fisher's exact test, G-test, AC test, t test, and LPE test. Given the S. cerevisiae data set with spiked proteins as a benchmark and the false positive rate as a metric, our evaluation suggested that the Fisher's exact test, G-test, and AC test can be used when the number of replications is limited (one or two), whereas the t-test is useful with three or more replicates available. Third, we generalized the G-test to increase the sensitivity of detecting differential protein expression under multiple experimental conditions. Out of 1622 identified R. palustris proteins in the LC MS/MS experiment, the generalized G-test detected 1119 differentially expressed proteins under six growth conditions. Finally, we studied correlated expression of these 1119 proteins by analyzing pairwise expression correlations and by delineating protein clusters according to expression patterns. Through pairwise expression correlation analysis, we demonstrated that proteins co-located in the same operon were much more strongly coexpressed than those from different operons. Combining cluster analysis with existing protein functional annotations, we identified six protein clusters with known biological significance. In summary, the proposed generalized G-test using spectral count sampling statistics is a viable methodology for robust quantification of relative protein abundance and for sensitive detection of biologically significant differential protein expression under multiple experimental conditions in label-free shotgun proteomics. PMID- 17081043 TI - Proteome resolution by two-dimensional gel electrophoresis varies with the commercial source of IPG strips. AB - By facilitating reproducible first dimension separations, commercial immobilized pH gradient (IPG) strips enable high throughput and high-resolution proteomic analyses using two-dimensional gel electrophoresis (2DE). Amersham, Biorad, Invitrogen, and Sigma all market linear pH 3-10 IPG strips. We have applied optimized 2DE protocols with both membrane and soluble brain protein extracts to critically evaluate all four products. Resolved protein spots were quantitatively evaluated after carrying out these protocols using IPG strips from the four companies. Biorad and Amersham IPG strips resolved a high number of membrane and soluble proteins, respectively. Furthermore, Amersham IPG strips eluted the largest amount of protein into the second dimension gels and had the most protein remaining in the strip after 2DE. Biorad and Amersham IPG strips maintained a consistent linear pH 3-10 gradient, whereas those from Invitrogen appeared nonlinear or "compressed" within the central pH region. The gradient range within Sigma IPG strips appeared to be slightly less than pH 3-10, due to one extended pH unit within the gradient. Overall, all four commercially available IPG strips have the ability to resolve both membrane and soluble brain proteomes. The difference is that Amersham and Biorad do so more consistently and with better spot resolution. It appears that the physical/chemical nature of commercially available IPG strips can vary considerably, leading to marked differences in subsequent protein resolution in 2DE. These differences likely reflect variations in the uptake of proteins into the strips, and differences in the focusing and elution of proteins from the first to the second dimension. These differences would appear, in part, to underlie some inter-lab variations in the effective resolution of proteomes. PMID- 17081044 TI - Multiplexed mass spectrometric immunoassay in biomarker research: a novel approach to the determination of a myocardial infarct. AB - Reported here is the development of a multiplexed mass spectrometric immunoassay (MSIA) for the detection of myocardial infarction (MI). The assay is the product of a study that systematically progresses from biomarker discovery--to identification and verification--to assay design, data analysis, and statistical challenge. During targeted population proteomics investigations, two novel biomarkers, serum amyloid A1alpha and S-sulfated transthyretin, were found to be responsive to MI. These putative markers were subsequently screened in larger cohorts of individuals to verify their responsiveness toward MI. Upon verification, a multiplexed assay was designed that was capable of simultaneously monitoring the new markers plus a previously established MI-marker (myoglobin). The multiplexed MSIA was applied to two 96-sample sets comprised of 48-MI/48 healthy and 19-MI/77-healthy, which served as training and case cohorts, respectively. Data evaluation using either preset reference levels or multivariate analysis exhibited sensitivities and specificities of >97%. These findings illustrate the importance of using systematic approaches in clinical proteomics to discover biomarkers and produce high-performance assays relevant to disease. PMID- 17081045 TI - Probing the structure of the Caulobacter crescentus ribosome with chemical labeling and mass spectrometry. AB - The ribosomal proteins of Caulobacter crescentus were amidinated before and after disassembly of the organelle and the results analyzed by mass spectrometry. Comparison with structural information from previous X-ray crystal studies of other bacterial ribosomes provides insight about the C. crescentus ribosome. In total, 47 of the 54 proteins present in the ribosome of C. crescentus were detected after labeling. The extent of derivatization for each protein is strongly dependent on the solvent accessibility of its target residues. Proteins of the ribosome stalk, which are known to be largely solvent-accessible, were labeled quite extensively. In striking contrast, other proteins that are known to be highly shielded in their subunits were labeled at very few of their potential sites. Furthermore, evidence that protein L12 binds to the ribosome via its N terminal domain is consistent with previous findings. PMID- 17081046 TI - Declining plasma fibrinogen alpha fragment identifies HER2-positive breast cancer patients and reverts to normal levels after surgery. AB - Breast cancer is the most common nonskin malignancy affecting women. Currently, no simple, blood-based diagnostic test exists to complement radiological screening and increase sensitivity of detection. To screen plasma specimens and identify biomarkers that detect HER2-positive breast cancer, automated robotic sample processing followed by surface-enhanced laser desorption ionization time of-flight (SELDI-TOF) mass spectroscopy was used. Multiple statistical algorithms were used to select biomarkers that segregate cancer patients versus controls and produced average CV rates ranging from 20% to 29%. A set of seven biomarkers were validated on an independent test data set and achieved the best error rate of 19.1%. A permutation test indicated a p-value for CV error less than 0.002. Moreover, a ROC curve using these biomarkers achieved an area-under-the-curve value of 0.95 on an independent test data set. The marker responsible for most of the resolving power was identified as a fragment of Fibrinogen Alpha (FGA) encompassing residues 605-629. This marker was present at lower levels in cancer patients as compared to controls. The importance of this biomarker was validated in a longitudinal study comparing pre- and post-operative levels and was shown to revert to normal levels after surgery. This fragment may serve as a useful diagnostic and treatment-monitoring marker. PMID- 17081047 TI - Comparison of hydroxylated print additives on antibody microarray performance. AB - Various hydroxylated additives were added to antibody print buffers at different concentrations to stabilize printed antibodies during normal array spot desiccation on commercial polymer-coated microarray slides. Polyvinyl alcohol addition to print buffers produced the most regular spot morphologies, homogeneous intra-spot antibody distribution, uniform fluorescence intensity, and improved analyte capture activity, maintained up to 1 month at 4 degrees C for capturing model analytes, anti-human IL-1beta, IL-4, and TNFalpha, on these microarraying slides. PMID- 17081048 TI - Time-dependent plasma protein changes in streptozotocin-induced diabetic rats before and after fungal polysaccharide treatments. AB - Previous studies about protein modulation with chemically induced models of diabetes in animals have yielded conflicting results, in that many investigators have reported different regulation patterns for the same proteins. Therefore, it is reasonable to determine biomarkers for prognosis and diagnosis of diabetes with time profiling for the candidate proteins. In this regard, we examined the influence of hypoglycemic fungal polysaccharides (EPS) on the time-dependent plasma protein alterations in streptozotocin-induced diabetic rats. The 2-DE analysis of rat plasma demonstrated that about 50 proteins from about 900 visualized spots were found to be differentially regulated, of which 20 spots were identified as principal diabetes-associated proteins. The results of time profiling revealed that most of the identified proteins showed significant alterations in a time-dependent manner during 14 days, with notable trends. Nine out of the twenty proteins displayed very similar time profiles between normal healthy and EPS-treated diabetic rats. Interestingly, the altered profiles of several proteins by diabetes induction almost returned to control levels after EPS treatments. In particular, we found a clear distinction in differential expression of oxidative stress proteins (ceruloplasmin and transferrin) and lipid metabolism related proteins (Apo A-I, Apo A-IV, and Apo E) in the STZ-induced diabetic rats. The data presented here have identified and characterized the time dependent changes in plasma proteins associated with EPS treatment in STZ-induced diabetic rats, thereby leading to the discovery of early-response and late response biomarkers in diabetic and EPS-treated states. PMID- 17081049 TI - Toward plasma proteome profiling with ion mobility-mass spectrometry. AB - Differential, functional, and mapping proteomic analyses of complex biological mixtures suffer from a lack of component resolution. Here we describe the application of ion mobility-mass spectrometry (IMS-MS) to this problem. With this approach, components that are separated by liquid chromatography are dispersed based on differences in their mobilities through a buffer gas prior to being analyzed by MS. The inclusion of the gas-phase dispersion provides more than an order of magnitude enhancement in component resolution at no cost to data acquisition time. Additionally, the mobility separation often removes high abundance species from spectral regions containing low-abundance species, effectively increasing measurement sensitivity and dynamic range. Finally, collision-induced dissociation of all ions can be recorded in a single experimental sequence while conventional MS methods sequentially select precursors. The approach is demonstrated in a single, rapid (3.3 h) analysis of a plasma digest sample where abundant proteins have not been removed. Protein database searches have yielded 731 high confidence peptide assignments corresponding to 438 unique proteins. Results have been compiled into an initial analytical map to be used -after further augmentation and refinement- for comparative plasma profiling studies. PMID- 17081050 TI - Disorder and sequence repeats in hub proteins and their implications for network evolution. AB - Protein interaction networks display approximate scale-free topology, in which hub proteins that interact with a large number of other proteins determine the overall organization of the network. In this study, we aim to determine whether hubs are distinguishable from other networked proteins by specific sequence features. Proteins of different connectednesses were compared in the interaction networks of Saccharomyces cerevisiae, Drosophila melanogaster, Caenorhabditis elegans, and Homo sapienswith respect to the distribution of predicted structural disorder, sequence repeats, low complexity regions, and chain length. Highly connected proteins ("hub proteins") contained significantly more of, and greater proportion of, these sequence features and tended to be longer overall as compared to less connected proteins. These sequence features provide two different functional means for realizing multiple interactions: (1) extended interaction surface and (2) flexibility and adaptability, providing a mechanism for the same region to bind distinct partners. Our view contradicts the prevailing view that scaling in protein interactomes arose from gene duplication and preferential attachment of equivalent proteins. We propose an alternative evolutionary network specialization process, in which certain components of the protein interactome improved their fitness for binding by becoming longer or accruing regions of disorder and/or internal repeats and have therefore become specialized in network organization. PMID- 17081051 TI - Proteomic analysis of plasma membrane from hypoxia-adapted malignant melanoma. AB - Hypoxic conditions often persist within poorly vascularized tumors. At the cellular level constitutive activation of transcriptional regulators of the hypoxic response leads to the emergence of clones with aggressive phenotypes. The primary interface between the cell and the hypoxic environment is the plasma membrane. A detailed investigation of this organelle is expected to yield further targets for therapeutic perturbation of the response to hypoxia. In the present study, quantitative proteomic analysis of plasma membrane from hypoxia-adapted murine B16F10 melanoma was performed using differential 16O/18O stable isotopic labeling and multidimensional liquid chromatography-tandem mass spectrometry. The analysis resulted in the identification of 24,853 tryptic peptides, providing quantitative information for 2,433 proteins. For a subset of plasma membrane and secreted proteins, quantitative RT-PCR was used to gain further insight into the genomic regulatory events underlying the response to hypoxia. Consistent increases at the proteomic and transcriptomic levels were observed for aminopeptidase N (CD13), carbonic anhydrase IX, potassium-transporting ATPase, matrix metalloproteinase 9, and stromal cell derived factor I (SDF-1). Antibody based analysis of a panel of human melanoma cell lines confirmed that CD13 and SDF-1 were consistently upregulated during hypoxia. This study provides the basis for the discovery of novel hypoxia-induced membrane proteins. PMID- 17081052 TI - Biomarker candidate identification in Yersinia pestis using organism-wide semiquantitative proteomics. AB - The accurate mass and time tag mass spectrometry method and clustering analysis were used to compare the abundance change of 992 Yersinia pestis proteins under four contrasting growth conditions (26 and 37 degrees C, with or without Ca2+) that mimicked growth states in either a flea vector or mammalian host. Eighty nine proteins were observed to have similar abundance change profiles to 29 known virulence associated proteins, providing identification of additional biomarker candidates. Eighty-seven hypothetical proteins, which clustered into 5 distinct clusters of like-protein abundance change, were identified as unique biomarkers related specifically to growth condition. PMID- 17081053 TI - Performance evaluation of existing de novo sequencing algorithms. AB - Two methods have been developed for protein identification from tandem mass spectra: database searching and de novo sequencing. De novo sequencing identifies peptide directly from tandem mass spectra. Among many proposed algorithms, we evaluated the performance of the five de novo sequencing algorithms, AUDENS, Lutefisk, NovoHMM, PepNovo, and PEAKS. Our evaluation methods are based on calculation of relative sequence distance (RSD), algorithm sensitivity, and spectrum quality. We found that de novo sequencing algorithms have different performance in analyzing QSTAR and LCQ mass spectrometer data, but in general, perform better in analyzing QSTAR data than LCQ data. For the QSTAR data, the performance order of the five algorithms is PEAKS > Lutefisk, PepNovo > AUDENS, NovoHMM. The performance of PEAKS, Lutefisk, and PepNovo strongly depends on the spectrum quality and increases with an increase of spectrum quality. However, AUDENS and NovoHMM are not sensitive to the spectrum quality. Compared with other four algorithms, PEAKS has the best sensitivity and also has the best performance in the entire range of spectrum quality. For the LCQ data, the performance order is NovoHMM > PepNovo, PEAKS > Lutefisk > AUDENS. NovoHMM has the best sensitivity, and its performance is the best in the entire range of spectrum quality. But the overall performance of NovoHMM is not significantly different from the performance of PEAKS and PepNovo. AUDENS does not give a good performance in analyzing either QSTAR and LCQ data. PMID- 17081054 TI - Global profiling of surface plasma membrane proteome of oviductal epithelial cells. AB - In mammalian reproduction, many important events occur within the female reproductive tract, especially within the oviduct. These include transport and final maturation of the female and male gametes, fertilization, embryonic development, and transport of the embryo to the uterus. The plasma membrane molecules of oviductal epithelia that are in direct contact with gametes and embryo(s) and potentially mediate these processes are poorly characterized, and their function is poorly understood. Defining the oviductal cell surface proteome could provide a better understanding of the basis of reproductive processes taking place within the oviduct. We aimed to provide a detailed profile of the surface plasma membrane proteome of the oviductal epithelium by biotinylation of proteins at the cell surface, followed by highly specific purification of these proteins using avidin. This approach for enrichment of oviductal cell surface proteome was validated by immunohistochemistry, gel electrophoresis, and western blot analysis experiments. The enriched molecules were identified using two different technologies: (i) the combination of 2D gel electrophoresis with mass spectrometry and (ii) 1D gel electrophoresis with mass spectrometry (a modified multidimensional protein identification technology (MudPIT) technique). The number of proteins identified using the MudPIT approach was approximately 7 times the number of proteins identified by 2D gel electrophoresis using the same samples (40 versus 276, respectively). Some of the proteins found at the surface of oviductal cells had previously been reported as present in the oviduct and to have known functions in relation to reproductive processes. The other category of proteins that were highly represented in the oviductal surface proteome were various members of the family of heat-shock proteins. To the best of our knowledge, this is the first comprehensive study to identify and characterize proteins at the surface of the epithelium of the mammalian oviduct. PMID- 17081055 TI - Serial changes in urinary proteome profile of membranous nephropathy: implications for pathophysiology and biomarker discovery. AB - Membranous nephropathy is one of the most common causes of primary glomerular diseases worldwide. The present study adopted a gel-based proteomics approach to better understand the pathophysiology and define biomarker candidates of human membranous nephropathy using an animal model of passive Heymann nephritis (PHN). Clinical characteristics of Sprague-Dawley rats injected with rabbit anti-Fx1A antiserum mimicked those of human membranous nephropathy. Serial urine samples were collected at Days 0, 10, 20, 30, 40, and 50 after the injection with anti Fx1A (number of rats = 6; total number of gels = 36). Urinary proteome profiles were examined using 2D-PAGE and SYPRO Ruby staining. Quantitative intensity analysis and ANOVA with Tukey post-hoc multiple comparisons revealed 37 differentially expressed proteins among 6 different time-points. These altered proteins were successfully identified by MALDI-TOF MS and classified into 6 categories: (i) proteins with decreased urinary excretion during PHN; (ii) proteins with increased urinary excretion during PHN; (iii) proteins with increased urinary excretion during PHN, but which finally returned to basal levels; (iv) proteins with increased urinary excretion during PHN, but which finally declined below basal levels; (v) proteins with undetectable levels in the urine during PHN; and (vi) proteins that were detectable in the urine only during PHN. Most of these altered proteins have functional significance in signaling pathways, glomerular trafficking, and controlling the glomerular permeability. The ones in categories (v) and (vi) may serve as biomarkers for detecting or monitoring membranous nephropathy. After normalization of the data with 24-h urine creatinine excretion, changes in 34 of initially 37 differentially expressed proteins remained statistically significant. These data underscore the significant impact of urinary proteomics in unraveling disease pathophysiology and biomarker discovery. PMID- 17081056 TI - Expressed peptide tags: an additional layer of data for genome annotation. AB - While genome sequencing is becoming ever more routine, genome annotation remains a challenging process. Identification of the coding sequences within the genomic milieu presents a tremendous challenge, especially for eukaryotes with their complex gene architectures. Here, we present a method to assist the annotation process through the use of proteomic data and bioinformatics. Mass spectra of digested protein preparations of the organism of interest were acquired and searched against a protein database created by a six-frame translation of the genome. The identified peptides were mapped back to the genome, compared to the current annotation, and then categorized as supporting or extending the current genome annotation. We named the classified peptides Expressed Peptide Tags (EPTs). The well-annotated bacterium Rhodopseudomonas palustris was used as a control for the method and showed a high degree of correlation between EPT mapping and the current annotation, with 86% of the EPTs confirming existing gene calls and less than 1% of the EPTs expanding on the current annotation. The eukaryotic plant pathogens Phytophthora ramorum and Phytophthora sojae, whose genomes have been recently sequenced and are much less well-annotated, were also subjected to this method. A series of algorithmic steps were taken to increase the confidence of EPT identification for these organisms, including generation of smaller subdatabases to be searched against, and definition of EPT criteria that accommodates the more complex eukaryotic gene architecture. As expected, the analysis of the Phytophthora species showed less correlation between EPT mapping and their current annotation. While approximately 76% of Phytophthora EPTs supported the current annotation, a portion of them (7.7% and 12.9% for P. ramorum and P. sojae, respectively) suggested modification to current gene calls or identified novel genes that were missed by the current genome annotation of these organisms. PMID- 17081057 TI - Characterizing the reproducibility of a protein profiling method for the analysis of mouse bronchoalveolar lavage fluid. AB - The detection of biomarkers in biological fluids has been advanced by the introduction of mass spectrometry screening methods such as matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS), which enables the detection of the presence and the molecular mass of proteins in unfractionated mixtures. The generation of reproducible mass spectra over the course of an experiment is vital in obtaining data in which differences in protein profiles between diseased and healthy states can be assessed correctly. We have developed a protocol to automate the collection of protein profiling data from a large number of samples using MALDI-TOFMS, and we used these samples to characterize the technical reproducibility of the method. This protocol has been used for the analysis of proteins found in bronchoalveolar lavage fluid samples from mice with the ultimate goal of enabling the discovery of differential expression patterns predictive of the development of chronic obstructive pulmonary disease. Samples were purified using magnetic bead-based technology and analyzed on an AnchorChip target plate. Our results demonstrate that the number of peaks detected reproducibly decreases significantly as sample size increases, which motivates the need for technical replicates to be explicitly included in the analysis of MALDI-TOF-based protein profiling studies. PMID- 17081058 TI - Mass spectrometry-based glycoproteomic approach involving lysine derivatization for structural characterization of recombinant human erythropoietin. AB - Lysine-containing peptides comprising glycosylation sites derived from recombinant human erythropoietin (rHuEPO) by trypsin or Lys-C and PNGase F dual digestion were derivatized with 2-methoxy-4,5-dihydro-1H-imidazole and its deuterated analogues. In the same reaction, under reducing conditions (beta mercaptoethanol), cysteines were converted into methyl-cysteines and lysines into Lys-4,5-dihydro-1H-imidazole. Both modifications on cysteines and lysines simplified the CID-MS/MS spectra, while preserving the structural information by yielding y-series ions and improved the mass spectral signal intensity up to 25 times. Moreover, by this approach, the N-glycan occupation sites were unambiguously determined. O-Glycosylation sites as well as O-glycan structures were determined by a LC-MS/MS experiment carried out on dually digested rHuEPO. N Glycan mixture purified on a graphitized carbon column using a newly developed method that extracted only sialylated carbohydrates was analyzed first using MALDI-TOF in negative linear ion mode with low mass accuracy but without interferences and metastabile ions and then a reflectron with high mass accuracy. After defining the precursor ions, we performed the nanoESI QTOF MS/MS analysis on N-glycans, mainly targeting the distinction between carbohydrates with sialylated antennae and those lacking sialic acid moieties. PMID- 17081059 TI - Allosteric modulation of dopamine D2 receptors by homocysteine. AB - It has been suggested that L-DOPA-induced hyperhomocysteinemia can increase the risk of stroke, heart disease, and dementia and is an additional pathogenetic factor involved in the progression of Parkinson's disease. In Chinese hamster ovary (CHO) cells stably cotransfected with adenosine A(2A) and dopamine D2 receptors, homocysteine selectively decreased the ability of D2 receptor stimulation to internalize adenosine A(2A)-dopamine D2 receptor complexes. Radioligand-binding experiments in the same cell line demonstrated that homocysteine acts as an allosteric D2 receptor antagonist, by selectively reducing the affinity of D2 receptors for agonists but not for antagonists. Mass spectrometric analysis showed that, by means of an arginine (Arg)-thiol electrostatic interaction, homocysteine forms noncovalent complexes with the two Arg-rich epitopes of the third intracellular loop of the D2 receptor, one of them involved in A(2A)-D2 receptor heteromerization. However, homocysteine was unable to prevent or disrupt A(2A)-D2 receptor heteromerization, as demonstrated with Fluorescence Resonance Energy Transfer (FRET) experiments in stably cotransfected HEK cells. The present results could have implications for Parkinson's disease. PMID- 17081060 TI - Identification of ethylene-mediated protein changes during nodulation in Medicago truncatula using proteome analysis. AB - Ethylene has been hypothesised to be a regulator of root nodule development in legumes, but its molecular mechanisms of action remain unclear. The skl mutant is an ethylene-insensitive legume mutant showing a hypernodulation phenotype when inoculated with its symbiont Sinorhizobium meliloti. We used the skl mutant to study the ethylene-mediated protein changes during nodule development in Medicago truncatula. We compared the root proteome of the skl mutant to its wild-type in response to the ethylene precursor aminocyclopropane carboxylic acid (ACC) to study ethylene-mediated protein expression in root tissues. We then compared the proteome of skl roots to its wild-type after Sinorhizobium inoculation to identify differentially displayed proteins during nodule development at 1 and 3 days post inoculation (dpi). Six proteins (pprg-2, Kunitz proteinase inhibitor, and ACC oxidase isoforms) were down-regulated in skl roots, while three protein spots were up-regulated (trypsin inhibitor, albumin 2, and CPRD49). ACC induced stress-related proteins in wild-type roots, such as pprg-2, ACC oxidase, proteinase inhibitor, ascorbate peroxidase, and heat-shock proteins. However, the expression of stress-related proteins such as pprg-2, Kunitz proteinase inhibitor, and ACC oxidase, was down-regulated in inoculated skl roots. We hypothesize that during early nodule development, the plant induces ethylene mediated stress responses to limit nodule numbers. When a mutant defective in ethylene signaling, such as skl, is inoculated with rhizobia, the plant stress response is reduced, resulting in increased nodule numbers. PMID- 17081061 TI - A soluble 3D LC/MS/MS proteome of the filamentous cyanobacterium Nostoc punctiforme. AB - Nostoc punctiforme is an oxygenic photoautotrophic cyanobacterium with multiple developmental states, which can form nitrogen-fixing symbioses with a variety of terrestrial plants. 3D LC/MS/MS shotgun peptide sequencing was used to analyze the proteome when N. punctiforme is grown in continuous moderate light with ammonia as the nitrogen source. The soluble proteome includes 1575 proteins, 50% of which can be assigned to core metabolic and transport functions. Another 39% are assigned to proteins with no known function, a substantially higher fraction than in the Escherichia coli proteome. Many expressed proteins protect against oxidative and light stress. Seventy-one sensor histidine kinases, response regulators, and serine/threonine kinases, individually and as hybrid, multidomain proteins, were identified, reflecting a substantial capacity to sense and respond to environmental change. Proteins encoded by each of the five N. punctiforme plasmids were identified, as were 10 transposases, reflecting the plasticity of the N. punctiforme genome. This core proteome sets the stage for comparison with that of other developmental states. PMID- 17081062 TI - Enrichment and identification of integral membrane proteins from barley aleurone layers by reversed-phase chromatography, SDS-PAGE, and LC-MS/MS. AB - The plasma membrane of the cereal aleurone layer is the site of perception of germination signals and release of enzymes to the starchy endosperm. Analysis of membrane proteins is challenging due to their hydrophobicity and low abundance; thus, little is known about the membrane proteins involved in seed germination. A membrane fraction highly enriched for the plasma membrane H+-ATPase was prepared from barley aleurone layers by aqueous two-phase partitioning. Because detergent and salt washes did not efficiently remove soluble proteins from the membrane preparations, an alternative procedure was developed, comprising batch reversed phase chromatography with stepwise elution of hydrophobic proteins by 2-propanol. Proteins in the most hydrophobic fraction were separated by SDS-PAGE and identified by LC-MS/MS and barley EST sequence database search. The method was efficient for enrichment of integral membrane proteins with relatively low levels of soluble contaminating proteins. Forty-six proteins associated with barley aleurone plasma membranes were identified, including proteins with more than 10 transmembrane domains. Among the identified proteins were two new isoforms of the plasma membrane H+-ATPase, two proteins possibly involved in ion-channel regulation, and two proteins of unknown function. This represents the first analysis of membrane proteins involved in seed germination using a proteomics approach. PMID- 17081063 TI - Enrichment of phosphopeptides by Fe3+-immobilized mesoporous nanoparticles of MCM 41 for MALDI and nano-LC-MS/MS analysis. AB - Fe3+-immobilized mesoporous molecular sieves MCM-41 with particle size of ca. 600 nm and pore size of ca. 3 nm is synthesized and applied to selectively trap and separate phosphopeptides from tryptic digest of proteins. For the capture of phosphopeptides, typically 10 microL of tryptic digest solution was first diluted to 1 mL by solution of ACN/0.1% TFA (50:50, v/v) and incubated with 10 microL of 0.1% acetic acid dispersed Fe3+-immobilized MCM-41 for 1 h under vibration. Fe3+ immobilized MCM-41 with trapped phosphopeptides was separated by centrifugation. The deposition was first washed with a volume of 300 microL of solution containing 100 mM NaCl in ACN/0.1% TFA (50:50, v/v) and followed by a volume of 300 microL of solution of 0.1% acetic acid to remove nonspecifically bound peptides. The nanoparticles with trapped phosphopeptides are mixed with 2,5 dihydroxybenzoic acid (2,5-DHB) and deposited onto the target for analysis by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). It was found that phosphopeptides from tryptic digest of alpha-casein and beta-casein are effectively and specifically trapped on Fe3+-immobilized MCM-41 with few peptides nonspecifically adsorbed. After the extraction by Fe3+-immobilized MCM 41, the suppression to the detection of phosphopeptides caused by abundant nonphosphopeptides from tryptic digest is effectively eliminated, and the detection of phosphopeptides by MALDI is greatly enhanced with the value of signal-to-noise (S/N) increased by more than an order of magnitude. It is demonstrated that the mechanism of the adsorption of phosphopeptides on Fe3+ immobilized MCM-41 is based on the interaction between the Fe3+ and the phosphate group. Finally, Fe3+-immobilized MCM-41 is applied to extract phosphopeptides from tryptic digest of the lysate of mouse liver for phosphoproteome analysis by nano-LC-MS/MS. PMID- 17081064 TI - Comparative proteome analysis of fractions enriched for membrane-associated proteins from Francisella tularensis subsp. tularensis and F. tularensis subsp. holarctica strains. AB - The facultative intracellular pathogen Francisella tularensis is the causative agent of the serious infectious disease tularemia. Despite intensive research, the virulence factors and pathogenetic mechanisms remain largely unknown. To identify novel putative virulence factors, we carried out a comparative proteome analysis of fractions enriched for membrane-associated proteins isolated from the highly virulent subspecies tularensis strain SCHU S4 and three representatives of subspecies holarctica of different virulence including the live vaccine strain. We identified six proteins uniquely expressed and four proteins expressed at significantly higher levels by SCHU S4 compared to the ssp. holarctica strains. Four other protein spots represented mass and charge variants and seven spots were charge variants of proteins occurring in the ssp. holarctica strains. The genes encoding proteins of particular interest were examined by sequencing in order to confirm and explain the findings of the proteome analysis. Our studies suggest that the subspecies tularensis-specific proteins represent novel potential virulence factors. PMID- 17081065 TI - Proteomic and bioinformatic characterization of the biogenesis and function of melanosomes. AB - Melanin, which is responsible for virtually all visible skin, hair, and eye pigmentation in humans, is synthesized, deposited, and distributed in subcellular organelles termed melanosomes. A comprehensive determination of the protein composition of this organelle has been obstructed by the melanin present. Here, we report a novel method of removing melanin that includes in-solution digestion and immobilized metal affinity chromatography (IMAC). Together with in-gel digestion, this method has allowed us to characterize melanosome proteomes at various developmental stages by tandem mass spectrometry. Comparative profiling and functional characterization of the melanosome proteomes identified approximately 1500 proteins in melanosomes of all stages, with approximately 600 in any given stage. These proteins include 16 homologous to mouse coat color genes and many associated with human pigmentary diseases. Approximately 100 proteins shared by melanosomes from pigmented and nonpigmented melanocytes define the essential melanosome proteome. Proteins validated by confirming their intracellular localization include PEDF (pigment-epithelium derived factor) and SLC24A5 (sodium/potassium/calcium exchanger 5, NCKX5). The sharing of proteins between melanosomes and other lysosome-related organelles suggests a common evolutionary origin. This work represents a model for the study of the biogenesis of lysosome-related organelles. PMID- 17081066 TI - Multidimensional proteomic analysis of the soluble subproteome of the emerging nosocomial pathogen Ochrobactrum anthropi. AB - We report the first large-scale gel-free proteomic analysis of the soluble subproteome of the emerging pathogen Ochrobactrum anthropi. Utilizing our robust offline multidimensional protein identification protocol, a total of 57 280 peptides were initially identified utilizing automated MS/MS analysis software. We describe our investigation of the heuristic protein validation tool PROVALT and demonstrate its ability to increase the speed and accuracy of the curation process of large-scale proteomic datasets. PROVALT reduced our peptide list to 8517 identified peptides and further manual curation of these peptides led to a final list of 984 uniquely identified peptides that resulted in the positive identification of 249 proteins. These identified proteins were functionally classified and physiochemically characterized. A variety of typical "housekeeping" functions identified within the proteome included nucleic acid, amino and fatty acid anabolism and catabolism, glycolysis, TCA cycle, and pyruvate and selenoamino acid metabolism. In addition, a number of potential virulence factors of relevance to both plant and human disease were identified. PMID- 17081067 TI - Sepsis plasma protein profiling with immunodepletion, three-dimensional liquid chromatography tandem mass spectrometry, and spectrum counting. AB - Sepsis is a systemic, often fatal inflammatory response whose biochemical pathways are not fully understood and with no single biomarker capable of its reliable prediction. Increased interest in protein profiling to reveal fundamental biochemical events as well as disease diagnosis has grown considerably, largely due to advances in mass spectrometry and related front-end technologies. In this study, patients with sepsis and systemic inflammatory response syndrome (SIRS) were examined using plasma protein profiling following immunodepletion treatment to remove the most abundant proteins, serum albumin, transferrin, haptoglobin, anti-trypsin, IgG, and IgA. These proteins cause significant signal suppression, and their removal allows for lower abundance proteins to be examined through improved ion signal. Analyses after immunodepletion were performed using 3-dimensional reverse phase/strong cation exchange/reverse phase liquid chromatography with electrospray ion trap mass spectrometry (3D LC-MS/MS) and spectrum counting for comparative quantitation. The results revealed a major theme in immune system activity, including activation of the complement and coagulation pathways. Additionally, lipid transport may prove to be important in distinguishing sepsis from SIRS. Specifically, significant multi-fold changes were observed in 10 proteins and are now being investigated for the early diagnosis of sepsis. PMID- 17081068 TI - An initial proteomic analysis of human preterm labor: placental membranes. AB - Human preterm labor (PL) is the single most significant problem in modern Obstetrics and Gynecology, affecting approximately 10% of pregnancies worldwide, constituting the leading cause of perinatal mortality and morbidity, and contributing significantly to chronic childhood disease. Currently, our molecular understanding of PL remains staggeringly inadequate to reliably diagnose or rationally intervene in PL events; several molecular alterations have been implicated in PL, but these have proven of limited value as diagnostic/prognostic markers. The majority of PL events remain spontaneous and unpredictable: critical care emergencies. Here, we apply functional proteomics to dissect molecular mechanisms of human PL. Human placental tissue was collected in clearly differentiated cases of preterm and term labor. Highly refined two-dimensional gel electrophoresis (2DE) was used for protein separation, coupled with automated differential gel image analysis to compare the resulting proteomic maps. For this initial study, only the most important protein differences were selected for further analysis, that is, proteins that were unique to one sample, and absent from the other, with 100% reproducibility across the sample population. In total, 11 such proteins were identified by tandem mass spectrometry, falling into three distinct functional classes: structural/cytoskeletal components, ER lumenal proteins with enzymatic or chaperone functions, and proteins with anticoagulant properties. These expression changes form the groundwork for further molecular investigation of this devastating medical condition. This approach therefore holds the potential not only to define the underlying molecular components, but also to identify novel diagnostic tools and targets for rational drug intervention. PMID- 17081069 TI - Zinc through the three domains of life. AB - Zinc is one of the metal ions essential for life, as it is required for the proper functioning of a large number of proteins. Despite its importance, the annotation of zinc-binding proteins in gene banks or protein domain databases still has significant room for improvement. In the present work, we compiled a list of known zinc-binding protein domains and of known zinc-binding sequence motifs (zinc-binding patterns), and then used them jointly to analyze the proteome of 57 different organisms to obtain an overview of zinc usage by archaeal, bacterial, and eukaryotic organisms. Zinc-binding proteins are an abundant fraction of these proteomes, ranging between 4% and 10%. The number of zinc-binding proteins correlates linearly with the total number of proteins encoded by the genome of an organism, but the proportionality constant of Eukaryota (8.8%) is significantly higher than that observed in Bacteria and Archaea (from 5% to 6%). Most of this enrichment is due to the larger portfolio of regulatory proteins in Eukaryota. PMID- 17081070 TI - Interactive feature finding in liquid chromatography mass spectrometry data. AB - We propose a method for finding features in liquid chromatography mass spectrometry data that is based on the isotopic pattern of peaks. Our interactive approach to feature finding is carried out across many samples simultaneously and aligns features concurrently. Our scale-independent approach prioritises potential features and is easily adaptable to look for features of a particular mass and charge, paired features in isotopically labeled samples, or differentially expressed features. We demonstrate this by identifying features from normal human adult plasma. We highlight properties of plasma data that illustrate the need to visually check the quality of features found prior to further statistical analysis. PMID- 17081071 TI - Capillary array reversed-phase liquid chromatography-based multidimensional separation system coupled with MALDI-TOF-TOF-MS detection for high-throughput proteome analysis. AB - A high-throughput on-line capillary array-based two-dimensional liquid chromatography (2D-LC) system coupled with MALDI-TOF-TOF-MS proteomics analyzer for comprehensive proteomic analyses has been developed, in which one capillary strong-cation exchange (SCX) chromatographic column was used as the first separation dimension and 18 parallel capillary reversed-phase liquid chromatographic (RPLC) columns were integrated as the second separation dimension. Peptides bound to the SCX phase were "stepped" off using multiple salt pulses followed by sequentially loading of each subset of peptides onto the corresponding precolumns. After salt fractionation, by directing identically split solvent-gradient flows into 18 channels, peptide fractions were concurrently back-flushed from the precolumns and separated simultaneously with 18 capillary RP columns. LC effluents were directly deposited onto the MALDI target plates through an array of capillary tips at a 15-s interval, and then alpha-cyano-4-hydroxycinnamic acid (CHCA) matrix solution was added to each sample spot for subsequent MALDI experiments. This new system allows an 18-fold increase in throughput compared with serial-based 2D-LC system. The high efficiency of the overall system was demonstrated by the analysis of a tryptic digest of proteins extracted from normal human liver tissue. A total of 462 proteins was identified, which proved the system's promising potential for high throughput analysis and application in proteomics. PMID- 17081072 TI - Characterization of the outer membrane protein profile from disease-related Helicobacter pylori isolates by subcellular fractionation and nano-LC FT-ICR MS analysis. AB - Because of the important role of membrane proteins in adhesion, invasion, and intracellular survival of pathogens in the host, membrane proteins are of potential interest in the search for drug targets or biomarkers. We have established a mass spectrometry-based method that allows characterization of the outer membrane protein (OMP) profile of clinical isolates from of the human gastric pathogen Helicobacter pylori. Subcellular fractionation and one dimensional gel electrophoresis (1D-GE) analysis was combined with nano-liquid chromatography Fourier transform-ion cyclotron resonance mass spectrometry (nano LC FT-ICR MS) and tandem mass spectrometry (MS/MS) analysis of fifteen H. pylori strains associated either with duodenal ulcers, gastric cancer, or isolated from asymptomatic H. pylori infected carriers. Over 60 unique membrane or membrane associated proteins, including 30 of the 33 theoretically predicted OMPs, were identified from the strains. Several membrane proteins, including Omp11 and BabA, were found to be expressed by all strains. In the search for clinical markers we found that Omp26 was expressed by all disease-related strains but was only present in one out of five strains from asymptomatic carriers, which makes Omp26 a potential target for further investigation in the search for proteins unique to disease-related H. pylori strains. In addition, presence of Omp30 and absence of Omp6 seemed to be associated with H. pylori strains causing duodenal ulcer. PMID- 17081073 TI - Changes of hippocampal protein levels during postnatal brain development in the rat. AB - Information on postnatal brain protein expression is very limited, and we therefore compared hippocampal protein levels in rat hippocampus at different developmental time points using two-dimensional gel electrophoresis followed by mass spectrometrical protein identification and specific software for quantification. Proteins from several cascades as e.g., antioxidant, metabolic, cytoskeleton, proteasomal, and chaperone pathways were developmentally regulated, which is relevant for design and interpretation of protein chemical studies in the mammalian brain. PMID- 17081074 TI - Dysregulation of retinoid transporters expression in body fluids of schizophrenia patients. AB - This study aims to find the biomarkers or associated proteins in body fluids of schizophrenia patients so that we can further understand the etiology of schizophrenia. We applied proteomic technologies combining two-dimensional electrophoresis with Coomassie blue staining and mass spectrometry and identified a procedure for the clinical screening of disease-influenced body fluid proteins in two sets of samples, plasma from 19 schizophrenia patients and cerebrospinal fluid (CSF) from 35 drug-treated schizophrenic patients and 36 healthy controls. The expression of transthyretin (TTR) tetramer increased significantly in plasma of schizophrenic patients after a valid 2 months in-hospital antipsychotic treatment. Conversely, the expression of the TTR tetramer and apolipoprotein E (ApoE) was down-regulated by up to 1.68 and 3.62 times, respectively, in the CSF of schizophrenia patients compared to that of normal controls, which has not been reported previously. Considering that the TTR tetramer and ApoE are both retinoid transporters, retinoid dysfunction might be involved in the pathology of schizophrenia. PMID- 17081075 TI - Comorbid schizophrenia and substance abuse. AB - At some point during their lives, many patients with schizophrenia abuse substances. The co-occurrence of substance use leads to poorer long- and short term outcomes in schizophrenia and complicates the treatment of both conditions. The primary substances of abuse among schizophrenia patients are alcohol, cannabis, cocaine, and nicotine. This presentation describes the prevalence, outcomes, and basis for this comorbidity. A brief discussion about the neurobiology of schizophrenia explains how schizophrenia may create a biologic predisposition to substance abuse by altering the brain reward system. The efficacy of possible treatments for comorbid schizophrenia and substance abuse are weighed, including typical and atypical antipsychotics and psychosocial interventions, and a list of possible adjunctive agents is provided. PMID- 17081076 TI - Neuroprotection in schizophrenia. AB - Longitudinal and structural neuroimaging studies show that patients with schizophrenia that converted to psychosis were found to have progressive gray matter loss in the cortex. Gray matter loss was also associated with functional decline. While the underlying mechanisms of gray matter loss remain uncertain, evidence of improved outcomes suggests neuroprotection, the maintenance of the functional integrity of the brain in response to neurobiological stress, in schizophrenia is possible. In order to protect against gray matter loss and slow functional decline following the onset of psychosis, new data suggests that an appropriate antipsychotic chosen at first episode can modify the rate of structural deterioration, which can lead to improved outcome. PMID- 17081077 TI - Update on bipolar disorder and substance abuse: recent findings and treatment strategies. AB - Between 40% and 70% of people with bipolar disorder have a history of substance use disorder. A current or past comorbid substance use disorder may lead to worse outcomes for bipolar disorder, including more symptoms, more suicide attempts, longer episodes, and lower quality of life. Unfortunately, few treatments have been studied in patients with both illnesses, and large controlled trials are needed. Evidence from small studies suggests that some treatments proven for bipolar disorder (e.g., divalproex, lithium, quetiapine, lamotrigine, and psychotherapy) may decrease substance abuse or dependence. Both the bipolar disorder and the substance use disorder should be considered when determining the best management strategy. Once treatment has begun, clinicians should ensure that medication and psychotherapy are administered appropriately and that treatment is modified when there is inadequate response. PMID- 17081078 TI - The neurobiology of cognition in schizophrenia. AB - Cognitive dysfunction in schizophrenia differs from cognitive dysfunction in neurodegenerative illnesses because it is associated with neuronal dysfunction and not neurodegeneration. Pharmacologically, potential targets for developing treatments may differ from cognition in dementing disorders. Several putative molecular targets for treating cognition in schizophrenia show promise, such as treatments that act on the D(1) receptor of the dopamine system; the 5HT(1A), 5HT(2A), and 5HT(6), receptors of the serotonin system; and ampakines, Glycine/D cycloserine, D-serine, and mGluR 2/3 agonists of the glutamatergic system. Other receptors associated with improvement in cognition include nicotinic and muscarinic receptors, and the alphalpha2 subunit receptor of the brain GABA system. Domain treatment of schizophrenia is a new method of treating schizophrenia that involves treating a single domain of dysfunction at a time. PMID- 17081079 TI - Kidney disease and cardiovascular risk. AB - Kidney failure and chronic kidney disease (CKD) are associated with accelerated cardiovascular disease, apparently because of a high burden of traditional vascular risk factors and possibly nontraditional risk factors such as inflammation, chronic volume overload, and abnormal calcium-phosphate metabolism. Although the burden of cardiovascular disease in CKD patients is well documented, potentially beneficial therapies appear to be underused in mild to moderate CKD and are relatively understudied in those with kidney failure. This review describes the epidemiology and pathophysiology of cardiovascular disease in CKD. We also discuss the clinical and public health implications of current knowledge and outline opportunities for further research. PMID- 17081080 TI - Novel role of kallistatin in protection against myocardial ischemia-reperfusion injury by preventing apoptosis and inflammation. AB - Kallistatin is a serine proteinase inhibitor that has been shown to reduce joint swelling and to inhibit inflammation in a rat model of arthritis. In this study, we investigated the effect and mechanisms of kallistatin on cardiac function after myocardial ischemia-reperfusion (I/R) injury. The human kallistatin gene in an adenoviral vector was delivered locally into rat heart 4 days before 30-min ischemia followed by 24-hr reperfusion. Kallistatin gene transfer significantly reduced myocardial infarct size and left ventricle end-diastolic pressure and improved cardiac contractility. Kallistatin significantly reduced I/R-induced cardiomyocyte apoptosis as identified by TUNEL and Hoechst staining, DNA laddering, cell viability, and caspase-3 activity in ischemic myocardium and in primary cultured cardiomyocytes. Kallistatin also reduced intramyocardial monocyte/macrophage and neutrophil accumulation in conjunction with decreased expression of monocyte chemoattractant protein-1, tumor necrosis factor-alpha, and intercellular adhesion molecule-1. Kallistatin delivery promoted cardiac endothelial nitric oxide synthase activation and increased nitric oxide (NO) formation, but inhibited NADH oxidase activity, p22phox expression, and superoxide production. Moreover, kallistatin reduced the phosphorylation of apoptosis signal-regulating kinase-1 and mitogen-activated protein kinases (MAPKs), but increased Akt and glycogen synthase kinase-3beta phosphorylation. The effects of kallistatin on cardiac function, oxidative stress, and these signal transduction events were all blocked by Nomega-nitro-L-argi-nine methyl ester. These results indicate a novel role of kallistatin in cardiac protection after I/R injury through increased NO formation and Akt-glycogen synthase kinase 3beta signaling and suppression of oxidative stress and MAPK activation. PMID- 17081081 TI - Management of recalled pacemakers and defibrillators. PMID- 17081082 TI - Patient subpopulations with elevated cardiovascular risk pose unique treatment challenges. PMID- 17081084 TI - Effective treatment of hypertension by AT(1) receptor antagonism: the past and future of telmisartan. AB - Lowering blood pressure is the most effective treatment method to ensure a reduction in the total risk for cardiovascular morbidity and mortality. The renin angiotensin system plays an important role in volume homeostasis and blood pressure regulation and is a target for several groups of pharmaceutical agents. Angiotensin II receptor blockers represent the newest class of antihypertensive compounds. They prevent the binding of angiotensin II to the subtype 1 receptor (AT(1)), which is believed to mediate most of the physiological actions relevant to the regulation of blood pressure. Telmisartan, a widely used AT(1) receptor antagonist, is a highly selective compound with high potency, a long duration of action and a tolerability profile similar to placebo. Numerous randomized clinical trials and community-based studies have demonstrated that oral telmisartan and combinations of telmisartan with hydrochlorothiazide are at least as effective in lowering blood pressure as all other hypertensive medications. This has been demonstrated in different populations of adult patients with mild to-moderate essential hypertension, including patients with coexisting Type 2 diabetes, metabolic syndrome or renal impairment. Several large-scale, long-term, clinical endpoint studies are in progress to assess the beneficial effects of telmisartan on hypertension-related end-organ damage in patients at high risk of renal, cardiac and vascular damage whose blood pressure is well controlled. The most recent data from clinical trials and latest research regarding telmisartan will be reviewed in this article. PMID- 17081085 TI - Use of the ACE inhibitor zofenopril in the treatment of ischemic heart disease. AB - Zofenopril, an inhibitor of the angiotensin-converting enzyme (ACE), has recently been widely introduced into the pharmaceutical market. Its clinical safety and efficacy has been demonstrated in patients with hypertension and in patients with acute myocardial infarction (AMI). The Survival of Myocardial Infarction Long term Evaluation (SMILE) project provided valuable information regarding the safety of early onset ACE inhibition with zofenopril after AMI and a greater perception of the early and late benefits. The SMILE-I study demonstrated that most benefits of ACE inhibition may be obtained early after AMI and persist after discontinuation of treatment. The SMILE-II study demonstrated that early zofenopril treatment (initiated <12 h) is safe and associated with a low rate of severe hypotension in thrombolyzed patients with acute myocardial infarction when administered in accordance with an adequate dose-titration scheme. Many other studies of clinical ACE-inhibitors (ACEIs) over the last 30 years have provided us with information in order to understand the effects of ACEIs and have demonstrated that patients benefit from ACEI treatment at different stages of the pathophysiological continuum of cardiovascular diseases. The current guidelines recommend that ACEIs should be used for routine secondary prevention in all patients with coronary artery disease and should be considered for all other patients with coronary or other vascular disease unless contraindicated. PMID- 17081086 TI - The use of TP10, soluble complement receptor 1, in cardiopulmonary bypass. AB - Cardiopulmonary bypass (CPB) for cardiac surgery or lung transplantation initiates a systemic inflammatory response characterized by increased vascular permeability, generalized edema, abnormal lung function and oxygenation and impaired ventricular function. This post-CPB syndrome significantly contributes to postoperative morbidity and mortality. Activation of complement during CPB is a key component that initiates and augments this process. TP10, soluble complement receptor 1, is a novel complement inhibitor that is a potent inhibitor of C3 and C5 convertases, blocking activation of the complement cascade at the nexus of all three complement pathways. Recent controlled trials in humans have demonstrated that TP10 effectively inhibits complement activation during CPB. In high-risk adult patients, TP10 decreases the incidence of mortality and myocardial infarction in males but not in females following cardiac surgery. TP10 is also well tolerated and protects vascular function in infants undergoing CPB. In addition, TP10 leads to early extubation in adult lung transplant recipients. TP10 is currently positioned for clinical development in a male-only indication of cardiac surgery on CPB. PMID- 17081087 TI - Renal safety of iodixanol. AB - Iodinated contrast is a mainstay for diagnostic and interventional procedures performed by cardiologists, radiologists and other specialists. With the emergence of computed tomographic techniques for the evaluation of cardiac disease, malignancies, trauma and a variety of other internal disorders, the use of iodinated contrast is expected to increase dramatically over the next few years. There has been considerable refinement over the past decades from ionic high-osmolar, to nonionic low-osmolar and finally to nonionic iso-osmolar contrast. Iodixanol is the only nonionic iso-osmolar contrast approved for intravascular use. This contrast agent has the lowest rates of systemic and renal adverse events. Clinical trials have demonstrated the lowest rates of contrast induced nephropathy among all currently available forms of iodinated contrast. Specifically, iodixanol has been associated with a 71% relative risk reduction for contrast-induced nephropathy compared with low-osmolar agents in head-to-head randomized trials. This article reviews the structure, pharmacology and outcomes associated with iodixanol. PMID- 17081088 TI - Pacing in adults with congenital heart disease. AB - Adults with congenital heart disease constitute one of the fastest growing populations in cardiology. Pacing is an integral part of their therapy and may reduce their morbidity and mortality significantly. The current generation of pacemakers is more sophisticated and complex, and they are being utilized for indications other than conduction abnormalities, such as termination of tachycardia and improvement of heart failure. The complex anatomy and history of multiple previous surgeries in adults with congenital heart disease, however, pose many limitations and technical challenges related to the placement of a pacemaker. Unique and innovative approaches to endocardial lead placement and improved epicardial leads is making pacemaker implantation more feasible in these patients. PMID- 17081089 TI - Percutaneous closure of ventricular septal defects. AB - Isolated ventricular septal defect is the most common form of congenital heart disease. Surgery has been performed for many years and is considered the gold standard for the treatment of ventricular septal defects. However, it is associated with morbidity and mortality. Less invasive techniques have been developed and currently percutaneous closure of muscular and perimembranous ventricular septal defects has to be considered a possible alternative to the standard surgical approach. PMID- 17081090 TI - Minimally invasive approach to the child with palpitations. AB - Palpitations are a common reason for referral to a pediatric cardiologist. Although generally benign, palpitations are a significant cause for concern in the individual and their family. Similarly, palpitations may be the initial presentation of significant heart disease, resulting in heightened concern in the referring physician. Although emphasis is usually placed on excluding arrhythmia as the cause for palpitations, there are a variety of noncardiac causes for palpitations. The patient history and physical examination are the key components of the evaluation and guide subsequent investigations. In many cases, an immediate diagnosis cannot be made and additional testing may be required; this often includes further monitoring for episodes, cardiac imaging and ambulatory monitoring. Current technologies for ambulatory monitoring during symptoms include Holter monitoring and a variety of patient-activated event recorders, including implantable loop recorders. Each presents its own unique advantages and disadvantages to aid diagnosis in the management of a child with palpitations. The primary focus for the clinician is to determine whether the etiology is benign in nature or whether there is underlying heart disease that may carry a more serious prognosis. PMID- 17081091 TI - Immune response to allograft implantation in children with congenital heart defects. AB - Cryopreserved valved allografts are frequently used in the repair of congenital heart defects in children. Although the longevity of these grafts is generally good in most patients, there continue to be ongoing problems with allograft dysfunction and subsequent failure, particularly in infants and young children. The aim of this review is to discuss the immunogenicity of cryopreserved allograft tissue and measures that may minimize the deleterious effect of the immune system on allograft function and durability. PMID- 17081092 TI - Identification of atherosclerosis-modifying genes: pathogenic insights and therapeutic potential. AB - Atherosclerosis is a common, complex trait, with genetic variation in many genes and the environment contributing to risk for this condition, which has multiple and highly variable phenotypic manifestations. Progress has been made in the identification of atherosclerosis-modifying genes in both human studies and through the use of animal models. In the future, it may be possible to administer a genetic test for variation in a handful of the most common atherosclerosis modifier genes, and thus predict if a patient is likely to develop atherosclerosis and ischemic heart disease. These patients could then be treated aggressively to lower their low-density lipoprotein cholesterol levels and other risk factors, including counseling to make adjustments in their lifestyle. This review will summarize the methods and results thus far in the identification of atherosclerosis-modifier genes. PMID- 17081093 TI - Transcatheter mapping and ablation of arrhythmias in the coronary sinus. AB - The coronary sinus (CS) is a complex structure of particular interest to cardiac electrophysiologists. It is exploited as an anatomical reference, a site to record left-sided atrial and ventricular signals and for cardiac resynchronization therapy. Perhaps less appreciated, it may itself serve as a substrate for arrhythmias. It is now increasingly recognized that arrhythmias may be targeted by transcatheter ablation within the CS. This review summarizes pertinent anatomic considerations, explores the relationship between the CS and various arrhythmia substrates, elaborates on current indications for intra CS ablation and addresses efficacy and safety concerns associated with transcatheter ablation. PMID- 17081094 TI - Contegra conduit: current outcomes and concerns. AB - A variety of congenital cardiac anomalies with severe right ventricular outflow tract (RVOT) obstruction or RVOT interruption require surgical reconstruction from the infundibulum up to the pulmonary artery bifurcation or even into the branches of the pulmonary arteries. Ideally, the conduit or valve required for such reconstruction has to be formed of autologous tissue that grows, resists infection, lasts for the life span of the patient and is readily available in all sizes. Such conduits, however, are not available and although several alternatives have been used, none of which are without potential drawbacks. Contegra valved bovine internal jugular vein conduit (Medtronic Inc., MN, USA) has recently emerged as a promising option for pediatric RVOT reconstruction and has been advocated for its 'off-the-shelf' availability in sizes ranging from 12 to 22 mm, surgical pliability and encouraging short- and mid-term success in experimental animal, as well as clinical studies. This review focuses on the current outcomes of Contegra conduit and highlights some of the major concerns related to the use of this conduit and strategies to tackle these concerns. PMID- 17081095 TI - Perioperative cardiac risk: pathophysiology, assessment and management. AB - Cardiac complications are the leading cause of perioperative morbidity and mortality following noncardiac surgery. The annual cost of perioperative cardiovascular events exceeds 20 billion US dollars. A strategic preoperative evaluation holds the potential to reduce perioperative cardiac events and healthcare costs; however, our current understanding of the pathophysiological basis of postoperative acute coronary syndromes is limited. Although significant advances continue to facilitate early and reliable noninvasive detection of high risk coronary anatomy, the most appropriate interventions remain unclear. Pharmacotherapy, revascularization, safer anesthesia and early detection of perioperative heart failure may all reduce perioperative morbidity and mortality, although the evidence base is incomplete and controversial. A close working relationship between the primary care physician, cardiologist, surgeon and anesthesiologist will facilitate rational, tailored and optimized management decisions that constitute our best opportunity to reduce perioperative cardiovascular risk. PMID- 17081096 TI - Targeting angiogenesis versus myogenesis with cardiac cell therapy. AB - Considerable hope has been vested in cell therapy strategies designed to augment the endogenous neovascularization response to obstructive coronary artery disease, and to replace cardiomyocyte loss caused by myocardial infarction. Conceptually, the relative importance of targeting angiogenesis versus myogenesis in this scheme will vary depending on the clinical context (the predominance of ischemia versus ventricular dysfunction and scarring). Although the evidence so far is encouraging, whether these processes can be effectively targeted in a selective fashion with cell therapy is still unclear. Intriguingly, data are now emerging suggesting that the beneficial effects of cardiac cell therapies in a variety of clinical settings may be accounted for by a greater interaction of angiogenesis, myocardial salvage and myogenesis than heretofore appreciated, and through mechanisms that may include both cellular and paracrine effects. Greater understanding of these mechanisms should accelerate the development of effective cell therapies for the growing number of patients with advanced, and in many cases 'no-option', cardiovascular disease. Possible clinical targets for angiogenic and myogenic cardiac cell therapy, the scientific rationale for this therapeutic approach and future directions in this field are discussed here. PMID- 17081097 TI - My health: whose responsibility? Low-dose aspirin and older people. AB - The benefit of aspirin as a prophylactic after a thrombotic event was first observed 30 years ago. Its use after coronary or cerebral thrombosis, and in patients judged to be at increased risk of a thrombotic event, is now virtually mandatory, unless there are signs of intolerance. The present policy in the UK for cardiovascular protection by low-dose aspirin is dependent upon the identification of people at high vascular risk. The policy has had only very limited success, partly owing to the fact that only a relatively small proportion of people with levels of vascular risk factors that would justify aspirin prophylaxis are identified. In fact, it has been demonstrated that the application of accepted guidelines for aspirin prophylaxis to risk factor data in representative UK population samples gives a cost-effective evidence-base for a reasonable extension of prophylaxis to all people aged over approximately 50 years. It is possible that reductions in both dementia and cancer incidence could also follow the wider use of low-dose aspirin but further research on these outcomes is urgently required. The evidence on possible benefits and harm from low-dose aspirin should therefore be publicized widely, and everything possible should be done to stimulate discussion involving the general public. In the end, however, the preservation of health is one's own responsibility and, therefore, people should generally be encouraged to evaluate the evidence on health promotion measures, including low-dose aspirin, and take responsibility for their own health. PMID- 17081100 TI - Paper of the Year 2005: Award to Vanessa Ferreira Merino. PMID- 17081101 TI - Two-site substrate recognition model for the Keap1-Nrf2 system: a hinge and latch mechanism. AB - Cells are equipped with a number of transcriptional factors that safeguard against various environmental insults. Proteasomal protein degradation plays an important role in the Keap1-Nrf2 cytoprotection system, with molecular machinery similar to that for other environmental defense systems such as inflammatory and hypoxic responses. While Nrf2 protein stabilization is known to be redox sensitive, the transcription factors NF-kappaB and HIF-1alpha for inflammatory and hypoxic responses, respectively, are also influenced by the cellular redox conditions. In this review we present the recently proposed two-site substrate recognition model of the Keap1-Nrf2 system, which regulates the cellular responses against oxidative and xenobiotic stresses. The implications of two destructive motifs in Nrf2, the ETGE and DLG motifs, which appear to function as a hinge and latch attenuating Keap1 activity in different redox states, are discussed. PMID- 17081102 TI - Hypoxia and lipid signaling. AB - Sufficient oxygen supply is crucial for the development and physiology of mammalian cells and tissues. When simple diffusion of oxygen becomes inadequate to provide the necessary flow of substrate, evolution has provided cells with tools to detect and respond to hypoxia by upregulating the expression of specific genes, which allows an adaptation to hypoxia-induced stress conditions. The modulation of cell signaling by hypoxia is an emerging area of research that provides insight into the orchestration of cell adaptation to a changing environment. Cell signaling and adaptation processes are often accompanied by rapid and/or chronic remodeling of membrane lipids by activated lipases. This review highlights the bi-directional relation between hypoxia and lipid signaling mechanisms. PMID- 17081103 TI - Glutathione peroxidases and redox-regulated transcription factors. AB - Analysis of the selenoproteome identified five glutathione peroxidases (GPxs) in mammals: cytosolic GPx (cGPx, GPx1), phospholipid hydroperoxide GPx (PHGPX, GPx4), plasma GPx (pGPX, GPx3), gastrointestinal GPx (GI-GPx, GPx2) and, in humans, GPx6, which is restricted to the olfactory system. GPxs reduce hydroperoxides to the corresponding alcohols by means of glutathione (GSH). They have long been considered to only act as antioxidant enzymes. Increasing evidence, however, suggests that nature has not created redundant GPxs just to detoxify hydroperoxides. cGPx clearly acts as an antioxidant, as convincingly demonstrated in GPx1-knockout mice. PHGPx specifically interferes with NF-kappaB activation by interleukin-1, reduces leukotriene and prostanoid biosynthesis, prevents COX-2 expression, and is indispensable for sperm maturation and embryogenesis. GI-GPx, which is not exclusively expressed in the gastrointestinal system, is upregulated in colon and skin cancers and in certain cultured cancer cells. GI-GPx is a target for Nrf2, and thus is part of the adaptive response by itself, while PHGPx might prevent cancer by interfering with inflammatory pathways. In conclusion, cGPx, PHGPx and GI-GPx have distinct roles, particularly in cellular defence mechanisms. Redox sensing and redox regulation of metabolic events have become attractive paradigms to unravel the specific and in part still enigmatic roles of GPxs. PMID- 17081104 TI - Redox regulation of the hypoxia-inducible factor. AB - Reactive oxygen species (ROS) have long been considered only as cyto- and genotoxic. However, there is now compelling evidence that ROS also act as second messengers in response to various stimuli, such as growth factors, hormones and cytokines. The hypoxia-inducible transcription factor (HIF) is a master regulator of oxygen-sensitive gene expression. More recently, HIF has also been shown to respond to non-hypoxic stimuli. Interestingly, recent reports indicate that ROS regulate HIF stability and transcriptional activity in well-oxygenated cells, as well as under hypoxic conditions. Consequently, ROS appear to be key players in regulating HIF-dependent pathways under both normal and pathological circumstances. This review summarizes the current understanding of the role of ROS in the regulation of the mammalian HIF system. PMID- 17081105 TI - The L-arginine nitric oxide pathway: avenue for a multiple-level approach to assess vascular function. AB - Endothelial dysfunction is an early stage of atherosclerosis and has been attributed to impaired nitric oxide (NO) bioactivity and enhanced formation of oxygen-derived free radicals. Given that endothelial dysfunction is at least in part reversible, the assessment of altered NO availability is of important diagnostic and prognostic significance. Identification of such alterations may help to target asymptomatic individuals who are at risk for cardiovascular diseases and would likely benefit from preventive measures. Focusing on a single signaling pathway, we present here a multi-level approach for the early diagnosis of cardiovascular diseases by assessing molecular, biochemical, structural, and functional changes in the vascular wall. PMID- 17081106 TI - Protein oxidation and proteolysis. AB - One of the hallmarks of chronic or severe oxidative stress is the accumulation of oxidized proteins, which tend to form high-molecular-weight aggregates. The major proteolytic system responsible for the removal of oxidized cytosolic and nuclear proteins is the proteasome. This complicated proteolytic system contains a core proteasomal form (20S proteasome) and several regulators. All of these components are affected by oxidative stress to various degrees. The ATP-stimulated 26S proteasome is sensitive to oxidative stress, whereas the 20S form seems to be more resistant. The nuclear proteasome selectively degrades oxidatively damaged histones in the nuclei of mammalian cells, where it is activated and regulated by automodified PARP-1 after oxidative challenge. In this brief review we highlight the proteolysis and its regulatory effects during oxidative stress. PMID- 17081107 TI - Mitochondrial signaling, TOR, and life span. AB - Growing evidence supports the concept that mitochondrial metabolism and reactive oxygen species (ROS) play a major role in aging and determination of an organism's life span. Cellular signaling pathways regulating mitochondrial activity, and hence the generation of ROS and retrograde signaling events originating in mitochondria, have recently moved into the spotlight in aging research. Involvement of the energy-sensing TOR pathway in both mitochondrial signaling and determination of life span has been shown in several studies. This brief review summarizes the recent progress on how mitochondrial signaling might contribute to the aging process with a particular emphasis on TOR signaling from invertebrates to humans. PMID- 17081108 TI - Pathogenetic interplay between osmotic and oxidative stress: the hepatic encephalopathy paradigm. AB - Hepatic encephalopathy (HE) defines a primary gliopathy associated with acute and chronic liver disease. Astrocyte swelling triggered by ammonia in synergism with different precipitating factors, including hyponatremia, tumor necrosis factor (TNF)-alpha, glutamate and ligands of the peripheral benzodiazepine receptor (PBR), is an early pathogenetic event in HE. On the other hand, reactive nitrogen and oxygen species (RNOS) including nitric oxide are considered to play a major role in HE. There is growing evidence that osmotic and oxidative stresses are closely interrelated. Astrocyte swelling produces RNOS and vice versa. Based on recent investigations, this review proposes a working model that integrates the pathogenetic action of osmotic and oxidative stresses in HE. Under participation of the N-methyl-D-aspartate (NMDA) receptor, Ca(2+), the PBR and organic osmolyte depletion, astrocyte swelling and RNOS production may constitute an autoamplificatory signaling loop that integrates at least some of the signals released by HE-precipitating factors. PMID- 17081109 TI - Regulation of redox-sensitive exofacial protein thiols in CHO cells. AB - Thiols affect a variety of cell functions, an effect known as redox regulation, largely attributed to modification of transcription factors and intracellular signaling mechanisms. Since exofacial protein thiols are more exposed to redox acting molecules used in cell culture and may represent sensors of the redox state of the environment, we investigated their susceptibility to redox regulation. Exofacial protein thiols were measured using cell-impermeable Ellman's reagent [5,5'-dithiobis(2-nitrobenzoic acid), DTNB]. For quantification, we also set up an ELISA assay based on the cell-impermeable biotinylated SH reagent, N-(biotinoyl)-N-(iodoacetyl) ethylendiamine (BIAM). Exposure of CHO cells to H(2)O(2) induces oxidation of surface thiols at concentrations not affecting intracellular GSH. Depletion (50%) of GSH decreases surface thiols by 88%. Surface thiols are also highly sensitive to thiol antioxidants, since exposure to 5 mM N-acetyl-L-cysteine (NAC) for 2 h augmented their expression without increasing GSH levels. Using BIAM labeling and two-dimensional gel electrophoresis, we show that this increase in surface thiols is due to the reduction of specific membrane proteins. Peptide mass fingerprinting by MALDI mass spectrometry allowed us to identify two of these proteins as Erp57 and vimentin. PMID- 17081110 TI - N-ethylmaleimide-sensitive factor: a redox sensor in exocytosis. AB - Vascular injury triggers endothelial exocytosis of granules, releasing pro inflammatory and pro-thrombotic mediators into the blood. Nitric oxide (NO) and reactive oxygen species (ROS) limit vascular inflammation and thrombosis by inhibiting endothelial exocytosis. NO decreases exocytosis by regulating the activity of the N-ethylmaleimide-sensitive factor (NSF), a central component of the exocytic machinery. NO nitrosylates specific cysteine residues of NSF, thereby inhibiting NSF disassembly of the soluble NSF attachment protein receptor (SNARE). NO also modulates exocytosis of other cells; for example, NO regulates platelet activation by inhibiting alpha-granule secretion from platelets. Other radicals besides NO can regulate exocytosis as well. For example, H(2)O(2) inhibits exocytosis by oxidizing NSF. Using site-directed mutagenesis, we have defined the critical cysteine residues of NSF, and found that one particular cysteine residue, C264, renders NSF sensitive to oxidative stress. Since radicals such as NO and H(2)O(2) inhibit NSF and decrease exocytosis, NSF may act as a redox sensor, modulating exocytosis in response to changes in oxidative stress. PMID- 17081111 TI - Aspects of the biological redox chemistry of cysteine: from simple redox responses to sophisticated signalling pathways. AB - The last decade has witnessed an increased interest in cysteine modifications such as sulfenic and sulfinic acids, thiyl radicals, sulfenyl-amides and thiosulfinates, which come together to enable redox sensing, activation, catalysis, switching and cellular signalling. While glutathionylation, sulfenyl amide formation and disulfide activation are examples of relatively simple redox responses, the sulfinic acid switch in peroxiredoxin enzymes is part of a complex signalling system that involves sulfenic and sulfinic acids and interacts with kinases and sulfiredoxin. Although the in vivo evaluation of sulfur species is still complicated by a lack of appropriate analytical techniques, research into biological sulfur species has gained considerable momentum and promises further excitement in the future. PMID- 17081112 TI - Singlet oxygen inactivates protein tyrosine phosphatase-1B by oxidation of the active site cysteine. AB - Singlet oxygen ((1)O(2)), an electronically excited form of molecular oxygen, is a mediator of biological effects of ultraviolet A radiation, stimulating signaling cascades in human cells. We demonstrate here that (1)O(2) generated by photosensitization or by thermodecomposition of 3,3'-(1,4 naphthylidene)dipropionate-1,4-endoperoxide inactivates isolated protein tyrosine phosphatases (PTPases). PTPase activities of PTP1B or CD45 were abolished by low concentrations of (1)O(2), but were largely restored by post-treatment with dithiothreitol. Electrospray ionization mass spectrometry analysis of tryptic digests of PTP1B exposed to (1)O(2) revealed oxidation of active-site Cys215 as the only cysteine residue oxidized. In summary, (1)O(2) may activate signaling cascades by interfering with phosphotyrosine dephosphorylation. PMID- 17081113 TI - Regulatory effects of the mitochondrial energetic status on mitochondrial p66Shc. AB - p66(Shc) promotes apoptosis and controls the intracellular redox balance. A fraction of p66(Shc) exists within mitochondria, where it oxidizes cytochrome c to form hydrogen peroxide, which in turn induces mitochondrial permeability and apoptosis. However, cells tolerate p66(Shc) expression and accumulate oxidative damage under normal conditions, implying that the p66(Shc) functions must be tightly regulated. Here we review available knowledge on the regulation of p66(Shc) transcription, protein stabilization and post-translational modifications. In addition, we report novel investigations into the role of the mitochondrial import machinery on p66(Shc) activation, which highlight the energetic status of mitochondria as a crucial determinant of p66(Shc) function. PMID- 17081114 TI - Air pollution-associated fly ash particles induce fibrotic mechanisms in primary fibroblasts. AB - Air pollution is associated with a variety of respiratory and cardiovascular disorders, including fibrosis. To understand the possible molecular mechanisms underlying this observation, we examined the effect of particulate matter on primary fibroblasts, the key regulators of the extracellular matrix. Fly ash collected in an experimental waste incinerator was used as model particles for fine and ultrafine pollution components. Brief treatment of fibroblasts isolated from adult male Wistar rat hearts with fly ash triggered the immediate formation of intracellular reactive oxygen species (ROS). Using phospho-specific antibodies we observed activation of p38 MAP kinase, p44/42 MAP kinase (ERK1/2) and p70(S6) kinase. Prolonged incubation with fly ash increased the expression of collagen 1 and TGF-beta1, but decreased mRNA levels of MMP9 and TNF-alpha. Cell proliferation was inhibited at high concentrations of fly ash. An increase in the level of advanced glycation endproduct (AGE) modification of various cellular proteins after long-term treatment of cultured fibroblasts with fly ash was observed. The results of our study demonstrate that direct activation of fibroblasts by combustion-derived particles is a mechanism that may contribute to the adverse health effects of particulate air pollution. PMID- 17081115 TI - Incinerator fly ash provokes alteration of redox equilibrium and liberation of arachidonic acid in vitro. AB - Numerous epidemiological studies have associated exposure to ambient particulate matter (PM) with pulmonary and cardiovascular health effects. Macrophages as a part of the primary pulmonary defence system play a crucial role by generating pro- and anti-inflammatory mediators. The aim of the present study was to examine the effect of incinerator fly ash (MAF02) as a model of environmental particulate matter on the formation of reactive oxygen species (ROS) and their ability to induce oxidative stress in RAW264.7 macrophages. Furthermore, the liberation of arachidonic acid (AA) was observed. The interaction of MAF02 with macrophages caused increased mobilisation of AA, accompanied by enhanced expression of cyclooxygenase-2 (COX-2). The MAF02-induced AA liberation was found to depend on an increased intracellular calcium concentration. In addition, MAF02-induced liberation of AA was selectively blocked by an ERK1/2 pathway-specific inhibitor, while inhibition of the p38 MAPK activity had no effect. Fly ash was also observed to induce an increase in cellular glutathione (GSH) content and antioxidative enzyme haem oxygenase-1 (HO-1). In correlation, experiments with dichlorofluorescein demonstrated increased formation of ROS upon treatment with fly ash. In summary, incinerator fly ash induces oxidative stress to a certain extent, resulting in the onset of important mechanisms related to inflammation. PMID- 17081116 TI - Unique neuronal functions of cathepsin L and cathepsin B in secretory vesicles: biosynthesis of peptides in neurotransmission and neurodegenerative disease. AB - Proteases are required for the production of peptide neurotransmitters and toxic peptides in neurodegenerative diseases. Unique roles of the cysteine proteases cathepsin L and cathepsin B in secretory vesicles for the production of biologically active peptides have been demonstrated in recent studies. Secretory vesicle cathepsin L participates in the proteolytic conversion of proenkephalin into the active enkephalin, an opioid peptide neurotransmitter that mediates pain relief. Moreover, recent findings provide evidence that cathepsin B in regulated secretory vesicles participates in the production of toxic beta-amyloid peptides that are known to accumulate extracellularly in Alzheimer's disease brains. The neurobiological functions of cathepsins L and B demonstrate that these secretory vesicle cysteine proteases produce biologically active peptides. These results demonstrate newly identified roles for cathepsins L and B in neurosecretory vesicles in the production of biologically active peptides. PMID- 17081117 TI - Two novel mitochondrial and chloroplastic targeting-peptide-degrading peptidasomes in A. thaliana, AtPreP1 and AtPreP2. AB - Two novel metalloendopeptidases in Arabidopsis thaliana, AtPreP1 and AtPreP2, are responsible for the degradation of targeting peptides in mitochondria and chloroplasts. Both AtPreP1 and AtPreP2 contain ambiguous targeting peptides and are dually targeted to both organelles. The proteases also have the capacity to degrade unstructured peptides of up to 65 amino acid residues, but not small proteins. The catalysis occurs in a huge catalytic chamber revealed by the crystal structure of AtPreP1 at 2.1 A. The enzymes show a preference for basic and small uncharged amino acids or serines at the cleavage sites. Despite similarities in cleavage specificities, cleavage-site recognition differs for both proteases and is context- and structure-dependent. The AtPreP1 and AtPreP2 genes are differentially expressed in Arabidopsis. PMID- 17081118 TI - Switch from actin alpha1 to alpha2 expression and upregulation of biomarkers for pressure overload and cardiac hypertrophy in taurine-deficient mouse heart. AB - Taurine is the most abundant free amino acid in heart muscle and protects against heart failure. In the present study, the consequences of hereditary taurine deficiency on cardiac gene expression were examined in 2- and 15-16-month-old taurine transporter knockout (taut(-/-)) mice using a mouse-specific DNA microarray. This oligonucleotide-based microarray contains probes for 251 genes with relevance for heart function. Of these, 163 probes exhibited a reproducible hybridization signal and were analyzed. alpha-Actin type 1 mRNA levels were 70% lower in the heart of young and older taut(-/-) mice compared to wild-type controls. Interestingly, the hearts of taut(-/-) mice showed a switch from alpha actin 1 to alpha-actin 2 expression, as confirmed by real-time PCR and Western blot analysis. In addition, mRNA levels of biomarkers for pressure overload and hypertension were upregulated in taut(-/-) hearts, i.e., atrial natriuretic factor (+848%), brain natriuretic peptide (+90%), cardiac ankyrin repeat protein (+118%), and procollagen 1a1, 1a2 and 3a1 (+40% at least). These results point to a stress situation in the heart of taut(-/-) mice under laboratory conditions, and it can be speculated that taut(-/-) hearts may be even more susceptible to failure in the wild when under exogenous stress. PMID- 17081119 TI - Human RBM28 protein is a specific nucleolar component of the spliceosomal snRNPs. AB - The biogenesis of spliceosomal small nuclear RNAs (snRNAs) involves organized translocations between the cytoplasm and certain nuclear domains, such as Cajal bodies and nucleoli. Here we identify human RBM28 protein as a novel snRNP component, based on affinity selection of U6 small nuclear ribonucleoprotein (snRNP). As shown by immunofluorescence, RBM28 is a nucleolar protein. Anti-RBM28 immunoprecipitation from HeLa cell lysates revealed that this protein specifically associates with U1, U2, U4, U5, and U6 snRNAs. Our data provide the first evidence that RBM28 is a common nucleolar component of the spliceosomal ribonucleoprotein complexes, possibly coordinating their transition through the nucleolus. PMID- 17081120 TI - The beta12-beta13 loop is a key regulatory element for the activity and properties of the catalytic domain of protein phosphatase 1 and 2B. AB - The molecular architectures of the catalytic core of protein phosphatase 1 (PP1) and protein phosphatase 2B (PP2B) are similar, and both contain a beta12-beta13 loop that consists of non-conserved residues. A truncation mutant containing the PP2B catalytic domain has previously been constructed in our laboratory, and designated CNAa. In this study, the PP1 catalytic subunit (PP1c) and CNAa, as well as mutants with the corresponding loops exchanged, were investigated using multiple substrates. Deletion of the beta12-beta13 loop from Y272 to A279 of PP1c or from Y311 to K318 of CNAa resulted in inactive proteins. Loop exchange generated chimeric mutants called PP1-CNAa-loop and CNAa-PP1-loop. The activities and kinetic parameters of the two chimeric mutants were altered in the direction of the enzyme from which its loop was derived. The activity of PP1c or CNAa-PP1 loop was similar whether preincubated with Mn(2+) or not, while CNAa and PP1-CNAa loop can acquire enhanced activation if preincubated with Mn(2+) for longer periods of time. Intrinsic fluorescence spectra revealed that the three dimensional structure was altered as a result of exchanging the loops of PP1c and CNAa. In conclusion, the beta12-beta13 loop is one of the key regulatory elements in the catalytic domain for the activity and properties of PP1c and CNAa. PMID- 17081121 TI - DNA-binding properties of the recombinant high-mobility-group-like AT-hook containing region from human BRG1 protein. AB - The hBRG1 protein, a central ATPase of the human switching/sucrose non-fermenting (SWI/SNF) remodeling complex, has a catalytic ATPase domain, an AT-hook motif and a bromodomain. Bromodomains, found in many chromatin-associated proteins, recognize N-acetyl-lysine in histones and other proteins. The AT-hook motif, first described in the high-mobility group of non-histone chromosomal proteins HMGA1/2, is a DNA-binding motif. The AT-hook binds to the AT-rich DNA sequences in the minor groove of B-DNA in a non-sequence specific manner. AT-hook motifs have been identified in many other DNA-binding proteins. In this study we cloned and purified a fragment of hBRG1 encompassing the AT-hook region and the bromodomain. Nuclear magnetic resonance (NMR) and circular dichroism (CD) analyses show that the recombinant domains are structured. The functionality of subdomains was checked by assessing their interactions with N-acetylated peptides from histones and with DNA. Isothermal titration calorimetric (ITC) analysis demonstrates that the primary micromolar interaction is through the AT-hook motif. The AT-hook region binds to linear DNA by unwinding it. These properties resemble the characteristics of the HMGA1/2 proteins and their interaction with DNA. PMID- 17081122 TI - Papaya glutamine cyclotransferase shows a singular five-fold beta-propeller architecture that suggests a novel reaction mechanism. AB - Cyclisation of N-terminal glutamine and/or glutamate to yield pyroglutamate is an essential posttranslational event affecting a plethora of bioactive peptides and proteins. It is directly linked with pathologies ranging from neurodegenerative diseases to inflammation and several types of cancers. The reaction is catalysed by ubiquitous glutaminyl cyclotransferases (QCs), which present two distinct prototypes. Mammalian QCs are zinc-dependent enzymes with an alpha/beta-hydrolase fold. Here we present the 1.6-A-resolution structure of the other prototype, the plant analogue from Carica papaya (PQC). The hatbox-shaped molecule consists of an unusual five-fold beta-propeller traversed by a central channel, a topology that has hitherto been described only for some sugar-binding proteins and an extracellular nucleotidase. The high resistance of the enzyme to denaturation and proteolytic degradation is explained by its architecture, which is uniquely stabilised by a series of tethering elements that confer rigidity. Strikingly, the N-terminus of PQC specifically interacts with residues around the entrance to the central channel of a symmetry-related molecule, suggesting that this location is the putative active site. Cyclisation would follow a novel general-acid/base working mechanism, pivoting around a strictly conserved glutamate. This study provides a lead structure not only for plant QC orthologues, but also for bacteria, including potential human pathogens causing diphtheria, plague and malaria. PMID- 17081123 TI - First identification of a phosphorylcholine-substituted protein from Caenorhabditis elegans: isolation and characterization of the aspartyl protease ASP-6. AB - Caenorhabditis elegans is a widely accepted model system for parasitic nematodes, drug screening and developmental studies. Similar to parasitic worms, C. elegans expresses glycosphingolipids and glycoproteins carrying, in part, phosphorylcholine (PCho) substitutions, which might play important roles in nematode development, fertility and, at least in the case of parasites, survival within the host. With the exception of a major secretory/excretory product from Acanthocheilonema viteae (ES-62), no protein carrying this epitope has been studied in detail yet. Here we report on the identification, characterization and localization of the aspartyl protease ASP-6 of C. elegans, which is excreted by the nematode in a PCho-substituted form. Within the worm, most prominent expression of the protein is observed in the intestine, while muscle and epithelial cells express asp-6 to a lesser extent. In animals harboring an ASP 6::GFP fusion protein, diffuse fluorescence throughout the body cavity of adult worms indicates that the chimeric protein is secreted. PMID- 17081124 TI - The human cathelicidin peptide LL-37 and truncated variants induce segregation of lipids and proteins in the plasma membrane of Candida albicans. AB - The human cathelicidin peptide LL-37 and several truncated variants differ in their capability to transmigrate over the plasma membrane of Candida albicans. We investigated whether retention at the cell perimeter or membrane transmigration affects their membrane-disrupting activities and candidacidal properties. Using fluorescein-labeled peptides, we demonstrate that LL-37 and its C-terminally truncated peptide LL-31 remain permanently associated with the perimeter of the cell. The N-terminally truncated peptide RK-31 initially accumulated at the cell boundary, but transmigrated into the cytoplasm within 30 min. The C-terminally truncated peptide LL-25 transmigrated instantaneously into the cytoplasm. The ultrastructural effects on the plasma membrane were studied by freeze-fracture electron microscopy combined with filipin cytochemistry. All peptides, whether they transmigrated over the plasma membrane or not, induced phase separation in the plasma membrane. All peptides induced leakage of cell components, including nucleotides and proteins. Proteins were identified by SDS-PAGE in combination with mass spectrometry, which revealed that predominantly proteins smaller than 50 kDa had leaked out of C. albicans. PMID- 17081125 TI - Specificity of human cathepsin S determined by processing of peptide substrates and MHC class II-associated invariant chain. AB - Cathepsin S (CatS) is a lysosomal cysteine protease of the papain family, the members of which possess relatively broad substrate specificities. It has distinct roles in major histocompatibility complex (MHC) class II-associated peptide loading and in antigen processing in both the MHC class I and class II pathways. It may therefore represent a target for interference with antigen presentation, which could be of value in the therapy of (auto)immune diseases. To obtain more detailed information on the specificity of CatS, we mapped its cleavage site preferences at subsites S3-S1' by in vitro processing of a peptide library. Only five amino acid residues at the substrate's P2 position allowed for cleavage by CatS under time-limited conditions. Preferences for groups of amino acid residues were also observed at positions P3, P1 and P1'. Based on these results, we developed highly CatS-sensitive peptides. After processing of MHC class II-associated invariant chain (Ii), a natural protein substrate of CatS, we identified CatS cleavage sites in Ii of which a majority matched the amino acid residue preference data obtained with peptides. These observed cleavage sites in Ii might be of relevance for its in vivo processing by CatS. PMID- 17081126 TI - Mast cell-dependent activation of pro matrix metalloprotease 2: A role for serglycin proteoglycan-dependent mast cell proteases. AB - The formation of active matrix metalloprotease-2 (MMP-2) requires the proteolytic processing of proMMP-2, a process that can occur through the formation of a ternary complex between proMMP-2, the tissue inhibitor of metalloprotease-2 and membrane type 1-MMP. However, other activation mechanisms have been suggested, and in this study we investigated whether mast cells (MCs) may play a role in the activation of proMMP-2. Murine peritoneal cells, a mixture of macrophages, lymphocytes and MCs, were cultured ex vivo. Addition of proMMP-2 to resting peritoneal cell cultures resulted in only slow conversion of proMMP-2 into the active enzyme. However, when MC degranulation was provoked using a calcium ionophore, proMMP-2 processing was markedly enhanced. When the peritoneal cell populations were depleted in MCs, proMMP-2 processing was abrogated, but was reconstituted when purified MCs were added to the depleted cultures. ProMMP-2 processing was sensitive to serine protease inhibitors, but not to inhibitors of other classes of proteases. Furthermore, proMMP-2 processing was completely abrogated in cells lacking serglycin, a proteoglycan that has previously been shown to mediate storage of a variety of MC serine proteases. Taken together, these results suggest a novel mode of proMMP-2 activation mediated by serglycin dependent MC serine proteases. PMID- 17081140 TI - Spinal cord injury triggers systemic autoimmunity: evidence for chronic B lymphocyte activation and lupus-like autoantibody synthesis. AB - Clinical and experimental data indicate that spinal cord injury (SCI) elicits pathological T-cell responses. Implicit in these data, but poorly understood, is that B lymphocytes (B cells) also contribute to the delayed pathophysiology of spinal trauma. Here, for the first time, we show that experimental spinal contusion injury elicits chronic systemic and intraspinal B cell activation with the emergence of a B cell-dependent organ-specific and systemic autoimmune response. Specifically, using sera from spinal cord injured mice, immunoblots reveal oligoclonal IgG reactivity against multiple CNS proteins. We also show SCI induced synthesis of autoantibodies that bind nuclear antigens including DNA and RNA. Elevated levels of anti-DNA antibodies are a distinguishing feature of systemic lupus erythematosus and, via their ability to cross-react with neuronal antigens, can cause neuropathology. We show a similar pathologic potential for the autoantibodies produced after SCI. Thus, mammalian SCI produces marked dysregulation of B cell function (i.e. autoimmunity) with pathological potential. PMID- 17081141 TI - Kinetics of glial glutamine efflux and the mechanism of neuronal uptake studied in vivo in mildly hyperammonemic rat brain. AB - Kinetics of glial glutamine (GLN) transport to the extracellular fluid (ECF) and the mechanism of GLN(ECF) transport into the neuron--crucial pathways in the glutamine-glutamate cycle--were studied in vivo in mildly hyperammonemic rat brain, by NMR and microdialysis to monitor intra- and extracellular GLN. The minimum rate of glial GLN efflux, determined from the rate of GLN(ECF) increase during perfusion of alpha-(methylamino)isobutyrate (MeAIB), which inhibits neuronal GLN(ECF) uptake by sodium-coupled amino-acid transporter (SAT), was 2.88 +/- 0.22 micromol/g/h at steady-state brain [GLN] of 8.5 +/- 0.8 micromol/g. Our previous study showed that the rate of glutamine synthesis under identical experimental conditions was 3.3 +/- 0.3 micromol/g/h. At steady-state glial [GLN], this is equal to its efflux rate to the ECF. Comparison of the two rates suggests that SAT mediates at least 87 +/- 8% (= 2.88/3.3 x 100%) of neuronal GLN(ECF) uptake. While MeAIB induced > 2-fold elevation of GLN(ECF), no sustained elevation was observed during perfusion of the selective inhibitor of LAT, 2 amino-bicyclo[1,1,2]heptane-2-carboxylic acid (BCH), or of d-threonine, a putative selective inhibitor of ASCT2-mediated GLN uptake. The results strongly suggest that SAT is the predominant mediator of neuronal GLN(ECF) uptake in adult rat brain in vivo. PMID- 17081142 TI - Electroconvulsive seizure-induced gene expression profile of the hippocampus dentate gyrus granule cell layer. AB - Electroconvulsive shock (ECS) is the most effective treatment for depression, but the mechanism underlying the therapeutic action of this treatment is still unknown. To better understand the molecular changes that may be necessary for the clinical effectiveness of ECS we have combined the technologies of gene expression profiling using cDNA microarrays with T7-based RNA amplification and laser microdissection to identify regulated genes in the dentate gyrus granule cell layer of the hippocampus. We have identified genes previously reported to be up-regulated following ECS, including brain-derived neurotrophic factor, neuropeptide Y, and thyrotrophin releasing hormone, as well as several novel genes. Notably, we have identified additional genes that are known to be involved in neuroprotection, such as growth arrest DNA damage inducible beta (Gadd45beta), and the excitatory amino acid transporter-1 (EAAC1/Slc1A1). In addition, via in situ hybridization we show that EAAC1 is specifically up-regulated in the dentate gyrus, but not in other hippocampal subfields. This study demonstrates the utility of microarray analysis of microdissected subregions of limbic brain regions and identifies novel ECS-regulated genes. PMID- 17081144 TI - Nursing and nursing education in Thailand: the past, the present, and the future. PMID- 17081145 TI - Factor structure of the Maslach Burnout Inventory among Finnish nursing staff. AB - The purpose of the study was to investigate the factor structure of the Maslach Burnout Inventory-Human Services Survey (MBI-HSS) among Finnish nursing staff. Although the factorial validity of the MBI-HSS has been studied across different countries, the factor structure has never been investigated among Finnish nursing staff. The data were gathered by postal questionnaires from nursing staff working in health-care organizations around Finland. The sample consisted of 627 nurses and nurse managers. The factor structure was studied by using exploratory and confirmatory factor analysis. The fit of the hypothesized three-factor model to the data was superior to the alternative one-factor and two-factor models. The three-factor structure (emotional exhaustion, depersonalization, reduced personal accomplishment) showed the best fit, while the internal consistencies of the subscales were satisfactory. The MBI-HSS is a highly suitable instrument for measuring burnout among nursing staff and it is very applicable to Finnish health care research. PMID- 17081146 TI - Study on the relationship between backpack use and back and neck pain among adolescents. AB - A descriptive correlation study was conducted in Hobbs, New Mexico, USA, to find the relationship between backpack use and back and neck pain among adolescent boys and girls. A higher percentage of girls than boys rated their pain as being moderate to extremely strong. The correlation between pain and backpack weight per body mass index among girls was positive and significant, but negative and non-significant among boys. The correlation between pain and backpack weight was positive and non-significant among both boys and girls. The correlation between pain and each of the variables of backpack weight per body weight, backpack weight per height, and backpack weight per body surface area was negative and non significant among boys, but positive and non-significant among girls. Gender based research studies with additional variables and safe backpack use education in schools are recommended. PMID- 17081147 TI - Pacifiers: a microbial reservoir. AB - The permanent contact between the nipple part of pacifiers and the oral microflora offers ideal conditions for the development of biofilms. This study assessed the microbial contamination on the surface of 25 used pacifier nipples provided by day-care centers. Nine were made of silicone and 16 were made of latex. The biofilm was quantified using direct staining and microscopic observations followed by scraping and microorganism counting. The presence of a biofilm was confirmed on 80% of the pacifier nipples studied. This biofilm was mature for 36% of them. Latex pacifier nipples were more contaminated than silicone ones. The two main genera isolated were Staphylococcus and Candida. Our results confirm that nipples can be seen as potential reservoirs of infections. However, pacifiers do have some advantages; in particular, the potential protection they afford against sudden infant death syndrome. Strict rules of hygiene and an efficient antibiofilm cleaning protocol should be established to answer the worries of parents concerning the safety of pacifiers. PMID- 17081148 TI - Cardiac pain or panic disorder? Managing uncertainty in the emergency department. AB - Emergency department admissions for chest pain might be related to a psychological problem, such as panic. Panic disorder adversely affects the person's lifestyle, general health, and psychological well-being. Panic disorder is difficult to recognize, so nursing assessment and management of these clients is important. This paper presents research findings from a New Zealand study that explored emergency nurses' differentiation of non-cardiac chest pain from panic disorder and raised significant issues in the nursing assessment and management of such clients. The data were gathered from focus group interviews and were analyzed thematically. Three themes, prioritizing time, managing uncertainty and ambiguity, and the life-threatening lens, were identified. The findings confirm that a panic disorder is not always diagnosed when biomedical assessment is used in isolation from a psychosocial assessment. Emergency nurses are pivotal in reversing the cycle of repeat presenters with non-cardiac chest pain. Recommendations for assessing and managing this complex condition are presented. PMID- 17081149 TI - A historical comparison of the development of specialist genetic nursing in the United Kingdom and Japan. AB - This paper reports the development of specialist genetic nursing in the UK and Japan. Between 1970 and 1990, significant technical advances in genetic testing occurred. These advances offered many families affected by genetic conditions more choices in diagnostic, predictive, and prenatal testing. It was recognized that genetic testing could significantly alter the life of the family and that supportive health services were required to accompany such testing. Nurses began to offer specialist services in this setting, but considerable professional development was required to ensure practitioners were competent. In the UK, the profession grew slowly over several decades. Concern to establish minimum standards of practice in this new professional setting culminated in the establishment of a set of competencies and a formal registration process. By comparison, in Japan, developments in genetic nursing have occurred over a shorter period of time, but minimum standards of competence already have been agreed upon. PMID- 17081150 TI - Possibilities for cost containment in intensive care. AB - An intensive care unit (ICU) is valuable but consumes a disproportionately high amount of health-care resources. Accordingly, cost containment has been deemed a mandatory task. A review of the literature from many countries was completed to determine the strategies for reducing the cost of care in the ICU. The results of this review show that cost reduction can be achieved by using a variety of the following strategies: (i) instituting a closed ICU, where all the patient care is directed by intensivists or full-time critical care trained physicians; (ii) the utilization of interdisciplinary approaches to the care of patients in the ICU; (iii) developing and implementing a program of television-guided remote intensivists; (iv) the use of an alerting and reminding system; and (v) increasing the number of intermediate care beds for patients who require only monitoring and intensive nursing. The conclusion reached is that many of these strategies provide evidence for hospital manager decisions regarding cost containment strategies for the delivery of health care in the ICU. PMID- 17081151 TI - Neurologic and histologic outcome after intraneural injections of lidocaine in canine sciatic nerves. AB - BACKGROUND: Inadvertent intraneural injection of local anesthetics may result in neurologic injury. We hypothesized that an intraneural injection may be associated with higher injection pressures and an increase in the risk of neurologic injury. METHODS: The study was conducted in accordance with the principles of laboratory animal care, and was approved by the Laboratory Animal Care and Use Committee. Fifteen dogs of mixed breed (16-21 kg) were studied. After general endotracheal anesthesia, the sciatic nerves (n= 30) were exposed bilaterally. Under direct vision, a 25-gauge, long-beveled needle (30 degrees) was placed either epineurally (n= 10) or intraneurally (n= 20), and 4 ml of preservative-free lidocaine 20 mg/ml was injected using an automated infusion pump (4 ml/min). Injection pressure data were acquired using an in-line manometer coupled to a computer via an analog-to-digital conversion board. After injection, the animals were awakened and subjected to serial neurologic examinations. One week later, the dogs were killed, the sciatic nerves excised and histologic examination was performed by pathologists blind to the purpose of the study. RESULTS: All perineural injections resulted in low pressures (< or = 5 psi). In contrast, eight of 20 intraneural injections resulted in high pressures (20-38 psi) at the beginning of the injection. Twelve intraneural injections, however, resulted in pressures of less than 12 psi. Neurologic function returned to baseline within 3 h after perineural injections and within 24 h after intraneural injections, when the measured injection pressures were less than 12 psi. Neurologic deficits persisted throughout the study period after all eight intraneural injections that resulted in high injection pressures. Histologic examination of the affected nerves revealed fascicular axonolysis and cellular infiltration. CONCLUSIONS: The data in our canine model of intraneural injection suggest that intraneural injections do not always lead to nerve injury. High injection pressures during intraneural injection may be indicative of intrafascicular injection and may predict the development of neurologic injury. PMID- 17081152 TI - Malignant hyperthermia and central core disease causative mutations in Swedish patients. AB - BACKGROUND: Malignant hyperthermia (MH) susceptibility is a pharmacogenetic disorder of intracellular calcium homeostasis. In susceptible individuals, halogenated anaesthetics and/or suxamethonium may trigger an MH reaction. The diagnosis of MH susceptibility is made by an in vitro contracture test of biopsied muscle strips. METHODS: In 27 MH susceptible (MHS) probands and four MH negative (MHN) probands, exons 17, 39, 40, 45 and 46 of the RYR1 gene were screened for MH causative mutations. In addition, in three patients with established central core disease (CCD), exons 17, 39, 40, 45 and 46 and exons 95, 100, 101 and 102 were screened for MH and CCD causative mutations. All screenings were performed by direct sequencing of the entire exons. RESULTS: MH causative mutations were found in five of the 27 MHS probands (19%). CCD causative mutations were found in two of three CCD patients in the C-terminal exons. None of the CCD patients showed a mutation in N-terminal exon 17 or in the central exons. CONCLUSIONS: In a Swedish population, screening of N-terminal exon 17 and the central exons for MH causative mutations in the RYR1 gene covers 19% of families. Thus, other mutations must also be responsible for MH susceptibility in Sweden. Although the number of CCD patients in this study was small, screening of the C-terminal exons for CCD causative mutations seems to be a promising tool in the process of making a diagnosis. PMID- 17081153 TI - A life course perspective on telomere length and social inequalities in aging. AB - Longitudinal studies will be needed to test the idea that social class in adult life, or in childhood, influences the rate of change in telomere length in peripheral blood samples. PMID- 17081154 TI - Short telomeres: cause or consequence of aging? AB - Questions about mechanisms, about the direction of causality, and about cellular heterogeneity complicate interpretation of claims associating short telomeres with adverse health outcomes. PMID- 17081155 TI - How old are you now? AB - Cherkas et al. provide a new, biological approach to a classic problem in anthropology--the estimation of age. PMID- 17081156 TI - Stress, social rank and leukocyte telomere length. AB - Blood leukocytes are a heterogeneous mixture of cell types whose telomere lengths differ greatly, reflecting variation in stem cell turnover and recruitment, expansion and replacement of more mature cell types as well as variable telomere loss and telomere repair. These differences in cell and telomere length dynamics, together with the evidence that telomere length is influenced strongly by genetic polymorphisms, greatly complicate the interpretation of claims that socio economic status modulates the rate of telomere attrition. PMID- 17081157 TI - Uncoupling of pathways that promote postmitotic life span and apoptosis from replicative immortality of Caenorhabditis elegans germ cells. AB - A dichotomy exists between germ and somatic cells in most organisms, such that somatic cell lineages proliferate for a single generation, whereas the germ cell lineage has the capacity to proliferate from one generation to the next, indefinitely. Several theories have been proposed to explain the unlimited replicative life span of germ cells, including the elimination of damaged germ cells by apoptosis or expression of high levels of gene products that prevent aging in somatic cells. These theories were tested in the nematode Caenorhabditis elegans by examining the consequences of eliminating either apoptosis or the daf 16, daf-18 or sir-2.1 genes that promote longevity of postmitotic somatic cells. However, germ cells of strains deficient for these activities displayed an unlimited proliferative capacity. Thus, C. elegans germ cells retain their youthful character via alternative pathways that prevent or eliminate damage that accumulates as a consequence of cell proliferation. PMID- 17081158 TI - Haematopoietic stem cells and endothelial progenitor cells in healthy men: effect of aging and training. AB - The number of hematopoietic stem cells (HSC) and endothelial progenitor cells (EPC) is thought to be a marker for neovascularization and vascular repair. Because physical inactivity and aging are risk factors for cardiovascular diseases, these factors may influence the numbers of HSCs and EPCs. Therefore, we examined baseline and exercise-induced levels of HSCs and EPCs in sedentary and trained young and older men. To study the role of aging in eight sedentary young (19-28 years) and eight sedentary older men (67-76 years), baseline and acute exercise-induced numbers of HSCs (CD34+-cells) and EPCs (CD34+/VEGFR-2+-cells) were quantified by fluorescence-activated cell sorter (FACS) analysis. To examine the effect of chronic training, eight age-matched trained young men (18-28 years) were compared with sedentary young men, whereas older men performed an 8-week endurance training. Older men showed significantly lower baseline and exercise induced levels of HSCs/EPCs than the young men (P < 0.05). In young and older men, acute exercise significantly increased HSCs (P < 0.01), but not EPCs. The absolute increase in numbers of HSCs was attenuated in older men (P = 0.03). Apart from the lower baseline numbers of EPCs after chronic training in older men, training status did not alter baseline or exercise-induced levels of HSCs/EPCs in young and older men. We concluded that advancing age results in lower circulating numbers of HSCs and EPCs and attenuates the acute exercise induced increase in HSCs. Interestingly, in young as well as in older men chronic endurance training does not affect baseline and exercise-induced numbers of HSCs and EPCs. PMID- 17081159 TI - Role of protein kinase C-dependent A-kinase anchoring proteins in lysophosphatidic acid-induced cAMP signaling in human diploid fibroblasts. AB - Previously, we reported that lysophosphatidic acid (LPA)-induced adenosine 3',5' cyclic monophosphate (cAMP) production by human diploid fibroblasts depends on the age of the fibroblasts. In this study, we examined the role of A-kinase anchoring proteins (AKAP) in the regulation of LPA-stimulated cAMP production in senescent fibroblasts. We found that levels of protein kinase C (PKC)-dependent AKAPs, such as Gravin and AKAP79, were elevated in senescent cells. Co immunoprecipitation experiments revealed that Gravin and AKAP79 do not associate with adenylyl cyclase type 2 (AC2) but bind to AC4/6, which interacts with calcium-dependent PKCs alpha/beta both in young and senescent fibroblasts. When the expression of Gravin and AKAP79 was blocked by small interference RNA transfection, the basal level of cAMP was greatly reduced and the cAMP status after LPA treatment was also reversed. Protein kinase A showed a similar pattern in terms of its basal activity and LPA-dependent modulation. These data suggest that Gravin and to a lesser extent, AKAP79, may play important roles in maintaining the basal AC activity and in coupling the AC systems to inhibitory signals such as Gialpha in young cells, and to stimulatory signals such as PKCs in senescent cells. This study also demonstrates that Gravin is especially important for the long-term activation of PKC by LPA in senescent cells. We conclude that LPA-dependent increased level of cAMP in senescent human diploid fibroblasts is associated with increases in Gravin levels resulting in its increased binding with and activation of calcium-dependent PKC alpha/beta and AC4/6. PMID- 17081160 TI - Lifespan extension in Caenorhabditis elegans by complete removal of food. AB - A partial reduction in food intake has been found to increase lifespan in many different organisms. We report here a new dietary restriction regimen in the nematode Caenorhabditis elegans, based on the standard agar plate lifespan assay, in which adult worms are maintained in the absence of a bacterial food source. These findings represent the first report in any organism of lifespan extension in response to prolonged starvation. Removal of bacterial food increases lifespan to a greater extent than partial reduction of food through a mechanism that is distinct from insulin/IGF-like signaling and the Sir2-family deacetylase, SIR 2.1. Removal of bacterial food also increases lifespan when initiated in postreproductive adults, suggesting that dietary restriction started during middle age can result in a substantial longevity benefit that is independent of reproduction. PMID- 17081161 TI - Review article: interactions between genotype and response to therapy in inflammatory bowel diseases. AB - BACKGROUND: More than half of the patients with inflammatory bowel diseases are candidates for immunosuppressive therapy. However, even the most effective drugs used in inflammatory bowel disease are only successful in about two-thirds of patients. Adverse events limit their use in a further substantial proportion of patients. Recent research has focussed on the possibility of predicting a drugs' efficacy and/or toxicity by identifying polymorphic variants in the genes encoding enzymes involved in metabolic pathways. AIM: To highlight recent advances and limitations in the field of pharmacogenetics in inflammatory bowel disease. RESULTS: Recent pharmacogenetic studies have mainly focussed on immunosuppressive agents including corticosteroids, azathioprine, methotrexate and infliximab. Several polymorphic genes encoding enzymes involved in the metabolism of these drugs have been identified including the inosine triphosphate pyrophosphatase in thiopurine therapy, the methylene tetrahydrofolate reductase in methotrexate therapy and polymorphisms in apoptosis genes in infliximab therapy. However, at the present time, genotyping for the variants of the thiopurine methyltransferase gene, an enzyme important for the metabolism of the thiopurine drugs, is the only useful test in clinical practice. CONCLUSIONS: Although the field of pharmacogenetics in inflammatory bowel disease is promising most new targets have so far failed to translate into clinical practice. Future pharmaceutical trials should include pharmacogenetic research to test appropriate candidate genes in a prospective manner. PMID- 17081162 TI - Meta-analysis: anti-viral therapy of hepatitis C virus-related liver disease in renal transplant patients. AB - BACKGROUND: The efficacy and safety of interferon-based therapy in renal transplant recipients with hepatitis C remains unclear, although a number of small clinical trials have been published addressing this issue. AIM: To evaluate efficacy and safety of antiviral therapy with interferon (interferon alone or interferon plus ribavirin) in renal transplant patients with hepatitis C by performing a systematic review of the literature with a meta-analysis of clinical trials. METHODS: The primary outcomes were sustained virological response (as a measure of efficacy) and/or drop-out rate (as a measure of tolerability). We used the random-effects model of DerSimonian and Laird, with heterogeneity and sensitivity analysis. RESULTS: We identified 12 clinical trials (102 unique patients); there was one controlled study. The summary estimate for sustained virological response and drop-out rate was 18.0% (95% CI 7.0-29.0%) and 35.0% (95% CI 20-50%), respectively. The most frequent side-effect requiring interruption of treatment was graft dysfunction (n = 28; 71.7%). Meta-regression analysis showed an inverse and significant association between reference year and drop-out logit rate (P = 0.012); an inverse link between sustained virological response logit rate and frequency of hepatitis C virus genotype 1 (P = 0.067) and cirrhosis (P = 0.08) was found, even if no statistical significance was reached. No publication bias was observed. CONCLUSIONS: Interferon-based therapy of hepatitis C has poor tolerance and safety after renal transplant. The optimal treatment of hepatitis C after renal transplant requires additional agents or alternative therapeutic approaches. PMID- 17081163 TI - Nitazoxanide in the treatment of viral gastroenteritis: a randomized double-blind placebo-controlled clinical trial. AB - BACKGROUND: Enteric viruses including noroviruses and rotavirus are leading causes of diarrhoeal disease and gastroenteritis worldwide, and there is no effective treatment. AIM: To evaluate nitazoxanide, a thiazolide anti-infective agent, in treating viral gastroenteritis in adults and adolescents. METHODS: 50 out-patients at least 12 years of age (mean 33.5 years) presenting with diarrhoea and stool-positive by enzyme-linked immunosorbent assay for norovirus, rotavirus or adenovirus were enrolled in a double-blind, placebo-controlled clinical trial. Patients were randomly assigned either nitazoxanide 500 mg or placebo twice daily for 3 days. The primary end point was time from first dose to resolution of symptoms. Analysis was modified intent-to-treat for 45 patients, excluding five patients with other identified enteropathogens at baseline. RESULTS: The median time from first dose to resolution of symptoms was 1.5 days (IQR: 0.5-2.5) for nitazoxanide-treated patients and 2.5 days (IQR: 1.5-4.5) for the placebo group. Significant reductions in time to resolution of symptoms were observed for all patients analysed (P < 0.0001) and for subsets of patients with rotavirus (P = 0.0052) and norovirus (P = 0.0295). The number of patients with adenovirus (n = 5) was too small to draw any conclusion. No significant adverse events were reported. CONCLUSIONS: Nitazoxanide may play an important role in managing viral gastroenteritis in adults. PMID- 17081164 TI - An open-label study of rabeprazole in patients with Zollinger-Ellison syndrome or idiopathic gastric acid hypersecretion. AB - BACKGROUND: Omeprazole and lansoprazole are both of proven efficacy in the treatment of Zollinger-Ellison syndrome and idiopathic gastric acid hypersecretion. Rabeprazole, which has a similar mechanism of action, has not previously been studied in these diseases. AIM: To determine the dose of rabeprazole that decreased basal acid output to safe levels in patients with Zollinger-Ellison syndrome or idiopathic gastric acid hypersecretion. METHODS: Patients with Zollinger-Ellison syndrome or idiopathic gastric acid hypersecretion were given rabeprazole 60 mg once daily for uncomplicated disease or 40 mg twice daily for complicated disease. Doses were titrated according to response and continued for 2 years. Efficacy was assessed primarily by measuring basal acid output. RESULTS: All patients had basal acid output before the next dose controlled to <10 mmol/h either at the starting dose or after minor dose titration. Control of acid output was maintained for 2 years. Consistent with this, most patients reported few gastrointestinal symptoms. Gastric biopsy showed no enterochromaffin-like cell dysplasia or neoplasia. CONCLUSIONS: Rabeprazole was an effective and well-tolerated treatment for Zollinger-Ellison syndrome or idiopathic gastric acid hypersecretion, which reliably reduced gastric acid output to safe levels. Although a dose of 60 mg once daily was appropriate for most patients in this study, doses may need adjustment according to individual response. PMID- 17081165 TI - Cost-utility analysis of different peg-interferon alpha-2b plus ribavirin treatment strategies as initial therapy for naive Chinese patients with chronic hepatitis C. AB - BACKGROUND: Different peg-interferon alpha-2b plus ribavirin treatment strategies are more effective in treating hepatitis C. However, no cost-effectiveness data have been published using the clinical data from the peg-interferon alpha-2b and ribavirin in the treatment of patients with hepatitis C in Taiwan. AIM: To estimate the cost-effectiveness of different treatments with peg-interferon alpha 2b plus ribavirin for the initial treatment of patients with different genotype chronic hepatitis C. METHODS: Individual patient level data from a randomized clinical trial with peg-interferon plus ribavirin were applied to a Markov model to project lifelong clinical outcomes. Economic estimates and quality of life were based on published data and Taiwan patient data. We used a societal perspective and applied a 3% annual discount rate. RESULTS: Compared with different combination therapy strategies, peg-interferon alpha-2b plus weight based dosing of ribavirin in all patients for 24 weeks is the most cost-effective treatment strategy. If the sustained virological response of peg-interferon plus ribavirin treatment for 48 weeks therapy in genotype 1 patients was higher than 67.8%, the best strategy of treating patients will be the peg-interferon plus weight-based dosing of ribavirin therapy for 48 weeks in genotype 1 patients and for 24 weeks in non-genotype 1 patients. CONCLUSIONS: Peg-interferon alpha-2b plus ribavirin combination for 24 weeks therapy in all genotype patients should reduce the incidence of liver complications, prolong life, improve quality of life and be cost-effective for the initial treatment of chronic hepatitis C. PMID- 17081166 TI - Percutaneous radiofrequency thermal ablation for hepatocellular carcinoma. AB - BACKGROUND: Radiofrequency thermal ablation is the first therapeutic option in percutaneous treatment of hepatocellular carcinoma but data on its long-term efficacy and safety are not conclusive. AIM: This study reports a prospective survey on radiofrequency thermal ablation in north-east Italy. METHODS: Data were collected on 401 patients with hepatocellular carcinoma (males 301, mean age: 68 years) treated by radiofrequency thermal ablation in 13 centres. Indication to treatment was: single nodule not eligible for surgery in 77% of patients, 2-3 nodes in 18% and multiple lesions in 5%. Mean size was 3 cm (1-8 cm). Treatment response was assessed at 1 month by spiral computerized tomography and then with ultrasound examination and new spiral computerized tomography. RESULTS: Complete response was obtained in 67% of patients and in 27% response was 75-99%. Complete response raised to 77% in lesions smaller than 3 cm. The morbidity rate was 34%; the mortality was 0.5%, seeding was observed in four patients. Ten patients presented an unexpected rapid disease progression. CONCLUSION: The above data show that by radiofrequency thermal ablation, complete response can be achieved only in about two-third of the cases, clearly less than expected, and that, beyond seeding, unexpected progression can be observed. PMID- 17081167 TI - Bilirubin and colorectal cancer. PMID- 17081169 TI - Excellent therapy for Helicobacter pylori infection. PMID- 17081171 TI - Protection of spermatogenesis in mice against gamma ray induced damage by Hippophae rhamnoides. AB - The radioprotective action of a preparation from Hippophae rhamnoides berries RH 3, already reported to render >80% survival against whole body 10 Gy gamma irradiation, was further investigated with respect to the testicular system. RH-3 was administered to mice 30 min before gamma irradiation (5 and 10 Gy) and histological parameters such as testis weight, sperm count, frequency of abnormal sperm, repopulation index, stem cell survival index and seminiferous tubular diameter were assessed on the 35th day. RH-3 administration partially countered radiation induced reduction in testis weight, sperm count, repopulation index and stem cell survival index (p < 0.01). The increase in the frequency of abnormal sperm (15.17 +/- 1.046%) caused by irradiation (5 Gy) was counteracted by pre irradiation treatment with RH-3, which significantly decreased the level of abnormal spermatozoa to 7.99 +/- 0.918% (p < 0.001), i.e. 52% abnormalities in comparison with 5 Gy irradiated group. RH-3 treatment alone did not elicit any toxic or adverse effect on the process of spermatogenesis. The present study suggests that RH-3 treatment protected spermatogenesis by enhancing the spermatogonial proliferation, enhancing the stem cell survival and reducing sperm abnormalities. The presence of polyphenolic flavonoids and tannins in the extract and the radical scavenging activity might be responsible for the radioprotective action of RH-3. PMID- 17081172 TI - Sperm morphology and aneuploidies: defects of supposed genetic origin. AB - As individuals with genetic sperm defects are intracytoplasmic sperm injection candidates, the study of the chromosomal constitution of their spermatozoa is of great interest. This study is a review of the current literature concerning fluorescence in situ hybridisation studies in spermatozoa with genetic sperm defect as 'round head', 'dysplasia of fibrous sheath' (DFS), 'primary ciliary dyskinesia' (PCD), the 'detached tail' and the 'absence of fibrous sheath'. Regarding sperm head defects, elevated XY disomy and diplodies were detected. Genetic defects affecting the sperm tail seemed to have a different correlation with chromosome meiotic segregation. Only chromosome 18, among the autosomes, was studied and the percentage of frequency of disomy was generally within the normal range. In the more frequently studied defect, DFS, the alterations in gonosome disomy and diploidy were recorded by different groups. Regarding PCD defects, elevated frequencies of disomy of sex chromosomes and diploidy were observed, whereas the absence of the fibrous sheath and the detached tail did not show any meiotic disturbance. The problem of genetic sperm defects should be seriously considered when these sperm are used for assisted reproduction, owing to the high risk of transmission of chromosomal imbalance and of mutations that could cause genetic sperm defects in offspring. PMID- 17081173 TI - Low testosterone levels and unimpaired melatonin secretion in young males with metabolic syndrome. AB - The interrelations between testosterone, insulin and melatonin levels in males with metabolic syndrome (MS) are still not clarified, especially in young age groups. The aim of the present study was to compare the testosterone serum levels in young men with MS to those in healthy controls, and to determine the possible changes in their melatonin rhythm, as well as the relation between melatonin, insulin and lipid profile. Fasting insulin and testosterone concentrations were measured in 10 healthy nonobese and 10 MS patients. Blood samples for melatonin, insulin and luteinizing hormone (LH) were collected at 19.00, 03.00 and 11.00 hours. A significant difference was found between the testosterone levels in controls and patients. Luteinizing hormone levels in both groups were similar, however, higher night LH levels in MS patients were observed. No changes in the melatonin concentrations of the two groups were found. In conclusion, total testosterone levels were significantly lower in young men with MS compared with healthy age-matched controls. Mild hypoandrogenia in hyperinsulinaemic patients was not related with changes in their melatonin levels. No alterations in the endogenous melatonin rhythm of the MS patients were found. PMID- 17081174 TI - Effect of smoking on seminal plasma ascorbic acid in infertile and fertile males. AB - This work aimed to assess the relationship of seminal ascorbic acid levels with smoking in infertile males. One hundred and seventy men were divided into four groups: nonobstructive azoospermia [NOA: smokers (n = 20), nonsmokers (n = 20)]; oligoasthenozoospermia [smokers (n = 30), nonsmokers (n = 20)]; asthenozoospermia [smokers (n = 20), nonsmokers (n = 20)] and normozoospermic fertile men [smokers (n = 20), nonsmokers (n = 20)]. The patients underwent medical history, clinical examination, conventional semen analysis and estimation of ascorbic acid in the seminal plasma calorimetrically. There was a significant decrease in the mean seminal plasma ascorbic acid levels in smokers versus nonsmokers in all groups (mean +/- SD; 6.03 +/- 2.18 versus 6.62 +/- 1.29, 7.81 +/- 1.98 versus 9.44 +/- 2.15, 8.09 +/- 1.98 versus 9.95 +/- 2.03, 11.32 +/- 2.15 versus 12.98 +/- 12.19 mg dl(-1) respectively). Fertile subjects, smokers or not, demonstrated significant higher seminal ascorbic acid levels than any infertile group. Seminal plasma ascorbic acid in smokers and nonsmokers was correlated significantly with sperm concentration (r = 0.59, 0.60, P < 0.001), sperm motility (r = 0.65, 0.55, P < 0.001) and negatively with sperm abnormal forms per cent (r = -0.53, -0.50, P < 0.001). Nonsignificant correlations were elicited with semen volume (r = 0.2, 0.09) or liquefaction time (r = 0.03, 0.06). It is concluded that seminal plasma ascorbic acid decreased significantly in smokers and infertile men versus nonsmokers and fertile men, and is significantly correlated with the main sperm parameters: count, motility and normal morphology. Also, cigarette smoking is associated with reduced semen main parameters that could worsen the male fertilizing potential, especially in borderline cases. PMID- 17081175 TI - Lessons learned from a sperm morphology quality control programme. AB - Although sperm morphology is the sperm parameter that is consistently correlated with in vitro and in vivo fertilisation, its value is often challenged due to discrepancies in reports. As sperm morphology is subjectively evaluated, the outcome of the report is entirely based on the experience and technical skill of the laboratory technician responsible for semen analyses. The study aimed to record the value of quarterly monitoring and refresher courses on morphology reading skills of technicians over a period of 40 months. Nineteen individuals from 12 different andrology laboratories were enrolled to a sperm morphology quality control programme after initial training sessions. Five of the 19 participants attended annual refresher courses. Two limits of error, namely +/- 0.5 SD and +/- 0.2 SD scores, were used to record technician deviation from the reference laboratory. Fifteen of the 19 individuals (78%) consistently reported sperm morphology readings that were within the +/- 0.5 SD limits of error. Furthermore, five of the participants, who attended annual refresher training courses, consistently reported results that were within the +/- 0.2 SD limits of error. For the first time, it has been illustrated that excellent sperm morphology reading skills can be achieved and maintained by initial training sessions, followed up with a continuous external quality control programme and annual refresher courses. PMID- 17081176 TI - A new technique to evaluate the ability of cryoprotectors to prevent premature acrosome reaction in human spermatozoa. AB - Acrosome reaction (AR) induced by low temperature has been used to evaluate sperm function; it correlates adequately with the fertilization percentages in vitro. In this study, the technique of AR induction by low temperature was used to evaluate the effect in the protection of the acrosome by cryopreservatives normally used in human semen cryopreservation. Donor sperm selected by use of the migration sedimentation technique was incubated in human tubal fluid medium, added to dimethyl sulphoxide 1 m, ethylene glycol 0.75 m, glycerol 1 m, incubated at 4 degrees C and 20 degrees C (as a control) for 18 h, and then for 3 h at 37 degrees C in a cell incubator. The AR was evaluated by triple stain in 100 viable spermatozoa. The effect of cryopreservatives on acrosome preservation in samples incubated for 18 h at 4 degrees C was as follows: 78% intact acrosome for glycerol, 77.8% intact acrosome for dimethyl sulphoxide and 96.2% intact acrosome for ethylene glycol (P < 0.0025 compared with glycerol and dimethyl-sulphoxide). The sperm samples incubated with cryopreservatives for 18 h at 20 degrees C did not show an increase in the percentage of AR in samples incubated with glycerol and ethylene glycol, while a significant variation was observed in the sample incubated with dimethyl sulphoxide (P < 0.001). Additional incubation for 3 h at 37 degrees C significantly increased the AR only in the sample incubated with glycerol (P < 0.001). Acrosome preservation is essential in the fertilization process and the evaluation of acrosome reaction induction by low temperature test was satisfactory. This test proves that ethylene glycol presents a greater protective effect on the acrosome preservation of human spermatozoa. PMID- 17081177 TI - Transmission electron microscopy, immunocytochemical and fluorescence in situ hybridisation studies in a case of 100% necrozoospermia: case report. AB - We present the ultrastructural, functional and chromosomal analyses of sperm from an infertile man with a normal karyotype and 100% necrozoospermia. Tests for microbial infection showed the presence of Escherichia coli in seminal and urethral fluid. Semen analysis was performed concomitantly with the infection and repeated twice after the therapy, in the absence of infection, to control the possible recovery of necrozoospermia. Morphological sperm evaluation was performed by light, fluorescent and electron microscopy; meiotic segregation was examined by fluorescence in situ hybridisation (FISH) technique. Polymerase chain reaction (PCR) was carried out on DNA from peripheral blood lymphocytes to analyse partial sequences of the Akap4 and Akap3 genes. AKAP3 protein is synthesised in round spermatids, incorporated into the fibrous sheath and is involved in organising the basic structure of the fibrous sheath. AKAP4 protein is incorporated late in spermatid development and plays a major role in completing fibrous sheath assembly. The whole sperm population was immotile and non-viable. The ultrastructural characteristics of sperm necrosis were identified. Immunostaining of tubulin and AKAP4 proteins was negative in sperm tails. PCR did not reveal any deletions in the investigated regions. FISH sperm analysis highlighted an altered meiotic segregation. After recovery from infection, 100% necrozoospermia persisted and chromosomal sperm aneuploidies were still present. PMID- 17081178 TI - Late termination of pregnancy: law, policy and decision making in four English fetal medicine units:. AB - OBJECTIVE: UK abortion law allows terminations for fetal abnormality without gestational limit. This study aimed to understand the decision-making experiences of fetal medicine professionals working within this legal framework. DESIGN: Qualitative study using semistructured interviews. SETTING: Four English fetal medicine units. SAMPLE: Fifteen doctors and midwives working in fetal medicine units and the Director of a related voluntary sector group. METHODS: Thematic analysis of transcribed interviews. MAIN OUTCOME MEASURES: Attitudes to abortion legislation; how decisions are made about the offer of late abortion and feticide. RESULTS: Fetal medicine specialists acknowledged the difficulties of ensuring that they worked within the law and within their own ethical frameworks when making decisions about offering terminations after viability. Practice regarding which abnormalities meet the legal criteria appeared to be governed largely by consensus between colleagues within their own and other units and in discussion with other specialists. Study participants reported individual differences about abnormalities where they personally would not wish to be involved in a termination, and also noted a shift in general attitudes over time as to conditions that meet the legal criteria. A proscribed list was believed to be both unworkable, given the variability in diagnoses and unhelpful, leading to reduced patient care. CONCLUSIONS: Research is needed to monitor attitudes to, and interpretation of, UK abortion legislation, which permits termination after a late diagnosis of fetal abnormality without gestational limit. If attitudes are changing, it is important to understand why, and what the consequences will be for parents and for health professionals. PMID- 17081179 TI - Mercury concentration and fish consumption in Taiwanese pregnant women. AB - OBJECTIVE: The aim of this study was to assess the relationship between fish consumption and total mercury concentration in maternal blood, umbilical cord blood, and placenta tissue of pregnant women in Taiwan. DESIGN: Cross-sectional study. SETTING: A medical centre in Taipei, Taiwan. SAMPLE: Sixty-five pregnant women delivered between July 2004 and March 2005. METHODS: We administered a questionnaire to each woman in the third trimester and collected blood samples and placenta tissue after delivery. Mercury concentrations in the maternal blood, cord blood and placenta tissue were measured using mercury analyser (Hiranuma HG 310, Hitachi, Japan). A dietitian calculated the quantity of fish consumed from the questionnaire. MAIN OUTCOME MEASURES: The total mercury concentration in maternal blood, cord blood and placenta tissue. RESULTS: The mean total mercury concentration in maternal blood, cord blood and placenta tissue was 9.1 +/- 0.40 microgram/l, 10.0 +/- 0.55 microgram/l and 19.2 +/- 1.8 ng/g, respectively. Eighty-nine percent of the maternal blood mercury concentrations exceeded the US National Research Council recommended value of 5.8 microgram/l. Fish consumption while pregnant correlated significantly with maternal blood and cord blood mercury concentrations. CONCLUSIONS: Total mercury concentrations of maternal blood, cord blood and placenta tissue commonly exceeded recommended values, and were higher in women who ate fish more than three times a week while pregnant. PMID- 17081180 TI - Women's experience of decision making about mode of delivery after a previous caesarean section: the role of health professionals and information about health risks. AB - OBJECTIVE: To explore women's experiences of decision making about mode of delivery after previous caesarean section. DESIGN: A qualitative interview study. SETTING: Two city maternity units in southwest England and Eastern Scotland. SAMPLE: Twenty-one women who had recently delivered a baby and whose previous child was delivered by caesarean section. METHODS: Semi-structured interviews analysed using the framework approach. MAIN OUTCOME MEASURES: Women's views on the influence of uncertainty on decision making, issues concerning information provision and decision-making roles. RESULTS: Experiences of decision making varied considerably. Some women were certain about choosing either vaginal birth after caesarean or repeat elective caesarean section, others were very uncertain and for some this uncertainty persisted after the birth. Information was most commonly provided by hospital doctors (mainly consultants) and more often related to procedural issues rather than possible health risks and benefits. Women felt they had to actively seek information rather than it being provided routinely. Most women were able to make their own decision about mode of delivery. Health professionals generally took a supportive role whichever mode of delivery was chosen. Although many women were comfortable with this approach, some felt they would have liked more guidance. CONCLUSION: On the whole, women experienced having control over the decision about planned mode of delivery. For many, making this decision was difficult and for some it was the cause of prolonged anxiety. Women were often making the decision without being provided with comprehensive and specific information about possible health risks and benefits. We are currently conducting a randomised controlled trial to investigate whether access to a decision aid is beneficial to women in this situation. PMID- 17081181 TI - Misoprostol compared with prostaglandin E2 for labour induction in women at term with intact membranes and unfavourable cervix: a systematic review. AB - BACKGROUND: Misoprostol is a commonly used prostaglandin to induce labour. A potential risk of induction, however, is caesarean delivery, especially in women with an unfavourable cervix. OBJECTIVES: To evaluate the use of misoprostol, compared with prostaglandin E2 (PgE2), for labour induction in women at term with an unfavourable cervix and intact membranes. SEARCH STRATEGY: PubMed, Medline, EMBASE and the Cochrane Library were searched for articles published in any language from January 1987 to December 2005, using the keywords 'misoprostol', 'labour/labor' and 'induction'. SELECTION CRITERIA: We identified randomised trials of women at term (> or =37 weeks of gestation) with intact membranes and unfavourable cervix, undergoing labour induction with misoprostol, orally, vaginally, sublingually or buccally, compared with PgE2 vaginally or intracervically. DATA COLLECTION AND ANALYSIS: Caesarean delivery was the primary outcome, with tachysystole and hyperstimulation as secondary outcomes. The primary analysis compared any misoprostol with any PgE2 for all women, with a subgroup analysis for nulliparous women. Secondary analyses compared different routes and doses of misoprostol (oral or vaginal and 25 microgram or >25 microgram) and PgE2 (intracervical or vaginal). Relative risks (RR) and 95% confidence intervals (CI) were calculated using random effects models. Main results Fourteen of 611 articles identified met the criteria for systematic review, with three providing information for nulliparous women. There was no difference in the risk of caesarean delivery between misoprostol and PgE2 groups (RR = 0.99, 95% CI = 0.83-1.17). Any misoprostol was associated with higher risks of tachysystole and hyperstimulation compared with any PgE2 (RR = 1.86, 95% CI = 1.01-3.43 and RR = 3.71, 95% CI = 2.00-6.88, respectively). There was a higher rate of vaginal delivery within 24 hours among all vaginal deliveries with any misoprostol compared with any PgE2 (RR = 1.14, 95% CI = 1.00-1.31), and among all deliveries, a lower rate of oxytocin use (RR = 0.71, 95% CI = 0.60-0.85) but a trend towards increased meconium staining was observed (RR = 1.22, 95% CI = 0.96 1.55). The use of misoprostol at starting dosages >25 microgram had similar findings to the primary analysis. Studies of lower misoprostol dosing (starting dose of 25 microgram) did not show any differences in the outcomes of interest, but the sample size of this secondary analysis was small (304 women, 155 receiving misoprostol). AUTHOR'S CONCLUSIONS: Although misoprostol in women at term with an unfavourable cervix and intact membranes was more effective than PgE2 in achieving vaginal delivery within 24 hours, misoprostol does not reduce the rate of caesarean delivery either in all women or in the subgroup of nulliparous women, and it increases the rates of tachysystole and hyperstimulation. Further studies of misoprostol using a starting dose of 25 microgram may be warranted. PMID- 17081182 TI - Maternal hyperhomocysteinaemia is a risk factor for congenital heart disease. AB - OBJECTIVE: To investigate the inter-relation between mother and infant homocysteine, folate and vitamin B12 status and the risk of a child with congenital heart disease (CHD). DESIGN: Case-control study. SETTING: Erasmus MC, University Medical Centre, Rotterdam, the Netherlands. POPULATION: Participants were 149 case-mothers and their children with CHD (n = 151) and 183 control mothers with their children (n = 175). METHODS: Approximately 17 months after the index-pregnancy maternal fasting, children's random venous blood samples were drawn to measure plasma total homocysteine, serum and red blood cell (RBC) folate, and serum vitamin B12 concentrations. Data were compared between cases and controls using the Mann-Whitney U test. The biochemical parameters were dichotomised according to the cutoff value of the 10th percentile of vitamin concentrations and the 90th percentile of homocysteine concentrations based on control data. Risk estimates for the association between CHD and the biochemical parameters were estimated in a logistic regression model. MAIN OUTCOME MEASURES: Medians (minimum-maximum) and odds ratios (OR) (95% confidence intervals [CI]). RESULTS: The OR (95% CI) of having a child with CHD was 2.9 (1.4-6.0) for maternal hyperhomocysteinaemia (>14.3 micromol/l). This finding is substantiated by a significant concentration-dependent risk (Ptrend = 0.004). Hyperhomocysteinaemic case-mothers showed significantly lower serum folate and vitamin B12 concentrations than normohomocysteinaemic case-mothers. Serum and RBC folate concentrations were significantly higher in case-children than that in control-children. CONCLUSIONS: Maternal hyperhomocysteinaemia is associated with an increased risk of CHD, partially due to low folate and vitamin B12 status. The folate status of children warrants further investigation. PMID- 17081183 TI - Virtual reality simulation training can improve technical skills during laparoscopic salpingectomy for ectopic pregnancy. AB - OBJECTIVES: To assess the first commercially available virtual reality (VR) simulator to incorporate procedural modules for training of inexperienced gynaecological surgeons to perform laparoscopic salpingectomy for ectopic pregnancy. DESIGN: Prospective cohort study. SETTING: Departments of surgery and gynaecology in central London teaching hospitals. SAMPLE: Thirty gynaecological surgeons were recruited to the study, and were divided into novice (<10 laparoscopic procedures), intermediate (20-50) and experienced (>100) groups. METHODS: All subjects were orientated to the VR simulator with a basic skills task, followed by performing ten repetitions of the virtual ectopic pregnancy module, in a distributed manner. MAIN OUTCOME MEASURES: Operative performance was assessed by the time taken to perform surgery, blood loss and total instrument path length. RESULTS: There were significant differences between the groups at the second repetition of the ectopic module for time taken (median 551.1 versus 401.2 versus 249.2 seconds, P = 0.001), total blood loss (median 304.2 versus 187.4 versus 123.3 ml, P = 0.031) and total instrument path length (median 17.8 versus 8.3 versus 6.8 m, P = 0.023). The learning curves of the experienced operators plateaued at the second session, although greater numbers of sessions were necessary for intermediate (seven) and novice (nine) surgeons to achieve similar levels of skill. CONCLUSIONS: Gynaecological surgeons with minimal laparoscopic experience can improve their skills during short-phase training on a VR procedural module. In contrast, experienced operators showed nonsignificant improvements. Thus, VR simulation may be useful for the early part of the learning curve for surgeons who wish to learn to perform laparoscopic salpingectomy for ectopic pregnancy. PMID- 17081184 TI - Is elective single embryo transfer a cost-effective alternative to double embryo transfer? AB - Elective single embryo transfer (eSET) is increasingly being considered as a means to reduce twin pregnancies associated with in vitro fertilisation treatment. However, it is important to consider the cost-effectiveness of alternative strategies when considering a change in policy. A review of the literature showed only five studies assessing both costs and consequences of strategies involving eSET compared with double embryo transfer. Several limitations in these studies prevent a definitive conclusion on the cost effectiveness of eSET being reached. Future economic evaluations need to compare strategies relevant to routine practice, include all relevant costs, measure and value longer term outcomes appropriately, and assess the cost-effectiveness of eSET across different subgroups of women. PMID- 17081185 TI - Consensus on the management of obstetric cholestasis: National UK survey. AB - An anonymous questionnaire was sent to the lead consultant obstetricians and the midwives in charge of the antenatal clinics of all maternity units in UK to determine their current views on the diagnosis and management of obstetric cholestasis (OC). The response rate was 60%. Management policies were broadly similar and consistent with those recommended in the current literature. However, the survey highlighted areas for improvement. Based on the findings of this survey, a consensus view of the management of OC in UK is achievable. PMID- 17081186 TI - First-line gemcitabine and carboplatin in advanced ovarian carcinoma: a phase II study. AB - OBJECTIVES: To evaluate tumour response rate and toxicities of gemcitabine and carboplatin in chemonaive subjects with advanced epithelial ovarian cancer. DESIGN: A phase II study. SETTING: Gynaecologic oncologic unit. POPULATION: Twenty chemonaive International Federation of Gynecology and Obstetrics (FIGO) stage IIIc and stage IV subjects with ovarian cancer. MAIN OUTCOME MEASURES: Tumour response, disease free and overall survival and toxicity. METHODS: Intravenous gemcitabine 1000 mg/m2 on day 1 and day 8 and carboplatin at area under centre (AUC) = 5 on day 1 were administered three weekly for six cycles. Subjects who received more than three cycles of chemotherapy were eligible for assessment of tumour response, while all the cycles of chemotherapy were assessed for toxicities. RESULTS: The mean age of the subjects was 57.3 years, and the median follow up was 38.7 months. Of the 18 eligible subjects analysed, 11 (61.1%) showed a complete clinical response, and the overall response rate was 83.3% (15/18). The median overall survival was 29.2 [95% (confidence interval) CI 22.8-35.6] months, and the median progression-free survival was 11.6 (95% CI 4.7 18.5) months. WHO grade 3 anaemia, neutropenia and thrombocytopenia was 7.6, 9.5 and 0%, respectively, on day 8, and 15.5, 12.2 and 15.5%, respectively, on day 15. Two subjects required a total of three hospital admissions for neutropenic sepsis, and two required five hospital admissions for platelet transfusion for severe thrombocytopenia. CONCLUSION: Chemonaive advanced ovarian cancer showed a high response rate to combined gemcitabine and carboplatin chemotherapy. The subjects developed moderate adverse reactions. Phase III study to evaluate the role of combined gemcitabine and carboplatin as first-line chemotherapy in ovarian cancer is warranted. PMID- 17081187 TI - Vaginal vault smears after hysterectomy for reasons other than malignancy: a systematic review of the literature. AB - BACKGROUND: Vaginal vault smears are used to detect persisting neoplasia of the lower genital tract after hysterectomy. Recent data suggest both widespread use and uncertain evidence of their effectiveness. OBJECTIVES: To identify and synthesise evidence on the use and effectiveness of vaginal vault smears and to assess the quality. SEARCH STRATEGY 'vault smear' OR 'vaginal vault smear' OR 'cervical vault smear' OR ('Hysterectomy') AND ('Follow up' OR 'Smear'). SELECTION CRITERIA Primary research, women who had a hysterectomy and were followed up by vault cytology. DATA COLLECTION AND ANALYSIS: Systematic search (eight electronic databases), supplemented by contact with experts and review of bibliographies. Two independent reviewers determined eligibility/validity and extracted data concerning test performance characteristics. Quality was assessed according to the established criteria. RESULTS: Of 441 unique references, only 19 were suitable. Quality of studies varied considerably and few were of 'high' methodological quality. Studies were geographically diverse, and were published over more than 40 years in 16 journals. From the higher scoring papers, there were 11 659 hysterectomies [6546, benign; 76, cervical intraepithelial neoplasia (CIN) I/CIN II; 5037, CIN III]. Proportions of abnormal vault smears and abnormal biopsies during follow up increased with worsening histology at hysterectomy (P < 0.0001 and P = 0.0001). There was only one report of vaginal cancer subsequent to hysterectomy for CIN and insufficient data to allow for reliable meta-analysis. CONCLUSIONS: Vault smears cause anxiety, consume resources and their value is largely unproven. Inconsistency of study design and limited methodological quality means that the value of vault smears could not be established. High quality research is required to ensure that the guidelines are evidence based. PMID- 17081188 TI - Folic acid knowledge and use in a multi-ethnic pregnancy cohort: the role of language proficiency. AB - OBJECTIVE: To investigate the role of language proficiency as determinant of folic acid knowledge and use in a multi-ethnic pregnancy cohort. DESIGN: Prospective cohort study. SETTING AND POPULATION: Pregnant women from Amsterdam attending obstetric care for their first antenatal visit. Number approached: 12,373 women, response rate: 67% (8266 women aged 14-49 years). Ethnicity was based on the country of birth: the Netherlands, Surinam, Antilles, Turkey, Morocco, Ghana, other non-Western and other Western countries. MAIN OUTCOME MEASURES: Knowledge about and use of folic acid supplements in pregnancy as elicited in a multilingual questionnaire, as well as determinants of these in ethnic groups separately. RESULTS: Both periconceptional folic acid use and knowledge were significantly lower among Ghanaian, Moroccan, Turkish, and other non-Western women than among women born in the Netherlands or other Western countries. Language proficiency in Dutch was a major determinant of knowledge in all the ethnic groups with a mother tongue other than Dutch [adjusted odds ratios (OR): Western 3.2, non-Western (all countries combined) 7.5], while educational attainment was of secondary importance. Knowledge in turn was the strongest determinant of use (adjusted OR: Western 17.4, non-Western 27.0). CONCLUSIONS: Periconceptional folic acid supplement use among women born in non-Dutch-speaking non-Western countries is low, reflecting a lack of knowledge that is determined by the inability to speak and understand the language of the country of residence. Measures to tackle this problem include the provision of linguistically appropriate information via ethnic health advisors, and language courses integrating health education for immigrants. PMID- 17081190 TI - Cell biology of plant-oomycete interactions. AB - The last 4 years have seen significant advances in our understanding of the cellular processes that underlie the infection of plants by a range of biotrophic and necrotrophic oomycete pathogens. Given that oomycete and fungal pathogens must overcome the same sets of physical and chemical barriers presented by plants, it is not surprising that many aspects of oomycete infection strategies are similar to those of fungal pathogens. A major difference, however, centres on the role of motile oomycete zoospores in actively moving the pathogen to favourable infection sites. Recent studies have shown that the plant defence response to invading oomycetes is similar to that mounted against fungi, but biochemical differences between oomycete and fungal surface molecules must have implications for plant recognition of and defence against oomycete pathogens. The aim of this short review is to provide a cell biological framework within which emerging data on the molecular basis of oomycete-plant interactions may be placed. PMID- 17081191 TI - Activation of defence reactions in Solanaceae: where is the specificity? AB - When a potential pathogen attempts to infect a plant, biochemical and molecular communication takes place and leads to the induction of plant defence mechanisms. In the case of efficient defence, visible symptoms are restricted and the pathogen does not multiply (incompatible interaction); when defence is inefficient, the plant becomes rapidly infected (compatible interaction). During the last 30 years, a growing body of knowledge on plant-pathogen interactions has been gathered, and a large number of studies investigate the induction of various plant defence reactions by pathogens or by pathogen-derived compounds. However, as most papers focus on incompatible interactions, there is still a lack of understanding about the similarities and differences between compatible and incompatible situations. This review targets the question of specificity in Solanaceae-pathogen interactions, by comparing defence patterns in plants challenged with virulent or avirulent pathogens (or with pathogen-associated molecular patterns from these). A special emphasis is made on analysing whether defence reactions in Solanaceae depend primarily on the type of elicitor, on the plant genotype/species, or on the type of interaction (compatible or incompatible). PMID- 17081192 TI - Vesicle trafficking in plant immune responses. AB - In plants, perception of pathogen-associated molecular patterns at the surface is the first line of defence in cellular immunity. This review summarizes recent evidence of the involvement of vesicle trafficking in the plant's immune response against pathogens. I first discuss aspects of ligand-stimulated receptor endocytosis. The best-characterized pattern-recognition receptor (PRR), FLS2, is a transmembrane leucine-rich repeat receptor kinase that recognizes bacterial flagellin. FLS2 was recently shown to undergo internalization upon activation with its cognate ligand. An animal PRR, TLR4 that mediates perception of bacterial-derived lipopolysaccharides, similarly exhibits ligand-stimulated endocytosis. The second focus is N-ethylmaleimide-sensitive factor adaptor protein receptor (SNARE)-mediated immunity involving syntaxins and their cognate partners. One of the genes involved in basal immunity in Arabidopsis, PEN1, encodes a syntaxin that focally accumulates at fungal penetration sites, raising the possibility that induced exocytosis is important for active defence. Pathogen triggered endocytic and exocytic processes have to be balanced to ensure host cell homeostasis. Thus, understanding how phytopathogens have evolved strategies to exploit host cell vesicle trafficking to manipulate immune responses is currently an area of intense study. PMID- 17081193 TI - Rotavirus impairs the biosynthesis of brush-border-associated dipeptidyl peptidase IV in human enterocyte-like Caco-2/TC7 cells. AB - Rotavirus is the leading cause of severe dehydrating diarrhoea in infants and young children worldwide. This virus infects mature enterocytes in the small intestine, and induces structural and functional damage. In the present study, we have identified a new mechanism by which rotavirus impairs a brush border associated intestinal protein. We show that infection of enterocyte-like Caco 2/TC7 cells by rhesus monkey rotavirus (RRV) impairs the biosynthesis of dipeptidyl peptidase IV (DPP IV), an important hydrolase in the digestion of dietary proline-rich proteins. We show that the enzyme activity of DPP IV was reduced, and that rearrangements of the protein occurred at the apical domain of the RRV-infected cells. Using pulse-chase experiments and cell surface immunoprecipitation, we have demonstrated that RRV infection did not affect the stability or apical targeting of DPP IV, but did induce a dramatic decrease in its biosynthesis. Using quantitative RT-PCR, we showed that RRV had no effect on the level of expression of DPP IV mRNA, suggesting that the observed decrease in the biosynthesis of the protein is related to an effect of the virus at the translational level. PMID- 17081194 TI - The GacA global regulator of Vibrio fischeri is required for normal host tissue responses that limit subsequent bacterial colonization. AB - Harmful and beneficial bacterium-host interactions induce similar host-tissue changes that lead to contrasting outcomes of association. A life-long association between Vibrio fischeri and the light organ of its host Euprymna scolopes begins when the squid collects bacteria from the surrounding seawater using mucus secreted from ciliated epithelial appendages. Following colonization, the bacterium causes changes in host tissue including cessation of mucus shedding, and apoptosis and regression of the appendages that may limit additional bacterial interactions. We evaluated whether delivery of morphogenic signals is influenced by GacA, a virulence regulator in pathogens, which also influences squid-colonization by V. fischeri. Low-level colonization by a GacA mutant led to regression of the ciliated appendages. However, the GacA mutant did not induce cessation of mucus shedding, nor did it trigger apoptosis in the appendages, a phenotype that normally correlates with their regression. Because apoptosis is triggered by lipopolysaccharide, we examined the GacA mutant and determined that it had an altered lipopolysaccharide profile as well as an increased sensitivity to detergents. GacA-mutant-colonized animals were highly susceptible to invasion by secondary colonizers, suggesting that the GacA mutant's inability to signal the full programme of light-organ responses permitted the prolonged recruitment of additional symbionts. PMID- 17081195 TI - Virulence of Porphyromonas gingivalis is altered by substitution of fimbria gene with different genotype. AB - Porphyromonas gingivalis is a periodontal pathogen whose fimbriae are classified into six genotypes based on the diversity of the fimA genes encoding each fimbria subunit. It was suggested that P. gingivalis strains with type II fimbriae were more virulent than type I strains. For the present study, we generated the mutants in which fimA was substituted with different genotypes to study virulence of type II fimbriae. Using plasmid vectors, fimA of ATCC33277 (type I strain) was substituted with type II fimA, and that of OMZ314 (type II strain) with type I fimA. The substitution of type I fimA with type II enhanced bacterial adhesion/invasion to epithelial cells, whereas substitution with type I fimA resulted in diminished efficiency. Following bacterial invasion, type II clones swiftly degraded cellular paxillin and focal adhesion kinase, and inhibited cellular migration, whereas type I clones and DeltafimA mutants did not. BIAcore analysis demonstrated that type II fimbriae possess greater adhesive abilities for their receptor alpha5beta1-integrin than those of type I. In a mouse abscess model, the type II clones significantly induced serum IL-1beta and IL-6, as well as other infectious symptoms. These results suggest that type II fimbriae are a critical determinant of P. gingivalis virulence. PMID- 17081196 TI - Human recombinant prolidase from eukaryotic and prokaryotic sources. Expression, purification, characterization and long-term stability studies. AB - Prolidase is a Mn(2+)-dependent dipeptidase that cleaves imidodipeptides containing C-terminal proline or hydroxyproline. In humans, a lack of prolidase activity causes prolidase deficiency, a rare autosomal recessive disease, characterized by a wide range of clinical outcomes, including severe skin lesions, mental retardation, and infections of the respiratory tract. In this study, recombinant prolidase was produced as a fusion protein with an N-terminal histidine tag in eukaryotic and prokaryotic hosts and purified in a single step using immobilized metal affinity chromatography. The enzyme was characterized in terms of activity against different substrates, in the presence of various bivalent ions, in the presence of the strong inhibitor Cbz-Pro, and at different temperatures and pHs. The recombinant enzyme with and without a tag showed properties mainly indistinguishable from those of the native prolidase from fibroblast lysate. The protein yield was higher from the prokaryotic source, and a detailed long-term stability study of this enzyme at 37 degrees C was therefore undertaken. For this analysis, an 'on-column' digestion of the N-terminal His tag by Factor Xa was performed. A positive effect of Mn(2+) and GSH in the incubation mixture and high stability of the untagged enzyme are reported. Poly(ethylene glycol) and glycerol had a stabilizing effect, the latter being the more effective. In addition, no significant degradation was detected after up to 6 days of incubation with cellular lysate. Generation of the prolidase in Escherichia coli, because of its high yield, stability, and similarity to native prolidase, appears to be the best approach for future structural studies and enzyme replacement therapy. PMID- 17081197 TI - Septaplex PCR assay for rapid identification of Vibrio cholerae including detection of virulence and int SXT genes. AB - In this study, we describe a septaplex PCR assay for rapid identification of Vibrio cholerae including detection of the virulence and intsxt genes. Conditions were optimized to amplify fragments of ISRrRNA (encoding for 16S-23S rRNA gene, Intergenic spacer regions), O1rfb (O1 serogroup specific rfb), O139rfb (O139 serogroup specific rfb), ctxA (cholera toxin subunit A), tcpA (toxin coregulated pilus), and intsxt (sxt integron) simultaneously in a single PCR. The septaplex PCR was evaluated using 211 strains of V. cholerae and six water samples for in situ testing. PCR results were correlated with genotype data obtained by individual PCR and slot-blot assays. The one-step PCR described here can be used to identify V. cholerae accurately and rapidly. Also, the virulence and intsxt genes can be simultaneously detected, providing a useful method for monitoring pathogenic, intsxt-positive and nonpathogenic, intsxt-negative V. cholerae serogroups both in the environment and clinical settings. PMID- 17081198 TI - Oral immunization of BALB/c mice by intragastric delivery of Streptococcus gordonii-expressing Giardia cyst wall protein 2 decreases cyst shedding in challenged mice. AB - Giardia lamblia (Giardia duodenalis or Giardia intestinalis) is a protozoan parasite of vertebrates with broad host specificity. Specific antibodies directed against cyst antigens can interfere with the cyst wall-building process. In this study, we engineered Streptococcus gordonii to express a 26 kDa fragment of cyst wall protein 2 (CWP2), containing a relevant B cell epitope, on the cell surface. This is the first report of S. gordonii expressing a protein of parasite origin. As S. gordonii was intended for intestinal delivery of CWP2, it was determined that this oral commensal bacterium is able to persist in the murine intestine for 30 days. Immunization with recombinant streptococci expressing the 26 kDa fragment resulted in higher antibody levels. Specific anti-CWP2 IgA antibodies were detected in fecal samples and anti-CWP2 IgG antibodies were detected in serum demonstrating the efficacy of S. gordonii for intragastric antigen delivery. In a pilot challenge experiment, immunized mice demonstrated a significant 70% reduction in cyst output. PMID- 17081199 TI - Effects of polymicrobial communities on host immunity and response. AB - Microorganisms grow as members of microbial communities in unique niches, such as the mucosal surfaces of the human body. These microbial communities, containing both commensals and opportunistic pathogens, serve to keep individual pathogens 'in check' through a variety of mechanisms and complex interactions, both between the microorganisms themselves and the microorganisms and the host. Recent studies shed new light on the diversity of microorganisms that form the human microbial communities and the interactions these microbial communities have with the host to stimulate immune responses. This occurs through their recognition by dendritic cells or their ability to induce differential cytokine and defensin profiles. The differential induction of defensins by commensals and pathogens and the ability of the induced defensins to interact with the antigens from these microorganisms may attenuate proinflammatory signaling and trigger adaptive immune responses to microbial antigens in a multistep process. Such an activity may be a mechanism that the host uses to sense what is on its mucosal surfaces, as well as to differentiate among commensals and pathogens. PMID- 17081200 TI - ICD arrhythmia detection and discrimination: are we there yet? PMID- 17081201 TI - Atrial fibrillation: much heat, a little light. PMID- 17081202 TI - How to make a submuscular pocket. PMID- 17081203 TI - A novel pacing maneuver to localize focal atrial tachycardia. AB - BACKGROUND: Although focal atrial tachycardias cannot be entrained, we hypothesized that atrial overdrive pacing (AOP) can be an effective adjunct to localize the focus of these tachycardias at the site where the post-pacing interval (PPI) is closest to the tachycardia cycle length (TCL). METHODS: Overdrive pacing was performed in nine patients during atrial tachycardia, and in a comparison group of 15 patients during sinus rhythm. Pacing at a rate slightly faster than atrial tachycardia in group 1 and sinus rhythm in group 2 was performed from five standardized sites in the right atrium and coronary sinus. The difference between the PPI and tachycardia or sinus cycle length (SCL) was recorded at each site. The tachycardia focus was then located and ablated in group 1, and the atrial site with earliest activation was mapped in group 2. RESULTS: In both groups the PPI-TCL at the five pacing sites reflected the distance from the AT focus or sinus node. In group 1, PPI-TCL at the successful ablation site was 11 +/- 8 msec. In group 2, PPI-SCL at the site of earliest atrial activation was 131 +/- 37 msec (P < 0.001 for comparison). In groups 1 and 2, calculated values at the five pacing sites were proportional to the distance from the AT focus or sinus node, respectively. CONCLUSIONS: The PPI-TCL after-AOP of focal atrial tachycardia has a direct relationship to proximity of the pacing site to the focus, and may be clinically useful in finding a successful ablation site. PMID- 17081204 TI - Electrocardiogram during simultaneous biventricular pacing and cardiomyoplasty stimulation. PMID- 17081205 TI - Characteristics of patients undergoing atrial fibrillation ablation: trends over a seven-year period 1999-2005. AB - BACKGROUND: Catheter ablation procedures for treating atrial fibrillation (AF) have dramatically increased since triggers of AF were first described in 1998. OBJECTIVE: We explored changes in patient characteristics in patients referred for catheter ablation of AF over a seven-year period from 1999 through 2005. METHODS: Patient characteristics were examined for all patients undergoing AF ablation from 1999 through 2005 at the University of Pennsylvania Health System (UPHS). The gender of patients undergoing ablation was also compared with outpatients seen at UPHS with a primary diagnosis of AF. RESULTS: From 1999 to 2005 the number of patients undergoing ablation has increased steadily, from 29 patients in 1999 to 265 patients in 2005 (P < 0.01). Patients have become older (47 to 56 years; P < 0.01), with more persistent or permanent AF (17% to 45%; P < 0.01), larger left atrial size (4.0 to 4.4 cm; P < 0.01), and fewer antiarrhythmic drugs used prior to ablation (3.9 to 2.0 drugs; P < 0.01). Patients undergoing ablation have been predominantly male, with a significantly higher male prevalence than patients seen in the UPHS outpatient primary care clinic with AF (77% vs 59% male; P < 0.001). CONCLUSIONS: Patients undergoing AF ablation from 1999 to 2005 are older, with larger left atrial size, more persistent/permanent AF, and fewer prior antiarrhythmic agents used. Compared with the gender-specific rates of AF in the population, the majority of patients referred for ablation are men, suggesting a referral bias against this invasive procedure for women. These findings are important for interpreting the outcome of ablation in the current era, and for designing prospective randomized trials. PMID- 17081206 TI - Palpitations with alternating heart rate: what is the mechanism? PMID- 17081207 TI - Repeat transseptal catheterization after ablation for atrial fibrillation. AB - INTRODUCTION: A substantial number of patients require a second left atrial procedure after ablation for atrial fibrillation (AF), either for left atrial flutter or recurrent AF. The success and complication rates of repeat transseptal catheterization in these patients are unknown. The aim of this study was to determine the difficulty and/or success rates of repeat transseptal catheterization after left atrial ablation for AF. METHODS AND RESULTS: Consecutive patients undergoing repeat left atrial procedures after ablation for AF over a 1-year period were enrolled. Difficulties with, success rates, and complications of the first and second transseptal catheterizations were recorded. Sixteen patients underwent a repeat transseptal catheterization. Of the 4 in whom the first procedure was performed with an ablation catheter across a patent foramen ovale (PFO), 3 required a transseptal puncture for their repeat procedure. The remaining 12 underwent transseptal puncture without difficulty for their first procedure, and, despite the same operators for each patient, the repeat transseptal was noted to be difficult in 5. Of those 5, the transseptal puncture was unsuccessful due to increased interatrial septal thickness in 2 patients. One repeat transseptal attempt was aborted after posterior right atrial puncture with the transseptal needle occurred, attributed to distorted interatrial septal anatomy not observed prior to the first case. CONCLUSIONS: Compared with the first procedure, repeat transseptal catheterization after ablation for AF, whether initially performed across a PFO or via a transseptal puncture, is more difficult, less often successful, and potentially associated with more complications. PMID- 17081208 TI - The Managed Ventricular pacing versus VVI 40 Pacing (MVP) Trial: clinical background, rationale, design, and implementation. AB - BACKGROUND: Implantable cardioverter defibrillators (ICDs) reduce mortality among appropriately selected patients who have had or are at risk for life-threatening ventricular arrhythmia. Right ventricular apical (RVA) pacing has been implicated in worsening heart failure and death. The optimal pacemaker mode for bradycardia support while minimizing unnecessary and potentially harmful RVA pacing has not been determined. METHODS: The Managed Ventricular pacing vs. VVI 40 Pacing Trial (MVP) is a prospective, multicenter, randomized, single-blind, parallel, controlled clinical trial designed to establish whether atrial-based dual-chamber managed ventricular pacing mode (MVP) is equivalent or superior to back-up only ventricular pacing (VVI 40) among patients with standard indications for ICD therapy and no indication for bradycardia pacing. The MVP Trial is designed with 80% power to detect a 10% reduction in the primary endpoint of new or worsening heart failure or all-cause mortality in the MVP-treated group. Approximately 1,000 patients at 80 centers in the United States, Canada, Western Europe, and Israel will be randomized to MVP or VVI 40 pacing after successful implantation of a dual-chamber ICD. Heart failure therapies will be optimized in accordance with evidence-based guidelines. Prespecified secondary endpoints will include ventricular arrhythmias, atrial fibrillation, new indication for bradycardia pacing, health-related quality of life, and cost effectiveness. Enrollment began in October 2004 and concluded in April 2006. The study will be terminated upon recommendation of the Data Monitoring Committee or when the last patient enrolled and surviving has reached a minimum 2 years of follow-up. CONCLUSION: The MVP Trial will meet the clinical need for carefully designed prospective studies to define the benefits of atrial-based dual-chamber minimal ventricular pacing versus single-chamber ventricular pacing in conventional ICD patients. PMID- 17081209 TI - Electroanatomic remodeling of the left atrium in patients undergoing repeat pulmonary vein ablation: mechanistic insights and implications for ablation. AB - INTRODUCTION: There is limited information describing late changes in the electroanatomic characteristics of the left atrium (LA) associated with recurrence after an anatomical circumferential pulmonary vein ablation (CPVA) for atrial fibrillation (AF). METHODS AND RESULTS: Forty-seven patients (57 +/- 8 years) undergoing a repeat ablation after CPVA were included. Using an electroanatomic mapping system, we measured the bipolar voltage by averaging points in the pulmonary vein (PV)-LA junction and four other LA sites. Conduction velocity and AF cycle length (AFCL) were also measured and the results are compared with the first procedure. After an initial decrease observed at the end of the first procedure, voltage and conduction velocity returned to intermediate values in all LA sites, with lower voltage at the LIPV antrum (P = 0.004), and lower conduction velocity across the LIPV and RSPV (P < 0.001). Conduction gaps were more prevalent at the septal aspect of the right PV encircling lines (85%), between the left atrial appendage (LAA) and the LSPV (70%) and lines at the posterior wall (71%). There was a nonsignificant increase in AFCL, with a more widespread distribution of organized electrograms (32.4% vs 46.6%). CONCLUSION: Recurrence after CPVA is associated with a reverse process of voltage and conduction velocity increase across ablated areas, especially the PV-LA junction, and is related to the presence of conduction gaps, which are distributed mostly at the septal aspect of the lines encircling the right PVs and at the LAA-LSPV area. Organization of atrial electrograms seen during AF ablation is maintained at a repeat procedure. PMID- 17081210 TI - Reversal of left ventricular dysfunction following ablation of atrial fibrillation. AB - BACKGROUND: Evaluation of ventricular rate control in atrial fibrillation (AF) can be difficult, and the presence of an AF-induced ventricular cardiomyopathy due to intermittent poor rate control or other causes may be underestimated. The outcome with AF ablation in patients with a decreased left ventricular ejection fraction (LVEF) may provide insight into this important clinical issue. OBJECTIVE: To determine the effect of pulmonary vein isolation on LVEF in patients with AF and decreased LVEF (< or = 50%). METHODS: Ablation consisted of proximal isolation of arrhythmogenic pulmonary veins (PVs) and elimination of non PV triggers. LVEF was determined within 24 hours after ablation and again at up to 6 months follow-up. Transtelephonic monitoring was performed routinely for 2-3 weeks prior to ablation, at 6 weeks, and 6 months post and with symptoms following ablation. AF control was defined as freedom from AF or marked (>90%) reduction in AF burden on or off previously ineffective antiarrhythmic medication. RESULTS: AF ablation was performed in 366 patients and 67 (18%) patients had decreased LV function with a mean LVEF of 42 +/- 9%. An average of 3.4 +/- 0.9 PVs were isolated. AF control in the depressed LVEF group compared favorably with the normal EF group (86% vs. 87% P = NS), although more redo procedures were required (1.6 +/- 0.8 vs 1.3 +/- 0.6 procedures; P < or = 0.05). Only 15 of 67 patients (22%) with decreased LVEF had shown tachycardia (>100 bpm) on repeated preablation ECG recordings during AF. In the decreased LVEF group, the LVEF increased from 42 +/- 9% to 56 +/- 8% (P < 0.001) after ablation. CONCLUSIONS: Patients with AF and decreased LVEF undergoing AF ablation have similar success to patients with normal LVEF and have improvement in LVEF after ablation. These results suggest the presence of a reversible AF-induced ventricular cardiomyopathy in many patients with AF and depressed LV function. The presence of under-recognized and reversible cardiomyopathy even when tachycardia is not persistent is important to recognize. PMID- 17081211 TI - Cycle length alternation during pacing. PMID- 17081212 TI - Newly detected atrial fibrillation following dual chamber pacemaker implantation. AB - INTRODUCTION: Pacemaker (PPM)-detected atrial high-rate episodes (AHREs) of even 5-minute duration may identify patients at increased risk for stroke and death. In this study, we sought to determine the incidence of newly detected atrial fibrillation (AF defined as an AHRE > or = 5 minutes) in patients following dual chamber PPM implantation and to define the clinical predictors of developing AF. METHODS AND RESULTS: We evaluated 262 patients (142 male; age 74 +/- 12 years) without documented AF who underwent PPM implantation for sinus node dysfunction (n = 122) or atrioventricular block (n = 140). Information regarding patient demographics, cardiovascular diseases, and medication history was obtained. The cumulative percentages of ventricular pacing as well as the frequency, duration, and time to first episode of an AHRE were also determined. During follow-up of 596 +/- 344 days, an AHRE > or = 5 minutes was detected in 77 (29%) patients. Of these, 47 (61%) patients had an AHRE > or = 1 hour, 22 (29%) patients had an AHRE > or = 1 day, and 12 (16%) patients had an AHRE > or = 1 week. An AHRE > or = 5 minutes was seen in 24% and 34% of patients at 1 year and 2 years, respectively. Among patients with sinus node dysfunction, > or = 50% cumulative ventricular pacing was the only significant predictor of an AHRE > or = 5 minutes (HR 2.2; CI 1.0-4.7; P = 0.04). CONCLUSIONS: Within 1 year of PPM implantation, AF is detected in 24% of patients without history of AF. In patients with sinus node dysfunction, > or = 50% cumulative right ventricular pacing is associated with a 2-fold increase in risk of developing AF. PMID- 17081213 TI - The electroanatomic characteristics of the cavotricuspid isthmus: implications for the catheter ablation of atrial flutter. AB - INTRODUCTION: Radiofrequency ablation (RFA) of typical AFL is sometimes difficult because of the poor electroanatomic approach to the cavotricuspid isthmus (CTI). The aim of this study was to correlate the anatomy of the CTI between contact mapping (NavX) and right atrial angiography (RAG), and to investigate the impact of the electroanatomic characteristics of the CTI on the RFA of typical atrial flutter (AFL). METHODS: One hundred patients with typical AFL undergoing RFA were studied. The image-guided group consisted of 50 consecutive patients with the guidance of NavX. NavX geometry and RAG were performed to investigate the morphology of the CTI. The bipolar voltages of the CTI were collected during sinus rhythm by a NavX. The control group consisted of 50 consecutive patients with the guidance of conventional fluoroscopy. RESULTS: There was a good correlation between the angiography and NavX for the anatomy of the CTI. The pouch type had a longer length of CTI than the flat type (33.4 +/- 5.0 vs 22.6 +/ 8.4 mm, P < 0.0001) and deeper depth than the concave type (6.5 +/- 2.2 vs 3.7 +/- 0.8 mm, P < 0.0001) on the angiography. The pouch-type CTI had a longer ablation time and larger pulses of RFA than the other two types. The control group had a longer ablation time, fluoroscopy time, and larger pulses of RFA than image-guided group. CONCLUSIONS: The 3-D mapping system provided a good reconstruction of CTI, which may help in the RFA in patients with a complex anatomy of the CTI. PMID- 17081214 TI - Incidence and predictors of very late recurrence of atrial fibrillation after ablation. AB - INTRODUCTION: Radiofrequency catheter ablation can effectively treat patients with refractory atrial fibrillation (AF). Very late AF recurrence (> or = 12 months post-ablation) is uncommon and may represent a unique patient cohort. METHODS AND RESULTS: A nested case-control study was performed in the cohort who underwent AF ablation at the University of Pennsylvania to characterize patients who develop very late AF recurrence after ablation. The procedure consisted of isolation of pulmonary veins (PVs) demonstrating triggers and elimination of non PV triggers initiating AF. Twenty-seven (7.9%) patients with very late recurrence were compared to 219 patients without recurrence and > or = 12 months of follow up. The mean age was 54.6 +/- 11.3 years and 79% were men. Very late recurrence patients more likely weighed >200 lbs (70% vs 55%, P = 0.01); during initial ablation had fewer PVs isolated (2.8 +/- 1.1 vs 3.3 +/- 1.0, P = 0.03); and were less likely to have right inferior PV isolation (37% vs 61%, P = 0.02), less likely to have isolation of all PVs (30% vs 56%, P = 0.01), and more likely to have non-PV triggers (30% vs 11% OR 3.4(95% CI, 1.3-8.7), P = 0.01). PV reconnectivity and new triggers were found in the majority of patients with very late recurrence of AF who underwent repeat ablation. CONCLUSION: Very late recurrence of AF more likely occurred in patients >200 lbs who demonstrated non PV triggers and did not undergo right inferior PV isolation. The majority of patients undergoing repeat ablation for very late recurrence demonstrated PV reconnectivity and new non-PV and PV triggers not observed during the initial ablation. PMID- 17081215 TI - Preservation of existing inflatable penile prosthesis pump mechanism during scrotal surgery. AB - INTRODUCTION: There is little reported regarding the feasibility of scrotal surgery in patients with an existing penile prosthesis. We report a series of two patients with existing inflatable penile prostheses who underwent scrotal surgery for unrelated indications in which the prosthesis pump mechanism and tubing were preserved. In both instances, damage to the prosthetic pump and tubing were avoided. AIM: To demonstrate two instances where scrotal surgery was performed successfully in the setting of a pre-existing penile prosthesis. METHODS: Two cases of patients with existing penile implant who underwent scrotal surgery for pathology unrelated to the implant are reported. CONCLUSIONS: It is possible to perform scrotal surgery in a patient who has an existing penile prosthesis without injuring or infecting the prosthesis. PMID- 17081216 TI - Corporoscopic excavation of the fibrosed corpora cavernosa for penile prosethesis implantation: optical corporotomy and trans-corporeal resection, Shaeer's technique. AB - INTRODUCTION: Implantation of a penile prosthesis in cases of neglected or resistant ischemic priapism, or delayed re-implantation following prosthesis infection and extraction, is usually a difficult and risky procedure due to fibrosis of the corpora cavernosa. Among the common complications are perforation of the urethra, tunica albuginea, and infection. The complications are usually due to the use of blind force against resistance. AIM: We propose the techniques of Trans-Corporeal Resection and Optical Corporotomy as adjuvant measures for excavating the fibrosed corpora cavernosa under vision, without the use of force against resistance. METHODS: Six patients with diffuse fibrosis of the corpora cavernosa were operated on. The instruments and technique are the same as for optical urethrotomy and transurethral resection. Optical Corporotomy was started with, where the corpora are incised from within. After establishment of a satisfactory passage, Trans-Corporeal Resection followed to scrape the fibrous tissue. Implantation of penile prosthesis was completed as usual. The procedure was performed through 1.5 cm incision in the tunica albuginea. MAIN OUTCOME MEASURES: Length, girth, and straightness in the erect position, as well as the incidence of complications. RESULTS: Operative time was an average of 90 minutes. No difficulty was encountered during the procedure. No complications were noted through 1 year of follow-up. CONCLUSION: Optical Corporotomy and Trans-Corporeal Resection allow for force-free, visually monitored excavation of the fibrosed corpora cavernosa, aiming at safer penile prosthesis implantation. PMID- 17081217 TI - Dorsal vein injuries observed during penile exploration for suspected penile fracture. AB - INTRODUCTION: Penile fracture is a rare injury, bearing potential impairment of erectile function if not treated. Patients with clinical presentation of a penile fracture commonly undergo early surgical exploration with the intention to repair a tunica albuginea tear. AIM: We present a group of men who presented with a penile hematoma following trauma to the erect penis. Exploration revealed an intact tunica albuginea and a dorsal vein tear. METHODS: Eighteen men (mean age 38 years, range 20-55) presented with suspected penile fracture during an 8-year period. One man presented twice. Two of the patients were managed expectantly and the remaining 16 patients underwent 17 immediate surgical explorations. Explorations were performed under general anesthesia, using a circumferential subcoronal incision and degloving of the penile skin. The tunica albuginea of both penile sides as well as the penile urethra were examined for injuries. MAIN OUTCOME MEASURES: Medical records were retrospectively reviewed for etiology, symptoms, signs of physical examination, and information on findings of surgical exploration. Data on erectile function, medical treatment for erectile dysfunction, and penile curvature were obtained during follow-up. RESULTS: In nine of the 17 procedures the tunica albuginea was intact and the only pathological finding was a ruptured dorsal vein. One procedure was negative for both tunical and vascular injury. A tunical tear was detected in the remaining seven procedures. At a mean follow-up of 40 months (range 4-91), five patients required medical treatment for erectile dysfunction, including the two who were managed expectantly, two with a tunical tear, and one with a venous tear. CONCLUSIONS: Dorsal vein tears may mimic penile fracture. Suggestive findings following trauma to the erect penis prompted exploration for suspected tunica albuginea tear. In less than half of the men was the diagnosis of penile fracture established and treated at surgery. PMID- 17081218 TI - A reproducibility study of corpus cavernosum electromyography in young healthy volunteers under controlled conditions. AB - INTRODUCTION: Although the corpus cavernosum electromyography (CC-EMG) has been studied already for 16 years, doubts regarding its reproducibility have remained. AIM: To assess the reproducibility of CC-EMG under controlled conditions and the influence of confounding factors. METHODS: Three CC-EMG recording sessions were performed in 13 healthy young men under the same conditions. Furthermore, the effects of potentially confounding factors, such as intake of caffeine, alcohol and smoking, and sexual activity, were investigated in the same population. Using auto- and cross-correlation techniques, CC-potentials were characterized with parameters amplitude (A), duration (D), dominant frequency (DF), maximum cross correlation coefficient of longitudinal and bilateral CC-potential pairs (Rmax lon. and Rmax-bi.), and propagation velocity (PV ). MAIN OUTCOME MEASURES: Comparison of CC-EMG parameters A, D, DF, Rmax-lon., and Rmax-bi. between three recording sessions and assessment of the impact of confounding factors on these parameters. RESULTS: DF, D, A, and Rmax-lon. showed significant correlations among three sessions; PV showed significant correlations between two sessions performed within the same day but not between those performed on different days; Rmax-bi. did not show significant correlations between any two sessions. Intake of caffeine, alcohol, and smoking did not affect CC-potentials, while the recordings shortly after ejaculation showed more irregular oscillations and less CC-potentials with smaller A. CONCLUSIONS: CC-potential parameters DF, D, A, and Rmax-lon. have been demonstrated to be reproducible. The results provide a basis for the clinical and scientific application of CC-EMG. CC-potentials are not sensitive to confounding factors such as intake of caffeine, alcohol, and smoking, while measurements shortly after ejaculation should be avoided. PMID- 17081219 TI - Defining the clinical characteristics of Peyronie's disease in young men. AB - INTRODUCTION: Peyronie's disease (PD) is usually seen in men in their fifth decade of life. AIM: In this study, we investigated the characteristics of the disease in young men. MAIN OUTCOME MEASURES: The demographics, clinical features, and associated comorbidities of the patients with PD were retrospectively reviewed. METHODS: The findings were compared between men with the disease who were under 40 years of age with those over 40 years. Statistical analyses were conducted to define differentiating features between these two groups. RESULTS: Of the 296 patients, 32 were under the age of 40 years and 264 over 40 years. The mean duration of the disease was 2 +/- 4 and 6 +/- 8 months in the respective age groups. Fifty-six percent of the patients under the age of 40 years and 75% of the patients over this age presented with curvature (P < 0.01). Thirty-seven percent under 40 years and 12% men over 40 years had more than one plaque at presentation (P < 0.01). Dupuytren's contracture was seen only in patients over 40 years of age. Pain at presentation was found in 75% under the age of 40 years and in 65% over 40 years (P = 0.03). Trauma history was found in 18% under 40 years and in 5% over this age (P < 0.01). Statistical significant differences were found between the groups under and over the age of 40 years for hypertension (P < 0.01) and dyslipidemia (P < 0.01). Diabetes was noted in 50% of the patients under the age of 40 years and in 18% of the patients over this age (P < 0.001). Multivariate analysis of conditions associated in men with PD under 40 years of age showed statistical significant differences for diabetes (P = 0.015), presentation within 6 months (P = 0.004), and having multiple plaques (P = 0.008). CONCLUSIONS: Young men with PD are more likely to present at an earlier stage of the disease, to have diabetes, and to have more than one plaque at the time of presentation. PMID- 17081220 TI - Pilot study of sexual dysfunction following abdominal aortic aneurysm surgery. AB - INTRODUCTION: The complication of sexual dysfunction as a quality of life (QoL) component after abdominal aortic aneurysm (AAA) surgery in men is poorly studied. AIMS: To investigate the prevalence of sexual dysfunction and to highlight the importance of discussing this issue with patients undergoing AAA repair. MAIN OUTCOME MEASURES: The self-reported sexual dysfunction prevalence pre- and postoperatively, the effects on sexual QoL, and the postoperative Sexual Health Inventory for Men (SHIM) scores. METHODS: Between April 1999 and July 2002, a questionnaire-based study, including the SHIM, was conducted on male patients 1-2 years after their elective open (EO) and rupture open (RO) or endovascular repair (EVAR) AAA repair. Demographics, risk factors for sexual dysfunction, sexual history, and postoperative sexual QoL data were obtained. RESULTS: Out of 142 alive male patients surveyed, 56 (40%) patients responded (26 EO, 21 EVAR, and 9 RO repair). The mean age was 69, 73, and 70 years, respectively, and 65%, 66%, and 66%, respectively, admitted to be sexually active postoperatively. The self reported sexual dysfunction prevalence preoperatively was 27% (EO), 63% (EVAR), and 45% (RO); and postoperatively was 58%, 76%, and 67%, respectively. Detection using SHIM was higher at 70%, 95%, and 78%, respectively. There was a significantly greater increase in the postoperative prevalence of sexual dysfunction in the EO group than in the EVAR group (P < 0.05, chi(2)). The sexual QoL was worsened postoperatively in all groups: 53% (EO), 75% (EVAR), and 50% (RO); but only one-third of EO and EVAR patients, and none in RO patients, would seek treatment for their sexual dysfunction. CONCLUSION: There was a negative impact on the sexual QoL in all groups after surgery, and a significantly higher proportion of patients experienced deterioration in sexual QoL following EO surgical repair. Our results demonstrate the need for a prospective study. PMID- 17081221 TI - Association of risk factors and medical comorbidities with male sexual dysfunctions. AB - INTRODUCTION: Conventionally, little attention has been given to the association of risk factors and medical comorbidities with male sexual dysfunctions. Although that association has been recently shown in many studies, it is not yet well investigated in the Saudi community. AIM: To investigate the association of risk factors and medical comorbidities with male sexual dysfunctions in the Saudi community. METHODS: A total of 1,464 male patients with a clinical diagnosis of sexual dysfunctions were enrolled in this study. All patients were assessed for sexual functions using different domains of the International Index for Erectile Function. Patients were also interviewed for sociodemographic data, medical history, and risk factors for erectile dysfunction (ED). Routine laboratory investigations, plus total testosterone and prolactin assessments, were offered to all patients. Assessments of penile vasculature using Doppler ultrasonography and rigidometer were performed. RESULTS: A total of 92.6% of the patients had ED, 50.8% had premature ejaculation (PE), and 7.6% had low sexual desire. There was a significant association between increased age and increased severity of ED. In total, 20% had psychogenic cause, whereas 80% had organic cause of ED. Of the patients, 10.2% had mild, 41% had moderate, and 48.8% had severe ED. There were significant associations between endocrinopathy and both low sexual desire and PE (P < 0.05). There were significant associations between increased severity of ED and presence of diabetes, hypertension, dyslipidemia, ischemic heart disease, myocardial infarction, and psychological disorders. There were significant associations between increased severity of ED and increased values of end diastolic velocity, decreased values of peak systolic velocity, resistive index, rigidometer, and decreased response to intracavernosal injection (P < 0.001). CONCLUSION: This study provides an assessment of the association of risk factors and medical comorbidities with male sexual dysfunctions in ambulatory service in this community. PMID- 17081222 TI - Sexual function and obstructive sleep apnea-hypopnea: a randomized clinical trial evaluating the effects of oral-appliance and continuous positive airway pressure therapy. AB - INTRODUCTION: The obstructive sleep apnea-hypopnea syndrome (OSAHS) is associated with sexual dysfunction. Although successful treatment with continuous positive airway pressure (CPAP) has been demonstrated to improve sexual function, the effects of oral-appliance therapy are unknown. AIM: The aims of this study were to determine to what extent untreated male OSAHS patients experience sexual dysfunctions compared with control subjects, and second, to evaluate the effects of oral-appliance and CPAP therapy on sexual functioning. METHODS: Sexual functioning was determined in 48 OSAHS patients with the Golombok Rust inventory of sexual satisfaction (GRISS) and a testosterone measurement. GRISS outcomes were compared with 48 age-matched male controls without any sexual problems. Patients were randomized for either oral-appliance or CPAP therapy. After 2-3 months of treatment, the GRISS and testosterone measurements were repeated. MAIN OUTCOME MEASURE: The outcomes on the GRISS were used as the main outcome measure. RESULTS: Compared with controls, OSAHS patients had significantly more erectile dysfunction (mean +/- standard deviation; OSAHS 8.7 +/- 3.8 vs. controls 6.8 +/- 2.6) and sexual dissatisfaction (mean +/- standard deviation; OSAHS 9.7 +/- 4.2 vs. controls 8.1 +/- 2.6) as indicated by the GRISS. No significant changes in the GRISS or testosterone levels were observed in the 20 and 27 patients completing the follow-up review for oral-appliance and CPAP therapy. A correlation was demonstrated between the extent of erectile dysfunction at baseline and improvements in erectile function following treatment (r = -0.547, P = 0.000). CONCLUSIONS: This study confirms that male OSAHS patients show more sexual dysfunctions compared with age-matched control subjects. Although significant improvements in sexual functioning in neither the oral-appliance nor CPAP-treated group could be established, our findings suggest that untreated OSAHS patients with pronounced erectile dysfunction experience some improvement following treatment. PMID- 17081223 TI - Prevalence and correlates of erectile dysfunction by race and ethnicity among men aged 40 or older in the United States: from the male attitudes regarding sexual health survey. AB - INTRODUCTION: Most U.S. population-based estimates of erectile dysfunction (ED) prevalence restricted upper age, were not nationally representative, or underrepresented minority groups. AIM: To estimate, by race/ethnicity in the United States, the prevalence of ED and the impact of sociodemographic, health, relationship, psychological, and lifestyle variables. METHODS: This cross sectional, population-based, nationally representative probability survey conducted between May 2001 and January 2002 in the general community setting facilitated equivalent representation among U.S. non-Hispanic white (N = 901), non-Hispanic black (N = 596), and Hispanic (N = 676) men aged 40 and older by using targeted phone lists to oversample the minority populations. MAIN OUTCOME MEASURE: Estimated prevalence of moderate or severe ED, defined as a response of "sometimes" or "never" to the question "How would you describe your ability to get and keep an erection adequate for satisfactory intercourse?" RESULTS: The estimated prevalence was 22.0% (95% confidence interval [CI], 19.4-24.6) overall, 21.9% (95% CI, 18.8-24.9) in whites, 24.4% (95% CI, 18.4-30.5) in blacks, and 19.9% (95% CI, 13.9-25.9) in Hispanics, and increased with increasing age. The odds ratio increased with increasing age. Probability also increased with diabetes, hypertension, and moderate or severe lower urinary tract symptoms (LUTS) overall; age > or =70 years and diabetes in whites; severe LUTS in blacks; and age > or =60 years, moderate LUTS, hypertension, and depression in Hispanics. It decreased with exercise and college vs. less than high school education overall; with exercise, good relationship quality, and according to alcohol intake in blacks; and with high school or college education in Hispanics. CONCLUSIONS: The odds of ED increased with increasing age across race/ethnicity when controlling for sociodemographic, health, relationship, psychological, and lifestyle variables. These initial analyses suggest further study of the interrelationships among risk factors for ED. PMID- 17081224 TI - Perioperative fluid management in renal transplantation: a narrative review of the literature. AB - Adequate volume maintenance is essential to prevent acute renal failure during major surgery or to ensure graft function after renal transplantation. The various recommendations on the optimum fluid therapy are based, at best, on sparse evidence only from observational studies. This article reviews the literature on perioperative fluid management in renal transplantation. Crystalloid solutions not exerting any specific side-effects are the first choice for volume replacement in kidney transplantation. The use of colloids should be restricted to patients with severe intravascular volume deficits necessitating high volume restoration. The routine application of albumin, dopamine, and high dose diuretics is no longer warranted. Mannitol given immediately before removal of the vessel clamps reduces the requirement of post-transplant dialysis, but has no effects on graft function in the long term. There is insufficient evidence on the best use of dialysis, but it seems peritoneal dialysis pretransplant is associated with less delayed graft function, whereas the preference of dialysis post-transplant is not yet well-founded. This review article should provide better guidance for fluid management in kidney transplantation until best evidence guidelines can be established based upon more research. PMID- 17081225 TI - Polyomavirus nephropathy in native kidneys and renal allografts: an update on an escalating threat. AB - Polyomavirus nephropathy, also termed BK-virus nephropathy (BKN) after the main causative agent, the polyoma-BK-virus strain, is a significant complication after kidney transplantation. BKN is the most common viral infection that affects renal allografts with a prevalence of 1-9% on average 8-13 months post surgery. It can also occur sporadically in native kidneys. Viral nephropathy is caused by the (re)activation of latent BK viruses that enter into a replicative cycle under sustained and intensive immunosuppression. Pure productive kidney infections with JC- and SV-40 polyomaviruses are exceptionally rare. BKN is morphologically defined by the presence of intranuclear viral inclusion bodies in epithelial cells and tubular injury, which is the morphological correlate for renal dysfunction. Renal disease can progress through different histologic stages (from early BKN stage A to late fibrotic stage C) that carry prognostic significance; disease stages B and C often result in chronic kidney (allograft) dysfunction and end-stage renal disease. The clinical goal is to diagnose viral nephropathy in disease stage A and to limit chronic renal injury. Strategies to recognize, classify, and manage BKN are critically discussed including ancillary techniques for risk assessment and patient monitoring: (i) urine cytology and the search for so-called 'decoy cells'; (ii) PCR analyses for viral load measurements in the plasma and urine; and (iii) negative staining urine electron microscopy to identify viral particles. PMID- 17081226 TI - Vascular resistance and endothelial function in cyclosporine-treated lung transplant recipients. AB - The majority of patients undergoing solid organ transplantation develop hypertension, to which vasoconstriction and impaired endothelial function have been suggested to contribute. We compared basal vascular resistance and nitric oxide-mediated endothelial-dependent and independent vasoreactivity between cyclosporine-treated lung transplant recipients and healthy subjects. Forearm blood flow was measured by venous occlusion plethysmography at rest and during acetylcholine, glyceryltrinitrate and N(G)-monomethyl-L-arginine acetate (L-NMMA) infusion in 11 lung transplant recipients 3-5 years after transplantation and in eight healthy subjects. Forearm vascular resistance (FVR) was calculated. Plasma levels of endothelin-1 (ET-1) and von Willebrand factor (vWf) were analysed. Basal vascular resistance was 40% lower in transplant recipients than in healthy subjects (P = 0.021). Endothelial-dependent and independent vasodilation did not differ. Plasma levels of ET-1 and vWf were higher in transplant recipients (P = 0.009 and P < 0.001 respectively). There was a significant correlation between ET 1 levels and FVR in healthy subjects (r = 0.83, P = 0.042), but not in transplant recipients (r = -0.14, P = 0.70). The findings oppose the theory of generalized vasoconstriction and impaired endothelial function in the pathogenesis of hypertension after transplantation. Increased plasma levels of ET-1 do not cause increased FVR in lung transplant recipients. PMID- 17081227 TI - Systematic grading of surgical complications in live liver donors according to Clavien's system. AB - The lack of consensus on how to evaluate surgical complications of donors in live donor liver transplantation (LDLT) and incoherence of cumulative data hampers efficient comparison of the outcome worldwide. We considered that the application of the internationally validated classification system introduced by Clavien in 2004 might be beneficial. Operative complications of 243 patients who underwent live donor hepatectomy for adult LDLT between January 1996 and October 2005 at the University of Tokyo were analyzed according to the system. Definitions for each grade in the system are: grade I, deviation from the normal postoperative course but without the need for therapy; grade II, complication requiring pharmacologic treatment; grade III, complication with the need for surgical, endoscopic or radiological intervention (IIIa/b: without/with the need for general anesthesia); grade IV, life-threatening complication requiring intensive care; grade V, death. Surgical morbidity was recognized in 67 donors (28%). No deaths occurred. The numbers of patients with complications were: grade I, 36 (15%); II, 10 (4%); IIIa, 12 (5%); IIIb, 9 (4%); IV, 0; V, 0. Six in IIIb underwent surgical repair for bile leakage. Clavien's system is simple and informative. It may serve as a common tool for the quality assessment in live liver donor surgery worldwide, and we propose its application whenever surgical complication of live donor is discussed. PMID- 17081228 TI - Change in model for end-stage liver disease score on the transplant waiting list predicts survival in patients undergoing liver transplantation. AB - Allocation of donor livers through the model for end-stage liver disease (MELD) score has resulted in a fall in waiting list deaths in the United States. Change in MELD score (DeltaMELD) whilst awaiting transplant has been suggested as a method of refining organ allocation. Our aims were to analyse the effect of DeltaMELD between listing and transplant, and examine its impact on patient survival, intensive care stay and hospital stay in 402 patients transplanted for chronic liver disease at a single centre. Patients who had a DeltaMELD score of >+1 point were more likely to die in hospital following transplant (P < 0.05) and had a significantly worse 12- and 36-month survival post transplant (P < 0.0001) when compared with patients with DeltaMELD O recipient) and the other pair is compatible, but not identical (e.g. O-donor > A recipient). Exchanging these kidneys would result in two compatible living donor kidney transplants. We studied whether compatible pairs would be willing to participate in such procedure. We included 96 living kidney donors and recipients in our study. These donors and recipients could be divided into two groups: (i) donors and their direct recipients (n = 48), and (ii) paired exchange donors and their intended recipients (n = 48). All were asked whether they would be willing to participate in altruistically unbalanced exchange donation, as long as direct donation was also an option. We found no group differences. We found that one third of the donors and recipients are willing to participate in altruistically unbalanced kidney exchanges. Therefore this form of donation may be a feasible addition to already existing living kidney exchange programs. PMID- 17081230 TI - Orlistat treatment is safe in overweight and obese liver transplant recipients: a prospective, open label trial. AB - Obesity is a frequent complication following liver transplantation and is insufficiently responsive to dietary and life style advice. We studied the safety of orlistat treatment in obese and overweight liver transplant recipients (n = 15) on a stable tacrolimus-based immunosuppressive regimen. For safety reasons, the treatment period was restricted (6 months 120 mg t.i.d., 3 months 120 mg daily). Three patients dropped out, tacrolimus dose was adjusted in six of 12 remaining patients (dose reduction in 4, increase in 2, P = N.S.). All dose adjustments occurred during the 6 months of orlistat 120 mg t.i.d. therapy. No drug intolerance, adverse events or episodes of rejection occurred during the study. Efficacy of orlistat treatment in this population could not be shown, because a formal control population was not included in this safety trial. Moreover, only a significant decrease of waist circumference (P < 0.01 versus start of the study), but not of weight or body mass index, was achieved in the treated group. Orlistat treatment is well tolerated in liver transplant recipients and can be started safely, provided immunosuppressive drug levels and dietary adherence are closely monitored. PMID- 17081231 TI - Pulmonary Lophomonas blattarum infection in patients with kidney allograft transplantation. AB - The aim of the study was to analyse the clinical manifestation and management of pulmonary Lophomonas blattarum infection in four allograft transplantation recipients retrospectively. Four patients with pulmonary L. blattarum infection were diagnosed by using Fiberoptic bronchoscopy (FOB) and bronchoalveolar lavage (BAL) examination. Their clinical manifestation and management are summarized. Four cases of pulmonary L. blattarum were found during the period from the second month to the third month after transplantation. Concurring infection by other pathogens was found in three of them. Common initial symptoms included fever (>38 degrees C) without cough and breathlessness. Lower lobe shadowing could be found on chest X-ray. Body temperature decreased to the normal range in three patients and to 37.5 degrees C in the other one, after intravenous injection of metronidazole and tapering immunosuppressant. Radiological examination confirmed improved health condition of the patients afterwards. Two patients received repeated FOB and only dead L. blattarum was found. Pulmonary L. blattarum infection in allograft transplant recipients carry relatively obscure initial symptoms. Possible L. blattarum infection needs to be screened in post transplantation pulmonary infection patients with similar symptoms, especially in those who respond poorly to anti-infection treatment. Microscopic examination of BAL fluid can help to identify pulmonary L. blattarum infection and metronidazole is an ideal treatment choice. PMID- 17081232 TI - Janus kinase 3 inhibition with CP-690,550 prevents allograft vasculopathy. AB - Janus kinase 3 (JAK3) mediates signal transduction from cytokine receptors using the common gamma chain. The rationally designed inhibitor of JAK3, CP-690,550, prevents acute allograft rejection in rodents and in nonhuman primates. Here we investigated the ability of CP-690,550, to prevent allograft vasculopathy in a rodent model of aorta transplantation. Aortas from AxC Irish (RT1(a)) or Lewis (RT1(l)) rats were heterotopically transplanted into the infra-renal aorta of Lewis recipients and harvested at 28 or 56 days. Treated recipients received CP 690,550 by osmotic pumps (mean drug exposure of 110 +/- 38 ng/ml). Significant intimal hyperplasia was demonstrated in untreated allografts when compared with isografts at 28 days (2.08 +/- 0.85% vs. 0.43 +/- 0.2% luminal obliteration, respectively, P = 0.001) and 56 days (5.3 +/- 2.4% vs. 0.38 +/- 0.3%, P = 0.002). Treatment caused a 51% reduction in intimal hyperplasia at day 56. CP-690,550 treated animals also had a significant reduction of donor-specific IgG production and of the gene expression for suppressor of cytokine signaling-3 and with unchanged levels of expression of RANTES, IP-10 and transforming growth factor beta1. These results are the first to show that JAK3 blockade by CP-690,550 effectively prevents allograft vasculopathy in this rat model of aorta transplantation. PMID- 17081233 TI - Sirolimus as the main immunosuppressant in the early postoperative period following liver transplantation: a report of six cases and review of the literature. AB - The use of sirolimus as the main immunosuppressant in a calcineurin inhibitor free regimen in the early postoperative period of liver transplantation (LT), when the incidence of rejection is the highest, has seldom been reported. We report six patients who received sirolimus in association with steroids only, at a median time of 10 days after LT (range 3-23). Tacrolimus, initially given as the standard immunosuppressant, was discontinued because of nephrotoxicity in three of these patients and neurotoxicity in the other three. Resolution of the neurological symptoms was observed in all cases and a marked improvement of the renal function in two of three patients. Two patients died, one of sepsis and the other of recurrent hepatitis C virus hepatitis, after 47 and 143 days respectively. Three patients developed acute rejection which responded to intravenous steroids. In this cohort of patients, the use of sirolimus appeared safe and provided an adequate prophylaxis against rejection, even though the drug was administered in the immediate postoperative period after LT. PMID- 17081234 TI - Cholestasis in pregnancy associated with ciclosporin therapy in renal transplant recipients. AB - Obstetric cholestasis (OC) presents with pruritus in the second half of pregnancy and is associated with increased risk of foetal distress, intra-uterine death and premature delivery. From a tertiary referral, renal-obstetric clinic, we report the occurrence of OC in 5/23 pregnancies of women with renal transplants maintained on ciclosporin treatment (European incidence 0.1-1.5% of pregnancies). All required premature delivery for foetal reasons at 33-37/40 (median 34/40). Ciclosporin, at therapeutic concentrations, inhibits bile salt excretion pump (BSEP) function in rats and humans. We propose that OC developed in our patients because the mild inhibition of the canalicular pumps by ciclosporin was only revealed in pregnancy when increases in progesterone metabolites overwhelmed pump function. We suggest that all pregnant women receiving ciclosporin should be closely monitored from the second trimester for the development of OC. If detected, enhanced foetal and maternal monitoring to optimize time of delivery and pregnancy outcome is required. PMID- 17081235 TI - Liver transplantation of a HELLP syndrome donor liver in acute fulminant hepatitis B with deltavirus coinfection. PMID- 17081236 TI - Temporary transjugular intrahepatic portosystemic shunt to overcome small-for size syndrome after right lobe adult split liver transplantation. PMID- 17081237 TI - Allelochemical stress causes inhibition of growth and oxidative damage in Lycopersicon esculentum Mill. AB - The aim of this study was to analyse the effect of allelochemical stress on Lycopersicon esculentum growth. Our results showed that allelochemical stress caused by Sicyos deppei aqueous leachate inhibited root growth but not germination, and produced an imbalance in the oxidative status of cells in both ungerminated seeds and in primary roots. We observed changes in activity of catalase (CAT), ascorbate peroxidase (APX), superoxide dismutase (SOD), glutathione reductase (GR) and the plasma membrane NADPH oxidase, as well as in the levels of H(2)O(2) and O(2) (*-) in seeds at 12 and 24 h, and in primary roots at 48 and 72 h of treatment, which could account for the oxidative imbalance. There were changes in levels of expression of the mentioned enzymes, but without a correlation with their respective activities. Higher levels of membrane lipid peroxidation were observed in primary roots at 48 and 72 h of treatment. No effect on the expression of metacaspase and the PR1 was observed as indicators of cell death or induction of plant defence. This paper contributes to the understanding of plant-plant interactions through the phytotoxic allelochemicals released in an aqueous leachate of the weed S. deppei, which cause a negative effect on other plants. PMID- 17081238 TI - Cyclitols and carbohydrates in leaves and roots of 13 Eucalyptus species suggest contrasting physiological responses to water deficit. AB - In many tree species, physiological adaptations to drought include the accumulation of osmotically active substances and/or the presence of particular compatible solutes, among them cyclitols. Recently, the cyclitol quercitol was identified in species of Eucalyptus, a diverse genus whose speciation is probably driven by adaptation to water availability. We subjected seedlings of 13 Eucalyptus species from different ecosystems ('mesic' and 'xeric') and different sub-generic taxonomic groups to 10 weeks of water deficit (WD) treatment. Pre dawn water potentials (psi(pdwn)) and relative water content (RWC) were determined in shoots, and total osmolality, soluble low-molecular-weight carbohydrates and cyclitols were measured in leaves and roots. Responses to water deficit followed two distinct patterns: Eucalyptus species from 'mesic' environments adjusted concentrations of sucrose (through increased levels of sucrose and decreases in RWC) in response to water deficit, whereas 'xeric' species increased concentrations of quercitol (through reductions in RWC). In root tissues, only species from xeric environments contained high levels of quercitol and mannitol, increasing under WD conditions. We suggest that the former (mesic) strategy may be beneficial to respond to short-lasting drought conditions, because sucrose is easily metabolized, whereas the latter (xeric) strategy may relate to an effective acclimation to longer-lasting drought. These physiological response groups are also related to taxonomic groups within the genus. PMID- 17081239 TI - Biomass partitioning and leaf N,P - stoichiometry: comparisons between tree and herbaceous current-year shoots. AB - We compare the biomass partitioning patterns and the nitrogen/phosphorus (N,P) stoichiometry of the current-year shoots of tree and herbaceous species and ask whether they scale in the same ways. Our analyses indicate that few statistically significant differences exist between the shoot biomass partitioning patterns of the two functional species-groups. In contrast, statistically significant N,P - stoichiometric differences exist between the two functional groups. Across all species, dry leaf mass scales nearly as the square of basal stem diameter and isometrically with respect to dry stem mass. However, total leaf N scales as the 1.37-power and as the 1.09-power of total leaf P across herbaceous and tree shoots, respectively. Therefore, tree shoots can be viewed as populations of herbs elevated by their older, woody herbaceous cohorts. However, tree leaf stoichiometry cannot be modelled in terms of herbaceous N,P - leaf stoichiometry. PMID- 17081240 TI - Parallel analysis of transcript levels and physiological key parameters allows the identification of stress phase gene markers in Chlamydomonas reinhardtii under copper excess. AB - Excessive copper concentrations, known to induce reactive oxygen species (ROS) formation, have been tested with respect to their effects on transcript abundance and related proteins involved in oxidative stress responses. The results show that the stromal photosynthetic functions were more ROS sensitive than the membrane-located reactions. The rbcL over-expression compensated for the damage only at 10 microM Cu, whereas the genetic stimulation of alpha-tocopherol biosynthesis led to the protection of membrane reactions up to 50-100 microM Cu. For this reason, the gradual growth drop of Chlamydomonas reinhardtii cultures observed under increasing Cu(2+) concentrations matched better with the loss of photosynthetic capacity than with those of photosynthetic quantum yields. According to Larcher's stress concept, the results allow the identification of gene markers for the alarm (rbcL), the hardening (FeSOD, VTE3) and the exhaustion [cyclin-dependent protein kinase (cdk), psbA] phases. These genes can be used to rapidly evaluate the state of oxidative stress in algae and putatively in other plant cells. PMID- 17081241 TI - Impact of the C-N status on the amino acid profile in tobacco source leaves. AB - This paper investigates the influence of the carbon (C) and nitrogen (N) status on the amino acid profile in tobacco source leaves. Treatments used included growing plants at different light intensities, using an antisense RBCS (small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase) construct to inhibit Rubisco activity, growing plants on 12 or 0.5 mM nitrate, comparing wild-types with genotypes that have small and large decreases in nitrate reductase (NIA) activity, and sampling plants at different times during the diurnal cycle. This combination of experiments provides information on how amino acid levels respond to several inputs including the C and N status, nitrate, excess light and light dark transitions. The data set was analysed using principal component analysis, regression analysis and by normalizing the level of each individual amino acid on the total amino acid pool. Most amino acids show a downward trend when the C or the N status is decreased, and rise during day and fall at night during the diurnal cycle. However, individual amino acids often showed deviating responses. Furthermore, no evidence was found for feedback inhibition of minor amino acid synthesis, either within or between pathways, when 18 individual amino acids were supplied to detached leaves. Results indicate that regulation of amino acid metabolism, for example by the C and N status, leads to qualitatively similar responses of many amino acids, but homeostatic mechanisms involving feedback inhibition within or between individual amino acid biosynthesis pathways are not stringent. All of the above inputs affect the level of phenylalanine, an amino acid that is also the substrate for an important sector of secondary metabolism. The levels of glutamate were remarkably constant, indicating that unknown mechanisms stabilize the concentration of this key central amino acid. Analyses of metabolite levels and feeding experiments indicated that 2-oxoglutarate plays an important role in regulating glutamate levels. Glutamate was the most effective inhibitor of NIA activity when 18 individual amino acids were supplied to detached leaves. Feeding glutamate, and other downstream amino acids, led to an increase of glutamine, indicating glutamate exerts feedback regulation on ammonium metabolism. PMID- 17081242 TI - Hourly and seasonal variation in photosynthesis and stomatal conductance of soybean grown at future CO(2) and ozone concentrations for 3 years under fully open-air field conditions. AB - It is anticipated that enrichment of the atmosphere with CO(2) will increase photosynthetic carbon assimilation in C3 plants. Analysis of controlled environment studies conducted to date indicates that plant growth at concentrations of carbon dioxide ([CO(2)]) anticipated for 2050 ( approximately 550 micromol mol(-1)) will stimulate leaf photosynthetic carbon assimilation (A) by 20 to 40%. Simultaneously, concentrations of tropospheric ozone ([O(3)]) are expected to increase by 2050, and growth in controlled environments at elevated [O(3)] significantly reduces A. However, the simultaneous effects of both increases on a major crop under open-air conditions have never been tested. Over three consecutive growing seasons > 4700 individual measurements of A, photosynthetic electron transport (J(PSII)) and stomatal conductance (g(s)) were measured on Glycine max (L.) Merr. (soybean). Experimental treatments used free air gas concentration enrichment (FACE) technology in a fully replicated, factorial complete block design. The mean A in the control plots was 14.5 micromol m(-2) s(-1). At elevated [CO(2)], mean A was 24% higher and the treatment effect was statistically significant on 80% of days. There was a strong positive correlation between daytime maximum temperatures and mean daily integrated A at elevated [CO(2)], which accounted for much of the variation in CO(2) effect among days. The effect of elevated [CO(2)] on photosynthesis also tended to be greater under water stress conditions. The elevated [O(3)] treatment had no statistically significant effect on mean A, g(s) or J(PSII) on newly expanded leaves. Combined elevation of [CO(2)] and [O(3)] resulted in a slightly smaller increase in average A than when [CO(2)] alone was elevated, and was significantly greater than the control on 67% of days. Thus, the change in atmospheric composition predicted for the middle of this century will, based on the results of a 3 year open-air field experiment, have smaller effects on photosynthesis, g(s) and whole chain electron transport through photosystem II than predicted by the substantial literature on relevant controlled environment studies on soybean and likely most other C3 plants. PMID- 17081243 TI - VfCPK1, a gene encoding calcium-dependent protein kinase from Vicia faba, is induced by drought and abscisic acid. AB - Calcium, one of the most ubiquitous second messengers, has been shown to be involved in a wide variety of responses in plants. Calcium-dependent protein kinases (CDPKs) (EC 2.7.1.37) are the predominant Ca(2+)-regulated serine/threonine protein kinase in plants and play an important role in plant calcium signal transduction. CDPKs are encoded by a large multigene family in many plants, which has been showed so far; however, the precise role of each specific CDPK is still largely unknown. A novel CDPK gene designated as VfCPK1 was cloned from epidermal peels of broad bean (Vicia faba L.) leaves using the rapid amplification of cDNA ends (RACE)-PCR technique and its expression was studied in detail. The VfCPK1 cDNA is 1783 bp long and contains an open reading frame of 1482 bp encoding 493 amino acids. VfCPK1 contains all conserved regions found in CDPKs and shows a high level of sequence similarity to many other plant CDPKs. VfCPK1 was highly expressed in leaves, especially in leaf epidermal peels of broad bean in mRNA and protein levels. Expressions of VfCPK1 at both the mRNA and protein levels were increased in leaves treated with abscisic acid or subjected to drought stress. Potential roles of VfCPK1 in epidermal peels are discussed. The nucleotide sequence data reported here were deposited in the GenBank database under accession number AY753552. PMID- 17081244 TI - Changes in starch and inositol 1,4,5-trisphosphate levels and auxin transport are interrelated in graviresponding oat (Avena sativa) shoots. AB - This study was conducted to unravel a mechanism for the gravitropic curvature response in oat (Avena sativa) shoot pulvini. For this purpose, we examined the downward movement of starch-filled chloroplast gravisensors, differential changes in inositol 1,4,5-trisphosphate (IP(3)) levels, transport of indole-3-acetic acid (IAA) and gravitropic curvature. Upon gravistimulation, the ratio for IAA levels in lower halves versus those in upper halves (L/U) increased from 1.0 at 0 h and reached a maximum value of 1.45 at 8 h. When shoots were grown in the dark for 10 d, to deplete starch in the chloroplast, the gravity-induced L/U of IAA was reduced to 1.0. N-naphthylphthalamic acid (NPA) and 2,3,5-triiodobenzoic acid (TIBA), both auxin transport inhibitors, significantly reduced the amount of gravitropic curvature and gravity-induced lateral IAA transport, but did not reduce the gravity-induced late change in the L/U ratio of IP(3) levels. U73122, a specific phospholipase C (PLC) inhibitor, decreased gravity-induced curvature. Because U73122 reduced the ratio of L/U of IAA imposed by gravistimulation, it is clear that IAA transport is correlated with changes in IP(3) levels upon gravistimulation. These results indicate that gravistimulation-induced differential lateral IAA transport may result from the onset of graviperception in the chloroplast gravisensors coupled with gravity-induced asymmetric changes in IP(3) levels in oat shoot pulvini. PMID- 17081245 TI - Phosphoenolpyruvate carboxylase genes in C3, crassulacean acid metabolism (CAM) and C3/CAM intermediate species of the genus Clusia: rapid reversible C3/CAM switches are based on the C3 housekeeping gene. AB - The genus Clusia includes species that exhibit either the C3 or crassulacean acid metabolism (CAM) mode of photosynthesis, or those that are able to switch between both modes according to water availability. In order to screen for species specific genetic variability, we investigated the key carboxylase for CAM, phosphoenolpyruvate carboxylase (PEPC). Sequence analysis of DNA isolated from the obligate CAM species, Clusia hilariana, the obligate C3 species, Clusia multiflora, and an intermediate species that can switch between C3 and CAM photosynthesis, Clusia minor, revealed three different isoforms for C. hilariana and one each for the other two species. Sequence alignments indicated that PEPC from the intermediate species had high homology with the C3 protein and with one of CAM plant proteins. These were assumed to constitute 'housekeeping' proteins, which can also support CAM in intermediate species. The other two isoforms of the CAM plant C. hilariana were either CAM-specific or showed homologies with PEPC from roots. Phylogenetic trees derived from neighbour-joining analysis of amino acid sequences from 13 different Clusia species resulted in two distinct groups of plants with either 'housekeeping' PEPC only, or additionally CAM-related isoforms. Only C. hilariana showed the third, probably root-specific isoform. The high homology of the PEPC from the intermediate species with the C3 protein indicates that for the reversible transition from the C3 to CAM mode of photosynthesis, the C3 type of PEPC is sufficient. Its expression, however, is strongly increased under CAM-inducing conditions. The use of the C3 isoform could have facilitated the evolution of CAM within the genus, which occurred independently for several times. PMID- 17081246 TI - Al-Fe interactions and growth enhancement in Melastoma malabathricum and Miscanthus sinensis dominating acid sulphate soils. AB - Plants growing in acid sulphate soils are subject to high levels of Al availability, which may have effects on the growth and distribution of these species. Although Fe availability is also high in acid sulphate soils, little is known about the effect of Fe on the growth of native plants in these soils. Two species dominating this soil type in Asia, viz. Melastoma malabathricum and Miscanthus sinensis were grown hydroponically in a nutrient solution with different concentrations of Al and Fe. Melastoma malabathricum is found to be sensitive to Fe (40 and 100 microm). Application of 500 microm Al, however, completely ameliorates Fe toxicity and is associated with a decrease of Fe concentration in shoots and roots. The primary reason for the Al-induced growth enhancement of M. malabathricum is considered to be the Al-induced reduction of toxic Fe accumulation in roots and shoots. Therefore, Al is nearly essential for M. malabathricum when growing in acid sulphate soils. In contrast, application of both Fe and Al does not reduce the growth of M. sinensis, and Al application does not result in lower shoot concentrations of Fe, suggesting that this grass species has developed different mechanisms for adaptation to acid sulphate soils. PMID- 17081247 TI - The impact of coral bleaching on the pigment profile of the symbiotic alga, Symbiodinium. AB - Bleaching of corals by loss of symbiotic dinoflagellate algae and/or photosynthetic pigments is commonly triggered by elevated temperatures coupled with high irradiance, and is a first-order threat to coral reef communities. In this study, a high-resolution high-performance liquid chromatography method integrated with mass spectrometry was applied to obtain the first definitive identification of chlorophyll and carotenoid pigments of three clades of symbiotic dinoflagellate algae (Symbiodinium) in corals, and their response to experimentally elevated temperature and irradiance. The carotenoids peridinin, dinoxanthin, diadinoxanthin (Dn), diatoxanthin (Dt) and beta-carotene were detected, together with chlorophylls a and c2, and phaeophytin a, in all three algal clades in unstressed corals. On exposure to elevated temperature and irradiance, three coral species (Montastrea franksi and Favia fragum with clade B algae, and Montastrea cavernosa with clade C) bleached by loss of 50-80% of their algal cells, with no significant impact to chlorophyll a or c2, or peridinin in retained algal cells. One species (Agaricia sp. with clade C) showed no significant reduction in algal cells at elevated temperature and irradiance, but lost substantial amounts of chlorophyll a and carotenoid pigments, presumably through photo-oxidative processes. Two coral species (Porites astreoides and Porites porites both bearing clade A algae) did not bleach. The impact of elevated temperature and irradiance on the levels of the photoprotective xanthophylls (Dn + Dt) and beta-carotene varied among the corals, both in pool size and xanthophyll cycling, and was not correlated to coral bleaching resistance. PMID- 17081248 TI - Drought stress response in wheat: physiological and molecular analysis of resistant and sensitive genotypes. AB - Water deficit is a severe environmental stress and the major constraint on plant productivity with an evident effect on plant growth. The aim of this work was to study Triticum and Aegilops seedlings differing in their response to drought stress at the physiological and molecular levels. The identification of resistant and sensitive genotypes was firstly based on the relative water content (RWC) measurement. Further characterization of genotypes contrasting in their response to water stress was performed at the physiological level by determination of RWC, water loss rate (WLR) and free proline content after different hours of dehydration. Modification in the expression level of five dehydrin (DHN) genes was also analysed by reverse transcription-polymerase chain reaction (RT-PCR). Five cDNAs coding for different DHNs were identified and characterized. These genes are not expressed in the well-watered plants, but only in the stressed plants. Four of these cDNAs are related to novel DHN sequences. The results obtained clearly indicate a relation between the expression of these genes and tissue water content. In particular, in the resistant genotypes the expression of DHN genes is initiated even though tissue hydration levels are still high, indicating also in wheat the involvement of these proteins in water retention. PMID- 17081249 TI - Nutrient availability constrains the hydraulic architecture and water relations of savannah trees. AB - Leaf and whole plant-level functional traits were studied in five dominant woody savannah species from Central Brazil (Cerrado) to determine whether reduction of nutrient limitations in oligotrophic Cerrado soils affects carbon allocation, water relations and hydraulic architecture. Four treatments were used: control, N additions, P additions and N plus P additions. Fertilizers were applied twice yearly, from October 1998 to March 2004. Sixty-three months after the first nutrient addition, the total leaf area increment was significantly greater across all species in the N- and the N + P-fertilized plots than in the control and in the P-fertilized plots. Nitrogen fertilization significantly altered several components of hydraulic architecture: specific conductivity of terminal stems increased with N additions, whereas leaf-specific conductivity and wood density decreased in most cases. Average daily sap flow per individual was consistently higher with N and N + P additions compared to the control, but its relative increase was not as great as that of leaf area. Long-term additions of N and N + P caused midday PsiL to decline significantly by a mean of 0.6 MPa across all species because N-induced relative reductions in soil-to-leaf hydraulic conductance were greater than those of stomatal conductance and transpiration on a leaf area basis. Phosphorus-fertilized trees did not exhibit significant changes in midday PsiL. Analysis of xylem vulnerability curves indicated that N fertilized trees were significantly less vulnerable to embolism than trees in control and P-fertilized plots. Thus, N-induced decreases in midday PsiL appeared to be almost entirely compensated by increases in resistance to embolism. Leaf tissue water relations characteristics also changed as a result of N-induced declines in minimum PsiL: osmotic potential at full turgor decreased and symplastic solute content on a dry matter basis increased linearly with declining midday PsiL across species and treatments. Despite being adapted to chronic nutrient limitations, Cerrado woody species apparently have the capacity to exploit increases in nutrient availability by allocating resources to maximize carbon gain and enhance growth. The cost of increased allocation to leaf area relative to water transport capacity involved increased total water loss per plant and a decrease in minimum leaf water potentials. However, the risk of increased embolism and turgor loss was relatively low as xylem vulnerability to embolism and leaf osmotic characteristics changed in parallel with changes in plant water status induced by N fertilization. PMID- 17081250 TI - Low stomatal and internal conductance to CO2 versus Rubisco deactivation as determinants of the photosynthetic decline of ageing evergreen leaves. AB - A novel A-Ci curve (net CO2 assimilation rate of a leaf -An- as a function of its intercellular CO2 concentration -Ci) analysis method (Plant, Cell & Environment 27, 137-153, 2004) was used to estimate the CO2 transfer conductance (gi) and the maximal carboxylation (Vcmax) and electron transport (Jmax) potentials of ageing, non-senescing Pseudotsuga menziesii leaves in relation to their nitrogen (N) content and protein and pigment composition. Both gi and the stomatal conductance (gsc) of leaves were closely coupled to Vcmax, Jmax and An with all variables decreasing with increasing leaf age. Consequently, both Ci and Cc (chloroplastic CO2 concentration) remained largely conserved through successive growing seasons. The N content of leaves, as well as the amount of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and other sodium dodecyl sulfate-soluble proteins, increased during the first three growing seasons, then stabilized or decreased only slightly afterwards. Thus, the age-related photosynthetic nitrogen use efficiency (PNUE) decline of leaves was not a consequence of decreased allocation of N towards Rubisco and other proteins involved in bioenergetics and light harvesting. Rather, loss of photosynthetic capacity was the result of the decreased activation state of Rubisco and proportional down-regulation of electron transport towards the photosynthetic carbon reduction (PCR) and photorespiratory (PCO) cycles in response to a reduction of CO2 supply to the chloroplasts' stroma. This study emphasizes the regulatory potential and homeostaticity of Cc- rather than photosynthetic metabolites or Ci- in relation to the commonly observed correlation between photosynthesis and gsc. PMID- 17081251 TI - Photosynthetic capacity is related to the cellular and subcellular partitioning of Na+, K+ and Cl- in salt-affected barley and durum wheat. AB - The capacity of plants to tolerate high levels of salinity depends on the ability to exclude salt from the shoot, or to tolerate high concentrations of salt in the leaf (tissue tolerance). It is widely held that a major component of tissue tolerance is the capacity to compartmentalize salt into safe storage places such as vacuoles. This mechanism would avoid toxic effects of salt on photosynthesis and other key metabolic processes. To test this, the relationship between photosynthetic capacity and the cellular and subcellular distribution of Na+, K+ and Cl- was studied in salt-sensitive durum wheat (cv. Wollaroi) and salt tolerant barley (cv. Franklin) seedlings grown in a range of salinity treatments. Photosynthetic capacity parameters (Vcmax, Jmax) of salt-stressed Wollaroi decreased at a lower leaf Na+ concentration than in Franklin. Vacuolar concentrations of Na+, K+ and Cl- in mesophyll and epidermal cells were measured using cryo-scanning electron microscopy (SEM) X-ray microanalysis. In both species, the vacuolar Na+ concentration was similar in mesophyll and epidermal cells, whereas K+ was at higher concentrations in the mesophyll, and Cl- higher in the epidermis. The calculated cytoplasmic Na+ concentration increased to higher concentrations with increasing bulk leaf Na+ concentration in Wollaroi compared to Franklin. Vacuolar K+ concentration was lower in the epidermal cells of Franklin than Wollaroi, resulting in higher cytoplasmic K+ concentrations and a higher K+ : Na+ ratio. This study indicated that the maintenance of photosynthetic capacity (and the resulting greater salt tolerance) at higher leaf Na+ levels of barley compared to durum wheat was associated with the maintenance of higher K+, lower Na+ and the resulting higher K+ : Na+ in the cytoplasm of mesophyll cells of barley. PMID- 17081252 TI - Homeostasis of cell composition during prolonged darkness. AB - The chemical composition of organisms in relation to their environmental resource availability is an area of intense research activity. We studied the changes in cell composition of the cyanobacterium Phormidium autumnale in response to prolonged darkness. Cells allocated their internal resources in a homeostatic manner, oxidizing all the three major cellular constituents in a proportional way. This resulted in constant C/N and carbohydrates, lipids and proteins ratios that remained unaltered throughout the whole incubation period. We propose the maintenance of balanced cell composition (homeostasis) as an evolutionary strategy related to environmental transitory changes. PMID- 17081253 TI - Declining hydraulic efficiency as transpiring leaves desiccate: two types of response. AB - The conductance of transpiring leaves to liquid water (Kleaf) was measured across a range of steady-state leaf water potentials (Psileaf). Manipulating the transpiration rate in excised leaves enabled us to vary Psileaf in the range -0.1 MPa to less than -1.5 MPa while using a flowmeter to monitor the transpiration stream. Employing this technique to measure how desiccation affects Kleaf in 19 species, including lycophytes, ferns, gymnosperms and angiosperms, we found two characteristic responses. Three of the six angiosperm species sampled maintained a steady maximum Kleaf while Psileaf remained above -1.2 MPa, although desiccation of leaves beyond this point resulted in a rapid decline in Kleaf. In all other species measured, declining Psileaf led to a proportional decrease in Kleaf, such that midday Psileaf of unstressed plants in the field was sufficient to depress Kleaf by an average of 37%. It was found that maximum Kleaf was strongly correlated with maximum CO2 assimilation rate, while Kleaf = 0 occurred at a Psileaf slightly less negative than at leaf turgor loss. A strong linear correlation across species between Psileaf at turgor loss and Psileaf at Kleaf = 0 raises the possibility that declining Kleaf was related to declining cell turgor in the leaf prior to the onset of vein cavitation. The vulnerability of leaves rehydrating after desiccation was compared with vulnerability of leaves during steady-state evaporation, and differences between methods suggest that in many cases vein cavitation occurs only as Kleaf approaches zero. PMID- 17081254 TI - Plasticity to soil water deficit in Arabidopsis thaliana: dissection of leaf development into underlying growth dynamic and cellular variables reveals invisible phenotypes. AB - Genetic variability in the plasticity of leaf area expansion in response to water deficit has been reported in Arabidopsis thaliana. Here, the objective was to identify the underlying dynamic and cellular processes involved in this variability. Twenty-five accessions were subjected to identical soil water deficit treatments. In all accessions, the plasticity of leaf production was low compared with that of individual leaf expansion. A subset of accessions was selected for further dissection of individual leaf expansion into its underlying variables: the rate and duration of leaf expansion and epidermal cell number and area. In all accessions, water deficit had opposite effects on the rate and duration of leaf expansion. The accumulation of these effects was reflected in changes in final leaf area. At the cellular level, moderate water deficits had opposite effects on cell number and cell size, but more severe ones reduced both variables. The importance of these opposing effects is highlighted by the behaviour of the accession An-1, for which the compensation between the decrease in leaf expansion rate and the increase in the duration of expansion is total. This dynamic plasticity in response to water deficit is not detectable when only final measurements are done. PMID- 17081255 TI - The cytosolic Na+ : K+ ratio does not explain salinity-induced growth impairment in barley: a dual-tracer study using 42K+ and 24Na+. AB - It has long been believed that maintenance of low Na+ : K+ ratios in the cytosol of plant cells is critical to the plant's ability to tolerate salinity stress. Direct measurements of such ratios, however, have been few. Here we apply the non invasive technique of compartmental analysis, using the short-lived radiotracers 42K+ and 22Na+, in intact seedlings of barley (Hordeum vulgare L.), to evaluate unidirectional plasma membrane fluxes and cytosolic concentrations of K+ and Na+ in root tissues, under eight nutritional conditions varying in levels of salinity and K+ supply. We show that Na+ : K+ ratios in the cytosol of root cells adjust significantly across the conditions tested, and that these ratios are poor predictors of the plant's growth response to salinity. Our study further demonstrates that Na+ is subject to rapid and futile cycling at the plasma membrane at all levels of Na+ supply, independently of external K+, while K+ influx is reduced by Na+, from a similar baseline, and to a similar extent, at both low and high K+ supply. We compare our results to those of other groups, and conclude that the maintenance of the cytosolic Na+ : K+ ratio is not central to plant survival under NaCl stress. We offer alternative explanations for sodium sensitivity in relation to the primary acquisition mechanisms of Na+ and K+. PMID- 17081256 TI - Herbivory mitigation through increased water-use efficiency in a leaf-mining moth apple tree relationship. AB - Herbivory alters plant gas exchange but the effects depend on the type of leaf damage. In contrast to ectophagous insects, leaf miners, by living inside the leaf tissues, do not affect the integrity of the leaf surface. Thus, the effect of leaf miners on CO2 uptake and water-use efficiency by leaves remains unclear. We explored the impacts of the leaf-mining moth Phyllonorycter blancardella (Lepidoptera: Gracillariidae) on light responses of the apple leaf gas exchanges to determine the balance between the negative effects of reduced photosynthesis and potential positive impacts of increased water-use efficiency (WUE). Gas exchange in intact and mined leaf tissues was measured using an infrared gas analyser. The maximal assimilation rate was slightly reduced but the light response of net photosynthesis was not affected in mined leaf tissues. The transpiration rate was far more affected than the assimilation rate in the mine integument as a result of stomatal closure from moderate to high irradiance level. The WUE was about 200% higher in the mined leaf tissues than in intact leaf portions. Our results illustrate a novel mechanism by which plants might minimize losses from herbivore attacks; via trade-offs between the negative impacts on photosynthesis and the positive effects of increased WUE. PMID- 17081259 TI - Retraction: Hua, Z.-M., Yang X. & Fromm M.E. (2006). Activation of the NaCl- and drought-induced RD29A and RD29B promoters by constitutively active Arabidopsis MAPKK or MAPK proteins. Plant, Cell and Environment 29, 1761-1770. doi: 10.1111/j.1365-3040.2006.01552.x. PMID- 17081261 TI - Profiles of executive function in parents and siblings of individuals with autism spectrum disorders. AB - Delineation of a cognitive endophenotype for autism is useful both for exploring the genetic mechanisms underlying the disorder and for identifying which cognitive traits may be primary to it. This study investigated whether first degree relatives of individuals with autism spectrum disorders (ASDs) demonstrate a specific profile of performance on a range of components of executive function (EF), to determine whether EF deficits represent possible endophenotypes for autism. Parents and siblings of ASD and control probands were tested on EF tasks measuring planning, set-shifting, inhibition and generativity. ASD parents showed poorer performance than control parents on a test of ideational fluency or generativity, and ASD fathers demonstrated a weakness in set-shifting to a previously irrelevant dimension. ASD siblings revealed a mild reduction in ideational fluency and a weakness in non-verbal generativity when compared with control siblings. Neither ASD parents nor siblings displayed significant difficulties with planning or inhibition. These results indicated that the broad autism phenotype may not be characterized primarily by impairments in planning and cognitive flexibility, as had been previously proposed. Weaknesses in generativity emerged as stronger potential endophenotypes in this study, suggesting that this aspect of EF should play a central role in cognitive theories of autism. However, discrepancies in the EF profile demonstrated by parents and siblings suggest that factors related to age or parental responsibility may affect the precise pattern of deficits observed. PMID- 17081262 TI - Association between the CHRM2 gene and intelligence in a sample of 304 Dutch families. AB - The CHRM2 gene is thought to be involved in neuronal excitability, synaptic plasticity and feedback regulation of acetylcholine release and has previously been implicated in higher cognitive processing. In a sample of 667 individuals from 304 families, we genotyped three single-nucleotide polymorphisms (SNPs) in the CHRM2 gene on 7q31-35. From all individuals, standardized intelligence measures were available. Using a test of within-family association, which controls for the possible effects of population stratification, a highly significant association was found between the CHRM2 gene and intelligence. The strongest association was between rs324650 and performance IQ (PIQ), where the T allele was associated with an increase of 4.6 PIQ points. In parallel with a large family-based association, we observed an attenuated - although still significant - population-based association, illustrating that population stratification may decrease our chances of detecting allele-trait associations. Such a mechanism has been predicted earlier, and this article is one of the first to empirically show that family-based association methods are not only needed to guard against false positives, but are also invaluable in guarding against false negatives. PMID- 17081263 TI - Genetic contributions to the association between height and intelligence: Evidence from Dutch twin data from childhood to middle age. AB - A positive association between intelligence (IQ) and height has been reported previously. It is generally assumed that this association reflects the effect of childhood environment on IQ, but there is still little research supporting directly this hypothesis. We studied the association between height and IQ in 209 Dutch twin pairs at the ages of 5, 7, 10 and 12 years, 208 twin pairs at 16 and 18 years of age and 567 twin pairs and their siblings in adulthood. The heritability of height was high in all cohorts and across all ages (a2 = 0.93 - 0.96). In adulthood, heritability was also high for full-scale IQ (FSIQ: a2 = 0.83-0.84) and somewhat lower for verbal IQ (VIQ: a2 = 0.66-0.84). In early childhood, the heritability was lower, and common environmental factors had a substantial effect on FSIQ and VIQ. A positive association of height and IQ was found in early childhood and adolescence. In adulthood, a correlation was found between height and FSIQ in young adulthood and between height and VIQ in middle age. All correlations could be ascribed to genetic factors influencing both height and IQ. Thus, these results show that the association between height and IQ should not be directly regarded as evidence for childhood living conditions affecting IQ, but the effect of genetic factors affecting independently or interacting with environmental factors should be considered as well. PMID- 17081264 TI - Selective breeding for catalepsy changes the distribution of microsatellite D13Mit76 alleles linked to the 5-HT serotonin receptor gene in mice. AB - Catalepsy (pronounced motor inhibition) is a natural defensive reaction against predator. Recently, the quantitative trait locus for catalepsy was mapped on mouse chromosome 13 near the 5-HT(1A) serotonin receptor gene. Here, the linkage between catalepsy and the 5-HT(1A) receptor gene was verified using breeding experiment. Selective breeding for high predisposition to catalepsy was started from backcross BC[CBA x (CBA x AKR)] generation between catalepsy-prone (CBA) and catalepsy-resistant (AKR) mouse strains. CBA and AKR strains also differed in the 5-HT(1A) receptor functional activity. A rapid increase of cataleptic percentage from 21.2% in the backcrosses to 71% in the third generation of selective breeding (S3) was shown. The fragment of chromosome 13 including the 5-HT(1A) receptor gene was marked with D13Mit76 microsatellite. Breeding for catalepsy increased the concentration of CBA-derived and decreased the concentration of AKR derived alleles of microsatellite D13Mit76 in the S1 and S2. All mice of the S9 and S12 were homozygous for CBA-derived allele of D13Mit76 marker. Mice of the S12 showed CBA-like receptor activity. These findings indicate that selective breeding for behavior can involve selection of polymorphic variants of the 5 HT(1A) receptor gene. PMID- 17081265 TI - Thermal nociception in adult Drosophila: behavioral characterization and the role of the painless gene. AB - Nociception, warning of injury that should be avoided, serves an important protective function in animals. In this study, we show that adult Drosophila avoids noxious heat by a jump response. To quantitatively analyze this nociceptive behavior, we developed two assays. In the CO2 laser beam assay, flies exhibit this behavior when a laser beam heats their abdomens. The consistency of the jump latency in this assay meets an important criterion for a good nociceptive assay. In the hot plate assay, flies jump quickly to escape from a hot copper plate (>45 degrees C). Our results demonstrate that, as in mammals, the latency of the jump response is inversely related to stimulus intensity, and innoxious thermosensation does not elicit this nociceptive behavior. To explore the genetic mechanisms of nociception, we examined several mutants in both assays. Abnormal nociceptive behavior of a mutant, painless, indicates that painless, a gene essential for nociception in Drosophila larvae, is also required for thermal nociception in adult flies. painless is expressed in certain neurons of the peripheral nervous system and thoracic ganglia, as well as in the definite brain structures, the mushroom bodies. However, chemical or genetic insults to the mushroom bodies do not influence the nociceptive behavior, suggesting that different painless-expressing neurons play diverse roles in thermal nociception. Additionally, no-bridge(KS49), a mutant that has a structural defect in the protocerebral bridge, shows defective response to noxious heat. Thus, our results validate adult Drosophila as a useful model to study the genetic mechanisms of thermal nociception. PMID- 17081266 TI - Independent quantitative trait loci influence ventral and dorsal hippocampal volume in recombinant inbred strains of mice. AB - Anatomical and functional studies support segregation of the hippocampus into ventral and dorsal components along its septotemporal axis. However, it is unknown whether the development of these two components of the hippocampus is influenced by common or separate genetic factors. In this study, we used recombinant inbred strains of mice to determine whether the same or different quantitative trait loci (QTL) influence ventral and dorsal hippocampal volume. Using two sets of strains of recombinant inbred mice (BXD and AXB/BXA), we identified separate QTLs for ventral and dorsal hippocampal volume. In BXD mice, suggestive QTLs for ventral hippocampus were identified on chromosomes 2, 8 and 13, and a significant QTL for dorsal hippocampal volume was identified on chromosome 15. There was also a suggestive QTL for dorsal hippocampal volume on chromosome 13. In AXB/BXA mice, there were no significant or suggestive QTLs for ventral hippocampal volume, but a significant QTL for dorsal hippocampus was identified on chromosome 5. These findings suggest that the development of the ventral and dorsal components of the hippocampus is influenced by separate genetic loci. PMID- 17081267 TI - Cutaneous malignant lymphomas: update 2006. AB - Cutaneous lymphomas represent a unique group of lymphomas and are the second most frequent extranodal lymphomas. As with other neoplasias, the pathogenesis is based mainly on a stepwise accumulation of mutations of suppressor genes and oncogenes caused by genetic, environmental or infectious factors. The diagnostic work-up includes clinical, histological, imaging and hematological investigations and in many cases immunohistochemical and molecular biological analyses. The current WHO/EORTC classification of cutaneous lymphomas differentiates "mature T cell and NK-cell lymphomas", "mature B-cell lymphomas" and "immature hematopoietic malignancies", their variants and subgroups. It is compatible with the WHO classification for neoplasias of the hematopoietic and lymphoid tissue and respects the organ-specific peculiarities of primary cutaneous lymphomas. The assignment of the various types of cutaneous lymphomas into prognostic categories (pre-lymphomatous "abortive" disorders; definite malignant lymphomas of low-grade malignancy; definite malignant lymphomas of high-grade malignancy) provides essential information on the biological behavior and allows an appropriate planning of the therapeutic strategy, which may be topical or systemic and aggressive or non-aggressive. Besides the classical options for therapy, there are new and "experimental" strategies, the efficacy of which has to be studied in clinical trials. PMID- 17081268 TI - Xerosis and callus formation as a key to the diabetic foot syndrome: dermatologic view of the problem and its management. AB - The diabetic foot syndrome is a major complication of diabetes mellitus. The two most important pathophysiologic factors are peripheral arterial occlusion and peripheral neuropathy. The cutaneous lesion is a plantar ulcer, often accompanied by soft tissue and bone infections which can require amputation. Triggers include poorly fitting shoes, poor foot care, or overlooked foreign bodies, often coupled with a structural foot deformity. Increased plantar pressure, especially beneath the metatarsal heads, and the resultant callus play an important role. The patients often already have xerosis of the plantar skin with scales, fissures, erosions and impaired barrier function, complicating the situation. Prompt neurologic and vascular diagnostic studies, coupled with routine examination of the feet and primary prophylactic measures are most important. The most important therapeutic goals are optimal control of the diabetes mellitus, relieving pressure points and avoiding or reducing callus formation. PMID- 17081269 TI - Efficacy and safety of pimecrolimus cream 1% in adult patients with vitiligo: results of a randomized, double-blind, vehicle-controlled study. AB - BACKGROUND: Vitiligo is an acquired, pigmentary skin disorder which is disfiguring and difficult to treat. In an earlier open label study in adult patients with vitiligo, pimecrolimus cream 1% was reported to have similar efficacy as clobetasol propionate 0.05%. We performed a double-blind, intrapatient comparison of pimecrolimus cream 1% with placebo cream. PATIENTS AND METHODS: Twenty adult Caucasians with symmetrical vitiligo (predominantly on extremities, none in the face) were treated b.i.d. for 6 months left/right with pimecrolimus/vehicle (N = 10) or vehicle/pimecrolimus (N = 10), respectively. Primary efficacy endpoint was the size of the target lesion at month 6 and secondary efficacy endpoint was re-pigmentation. RESULTS: Treatment with pimecrolimus cream 1% or vehicle resulted in no significant change in mean target lesion size. Modest repigmentation (1-25%) was noted with pimecrolimus at month 2 in 12 of 17 patients (vehicle: 9 of 17 patients). Afterwards, the number of patients who experienced an improvement of pigmentation steadily decreased (3 of 14 patients with pimecrolimus and 2 of 14 with placebo at month 6). Treatment was well tolerated. There were no treatment-related adverse events, no induction of skin atrophy nor any other application site side effects. CONCLUSION: In this group of adult patients with symmetrical vitiligo, treatment of body lesions (except face) with pimecrolimus cream 1% could not be shown to be effective. PMID- 17081270 TI - Clinical Experience Acquired with Raptiva (CLEAR) trial in patients with moderate to-severe plaque psoriasis: results from extended treatment in an international, Phase III, placebo-controlled trial. AB - BACKGROUND: The 12-week, double-blind, placebo-controlled, first-treatment (FT) CLEAR trial period demonstrated the efficacy/safety of efalizumab in moderate-to severe plaque psoriasis, including refractory or contraindicated patients unsuitable for other systemic treatments. This study assessed the efficacy/safety of open-label extended treatment (up to 24 weeks' continuous treatment) in patients not achieving > or =75% improvement in Psoriasis Area and Severity Index (PASI-75) at week 12 of the FT period. Time to relapse after treatment cessation, and efficacy/safety of 12 weeks' open-label re-treatment in patients achieving PASI-75 at week 12 FT were also assessed. PATIENTS AND METHODS: Patients with PASI-75 at week 12 FT were observed without treatment until relapse, then re treated with open-label efalizumab (1.0 mg/kg/week for 12 weeks). Others received open-label extended treatment without intervening observation. RESULTS: Among efalizumab-treated patients (n = 308) who had < 75% PASI improvement at week 12 FT, extended treatment led to PASI-75 in 26.6%. Among patients with between > or = 50 and < 75% PASI improvement at week 12 FT (n = 118), 47.5% improved to PASI 75 with extended treatment. For patients achieving PASI-75 at week 12 FT (n = 164), median time to relapse was 58 days. Re-treatment after relapse led to mean PASI improvement of 62.3% from study baseline (n = 145). Safety results were consistent with previous studies, with no new safety concerns. CONCLUSIONS: These results demonstrate additional benefit of continuing efalizumab. Re-treatment re established disease control in patients with PASI-75 who relapsed following treatment cessation. The safety profile remained consistent with that seen at 12 weeks. PMID- 17081271 TI - Lupus vulgaris--a case diagnosed more than 20 years after onset. AB - A 47-year-old Turkish woman developed an erythematous scaly plaque on her thigh over 20 years. Cutaneous sarcoidosis had been diagnosed eight years previously on a skin biopsy. We confirmed Mycobacterium tuberculosis infection by polymerase chain reaction and cultivation of the causative organism from lesional skin. After several months of antituberculous treatment with isoniazid, rifampicin, ethambutol and pyrazinamide, the lesion resolved. PMID- 17081272 TI - Successful thalidomide therapy for actinic prurigo in a European woman. AB - Actinic prurigo is a rare, often difficult-to-treat, idiopathic photodermatosis. Actinic prurigo is divided into a hereditary form appearing in the Native American population and a sporadic form occurring in non-Native Americans. We present a 28-year-old Caucasian woman who developed typical clinical signs and symptoms of actinic prurigo, just as had her mother and grandmother. The patient and her mother were HLA-A24 and HLA-DR 4 with the subtype HLA-DRB1*0408. Based on clinical symptoms and the HLA pattern, the diagnosis of actinic prurigo was made. Treatment with thalidomide led to resolution of the disease. This case report of a Caucasian woman suffering from a hereditary form of actinic prurigo questions the established classification of actinic prurigo into a hereditary Native American form and a sporadic form occurring in the non-Native American population. PMID- 17081273 TI - Diagnostic approach to photodermatoses. PMID- 17081274 TI - [Erythematous lesion of the perianal area in a 75-year old woman]. PMID- 17081275 TI - [Removal of cysts using a punch technique]. PMID- 17081276 TI - Perianal squamous cell carcinoma. PMID- 17081277 TI - Abstracts of the 1st World Congress of Teledermatology & Annual Meeting of the Austrian Scientific Society of Telemedicine, Graz, Austria, November 9 - 11, 2006. PMID- 17081278 TI - Transformation. AB - The Nobel process for science are often somewhat controversial for who they omit. A posthumous Nobel honor could help recognize some neglected heroes. PMID- 17081279 TI - The tree of one percent. AB - Two significant evolutionary processes are fundamentally not tree-like in nature- lateral gene transfer among prokaryotes and endosymbiotic gene transfer (from organelles) among eukaryotes. To incorporate such processes into the bigger picture of early evolution, biologists need to depart from the preconceived notion that all genomes are related by a single bifurcating tree. PMID- 17081280 TI - Heparan sulfate proteoglycan-dependent neutrophil chemotaxis toward PR-39 cathelicidin. AB - Cathelicidins are mammalian proteins containing a C-terminal cationic antimicrobial domain. Porcine PR-39 cathelicidin affects leukocyte biology. Mechanisms of action may involve alteration of heparan sulfate proteoglycan dependent functions in inflammatory cells. It was tested whether PR-39 affects human neutrophil migration and if such effects involve heparan sulphate proteoglycans. Neutrophils were from forearm venous blood of healthy donors. Migration was tested in modified Boyden chamber assays. Involvement of heparan sulfate proteoglycans was tested by their chemical modification and by the use of specific antibodies. PR-39 induced migration in neutrophils in a concentration dependent manner. Modification of heparan sulfate proteoglycans with sodium chlorate inhibited migration whereas chemotaxis toward the chemoattractant formyl Met-Leu-Phe was not affected. Removal of heparan sulfates or chondroitin sulfates from the surface of neutrophils by heparinase or chondroitinase inhibited migration toward PR-39. In conclusion, antimicrobial PR-39 stimulates human neutrophil chemotaxis in a heparan sulfate proteoglycan-dependent manner. Involvement of syndecans is likely as both heparinase and chondroitinase were abrogating. Data suggest active participation of heparan sulfate proteoglycans of neutrophils in cathelicidin peptide-mediated regulation of the antimicrobial host defense. PMID- 17081281 TI - Qualitative evaluation of a form for standardized information exchange between orthopedic surgeons and occupational physicians. AB - BACKGROUND: Both occupational physicians and orthopedic surgeons can be involved in the management of work relevant musculoskeletal disorders. These physicians hardly communicate with each other and this might lead to different advice to the patient. Therefore, we evaluated a standardized information exchange form for the exchange of relevant information between the orthopedic surgeon and the occupational physician. The main goals of this qualitative study are to evaluate whether the form improved information exchange, whether the form gave relevant information, and to generate ideas to further improve this information exchange. METHODS: The information exchange form was developed in two consensus meetings with five orthopedic surgeons and five occupational physicians. To evaluate the information exchange form, a qualitative evaluation was set up. Structured telephone interviews were undertaken with the patients, interviews with the physicians were face-to-face and semi-structured, based on a topic list. These interviews were recorded and literally transcribed. Each interview was analyzed separately in Atlas-Ti. RESULTS: The form was used for 8 patients, 7 patients agreed to participate in the qualitative evaluation. All three orthopedic surgeons involved and three of the six involved occupational physicians agreed to be interviewed. The form was transferred to 4 occupational physicians, the other 3 patients recovered before they visited the occupational physician. The information on the form was regarded to be useful. All orthopedic surgeons agreed that the occupational physician should take the initiative. Most physicians felt that the form should not be filled out for each patient visiting an orthopedic surgeon, but only for those patients who do not recover as expected. Orthopedic surgeons suggested that a copy of the medical information provided to the general practitioner could also be provided to occupational physicians. CONCLUSION: The information exchange form was regarded to be useful and could be used in practice. The occupational physician should take the initiative for using this form and most physicians felt the information should only be exchanged for patients who do not recover as expected. That means that the advantage of giving information early in the treatment is lost. PMID- 17081282 TI - Reduction of nosocomial pneumonia after major burns by trace element supplementation: aggregation of two randomised trials. AB - INTRODUCTION: Nosocomial pneumonia is a major source of morbidity and mortality after severe burns. Burned patients suffer trace element deficiencies and depressed antioxidant and immune defences. This study aimed at determining the effect of trace element supplementation on nosocomial or intensive care unit (ICU)-acquired pneumonia. METHODS: Two consecutive, randomised, double-blinded, supplementation studies including two homogeneous groups of 41 severely burned patients (20 placebo and 21 intervention) admitted to the burn centre of a university hospital were combined. Intervention consisted of intravenous trace element supplements (copper 2.5 to 3.1 mg/day, selenium 315 to 380 mug/day, and zinc 26.2 to 31.4 mg/day) for 8 to 21 days versus placebo. Endpoints were infections during the first 30 days (predefined criteria for pneumonia, bacteraemia, wound, urine, and other), wound healing, and length of ICU stay. Plasma and skin (study 2) concentrations of selenium and zinc were determined on days 3, 10, and 20. RESULTS: The patients, 42 +/- 15 years old, were burned on 46% +/- 19% of body surface: the combined characteristics of the patients did not differ between the groups. Plasma trace element concentrations and antioxidative capacity were significantly enhanced with normalisation of plasma selenium, zinc, and glutathione peroxidase concentrations in plasma and skin in the trace element supplemented group. A significant reduction in number of infections was observed in the supplemented patients, which decreased from 3.5 +/- 1.2 to 2.0 +/- 1.0 episodes per patient in placebo group (p < 0.001). This was related to a reduction of nosocomial pneumonia, which occurred in 16 (80%) patients versus seven (33%) patients, respectively (p < 0.001), and of ventilator-associated pneumonia from 13 to six episodes, respectively (p = 0.023). CONCLUSION: Enhancing trace element status and antioxidant defences by selenium, zinc, and copper supplementation was associated with a decrease of nosocomial pneumonia in critically ill, severely burned patients. PMID- 17081283 TI - Antiinflammatory evaluation of leaves of Plumeria acuminata. AB - BACKGROUND: [corrected] Plumeria acuminata belonging to the family Apocynaceae is commonly known as 'perungalli' in Tamil and is widely distributed throughout the Southern parts of India. In traditional medicinal system different parts of the plant have been mentioned to be useful in a variety of diseases. The plant material is widely used as a purgative, remedy for diarrhoea and cure for itch. The milky juice is employed for the treatment of inflammation and rheumatism. The bark has been reported to be useful in hard tumors, diarrhoea and gonorrhoea. The objective of the present study was to evaluate the antiinflammatory activity of methanol extract of leaves of Plumeria acuminata on carrageenan, dextran, histamine and serotonin-induced inflammation in rat hind paw oedema models. METHODS: Acute and chronic inflammation models were used to evaluate the anti inflammatory activity of the extract. Wistar albino rats of either sex weighing 180-200 g were used. In acute model carrageenan, dextran, histamine and serotonin models were used to induce inflammation in rat hind paw and cotton pellet-induced granuloma method was used for chronic inflammation model. In each model four groups of six animals were used. In all the models Group I served as control (0.9% normal saline, 5 mlkg(-1) b.w) and group IV as standard (Indomethacin 10 mgkg(-1) b.w). Group II and III received extract at the doses of 250 and 500 mgkg(-1) b.w respectively. RESULTS: The methanol extract of Plumeria acuminata exhibited significant anti-inflammatory activity on the tested experimental animal models. The extract (500 mgkg(-1) b.w) exhibited maximum antiinflammatory effect i.e., 30.51, 47.06, 34.48 and 32.50% (P < 0.001) at the end of 3 h with carrageenan, dextran, histamine and serotonin respectively. Administration of MEPA (500 mgkg(-1) b.w) and indomethacin (10 mgkg(-1) b.w) significantly reduced the formation of granuloma tissue induced by cotton pellet method at a rate of 45.06 and 51.57% respectively. The effect produced by the extract was comparable to that of indomethacin a prototype of a nonsteroidal antiinflammatory agent. CONCLUSION: The results obtained in this study indicated that the methanol extract of Plumeria acuminata possess potent antiinflammatory activity in both acute and chronic models. PMID- 17081284 TI - Application of Petri net based analysis techniques to signal transduction pathways. AB - BACKGROUND: Signal transduction pathways are usually modelled using classical quantitative methods, which are based on ordinary differential equations (ODEs). However, some difficulties are inherent in this approach. On the one hand, the kinetic parameters involved are often unknown and have to be estimated. With increasing size and complexity of signal transduction pathways, the estimation of missing kinetic data is not possible. On the other hand, ODEs based models do not support any explicit insights into possible (signal-) flows within the network. Moreover, a huge amount of qualitative data is available due to high-throughput techniques. In order to get information on the systems behaviour, qualitative analysis techniques have been developed. Applications of the known qualitative analysis methods concern mainly metabolic networks. Petri net theory provides a variety of established analysis techniques, which are also applicable to signal transduction models. In this context special properties have to be considered and new dedicated techniques have to be designed. METHODS: We apply Petri net theory to model and analyse signal transduction pathways first qualitatively before continuing with quantitative analyses. This paper demonstrates how to build systematically a discrete model, which reflects provably the qualitative biological behaviour without any knowledge of kinetic parameters. The mating pheromone response pathway in Saccharomyces cerevisiae serves as case study. RESULTS: We propose an approach for model validation of signal transduction pathways based on the network structure only. For this purpose, we introduce the new notion of feasible t-invariants, which represent minimal self-contained subnets being active under a given input situation. Each of these subnets stands for a signal flow in the system. We define maximal common transition sets (MCT sets), which can be used for t-invariant examination and net decomposition into smallest biologically meaningful functional units. CONCLUSION: The paper demonstrates how Petri net analysis techniques can promote a deeper understanding of signal transduction pathways. The new concepts of feasible t-invariants and MCT-sets have been proven to be useful for model validation and the interpretation of the biological system behaviour. Whereas MCT-sets provide a decomposition of the net into disjunctive subnets, feasible t-invariants describe subnets, which generally overlap. This work contributes to qualitative modelling and to the analysis of large biological networks by their fully automatic decomposition into biologically meaningful modules. PMID- 17081285 TI - Does a joint development and dissemination of multidisciplinary guidelines improve prescribing behaviour: a pre/post study with concurrent control group and a randomised trial. AB - BACKGROUND: It is difficult to keep control over prescribing behaviour in general practices. The purpose of this study was to assess the effects of a dissemination strategy of multidisciplinary guidelines on the volume of drug prescribing. METHODS: The study included two designs, a quasi-experimental pre/post study with concurrent control group and a random sample of GPs within the intervention group. The intervention area with 53 GPs was compared with a control group of 54 randomly selected GPs in the south and centre of the Netherlands. Additionally, a randomisation was executed in the intervention group to create two arms with 27 GPs who were more intensively involved in the development of the guideline and 26 GPs in the control group. A multidisciplinary committee developed prescription guidelines. Subsequently these guidelines were disseminated to all GPs in the intervention region. Additional effects were studied in the subgroup trial in which GPs were invited to be more intensively involved in the guideline development procedure. The guidelines contained 14 recommendations on antibiotics, asthma/COPD drugs and cholesterol drugs. The main outcome measures were prescription data of a three-year period (one year before and 2 years after guideline dissemination) and proportion of change according to recommendations. RESULTS: Significant short-term improvements were seen for one recommendation: mupirocin. Long-term changes were found for cholesterol drug prescriptions. No additional changes were seen for the randomised controlled study in the subgroup. GPs did not take up the invitation for involvement. CONCLUSION: Disseminating multidisciplinary guidelines that were developed within a region, has no clear effect on prescribing behaviour even though GPs and specialists were involved more intensively in their development. Apparently, more effort is needed to bring about change. PMID- 17081286 TI - Knowledge and attitudes about health research amongst a group of Pakistani medical students. AB - BACKGROUND: Health research training is an important part of medical education. This study was conducted to assess the level of knowledge and attitudes regarding health research in a group of Pakistani medical students at Aga Khan University, Karachi. METHODS: It was a cross-sectional pilot study conducted among a group of Pakistani medical students. Through stratified random sampling, a pre-tested, structured and validated questionnaire was administered to 220 medical students. Knowledge and attitudes were recorded on a scale (graduated in percentages). RESULTS: Mean scores of students were 49.0% on knowledge scale and 53.7% on attitude scale. Both knowledge and attitudes improved significantly with increasing years of study in medical college [Regression coefficient 4.10 (p value; 0.019) and 6.67 (p-value; < 0.001) for knowledge and attitudes, respectively]. CONCLUSION: Medical students demonstrate moderate level of knowledge and attitude towards health research. Intensive training in this regard is associated with significant improvement in knowledge and attitudes of students towards health research. PMID- 17081287 TI - In situ methods to localize transgenes and transcripts in interphase nuclei: a tool for transgenic plant research. AB - Genetic engineering of commercially important crops has become routine in many laboratories. However, the inability to predict where a transgene will integrate and to efficiently select plants with stable levels of transgenic expression remains a limitation of this technology. Fluorescence in situ hybridization (FISH) is a powerful technique that can be used to visualize transgene integration sites and provide a better understanding of transgene behavior. Studies using FISH to characterize transgene integration have focused primarily on metaphase chromosomes, because the number and position of integration sites on the chromosomes are more easily determined at this stage. However gene (and transgene) expression occurs mainly during interphase. In order to accurately predict the activity of a transgene, it is critical to understand its location and dynamics in the three-dimensional interphase nucleus. We and others have developed in situ methods to visualize transgenes (including single copy genes) and their transcripts during interphase from different tissues and plant species. These techniques reduce the time necessary for characterization of transgene integration by eliminating the need for time-consuming segregation analysis, and extend characterization to the interphase nucleus, thus increasing the likelihood of accurate prediction of transgene activity. Furthermore, this approach is useful for studying nuclear organization and the dynamics of genes and chromatin. PMID- 17081288 TI - SMC3 knockdown triggers genomic instability and p53-dependent apoptosis in human and zebrafish cells. AB - BACKGROUND: The structural maintenance of chromosome 3 (SMC3) protein is a constituent of a number of nuclear multimeric protein complexes that are involved in DNA recombination and repair in addition to chromosomal segregation. Overexpression of SMC3 activates a tumorigenic cascade through which mammalian cells acquire a transformed phenotype. This has led us to examine in depth how SMC3 level affects cell growth and genomic stability. In this paper the effect of SMC3 knockdown has been investigated. RESULTS: Mammalian cells that are SMC3 deficient fail to expand in a clonal population. In order to shed light on the underlying mechanism, experiments were conducted in zebrafish embryos in which cell competence to undergo apoptosis is acquired at specific stages of development and affects tissue morphogenesis. Zebrafish Smc3 is 95% identical to the human protein, is maternally contributed, and is expressed ubiquitously at all developmental stages. Antisense-mediated loss of Smc3 function leads to increased apoptosis in Smc3 expressing cells of the developing tail and notocord causing morphological malformations. The apoptosis and the ensuing phenotype can be suppressed by injection of a p53-specific MO that blocks the generation of endogenous p53 protein. Results in human cells constitutively lacking p53 or BAX, confirmed that a p53-dependent pathway mediates apoptosis in SMC3-deficient cells. A population of aneuploid cells accumulated in zebrafish embryos following Smc3-knockdown whereas in human cells the transient downregulation of SMC3 level lead to the generation of cells with amplified centrosome number. CONCLUSION: Smc3 is required for normal embryonic development. Its deficiency affects the morphogenesis of tissues with high mitotic index by triggering an apoptotic cascade involving p53 and the downstream p53 target gene bax. Cells with low SMC3 level display centrosome abnormalities that can lead to or are the consequence of dysfunctional mitosis and/or aneuploidy. Collectively the data support the view that SMC3 deficiency affects chromosomal stability leading to the activation of p53-dependent mitotic checkpoint. PMID- 17081289 TI - Novel metaheuristic for parameter estimation in nonlinear dynamic biological systems. AB - BACKGROUND: We consider the problem of parameter estimation (model calibration) in nonlinear dynamic models of biological systems. Due to the frequent ill conditioning and multi-modality of many of these problems, traditional local methods usually fail (unless initialized with very good guesses of the parameter vector). In order to surmount these difficulties, global optimization (GO) methods have been suggested as robust alternatives. Currently, deterministic GO methods can not solve problems of realistic size within this class in reasonable computation times. In contrast, certain types of stochastic GO methods have shown promising results, although the computational cost remains large. Rodriguez Fernandez and coworkers have presented hybrid stochastic-deterministic GO methods which could reduce computation time by one order of magnitude while guaranteeing robustness. Our goal here was to further reduce the computational effort without loosing robustness. RESULTS: We have developed a new procedure based on the scatter search methodology for nonlinear optimization of dynamic models of arbitrary (or even unknown) structure (i.e. black-box models). In this contribution, we describe and apply this novel metaheuristic, inspired by recent developments in the field of operations research, to a set of complex identification problems and we make a critical comparison with respect to the previous (above mentioned) successful methods. CONCLUSION: Robust and efficient methods for parameter estimation are of key importance in systems biology and related areas. The new metaheuristic presented in this paper aims to ensure the proper solution of these problems by adopting a global optimization approach, while keeping the computational effort under reasonable values. This new metaheuristic was applied to a set of three challenging parameter estimation problems of nonlinear dynamic biological systems, outperforming very significantly all the methods previously used for these benchmark problems. PMID- 17081290 TI - Death and suicide among former child and adolescent psychiatric patients. AB - BACKGROUND: Increased mortality rates among previous child and adolescent psychiatry (CAP) patients have been found in Scandinavian studies up to the 1980s. The suicide risk in this group has been estimated to be almost five times higher than expected. This article addresses two questions: Do Swedish CAP patients continue to risk premature death and what kind of information related to psychiatric symptoms and/or behavior problems can predict later suicide? METHODS: Hospital files, Sweden's census databases (including immigration and emigration) and administrative databases (including the Swedish Hospital Discharge register and the Persons Convicted of Offences register), and the Cause of Death register were examined to determine the mortality rate in a group of 1,400 former CAP inpatients and outpatients over a period of 12-33 years. Observed and expected numbers of deceased were calculated with the prospective method and the standardized mortality ratio (SMR) method. The relative risk or the risk ratio (RR) is presented with 95% confidence intervals (CIs). Significance level tests were made using two-by-two tables and chi-square tests. The Cox proportional hazards regression model was used for survival analysis. RESULTS: Twenty-four males and 14 females died. Compared with the general population, the standardized mortality ratio in this group of CAP patients was significantly higher in both sexes. Behavioral problems, school problems, and co-morbid alcohol or drug abuse and criminality (including alcohol-related crimes) were found to be important predictors. Thirty-two deaths were attributed to suicide, intoxication, drug overdose, or accident; one patient died of an alcohol abuse-related disorder, and five patients died of natural causes. Suicide was the most common cause of death, but only 2 of these 19 cases were initially admitted for attempted suicide. CONCLUSION: We suggest that suicide and death prevention among CAP patients may not be a psychiatric issue per se but a future function of society's juvenile social-welfare investments and juvenile-delinquency prevention programs. PMID- 17081291 TI - Socioeconomic differentials in premature mortality in Rome: changes from 1990 to 2001. AB - BACKGROUND: While socioeconomic inequalities in mortality have widened in many countries, evidence of social differentials is scarce in Southern Europe. We studied temporal changes in premature mortality across socioeconomic groups in Rome between 1990 and 2001. METHODS: We analysed all 126,511 death certificates of residents of Rome aged 0-74 years registered between 1990-2001. A 4-level census block index based on the 1991 census was used as an indicator of socioeconomic position (SEP). Using routine mortality data, standardised mortality rates (per 100,000 inhabitants) were calculated by SEP and gender for four time periods. Rate ratios were used to compare mortality by gender and age. RESULTS: Overall premature mortality decreased in both genders and in all socioeconomic groups; the change was greater in the highest socio-economic group. In both men and women, inequalities in mortality strengthened during the 1990s and appeared to stabilise at the end of the 20th century. However, for 60-74 year old women the gap continued to widen. CONCLUSION: Socioeconomic inequalities in health in Rome are still present at the beginning of the 21st century. Strategies to monitor the impact of SEP on mortality over time in different populations should be implemented to direct health policies. PMID- 17081292 TI - A Paleolithic diet confers higher insulin sensitivity, lower C-reactive protein and lower blood pressure than a cereal-based diet in domestic pigs. AB - BACKGROUND: A Paleolithic diet has been suggested to be more in concordance with human evolutionary legacy than a cereal based diet. This might explain the lower incidence among hunter-gatherers of diseases of affluence such as type 2 diabetes, obesity and cardiovascular disease. The aim of this study was to experimentally study the long-term effect of a Paleolithic diet on risk factors for these diseases in domestic pigs. We examined glucose tolerance, post challenge insulin response, plasma C-reactive protein and blood pressure after 15 months on Paleolithic diet in comparison with a cereal based swine feed. METHODS: Upon weaning twenty-four piglets were randomly allocated either to cereal based swine feed (Cereal group) or cereal free Paleolithic diet consisting of vegetables, fruit, meat and a small amount of tubers (Paleolithic group). At 17 months of age an intravenous glucose tolerance test was performed and pancreas specimens were collected for immunohistochemistry. Group comparisons of continuous variables were made by use of the t-test. P < 0.05 was chosen for statistical significance. Simple and multivariate correlations were evaluated by use of linear regression analysis. RESULTS: At the end of the study the Paleolithic group weighed 22% less and had 43% lower subcutaneous fat thickness at mid sternum. No significant difference was seen in fasting glucose between groups. Dynamic insulin sensitivity was significantly higher (p = 0.004) and the insulin response was significantly lower in the Paleolithic group (p = 0.001). The geometric mean of C-reactive protein was 82% lower (p = 0.0007) and intra arterial diastolic blood pressure was 13% lower in the Paleolithic group (p = 0.007). In evaluations of multivariate correlations, diet emerged as the strongest explanatory variable for the variations in dynamic insulin sensitivity, insulin response, C-reactive protein and diastolic blood pressure when compared to other relevant variables such as weight and subcutaneous fat thickness at mid sternum. There was no obvious immunohistochemical difference in pancreatic islets between the groups, but leukocytes were clearly more frequent in sampled pancreas from the Cereal group. CONCLUSION: This study in domestic pigs suggests that a Paleolithic diet conferred higher insulin sensitivity, lower C-reactive protein and lower blood pressure when compared to a cereal based diet. PMID- 17081293 TI - The Fatty Liver Index: a simple and accurate predictor of hepatic steatosis in the general population. AB - BACKGROUND: Fatty liver (FL) is the most frequent liver disease in Western countries. We used data from the Dionysos Nutrition & Liver Study to develop a simple algorithm for the prediction of FL in the general population. METHODS: 216 subjects with and 280 without suspected liver disease were studied. FL was diagnosed by ultrasonography and alcohol intake was assessed using a 7-day diary. Bootstrapped stepwise logistic regression was used to identify potential predictors of FL among 13 variables of interest [gender, age, ethanol intake, alanine transaminase, aspartate transaminase, gamma-glutamyl-transferase (GGT), body mass index (BMI), waist circumference, sum of 4 skinfolds, glucose, insulin, triglycerides, and cholesterol]. Potential predictors were entered into stepwise logistic regression models with the aim of obtaining the most simple and accurate algorithm for the prediction of FL. RESULTS: An algorithm based on BMI, waist circumference, triglycerides and GGT had an accuracy of 0.84 (95%CI 0.81-0.87) in detecting FL. We used this algorithm to develop the "fatty liver index" (FLI), which varies between 0 and 100. A FLI < 30 (negative likelihood ratio = 0.2) rules out and a FLI > or = 60 (positive likelihood ratio = 4.3) rules in fatty liver. CONCLUSION: FLI is simple to obtain and may help physicians select subjects for liver ultrasonography and intensified lifestyle counseling, and researchers to select patients for epidemiologic studies. Validation of FLI in external populations is needed before it can be employed for these purposes. PMID- 17081294 TI - Differential cooperation of oncogenes with p53 and Bax to induce apoptosis in rhabdomyosarcoma. AB - BACKGROUND: Deregulated expression of oncogenes such as MYC and PAX3-FKHR often occurs in rhabdomyosarcomas. MYC can enhance cell proliferation and apoptosis under specific conditions, whereas PAX3-FKHR has only been described as anti apoptotic. RESULTS: In order to evaluate how MYC and PAX3-FKHR oncogenes influenced p53-mediated apoptosis, rhabdomyosarcoma cells were developed to independently express MYC and PAX3-FKHR cDNAs. Exogenous wild-type p53 expression in MYC transfected cells resulted in apoptosis, whereas there was only a slight effect in those transfected with PAX3-FKHR. Both oncoproteins induced BAX, but BAX induction alone without expression of wild-type p53 was insufficient to induce apoptosis. Data generated from genetically modified MEFs suggested that expression of all three proteins; MYC, BAX and p53, was required for maximal cell death to occur. CONCLUSION: We conclude that cooperation between p53 and oncoproteins to induce apoptosis is dependent upon the specific oncoprotein expressed and that oncogene-mediated induction of BAX is necessary but insufficient to enhance p53-mediated apoptosis. These data demonstrate a novel relationship between MYC and p53-dependent apoptosis, independent of the ability of MYC to induce p53 that may be important in transformed cells other than rhabdomyosarcoma. PMID- 17081295 TI - Porcine reproductive and respiratory syndrome virus (PRRSV) infection spreads by cell-to-cell transfer in cultured MARC-145 cells, is dependent on an intact cytoskeleton, and is suppressed by drug-targeting of cell permissiveness to virus infection. AB - BACKGROUND: Porcine reproductive and respiratory syndrome virus (PRRSV) is the etiologic agent of PRRS, causing widespread chronic infections which are largely uncontrolled by currently available vaccines or other antiviral measures. Cultured monkey kidney (MARC-145) cells provide an important tool for the study of PRRSV replication. For the present study, flow cytometric and fluorescence antibody (FA) analyses of PRRSV infection of cultured MARC-145 cells were carried out in experiments designed to clarify viral dynamics and the mechanism of viral spread. The roles of viral permissiveness and the cytoskeleton in PRRSV infection and transmission were examined in conjunction with antiviral and cytotoxic drugs. RESULTS: Flow cytometric and FA analyses of PRRSV antigen expression revealed distinct primary and secondary phases of MARC-145 cell infection. PRRSV antigen was randomly expressed in a few percent of cells during the primary phase of infection (up to about 20-22 h p.i.), but the logarithmic infection phase (days 2 3 p.i.), was characterized by secondary spread to clusters of infected cells. The formation of secondary clusters of PRRSV-infected cells preceded the development of CPE in MARC-145 cells, and both primary and secondary PRRSV infection were inhibited by colchicine and cytochalasin D, demonstrating a critical role of the cytoskeleton in viral permissiveness as well as cell-to-cell transmission from a subpopulation of cells permissive for free virus to secondary targets. Cellular expression of actin also appeared to correlate with PRRSV resistance, suggesting a second role of the actin cytoskeleton as a potential barrier to cell-to-cell transmission. PRRSV infection and cell-to-cell transmission were efficiently suppressed by interferon-gamma (IFN-gamma), as well as the more-potent experimental antiviral agent AK-2. CONCLUSION: The results demonstrate two distinct mechanisms of PRRSV infection: primary infection of a relatively small subpopulation of innately PRRSV-permissive cells, and secondary cell-to-cell transmission to contiguous cells which appear non-permissive to free virus. The results also indicate that an intact cytoskeleton is critical for PRRSV infection, and that viral permissiveness is a highly efficient drug target to control PRRSV infection. The data from this experimental system have important implications for the mechanisms of PRRSV persistence and pathology, as well as for a better understanding of arterivirus regulation. PMID- 17081296 TI - Is FKBP5 a genetic marker of affective psychosis? A case control study and analysis of disease related traits. AB - BACKGROUND: A dysregulation of the hypothalamic-pituitary-adrenal (HPA) axis has been proposed as an important pathogenic factor in depression. Genetic variants of FKBP5, a protein of the HPA system modulating the glucocorticoid receptor, have been reported to be genetically associated with improved response to medical treatment and an increase of depressive episodes. METHODS: We examined three single nucleotide polymorphisms (SNPs) in FKBP5, rs4713916 in the proposed promoter region, rs1360780 in the second intron and rs3800373 in the 3' untranslated region (3'-UTR), in a case-control study of Caucasian origin (affective psychosis: n = 248; controls: n = 188) for genetic association and association with disease related traits. RESULTS: Allele and genotype frequencies of rs4713916, rs1360780 and rs3800373 were not significantly different between cases and controls. Two three-locus haplotypes, G-C-T and A-T-G, accounted for 86.2% in controls. Odds ratios were not increased between cases and controls, except the rare haplotype G-C-G (OR 6.81), representing 2.1% of cases and 0.3% of controls. The frequency of rs4713916AG in patients deviated from expected Hardy Weinberg equilibrium, the genotype AA at rs4713916 in monopolar depression (P = 0.011), and the two-locus haplotype rs1360780T--rs3800373T in the total sample (overall P = 0.045) were nominally associated with longer continuance of disease. CONCLUSION: Our data do not support a significant genetic contribution of FKBP5 polymorphisms and haplotypes to affective psychosis, and the findings are inconclusive regarding their contribution to disease-related traits. PMID- 17081297 TI - Hierarchical amino acid utilization and its influence on fermentation dynamics: rifamycin B fermentation using Amycolatopsis mediterranei S699, a case study. AB - BACKGROUND: Industrial fermentation typically uses complex nitrogen substrates which consist of mixture of amino acids. The uptake of amino acids is known to be mediated by several amino acid transporters with certain preferences. However, models to predict this preferential uptake are not available. We present the stoichiometry for the utilization of amino acids as a sole carbon and nitrogen substrate or along with glucose as an additional carbon source. In the former case, the excess nitrogen provided by the amino acids is excreted by the organism in the form of ammonia. We have developed a cybernetic model to predict the sequence and kinetics of uptake of amino acids. The model is based on the assumption that the growth on a specific substrate is dependent on key enzyme(s) responsible for the uptake and assimilation of the substrates. These enzymes may be regulated by mechanisms of nitrogen catabolite repression. The model hypothesizes that the organism is an optimal strategist and invests resources for the uptake of a substrate that are proportional to the returns. RESULTS: Stoichiometric coefficients and kinetic parameters of the model were estimated experimentally for Amycolatopsis mediterranei S699, a rifamycin B overproducer. The model was then used to predict the uptake kinetics in a medium containing cas amino acids. In contrast to the other amino acids, the uptake of proline was not affected by the carbon or nitrogen catabolite repression in this strain. The model accurately predicted simultaneous uptake of amino acids at low cas concentrations and sequential uptake at high cas concentrations. The simulated profile of the key enzymes implies the presence of specific transporters for small groups of amino acids. CONCLUSION: The work demonstrates utility of the cybernetic model in predicting the sequence and kinetics of amino acid uptake in a case study involving Amycolatopsis mediterranei, an industrially important organism. This work also throws some light on amino acid transporters and their regulation in A. mediterranei. Further, cybernetic model based experimental strategy unravels formation and utilization of ammonia as well as its inhibitory role during amino acid uptake. Our results have implications for model based optimization and monitoring of other industrial fermentation processes involving complex nitrogen substrate. PMID- 17081298 TI - Attachment as an organizer of behavior: implications for substance abuse problems and willingness to seek treatment. AB - BACKGROUND: Attachment theory allows specific predictions about the role of attachment representations in organizing behavior. Insecure attachment is hypothesized to predict maladaptive emotional regulation whereas secure attachment is hypothesized to predict adaptive emotional regulation. In this paper, we test specific hypotheses about the role of attachment representations in substance abuse/dependence and treatment participation. Based on theory, we expect divergence between levels of maladaptive functioning and adaptive methods of regulating negative emotions. METHODS: Participants for this study consist of a sample of adoptees participating in an ongoing longitudinal adoption study (n = 208). The Semi-Structured Assessment of the Genetics of Alcohol-II 41 was used to determine lifetime substance abuse/dependence and treatment participation. Attachment representations were derived by the Adult Attachment Interview [AAI; 16]. We constructed a prior contrasts reflecting theoretical predictions for the association between attachment representations, substance abuse/dependence and treatment participation. RESULTS: Logistic regression was used to test our hypotheses. As predicted, individuals classified as dismissing, preoccupied or earned-secure reported the highest rates of substance abuse/dependence. Individuals classified as dismissing reported significantly lower rates of treatment participation despite their high rates of substance abuse/dependence. As expected, the continuous-secure group reported lowest rates of both substance abuse/dependence and treatment participation. CONCLUSION: The findings from this study identify attachment representations as an influential factor in understanding the divergence between problematic substance use and treatment utilization. The findings further imply that treatment may need to take attachment representations into account to promote successful recovery. PMID- 17081299 TI - What potential has tobacco control for reducing health inequalities? The New Zealand situation. AB - In this Commentary, we aim to synthesize recent epidemiological data on tobacco and health inequalities for New Zealand and present it in new ways. We also aim to describe both existing and potential tobacco control responses for addressing these inequalities. In New Zealand smoking prevalence is higher amongst Maori and Pacific peoples (compared to those of "New Zealand European" ethnicity) and amongst those with low socioeconomic position (SEP). Consequently the smoking related mortality burden is higher among these populations. Regarding the gap in mortality between low and high socioeconomic groups, 21% and 11% of this gap for men and women was estimated to be due to smoking in 1996-99. Regarding the gap in mortality between Maori and non-Maori/non-Pacific, 5% and 8% of this gap for men and women was estimated to be due to smoking. The estimates from both these studies are probably moderate underestimates due to misclassification bias of smoking status. Despite the modest relative contribution of smoking to these gaps, the absolute number of smoking-attributable deaths is sizable and amenable to policy and health sector responses. There is some evidence, from New Zealand and elsewhere, for interventions that reduce smoking by low-income populations and indigenous peoples. These include tobacco taxation, thematically appropriate mass media campaigns, and appropriate smoking cessation support services. But there are as yet untried interventions with major potential. A key one is for a tighter regulatory framework that could rapidly shift the nicotine market towards pharmaceutical-grade nicotine (or smokeless tobacco products) and away from smoked tobacco. PMID- 17081300 TI - Genomic presence of recombinant porcine endogenous retrovirus in transmitting miniature swine. AB - The replication of porcine endogenous retrovirus (PERV) in human cell lines suggests a potential infectious risk in xenotransplantation. PERV isolated from human cells following cocultivation with porcine peripheral blood mononuclear cells is a recombinant of PERV-A and PERV-C. We describe two different recombinant PERV-AC sequences in the cellular DNA of some transmitting miniature swine. This is the first evidence of PERV-AC recombinant virus in porcine genomic DNA that may have resulted from autoinfection following exogenous viral recombination. Infectious risk in xenotransplantation will be defined by the activity of PERV loci in vivo. PMID- 17081301 TI - 3D geometric reconstruction of thoracic aortic aneurysms. AB - BACKGROUND: The thoracic aortic aneurysm (TAA) is a pathology that involves an expansion of the aortic diameter in the thoracic aorta, leading to risk of rupture. Recent studies have suggested that internal wall stress, which is affected by TAA geometry and the presence or absence of thrombus, is a more reliable predictor of rupture than the maximum diameter, the current clinical criterion. Accurate reconstruction of TAA geometry is a crucial step in patient specific stress calculations. METHODS: In this work, a novel methodology was developed, which combines data from several sets of magnetic resonance (MR) images with different levels of detail and different resolutions. Two sets of images were employed to create the final model, which has the highest level of detail for each component of the aneurysm (lumen, thrombus, and wall). A reference model was built by using a single set of images for comparison. This approach was applied to two patient-specific TAAs in the descending thoracic aorta. RESULTS: The results of finite element simulations showed differences in stress pattern between the coarse and fine models: higher stress values were found with the coarse model and the differences in predicted maximum wall stress were 30% for patient A and 11% for patient B. CONCLUSION: This paper presents a new approach to the reconstruction of an aneurysm model based on the use of several sets of MR images. This enables more accurate representation of not only the lumen but also the wall surface of a TAA taking account of intraluminal thrombus. PMID- 17081302 TI - The financial losses from the migration of nurses from Malawi. AB - BACKGROUND: The migration of health professionals trained in Africa to developed nations has compromised health systems in the African region. The financial losses from the investment in training due to the migration from the developing nations are hardly known. METHODS: The cost of training a health professional was estimated by including fees for primary, secondary and tertiary education. Accepted derivation of formula as used in economic analysis was used to estimate the lost investment. RESULTS: The total cost of training an enrolled nurse midwife from primary school through nurse-midwifery training in Malawi was estimated as US$ 9,329.53. For a degree nurse-midwife, the total cost was US$ 31,726.26. For each enrolled nurse-midwife that migrates out of Malawi, the country loses between US$ 71,081.76 and US$ 7.5 million at bank interest rates of 7% and 25% per annum for 30 years respectively. For a degree nurse-midwife, the lost investment ranges from US$ 241,508 to US$ 25.6 million at 7% and 25% interest rate per annum for 30 years respectively. CONCLUSION: Developing countries are losing significant amounts of money through lost investment of health care professionals who emigrate. There is need to quantify the amount of remittances that developing nations get in return from those who migrate. PMID- 17081303 TI - Assessment of health care needs and utilization in a mixed public-private system: the case of the Athens area. AB - BACKGROUND: Given the public-private mix of the Greek health system, the purpose of this study was to assess whether variations in the utilisation of health services, both primary and inpatient care, were associated with underlying health care needs and/or various socio-economic factors. METHODS: Data was obtained from a representative sample (N = 1426) residing in the broader Athens area (response rate 70.6%). Perceived health-related quality of life (HRQOL), as measured by the physical and mental summary component scores of the SF-36 Health Survey, was used as a proxy of health care need. Health care utilization was measured by a) last month visits to public sector physicians, b) last-month visits to private sector physicians, c) last-year visits to hospital emergency departments and d) last year hospital admissions. Statistical analysis involved the implementation of logistic regression models. RESULTS: Health care need was the factor most strongly associated with all measures of health care utilization, except for visits to public physicians. Women, elderly, less wealthy and individuals of lower physical health status visited physicians contracted to their insurance fund (public sector). Women, well educated and those once again of lower physical health status were more likely to visit private providers. Visits to hospital emergency departments and hospital admissions were related to need and no socio economic factor was related to the use of those types of care. CONCLUSION: This study has demonstrated a positive relationship between health care need and utilisation of health services within a mixed public-private health care system. Concurrently, interesting differences are evident in the utilization of various types of services. The results have potential implications in health policy making and particularly in the proper allocation of scarce health resources. PMID- 17081304 TI - Myb proteins inhibit fibroblast transformation by v-Rel. AB - Genes that cause cancer have been divided into two general classes--oncogenes that act in a dominant fashion to transform normal cells into a malignant state, and tumor suppressor genes that act in a dominant fashion to prevent such transformation. In this report, we demonstrate that both the v-myb retroviral oncogene, which causes leukemic transformation of hematopoietic cells, and the c myb proto-oncogene can also function as inhibitors of fibroblast transformation by the v-rel oncogene. These results imply that the myb genes can function either as oncogenes or as tumor suppressors in different cellular contexts. PMID- 17081305 TI - Pathway-specific differences between tumor cell lines and normal and tumor tissue cells. AB - BACKGROUND: Cell lines are used in experimental investigation of cancer but their capacity to represent tumor cells has yet to be quantified. The aim of the study was to identify significant alterations in pathway usage in cell lines in comparison with normal and tumor tissue. METHODS: This study utilized a pathway specific enrichment analysis of publicly accessible microarray data and quantified the gene expression differences between cell lines, tumor, and normal tissue cells for six different tissue types. KEGG pathways that are significantly different between cell lines and tumors, cell lines and normal tissues and tumor and normal tissue were identified through enrichment tests on gene lists obtained using Significance Analysis of Microarrays (SAM). RESULTS: Cellular pathways that were significantly upregulated in cell lines compared to tumor cells and normal cells of the same tissue type included ATP synthesis, cell communication, cell cycle, oxidative phosphorylation, purine, pyrimidine and pyruvate metabolism, and proteasome. Results on metabolic pathways suggested an increase in the velocity nucleotide metabolism and RNA production. Pathways that were downregulated in cell lines compared to tumor and normal tissue included cell communication, cell adhesion molecules (CAMs), and ECM-receptor interaction. Only a fraction of the significantly altered genes in tumor-to-normal comparison had similar expressions in cancer cell lines and tumor cells. These genes were tissue-specific and were distributed sparsely among multiple pathways. CONCLUSION: Significantly altered genes in tumors compared to normal tissue were largely tissue specific. Among these genes downregulation was a major trend. In contrast, cell lines contained large sets of significantly upregulated genes that were common to multiple tissue types. Pathway upregulation in cell lines was most pronounced over metabolic pathways including cell nucleotide metabolism and oxidative phosphorylation. Signaling pathways involved in adhesion and communication of cultured cancer cells were downregulated. The three way pathways comparison presented in this study brings light into the differences in the use of cellular pathways by tumor cells and cancer cell lines. PMID- 17081306 TI - Effects of self-management intervention on health outcomes of patients with heart failure: a systematic review of randomized controlled trials. AB - BACKGROUND: Heart failure is the most common cause of hospitalization among adults over 65. Over 60% of patients die within 10 years of first onset of symptoms. The objective of this study is to determine the effectiveness of self management interventions on hospital readmission rates, mortality, and health related quality of life in patients diagnosed with heart failure. METHODS: The study is a systematic review of randomized controlled trials. The following data sources were used: MEDLINE (1966-11/2005), EMBASE (1980-11/2005), CINAHL (1982 11/2005), the ACP Journal Club database (to 11/2005), the Cochrane Central Trial Registry and the Cochrane Database of Systematic Reviews (to 11/2005); article reference lists; and experts in the field. We included randomized controlled trials of self-management interventions that enrolled patients 18 years of age or older who were diagnosed with heart failure. The primary outcomes of interest were all-cause hospital readmissions, hospital readmissions due to heart failure, and mortality. Secondary outcomes were compliance with treatment and quality of life scores. Three reviewers independently assessed the quality of each study and abstracted the results. For each included study, we computed the pooled odds ratios (OR) for all-cause hospital readmission, hospital readmission due to heart failure, and death. We used a fixed effects model to quantitatively synthesize results. We were not able to pool effects on health-related quality of life and measures of compliance with treatment, but we summarized the findings from the relevant studies. We also summarized the reported cost savings. RESULTS: From 671 citations that were identified, 6 randomized trials with 857 patients were included in the review. Self-management decreased all-cause hospital readmissions (OR 0.59; 95% confidence interval (CI) 0.44 to 0.80, P = 0.001) and heart failure readmissions (OR 0.44; 95% CI 0.27 to 0.71, P = 0.001). The effect on mortality was not significant (OR = 0.93; 95% CI 0.57 to 1.51, P = 0.76). Adherence to prescribed medical advice improved, but there was no significant difference in functional capabilities, symptom status and quality of life. The reported savings ranged from 1300 to 7515 dollars per patient per year. CONCLUSION: Self management programs targeted for patients with heart failure decrease overall hospital readmissions and readmissions for heart failure. PMID- 17081307 TI - Evolution and comparative analysis of the MHC Class III inflammatory region. AB - BACKGROUND: The Major Histocompatibility Complex (MHC) is essential for immune function. Historically, it has been subdivided into three regions (Class I, II, and III), but a cluster of functionally related genes within the Class III region has also been referred to as the Class IV region or "inflammatory region". This group of genes is involved in the inflammatory response, and includes members of the tumour necrosis family. Here we report the sequencing, annotation and comparative analysis of a tammar wallaby BAC containing the inflammatory region. We also discuss the extent of sequence conservation across the entire region and identify elements conserved in evolution. RESULTS: Fourteen Class III genes from the tammar wallaby inflammatory region were characterised and compared to their orthologues in other vertebrates. The organisation and sequence of genes in the inflammatory region of both the wallaby and South American opossum are highly conserved compared to known genes from eutherian ("placental") mammals. Some minor differences separate the two marsupial species. Eight genes within the inflammatory region have remained tightly clustered for at least 360 million years, predating the divergence of the amphibian lineage. Analysis of sequence conservation identified 354 elements that are conserved. These range in size from 7 to 431 bases and cover 15.6% of the inflammatory region, representing approximately a 4-fold increase compared to the average for vertebrate genomes. About 5.5% of this conserved sequence is marsupial-specific, including three cases of marsupial-specific repeats. Highly Conserved Elements were also characterised. CONCLUSION: Using comparative analysis, we show that a cluster of MHC genes involved in inflammation, including TNF, LTA (or its putative teleost homolog TNF-N), APOM, and BAT3 have remained together for over 450 million years, predating the divergence of mammals from fish. The observed enrichment in conserved sequences within the inflammatory region suggests conservation at the transcriptional regulatory level, in addition to the functional level. PMID- 17081308 TI - Incorporating genome-scale tools for studying energy homeostasis. AB - Mammals have evolved complex regulatory systems that enable them to maintain energy homeostasis despite constant environmental challenges that limit the availability of energy inputs and their composition. Biological control relies upon intricate systems composed of multiple organs and specialized cell types that regulate energy up-take, storage, and expenditure. Because these systems simultaneously perform diverse functions and are highly integrated, they are extremely difficult to understand in terms of their individual component contributions to energy homeostasis. In order to provide improved treatments and clinical options, it is important to identify the principle genetic and molecular components, as well as the systemic features of regulation. To begin, many of these features can be discovered by integrating experimental technologies with advanced methods of analysis. This review focuses on the analysis of transcriptional data derived from microarrays and how it can complement other experimental techniques to study energy homeostasis. PMID- 17081309 TI - Predictors of betel quid chewing behavior and cessation patterns in Taiwan aborigines. AB - BACKGROUND: Betel quid, chewed by about 600 million people worldwide, is one of the most widely used addictive substances. Cessation factors in betel quid chewers are unknown. The present study explores prevalence and the quit rate of betel quid chewing in Taiwan aborigines. Our goal was to delineate potential predictors of chewing cessation. METHODS: A stratified random community-based survey was designed for the entire aborigines communities in Taiwan. A total of 7144 participants were included between June 2003 and May 2004 in this study. Information on sociodemographic characteristics, such as gender, age, obesity, education years, marital status, ethnicity, and habits of betel quid chewing, smoking and drinking was collected by trained interviewers. RESULTS: The prevalence of betel quid chewers was 46.1%. Betel quid chewing was closely associated with obesity (OR = 1.61; 95% CI: 1.40-1.85). Betel quid chewers were most likely to use alcohol and cigarettes together. Quit rate of betel quid chewers was 7.6%. Betel quid chewers who did not drink alcohol were more likely to quit (OR = 1.89; 95% CI: 1.43-2.50). Alcohol use is a significant factor related to cessation of betel quid chewing, but smoking is not. CONCLUSION: Taiwan aborigines have a high prevalence of betel quid chewers and a low quit rate. Alcohol use is strongly association with betel quid chewing. Efforts to reduce habitual alcohol consumption might be of benefit in cessation of betel quid chewing. PMID- 17081310 TI - Multidisciplinary approach to diagnosis and management of osteosarcoma - a review of the St Vincent's Hospital experience. AB - BACKGROUND: Osteosarcoma is the most common primary malignant bone tumour in children and young adults. Despite advances in the diagnosis and management of osteosarcoma, there have been few recent studies describing the experiences of tertiary referral centres. This paper aims to describe and discuss the clinical features, pre-operative work-up, management and outcomes of these patients at St Vincent's Hospital (Melbourne, Australia). METHODS: Retrospective study of fifty nine consecutive patients managed for osteosarcoma at St Vincent's Hospital between 1995 and 2005. RESULTS: Median age at diagnosis was 21 (range, 11-84) years. Gender distribution was similar, with thirty-one male and twenty-eight female patients.Twenty-five patients had osteosarcoma in the femur, eleven each were located in the humerus and tibia, six were identified in the pelvis, and one each in the clavicle, maxilla, fibula, sacrum, ulna and radius.Pre-operative tissue diagnosis of osteosarcoma was obtained through computed tomography-guided percutaneous biopsy in over ninety percent of patients. Following initial therapy, over fifty percent of patients remained relapse-free during the follow up period, with twelve percent and twenty-seven percent of patients documented as having local and distant disease recurrence, respectively. Of patients with recurrent disease, sixty-two percent remained disease-free following subsequent surgical intervention (most commonly, pulmonary metastatectomy). CONCLUSION: Patient outcomes can be optimised through a multidisciplinary approach in a tertiary referral centre. At St Vincent's Hospital, survival and relapse rates of patients managed for osteosarcoma compare favourably with the published literature. PMID- 17081311 TI - Spatial and temporal variation in malaria transmission in a low endemicity area in northern Tanzania. AB - BACKGROUND: Spatial and longitudinal monitoring of transmission intensity will allow better targeting of malaria interventions. In this study, data on meteorological, demographic, entomological and parasitological data over the course of a year was collected to describe malaria epidemiology in a single village of low transmission intensity. METHODS: Entomological monitoring of malaria vectors was performed by weekly light trap catches in 10 houses. Each house in the village of Msitu wa Tembo, Lower Moshi, was mapped and censused. Malaria cases identified through passive case detection at the local health centre were mapped by residence using GIS software and the incidence of cases by season and distance to the main breeding site was calculated. RESULTS: The principle vector was Anopheles arabiensis and peak mosquito numbers followed peaks in recent rainfall. The entomological inoculation rate estimated was 3.4 (95% CI 0.7-9.9) infectious bites per person per year. The majority of malaria cases (85/130) occurred during the rainy season (chi2 = 62,3, p < 0.001). Living further away from the river (OR 0.96, CI 0.92-0.998, p = 0.04 every 50 m) and use of anti-insect window screens (OR 0.65, CI 0.44-0.94, p = 0.023) were independent protective factors for the risk of malaria infection. Children aged 1-5 years and 5-15 years were at greater risk of clinical episodes (OR 2.36, CI 1.41-3.97, p = 0.001 and OR 3.68, CI 2.42-5.61, p < 0.001 respectively). CONCLUSION: These data show that local malaria transmission is restricted to the rainy season and strongly associated with proximity to the river. Transmission reducing interventions should, therefore, be timed before the rain-associated increase in mosquito numbers and target households located near the river. PMID- 17081312 TI - The effects of incomplete protein interaction data on structural and evolutionary inferences. AB - BACKGROUND: Present protein interaction network data sets include only interactions among subsets of the proteins in an organism. Previously this has been ignored, but in principle any global network analysis that only looks at partial data may be biased. Here we demonstrate the need to consider network sampling properties explicitly and from the outset in any analysis. RESULTS: Here we study how properties of the yeast protein interaction network are affected by random and non-random sampling schemes using a range of different network statistics. Effects are shown to be independent of the inherent noise in protein interaction data. The effects of the incomplete nature of network data become very noticeable, especially for so-called network motifs. We also consider the effect of incomplete network data on functional and evolutionary inferences. CONCLUSION: Crucially, when only small, partial network data sets are considered, bias is virtually inevitable. Given the scope of effects considered here, previous analyses may have to be carefully reassessed: ignoring the fact that present network data are incomplete will severely affect our ability to understand biological systems. PMID- 17081313 TI - A statistical score for assessing the quality of multiple sequence alignments. AB - BACKGROUND: Multiple sequence alignment is the foundation of many important applications in bioinformatics that aim at detecting functionally important regions, predicting protein structures, building phylogenetic trees etc. Although the automatic construction of a multiple sequence alignment for a set of remotely related sequences cause a very challenging and error-prone task, many downstream analyses still rely heavily on the accuracy of the alignments. RESULTS: To address the need for an objective evaluation framework, we introduce a statistical score that assesses the quality of a given multiple sequence alignment. The quality assessment is based on counting the number of significantly conserved positions in the alignment using importance sampling method in conjunction with statistical profile analysis framework. We first evaluate a novel objective function used in the alignment quality score for measuring the positional conservation. The results for the Src homology 2 (SH2) domain, Ras-like proteins, peptidase M13, subtilase and beta-lactamase families demonstrate that the score can distinguish sequence patterns with different degrees of conservation. Secondly, we evaluate the quality of the alignments produced by several widely used multiple sequence alignment programs using a novel alignment quality score and a commonly used sum of pairs method. According to these results, the Mafft strategy L-INS-i outperforms the other methods, although the difference between the Probcons, TCoffee and Muscle is mostly insignificant. The novel alignment quality score provides similar results than the sum of pairs method. CONCLUSION: The results indicate that the proposed statistical score is useful in assessing the quality of multiple sequence alignments. PMID- 17081314 TI - Animals and their products utilized as medicines by the inhabitants surrounding the Ranthambhore National Park, India. AB - The present ethnozoological study describes the traditional knowledge related to the use of different animals and animal-derived products as medicines by the inhabitants of villages surrounding the Ranthambhore National Park of India (Bawaria, Mogya, Meena), which is well known for its very rich biodiversity. The field survey was conducted from May to July 2005 by performing interviews through structured questionnaires with 24 informants (16 men and 8 women), who provided information regarding therapeutic uses of animals. A total of 15 animals and animal products were recorded and they are used for different ethnomedical purposes, including tuberculosis, asthma, paralysis, jaundice, earache, constipation, weakness, snake poisoning. The zootherapeutic knowledge was mostly based on domestic animals, but some protected species like the collared dove (Streptopelia sp.), hard shelled turtle (Kachuga tentoria), sambhar (Cervus unicolor) were also mentioned as important medicinal resources. We would suggest that this kind of neglected traditional knowledge should be included into the strategies of conservation and management of faunistic resources in the investigated area. PMID- 17081315 TI - Own and parental war experience as a risk factor for mental health problems among adolescents with an immigrant background: results from a cross sectional study in Oslo, Norway. AB - BACKGROUND: An increasing proportion of immigrants to Western countries in the past decade are from war affected countries. The aim of this study was to estimate the prevalence of war experience among adolescents and their parents and to investigate possible differences in internalizing and externalizing mental health problems between adolescents exposed and unexposed to own and parental war experience. METHOD: The study is based on a cross-sectional population-based survey of all 10th grade pupils in Oslo for two consecutive years. A total of 1,758 adolescents were included, all with both parents born outside of Norway. Internalizing and externalizing mental health problems were measured by Hopkins Symptom Checklist-10 and subscales of the Strengths and Difficulties Questionnaire, respectively. Own and parental war experience is based on adolescent self-report. RESULTS: The proportion of adolescents with own war experience was 14% with the highest prevalence in immigrants from Eastern Europe and Sub-Saharan Africa. The proportion of parental war experience was 33% with Sub-Saharan Africa being highest. Adolescents reporting own war experience had higher scores for both internalizing and externalizing mental health problems compared to immigrants without war experience, but only externalizing problems reached statistically significant differences. For parental war experience there was a statistically significant relationship between parental war experience and internalizing mental health problems. The association remained significant after adjustment for parental educational level and adolescents' own war experience. CONCLUSION: War exposure is highly prevalent among immigrants living in Oslo, Norway, both among adolescents themselves and their parents. Among immigrants to Norway, parental war experience appears to be stronger associated with mental health problems than adolescents own exposure to war experience. PMID- 17081316 TI - Lymphokine-activated killer cell susceptibility and adhesion molecule expression of multidrug resistant breast carcinoma. AB - Reports showing susceptibility of multidrug resistant (MDR) cancer cells to immune effectors, together with P-glycoprotein (P-gp) expression in immune effector subsets, including immature natural killer (NK) cells, and some activated T cells, suggest P-gp or some changes associated with it, have implications in immune-mediated mechanisms. A series of experiments were done to determine the nature of alterations associated with susceptibility to immune effector cells of MDR tumor cells. A cell line isolated from the malignant pleural effusion of a breast cancer patient was transfected with human and murine MDR1 genes, and four variants with different levels of MDR were obtained. Lymphokine-activated killer (LAK) activity was measured by a 51Chromium release, and conjugate formation assays. MDR1 transfectant P-gp+ breast carcinoma lines had increased LAK susceptibility compared to their parent line. Some part of the increased LAK susceptibility of drug-resistant cell lines was at the binding/recognition level as shown by conjugate formation assays. This suggests that differences may exist between paired cell lines with respect to the expression of cell adhesion molecules (CAMs). Monoclonal antibodies (mAbs) to CAMs and flow cytometry were used to quantitate these antigens. The CAMs studied were those previously found to be upregulated by stimulating NK cells with (interleukin-2) IL-2; ICAM-1 (CD54), LFA-3 (CD58), N-CAM (CD56), and the beta chain of LFA-1 (CD18). Although no differences in these CAMs were found between the breast carcinoma line and its MDR1-transfected variants, the target susceptibility results given above suggest that IL-2 treatment could be effective in combination with current protocols using chemotherapeutics, monoclonal antibodies (mAbs) and stem cell transplantation. PMID- 17081318 TI - Laparoscopic surgery in urological oncology: brief overview. AB - The authors report the experience of a high-volume center with laparoscopic surgery in urological oncology, as well as a review of other relevant series. Laparoscopic outcomes in the treatment of adrenal, kidney, upper tract transitional cell carcinoma, bladder, prostate, and testicular malignancy are described in this review. Specific considerations as complications and port-site recurrence are also addressed. The authors concluded that the intermediate-term oncological data is encouraging and comparable to open surgery. PMID- 17081319 TI - Transdermal drug delivery treatment for overactive bladder. AB - Overactive bladder is commonly treated with oral anticholinergic drugs such as oxybutynin chloride. Although oral anticholinergic agents have been effective in controlling urinary urgency and incontinence, adverse events, particularly dry mouth, often cause patients to discontinue oral therapy and to endure incontinence. Oxybutynin can be delivered transcutaneously, maintaining the efficacy of oral oxybutynin while significantly minimizing side effects (e.g., dry mouth) that may complicate therapy. By avoiding hepatic and gastrointestinal metabolism of oxybutynin, less N-desethyloxybutynin (N-DEO) is produced and this compound is deemed to be responsible for anticholinergic side effects such as dry mouth. This novel oxybutynin formulation offers patients with OAB and urge urinary incontinence a well-tolerated option for managing the symptoms of overactive bladder. PMID- 17081320 TI - Retroperitoneoscopic nephrectomy in benign pathology. AB - INTRODUCTION: We report our experience with 43 retroperitoneal laparoscopic nephrectomy for benign kidney disease. MATERIALS AND METHODS: All patients had a poor function from obstructive uropathology and renal atrophy. None of these patients had a previous lumbotomy. Retroperitoneoscopy was performed with 4 trocar port technique in a lateral position. The retroperitoneal space is created by using a Gaur's balloon made of sterile glove. The approach to vascular pedicle was done posteriorly and vessels were clipped by metal and Hem-o-lock (Weck Closure Systems, North Carolina, USA) clips. The sample was intact extracted in an Endo-Bag prolonging one trocar incision. RESULTS: Median operative time was 160 minutes and median blood loss was 200 mL. Four cases (9%) were converted to open surgery: one case due to bleeding and 3 cases due to technical difficulties regarding perirenal adherences. Most patients (39) checked out from the Hospital in day two. Four of them were left over 3 days due to wound complications. CONCLUSIONS: Retroperitoneoscopy offers a safe, effective and reproductive access to nephrectomy for benign pathologies. PMID- 17081321 TI - Orthotopic ileal neobladder reconstruction for bladder cancer: is adjuvant chemotherapy safe? AB - OBJECTIVE: We examined our database of patients undergoing radical cystectomy (RC) with orthotopic neobladder (NB) to determine whether adjuvant chemotherapy in this group is safe. MATERIALS AND METHODS: We performed a retrospective analysis of patients who underwent radical cystectomy and urinary diversion between 1992 and 2004. Relevant clinical and therapeutic data were entered into a database. High-risk bladder cancer patients who underwent NB were identified. They were stratified into 2 groups, those who received adjuvant chemotherapy and those who did not. The incidence of complications between the 2 groups was analyzed and compared. RESULTS: Over the 12-year period, 136 patients underwent RC and NB construction for bladder cancer. Of these, 83 patients were at high risk for recurrence. Nineteen patients received adjuvant chemotherapy and 64 did not. The complication rate in the adjuvant chemotherapy group was 53% and it was 23% in those who did not receive chemotherapy. There were no perioperative or treatment related death. There were 2 patients with grade 4 toxicity in the adjuvant chemotherapy group. There was a statistical difference between these two groups with regard to the incidence of complications. However, none of these complications was life-threatening, required only conservative treatment and caused no long-term disability. CONCLUSIONS: Adjuvant chemotherapy is a safe treatment for patients undergoing RC and NB substitution. Hence, the option of orthotopic NB should not be denied in selected bladder cancer patients with high risk for recurrent disease. PMID- 17081322 TI - A fourteen-year review of radical cystectomy for transitional cell carcinoma demonstrating the usefulness of the concept of lymph node density. AB - OBJECTIVE: We studied the long-term outcome of radical cystectomy for transitional cell carcinoma and evaluated prognostic factors for disease specific survival. MATERIALS AND METHODS: A retrospective review was carried out for all cystectomies performed for transitional cell carcinoma between 1989 and 2002. Disease specific survival was correlated to patient, pathological and operative factors as well as to adjuvant therapy. RESULTS: Of the 133 cystectomies included, 100 were male and 33 were female patients. The median age was 69 years (range 43 to 86). The median follow up was 20 months (range 0 to 158). With univariate analysis, pT stage, N stage, lymph node density, carcinoma in-situ, surgical margin and post-operative radiotherapy to distant metastasis were predictive of disease specific survival. On the other hand, with multivariate analysis, only pT stage, lymph node density and post-operative radiotherapy to distant metastasis were predictive of disease specific survival. Within the group of node positive disease, lymph node density also predicted disease specific survival with both univariate and multivariate analyses. Patients with lymph node density 20% or below showed better disease specific survival. CONCLUSIONS: pT stage and lymph node density were found to be the most important predictive factors for disease specific survival after cystectomy in the Asian population. PMID- 17081323 TI - Salvage radical prostatectomy: an alternative treatment for local recurrence of radioresistant cancer. AB - OBJECTIVES: The treatment of recurrent prostate cancer after radiotherapy or brachytherapy through radical prostatectomy has been little indicated due to the concern over the procedure's morbidity. We present the experience of our service with postradiotherapy radical prostatectomy. MATERIALS AND METHODS: Between 1996 and 2002, 9 patients submitted to radiotherapy due to prostate cancer were treated with salvage surgery for locally recurrent disease. All patients had a biopsy of the prostate confirming the tumor recurrence, increase in the PSA levels and staging without evidence of a systemic disease. We have assessed the morbidity and the recurrence-free survival rate after salvage radical prostatectomy. RESULTS: Preradiotherapy PSA varied from 6.2 to 50 ng/mL (mean 17.3) and clinical staging T1, T2 and T3 in 33.3%, 44.4% and 22.2% of the patients respectively. The interval for the biopsy after conforming external beam radiotherapy or brachytherapy varied from 8 to 108 months (median: 36). Four patients received antiandrogenic therapy neoadjuvant to the surgery with a mean of 7 months (1-48) after radiotherapy. From the six patients potent before the surgery, three have presented erectile dysfunction. Urinary incontinence as well as bladder neck sclerosis occurred in two patients (22.2%). Biochemical recurrence occurred in two individuals (22.2%) 12 months after the surgery. Biochemical recurrence-free survival rate was 77.8% with median follow-up time of 30 months (8-102). CONCLUSION: Salvage radical prostatectomy is a safe and effective alternative for the treatment of locally recurrent prostate cancer after radiotherapy and brachytherapy. PMID- 17081324 TI - Eggshell calcification of kidney in ureteropelvic junction obstruction. AB - Eggshell calcification of kidney in case of ureteropelvic junction obstruction (UPJO) is an uncommon finding with only a few cases reported in literature. We report a thirty-year-old symptomatic man with curvilinear calcification in hydronephrotic right kidney. Thorough investigations to rule out genitourinary tuberculosis and hydatid disease of kidney were performed prior to the definitive management by laparoscopic approach. PMID- 17081325 TI - Complete en bloc urinary exenteration for synchronous multicentric transitional cell carcinoma with sarcomatoid features in a hemodialysis patient. AB - The incidence of transitional cell carcinoma (TCC) in patients submitted to hemodialysis is low. The presence of TCC with sarcomatoid features in this cohort is even scarcer. Herein, we describe a very rare case of synchronous multicentric muscle invasive bladder carcinoma with prostate invasion in a hemodialysis patient, submitted to complete en bloc urinary exenteration. PMID- 17081326 TI - Sarcomatoid carcinoma with osseous differentiation in the bladder. AB - INTRODUCTION: Bladder sarcomatoid carcinoma is a very rare variant of transitional cell carcinoma. With disputed nomenclature, the tumor has been described previously under a variety of names such as sarcomatoid carcinoma, pseudosarcoma, malignant mixed mesodermal/Mullerian tumor, metaplastic carcinoma and spindle cell carcinoma. This malignancy represents 0.3% of all bladder tumors and has an aggressive behavior yielding a poor prognosis despite radio and chemotherapy. CASE REPORT: An 81 y/o man presented with a transitional cell carcinoma and underwent a transurethral resection. Adjuvant onco-BCG was introduced. After 9 months of follow-up, a local tumoral recurrence occurred and a new transurethral resection revealed sarcomatoid carcinoma with osseous elements. A radical cystoprostatectomy was then carried out. PMID- 17081327 TI - Small round blue cell tumor of seminal vesicle in a young patient. AB - Seminal vesicle tumor is a rare disease with unclear origin. Generally, it is presented as a pelvic mass that can be detected by sonography and digital rectal exam. The authors report a 25-year-old patient with a pelvic mass which the magnetic resonance and surgical specimen reveal a seminal vesicle tumor. Immunohistochemical findings favored a primitive neuroectodermal tumor of the seminal vesicle. Herein, the treatment, histological and histochemical findings of this entity are discussed. PMID- 17081328 TI - Early successful orchidopexy does not prevent from developing azoospermia. AB - INTRODUCTION: The incidence of Ad spermatogonia (stem cells for fertility) was assessed in 20 cryptorchid patients, all of whom had a successful orchidopexy in childhood but developed azoospermia following puberty. MATERIALS AND METHODS: From a cohort of 231 patients who had a semen analysis following successful orchidopexy 20 patients (9%) had azoospermia. The patients were classified into 2 groups according to the time of surgery: A = < 21 months of age (n = 5, mean = 10.7 +/- 8.6 months) and B = during childhood (n = 15, mean = 10.1 +/- 3 years). Nine of the 20 patients (45%) had bilateral cryptorchidism: A = 1 and B = 8. Testicular biopsies were performed during orchidopexy and analyzed with semi-thin technique. The number of Ad spermatogonia and entire number of germ cells was determined. The patients' semen analyses were evaluated at least twice; FSH and testosterone plasma values were estimated. RESULTS: In group A, all patients had germ cells at the time of surgery (mean = 1.04 +/- 1.4 germ cells per tubular cross section); only 6 patients in group B (40%) had no germ cells (mean = 0.17 +/- 0.4); A vs. B, p = 0.0133. Importantly, Ad spermatogonia were absent in the entire study population. The plasma FSH of 16 patients (80%) was abnormal [median = 16.35 IU/L (Interquartile range of sample - IQR 9.075-27.85 95% CI, 3-53)] while the plasma testosterone of all the patients was normal. CONCLUSIONS: The most severe cause of infertility in cryptorchid patients cannot be mitigated by an early successful surgery alone. PMID- 17081329 TI - Ureteral calcinosis in juvenile dermatomyositis: successful precocious surgical management. AB - We report a successful surgical intervention to repair bilateral ureteral strictures in a child with juvenile dermatomyositis (JDM) and ureteral calcinosis. This is the fourth reported case in medical literature. A 9-year-old girl with severe JDM, a rare connective tissue disease characterized by skin and muscles vasculitis, was under immunosuppressive therapy. In the course of the disease, she presented recurrent urinary tract infections. Bilateral ureteral dilation was detected by ultrasound (US) and intravenous pyelogram (IVP). CT scan showed bilateral ureteral calculus. Ureteroscopy revealed bilateral ureteral calcinosis, confirmed by histopathological analysis. Bilateral double-J stents were placed, resulting in transient improvement of ureteral dilation and infection, but only the surgical removal of abnormal ureteral portions was successful. In conclusion, endourological approach is recommended for diagnosis of urinary tract involvement by JDM because radiological evaluation can be misleading. The immunosuppressive treatment and the resection of damaged ureteral segments have allowed the control of urinary complications. PMID- 17081330 TI - An exploration into patient preference for injectable therapy over surgery in the treatment of female urinary incontinence. AB - OBJECTIVE: To explore patient preference for injectable therapy over open surgery in the treatment of urinary incontinence. MATERIAL AND METHODS: Fifty-eight female patients presented for treatment of urinary incontinence. During the initial interview process, they were asked to quantify their preference for injectable therapy over surgery by specifying the lowest success rate they would accept and still try injectable therapy. The results were summarized and assessed in relation to patient age and history of previous urogynecologic surgery. RESULTS: The mean lowest acceptable success rate for all 58 surveyed patients was 34%, with 23 (40%) accepting a success rate of only 10%. Although not statistically significant, the data suggested that older patients may tend to accept lower success rates than younger patients (mean of 39% for patients aged less than 60 years compared to 22% for those aged 80 years or older). There was no difference in response based on history of previous urogynecologic surgery. CONCLUSION: Patients appear willing to accept a relatively low success rate for injectable therapy compared to open surgery. PMID- 17081331 TI - Development of a urinary lithiasis localizer mechanism to couple ultrasound and extracorporeal lithotripsy equipment in canine model. AB - INTRODUCTION: Due to the evolution of extracorporeal lithotripsy equipment (ESWL) and presently, the fact that most part of the equipment does not present ultrasound to localize urinary calculi, a system that allows adapting ultrasound equipment to ESWL equipment was developed, disposing only of fluoroscopy. Thus, this equipment was developed and was tested in urinary stones in canine models, to check its precision in relation to fluoroscopy. METHOD: Seven male dogs were utilized with the introduction, in the bladder through the ureteral route, of chalkstones, with initial localization by fluoroscopy, with a further ultrasound coincidence check localization of the vesical stones, being submitted to ESWL with a 3-hour, 21 days and 60 days follow-up after the procedure. RESULTS: Success of localization in all animals was verified presenting elimination of stones in the first micturitions, after ESWL. No complications were verified in those animals for 60 days. CONCLUSION: We verified that this equipment can lead to an update of the equipment that use only fluoroscopy, increasing in this way, their technical capacity in the treatment of urinary calculi, mainly in cases of non-radiopaque stones. PMID- 17081353 TI - Effects of eccentric exercise on joint stiffness and muscle connectin (titin) isoform in the rat hindlimb. AB - We investigated the effects of repeated eccentric exercise for rat medial gastrocnemius muscle on ankle joint stiffness and muscle connectin (titin) isoform composition (longer form, alpha-connectin; shorter form, beta-connectin). Male Wistar rats were trained on a custom-made, isokinetic dynamometer (eccentric exercise group, n = 6; sham-operated group, n = 6). The exercise session consisted of 20 eccentric contractions elicited by submaximal electric stimulations under anesthesia. The contracting muscle was forcibly lengthened by an isokinetic dorsi-flexion of the ankle joint (velocity, 30 degrees/s; range of motion, 45 degrees). Rats in the eccentric-exercise group were trained every two days for 20 days (10 sessions in total). The static passive resistive torque (PRT) of 45 degrees at the ankle joint was used as a measure of the joint stiffness, and was determined before and after the experimental period. After 10 sessions of eccentric exercise, the wet weight of medial gastrocnemius muscle significantly increased (P < 0.05), whereas the static PRT significantly decreased (P < 0.05) in the eccentric-exercise group, when compared to the sham operated group. Myosin-ATPase staining showed a decrease in the number of type IIb/IId fibers (P < 0.001) and an increase in the number of type IIa fibers (P < 0.05). However, no significant difference was seen in the connectin (titin) isoform composition between the eccentric-exercise group and the sham-operated group, suggesting that the reduction in PRT was not due to change in resting mechanical properties of muscle fibers. PMID- 17081354 TI - Organelles containing inositol trisphosphate receptor type 2 in adrenal medullary cells. AB - To identify which organelles contained inositol trisphosphate (InsP(3)) receptor type 2 (InsP(3)R2) in adrenal medullary (AM) cells, immunocytochemical and biochemical studies were performed on AM cells of several species. InsP(3)R2-like immunoreactive materials produced by two different anti-InsP(3)R2 antibodies (Abs) (Chemicon and Sigma) were distributed in rat AM cells in agreement with BODIPY-FL-InsP(3) binding sites. For two other Abs (KM1083 and Santa Cruz), some of the anti-InsP(3)R2 immunoreactive materials were stained with an anti-dopamine beta-hydroxylase Ab, but not by BODIPY-FL-InsP(3). BODIPY-FL-thapsigargin binding sites were consistent with a distribution of the endoplasmic reticulum (ER) identified by an anti-calnexin Ab, and a prior application of thapsigargin significantly eliminated BODIPY-FL-thapsigargin bindings, suggesting that BODIPY FL-thapsigargin bindings were mediated by thapsigargin, but not the fluorescence molecule. The anti-InsP(3)R2 Ab that produced stainings consistent with BODIPY-FL InsP(3) bindings recognized a protein with about 250 kDa. A fractional analysis of bovine adrenal medullae revealed that the 250 kDa InsP(3)R2 was detected in a crude membrane fraction, but not in a secretory granule fraction. The results suggest that the InsP(3)R2 was present in the ER, but not in secretory granules in AM cells. PMID- 17081355 TI - [Observation on the safety: clinical trail on intracoronary autologous bone marrow mononuclear cells transplantation for acute myocardial infarction]. AB - OBJECTIVE: To investigate the safety of autologous bone marrow mononuclear cell (BM-MNCs) transplantation by intracoronary infusion in patients with acute myocardial infarction (AMI). METHODS: One hundred and eighty-four patients with AMI treated with percutaneous coronary intervention (PCI) were randomized in a 1:1 way to either intracoronary transplantation of autologous BM-MNCs (n = 92) right after PCI or to sodium chloride concluding heparin (controlled, n = 92) via a micro infusion catheter. In the process of the intracoronary infusion of BM MNCs, the complications should be recorded, which were aberration reflect (including of pale, syncope, nausea, hypotension and shock), deterioration of angina or heart failure, arrhythmias (including of bradycardia, sinus arrest or atrial ventricular block or ventricular fibrillation), embolism etc. Body temperature, blood pressure and heart rates should be monitored during the first week after transplantation. Holter, coronary angiography and ultrasonic cardiography were performed at the designed time points. Main heart accidents, restenosis and tumor were recorded during 2-years follow up. RESULTS: During the period of bone marrow puncture and intracoronary infusion of BM-MNCs, few patients occurred pale, dizziness, bradycardia and hypotension, which were transient and due to vagus reflect. No stem cell-related arrhythmias, deterioration of angina were noted. In BM-MNCs group one patient developed in stent reocclusion in one week after transplantation, five developed in-stent restenosis during further follow-up 30 months, which were similar with control group. There were no deaths, major adverse cardiac events, tumor and other late adverse events during follow-up period in both groups. CONCLUSION: Intracoronary transplantation of autologous BM-MNCs in the acute phase after AMI is feasible and seems safe in the 30 months of follow-up. PMID- 17081356 TI - [The autologous bone marrow mononuclear cell transplantation by intracoronary route treat patients with severe heart failure after myocardial infarction]. AB - OBJECTIVE: To investigate the chronic effects of intracoronary autologous bone marrow mononuclear cell (BM-MNCs) transplantation in patients with refractory heart failure (RIHF) after myocardial infarction. METHODS: Thirty patients with RIHF (LVEF < 40%) were enrolled in this nonrandomized study, autologous BM-MNCs (5.0 +/- 0.7) x 10(7) were transplanted with via infarct-related coronary artery in 16 patients and 14 patients received standard medical therapy served as control. Baseline and follow up evaluations included complete clinical evaluations, plasma BNP, ANP, ET-1 measurements, echocardiography, PET, and Holter monitoring. RESULTS: Baseline characteristics were similar between the 2 groups. There were no major periprocedural complications. One patient developed ventricular premature contractions during cell infusion for several seconds and recovered spontaneously. Compared to pre-transplantation, plasma BNP and ET-1 significantly decreased and plasma ANP significantly increased at 7 days post transplantation; 6 minutes walking distance increased from (72.1 +/- 31.5) to (201.6 +/- 23.3) m (P < 0.01), LVEF increased 9.9% (P < 0.001) and FDG-PET revealed vital myocardium area increased (10.3 +/- 3.4)% (P < 0.01) at 3 months after BM-MNCs transplantation. At 6 months follow up, the NYHA class improved from (3.4 +/- 0.1 to 2.4 +/- 0.2, P < 0.001) and no patient died and 1 patient rehospitalized due to lower extremities edema. In control group, LVEF decreased 7.2% compared to baseline (P < 0.001) and was significantly lower than transplantation group at 3 months (P < 0.001). At 6 months follow up, the NYHA class increased from (3.5 +/- 0.1 to 3.9 +/- 0.1, P < 0.05), 2 patients died and 10 patients rehospitalized due to aggravated heart failure. CONCLUSION: Present study demonstrates that intracoronary transplantation of autologous BM-MNCs is safe and effective for treating patients with RIHF after myocardial infarction. PMID- 17081357 TI - [Beneficial effects of human umbilical cord blood mononuclear cells transplantation on rats with experimental myocardial infarction]. AB - OBJECTIVE: Human umbilical cord blood contains abundant immature stem/progenitor cells, which may contribute to the repair of infarcted myocardium. Present study aimed to explore the feasibility and effects of human umbilical cord blood mononuclear cells (HUCBC) transplantation for the treatment of myocardial infarction. METHODS: Forty-five male Wistar rats were randomly divided into three groups: (1) Myocardial infarction (MI plus vehicle, n = 15), (2) MI plus cell transplantation (HUCBC were implanted into the peri-infarct area immediately after MI, n = 15), (3) Normal control group (n = 15). After echocardiography and hemodynamic measurements, the rats were sacrificed for histological and immunochemical examinations one month post MI. RESULTS: The transplanted HUCBC survived and participated the repair process in host heart. Significantly improved left ventricular function was evidenced by echocardiography in cell transplantation group compared to the MI control group. Left ventricular end diastolic pressure was significantly reduced LVEDP (21.08 +/- 8.10) vs (30.82 +/- 9.59) mm Hg, P < 0.05], +dp/dt(max) [(4.29 +/- 1.27) vs (3.24 +/- 0.75) mm Hg/ms, P < 0.05] and -dp/dt(max) increased [(3.71 +/- 0.79) vs (3.00 +/- 0.49) mm Hg/ms, P < 0.05] in cell transplantation rats compared with MI control rats. vWF immunostaining examination showed significantly increased microvessels within the boundary of infarcted myocardium in cell transplantation group compared to the MI control group (P < 0.01). CONCLUSIONS: HUCBC transplantation may improve cardiac function in MI rats by promoting microvessel formation. PMID- 17081359 TI - [The Chinese registry on reperfusion strategies and outcomes in ST-elevation myocardial infarction]. AB - OBJECTIVE: To analyze the current use of reperfusion strategies and the outcomes of patients with ST elevation acute coronary syndromes (ACS) in China. METHODS: A total of 518 consecutive patients (371 male and 147 females, mean age 65 +/- 11) with ST elevation ACS or newly discovered left bundle branch block were registered from 20 hospitals from 5 regions (ranging from large regional centre hospitals to small county hospitals) in China. Patient general characteristics, reperfusion patterns and outcomes were analyzed. Patients were followed up for 3 months. RESULTS: The median time from pain onset to presentation at the hospital was 4 hours. Pre-hospital delay > 12 hours was found in 20% patients. Fifty-six percent patients (292/518) underwent reperfusion therapy (134 with primary percutaneous coronary intervention and 158 with fibrinolysis). The median time from admission to reperfusion (door-to-needle) was 65 min in fibrinolysis group and 110 min (door-to-cath) in primary PCI group respectively. Urokinase was used in 67% (106/158) patients underwent fibrinolysis. Multivariate logistic regression analysis showed that age >/= 75 years (P < 0.01), previous myocardial infarction (P < 0.01) and history of congestive heart failure (P < 0.05) were associated with no reperfusion therapy. Mortality and congestive heart failure rates were significantly higher in patients with no reperfusion therapy not only at discharge (P < 0.01) but also at 3 months (P < 0.01) compared to patients underwent reperfusion. The incidence of combined outcomes (death or MI, and death, MI or Strobe) was also higher in patients without reperfusion therapy at 3 months (all P < 0.01) compared to patients underwent reperfusion. There were no differences on combined outcomes between fibrinolysis and primary PCI subgroups. CONCLUSION: Reperfusion therapy was the primary treatment of choice to improve the outcomes of patients with ST elevation ACS. Strategies to increase reperfusion therapy rate for ST elevation ACS are urgently needed in China. PMID- 17081360 TI - [Monocyte chemotactic protein-1 expression in coronary atherosclerosis plaque of sudden coronary death patients]. AB - OBJECTIVE: To investigate the expression of monocyte chemotactic protein 1 (MCP 1) in coronary atherosclerosis plaque of sudden coronary death (SCD) patients and the relationship between MCP-1 expression and SCD. METHODS: Autopsy heart samples (n = 90) collected during 2001 - 2003 were divided to SCD group (n = 36) and 2 control groups: control group I, non-SCD CHD (n = 28), control group II, non cardiac death (n = 26). The immuno-histochemistry SABC techniques (R, positive MCP-1 cell area/totl area) and computerized images analysis (A) were performed to detect the expression of MCP-1 in different groups. RESULTS: R and A in plaques are significant higher in SCD group than control group I and II (0.1264 +/- 0.013 vs 0.0269 +/- 0.0110 and 0.0267 +/- 0.0100, P = 0.04), (0.4534 +/- 0.083 vs 0.2303 +/- 0.040 and 0.2158 +/- 0.0400, P = 0.00), and similar between control group I and control group II. CONCLUSIONS: MCP-1 expression is increased in coronary atherosclerosis plaque of SCD patients. PMID- 17081361 TI - [Autoantibodies against alpha1 adrenergic receptor related with cardiac remodeling in hypertensive patients by clinical observation]. AB - OBJECTIVE: To investigate the effects of autoantibodies against alpha(-) adrenergic receptor on cardiac remodeling in patients with hypertension. METHODS: Five hundred and fifty three patients with hypertension in our hospital were selected. The autoantibodies against alpha(1) adrenergic receptor in sera of donor were detected by ELISA, and the results of echocardiography were recorded. By multiple logistic regressions, the risk factors were analyzed on left ventricular enlargement of hypertension. RESULTS: The percentage of autoantibodies against alpha(1) adrenergic receptor positive was 32.3% (179/553). There were significant difference between the positive group and negative group on the ratio of left atrial enlargement (53.6%, 44.3%, respectively; P < 0.05) and left ventricular enlargement (12.8%, 6.1%, respectively; P < 0.01). The result of regression analysis demonstrated that 4 risk factors were related to left ventricular enlargement, including male, course of disease, heart rate (HR) and autoantibodies against alpha(1) adrenergic receptor in the serum (all P < 0.05). CONCLUSIONS: The autoantibodies against alpha(1) adrenergic receptor have a relationship with left ventricular enlargement of hypertension. Patients with the activity of autoantibodies against alpha(1) adrenergic might contribute to predict cardiac remodeling. PMID- 17081362 TI - [The clinical efficiency and safety of bisoprolol hydrochlorothiazide in patients with mild to moderate essential hypertension]. AB - OBJECTIVE: To investigate the efficacy and safety of bisoprolol/hydrochlorothiazide (Lodoz) in patients with mild and moderate essential hypertension. METHODS: After 2 weeks of placebo run-in period, 90 hypertensive patients with sitting diastolic blood pressure (DBP) between 95 and 109 mm Hg (1 mm Hg = 0.133 kPa) and systolic blood pressure (SBP) below 180 mm Hg were treated by Lodoz (2.5 mg/6.25 mg/day) for 4 weeks. If DBP > 90 mm Hg at 4 weeks, Lodoz (5 mg/6.25 mg/day) was given for another 8 weeks. Clinic systolic and diastolic blood pressure measurements and ambulatory blood pressure monitoring (ABPM) were performed at the end of placebo run-in period and at 4 and 8 weeks. RESULTS: After 4 or 8 weeks treatment with Lodoz, clinic systolic and diastolic blood pressure, the 24-hour mean, daytime and nocturnal blood pressures reduced significantly compared to placebo run-in period [SBP and DBP reduced (14.89 +/- 10.99)/(10.37 +/- 7.35) mm Hg (4 weeks) and (19.40 +/- 10.55)/(13.31 +/- 7.77) mm Hg (8 weeks)] respectively (P < 0.05). The total efficacy rate is 59.3% for Lodoz 2.5 mg/6.25 mg and 69.8% for Lodoz 5 mg/6.25 mg. The trough: peak ratio for SBP and DBP were 91.5% and 94.4% with Lodoz 2.5 mg/6.25 mg, and 79.9% and 80.5% with Lodoz 5 mg/6.25 mg. The smoothness index (SI) for SBP and DBP were 9.07 and 6.48 with Lodoz 2.5 mg/6.25 mg, and 4.17 and 4.47 with Lodoz 5 mg/6.25 mg, respectively. Few side effects were observed during treatment including mild headache and dizziness and slightly increased serum urea acid. CONCLUSION: Lodoz (2.5 mg/6.25 mg and 5 mg/6.25 mg) can effectively reduce the 24 hours blood pressure in patients with mild to moderate essential hypertension. PMID- 17081363 TI - [Beneficial effects of probucol on endothelial function in patients with acute coronary syndrome]. AB - OBJECTIVE: The study investigate the antioxidant probucol on endothelial function in patients with acute coronary syndrome (ACS). METHODS: A total of 49 ACS patients randomly received standard therapy plus probucol (P, n = 24) or standard therapy (C, n = 25). Plasma oxidized low-density lipoprotein (ox-LDL), nitric oxide (NO) and circulating endothelial cells (CEC) were measured. The brachial arterial hyperemia-induced flow mediated dilation (FMD) and sublingual nitroglycerin (NTG) mediated vasodilatations were measured by high resolution ultrasound. These variables were analyzed before and after 3 months therapy. RESULTS: Plasma NO and FMD was significantly increased after 3 months therapy than before therapy [(80.46 +/- 10.24) micromol/Lvs (48.46 +/- 12.24) micromol/L, P < 0.01; (13.46 +/- 1.20)% vs (7.45 +/- 1.02)%, P < 0.05, respectively], while the number of CEC and ox-LDL were significantly decreased (P < 0.01) in P group. These values were similar before and after 3 months in C group. The linear correlation analysis showed that plasma ox-LDL negatively correlated with NO (r = -0.574, P < 0.01) and FMD (r = -0.517, P < 0.01) and positively correlated with CEC (r = 0.385, P < 0.01) in patients received 3 months probucol therapy. CONCLUSIONS: Chronic antioxidant probucol therapy could improve endothelial function in patients with ACS. PMID- 17081364 TI - [Mature-type adrenomedullin in coronary circulation immediately after reperfusion in patients with anterior acute myocardial infarction]. AB - OBJECTIVE: Levels of adrenomedullin (AM), a potent vasodilatory peptide, have been shown to increase in the early stage of acute myocardial infarction (AMI). The purpose of this study was to determine whether coronary sinus-aortic step-up of mature forms of AM is accelerated in patients with AMI after reperfusion. METHODS: The subjects were 146 consecutive patients with a first episode of anterior AMI and 51 normal controls. All patients with AMI underwent balloon reperfusion therapy within 24 h after symptom onset. Plasma levels of two molecular forms of AM (an active, mature form [AM-m] and an intermediate, inactive glycine-extended form [AM-Gly]) in the aorta and coronary sinus (CS) were measured by specific immunoradiometric assay after reperfusion. RESULTS: Plasma levels of AM-m and AM-Gly in the aorta and CS were higher in AMI patients than in controls. CS-aortic step-up of AM-m, which is an index of myocardial production of AM-m, was significantly greater in AMI patients than in controls [(1.7 +/- 1.4) pmol/L vs (0.4 +/- 0.3) pmol/L, P < 0.01]. However, there was no significant difference in CS-aortic step-up of AM-Gly (P = 0.30). AMI patients with left ventricular dysfunction (n = 49) had a significantly higher CS-aortic AM-m step-up than AMI patients without left ventricular dysfunction (n = 97). AMm in the aorta and CS negatively correlated with the left ventricular ejection fraction (r = -0.50, r = -0.48, P < 0.01). CONCLUSION: Myocardial synthesis of AM m is accelerated in patients with reperfused AMI, especially in patients with critical left ventricular dysfunction. Increased myocardial synthesis of active AM may protect against cardiac dysfunction, myocardial remodeling, or both after the onset of AMI. PMID- 17081365 TI - [Novel SCN5A gene mutations associated with Brugada syndrome: V95I, A1649V and delF1617]. AB - OBJECTIVE: Brugada syndrome is an inherited channelopathy that characterized by ST-segment elevation in the right precordial lead (V(1)-V(3)) on the electrocardiogram with or without right bundle branch block and related with high risk of sudden cardiac death and structurally normal hearts. The first and only gene linked to this disease is SCN5A, a gene encodes for alpha subunit of the cardiac sodium channel. The objective of this study is to explore SCN5A gene mutations in Chinese patients with Brugada syndrome. METHODS: Four patients diagnosed as Brugada syndrome and nine patients with suspected Brugada syndrome were chosen for the study. The exons in the functional regions of SCN5A gene were amplified with polymerase chain reaction and the amplified products were sequenced with Sanger method. If a mutation was identified, patient's family members were also screened. RESULTS: Two heterozygous mutations were found in one family diagnosed as Brugada syndrome. One missense mutation was a G-->A transition in the first nucleotide of codon 95 in SCN5A gene exon 3, which was predicted to result in substitution of Valine with Isoleucine (V95I). The other missense mutation was a C-->T transition in the second nucleotide of codon 1649 in SCN5A gene exon 28, which was predicted to result in substitution of Alanine with Valine (A1649V). A heterozygous mutation was identified in one family suspected to have the disease. The mutation was a three nucleotides (TCT) deletion that caused Phenylalanine deletion in codon 1617 in SCN5A gene exon 28. The three mutations were not detected in 100 control chromosomes. CONCLUSIONS: Mutation in SCN5A gene is one of the causes of Brugada syndrome in Chinese. Three novel SCN5A gene mutations were identified in Chinese with Brugada syndrome, which expands the spectrum of SCN5A mutations associated with the disease. PMID- 17081366 TI - [Endothelial progenitor cells related gene expression changes before and early after revascularization in patients with acute myocardial infarction]. AB - OBJECTIVE: The purpose of this study was to observe the endothelial progenitor cells (EPCs) related gene expression changes before and early after revascularization in patients with acute myocardial infarction. METHODS: Peripheral blood samples were taken from patients with acute anterior myocardial infarction 6 hours and 7 days after PCI and stenting. Mononuclear cells (MNCs) were isolated by Ficoll-density centrifugation and cultured in M-199 medium. After 14 days culture, attaching cells incorporated DiI-acetylated low-density lipoprotein (EPCs) were collected and RNA was isolated by Trizol for microarray analysis on 24 genes associated with permissibility/vessel tone (angiotensin system: ACE, AGTR-1, AGTR-2; NO system: eNOS; prostacyclin system: COX-2; endothelin system: ET-1, ETA, ETB; superoxide anions system: SOD-1), angiogenesis (adhesion molecule: CDH5; growth factors and receptors: VEGFR1, VEGFR2, VEGF) and endothelial cell activation (adhesion molecules expression: ICAM1, ICAM2, ICAM3, PECAM-1, E-Selectin, L-Selection, VCAM1; change phenotype from antithrombotic to prothrombotic: tPA, uPA, PAI, vWF). VEGFR2, PECAM-1 and VE-cadherin positive cells were identified by flow cytometry. RESULT: Eight gene expressions (AGTR-1, AGTR-2, COX-2, eNOS, ET-1, ETA, VEGF) were significantly downregulated 7 days post PCI compared to pre-PCI (P < 0.05). Flow cytometry results showed that VEGFR2 positive cells were also significantly reduced post PCI than that of before PCI (P < 0.05). CONCLUSION: PCI down-regulated endothelial progenitor cells related gene expressions in patients with acute myocardial infarction. PMID- 17081370 TI - [Combined use of transmyocardial stents with gene therapy in the treatment of acute myocardial infarction]. AB - OBJECTIVES: To determine the efficacy of combined use of transmyocardial stent with gene therapy to treat acute myocardial infarction in porcine model. METHODS: 24 Chinese mini swines have been divided into 4 groups randomly: group myocardial infarction (group MI n(1) = 6), group transmyocardial stent (group ST n(2) = 6), group vascular endothelial growth factor (group VEGF n(3) = 6), group transmyocardial stent and VEGF (group ST + VEGF n(4) = 6). In group MI, acute myocardial infarction animal model has been established by the ligation of the left descending coronary artery. In group ST, after the establishment of the model, 3 transmyocardial stents were implanted. In group VEGF, an expression plasmid containing the gene-encoding VEGF(165) (300 microg) was administered directly in the myocardium at 6 sites. In group ST + VEGF, both transmyocardial stents and expression plasmid containing the gene-encoding VEGF(165) are applied. 4 weeks later, the animals are sacrificed and echocardiography and pathological analysis have been done. RESULTS: The density of blood vessel in group ST, VEGF and ST + VEGF are significantly higher than group MI. And capillary density in group ST + VEGF is the highest in these groups statistically. Expression of VEGF was detected in group ST, VEGF and ST + VEGF, but in group VEGF and ST + VEGF the level of expression are higher. CONCLUSION: Combined use of transmyocardial stent with gene therapy has synthetic effect for the treatment of acute myocardial ischemia in porcine model and can significantly increase the vascular density. PMID- 17081371 TI - [The relationship between endothelin-1 and tumor necrosis factor-alpha and myocardial microcirculation dysfunction]. AB - OBJECTIVE: To investigate the relationship between endothelin-1 (ET-1) and tumor necrosis factor-alpha and myocardial microcircular dysfunction during coronary microembolization (CME). METHODS: CME was induced in 10 miniswine by selective infusion of microspheres (45 microm) into left anterior descending artery (LAD). We measured (1) coronary sinus level of ET-1, TNF-alpha using radioimmunoassay; (2) CFR, a measure of microvascular integrity, using Doppler flow wire in LAD at baseline and different doses of microspheres. RESULTS: CFR decrease significantly with different doses of microspheres (vs. baseline, P < 0.05). Level of ET-1, TNF alpha increased significantly with doses of 5 x 10(4) and peaked with 10 x 10(4). Interestingly, ET-1 progressively decrease while TNF-alpha persistently elevated from doses of 12 x 10(4) to 15 x 10(4). There are reverse correlations between ET 1 and CFR (r = -0.31, P < 0.05). CONCLUSIONS: The extent of microvascular injury wasn't linearly related to the extent of ME, where, it closely associated with myocardial ET-1. PMID- 17081372 TI - [C-reactive protein induced human endothelium cells apoptosis is associated with Bcl-2/Bax gene expression changes]. AB - OBJECTIVE: In the present study, we examined the expression changes of Bcl-2/Bax in C-reactive protein (CRP) treated human endothelium cells in vitro. METHODS: The human umbilical vein endothelial cells (HUVEC) were cultured by digest method for 2 - 3 posterities and incubated with human CRP (0, 1, 5, 25 mg/L for 24 hours) and analyzed by flow cytometer for apoptosis ratio. The effects of CRP in various concentrations on Bcl-2/Bax mRNA and protein expression were examined by RT-PCR and Western Blotting. RESULTS: Apoptosis ratio increased, downregulated Bcl-2 (gene promoting cell survival) and upregulated Bax (gene promoting apoptosis) at mRNA and protein levels in proportion to increased CRP concentrations. CONCLUSION: These results demonstrate that Bcl-2/Bax could be regulated by CRP in human HUVECs and might play a causal role in CRP-induced apoptosis. PMID- 17081373 TI - [siRNA inhibits efgl7 expression in human endothelial cell line HUVEC]. AB - OBJECTIVE: To evaluate the effects of small interference RNA (siRNA) on epidermal growth factor-like domain 7 (egfl7) gene expression in human endothelial cell line HUVEC. METHODS: siRNA targeting egfl7 (siRNA1, siRNA2, siRNA3 and siRNA4) was constructed through online design of Amnion company and transfected into human endothelial cell line HUVEC with lipofectamine. The nontransfected cells and cells treated with control siRNA were taken as controls. At 24, 48 and 72 hours post various interventions, cell viability was determined by MTS method as well as LDH and ATP releasing tests. egfl7 expressions at protein and mRNA levels were detected by Western blot and RT-PCR respectively. RESULTS: Cell survival rate, LDH and ATP release were significantly reduced in siRNA treated cells compared to control cells (P < 0.05). Similarly, egfl7 expression at protein and mRNA levels was also significantly reduced in siRNA treated cells (P < 0.01), especially in siRNA1 treated cells. CONCLUSION: siRNA inhibited egfl7 gene expression and cell survival in HUVEC. PMID- 17081374 TI - [Angiotensin II up-regulates expression of inducible nitric oxide synthase in human umbilical endothelial cells: roles of AT1 and AT2]. AB - OBJECTIVE: Angiotensin II is an important pro inflammation factor in the cardiovascular system. This experiment is aimed to study the effects of angiotensin II on inducible nitric oxide synthase expression in human umbilical endothelial cells. METHODS: Human umbilical endothelial cells were cultured in vitro and treated with angiotensin II alone or in combination with AT1, AT2 and NF-kappaB inhibitors respectively. The inducible nitric oxide synthase expressions at protein and mRNA levels were measured with Western blot and reverse transcription-polymerase chain reaction (RT-PCR), and the activity of NF kappaB was analyzed with EMSA. RESULTS: Angiotensin II up-regulated inducible nitric oxide synthase expressions at the protein and mRNA levels at 5 h (P < 0.05), the activity of NF-kappaB was enhanced at 2 h (P < 0.05). These effects could be blocked by AT1 and NF-kappaB inhibitors but not by AT2 inhibitor. CONCLUSION: Angiotensin II can upregulate the expression of inducible nitric oxide synthase through NF-kappaB pathway in human umbilical endothelial cells. AT1, other than AT2, play a key role in this process. PMID- 17081375 TI - [Major causes of death among male steelworkers in Beijing: a prospective study]. AB - OBJECTIVE: To evaluate the major causes of death and risk factors among male steelworkers in Beijing. METHODS: The study included 5137 men from the cohort of Beijing Capital Steel and Iron Company. The baseline survey was performed in 1974, 1979 and 1980 and the final follow-up evaluation was made in 2001 with a mean follow-up of 20.8 years. Causes of death were coded according to the Ninth Revision of International Classification of Diseases (ICD-9). The mortality was calculated by person-years of follow-up and age-standardized according to the 2000 census data in China. Cox proportional-hazards models adjusting for risk factors were used to estimate the relative risk of death. RESULTS: There is 760 deaths during follow-up. Age-standardized mortality from all causes was 643.0 per 100,000 person-years. The three leading causes of death were malignant neoplasms (mortality, 231.3 per 100,000 person-years), cerebrovascular diseases (mortality, 139.3 per 100,000 person-years) and heart diseases (mortality, 96.4 per 100,000 person-years). The multivariate-adjusted relative risk of death and the population attributable risk proportion for risk factors were as follow: cigarette smoking (95% CI, 1.174 to 1.765); hypertension (95% CI, 1.370 to 1.904) and hypercholesterolemia (95% CI, 1.057 to 1.537). CONCLUSIONS: Our study indicates that malignant neoplasms, cerebrovascular diseases and heart diseases were major causes of death among male steelworkers. Furthermore, cigarette smoking, hypertension and hypercholesterolemia are leading preventable risk factors for death. PMID- 17081381 TI - [Research progress of vascular endothelial progenitor cell]. PMID- 17081386 TI - [Summary of 9th National Conference on cardiovascular diseases by Chinese Medical Association]. PMID- 17081387 TI - [Protection against ischemia-reperfusion injury: from bench to bedside]. PMID- 17081388 TI - [Nitroglycerin tolerance aggravates arterial ischemia/reperfusion injury by increasing nitrotyrosine and ONOO- production]. AB - OBJECTIVE: To investigate the influence of nitroglycerin tolerance (NT) on arterial ischemia (90 min) and reperfusion (120 min). METHODS: Male Sprague Dawley rats were infused with nitroglycerin (GTN) or saline for 12 h and ascending aorta was rapidly isolated. The isolated aorta was subjected to one of the following treatments: stimulative ischemia/reperfusion, stimulative ischemia/reperfusion (I/R) plus glutathione (GSH, 0.1 mmol/L) during reperfusion, or control solution (Kreb's solution for 3.5 h). RESULTS: Compared with I/R group, contractile function, vasorelaxation responses to Ach, NO production were significantly decreased and CK, LDH activity as well as nitrotyrosine formation in reperfusion solution were significantly increased in I/R + NT group and these effects could be prevented with addition of GSH in I/R + NT aortas. CONCLUSIONS: Our results demonstrated that NT could aggravate arterial I/R injury by increasing the production of ONOO- and GSH may play a cardioprotective role against NT-induced myocardial injury by attenuating the formation of ONOO-. PMID- 17081389 TI - [Ischemic postconditioning attenuates ischemia/reperfusion injury in isolated hypertrophied rat heart]. AB - OBJECTIVE: To explore the effects of ischemic postconditioning on ischemia/reperfusion injury in isolated hypertrophied rat heart and investigate the signal transduction pathway changes induced by ischemia postconditioning. METHODS: Cardiac hypertrophy was induced in rats by abdominal aortic banding, and isolated hypertrophied rat heart ischemia/reperfusion model was made by Langendorff technique to evaluate the effects of ischemia postconditioning on left ventricular systole pressure, coronary artery flow, creatine phosphokinase (CPK) and lactate dehydrogenase (LDH) release, myocardial infarction size, and the level of myocardial phospho-protein kinase B/Akt (Ser473), phospho-glycogen synthase kinase-3beta (Ser9). Following groups were studied (n = 12 each group): IR, 30 min ischemia (I)/60 min Reperfusion (R); Post: 30 min ischemia, 6 circles of 10 s I/10 s R followed by 60 min R; Post Wort: 30 min ischemia, 6 circles of 10 s I/10 s R, wortmannin (10(-7) mol/L) followed by 60 min R; Wort: 30 min ischemia, wortmannin (10(-7) mol/L) followed by 60 min R. RESULTS: Left ventricular systolic pressure and coronary artery flow were significantly increased, myocardial infarction size and the release of CPK, LDH significantly reduced in Post group compared to that in IR group. Phospho-protein kinase B/Akt (Ser473) and phospho-glycogen synthase kinase-3beta (Ser9) levels were also significantly higher in Post group than that in IR group. Phosphatidylinositol 3 kinase (PI3K) inhibitor wortmannin prevented the increase of phospho-protein kinase B/Akt (Ser473) and phospho-glycogen synthase kinase-3beta (Ser9) induced by ischemic postconditioning, but only partly abolished the cardioprotection of ischemic postconditioning. CONCLUSION: Ischemic postconditioning attenuates ischemia/reperfusion injury in isolated hypertrophied rat heart. The cardioprotective effects of ischemic postconditioning were partly mediated through PI3K/Akt/GSK-3beta signaling pathway. PMID- 17081390 TI - [Role of gap junction in ischemic preconditioning]. AB - OBJECTIVE: To investigate the role of gap junction in ischemic preconditioning (IPC). METHODS: Sprague-Dawley rats were subjected to a 30 min coronary artery occlusion followed by 4 h of reperfusion (I/R). Rats were divided into seven groups: I/R, IPC/R, IPC/R + 5-hydroxydecanoic acid (mitochondrial ATP sensitive potassium channel antagonist), I/R + diazoxide (mitochondrial ATP sensitive potassium channel agonist), I/R + 5-hydroxydecanoic acid + diazoxide, I/R + 18beta-glycyrrhetinic acid (gap junction blocker) and I/R + 18beta-glycyrrhetinic acid + 5-hydroxydecanoic acid. Hemodynamics and myocardial infarct size were measured and connexin43 phosphorylation and subcellular distribution were determined by quantitative immunoblotting and confocal immunofluorescence. RESULTS: Infarct size was reduced in IPC/R, I/R + diazoxide and I/R + 18beta glycyrrhetinic acid group (13.34% +/- 7.87%, 11.02% +/- 2.24%, and 15.03% +/- 11.35%, respectively; P < 0.001 vs. I/R group: 45.81% +/- 7.91%). 5 hydroxydecanoic acid abolished the cardioprotective effects of IPC and diazoxide (46.57% +/- 5.36% and 47.36% +/- 3.17%; P > 0.05 vs. I/R) but not the effects of glycyrrhetinic acid (14.60% +/- 7.36%; P < 0.001 vs. I/R). Phosphorylation of connexin43 was significantly increased, dephosphorylation and connexin43 intracellular redistribution significantly decreased (Cx43 size in the cellular membrane 1.00% +/- 0.35% and 0.83% +/- 0.31%, P < 0.001 vs. I/R: 0.19% +/- 0.06%) by IPC and diazoxide and these effects could be abolished by 5-hydroxydecanoic acid. CONCLUSION: Ischemic preconditioning could reduce myocardial infarction size by activating mitochondrial ATP sensitive potassium channel and modulating connexin43 phosphorylation and internalization. PMID- 17081391 TI - [Comparable effects of percutaneous transluminal septal myocardial ablation and subaortic ventriculotomy in treating patients with hypertrophic obstructive cardiomyopathy, an echocardiography follow-up study]. AB - OBJECTIVE: To compare the effect of percutaneous transluminal septal myocardial ablation (n = 20) with subaortic ventriculotomy (n = 17) in patients with hypertrophic obstructive cardiomyopathy by echocardiography. METHODS: The pre- and post-operation (1 week post procedure) gradients of left ventricular out flow (LVOTGs) were measured by echocardiography. RESULTS: Post operation LVOTGs were significantly reduced than that of pre-operation in patients treated with either percutaneous transluminal septal myocardial ablation [(36 +/- 20) vs. (99 +/- 19) mm Hg (1 mm Hg = 0.133 kPa), P < 0.05], or subaortic ventriculotomy [(28 +/- 17) vs. (117 +/- 32) mm Hg]. Post operation LVOTGs were also similar with the two procedures [(36 +/- 20) vs. (28 +/- 17) mm Hg, P > 0.05]. CONCLUSION: Percutaneous transluminal septal myocardial ablation is as effective as subaortic ventriculotomy in treating patients with hypertrophic obstructive cardiomyopathy. PMID- 17081393 TI - [Familiar hypertrophic cardiomyopathy caused by a IVS15-1G > A mutation in cardiac myosin-binding protein C gene]. AB - OBJECTIVE: To detect the disease-causing gene mutation of hypertrophic cardiomyopathy (HCM) in a Chinese family and to analyze the correlation of the genotype and the phenotype. METHODS: One family affected with HCM was studied. The clinical data including symptom, physical examination, echocardiography and electrocardiography were collected. The full encoding exons and flanking sequences of beta-myosin heavy chain gene (MYH7) and cardiac myosin-binding protein C gene (MYBPC3) were amplified with PCR and the products were sequenced. RESULTS: A G8887A mutation, which is an acceptor splicing site of intron 15 (IVS15-1G > A) in MYBPC3 (gi: Y10129) was identified in 6 out of 11 family members. Three mutation carriers developed HCM at 48 - 75 years old with mild chest pain, chest distress and asymmetric septal hypertrophy (13 - 14 mm) and remaining mutation carriers are free of HCM. No mutation was identified in MYH7 gene. CONCLUSION: HCM caused by the IVS15-1G > A mutation is a benign phenotype. It is helpful to screen MYBPC3 gene mutation in late-onset HCM patients with mild symptoms. PMID- 17081394 TI - [IL-10 gene modification on immature dendritic cells induces antigen-specific tolerance in experimental autoimmune myocarditis]. AB - OBJECTIVE: To investigate whether IL-10 gene modification on immature dendritic cells (iDC) could induce autoimmune tolerance in rat experimental autoimmune myocarditis (EAM). METHODS: EAM was induced by cardiac myosin immunization on day 0 and day 7 in rats. A total of 2 x 10(6) mature DC (mDC), iDC, pcDNA3 transfected iDC, pcDNA3-IL-10 transfected iDC or PBS were injected intravenously at 5th immunization day. Three weeks later, echocardiography and HE staining were performed to observe the cardiac function and myocardial inflammation. Th1/Th2 cytokines were detected by ELISA and MHC-II molecules, costimulatory molecules were identified by flow cytometry. In vitro T lymphocyte proliferation assay and adoptive transfer of DCs were performed to determine the antigen specific tolerance induced by IL-10 gene modification on iDCs. RESULTS: EAM rats treated with pcDNA3-IL-10 transfected iDC showed improved cardiac function and reduced inflammatory cells infiltration into myocardium. Moreover, lower Th1 and higher Th2-type response was induced, MHC-II and costimulatory molecules down-regulated and antigen specific immunological responses towards cardiac myosin inhibited in pcDNA3-IL-10-iDC treated EAM rats. CONCLUSION: Treatment with IL-10 gene modified iDCs could ameliorates EAM by inducing Th2 polarization and down-regulation of MHC-II molecules and costimulatory molecule expressions. PMID- 17081396 TI - [Chronic effects of transmyocardial laser revascularization combined with off pump coronary artery by pass (OPCAB) compared with OPCAB alone in patients with ischemic heart disease: a prospective multicenter follow-up study]. AB - OBJECTIVE: To approach the long term safety and efficacy of transmyocardial laser revascularization (TMLR, holmium: YAG) combined with off-pump coronary artery bypass (OPCAB) compared with OPCAB alone in patients with ischemic cardiac disease. METHODS: Between 1999 and 2005, 80 patients with diffusely diseased target vessels from two centers in Beijing were enrolled to the study and randomized to receive either TMLR/OPCAB (n = 40) or OPCAB (n = 40) operation. Baseline demographics and operative characteristics were similar between groups. Follow-up (mean 3.4 +/- 1.7 years) included CCS angina class and NYHA classification assessments, 6 minutes walking test (6MWT) and echocardiography. RESULTS: Perioperative mortality was 5% in both groups. No death occurred during follow up. At the end of follow-up, patients at both groups experienced significant improvement on angina score compared with baseline, and angina score was also significantly lower (1.21 +/- 0.42 vs. 1.57 +/- 0.87, P = 0.03) and 6MWT distance significantly increased (518.0 +/- 65.5 m vs. 473.8 +/- 65.8m, P = 0.006) in OPCAB/TMLR group than that in the OPCAB group. Fewer patients developed recurrent severe angina and received re-CABG/PCI in OPCAB/TMLR group than that in the OPCAB (1 vs. 6 cases, P = 0.113). NYHA and LVEF were similar between the groups at the end of follow up. CONCLUSION: Our study showed that the addition of TMLR to OPCAB is superior in improving angina and exercise tolerance, but there is no further improvement in cardiac function compared to OPCAB alone. PMID- 17081397 TI - [Psychologic status comparison in patients treated with transradial or transfermoral approach coronary catheterizations]. AB - OBJECTIVE: We previously showed that factorial score of somatization, which was obtained by the examination of symptom checklist-90 (SCL-90), was higher in patients received transfemoral coronary catheterization than norm. The aim of the present study was to compare the patient's psychologic status between transradial approach and transfemoral approach percutaneous coronary catheterizations. METHODS: A total of 198 inpatients (105 transfemoral, 93 transradial) underwent scheduled first time coronary catheterizations were enrolled. All patients were studied by symptom SCL-90 on present psychologic status 24 hours before and 24-48 hours after coronary catheterizations. RESULTS: Age, sex, weight, smokers, employment, educational background, marriage status, family relations, family history of cardiovascular disease, income and medical insurance status were similar between the two groups. There was also no difference in diabetes, hypertension history as well as coronary heart disease confirmed by coronary catheterization between the 2 groups. Compared with the status before the procedure, factorial scores of somatization, obsessive-compulsive, interpersonal sensitivity, depression, anxiety, hostility, global severity index and total positive symptoms were significantly reduced after percutaneous coronary catheterizations (1.50 +/- 0.51 vs. 1.64 +/- 0.53, 1.50 +/- 0.48 vs. 1.67 +/- 0.55, 1.28 +/- 0.41 vs. 1.38 +/- 0.49, 1.42 +/- 0.43 vs. 1.55 +/- 0.53, 1.38 +/- 0.41 vs. 1.58 +/- 0.54, 1.32 +/- 0.35 vs. 1.44 +/- 0.41, 1.38 +/- 0.34 vs. 1.49 +/- 0.42, and 23.08 +/- 17.30 vs. 27.72 +/- 18.79, respectively, P all < 0.05). Scores on somatization, depression and positive symptom severity index were significantly lower in patients received transradial coronary catheterizations than those received transfemoral coronary catheterization approach (1.52 +/- 0.51 vs. 1.62 +/- 0.53, 1.43 +/- 0.54 vs. 1.54 +/- 0.43 and 2.36 +/- 0.66 vs. 2.50 +/- 0.43, respectively, P all < 0.05). CONCLUSION: Patients' psychologic status improved significantly after percutaneous coronary catheterizations. Improvement on psychologic status is significantly better in patients underwent transradial coronary catheterizations than that underwent transfemoral coronary catheterizations. PMID- 17081398 TI - [Predictive value of fibrinogen and high-sensitivity C-reaction protein for cardiovascular events in patients with stable coronary artery disease]. AB - OBJECTIVE: To investigate the predictive value of elevated fibrinogen and high sensitivity C-reaction protein (hs-CRP) level on cardiovascular events in patients with stable coronary artery disease (CAD). METHODS: From January 2002 to November 2002, 185 patients (aged 47 - 85 years) with stable CAD referred for coronary angiography were enrolled and divided into control-F (fibrinogen level < or = 4.0 g/L, n = 104) and elevated-F (fibrinogen level > 4.0 g/L, n = 81), or control-hs (hs-CRP < or = 3.0 mg/L, n = 99) and elevated-hs (hs-CRP> 3.0 mg/L, n = 86). Exclusion criteria included cardiomyopathy, New York Heart Association class IV congestive heart failure, recent myocardial infarction or coronary artery revascularization and cancer. During three years follow-up, cardiovascular death, myocardial infarction, congestive heart failure, stroke and other vascular events were assessed. RESULTS: A total of 21 cardiovascular nonfatal events and 10 cardiovascular deaths were observed. Cardiovascular events was significantly higher in patients in elevated-F group than that in control-F group [23.46% vs. 11.54%, cholesterol-, body mass index-, smoking-, and hypertension-adjusted relative risk 1.97, 95% CI (1.68 to 2.40), P < 0.05] and in elevated-hs group than in control-hs group [24.42% vs. 10.10%, adjusted relative risk 2.32, 95% CI (1.76 to 2.89), P < 0.05]. The relative risk of cardiovascular events for patients with fibrinogen > 4.0 g/L and hs-CRP > 3.0 mg/L was 3.84 (P < 0.05), 95% CI (2.80 to 4.99) compared with patients with fibrinogen < or = 4.0 g/L and hs CRP < or = 3.0 mg/L. CONCLUSION: Both fibrinogen and hs-CRP are independent important predictors of cardiovascular nonfatal and fatal events in patients with stable CAD. Combination of elevated fibrinogen and hs-CRP increased their predictive value for cardiac events. PMID- 17081399 TI - [Atherosclerotic aortic ulcers monitoring by electron beam CT or multi-slice CT]. AB - OBJECTIVE: To evaluate the evolution of medically treated atherosclerotic aortic ulcers by computed tomography (CT). METHODS: Thirty-five patients (31 men and 4 women, aged from 40 to 79 years, mean 56.2 +/- 10.8 years) with known aortic ulcers were monitored by CT (follow up time 7 - 730 days, mean 135 days), 80 - 100 ml contrast media (Ultravist 300 or 320, or Omnipaque 300 or 320 mg/ml) was injected with a rate of 3.5 - 4.5 ml/s. The scan delayed time was 18 - 30 s. Ulcers dimensions were measured according to maximum depth, maximum length and maximum width. RESULTS: Thirty-one patients with intramural hematomas and 1 patient with atherosclerotic aortic arch aneurysm without intramural hematoma were medically treated and another 3 patients were surgically treated. Intramural hematoma regression was monitored in 31 medically treated patients with intramural hematomas. CT was repeated at 2 weeks, 3 and 6 months. Intramural hematoma resolved gradually during follow up [thickness: (7.69 +/- 4.24) mm at 3 months, (3.06 +/- 1.67) mm at 6 months, P < 0.05 vs. 1st CT: (11.96 +/- 4.16) mm while ulcer maximum depth (11.17 +/- 6.03) mm at 3 months, (11.35 +/- 5.59) mm at 6 months, P < 0.05 vs. 1st CT: (7.36 +/- 6.61) mm, maximum width (14.40 +/- 6.35) mm at 3 months, (18.55 +/- 10.94) mm at 6 months, P < 0.05 vs. 1st CT: (7.15 +/- 6.39) mm, maximum length (17.12 +/- 7.15) mm at 3 months, (18.13 +/- 10.89) mm at 6 months, P < 0.05 vs. 1st CT: (11.64 +/- 10.06) mm increased progressively during follow-up]. CONCLUSION: CT was a useful tool for deflecting atherosclerotic aortic ulcers and monitoring therapeutic effects. PMID- 17081400 TI - [The effects of phytoestrogen on blood pressure and lipids in healthy volunteers]. AB - OBJECTIVE: To determine the effects of dietary soy containing phytoestrogens on blood pressure and lipids in healthy volunteers. METHODS: Two hundred thirteen healthy volunteers (108 men and 105 post-menopausal women, 50 - 76 years old) received either soy protein isolate (40 g soy protein, 118 mg isoflavones) or cassin placebo for 3 months in this randomized, double-blind trial. RESULTS: There were 34 withdrawals (16%) and 179 people (96 men and 83 women) completed the study protocol. After 3 months treatment, urinary phytoestrogens was significantly increased and blood pressure was significant reduced in soy protein group than that in cassin placebo group [mean change in systolic (-7.5 +/- 1.2) mm Hg vs. (-3.6 +/- 1.1) mm Hg, P < 0.05; diastolic: -4.3 +/- 0.8) mm Hg vs (-1.9 +/- 0.7) mm Hg, P < 0.05; mean aortic blood pressure: (-5.5 +/- 1.0) mm Hg vs ( 0.9 +/- 1.0) mm Hg, P < 0.008]. Low- to high-density lipoprotein ratio [(-0.33 +/ 0.10) mmol/L vs (0.04 +/- 0.10) mmol/L, P < 0.05] and triglycerides [(-0.20 +/- 0.05) mmol/L vs (-0.01 +/- 0.05) mmol/L, P < 0.05] were significantly reduced and Lp(a) lipoprotein significantly increased [42 (17 - 67) mg/L vs 4 (22 - 31) mg/L, P < 0.05] in soy protein group compared to cassin placebo group. Total, low density lipoprotein, and high-density lipoprotein cholesterols all improved in both groups and were similar between the groups. No side-effect was observed in both groups and no effect on the hypothalamic-pituitary-gonadal axis was noted in study subjects. CONCLUSION: In normotensive men and post-menopausal women, phytoestrogens intake improved blood pressure and lipids status. PMID- 17081402 TI - [Effect of cardiotrophin-1 on cardiac transcription factor GATA4 expression in rat cardiomyocytes]. AB - OBJECTIVE: To investigate the effect of cardiotrophin-1 (CT-1) on the GATA4 expression and related signaling pathways (JAK-STAT3, ERK1/2 and PI3-K) in rat cardiomyocytes. METHODS: Using semi-quantitative RT-PCR and EMSA, we measured the dose and time dependent effects of CT-1 on GATA4 mRNA and binding activity in cultured rat cardiomyocytes. Parthenolide (a STAT inhibitor), U-0126 (an ERK inhibitor) and LY-294002 (a PI3-K inhibitor) alone or in combination were added to the culture medium to assess the role of above signaling pathways in CT-1 mediated effects. RESULTS: GATA4 mRNA expression significantly increased at 3 h post 0.1 nmol/L CT-1 exposure, peaked at 6 h and remained high till 24 h post exposure. The GATA4 binding activity began to increase at 10 min and peaked at 60 min and returned to baseline level 180 min. Six hours post CT-1 (0.01 nmol/L, 0.1 nmol/L, 1 nmol/L) exposure, the GATA4 mRNA expression increased in a dose dependent manner. The GATA4 binding activity peaked with 0.1 nmol/L CT-1 and higher dose did not further increase the binding activity. U-0126 increased the GATA4 mRNA expression and enhanced the GATA4 binding activity and these effects could be partially attenuated with addition of Parthenolide. Parthenolide also prevented the increase of GATA4 mRNA and binding activity induced by CT-1. LY 294002 had no effects GATA4 mRNA and binding activity. CONCLUSION: CT-1 increases the GATA4 mRNA expression and binding activity in rat cardiomyocytes via STAT3/ERK1/2 pathways and these effects are independent of PI3-K pathway. PMID- 17081403 TI - [Association between calcium-sensing receptor protein expression and rat cardiomyocyte apoptosis]. AB - OBJECTIVE: To investigate the relationship between calcium-sensing receptor protein (CaSR) expression and rat cardiomyocyte apoptosis and related signal transduction pathways. METHODS: The CaSR, BCl2, Caspase3 protein and ERK1/2 phosphorylation or non-phosphorylation were detected by Western blot. Cardiomyocyte apoptosis was detected by flow cytometry and immunofluorescence. RESULTS: CaSR protein was detected in rat cardiac tissue and CaSR activator gadolinium (GdCl3) induced cardiomyocyte apoptosis and increased ERK1/2 phosphorylation and expression of BCl2 and activated Caspase3. The selective mitogen-activated protein kinase (MAPK) inhibitor PD98059 abolished gadolinium induced ERK1/2 activation and BCl2 expression, further increased the activation of Caspase3 and cardiomyocyte apoptosis. CONCLUSION: Our results demonstrate the CaSR existence in cardiomyocytes and CaSR activation by gadolinium can induce myocyte apoptosis by activating Caspase3 and tyrosine protein kinase pathway. PMID- 17081404 TI - [Cardiac valve MCSF-1 and its receptor expression changes in Staphylococcus aureus induced infective endocarditis]. AB - OBJECTIVE: To explore the cellular immunology mechanism of infective endocarditis (IE), we investigated the effects of Staphylococcus aureus (S. aureus) on MCSF-1 and its receptor (c-fms) gene expression in cardiac valves. METHODS: Thirty-two rabbits were divided into 4 groups: mitral or tricuspid valve artificial lesions with 5 x 10(4) CFU or 5 x 10(6) CFU S. aureus injection. Control rabbits (n = 7) received 5 x 10(6) CFU S. aureus injection. IE after operation were confirmed by naked eyes and electron microscope observations. MCSF-1, c-fms in mitral and tricuspid valves were detected by RT-PCR. RESULTS: Twenty-six rabbits survived the operation and 14 rabbits developed IE (2 with 5 x 10(4) CFU and 12 with 5 x 10(6) CFU S. aureus injection) one day post operation. S. aureus injection alone did not induce IE. Compared to control rabbits, MCSF-1 mRNA was significantly upregulated and c-fms mRNA significantly downregulated after 5 x 10(4) CFU S. aureus injection with heart valve artificial lesion in mitral valves or tricuspid valves. MCSF-1 expression in mitral valves was further increased while remained unchanged in tricuspid valve after 5 x 10(6) CFU S. aureus injection compared to that in 5 x 10(4) CFU S. aureus injection group. CONCLUSION: High dose bacterial invasion and heart valves lesion were the main factors for inducing infective endocarditis. Development of infective endocarditis was associated with valve MCSF-1/c-fms expression changes in this rabbit model. PMID- 17081410 TI - [The advance of myocardial reperfusion on tissue level]. PMID- 17081405 TI - [A cohort study on parental history of hypertension and the risk of cardiovascular disease in Chinese population]. AB - OBJECTIVE: To assess the relationship between parental history of hypertension and the incidence of cardiovascular disease in Chinese populations. METHODS: A total of 15 131 individuals (7015 men and 8116 women, aged from 35 to 59 years) in China were enrolled and followed up from 1992 to 2005. The information on the parental history of hypertension of the participants was collected. The individuals were divided into three groups according to their parental history of hypertension: those without parental history of hypertension (group 1), those with one parent history of hypertension (group 2) and those with both parents history of hypertension (group 3). The hazard ratio and its 95% confidence interval were calculated using the Cox proportional hazard model. RESULTS: There were 448 cardiovascular events (including 82 cardiac events and 370 stroke events and 4 with both cardiac and stroke events) during the 163 858 person-years of observation. After adjusting for age, smoking and drinking, the hazard ratio for cardiovascular disease for group 1, group 2 and group 3 were 1.00, 1.34 (1.01 - 1.78), 2.58 (1.62 - 4.11) in men respectively and 1.00, 1.77 (1.27 - 2.45), 2.55 (1.44 - 4.54) in women respectively. After further adjusting for serum total cholesterol (TC), high density lipid cholesterol (HDL-C), fasting glucose, body mass index (BMI) and systolic blood pressure (SBP) in the model, the hazard ratio for cardiovascular disease for those three groups were 1.00, 1.01 (0.76 - 1.35), 1.72 (1.07 - 2.75) in men respectively and 1.00, 1.31 (0.94 - 1.84), 1.76 (0.98 - 3.15) in women respectively. CONCLUSION: The individuals with parental history of hypertension have a higher risk of cardiovascular disease, especially for those with both parents history of hypertension. PMID- 17081412 TI - [Comprehensive therapy of orbital tumor]. AB - Although the popularization of the knowledge of orbital disease is increasing, some issues such as surgery complications, recurrence, metastases may significantly affect the outcome of treatment. The new concept of comprehensive therapy offers an integrated individual therapy to patients. Application the concept in clinical practice is a key measure to improve the cured rate. In order to enhance the rate of cure of orbit tumor, it is essential for ophthalmologist to get standardized training in the diagnosis, treatment of orbital disease, and familiar with orbital anatomy and other knowledge related to orbit diseases. PMID- 17081413 TI - [Clinical and pathological analysis of choroidal metastatic carcinoma]. AB - OBJECTIVE: To study the clinical and pathological characteristics of choroidal metastatic carcinoma. METHODS: The clinical data, pathological character, primary tumor origin and histological classification of 18 patients with choroidal metastatic carcinoma were analyzed retrospectively. RESULTS: Most patients had severe visual impairment. Solid mass was seen in the posterior pole of the eyes in all 18 patients through ocular fundus examination, 8 cases had retinal detachment. B scan and CT examination found flat or irregular masses. MRI examination had been performed on 5 patients, high signal intensities on T1W and low signal intensities on T2W were found. Five patients were adenocarcinoma, 4 were squamous carcinoma and 3 were undifferentiated carcinoma through pathological examination. Primary tumor was lung carcinoma in 10 cases (55%) and breast carcinoma in 4 cases (22%). CONCLUSIONS: Rapid decrease of visual acuity, flat neoplasm in ocular fundus and secondary retinal detachment are the main clinical characteristics of choroidal metastatic carcinoma. The most common primary tumor is lung carcinoma and the most common histopathological classification is adenocarcinoma. Imaging examination is helpful for the diagnosis of choroidal metastatic carcinoma. PMID- 17081414 TI - [Retrospectively analysis of diagnosis and treatment of the orbital neurilemomas]. AB - OBJECTIVE: To evaluate the diagnosis and surgery of orbital neurilemoma. METHODS: Thirty-eight patients with pathologically confirmed orbital neurilemomas were analyzed retrospectively. The age of the patients ranged from 9 years to 69 years. Twenty-three patients were female and fifteen were male. Standardized A/B scan was performed in 37 cases. CT scanning was performed in 38 patients with contrast scanning in 15 patients and MR imaging was performed in 33 patients with contrast scanning in 32 patients. RESULTS: Neurilemoma was generally low to medium reflective or echo in the ultrasound. Of the 38 cases with orbital neurilemomas, location of tumors was superior of the orbit in 17 cases; lateral and inferior in 19 cases and medial in 2 cases. Shape of the tumors was elongate or round-like 26 cases, leaf-like and dumb-bell 12 cases. CT scanning showed soft tissue high density mass in 32 cases, 6 cases were heterogeneity (for cystoid change). MR imaging in 33 patients displayed a identical T1 and a long T2 signal intensity weighted images in 4 cases; identical T1 and T2 mixed signal (short, identical and long signal) in 28 cases; identical T1 and T2 in 1 cases. After Gd DTPA enhancement in 32 cases, heterogeneity enhancement of the mass was seen in 28 cases, marked enhancement 4 cases. Four cases in all cases was found orbital cranial communicate mass. Surgical approach was lateral orbitotomy in 26 cases, combined lateral and medial orbitotomy in 2 cases, anterior orbitotomy 4 cases and medial orbitotomy in 2 cases. CONCLUSIONS: Modern medical imaging, especial MR imaging can demonstrate the site, configuration, and characteristics of the tumors and contribute to the diagnosis and treatment. Skill of surgery can improve the rate of success and decrease the complications. PMID- 17081415 TI - [Analysis of the relationship between pathology and recurrence of primary lacrimal epithelial tumors]. AB - OBJECTIVE: To analyze the relationship between pathology and recurrence of primary lacrimal epithelial tumors. METHODS: 128 cases of primary lacrimal epithelial tumors including benign mixed tumor (74 cases, 57.8%), adenoid cystic carcinoma (22 cases, 17.2%) and malignant mixed tumor (18 cases, 14.1%) were subjected in the study. Pathological features were analyzed and compared with their recurrence. RESULTS: The recurrent rate of benign mixed tumor, adenoid cystic carcinoma and malignant mixed tumor was 23.0%, 18.2% and 27.8%, respectively. The recurrence of benign mixed tumor was statistically related to pathological classification and encapsulates. CONCLUSIONS: Primary lacrimal epithelial tumors show variant types and high recurrent rate. The pathological features were decisive in diagnosis, selection of treatment and the time of follow-up. PMID- 17081416 TI - [The effects of matrine on cell proliferation and telomerase activity in retinoblastoma cells in vitro]. AB - OBJECTIVE: To evaluate the effects of matrine on cell proliferation and telomerase activity in retinoblastoma cells in vitro. METHODS: HXO-Rb44 cells were treated with 0.1 to 3.2 mmol/L matrine for 24 h, then cell viability was measured by MTT assay. Apoptosis rates were evaluated by flow cytometry and TUNEL assay after the cells were treated for various durations. The telomerase activity were measured by telomeric repeat amplification protocol. RESULTS: MA inhibited the growth of HXO-Rb44 cells. Ma induced the apoptosis and down-regulated the telomerase activity of HXO-Rb44 cells, which were time dependent. There was a positive correlation between the telomerase activity and the apoptosis (r = 0.961, P < 0.01). CONCLUSIONS: Ma can induce the apoptosis and down-regulate the telomerase activity of retinoblastoma cells, which may be one of important mechanisms to inhibit the cell proliferation. It suggests that Ma may be a potent drug for the treatment of retinoblastoma. PMID- 17081417 TI - [Vitrectomy for severe proliferative diabetic retinopathy: a comparison of optical fiber free intravitreal surgery system with traditional vitrectomy]. AB - OBJECTIVE: To compare the optical fiber free intravitreal surgery system with traditional vitrectomy in eyes with severe proliferative diabetic retinopathy (PDR). METHODS: 22 eyes of 18 patients with extensive fibrovascular proliferation associated with PDR were undergone vitrectomy using optical fiber free intravitreal surgery system (OFFISS) with bimanual technique, peeling of epiretinal membrane and/or internal limiting membrane, intraocular laser coagulation, air-fluid exchange, tamponading of air and expansive gases or silicon oil were performed according to the condition of diseases. 22 eyes with similar severity of fibrovascular proliferation treated with traditional vitrectomy were served as control. RESULTS: The operations were finished successfully in both different vitrectomy systems, but fundus cannot be visualized during fluid-gas exchange in phakic eyes, and intraocular illumination had to been used again in OFFISS group. Postoperatively, retina was fully attached in all eyes, best-corrected visual acuity of 19 eyes had improved two lines and more in OFFISS group, and 17 eyes in control group. No significant difference in surgical complications was found between OFFISS and traditional vitrectomy group. Mean duration of peeling membrane was less in OFFISS group than control group. CONCLUSIONS: The efficacy of vitrectomy using different system for severe PDR both are satisfactory and comparable. We consider vitrectomy using OFFISS more safe and effective during complicated surgical manipulations than using traditional vitrectomy system. PMID- 17081418 TI - [The effect of retrobulbar injection with anisodamine on ocular blood velocity in anterior ischemic optic neuropathy]. AB - OBJECTIVE: To study the effect of retrobulbar injection with anisodamine on ocular blood velocity in anterior ischemic optic neuropathy (AION). METHODS: 15 minutes before injection, 15 minutes and 1 hour after injection, the blood flow velocity in 39 cases (39 eyes) diagnosed as AION was measured and analyzed by CDI, timed average maximum velocity, peak systolic velocity, end diastolic velocity, resistance index and pulsatility index of OA, CRA and PCA were recorded. RESULTS: Compared with the normal eyes, each parameter of SPCA was statistically significant. 15 minutes and 1 hour after post ocular injection of anisodamine, every parameter examined in the experiment was increased except resistance index compared with pre-injection (P < 0.01). CONCLUSIONS: retrobulbar injection with anisodamine can effectively improve flow velocity of nasal and tempo short posterior ciliary artery and central retinal artery. PMID- 17081419 TI - [Late management of medial canthal deformities after naso-orbital-ethmoid fractures]. AB - OBJECTIVE: To evaluate the surgery methods and effectiveness of medial canthal deformities after naso-orbital-ethmoid fractures (NOE) and analysis the long-term results. METHODS: The clinical data and following-up results from 83 patients (83 eyes), which were treated in the Department of Ophthalmology, Ninth People's Hospital, Medical School of Shanghai Jiao Tong University from January 1999 to January 2004, were retrospectively analyzed. All the patients had unilateral medial canthal deormities after NOE fractures, and they were on operations of medial canthoplasty and fixation of the canthal ligament. Based on the severity of the canthal deformities and local scar of the area, different skin flaps were utilized for canthoplasty, such as "Y-V" flap, "Z" flap, "Mustard" method and rotation flaps. According to the fracture pattern and severity of the medial canthal tendon's injury, different medial canthopexy were performed such as suturation, fixation with titanium nail, drilling a role across the bone, and reposition the "central segment" using titanium miniplates. The surgery to repair the obstruction of the lacrimal system were accompanied with canthoplasty at the same time, while the orbital surgery for repositon of the enophthalmos was often performed at the first stage, and then the deformities of medial canthus were corrected after more than six months. The follow-up time varied from 12 to 30 months. RESULTS: 72 cases obtained satisfied contour of the medial canthal area, the position of the canthus was symmetry with the normal side, especially the formation of the canthal fossa. Recurrence of deformities occurred in 11 cases in six months after canthoplasty, among them 6 patients were re-operated for the second time. CONCLUSION: According to the fracture pattern and severity of the medial canthal deformity, different operative designs and canthoplasy can produce satisfied results. PMID- 17081420 TI - [The autonomic pupillary dysfunction in type II diabetes mellitus]. AB - OBJECTIVE: To evaluate the pupillary disorder associated with autonomic neuropathy of type II diabetes mellitus by investigating pupil diameters under mesopic, photopic and pharmacologically dilated conditions. METHODS: Forty of type II diabetic patients were divided into two groups based on the results of fundus fluorescein angiography, one group was the subclinic diabetic retinopathy and another was the NPDR (nonproliferative diabetic retinopathy), 20 age-matched healthy subjects were selected as control. High-resolution images of the pupil in 60 subjects were taken using an infrared-sensitive camera under mesopic, photopic and pharmacologically dilated conditions, respectively. From the images, the pupil diameters, constriction ratio and dilatation ratio were analyzed using Photoshop, Acdsee, and Imagetools software. RESULTS: The mean pupil diameter of tested subjects in mesopic control, subclinic and the NPDR were (6.02 +/- 0.48), (5.87 +/- 0.99), (4.95 +/- 1.12)mm, respectively. The mean photopic pupil diameter in three groups were (3.40 +/- 0.33), (3.37 +/- 0.31), (3.25 +/- 0.47) mm, respectively, where the mean pharmacologically dilated pupil diameter in three groups were (7.37 +/- 0.59), (6.91 +/- 1.00), (5.49 +/- 1.24) mm, respectively. The mean constriction ratio of these three groups were (43 +/- 7)%, (41 +/- 10)%, (32 +/- 14)%, respectively. The mean dilatation ratio of these three groups were (23 +/- 8)%, (19 +/- 13)%, (11 +/- 5)%, respectively. There were statistically significant differences among three groups in dark adapted pupil diameter, dilated pupil diameter, constriction ratio and dilatation ratio (P < 0.05). CONCLUSIONS: Autonomic pupillary dysfunction occurs early in type II diabetic patients. Pupillary diameters under mesopic and pharmacologically dilated conditions, and other pupil changes such as constriction ratio and dilatation ratio can be the reliable sign to evaluate autonomic neuropathy of type II diabetes mellitus. PMID- 17081421 TI - [Oblique nuclear chopping and nuclear extraction by binding procedure for cataract extraction]. AB - OBJECTIVE: To investigate the effects of oblique nuclear chopping and nuclear extraction by binding procedure (ONCNEB) in cataract extraction. METHODS: Extracapsular cataract extraction by ONCNEB with foldable IOL implantation was performed in 86 senile cataract cases (97 eyes) by using a line-shaped cataract nuclear chopping apparatus developed by the authors. RESULTS: In 97 eyes treated with ONCNEB, there was posterior capsular rupture in 1 eye; anterior chamber hemorrhage in 2 eyes; elevated intraocular pressure in 6 eyes; mild corneal edema in 8 eyes which subsided within 5 days. Corrected visual acuity > or = 0.5 was obtained in 78 eyes (80.41%) one week and 91 eyes (93.81%) three months after the operation. Corrected visual acuity was > or = 1.0 in 66 eyes (68.04%) three months after the operation. Mean astigmatism one week and three months postoperatively showed not significant difference as compared with preoperative astigmatism. CONCLUSION: The ONCNEB can solve the problem of implanting foldable IOL through a small incision. This procedure is relatively simple and safe. PMID- 17081422 TI - [The effects of amniotic membrane on corneal penetration of ofloxacin]. AB - OBJECTIVE: To evaluate the corneal penetration of ofloxacin following amniotic membrane transplantation (AMT). METHODS: One hundred and eight New Zealand rabbits were divided into 6 groups: mechanical remove of corneal epithelium with and without AMT; corneal alkali burn with and without AMT, and normal cornea with and without AMT. Topical ofloxacin was administered four times at interval of every 15 minutes. Five min, 30 min, 1 h, 2 h, 4 h and 6 h after the last administration, 3 rabbits from each group were sacrificed and aqueous humor was collected. The concentration of ofloxacin was detected by high pressure liquid chromatography (HPLC). RESULTS: The ability of ofloxacin to penetrating cornea was improved obviously in de-epithelialized groups and cornea alkali burn groups no matter AMT was performed or not (P < 0.05). The concentration of ofloxacin in the aqueous humor in normal cornea with and without AMT showed no statistical difference within 1 h after administration. But at the last three time points, the ofloxacin concentration in AMT group was greater than that in the group without AMT (P < 0.05). The comparison between the de-epithelialized with and without AMT obtained the similar result. At every tested time point, the ofloxacin concentration in aqueous humor of corneal alkali burn with AMT group was statistically greater than that of the group without AMT (P < 0.05). CONCLUSION: Amniotic membrane could store and release the drug slowly, results in the increase of contact time between ofloxacin and corneal surface and promotes the corneal penetration of ofloxacin. PMID- 17081423 TI - [Aspergillus fumigatus activate human corneal epithelial cells via Toll-like receptor 2 and 4]. AB - OBJECTIVE: To determine Toll-like receptors (TLR) 2 and TLR4 expression in human corneal epithelial tissue and cell line (THCE), and its activation by aspergillus fumigatus (AF) in inflammatory response. METHODS: The expression of TLR2 and TLR4 protein in human corneal epithelial tissue and THCE was detected by Western blot and immunocytochemistry. THCE was challenged with AF mycelium fragment (5 x 10(6)/ml) and supernatant extract agent (equivalent to bovine serum albumin 10 microg/ml). IL-8 and TNF-alpha in THCE supernatant were detected by ELISA at 1, 2, 4 and 8 h post stimulation. The protein of IkappaBalpha in THCE cells was assayed by Western blot at 30 min, 1 h and 2 h after treatment. Antibody blocking test was utilized to evaluate the effect on IL-8 and TNF-alpha expression of THCE by blocking TLR2 and (or) TLR4 before challenge with AF agent. RESULTS: TLR2 and TLR4 protein were expressed in human corneal epithelial tissue and THCE. The IL-8 and TNF-alpha level in THCE supernatant was elevated at 1 h, increased to (64.71 +/- 5.15) pg/ml and (32.46 +/- 3.28) pg/ml (AF mycelium challenge group), (94.94 +/- 11.92) pg/ml and (48.70 +/- 3.32) pg/ml (AF supernatant challenge group) 8 h post-challenged, which was 3.0 times and 2.5 times, 4.5 times and 3.5 times to that of control group respectively (P < 0.01). The activity of IkappaBalpha in THCE cells was decreased to 10.31 +/- 1.30 (gray scale value) and 8.15 +/- 2.37 at 30 min after challenged with AF mycelium or supernatant extract agent compared to 51.57 +/- 5.58 and 49.23 +/- 3.49 of control group (P < 0.01), and was reverted at 2 h. The secretion of IL-8 and TNF-alpha was partly inhibited by blocking TLR2 or TLR4 (P < 0.05), obviously inhibited by blocking TLR2 and TLR4 (50% and 40% compared to that of control group) (P < 0.01) when challenged with AF mycelium. And that was markedly inhibited by blocking TLR4 or blocking TLR2 and TLR4 when challenged with AF supernatant (P < 0.01). The secretion of IL-8 and TNF-alpha was not inhibited by blocking TLR2 when challenged with AF supernatant (P > 0.05). CONCLUSIONS: AF agent may induce human corneal epithelial cells express inflammatory cytokines via TLR-NF-kappaB pathway. TLR2 and TLR4 possibly mediate the recognition to AF mycelium, and TLR4 may dominate the recognition to AF supernatant agent. PMID- 17081424 TI - [Fluorescein angiography of retinal neovascularization model in mice]. AB - OBJECTIVE: To investigate a high molecular weight fluorescein angiography in the application of retinal neovascularization model in mouse. METHODS: retinal neovascularization model was induced by exposure mouse to an environment containing high concentration of oxygen. High molecular weight fluorescein isothiocyanate dextran were perfused through the left ventricle directly, then the mouse eyes were enucleated and fixed with 4% paraformaldehyde. The retina was separated from the eyecup and flat mounting was performed on a gelatin coated slide. The vasculature was examined under fluorescent microscope. RESULT: The whole retinal vasculature was clearly visualized under fluorescent microscope. By focusing on different layer of the tissue, superficial, deep vascular plexus and connecting vessels also could be distinguished. The neovascular response occurred at the junction between the vascular and the avascular retina. CONCLUSION: High molecular weight fluorescein angiography can be applied for retinal neovascularization evaluation. PMID- 17081425 TI - [Ocular pharmacokinetics and bioavailability of 0.2% ganciclovir in-situ gelling eye drops]. AB - OBJECTIVE: To compared the pharmacokinetics and bioavailability of 0.2% ganciclovir (GCV) in-situ gelling eye drops (GCV-ISG) with common GCV eye drops (GCV-ED) in rabbits. METHODS: Forty-eight healthy albino rabbits were randomly divided into 2 groups, each group included 24 rabbits and 3 rabbits (6 eyes) were used at each time points. Each eye received 50 microl of GCV-ISG in experimental group, and the same amount of GCV-ED was given in the other group as the control. The tears and aqueous humors were collected at 5, 10, 15, 30, 45, 60, 90 and 120 minutes following topical application of GCV-ISG and GCV-ED, respectively, and the corneas were immediately dissected after euthanized. The samples from 6 eyes (3 animals) were obtained at each designed time point. All samples were stored 60 degrees C and then were assayed by reversed phase high performance liquid chromatography (HPLC). An unpaired Student's t-test and 3p97 pharmacokinetics software were used as statistical analysis. RESULTS: The drug levels in tears corneas and aqueous were significantly higher for GCV-ISG group than GCV-ED group at 5, 10 minutes (P < 0.05) and 120 minutes (P < 0.01). The areas under the curve (AUC0-120) of the drug concentrations versus times in tears, corneas and aqueous humors for GCV-ISG group were 2.22, 5.47 and 3.40 times as high as GCV-ED group within designed duration, respectively. The peak concentrations of GCV in aqueous humors for GCV-ISG group and the GCV-ED group were 4.79 microg/ml and 0.96 microg/ml, respectively. The half-lives of GCV in aqueous humors and corneas for GCV-ISG group were 59 minutes and 223 minutes, and for GCV-ED group were 43 minutes and 87 minutes, respectively. The peak concentration of GCV in aqueous humor in GCV-ISG group was 4.99 times higher than that in GCV-ED group (P < 0.01). CONCLUSIONS: 0.2% ganciclovir in-situ gelling eye drops significantly increases the drug penetration into cornea and aqueous humor, and prolongs the residence time in cornea and aqueous humor. The results suggest that 0.2% ganciclovir in-situ gelling eye drops may enhance the ocular bioavailability of ganciclovir in rabbit eye. PMID- 17081426 TI - [Establishing the linkage disequilibrium pattern for the all-trans-retinol dehydrogenase (RDH8) gene]. AB - OBJECTIVE: As part of an on-going effort to map genes involved in complex eye diseases, myopia in particular, single nucleotide polymorphisms (SNPs) and linkage disequilibrium (LD) pattern were used to identified the gene within and around the All-trans-retinol dehydrogenase (RDH8). METHODS: Denaturing high performance liquid chromatography (DHPLC) was used to screen SNPs in 4 DNA pools each consisting of DNA from five individuals, and genotypes identified SNPs coupled with DNA pooling strategy were performed in 150 Chinese subjects from Hong Kong. The identified common SNPs were included in LD and haplotype analysis using the Haploview2.05 and EH programs. RESULTS: Fifteen SNPs were identified: 7 were common with the minor allele frequency > 0.05, and 10 were novel. Four SNPs in the 3' region exhibited significant LD (/|D'/ > 0.75 and its confidence interval suggesting strong LD, r2 > 0.33, P < 0.031) and formed a haplotype block while 3 common SNPs in the 5' region did not exhibit obvious LD. CONCLUSION: The block-like LD pattern existed around the RDH8 gene region suggest that one SNP (RDH8E5a probably) in the 3' region and at least 2 SNPs in the 5' region (RDH851 particularly) were needed in association studies involving RDH8. PMID- 17081427 TI - [Apoptosis of lens epithelial cell induced by curcumin and its mechanism]. AB - OBJECTIVE: To investigate the effects of natural drug curcumin (Cur) on apoptosis of lens epithelial cell (LEC) in vitro and its mechanism. METHODS: The bovine LEC were cultured with Cur, the ultrastructure changes were observed under transmission electron microscope (TEM), the DNA content and mitochondrial transmembrane potential (DeltaPsim) changes were studied by flow cytometry (FCM). RESULTS: The typical morphological changes of LEC apoptosis in Cur group detected by TEM included chromatin condensation and aggregation at the periphery of the nucleons and nuclear fragmentation. The DNA content of LEC in Cur group decreased time-dependently. The DNA content was significantly lower than that of the control group (P < 0.01). The DeltaPsim of LEC in Cur group was decreased, appeared in early stage (8 hours) and reached the maximum after 72 hours. The difference of DeltaPsim of LEC between Cur group and the control group was significant (P < 0.01). CONCLUSIONS: Cur can remarkably induce apoptosis of LEC in vitro. Cur induced LEC apoptosis is caused by decrease of DNA content in LEC nucleus. Collapse of DeltaPsim in cytoplasm induced by Cur results in the irreversible apoptosis process of LEC. This is the early event of LEC apoptosis. LEC apoptosis induced by Cur may pass through two pathways: nuclear pathway and cytoplasmic pathway. The apoptosis of LEC induced by Cur may be the cellular and molecular mechanisms of reducing lens posterior capsular opacification by Cur. Cur may become an effective and low toxic medication for the prevention and treatment of after-cataract. PMID- 17081434 TI - [The genetic engineering animal models of retinoblastoma]. AB - The article summarized the latest progress of retinoblastoma animal models using transgenetic engineering technology, and discussed the mechanism of Rb production. According to the the established animal models using current genetic engineering, the dysfunction of Rb1 gene is not the only gene in response to the development of Rb, multiple genes abnormal are required. PMID- 17081435 TI - [Emphasis on basic research for corneal diseases]. AB - Corneal disease is the second leading cause of blindness in China. Improvement of the diagnosis and treatment of corneal disease depends on the deepening and widening of the basic research in this area. In some fields, the research conducted in our country has reached the top level in the world. However, in general, obvious disparities exist between China and the developed countries in terms of lack of systematic research, limited financial support, the insufficiency of the quality and quantity of researchers and so on. Based upon the current situations of China, the future basic research on corneal diseases should focus on the mechanism of infectious corneal diseases, gene mutation of corneal dystrophy, corneal tissue engineering, the pathogenesis of dry eye, the reconstruction after severe chemical burns of ocular surface, the pathogenesis of ocular surface neovascularization, the prophylaxis and therapy of corneal graft rejection, etc. Other important measures to improve the basic research of corneal diseases in our country would include applying for more research funds, training high quality researchers, and strengthening the cooperation with different disciplines and with other countries. PMID- 17081436 TI - [Study of limbal stem cells and related problems]. AB - Basic and clinical research of limbal stem cells has enhanced the understanding of ocular surface diseases. This article analyzes the value and problems existing in the identification of limbal stem cells by anatomical location, proliferation capacity and the use of differentiating surface markers. The clinical methods for the diagnosis of limbal stem cell deficiency and the effects of limbal epithelial cell transplantation are reviewed. Some key problems that must be solved in the future such as the identification marker of limbal stem cells and the final outcome of transplanted cells are presented. PMID- 17081437 TI - [Transplantation of bioengineered corneal epithelium for the treatment of total limbal stem cell deficiency in rabbit]. AB - OBJECTIVE: To study the results of transplantation of cultured autologous limbal stem cells using fibrin gel membrane as substrate for the treatment of limbal stem cell deficiency in rabbits. METHODS: An ocular surface defect was created in the right eyes of all rabbits by a lamellar keratectomy extending 1 mm outside the limbus. The stem cells were isolated from a small limbal biopsy specimen from the left eyes and cultured. Cells were grown over fibrin gel membrane to construct bioengineered corneal epithelium. After cytologic verification of total limbal stem cell deficiency, rabbits were divided into 4 groups at random. The fibrovascular pannus of each cornea was removed. Groups I - III (n = 24) underwent transplantation of limbus stem cells cultured on fibrin gel membrane and were observed for 3 months (Group I), 1 month (Group II) and 2 weeks (Group III), whereas group IV (n = 8) received only the fibrin gel membrane. Clinical outcome was graded by corneal integrity, opacity and neovascularization. HE staining, impression cytology and immunofluorescence staining with antibodies against keratin-3, MUC5AC and nuclear p63 were performed to assess the phenotype of corneal epithelium. RESULTS: The cornea in the study groups (group I - III) recovered smoothly, covered with transparent epithelium and without neovascularization. In the control group (group IV), cornea was turbid and irregular, with neovascularization (P = 0.021). Corneal impression cytology indicated PAS (-) in the study groups, while PAS (+) in the control group. HE staining of the cornea in the study groups showed normal corneal phenotypes; whereas the epithelium cells in the control group showed conjunctival phenotypes with vessels and goblet cells. Immunostaining of study groups showed keratin-3 positive and MUC5AC negative corneal epithelium, and p63 was expressed in the basal cell of corneal epithelium. Cells in the control group expressed MUC5AC abundantly. CONCLUSIONS: Tissue bioengineered cornea epithelium transplantation can successfully reconstruct corneal surface affected by limbal stem cell deficiency. Fibrin gel membrane is a new bioengineered material with the characteristics of absorbability, transparence and good histological compatibility. It is supposed to be an optimum stem cell scaffold. PMID- 17081438 TI - [The transfection and expression of IL-1ra gene to the rabbit cornea in situ via cation polymer mediation]. AB - OBJECTIVE: To investigate the efficiency and safety of transfection of PEGFP-IL 1ra plasmid via cation polymer mediation (poly-ethylenimine, PEI) by injection into the corneal stroma. METHODS: Human IL-1ra cDNA fragments were cloned by RT PCR. Plasmid PEGFP-hIL-1ra recombinants were constructed and transferred into corneal endothelial cells (CEC) via cation polymer mediation. Expression of IL 1ra mRNA and IL-1ra was detected by green fluorescent protein (GFP) and Western blotting. In the experiment group, 20 microl preparation containing 10 microg plasmid PEGFP-hIL-1ra recombinants and PEI-in-vivo was injected into the corneal stroma of Wistar rats (n = 30). Equivalent PEI-in-vivo solution was injected into another 15 corneas as the controls. Corneas were harvested at different time points (day 1, 3, 6, 14 and 21) after injection. The changes of tissue structure and function after IL-1ra in situ transfection were studied by HE staining, transmission electron microscopy, trypan blue-alizarin red staining and immunohistochemistry. The location and intensity of IL-1ra-GFP fusion protein expression were monitored by fluorescence microscopy. RESULTS: The size of the RT PCR product of hIL-1ra fragments was approximately 500 bp in agarose gel electrophoresis. Restrictive enzyme digestion analysis of PstI, BamHI and DNA sequence analysis showed that expression of plasmid PEGFP-hIL-1ra recombinants had been constructed successfully. Twelve hours after the transfection of PEGFP hIL-1ra, GFP fluorescence was detected in 10% - 15% endothelial cells. IL-1ra protein (RMW: 44,000) was detected by Western-blotting. In PEGFP-hIL-1ra treated group, fluorescence was appeared at day 1 in cornea basal epithelial cells, peaked at day 6 in whole cornea, began to weaken at day 14, and only weak fluorescence remained in cornea epithelial cells at day 21. No fluorescence appeared in the control group. No significant pathologic changes could be found in HE stained cornea tissues in both transfected group and the controls. p63 immunocytochemical staining in cornea epithelium was positive in both groups. Trypan blue-alizarin red staining confirmed that there was no damage in cornea endothelial cells. IL-1ra-GFP granules could be found by transmission electron microscope in every layer of cornea in the transfected group, but none in the controls. There was no impairment in the ultrastructure of cells in both groups. CONCLUSIONS: By direct injection of PEGFP-hIL-1ra into corneal stroma and mediated by cation polymer, IL-1ra genes could be transferred and expressed in corneal tissue efficiently and safely, and might provide a novel technique of gene transfection to cornea in situ. PMID- 17081439 TI - [Histopathological study of corneal wound healing in guinea pigs after deep lamellar endothelial keratoplasty with small incision]. AB - OBJECTIVE: To study the histopathological characteristics of corneal wound healing after deep lamellar endothelial keratoplasty (DLEK) with small incision. METHODS: Forty-five guinea pigs were divided into the donor and recipient groups, which consisted of 15 and 30 guinea pigs respectively. DLEK was performed in the right eye, and the left eye was used as histopathological control. Two guinea pigs with clear right corneas under biomicroscopy examination were killed at 1 week, 2 weeks, 1 month, 2 months, 3 months and 4 months postoperatively, separately. Bilateral corneal buttons were harvested for the evaluation of endothelial cells (alizarin red stain) and for the light microscopy study (HE or PAS staining). RESULTS: Clear cornea could be obtained in 22 recipient eyes postoperatively. In these corneas, the endothelial cells either in the donor disc or recipient's bed were normal in shape and size. However, there was a band area between the donor disc and recipient's bed, where the endothelial cells were abnormal. In the early postoperative period, histopathological study disclosed increase of number of cells in the corneal stroma along the pathway of lamellar separation. The tissue connecting the gap between the donor and recipient lacked lamellar structure. The new descemet's membrane appeared beneath the endothelium 1 month postoperatively. Three to four months after the operation, the continuity of the descemet's membrane and the posterior lamella was rehabilitated. CONCLUSIONS: The corneal wound healing process after the DLEK with small incision is related with the type of trauma. Collagen is arranged well at the surface of the lamellar. Fibrotic repair is seen in the marginal area connecting the donor disc and recipient's bed. PMID- 17081440 TI - [The expression of nuclear factor-kappa B in lipopolysaccharide-induced keratitis of rats and the effect of pyrrolidine dithiocarbamate on its expression]. AB - OBJECTIVE: To investigate the expression of nuclear factor-kappa B (NF-kappaB) and cytokines in lipopolysaccharide (LPS)-induced keratitis of rats, and the effect of pyrrolidine dithiocarbamate (PDTC) on its expression. METHODS: A LPS induced keratitis model was established in Wistar rats. Thirty minutes before LPS exposure, PDTC and normal saline were injected into subconjunctival tissues separately. At 0.5, 1.0, 3.0, 6.0, 12.0, 24.0 and 72.0 h after LPS exposure, the rats were examined with slit-lamp microscope. Then they were sacrificed and the corneas were excised for routine histological analysis. Immunohistochemical staining with an antibody against activated NF-kappaB was performed to detect the expression of NF-kappaB. The change of TNF-alpha mRNA expression was identified by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: Histology findings demonstrated that corneas exposed to LPS showed significant changes in corneal structure, edema and pronounced inflammatory cells infiltration were observed. Both symptoms and damages of the cornea were lesser in the rats of PDTC group than that of the control keratitis group. Compared with PDTC group, the activation of NF-kappaB and the expression of TNF-alpha mRNA were significantly upregulated after LPS challenge 0.5 - 24.0 h (P < 0.01), separately; peak expression of NF-kappaB and TNF-alpha mRNA were observed at 3.0 - 12.0 h after LPS exposure. CONCLUSIONS: The expression of NF-kappaB and TNF-alpha mRNA induced by NF-kappaB plays an important role in the pathogenesis of keratitis. The inhibitor of NF-kappaB, PDTC, can relieve the cornea from damages produced by keratitis. PMID- 17081441 TI - [Analysis of causes for penetrating keratoplasty at Shandong Eye Institute from 1997 to 2002]. AB - OBJECTIVE: To examine the major causes and identify trends of penetrating keratoplasties (PKPs) performed in Shandong Eye Institute in the past 6 years. METHODS: The patients who received PKP in Shandong Eye Institute between January 1, 1997 and December 31, 2002 were reviewed retrospectively. The causes were classified into 10 different groups: purulent keratitis, herpes simplex keratitis (HSK), keratoconus, bullous keratopathy, corneal scar, corneal trauma, corneal burn, regraft, corneal dystrophy and degeneration, and others. The trend of changes in causes was analyzed. Purulent keratitis was further classified according to the pathogens. The initial diagnosis and causes of regrafting were noted, as well as the intraocular surgeries related to bullous keratopathy. RESULTS: A total of 1702 cases were included in this study. The leading indications for PKP, in order of decreasing frequency, were purulent keratitis (31.9%), HSK (17.6%), keratoconus (12.9%), corneal scar (9.6%), bullous keratopathy (6.9%), penetrating corneal trauma (6.2%), regraft (4.5%), thermal and chemical burn (4.1%), corneal dystrophy and degeneration (3.9%) and others (2.4%). An increasing trend in keratoconus (chi(2) = 26.399, P < 0.001) and bullous keratopathy (chi(2) = 11.4229, P < 0.001) was noted, but not in the others. Fungi were the most common pathogens of purulent keratitis (65.9%). No changes were found in the pathogens of purulent keratitis during the past 6 years. Most of bullous keratopathy had a history of cataract operation (76.3%). Ocular burn (24.7%), HSK (23.4%) and purulent keratitis (14.3%) were the three primary affections of regraft. The main causes of graft failure that lead to the regraft were immune rejection (61.0%), grafts infection (14.3%) and HSK recurrence (10.4%). CONCLUSIONS: Infectious keratopathy is still the leading indication for PKP. Among the infectious keratitis, fungal infection is the top cause of PKP. Keratoconus and bullous keratopathy have shown an increasing trend. PMID- 17081442 TI - [Anterior chamber phakic intraocular lens implantation in highly myopic eyes]. AB - OBJECTIVE: To investigate the stability, predictability and safety of angle supported phakic intraocular lens (IOL) implantation in high myopia. METHODS: Seventy-six myopic eyes of 40 patients treated with anterior chamber angle supported phakic IOL (Phakic6H) implantation were followed. The average myopia diopter was (-15.89 +/- 3.78) D (-9.50 to -26.25 D) and the best corrected visual acuity (BCVA) was (0.82 +/- 0.20) (0.5 to 1.0). The average follow-up time was 12 months. All the treated eyes were observed for anterior segment changes, uncorrected visual acuity (UCVA), BCVA, refraction, intraocular pressure (IOP) and corneal endothelium preoperatively and postoperatively. All data were analyzed in statistics. RESULTS: One year after the surgery, the average UCVA was (1.00 +/- 0.35) (0.3 to 1.5) and BCVA was (1.18 +/- 0.29) (0.5 to 1.5). The average residual refraction was (-0.40 +/- 0.64) D (-2.00 to +0.50 D). In 96.1% of eyes, the average residual refraction was within +/- 1.0 diopter. The average corneal endothelial count before and one year after the surgery was (3174 +/- 248)/mm(2) and (3067 +/- 320)/mm(2) (P < 0.01) respectively. The intraocular pressure before and one year after the surgery was (16.12 +/- 2.32) mmHg and (15.29 +/- 3.38) mmHg (P < 0.05) respectively. Two patients (4 eyes) complained of halo and glare. One patient's (2 eyes) pupils were oval and another one (2 eyes) showed decentered phakic IOL. Macular hemorrhage occurred in one patient (2 eyes) after eight months of surgery. CONCLUSIONS: Angle-supported phakic IOL implantation is effective for treatment of high myopia and the results are predictable. It deserves further clinical investigation. The effectiveness and safety of this kind of phakic IOL should be further observed for a long period of time. PMID- 17081443 TI - [Clinical analysis of measuring corneal thickness with non-contact method]. AB - OBJECTIVE: To compare accuracy of the Pentacam with A-ultrasonoscope in the measurement of central corneal thickness. METHODS: According to the degree of myopia, 107 myopic patients (211 eyes) were divided into four groups. The central corneal thickness was measured with the Pentacam system and A-ultrasonoscope. The results were compared statistically and the differences between these two methods were analyzed. RESULTS: Mean values of central corneal thickness measured by the Pentacam system and A-ultrasonoscope was (535.58 +/- 36.34) microm and (536.73 +/ 37.18) microm, respectively. There was no difference in the results obtained by these two methods in each myopia group (P > 0.05). The 95% limits of agreement were (-1.88, -0.42). CONCLUSIONS: The data shows that the results obtained by Pentacam system do not have significant difference as compared with that from the A-ultrasonoscope. The Pentacam system can provide precise results and has the advantage of simplicity and non-contact. Therefore, it would be a promising instrument for the evaluation of central corneal thickness before the cornea refractive surgery. PMID- 17081444 TI - [Diagnosis and differential diagnosis of acute zonal occult outer retinopathy]. AB - OBJECTIVE: To study the clinical manifestations of acute zonal occult outer retinopathy (AZOOR) and to differentiate it from other retinal diseases. METHODS: Six patients diagnosed AZOOR had complete eye examinations including fundus photography, fundus fluorescein angiography (FFA), electroretinography (ERG), visual evoked potentials and visual field examination. Medical consultation and neurological consultation were performed in those patients. All patients were followed up and the data were collected for analysis, discussion, diagnosis and differential diagnosis. RESULTS: Six patients (five female and one male) aged 26 42 years (mean 35 years) with AZOOR were followed up for 4 - 18 months [mean (7.5 +/- 3.2) months]. All of them were affected bilaterally and their visual acuity were slightly reduced except one eye was CF/40 cm. Half of them had photopsia. At least one eye of each patient had visual field defect or decreased sensitivity in local area or blind-spot enlargement. Biomicroscopic examination revealed vitreous cells in 10/12 eyes and anterior chamber inflammatory cells and keratic precipitate in 4/12 eyes. Minimal (10/12 eyes) or no (2/12 eyes) fundus changes were found in their initial examination. Funduscopic examination revealed yellow-white dots (4/12 eyes) and gray dots (6/12 eyes) at the posterior pole of deep retina or retinal pigment epithelium-Bruch membrane-choroid capillary complex layer. FFA showed depigmentation (2/12 eyes) or hyperfluorescein spots (10/12 eyes) that identical to the retinal lesions. In the follow-up examination, the visual acuity was reduced in one eye and visual field defect enlarged in both eyes of one patient; the number of retinal dots increased in one eye, decreased in one eye and extinguished in one eye. ERG or mERG revealed abnormal in all of their eyes with no changes in their follow-up examination. All of the initial diagnoses of six patients were not consistent with final diagnosis. CONCLUSIONS: AZOOR is a rare eye disease, usually occurs in young females, with the characteristics of photopsia, visual field defects, abnormal ERG and slight changes in the fundus. The differential diagnosis of this disease is relatively complicate and is easily to be misdiagnosed. PMID- 17081445 TI - [A comparative study of various diagnostic procedures in macular edema]. AB - OBJECTIVE: To compare the results for diagnosis of macular edema between Heidelberg retina tomography (HRT-II macular edema module) and optical coherence tomography (OCT). METHODS: Two hundred and twelve eyes of 106 macular edema patients were diagnosed with fundus fluorescein angiography (FFA). HRT-II was used to obtain Edema Index and OCT was used to measure the retina thickness in the fovea. The results were compared with FFA. RESULTS: As FFA for gold standard, we compared with other diagnosis method. With regard to the diagnosis of macular edema by OCT, the Kappa value was 0.553, the odds ratio (OR) was 8.519. The Kappa value of HRT-II was 0.403 and OR was 3.210. In two groups of patients with and without macular edema, which were diagnosed by FFA, the difference of Macular edema Index P assayed by HRT-II between these two groups was significant (P = 0.00); while the difference of fovea retina thickness P assayed by OCT between these two groups was significant, too (P = 0.00). The best critical value of E was 2.00 and the ROC curve's area was 0.673 in the detection of macular edema by HRT-II. The best critical value of retina thickness for the diagnosis of macular edema was 170 microm and the ROC curve's area was 0.774 in the detection of macular edema by OCT. CONCLUSIONS: In regard to the diagnosis of macular edema, the sensitivity of HRT-II is better than that from OCT, with a lower false negative result. However, HRT-II has a relatively low specialty with a higher false- positive result. As for the localization of macular edema, HRT-II has its advantage. Compared with FFA, the consistence of OCT for the diagnosis of macular edema is better than that of HRT-II. We can conclude from the OR value that OCT has greater influence than HRT-II in the diagnosis of macular edema. In addition, as for the diagnosis and different diagnosis, the clinical value of OCT is higher than that of HRT-II. Combination of these two different diagnostic methods might provide an accurate quantitative analysis of macular edema. PMID- 17081446 TI - [Detection of mtDNA*LHON G11778A mutation by real-time polymerase chain reaction using TaqMan-MGB probe technology]. AB - OBJECTIVE: To develop a simple, rapid and reliable real-time PCR assay based on TaqMan technology using a new MGB probe for detecting mtDNA(*)LHON G11778A mutation and heteroplasmy directly. METHODS: Twenty patients with suspicion of Leber hereditary optic neuropathy (LHON) and their maternal relatives had undergone molecular genetic evaluation. Seventeen normal individuals were used as the controls. A real-time PCR involved two MGB probes (wild-type and mutation type) in a single tube on the iCycler IQ real-time detection system was used to detect the mtDNA(*)LHON G11778A mutation. The results were then compared with the DNA sequence analysis of the PCR products. A linear standard curve was obtained by pUCm LHON-G and pUCm LHON-A clone. RESULTS: In the controls (wild type), the reaction of VIC-labeled MGB probe was positive and the channel of FAM reaction was negative, the DNA sequence was 100% matched to previously published data. In 20 LHON patients and their maternal relatives, 12 cases showed mutations in DNA sequence analysis, all of them were LHON mtDNA mutation. While 5 other cases showed the combination of LHON mtDNA mutation and wide type gene phenotype, the rate of Ct value in wild type versus gene mutation was over 25%. DNA sequence analysis showed 8 of LHON mtDNA belonged to wild types and 3 cases were heteroplasmy, and the rate of Ct value in gene mutation versus wild type was lower than 25%. CONCLUSION: This real-time PCR assay is a simple, rapid and reliable method for the detection of genotyping mtDNA mutations as well as for quantifying heteroplasmy. PMID- 17081447 TI - [A preliminary study in establishment of mice model of experimental uveal melanoma]. AB - OBJECTIVE: To explore the methods to establish a nude mice model of experimental uveal melanoma by the implantation the primary cultured cells into anterior chamber or subcutaneous injection. METHODS: 35 nude mice, which applied with the SPF grade standard by national healthy department, were divided into 3 groups: anterior chamber group (15), subcutaneous A group (10) and subcutaneous B group (10), respectively. (2 - 3) x 10(5) melanoma cells were inoculated into the anterior chamber of each nude mouse, while 2 x 10(6) cells were inoculated into every subcutaneous of the hind legs in A group. Besides this, the fresh mass of the tumor come from enucleated eye were transplanted into every subcutaneous of the hind legs in B group. Then the condition of transplanted tumor were observed under the slit lamp and naked eye, all nude mice were followed for 3 months the rates of tumor induction were compared. RESULTS: The results of the rates of tumor induction in the 3 groups were 7 (46.6%) of 15, 2 (20%) of 10 and 0 of 10, respectively. The discrepancy were of significance (chi(2) = 7.080, P = 0.029). CONCLUSIONS: Uveal melanoma can be induced successfully by inoculation the primary cultured human uveal melanoma cells into anterior chamber or subcutaneous in nude mice. The results suggest that the ration of the tumor growth is higher in subcutaneous mass transplant than any other groups. PMID- 17081448 TI - [Changes of protein kinase C for the proliferation of retinal pigment epithelium cells induced by subretinal fluid]. AB - OBJECTIVE: To study the effects of subretinal fluid (SRF) on the proliferation of retinal pigment epithelium (RPE) cells and on the activation and translocation of protein kinase C (PKC); to investigate the relationship between the proliferation and the changes of PKC in the RPE cells; and to study the effect of PKC inhibitor. METHODS: RPE cells were harvested to measure the PKC activity in cytoplasm and cellular membrane after being treated with subretinal fluid (SRF) obtained from PVR patients with different degrees (B grade and C), PKC specific activator PMA (positive control) or normal vitreous. RPE cells cultured with DMEM culture medium only were used as the negative controls. PKC activity in cytoplasm and cellular membrane was measured with radioactive isotope (32)P label method. For further study, the PKC specific inhibitor N, N-dimethyl was used to pretreat the RPE cells before the administration of SRF, PMA or normal vitreous, and then the activity of PKC was observed and recorded. (3)H-TdR was used to measure the proliferation of RPE cells with or without the activation and translocation. RESULTS: SRF and PMA could promote the proliferation of RPE cells and activate PKC in the cytoplasm of RPE cells, and then promoted the PKC translocated from the cytoplasm to cellular membrane. The peak of PRC on the cell membrane appeared later in cells treated with SRF when compared with those treated with PMA. The appearance of peak of PKC in cells treated with SRF from grade B PVR was later than those treated with SRF from grade C PVR, the stimulating effect on the proliferation of RPE cells by the SRF B was also less than those from the SRF C. No activation of PKC and increased proliferation were observed in RPE cells treated with normal vitreous or DMEM culture medium. Pre-treatment with PKC inhibitor could block the PKC-activating effects and proliferation-stimulating effects of SRF and PMA on the RPE cells, therefore no significant difference could be detected between different groups. CONCLUSION: SRF can promote the proliferation of RPE cells and induce the activation and translocation of PKC in RPE cells. PKC is involved in the process of the proliferation of RPE cells. The PKC inhibitor could block this process. PMID- 17081449 TI - [Cloning, expression and cytotoxin to human lens epithelial cells of targeting toxin DT389-hbFGF]. AB - OBJECTIVE: To construct pGEX-DT(389)-hbFGF plasmid, express and identify the cytotoxicity to human lens epithelial cells (HLECs). METHODS: Extracting DNA of dead diphtheria bacillus and RNA of 12-week fetal brain cortex. The fragments of truncated diphtheria toxin (containing 389 amino acids of N-terminus, DT(389)) and full length human bFGF gene (encoding 18kd protein) were amplified by PCR technique respectively. The two fragments were inserted into prokaryotic expression vector pGEX-4T-1. After testing sequence, the expressing plasmid was transformed into E.Coli BL21 strain and induced expression under IPTG. The expressed fusion protein was purified and identified. MTT experiment tested cytotoxicity of the fusion protein to HLECs in vitro. The way of HLECs death under different dosage was identified by flow cytometry. RESULTS: The gene fragments of DT(389) and human bFGF were accurately amplified. The expression vector including DT(389)-hbFGF fused gene was constructed and expressed successfully. DT(389)-hbFGF fusion protein can induce HLECs apoptosis in a dosage dependence manner during certain range. The LD(50) was about 3.8 x 10(-11) mol/L. CONCLUSION: The successful cloning and expression of DT(389)-hbFGF immunotoxin lay a foundation for accelerating lens epithelial cells apoptosis and the targeting therapy toward posterior capsule opacification. PMID- 17081457 TI - [Non-fusion technique on the surgical treatment of spinal degenerative disease]. PMID- 17081459 TI - [The future of spine functional reconstruction--from fusion to non-fusion]. PMID- 17081460 TI - [Anterior cervical corpectomy and fusion with preserved posterior vertebral wall for multi-level cervical spondylosis: a prospective random study of 84 cases]. AB - OBJECTIVE: To compare the results of anterior cervical corpectomy and fusion with preserved posterior vertebral wall (PWCF) with the classical anterior cervical corpectomy with fusion (ACF). METHODS: Eighty-four patients with multi-level cervical spondylosis were randomized to ACF (n = 42) or PWCF (n = 42). Postoperative JOA score were evaluated by an independent observer. Fusion rate, segmental lordosis, and disc height were assessed by radiographs at postoperative 3 months, 6 months, and 12 months. Fusion evidence was observed by CT scan. RESULTS: JOA score improvement was similar for both treatments. The operation time was shorter in the PWCF group than in the ACF group. The intraoperative blood loss was less in the PWCF group than in the ACF group. Fusion rate was 100% for both groups 3 months after operation. The segmental lordosis and the disc height increased were similar in both groups. CONCLUSIONS: Except for reduced risk, time and blood loss during the operation, PWCF is the same as for ACF procedure. Reservation of posterior vertebral wall results in more reliable fusion. But replication of the results by other team and intensive assessment still need to be carried out. PMID- 17081461 TI - [Analysis of the long-term outcome of anterior approach surgery on cervical spondylotic myelopathy]. AB - OBJECTIVE: To investigate the long-term efficacy of anterior approach surgery on cervical spondylotic myelopathy and factors affecting prognosis. METHODS: The data in 116 patients suffered from cervical spondylosis from January 1992 to December 2000 were reviewed, including 80 male cases and 36 female cases, whose age ranged from 36 to 76 years (mean, 51 years). The preoperative course of disease was 2 months to 20 years (mean, 19 months). There were 65 cases (56.0%) with single segments involved, 44 cases (37.9%) with two segments, 7 cases (6.0%) with three segments. Ninety-eight cases were onset slowly, 18 cases with no remote cause and aggravating quickly. Three kinds of surgeries were performed: anterior cervical decompression and autoiliac bone interbody fusion, anterior cervical decompression and fusion with threaded fusion cage, anterior cervical decompression and autoiliac bone interbody fusion with anterior screw-plate system. Improvement in spinal cord function was assessed using the Japanese Orthopaedic Association (JOA) scoring system, the long-term efficacy and influential factors were also analyzed. RESULTS: The mean follow-up time was 7 years and three months (5 - 12 years). The mean preoperative JOA score was 9.34 +/- 1.81. The mean postoperative JOA score was 10.35 +/- 1.85. At the final follow-up, the JOA score was 14.09 +/- 1.90 and the recovery rate was 63.2%. Among the total patients, 27 cases were excellent, 47 cases were fine, 23 cases were good, 19 cases were poor, the fineness rate was 63.8%. The long-term efficacy of anterior approach surgery has close correlations with time of course, age of onset, preoperative spinal cord function and the number of affected segments, but has no correlations with modes of fusion and internal fixation. CONCLUSIONS: The patients will be attentively observed while having a definite diagnosis of cervical spondylotic myelopathy. The good long-term results will be obtained after early anterior cervical decompression and fusion. PMID- 17081462 TI - [A comparative study on maintenance of disc height by different anterior cervical fusion methods]. AB - OBJECTIVE: To compare the effect of autograft, cage and autograft plus internal fixation with plate on maintenance of disc height and recovery of spinal cord function. METHODS: From March 1998 to June 2004, 117 cervical spondylotic patients who received anterior fusion surgery were reviewed. There were 65 males and 52 females with an average age of 57 years (range from 31 to 72 years). One level was involved in 91 patients, and two levels in 26. Fusion methods included autogenous tri-cortical iliac bone grafting alone in 31 patients, cage with autograft bone in 38 and autogenous tri-cortical iliac bone grafting combined with self-locking plate in 49. The measured fusion length was determined as the distance from the midpoint between the anterior and posterior aspects of the superior endplate of the top level of the fusion to the midpoint of the inferior endplate of the bottom level of the bottom level of the fusion. JOA rating criteria was used for cord function evaluation. One-way variation analysis was used to compare the disc height loss and cord function among the 3 groups. RESULTS: All patients were followed-up for an average period of 23 months (range, 13 to 59 months). At final follow- up, disc height loss averaged 1.94 mm in autogenous tri-cortical iliac bone grafting group, greater than that in the other two groups (1.48 mm in cage group and 1.25 mm in instrumented group) with statistical significance. Recovery of spinal cord function was satisfactory in all groups at one year postoperatively with no statistical difference. Three implants failure occurred in self-locking plate group and was salvaged with uneventful recovery. CONCLUSIONS: In anterior cervical fusion surgery, maintenance of disc height could be achieved by proper application of cage or internal fixation with plate. PMID- 17081463 TI - [A method to avoid the fixator failure by using pedicle screw combined vertebroplasty for spine fractures]. AB - OBJECTIVE: To study a new implant material (carbonated hydroxyapatite, CHA) united pedicle screw to cure spine fracture. METHODS: Thirty-two cases of spine compressed fracture were used with pedicle screw fixator and vertebroplasty. Before operation, patients' vertebral body were compressed (46 + 21)% (20% approximately 70%) on average. In operation, broken vertebral body was reposition through pedicle screw technique, then used self-made syringe to inject CHA into anterior and central column of broken vertebral body through pedicle. And all of patients were not given any bone-graft. RESULTS: In 6 - 26 months followed-up, no immunologic rejection was found about hydroxyapatite, and no any broken of the screws and shafts was found, no loosing and other complications either. All the patients could move in 3 - 5 days after operation. The height of the broken vertebral body were reduced 97% compared with pre-operation. And CHA in vertebral body was degraded gradually, and at the same time it was replace by new bone in vertebral body. After operation, VAS score was 61 +/- 32, and there was significant difference compared with pre-operation. CONCLUSIONS: The pedicle screw fixation united vertebroplasty is an efficient way to prevent the failure of the treatment of spine fracture. PMID- 17081464 TI - [Influence of preoperative range of motion on the early clinical outcome of total knee arthroplasty]. AB - OBJECTIVE: To retrospectively analyze the influence of preoperative range of motion (ROM) and maximal flexion degree on the early clinical outcome of total knee arthroplasty (TKA). METHODS: From January 2000 to December 2003, 97 knees of 65 patients that were underwent total knee arthroplasty with Scorpio posterior stabilized knee prosthesis were reviewed. There were 55 osteoarthritis patients (81 knees), and 10 rheumatoid arthritis (16 knees). Thirty-three patients were underwent unilateral TKA, 32 patients were underwent bilateral TKA. According to the preoperative ROM of knee, these patients were divided into two groups, one 90 degrees (range, 95 degrees - 140 degrees ). Finally the clinical outcomes of two groups (include ROM, maximal flexion degree, KSS score and function score) were evaluated. Three days later after operation, continuous passive motion (CPM) and active functional exercise of the knee were begun, and the wound healed well in all patients. All these operations were primary total knee arthroplasty. RESULTS: The patients were followed up for average 2 years 5 months (range, 10 months to 3 years 8 months). The average ROM of knee was improved to 101.6 degrees (range, 40 degrees - 140 degrees ) after operation from 84.2 degrees (range, 5 degrees - 140 degrees ) before operation (P = 0.000); the average maximal flexion degree was decreased from 103.5 degrees (range, 25 degrees - 140 degrees ) before operation to 101.6 degrees (range, 40 degrees - 140 degrees ) after operation (P = 0.439); KSS of knee joint was improved to 78.8 points after operation (range, 50 - 95 points) from 19.5 points (-24 - 62 points) before operation (P = 0.000). There was statistically difference between the clinical outcomes (ROM, maximal flexion degree, KSS score and function score) in the two groups before and after operation. Those knees with good preoperative ROM tend to lose flexion, while those with poor preoperative ROM gain flexion after TKA. No revision and deep infection happened. CONCLUSIONS: TKA is a complex operation, the clinical outcome of TKA is mainly determined by the good operation skill, abundant clinical experience and the familiarity with the prosthesis of the surgeon. At the same conditions such as same surgeon, same prosthesis and same physical therapy, preoperative range of motion of knee influence on the early clinical outcome of total knee arthroplasty, knees that have good preoperative ROM have better clinical outcomes postoperatively than those with poor preoperative ROM. PMID- 17081465 TI - [Establishment of arthroscopic trans-septal approach and its clinical application]. AB - OBJECTIVE: To investigate the method and result of arthroscopic trans-septal approach (ATS). METHODS: Ten fresh cadaveric knees were prepared for anatomical study about the posterior septum, and 65 posterior compartment arthroscopy of the knees were performed to view the structure of the posterior septum. The initial diagnosis included: rheumatoid arthritis, pigmented villonodular synovitis, osteoarthritis, loose body or foreign body in the posterior compartment, posterior cruciate ligament (PCL) injury or avulsion fracture, posterior horn tear of meniscus, undiagnosed swollen knee with pain and effusion, osteochondritis dissecans, pyogenic arthritis, gout. From January 2002 to June 2005, 22 cases of ATS were applied. Anterolateral portal was initially created, followed by posterolateral portal under the viewing of arthroscopy which was located at the anterolateral portal. Anteromedial and posteromedial portals were also created using the same technique. Arthroscopy was then transferred to the posteromedial portal, and blade was introduced from the anteromedial portal to gradually remove the synovium covering PCL. Arthroscopy was relocated to the anteromedial portal, Wissinger rod was introduced from the posteromedial portal and pointed to the posterior septum adjacent to the posterior edge of the midportion of PCL. The Wissinger rod was pushed carefully to pierce through the posterior septum under the sight of arthroscopy which was located at the posterolateral portal. ATS was finally created. RESULTS: The posterior septum was in the middle of posterior compartment of the knee, which was film screen-like at the sagittal plane and sandwich-like at the transverse plane. The synovium covered the posterior septum at arthroscopic inspection. Twenty-two cases of ATS were successfully created, amounting to 34% (22/65) of all cases at the same period which had received the arthroscopy of posterior compartments of the knees. Synovectomy of the posterior compartments of the knees was performed in 7 cases, loose body removal was in 6 cases, PCL reconstruction was in 4 cases, reduction and fixation of PCL avulsion fracture was in 2 cases. Chondroplasty, inflammatory synovectomy, and meniscectomy were performed accordingly in 6 osteoarthritis cases. No vascular or nervous injury was encountered. At an average of 20 months follow-up (range, 4 to 45 months), 9 cases still had mild knee pain or swelling, 2 cases had severe pain and were recommended for total knee replacement, the other 11 cases had no recurrence of knee pain or swelling. CONCLUSIONS: ATS has no blind area under arthroscopic vision and facilitate trans-septal operation. It is a safe and effective method to treat the diseases of the posterior compartment of the knee. The direction of inside to outside to create ATS is comparatively reliable, and PCL could be identified as an interior landmark during the passage of Wissinger rod through posterior septum to create ATS. PMID- 17081466 TI - [The use of a closed-suction drain in total knee arthroplasty: a prospective, randomized study]. AB - OBJECTIVE: To determine the value of using a closed-suction drain in cemented knee arthroplasty. METHODS: One hundred patients were prospectively randomized into two groups, one group underwent cemented total knee replacement with a single deep closed-suction drain, and the other group (n = 50) no drain. RESULTS: The total blood loss was significantly greater in those with a drain although those without lost more blood into the dressings. There was no statistical difference in the postoperative swelling or pain score, ecchymosis, deep vein thromboses (DVT), time at which flexion was regained or the incidence of infection at a minimum of two years after surgery in the two groups. CONCLUSION: There is no evidence to support the use of a closed-suction drain in cemented knee arthroplasty. PMID- 17081467 TI - [The application of three dimensional analysis system in measuring rotation angles of thumb carpometacarpal joints]. AB - OBJECTIVE: To measure the axial rotation angles of the carpometacarpal joints during the digital opposition of thumb-index finger, thumb-medial finger, thumb ring finger, thumb-little finger and the thumb's maximal opposition, then the application of these parameters were studied. METHODS: Twenty neutrality occupation volunteers (female 10, male 10) with no history of hand injuries or related diseases were involved in the study. First, all the markers' 3-D coordinates were obstained using the 3D motion analysis system (EVaRT4.1) during the digital opposition movements of thumb. Then, the axial rotation angles were calculated. RESULTS: The average rotation angles of carpometacarpal joints during all kinds of digital oppositions were 29.1 degrees +/- 9.4 degrees (male), 24.8 degrees +/- 10.2 degrees (female), while the maximal rotation angles are: 35.3 degrees (male), 28.8 degrees (female). CONCLUSIONS: The combination of video based 3-D analysis system and mathematics make it possible to measure the axial rotation angles of thumb in vivo, as a result, the rotation angles of thumb carpometacarpal joints are measured precisely for the first time. These results can provide a few parameters for treatment and rehabilitation of carpometacarpal arthrositis. PMID- 17081468 TI - [Arthroscopically assisted treatment of tibial plateau fractures by tractive reduction with external fixator]. AB - OBJECTIVE: To study the clinical effect of tractive reduction with external fixator and arthroscopically assisted treatment for tibial plateau fractures. METHODS: From February 2003 to January 2005, a total of 26 cases with tibial plateau fractures were reviewed. There were 4 cases of type I fracture, 5 type II, 4 type III, 6 type IV, 5 type V and 2 type VI based on Schatzker criteria. Before operation, X-ray examination and CT scanning were done. During operation, the dissociative fragments were reconstructed by tractive reduction with external fixator arthroscopically and fixated with screws or plates. RESULTS: All the fractures were healed within 1.5 - 4 months, with no severe complications such as poor wound healing, infection or osteofascial compartment syndrome. All patients that were followed up for 7 - 21 months (mean, 16 months) showed no traumatic osteoarthritis, inversion or eversion of the knee. According to the Rasmussen scoring system, the outcome was excellent in 11 cases, good in 13 and fair in 2, with total score of 27 +/- 2. CONCLUSIONS: As for Schatzker type I - VI tibial plateau fractures, tractive reduction with external fixator and arthroscopically assisted treatment is characterized by minimal invasion, fast fracture healing. It broadens operative interspace of articular cavity, improves accuracy of reduction, is beneficial for knee joint function to recover. PMID- 17081469 TI - [The use of minimally invasive injectable graft and internal fixation in tibial plateau fractures]. AB - OBJECTIVE: To value the use of an injectable minimally invasive calcium sulfate cement for displaced tibial plateau fractures. METHODS: Thirteen patients with lateral tibial plateau fractures treated with internal fixation and bone grafting were matched with 13 patients treated using internal fixation and an injectable calcium sulfate cement. The clinical data were retrospectively analyzed. All patients were followed up for a minimum of one year. The peri-operative complication, quality of reduction, maintenance of reduction, function assessment and development of post-traumatic osteoarthritis was compared in both groups. RESULTS: Ten patients in the internal fixation and bone graft group had excellent anatomical reductions as judged on immediate post-operative radiographs but some loss of reduction on follow-up at one year was observed in 8 of the 13 (61%) cases. Twelve patients from the MIIG group had an excellent reduction on immediate post-operative radiographs but 3 (23%)demonstrated some loss of reduction of the plateau at one year follow-up (P < 0.05). CONCLUSIONS: The use of MIIG and internal fixation is associated with more favourable clinical results than conventional treatment with internal fixation and bone grafting for lateral tibial plateau fractures. PMID- 17081471 TI - [In vivo experimental study of hat type cervical intervertebral fusion cage]. AB - OBJECTIVE: To compare the characteristics of interbody fusion achieved using hat type cervical intervertebral fusion cage (HCIFC) with those of an autologous tricortical iliac crest graft, Harms cage and Carbon cage in a goat cervical spine model. METHODS: Thirty-two goats underwent C(3, 4) discectomy and fusion in which the following were used: Group 1, autologous tricortical iliac crest bone graft (8 goats); Group 2, Harms cage filled with autologous iliac crest graft (8 goats); Group 3, Carbon cage filled with autologous iliac bone (8 goats); Group 4, HCIFC filled with autologous iliac graft (8 goats). Radiography was performed pre- and postoperatively and after 1, 2, 4, 8, and 12 weeks. At the same time points, disc space height, intervertebral angle, and lordosis angle were measured. After 12 weeks, the goats were killed and fusion sites were harvested. Biomechanical testing was performed in flexion, extension, axial rotation, and lateral bending to determine the stiffness and range of motion. All cervical fusion specimens underwent histomorphological analysis. RESULTS: One week after operation, the DSH, IVA and LA of HCIFC and Carbon cage were statistically greater than those of autologous iliac bone graft and Harms cage. Significantly higher values for disc space height, intervertebral angle and lordosis angle were shown in cage-treated goats than in those that received bone graft over a 12-week period. The stiffness of Harms cage in axial rotation and later bending were statistically greater than that of other groups. Radiographic and histomorphologic evaluation showed better fusion results in cage groups than in autologous bone group. CONCLUSIONS: HCIFC can provide a good intervertebral distractability and enough biomechanical stability for cervical fusion. PMID- 17081472 TI - [The feature of experimental endplate fracture in lumbar spine and its related factors]. AB - OBJECTIVE: To evaluate the feature of experimental endplate fracture in lumbar spine and its related factors. METHODS: Nineteen cadaveric lumbar motion segments aged 48 - 77 years were compressed by overload to fracture the endplate and dissected into isolated vertebrae to evaluate feature of their endplate failure. Before and after failure of endplate, radiographic tests were taken on every motion segment. The bone mineral density (BMD), bone mineral content (BMC) of the vertebral body and endplate were tested respectively before endplate fracture. RESULTS: Among 19 motion segments, 16 were fractured and accounted for 84.2% of all and fracture featured as stellate, step, depression and intrusion. Fracture concentrated on the center or anterior of superior endplate of the inferior vertebrae in one motion segment. Failure load of endplate was positively correlated with BMD, BMC of vertebral endplate. Within one vertebral body, the BMD and BMC of its superior endplate was markedly less than that of inferior endplate, on the other hand, the difference of BMD and BMC of endplate around one disc was opposite. CONCLUSIONS: Fractures usually concentrate on the center or anterior part of superior endplate of one vertebrae and are hard to be identified by conventional radiographic examination. Failure load of endplate is positively correlated with BMD, BMC of vertebral body and endplate. There might be certain relationship between feature of fracture and severity of disc degeneration. PMID- 17081473 TI - [The effect of rotational alignment of the femoral components on the patellofemoral biomechanics in total knee arthroplasty]. AB - OBJECTIVE: To study the effect of rotational alignment of the femoral components on the patellofemoral biomechanics in total knee arthroplasty (TKA) demonstrated on autopsy specimens, as the guide for surgeons to choose the correct reference axis for rotational alignment of the femoral components and to reduce the patellofemoral joint complications. METHODS: Select 9 frozen fresh human cadaver knees without gross deformities or instabilities and mount specimens on a patellofemoral joint testing jig connected to a Model 8501 Instron machine (Instron Corporation, Canton, MA). The study simulated the action of squatting from the standing position with the foot firmly planted. Standard TKA was performed in each specimen by the same senior surgeon using Nexgen LPS total knee system (Zimmer Corporation, Warsaw Indiana). Alter rotational alignment of the femoral components referenced to the transepicondylar axis and the Whiteside's line respectively. Measure biomechanics of the patellofemoral joints using Fuji prescale film at 30 degrees , 60 degrees , 90 degrees , 120 degrees of knee flexion respectively. The digital values were obtained by the handheld pressure measurement systems (FPD-305E, FPD-306E) and Autocad software. RESULTS: The rotational alignment of the femoral components paralleled to the transepicondylar axis had the best results of the peak value of the patellofemoral contact pressure (P < 0.05). There were no statistically significant differences in patellofemoral contact area (P > 0.05). But the patellofemoral contact area had the close correlations to the angles of the knee flexion and the specimens. CONCLUSIONS: Rotational alignment of the femoral components has a great influence on the patellofemoral contact pressure in total knee arthroplasty. It is reliable for surgeons to choose the transepicondylar axis as the reference axis to rotate femoral components. PMID- 17081479 TI - Treatment of osteoporosis in patients with chronic liver disease and in liver transplant recipients. AB - The pathogenesis of osteoporosis in chronic liver disease and post-liver transplantation is complex and heterogeneous. The development of hepatic osteodystrophy may be related to both increased bone resorption and decreased bone formation. Available medical treatments can be broadly classified into antiresorptive and bone-stimulating agents. Most published studies on the treatment of osteoporosis in patients with liver disease have used the commonly prescribed antiosteoporosis drugs approved for postmenopausal osteoporosis. These studies have included a small number of subjects and used bone mineral density (BMD) changes rather than fracture occurrence as an endpoint because of the short follow-up. Although the increases in BMD are promising, no intervention is proven to have antifracture efficacy in hepatic osteodystrophy. The natural history of bone disease following liver transplantation has not been fully investigated, although studies suggest that bone mineral loss is transient and generally reverses within a year following transplantation. The approach to treatment in liver transplant recipients should be targeted at preventing the early bone loss without interfering with the later recovery. Based on the available data, no single available agent can be considered as first-line therapy. In our opinion, the best treatment approach involves the elucidation of modifiable risk factors and the selection of agents targeted at the underlying derangements. PMID- 17081480 TI - Hepatic encephalopathy: a review of its pathophysiology and treatment. AB - Hepatic encephalopathy (HE) is a broad spectrum of neuropsychiatric manifestations usually affecting individuals with end-stage liver disease. The presence of HE is a poor prognostic sign, with 1-year mortality rates of almost 60%. There is much debate about the underlying mechanisms that result in this syndrome; however, elevated plasma and central nervous system ammonia levels are considered key factors in its pathogenesis. Initial evaluation of the patient presenting with overt HE should include a careful search for predisposing factors, including underlying infection, gastrointestinal (GI) bleeding, electrolyte disturbances, hepatocellular carcinoma, dehydration, hypotension, and excessive use of benzodiazepines, psychoactive drugs, or alcohol. The mainstay of treatment for many years has been nonabsorbable disaccharides, particularly lactulose. Alternative treatments, which usually are second line in patients who do not respond to lactulose, include zinc, antibiotics (neomycin, metronidazole, and rifaximin), ornithine aspartate, sodium benzoate, probiotics, and surgical intervention. Accepted treatments for HE are associated with significant unpleasant side effects, including diarrhea, renal failure, neuropathy, and other GI disturbance. Newer therapies are still in development, and most are awaiting human trials in order to confirm their benefit. These include manganese chelators, L-carnitine, N-methyl-d-aspartate receptor antagonists, blood purification dialysis system, and an intravenous combination of sodium benzoate and phenylacetate. PMID- 17081481 TI - Diagnosis and treatment of hepatic sarcoidosis. AB - The presence of granulomas in the liver raises consideration of a wide differential diagnosis, but in most Western series, sarcoidosis accounts for a majority of cases. This review will focus specifically on the diagnosis of and therapy for hepatic sarcoidosis. Sarcoidosis is a systemic granulomatous disease of unknown etiology. Hepatic involvement of sarcoidosis was described in 11.5% of 736 patients enrolled in the ACCESS study. However, presence alone of granulomas in an organ in sarcoidosis does not dictate treatment. The decision to treat should be based on symptoms and severity of disease. Although hepatic involvement usually is asymptomatic, a minority of patients progress to chronic cholestatic disease, portal hypertension, and cirrhosis that may require liver transplantation. Treatment of hepatic sarcoidosis should be reserved for patients who manifest this spectrum of disease. Glucocorticoid treatment is first-line therapy for hepatic sarcoidosis, improving symptoms and abnormal laboratory values but generally having no effect on progression of disease. In addition to glucocorticoids, immunomodulators such as azathioprine, methotrexate, hydroxychloroquine, and infliximab have been used with some positive effects on symptoms, liver enzyme abnormalities, and hepatomegaly, but none has been shown to prevent progression of disease. Ultimately, in cases of overt liver failure, liver transplantation is the definitive treatment. Overall, treatment for hepatic sarcoidosis is targeted toward alleviation of symptoms but has no curative potential at this time. Focus should be on discovering the etiology of the disease to target therapy at prevention, not cure. PMID- 17081482 TI - Liver involvement in cystic fibrosis. AB - The hepatobiliary manifestations of cystic fibrosis (CF) encompass a broad clinical spectrum, from mild steatosis, associated with poor nutrition, to multilobular cirrhosis and the complications of portal hypertension. The factor(s) responsible for the development and progression of liver disease in a subset of patients with CF are unknown. Liver disease can be silent and progressive, manifesting only with complications associated with cirrhosis and portal hypertension. Clinical evaluation for detecting and monitoring the progression of liver disease includes the following: physical examination of the liver, biochemical tests of liver function and injury, and radiological imaging with abdominal ultrasonography. Careful monitoring should take place in all patients with CF, as currently, there are no sensitive and/or specific historical or biochemical markers to predict who is at risk for the development of liver disease. Current treatment options for CF-associated liver disease are very limited. The bile acid ursodeoxycholic acid may improve biochemical parameters of liver disease, but its long-term efficacy in preventing the progression of liver disease in CF is unproven. Treatment therefore rests on optimizing nutritional status; correcting fat-soluble vitamin, essential fatty acid, and other mineral deficiencies; and treating complications of end-stage liver disease, such as pruritus, ascites, and portal hypertension. PMID- 17081483 TI - Treatment of hepatitis C cryoglobulinemia: mission and challenges. AB - Mixed cryoglobulinemia (MC) is a syndrome resulting from cold-insoluble immunoglobulin complexes or cryoglobulins (CGs) that precipitate in the serum of 40% to 50% of patients with chronic hepatitis C virus (HCV) infection. The pathogenesis of cryoglobulinemia likely occurs due to chronic viremia and generation of rheumatoid factor following continuous presentation of antigen immunoglobulin complexes to B cells. CGs are thought to be responsible for a variety of extrahepatic manifestations associated with HCV, including vasculitis, glomerulonephritis, arthritis, and neuropathies, which occur in approximately 10% of HCV patients with CGs. CGs also are a powerful predictive factor for progressive liver disease and the aggressive reoccurrence of liver disease in HCV positive patients after liver transplantation. First-line therapy for MC due to HCV infection is antiviral therapy with pegylated interferon-alpha and ribavirin. Viral eradication usually produces marked reduction of physical complications and arrests end organ damage concomitant with clearance of CG. Additional prospective, controlled studies are necessary to determine whether CG influences patient virologic response and/or its durability to antiviral therapy. Immunomodulators such as corticosteroids and cyclophosphamide are efficacious for palliative treatment of the symptomatology of HCV cryoglobulinemia but may enhance viral replication. Consequently, prolonged therapy with immunomodulatory agents should be limited to severe vasculitis or aggressive glomerulonephritis in patients with MC due to HCV who have failed to respond to antiviral therapy. In acute, fulminant presentations, plasmapheresis may provide temporary relief and arrest the rapid progression of the disease so that additional therapy can be initiated. PMID- 17081484 TI - Psychiatric management of the hepatitis C patient. AB - Patients with hepatitis C virus (HCV) infection have a higher prevalence of psychiatric illness compared with the general US population, and the prevalence of HCV infection in patients with severe mental illness ranges between 8% and 19%, which is four to nine times that of the general US population (1.8%). Given the association between HCV infection and psychiatric illness, gastroenterologists are on the front line of identifying comorbid psychiatric and substance use disorders and conducting a psychosocial pretreatment risk-benefit assessment for HCV infection. The use of interferon-alpha (IFN)-based therapies in combination with ribavirin (RBV) to eradicate HCV has been associated with frequent neuropsychiatric adverse effects (eg, affective, anxiety, cognitive, and psychotic symptoms) that compromise the management of both HCV patients with and those without a preexisting history of psychiatric illness. Consequently, gastroenterologists have been reluctant to engage patients with HCV and comorbid psychiatric illness in antiviral treatment due to concerns about exacerbating or precipitating neuropsychiatric symptoms. Despite the clinical challenge that HCV treatment of patients with comorbid HCV and psychiatric illness presents, recent research indicates that HCV treatments can be safely administered to patients with psychiatric illness provided that there is a comprehensive pretreatment assessment, a risk-benefit analysis, and ongoing follow-up of neuropsychiatric symptoms during antiviral therapy. The process of pretreatment assessment involves screening patients for psychiatric and substance use disorders, educating patients about the treatment process, and addressing available psychosocial support. Most psychotropic medications (antidepressants, mood stabilizers, antipsychotics, and neuroleptics) are thought to be safe to use in the management of patients with HCV and psychiatric illness and for the management of IFN- and RBV-induced neuropsychiatric adverse effects. Nonetheless, the prophylactic use of psychotropic medications to prevent IFN- and RBV-induced neuropsychiatric adverse effects remains a controversial topic. The use of IFN and RBV in patients with HCV and severe mental illness can be done safely with expert psychiatric follow-up. In this review, we discuss the process of pretreatment assessment of patients with HCV and psychiatric illness and specifically address IFN- and RBV-induced depression in patients receiving HCV treatment. PMID- 17081485 TI - Antiviral therapy for hepatitis C in the setting of liver transplantation. AB - Hepatitis C viremia after liver transplantation for hepatitis C virus (HCV) liver disease is universal. Progressive HCV disease after transplantation is the leading cause of death, graft failure, and retransplantation. Whether to treat, with which agents, and timing of therapy are unanswered questions. Timing options include pretransplantation, prophylactic, post-transplantation preemptive, and post-transplantation recurrence-based therapy. The latter is most commonly utilized. There are little data for each of these, much less comparisons. Pegylated interferon-alpha has supplanted standard interferon-alpha due to increased efficacy and is generally used in combination with ribavirin (RBV). Efficacy is less than in nontransplant settings due to immunosuppression, an increased prevalence of genotype 1 HCV, patient comorbidities, and decreased functional status. Administration of HCV therapy to cirrhotic patients prior to transplantation may eradicate or suppress HCV and prevent or reduce severity of recurrence. Sustained virological response (SVR) as high as 50% was attained in genotypes 2 or 3 HCV. Comparison of preemptive and histology-based post transplantation HCV therapy should be done, and more data will be available on pretransplantation therapy. Post-transplant patients are less tolerant of therapy, particularly RBV. SVR, the primary goal of therapy, likely halts disease progression, but only 20% to 30% of treated patients achieve SVR. Preemptive therapy early after transplantation may have advantages due to the potential to delay or blunt severity of graft infection and recurrent hepatitis. In post transplant therapy, RBV toxicity is attenuated in relation to decreased renal function, and side effects of interferon are more prominent. An ongoing trial will assess preemptive therapy with treatment after histologic recurrence. Novel anti-HCV therapies such as protease and polymerase inhibitors are emerging. These must be tested with urgency in the transplant setting. Retransplantation for progressive HCV disease is more controversial due to poor outcomes, graft shortage, and disease recurrence. PMID- 17081486 TI - Treatment of refractory ascites. AB - In 1996, the International Ascites Club defined "refractory ascites" as ascites that cannot be mobilized by medical therapy or that recurs early after initial mobilization despite continued treatment. Of all patients with ascites, 5% to 10% will become refractory to medical therapy. Management of refractory ascites should attempt to control fluid accumulation, reduce the likelihood of developing complications such as spontaneous bacterial peritonitis (SBP) and the hepatorenal syndrome, and improve the patient's nutritional status and overall well-being. Measures to control ascites accumulation include documenting medication and dietary compliance and eliminating potentially nephrotoxic agents that promote sodium retention. Large volume paracentesis is an effective first step in managing these patients and can be performed routinely in an outpatient setting. When more than 5 L of fluid are removed during a paracentesis, intravenous albumin should be infused to reduce the likelihood of the patient developing postparacentesis circulatory dysfunction. Transjugular intrahepatic portosystemic shunt (TIPS) placement effectively eliminates ascites; however, there is no convincing evidence that the shunt improves mortality. Furthermore, it is associated with frequent complications of encephalopathy and shunt malfunction. We feel TIPS should be reserved for patients requiring extremely frequent paracentesis, those who develop significant postparacentesis circulatory dysfunction, or those with hepatic hydrothorax. Patients who have evidence of SBP should be treated with antibiotics and intravenous albumin infusion. Patients who have had a previous episode of SBP or an ascitic fluid protein level of less than 1.0 should receive prophylactic antibiotics. Overall, the prognosis for patients with refractory ascites remains grim, and liver transplantation is the only definitive therapy. Appropriate candidates should be identified promptly and referred for transplant evaluation. PMID- 17081487 TI - Empirical Bayes before-after safety studies: lessons learned from two decades of experience and future directions. AB - The empirical Bayes (EB) methodology has been applied for over 20 years now in conducting statistically defendable before-after studies of the safety effect of treatments applied to roadway sites. The appeal of the methodology is that it corrects for regression to the mean and traffic volume and other changes not due to the measure. There is, therefore, a natural tendency to put a stamp of approval on any study that uses this methodology, and to assume that the results can then be used in specifying crash modification factors for use in developing treatments for hazardous locations, or in designing new roads using tools such as the interactive highway safety design model (IHSDM). At the other extreme are skeptics who suggest that the increased sophistication and data needs of the EB methodology are not worth the effort since alternative, less complex methods can produce equally valid results. The primary objective of this paper is to capitalize on experience gained from two decades of conducting EB studies around the world to illustrate that the EB methodology, if properly undertaken, produces results that could be substantially different and less biased than those from more conventional types of studies. A secondary objective is to emphasize that caution is needed in assessing the validity of studies undertaken with the EB methodology and in using these results for providing crash modification factors. To this end, a number of issues that are critical to the proper conduct and interpretation of EB evaluations are raised and illustrated based on lessons learned from recent experience with these studies. These include: amalgamating the effects on different crash types; the specification of the reference/comparison groups; and accounting for traffic volume changes. Current and future directions, including the improvements offered by a full Bayes approach, are discussed. PMID- 17081488 TI - The 'like me' framework for recognizing and becoming an intentional agent. AB - Infant imitation demonstrates that the perception and production of human action are closely linked by a 'supramodal' representation of action. This action representation unites observation and execution into a common framework, and it has far-reaching implications for the development of social cognition. It allows infants to see the behaviors of others as commensurate with their own-as 'like me.' Based on the 'like me' perception of others, social encounters are interpretable and informative. Infants can use themselves as a framework for understanding others and can learn about the possibilities and consequences of their own potential acts by observing the behavior of others. Through social interaction with other intentional agents who are viewed as 'like me,' infants develop a richer social cognition. This paper explores the early manifestations and cascading developmental effects of the 'like me' conception. PMID- 17081489 TI - 'Obsessed with goals': functions and mechanisms of teleological interpretation of actions in humans. AB - Humans show a strong and early inclination to interpret observed behaviours of others as goal-directed actions. We identify two main epistemic functions that this 'teleological obsession' serves: on-line prediction and social learning. We show how teleological action interpretations can serve these functions by drawing on two kinds of inference ('action-to-goal' or 'goal-to-action'), and argue that both types of teleological inference constitute inverse problems that can only be solved by further assumptions. We pinpoint the assumptions that the three currently proposed mechanisms of goal attribution (action-effect associations, simulation procedures, and teleological reasoning) imply, and contrast them with the functions they are supposed to fulfil. We argue that while action-effect associations and simulation procedures are generally well suited to serve on-line action monitoring and prediction, social learning of new means actions and artefact functions requires the inferential productivity of teleological reasoning. PMID- 17081490 TI - Variability in the immunodetection of His-tagged recombinant proteins. AB - Labeling of recombinant proteins with polypeptide fusion partners, or affinity tagging, is a useful method to facilitate subsequent protein purification and detection. Poly-histidine tags (His-tags) are among the most commonly used affinity tags. We report strikingly variable immunodetection of two His-tagged recombinant human erythropoietins (Epo): wild type Epo (Epo(wt)) and Epo containing an R103A mutation (Epo(R103A)). Both were engineered to contain a C terminal six residue His-tag. The cDNA constructs were stably transfected into Chinese hamster ovary (CHO) cells and COS-7 cells. Clones from the CHO cell transfections were selected for further characterization and larger-scale protein expression. Three chromatographic steps were utilized to achieve pharmacologically pure Epo. Conditioned media from the Epo-expressing cell lines and protein-containing samples from each step of purification were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and dot blot, using both monoclonal anti-human Epo antibody (AE7A5) and anti-His antibodies. While the successful incorporation of the His-tag into our constructs was confirmed by Epo binding to Ni(2+)- nitrilotriacetic acid resin and by microcapillary reverse phase high-performance liquid chromatography nano-electrospray tandem mass spectrometery amino acid sequencing, the levels of immunodetection of His-tagged protein varied markedly depending on the particular anti-His-tag antibody used. Such variability in His-tag immunorecognition can lead to critical adverse effects on several analytical methods. PMID- 17081491 TI - Alzheimer's-disease-associated conformation of intrinsically disordered tau protein studied by intrinsically disordered protein liquid-phase competitive enzyme-linked immunosorbent assay. AB - Tau protein, the major constituent of paired helical filaments in Alzheimer's disease, belongs to the intrinsically disordered proteins (IDPs). IDPs are an emerging group in the protein kingdom characterized by the absence of a rigid three-dimensional structure. Disordered proteins usually acquire a "functional fold" upon binding to their interaction partner(s). This property of IDPs implies the need for innovative approaches to measure their binding affinity. We have mapped and measured the Alzheimer's-disease-associated epitope on intrinsically disordered tau protein with a novel two-step sandwich competitive enzyme-linked immunosorbent assay (ELISA). This approach allowed us to determine the binding affinity of disordered tau protein in liquid phase without any disturbance to the competitive equilibrium and without any need for covalent or noncovalent modification of tau protein. Furthermore, the global fitting method, used for the reconstruction of tau binding curves, significantly improved the assay readout. The proposed novel competitive ELISA allowed us to determine the changes in the standard Gibbs energy of binding, thus enabling measurement of tau protein conformation in the core of paired helical filaments. IDP competitive ELISA results showed, for the first time, that the tau protein C terminus of the Alzheimer's-disease-derived paired helical filaments core subunit adopts beta turn type I' fold and is accessible from solution. PMID- 17081492 TI - Hydroxyl-specific fluorescence labeling of ABP-deoxyguanosine, PhIP deoxyguanosine, and AFB1-formamidopyrimidine with BODIPY-FL. AB - Detection and analysis of DNA adducts resulting from endogenous or exogenous exposures to carcinogens are essential not only for quantifying biologically effective doses but also for establishing relationships between exposure and cancer risk. We have developed and validated a procedure of high sensitivity and specificity based on fluorescence labeling of DNA adducts combined with high performance liquid chromatography-laser-induced fluorescence detection. The fluorescent dye 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3 propionic acid (BODIPY FL) was used to label the deoxynucleoside adducts N-(2' deoxyguanosine-8-yl)-4-aminobiphenyl and N-(2'-deoxyguanosine-8-yl)-2-amino-1 methyl-6-phenylimidazo[4,5-b]pyridine and the base adduct aflatoxin B(1) formamidopyrimidine by acylation. The labeling reaction was carried out on adducts at 1pmol to 30nmol concentrations at 25 degrees C for 4h in dichloromethane with 200- to 5000-fold excess of BODIPY FL. BODIPY FL and its activating agents 1,3-dicyclohexylcarbodiimide and 4-dimethylaminopyridine were used at a molar ratio of 1:2:2. Under these conditions, all of the above adducts were quantitatively converted to bis-labeled products, as confirmed by mass spectrometry. Sites of derivatization of adduct deoxynucleosides were established primarily by nuclear magnetic resonance and by collision-induced dissociation mass spectrometric analysis, which indicated that the bis-BODIPY groups were located predominantely on the 3'- and 5'-hydroxyl groups of the deoxyribose ring. PMID- 17081493 TI - Enzymatic assay of allantoin in serum using allantoinase and allantoate amidohydrolase. AB - A new enzymatic assay for specifically measuring allantoin concentration in serum has been developed. The currently used methods for allantoin analysis are time consuming and nonspecific or depend on the use of expensive equipment. In our method, allantoin is converted to allantoate by the action of allantoinase (EC 3.5.2.5). The allantoate produced is hydrolyzed to ureidoglycine and ammonia by the action of allantoate amidohydrolase (EC 3.5.3.9). Nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase (EC 1.4.1.4) subsequently acts on the ammonia produced, resulting in a change in absorbance at 340nm due to the consumption of reduced nicotinamide adenine dinucleotide phosphate. The amount of allantoin present is related to the change in the absorbance. The standard curve is linear up to at least 1mM allantoin. The procedure is simple, rapid, and accurate. The method has been used to measure serum allantoin levels after oral administration of purine nucleotides to experimental animals, including rats that have uricase catalyzing the conversion of urate to allantoin. PMID- 17081494 TI - The cytochromes P450 (CYP) response to allergic inflammation of the lung. AB - The expression of the mouse Cyp family and key inflammatory mediators were examined in a model of ovalbumin (OVA)-induced allergic airway disease. The expression of IL-4, IL-13 and Ccl11 increased during the acute phase of allergic inflammation and decreased with its resolution. Interestingly, the expression of Ccl20 was increased during the resolution phase. The response of the Cyp gene family to the development of allergic inflammation was differential and correlated with the evolution of the inflammatory response. During the acute inflammatory phase the mRNA levels of Cyp2e1, Cyp2f2, Cyp2j6, Cyp4b1, Cyp8a1 and Cypor were decreased while the mRNA levels of Cyp4f18, Cyp5a1 and Cyp7b1 were elevated. With resolution of the inflammation the expression patterns returned to normal. These changes suggest that the Cyp family may play a role in the allergic inflammation by modulating the metabolism of xenobiotics and endogenous compounds such as LTB4, TXA1, PGI2 and native anti-glucocorticoids. PMID- 17081495 TI - Pulse radiolysis and steady-state analyses of the reaction between hydroethidine and superoxide and other oxidants. AB - Hydroethidine (HE) is a cell-permeable probe used for the intracellular detection of superoxide. Here, we report the direct measurement of the rate constant between hydroethidine and superoxide radical anion using the pulse radiolysis technique. This reaction rate constant was calculated to be ca. 2 x 10(6) M(-1) s(-1) in water:ethanol (1:1) mixture. The spectral characteristics of the intermediates indicated that the one-electron oxidation product of HE was different from the one-electron reduction product of ethidium (E+). The HPLC electrochemical measurements of incubation mixtures containing HE and the oxygenated Fenton's reagent (Fe2+/DTPA/H2O2) in the presence of aliphatic alcohols or formate as a superoxide generating system revealed 2-OH-E+ as a major product. Formation of 2-OH-E+ by the Fenton's reagent without additives was shown to be superoxide dismutase-sensitive and we attribute the formation of superoxide radical anion to the one-electron reduction of oxygen by the DTPA-derived radical. Addition of tert-butanol, DMSO, and potassium bromide to the Fenton's system caused inhibition of 2-OH-E+ formation. Results indicate that reducing and oxidizing radicals have differential effects on the formation of 2-OH-E+. PMID- 17081496 TI - Impact of GPCRs in clinical medicine: monogenic diseases, genetic variants and drug targets. AB - By virtue of their large number, widespread distribution and important roles in cell physiology and biochemistry, G-protein-coupled receptors (GPCR) play multiple important roles in clinical medicine. Here, we focus on 3 areas that subsume much of the recent work in this aspect of GPCR biology: (1) monogenic diseases of GPCR; (2) genetic variants of GPCR; and (3) clinically useful pharmacological agonists and antagonists of GPCR. Diseases involving mutations of GPCR are rare, occurring in <1/1000 people, but disorders in which antibodies are directed against GPCR are more common. Genetic variants, especially single nucleotide polymorphisms (SNPs), show substantial heterogeneity in frequency among different GPCRs but have not been evaluated for some GPCR. Many therapeutic agonists and antagonists target GPCR and show inter-subject variability in terms of efficacy and toxicity. For most of those agents, it remains an open question whether genetic variation in primary sequence of the GPCR is an important contributor to such inter-subject variability, although this is an active area of investigation. PMID- 17081497 TI - Cloning, expression, and characterisation of a Plasmodium vivax MSP7 family merozoite surface protein. AB - Plasmodium vivax remains the most widespread Plasmodium parasite species around the world, producing about 75 million malaria cases, mainly in South America and Asia. A vaccine against this disease is of urgent need, making the identification of new antigens involved in target cell invasion, and thus potential vaccine candidates, a priority. A protein belonging to the P. vivax merozoite surface protein 7 (PvMSP7) family was identified in this study. This protein (named PvMSP7(1)) has 311 amino acids displaying an N-terminal region sharing high identity with P. falciparum MSP7, as well as a similar proteolytical cleavage pattern. This protein's expression in P. vivax asexual blood stages was revealed by immuno-histochemical and molecular techniques. PMID- 17081498 TI - Processing of VSVG protein is not a rate-limiting step for its efflux from the Golgi complex. AB - The secretory membrane system is comprised of membrane-bound organelles defined by specific sets of proteins that function in sequential modification of cargo proteins and lipids. This processing of cargo proteins and lipids is coupled to their secretory transport. Here, we investigated the effect of inhibiting N glycan processing by swainsonine, an inhibitor of Golgi alpha1,2-mannosidase-II, on secretory transport of the thermo-reversible tsO45 mutant of vesicular stomatitis virus glycoprotein tagged with green fluorescent protein (VSVG-FP). Quantitative analysis using kinetic modeling combined with live cell imaging was used to derive the rate coefficients that delineate secretory transport of VSVG FP. We found that neither inhibition of N-glycan processing nor elimination by mutagenesis of the first of the two asparagine-linked glycans had any significant effect on the rate of VSVG-FP transport through the Golgi. These data suggest that at least for VSVG, the multi-enzymatic process of N-glycan modification does not comprise a rate-limiting step for its Golgi efflux. PMID- 17081499 TI - Small heat shock proteins protect against alpha-synuclein-induced toxicity and aggregation. AB - Protein misfolding and inclusion formation are common events in neurodegenerative diseases, such as Parkinson's disease (PD), Alzheimer's disease (AD) or Huntington's disease (HD). Alpha-synuclein (aSyn) is the main protein component of inclusions called Lewy bodies (LB) which are pathognomic of PD, Dementia with Lewy bodies (DLB), and other diseases collectively known as LB diseases. Heat shock proteins (HSPs) are one class of the cellular quality control system that mediate protein folding, remodeling, and even disaggregation. Here, we investigated the role of the small heat shock proteins Hsp27 and alphaB crystallin, in LB diseases. We demonstrate, via quantitative PCR, that Hsp27 messenger RNA levels are approximately 2-3-fold higher in DLB cases compared to control. We also show a corresponding increase in Hsp27 protein levels. Furthermore, we found that Hsp27 reduces aSyn-induced toxicity by approximately 80% in a culture model while alphaB-crystallin reduces toxicity by approximately 20%. In addition, intracellular inclusions were immunopositive for endogenous Hsp27, and overexpression of this protein reduced aSyn aggregation in a cell culture model. PMID- 17081500 TI - UT-B1 urea transporter plays a noble role as active water transporter in C6 glial cells. AB - Since our experimental results suggest that UT-B1 functions as active water transporter against osmotic gradient in C6 glial cells, we report here for the first time the evidence for the active water transport. Exposure of C6 cells to a hyperosmotic solution containing glycerol or sucrose produced cell shrinkage due to water efflux according to osmotic gradient for water movement. On the other hand, C6 cells show cell swelling against osmotic gradient for water movement just after exposure to a hyperosmotic solution containing urea, indicating that water influx against osmotic gradient for water movement is accelerated by urea; i.e., urea performs active water transport. A specific inhibitor of UT-B, pCMBS, blocked the urea-induced swelling. The urea-induced cell swelling was significantly suppressed in the siRNA-induced UT-B1-knockdown C6 cells. Taken together, these observations indicate that UT-B1 acts as an active water transporter, providing a new model on active water transport. PMID- 17081501 TI - Same causes, same cures. AB - Thanks to the continuing bio-medicinal efforts, similar causes underlying the pathogenesis of Alzheimer's disease (AD) and prion diseases (PDs) have been revealed, which include oxidative stress, excessive transition metal ions, and misfolded/aggregated proteins. Therefore, the therapeutic strategy for one disease may be effective for the other. More interestingly, accumulating evidence indicates that not just the strategies but also the prescriptions may be shared by AD and PD treatments. In this review, we first summarize the known dual fighters against AD and PDs (which include antioxidants, metal chelators, and protein aggregation inhibitors), and then indicate that some super-dual-fighters may hit multiple targets implicated in AD and PDs, whose structural features highlight the importance of aromatic moiety and phenolic groups. These findings not only provide important clues to accelerating the screening of anti-AD and anti-PDs drugs but also help to understand the etiology of AD and PDs. PMID- 17081502 TI - Properties of flavonoids influencing the binding to bilitranslocase investigated by neural network modelling. AB - Bilitranslocase is a plasma membrane carrier firstly identified on the sinusoidal (vascular) domain of liver cells and later on also in the gastric epithelium. It transports diverse organic anions, such as bilirubin, some phthaleins and many dietary anthocyanins, suggesting that it could play a role both in the absorption of flavonoids from dietary sources and in their hepatic metabolism. This work was aimed at characterising the interaction of bilitranslocase with flavonols, a flavonoid sub-class. The results obtained show that, contrary to anthocyanins, flavonol glycosides do not interact with the carrier, whereas just some of the corresponding aglycones act as relatively poor ligands to bilitranslocase. These data point to a clear-cut discrimination between anthocyanins and flavonols occurring at the level of the bilitranslocase transport site. A quantitative structure-activity relationship based on counter propagation artificial neural network modelling was undertaken in order to shed light on the nature of flavonoid interaction with bilitranslocase. It was found that binding relies on the ability to establish hydrogen bonds, ruling out the involvement of charge interactions. This requisite might be at the basis of the discrimination between anthocyanins and flavonols by bilitranslocase and could lie behind some aspects of the distinct pharmacokinetic properties of anthocyanins and flavonols in mammals. PMID- 17081503 TI - Mechanism of action of AZD0865, a K+-competitive inhibitor of gastric H+,K+ ATPase. AB - AZD0865 is a member of a drug class that inhibits gastric H(+),K(+)-ATPase by K(+)-competitive binding. The objective of these experiments was to characterize the mechanism of action, selectivity and inhibitory potency of AZD0865 in vitro. In porcine ion-leaky vesicles at pH 7.4, AZD0865 concentration-dependently inhibited K(+)-stimulated H(+),K(+)-ATPase activity (IC(50) 1.0+/-0.2 microM) but was more potent at pH 6.4 (IC(50) 0.13+/-0.01 microM). The IC(50) values for a permanent cation analogue, AR-H070091, were 11+/-1.2 microM at pH 7.4 and 16+/ 1.8 microM at pH 6.4. These results suggest that the protonated form of AZD0865 inhibits H(+),K(+)-ATPase. In ion-tight vesicles, AZD0865 inhibited H(+),K(+) ATPase more potently (IC(50) 6.9+/-0.4 nM) than in ion-leaky vesicles, suggesting a luminal site of action. AZD0865 inhibited acid formation in histamine- or dibutyryl-cAMP-stimulated rabbit gastric glands (IC(50) 0.28+/-0.01 and 0.26+/ 0.003 microM, respectively). In ion-leaky vesicles at pH 7.4, AZD0865 (3 microM) immediately inhibited H(+),K(+)-ATPase activity by 88+/-1%. Immediately after a 10-fold dilution H(+),K(+)-ATPase inhibition was 41%, indicating reversible binding of AZD0865 to gastric H(+),K(+)-ATPase. In contrast to omeprazole, AZD0865 inhibited H(+),K(+)-ATPase activity in a K(+)-competitive manner (K(i) 46+/-3 nM). AZD0865 inhibited the process of cation occlusion concentration dependently (IC(50) 1.7+/-0.06 microM). At 100 microM, AZD0865 reduced porcine renal Na(+),K(+)-ATPase activity by 9+/-2%, demonstrating a high selectivity for H(+),K(+)-ATPase. Thus, AZD0865 potently, K(+)-competitively, and selectively inhibits gastric H(+),K(+)-ATPase activity and acid formation in vitro, with a fast onset of effect. PMID- 17081504 TI - Genomewide linkage scan for nicotine dependence: identification of a chromosome 5 risk locus. AB - BACKGROUND: Nicotine dependence (ND) is costly to societies worldwide, moderately heritable, and genetically complex. Risk loci can be identified with genetic linkage analysis independent of prior physiological hypotheses. METHODS: We completed a genomewide linkage scan to map loci increasing risk for DSM-IV ND and for a quantitative assessment of ND as measured by the Fagerstrom Test for Nicotine Dependence (FTND) in a set of 634 small nuclear families ascertained on the basis of multiple individuals affected with cocaine or opioid dependence. Of these, 507 had at least two subjects affected with ND. There are two distinct populations within this sample, European-Americans (EAs) and African-Americans (AAs). RESULTS: A region on chromosome 5 was identified as containing a gene that affects risk for ND on the basis of FTND score in the AA part of our sample (logarithm of the odds [lod] score 3.04; empirically determined to be genomewide significant, p = .0374; point p = .0001). The highest lod score observed in the EA part of the sample was on chromosome 7 (lod score 2.73). Several other "possible" risk loci were identified in either AA or EA subjects, with many of these in proximity to previously suggested risk loci from other clinical samples. Three nominally significant single-nucleotide polymorphism associations were found at the peptidylglycine alpha-amidating monooxygenase (PAM) locus under the chromosome 5 linkage peak, also in the AA part of the sample. CONCLUSIONS: These data add to the growing evidence for locations for ND risk loci, add a novel statistically significant locus important in AAs, and suggest a gene that might be contributing to this linkage signal. PMID- 17081506 TI - Differential effects of the stimulus sequence on CNV and P300. AB - During the presentation of stimulus sequences in oddball paradigms, participants tend to implicitly evaluate the conditional probability of target occurrence. It is not sure, however, if subjective estimation of conditional probabilities modulates target expectancy and target processing in the same manner. In the present experiment, the amplitudes of CNV and P300 were studied separately to compare preparatory and decision mechanisms and their sensitivity to variations in target probability. Amplitudes of both components were measured at different positions in the stimulus sequence, which modulates target probability as a function of distance from the preceding target. Results showed shorter RTs with increased probability of target occurrence (that is, with longer distance to the previous target). CNV amplitude was low when target probability was zero and maximal when the occurrence of a target was possible, regardless of its probability. Conversely, a gradual increase with augmenting probability was observed for P300 amplitude. Thus, preparatory activity as reflected by the CNV showed an all or nothing response to variations in probability, whereas action closure mechanisms, indexed by P300 components, were found to be more sensitive to subtle differences in stimulus expectancy. PMID- 17081505 TI - The striatal-enriched protein tyrosine phosphatase gates long-term potentiation and fear memory in the lateral amygdala. AB - BACKGROUND: Formation of long-term memories is critically dependent on extracellular-regulated kinase (ERK) signaling. Activation of the ERK pathway by the sequential recruitment of mitogen-activated protein kinases is well understood. In contrast, the proteins that inactivate this pathway are not as well characterized. METHODS: Here we tested the hypothesis that the brain specific striatal-enriched protein tyrosine phosphatase (STEP) plays a key role in neuroplasticity and fear memory formation by its ability to regulate ERK1/2 activation. RESULTS: STEP co-localizes with the ERKs within neurons of the lateral amygdala. A substrate-trapping STEP protein binds to the ERKs and prevents their nuclear translocation after glutamate stimulation in primary cell cultures. Administration of TAT-STEP into the lateral amygdala (LA) disrupts long term potentiation (LTP) and selectively disrupts fear memory consolidation. Fear conditioning induces a biphasic activation of ERK1/2 in the LA with an initial activation within 5 minutes of training, a return to baseline levels by 15 minutes, and an increase again at 1 hour. In addition, fear conditioning results in the de novo translation of STEP. Inhibitors of ERK1/2 activation or of protein translation block the synthesis of STEP within the LA after fear conditioning. CONCLUSIONS: Together, these data imply a role for STEP in experience-dependent plasticity and suggest that STEP modulates the activation of ERK1/2 during amygdala-dependent memory formation. The regulation of emotional memory by modulating STEP activity may represent a target for the treatment of psychiatric disorders such as posttraumatic stress disorder (PTSD), panic, and anxiety disorders. PMID- 17081507 TI - Structural studies of the O-antigenic polysaccharides from Shigella dysenteriae type 3 and Escherichia coli O124, a reinvestigation. AB - The structures of the O-antigenic part of the lipopolysaccharides from Shigella dysenteriae type 3 and Escherichia coli O124 have been reinvestigated. (1)H and (13)C NMR spectroscopy in combination with selected 2D NMR techniques were used to determine the O-antigen pentasaccharide repeating units with the following structure: [see text]. From biosynthetic considerations this should also be the biological repeating unit. The structures of the repeating units also explain the previously observed cross-reactivity between the strains and to E. coli O164, which only differs in the terminal sugar residue that is lacking the (R)-1 carboxyethyl group. PMID- 17081508 TI - Structure and serological analysis of the Hafnia alvei 481-L O-specific polysaccharide containing phosphate in the backbone chain. AB - The lipopolysaccharide was extracted from cells of Hafnia alvei 481-L bacterial strain and, after mild acid hydrolysis, the O-specific polysaccharide was isolated and characterised. On the basis of chemical analyses and NMR spectroscopic studies of the polysaccharide and oligosaccharides obtained after Smith degradation, or hydrogen fluoride treatment, it was found that the repeating unit of the O-specific polysaccharide is a phosphorylated hexasaccharide: [see text]. The biological repeating unit of the H. alvei 481-L O antigen has galactose phosphate at the nonreducing terminus. Serological tests indicate that this strain represents an individual serotype in the H. alvei genus. PMID- 17081509 TI - Evidence of the presence of 2-O-beta-D-xylopyranosyl-alpha-l-arabinofuranose side chains in barley husk arabinoxylan. AB - (Glucurono)arabinoxylans were extracted from barley husks and degraded with endo beta-xylanase or subjected to periodate oxidation. The released oligosaccharide fragments were separated and isolated on Biogel-P2, and their structures were determined by NMR spectroscopy. The oligosaccharides identified consisted of beta d-(1-->4)-linked xylopyranosyl residues, of which some were substituted at O-3 with alpha-l-arabinofuranosyl groups or at O-2 with 4-O-methylglucuronic acid. In addition to these substituents, a disaccharide side chain, 2-O-beta-d xylopyranosyl-alpha-l-arabinofuranose, attached at position O-3 of the main chain, was proved to exist in arabinoxylan from barley husks. The compound was fully characterized with NMR, and all (1)H and (13)C NMR signals were assigned. The arabinose to xylose ratio was low (approximately 0.2) and no 2,3 disubstitution existed. No blocks of substituted xylose residues could be observed along the main chain. PMID- 17081510 TI - Acidic cesium salts of 12-tungstophosphoric acid as catalysts for the dehydration of xylose into furfural. AB - Cesium salts of 12-tungstophosphoric acid, Cs(x)H(3-x)PW(12)O(40) (Cs(x)PW), in the bulk form or supported on medium-pore MCM-41 (3.7 nm) or large-pore (9.6 nm) micelle-templated silicas are active solid acid catalysts for the cyclodehydration of xylose into furfural, in a toluene/water solvent system (T/W) or in dimethyl sulfoxide (DMSO). The catalytic results are comparable to those obtained using sulfuric acid, under similar reaction conditions. The initial activities increase in the order H(3)PW(12)O(40)40% and decreased ECM-mediated phosphorylation of FAK Y397 and ERK1/2. Serine phosphorylation of Runx2/Cbfa-1 was significantly reduced after 8 days in treated cells. Finally, FAK inhibition blocked osterix transcriptional activity and the osteogenic differentiation of hMSC, as assessed by lowered expression of osteogenic genes (RT-PCR), decreased alkaline phosphatase activity, greatly reduced calcium deposition, and a lower mineral:matrix ratio after 28 days in culture. These results suggest that FAK signaling plays an important role in regulating ECM-induced osteogenic differentiation of hMSC. PMID- 17081518 TI - Plasmalogens in the retina: in situ hybridization of dihydroxyacetone phosphate acyltransferase (DHAP-AT)--the first enzyme involved in their biosynthesis--and comparative study of retinal and retinal pigment epithelial lipid composition. AB - Plasmalogens (Pls) are phospholipids containing a vinyl-ether bond in the sn-1 position of the glycerol backbone. The physiological role of Pls is still enigmatic, especially within the eye where their deficiency leads to developmental abnormalities. In order to learn more about the functions of Pls in the posterior eye, we evaluated retinal Pl content as well as the expression of the first enzyme involved in Pls biosynthesis, dihydroxyacetone phosphate acyltransferase (DHAP-AT) in the retina. In situ hybridization of DHAP-AT mRNA was performed on rat eye sections. The Pl contents of calf retina and retinal pigment epithelium (RPE) samples were determined by high-performance liquid chromatography, thin-layer chromatography, and gas chromatography. DHAP-AT was highly expressed in the inner segment of photoreceptors and in the RPE, suggesting two distinct sites for Pl biosynthesis. Plasmenyl-ethanolamine was the prominent class of Pls in both neural retina and RPE (28-29% of the total phospho ethanolamine-glycerides). According to the nature of the alkenyl residue linked to the sn-1 position of Pls, the most striking finding was the greater proportion of octadecanal-aldehyde in the sn-1 position of plasmenyl-ethanolamine of the neural retina compared to all the other classes of Pls in the neural retina and the RPE. These findings might be relevant to the biological functions of Pls against oxidative stress and in the formation of lipid rafts. PMID- 17081519 TI - cAMP-responsive element binding protein mediates a cGMP/protein kinase G dependent anti-apoptotic signal induced by nitric oxide in retinal neuro-glial progenitor cells. AB - Nitric oxide (NO) is cytoprotective to certain types of neuronal cells. The neuroprotective ability of NO in the retina was reportedly mediated by the cyclic GMP (cGMP) to protein kinase G (PKG) pathway. Cyclic AMP-responsive element binding protein (CREB) plays an essential role in the NO/cGMP/PKG-mediated survival of rat cerebellar granule cells. We tested whether CREB transduces the NO/cGMP/PKG anti-apoptotic cascade in R28 neuro-glial progenitor cells. Apoptosis was induced in R28 cells by serum deprivation for 24 h. Varying concentrations of two NO donors, sodium nitroprusside (SNP) and nipradilol, were added to medium with or without an NO scavenger, a soluble guanylyl cyclase inhibitor, or a PKG inhibitor. The cells were immunostained against activated caspase-3 and counterstained with Hoechst 33258. Apoptosis was quantified by counting activated caspase-3 positive or pyknotic cells. SNP and nipradilol rescued R28 cells from apoptosis in a dose-dependent manner, at an optimal concentration of 1.0 microM and 10 microM, respectively. Higher concentrations were cytotoxic. The NO scavenger and the inhibitors decreased the anti-apoptotic effect of the NO donors. Intracellular cGMP levels were increased after exposure to SNP and nipradilol. Western blotting showed that both NO donors increased CREB phosphorylation, which was blocked when pre-exposed to the inhibitors. Transfection with a dominant negative CREB construct defective of phosphorylation at Ser-133 interfered with the anti-apoptotic activity of SNP. These results indicate that CREB at least in part mediates the cGMP/PKG-dependent anti apoptotic signal induced by NO in R28 cells. PMID- 17081520 TI - Activation of transcription factor c-jun in dorsal root ganglia induces VIP and NPY upregulation and contributes to the pathogenesis of neuropathic pain. AB - Vasoactive intestinal peptide (VIP) and neuropeptide Y (NPY) in dorsal root ganglia (DRGs) are known to be upregulated and to contribute to the mechanisms of neuropathic pain following peripheral nerve injury. Moreover, transcription factor c-Jun regulates the expressions of both VIP and NPY in cultured DRG neurons. To elucidate the role of c-Jun in the induction of neuropathic pain hypersensitivity, we examined whether activated c-Jun affects pain behavior and the expressions of VIP and NPY following chronic constriction injury (CCI) of rat sciatic nerve. Intrathecal treatment with c-jun antisense oligodeoxynucleotides (AS-ODN) significantly reduced mechanical allodynia, but not thermal hyperalgesia following CCI. In addition, c-jun AS-ODN also suppressed the remarkable elevations of VIP and NPY mRNAs and the percentages of phosphorylated c-Jun-, VIP , and NPY-immunoreactive neurons observed in DRGs following CCI. These results show that the activation of c-Jun in DRGs induces VIP and NPY upregulation and contributes to the pathogenesis of neuropathic pain following CCI. PMID- 17081521 TI - Fertilization and pregnancy outcome after intracytoplasmic injection with fresh or cryopreserved ejaculated spermatozoa. AB - OBJECTIVE: To assess fertilization, implantation, and pregnancy rates in patients undergoing ICSI using fresh and cryopreserved sperm from ejaculated semen samples. DESIGN: Retrospective study. SETTING: Academic and private medical centers. PATIENT(S): One hundred fifty-eight patients. INTERVENTION(S): Intracytoplasmic sperm injection. MAIN OUTCOME MEASURE(S): Fertilization, implantation, and pregnancy rates were evaluated; 61 cycles of ICSI were performed with cryopreserved and 79 cycles of ICSI were performed with fresh spermatozoa. Also, we divided the outcomes according to the semen characteristics, normozoospermia, oligozoospermia, asthenozoospermia, and oligoasthenozoospermia. RESULT(S): Overall, normal-fertilization rates were higher using fresh sperm (73.8%) compared with cryopreserved sperm (68.7%). Cycles performed in patients with normozoospermia or oligozoospermia had similar fertilization, implantation, and pregnancy rates using fresh or cryopreserved sperm. When asthenozoospermic and oligoasthenozoospermic semen samples were used, the normal-fertilization rate was higher with fresh sperm compared with cryopreserved sperm. However, implantation and pregnancy rates were similar in fresh and cryopreserved sperm samples from patients with asthenozoospermia or oligoasthenozoospermia. CONCLUSION(S): Semen with abnormalities in the motility may be more susceptible to sperm cryopreservation damage, resulting in lower fertilization rates. However, once the oocyte is fertilized, implantation and pregnancy rates are similar to those in patients with oligozoospermia and normozoospermia. PMID- 17081522 TI - Relationship between hypoactive sexual desire disorder and aging. AB - OBJECTIVE: Explore the association between Hypoactive Sexual Desire Disorder (HSDD) and aging. The American Foundation of Urologic Disease and the American Psychiatric Association stipulate that HSDD is only diagnosed when both low sexual desire and sexually related personal distress are present. DESIGN: Community-based, cross-sectional study. SETTING: Europe (UK, Germany, France, Italy) and the USA. PATIENT(S): Women aged 20-70 in sexual relationships participating in the Women's International Study of Health and Sexuality (n=1998 Europe, n=1591 USA). INTERVENTION(S): No interventions were administered. MAIN OUTCOME MEASURES: Self-administered questionnaire that included two validated instruments: Profile of Female Sexual Function(c) measured sexual desire; Personal Distress Scale(c) measured sexual distress. Women with low desire and distress were considered to have HSDD. RESULTS: The proportion of European women with low desire increased from 11% amongst women aged 20-29 years to 53% amongst women aged 60-70 years. The proportion of American women with low desire displayed a trend towards an increase with age. In the 20-29 year age group 65% of European women and 67% of American women with low sexual desire were distressed by it. This decreased to 22% and 37%, respectively, in the 60-70 year age group. In Europe and the USA the prevalence of HSDD in the population did not change significantly with age (6-13% in Europe, 12-19% in the USA). CONCLUSIONS: The proportion of women with low desire increased with age while the proportion of women distressed about their low desire decreased with age. Consequently, the prevalence of HSDD remained essentially constant with age. This may explain why no association between HSDD and age is often reported in the literature. PMID- 17081523 TI - Effect of reduced oxygen concentrations on the outcome of in vitro fertilization. AB - We compared the effects of two standard oxygen concentrations, physiological (5% O(2), 5% CO(2), and 90% N(2)) and atmospheric (5% CO(2) with the balance as air), on fertilization, embryo development, and pregnancy rate in 106 patients undergoing IVF, excluding donor oocyte cycles and preimplantation genetic diagnosis cycles. The differences in oxygen concentration did not significantly affect fertilization rate, blastocyst formation, or pregnancy rate, but there was a significant difference in mean embryo score between physiological and atmospheric groups on day 3. PMID- 17081524 TI - Acceptance of single-embryo transfer by patients. AB - In New Zealand, patient motivation to accept single-embryo transfer can be measurably changed within a short period of time if patients are provided with credible clinical written and verbal communication, in addition to funding incentives from the Ministry of Health. PMID- 17081525 TI - Testosterone replacement increases vasodilatory reserve in androgen-deficient female rats. AB - Testosterone replacement leads to increased coronary pressure-induced tone and vasodilatory reserve in oophorectomized rats. The mechanisms involve changes in endothelial nitric-oxide modulation of smooth-muscle constriction and are not a result of the peripheral conversion of T to estrogen. PMID- 17081526 TI - Automation is the key to standardized semen analysis using the automated SQA-V sperm quality analyzer. AB - OBJECTIVE: To evaluate the performance of the automated semen quality analyzer system for assessing sperm quality. DESIGN: Double-blind prospective study. SETTING: Tertiary care hospital. PATIENT(S): Fifty healthy men donated semen samples. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Precision, accuracy and agreement between automated and manual semen analysis methods was assessed for sperm concentration, motility, morphology, and known concentrations of latex bead quality control media. RESULT(S): A good agreement was seen between the results of sperm concentration reported by the SQA-V automated analyzer (Spermalite/SQA V; Medical Electronic Systems Ltd, Caesarea Industrial Park, Israel) and those obtained manually. A similar linearity was seen when the SQA-V results were compared with the manual data and also when the manual results of individual operators were compared with each other. The automated assessment of morphology showed high sensitivity (89.9%) for identifying percent normal morphology, and the precision of the SQA-V was considerably higher when compared with the manual method. The interoperator variability for manual assessment was significant. The automated analysis was quick compared with the manual method. CONCLUSION(S): The SQA-V can be used interchangeably with manual semen analysis methods for examining sperm concentration and motility. The automated SQA-V analyzer is more precise and shows the ability to accurately classify normal versus abnormal sperm morphology. PMID- 17081527 TI - The rate at which serum total beta-subunit human chorionic gonadotropin increases after embryo transfer is a predictor of the viability of pregnancy and an identifier of determinants of pregnancy. AB - OBJECTIVE: To determine whether elements of treatment associated with faster doubling times of total beta-hCG in serum (beta-t2) in pregnant patients are also associated with a higher likelihood of pregnancy in all patients. DESIGN: Retrospective analysis of beta-t2 values, elements of ovarian stimulation (COH), and outcomes. SETTING: Private assisted reproductive technology (ART) center. PATIENT(S): Initial analysis of data from 432 cycles in which conception occurred after COH and embryo transfer, followed by analysis of pregnancy outcomes after 1,287 cycles of COH/ embryo transfer. INTERVENTION(S): No interventions. MAIN OUTCOME MEASURES: The beta-t2 values initially computed from consecutive serum beta-hCG levels in ongoing pregnancies were correlated with multiple properties of the patients and their treatment cycles. RESULT(S): The beta-t2 values during early pregnancy increased exponentially from about 1.6 days at 12 days to about 3.0 days at 24 days after embryo transfer. In those pregnancies which spontaneously aborted, early average beta-t2 values were higher than those for ongoing pregnancies; absolute beta-hCG levels did not differ. Positive correlations were established between beta-t2 values, the number of days of stimulation, and the number of ampules of drug administered per oocyte retrieved. The beta-t2 values were inversely related to average numbers of blastomeres in transferred embryos. Ongoing pregnancy rates (PR) were higher for cycles with lower gonadotropin dosages per oocyte retrieved, and when the average number of blastomeres in transferred embryos was higher. CONCLUSION(S): Steeper beta-hCG doubling times in early pregnancy were associated with lower gonadotropin dosages during ovarian stimulation and with higher numbers of blastomeres in transferred embryos. The latter variables were, in turn, associated with a higher likelihood of pregnancy after embryo transfer. PMID- 17081528 TI - Mifepristone-induced abortion in one horn and a growing fetus in another horn in a patient with a twin pregnancy in a bicornuate uterus. AB - OBJECTIVE: To report a case of mifepristone-induced abortion in only one horn but a growing fetus in another horn for a twin pregnancy in a bicornuate uterus. DESIGN: Case report. SETTING: Department of Obstetrics and Gynecology, Chang Gung Memorial Hospital, Taiwan. PATIENT(S): A 31-year-old woman who conceived spontaneously presented with 37 days' gestation. INTERVENTION(S): Mifepristone 600 mg orally followed by misoprostol 400 mug orally 2 days later. MAIN OUTCOME MEASURE(S): Termination of pregnancy. RESULT(S): Induced abortion in only one horn but a growing fetus in another horn for a twin pregnancy in the bicornuate uterus detected by ultrasound. Surgical abortion with vacuum curettage was performed, and menstruation resumed 1 month later. CONCLUSION(S): As medical abortion with mifepristone and misoprostol becomes more prevalent, more studies are necessary to establish the dosage and regimens for multiple pregnancies associated with uterine anomalies. PMID- 17081529 TI - Asian ethnicity is associated with reduced pregnancy outcomes after assisted reproductive technology. AB - OBJECTIVE: To determine whether success rates were similar in Asian and Caucasian women undergoing infertility treatment. DESIGN: Secondary data analysis and multivariate modeling. SETTING: Clinics reporting to the national Society for Assisted Reproductive Technology registry and a university-based clinic. PATIENT(S): Caucasian and self-identified Asian infertile women undergoing IVF. The study included 25,843 Caucasian and 1,429 Asian patients from the national registry; 370 Caucasian and 197 Asian patients were included from the site specific clinic. INTERVENTION(S): In vitro fertilization. MAIN OUTCOME MEASURE(S): Pregnancy rate and live-birth rate. RESULT(S): Infertile Asian women differed only minimally from their Caucasian counterparts in baseline characteristics and treatment response. Yet Asian women had a decreased clinical pregnancy rate (odds ratio, 0.71; 95% confidence interval 0.64-0.80) and a decreased live-birth rate (odds ratio, 0.69; 95% confidence interval 0.61-0.77). Subsequent multivariate analysis demonstrated that Asian ethnicity was an independent predictor of poor outcome. CONCLUSION(S): After treatment, infertile Asian women have significantly fewer pregnancies than do Caucasian women. Multivariate analysis indicates that this discrepancy cannot be accounted for by differences in baseline characteristics or by response to current therapeutic interventions. PMID- 17081530 TI - Sphingosine 1-phosphate protects ovaries from chemotherapy-induced damage in vivo. AB - OBJECTIVE: To investigate whether sphingosine-1-phosphate (S1P), an apoptosis inhibitor, would be able to protect ovarian follicles from chemotherapy-induced cell death in vivo. DESIGN: Animal study. SETTING: Academic medical center. ANIMAL(S): Twenty female mice. INTERVENTION(S): Twenty mice were randomly assigned into three groups: in group A (n = 8), each mouse received an injection of low concentration of S1P (A1: 0.5 mM), prepared in a vehicle (PET) into the bursa of one ovary and a high concentration (A2: 2.0 mM S1P) to the contralateral ovary. In group B (n = 8), only PET was injected to both ovaries. Afterwards, both groups received 100 mug Dacarbazine (Medac, Hamburg, Germany) IV. The control group C (n = 4) received no chemotherapy. After 2 weeks, the ovaries from group C and from 4 mice from group A and B were evaluated histologically. The remaining mice from group A and B were allowed 3 mating attempts at 4, 8, and 12 weeks after chemotherapy. MAIN OUTCOME MEASURE(S): Primordial/primary and pre /antral follicular density, pregnancy rates. RESULTS: Chemotherapy caused a significant reduction in the mean number of primordial follicles of mice treated with only PET or with low concentration of S1P (1.86/field of view [C] vs. 1.17 [B] and 0.98 [A1]; P=.006 and P<.0001, respectively) but not in the ovaries treated with high concentration of S1P (2.05/field of view [A2]; P=.918, not significant). Furthermore, three mice (75%) from group A became pregnant at the first mating attempt, in contrast to group B, in which only one mouse (25%) became pregnant and only after the third mating attempt. CONCLUSION(S): Local application of S1P protects ovarian follicles from chemotherapy-induced cell death, thereby preserving fertility. PMID- 17081531 TI - Antimullerian hormone predicts ovarian responsiveness, but not embryo quality or pregnancy, after in vitro fertilization or intracyoplasmic sperm injection. AB - Antimullerian hormone (AMH) and other markers of ovarian reserve were assessed to determine their predictive value with respect to treatment outcome. In a multivariate regression analysis, AMH was found to be predictive of the number of oocytes and the number of embryos, but not of embryo quality or the chance of a pregnancy, after IVF/ICSI. PMID- 17081532 TI - Dyspareunia and quality of sex life after laparoscopic excision of endometriosis and postoperative administration of triptorelin. AB - This observational cohort study examined the effect of laparoscopic full excision of endometriosis combined with postoperative triptorelin treatment on deep dyspareunia (DD) and quality of sex life. One year after completing the postoperative treatment, 45.9% of the patients had no DD and 34.7% reported a decrease in DD intensity; an increase in the frequency of sexual intercourse was reported by 62.2% of the women; objective improvements in several aspects of sex life were observed. PMID- 17081533 TI - Aberrant expression of deoxyribonucleic acid methyltransferases DNMT1, DNMT3A, and DNMT3B in women with endometriosis. AB - OBJECTIVE: Since endometriosis is a persistent disease with substantial gene dysregulation, there must be cellular memory of some sort that constitutes a unique cell identity for endometriotic cells. Epigenetic regulation, especially through DNA methylation, is a flexible, yet stable, mechanism for maintaining such a cellular memory. The aim of this study was to determine gene expression levels of DNMT1, DNMT3A, and DNMT3B, the three genes coding for DNA methyltransferases that are responsible for methylation. DESIGN: Cross-sectional measurements of gene expression levels of DNMT1, DNMT3A, and DNMT3B on endometriotic tissue. SETTING: Academic. PATIENT(S): Seventeen patients with laparoscopically confirmed endometriosis and 8 healthy women who underwent tubal sterilization who were free of endometriosis were recruited for the study. INTERVENTION(S): Epithelial cells were harvested from tissue samples by laser capture microdissection and messenger RNA abundance was measured by quantitative real-time reverse transcription-polymerase chain reaction. MAIN OUTCOME MEASURE(S): The expression levels of these genes in epithelial cells from 13 ectopic endometrial tissue samples, 10 eutopic endometrial tissue samples taken from women with endometriosis, and 8 normal endometrial tissue samples from women without endometriosis. RESULT(S): The genes DNMT1, DNMT3A, and DNMT3B were over expressed in the ectopic endometrium as compared with normal control subjects or the eutopic endometrium of women with endometriosis, and their expression levels were correlated positively with each other. CONCLUSION(S): The aberrant expression of these genes suggests that aberrant methylation may be rampant in endometriosis. This also provides a strong piece of evidence that endometriosis ultimately may be an epigenetic disease. PMID- 17081534 TI - Assessment of United States fertility clinic websites according to the American Society for Reproductive Medicine (ASRM)/Society for Assisted Reproductive Technology (SART) guidelines. AB - OBJECTIVE: To evaluate SART-member fertility clinic websites for their compliance with the 2004 ASRM/SART guidelines for advertising (which is deemed mandatory for clinic membership), to survey the general characteristics of the websites, and to assess differences between academic and private clinic websites. DESIGN: Cross sectional evaluation. SETTING: The Internet. PATIENTS: None. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Eleven objective criteria based on 2004 ASRM/SART guidelines for advertising and eight objective criteria for general characteristics of fertility clinic websites. RESULTS: All 384 SART-registered clinics were evaluated; 289 (75.3%) had functional websites (211 private, 78 academic). Success rates were published on 51% of websites (117 private, 31 academic), the majority of which were private clinics (p=.025). The percentage of fertility clinic websites adhering to ASRM/SART guidelines was low in all categories (ranging from 2.8%-54.5% in private centers and 1.3%-37.2% in academic centers). No statistically significant difference was found in the services offered at private versus academic clinics. CONCLUSION: A significant proportion of SART-member fertility clinics, both private and academic, that have websites are not following the ASRM/SART guidelines for advertising. Increased dissemination and awareness of the guidelines is warranted. PMID- 17081535 TI - Comparison of clomiphene citrate, metformin, or the combination of both for first line ovulation induction and achievement of pregnancy in 154 women with polycystic ovary syndrome. AB - OBJECTIVE: To determine which first-line medication is more effective in polycystic ovary syndrome (PCOS) patients for ovulation induction and pregnancy achievement and to verify whether any patient characteristic is associated with a better response to therapy. DESIGN: Observational comparative study. SETTING: Fertility clinic. PATIENT(S): One hundred fifty-four infertile women with oligomenorrhea and hyperandrogenism. INTERVENTION(S): Group 1 (56 patients) received clomiphene citrate (CC) 50 mg from days 5-9 of the cycle. Group 2 (57 patients) received 500 mg of metformin 3 times a day. Group 3 (41 patients) received both medications. MAIN OUTCOME MEASURE(S): Ovulation and pregnancy. RESULT(S): Patients receiving metformin alone had an increased ovulation rate compared with those receiving CC alone (75.4% vs. 50%). Patients on metformin had similar ovulation rates compared with those in the combination group (75.4% vs. 63.4%). Pregnancy rates were equivalent in the 3 groups. Response to metformin was independent of body weight and dose. Finally, nonsmoking predicted better ovulatory response overall as well as lower fasting glucose for CC and lower androgens for metformin. CONCLUSION(S): Metformin is better for ovulation induction than CC alone and equivalent for pregnancy achievement. We suggest that metformin can be used first for ovulation induction in patients with PCOS regardless of their weight and insulin levels because of its efficacy and known safety profile. PMID- 17081536 TI - Comparative study of the efficacy and tolerability of two vaginal progesterone formulations, Crinone 8% gel and Utrogestan capsules, used for luteal support. AB - OBJECTIVE: To compare the efficacy and tolerability of two different types of vaginal progesterone (P), Crinone 8% gel (Fleet Laboratories Ltd., Watford, United Kingdom) and Utrogestan capsules (Laboratories Besins International, Paris, France), used for luteal support after in vitro fertilization (IVF) cycles. DESIGN: Cohort study. SETTING: In Vitro Fertilization Polyclinic, Zagreb, Croatia. PATIENTS: A total of 285 women aged < or =37 years undergoing IVF-embryo transfer treatment. INTERVENTIONS: Patients were treated with either Crinone 8% vaginal P gel (90 mg) administered daily, or Utrogestan vaginal capsules (2 x 100 mg) administered three times daily. Progesterone was administered from the day of oocyte retrieval (day 0) to menses or, in a case of pregnancy, until week 12. MAIN OUTCOME MEASURE: Clinical pregnancy rate. The tolerability and acceptability of both preparations were determined by a questionnaire given to patients. RESULTS: The similar rates of clinical pregnancies (33.1% vs. 30.9%) [corrected] were obtained by using either Crinone 8% vaginal P gel or Utrogestan vaginal capsules. Overall tolerability and acceptability were significantly better in the Crinone group than in the Utrogestan group. CONCLUSIONS: The efficacy of the two vaginal P formulations was nearly the same, but the tolerability and acceptability of Crinone 8% gel were superior, in the opinion of patients. PMID- 17081537 TI - Relationship between endometrial thickness and embryo implantation, based on 1,294 cycles of in vitro fertilization with transfer of two blastocyst-stage embryos. AB - OBJECTIVE: To evaluate the relationship between endometrial thickness and clinical outcome of IVF and ET. DESIGN: Retrospective study. SETTING: Private assisted reproductive technology center. PATIENTS: One thousand two hundred and ninety-four infertility patients. INTERVENTIONS: IVF and fresh autologous ET of two blastocyst-stage embryos, including at least one good-quality blastocyst. MAIN OUTCOME MEASURES: Clinical pregnancy rate (PR) and spontaneous abortion rate. RESULTS: Endometrial thickness was greater in cycles resulting in pregnancy than in cycles not resulting in pregnancy (11.9 vs. 11.3 mm, respectively). Clinical pregnancy rates increased gradually from 53% among patients with a lining of <9 mm, to 77% among patients with a lining of > or =16 mm. Multiple logistic regression analysis indicated significant effects of age, embryo quality, and endometrial thickness on both clinical pregnancy rates and live birth or ongoing pregnancy rates. There was also a marginally significant trend toward decreasing rates of spontaneous pregnancy loss with increasing endometrial thickness. CONCLUSIONS: Clinical pregnancy and live-birth or ongoing pregnancy rates increase significantly with increasing endometrial thickness, independent of the effects of patient age and embryo quality. PMID- 17081538 TI - Selective cyclo-oxygenase-2 inhibition induces regression of autologous endometrial grafts by down-regulation of vascular endothelial growth factor mediated angiogenesis and stimulation of caspase-3-dependent apoptosis. AB - OBJECTIVE: To investigate the effects of selective cyclo-oxygenase-2 (COX-2) inhibition on the angiogenesis and proliferation of endometrial grafts. DESIGN: Intravital fluorescence microscopic study. SETTING: Institute for Clinical and Experimental Surgery, University of Saarland, Homburg/Saar, Germany. ANIMALS: Syrian golden hamsters. INTERVENTIONS: Endometrial fragments were transplanted into dorsal skinfold chambers of Syrian golden hamsters. Animals were treated daily with the selective COX-2-inhibitor NS398; controls received the vehicle dimethyl sulfoxide only. MAIN OUTCOME MEASURES: Angiogenesis was analyzed for 2 weeks with the use of intravital fluorescence microscopy. Protein expression of vascular endothelial growth factor, proliferating cell nuclear antigen, caspase 3, and activated caspase-3 was measured by Western blot analysis. Histological sections were scanned for local microthrombosis. RESULTS: COX-2 inhibition induced a marked regression of endometrial grafts due to inhibition of angiogenesis, as indicated by significantly reduced microvessel density within grafts compared to controls. This effect was associated with a decreased expression of vascular endothelial growth factor. Moreover, COX-2 inhibition suppressed cell proliferation and induced apoptosis-associated caspase-3 expression. Interestingly, microthrombus formation could not be observed. CONCLUSIONS: Our study demonstrates that selective COX-2 inhibition induces regression of endometrial grafts by suppression of angiogenesis and stimulation of apoptosis. Accordingly, COX-2 inhibition may represent a novel therapeutic strategy for the treatment of endometriosis. PMID- 17081539 TI - The histone to protamine ratio in human spermatozoa: comparative study of whole and processed semen. AB - Sperm protamine deficiency has been demonstrated in some infertile men and may be an important determinant of sperm DNA integrity. We sought to examine and compare the sperm nuclear histone to protamine ratio in whole and processed semen samples from fertile and infertile men. PMID- 17081540 TI - Efficacy of intravenous tramadol treatment for reducing pain during office diagnostic hysteroscopy. AB - OBJECTIVE: To assess whether IV tramadol before outpatient hysteroscopy could reduce procedure-related pain. DESIGN: A randomized double-blind placebo controlled trial. SETTING: Outpatient Hysteroscopy Centre in the Department of Obstetrics and Gynaecology of Cagliari University. PATIENT(S): Fifty healthy, parous, women who underwent outpatient diagnostic hysteroscopy and endometrial biopsy. INTERVENTION(S): Random IV infusion of tramadol or placebo before hysteroscopy and endometrial biopsy were performed. MAIN OUTCOME MEASURE(S): Visual analogue scale of pain was measured both immediately after and 15 minutes after the procedure. Stress hormones (ACTH, cortisol), blood pressure, and heart frequency were evaluated before, during, and 15 minutes after the procedure. RESULT(S): In the tramadol group, the visual analogue scale of pain was significantly lower than in the placebo group both immediately after the procedure and 15 minutes later. Basal levels of ACTH and cortisol did not differ between the groups. In both groups, the ACTH levels remained unchanged during the study, and the cortisol levels were higher 15 minutes after the procedure than before the procedure. Procedure time, heart frequency, blood pressure, and adverse effects did not differ between the groups. CONCLUSION(S): In parous women without uterine malformations, a treatment with tramadol before hysteroscopy and endometrial biopsy appears to be capable of reducing the pain and discomfort that are associated with this procedure. PMID- 17081541 TI - Social dominance regulates androgen and estrogen receptor gene expression. AB - In Astatotilapia burtoni, dominant males have higher levels of sex steroid hormones than subordinate males. Because of the complex regulatory interactions between steroid hormones and receptors, we asked whether dominance is also associated with variation in sex steroid receptor gene expression. Using quantitative PCR, we compared the expression of specific subtypes of androgen (AR) and estrogen (ER) receptor genes between dominant and subordinated males in 3 divisions of the brain, the pituitary, and the testes. We measured mRNA levels of AR-alpha, AR-beta, ER-alpha, ER-betaa, and ER-betab, gonadotropin-releasing hormone 1 (GnRH1), and GnRH receptor 1 (GnRH-R1) relative to 18S rRNA. In the anterior part of the brain, we found that dominant males had higher mRNA expression of AR-alpha, AR-beta, ER-betaa, and ER-betab, but not ER-alpha, compared to subordinate males. This effect of dominance was reflected in a positive correlation between testes size and AR-alpha, AR-beta, ER-betaa, and ER betab in the anterior brain. In addition, mRNA levels of all ARs and ERs in the anterior brain were positively correlated with mRNA level of GnRH1. In the middle and posterior portions of the brain, as well as the testes, steroid receptor mRNA levels were similar among dominants and subordinates. In the pituitary, ER-alpha mRNA level was positively correlated with testes size and AR-alpha mRNA was positively correlated with GnRH-R1 mRNA level. These data suggest that dominant male brains could be more sensitive to sex steroids, which may contribute to the increased complexity of the behavioral repertoires of dominant males. PMID- 17081542 TI - The impact of human immunodeficiency virus (HIV) on outcome and practice in trauma: past, present and future. AB - Since the initial description of a concentrated outbreak of pneumocystis carnii pneumonia in 1981, the human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) pandemic has accounted for nearly 25 million deaths worldwide. This review focuses on estimations of prevalence by geographic region and identification of high-risk populations within each region, outcome for trauma patients with HIV and AIDS and risk management for health care workers who sustain occupational exposures. Trauma surgeons are more likely to encounter patients infected with HIV in geographic areas where HIV prevalence is high or in areas where intravenous drug use, high-risk sexual behaviours and penetrating trauma are more common. Patients with HIV may be expected to have higher rates of infectious and respiratory complications if they have active AIDS and/or liver disease caused by one of the hepatitis viruses. Certain aspects of therapy may change in this group of patients. Clinicians should be aware that highly active anti-retroviral therapy (HAART) might produce complications. Occupational exposure among healthcare workers is uncommon. Cases of infection in healthcare workers from needlesticks are rare. Certain precautions regarding body fluid and needlestick exposures have been widely adopted over the past decade. When percutaneous injury results in known exposure to HIV, post-exposure prophylaxis (PEP) should be used and can be expected to be effective in preventing infection in the large majority of cases. PMID- 17081543 TI - Dyspnoea and chest pain associated with pulmonary polymethylmethacrylate embolism after percutaneous vertebroplasty. PMID- 17081544 TI - Re: surgical management of the severely displaced supracondylar fracture of the humerus in children. PMID- 17081545 TI - Outcome after surgical decompression of severe traumatic brain injury. AB - One of the factors that affects outcome following severe traumatic brain injury is development and progression of cerebral oedema with associated increase in intracranial pressure (ICP). Uncontrolled elevations of ICP may compromise energy metabolism of the injured brain and lead to secondary injury, affecting neurological outcome of the patient. Decompressive craniectomy has been used for over a century as a treatment of refractory brain swelling in a variety of neurological conditions. However, conclusive evidence of whether it has a beneficial or adverse affect on outcome is lacking. This article reviews the existing evidence on the role of decompressive craniectomy in management of patients with traumatic brain injury and stresses the need for randomised controlled trials. PMID- 17081546 TI - The flaming gypsy skirt injury. AB - On review of admissions over a 12-month period, we noted a significant number of women presenting with gypsy skirt burns. We describe all six cases to highlight the unique distribution of the wounds and the circumstances in which the accidents occurred. Four skirts were ignited by open fire heaters: two skirts ignited whilst the women were standing nearby, distracted with a telephone conversation; one brushed over the flame as she was walking past the heater; other whilst dancing in the lounge. One skirt was ignited by decorative candles placed on the floor during a social gathering. Another skirt was set alight by cigarette ember, whilst smoking in the toilet. Percentage surface area burned, estimated according to the rule of nines, showed that gypsy skirt burns were significant ranging from 7 to 14% total body surface area (TBSA) and averaging 9% TBSA. Two patients required allogenic split-skin grafts. Common sense care with proximity to naked flame is all that is needed to prevent this injury. PMID- 17081547 TI - Assessing cross-cultural validity of scales: a methodological review and illustrative example. AB - In this article, we assessed the cross-cultural validity of the Women's Role Strain Inventory (WRSI), a multi-item instrument that assesses the degree of strain experienced by women who juggle the roles of working professional, student, wife and mother. Cross-cultural validity is evinced by demonstrating the measurement invariance of the WRSI. Measurement invariance is the extent to which items of multi-item scales function in the same way across different samples of respondents. We assessed measurement invariance by comparing a sample of working women in Taiwan with a similar sample from the United States. Structural equation models (SEMs) were employed to determine the invariance of the WRSI and to estimate the unique validity variance of its items. This article also provides nurse-researchers with the necessary underlying measurement theory and illustrates how SEMs may be applied to assess cross-cultural validity of instruments used in nursing research. Overall performance of the WRSI was acceptable but our analysis showed that some items did not display invariance properties across samples. Item analysis is presented and recommendations for improving the instrument are discussed. PMID- 17081548 TI - In vitro study of foot kinematics using a dynamic walking cadaver model. AB - There is a dearth of information on navicular, cuboid, cuneiform and metatarsal kinematics during walking and our objective was to study the kinematic contributions these bones might make to foot function. A dynamic cadaver model of walking was used to apply forces to cadaver feet and mobilise them in a manner similar to in vivo. Kinematic data were recorded from 13 cadaver feet. Given limitations to the simulation, the data describe what the cadaver feet were capable of in response to the forces applied, rather than exactly how they performed in vivo. The talonavicular joint was more mobile than the calcaneocuboid joint. The range of motion between cuneiforms and navicular was similar to that between talus and navicular. Metatarsals four and five were more mobile relative to the cuboid than metatarsals one, two and three relative to the cuneiforms. This work has confirmed the complexity of rear, mid and forefoot kinematics. The data demonstrate the potential for often-ignored foot joints to contribute significantly to the overall kinematic function of the foot. Previous emphasis on the ankle and sub talar joints as the principal articulating components of the foot has neglected more distal articulations. The results also demonstrate the extent to which the rigid segment assumptions of previous foot kinematics research have over simplified the foot. PMID- 17081549 TI - Separation and identification of isomeric acidic degradation products of organophosphorus chemical warfare agents by capillary electrophoresis-ion trap mass spectrometry. AB - Capillary electrophoresis (CE) coupled to ion trap mass spectrometry (MS) was evaluated for the separation and identification of chemical warfare agent degradation products (alkylphosphonic acids and alkyl alkylphosphonic acids). Different analytical parameters were optimized in negative ionization mode such as electrolyte composition (15 mM CH(3)COONH(4), pH 8.8), sheath liquid composition (MeOH/H(2)O/NH(3), 75:25:2, v/v/v), nebulization and ion trapping conditions. A standard mixture of five alkylphosphonic (di)acids and five alkyl alkylphosphonic (mono)acids containing isomeric compounds was used in order to evaluate CE selectivity and MS identification capability. The obtained electropherograms revealed that CE selectivity was very limited in the case of alkyl alkylphosphonic acid positional isomers, whereas isomeric isopropylphosphonic and propylphosphonic acids were baseline-separated. CE-MS-MS experiments provided an unambiguous identification of each isomeric co-migrating alkyl alkylphosphonic acids thanks to the presence of specific fragment ions. On the other hand, CE separation was mandatory for the identification of isomeric alkylphosphonic acids, which led to the same fragment ion and could not be differentiated by MS-MS. The developed method was applied to the analysis of soil extracts spiked with the analytes (before or after extraction treatment) and appeared to be very promising since resolution and sensitivity were similar to those observed in deionized water. Especially, analytes were detected and identified in soil extract spiked at 5 microg mL(-1) with each compound before extraction treatment. PMID- 17081550 TI - Comparison of the behavior of 13C- and deuterium-labeled polycyclic aromatic hydrocarbons in analyses by isotope dilution mass spectrometry in combination with pressurized liquid extraction. AB - We examined the differences found in the concentrations of polycyclic aromatic hydrocarbons (PAHs) in sediment samples when determined by isotope dilution mass spectrometry (IDMS) using 13C-labeled PAHs (13C-PAHs) and deuterium-labeled PAHs (PAHs-d) as internal standards. The samples were analyzed by pressurized liquid extraction (PLE) in combination with gas chromatography-mass spectrometry. The observed concentrations with PAHs-d as internal standards were significantly (p<0.05) lower (1.9-4.3%) than those determined with 13C-PAHs. The most important factor responsible for the high recovery of PAHs-d relative to 13C-PAHs and native PAHs is the higher stability of PAHs-d during the PLE process. PMID- 17081551 TI - Chromatographic behaviour of peptides on a mixed-mode stationary phase with an embedded charged group by capillary electrochromatography and high-performance liquid chromatography. AB - Retention behaviour of biological peptides was investigated on a stationary phase bearing an embedded quaternary ammonium group in a C21 alkyl chain by both high performance liquid chromatography (HPLC) and capillary electrochromatography (CEC). In HPLC experiments, variation of acetonitrile (ACN) content in the mobile phase showed that peptides are mainly separated by RP mechanism. The weak or negative retention factors observed as compared to C18 silica stationary phase suggested the involvement of an electrostatic repulsion phenomenon in acidic conditions. Comparison of HPLC and CEC studies indicated that (i) ion-exclusion phenomenon is more pronounced in HPLC and (ii) higher ACN percentage in mobile phase induce for some peptides an increase of retention in CEC, pointing out the existence of mechanisms of retention other than partitioning mainly involved in chromatographic process. This comparative study demonstrated the critical role of electric field on peptide retention in CEC and supports the solvatation model of hydrolytic pillow proposed by Szumski and Buszewski for CEC using mixed mode stationary phase in CEC. PMID- 17081552 TI - Validation of methodology for simultaneous determination of synthetic dyes in alcoholic beverages by capillary electrophoresis. AB - In this work a method of analysis for synthetic dyes was developed using capillary electrophoresis in alcoholic beverages. The analyses were carried out with fused silica capillary, with 73 cm effective length, at 35 degrees C, buffer phosphate solution of 10 mmol/L with sodium dodecyl sulphate 10 mmol/L, pH 11, and +25 kV of voltage. For dye analyses, three wavelengths in the visible region were used for the qualitative and quantitative determination of the 11 synthetic dyes allowed in Brazil: 450, 525 and 625 nm for the yellow, red and blue dyes, respectively. The detection limits varied from 0.4 to 2.5 microg/mL and the quantification limits varied from 1.3 to 7.1 microg/mL. The average recovery was 92.6 and 104.0% at two levels of concentration. Repeatability for standards and spiked sample showed that the calculated values were greater than the observed values, demonstrating the precision of the method. The proposed and validated method was used to analyze some alcoholic beverage samples, consisting of 12 red wines, 9 coolers, 6 aromatized spirits, 7 bitters, 3 cocktails and 8 liquors from different Brazilian manufacturers. The results showed the coolers, bitters and red wines did not have synthetic dyes, but dyes were found in six of the eight analyzed liquor samples. In all the samples of cocktails and spirits, the presences of dyes were observed. No analyzed sample exceeded the limit established by Brazilian legislation (maximum 30 mg/100 mL). PMID- 17081553 TI - A water-activated pump for portable microfluidic applications. AB - An on-chip micropump for portable microfluidic applications was investigated using mathematical modeling and experimental testing. This micropump is activated by the addition of water, via a dropper, to ionic polymer particles that swell due to osmotic effects when wetted. The resulting particle volume increase deflects a membrane, forcing a separate fluid from an adjacent reservoir. The micropump components, along with the microfluidic components, are fabricated using the contact liquid photolithographic polymerization (CLiPP) method. The maximum flow rate achieved with this pump is 17 microL per minute per mg of dry polymer particles of 355-425 microm in diameter. The pump flow rate may be controlled by adjusting the particle size and amount, the membrane properties, and the channel dimensions. The experimental results demonstrate good agreement with an analytical model describing the particle swelling and its coupling with resistive forces from the bending membrane, viscous flow in the microchannel, and interfacial effects. Key features of this micropump are that it can be placed directly on a microdevice, and that it requires only a small amount of water and no external power supply to function. Therefore, this pumping system is useful for applications in which a highly portable device is required. PMID- 17081554 TI - Surface energy of talc and chlorite: Comparison between electronegativity calculation and immersion results. AB - The surface energies of talc and chlorite is computed using a simple model, which uses the calculation of the electrostatic energy of the crystal. It is necessary to calculate the atomic charges. We have chosen to follow Henry's model of determination of partial charges using scales of electronegativity and hardness. The results are in correct agreement with a determination of the surface energy obtained from an analysis of the heat of immersion data. Both results indicate that the surface energy of talc is lower than the surface energy of chlorite, in agreement with observed behavior of wettability. The influence of Al and Fe on this phenomenon is discussed. Surface energy of this type of solids seems to depend more strongly on the geometry of the crystal than on the type of atoms pointing out of the surface; i.e., the surface energy depends more on the physics of the system than on its chemistry. PMID- 17081555 TI - Enhanced aqueous solubilization of tetrachloroethylene by a rhamnolipid biosurfactant. AB - A rhamnolipid biosurfactant produced by Pseudomonas aeruginosa ATCC 9027 was isolated, purified and characterized in terms of its ability to mobilize and solubilize tetrachloroethylene (PCE) for potential use in surfactant-enhanced aquifer remediation (SEAR) applications. Using a drop volume method, the PCE biosurfactant steady-state interfacial tension was determined and found to be ca. 10 mN/m which is not low enough to cause significant PCE nonaqueous phase liquid (NAPL) mobilization. It was observed that the biosurfactant partitioned significantly into PCE at aqueous concentrations higher than the critical micelle concentration (CMC). After accounting for rhamnolipid partitioning into the PCE phase, a weight solubilization ratio (WSR) of 1.2 g(PCE)/g(rhamnolipid) was determined and through this mechanism the biosurfactant significantly improved the apparent aqueous solubility of PCE. PMID- 17081556 TI - The early repolarization variant--normal or a marker of heart disease in certain subjects. AB - The theory is advanced that the early repolarization variant may not always be benign and that it can become a substrate for ventricular arrhythmias, sudden death, and hypercontractility cardiomyopathy in some subjects, including certain high-performance athletes. In addition, it is suggested that it likely represents part of a spectrum of cardiovascular anomalies related to nonischemic ST elevation including Brugada syndrome, and that it may also have a molecular genetic origin of variable penetrance. PMID- 17081557 TI - Standardized analysis for the quantification of Vbeta CDR3 T-cell receptor diversity. AB - Assessment of the diversity of the T-cell receptor (TCR) repertoire is often determined by measuring the frequency and distribution of individually rearranged TCRs in a population of T cells. Spectratyping is a common method used to measure TCR repertoire diversity, which examines genetic variation in the third complementarity-determining region (CDR3) region of the TCR Vbeta chain using RT PCR length-distribution analysis. A variety of methods are currently used to analyze spectratype data including subjective visual measures, qualitative counting measures, and semi-quantitative measures that compare the original data to a standard, control data set. Two major limitations exist for most of these approaches: data files become very wieldy and difficult to manage, and current analytic methods generate data which are difficult to compare between laboratories and across different platforms. Here, we introduce a highly efficient method of analysis that is based upon a normal theoretical Gaussian distribution observed in cord blood and recent thymic emigrants. Using this analysis method, we demonstrate that PBMC obtained from patients with various diseases have skewed TCR repertoire profiles. Upon in vitro activation with anti CD3 and anti-CD28 coated beads (Xcyte Dynabeads) TCR diversity was restored. Moreover, changes in the TCR repertoire were dynamic in vivo. We demonstrate that use of this streamlined method of analysis in concert with a flexible software package makes quantitative assessment of TCR repertoire diversity straightforward and reproducible, enabling reliable comparisons of diversity values between laboratories and over-time to further collaborative efforts. Analysis of TCR repertoire by such an approach may be valuable in the clinical setting, both for prognostic potential and measuring clinical responses to therapy. PMID- 17081558 TI - Induction and termination of prepupal summer diapause in Pseudopidorus fasciata (Lepidoptera: Zygaenidae). AB - The seasonal life cycle of the zygaenid moth, Pseudopidorus fasciata is complicated by two different developmental arrests: a winter diapause as a fourth larval instar and a summer diapause as a prepupa in a cocoon. Both larval diapause induction and termination are under photoperiodic control. Short days induce larval diapause with a critical daylength of 13.5h and long days terminate diapause with a critical daylength of 14h. In the present study photoperiodic control of summer diapause was investigated in Pseudopidorus fasciata. Under long photoperiods ranging from LD 14:10 to LD 18:6, only part of the population entered summer diapause, the rest continued to develop. The lowest number of prepupae entered diapause at LD 14:10, followed by LD 16:8 and LD 17:7. The highest incidence of diapause occurred with photoperiods of LD 15:9 and LD 18:6. By transferring the diapausing prepupae induced by various long photoperiods (LD 14:10, LD 15:9, LD 16:8, LD 17:7, LD 18:6) to LD 13:11, 25 degrees C, the duration of diapause induced by LD 14:10 was significantly shorter than those induced by longer photoperiods. By keeping aestivating prepupae induced by LD 15:9, 28 degrees C or by natural conditions at short photoperiods (LD 11:13 and LD 13:11) and at a long photoperiod (LD 15:9), the duration of diapause at LD 15:9 was more than twice as long as than those at LD 11:13 and LD 13:11. Moreover, adult emergence was highly dispersed with a high mortality at LD 15:9 but was synchronized with low mortality at LD 11:13 and LD 13:11. When the naturally induced aestivating prepupae were kept under natural conditions, the early aestivating prepupae formed in May exhibited a long duration of diapause (mean 126 days), whereas the later-aestivating prepupae formed in July exhibited a short duration of diapause (mean 69 days). These results indicate that aestivating prepupae require short or shortening photoperiod to terminate their diapause successfully. By transferring naturally induced aestivating prepupae to 25, 28 and 30 degrees C, the duration of diapause at the high temperature of 30 degrees C was significantly longer than those at 25 and 28 degrees C, suggesting that high temperature during summer also plays an important role in the maintenance of summer diapause in Pseudopidorus fasciata. All results reveal that summer diapause can serve as a "bet hedging" against unpredictable risks due to fluctuating environments or as a feedback mechanism to synchronize the period of autumn emergence. PMID- 17081559 TI - Effects of an unidentified microsporidium on the convergent lady beetle, Hippodamia convergens Guerin-Meneville (Coleoptera: Coccinellidae), used for biological control. AB - Convergent lady beetles, Hippodamia convergens Guerin-Meneville, are collected from overwintering sites in California and redistributed for aphid control in home gardens and agroecosystems. The effects of an unidentified microsporidium on the life history characteristics of commercially available H. convergens were examined. Mean development for microsporidia-infected and uninfected H. convergens was 15.40+/-0.14 and 14.76+/-0.16 days, respectively (P=0.01). Larval mortality did not differ significantly. Cumulative mean egg production for microsporidia-infected and uninfected females was 545.8+/-92.6 and 928.3+/-86.4 eggs, respectively (P=0.004) and mean survival was 64.5+/-5.6 and 77.1+/-4.5 days, respectively (P=0.04). Microsporidian spores (3.6x2.4 microm) are similar in size to those of Nosema hippodamiae. PMID- 17081560 TI - Induction of antioxidant and detoxification response by oxidants in cardiomyocytes: evidence from gene expression profiling and activation of Nrf2 transcription factor. AB - Mild or low doses of oxidants are known to prime cells towards resistance against further damage. In cardiomyocytes, we found that pretreatment with 100 microM H(2)O(2) prevents the cells from apoptosis induced by doxorubicin (Dox). Affymetrix microarray analyses of 28,000 genes reveal that H(2)O(2) treated cells reduced expression of genes encoding cytochrome c, mitochondrial complex I, III, IV and V and several contractile proteins. Elevated expression of antioxidant and detoxification genes appears as a dominant feature of the gene expression profile of H(2)O(2) treated cells. Most of the genes in this category contain an Antioxidant Response Element (ARE) in their promoters. Measurements of ARE promoter-reporter gene activity indicate a dose- and time-dependent activation of the ARE by H(2)O(2). Since the Nrf2 transcription factor regulates ARE-mediated gene expression, we overexpressed Nrf2 to test whether activation of Nrf2 is sufficient to induce cytoprotection. High levels of Nrf2 expression were achieved via adenovirus mediated gene delivery. Transduced Nrf2 was present in the nuclei and caused an increase in the expression of NAD(P)H:quinone oxidoreductase 1 (NQO1), a representative downstream target of Nrf2. Unlike H(2)O(2) pretreated cells, the cells expressing high levels of Nrf2 were not resistant to Dox-induced apoptosis. Therefore, the cytoprotective effect of H(2)O(2) pretreatment is not reliant upon Nrf2 activation alone as measured by resistance against Dox-induced apoptosis. PMID- 17081561 TI - Troponin phosphorylation and regulatory function in human heart muscle: dephosphorylation of Ser23/24 on troponin I could account for the contractile defect in end-stage heart failure. AB - We made quantitative measurements of phosphorylation in troponin isolated from 6 non-failing donor hearts and 6 explanted hearts with end-stage heart failure in SDS-PAGE gels using Pro-Q Diamond phosphoprotein stain. The troponin T phosphorylation level was the same in troponin from failing and non-failing heart (3.1 mol Pi/mol). However, troponin I phosphorylation was significantly lower in failing (0.37+/-0.18 mol Pi/mol) compared with non-failing heart troponin (2.25+/ 0.36 mol Pi/mol). Levels of troponin I PKA-dependent phosphorylation, measured with a phosphoserine 23/24-specific antibody, were also significantly lower in failing heart troponin (0.19+/-0.06 mol Pi/mol) compared to non-failing troponin (1.14+/-0.09 mol Pi/mol). We calculate that there is phosphorylation in addition to serine 23/24 of 1.11+/-0.34 mol Pi/mol in non-failing reduced to 0.18+/-0.17 mol Pi/mol in failing heart troponin, attributed to phosphorylation on the PKC sites. To test for the functional role of troponin I phosphorylation, the native troponin I from either non-failing or failing heart troponin was exchanged for a recombinant (unphosphorylated) human cardiac troponin I. Thin filament Ca(2+) regulatory function was studied with the quantitative in vitro motility assay: thin filaments containing the replaced troponin I resulted in a failing phenotype of a 17-26% reduced sliding speed and an increased Ca(2+)-sensitivity relative to non-failing troponin (EC(50) TnI-exchanged/non-failing=0.57, p<0.001). When exchanged with troponin I phosphorylated with PKA motility parameters reverted to a pattern indistinguishable from non-failing troponin (p=0.35-0.75). We suggest that changes in troponin function can account for the contractile abnormality in failing heart muscle and that the functional changes in troponin are due to reduced phosphorylation of troponin I at the PKA sites. PMID- 17081562 TI - Ryanodine receptors and ventricular arrhythmias: emerging trends in mutations, mechanisms and therapies. AB - It has been six years since the first reported link between mutations in the cardiac ryanodine receptor Ca(2+) release channel (RyR2) and catecholaminergic polymorphic ventricular tachycardia (CPVT), a malignant stress-induced arrhythmia. In this time, rapid advances have been made in identifying new mutations, and in understanding how these mutations disrupt normal channel function to cause VT that frequently degenerates into ventricular fibrillation (VF) and sudden death. Functional characterisation of these RyR2 Ca(2+) channelopathies suggests that mutations alter the ability of RyR2 to sense its intracellular environment, and that channel modulation via covalent modification, Ca(2+)- and Mg(2+)-dependent regulation and structural feedback mechanisms are catastrophically disturbed. This review reconciles the current status of RyR2 mutation-linked etiopathology, the significance of mutational clustering within the RyR2 polypeptide and the mechanisms underlying channel dysfunction. We will also review new data that explores the link between abnormal Ca(2+) release and the resultant cardiac electrical instability in VT and VF, and how these recent developments impact on novel anti-arrhythmic therapies. Finally, we evaluate the concept that mechanistic differences between CPVT and other arrhythmogenic disorders may preclude a common therapeutic strategy to normalise RyR2 function in cardiac disease. PMID- 17081563 TI - Structural model of human endoglin, a transmembrane receptor responsible for hereditary hemorrhagic telangiectasia. AB - Endoglin is a type I membrane protein expressed as a disulphide-linked homodimer on human vascular endothelial cells whose haploinsufficiency is responsible for the dominant vascular dysplasia known as hereditary hemorrhagic telangiectasia (HHT). Structurally, endoglin belongs to the zona pellucida (ZP) family of proteins that share a ZP domain of approximately 260 amino acid residues at their extracellular region. Endoglin is a component of the TGF-beta receptor complex, interacts with the TGF-beta signalling receptors types I and II, and modulates cellular responses to TGF-beta. Here, we have determined for the first time the three-dimensional structure of the approximately 140 kDa extracellular domain of endoglin at 25 A resolution, using single-particle electron microscopy (EM). This reconstruction provides the general architecture of endoglin, which arranges as a dome made of antiparallel oriented monomers enclosing a cavity at one end. A high resolution structure of endoglin has also been modelled de novo and found to be consistent with the experimental reconstruction. Each subunit comprises three well-defined domains, two of them corresponding to ZP regions, organised into an open U-shaped monomer. This domain arrangement was found to closely resemble the overall structure derived experimentally and the three modelled de novo domains were tentatively assigned to the domains observed in the EM reconstruction. This molecular model was further tested by tagging endoglin's C terminus with an IgG Fc fragment visible after 3D reconstruction of the labelled protein. Combined, these data provide the structural framework to interpret endoglin's functional domains and mutations found in HHT patients. PMID- 17081565 TI - Mechanism of formation of actomyosin interface. AB - Force generation in muscle results from binding of myosin to F-actin. ATP binding to myosin provides energy to dissociate actomyosin complex while the hydrolysis of ATP is needed for re-binding of myosin to F-actin. At the end of each cycle myosin and actin form a tight complex with a substantial interface area. We investigated the dynamics of formation of actomyosin interface in presence and absence of nucleotides by quenched flow cross-linking technique. We showed previously that myosin head (subfragment 1, S1) directly interacts with at least two monomers in the actin filament. The quenched flow cross-linking experiments revealed that the initial contact (in presence or absence of nucleotides) occurs between loop 635-647 of S1 and 1-12 N-terminal residues of one actin and, then, the second contact forms between loop 567-574 of S1 and the N terminus of the second actin. The distance between these two loops in S1 corresponds to the distance between N termini of two actins in the same strand (53 A) but is smaller than that between two actins from the different strands (102 A). The formation of the actomyosin complex proceeds in ordered sequence: S1 initially binds to one actin then binds with the second actin located in the same strand but probably closer to the barbed end of F-actin. The presence of nucleotides slows down the interaction of S1 with the second actin, which correlates with recently proposed cleft movement in a 50 kDa domain of S1. The sequential mechanism of formation of actomyosin interface starting from one end and developing towards the barbed end might be involved in force generation and directional movement in actin-myosin system. PMID- 17081564 TI - Involvement of DEAD-box proteins in group I and group II intron splicing. Biochemical characterization of Mss116p, ATP hydrolysis-dependent and independent mechanisms, and general RNA chaperone activity. AB - The RNA-catalyzed splicing of group I and group II introns is facilitated by proteins that stabilize the active RNA structure or act as RNA chaperones to disrupt stable inactive structures that are kinetic traps in RNA folding. In Neurospora crassa and Saccharomyces cerevisiae, the latter function is fulfilled by specific DEAD-box proteins, denoted CYT-19 and Mss116p, respectively. Previous studies showed that purified CYT-19 stimulates the in vitro splicing of structurally diverse group I and group II introns, and uses the energy of ATP binding or hydrolysis to resolve kinetic traps. Here, we purified Mss116p and show that it has RNA-dependent ATPase activity, unwinds RNA duplexes in a non polar fashion, and promotes ATP-independent strand-annealing. Further, we show that Mss116p binds RNA non-specifically and promotes in vitro splicing of both group I and group II intron RNAs, as well as RNA cleavage by the aI5gamma-derived D135 ribozyme. However, Mss116p also has ATP hydrolysis-independent effects on some of these reactions, which are not shared by CYT-19 and may reflect differences in its RNA-binding properties. We also show that a non-mitochondrial DEAD-box protein, yeast Ded1p, can function almost as efficiently as CYT-19 and Mss116p in splicing the yeast aI5gamma group II intron and less efficiently in splicing the bI1 group II intron. Together, our results show that Mss116p, like CYT-19, can act broadly as an RNA chaperone to stimulate the splicing of diverse group I and group II introns, and that Ded1p also has an RNA chaperone activity that can be assayed by its effect on splicing mitochondrial introns. Nevertheless, these DEAD-box protein RNA chaperones are not completely interchangeable and appear to function in somewhat different ways, using biochemical activities that have likely been tuned by coevolution to function optimally on specific RNA substrates. PMID- 17081567 TI - Apoptosis and functional changes of dipeptide transporter (PepT1) in the rat small intestine after traumatic brain injury. AB - BACKGROUND: Traumatic brain injury (TBI) can induce significant alterations of intestinal mucosal structure and barrier function. However, it has not been investigated whether, and to what degree, apoptosis and alterations of absorptive function in the intestinal mucosal epithelium occur after TBI. MATERIAL AND METHODS: Male Wistar rats were randomly divided into seven groups (five rats each group) including normal group, control group with sham operation, and TBI groups at hours 3, 12, 24, and 72, and on day 7. Parietal brain contusion was adopted using weight-dropping method. Intestinal mucosal structure was examined using histomorphmetric study and electron microscopy, and apoptosis was detected by TUNEL method. An everted sleeve of intestine was securely incubated in Kreb's solution with radioactive dipeptide ((3)H-Gly-Sar, 10 microCi/mL) to measure the uptake and transport of PepT1 of small intestinal epithelial cells. RESULTS: The villous height, crypt depth and surface area were significantly decreased at 24 h after TBI, and further declined to the degree of mucosal atrophy on day 7 after TBI. Apoptotic changes of condensed nuclei in epithelial cells and fractured, distorted, and sparse microvilli were found by electron microscopy. The number of apoptotic cells in the mucosal epithelium was significantly increased since 3 h after TBI, peaked at 72 h post-injury, then declined at 7 days, but was still higher than that of control. There was a highly negative relation between the apoptotic index and the villous height, the crypt depth, and villous surface area. Compared with that of normal and control rats, the transport and uptake of dipeptide was significantly increased at 3 h post-injury (P < 0.01), peaked at 12 h and declined a bit at 24 h post-injury, and returned to the level of normal and control rats at 72 h and 7 days. CONCLUSIONS: It is highly suggested that intestinal mucosa apoptosis plays an important role in the pathogenesis of acute gut damage after TBI. Intestinal PepT1 expression could be up-regulated after traumatic brain injury, and maintained the normal level under the condition of serious intestinal damage. Up-regulation of PepT1 may adaptively improve absorption of di- and tripeptides, independent of changes in the mucosal surface area. PMID- 17081566 TI - A bioartificial liver device secreting interleukin-1 receptor antagonist for the treatment of hepatic failure in rats. AB - BACKGROUND: Liver transplantation is the treatment of choice for many patients with fulminant hepatic failure (FHF). A major limitation of this treatment is the lack of available donors. An optimally functioning bio-artificial liver (BAL) device has the potential to provide critical hepatic support to patients with FHF. In this study, we examined the efficacy of combining interleukin-1 (IL-1) receptor blockade with the synthetic function of hepatocytes in a BAL device for the treatment of FHF. MATERIALS AND METHODS: We injected an adenoviral vector encoding human IL-1 receptor antagonist (AdIL-1Ra) into the liver of D galactosamine (GalN) intoxicated rats via the portal vein. We also transfected primary rat hepatocytes and reversibly immortalized human hepatocytes (TTNT cells) with AdIL-1Ra, and incorporated these transfected hepatocytes into our flat-plate BAL device and evaluated their efficacy in our GalN-induced FHF rat model after 10 h of extracorporeal perfusion. RESULTS: Rats injected with AdIL 1Ra showed significant reductions in the plasma levels of hepatic enzymes. Primary rat hepatocytes transfected with AdIL-1Ra secreted IL-1Ra without losing their original synthetic function. Incorporating these cells into the BAL device and testing in a GalN-induced FHF rat model resulted in significant reductions in plasma IL-6 levels and significantly improved animal survival. Incorporating the AdIL-1Ra transfected TTNT cells in the BAL device and testing in the GalN-induced FHF rat model resulted in significantly reduced plasma IL-6 levels, and a trend toward improved survival was seen. CONCLUSION: Hepatocytes producing IL-1Ra are a promising cell source for BAL devices in the treatment of GalN-induced FHF. PMID- 17081568 TI - Liver regeneration and hemodynamics in pigs with mesocaval shunt. AB - BACKGROUND: A surgical technique using a mesocaval shunt and downstream ligation of the superior mesenteric vein has been recently proposed to overcome the size limitations that restrict the use of partial liver grafts. We designed an experimental study in pigs to evaluate the capacities of liver regeneration and hemodynamic changes after completion of this procedure. MATERIAL AND METHODS: Liver regeneration after left hepatectomy was compared between two groups of five pigs, with or without mesocaval shunt, sacrificed 11 to 14 days after surgery. A third group of five animals was used for hemodynamic studies. RESULTS: Liver regeneration in study animals was 45.3% of controls. This was obtained despite a reduction of the venous inflow to 15.6% of the control, resulting in a net decrease of the total blood inflow to 56% of the control, despite a compensatory increase in the arterial inflow. There was no significant difference in mitotic index, hepatocellular size, and glycogen content between study and control animals. CONCLUSION: Our experimental study confirms that the regenerative capacities of the pig liver are largely preserved despite the dramatic reduction of the venous blood inflow, reduced to its gastroduodenosplenopancreatic component. This lends further support to the hypothesis that the gastroduodenosplenopancreatic blood is enriched in hepatotrophic factors, likely to originate from the pancreas and duodenum. PMID- 17081569 TI - Kinetic behaviour of proenzymes activation in the presence of different inhibitors for both activating and activated enzymes. AB - In the present paper, a kinetic analysis of a general model for proenzyme activation, where the activating enzyme and also the activated one are reversibly inhibited in two steps by two different inhibitors, has been performed. The cases in which both inhibitors are the same, or in which the inhibition is irreversible (only one or the two inhibition routes) are treated as particular cases of the general model. In addition, the kinetic behaviour of many other proenzyme activation systems involving inhibition, particular cases of the reaction scheme under study, can be obtained. The total number of particular cases for the general model under study is 370, so this approach offers to the scientific community working in limited proteolysis regulation for the first time a method based on general solutions which only needs to be specified to their concrete problem of zymogen activation. Finally, new adimensional parameters are introduced, allowing the knowledgement, in the case that any of the inhibition routes is irreversible, the relative weight of both activation and irreversible inhibition routes. PMID- 17081570 TI - A quantitative observation and imaging of single tumor cell migration and deformation using a multi-gap microfluidic device representing the blood vessel. AB - A microfluidic device was developed for quantifying the migratory and deformability capabilities of a single tumor cell using direct imaging. It was fabricated using photolithography and is made of polydimethysiloxane. Chemotaxis approach was used for directing cell movement, using 10 microm microgaps to restrict the migration to a single cell. Each cell's migration rate is quantified as a measure of its distance traveled over time taken. Real-time recording of cell deformation under physiological flow was performed, and the elongation index and surface area change of the cells were compared. Three human tumor cell lines viz. HepG2, HeLa and MDA-MB-435S were used to verify the operation and methodology of the device. Their migration rates ranged from 5 to 15 microm/h, consistent with other scientific reports. By reducing the microgap width to 3 microm, it was found that the cells moved along the row of microgaps but were unable to migrate across the microgaps. Subsequent deformation of the cells through the gaps further showed that their migratory capability might be governed by their deformation ability and the deformation stress on their membranes. The strategy of targeting cancer cell membrane for rupture may provide a therapy for metastasis. Being a valuable tool for rapid quantification of a single cell's migratory capability, this device should be helpful for pharmacologic and drug screening, investigation of factors that regulate cell migration and deformation. PMID- 17081571 TI - Chronic food restriction: enhancing effects on drug reward and striatal cell signaling. AB - Chronic food restriction (FR) increases behavioral sensitivity to drugs of abuse in animal models and is associated with binge eating, which shares comorbidity with drug abuse, in clinical populations. Behavioral, biochemical and molecular studies conducted in this laboratory to elucidate the functional and mechanistic bases of these phenomena are briefly reviewed. Results obtained to date indicate that FR increases the reward magnitude and locomotor-activating effects of abused drugs, and direct dopamine (DA) receptor agonists, as a result of neuroadaptations rather than changes in drug disposition. Changes in striatal DA dynamics, and postsynaptic cell signaling and gene expression in response to D-1 DA receptor stimulation have been observed. Of particular interest is an upregulation of NMDA receptor-dependent MAP kinase and CaM Kinase II signaling, CREB phosphorylation, and immediate-early and neuropeptide gene expression in nucleus accumbens (NAc) which may facilitate reward-related learning, but also play a role in the genesis of maladaptive goal-directed behaviors. Covariation of altered drug reward sensitivity with body weight loss and recovery suggests a triggering role for one of the endocrine adiposity hormones. However, neither acute nor chronic central infusions of leptin or the melanocortin 3/4 receptor agonist, MTII, have attenuated d-amphetamine reward or locomotor activation in FR rats. Interestingly, chronic intracerebroventricular leptin infusion in ad libitum fed (AL) rats produced a sustained decrease in food intake and body weight that was accompanied by a reversible potentiation of rewarding and locomotor-activating effects of d-amphetamine. This raises the interesting possibility that rapid progressive weight loss is sufficient to increase behavioral sensitivity to drugs of abuse. Whether weight loss produced by leptin infusion produces the same neuroadaptations as experimenter-imposed FR, and whether any of the observed neuroadaptations are necessary for expression of increased behavioral responsiveness to acute drug challenge remain to be investigated. PMID- 17081572 TI - Altered sleep-wake cycles and food intake: the Ramadan model. AB - In this review the effects of diurnal fasting on normal physiological processes are considered. Ramadan is placed in a circadian context, food and fluid ingestion being displaced to the pre-sunrise and post-sunset hours. Over the holy month, negative energy balance is often experienced, though this deficit is not a universal finding. Responses to exercise during the day show influences consistent with hypohydration and an increased reliance on fat as a source of fuel for exercise. Muscle performance and psychomotor performance are impaired as the month of fasting progresses but it is not clear how circadian rhythms in responses to activity are altered. For some measures at rest there is a reduction in amplitude and a delay in acrophase. Health-related benefits are reflected in a rise in high-density lipoprotein cholesterol and individuals with predispositions for coronary heart disease are not at increased risk of cardiovascular disorders due to fasting. The physiological adjustments during the month have some similarities to the disturbances in circadian rhythms experienced in different circumstances. The Ramadan model provides an alternative to those for ageing, nocturnal shift-work and time-zone transitions in understanding the links between behaviour and endogenous circadian rhythms. PMID- 17081573 TI - Effect of an afternoon confectionery snack on cognitive processes critical to learning. AB - Two experiments examined how an afternoon confectionery snack affects a variety of cognitive processes critical to learning. For Experiment 1, thirty-eight male undergraduates completed a dual learning task where the primary task involved learning either a map or stories and the secondary task required monitoring a radio broadcast for a specific word category. Results showed that for map learning, participants who consumed the confectionery snack performed better on the primary task. They correctly placed more country names and left fewer blanks on a map during long-term recall. However, on the secondary attention task, participants who consumed the confectionery snack had a lower hit rate. The confectionary snack did not affect story memory performance. In Experiment 2, 38 boys, aged 9-11 years, participated in a similar, age appropriate task. Results showed that boys who had consumed the confectionery snack correctly placed more names and left fewer blanks on a map in both short-term and long-term recall. In contrast with Experiment 1, performance on the secondary task was better after confectionary consumption. However, when tested on a separate vigilance attention task, children who consumed the placebo performed better. Overall results indicate that a confectionery snack, ingested in the afternoon, generally improves spatial memory, but has a mixed effect on attention performance. PMID- 17081574 TI - Consumption of a mid-morning snack improves memory but not attention in school children. AB - Muthayya, S., T. Thomas, K. Srinivasan, K. Rao, A. V. Kurpad, J.-W. Van Klinken, G. Owen and E.A. de Bruin: Consumption of a mid-morning snack improves memory but not attention in school children. Physiol Behav 00(0) 000-000, 2006.--This study aimed to determine whether consumption of a mid-morning snack with appropriate energy compensation through a smaller breakfast or lunch, resulted in improved cognitive performance of 7-9 year old children with a low and high socioeconomic status (LSES and HSES, n=35 and 34 respectively). The children were each randomly assigned to three iso-caloric dietary interventions: control (standard breakfast, no snack and standard lunch), intervention A (small breakfast, snack, and standard lunch) and intervention B (standard breakfast, snack, and small lunch), using a cross-over design. The children were tested on three different days, each one week apart. Computerised tests of cognitive performance, consisting of memory, sustained attention and psychomotor speed, were performed during four sessions, i.e., prior to breakfast, after breakfast, after a mid-morning snack and after lunch. Having a mid-morning snack resulted in a smaller decline in immediate and delayed memory in LSES but not in HSES children. Having a snack did not influence sustained attention and psychomotor speed in either LSES or HSES children. This study shows that a more evenly distributed energy intake throughout the morning by consuming a mid-morning snack improves memory performance in school-age LSES children even when the total amount of energy consumed during the morning is not altered. PMID- 17081575 TI - Identification and expression regulation of symbiotically activated legume genes. AB - Legume plants are able to enter two different endosymbioses with soil prokaryotes and soil fungi, leading to nitrogen-fixing root nodules and to arbuscular mycorrhiza (AM), respectively. We applied in silico and microarray-based transcriptome profiling approaches to uncover the transcriptome of developing root nodules and AM roots of the model legume Medicago truncatula. Several hundred genes were found to be activated in different stages of either symbiosis, with almost 100 genes being co-induced during nodulation and in arbuscular mycorrhiza. These co-induced genes can be associated with different cellular functions required for symbiotic efficiency, such as the facilitation of transport processes across the perisymbiotic membranes that surround the endosymbiotic bacteroids in root nodules and the arbuscules in AM roots. To specify promoter elements required for gene expression in arbuscule-containing cells, reporter gene fusions of the promoter of the Vicia faba leghemoglobin gene VfLb29 were studied by loss-of-function and gain-of-function approaches in transgenic hairy roots. These analyses specified a 85-bp fragment that was necessary for gene expression in arbuscule-containing cells but was dispensible for gene activation in root nodules. In contrast to promoters mediating gene expression in the infected cells of root nodules, the activation of genes in AM appears to be governed by more complex regulatory systems requiring different promoter modules. PMID- 17081576 TI - Development of bioinformatic tools to support EST-sequencing, in silico- and microarray-based transcriptome profiling in mycorrhizal symbioses. AB - The great majority of terrestrial plants enters a beneficial arbuscular mycorrhiza (AM) or ectomycorrhiza (ECM) symbiosis with soil fungi. In the SPP 1084 "MolMyk: Molecular Basics of Mycorrhizal Symbioses", high-throughput EST sequencing was performed to obtain snapshots of the plant and fungal transcriptome in mycorrhizal roots and in extraradical hyphae. To focus activities, the interactions between Medicago truncatula and Glomus intraradices as well as Populus tremula and Amanita muscaria were selected as models for AM and ECM symbioses, respectively. Together, almost, 20.000 expressed sequence tags (ESTs) were generated from different random and suppressive subtractive hybridization (SSH) cDNA libraries, providing a comprehensive overview of the mycorrhizal transcriptome. To automatically cluster and annotate EST-sequences, the BioMake and SAMS software tools were developed. In connection with the eNorthern software SteN, plant genes with a predicted mycorrhiza-induced expression were identified. To support experimental transcriptome profiling, macro- and microarray tools have been constructed for the two model mycorrhizae, based either on PCR-amplified cDNAs or 70mer oligonucleotides. These arrays were used to profile the transcriptome of AM and ECM roots under different conditions, and the data obtained were uploaded to the ArrayLIMS and EMMA databases that are designed to store and evaluate expression profiles from DNA arrays. Together, the EST- and transcriptome databases can be mined to identify candidate genes for targeted functional studies. PMID- 17081577 TI - Triterpenoid saponins and phenylethanoid glycosides from stem of Akebia trifoliata var. australis. AB - A detailed phytochemical study on the 70% aqueous ethanol extract of stems of Akebia trifoliata (Thunb.) Koidz. var. australis (Diels) Rehd led to isolation of five compounds, together with 12 known triterpenoid saponins and three known phenylethanoid glycosides. The structures of the five compounds were elucidated on the basis of analysis of spectroscopic data and physicochemical properties as: 2alpha, 3beta, 23-trihydroxy-30-norolean-12-en-28-oic acid beta-D-glucopyranosyl ester (1), 2alpha, 3beta, 23-trihydroxy-30-norolean-12-en-28-oic acid beta-D xylopyranosyl-(1-->3)-O-alpha-D-rhamnopyranosyl-(1-->4)-O-beta-D-glucopyranosyl (1-->6)-O-beta-D-glucopyranosyl ester (2), 2alpha, 3beta, 23-trihydroxyurs-12-en 28-oic acid beta-D-xylopyranosyl-(1-->3)-O-alpha-L-rhamnopyranosyl-(1-->4)-O-beta D-glucopyranosyl-(1-->6)-O-beta-D-glucopyranosyl ester (3), 3-beta-[(beta-D glucopyranosyl-(1-->3)-O-alpha-L-arabinopyranosyl)oxy]-23-hydroxy-30-norolean-12 en-28-oic acid alpha-L-rhamnopyranosyl-(1-->4)-O-beta-D-glucopyranosyl-(1-->6)-O beta-D-glucopyranosyl ester (4) and 3-beta-[(alpha-L-xylopyranosyl-(1-->2)-O alpha-L-arabinopyranosyl)oxy]-30-norolean-12-en-28-oic acid alpha-L rhamnopyranosyl-(1-->4)-O-beta-D-glucopyranosyl-(1-->6)-O-beta-D-glucopyranosyl ester (5), named mutongsaponin A, B, C, D and E, respectively. PMID- 17081578 TI - Gene expression analysis of arbuscule development and functioning. AB - The arbuscular mycorrhiza (AM) is characterized by specific morphological structures of the fungus and the plant and by physiological adaptations which are mostly beneficial for both partners of the symbiosis. This review describes approaches to study the molecular basis of the interaction. RNA accumulation patterns have been monitored in Pisum sativum to analyse the plant response to arbuscule development. In a direct approach, the Mtha1 gene from Medicago truncatula was cloned which is expressed in arbusculated cells of M. truncatula. The gene putatively encodes an H(+)-ATPase involved in the improved plant nutrition during mycorrhization. Finally, a tripartite system between M. truncatula, Glomus mosseae and Aphanomyces euteiches was established, in order to study bioprotection. Analysis of the transcriptome has been started to analyse the interaction between the plant, the pathogen and the AM fungus. PMID- 17081579 TI - Proceedings of the 14th UKPHA annual forum, 2006. PMID- 17081580 TI - Novel paradigms on scorpion toxins that affects the activating mechanism of sodium channels. AB - Scorpion toxins classified as beta-class are reviewed using a new paradigm. Four distinct sub types are recognized: "classical", "Tsgamma-like", "excitatory" and "depressant"beta-scorpion toxins. Recent experimental data have made possible to identify the interacting interfaces of the Na(+) channel-receptor site 4 with some of these toxins. The voltage-sensor trapping mechanism proposed for the action of these toxic peptides is analyzed in the context of what causes a modification of the activating mechanism of Na(+) channels. A cartoon model is presented with the purpose of summarizing the most current knowledge on the field. Finally, the recent advances on the knowledge of the specific interactions of beta-toxins and different sub types of Na(+) channels are also reviewed. PMID- 17081581 TI - Characterization of the Epstein-Barr virus glycoprotein BMRF-2. AB - Epstein-Barr virus (EBV) BMRF-2 protein interaction with the beta1 family of integrins plays an important role in EBV infection of polarized oral epithelial cells. In this work, we characterized BMRF-2 protein expression in EBV-infected B lymphoblastoid and polarized oral epithelial cells, and in hairy leukoplakia (HL) epithelium. BMRF-2 expression in B cells and polarized oral epithelial cells was associated with the EBV lytic infection. In these cells, BMRF-2 is efficiently transported to the cell membrane and its integrin binding Arg-Gly-Asp (RGD) motif is exposed on the cell surface. BMRF-2 is highly expressed in HL epithelium and accumulates at the lateral border of oral keratinocytes. In EBV-infected polarized oral epithelial cells, this protein is transported to the basolateral membranes and co-localized with beta1 integrin. These data suggest that BMRF-2 may play an important role in cell-to-cell spread of EBV within the oral epithelium. BMRF-2 is glycosylated through O-linked oligosaccharides; it forms oligomers and is associated with the virion envelope. Its C-terminal tail is localized in the cytoplasm. We found that beta1, alpha5, and alpha3 integrins are present in purified EBV virions. We show that BMRF-2 is a ligand for beta1, alpha5, alpha3, and alphav integrins and our data are consistent with a role for BMRF-2 in viral lytic infection. PMID- 17081582 TI - Mapping antigenic diversity and strain specificity of mumps virus: a bioinformatics approach. AB - Mumps is an acute infectious disease caused by mumps virus, a member of the family Paramyxoviridae. With the implementation of vaccination programs, mumps infection is under control. However, due to resurgence of mumps epidemics, there is a renewed interest in understanding the antigenic diversity of mumps virus. Hemagglutinin-neuraminidase (HN) is the major surface antigen and is known to elicit neutralizing antibodies. Mutational analysis of HN of wild-type and vaccine strains revealed that the hypervariable positions are distributed over the entire length with no detectable pattern. In the absence of experimentally derived 3D structure data, the structure of HN protein of mumps virus was predicted using homology modeling. Mutations mapped on the predicted structures were found to cluster on one of the surfaces. A predicted conformational epitope encompasses experimentally characterized epitopes suggesting that it is a major site for neutralization. These analyses provide rationale for strain specificity, antigenic diversity and varying efficacy of mumps vaccines. PMID- 17081583 TI - Peripheral fading with monocular and binocular viewing. AB - This study measured the fading times of peripheral targets as a function of whether viewing was monocular or binocular, and of brightness contrast. Data from a binocularly normal group showed Troxler fading to be significantly faster with monocular (i.e., patched) than with binocular viewing. In contrast, one-eyed observers showed significantly longer fading times than the two-eyed observers viewing monocularly and equivalent times to their binocular viewing. A control experiment showed that these findings were not due to worse fixation stability, larger pupil sizes, or an unusually large blinking rate in the enucleated group. The enucleated group actually exhibited a slight miosis, equivalent fixation stability, and a normal blinking rate. In both experiments, the times to fading of all observers were a function of brightness contrast. We conclude that in binocularly normal observers patching or closing one eye does not produce monocular vision but rather a condition of weak binocular rivalry, and that the absence of inhibitory binocular interactions in the enucleated group may explain, in part, their resistance to fading and their superior performance in other contrast-defined tasks. PMID- 17081584 TI - Influence of history on saccade countermanding performance in humans and macaque monkeys. AB - The stop-signal or countermanding task probes the ability to control action by requiring subjects to withhold a planned movement in response to an infrequent stop signal which they do with variable success depending on the delay of the stop signal. We investigated whether performance of humans and macaque monkeys in a saccade countermanding task was influenced by stimulus and performance history. In spite of idiosyncrasies across subjects several trends were evident in both humans and monkeys. Response time decreased after successive trials with no stop signal. Response time increased after successive trials with a stop signal. However, post-error slowing was not observed. Increased response time was observed mainly or only after cancelled (signal inhibit) trials and not after noncancelled (signal respond) trials. These global trends were based on rapid adjustments of response time in response to momentary fluctuations in the fraction of stop signal trials. The effects of trial sequence on the probability of responding were weaker and more idiosyncratic across subjects when stop signal fraction was fixed. However, both response time and probability of responding were influenced strongly by variations in the fraction of stop signal trials. These results indicate that the race model of countermanding performance requires extension to account for these sequential dependencies and provide a basis for physiological studies of executive control of countermanding saccade performance. PMID- 17081586 TI - Implications of a kinematic wave model for first flush treatment design. AB - A deterministic model was developed to predict pollutant mass first flush and to utilize it for better design of best management practices (BMPs) that focus on treating the first flush. The model used the kinematic wave equation to calculate flow and mass transport, and erosion equations to calculate pollutant concentrations, which were assumed to be from a short and a long term source. The model parameters were calibrated with a parameter estimation procedure using three years' monitoring data from a highway runoff site in west Los Angeles. The simulation results showed that there exists an optimum watershed size to maximize first flush. Contours of watershed length, developed from simulations for different conditions of rainfall and watershed geometry, can be used to design runoff collection systems for highways and parking lots to maximize first flush. PMID- 17081585 TI - Target recovery in metacontrast: the effect of contrast. AB - The visibility of a target stimulus (T) can be reduced by an aftercoming and spatially non-overlapping mask stimulus (M1), a phenomenon known as metacontrast masking. Interestingly, the visibility of the masked target can be recovered when a secondary mask (M2) is added to the T-M1 sequence. We analyzed a computational model of retino-cortical dynamics (RECOD) and derived the prediction that contrast dependence of metacontrast and target recovery should parallel the contrast dependence of afferent magnocellular and parvocellular pathways, respectively. In a psychophysical experiment, we tested this prediction by systematically varying (a) M2's contrast and (b) the M1-M2 onset asynchrony (SOA). At the optimal M1-M2 SOA, target recovery effect increased with M2's contrast without saturating, but at the optimal M1-M2 metacontrast SOA, reduction of M1's visibility saturated very rapidly as M2's contrast increased. Quantitative comparisons of psychophysical results with model simulations provide support for our prediction. We conclude that metacontrast masking is driven by signals originating from the magnocellular pathway and target recovery in metacontrast is driven by signals originating from the parvocellular pathway. PMID- 17081587 TI - Source apportionment of PAHs in sediments using factor analysis by time records: application to Lake Michigan, USA. AB - Apportionment of sources of polycyclic aromatic hydrocarbons (PAHs) by a factor analysis model based on time records for seven sediment cores from central Lake Michigan is reconsidered to provide a more quantitative treatment, and one additional factor (source). The effect of mixing depth was observed on the unmixed fluxes, and a computer program was developed for estimation of the average PAH fluxes from the reconstructed PAH records. The new factor analysis model with nonnegative constraints includes not only scaling but also back scaling which improved the source apportionment significantly. One core, CLM-A, was an outlier and was omitted from the analysis. Wood burning was identified as a third major source of PAHs in the area (20% of the total PAH flux). The other two sources petroleum (traffic), 45%, and coal (coke oven emissions), 35%, became more apparent, where the contributions are based on the sum of concentrations of 16 PAH compounds. A reduced mixing depth (1.0-1.5cm) was found to be more appropriate than the originally estimated mixing depth (1.5-2.0cm). PMID- 17081588 TI - Formation of chlorinated estrones via hypochlorous disinfection of wastewater effluent containing estrone. AB - Chlorinated derivatives of estrone (E1) in the effluent of a municipal sewage treatment plant located in Shizuoka prefecture, Japan were detected by gas chromatography/mass spectrometry using electron impact in selected ion monitoring (GC/MS-EI-SIM) analysis. The concentrations of E1, 2-chloroestrone, 4 chloroestrone and 2,4-dichloroestrone in the effluent sample collected in December 2005 were 60.0 ng l(-1), 4.0 ng l(-1), 14.5 ng l(-1), and 9.8 ng l(-1), respectively. In the effluent sample taken in June 2005, 2,4-dichloroestrone was detected at 5.6 ng l(-1) along with 17.6 ng l(-1) of E1. However, only E1 was detected at 5.9 ng l(-1) in the sample in May 2005. To elucidate the behavior of E1 during the disinfection process with sodium hypochlorite in the sewage treatment plant, we carried out a reaction of E1 with sodium hypochlorite in buffer solutions at pH 7 and 9. As E1 was consumed rapidly, chlorinated estrones were produced and relatively fast formation of 2-chloroestrone, 4-chloroestrone, and 2,4-dichloroestrone was observed. Furthermore, 1,4-estradiene-3,17-dione derivatives were formed from the reaction between 2,4-dichloroestrone and sodium hypochlorite. PMID- 17081589 TI - Urinary arsenic methylation and porphyrin profile of C57Bl/6J mice chronically exposed to sodium arsenate. AB - Arsenic interferes with the function of enzymes responsible for haem biosynthesis leading to alteration in the porphyrin profile. In this study, young female C57Bl/6J mice were given drinking water containing 0, 100, 250 and 500 microg As(V)/L as sodium arsenate ad libitum for 24 months. 24 h pooled urine samples were collected bimonthly for urinary arsenic methylation and porphyrin analyses by HPLC-ICP-MS and HPLC respectively. The levels of total arsenic were significantly dose related except for the 2nd month interval. No significant differences in the urinary arsenic methylation pattern between control and test groups were observed. Coproporphyrin I (Copro I) showed a significant dose response relationship after 12, 14 and 20 months of exposure. Significant differences in the levels of coproporphyrin III (Copro III) were observed in the 8th month in 250 and 500 microg/L treatment groups and the dose-response pattern was maintained after 10 and 12 months. Our results suggest that urinary arsenic is a useful biomarker for internal dose, and that urinary coproporphyrin can be used as an early warning biomarker of effects before the onset of cancer. PMID- 17081590 TI - Changes of soil organic carbon in an intensively cultivated agricultural region: a denitrification-decomposition (DNDC) modelling approach. AB - Regional modelling of soil organic carbon (SOC) dynamics is important for predicting large-scale patterns in carbon cycling and for assessing potential responses of soil carbon pools to land-use change. However, data uncertainties related to both spatial heterogeneity and small-scale differences in farming practice related to cropping systems affect the accuracy of regional models. A case study is presented from Quzhou County in the North China Plain, an area characterized by highly intensive farming. For this county, the DNDC model was validated using sampling data from 68 sites around the county under generalized farm practices. Unique modelling units based on soil type, soil texture and crop type were created and then used to model the spatial change of SOC under different farming practices. Considering the main factors affecting SOC sequestration, the results indicate that the DNDC model delivers acceptable modelling results at county level. The results show that there is a great potential for SOC sequestration in Quzhou County in its central, southern and eastern parts. Changes in farming practices show a strong effect on carbon sequestration. A very efficient and environmental friendly sequestration of SOC pools could be achieved even by decreasing nitrogen fertilizer inputs, when the amount of straw returned to the field is greatly increased. PMID- 17081591 TI - Effects of soil compaction and irrigation on the concentrations of selenium and arsenic in wheat grains. AB - Arsenic is a known human carcinogen, whereas Se is essential to human health and human Se intake in some European countries has decreased in recent decades. There have been few reports on the potential effect of soil physical conditions on the uptake of Se and As by food crops. Field experiments were conducted to evaluate the effects of soil compaction and irrigation on Se and As concentrations in wheat grains in two seasons. Grain Se concentration varied from 10 to 115 microg kg(-1), whereas the concentration of As in grain was low (<20 microg kg(-1)). Irrigation significantly decreased grain Se concentration by 30-75%, and the effect was likely due to a dilution as a result of increased grain yield, a competition of sulfate added in the irrigation water on Se uptake and increased leaching losses. Soil compaction significantly decreased grain Se concentration in one season. In contrast, soil compaction increased grain As concentration significantly when the crop was irrigated. The contrasting effects of soil compaction on As and Se uptake are explained by the difference in their mobility in soil and the pathway of ion transport to roots. The observed effect on grain Se is significant for human and animal nutrition, because the concentration varied from sufficient to very low levels in response to the alternation of soil physical conditions in the field. Soil physical conditions have to be taken into account when assessing the bioavailability of the contaminant As or the essential nutrient Se. PMID- 17081592 TI - Dynamics of phosphorus, nitrogen and carbon removal in a horizontal subsurface flow constructed wetland. AB - The dynamics of nitrogen (N), phosphorus (P) and carbon (C) accumulation in the filter material of a horizontal subsurface constructed wetland (HSSF CW; established in 1997) and in a specially designed oil-shale ash filter (2002) for P retention have been studied. Concentrations of N, P and C in filter media (coarse sand) in the HSSF beds show an increasing trend. Both the annual accumulation of P and increasing outflow concentrations of P in the HSSF CW reflect the possible saturation of filter media with P after 8 years working. Tested ash material derived from oil-shale combustion demonstrated very high P removal efficiency in laboratory batch experiments. However, during the first 4 months of the in situ ash filter experiment, the efficiency of P removal was about 71% (an average outflow concentration of 1.9 mg L(-1) was achieved). Subsequently, the efficiency decreased to 10-20%, which might be a sign of saturation or clogging due to quick biofilm development on the ash particles. The increasing of hydraulic retention time and the improvement of design for maximal contact between material and wastewater are considered to be key factors that can provide optimal pH for the removal processes. PMID- 17081593 TI - Magnitude of arsenic pollution in the Mekong and Red River Deltas--Cambodia and Vietnam. AB - Large alluvial deltas of the Mekong River in southern Vietnam and Cambodia and the Red River in northern Vietnam have groundwaters that are exploited for drinking water by private tube-wells, which are of increasing demand since the mid-1990s. This paper presents an overview of groundwater arsenic pollution in the Mekong delta: arsenic concentrations ranged from 1-1610 microg/L in Cambodia (average 217 microg/L) and 1-845 microg/L in southern Vietnam (average 39 microg/L), respectively. It also evaluates the situation in Red River delta where groundwater arsenic concentrations vary from 1-3050 microg/L (average 159 microg/L). In addition to rural areas, the drinking water supply of the city of Hanoi has elevated arsenic concentrations. The sediments of 12-40 m deep cores from the Red River delta contain arsenic levels of 2-33 microg/g (average 7 microg/g, dry weight) and show a remarkable correlation with sediment-bound iron. In all three areas, the groundwater arsenic pollution seem to be of natural origin and caused by reductive dissolution of arsenic-bearing iron phases buried in aquifers. The population at risk of chronic arsenic poisoning is estimated to be 10 million in the Red River delta and 0.5-1 million in the Mekong delta. A subset of hair samples collected in Vietnam and Cambodia from residents drinking groundwater with arsenic levels >50 microg/L have a significantly higher arsenic content than control groups (<50 microg/L). Few cases of arsenic related health problems are recognized in the study areas compared to Bangladesh and West Bengal. This difference probably relates to arsenic contaminated tube-well water only being used substantially over the past 7 to 10 years in Vietnam and Cambodia. Because symptoms of chronic arsenic poisoning usually take more than 10 years to develop, the number of future arsenic related ailments in Cambodia and Vietnam is likely to increase. Early mitigation measures should be a high priority. PMID- 17081594 TI - Increase in carbon emissions from forest fires after intensive reforestation and forest management programs. AB - This paper shows an example of substantial increase in carbon emissions from forest fires after reforestation on a national scale. It is the first estimation of historical carbon emissions from forest fires in Korea during the last 40 years. Investigation was focused on the recent increase in large forest fires and its closely related factors. A simple modeling approach to estimate carbon emission was applied. The direct carbon emission from forest fires in 2000, ranging from 115 to 300 Gg C, corresponds to 1-3% of the annual carbon uptake by forests. The influence of forest fires on the carbon cycle in Korea is not so significant, but Korean forests have a large potential for generating severe local fires due to increasing forest carbon density and a high forest area ratio (forest area/total land area) of 65%. The carbon emission per area burned (Mg C ha(-1)) clearly reflects the trend toward increases in the number of severe fires. Statistical analyses and the trends of annual temperature and precipitation show that the recent large increase in carbon emissions may be the negative consequences of intensive forest regrowth that is the product of successful reforestation and forest management programs rather than the effect of climate change. These results imply a need for further studies in other countries, where large-scale plantation has been conducted, to evaluate the role of plantation and forest fires on the global carbon cycle. PMID- 17081595 TI - Lead fluxes, isotopic and concentration profiles in a peat deposit near a lead smelter (Pribram, Czech Republic). AB - The content and the isotopic composition of lead (Pb) were studied in a peat deposit on the ridge of the Brdy Hills, in the vicinity of the Pribram metallurgical works, in the Czech Republic. Quadrupole ICP MS was employed to determine the elemental composition and (206)Pb/(207)Pb and (208)Pb/(206)Pb isotope ratios. The individual layers were dated using alpha spectrometric measurement of the (210)Pb activity. The historical time period covered by the studied cores reached back to the 18th century. The Pb concentration in the studied profiles varied from 10 to 550 mg kg(-1). The (206)Pb/(207)Pb ratio varied in the range from 1.154 to 1.194 in the individual parts of the profile. The metallurgy of the Pb ores ((206)Pb/(207)Pb approximately 1.16), lithogenic Pb ((206)Pb/(207)Pb approximately 1.2), metallurgical processing of automobile batteries ((206)Pb/(207)Pb approximately 1.17) and the combustion of coal ((206)Pb/(207)Pb approximately 1.17-1.19) yield isotopic signatures that determine the isotope compositions of the individual profiles. Deposition rates between 15 mg m(-2) year(-1) at the beginning of the 19th century and 320 mg m( 2) year(-1) in the 1980s were determined in the dated profiles. The increased deposition rates determined on the dated profiles correspond to the increasing production of Pb ores in the Pribram mining area at the turn of the 19th and 20th centuries. The maximum for metallurgical production corresponds to the highest deposition rates recorded in 1960s and 1970s. The current deposition rate of 5-89 mg m(-2) year(-1) Pb is related to erosion of contaminated soils and waste dumps. PMID- 17081596 TI - Interactions of rice (Oryza sativa L.) and PAH-degrading bacteria (Acinetobacter sp.) on enhanced dissipation of spiked phenanthrene and pyrene in waterlogged soil. AB - The effects of cultivation of rice (Oryza sativa L.) and PAH-degrading bacteria (Acinetobacter sp.) separately, and in combination, on the dissipation of spiked phenanthrene and pyrene (0, 50+50, 100+100, 200+200 mg kg(-1)) in waterlogged soil were studied using pot trials. The population of introduced PAH-degrading bacteria remained at 10(5) CFU g(-1) dry soil after 20 days of treatment with Acinetobacter sp. only, but increased to 10(6) when planted with rice simultaneously. Shoot and root biomass of rice when grown alone was adversely affected by spiked PAHs, but significantly increased by 2-55% and 8-409%, respectively, when inoculated with Acinetobacter sp.. Phenanthrene and pyrene concentrations in roots ranged from 1-27 and 20-98 mg kg(-1), respectively, while their concentrations in shoots were generally lower than 0.2 mg kg(-1). The dissipation of phenanthrene was mainly due to abiotic loss as 70-78% phenanthrene was lost from the control soil at the end of 80 days, while removal of 86-87% phenanthrene had been achieved after 40 days in the treatment co-cultivated with Acinetobacter sp. and rice. Compared with the control where only 6-15% of pyrene was removed from soil, a much higher dissipation of pyrene (43-62%) was attained for the treatments co-cultivated with Acinetobacter sp. and rice at the end of 80 days. The results demonstrated that co-cultivation of rice and PAH-degrading bacteria may have a great potential to accelerate the bioremediation process of PAH-contaminated soil under waterlogged conditions. PMID- 17081597 TI - Metrology for ultrasonic applications. AB - This paper provides a review of current metrological capability applied to the characterisation of the acoustic output of equipment used within medical ultrasonic applications. Key measurement devices, developed to underpin metrology in this area, are the radiation force balance, used to determine total output power, and the piezo-electric hydrophone, used to resolve the spatial and temporal distribution of acoustic pressure. The measurement infrastructure in place within the United Kingdom ensuring users are able to carry out traceable measurements of these quantities in a meaningful way, is described. This includes the relevant primary standards, the way international equivalence of national standards is demonstrated and the routes by which the standards are disseminated to the user community. Emerging measurement techniques that may in future lead to improved measurement capability, are also briefly discussed. PMID- 17081598 TI - Modified HPV16 E7/HSP70 DNA vaccine with high safety and enhanced cellular immunity represses murine lung metastatic tumors with downregulated expression of MHC class I molecules. AB - OBJECTIVE: To explore whether the modified E7-HSP70, which has been introduced mutations in two zinc-binding motifs of E7, will eliminate its transformation potential and enhance the immunogenicity of fusion protein and repress E7 containing tumors with a low level of MHC-I molecules to lung metastatic in murine model. METHODS: In this study, we examined the transforming properties of mutant E7 oncoprotein by the soft agar colony-formation assays, explored the immunogenicity of modified E7-HSP70 gene by various cellular and humor immune responses and evaluated the effect of treating lung metastatic tumor with a low expressing MHC-I molecules by tumor challenge assay and therapeutic experiment. RESULTS: The mutant E7 oncoprotein has completely lost its transforming properties as measured in the soft agar colony-formation assays. Modified E7 HSP70 gene inducted stronger E7-specific cellular immune response than that induced by unmodified E7-HSP70. More importantly, the new construct significantly reduced the number of B16-HPV16E7 lung metastases. CONCLUSION: The modified E7 HSP70 gene may be as a powerful and safe DNA vaccine in controlling the hematogenous spread of HPV16E7-associated tumors with low expression of MHC-I molecules. In addition, the B16-HPV16E7 lung metastasis model can be used to test the efficacy of various E7-specific vaccines and immunotherapeutic strategies in settings more relevant to clinical requirements. PMID- 17081599 TI - Comparative aspects of somatic cell nuclear transfer with conventional and zona free method in cattle, horse, pig and sheep. AB - Nuclear transfer (NT) is a complex procedure that requires considerable technical skills. Over the years attempts have been made to simplify the micromanipulations involved and to make the procedure more user-friendly. A significant step forwards has been the development of the zona-free NT methods. We have used zona free NT with mechanical aspiration of the metaphase plate as a mean of enucleation, in a comparative approach with the conventional nuclear transfer zona-enclosed method in cattle, horse, sheep and pig. The absence of the zona considerably facilitates the enucleation step and significantly increases cell fusion success. On the other hand, the culture of zona-free NT embryos requires the embryos to be cultured individually or anyway separated from each other to avoid aggregation and also requires to prolong the in vitro culture up to the blastocyst stage before transfer. Blastocyst rate is equal or higher with zona free method as compared to zona-enclosed method while survival after cryopreservation and development to term is comparable. In conclusion, our findings, together with published data, demonstrate that the zona-free system described in this paper can significantly increase the output of NT blastocysts over the conventional zona-enclosed system. PMID- 17081600 TI - Dietary transfer of fluoranthene from an estuarine oligochaete (Monopylephorus rubroniveus) to grass shrimp (Palaemonetes pugio): Influence of piperonyl butoxide. AB - The objective of this study was to determine the potential for dietary transfer of sediment-associated fluoranthene from tubificid oligochaetes (Monopylephorus rubroniveus) to grass shrimp (Palaemonetes pugio). Grass shrimp, either in the presence or absence of sublethal waterborne concentrations of the metabolic inhibitor, piperonyl butoxide (PBO), were fed fluoranthene-dosed oligochaetes for 5-days. All grass shrimp bioaccumulated fluoranthene; however, bioaccumulation was 3X higher in the presence of PBO. Trophic transfer coefficients (TTCs) were 0.02 and 0.01 in the presence and absence of PBO, respectively. Following the 5 day accumulation period, shrimp in both treatments were allowed to depurate for 3 days. Depuration rates were significantly higher in PBO-exposed shrimp. These results demonstrated that sediment-associated fluoranthene can be transferred through the diet from oligochaetes to grass shrimp, and the presence of PBO enhanced fluoranthene bioaccumulation. However, the comparatively low TTCs suggest that biomagnification of fluoranthene in estuarine food webs is low. PMID- 17081601 TI - Mesenchymal stem cell ingrowth and differentiation on coralline hydroxyapatite scaffolds. AB - Culture of osteogenic cells on a porous scaffold could offer a new solution to bone grafting using autologous human mesenchymal stem cells (hMSC) from the patient. We compared coralline hydroxyapatite scaffolds with pore sizes of 200 and 500 microm for expansion and differentiation of hMSCs. We cultivated the hMSC statically or in spinner flasks for 1, 7, 14 and 21 days and found that the 200 microm pore scaffolds exhibited a faster rate of osteogenic differentiation than did the 500-microm pore scaffolds as shown by an alkaline phosphatase activity assay and real-time reverse transcriptase polymerase chain reaction for 10 osteogenic markers. The 500-microm scaffolds had increased proliferation rates and accommodated a higher number of cells (shown by DNA content, scanning electron microscopy and fluorescence microscopy). Thus the porosity of a 3D microporous biomaterial may be used to steer hMSC in a particular direction. We found that dynamic spinner flask cultivation of hMSC/scaffold constructs resulted in increased proliferation, differentiation and distribution of cells in scaffolds. Therefore, spinner flask cultivation is an easy-to-use inexpensive system for cultivating hMSCs on small to intermediate size 3D scaffolds. PMID- 17081602 TI - Combinatorial protein display for the cell-based screening of biomaterials that direct neural stem cell differentiation. AB - Neural stem cell (NSC) has emerged as a potential source for cell replacement therapy following traumatic injuries and degenerative diseases of the central nervous system. However, clinical applications of NSC further require technological advances especially for controlling differentiation of NSC. This study aimed at developing biomaterials that serve to expand undifferentiated NSC or to induce cells with specific phenotypes. Our approach is to construct composite biomaterials that consist of extracellular matrix components and growth factors. In order to optimize matrix-growth factor combinations, we conducted the parallel and rapid screening of composite biomaterials through assays using cell based arrays. The photo-assisted patterning of an alkanethiol self-assembled monolayer was employed to achieve site-addressable combinatorial immobilization of natural and synthetic matrices incorporated with growth factors including epidermal growth factor (EGF), ciliary neurotrophic factor (CNTF), nerve growth factor (NGF), and neurotrophin-3 (NT-3). NSC obtained from the rat embryonic striatum was cultured directly on the array to screen for cell adhesion, proliferation, and promotion of neuronal and glial specification. The results showed that the significant number of cells adhered to laminin-1, fibronectin, ProNectin, and poly(ethyleneimine). It was found that cells proliferated most extensively on a spot with immobilized EGF among the spots with different matrix growth factor combinations. The results also showed that neuronal differentiation was promoted on the spots with immobilized NGF or NT-3, and astroglial differentiation with CNTF. Importantly, observed effects of growth factors were frequently altered depending on the type of co-immobilized matrices, suggesting synergic effects of adhesion and growth factor signals. PMID- 17081603 TI - Primary hepatocyte survival on non-integrin-recognizable matrices without the activation of Akt signaling. AB - The suppression of the detachment-induced cell death (anoikis) by the interaction between the cells and extracellular matrix (ECM) is necessary for the application of liver tissue engineering because the disruption of interaction with ECM leads hepatocytes to anoikis. It has been considered, in general, that integrin signal plays an important role in the hepatocyte survival although hepatocytes survive on some types of non-integrin-recognizable matrices, such as poly(N-p-vinylbenzyl 4-O-beta-D-galactopyranosyl-D-gluconamide) (PVLA) and poly-L-lysine (PLL) for several days without the serum. Anoikis was suppressed in the non-adherent culture of hepatocytes isolated from gld/gld mouse, indicating that Fas signal induces hepatocyte anoikis. Fas production is decreased in the adherent culture of hepatocytes on both integrin- and non-integrin-recognizable matrices. Akt activation was hardly observed in the adherent culture of hepatocytes on non integrin-recognizable matrices whereas the activation occurred in the adherent culture on integrin-recognizable matrices. In the adherent culture of hepatocytes on non-integrin-recognizable matrices, Akt does not contribute to the hepatocyte survival. To prolong the viability of hepatocytes in the adherent culture on PVLA matrix on which hepatocytes maintain their functions for longer period than those on PLL matrix, it might be a good approach to activate Akt signaling pathway. PMID- 17081604 TI - Esophageal epithelium regeneration on fibronectin grafted poly(L-lactide-co caprolactone) (PLLC) nanofiber scaffold. AB - In order to mimic normal epithelium regeneration on synthetic scaffold in vitro, biodegradable elastic poly(l-lactide-co-caprolactone) (PLLC) was processed into nanofibrous scaffold using electrospinning technology. An adhesive protein, fibronectin (Fn), was grafted onto the scaffold fiber surface via a two-step reaction: polyester aminolysis followed by Fn coupling via glutaraldehyde. Tensile testing was performed to measure the effect of aminolysis on the scaffold mechanical properties. The strain decreased but the tensile strength remained almost constant after aminolysis. However, no obvious difference of the nanofiber surface morphology was found after Fn grafting using scanning electron microscopy (SEM). Porcine esophageal epithelial cells were seeded on the Fn bonded scaffold to test the cell growth promotion against the control unmodified PLLC nanofiber scaffold using tissue culture polystyrene (TCPS) plate as a reference. Anti cytokeratin AE1/AE3 was used as the primary antibody to confirm the esophageal epithelial phenotype. SEM observation, immunostaining and Western Blotting to compare the collagen type IV synthesis showed that the Fn grafted on PLLC scaffold greatly promotes epithelium regeneration. This modified scaffold is expected to be a good candidate for functional esophagus substitutes. PMID- 17081605 TI - Longitudinal blood flow in shared (arteriovenous anastomoses) and non-shared cotyledons in monochorionic placentae. AB - OBJECTIVES: In this study we aimed to quantitate monochorionic twin placental blood flow in vivo through arterio-venous anastomoses (AVA) and corresponding vessels within normal cotyledons. METHODS: The topography of chorionic plate vasculature was mapped using colour Doppler in ten monochorionic diamniotic twin (MCDA) pregnancies. Cotyledonary flow was derived by insonation of chorionic veins draining normal (n=10) and paired control shared cotyledons (n=10). Venous volume flow was calculated from five determinations of vessel diameter and three of time average mean velocity (TAMV). Measurements were repeated every 2-4 weeks from 18 until 32 weeks' gestation. RESULTS: Blood flow through non-shared and shared cotyledons increased with gestation (p<0.0001). Median flow at 28 weeks through shared cotyledons was 16 ml/min (15-21) (median, interquartile range), lower than in shared cotyledons (31, 25-35) (p<0.001), as was median volume flow across gestation calculated as area under the curve (shared cotyledons 126 (122 167), control cotyledons 269 (214-274), p=0.01). However, velocity was similar, with the difference due to smaller vein diameters draining shared compared to normal cotyledons (mean 3.6mm (SD 0.8) vs. 4.5mm (0.8), p=0.004). Ex vivo quantitation of insonated cotyledons and of all cotyledons confirmed the difference in vein diameter in the placentae studied. CONCLUSIONS: Blood flow through shared cotyledons was lower across gestation than through paired normal cotyledons in the placenta studied due to the smaller diameter of the AVA vessels. The size of AVAs rather than simply their presence and direction may contribute to determining transfusional imbalance in monochorionic twins. PMID- 17081606 TI - Inhibition of HuR and MMP-9 expression in macrophage-differentiated HL-60 myeloid leukemia cells by green tea polyphenol EGCg. AB - Matrix metalloproteinase (MMP)-9 expression is linked with myeloid cell differentiation, as well as inflammation and angiogenesis processes related to cancer progression. MMP-9 secretion and macrophage-like HL-60 myeloid leukemia cells differentiation were triggered by the tumor-promoting agent PMA. The chemopreventive effects of green tea catechins epigallocatechin-gallate, catechin gallate, and epicatechin-gallate, but not those catechins that lack a 3'-galloyl group, inhibited in a time- and dose-dependent manner MMP-9 secretion. The gene and protein expression of MMP-9 and of the mRNA stabilizing factor HuR were also inhibited, while that of the 67 kDa laminin receptor remained unaffected. Specific catechins may help optimize current chemotherapeutic treatment protocols for leukemia. PMID- 17081607 TI - Concentration of organochlorine in egg yolk and reproductive success of Egretta garzetta (Linnaeus, 1758) at Wat Tan-en non-hunting area, Phra Nakhorn Si Ayuthaya Province, Thailand. AB - Contaminant analyses in animal eggs have illustrated the benefit of wildlife as indicators of xenobiotic contamination in natural habitats. In 2002, concentrations of 16 organochlorine insecticides were measured in egg yolk and the reproductive success of the little egret, Egretta garzetta, was studied in a population at Wat Tan-en non-hunting area Bang Pahan District, Phra Nakhorn Si Ayuthaya Province, Thailand. Only 4,4'-dichloro diphenylethane (4,4'-DDE) was detected in the yolk of all samples (n=12) at the concentrations ranging from 33.4 to 116.0 ng/gwetwt. Mean eggshell thickness (n=24) was 0.261+/-0.005 mm. Relationships between the concentrations of 4,4'-DDE and eggshell thickness as well as the reproductive success at all stages were not found (P>0.05). Rather, major factors potentially influencing the decline of reproductive success observed during the field study may include strong wind, predators, intraspecific and interspecific competitions. PMID- 17081608 TI - PLC or TLC: is outpatient commitment the/an answer? AB - The lively debate over mandated community treatment in general and outpatient commitment laws (OPC) in particular has raised many issues. At its core, the debate is over how and to what extent laws should be formulated to persuade, leverage or coerce (PLC) persons with severe mental illness living in the community to comply with medications that mental health professionals believe they need. The alternative to PLC is what we call TLC (tender loving care): a strategy of using benefits - improved patient-centered treatment, entitlements and service delivery, including assertive outreach - rather than penalties or conditions on access to services, to induce compliance. We examine three aspects of the debate: (1) the empirical case for the need for OPC court orders to maintain revolving-door severely mentally ill persons in the community; (2) the normative argument over whether such orders constitute coercion, and, if so, whether that coercion is justifiable; and (3) the incentives such orders create to leverage community providers to augment resources and tailor treatment and services to entice patients to become willing participants in the management of their disorders. PMID- 17081609 TI - Comparison of gene expression by co-cultured WC1+ gammadelta and CD4+ alphabeta T cells exhibiting a recall response to bacterial antigen. AB - Immunization of cattle with a Leptospira borgpetersenii serovar hardjo-bovis vaccine results in the development of a recall response by WC1(+) gammadelta T cells and CD4(+) alphabeta T cells characterized by proliferation and interferon gamma production. It was hypothesized that these two T cell subpopulations had largely redundant effector functions, principally differing in their requirements for activation. To test this, gene expression in cells proliferating to antigen were compared utilizing RT-PCR and bovine microarrays. Both T cell populations had similar transcript profiles for effector molecules, including IFN-gamma, FasL and granzyme B. In contrast, transcripts for costimulatory receptors and ligands were notably different following activation, as WC1(+) T cells expressed no or lower levels of transcripts for CD28 and CD40L, while CD4(+) T cells expressed substantial levels of both. However, both cell types had high levels of CTLA-4 transcript suggesting the cells may be regulated similarly following activation but differ in their need for and ability to provide costimulation. Microarray analyses to extend the number of genes examined revealed that while both subpopulations upregulated anti-apoptotic genes as well as those involved in cell activation and protein biosynthesis, overall there were limited differences between the two antigen-activated cell populations. Those genes that did differ were involved in cell signaling, protein production and intracellular protein trafficking. These results strengthen the hypothesis that these particular activated WC1(+) and CD4(+) T cells have overlapping effector functions and therefore may differ principally with regard to how they are recruited into immune responses. PMID- 17081610 TI - Chemokine-mediated inflammation: Identification of a possible regulatory role for CCR2. AB - The chemokine receptor CCR2 binds four pro-inflammatory monocyte chemoattractant proteins, designated MCP1/CCL2, MCP2/CCL8, MCP3/CCL7 and MCP4/CCL13. This study demonstrates the important biology of this receptor during the response to the chemokine milieu. Competitive chemotaxis and calcium flux assays were performed utilising mixtures of chemokines to assess a hierarchal arrangement of chemokine prepotency; these demonstrated that the MCP2-CCR2 interaction is able to supersede signals generated by RANTES, another pro-inflammatory chemokine, or the homeostatic chemokine SDF1. These observations were validated using three physiologically relevant monocytic cell lines. Having identified the importance of CCR2, experiments were then performed to examine the signal transduction processes coupled to this receptor. G protein coupling was initially examined; Cholera toxin reduced the chemotactic response to MCP2 (p<0.001), whilst the response to the other MCP chemokines remained normal. The response to MCP2 was uniquely inhibited by elevated concentrations of cAMP and, unlike MCP1, 3 and 4 (p<0.05), MCP2 failed to inhibit adenylate cyclase. Expression of dominant negative H-ras demonstrated that each MCP chemokine required active ras in order to elicit ERK activation and a chemotactic response. Unlike MCP1, MCP2 failed to induce nuclear translocation of activated ERK1 or subsequent induction of c-Myc expression. Akt activation also showed ligand-specific differences, with MCP2 producing a delayed response compared to the other MCP chemokines. Together these data highlight the importance of CCR2 and suggest that it is a powerful tool for fine tuning the immune response. PMID- 17081611 TI - Human parotid saliva contains soluble toll-like receptor (TLR) 2 and modulates TLR2-mediated interleukin-8 production by monocytic cells. AB - Toll-like receptor (TLR) family members are pattern-recognition receptors and very important molecules in innate immunity. Although TLRs are originally type I transmembrane receptors, soluble forms of TLRs are detected in human plasma and milk. This study showed that soluble TLR2 (sTLR2) is detected in human parotid saliva. Western blotting with anti-TLR2 antibodies (Abs) showed that three polypeptides are detected as sTLR2 with molecular weights of 55, 40 and 27kDa, respectively. Parotid saliva neutralized the binding of anti-TLR2 polyclonal Ab to cell-surface TLR2 on THP-1, a human monocytic cell line. Immunohistochemical analysis revealed that TLR2 is expressed in serous and interlobular ductal cells of human salivary gland. Human salivary gland cell lines, AZA3 and HSY, constitutively expressed TLR2. Parotid saliva augmented IL-8 production of THP-1 cells stimulated with a synthetic TLR2 ligand, Pam(3)Cys-Ser-(Lys)(4) (Pam(3)CSK(4)). Depletion of sCD14 from parotid saliva by immunoprecipitation eliminated the augmentation of IL-8 production, indicating that the augmentable effects depended on sCD14 in parotid saliva. On the other hand, preincubation of Pam(3)CSK(4) with parotid saliva abrogated the augmentation of IL-8 production, indicating that sTLR2 in saliva bound to Pam(3)CSK(4) and neutralized its function. These results suggest that parotid saliva modulates the TLR2-mediated immune responses with binary mechanisms via sTLR2 and sCD14 in the oral cavity. PMID- 17081612 TI - When being straight bends rules: a rationale for the linear FeNO unit in the low spin square-pyramidal {FeNO}7 tetracyanonitrosylferrate(2-) anion. AB - All low-spin S=1/2 heme-NO complexes feature FeNO angles of about 140 degrees . In contrast, the square-pyramidal [Fe(CN)(4)(NO)](2-) complex features an exactly linear {FeNO}(7) unit. We have sought here to determine a possible, simple molecular orbital (MO) rationale for these structural variations. A DFT-based (DFT=density functional theory) MO analysis shows that the linearity of the latter stems from the greater pyramidalization of the Fe center, relative to nitrosylheme, which results in significant differences in d orbital hybridization. Thus, the singly occupied molecular orbital (SOMO) of [Fe(CN)(4)(NO)](2-) , while primarily Fe dz2-based, also has a significant amount of 4p(z) character, which makes it less stereochemically active, accounting for the linearity of the FeNO unit. PMID- 17081613 TI - The human embryonic stem cell-derived cardiomyocyte as a pharmacological model. AB - Embryonic stem (ES) cells are specialised cells derived from the early embryo, which are capable of both sustained propagation in the undifferentiated state as well as subsequent differentiation into the majority of cell lineages. Human ES cells are being developed for clinical tissue repair, but a number of problems must be addressed before this becomes a reality. However, they also have potential for translational benefit through its use as a test system for screening pharmaceutical compounds. In the cardiac field, present model systems are not ideal for either screening or basic pharmacological/physiological studies. Cardiomyocytes produced from human ES differentiation have advantages for these purposes over the primary isolated cells or the small number of cell lines available. This review describes the methodology for obtaining cardiomyocytes from human embryonic stem cell-derived cardiomyocyte (hESCM), for increasing the proportion of cardiomyocytes in the preparation and for isolating single embryonic stem cell-derived cardiomyocyte (ESCM) from clusters. Their morphological, contractile and electrophysiological characteristics are compared to mature and immature primary cardiomyocytes. The advantages and disadvantages of the hESCM preparation for long term culture and genetic manipulation are described. Basic pharmacological studies on adrenoceptors and muscarinic receptors in hESCM have been performed, and have given stable and reproducible responses. Prolongation of repolarisation can be detected using hESCM cultured on multielectrode arrays (MEA). Human ESCM have a clear potential to improve model systems available for both basic scientific studies and pharmaceutical screening of cardiac target compounds. PMID- 17081614 TI - Ultrasound-assisted digestion: a useful alternative in sample preparation. AB - Ultrasound-assisted digestion is a promising alternative in the analysis of solid samples when either simple dissolution or direct analysis is not applicable. However, the field of application of ultrasonic sample digestion is still small in comparison with classical digestion alternatives and, particularly, with microwave-assisted digestion. This fact can be justified by the scant knowledge analytical chemists have about the advantages ultrasonic energy provides to digestion. Among these, the strict control at low temperatures of ultrasound applications allows the implementation of ultrasonic-assisted steps in biochemical analyses. In this connection, two specific biological applications, ultrasonic enzymatic digestion and assistance of ultrasound for cell disruption, are also reviewed. PMID- 17081615 TI - Amperometric hydrogen peroxide biosensor based on the immobilization of HRP on nano-Au/Thi/poly (p-aminobenzene sulfonic acid)-modified glassy carbon electrode. AB - A novel hydrogen peroxide biosensor was fabricated for the determination of H(2)O(2). The precursor film was first electropolymerized on the glassy carbon electrode with p-aminobenzene sulfonic acid (p-ABSA) by cyclic voltammetry (CV). Then thionine (Thi) was adsorbed to the film to form a composite membrane, which yielded an interface containing amine groups to assemble gold nanoparticles (nano Au) layer for immobilization of horseradish peroxidase (HRP). The electrochemical characteristics of the biosensor were studied by CV and chronoamperometry. The factors influencing the performance of the resulting biosensor were studied in detail. The biosensor responded to H(2)O(2) in the linear range from 2.6 x 10(-6) mol/L to 8.8 x 10(-3) mol/L with a detection limit of 6.4 x 10(-7) mol/L. Moreover, the studied biosensor exhibited good accuracy and high sensitivity. The proposed method was economical and efficient, making it potentially attractive for the application to real sample analysis. PMID- 17081616 TI - Developments in multiple headspace extraction. AB - This paper reviews new developments in multiple headspace extraction (MHE), especially its combination with two miniaturized extraction techniques, solid phase microextraction (SPME) and single-drop microextraction (SDME). The combination of the techniques broadens the applicability of SPME and SDME to quantitative determination of analytes in complex liquid and solid matrixes. These new methods offer several advantages over traditional liquid-solid, liquid liquid and headspace extraction techniques. The potential applications include extraction of volatiles and semivolatiles from environmental and physiological samples and from different polymer products such as medical and biomedical materials, food packaging and building materials. The theoretical principals of the techniques are also briefly reviewed. PMID- 17081617 TI - Study of hypothermia on cultured neuronal networks using multi-electrode arrays. AB - Efficient and safe use of hypothermia during various neuro-medical procedures requires sound understanding of low temperature effects on the neuronal network's activity. In this report, we introduce the use of cultivated dissociated neuronal networks on temperature controlled multi-electrode arrays (MEAs) as a simple methodology for studying the long-term effects of hypothermia. The networks exhibit spontaneous activity in the form of synchronized bursting events (SBEs), followed by long intervals of sporadic firing. Through the use of our correlation method, these SBEs can be clustered into sub-groups of similar spatio-temporal patterns. Application of hypothermia to the network resulted in a reduction in the SBE rate, the spike intensity and an increase in inter-neuronal correlations. Within 2h following the cessation of hypothermia, the cultured network returned to its initial spatio-temporal SBE structure. These results suggest that the network survived cold exposure and demonstrate the feasibility of long-term continuous neural network recording during hypothermic conditions. PMID- 17081618 TI - Methods for building an olfactometer with known concentration outcomes. AB - We provide detailed instructions and part selections for construction of a five channel air dilution olfactometer capable of generating neat odorants and binary mixtures at a range of known concentrations. At the heart of the olfactometer is an odorant canister that is (1) cheap and readily available, (2) safe and durable, (3) has minimal odor adherence, (4) is easily incorporated into any olfactometer, and critically (5) produces a highly consistent stimulus. By flowing a given carrier gas at a given flowrate through a given odorant in this canister, the same end-vapor is achieved. Flow/concentration outcomes are provided for several odorants routinely used in olfactometry. This tool will enable researchers to generate known concentrations without expensive analytical machinery. PMID- 17081619 TI - MHC expression and chemokine production in the murine vagina following intra vaginal administration of ligands to toll-like receptors 3, 7 and 9. AB - The expression of MHC class I, MHC class II and the chemokines IP-10, MIP-1alpha, RANTES, fractalkine and I-TAC has been analyzed after intra-vaginal treatment with three synthetic toll-like receptors (TLR) agonists-double-stranded RNA (poly I:C), imiquimod and CpG-rich oligonucleotides (CpG-ODN). These compounds act mainly through TLR3, TLR7 and TLR9, respectively. CpG-ODN induced an accumulation of leucocytes in the vagina, and a strong up-regulation of MHC class I expression on both leucocytes and epithelial cells. Imiquimod and poly I:C induced a weak MHC class I up-regulation in the epithelium but not in the lamina propria. Neither treatment had any profound effect on expression of MHC class II on epithelial cells but poly I:C and to a lesser extent CpG-ODN, up-regulated MHC class II staining intensity which, in the case of CpG-ODN, treatment, was associated with a strong accumulation of CD11c-positive dendritic cells. All three treatments induced an early (8h) but transient IP-10 response. Imiquimod and CpG-ODN, but not poly I:C induced an early MIP-1alpha response which remained for at least 7 days in CpG-ODN-treated animals but not in imiquimod-treated mice. Poly I:C and CpG-ODN, but not imiquimod, induced significant levels of RANTES at different time-points post-treatment. None of the treatments induced any significant changes in the levels of fractalkine, I-TAC or IFN-alpha. These studies have implications for the manipulation of the genital immune response and also improving the outcome of vaginal immunotherapy. PMID- 17081620 TI - Perceptual grouping in disorganized schizophrenia. AB - This study evaluated visual perceptual grouping in schizophrenia to test the hypothesis that the disorganization syndrome in schizophrenia is related to a deficit in cognitive coordination. Perceptual grouping was examined with three psychophysically well-controlled tasks in patients with disorganized schizophrenia (n=11), non-disorganized schizophrenia (n=24), psychotic disorders other than schizophrenia (n=31) and non-psychotic psychiatric disorders (n=35). These measures assessed processing of both concurrent and preceding stimulus context. Deficits in perceptual grouping were observed on all three tasks in disorganized schizophrenia patients. Dysfunctional perceptual grouping mechanisms produced both enhanced and impaired task performance suggesting that the pattern of performance observed was the result of a specific deficit in the grouping of stimulus elements. We interpret these data as further support for the hypothesis that the disorganization syndrome in schizophrenia reflects a widespread deficit in the cognitive coordination of contextually related stimuli, leading to dysfunctional grouping of stimulus features in vision, thought and language. PMID- 17081621 TI - Transient hypofrontality as a mechanism for the psychological effects of exercise. AB - Although exercise is known to promote mental health, a satisfactory understanding of the mechanism underlying this phenomenon has not yet been achieved. A new mechanism is proposed that is based on established concepts in cognitive psychology and the neurosciences as well as recent empirical work on the functional neuroanatomy of higher mental processes. Building on the fundamental principle that processing in the brain is competitive and the fact that the brain has finite metabolic resources, the transient hypofrontality hypothesis suggests that during exercise the extensive neural activation required to run motor patterns, assimilate sensory inputs, and coordinate autonomic regulation results in a concomitant transient decrease of neural activity in brain structures, such as the prefrontal cortex, that are not pertinent to performing the exercise. An exercise-induced state of frontal hypofunction can provide a coherent account of the influences of exercise on emotion and cognition. The new hypothesis is proposed primarily on the strength of its heuristic value, as it suggests several new avenues of research. PMID- 17081622 TI - Anti-OxLDL IgG blocks OxLDL interaction with CD36, but promotes FcgammaR, CD32A dependent inflammatory cell adhesion. AB - Generation of antibodies against oxidized-low-density lipoproteins (oxLDL) during atherosclerosis could result in the formation and deposition of oxLDL immune complexes (oxLDL-IC) on the vascular endothelial cells. Inflammatory cells express scavenger receptor (SR such as CD36) and Fcgamma receptor (FcgammaR: CD32A and CD64) that can bind to oxLDL and oxLDL-IC, respectively. Hence, depending on anti-oxLDL IgG titer, circulating monocytes could adhere to endothelium to oxLDL-IC-coated vascular bed via either FcgammaR and/or CD36. In this study, we determined the relative contribution of SR and FcgammaR in mediating monocyte interaction with oxLDL-IC deposited on vascular bed. At saturating levels of anti-oxLDL IgG concentration, monocytic cells adhered to oxLDL-IC and this adhesion is completely blocked by anti-CD32A mAb. Using CHOK1 CD32A-CD36 cells expressing equal levels of CD32A and CD36, it was observed that at lower concentrations of anti-oxLDL IgG, CD32A and CD36 contribute about 75% and 25% of cell adhesion, respectively, while at higher concentrations of anti oxLDL IgG the adhesion is completely CD32A-dependent. CD32A-dependent adhesion was further confirmed with peripheral blood monocytes and platelets that express 2- to 5-fold higher levels of CD36 compared to CD32A. Further, PBMC adhesion to oxLDL-IC-deposited endothelial cells induced secretion of pro-inflammatory chemokines, MCP-1 and IL-8. Our results demonstrate that anti-oxLDL IgG blocks oxLDL interaction with SR such as CD36, whereas oxLDL-IC formation promotes monocyte adhesion and subsequent chemokine release through FcgammaR. These findings suggest a role for FcgammaR-mediated inflammatory cell activation in the progression of atherosclerosis. PMID- 17081623 TI - CD4(+) memory T cells with high CD26 surface expression are enriched for Th1 markers and correlate with clinical severity of multiple sclerosis. AB - An aberrant immune activation is believed to be important in the pathogenesis of multiple sclerosis (MS). Expression of CD4(+) T lymphocyte surface molecules indicative of immune activation and effector functions has been correlated with disease severity and activity. CD4(+) CD45R0(+) CD26(high) memory T lymphocytes contained the high levels of markers of Th1, activation, and effector functions and cell counts of this subset correlated with MS disease severity. This subset had lower expression of PD-1, CCR4, and L-selectin in MS than in controls. These changes were only partially normalised by treatment with interferon-beta. We point to this subset as a putative target for immunological monitoring of MS disease activity and of treatment efficacy. PMID- 17081624 TI - Regulatory effects of vasoactive intestinal peptide on the migration of mature dendritic cells. AB - The neuropeptide vasoactive intestinal peptide (VIP), released within lymphoid organs from nerve terminals and/or immune cells, plays a significant anti inflammatory role. It was reported that VIP can induce regulatory dendritic cells (DCs) and promote Th2-type responses. However, the regulatory effect of VIP on the migration and expression of chemokine receptors by DC is mostly unknown. In the present study, we show that VIP exerts a differential effect on the expression of CCR1 and CCR7 by lipopolysaccharide (LPS)-treated mature DCs (mDCs) at both protein and mRNA levels. It up-regulates CCR1 expression but down regulates CCR7 expression in LPS-stimulated mature DC, thereby differentially regulating the migration of mature DCs in response to CCL5 and CCL19. Our data indicate that VIP functions as a key endogenous anti-inflammatory agent by inhibiting migration of mDCs to draining lymph nodes, thus preventing the induction of an inflammatory immune response. PMID- 17081625 TI - Combined retrograde and anterograde hypopharyngeal puncture and dilatation in a child with complete hypopharyngeal stenosis. AB - The management of complete hypopharyngeal stenosis is a complex task, and various methods of re-creating a lumen and maintaining its patency have been described in both the adult and pediatric literature. We present our experience using a modification of the combined anterograde-retrograde endoscopic technique for managing a 15-year-old child who developed complete hypopharyngeal and upper esophageal obliteration following successful treatment of a parapharyngeal rhabdomyosarcoma. During an 18-month period following initial cannulation of the aerodigestive tract, we performed 17 dilatations using a double-balloon technique that combined anterograde and retrograde approaches to the stenosis. Initially, Kenalog injections and topical Mitomycin-C were used as adjunctive treatments. We observed no major complications following dilatations. At 33 months follow-up the patient was asymptomatic, tolerating a near normal diet, had not required dilation for more than 18 months, and no longer required a gastrostomy tube. PMID- 17081626 TI - Polyflex stenting of tracheomalacia after surgery for congenital tracheal stenosis. AB - Polyflex self-expanding stents (Rusch, Germany) were used in three young children who had presented with life-threatening long-segment tracheal stenosis with bronchial stenosis in two cases. Two children had slide tracheoplasties and subsequently aortic homografts and another tracheal resection and autotracheoplasty. However, in all cases persistent lower tracheal malacia necessitated stenting. Complications of granuloma, stent migration or dislodgement occurred in all cases. A fatal tracheo-aortic fistula occurred in one child. Granuloma in one was treated successfully with steroids. One child survives. PMID- 17081627 TI - Bacteriophage T4 nanoparticle capsid surface SOC and HOC bipartite display with enhanced classical swine fever virus immunogenicity: a powerful immunological approach. AB - The phage T4 HOC, SOC bipartite display system is attractive for the expression of cDNA and display of peptides or proteins at high copy numbers on the phage capsid surface. Until recently, using T4 phage vector to display large foreign molecular immunogens resulted only from either an SOC or HOC single site. In this report, the main advantages of the phage T4 system over other display technologies are substantiated by using the phage T4 SOC, HOC dual site display vector T4-Zh(-) to express: (1) on the SOC site, the classical swine fever virus (CSFV) major antigenic determinant cluster mE2 (123 amino acid, aa) through gene fusion to the SOC gene C-terminus of T4 genome, and (2) on the HOC site, full length CSFV primary antigen E2 (371 aa) through a co-transformed plasmid, hence leading to a simultaneous display of both proteins on the T4 capsid surface. The immunogenicities of these constructs were measured by ID-ELISA, dot-ELISA, Western blotting, and immunogenic response in mice including humoral and cellular immunity tests. The immunological efficiencies both in vitro and in mice of phage T4 with both single site and dual site displays, as well as conventional Escherichia coli plasmid expression, were evaluated. The animal immune response data showed that the antibody titers elicited by the T4 phage-CSFV recombinants were significantly higher than those obtained by E. coli plasmid expression, and the unpurified double site display T4 phage particles were around two times higher than either single site display or plasmid expression while being at lower phage concentrations than the single site phages. The immunogens were effective in the absence of eukaryotic protein modifications. Therefore, the phage T4 dual site display emerges as a powerful method with an enhanced immune response in animals for research and development of immunological products. PMID- 17081628 TI - Central nicotinic cholinergic systems: a role in the cognitive dysfunction in attention-deficit/hyperactivity disorder? AB - Theories of the neurobiological basis of Attention-Deficit/Hyperactivity Disorder (ADHD) have largely focused on dysregulation of central dopaminergic function. However, other neurotransmitter systems may be implicated in specific cognitive deficits in ADHD. Interest in the potential involvement of nicotinic cholinergic systems in ADHD has arisen in part from the observation that adolescents and adults with ADHD smoke cigarettes at significantly higher rates than people without this disorder. In addition, several studies report that nicotine alleviates ADHD symptoms, and recent neuro-genetics studies indicate that cholinergic systems may be altered in persons with ADHD. In this review, we describe the evidence for a role of central nicotinic cholinergic systems in cognitive deficits in ADHD. We also propose mechanisms by which alterations in cholinergic function may contribute directly and/or indirectly to these deficits. Finally, we identify specific paradigms and models to guide future investigations into the specific involvement of nicotinic cholinergic systems in ADHD, possibly leading to the development of more effective pharmacotherapies for ADHD. PMID- 17081629 TI - Early deprivation, but not maternal separation, attenuates rise in corticosterone levels after exposure to a novel environment in both juvenile and adult female rats. AB - Separation from the maternal nest alters the hypothalamic-pituitary-adrenal (HPA) axis stress response in adult male rats, but little research has addressed how separation affects female rats. The following experiments investigated how early maternal separation from postnatal day (PND) 2 to 14 affected stress-induced corticosterone and ACTH after exposure to an open field in juvenile and adult female rats. Female rats were separated for 5 h daily from mother and littermates (early deprivation: ED), separated from mother but not littermates (maternal separation: MS), or animal facility reared (AFR). Male siblings were left with the mother rat during separation. Female rats were exposed to an open field arena either during the juvenile period (PND 30) or during adulthood (PND 80-100). Results show that ED juvenile female rats showed a lower corticosterone stress response than MS and AFR female rats when measured at 5 min post-stress, but no difference at 20 or 60 min post-stress. In adulthood, ED female rats showed comparable elevations of corticosterone as MS and AFR rats at 5 min post-stress but lower elevations at 20 min. In terms of behavior, there were no significant effects of early experience. However, in adulthood, ED and MS rats tended to show a decreased proportion of inner grid crossings of the open field compared to AFR rats, suggesting a tendency for increased anxiety in these two separation groups. PMID- 17081630 TI - Longitudinal study of the effects of a high-fat diet on glucose regulation, hippocampal function, and cerebral insulin sensitivity in C57BL/6 mice. AB - Although the increasing rate of obesity has stimulated interest in the effects of diet composition on peripheral systems, comparatively little work has been done to examine effects upon the brain. A diet high in fat is one of many factors that can promote obesity, and previous research has shown that such a diet can produce learning and memory impairment in rodents. In the present study, C57BL/6 mice were placed on either a high-fat (45% kcal fat) or regular (5% kcal fat) diet, and examined at different points during the subsequent year. The high-fat diet led to increased weight gain, significant impairment in glucoregulation, and altered insulin-mediated signaling within the hippocampus, an area of the brain believed to be important for the acquisition of memory. Following ten months on either diet, synaptic function in ex vivo hippocampal slices was examined, and neither stimulus-response curves nor electrically induced long-term potentiation were found to be different. As well, performance in the Morris water maze, a hippocampal-dependent test of spatial memory, was not influenced by diet. However, mice consuming a high-fat diet failed to perform an operant bar-pressing task, indicating a significant impairment to procedural learning and consolidation processes. Despite causing broad peripheral changes in C57BL/6 mice, consuming a large proportion of calories from saturated fat had only a limited effect upon learning and memory, which suggests that certain aspects of brain function are selectively vulnerable to the influences of diet. PMID- 17081631 TI - Sensitivity of Atlantic salmon (Salmo salar) to dietary endosulfan as assessed by haematology, blood biochemistry, and growth parameters. AB - The presence of the organochlorine pesticide endosulfan in the water column confers a significant direct risk to the biota therein, yet relatively little is known regarding the toxic impact of dietborne endosulfan to aquatic organisms. Pre-smolt Atlantic salmon (Salmo salar) were fed at levels of technical endosulfan equal to the European Union regulatory limit (5 microg kg(-1)) and at levels 10 and 100 times greater, for a total of 49 days with haematology (e.g. erythrocyte count, haemoglobin, haematocrit, white blood cell composition), blood biochemistry (e.g. serum aminotransferase, plasma ions) and growth parameters (e.g. condition factor) recorded at days 0, 14, 35, and 49. Toxicological assessment of the individual alpha and beta-isomers that comprise technical endosulfan was also attempted. No mortality was observed in any group during the experiment. In the groups exposed to the control, 5, and 50 microg kg(-1) technical endosulfan feeds no significant alterations in any measured parameters were determined at any time point. No differences were observed between the technical mixture and the individual alpha and beta-isomer treatment groups. Condition factor was significantly reduced in fish exposed to 500 microg kg(-1) at day 49, while haematocrit, haemoglobin and mean corpuscular haemoglobin were significantly elevated after 35 days of exposure to the same dietary level, but returned to control levels by day 49. The present study shows that with regards to acute toxicity, Atlantic salmon are able to tolerate dietary technical endosulfan levels up to 500 microg kg(-1). PMID- 17081632 TI - Secretion and processing of a novel multi-domain cystatin-like protein by intracellular stages of Trichinella spiralis. AB - The excretory-secretory (ES) proteins of nematode parasites are of major interest as they function at the host-parasite interface and are likely to have roles crucial for successful parasitism. Furthermore, the ES proteins of intracellular nematodes such as Trichinella spiralis may also function to regulate gene expression in the host cell. In a recent proteomic analysis we identified a novel secreted cystatin-like protein from T. spiralis L1 muscle larva. Here we show that the protein, MCD-1 (multi-cystatin-like domain protein 1), contains three repeating cystatin-like domains and analysis of the mcd-1 gene structure suggests that the repeated domains arose from duplication of an ancestral cystatin gene. Cystatins are a diverse group of cysteine protease inhibitors and those secreted by parasitic nematodes are important immuno-modulatory factors. The cystatin superfamily also includes cystatin-like proteins that have no cysteine protease inhibitory activity. A recombinant MCD-1 protein expressed as a GST-fusion protein in Escherichia coli failed to inhibit papain in vitro suggesting that the T. spiralis protein is a new member of the non-inhibitory cystatin-related proteins. MCD-1 secreted from T. spiralis exists as high- and low-molecular weight isoforms and we show that a recombinant MCD-1 protein secreted by HeLa cells undergoes pH-dependent processing that may result in the release of individual cystatin-like domains. Furthermore, we found that mcd-1 gene expression is largely restricted to intracellular stages with the highest levels of expression in the adult worms. It is likely that the major role of the protein is during the intestinal stage of T. spiralis infections. PMID- 17081633 TI - Identification of genes involved in interactions between Biomphalaria glabrata and Schistosoma mansoni by suppression subtractive hybridization. AB - Biomphalaria glabrata is an intermediate snail host for Schistosoma mansoni, a medically important schistosome. In order to identify transcripts involved in snail-schistosome interactions, subtractive cDNA libraries were prepared, using suppression subtractive hybridization (SSH) between a parasite-exposed schistosome-resistant and a susceptible strain of B. glabrata, and also between schistosome-exposed and unexposed snails from the resistant snail line. Separate libraries were made from both haemocytes and the haemopoietic organ. Subtraction was performed in both directions enriching for cDNAs differentially expressed between parasite-exposed resistant and susceptible samples and up or down regulated in the resistant line after challenge. The resulting eight libraries were screened and eight genes, differentially expressed between the haemocytes of resistant and susceptible snail strains, were identified and confirmed with reverse transcriptase PCR, including two transcripts expected to be involved in the stress response mechanism for regulating the damaging oxidative burst pathways involved in cytotoxic killing of the parasite: the iron-storage and immunoregulatory molecule, ferritin, and HtrA2, a serine protease involved in the cellular stress response. Transcripts with elevated levels in the resistant strain, had the same expression patterns in the subtracted libraries and unsubtracted controls; higher levels in exposed resistant snails compared to susceptible ones and down-regulated in exposed compared with unexposed resistant snails. Differential expression of two of the transcripts with no known function from the susceptible strain, was independently confirmed in a repeat exposure experiment. PMID- 17081634 TI - Placental Plasmodium falciparum infection: causes and consequences of in utero sensitization to parasite antigens. AB - Available evidence suggests that, in African populations, systemic blood-dwelling parasitoses of mothers are associated with enhanced susceptibility to infection of their offspring. Thus, children born to mothers with filariasis or schistosomiasis are infected earlier, and offspring of mothers with placental Plasmodium falciparum at delivery, commonly referred to as pregnancy-associated malaria or PAM, are themselves at higher risk of developing parasitaemia during infancy. Since foetal/neonatal antigen-presenting cells (APC) are either immature or provide insufficient costimulatory signals to T cells, thus favouring tolerance induction, it is commonly assumed that soluble parasite components [protein antigens], transferred transplacentally and inducing foetal immune tolerance, are largely, if not exclusively, responsible for these outcomes. Plasmodial asexual blood stage antigen-specific T cells are detectable in as many as two-thirds of all cord blood samples in malaria-endemic countries of sub Saharan Africa, indicating that in utero sensitization may be a common phenomenon during pregnancy in these populations. Parasite antigen-specific T cell responses of neonates born to helminth-infected mothers display a highly skewed Th2-type cytokine pattern, with a prominent role for the regulatory cytokine interleukin (IL)-10. Similarly, the cord blood immune response of those born to mothers identified with on-going PAM is characterised by inducible parasite antigen specific IL-10-producing regulatory T cells that can inhibit both APC HLA expression and Th1-type T cell responses. In contrast, plasmodial antigen specific Th1-type responses, characterised by IFN-gamma production, predominate in cord blood of those born to mothers successfully treated for Pf malaria during gestation, suggesting that the duration and/or the nature of antigen exposure in utero governs the outcome with respect to neonatal immune responses. Aspects of APC function in the context of these differentially modulated responses, whether and how the latter translate into altered susceptibility to Pf infection during infancy, as well as the possible implications for vaccination in early life, are aspects that are discussed in this review. PMID- 17081635 TI - Use of a fluorescent polarization based high throughput assay to identify new calmodulin ligands. AB - In order to develop a fluorescence polarization (FP) assay for calcium binding proteins, a fluorescent peptides based library of 1328 compounds has been synthesized. The use of this library has been validated by setting up a FP-high throughput screening (FP-HTS) assay for calmodulin using the synthetic gene product (synCaM). With this assay, a set of 880 FDA approved compounds was screened. Besides the promazine class, we discovered two new classes of compounds that interact with calmodulin in a calcium dependent manner. One class has compounds with anti-histaminic/spasmolytic activities, and the other one are detergents with antibacterial activities. PMID- 17081636 TI - Activation of human T lymphocytes via integrin signaling induced by RGD disintegrins. AB - Adhesive interactions play important roles in coordinating T cell migration and activation, which are mediated by binding of integrins to RGD motif found on extracellular matrix proteins. Disintegrins, isolated from snake venoms, contain the RGD sequence that confers selectivity to integrin interaction. We have investigated the ability of three RGD-disintegrins, ligands of alpha(5)beta(1) and alpha(v)beta(3), Flavoridin (Fl), Kistrin (Kr) and Echistatin (Ech), in modulating the activation of human T lymphocyte. The disintegrins induced T cell proliferation and CD69 expression. This activation parallels with actin cytoskeleton reorganization and tyrosine phosphorylation. Furthermore, the peptides induced focal adhesion kinase (FAK) and phosphoinositide 3-kinase (PI3K) activation. Finally, RGD-disintegrins were capable of driving NF-kappaB nuclear translocation and c-Fos expression, in a PI3K and ERK1/2 activities dependent manner. This report is the first to show that RGD-disintegrins interact with integrins on human T lymphocyte surface, modulating cell proliferation and activation of specific pathways coupled to integrin receptor. PMID- 17081637 TI - Rare clinical presentation of nonobstructive hypertrophic cardiomyopathy: apical aneurysm with thrombus. AB - Hypertrophic cardiomyopathy patients rarely have left ventricular apical aneurysms without coronary artery disease and the pathophysiological processes responsible for apical wall thinning and aneurysm formation are not known exactly today. Here we present a rare nonobstructive hypertrophic cardiomyopathy case with an apical thrombotic aneurysm. PMID- 17081638 TI - Anomalous circumflex artery: intravascular ultrasound imaging of dynamic compression and "two wire-two vessel" percutaneous intervention. AB - An anomalous origin of the left circumflex is the most common congenital abnormality encountered in coronary angiography and it may selectively predispose to focal accelerated atherosclerotic disease. We report the case of a 54-year-old man who presented with non-ST elevation myocardial infarction and a characteristic retroaortic (proximal) culprit lesion in his anomalous circumflex artery. Intravascular ultrasound images illustrate the dynamic nature of the lumen compression. A "two wires-two vessel" angioplasty technique provided extra support and is recommended to facilitate successful anomalous circumflex percutaneous coronary interventions. PMID- 17081639 TI - Reporting of suspect cases of foot-and-mouth-disease during the 2001 epidemic in the UK, and the herd sensitivity and herd specificity of clinical diagnosis. AB - We described the clinical diagnostic process utilized during the 2001 epidemic of foot-and-mouth-disease in the United Kingdom (UK), and considered it as a series of diagnostic tests. Premises were classified according to these diagnostic-test results and actual disease status, determined by the reference test, which in this case was one or more internationally accepted laboratory tests. The herd level sensitivity (HSe) and herd-level specificity (HSp) of the clinical diagnostic process were calculated directly, relative to these internationally accepted reference tests. In this process, the first diagnostic test was 'routine monitoring', which resulted in the identification of suspect cases based solely on the clinical observations of farmers or veterinarians. 6762 suspect cases were identified, and the test had a HSe of 97.6% (95% C.I.: 96.7, 98.3) and a HSp of 95.2% (95% C.I.: 95.0, 95.3). Suspect cases were then subject to the second diagnostic test, termed 'declaration', which consisted of a review of a description of the clinical signs by government veterinarians. Premises that tested positive became 'clinical cases'. The HSe of this test was 97.1% (95% C.I.: 96.2, 97.9), and the HSp was 90.9% (95% C.I.: 90.1, 91.6). During the epidemic, these tests were combined and applied in series, with an overall HSe of 94.7% (95% C.I.: 93.5, 95.7) and an overall HSp of 99.6% (95% C.I.: 99.5, 99.6). We also examined the effect of a policy shift that prohibited delaying the diagnosis pending laboratory testing where the animals exhibited equivocal clinical signs. PMID- 17081640 TI - Nuclear traffic of influenza virus proteins and ribonucleoprotein complexes. AB - Influenza virus is a negative strand RNA virus and is one of the rare RNA viruses to replicate in the nucleus. The viral RNA is associated with 4 viral proteins to form ribonucleoprotein particles (RNPs). After cell entry the RNPs are dissociated from the viral matrix protein in the low pH of the endosome and are actively imported into the cell nucleus. After translation of viral mRNAs, the proteins necessary for the assembly of new RNPs (the nucleoprotein and the three subunits of the polymerase complex) are also imported into the nucleus. Apart from these four proteins, part of the newly made matrix protein is also imported and the nuclear export protein (NEP) enters the nucleus probably through diffusion. Finally, NS1 also enters the nucleus in order to regulate a number of nuclear processes. The nuclear localization signals on all these viral proteins and their interaction with the cellular transport system are discussed. In the nucleus, the matrix protein binds to the newly assembled RNPs and NEP then binds to the matrix protein. NEP contains the nuclear export signal necessary for transport of the RNPs to the cytoplasm, necessary for the budding of new virus particles. There appears to be a intricate ballet in exposing and hiding nuclear transport signals which leads to a unidirectional transport of the RNPs to the nucleus at the start of the infection process and an opposite unidirectional export of RNPs at the end of the infection. PMID- 17081641 TI - A totivirus infecting the mutualistic fungal endophyte Epichloe festucae. AB - Epichloe festucae (Ascomycota) infects the grass Festuca rubra. Infected plants may be more resistant to herbivores and obtain other benefits. The 5109bp dsRNA genome of a virus which infects E. festucae was sequenced, and its incidence in natural populations and transmission were studied. The viral genome has characteristics of the family Totiviridae. Its two ORFs are overlapped by four nucleotides; ORF1 codes a 765 amino acid putative coat protein (CP); ORF2 is in a -1 frameshift with respect to ORF1, and codes a 826 amino acid RNA dependent RNA polymerase (RdRp). This virus, denominated Epichloe festucae virus 1 (EfV1), is closely related to members of the genus Totivirus which infect filamentous fungi, as deduced from phylogenetic analyses of CPs and RdRps. In two natural populations of Epichloe festucae, 36.4% of the isolates were infected by EfV1. The virus was efficiently transmitted to asexual fungal spores. However, when ascospore progeny of matings between virus-free and infected strains was analyzed, it was found that the virus was not transmitted to progeny of sexual spores. PMID- 17081642 TI - Evaluating the intracellular stability and unpacking of DNA nanocomplexes by quantum dots-FRET. AB - We demonstrate a highly sensitive method to characterize the structural composition and intracellular fate of polymeric DNA nanocomplexes, formed by condensing plasmid DNA with cationic polymers through electrostatic interactions. Rational design of more efficient polymeric gene carriers will be possible only with mechanistic insights of the rate-limiting steps in the non-viral gene transfer process. To characterize the composition and binding dynamics of nanocomplexes, plasmid and its polymer carrier within nanocomplexes were labeled with quantum dots (QDs) and fluorescent organic dyes, respectively, as a donor and acceptor pair for fluorescence resonance energy transfer (FRET). The high signal-to-noise ratio in QD-mediated FRET enabled precise detection of discrete changes in nanocomplex state at the single-particle level, against various intracellular microenvironments. The distribution and unpacking of individual nanocomplexes within cells could thus be unambiguously followed by fluorescence microscopy. QD-FRET is a highly sensitive and quantitative method to determine the composition and dynamic stability of nanocomplexes during intracellular transport, where barriers to gene delivery may be identified to facilitate gene carrier optimization. PMID- 17081643 TI - In vivo evaluation of EPO-secreting cells immobilized in different alginate-PLL microcapsules. AB - Alginates are the most employed biomaterials for cell encapsulation due to their abundance, easy gelling properties and apparent biocompatibility. However, as natural polymers different impurities including endotoxins, proteins and polyphenols can be found in their composition. Several purification protocols as well as different batteries of assays to prove the biocompatibility of the alginates in vitro have been recently developed. However, little is known about how the use of alginates with different purity grade may affect the host immune response after their implantation in vivo. The present paper investigates the long-term functionality and biocompatibility of murine erythropoietin (EPO) secreting C2C12 cells entrapped in microcapsules elaborated with alginates with different properties (purity, composition and viscosity). Results showed that independently of the alginate type employed, the animals presented elevated hematocrit levels until day 130, remaining at values between 70-87%. However, histological analysis of the explanted devices showed higher overgrowth around non-biomedical grade alginate microcapsules which could be directly related with higher impurity content of this type of alginate. Although EPO delivery may be limited by the formation of a fibrotic layer around non-biomedical grade alginate microcapsules, the high EPO secretion of the encapsulated cells together with the pharmacodynamic behaviour and the angiogenic and immune-modulatory properties of EPO result in no direct correlation between the biocompatibility of the alginate and the therapeutic response obtained. PMID- 17081644 TI - [Does the quality of Web sites related to low back pain meet patients expectations? A systematic review]. AB - INTRODUCTION: An assessment of 36 back-pain-related French-language Web sites, miming a patient search strategy, was previously done by use of evidence-based items. Medical information quality was poor, as already noted about English language Web sites. Thus, patients' expectations may exceed that provided by Web sites with simple medical information. OBJECTIVES: To study whether French language Web sites related to low back pain meet patients expectations and to valid a rating scale including patients' expectations. MATERIALS AND METHOD: First we reviewed French-language Web sites with new keywords and medical gateways. Second, we systematically double assessed back pain-related Web sites with a health care professional and patient-centered scale. RESULTS: We found 30 additional Web sites not found with the previous search, 7 focusing on patient information. The rating scale is valid, and its use on a Web site sample leads to results different from those generated by an evidence-based medicine rating scale but close to a more global assessment. DISCUSSION: French-language Web sites related to low back pain do not meet patients' expectations. Patients participation in Web site assessment or construction could help to close the gap between the expectations of people with low back pain and information delivered by doctors. PMID- 17081645 TI - Brazilian version of the Problem Areas in Diabetes Scale (B-PAID): validation and identification of individuals at high risk for emotional distress. AB - The purpose of this study was to validate the Brazilian version of the Problem Areas in Diabetes Scale (B-PAID) in a sample of outpatients with type 2 diabetes (DM2) and to identify the characteristics of individuals at high risk for emotional distress. A cross-sectional study was conducted in a sample of 146 patients randomly selected among those attending outpatient clinics at a University Hospital in Porto Alegre, Brazil. Information was collected regarding age, gender, known duration of diabetes, type of treatment and level of schooling, and a blood sample was taken to measure HbA1c. B-PAID was compared with the already validated Brazilian versions of Satisfaction with Life (SWL) and WHO Quality of Life-Brief (WHOQOL) Scales. One-hundred forty-six DM2 patients (48% males, 59.7+/-9 years, BMI: 29.7+/-5.2kg/m(2), mean duration: 8.63+/-8 years, HbA(1c): 7.2+/-1.7%) were included. There were significant correlations between B-PAID and SWL (r(p)=0.55, p<0.001) and WHOQOL (r(p)=-0.33, p<0.05) Scales. In a multiple regression analysis the B-PAID Scale was associated with younger age (p=0.006) and lower education level (p=0.05). In conclusion, B-PAID is a reliable, valid outcome measure for Brazilian DM2 patients and identifies patients at increased risk for emotional distress who should be prioritized in psychosocial intervention. PMID- 17081646 TI - A novel index of insulin resistance determined from the homeostasis model assessment index and adiponectin levels in Japanese subjects. AB - Insulin resistance is the principal cause of glucose intolerance and type 2 diabetes and induces progression of severe atherosclerosis in these patients. Adiponectin, the adipose-specific proteins, is known to correlate negatively with insulin resistance in patients with obesity and type 2 diabetes. The purpose of this study was to evaluate the potential of using serum adiponectin levels as a marker of insulin resistance in various states of insulin resistance. Furthermore, we attempted to establish a modified index of the homeostasis model assessment index (HOMA-IR), calculated from the product of serum insulin and plasma glucose levels divided by serum adiponectin levels (HOMA-AD). We recruited 117 Japanese subjects with various degrees of glucose tolerance and determined serum adiponectin levels and insulin sensitivity (M-value) by using the euglycemic hyperinsulinemic clamp technique. M-value, the gold standard index of insulin resistance, correlates significantly and independently with fasting insulin (r=-0.313, P<0.001), glucose (r=-0.319, P<0.001), and adiponectin (r=0.241, P<0.002) levels. M-values were more significantly correlated with HOMA AD (r=-0.643, P<0.001) than HOMA-IR values (r=-0.591, P<0.001). In subjects with moderate hyperglycemia (fasting glucose levels>8.0mmol/L, n=30), HOMA-AD showed a more significant correlation with the M-value than HOMA-IR (r=-0.535, P=0.005 versus r=-0.461, P=0.010). We would therefore like to propose a novel index, HOMA AD, as a simple and adequate index for determining insulin resistance even in diabetic patients with overt hyperglycemia. PMID- 17081647 TI - Application of data mining approaches to drug delivery. AB - Computational approaches play a key role in all areas of the pharmaceutical industry from data mining, experimental and clinical data capture to pharmacoeconomics and adverse events monitoring. They will likely continue to be indispensable assets along with a growing library of software applications. This is primarily due to the increasingly massive amount of biology, chemistry and clinical data, which is now entering the public domain mainly as a result of NIH and commercially funded projects. We are therefore in need of new methods for mining this mountain of data in order to enable new hypothesis generation. The computational approaches include, but are not limited to, database compilation, quantitative structure activity relationships (QSAR), pharmacophores, network visualization models, decision trees, machine learning algorithms and multidimensional data visualization software that could be used to improve drug delivery after mining public and/or proprietary data. We will discuss some areas of unmet needs in the area of data mining for drug delivery that can be addressed with new software tools or databases of relevance to future pharmaceutical projects. PMID- 17081648 TI - Drug transport in corneal epithelium and blood-retina barrier: emerging role of transporters in ocular pharmacokinetics. AB - Corneal epithelium and blood-retina barrier (i.e. retinal capillaries and retinal pigment epithelium (RPE)) are the key membranes that regulate the access of xenobiotics into the ocular tissues. Corneal epithelium limits drug absorption from the lacrimal fluid into the anterior chamber after eyedrop administration, whereas blood-retina barrier restricts the entry of drugs from systemic circulation to the posterior eye segment. Like in general pharmacokinetics, the role of transporters has been considered to be quite limited as compared to the passive diffusion of drugs across the membranes. As the functional role of transporters is being revealed it has become evident that the transporters are widely important in pharmacokinetics. This review updates the current knowledge about the transporters in the corneal epithelium and blood-retina barrier and demonstrates that the information is far from complete. We also show that quite many ocular drugs are known to interact with transporters, but the studies about the expression and function of those transporters in the eye are still sparse. Therefore, the transporters probably have greater role in ocular pharmacokinetics than we currently realise. PMID- 17081649 TI - Hydrogels in controlled release formulations: network design and mathematical modeling. AB - Over the past few decades, advances in hydrogel technologies have spurred development in many biomedical applications including controlled drug delivery. Many novel hydrogel-based delivery matrices have been designed and fabricated to fulfill the ever-increasing needs of the pharmaceutical and medical fields. Mathematical modeling plays an important role in facilitating hydrogel network design by identifying key parameters and molecule release mechanisms. The objective of this article is to review the fundamentals and recent advances in hydrogel network design as well as mathematical modeling approaches related to controlled molecule release from hydrogels. In the first section, the niche roles of hydrogels in controlled release, molecule release mechanisms, and hydrogel design criteria for controlled release applications are discussed. Novel hydrogel systems for drug delivery including biodegradable, smart, and biomimetic hydrogels are reviewed in the second section. Several mechanisms have been elucidated to describe molecule release from polymer hydrogel systems including diffusion, swelling, and chemically-controlled release. The focus of the final part of this article is discussion of emerging hydrogel delivery systems and challenges associated with modeling the performance of these devices. PMID- 17081650 TI - Particulate nanomedicines. PMID- 17081651 TI - The need for bioaugmentation after thermal treatment of a TCE-contaminated aquifer: Laboratory experiments. AB - A microcosm study was conducted to evaluate the need for bioaugmentation after a thermal treatment to anaerobically dechlorinate trichloroethene (TCE) to ethene. The microcosms were either: heated to 100 degrees C and slowly cooled to simulate thermal remediation while bioaugmenting when the declining temperature reached 10 degrees C; or kept at ambient groundwater temperatures (10 degrees C) and bioaugmented for comparison. Aquifer samples from three sediment locations within a TCE-polluted source zone were investigated in duplicate microcosms. In biostimulated (5 mM lactate) and heated microcosms, no conversion of TCE was observed in 4 out of 6 microcosms, and in the remaining microcosms the dechlorination of TCE was incomplete to cDCE (cis-dichloroethene). By comparison, complete TCE dechlorination to ethene was observed in 4 out of 6 heated microcosms that were bioaugmented with a highly enriched dechlorinating mixed culture, KB-1, but no electron donor, and also in 4 of 6 microcosms that were augmented with KB-1 and an electron donor (5 mM lactate). These data suggest that electron donor released during heating, was capable of promoting complete dechlorination coincident with bioaugmentation. Heated microcosms demonstrated less methanogenesis than unheated microcosms, even with elevated H2 concentrations and addition of KB-1, which contains methanogens. This suggests that the heating process suppressed the native microbial community, which can decrease competition with the bioaugmented culture and increase the effectiveness of dechlorination following a thermal treatment. Specifically, cDCE removal rates were four to six times higher in heated than unheated bioaugmented microcosms. This study confirms the need for bioaugmentation following a laboratory thermal treatment to obtain complete dechlorination of TCE. PMID- 17081652 TI - Scaling of spontaneous imbibition data with wettability included. AB - Wettability is a dominant parameter governing spontaneous imbibition. However less attention has been paid to the effect of wettability on the scaling of spontaneous imbibition data. Actually few models can include wettability in scaling of spontaneous imbibition data. To this end, a scaling model has been developed for NAPL (oil)-saturated porous media with different wettability based on the fluid flow mechanisms in porous media. Relative permeability, capillary pressure, initial water saturation, and wettability are considered in the scaling model. Theoretically this scaling model is suitable for both cocurrent and countercurrent spontaneous imbibition. The experimental data of countercurrent spontaneous water imbibition at different wettability cannot be scaled using the frequently used scaling model but can be scaled satisfactorily using the scaling model developed in this study. An analytical solution to the relationship between recovery and imbibition time for linear spontaneous imbibition has also been derived in the case in which gravity is ignored. The analytical solution predicts a linear correlation between the recovery by spontaneous water imbibition and the square root of imbibition time, which has been verified against experimental data. PMID- 17081653 TI - Factors controlling BTEX and chlorinated solvents plume length under natural attenuation conditions. AB - Natural attenuation is presently used at numerous sites where groundwater is contaminated. In order to simulate this attenuation, reactive transport models are often used but they are quite complex and depend on both physical and chemical conditions in the aquifer. As complex numerical models cannot be used to study all possible cases, we develop here analytical solutions to draw general conclusions. Our strategy, called MIKSS (Mixed Instantaneous and Kinetics Superposition Sequence), allows the calculation of the concentrations of all reacting substances in a plume. It is an extension of the superimposition principle that is able to treat the case of joint kinetics and instantaneous reactions. The basic equations have been extended to treat different reactions that occur in the plume core and at its fringe. At first we consider one organic substance degraded under all oxidising conditions (toluene for instance). For this problem the size of the plume depends on the reduced source width and on the ratio of the organic substance concentration to the sum of the electron acceptors' concentrations. For several BTEX substances having different degradation behaviour the formulation is similar, but leads to quite different plume lengths for each substance. Contrary to the case of one substance, the plumes can be quite long and may not satisfy the target risk level. For chlorinated solvents we developed a specific approach to take under consideration all reactions and particularly the competition for hydrogen. A formula is given to assess the size of the plume core, i.e. the zone with highly reducing conditions. The factors influencing the core length are the same as for BTEX (source width, dispersivity, organic carbon content). The size of the TCE plume is calculated from the plume core length and the kinetic constant of TCE degradation. Using assumptions of degradation constants for DCE and VC it is also possible to calculate the longitudinal concentration profile of these substances. The degradation of moderately substituted solvents under oxic conditions reduces the size of their plumes but under these conditions TCE becomes the major threat. Among the conditions studied in this paper, very few chlorinated solvents sites can lead to a negligible risk at an acceptable distance from the source. PMID- 17081654 TI - Identification of differentially expressed genes induced by ozone stress in sensitive and tolerant poplar hybrids. AB - The differential expression of genes induced by an acute ozone treatment was analysed in two poplar clones, i.e. Populus deltoides x maximowiczii, Eridano clone, and Populus x euoramericana, I-214 clone, respectively, sensitive and tolerant to this pollutant, performing suppression subtractive hybridization (SSH). From the obtained cDNA libraries several clones were obtained, which corresponded to differentially regulated genes. Preliminary expression analyses of four genes, Fs23A-LRP, Ft33B-CaBP, Ft312B-WRKY, and Ft32C-WAK identified by the primary screening, were conducted by semi-quantitative RT-PCR to evaluate the ozone responsiveness of the libraries. The most interesting finding is the co activation of a wall associated kinase and a WRKY transcription factor in response to O(3) stress. PMID- 17081655 TI - Vagus nerve stimulation acutely alters food craving in adults with depression. AB - Vagus nerve stimulation (VNS) is now available as a treatment for epilepsy and treatment-resistant depression. The vagus nerve plays a central role in satiety and short-term regulation of food intake and research suggests a relationship between VNS and weight loss. The underlying mechanisms of this relationship are unknown. The purpose of the current study was to determine whether acute cervical VNS might temporarily alter food cravings. Thirty-three participants were recruited for three groups; depression VNS, depression non-VNS, and healthy controls. Participants viewed 22 computerized images of foods twice in one session and completed ratings for food cravings after each image. The VNS participants' devices were turned on for one viewing of an image and off for the other (randomized order). Participants were blind to VNS condition (on versus off). Acute VNS device activation was associated with a significant change in cravings-ratings for sweet foods. A significant proportion of variability in VNS related changes in cravings was accounted for by patients' clinical VNS device settings, acute level of depression, and body mass. Further studies are warranted addressing how acute or chronic VNS might modify eating behavior and weight. PMID- 17081656 TI - Frequent feeding delays the gastric emptying of a subsequent meal. AB - To assess the suitability of the 13C-octanoic acid breath test for measuring gastric emptying in circumstances other than the post-absorptive state, a preliminary study was performed where 6 hourly spaced isoenergetic meals preceded the determination of gastric emptying of a subsequent 2 MJ meal. Emptying was measured in three individuals on four separate occasions, with a reproducibility of 8%. A crossover study was then conducted to test the hypothesis that meal frequency can modulate the gastric emptying of a subsequent meal, with the potential to influence appetite regulation. Sixteen subjects were fed to energy balance, receiving food either as 2 isoenergetic meals 3 h apart or 6 isoenergetic meals fed hourly. Gastric emptying of a subsequent 2 MJ meal was investigated. Visual analogue scales were used throughout to assess appetite. The maximum rate of gastric emptying was unchanged but the onset of emptying was delayed by the more frequent feeding pattern. There was no significant difference in subjective appetite before or after the test meal. In conclusion, short-term increases in feeding frequency delayed the gastric emptying of a subsequent meal, but significant effects on post-meal appetite could not be demonstrated. PMID- 17081657 TI - Psychological distress, personality, and adjustment among nursing students. AB - Psychological distress and poor adjustment among a significant number of nursing students is an important issue facing nursing education. The concerns need to be studied in detail and solutions need to be built into the nursing course in order to help students with such difficulty. This study used a cross-sectional survey design to study psychological distress, personality and adjustment among nursing students attending the College of Nursing, Christian Medical College, Vellore, India. One hundred and forty five nursing students were assessed using the General Health Questionnaire 12, the Eysenck Personality Questionnaire, and the Bell's Adjustment Inventory to investigate psychological distress, personality profile and adjustment, respectively. Thirty participants (20.7%) of the 145 students assessed reported high scores on the General Health Questionnaire. Psychological distress was significantly associated with having neurotic personality and adjustment difficulties in different areas of functioning. PMID- 17081658 TI - Implementing reflection: insights from pre-registration mental health students. AB - Reflection and reflective practice continues to be contentious issues in nursing. The focus of this article is the use of reflection by pre-registration mental health students. The broad aim of this preliminary study was to discover student mental health nurses' perceptions of reflection as a learning strategy during clinical placement. Using a constructivist grounded theory methodology [Charmaz, K., 2000. Grounded theory: Objectivist and Constructivist Methods. In: Denzin, N., Lincoln, Y. (Eds.), Handbook of Qualitative Research, second ed. Sage, Thousand Oaks, California], five students were interviewed individually in their clinical placements. Data analysis revealed three major categories: understanding the process of reflection, using reflection in clinical practice, and needing support and guidance. Findings indicated that students were primarily using reflection-on-action, but to varying extents. Overall, students felt that reflection facilitated their learning. Factors were discovered that both helped and hindered students' use of reflection. These included level of preparation to reflect, a limited culture of reflection and the level of support from preceptors, clinical staff, clinical placement co-ordinators, and lecturers. In conclusion, it appears that a collaborative approach between students, Health Service Providers and institutes of nursing is vital for the successful development and implementation of reflective learning strategies in clinical placement. Suggestions are made as to how a collaborative approach may be developed to enhance this process. PMID- 17081659 TI - The value of an evidence based practice module to skill development. AB - BACKGROUND: A degree level EBP module is undertaken by students on post qualifying programmes to develop critical appraisal skills gained at diploma level. Despite training in EBP skills, there is some suggestion that health care professionals lack confidence in this area and fail to make the links between evidence and practice. The purpose of this study was to investigate this issue further. AIMS: To identify how the EBP skills developed on the module were utilised in practice, and to identify what local barriers there were in using these skills. METHODS: A survey of all students who had completed the module within the past three years (n=393) followed by interviews to explore further issues that emerged from the survey. RESULTS: One hundred and ninety one completed questionnaires (48.6% response rate). The results showed that the students perceived themselves as 'quite skilled' or 'competent' with regard to the key EBP skills, and that completion of the module had contributed 'greatly' to this skill acquisition. Results from the interviews showed that time and cultural issues were the greatest barriers to using and developing further the EBP skills gained. CONCLUSIONS: The value of the module was apparent. For skills to be developed further in the work place, cultural changes need to occur. PMID- 17081660 TI - Development of lactococcal GEM-based pneumococcal vaccines. AB - We report the development of a novel protein-based nasal vaccine against Streptococcus pneumoniae, in which three pneumococcal proteins were displayed on the surface of a non-recombinant, killed Lactococcus lactis-derived delivery system, called Gram-positive Enhancer Matrix (GEM). The GEM particles induced the production of the proinflammatory cytokine tumour necrosis factor-alpha (TNF alpha) by macrophages as well as the maturation of dendritic cells. The pneumococcal proteins IgA1 protease (IgA1p), putative proteinase maturation protein A (PpmA) and streptococcal lipoprotein A (SlrA) were anchored in trans to the surface of the GEM particles after recombinant production of the antigens in L. lactis as hybrids with a lactococcal cell wall binding domain, named Protein Anchor domain (PA). Intranasal immunisation with the SlrA-IgA1p or trivalent vaccine combinations without additional adjuvants showed significant protection against fatal pneumococcal pneumonia in mice. The GEM-based trivalent vaccine is a potential pneumococcal vaccine candidate that is expected to be easy to administer, safe and affordable to produce. PMID- 17081661 TI - AAV vectors encoding malarial antigens stimulate antigen-specific immunity but do not protect from parasite infection. AB - This study explores the utility of recombinant adeno-associated virus (rAAV) as a genetic vaccine delivery system using muscle as a target tissue. A single injection of rAAV encoding the malarial antigens MSP4 (Plasmodium falciparum) or MSP4/5 (Plasmodium yoelii) stimulated long-term antigen-specific antibody responses. Anti-MSP4/5 immunity stimulated by AAV was not protective against P. yoelii infection and efforts taken to augment antibody responses against MSP4/5, either by priming with plasmid DNA or AAV and boosting with rAAV were unsuccessful. Alternative strategies such as inclusion of genetic adjuvants into the AAV vector will be necessary to stimulate an adequate level of anti-malarial protective immunity in this model. PMID- 17081662 TI - Estimating vaccine effectiveness against laboratory-confirmed influenza using a sentinel physician network: results from the 2005-2006 season of dual A and B vaccine mismatch in Canada. AB - INTRODUCTION: We report a case-control design using a sentinel physician network to estimate vaccine effectiveness (VE) against laboratory-confirmed, medically attended influenza (LC-MAI) and provide results for the 2005-2006 season of dual A and B vaccine mismatch in Canada. METHODS: Participants were patients >or=5 years of age presenting with influenza-like illness (ILI) to a sentinel physician in British Columbia, Canada between November 1, 2005 and April 30, 2006. Cases were participants in whom influenza was identified; controls tested negative for influenza A and B by PCR, R-mix and culture. Isolates were characterized by gene sequencing and hemagglutination-inhibition (HI) assays. Odds ratios (OR) for LC MAI in vaccinated versus non-vaccinated persons were derived with adjustment for age and chronic conditions. VE was estimated as [1-OR (vaccinated/unvaccinated)]. RESULTS: The sample included 442 patient visits: median age was 26 years, 10% were >or=65 years, 15% had a chronic condition and 22% received the 2005-2006 trivalent inactivated influenza vaccine >or=2 weeks before ILI onset. Two hundred and six participants were positive for influenza; 107 (52%) had influenza A/H3N2 and 99 (48%) had influenza B/Victoria lineage. Gene sequencing identified mutations away from the vaccine strain at key antigenic binding sites of the hemagglutinin (HA) protein of H3N2 isolates; the neuraminidase (NA) protein was conserved. Based on HI assays, three-quarters of influenza A and all B isolates were mismatched to the 2005-2006 vaccine. Point estimates for VE against LC-MAI were in the range of 50 to 70% for both types of influenza. CONCLUSION: 2005-2006 was the third consecutive season of vaccine mismatch based on varying HA for the A/H3N2 component and the third also for the B component since 2001. Vaccine mismatch resulted in diminished VE but substantial cross-protection. More timely detection of drift variants through gene sequencing of isolates facilitates interpretation of VE results. Since it may be more antigenically conserved, the vaccine content and contribution of NA to overall VE should be further evaluated for both A and B components. Infrastructure for real-time epidemiologic assessment of vaccine performance is important annually and in preparation for a pandemic. PMID- 17081663 TI - Predicting the transfer of 137Cs to rice plants by a dynamic compartment model with a consideration of the soil properties. AB - This paper describes the predictions of the transfer of 137Cs to rice plants following soil deposition during a non-growth season of rice by a dynamic compartment model considering the soil properties, and their comparison with experimental results. Major processes considered in the model were percolation, soil mixing by plowing, plant uptake, leaching to deep soil, fixation to clay mineral, and time-dependent growth of a plant. To consider the effects of the soil properties (pH, clay mineral, organic matter content, and exchangeable K) on plant uptake and the leaching rates of 137Cs in a root zone soil, the Absalom model was used for the present model. The 137Cs aggregated transfer factors (TFa, m2kg-1 dry plant) of rice plants for two consecutive cultivation years were obtained as a result of simulated 137Cs soil deposition experiments with 17 paddy soils of different properties, all of which were performed before transplanting of the rice. Observed 137Cs TFa values of the rice plants did not show an evident trend for the pH and clay content of the soil properties, while they increased with an increasing organic matter content and a decreasing exchangeable K concentration. Predicted 137Cs TFa values of the rice plants were found to be comparable with those observed. PMID- 17081664 TI - Dynamic periosteal elevation. AB - The osteoinductive potential of the periosteum can be stimulated by raising the periosteum using a type of distraction. This was tested and confirmed in an animal experiment in 6 Goettingen minipigs. A titanium mesh was implanted beneath the periosteum and then raised. Bone formed underneath the mesh, with rows of micro-pillars similar to those found after osteodistraction. The main advantages of dynamic periosteal elevation are that invasion and morbidity are minimal. Clinically it might be applicable in craniomaxillofacial surgery, in augmentation before implantation, and in reconstruction of the skull. PMID- 17081665 TI - Use of methylene blue as a diagnostic aid in early detection of oral cancer and precancerous lesions. AB - We assessed the sensitivity, specificity, and positive and negative predictive value of methylene blue staining in the diagnosis of oral cancer in 58 patients. The sensitivity was 90%, the specificity 69%, positive predictive value 74%, and negative predictive value 87%. Because of the number of false negatives and false positives we recommend that the diagnosis should always be confirmed by histopathological examination of a biopsy specimen. Methylene blue staining may, however, be useful as a screening tool for oral cancer in large, high-risk groups in a similar way to the more expensive toluidine blue. PMID- 17081666 TI - Nanomedicine: An unresolved regulatory issue. AB - Nanomedicine is a science that uses nanotechnology to maintain and improve human health at the molecular scale. Current and potential applications of nanotechnology in medicine range from research involving diagnostic devices, drug delivery vehicles to enhanced gene therapy and tissue engineering procedures. Its advantage over conventional medicine lies on its size. Particle size has effect on serum lifetime and pattern of deposition. This allows drugs of nanosize be used in lower concentration and has an earlier onset of therapeutic action. It also provides materials for controlled drug delivery by directing carriers to a specific location. Major efforts are underway, however, very little attention is devoted to assessment of health risks to human or to the ecosystem. Inhaled nanoparticles have already been related to lung injury. It is recognized that physico-chemical properties in conjunction with environmental factors and stability of the nanomaterial all contribute to the overall toxicological responses. Nanotoxicological information, currently insufficient, will be vital in aiding academia, industry and regulatory bodies in elucidating the mechanisms of action, balancing its risk and benefit, thus maximizing the utility of these materials in medicine without compromising public health and environmental integrity. PMID- 17081668 TI - Localised reductions in gyrification in the posterior cingulate: schizophrenia and controls. AB - Several authors report changes in the volume and degree of cortical folding in several presumptive regions of schizophrenia patients. However, no research, to our knowledge, has looked for schizophrenia related differences in cortical volume and degree of cortical folding in post-mortem posterior cingulate cortex (PCCx). METHOD: Brain tissues from 9 people who suffered from schizophrenia (DSM IV) and 9 controls were cut into 3 mm coronal slices. Three alternative PCCx blocks were available for research. The 3 PCCx blocks were cut into 50 microm sections. The volume and gyrification index (GI) were measured in 15 tissue sections per brain (5 sections per block). RESULTS: People who suffered with schizophrenia showed significant reductions in GI in rostral PCCx, trend reductions were seen in medial and caudal PCCx. In addition, the average volume of the rostral tissue sections was significantly lower in the schizophrenia cohort, suggesting that schizophrenia is associated with reduced volume in the rostral PCCx. However, a true volumetric assessment of the whole PCCx, rather than a limited number of sections from three alternative blocks, is needed to confirm such a hypothesis. PMID- 17081667 TI - Genetic counseling communication with an African American BRCA1 kindred. AB - We studied communication in genetic counseling sessions conducted with an African American, Breast Cancer 1, Early Onset (BRCA1) kindred in the USA. The Roter Interaction Analysis System (RIAS) was used to code and compare two sessions of 46 participants (26 females and 20 males) before and after they underwent genetic testing. Three certified genetic counselors and one medical geneticist conducted the sessions. When compared to pre-test communication, most of the providers' post-test communication was devoted to the provision of biomedical information (including screening recommendations) with fewer questions and psychosocial statements. Clients contributed a similar proportion to the total session dialogue in pre- and post-test sessions (40%). A larger proportion of their post test session was devoted to indicating receptiveness to provider information than in the pre-test session. We found when providers were informing clients that they were BRCA1 mutation carriers, they provided more biomedical and psychosocial information and asked more psychosocial questions than when talking with non carriers. This study provides the first description of genetic counseling communication for pre- and post-test BRCA1 sessions with African American individuals. PMID- 17081669 TI - Quetiapine regulates the stress-induced increase in corticotropin-releasing factor mRNA expression in the rat hypothalamus. AB - Corticotropin-releasing factor (CRF) is a key regulator of the stress response. We investigated the effects of the atypical antipsychotic drug quetiapine on CRF mRNA expression in the rat hypothalamus following immobilization stress. Pretreatment with 10 mg/kg quetiapine significantly reduced the immobilization stress-induced increase in CRF mRNA expression in the paraventricular nucleus of the hypothalamus. These results suggest that quetiapine may modulate the stress response via regulation of CRF mRNA expression. PMID- 17081670 TI - Glial cell activation in a subgroup of patients with schizophrenia indicated by increased S100B serum concentrations and elevated myo-inositol. AB - Post-mortem and in-vivo studies support the hypothesis that astrocytes might be involved in the pathogenesis of schizophrenia. To further substantiate this hypothesis two markers of astroglial activation (myo-inositol, S100B) acquired with independent methods ((1)H-MRS, quantitative immunoassay) were concomitantly measured in schizophrenic patients. Patients with increased S100B levels showed elevated myo-inositol concentrations. This pilot study demonstrates a concomitant elevation of two markers indicating astrocyte activation in a subgroup of schizophrenic patients. PMID- 17081671 TI - Evaluation of the safety and toxicity of the oligomerized polyphenol Oligonol. AB - Oligonol((R)) is an optimised phenolic product containing catechin-type monomers and lower oligomers of proanthocyanidin that emanate from a technology process which converts polyphenol polymers into oligomers. In a single dose toxicity study administration of Oligonol (2000mg/kg bw) by gavage for 4 weeks was found to be safe with no side effects (such as abnormal behavior and alopecia). Body weight gain and food consumption were within normal range. Oligonol had no observed toxicity at the dose (1/25 of LD(50)) administered for 6 months. This suggests that Oligonol is safe at repeated human intakes of Oligonol in doses lower than 200mg/day. The highest dose used in this study is equal to 12g daily for an adult man with 60kg body weight. The LD(50) was calculated to be 5.0g/kg body weight (95% confidence limit: 3.5-6.4g/kg). Studies conducted on 30 healthy volunteers consuming Oligonol at doses of 100mg/day and 200mg/day for 92 days showed good bioavailability. The biochemical parameters attesting to liver and kidney functions as well as the hematological parameters were within the normal ranges. The potential of Oligonol to induce gene mutation (a reverse mutation test) was tested using Salmonella typhimurium TA98, TA100, TA104, TA1535, TA153 and Escherichia coli WP2uvrA. Oligonol was not mutagenic to the tester strains. The lack of toxicity supports the potential use of Oligonol as a food or dietary supplement and for use as an additive in pharmaceutical and cosmetological applications. PMID- 17081672 TI - Toward understanding respiratory sinus arrhythmia: relations to cardiac vagal tone, evolution and biobehavioral functions. AB - Respiratory sinus arrhythmia (RSA, or high-frequency heart-rate variability) is frequently employed as an index of cardiac vagal tone or even believed to be a direct measure of vagal tone. However, there are many significant caveats regarding vagal tone interpretation: 1. Respiratory parameters can confound relations between RSA and cardiac vagal tone.2. Although intraindividual relations between RSA and cardiac vagal control are often strong, interindividual associations may be modest.3. RSA measurement is profoundly influenced by concurrent levels of momentary physical activity, which can bias estimation of individual differences in vagal tone.4. RSA magnitude is affected by beta adrenergic tone.5. RSA and cardiac vagal tone can dissociate under certain circumstances.6. The polyvagal theory contains evolution-based speculations that relate RSA, vagal tone and behavioral phenomena. We present evidence that the polyvagal theory does not accurately depict evolution of vagal control of heart rate variability, and that it ignores the phenomenon of cardiac aliasing and disregards the evolution of a functional role for vagal control of the heart, from cardiorespiratory synchrony in fish to RSA in mammals. Unawareness of these issues can lead to misinterpretation of cardiovascular autonomic mechanisms. On the other hand, RSA has been shown to often provide a reasonable reflection of cardiac vagal tone when the above-mentioned complexities are considered. Finally, a recent hypothesis is expanded upon, in which RSA plays a primary role in regulation of energy exchange by means of synchronizing respiratory and cardiovascular processes during metabolic and behavioral change. PMID- 17081673 TI - Engineered internal noise stochastic resonator in gene network: a model study. AB - Based on a genetic bistable switch model coupled with a gene oscillator model, we have constructed a mesoscopic stochastic model for the coupled synthetic gene network, and studied how internal noise would influence the oscillation of such a system. We found that the state-to-state transitions can occur if the internal noise is taken into account, and the performance of resulting oscillation can reach a maximum in a certain internal noise level, which indicates the occurrence of internal noise stochastic resonance (SR) and makes the coupled gene network work as a stochastic resonator. The potential role of such an effect on gene expression systems is also discussed. PMID- 17081674 TI - Penile prosthesis implantation: size matters. PMID- 17081675 TI - Effect of sildenafil and rolipram on adrenergic responses in isolated human and monkey corpus cavernosum. AB - OBJECTIVE: Evaluate and compare effects of phosphodiesterase inhibitors (PDEIs), sildenafil and rolipram, on adrenergic contractile responses of human and monkey cavernosal smooth muscle. METHODS: Human penises were obtained from patients undergoing gender reassignment surgery. Isolated human and monkey corpus cavernosum (CC) strips were suspended in tissue bath chambers for isometric tension experiments. The effects of the drugs on precontracted monkey and human CC and neurogenic contractions in human CC were investigated. RESULTS: Both sildenafil and rolipram induced concentration-dependent relaxations of human and monkey CC strips precontracted with noradrenaline (NA). The IC(50) values, determined by reverse regression for nitroglycerin (NTG), isoprenaline, and sildenafil in monkey CC, were, respectively, 1.5+/-0.9x10(-7) M, 3.7+/-0.6x10(-6) M, and 1.7+/-0.7x10(-5) M. Similarly, in human CC muscle, sildenafil was weaker than NTG as a muscle relaxant. Sildenafil, 1.5 microM, reduced neurogenic contractions in human CC due to stimulation of predominantly adrenergic nerves. The suppressant effect of sildenafil on adrenergic transmission was attenuated in CC strips pretreated with N omega-nitro-L-arginine and overcome with a higher stimulus frequency or tetraethylammonium. Rolipram partially inhibited adrenergic excitatory response but without significantly affecting NA-induced contraction. CONCLUSIONS: Sildenafil and rolipram induced concentration-dependent reversal of human and monkey CC tone mediated by NA. Both PDEIs attenuated contractile adrenergic response of human CC to electrical stimulation. The results also underline the importance of the cyclic adenosine monophosphate-dependent signalling pathway in regulating the tone. PDE4 inhibition in CC is an additional mechanism for erection and potential therapeutic adjunct. PMID- 17081676 TI - Patient characteristics associated with quality of life in European women seeking treatment for urinary incontinence: results from PURE. AB - OBJECTIVE: To investigate the association between patient characteristics and disease-specific and generic quality of life (QOL) as well as the degree of bother in women seeking treatment for urinary incontinence (UI). METHODS: The Prospective Urinary Incontinence Research (PURE) was a 6-mo observational study with 1055 physicians from 15 European countries enrolling 9487 women. QOL was assessed at the enrolment visit using the urinary Incontinence Quality of Life questionnaire (I-QOL) and the generic EQ-5D. A single-item instrument was used to measure the degree of bother. UI severity was assessed using the Sandvik Index. UI was categorised into stress (SUI), mixed (MUI), and urge (UUI) urinary incontinence by a patient-administered instrument (Stress and Urge Incontinence Questionnaire [S/UIQ]). Multivariate linear (I-QOL, EQ-5D Visual Analogue Scale) and logistic (bother, EQ-5D health state index) regressions were performed. RESULTS: Mean total I-QOL scores were significantly and independently associated with UI severity, nocturia, age, UI subtype, number of selected concomitant medical conditions, length of suffering from UI before contacting a doctor, smoking status, ongoing use of UI medication, and country. After adjusting for all the covariates, the total I-QOL scores for SUI, MUI, and UUI were 62.7, 53.8 and 60.1, respectively. As with I-QOL, UI severity was also the most important predictor for bother. The number of concomitant medical conditions, together with UI severity, was the variable most strongly associated with EQ-5D. CONCLUSION: In addition to the UI subtypes, severity of UI should be given more importance in treatment algorithms and in treatment decision-making by both the patient and the physician. PMID- 17081677 TI - The pattern and progression of lower urinary tract symptoms after transurethral prostatectomy compared with those seen in the general population. AB - OBJECTIVES: Men undergoing transurethral prostatectomy (TURP) often have some level of residual symptoms after this operation. Reliance on symptom scores to assess outcomes means there is relatively little information on the exact pattern of such symptoms. The aim of this study was to describe the pattern, progression, and associated bother of lower urinary tract symptoms after TURP. METHODS: Detailed information on urinary symptoms, prostatectomy status, and associated bother was obtained from a randomly selected sample of community-dwelling men by using a postal questionnaire. Rates of individual urinary symptoms in the postprostatectomy population were calculated, and compared with those in the background population and those in secondary care. RESULTS: Of 7640 men who responded, 422 had undergone prostatectomy. Symptom levels remained significantly higher in the postoperative group than the background population, with 19.4% experiencing four or more symptoms on a weekly basis. The pattern of symptoms was similar to that seen in the general population. Men in the post-TURP group experienced greater levels of bother when symptom level (odds ratio: 2.69; 95%CI, 1.80-4.01) was controlled. Both symptom and bother, however, were significantly lower than those in people referred for secondary care treatment of their symptoms. CONCLUSIONS: Men with a previous prostatectomy still report high levels of urinary symptoms and appear to experience greater levels of bother from these symptoms than men in the general population. PMID- 17081678 TI - Chemotherapy in patients with teratoma with malignant transformation. AB - OBJECTIVE: Germ-cell tumours (GCTs) with a non-GCT malignant component are a unique and rare phenomenon called teratoma with malignant transformation (TMT). The only published series of patients with TMT treated with chemotherapy comprised 10 patients. We report here our experience in treating 14 patients with TMT. PATIENTS AND METHODS: Sarcoma was identified in 10 of 14 patients, with rhabdomyosarcoma ranking first (n=4). Other histological types included adenocarcinoma (n=3) and bronchoalveolar carcinoma (n=1). Immunohistochemistry was performed to help in identifying the malignant non-GCT component. RESULTS: Primary treatment consisted of surgery alone in 4 patients. The remaining 10 patients received first-line cisplatin-based chemotherapy with resection of residual masses (n=5): 4 patients had a complete response and 5 had a partial response. Overall, 9 patients developed a relapse with a median time of 84 mo (range: 6-168). At relapse, 8 patients received a chemotherapy regimen directed to the non-GCT component. Four of these patients achieved a partial response. With a median follow-up of 59 mo (range: 3-180), 4 of 14 patients are alive, including 3 who are disease-free. CONCLUSION: To our knowledge, this is by far the largest reported European series of chemotherapy in TMT. Although TMT has a poor prognosis compared to GCT, its management may be improved by adapted chemotherapy associated with surgical resection of residual masses. PMID- 17081679 TI - Image-guided biopsy-diagnosed renal cell carcinoma: critical appraisal of technique and long-term follow-up. AB - OBJECTIVES: To critically appraise and determine the impact of image-guided biopsy on the management of indeterminate renal masses. A comparison of long-term follow-up of renal cell carcinoma (RCC) diagnosed by image-guided biopsy and radiologically obvious RCC was also carried out. PATIENTS AND METHODS: Data were collected for all the consecutive patients requiring renal core biopsies for the diagnosis of indeterminate renal masses between January 1996 and January 2006. The long-term outcome of diagnostic and nondiagnostic renal biopsies was assessed. Furthermore, the long-term outcome of RCC diagnosed following biopsies was compared with nonbiopsy radical nephrectomy done during the same time period. RESULTS: Of the 70 biopsy procedures performed, 9 were nondiagnostic and 61 were diagnostic on histopathologic examinations (17 benign and 44 malignant). The histopathology of all radical nephrectomies was identical to the pathology of biopsy specimens. Of the nine nondiagnostic cases, one patient had a repeat biopsy that was confirmed as RCC. Six patients including the case diagnosed to have RCC on repeat biopsy underwent radical nephrectomy in the nondiagnostic group. The histopathology revealed RCC in four, and angiomyelolipoma and pyelonephritis in one each. The remaining three nondiagnostic cases are under follow-up; there has been no change in the size of the lesions in a mean follow up of 32 mo (range: 12-52). There has been no change in the size of benign lesions at a mean follow-up of 29 mo (range: 3-72). The procedure-related complication in the form of bleeding following biopsy was observed in one patient, which settled conservatively. There was no statistically significant difference (chi-square=1.134 and p value equal to 0.379) in the recurrence rate and metastases between the biopsy radical nephrectomy and nonbiopsy radical nephrectomy groups for the same stage of disease during the same period. CONCLUSIONS: Image-guided biopsy is safe and accurately characterises indeterminate renal masses. A repeat biopsy protocol is useful in case of a nondiagnostic first biopsy. The long-term outcome following radical nephrectomy of biopsy-diagnosed RCC does not differ from the radiologically obvious RCC. PMID- 17081680 TI - Real time computation: zooming in on population codes. AB - Information processing in nervous systems intricately combines computation at the neuronal and network levels. Many computations may be envisioned as sequences of signal processing steps along some pathway. How can information encoded by single cells be mapped onto network population codes, and how do different modules or layers in the computation synchronize their communication and computation? These fundamental questions are particularly severe when dealing with real time streams of inputs. Here we study this problem within the context of a minimal signal perception task. In particular, we encode neuronal information by externally applying a space- and time-localized stimulus to individual neurons within a network. We show that a pulse-coupled recurrent neural network can successfully handle this task in real time, and obeys three key requirements: (i) stimulus dependence, (ii) initial-conditions independence, and (iii) accessibility by a readout mechanism. In particular, we suggest that the network's overall level of activity can be used as a temporal cue for a robust readout mechanism. Within this framework, the network can rapidly map a local stimulus onto a population code that can then be reliably read out during some narrow but well defined window of time. PMID- 17081681 TI - Determination of hyperglycosylated human chorionic gonadotropin produced by malignant gestational trophoblastic neoplasias and male germ cell tumors using a lectin-based immunoassay and surface plasmon resonance. AB - The ability to reliably detect aberrant glycosylation of human chorionic gonadotropin (hCG) may have profound implications for the diagnosis and monitoring of malignant gestational trophoblastic neoplasia, germ cell tumors, other malignancies, and pregnancy complications. To become a clinically useful assay, however, this discrimination of glycoforms should be possible on minimally treated biological specimens. Towards this end, we have developed a lectin-based sandwich-type immunoassay to compare the glycosylation patterns of hCG among urine specimens from patients presenting with a normal pregnancy, invasive mole, choriocarcinoma, and male germ cell tumors using carbohydrate-free antibody fragments as capture reagents and a panel of eight lectins, five recognizing neutral sugars and three recognizing sialic acid. There was no significant difference in the binding of any of the lectins to hCG in the urine of women over the gestational range of 6-38 weeks. Three lectins, however, exhibited differential binding to urinary hCG derived from these normal pregnant controls and that from patients with malignant forms of gestational trophoblastic disease and male germ cell tumors. Galanthus nivalis agglutinin and Maackia amurensis lectin, which bind terminal mannose and alpha(2-3)sialic acid, respectively, preferentially bound pregnancy-derived hCG, whereas the lectin, wheat germ agglutinin, which binds sialic acid and beta(1-4)N-acetylglucosamine, exhibited decreased binding to pregnancy-derived hCG compared to that from patients with male germ cell tumors and malignant gestational trophoblastic neoplasia. The differential binding observed with these three promising lectins is most encouraging and warrants further examination. The experimental paradigm also holds promise for the development of comparable assays for other glycosylated tumor markers. PMID- 17081682 TI - The tale of follitropin receptor diversity: a recipe for fine tuning gonadal responses? AB - The original concept (dogma) of a single FSH receptor entity coupling to G(s) protein to activate adenylate cyclase and producing cAMP as second messenger appears inadequate to explain pleiotropic actions of the hormone. The identification and expression of alternatively spliced gonadotropin receptors, suggest that alternative splicing could serve as a mechanism for creating receptor diversity. Studies focused on sheep and mouse gonadal tissues show that the single large gene of approximately 250kb is a modular structure whose pre mRNA undergoes alternative splicing creating several subtypes (at least four FSH R1 to R4 identified to date). With segments of the N-terminus that are identical different topographies are generated by differing carboxyl termini. The same gene thus produces receptor types with different motifs that can display dominant positive, dominant negative, growth factor/cytokine type and potentially soluble binding protein features. Functional relevance is shown by modulation of receptor variants during hormonal stimulation. Presence of equivalent segments of the gene in the human and bovine suggests conservation and predicts similarity in structures and function. Thus, the complex cellular biology of follitropin receptors that may interact differently with polymorphic forms (glycosylation variants) of FSH represents an intricate scheme to regulate hormone signaling. PMID- 17081683 TI - Beneficial effects of prolactin and laminin on human pancreatic islet-cell cultures. AB - The problem of pancreas donor shortage could be addressed through in vitro islet cell proliferation prior to transplantation into diabetic patients. Therefore, we set out to evaluate the effects of prolactin (rhPRL) and laminin on primary cultures of human pancreatic islets. Our results showed that rhPRL induced an increase in islet-cell number and in cumulative insulin secretion (p<0.01). However, glucose-induced insulin secretion was enhanced only in the presence of both laminin and rhPRL. In addition, we describe, for the first time in human islets, the PRL-induced activation of JAK2, and signal transducer and activator of transcription (STAT) 1, 3 and 5. Our results demonstrate a significant beneficial effect of rhPRL and laminin on human islets and support widely held notion that the closer physiological stimuli and environment of beta cells are mimicked, the better are the results in cell proliferation and secretory function, both essential for successful islet transplantation. PMID- 17081684 TI - Hyperglycosylated hCG: a variant with separate biological functions to regular hCG. AB - Hyperglycosylated hCG (hCG-H) is an over-glycosylated variant of hCG. While regular hCG is produced by differentiated syncytotrophoblast cells, hCG-H is independently secreted by stem cytotrophoblast cells. hCG-H has an independent function to regular hCG. It is the direct promoter of trophoblast invasion and malignancy. Invasion as in implantation of pregnancy and malignancy as in choriocarcinoma. Neither will occur in the absence of hCG-H. hCG-H measurements have multiple functions. Primarily or exclusively hCG-H is produced at the time of implantation of pregnancy and in the 2 weeks that follows. As such, a good pregnancy test should measure regular hCG and hCG-H equally. This is not commonly the case. Most tests poorly detect hCG-H. New pregnancy tests are needed, including those that measure only hCG-H. Considering that hCG-H is critical for implantation, hCG-H is also invaluable for determining pregnancy outcome and detecting failures. It makes a considerable more accurate test for detecting pregnancy failures and ectopic pregnancies than regular hCG. Down syndrome pregnancies are marked by poor trophoblast differentiation. As such, they are very well marked by using a combination of hCG-H measurements and other screening tests. hCG-H is also an absolute tumor marker for malignant or invasive gestational trophoblastic disease, it can discriminate active and inactive (quiescent) disease, and the need for chemotherapy. PMID- 17081685 TI - Mitochondrial DNA G10398A polymorphism imparts maternal Haplogroup N a risk for breast and esophageal cancer. AB - Mitochondria are the major source of Reactive Oxygen Species (ROS) and mtDNA G10398A (Ala-->Thr) polymorphism, proposed to be involved in increased ROS production, has been shown in association with invasive breast cancer in African American (AA) women [J.A. Canter, A.R. Kallianpur, F.F. Parl, R.C. Millikan, Mitochondrial DNA G10398A polymorphism and invasive breast cancer in African American women, Cancer Res. 65 (2005) 8028-8033] and prostate cancer in AA men [M.P. Mims, T.G. Hayes, S. Zheng, S.M. Leal, A. Frolov, M.M. Ittmann, et al., Mitochondrial DNA G10398A polymorphism and invasive breast cancer in African American women, Cancer Res. 66 (2006) 1880; author reply 1880-1881]. The role of mitochondria, however, in cancer development has been in question recently [A. Salas, Y.G. Yao, V. Macaulay, A. Vega, A. Carracedo, H.J. Bandelt, A critical reassessment of the role of mitochondria in tumorigenesis, PLoS Med. 2 (2005) e296], which has made it pertinent to analyze the data and test the hypotheses by conducting fresh case-control studies. This study, therefore, makes an attempt to validate the exclusive presence of mtG10398A (Ala-->Thr) polymorphism in a haplotype constituting mtDNA haplogroup N and its sublineages, imparting this group a higher risk for breast cancer, based on the re-analyses of approximately 1000 complete human mtDNA sequences worldwide and collated information on 2334 individuals belonging to 18 regions in India. The conclusion drawn of mt10398A allele providing a risk towards cancer is confirmed in a case-control comparison study of 124 sporadic breast cancer patients and 273 controls; and 55 squamous cell carcinoma of esophagus, ESCC, and 163 controls, matched for age, ethnicity and sex from north India. It is further apparent from the study that such a mtDNA polymorphism background provides a higher risk for the cancers of the tissues which could be affected by environmental insults directly as in the ESCC, observed with a high acquired (somatic) rate of mutation in p53 when compared to the breast cancer, suggesting that the mtDNA variants that arose as energetic adaptations, influence our health differentially under different environment conditions and a given genetic background of the mt genome. PMID- 17081686 TI - Functional characterization of human MutY homolog (hMYH) missense mutation (R231L) that is linked with hMYH-associated polyposis. AB - The MutY homolog (MYH) can excise adenines misincorporated opposite to guanines or 7,8-dihydro-8-oxo-guanines (8-oxoG) during DNA replication; thereby preventing G:C to T:A transversions. Germline mutations in the human MYH gene are associated with recessive inheritance of colorectal adenomatous polyposis (MAP). Here, we characterize one newly identified MAP-associated MYH missense mutation (R231L) that lies adjacent to the putative hMSH6 binding domain. The R231L mutant protein has severe defects in A/GO binding and in adenine glycosylase activities. The mutant fails to complement mutY-deficiency in Escherichia coli, but does not affect binding to hMSH6. These data support the role of the hMYH pathway in carcinogenesis. PMID- 17081687 TI - Adsorption of reactive dye onto carbon nanotubes: equilibrium, kinetics and thermodynamics. AB - The adsorption efficiency of carbon nanotubes for Procion Red MX-5B at various pHs and temperatures was examined. The amount adsorbed increased with the CNTs dosage; however, the adsorption capacity initially increased with the CNTs dosage (<0.25 g/l) and then declined as the CNTs dosage increased further (>0.25 g/l). The linear correlation coefficients and standard deviations of Langmuir and Freundlich isotherms were determined and the results revealed that Langmuir isotherm fitted the experimental results well. Kinetic analyses were conducted using pseudo first- and second-order models and the intraparticle diffusion model. The regression results showed that the adsorption kinetics were more accurately represented by a pseudo second-order model. Changes in the free energy of adsorption (DeltaG degrees ), enthalpy (DeltaH degrees ) and entropy (DeltaS degrees), as well as the activation energy (E(a)) were determined. DeltaH degrees and DeltaS degrees were 31.55 kJ/mol and 216.99J/molK, respectively, at pH 6.5 and 41.47 kJ/mol and 244.64 J/molK at pH 10. The activation energy was 33.35 kJ/mol at pH 6.5. DeltaH degrees, DeltaG degrees and E(a) all suggested that the adsorption of Procion Red MX-5B onto CNTs was by physisorption. PMID- 17081688 TI - Synthesis of ferrites obtained from heavy metal solutions using wet method. AB - Wet method was employed to the treatment of heavy metal-contaminated wastewater, and Zn(x)Fe(3-x)O(4), Ni(x)Fe(3-x)O(4) and Cr(x)Fe(3-x)O(4) (0Cr(3+) and the influence of the three ions on sample thermostability is Zn(2+)>Ni(2+)>Cr(3+). PMID- 17081689 TI - Involvement of mGluR5 in the ethanol-induced neuropathic pain-like state in the rat. AB - Alcohol neuropathy has been thought to involve decreased nerve function following chronic ethanol consumption. However, there is no reliably successful therapy, largely due to a lack of understanding of the central underlying mechanisms. The aim of this study was to investigate the mechanisms that contribute to the neuropathic pain-like state induced by chronic ethanol treatment in rats. Rats were chronically treated with ethanol diet (1.25-5% of ethanol) for over 70 days. Mechanical hyperalgesia was observed during ethanol consumption and even after ethanol withdrawal. Under these conditions, an immunohistochemical study showed an increase in metabotropic glutamate receptor 5 (mGluR5) immunoreactivity in the superficial spinal dorsal horn of chronic ethanol-fed rats. Furthermore, immunoblot analysis revealed that the protein level of mGluR5 was clearly increased following chronic ethanol consumption. These findings support the idea that the increased levels of mGluR5 in the spinal cord may be, at least in part, involved in the induction of ethanol-dependent neuropathic pain-like state. PMID- 17081690 TI - Visual mismatch negativity elicited by facial expressions under non-attentional condition. AB - A modified "cross-modal delayed response" paradigm was used to investigate putative processing of face expression in the absence of focused attention to the face. Neutral, happy and sad faces were presented during intervals occurring between a tone and a response imperative signal (a faint click), while subjects were instructed to discriminate the location of the tone as quickly and accurately as possible and to ignore the faces. A neutral face was presented in 80% of the trials whereas the happy and sad faces were presented in the remaining trials - 10%, respectively. Expression mismatch negativity (EMMN) was obtained by subtracting the ERP elicited by neutral faces from that elicited by sad faces or happy faces. The EMMN started from around 120 ms (sad) and 110 ms (happy) lasting up to 430 ms (sad) and 360 ms (happy) post-stimulus. The EMMN elicited by sad faces was more negative than that elicited by happy faces. Both EMMNs distributed over posterior areas and covered larger areas in the right than in the left hemisphere sites (especially for happy EMMN). PMID- 17081691 TI - Steroid requirements for regulation of the alpha4 subunit of the GABA(A) receptor in an in vitro model. AB - The alpha4 subunit of the GABA(A) receptor (GABAR) has relatively low expression in the CNS, but is increased in vivo following 48 h administration of the GABA modulatory steroid 3alpha-OH-5alpha[beta]-pregnan-20-one (THP or [allo]pregnanolone) to female rats. The purpose of the following study was to determine the optimal conditions for steroid-induced upregulation of alpha4 expression in an in vitro model. To this end, we used the IMR-32 cell, a neuroblastoma cell line, which normally expresses alpha4 mRNA at low levels. In undifferentiated IMR-32 cells, 48 h administration of THP increased alpha4 expression when ambient THP levels were reduced by the 5alpha-reductase blocker 4MA, suggesting that the background steroid milieu affects steroid regulation of this subunit. Following neuronal differentiation in serum-free medium, 48 h THP treatment significantly increased alpha4 expression two-fold following application of nerve growth factor (NGF) suggesting that development of neuronal processes facilitates this effect of the steroid. In the absence of NGF treatment, combined administration of 17beta-estradiol (E2) plus THP also increased alpha4 expression to a similar extent as THP following NGF treatment. In addition, E2 alone effectively increased alpha4 expression to maximal levels following NGF treatment. In contrast, neuronal differentiation in the absence of serum deprivation did not increase alpha4 levels. These results suggest that both THP and E2 can increase expression of the GABAR alpha4 subunit, but that this effect is dependent upon the background steroid milieu as well as the degree of neuronal development. These findings demonstrate optimal conditions for steroid induced upregulation of the alpha4 subunit in an in vitro system. PMID- 17081692 TI - Haematological and histopathological findings after ovariectomy in Trypanosoma cruzi infected mice. AB - The aim of this study was to investigate the influences of ovariectomy on histopathological and hematological parameters during the course of Trypanosoma cruzi infection. Hematological and immunological homeostasis is influenced by gonadal steroid hormones. Ovariectomy exerts profound influences on parasitic diseases including T. cruzi infection through modulation of the host's immune response. Three groups of female Mus musculus were infected with 4000 blood trypomastigotes of the Y strain of T. cruzi. One group was subjected to ovariectomy, another to simulated surgery before the infection, and a third group of unoperated animals were used as controls. Marked differences were detected in the responses of blood and tissue parasites. On day 9, post-infection parasitism was significantly higher in ovariectomized animals (P<0.05). These results were confirmed by histopathological studies, in which ovariectomized animals displayed hearts with higher number of amastigote burdens, increased inflammatory infiltrate, enhanced tissue fibers disorganization and decreased lytic antibody percentage, when compared to their counterparts. On day 9 the hematological changes were more apparent, with a decrease in erythrocytes, platelets and leucocytes for ovariectomized infected animals. Simulated surgery, as a stressful agent, did not cause any imbalance in parasitism or in the hemogram profile. The results confirm the importance of the female steroids in resistance against T. cruzi infection. PMID- 17081693 TI - New anti angiogenesis developments through electro-immunization: optimization by in vivo optical imaging of intradermal electro gene transfer. AB - Direct application of high voltage electric pulses of milliseconds duration to the skin of a mouse enhances in vivo intradermal delivery of injected therapeutic molecules such as DNA. The efficacy of gene transfer and expression is dependent on electrical parameters. DNA electrotransfer in tissues increases the associated DNA expression vaccine potency. This protocol is called "electro-immunization". In the present study, we report a new strategy for optimizing electro immunization. In vivo fluorescence imaging was used to detect the expression of a fluorescent protein (DsRed) and therefore allowed rapid optimization of the protocol. In vivo electrogenetransfer in the skin was well tolerated and DsRed expression was followed for over 2 weeks. Expression was voltage dependent under our conditions. Parameters were selected giving the highest level of expression. Under these optimized conditions, electrotransfer of a plasmid encoding VEGF was evaluated for its immune response as a gene therapy of interest involved in anti angiogenic strategies. Anti VEGF 165 antibodies in sera of mice were evaluated by ELISA and compared to those obtained after conventional immunization. Comparable titres of antibodies were obtained in both groups. An IgG2a predominance was found in mice immunized with the plasmid whereas a IgG1 predominance was observed in mice immunized classically. Skin electro-immunization is therefore shown as a good route for DNA immunization for anti-angiogenesis concern. PMID- 17081694 TI - Scalding as an unusual cause of pyoderma gangrenosum. PMID- 17081695 TI - Evidence profiles for breast cancer: benefit/harms data based on the totality of randomized evidence. PMID- 17081696 TI - Mice heterozygous for neurotrophin-3 display enhanced vulnerability to excitotoxicity in the striatum through increased expression of N-methyl-D aspartate receptors. AB - The striatum is one of the brain areas most vulnerable to excitotoxicity, a lesion that can be prevented by neurotrophins. In the present study, intrastriatal injection of the N-methyl-d-aspartate receptor (NMDAR) agonist quinolinate (QUIN) was performed in mice heterozygous for neurotrophin-3 (NT3 +/ ) or brain-derived neurotrophic factor (BDNF +/-) to analyze the role of endogenous neurotrophins on the regulation of striatal neurons susceptibility to excitotoxic injury. QUIN injection induced a decrease in dopamine- and cyclic AMP regulated phosphoprotein of 32 kDa (DARPP-32) protein levels that was higher in NT-3 +/- than in BDNF+/- or wild type animals. This enhanced susceptibility was specific for enkephalin- and tachykinin-positive projection neurons, and also for parvalbumin-positive interneurons. However the excitotoxic damage in large interneurons was not modified in NT-3 +/- mice compared with wild type animals. This effect can be related to the regulation of NMDARs by endogenous NT-3. Thus, our results show that there is an age-dependent regulation of NMDAR subunits NR1 and NR2A, but not NR2B, in NT-3 +/- mice. The deficit of endogenous NT-3 induced a decrease in NR1 and NR2A subunits at postnatal day (P) 0 and P3 mice respectively, whereas an upregulation was observed in 12 week old NT-3 +/- mice. This differential effect was also observed after administration of exogenous NT 3. In primary striatal cultures, NT-3 treatment induced an enhancement in NR2A, but not NR2B, protein levels. However, intrastriatal grafting of NT-3 secreting cells in adult wild type mice produced a down-regulation of NR2A subunit. In conclusion, NT-3 regulates the expression of NMDAR subunits modifying striatal neuronal properties that confers the differential vulnerability to excitotoxicity in projection neurons and interneurons in the striatum. PMID- 17081697 TI - Delayed synapsing muscles are more severely affected in an experimental model of MuSK-induced myasthenia gravis. AB - Myasthenia gravis can be induced in mice by injecting the extracellular domain of rat muscle-specific kinase (MuSK), a transmembrane receptor tyrosine kinase involved in agrin signaling at the neuromuscular junction. About 5-10% of human myasthenia gravis patients have autoantibodies against MuSK. Here we have examined mouse neuromuscular junctions following MuSK immunization in two groups of muscles that can be distinguished on the basis of the timing of neuromuscular synaptogenesis and their response to perturbation of agrin signaling. We used confocal microscopy to characterize the distribution and expression of nicotinic acetylcoline receptors and of two presynaptic makers, neurofilament protein and synaptophysin. We observed disruption of neuromuscular junctions in all muscles examined in this model of myasthenia gravis. However delayed-synapsing muscles, including the diaphragm, sternomastoid and tibialis posterior, were significantly more severely affected than fast-synapsing muscles, including the intercostal, adductor longus and tibialis anterior. These results suggest a basis for the differential susceptibility of muscles in different classes of myasthenia gravis patients, including patients with autoantibodies against MuSK. PMID- 17081698 TI - Rolipram: a specific phosphodiesterase 4 inhibitor with potential antipsychotic activity. AB - Currently available antipsychotic medications work primarily by antagonizing D2 dopamine receptors, thus raising intracellular cAMP levels. We hypothesized that intracellular stimulation of cAMP levels in the CNS would have similar effects to treatment with antipsychotic medication. To test this hypothesis, we studied the effect of an acute treatment of rolipram, an inhibitor of type 4 phosphodiesterases that degrade cAMP, on acoustic startle and prepulse inhibition (PPI) of the acoustic startle response in C57BL/6J mice known to exhibit poor PPI. PPI is disrupted in schizophrenia patients, and the ability of a drug to increase PPI in mice is predictive of antipsychotic efficacy. We show here that acute treatment with rolipram significantly increases PPI at doses that do not alter the acoustic startle response (lowest effective dose 0.66 mg/kg). In addition, rolipram (0.66 mg/kg) blocks the disruptive effects of amphetamine (10 mg/kg) on PPI. At a slightly higher dose (1.0 mg/kg), rolipram also induces catalepsy. Thus, phosphodiesterase-4 (PDE4) inhibition has many of the same behavioral effects as traditional antipsychotic medications. In contrast to traditional antipsychotics, these effects are achieved through alteration of an intracellular second messenger system rather than antagonism of neurotransmitter receptors. Given previous reports showing rolipram improves cognition, we conclude that PDE4 represents an important novel target for further antipsychotic drug development. PMID- 17081699 TI - HPA- and HPT-axis alterations in chronic posttraumatic stress disorder. AB - Posttraumatic stress disorder (PTSD) has been associated with dysregulation of the hypothalamus-pituitary-adrenal (HPA) axis as well as of the hypothalamus pituitary-thyroid (HPT) axis. Findings have not been consistent and may depend on methodological issues like controlling for relevant variables. This study examines the levels of six HPA and HPT-axis related hormones in civilian PTSD patients without psychotropic medication. In a cross sectional study, 39 chronic PTSD patients and 44 healthy volunteers were included. Psychometric instruments included SCID, SI-PTSD, IES-R and BDI. The plasma hormones levels assessed were cortisol, dehydroepiandrosterone (DHEA), and dehydroepiandrosterone sulfate (DHEA S), prolactin, thyrotropin (TSH), and free thyroxin (fT4). Results showed that patients had significantly lower plasma cortisol, prolactin and TSH levels compared to the comparison group. The difference between TSH levels in patients and comparison subjects only emerged after controlling for relevant background variables. Furthermore, the severity of PTSD symptoms was negatively related to cortisol levels. Secondary analyses revealed no statistically significant effect of comorbid depression (26% of patients) on any of the hormone levels. Complex feedback mechanisms are likely to result in altered levels of stress related hormones in PTSD, and results depend on controlling for relevant variables. Further research with longitudinal designs is needed to find out whether these lower hormone levels are preexisting risk factors or consequence of trauma and whether these alterations are deleterious or adaptive. PMID- 17081701 TI - Cortisol responses to psychological stress in adults after prenatal exposure to the Dutch famine. AB - Experimental studies in animals show that prenatal undernutrition leads to lifelong alterations in the activity of the hypothalamic-pituitary-adrenal (HPA) axis. Some studies have shown associations between low birth weight and an increased HPA response to psychological stress. We tested the hypothesis that prenatal exposure to the Dutch 1944-1945 famine leads to an elevated HPA response to psychological stress in adult life. We measured salivary cortisol responses to a psychological stress protocol among 694 adults who were born as term singletons in Amsterdam, the Netherlands, around the time of the 1944-1945 Dutch famine. We compared cortisol profiles of participants exposed to famine during late (n=120), mid (n=100), or early gestation (n=62) to profiles of participants unexposed to famine during gestation (n=412). The mean increase in cortisol concentrations from baseline was 30% (95% CI 23-37). There were no statistically significant differences in the mean profile of cortisol response to the psychological stress protocol between participants exposed and unexposed to famine in utero. The mean sex and BMI adjusted difference in cortisol response for those exposed compared to those unexposed was -6% (95% CI: -15 to 2). The cortisol profiles of those exposed in late (-4% [95% CI: -16 to 7]), mid (-9% [95% CI: -22 to 3]) or early gestation (-4% [95% CI: -20 to 10]) did not differ from the profile of those unexposed to famine. We conclude that prenatal exposure to famine does not seem to be associated with the response of the HPA axis to psychological stress. However, the stress protocol we have used may have been unsuccessful in inducing a strong enough HPA axis activation to be able to detect famine related differences. PMID- 17081700 TI - Stress sensitivity in metastatic breast cancer: analysis of hypothalamic pituitary-adrenal axis function. AB - The normal diurnal cortisol cycle has a peak in the morning, decreasing rapidly over the day, with low levels during the night, then rising rapidly again to the morning peak. A pattern of flatter daytime slopes has been associated with more rapid cancer progression in both animals and humans. We studied the relationship between the daytime slopes and other daytime cortisol responses to both pharmacological and psychosocial challenges of hypothalamic-pituitary-adrenal (HPA) axis function as well as DHEA in a sample of 99 women with metastatic breast cancer, in hopes of elucidating the dysregulatory process. We found that the different components of HPA regulation: the daytime cortisol slope, the rise in cortisol from waking to 30 min later, and cortisol response to various challenges, including dexamethasone (DEX) suppression, corticotrophin releasing factor (CRF) activation, and the Trier Social Stress Task, were at best modestly associated. Escape from suppression stimulated by 1mg of DEX administered the night before was moderately but significantly associated with flatter daytime cortisol slopes (r=0.28 to .30 at different times of the post DEX administration day, all p<.01). Daytime cortisol slopes were also moderately but significant associated with the rise in cortisol from waking to 30 min after awakening (r=.29, p=.004, N=96), but not with waking cortisol level (r=-0.13, p=.19). However, we could not detect any association between daytime cortisol slope and activation of cortisol secretion by either CRF infusion or the Trier Social Stress Task. The CRF activation test (following 1.5mg of DEX to assure that the effect was due to exogenous CRF) produced ACTH levels that were correlated (r=0.66, p<.0001, N=74) with serum cortisol levels, indicating adrenal responsiveness to ACTH stimulation. Daytime cortisol slopes were significantly correlated with the slope of DHEA (r=.21, p=.04, N=95). Our general findings suggest that flatter daytime cortisol slopes among metastatic breast cancer patients may be related to disrupted feedback inhibition rather than hypersensitivity in response to stimulation. PMID- 17081702 TI - Visual evoked potentials in chronic alcoholism. AB - In order to evaluate the effect of alcohol consumption and the effect of abstinence on central nervous system generated parameters, we performed pattern visual evoked potential (PVEP) recordings on chronic alcoholics. The study was conducted on forty patients diagnosed as chronic alcoholics according to DSM IV criteria. They were aged mean: 42, and had histories of alcohol abuse for at least six years (mean: 21). 15% of the patients demonstrated abnormal VEP results at least in one tested eye. In order to test the effect of abstinence period on P100 latency values, alcoholics were divided in to two subgroups. Group I (Gr I) consisted twenty-four alcoholics who had been abstinent for less than thirty days (mean: 14), and Group II (Gr II) consisted sixteen alcoholics who had been abstinent for more than thirty and less than seventy-six days (mean: 38) The mean P100 latency of Gr I and Gr II was 101 and 102 milliseconds (ms) respectively; and when compared to normal controls the difference was statistically significant (p: 0.016, p: 0.009). Abnormal VEP in asymptomatic chronic alcoholics suggests that they may be useful in the detection of early changes and in following the progress of patients with the disorder. PMID- 17081703 TI - Determining a cut-off on the Severity of Dependence Scale (SDS) for alcohol dependence. AB - Optimal cut-off points on the Severity of Dependence Scale (SDS) indicative of clinically significant dependence have been determined for a range of substance types. This study aims to determine a cut-off point on SDS that discriminates between the presence and absence of a DSM-IV diagnosis of alcohol dependence. A structured interview was administered to 90 alcohol users in Sydney, Australia. Receiver Operating Characteristic curve analysis confirmed the utility of the SDS alcohol for characterising and diagnosing persons with respect to their alcohol dependent status to an accuracy of 85%. A SDS score of 3 or above was determined as optimal for characterising alcohol dependence. Evidence is also provided confirming that the SDS-alcohol is a valid, reliable uni-dimensional scale for measuring alcohol dependence. It has been demonstrated that the SDS-alcohol can be used to characterise an individual's alcohol-dependent status. A cut-off value for SDS-alcohol provides additional meaning and value to the scale for clients and clinicians and will enable researchers to characterise the prevalence of alcohol dependence in their target populations. PMID- 17081704 TI - p21 as a prognostic factor in non-small cell lung carcinomas. AB - Few studies have focused on the correlation between p21 expression and survival for patients with non-small cell lung carcinoma (NSCLC), and the results are not consistent. We investigated the expression of p21 in 90 cases of NSCLC to evaluate the correlation between the p21 expression level and the clinicopathologic characteristics with patient survival. p21 was expressed in the nuclei of all the NSCLCs. The percentage of immunoreactive cells varied from 1% to 70%. All the patients were subdivided into a high and a low p21 expression group on the basis of the median percentage (17.05). There was no significant correlation between the p21 expression level and age, gender, histologic type, histologic grade, or stage. Using uni- and multivariate analyses, survival was significantly associated with gender, stage, and the p21 expression level. The survival rate for the high p21 expression group was higher than that for the low p21 expression group in the entire patient group, and especially for stage II and III patients, males, adenocarcinomas, or p53-positive tumors. Our findings showed that high p21 expression was an independent prognostic factor for NSCLC. p21 may be useful for determining the candidates for adjuvant therapies and also for selecting the appropriate chemotherapeutic agents. PMID- 17081705 TI - The consequences of surrendering a degree of freedom to the participant in a contingency assessment task. AB - Many studies of contingency judgments have used a task in which, on each trial, the participant is free either to respond or not to respond, and an outcome may, or may not, be presented. Typically, the experimenter specifies a nominal value for the contingency between responding and outcome, but the actual values of a variety of variables experienced by a particular participant depend on that participant's frequency of responding. The results of computer simulations of various strategies for implementing the contingency manipulation, and the results of an experiment, indicate that the same nominal contingency value will lead to considerable variability in the actual contingency experienced by participants. Moreover, nominal contingency manipulations are confounded with the probability that the subject experiences an outcome. While researchers might be aware of these issues, not enough attention has been paid to their potential impact. PMID- 17081706 TI - Testosterone directly induces progesterone production and interacts with physiological concentrations of LH to increase granulosa cell progesterone production in laying hens (Gallus domesticus). AB - Blocking testosterone action with immunization or with a specific antagonist blocks the preovulatory surge of progesterone and ovulation in laying hens. Thus, testosterone may stimulate progesterone production in a paracrine fashion within the ovary. To test this hypothesis, we evaluated the effects of testosterone and its interaction with LH on the production of progesterone by granulosa cells in culture. Hen granulosa cells obtained from preovulatory follicles were cultured in 96 well plates. The effects of testosterone (0-100ng/ml) and/or LH (0 100ng/ml) were evaluated. LH-stimulated progesterone production in a dose response manner up to 10ng/ml (p<0.01). Testosterone, up to 10ng/ml, increased progesterone production in a dose response manner in the absence of LH and at all doses of LH up to 1ng/ml (p<0.001). However, at supraphysiological concentrations of LH (10 and 100ng/ml) there was no further increase in progesterone production caused by testosterone (p>0.05). Finally, the addition of 2-hydroxyflutamide (0 1000mug/ml) to hen granulosa cells cultured with 10ng/ml of testosterone reduced progesterone production in a dose response manner (p<0.001). In conclusion, testosterone stimulates progesterone production in preovulatory follicle granulosa cells and interacts with physiological concentrations of LH to increase progesterone production. In addition, testosterone stimulation on granulosa cells is specific since the testosterone antagonist decreased testosterone stimulatory action. PMID- 17081708 TI - In vitro and in vivo evaluation of carbamazepine-loaded enteric microparticles. AB - The objective of the study was to prepare and evaluate carbamazepine-loaded enteric microparticles produced by a novel coacervation method. An aqueous polymeric stabilizer solution was added to an organic carbamazepine/Eudragit L100 55 solution. Water, which is a non-solvent for the drug and the enteric polymer, caused phase separation and the formation of coacervate droplets. These droplets hardened into microparticles upon further addition of the aqueous phase. The microparticles were characterized with respect to particle size distribution, morphology, encapsulation efficiency, yield, physical state and physical stability of the drug, wettability, in vitro release and in vivo bioavailability. Microparticles with a smooth surface and dense structure were obtained with high encapsulation efficiency (>85%) and yield (>90%). The drug was in a non crystalline state in the matrix and physically stable for 5 months at room temperature. Under sink conditions, the drug dissolution rate from the microparticles was significantly enhanced compared to the physical mixture and to the pure drug; the release profile of the microparticles was stable after 5 months. Under non-sink conditions, an unstable supersaturated solution of carbamazepine was obtained from microparticles with the subsequent formation of needle-shaped crystals. The high surface area and good wettability of the microparticles, the non-crystalline state of the drug in the matrix and the fast dissolution rate contributed to a significantly enhanced oral bioavailability from the microparticles when compared to the physical mixture. PMID- 17081707 TI - Effects of in vitro maturation of monkey oocytes on their developmental capacity. AB - The study of in vitro maturation (IVM) of rhesus monkey oocytes has important implications for biomedical research and human infertility treatment. In vitro matured rhesus monkey oocytes show much less developmental potential than IVM oocytes of other species. Since about 1980 when rhesus monkey IVM, in vitro fertilization (IVF) and in vitro embryo culture (IVC) systems were established, numerous efforts have been made to improve the developmental competence of oocytes and to understand the mechanisms regulating oocyte maturation. This review describes recent progress in this area, particularly the effects of factors such as steroid hormones, energy substrates, amino acids, ovarian follicle status, maternal age and breeding season on the developmental competence, gene expression patterns and genome integrity of rhesus IVM oocytes. PMID- 17081709 TI - Sustained-release tablets of indomethacin-loaded microcapsules: preparation, in vitro and in vivo characterization. AB - Indomethacin (IDM) was encapsulated in gelatin-cellulose acetate phthalate (CAP) microcapsules (A) by complex coacervation method and in CAP microcapsules (B) by simple coacervation method. Microcapsules A and B, having mean diameters of 38.24 and 35.74 microm, respectively, were used to prepare sustained-release tablets A and B. The activation energy of thermal degradation for tablets A and B was calculated based on differential scanning calorimetry (DSC) to be 258.9 and 284.8 kcal/mol, respectively. In vitro release profiles showed no burst effect and release t(1/2) of the two sustained-release tablets were found to be 41.30+/-1.86 and 33.25+/-2.84 min, respectively, while that of IDM plain tablets C was 6.30+/ 0.39 min (P<0.01). In vitro release of IDM from tablets A and B could be described by Higuchi equation and zero-order kinetics, respectively. After per os (po) administration with physiological saline, their irritation to rat stomach was obviously reduced in comparison with tablets C. Pharmacokinetic study in rabbits showed that t(max) was delayed and C(max) lowered compared with tablets C and the values of AUC(0-24 h) of the three tablets were very close. PMID- 17081710 TI - Distribution, transition, adhesion and release of insulin loaded nanoparticles in the gut of rats. AB - The purpose of this work was to investigate distribution, transition, bioadhesion and release behaviors of insulin loaded pH-sensitive nanoparticles in the gut of rats, as well as the effects of viscosity agent on them. Insulin was labeled with fluorescein isothiocyanate (FITC). The FITC-insulin solution and FITC-insulin nanoparticle aqueous dispersions with or without hydropropylmethylcellulose (HPMC, 0.2%, 0.4%, or 0.8% (w/v)) were orally administered to rats, respectively. The amounts of FITC-insulin in both the lumen content and the intestinal mucosa were quantified by a spectrofluorimeter. The release profiles in the gut were plotted by the percentages of FITC-insulin released versus time. FITC-insulin nanoparticle aqueous dispersion showed similar stomach but lower intestine empty rates, and enhanced intestinal mucosa adhesion in comparison with FITC-insulin solution. Addition of the HPMC reduced the stomach and intestine empty rates, enhanced the adhesion of FITC-insulin to the intestine mucosa. The release of FITC-insulin from nanoparticles in the gut showed an S-shape profile, and addition of HPMC prolonged the release half-life from 0.77 to 1.51h. It was concluded that the behaviors of pH-sensitive nanoparticles tested in gastrointestinal tract of rats and the addition of HPMC were favorable to the absorption of the drug loaded. PMID- 17081711 TI - Pharmacodynamics of adenovector distribution within the inner ear tissues of the mouse. AB - Recent studies have demonstrated that delivery of genes to the inner ear can achieve a variety of effects ranging from support of auditory neuron survival to protection and restoration of hair cells, demonstrating the utility of vector based gene delivery. Translation of these findings to useful experimental systems or even clinical applications requires a detailed understanding of the pharmacokinetics of gene delivery in the inner ear. Ideal gene delivery systems will employ a well tolerated vector which efficiently transduces the appropriate target cells within a tissue, but spare non-target structures. Adenovectors based on serotype 5 (Ad 5) are commonly used vectors, are easy to construct and have a long track record of efficacious gene transfer in the inner ear. In this study we demonstrate that distribution of Ad5 vector occurs in a basal to apical gradient with rapid distribution of vector to the vestibule after delivery via a round window cochleostomy. Transduction of the vector and expression of the delivered transgene occurs by 10 min post vector delivery. At 24 h post delivery only 16% of vector that was initially detectable within the inner ear by quantitative PCR remained. Perilymph sampling was used to determine that vector concentrations in perilymph peaked at 30 min post delivery and then declined rapidly. Understanding these basic distribution patterns and parameters for delivery are important for the design of gene delivery vectors and vital for modeling dose responses to achieve safe efficacious delivery of a therapeutic agent. PMID- 17081712 TI - Complex tone processing and critical band in the human auditory cortex. AB - Psychophysical experiments in humans have indicated that the auditory system has a well-defined bandwidth for resolution of complex stimuli. This bandwidth is known as the critical bandwidth (CBW). Physiological correlates of the CBW were examined in the human auditory cortex. Two- and three-tone complexes were used as the sound stimuli with all signals presented at 55 dB sound pressure level (SPL). The duration of stimulation was 500 ms, with rise and fall ramps of 10 ms. Ten normal-hearing subjects took part in the study. Auditory-evoked fields were recorded using a 122-channel whole-head magnetometer in a magnetically shielded room. The latencies, source strengths, and coordinates of the N1m waves, which were found above the left and right temporal lobes approximately 100 ms after the onset of stimulation, were analyzed. The results indicated that N1m amplitudes were approximately constant when the frequency separation of a two-tone complex or the total bandwidth of a three-tone complex was less than the CBW; however, the N1m amplitudes increased with increasing frequency separation or total bandwidth when these were greater than the CBW. These findings indicate critical band-like behavior in the human auditory cortex. The N1m amplitudes in the right hemisphere were significantly greater than those in the left hemisphere, which may reflect a right-hemispheric dominance in the processing of tonal stimuli. PMID- 17081714 TI - Immune cell recruitment following acoustic trauma. AB - Acoustic trauma induces cochlear inflammation. We hypothesized that chemokines are involved in the recruitment of leukocytes as part of a wound healing response. The cochleas of NIH-Swiss mice, exposed to octave-band noise (8-16 kHz, at 118 dB) for 2h, were examined after the termination of exposure. Leukocytes were identified immunohistochemically with antibodies to CD45 and F4/80. Gene array analysis followed by RT-PCR was performed on cochlear tissue to identify up regulation of chemokine and adhesion molecule mRNA. The expression of the adhesion molecule ICAM-1 was also investigated immunohistochemically. Few CD45- or F4/80-positive leukocytes were observed in the non-exposed cochlea. Following acoustic trauma however, the number of CD45-positive cells was dramatically increased especially after 2 and 4 days, after which time the numbers decreased. F4/80-positive cells also increased in number over the course of a week. Gene array analysis indicated increased expression of monocyte chemoattractant protein 5 (MCP-5), monocyte chemoattractant protein 1 (MCP-1), macrophage inflammatory protein-1beta (MIP-1beta) and ICAM-1. RT-PCR, performed using primers for the individual mRNA sequences, confirmed the increased expression of MCP-1, MCP-5, MIP-1beta, and ICAM-1 relative to non-exposed mice. In the normal cochlea, ICAM-1 immunohistochemical expression was observed in venules, spiral ligament fibrocytes and in endosteal cells of the scala tympani. Expression increased to include more of the spiral ligament and endosteal cells after acoustic trauma. A cochlear inflammatory response is initiated in response to acoustic trauma and involves the recruitment of circulating leukocytes to the inner ear. PMID- 17081713 TI - Characterization of supporting cell phenotype in the avian inner ear: implications for sensory regeneration. AB - The avian inner ear possesses a remarkable capacity for the regeneration of sensory receptors after acoustic trauma or ototoxicity. Most replacement hair cells are created by renewed cell division within the sensory epithelium, although some new hair cells may also arise through nonmitotic mechanisms. Current data indicate that epithelial supporting cells play an essential role in regeneration, by serving as progenitor cells. In order to become progenitors, however, supporting cells may need to undergo partial dedifferentiation. In this review, I describe molecules that are expressed by supporting cells in the avian ear. Although a number of these molecules are likely to be critical to the maintenance of the supporting cell phenotype, we presently know very little about phenotypic changes in supporting cells during the early phase of regeneration. PMID- 17081715 TI - Investigation of biological activity of Lamii albi flos extracts. AB - Methanol, ethyl acetate and heptane extracts of Lamium album L. (Lamiaceae) were tested for the toxicity or ability to stimulate growth of human skin fibroblasts (HSF) in vitro. The biological effect depended on the HSF cells density and on the kind and concentration of extract that was used. At a density of 1x10(5) HSF cells/mL no cytotoxicity of extracts was observed during 24 h of incubation. During 24-72 h of incubation with a low (2x10(4) HSF cells/mL) density of cells, significant cytotoxicity was observed for methanol and ethyl acetate extracts at concentrations greater than 125 microg/mL. At concentration of 25 microg/mL the cells remained intact. However, when the cells were incubated with the heptane extract, there was observed relative high viability (>60%) of cells and significant, gradually increasing in time, cellular mitochondrial dehydrogenase activity, measured by MTT assay. Stimulation of human skin fibroblasts proliferation by the heptane extract of Lamii albi flos may indicate for its usefulness of wound healing in vivo and probability of perspectives its use in designing new medicinal plant preparations. In order to determine the biologically active compounds, the plant extracts were separated by high performance thin layer chromatography (HPTLC) on silica gel Si 60 F(254) combined with densitometry. Phenolic acids were investigated by high performance liquid chromatography (HPLC). PMID- 17081716 TI - Differential accumulation of pulmonary and cardiac mast cell-subsets and eosinophils between fatal anaphylaxis and asthma death: a postmortem comparative study. AB - The distribution profile of infiltrated mast cell-subpopulations and eosinophils in the lung and heart sections of the patients who died of severe allergic hyperresponsiveness, was investigated. Four study groups were designed comprising 9 cases who died in systemic anaphylaxis (Group I), 10 asthmatic individuals whose death were assigned to acute and severe bronchial asthma (Group II), 10 asthmatic cases who died from non-immunological diseases (Group III). Twenty consecutive autopsies of non-allergic subjects who died of unnatural causes (Group IV) served as control group in this study. Utilizing antibodies against human tryptase and chymase and a double immunohistochemical staining method, we distinguished successfully all three subsets of mast cells (MC), MC-TC (containing both tryptase and chymase), MC-T (containing only tryptase) and MC-C (containing only chymase) types, subdivided on the basis of the protease compositions of their secretory granules. In order to immunostaining eosinophils, we used antibody to major basic protein as a marker. We also measured postmortem blood tryptase, specific and total serum IgE. The intriguing finding of this study was the marked differences of cellular composition in the lung between fatal anaphylaxis and asthma death. Significant augmentation of MCs infiltrated in lung and heart sections of anaphylaxis patients and drastic infiltration of bronchial eosinophils in asthmatic death and consequent release of their related inflammatory mediators might explain the differential expression of the associated symptoms in these two groups. The anaphylactic deaths did show neither emphysema nor significant mucous bronchial secretions whereas all asthmatic deaths did. The degree of pulmonary congestion and edema was also more severe in anaphylaxis. This corresponded with the histological findings and the location and number of mast cell-subsets and eosinophils in the different compartments of the lungs. We have demonstrated that the third type of mast cell MC-C is only found in the lungs in anaphylactic deaths. The practical consequence of our study will be that it is now possible to confirm a suspicion of anaphylaxis death not only by measurements of serum mast cell tryptase, but also by immunohistochemical methods. PMID- 17081717 TI - Rapid on-line extraction and quantification of escitalopram from urine using sol gel columns and mass spectrometric detection. AB - A fast and easy way to quantify escitalopram in urine has been developed. A capillary with a silica based monolithic bed inside is used to extract escitalopram from urine and the for mass spectrometry detrimental matrix is washed away by applied pressure. The analyte is eluted by a solution containing organic modifier and directly electrosprayed into a time-of-flight mass spectrometer, ESI-TOF-MS. This method makes it possible to load large volumes of sample onto the column and preconcentrate escitalopram on-line before detection. Standard addition of escitalopram to the urine sample gave a linear calibration curve (R(2)=0.988). The analyzed sample was found to contain an escitalopram concentration of 0.62 ng/ml, well in line with earlier publications. The calculated LOD was 10 pg/ml and LOQ was 34 pg/ml as compared to earlier reports with a LLOQ of 1 ng/ml. The intra day variation of the escitalopram peak area is less than 6.3%. PMID- 17081718 TI - Methodology of the thyroid gland disease decision-making using profiling in steroid hormone pathway. AB - To find out the genetic factors of outbreak of thyroid gland disease, we developed the thyroid gland decision-making system, which processes the metabolic profile in steroid hormone map using a statistical method. Metabolic profile is a measured data of lots of mixed materials that includes not only known metabolites, but also unknown ones, which is estimated to have an influence on the thyroid gland disease. Therefore, to develop thyroid gland disease decision making system, analyzing metabolic profile containing multi-materials would be useful for diagnosing thyroid gland disease. Because experimental values used for system construction are area values for the retention time, the observations are preprocessed through variable transition and t-test to use the area values concurrently and the highly correlated materials are estimated by principal component analysis. The thyroid gland decision-making system developed through the logistic regression is an excellent system demonstrating 98.7% accuracy in the classification table. PMID- 17081719 TI - Public understanding of growth charts: a review of the literature. AB - OBJECTIVE: To examine current knowledge about the general public's comprehension of growth charts. METHODS: A literature search was carried out to identify and analyze the findings of studies that have examined the extent to which non healthcare professionals comprehend the information presented by growth charts. RESULTS: Few studies have examined how well parents understand growth charts. These studies have mostly been conducted in developing countries and have yielded inconclusive results. CONCLUSION: Growth charts can be useful for such purposes as growth monitoring and public health education, but their effectiveness as an educational tool depends on whether parents and caregivers comprehend the information they offer. The literature has not clearly proven that growth charts are easily understood by the general population. PRACTICE IMPLICATIONS: Further studies of the public's understanding of growth charts are needed to help guide health care providers in their use of growth charts as educational tools. PMID- 17081720 TI - Effective chronic disease management: patients' perspectives on medication related problems. AB - OBJECTIVE: To examine medication-related problems from the perspective of patients with a chronic condition and to identify how they may be supported in managing their medication. METHODS: Patients prescribed medication for cardiovascular disease were recruited through five general medical surgeries and four community pharmacies in south London. Data were collected in 98 face-to-face interviews in participants' own homes. Interviews were designed to enable a detailed and holistic exploration of medication-related problems from participants' perspectives. Data were audio-recorded and transcribed verbatim to allow qualitative analysis. RESULTS: Five broad categories of medication-related problem emerged which were examined in the context of patients' perspectives on, and experiences of, the use of medicines and health services. These were concerns about and management of side effects; differing views regarding the use of medicines; cognitive, practical and sensory problems; lack of information or understanding; and problems with access to, and organisation of, services. CONCLUSION: All categories of problem had potential implications for the success of therapy in that they created barriers to adherence, access to medication or informed decision-making. The study demonstrated how patients actively engage in decision-making about their medicines in the home, if not in the consultation. PRACTICE IMPLICATIONS: The five categories of problem provide a focus for interventions by health professionals to support patients in achieving optimal theory outcomes. They demonstrate the need for a comprehensive approach, spanning patient education to the systems of delivery of care. Within the NHS in Britain, policy and practice initiatives are being designed to achieve this end. Further research should focus on the evaluation of professional practices and service developments in supporting patients in the self-management of their medicines. PMID- 17081721 TI - Pancreaticoduodenectomy with a "twist": the challenges of pancreatic resection in the presence of situs inversus totalis and situs ambiguus. PMID- 17081722 TI - Patterns of lateral pelvic lymph node metastases and micrometastases for patients with lower rectal cancer. AB - AIMS: We aimed at investigating the patterns of lymph node metastases and micrometastases in regions of lateral pelvic area, examining circumferential margin involvement and clarifying their prognostic significance. METHODS: Large tissue slice and tissue array were adopted in the study of 67 patients with AJCC stages I-III lower rectal cancer who underwent total mesorectal excision with systematic lateral pelvic dissection. The outcomes were followed. RESULTS: Altogether, 726 lateral lymph nodes were examined, with 32 and 38 were involved by tumor metastases and micrometastases, respectively. Fifty-eight (82.9%) of the involved lymph nodes were smaller than 5mm. Status of lateral nodes was related to that of mesorectal ones. Middle rectal root (45.5%), internal iliac (31.8%) and obturator (22.7%) regions were more likely to be involved by metastases. Patients with lateral metastases, similar to the group with micrometastases, suffered more recurrence and poorer survival when compared with the ones without metastases. The occurrence of circumferential margin involvement suggested poor prognosis and was related to lateral node status. CONCLUSIONS: In lateral pelvic area, the majority of lymph nodes harboring tumor were small and could easily be neglected by conventional examination. Incidence of lateral metastases differed among regions, thus more attention should be given to the clearance of the highly occurred areas. More extensive range of dissection and/or adjuvant therapy was recommended for patients with lateral node metastases, micrometastases and circumferential margin involvement, since they predisposed poor prognosis. PMID- 17081723 TI - Methylene blue dye versus combined dye-radioactive tracer technique for sentinel lymph node localisation in early breast cancer. AB - AIM: The study compared the accuracy and success rate of two techniques, methylene blue alone versus combined methylene blue and radioactive colloid in sentinel lymph node localisation in the management early breast cancer. METHODS: Three hundred and twenty-nine patients with tumours less than 2 cm on ultrasound assessment were prospectively evaluated. One hundred and seventy-three patients (Group A) underwent sentinel lymph node localisation using 1 ml of 1% methylene blue. A combined technique of both methylene blue and radioactive colloid was used in 156 patients (Group B). Application of both was subdermal and subareolar. Sentinel lymph nodes were examined by standard microscopy. Patients underwent breast conservation surgery or mastectomy and sentinel node guided four node axillary sampling+/-clearance. RESULTS: In Group A, the sentinel lymph node identification rate was 96.5%. The negative predictive value was 96.3%, with false negative of 3.7% and accuracy of 87.4%. In group B the identification rate for sentinel lymph node was 98.7%, with false negative of 4.1%, negative predictive value of 96%, and accuracy of 83.8%. CONCLUSION: Sentinel lymph node localisation using methylene blue or combined dye and radioactive tracer technique predicts the axillary lymph node status in early breast cancer with comparable success rates, accuracy and false negative rates. The combined technique facilitates quicker identification of sentinel lymph node; however the dye technique alone can be used successfully in centres without nuclear medicine facilities. PMID- 17081724 TI - Histopathological outcome of 597 isolated soft tissue tumors suspected of soft tissue sarcoma: a single-center 12-year experience. AB - BACKGROUND: The aim of this present report was to analyze the patients referred to us with the presumptive diagnosis of soft tissue sarcoma (STS). METHODS: We reviewed all patients referred to us with suspected soft tissue sarcoma (STS) of the extremities or trunk over a 12-year period. RESULTS: We treated 597 patients with soft tissue tumors. Open biopsy revealed soft tissue sarcoma in 318 cases, benign mesenchymal tumor in 124 cases and isolated metastases (ISTM) from carcinomas in 98 patients; other pathologies were found in 57 patients. The primary carcinomas were lung cancer in 26 patients, breast cancer in 19 patients, renal carcinoma in 16 patients, carcinoma of the esophagus in 12 patients, colonic carcinoma in 5 patients, thyroid gland cancer in 6 patients, and in 14 patients carcinoma of unknown primary was diagnosed. CONCLUSIONS: In our collective with soft tissue tumor, 50% of the patients had the diagnosis of soft tissue sarcoma, 20% presented with a metastasis of carcinoma and 20% had a benign tumor. Referring to our results, in patients with the presumptive diagnosis of soft tissue sarcomas, soft tissue metastasis of a primary carcinoma was unexpectedly common, indicating that greater consideration should be given to this differential diagnosis. PMID- 17081725 TI - Long-term prognosis of primary retroperitoneal soft tissue sarcoma. AB - AIMS: To evaluate the result of treatment and long-term outcome of a population based cohort of patients with retroperitoneal soft tissue sarcoma (RSTS). METHODS: Between 1 January 1989 and 1 January 1994, 143 patients diagnosed as having primary RSTS were selected from a national pathology database (PALGA) in the Netherlands. In this population-based group of patients, the result of surgery, overall survival (OS) and disease-free survival (DFS) were analysed as well as factors affecting OS and DFS. Median follow-up was 10.2 years. RESULTS: Operative treatment resulted in a complete tumour resection in 55% of the patients (n=78), low- and intermediate-grade tumours were more often completely resected than high-grade tumours (P=0.016). Five- and 10-year cumulative OS was 39% and 21%, respectively, while DFS was 22% and 17%, respectively. In a multivariate analysis low malignancy grade (P=0.017) and a complete tumour resection (P<0.001) were associated with better OS. CONCLUSIONS: Complete tumour resection and low malignancy grade were independent favourable prognosticators. However, these factors were related too, since surgical success was influenced by malignancy grade. PMID- 17081726 TI - [Organization of urgent blood transfusion in France: analysis of practice patterns in centers involved in care delivery to multiple trauma patients]. AB - OBJECTIVE: To describe the organization of medical and transfusion services within the framework of hospital admission for multiple trauma. STUDY DESIGN: National survey in France. MATERIAL AND METHODS: All French hospitals caring for multiple trauma patients were sent a questionnaire. Organization of the medical and transfusion services, and their related resources were evaluated. RESULTS: Among 372 questionnaires sent, 116 replies were received from structures which care for 18 (1-500) multiple trauma patients each year. An orthopaedic and an abdominal surgery unit were widely available whereas a neurosurgeon was available in 21% of responding centers. A transfusion site was found in 43%, whereas others have either a deposit for distribution to specific patients (40%) or a small deposit to cover urgent situations (17%). Comparison with legal or expert based rules of adequate transfusion process disclosed a variable incidence of practice dysfunctions (2-49%) depending on the parameter assessed. CONCLUSION: The French organization of multiple trauma patients' care and blood transfusion delivered for these patients is not homogenous. Dysfunctions were found in all types of hospitals. Recommendations describing good practice seem necessary to be built at the national level. PMID- 17081727 TI - Role of the universal stress protein UspA of Salmonella in growth arrest, stress and virulence. AB - Pathogenic bacteria employ a variety of mechanisms to resist a barrage of stresses they encounter during active growth in or outside the host as well as during growth stasis. An in silico screen of the Salmonella genome sequence revealed that Salmonella typhimurium LT2 possesses a homologue belonging to the universal stress protein A (UspA) family. We assessed the transcriptional profile of uspA in S. typhimurium C5 by constructing a lacZ fusion revealing that uspA is induced by metabolic, oxidative, and temperature stresses. The highest transcriptional levels occurred in cells entering stationary phase, an observation consistent with expression patterns in Escherichia coli. The protein was purified as a fusion with GST (UspA(F)) and antibodies raised against UspA(F) revealed elevated protein levels in stressed and growth-arrested cells. Inactivation of uspA in S. typhimurium C5, lead to increased susceptibility to stress conditions. Furthermore, UspA makes an important contribution to the in vivo virulence of Salmonella in mice thus highlighting the importance of stress resistance regulation in pathogenicity and survival within the host. PMID- 17081728 TI - Attention as a controller. AB - We investigate, by constructing suitable models, the manner in which attention and executive function are observed to interact, including some aspects of the influence of value/emotion on this interaction. Attention is modelled using the recent engineering control model (Corollary Discharge of Attention Movement, CODAM), which includes suitable working memory components. We extend this model to take account of various executive functions performed in working memory under attention control, such as rehearsal, substitution and transformation of buffered activity. How these are achieved is specified in suitable extension of CODAM. Further extensions are then made to include emotional values of stimuli. All of these extensions are supported by recent experimental brain imaging data on various working memory tasks, which are simulated with reasonable accuracy. We conclude our analysis by a discussion on the nature of cognition as seen in terms of the resulting extended attention model framework. PMID- 17081729 TI - Extraocular muscle quantification using mathematical morphology: a semi-automatic method for analyzing muscle enlargement in orbital diseases. AB - Mathematical morphology was used to quantify the dimensions of the human extraocular muscles in computed tomography orbital scans. Coronal images of 28 patients with Graves orbitopathy and 5 controls (218 orbital scans) were analyzed with an algorithm for extraocular muscle segmentation. The results showed that measurements of extraocular muscle area obtained with semi-automatic segmentation are highly correlated with manual tracing and provides a simple method to quantify orbital structures including the extraocular muscles. PMID- 17081730 TI - Second-generation peptidomimetic inhibitors of antigen presentation effectively treat autoimmune diseases in HLA-DR-transgenic mouse models. AB - Peptidomimetic compounds that bind to major histocompatibility complex class II molecules and are resistant to cathepsins can competitively inhibit the presentation of processed protein antigens. Therefore, compounds that bind to autoimmune disease-associated class II molecules are expected to compete with autoantigens for presentation and thereby interrupt the disease process. The first generation of such competitors developed for rheumatoid arthritis associated HLA-DR molecules, although resistant to cathepsins, has remained sensitive to plasma proteases, and was thus unlikely to be effective in vivo. We have therefore produced a second generation of compounds that are resistant to cathepsins and stable in plasma while maintaining binding affinity for HLA-DR molecules associated with rheumatoid arthritis and multiple sclerosis. Selected compounds of this series are shown to inhibit antigen presentation in vivo, as well as effectively treat collagen induced arthritis and experimental autoimmune encephalomyelitis in HLA-DR transgenic mouse models. PMID- 17081731 TI - The peptide specificities of the autoantibodies elicited by mouse hepatitis virus A59. AB - Synthetic decapeptides (N=206) covering the entire sequence of mouse liver fumarylacetoacetate hydrolase (FAH) were used to analyze the specificities of the autoantibodies (autoAb) elicited towards this enzyme in mice infected with mouse hepatitis virus (MHV). These autoAb bound mainly to N- and C-terminal FAH peptides, the most reactive sequences being 1-50 and 390-420, respectively. Surprisingly, although FAH sequence 1-50 shares a high degree of homology with various MHV proteins, the C-terminal portion does not. Moreover, whereas the autoAb reacted with homologous peptides surrounding residues 70, 160 and 360, non similar sequences around residues 130, 210, 240, 250, and 300 were also recognized, indicating that autoAb were not restricted to epitopes with sequence homologies. There was also a lack of correlation between the amount of anti-MHV or anti-FAH antibodies produced and the reactivity towards the peptides. Moreover, the spectrum of peptides recognized by the autoAb of a given mouse did not change significantly with time, which suggests that the MHV-elicited autoimmune response does not induce an epitope recognition spreading. Finally, anti-FAH Ab produced after immunization with rat liver FAH recognized essentially the same mouse FAH regions than autoAb from MHV-infected mice. Results indicated that the induction of the autoAb is not only related to molecular or structural mimicry, but rather supports the Danger model, in which any aggression, in this case the MHV infection, is susceptible to trigger the production of autoAb. PMID- 17081733 TI - Human papillomavirus is commonly present in psoriatic skin and normal skin samples from healthy subjects. PMID- 17081732 TI - Patients with atherosclerotic syndrome, negative in anti-cardiolipin assays, make IgA autoantibodies that preferentially target domain 4 of beta2-GPI. AB - Autoantibodies targeting beta2-glycoprotein l (beta2-GPI), a component of the atherosclerotic plaque, are commonly found in patients with acute ischemic syndromes. Serum samples from APS (antiphospholipid syndrome) patients and from cardiovascular patients exhibiting acute atherosclerotic syndromes were analyzed for IgG and IgA antibodies in both anti-beta2-GPI and anticardiolipin (aCL) ELISA assays. All of the APS samples used here were positive in both assays. Serum samples from 382 atherosclerosis patients were also analyzed for IgG and IgA antibodies in the same assays. In sharp contrast to the APS samples, we found that only 1% of the samples from atherosclerosis patients were positive for IgA aCL, and 1.6% positive for IgG aCL, whereas 35.6% were positive for IgA anti beta2-GPI and only 1.6% for IgG anti-beta2-GPI. The antigenic specificity of 29 serum samples from atherosclerosis patients was evaluated. Six different recombinant domain-deleted mutants (DM) of human beta2-GPI and full-length human beta2-GPI (wild-type) were used in competitive inhibition assays to inhibit the autoantibodies from binding in the anti-beta2-GPI ELISA assays. Domain-deleted mutants D--345 and D--45 inhibited the binding in the IgA anti-beta2-GPI assay, suggesting that these autoantibodies recognize domain 4 of the beta2-GPI molecule. These results clearly show that IgA anti-beta2-GPI autoantibodies from atherosclerotic patients are distinct from IgA autoantibodies found in APS samples. PMID- 17081734 TI - Tbx16 cooperates with Wnt11 in assembling the zebrafish organizer. AB - The organizer, the signaling center that specifies vertebrate axial polarity and the nervous system, is a dorsal midline mesodermal domain in the gastrula that will form prechordal plate and anterior notochord. We show that in zebrafish the organizer is not a single domain when it first arises in the nascent mesoderm at the onset of gastrulation. Rather, in the presumptive prechordal plate region, the organizer is subdivided into two side-by-side cellular fields. Within minutes, concurrent medial and anterior cellular movements merge, or 'coalesce', the two fields to form the well-known singular midline field. Coalescence forms a symmetrical domain because the cell movements on the left and right sides initiate simultaneously and occur synchronously. However, in embryos with reduced function of the T-box transcription factor Tbx16 (Spadetail) or its genetic target paraxial protocadherin (Papc), synchrony is lost, coalesence is disrupted, and the midline domain is misshaped. Furthermore, with combined loss of Tbx16 and Wnt11 (Silberblick), coalesence is essentially absent. Possibly as a consequence, both the anterior movement of presumptive prechordal plate and organizer function, as assayed by eye-field separation, are disrupted. Our findings thus reveal that Tbx16, in combination with Wnt11, are critical components not only in morphogenesis but also in initial assembly of the organizer. PMID- 17081735 TI - Language models based on Hebbian cell assemblies. AB - This paper demonstrates how associative neural networks as standard models for Hebbian cell assemblies can be extended to implement language processes in large scale brain simulations. To this end the classical auto- and hetero-associative paradigms of attractor nets and synfire chains (SFCs) are combined and complemented by conditioned associations as a third principle which allows for the implementation of complex graph-like transition structures between assemblies. We show example simulations of a multiple area network for object naming, which categorises objects in a visual hierarchy and generates different specific syntactic motor sequences ("words") in response. The formation of cell assemblies due to ongoing plasticity in a multiple area network for word learning is studied afterwards. Simulations show how assemblies can form by means of percolating activity across auditory and motor-related language areas, a process supported by rhythmic, synchronized propagating waves through the network. Simulations further reproduce differences in own EEG&MEG experiments between responses to word- versus non-word stimuli in human subjects. PMID- 17081736 TI - Temporal representation and reasoning in medicine: Research directions and challenges. AB - OBJECTIVE: The main aim of this paper is to propose and discuss promising directions of research in the field of temporal representation and reasoning in medicine, taking into account the recent scientific literature and challenging issues of current interest as viewed from the different research perspectives of the authors of the paper. BACKGROUND: Temporal representation and reasoning in medicine is a well-known field of research in the medical as well as computer science community. It encompasses several topics, such as summarizing data from temporal clinical databases, reasoning on temporal clinical data for therapeutic assessments, and modeling uncertainty in clinical knowledge and data. It is also related to several medical tasks, such as monitoring intensive care patients, providing treatments for chronic patients, as well as planning and scheduling clinical routine activities within complex healthcare organizations. METHODOLOGY: The authors jointly identified significant research areas based on their importance as for temporal representation and reasoning issues; the subjects were considered to be promising topics of future activity. Every subject was addressed in detail by one or two authors and then discussed with the entire team to achieve a consensus about future fields of research. RESULTS: We identified and focused on four research areas, namely (i) fuzzy logic, time, and medicine, (ii) temporal reasoning and data mining, (iii) health information systems, business processes, and time, and (iv) temporal clinical databases. For every area, we first highlighted a few basic notions that would permit any reader--including those who are unfamiliar with the topic--to understand the main goals. We then discuss interesting and promising directions of research, taking into account the recent literature and underlining the yet unresolved medical/clinical issues that deserve further scientific investigation. The considered research areas are by no means disjointed, because they share common theoretical and methodological features. Moreover, subjects of imminent interest in medicine are represented in many of the fields considered. CONCLUSIONS: We propose and discuss promising subjects of future research that deserve investigation to develop software systems that will properly manage the multifaceted temporal aspects of information and knowledge encountered by physicians during their clinical work. As the subjects of research have resulted from merging the different perspectives of the authors involved in this study, we hope the paper will succeed in stimulating discussion and multidisciplinary work in the described fields of research. PMID- 17081737 TI - Estradiol sustained release from high affinity cyclodextrin hydrogels. AB - Hydrogels for loading estradiol and controlling its release were prepared cross linking various cyclodextrins with ethyleneglycol diglycidylether. To select the more adequate cyclodextrins, estradiol solubility diagrams in water with beta cyclodextrin (betaCD), methyl-beta-cyclodextrin (MbetaCD), hydroxypropyl-beta cyclodextrin (HPbetaCD), and sulfobutyl-beta-cyclodextrin (SBbetaCD) were made in absence and presence of hydroxypropyl methylcellulose (HPMC) applying or not autoclaving. Although all cyclodextrins showed enough complexation capability, the low solubility of betaCD and the high anionic character of SBbetaCD hindered the cross-linking process, and these cyclodextrins were discarded for preparing hydrogels. Hydrogels prepared with MbetaCD (20%, 25%) or HPbetaCD (20%, 25%, and 30%), with or without HPMC 0.25%, absorbed 4-10 times their weight in water and loaded up to 24 mg estradiol per gram, which is 500 times greater than the amount of drug that can be dissolved in their aqueous phase. Positive linear correlation was found between the stability constant and the network/water partition coefficients of drug. The hydrogels sustained the release up to one week; the affinity of estradiol for the cyclodextrin units controlling the process, as shown by the negative correlation with the release rate constants. These results highlight the potential of cyclodextrin complexation for the development of hydrogels useful in loading hydrophobic drugs and controlling their release. PMID- 17081738 TI - Maternal dietary n-3 fatty acids alter immune cell fatty acid composition and leukotriene production in growing chicks. AB - The effect of feeding different amounts of n-6 and n-3 fatty acids (FA) to hens on immune tissue FA composition and leukotriene production of hatched chicks was investigated. Hens were fed diets supplemented with either 3.0% sunflower oil (Diet I), 1.5% sunflower+1.5% fish oil (Diet II), or 3.0% fish oil (Diet III) for 46 days. The hatched chicks were fed a diet containing C18:3n-3, but devoid of longer chain n-6 and n-3 FA, for 21 days. Spleen docosahexaenoic acid (DHA) content was higher in chicks from hens fed Diet III (P<0.05). The bursa content of arachidonic acid was lower in chicks hatched from hens fed Diet III (P<0.05), and the ratio of n-6 to n-3 FA was significantly higher in bursa of chicks hatched to hens fed Diet I (P<0.05). Eicosapentaenoic acid (EPA) and DHA contents were higher in bursa of chicks hatched from hens fed Diet III (P<0.05). Thrombocytes from chicks hatched to hens fed Diet III produced the most leukotriene B(5) (LTB(5)). The ratio of LTB(5) to LTB(4) concentrations was also highest (P<0.05) in chicks hatched to hens fed Diet III. These results indicate that modulating maternal dietary n-6 and n-3 FA may alter leukotriene production in chicks, which could lead to less inflammatory-related disorders in poultry. PMID- 17081739 TI - Molecular phylogeny of Coniochaetales. AB - Although the taxonomy of ascomycetes has changed dramatically, generic delimitation within the recently proposed order Coniochaetales has not been resolved. In order to clarify the phylogenetic relationships of genera in the Coniochaetaceae, we performed a molecular study based on the analyses of the sequences of the partial SSU and of the variable domains of the LSU rDNA genes. The phylogenetic trees obtained do not support the monophyly of the genera Coniochaeta, Coniochaetidium, Ephemeroascus, and Poroconiochaeta. A morphological study confirmed that there were not enough differences to distinguish these genera, and the latter three are treated as synonyms of Coniochaeta. The phialidic anamorph proved to be an informative phylogenetic character in Coniochaetales, while that the type of ascomata (cleistothecial or perithecial) and the ornamentation of the ascospore walls were of little taxonomic value at the generic level. The circumscription of the genus Coniochaeta is revised. The genera Coniocessia and Coniolariella are proposed as new within the order Xylariales to accommodate Coniochaeta nodulisporioides, and C. gamsii, respectively. The taxonomic position of Synaptospora and Wallrothiella subiculosa are also discussed. PMID- 17081740 TI - Diverse tulasnelloid fungi form mycorrhizas with epiphytic orchids in an Andean cloud forest. AB - The mycorrhizal state of epiphytic orchids has been controversially discussed, and the state and mycobionts of the pleurothallid orchids, occurring abundantly and with a high number of species on stems of trees in the Andean cloud forest, were unknown. Root samples of 77 adult individuals of the epiphytic orchids Stelis hallii, S. superbiens, S. concinna and Pleurothallis lilijae were collected in a tropical mountain rainforest of southern Ecuador. Ultrastructural evidence of symbiotic interaction was combined with molecular sequencing of fungi directly from the mycorrhizas and isolation of mycobionts. Ultrastructural analyses displayed vital orchid mycorrhizas formed by fungi with an imperforate parenthesome and cell wall slime bodies typical for the genus Tulasnella. Three different Tulasnella isolates were obtained in pure culture. Phylogenetic analysis of nuclear rDNA sequences from coding regions of the ribosomal large subunit (nucLSU) and the 5.8S subunit, including parts of the internal transcribed spacers, obtained directly from the roots and from the fungal isolates, yielded seven distinct Tulasnella clades. Tulasnella mycobionts in Stelis concinna were restricted to two Tulasnella sequence types while the other orchids were associated with up to six Tulasnella sequence types. All Tulasnella sequences are new to science and distinct from known sequences of mycobionts of terrestrial orchids. The results indicate that tulasnelloid fungi, adapted to the conditions on tree stems, might be important for orchid growth and maintenance in the Andean cloud forest. PMID- 17081741 TI - Solidification and recycling of incinerator bottom ash through the addition of colloidal silica (SiO2) solution. AB - The possibility of using incinerator bottom ash as a substitute for natural aggregates was investigated. Rough, porous surface of bottom ash, which diminishes the strength of solidified products, was improved by colloidal silica solution. As a result, a significant increase of mechanical strength was accomplished by a slight amount of silica (<1 wt% to total). Moreover, pozzolanic reaction was induced in initial cement hydration due to the nano-particle size of about 20 nm in colloidal silica solution. Cylindrical specimens and bricks were prepared from bottom ash added to a colloidal silica (SiO2) solution and cement, and then their compressive strengths were evaluated. Cylindrical specimens showed an increase of approximately 60% in compressive strength when colloidal solution containing 4 wt% silica particles was sprayed onto the bottom ash. The strength of bricks containing colloidal silica was in excess of 20 MPa, which meets the requirement of construction materials. Results of leaching tests based on Toxicity Characteristic Leaching Procedure (TCLP) proved that the solidified bottom ash possessed good chemical stability. PMID- 17081742 TI - A hydroquinone biosensor using modified core-shell magnetic nanoparticles supported on carbon paste electrode. AB - A hydroquinone biosensor was developed and used to determine hydroquinone concentration in compost extracts based on the immobilization of laccase on the surface of modified magnetic core-shell (Fe(3)O(4)-SiO2) nanoparticles. Laccase was covalently immobilized on the magnetic nanoparticles by glutaraldehyde, which was modified with amino groups on its surface. The obtained magnetic bio nanoparticles were attached to the surface of carbon paste electrode with the aid of a permanent magnet to determine hydroquinone. A good microenvironment for retaining the bioactivity of laccase was provided by the immobilization matrix. The linear range for hydroquinone determination was 1 x 10(-7) to 1.375 x 10( 4)M, with a detection limit of 1.5 x 10(-8)M. The current reached 95% of the steady-state current within about 60s. Hydroquinone concentration in compost extracts was determined by laccase biosensor and HPLC, the results of the two methods were approximately the same. PMID- 17081743 TI - Bioelectrocatalytic detection of theophylline at theophylline oxidase electrodes. AB - Bioelectrocatalytic oxidation of theophylline was studied at gold and graphite electrodes modified with microbial theophylline oxidase (ThOx), a multi-cofactor redox enzyme capable of selective oxidation of theophylline. Gold electrodes were additionally modified with self-assembled monolayers (SAMs) of (-OH)- and ( NH(2))-terminated alkanethiols of different chain lengths, to achieve compatibility between ThOx and the electrode surface. On graphite, ThOx was either physically co-adsorbed with a surfactant didodecyldimethylammonium bromide (DDAB), or entrapped within an Os-redox-polymer film. At all electrodes, ThOx was bioelectrocatalytically active; direct electrochemistry of ThOx in the absence of theophylline was followed only at the SAM-modified gold electrodes. Direct electrochemistry of ThOx correlated with redox transformations of the heme domain of ThOx, with a E(o/)of -110+/-2 mV versus Ag|AgCl, at pH 7. Bioelectrocatalytic oxidation of theophylline was optimal at mixed (-OH)/(-NH(2))-terminated SAMs; co adsorption of ThOx with DDAB improved the bioelectrocatalytic performance of the ThOx-electrode. In both cases, the response to theophylline was within the mM range. Alternatively, a reagentless ThOx-electrode based on ThOx cross-linked within the Os-redox-polymer matrix demonstrated a linear response to theophylline within the physiologically important 0.02-0.6mM (3.6-72 mg l(-1)) concentration range with a sensitivity of 52.1+/-7.8 mA cm(-2)M(-1). PMID- 17081744 TI - The EORTC QLQ-HDC29: a supplementary module assessing the quality of life during and after high-dose chemotherapy and stem cell transplantation. AB - High-dose chemotherapy followed by haematopoietic stem cell transplantation can be associated with high physical and emotional distress levels and reduced quality of life. Systematic prospective measurement of impact of therapy on patient quality of life can aid treatment choices and provide better patient information. We describe the development of a high-dose chemotherapy questionnaire module to supplement the European Organisation for Research and Treatment of Cancer Core Questionnaire (EORTC QLQ-C30). Phases 1-3 of module development were conducted in United Kingdom, Germany, Austria and Norway, according to EORTC QOL Group guidelines. Forty-eight quality of life (QOL) issues were generated from the literature searches and interviews with health care professionals (n=24) and patients (n=92). This produced a 50 item provisional module. Further testing in 169 patients resulted in the QLQ-HDC29 module, containing 29 items, conceptualised into six multi-item scales and eight single items. The EORTC QLQ-C30, supplemented by QLQ-HDC29 will provide a comprehensive QOL measure for the international clinical trials of high-dose chemotherapy. PMID- 17081746 TI - Evidence for the functional role of residues in the B'-C loop of baboon cytochrome P450 side-chain cleavage (CYP11A1) obtained by site-directed mutagenesis, kinetic analysis and homology modelling. AB - To gain further insight into the structure/function relationship of cytochrome P450 side-chain cleavage (CYP11A1), this enzyme was investigated in the Cape baboon (Papio ursinus). Four constructs were cloned and characterised in non steroidogenic mammalian COS-1 cells. Wild type recombinant baboon CYP11A1 cDNA yielded a K(m) value of 1.6 microM for 25-hydroxycholesterol. The single amino acid substitutions, I98Q and I98K resulted in a 1.7- and 2.8-fold increases in K(m) values, respectively. Conversely, the introduction of the mutation, K103A, resulted in a 1.8-fold decrease in K(m). A homology model of CYP11A1, based on the crystal structures of CYP102 and CYP2C5, revealed that residues 98 and 103 lie within the B'-C loop and contribute to the spatial orientation and structural integrity of this domain. Based on these results we propose a topological model of the CYP11A1 active pocket, which is supported by substrate docking analysis and kinetic studies. PMID- 17081745 TI - Urinary phytoestrogen excretion of rats bearing methylnitrosourea-induced mammary carcinoma in response to treatment with 2-methoxyestradiol. AB - The effect of treating mammary tumor-bearing rats with 2-methoxyestradiol (2 MeE2) on the urinary excretion of 12 phytoestrogens was investigated and compared with the changes in urinary excretion of estradiol metabolites. Alterations of excretion were registered for isoflavonoids, lignans and coumestans. However, due to large variations statistical significant differences were found only for two lignans, i.e. significant increases of enterodiol and matairesinol. Since the single components of phytoestrogens showed diverse alterations, excretions were expressed also by the ratio of total isoflavonoids to total lignans and compared with the estrogen ratios 2-hydroxyestrone to 16alpha-hydroxyestrone and A-ring to D-ring metabolites. The ratio of isoflavonoids to lignans was consistently decreased, whereas both ratios of estradiol metabolites were highly increased. The latter effect is probably due to demethylation of 2-methoxyestrone resulting in high catechol estrogen levels in urine. These results suggest that the high levels of catechol estrogens, produced by 2-MeE2 treatment, may have influenced the urinary excretion pattern of phytoestrogens. PMID- 17081747 TI - Influence of soil-water ratio on the performance of slurry phase bioreactor treating herbicide contaminated soil. AB - The influence of soil-water ratio was studied on the performance of the slurry phase bioreactor operated in sequencing batch mode (anoxic-aerobic-anoxic microenvironments) during the bioremediation of soil contaminated with pendimethalin. The performance of the reactors was evaluated at different soil water ratios (1:5-1:25; at soil loading rate (60 kg of soil/cum-day to 12 kg of soil/cum-day)) keeping the loading rate of pendimethalin constant (133.2 g/kg of soil-day) in six reactors and variable (66.6 g/kg of soil-day to 166.6 g/kg of soil-day) in other four reactors. At 1:20 soil-water ratio, the slurry phase system showed enhanced degradation of substrate (629 microg pendimethalin/g soil). The removal efficiency of pendimethalin in the reactors was dependent on the mass-transfer rates of the substrate from the soil to the aqueous phase. Soil water ratio and substrate loading rates showed significant influence on the substrate portioning, substrate degradation efficiency and substrate desorption rate. PMID- 17081748 TI - Enhanced production of Scenedesmus spp. (green microalgae) using a new medium containing fermented swine wastewater. AB - Fermented swine urine (3%) (v/v) was added to a control medium (CT), named KEP I, and an aquatic microalgal culture (10% Bold's Basal Medium) for growing mixed Scenedesmus species. During a two-month period, the KEP I medium effected, the delayed onset of the stationary phase of cell division in a batch culture. After 31 days, of culturing, the growth rate (3-fold), dry weight (2.6-fold) and amino acid levels (2.7-fold), and secondary metabolites including chlorophyll a (2.1 fold), astaxanthin (2.8-fold), lutein (2.7-fold) and alpha (greater than 30-fold) and beta-carotene (greater than 5-fold) increased a greater degree in Scenedesmus grown in KEP I medium than in CT medium. Total lipids were much less in cells grown in KEP I than those grown in CT. An increased quantum yield of photosystem II of the aquatic microalgae. The KEP I medium should improve the cost efficiency of industrial mass batch cultures for CO(2) sequestration, bioremediation, phytonutrients, agricultural fertilizers, and microalgal stock for the species preservation of aquaculture strains for use in young fish feed. It may also serve to attenuate negative environmental impact via the recycling of animal wastewater. PMID- 17081749 TI - Influence of temperature and pH on biomass production and protein biosynthesis in a putative Spirulina sp. AB - The influence of temperature and pH on biomass production and protein biosynthesis in a Spirulina sp. isolated from an oil-polluted brackish water environment in the Niger Delta was studied. The isolated organism was identified on the basis of its phenotypic characteristics such as nature and direction of helix, temperature, pH and salt tolerance ranges. Biomass concentration in the culture media was calculated as cell dry weight. The combination of 30 degrees C and pH 9.0 gave the highest values of 4.9 mg/ml and 48.2g/100 g for biomass and total crude protein, respectively. The effect of pH was modulated by temperature and vice versa during biomass production. This native isolate of Spirulina sp. offers a good source of natural protein that could be easily accepted by rural communities as single cell protein in the form of feed, food and health supplement when properly processed. PMID- 17081750 TI - Changes in organic - C, N, P and K and enzyme activities in vermicompost of biodegradable organic wastes under liming and microbial inoculants. AB - The aim of this work was to study the effect of different organic wastes, viz. cow dung, grass, aquatic weeds and municipal solid waste with lime and microbial inoculants on chemical and biochemical properties of vermicompost. Cow dung was the best substrate for vermicomposting. Application of lime (5 g/kg) and inoculation of microorganisms increased the nutrient content in vermicompost and also phosphatases and urease activities. Bacillus polymyxa, the free-living N fixer, increased N-content of vermicompost significantly (p < or = 0.01) as compared to other inoculants. PMID- 17081751 TI - CCR5 receptor antagonists: discovery and SAR study of guanylhydrazone derivatives. AB - High throughput screening (HTS) led to the identification of the guanylhydrazone of 2-(4-chlorobenzyloxy)-5-bromobenzaldehyde as a CCR5 receptor antagonist. Initial modifications of the guanylhydrazone series indicated that substitution of the benzyl group at the para-position was well tolerated. Substitution at the 5-position of the central phenyl ring was critical for potency. Replacement of the guanylhydrazone group led to the discovery of a novel series of CCR5 antagonists. PMID- 17081752 TI - What is a positive sentinel lymph node in a breast cancer patient? A practical approach. AB - Sentinel lymph node (SN) biopsy has become increasingly used for the staging of breast carcinoma, resulting in the upstaging of this disease, and this has led to concerns with regard to what should be considered a positive SN. Factors influencing the positive staging of an SN include metastasis size, the method used for metastasis detection, the definition of metastasis and the individual pathologist. Until evidence to the contrary emerges, an SN should be considered positive if metastases (nodal involvement >0.2mm in the largest dimension) are detected in it by histology. A target size should be identified, and SNs, as the most likely sites of nodal metastases, should be searched systematically to find (nearly) all of the targeted metastases. The European guidelines for SN assessment have set two such target sizes: as a minimum, all metastases >2mm should be identified, and optimally all micrometastases should also be sought. PMID- 17081753 TI - How relevant is breast cancer screening in the Asia/Pacific region? AB - Nationwide breast cancer screening with mammography is well-established in many western countries. Many studies have shown its benefit in mortality reduction. However, breast cancer screening in Asia has been slow to implement, as it was perceived that the breast cancer incidence was low and the parenchyma of Asian women was difficult to assess mammographically. With changing disease patterns, breast cancer is now one of the most common cancers amongst women in Asia. Breast cancer screening has become an important and relevant health problem. However, apart from biological differences between western and Asian women, cultural and economic considerations need to be addressed. This paper looks at the various issues pertaining to the feasibility and relevance of population-based screening in the Asia/Pacific region. PMID- 17081754 TI - Locally advanced breast cancer in a 15th century painting in Milan. AB - This previously unreported Renaissance painting depicts a young woman with locally advanced breast cancer reaching out towards a healing potion in the church of Santa Maria della Grazia in Milan, that houses Leonardo da Vinci's Last supper. Today's potion may well contain neoadjuvant systemic therapy including trastuzumab that may allow breast conserving surgery and targeted intraoperative radiotherapy. PMID- 17081755 TI - Scrib controls Cdc42 localization and activity to promote cell polarization during astrocyte migration. AB - BACKGROUND: Mammalian Scribble (Scrib) plays a conserved role in polarization of epithelial and neuronal cells. Polarization is essential for migration of a variety of cell types; however, the function of Scrib in this context remains unclear. Scrib has been shown to interact with betaPIX, a guanine nucleotide exchange factor for the small GTPases Rac and Cdc42. Cdc42 controls cell polarity from yeast to mammals during asymmetric cell division and epithelial cell polarization, as well as during cell migration. Cdc42 is, in particular, required for polarization and orientation of astrocytes in a scratch-induced polarized migration assay. Using this assay, we characterized Scrib function during polarized cell migration. RESULTS: Depletion of Scrib by siRNA or expression of dominant-negative constructs inhibits astrocyte polarization. Like Cdc42, Scrib controls protrusion formation, cytoskeleton polarization, and centrosome and Golgi reorientation. Scrib interacts and colocalizes with betaPIX at the front edge of polarizing astrocytes. Perturbation of Scrib localization or of Scrib betaPIX interaction inhibits betaPIX polarized recruitment. We further show that betaPIX is required for astrocyte polarization and that both the Scrib-binding motif and the GEF activity of betaPIX are essential for its function. Scrib and betaPIX control Cdc42 activation and localization during astrocyte polarization. Thereby, Scrib regulates Cdc42-dependent APC and Dlg1 recruitment to the leading edge to promote cell orientation. CONCLUSION: We conclude that Scrib plays a key role in the establishment of cell polarity during migration. By interacting with betaPIX, Scrib controls localization and activation of the small GTPase Cdc42 and regulates Cdc42-dependent polarization pathways. PMID- 17081756 TI - Gait initiation in children with cerebral palsy. AB - The task of gait initiation (GI), or taking a first step from a static standing position requires the development of forward momentum while maintaining dynamic balance as the body's center of mass (COM) moves forward and outside the base of support. The dynamics of GI in children (aged 7-12 years) with hemiplegic cerebral palsy (CP), diplegic CP and children with typical development (TD) were compared to characterize the mechanics and control of this task. Ground reaction forces (GRFs) and muscle activity were collected during GI at three different self-selected speeds (slow, moderate and fast). Movement of a sacral marker was also tracked to estimate downward shifting of the body during the GI task. Results demonstrate the presence of a motor sequence characterized by increased forward momentum development with increased GI speed for all groups of children. Anticipatory movements of children with CP were different when compared to children with TD. Children with hemiplegic CP demonstrated decreased lateral shifting while children with diplegic CP demonstrated a trend of decreased downward shifting of the body compared to children of TD. Analysis of the GI motor sequence in children provides a means to characterize coordination and motor control of a functional ambulatory task in children with CP compared to children with TD. PMID- 17081757 TI - Patellofemoral disorders presenting as bilateral locked knees in achondroplasia. AB - Patellofemoral disorders in achondroplasia, to our knowledge, have not been reported in the literature. We describe an unusual case of bilateral locked knees in an achondroplastic patient secondary to intra-articular subluxating patellae and symmetrical chondral flap lesions of the femoral trochlea which were treated by arthroscopic resection. We suggest that abnormal kinematics may cause patellofemoral complications in achondroplasia. PMID- 17081758 TI - QSAR study of anticoccidial activity for diverse chemical compounds: prediction and experimental assay of trans-2-(2-nitrovinyl)furan. AB - In this work we report a QSAR model that discriminates between chemically heterogeneous classes of anticoccidial and non-anticoccidial compounds. For this purpose we used the Markovian Chemicals in silico Design (MARCH-INSIDE) approach J. Mol. Mod.2002, 8, 237-245; J. Mol. Mod.2003, 9, 395-407]. Linear discriminant analysis allowed us to fit the discriminant function. This function correctly classifies 86.67% of anticoccidial compounds and 96.23% of inactive compounds in the training series. Overall classification is 94.12%. We validated the model by means of an external predicting series, with 86.96% of global predictability. Remarkably, the present model is based on topological as well as configuration dependent molecular descriptors. Therefore, the model performs timely calculations and allows discrimination between Z/E and chiral isomers. Finally, to exemplify the use of the model in practice we report the prediction and experimental assay of trans-2-(2-nitrovinyl)furan. It is notable that lesion control was 72.86% at mg/kg of body weight with respect to 60% at 125 mg/kg for amprolium (control drug). The back-projection map for this compound predicts a high level of importance for the double bond and for the nitro group in the trans position. We conclude that the MARCH-INSIDE approach enables the accurate fast track identification of anticoccidial hits. Moreover, trans-2-(2-nitrovinyl)furan seems to be a promising drug for the treatment of coccidiosis. PMID- 17081759 TI - Hybrid molecules between distamycin A and active moieties of antitumor agents. AB - The DNA minor groove is an attractive target for the design and development of molecules able to specifically recognize predetermined DNA sequences. The pyrrole amide skeleton of distamycin A has been also used as DNA sequence selective vehicle for the delivery of alkylating functions to DNA targets. Selectivity for specific sequences may be of particular importance in affecting the activity of regulatory genes (oncogenes and tumor suppressor genes). Recent work on a number of hybrid compounds, in which known antitumor compounds or simple active moieties of known antitumor agents have been tethered to distamycin frame or hairpin polyamides derived from distamycin, is reviewed. The DNA alkylating and growth inhibition activities against several tumor cell lines are reported and discussed in terms of their structural differences in relation to both the number of N methyl pyrrolic rings and the type of the alkylating unit tethered to the oligopyrrolic frame. PMID- 17081760 TI - Solid-phase synthesis of a combinatorial library of dihydroceramide analogues and its activity in human alveolar epithelial cells. AB - Solid-phase synthesis of a small combinatorial library of dihydroceramide analogues as mixtures of erythro and threo diastereomers is described. Some dihydroceramide analogues cause growth arrest and apoptosis in a dose-dependent manner in human alveolar epithelial cells. This activity is likely due to the threo isomers, as evidenced by cellular studies with a pair of diastereomerically pure N-acyldihydrosphingosines. The apoptotic activity reported in this work provides information for the design of new compounds that may provide the basis for the generation of biochemical tools for the study of different pathologies where ceramide and/or dihydroceramide are involved. PMID- 17081761 TI - Discovery and synthesis of new immunosuppressive alkaloids from the stem of Fissistigma oldhamii (Hemsl.) Merr. AB - Three new alkaloids (1-3) and twenty-one known compounds were isolated from the stem of Fissistigma oldhamii (Hemsl.) Merr. which was the ruler herb in an approved Traditional Chinese herbal formula used for treatment of rheumatoid arthritis in China and synthesis of one new immunosuppressive alkaloid was achieved. These compounds, including the crude extracts of this herb, exhibited strong activities in the inhibition of T and B cell proliferation. PMID- 17081762 TI - Semi-empirical systematics of (n,3He) reaction cross-section at 14.6 and 20 MeV. AB - A new semi-empirical formula for the evaluation of (n,(3)He) reaction cross sections at the energy of 14.6 and 20 MeV is presented. Formula was derived using the analytical expression for the evaluation of the (3)He spectrum within the frame of pre-equilibrium exciton model. The systematics obtained is compared with the empirical formula for the (n,(3)He) reaction cross-section. PMID- 17081763 TI - Synthesis of 3H-labeled N-(3-iodoprop-2E-enyl)-2beta-carbomethoxy-3beta-(4 methylphenyl)nortropane (PE2I) and its interaction with mice striatal membrane fragments. AB - PE2I is a high-affinity dopamine transporter (DA(T)) ligand that exhibits high selectivity for the DA(T) over the serotonin (5-HT(T)) and norephinephrine transporter (NE(T)) making its carbon-11 and iodine-125 labeled forms very useful for quantification of DA(T) binding sites in vivo. In this paper, we reported the synthesis of tritium-labeled PE2I, as well as improvements in the preparation of its acid precursor and PE2I itself with significant improvement in the total yield compared to previously published results. The new radioligand was evaluated as a tool for assessing DA(T) binding sites in vitro. The radioligand binding to mice striatal membranes demonstrated a homogeneous population of DA(T) binding sites with the K(d) value 9 nM. PMID- 17081764 TI - Effect of passive cardiac containment on ventricular synchrony and cardiac function in awake dogs. AB - OBJECTIVE: Passive restraint of the left ventricle (LV) has been shown to have beneficial effects on acute hemodynamics and reverse remodeling in both animal and human models. The goals of this study were to test whether a left ventricular support device (LVSD) improves LV synchrony and/or affects cardiac performance. METHODS: Ten dogs were chronically instrumented to measure hemodynamics and LV volume (sonomicrometry). Congestive heart failure (CHF) was induced by repeated intracoronary microembolization via a chronically implanted coronary catheter. The LVSD was implanted after establishment of CHF in five animals, and five animals were observed as controls. All animals were then observed for 8 weeks. A mathematical model to measure LV synchrony was used to evaluate LV motion over time. RESULTS: Mean arterial pressure and LV pressures was significantly increased after LVSD therapy, and LV pressure-volume relationships were shifted leftwards, although no change was seen in ejection fraction, end-systolic elastance, or LV dP/dt versus control. There was no significant change in diastolic function in LVSD animals compared with control animals. End-diastolic volumes were reduced by 15% after 8 weeks with LVSD treatment, versus an increase of 8% in control animals (p<0.05). Synchrony was significantly improved with LVSD therapy compared with control (9% vs 76% of baseline) in 1 of 11 ventricular dimension axes (Anterior-Apex). CONCLUSIONS: LVSD therapy provided only minimal improvement in ventricular synchrony and partially improved hemodynamics. Further study into mechanisms of benefit are warranted. PMID- 17081765 TI - Dysphagia due to tuberculosis. AB - OBJECTIVE: Dysphagia due to tuberculosis is rare in both the developing countries with high prevalence rates and the western population following the recent upsurge linked to the AIDS and immigration. AIM: To study tuberculosis as an aetiological factor in the causation of dysphagia and to evaluate the outcome of anti-tubercular treatment and surgical results in these patients. METHODS: Retrospective review of experience with 14 cases of dysphagia due to tuberculosis encountered between 1996 and 2003. RESULTS: The duration of symptoms ranged between 3 and 18 months. All of them underwent oesophagogastroscopy, barium swallow, fiberoptic bronchoscopy and CT scan of the chest. The aetiology was subcarinal node enlargement in seven, tracheo-oesophageal fistula in four, oesophageal ulcer in two and cervical node suppuration in one. Tuberculous involvement was confirmed by pathological examination in all patients. All of them received anti-tuberculous therapy. Seven patients required surgery, transthoracic repair of tracheo-oesophageal fistula in four patients, one patient required subcarinal node excision and two needed abscess drainage. There were no mortalities and there was complete relief of dysphagia in all of them. CONCLUSIONS: Tuberculosis as a causative factor for dysphagia should be considered in regions with high incidences of tuberculosis and in immunocompromised patients. Treatment with anti-tuberculous therapy is effective. Surgery is required only for complications of tuberculosis. PMID- 17081766 TI - A biophysical model of atrial fibrillation to define the appropriate ablation pattern in modified maze. AB - OBJECTIVE: The surgical Maze III procedure remains the gold standard in treating atrial fibrillation (AF); however due to clinical difficulties and higher risks, less invasive ablation alternatives are clinically investigated. The present study aims to define more efficient ablation patterns of the modified maze procedure using a biophysical model of human atria with chronic AF. METHODS: A three-dimensional model of human atria was developed using both MRI-imaging and a one-layer cellular model reproducing experimentally observed atrial cellular properties. Sustained AF could be induced by a burst-pacing protocol. Ablation lines were implemented in rendering the cardiac cells non-conductive, mimicking transmural lines. Lines were progressively implemented respectively around pulmonary veins (PV), left atrial appendage (LAA), left atrial isthmus (LAI), cavo-tricuspid isthmus (CTI), and intercaval lines (SIVC) in the computer model, defining the following patterns: P1=PV, P2=P1+LAA, P3=P2+LAI, P4=P3+CTI, P5=P3+SIVC, P6=P5+CTI. Forty simulations were done for each pattern and proportion of sinus rhythm (SR) conversion and time-to-AF termination (TAFT) were assessed. RESULTS: The most efficient patterns are P5, P6, and Maze III with 100% success. The main difference is expressed in decreasing mean TAFT with a correlation coefficient R=-0.8. There is an inflexion point for 100% success rate at a 7.5s TAFT, meaning that no additional line is mandatory beyond pattern P5. CONCLUSIONS: Our biophysical model suggests that Maze III could be simplified in his right atrial pattern to a single line joining both vena cavae. This has to be confirmed in clinical settings. PMID- 17081767 TI - Mitral valve periprosthetic leakage: Anatomical observations in 135 patients from a multicentre study. AB - OBJECTIVE: Prosthetic valve dysfunction after mitral valve replacement (MVR) may be caused by several factors, which often lead to repeated surgery. One of the most frequent determinants of reoperation is periprosthetic leakage (PPL). A few published reports have analysed PPL incidence and postoperative results after MVR, but no specific attention has been paid towards the potential relation between anatomical factors and PPL occurrence, particularly not bacterial related. The aim of this study was to evaluate the location of PPL after MVR through a multicentre retrospective study. METHODS: Between January 1985 and November 2005, 135 patients underwent reoperation at four institutions because of PPL after MVR and met the study inclusion criteria. The mitral valve annulus (MVA) was analysed in a clockwise format, indicating 12 o'clock as the mid-point of anterior annulus as viewed from the atrium. RESULTS: Overall hospital mortality was 3.7% (five patients). Repair of PPL was carried out in 83 cases whereas prosthetic valve replacement was necessary in 52 cases. The total number of sectors involved in PPL was 244. PPL occurred more frequently between hour 5 and hour 6, and hour 10 and hour 11, with the risk of leakage being, 2.8 and 2.0 times higher, respectively, than in any other portion of the MVA. CONCLUSIONS: Our study suggests that PPL occurs more frequently at antero-lateral and postero medial segments of MVA. This finding might be linked to unusual anatomical and functional factors of the MVA and may call for adjunctive care to these sectors of MVA when performing suture placement during MVR. PMID- 17081768 TI - Splenectomy changes the pattern of cytokine production in beta-thalassemic patients. AB - A major cause of morbidity and mortality in beta-thalassemic patients is infections, assumed to be the result of immunological changes. To determine the possible defect, we investigated the cytokine productions by blood cells of beta thalassemic patients using in-vivo and in-vitro methods. Heparinized blood samples collected aseptically from 22 beta-thalassemic children aged 10-12yrs (half of them were splenectomized). Samples from 10 healthy children served as control group. Part of samples was used for evaluation of plasma IL-2, IL-10 and TGF-beta1. Other part were stimulated with a mixture of LPS and PHA (1 and 10 microg/ml final concentration), for different time period (4, 24, 48 and 72h). Results showed circulating TGF-beta1 of splenectomized patients was significantly higher (p<0.01) than the control group. In-vitro results showed IL-2 production of patients' groups were significantly (p<0.01) lower than corresponding value obtained for the control group. In addition, IL-10 production by splenectomized group were less than other two group (p<0.01), while their TGF-beta1 were higher (p<0.001) at all time points treated. In conclusion, multi-transfusions could be responsible for a change in the subset of circulating lymphocytes that could contribute to a state of partial immune deficiency in beta-thalassemic patients, which is more prominence among the splenectomized patient. PMID- 17081769 TI - Thioredoxin and protein kinases in redox signaling. AB - Reactive oxygen species (ROS) play critical roles for the determination of cell fate by eliciting a wide variety of cellular responses, such as proliferation, differentiation and apoptosis. Many intracellular signaling pathways involved in such ROS-induced cellular responses are regulated by the intracellular redox state, which depends on the balance between the levels of oxidizing and reducing equivalents. Recently, increasing attention has been paid to the roles of thioredoxin (Trx) as a signaling intermediate beyond its intrinsic antioxidant activity. Especially, Trx participates in the control of the mitogen-activated protein kinase (MAPK) cascades through the redox state-dependent association and dissociation with apoptosis signal-regulating kinase 1 (ASK1), an upstream regulator of the cascades. This review highlights the current understanding of prototypical molecular mechanisms by which the redox signal is converted into the signaling through ROS-responsive protein kinases, with a special focus on the ASK1-Trx system. Understanding of such mechanisms may provide the basis for therapeutic interventions in redox-related diseases including various types of cancer. PMID- 17081770 TI - Complex spatio-temporal dynamics of fMRI BOLD: A study of motor learning. AB - Many studies have investigated the temporal properties of BOLD signal responses to task performance in regions of interest, often noting significant departures from the conventionally modelled response shape, and significant variation between regions. However, these investigations are rarely extended across the whole brain nor incorporated into the routine analysis of fMRI studies. As a result, little is known about the range of response shapes generated in the brain by common paradigms. The present study finds such temporal dynamics can be complex. We made a detailed investigation of BOLD signal responses across the whole brain during a two minute motor-sequence task, and tracked changes due to learning. The multi-component OSORU (Onset, Sustained, Offset, Ramp, Undershoot) linear model, developed by Harms and Melcher (J.Neurophysiology, 2003), was extended to characterise responses. In many regions, signal transients persisted for over thirty seconds, with large signal spikes at onset often followed by a dip in signal below the final sustained level of activation. Training altered certain features of the response shape, suggesting that different features of the response may reflect different aspects of neuro-vascular dynamics. Unmodelled, this may give rise to inconsistent results across paradigms of varying task durations. Few of the observed effects have been thoroughly addressed in physiological models of the BOLD response. The complex, extended dynamics generated by this simple, often employed task, suggests characterisation and modelling of temporal aspects of BOLD responses needs to be carried out routinely, informing experimental design and analysis, and physiological modelling. PMID- 17081771 TI - Neural correlates of metaphor processing in schizophrenia. AB - OBJECTIVE: A main feature of schizophrenic thought and language disturbance is concretism, the inability to understand the figurative meaning of proverbs and metaphors. Although this is routinely tested during clinical interview, its neural basis is unknown. METHOD: We investigated processing of metaphoric sentences with functional magnetic resonance imaging (fMRI) in 12 patients with schizophrenia and 12 control subjects. Stimuli consisted of 60 novel short sentences with either metaphoric or literal meaning presented visually, intersparsed by a low level baseline (grey background). Subjects read these sentences silently and judged by button press whether they had a positive or negative connotation. RESULTS: Reading metaphors in contrast to literal sentences revealed signal changes in the left inferior frontal gyrus in the control subjects (BA 45/47) and an area 3 cm dorsal to that in the patients (BA 45). Only activation in this area was negatively correlated with the severity of concretism rated with the PANSS. Comparison between groups for the contrast metaphors vs. low level baseline revealed stronger signal changes in the control group in the right superior/middle temporal gyrus (BA 39) and the left inferior frontal gyrus (BA 45) in the patients. CONCLUSIONS: The results in the control subjects are in line with studies showing an involvement of the left inferior frontal and right lateral temporal cortex during context processing. Failure to recruit these areas in the patients may underlie schizophrenic concretism. PMID- 17081772 TI - A molecular phylogeny for the large African orchid genus Disa. AB - Phylogenetic relationships were inferred for the African subtribe Disinae (Orchidoideae, Orchidaceae), which include the large genus Disa and the small genus Schizodium. One nuclear (ITS) gene region and two plastid (trnLF and matK) gene regions were sequenced for 136 ingroup, representing 70% of all known Disinae species, as well as for 7 outgroup taxa. The combined data matrix contained 4094 characters and was analysed using parsimony and Bayesian inference. Our results show that the generic status of Schizodium can no longer be supported, as it is deeply embedded within the genus Disa. Furthermore, the currently recognised subgenera do not reflect the phylogenetic relationships and should be rejected. Several of the currently recognised sections are monophyletic, others contain misplaced elements, while some are polyphyletic. Morphological divergence, rather than convergence, has hampered previous attempts at a phylogenetic classification of the Disinae. On the basis of our molecular phylogenetic hypothesis, we propose a monotypic subtribe Disinae and a subdivision of the genus Disa into 18 sections. PMID- 17081773 TI - Contributions of rpb2 and tef1 to the phylogeny of mushrooms and allies (Basidiomycota, Fungi). AB - A phylogeny of the fungal phylum Basidiomycota is presented based on a survey of 160 taxa and five nuclear genes. Two genes, rpb2, and tef1, are presented in detail. The rpb2 gene is more variable than tef1 and recovers well-supported clades at shallow and deep taxonomic levels. The tef1 gene recovers some deep and ordinal-level relationships but with greater branch support from nucleotides compared to amino acids. Intron placement is dynamic in tef1, often lineage specific, and diagnostic for many clades. Introns are fewer in rpb2 and tend to be highly conserved by position. When both protein-coding loci are combined with sequences of nuclear ribosomal RNA genes, 18 inclusive clades of Basidiomycota are strongly supported by Bayesian posterior probabilities and 16 by parsimony bootstrapping. These numbers are greater than produced by single genes and combined ribosomal RNA gene regions. Combination of nrDNA with amino acid sequences, or exons with third codon positions removed, produces strong measures of support, particularly for deep internodes of Basidiomycota, which have been difficult to resolve with confidence using nrDNA data alone. This study produces strong boostrap support and significant posterior probabilities for the first time for the following monophyletic groups: (1) Ustilaginomycetes plus Hymenomycetes, (2) an inclusive cluster of hymenochaetoid, corticioid, polyporoid, Thelephorales, russuloid, athelioid, Boletales, and euagarics clades, (3) Thelephorales plus the polyporoid clade, (4) the polyporoid clade, and (5) the cantharelloid clade. Strong support is also recovered for the basal position of the Dacrymycetales in the Hymenomycetidae and paraphyly of the Exobasidiomycetidae. PMID- 17081774 TI - The Pleistocene history of the sheepshead minnow (Cyprinodon variegatus): Non equilibrium evolutionary dynamics within a diversifying species complex. AB - The sheepshead minnow, Cyprinodon variegatus, is a widespread fish species that typically inhabits coastal tidal marsh and mangrove swamp environments, ranging from Cape Cod, Massaschusetts to northern Mexico and into the Caribbean. This wide range crosses several biogeographic boundaries which are coincident with genetic structuring within numerous species originating in the Pleistocene. In addition, the more northerly reaches of this species range have been further subject to the evolutionary consequences of Pleistocene glaciation due to local extinction and recolonization of formerly glaciated sites. C. variegatus thus provides an excellent vertebrate model system within which to test the extent of genetic differentiation among populations in a dominant coastal ecosystem and examine patterns of historical demography in populations distributed along a latitudinal gradient. Using mitochondrial control region and ND2 sequence data, we discovered monophyletic clades within C. variegatus with divergence times within the Pleistocene, and very low gene flow between most sites. Intraspecific genetic breaks appear to correspond broadly to biogeographic or oceanic boundaries. Pleistocene climate change appears to have had dramatic impacts on the size and distribution of populations within and near the glacial margins, but has also affected populations far from formerly glaciated regions. PMID- 17081776 TI - Eye closure sensitivity and epileptic syndromes: A retrospective study of 26 adult cases. AB - PURPOSE: The transient, mainly generalized, together with brief changes in EEG baseline immediately after eye closure, is called 'eye closure sensitivity' (ECS) which was first reported by Robinson in 1930 and there have been limited number of studies investigating ECS and epilepsy syndromes. Therefore, we aimed to reveal the possible relationship between ECS and the epilepsy syndromes in our adult patients. PATIENTS AND METHODS: Adult patients monitored in Hacettepe University Hospitals EEG Laboratory, from January 1995 to December 2005, were screened retrospectively for the presence of ECS. During EEG recording, all patients were asked to open their eyes for at least for 10s and close their eyes at the end of this period in six different montages. Hyperventilation and photic stimulation were performed according to the standard protocol of IPS. RESULTS: Twenty-six patients have had ECS during EEG recording. Nineteen (73%) were women, and mean age of the patients was 24 years. The mean ages were 12.3+/-5.4 years for seizure onset and 18.5+/-4.9 years for initial detection of ECS. Eleven had a history of febrile seizures. There was a family history for epilepsy in five patients. Photosensitivity in their EEG was noticed in 11 patients. ECS was established in five epilepsy syndromes: eyelid myoclonia with absences (EMA); (n: 6), juvenile myoclonic epilepsy (JME); (n: 6), idiopathic generalized epilepsy (IGE with tonic clonic seizure); (n: 4), juvenile absence epilepsy (JAE); (n: 1) and idiopathic occipital lobe epilepsy (IOE) (n: 9). All patients were seizure free with or without anti-epileptic medication during follow up. CONCLUSION: ECS is more common in females. It may overlap with photosensitivity but be independent from photosensitivity. It may be seen in different epilepsy syndromes including IOE which was not reported previously. PMID- 17081775 TI - The phylogeny of the social Anelosimus spiders (Araneae: Theridiidae) inferred from six molecular loci and morphology. AB - We use fragments of three nuclear genes (Histone 3, 18SrDNA, and 28SrDNA) and three mitochondrial genes (16SrDNA, ND1, and COI) totalling approximately 4.5kb, in addition to morphological data, to estimate the phylogenetic relationships among Anelosimus spiders, well known for their sociality. The analysis includes 67 individuals representing 23 of the 53 currently recognized Anelosimus species and all species groups previously recognized by morphological evidence. We analyse the data using Bayesian, maximum likelihood, and parsimony methods, considering the genes individually as well as combined (mitochondrial, nuclear, and both combined) in addition to a 'total evidence' analysis including morphology. Most of the data partitions are congruent in agreeing on several fundamental aspects of the phylogeny, and the combined molecular data yield a tree broadly similar to an existing morphological hypothesis. We argue that such congruence among data partitions is an important indicator of support that may go undetected by standard robustness estimators. Our results strongly support Anelosimus monophyly, and the monophyly of the recently revised American 'eximius lineage', although slightly altered by excluding A. pacificus. There was consistent support for the scattering of American Anelosimus species in three clades suggesting intercontinental dispersal. Several recently described species are reconstructed as monophyletic, supporting taxonomic decisions based on morphology and behaviour in this taxonomically difficult group. Corroborating previous results from morphology, the molecular data suggest that social species are scattered across the genus and thus that sociality has evolved multiple times, a significant finding for exploring the causes and consequences of social evolution in this group of organisms. PMID- 17081777 TI - Calmodulin is essential for angiogenesis in response to hypoxic stress in endothelial cells. AB - Angiogenesis, the formation of new blood vessels that is regulated by hypoxia, is a critical process for the growth and spread of tumors. Multiple phases of this process, including migration, adhesion, and formation of new capillary tubes, are needed for optimal tumor growth. Here, a new regulatory function for Ca2+-CaM in the vascular endothelium is described. Ca2+-CaM activation induced by hypoxia in endothelial cells is essential for angiogenic cellular responses. Inhibition of Ca2+-CaM activity suppressed endothelial cell migration, adhesion on collagen I substrate, invasion and impaired in vitro endothelial cell differentiation into tube-like structures. We also reported that CaM is co-distributed with the actin structures in the lamellipodia in migrating cells, whereas the actin cytoskeleton rearrangement induced by hypoxia was disrupted and HIF-1 transcriptional activity was decreased when treated with CaM antagonists into cultures. These data indicate that Ca2+-CaM activation is more closely associated with the regulation of angiogenic key events, especially in response to hypoxic stress. PMID- 17081778 TI - Expression of PTEN and Akt phosphorylation in lipopolysaccharide-treated NIH3T3 cells. AB - PTEN is a tumor suppressor gene encoding a phosphatase, and it negatively regulates cell survival mediated by the phosphoinositol 3-kinase (PI3-Kinase)-Akt pathway. To elucidate PTEN expression and its effect on the PI3-kinase-Akt pathway in fibroblasts and macrophages, we investigated the expression of PTEN and the phosphorylation status of Akt in NIH3T3 and RAW264.7 cells treated with LPS. Phosphorylation of Akt was induced by LPS treatment in a dose-dependent manner in RAW264.7 cells, but not in NIH3T3 cells. LPS induced the expression of PTEN in a dose and time-dependent manner in NIH3T3 cells (0-1 microg/ml, 0-6h). However, LPS did not stimulate PTEN expression in RAW264.7 cells. These data indicate the existence of diverse mechanisms for PTEN expression and Akt activation in fibroblasts and macrophages. RNA interference using double-stranded RNA specific for the PTEN gene reduced both mRNA and protein levels of PTEN in NIH3T3 cells treated or not with LPS. The phosphorylation status of Akt in NIH3T3 cells stimulated with LPS did not change when the PTEN expression had been inhibited by RNA interference. The present results suggest that the up-regulation of PTEN expression by LPS is not involved in the activation of Akt in NIH3T3 cells. PTEN expression might be involved in the diverse inflammatory responses to LPS in fibroblasts and macrophages. PMID- 17081779 TI - G2 checkpoint-dependent DNA repair and its response to catalase in Down syndrome and control lymphocyte cultures. AB - The amount of DNA lesions repaired in G2 and also G2 timing are controlled by the DNA damage-dependent checkpoint. Down syndrome (DS) lymphocytes showed twice as much constitutive DNA damage in G2 than control ones, when recording it as chromosomal aberrations in metaphase, after caffeine-induced checkpoint abrogation. During G2, DS lymphocytes repaired 1.5 times more DNA lesions than control ones. However the DS cells displayed a decreased threshold for checkpoint adaptation, as the spontaneous override of the G2 to mitosis transition block induced by the checkpoint took place in the DS cells when they had three times more DNA lesions than controls. Catalase addition to cultures scavenges hydrogen peroxide diffused from cells, resulting in subsequent intracellular depletion (Antunes and Cadenas, 2000). The intracellular H2O2 level seemed to regulate the G2 checkpoint. Thus, in controls, H2O2 depletion (induced by 3.2-50 microg/mL catalase) prevented its functioning: chromosomal damage increased while G2 shortened. Conversely, in the DS lymphocytes, 12.5 microg/mL catalase lengthened G2 and decreased chromosomal damage, in spite that the amount of DNA repaired in G2 was half of that repaired in the catalase-free DS lymphocytes. PMID- 17081780 TI - Characterization and expression of amphioxus ApoD gene encoding an archetype of vertebrate ApoD proteins. AB - Here we report a homologue of the apolipoprotein D gene (AmphiApoD) in amphioxus, Branchiostoma belcheri tsingtauense, the first such finding in a basal chordate cephalochordate. The main features of the protein predicted from AmphiApoD are characteristic of the apolipoprotein D. Phylogenetic analysis places AmphiApoD at the base of the phylogenetic tree, suggesting that AmphiApoD is the archetype of the vertebrate ApoD genes. Both whole mount in situ hybridization and Northern blotting and RT-PCR as well as in situ hybridization histochemistry reveal that AmphiApoD is expressed in tissues derived from mesoderm and endoderm including notochord and hind-gut, which contrasts with the strong expression patterns of ApoD genes in the ectodermal derivatives in mammals and birds. The expression profiles of the ApoD gene may have been changed to be expressed in the endo mesodermal derivatives in amphioxus after the vertebrate and cephalochordate lineages diverged; alternatively, the ApoD gene may first have been expressed in the endo-mesoderm during embryogenesis in the last common ancestor of all chordates, and subsequently came to be expressed in the ectodermal derivatives of vertebrates including mammals and birds. PMID- 17081781 TI - Jagged2-expressing hematopoietic progenitors promote regulatory T cell expansion in the periphery through notch signaling. AB - Cellular interactions promoting the in vivo expansion of CD4(+)CD25(+)Foxp3(+) regulatory T (Treg) cells for maintenance of immune tolerance remain poorly defined. Here we report that mobilized Lin(-)Sca-1(+)c-kit(+) (LSK) hematopoietic progenitor cells (HPCs), unlike medullary hematopoietic stem cells (HSCs), selectively drove the direct, immediate expansion of functional host-derived Treg cells, thereby preventing the progression to overt spontaneous autoimmune diabetes in nonobese diabetic mice. Treg cell expansion required cell-to-cell contact and Notch3 signaling, which was mediated selectively through the Notch ligand Jagged2 expressed by the multipotent HPC subset, as assessed by small interfering RNA (siRNA) silencing. Conversely, notwithstanding their similar multilineage microchimerism, neither sorted Jagged2(-) HPCs nor Jagged2(lo) medullary HSCs were able to expand Treg cells. These data provide evidence for a productive Notch-mediated interaction between a unique subset of mobilized hematopoietic progenitors and Treg cells. They open therapeutic perspectives for autologous transplantation of Jagged2(+) LSK progenitors to promote Treg cell expansion in T cell-mediated diseases. PMID- 17081782 TI - SHP1 phosphatase-dependent T cell inhibition by CEACAM1 adhesion molecule isoforms. AB - T cell activation through the T cell receptor (TCR) is subsequently modified by secondary signals that are either stimulatory or inhibitory. We show that CEACAM1 adhesion molecule isoforms containing a long cytoplasmic domain inhibited multiple T cell functions as a consequence of TCR ligation. Overexpression of CEACAM1 resulted in decreased proliferation, allogeneic reactivity, and cytokine production in vitro and delayed type hypersensitivity and inflammatory bowel disease in mouse models in vivo. Conditioned deletion of CEACAM1 in T cells caused increased TCR-CD3 complex signaling. This T cell regulation was dependent upon the presence of immunoreceptor tyrosine-based inhibition motifs (ITIM) within the cytoplasmic domain of CEACAM1 and the Src homology 2 domain-containing protein tyrosine-phosphatase 1 (SHP1) in the T cell. Thus, CEACAM1 overexpression or deletion in T cells resulted in T cell inhibition or activation, respectively, revealing a role for CEACAM1 as a class of inhibitory receptors potentially amenable to therapeutic manipulation. PMID- 17081783 TI - Negative regulation of T cell activation and autoimmunity by the transmembrane adaptor protein LAB. AB - LAB (linker for activation of B cells), also known as NTAL (non-T cell activation linker), is a LAT (linker for activation of T cells)-like adaptor protein that is expressed in B, NK, and mast cells. Its role in lymphocytes has not been clearly demonstrated. Here, we showed that aged LAB-deficient (Lat2(-/-)) mice developed an autoimmune syndrome. Lat2(-/-) T cells were hyperactivated and produced more cytokines than Lat2(+/+) T cells. Even though LAB was absent in naive T cells, LAB could be detected in activated Lat2(+/+) T cells. LAT-mediated signaling events were enhanced in Lat2(-/-) T cells; however, they were suppressed in T cells that overexpressed LAB. Mice with the Lat2 gene conditionally deleted from T cells also developed the autoimmune syndrome like Lat2(-/-) mice. Together, these data demonstrated an important role of LAB in limiting autoimmune response and exposed a mechanism regulating T cell activation. PMID- 17081784 TI - Degradation of human hemoglobin by Prevotella intermedia. AB - In this study, the ability of Prevotella intermedia, an obligate anaerobic rod, to degrade human hemoglobin was determined by SDS-PAGE and the degradation was quantified by scanning densitometry. Both bacterial cells and culture supernatants degraded hemoglobin. The hemoglobin degradation by P. intermedia was time-dependent, heat sensitive, pH related and was not influenced by iron restriction. Inhibition studies demonstrated that a cysteine protease might be involved in hemoglobin degradation and this protease might require metal ions for its activity and it might be thiol-requiring and trypsin-inducible. The results indicate that P. intermedia is capable to release heme from hemoglobin, hence provide a source of iron for its proliferation. PMID- 17081785 TI - A mutation in the gene encoding cytochrome c1 leads to a decreased ROS content and to a long-lived phenotype in the filamentous fungus Podospora anserina. AB - We present here the properties of a complex III loss-of-function mutant of the filamentous fungus Podospora anserina. The mutation corresponds to a single substitution in the second intron of the gene cyc1 encoding cytochrome c(1), leading to a splicing defect. The cyc1-1 mutant is long-lived, exhibits a defect in ascospore pigmentation, has a reduced growth rate and a reduced ROS production associated with a stabilisation of its mitochondrial DNA. We also show that increased longevity is linked with morphologically modified mitochondria and an increased number of mitochondrial genomes. Overexpression of the alternative oxidase rescues all these phenotypes and restores aging. Interestingly, the absence of complex III in this mutant is not paralleled with a deficiency in complex I activity as reported in mammals although the respiratory chain of P. anserina has recently been demonstrated to be organized according to the "respirasome" model. PMID- 17081786 TI - Molecular characterization and phylogenetic analysis of BjussuMP-I: a RGD-P-III class hemorrhagic metalloprotease from Bothrops jararacussu snake venom. AB - Snake venom metalloproteases (SVMPs) embody zinc-dependent multidomain enzymes responsible for a relevant pathophysiology in envenomation, including local and systemic hemorrhage. The molecular features responsible for hemorrhagic potency of SVMPs have been associated with their multidomains structures which can target these proteins them to several receptors of different tissues and cellular types. BjussuMP-I, a SVMP isolated from the Bothrops jararacussu venom, has been characterized as a P-III hemorrhagic metalloprotease. The complete cDNA sequence of BjussuMP-I with 1641bp encodes open reading frames of 547 amino acid residues, which conserve the common domains of P-III high molecular weight hemorrhagic metalloproteases: (i) pre-pro-peptide, (ii) metalloprotease, (iii) disintegrin like and (iv) rich cysteine domain. BjussuMP-I induced lyses in fibrin clots and inhibited collagen- and ADP-induced platelet aggregation. We are reporting, for the first time, the primary structure of an RGD-P-III class snake venom metalloprotease. A phylogenetic analysis of the BjussuMP-I metalloprotease/catalytic domain was performed to get new insights into the molecular evolution of the metalloproteases. A theoretical molecular model of this domain was built through folding recognition (threading) techniques and refined by molecular dynamics simulation. Then, the final BjussuMP-I catalytic domain model was compared to other SVMPs and Reprolysin family proteins in order to identify eventual structural differences, which could help to understand the biochemical activities of these enzymes. The presence of large hydrophobic areas and some conserved surface charge-positive residues were identified as important features of the SVMPs and other metalloproteases. PMID- 17081787 TI - Routine metabolic rate of southern bluefin tuna (Thunnus maccoyii). AB - Routine metabolic rate (RMR) was measured in fasting southern bluefin tuna, Thunnus maccoyii, the largest tuna species studied so far (body mass=19.6 kg (+/ 1.9 SE)). Mean mass-specific RMR was 460 mg kg(-1) h(-1) (+/-34.9) at a mean water temperature of 19 degrees C. When evaluated southern bluefin tuna standard metabolic rate (SMR) is added to published values of other tuna species, there is a strong allometeric relationship with body mass (423 M(0.86), R(2)=0.97). This demonstrates that tuna interspecific SMR scale with respect to body mass similar to that of other active teleosts, but is approximately 4-fold higher. However, RMR (not SMR) is most appropriate in ram-ventilating species that are physiologically unable to achieve complete rest. Respiration was measured in a large (250,000 l) flexible polypropylene respirometer (mesocosm respirometer) that was deployed within a marine-farm sea cage for 29 days. Fasted fish were maintained within the respirometer up to 42 h while dissolved oxygen dropped by 0.056 (+/-0.004) mg l(-1) h(-1). Fish showed no obvious signs of stress. They swam at 1.1 (+/-0.1) fork lengths per second and several fed within the respirometer immediately after measurements. PMID- 17081788 TI - Metabolic and genetic regulation of cardiac energy substrate preference. AB - Proper heart function relies on high efficiency of energy conversion. Mitochondrial oxygen-dependent processes transfer most of the chemical energy from metabolic substrates into ATP. Healthy myocardium uses mainly fatty acids as its major energy source, with little contribution of glucose. However, lactate, ketone bodies, amino acids or even acetate can be oxidized under certain circumstances. A complex interplay exists between various substrates responding to energy needs and substrate availability. The relative substrate concentration is the prime factor defining preference and utilization rate. Allosteric enzyme regulation and protein phosphorylation cascades, partially controlled by hormones such as insulin, modulate the concentration effect; together they provide short term adjustments of cardiac energy metabolism. The expression of metabolic machinery genes is also dynamically regulated in response to developmental and (patho)physiological conditions, leading to long-term adjustments. Specific nuclear receptor transcription factors and co-activators regulate the expression of these genes. These include peroxisome proliferator-activated receptors and their nuclear receptor co-activator, estrogen-related receptor and hypoxia inducible transcription factor 1. Increasing glucose and reducing fatty acid oxidation by metabolic regulation is already a target for effective drugs used in ischemic heart disease and heart failure. Interaction with genetic factors that control energy metabolism could provide even more powerful pharmacological tools. PMID- 17081789 TI - Does overfeeding enhance genotype effects on energy metabolism and lipid deposition in breast muscle of ducks? AB - We evaluated the effects of genotype (Muscovy, Pekin and their crossbreed hinny and mule ducks) and feeding levels (overfeeding between 12 and 14 weeks of age vs ad libitum feeding) on energy metabolism and lipid deposition in breast muscle of ducks. Samples of breast muscle (Pectoralis major) were collected at 14 weeks of age from 8 birds per group. Overfeeding induced an accumulation of lipids in breast muscle (1.5- to 1.7-fold, depending on genotype) mainly induced by triglyceride deposition. It also induced a considerable increase in the amounts (expressed as g/100 g of tissue) of saturated and mono-unsaturated fatty acids (SFA, MUFA), while the amounts of poly-unsaturated fatty acids (PUFA) remained unchanged in hinny and Muscovy ducks or slightly increased in Pekin and mule ducks. In breast muscle, overfeeding decreased the activity of the main enzymes involved in lipogenesis from glucose (glucose-6-phosphate dehydrogenase, G6PDH, malic enzyme, ME, acetyl CoA carboxylase, ACX). Lipoprotein lipase (LPL) activity in Pectoralis major muscle was also significantly decreased (-21%). The ability of muscle tissues to catabolize long-chain fatty acids, as assessed by beta hydroxyacyl CoA dehydrogenase (HAD) activity, was increased in Pectoralis major muscle, as was cytochrome-c oxidase (COX) activity. Hybrid and Pekin ducks exhibited higher levels of ACX and LPL activity in Pectoralis major muscle than Muscovy ducks, suggesting a greater ability to synthesise lipids in situ, and to take up circulating lipids. Total lipid content in breast muscle of hybrid and Pekin ducks was higher than in that of Muscovy ducks. In hybrid and Pekin ducks, lipid composition of breast muscle was characterized by higher amounts of triglycerides, SFA and MUFA than in Muscovy ducks. Finally, oxidative metabolism was greater in Pectoralis major muscles of hybrid and Pekin ducks than in Muscovy ducks, suggesting an adaptative strategy of muscle energy metabolism according to lipid level. PMID- 17081790 TI - Comparison of ACE activity in amphibian tissues: Rana esculenta and Xenopus laevis. AB - Angiotensin converting enzyme (ACE) is the dipeptidyl-carboxypeptidase of the renin-angiotensin system involved in the control of blood pressure and hydromineral metabolism. It converts angiotensin I to angiotensin II, the biologically active octapeptide. Angiotensin converting enzyme-like activity has been demonstrated in a wide range of vertebrates. The presence of ACE was investigated in tissues of two amphibian species, the frog Rana esculenta and the toad Xenopus laevis. ACE activities were determined by specific substrate hydrolysis in gut, gonads, lung, kidney, heart, liver, skin, erythrocytes, and muscle homogenates and plasma by means of high performance liquid chromatography. Significant ACE activity was found in gut, gonads, lung and kidney, while that in heart, liver, skin, erythrocytes, muscle, and plasma was very low. Testis of toad contained the highest ACE activity, while that in erythrocytes of male and female frogs was notable. PMID- 17081791 TI - Development of a qRT-PCR assay to determine the relative mRNA expression of two different trypsins in Atlantic cod (Gadus morhua). AB - A fluorescence resonance energy transfer (FRET) based quantitative RT-PCR method (qRT PCR) was developed in this study for measuring the mRNA expression of trypsins Y and I in the Atlantic cod. Atlantic cod beta-actin was used as the reference gene and standard curves were created for quantification of the mRNA expression levels. For yet unknown reasons, the Atlantic cod (Gadus morhua) produces several trypsins with different characteristics. Trypsin I is the most common and best characterized of these but trypsin Y is a recently discovered enzyme. The recombinant form of trypsin Y was found to have unique characteristics relative to trypsin I. The native form of trypsin Y has proven difficult to isolate from the cod and activity assays do not distinguish between the activities of trypsin I and trypsin Y. The results show that trypsin Y mRNA is expressed in a very low copy number relative to that of trypsin I (ratio of 1:1340), which may explain the difficulty of isolating the native form of trypsin Y. PMID- 17081792 TI - Validation of the Japanese version of the Quality of Life-Assessment of Growth Hormone Deficiency in Adults (QoL-AGHDA). AB - OBJECTIVE: To evaluate validity and reliability of the Japanese version of the Quality of Life-Assessment of Growth Hormone Deficiency in Adults (QoL-AGHDA). DESIGN: Observational study; cross-sectional, longitudinal. METHODS: Seventy-five adults with growth hormone deficiency completed the SF-36 (a generic health related QOL scale) and the QoL-AGHDA before growth hormone replacement therapy and approximately 3 weeks later (when the therapy began). A sample (n=1000) of controls from the general population was also studied. We computed rates of missing data, measured reproducibility and internal consistency reliability, and tested for known-groups validity, concurrent validity, unidimensionality (by principle component analysis), and content validity. RESULTS: Rates of missing data were low (0-1.4%). The mean of QoL-AGHDA scores in the patients was 8.2 (SD, 6.4). The scores were reproducible (k=0.41-0.78), and internally consistent (alpha=0.91) and the scale was unidimensional. QoL-AGHDA scores were associated with SF-36 scores as hypothesized. Scores were significantly higher in the patients than in controls (8.1+/-0.7, and 5.6+/-0.2, P<0.001). Discrimination between patients and controls was slightly better using scores on the "General Health" and "Role Physical" subscale of the SF-36 as explanatory variables than using QoL-AGHDA scores. CONCLUSIONS: The QoL-AGHDA's reliability, validity, and rates of missing data were satisfactory, and the scale was confirmed to be unidimensional. However, because some subscales of the SF-36 were better for discriminating patients from controls, the content validity of the QoL-AGHDA may need to be re-evaluated. PMID- 17081793 TI - [How I carry out...laparoscopic hysterectomy in large uterus]. PMID- 17081794 TI - Shape analysis of microcalcifications using Radon transform. AB - Microcalcifications are one of the early signs of breast cancer, and they are of great importance for an early diagnosis. Moreover, the spatial distribution and the shape of the microcalcifications have a significant impact in medical practice to evaluate the probability of malignancy of the tumor. In this work a method, performing computer-aided classification of the shape of calcifications accordingly to the classification scheme proposed by Le Gal, is presented. In the first stage, in order to remove mammographic background, the image is preprocessed with a matched filter, designed by modeling the microcalcifications as Gaussian spots and the image as a Fractional Brownian Motion. Afterwards, morphology of spots has been evaluated using two different sets of parameters. The first set utilizes the moments of inertia of the second and third order to compute a set of features, which are invariant to rotations and translations of the image. The second set of parameters is derived from the evaluation of the Radon transform, as computed along eight axes. The results of the Radon transform are used to associate to each lesion a set of features, which are invariant to rotation and scaling of the image. In the final stage, a multilayer neural network has been used to assign each microcalcification to the classes introduced by Le Gal. The topology of the neural network is the same for both sets of descriptors, in order to allow comparison of the discriminative power of the two feature sets. Experimental results obtained with the proposed method from a set of digitized mammograms are reported and discussed. PMID- 17081795 TI - The influences of stenosis on the downstream flow pattern in curved arteries. AB - The influence of stenosis on the pulsatile blood flow pattern in curved arteries with stenosis at inner wall was investigated by computer simulations. Numerical calculations were performed with various values of physiological parameters to examine the effect of a stenosis on the hemodynamic characteristics such as secondary flow, flow separation, wall shear stress (WSS) and pressure drop. The results demonstrated that when the severity of a stenosis at the inner wall of a curved artery reaches a certain level, the flow pattern in the downstream of the artery shows a dramatic change compared to that of a curved artery with no stenosis. According to previous studies, a flow separation occurs at the inner wall of the bend in a curved artery. The present work reports an analysis of such a flow separation area at the inner wall of the post stenosis region in curved arteries with a stenosis. In addition, another area of flow separation with low and oscillating WSS and blood pressure at the outer wall in a downstream tube was also found and investigated. The observed characteristic change of the flow downstream may suggest a formation of a new plaque at the outer wall downstream. PMID- 17081796 TI - Deep brain stimulation of globus pallidus internus for dystonia. AB - Neuromodulation is the functional modification of neural structures through the use of electrical stimulation. Its most clinically applicable use is deep brain stimulation (DBS) of basal ganglia structures in Parkinson's disease (PD) and essential tremor (ET). More recently, it has been used as a means of treating dystonic movement disorders. The main target of DBS for dystonia is the posteroventral globus pallidus internus (GPi), although the thalamus has been used as an alternate target in a minority of cases. In comparison to the effects seen in PD, the improvement in dystonic postures appear to differ in several ways -delay of clinical benefit, higher voltage requirements, and varied stimulator settings. In this review, the authors discuss the clinical characteristics, pathophysiology, microelectrode recording (MER) signatures, optimal surgical targets, programming parameters and outcomes in dystonia. PMID- 17081797 TI - Oncostatin M decreases adiponectin expression and induces dedifferentiation of adipocytes by JAK3- and MEK-dependent pathways. AB - Adiponectin, an adipokine secreted from adipocytes, plays a crucial role in the regulation of glucose and lipid metabolism. In the present study, we examine the role of the IL-6 family of cytokines in the expression of adiponectin in human adipocytes derived from human adipose tissue-derived stromal cells. Oncostatin M (OSM), but not IL-6, attenuated the expression level of adiponectin dose- and time-dependently, and the inhibitory effect of OSM on adiponectin expression was as potent as that of TNF-alpha. The OSM-induced down-regulation of adiponectin expression was correlated with the down-regulation of PPARgamma2 and lipoprotein lipase, markers for adipogenic differentiation, and depletion of intracellular lipid droplets, suggesting dedifferentiation of adipocytes in response to OSM. OSM induced phosphorylation of STAT1, and treatment of adipocytes with JAK3 inhibitor WHI-P131 or MEK inhibitor U0126, but not with JAK2 inhibitor AG490, prevented the activation of STAT1. Furthermore, the OSM-induced suppression of adiponectin expression and dedifferentiation of adipocytes were ameliorated by WHI-P131 or U0126, but not by AG490. These results suggest that OSM inhibits adiponectin expression by inducing dedifferentiation of adipocytes through signaling pathways involving JAK3 and MEK, but not JAK2. PMID- 17081798 TI - Rotamers of m-cresol cation studied by mass-analyzed threshold ionization spectroscopy. AB - We have applied two-color resonant two-photon mass-analyzed threshold ionization technique to record the vibrational spectra of the selected rotamers of m-cresol. The adiabatic ionization energies of cis and trans m-cresol are determined to be 66,933+/-5 and 67,084+/-5 cm(-1), respectively. Frequencies of the in-plane ring vibrations 6a, 1, and 9b are measured to be 528, 720, 1167 cm(-1) for the cis and 520, 698, and 1153 cm(-1) for the trans m-cresol cation. This indicates that different orientation of the OH group with respect to the CH(3) group slightly influences these ring vibrations. PMID- 17081799 TI - Fluorescent sensing of anions with acridinedione based neutral PET chemosensor. AB - Newly synthesised fluorescent chemosensor ADDTU contains the thiourea receptor connected to the acridinedione (ADD) fluorophore via a covalent bond, giving rise to a fluorophore-receptor motif. In this fluorescent chemosensor, the anion recognition takes place at the receptor site which result in the concomitant changes in the photophysical properties of a ADD fluorophore by modulation of photoinduced electron transfer (PET) process. The binding ability of these sensor with the anions F(-), Cl(-), Br(-), I(-), HSO(4)(-), ClO(4)(-), AcO(-), H(2)PO(4)(-) and BF(4)(-) (as their tetrabutylammounium salts) in acetonitrile were investigated using UV-vis, steady state and time-resolved emission techniques. ADDTU system allows for the selective fluorescent sensing of AcO(-), H(2)PO(4)(-) and F(-) over other anions in acetonitrile. PMID- 17081800 TI - An 'Honest Broker' mechanism to maintain privacy for patient care and academic medical research. AB - PURPOSE: From the Hippocratic Oath to the World Medical Association's Declaration of Geneva, physicians have sworn to protect patients' privacy. However, as systems move to more integrated architectures, protecting this medical data becomes more of a challenge. The increase in complexity of IT environments, the aggregation of data, and the desire of other entities to access this data, often 24 h/day x 7 day/week x 365 day/year, is putting serious strains on our ability to maintain its security. This problem cuts across all electronic record sources from patient care records to academic medical research records. APPROACH: In order to address this issue, we are rethinking the way we store, transmit, process, access, and federate patient data from clinical and research applications. Our groups at the University of Michigan are developing a system called the "Honest Broker" to help manage this problem. The Honest Broker will offload the burden of housing identifiable data elements of protected health information (PHI) (e.g., name and address) as well as manage data transfer between clinical and research systems. Lab results and other non-identifiable data will be stored in separate systems with either a research study ID or clinical ID number. This two-component architecture increases the burden on attackers who now need to compromise two systems, one of which is seriously hardened, in order to match health data with a patient's actual identity. CONCLUSIONS: While no security system is truly intrusion-proof, this architecture provides a high security choke point reducing the likelihood of a breach. By redesigning the method of integrating clinical care and research, we have enabled projects that would be cost prohibitive to conduct otherwise. The scalability of this mechanism is dependant on nature of the heterogenous nature of the clinical systems serving patients. PMID- 17081802 TI - Grazing-induced changes in cell wall silicification in a marine diatom. AB - In aquatic environments, diatoms (Bacillariophyceae) constitute a central group of microalgae which contribute to about 40% of the oceanic primary production. Diatoms have an absolute requirement for silicon to build-up their silicified cell wall in the form of two shells (the frustule). To date, changes in diatom cell wall silicification have been only studied in response to changes in the growth environment, with consistent increase in diatom silica content when specific growth rates decrease under nutrient or light limitations. Here, we report the first evidence for grazing-induced changes in cell wall silicification in a marine diatom. Cells grown in preconditioned media that had contained both diatoms and herbivores are significantly more silicified than diatoms grown in media that have contained diatoms alone or starved herbivores. These observations suggest that grazing-induced increase in cell wall silicification can be viewed as an adaptive reaction in habitats with variable grazing pressure, and demonstrate that silicification in diatoms is not only a constitutive mechanical protection for the cell, but also a phenotypically plastic trait modulated by grazing. In turn, our results corroborate the idea that plant-herbivore interactions, beyond grazing sensu stricto, contribute to drive ecosystem structure and biogeochemical cycles in the ocean. PMID- 17081801 TI - Phospholipase A2 and phospholipase B activities in fungi. AB - As saprophytes or disease causing microorganisms, fungi acquire nutrients from dead organic material or living host organisms. Lipids as structural components of cell membranes and storage compartments play an important role as energy-rich food source. In recent years, it also has become clear that lipids have a wide range of bioactive properties including signal transduction and cell to cell communication. Thus, it is not surprising that fungi possess a broad range of hydrolytic enzymes that attack neutral lipids and phospholipids. Especially during infection of a mammalian host, phospholipase A(2) (PLA(2)) enzymes released by fungi could play important roles not only for nutrient acquisition and tissue invasion, but for intricate modulation of the host's immune response. Sequencing of fungal genomes has revealed a wide range of genes encoding PLA(2) activities in fungi. We are just beginning to become aware of the significance these enzymes could have for the fungal cells and their interaction with the host. PMID- 17081803 TI - Heparin-mediated extracorporeal low density lipoprotein precipitation as a possible therapeutic approach in preeclampsia. AB - Preeclampsia is a pregnancy-related hypertensive disease resulting in substantial maternal and neonatal morbidity and mortality. Until today there is no satisfactory treatment to stop disease progression except immediate delivery of the fetus. Heparin-mediated extracorporeal low density lipoprotein (LDL) precipitation (H.E.L.P.) apheresis removes simultaneously circulating LDL, lipoprotein(a) [Lp(a)], fibrinogen, C-reactive protein (CRP) and various proinflammatory and procoagulatory factors. This study was to test the feasibility of H.E.L.P. apheresis in preeclamptic patients and its potential effects on blood and placental markers of preeclampsia. We applied H.E.L.P. apheresis to nine preeclamptic patients and it was well tolerated. Their gestational ages could be continued by 17.7 (3-49) more days. Eight of the nine neonates did well during their neonatal stage. One infant died of late-onset sepsis. H.E.L.P. apheresis reduced significantly circulating levels of triglycerides, total and LDL-cholesterol, Lp(a), fibrinogen, hs-CRP, TNFalpha, sVCAM-1, E-selectin, lipopolysaccharide binding protein (LBP), homocysteine and plasma viscosity. We conclude that H.E.L.P. apheresis reduced maternal circulating levels of proinflammatory and coagulatory markers and plasma viscosity without overt maternal or neonatal clinical side effects. PMID- 17081804 TI - Two-dimensional and three-dimensional transthoracic echocardiography in surgical planning for right atrial metastatic melanoma. AB - Melanoma is the most common form of cardiac metastases. Surgical excision has been shown to be an effective palliative measure. This requires detailed definition of cardiac anatomy in relation to the tumour. Two-dimensional (2D), three-dimensional (3D) transthoracic (TTE) and transoesophageal echocardiography (TEE), spiral computerised tomography (CT) and magnetic resonance imaging (MRI) have all been described in aiding surgical planning for excision of cardiac tumours. In this case report, 3D-TTE provided excellent anatomical definition for surgical planning of a large right atrial melanoma precluding the need for more invasive and expensive investigations. PMID- 17081806 TI - Natural history of patients hospitalized for management of cirrhotic ascites. AB - BACKGROUND & AIMS: Since the International Ascites Club published the diagnostic criteria of refractory ascites (RA) and hepatorenal syndrome (HRS), there have been few studies assessing the natural history of ascites. The aims of this study were to define the natural history of cirrhotic ascites and to identify prognostic factors for dilutional hyponatremia (DH), RA, HRS, and survival. METHODS: Two hundred sixty-three consecutive cirrhotic patients were followed for 40.9 +/- 2.6 months after their first significant ascites. RESULTS: During follow up 74 (28.1%) patients developed DH, 30 (11.4%) RA (diuretic-resistant in 2 cases and diuretic-intractable because of the development of diuretic-induced complications in 28 cases), and 20 (7.6%) HRS (type 1, 7; type 2, 13). The 5-year probability of DH, RA, and HRS development was 37.1%, 11.4%, and 11.4%, respectively. The probability of survival at 1 and 5 years was 85% and 56.5%, respectively. The independent predictors for survival were baseline age, baseline Child-Pugh score, and DH development. The 1-year probability of survival after developing DH, RA, and type 2 HRS was 25.6%, 31.6%, and 38.5%, respectively. In contrast, the mean survival was only 7 +/- 2 days in those patients developing type 1 HRS. CONCLUSIONS: (1) The survival of cirrhotic patients with first episode of ascites is relatively high, and it is mainly influenced by age and Child-Pugh score at the time of ascites decompensation, as well as by DH development. (2) The probability of RA and HRS development is relatively low, but they are associated with a poor prognosis. PMID- 17081805 TI - Inhibition of rainbow trout (Oncorhynchus mykiss) P450 aromatase activities in brain and ovarian microsomes by various environmental substances. AB - Aromatase, a key steroidogenic enzyme that catalyses the conversion of androgens to estrogens, represent a target for endocrine disrupting chemicals. However, little is known about the effect of pollutants on aromatase enzymes in fish. In this study, we first optimized a rainbow trout (Oncorhynchus mykiss) microsomal aromatase assay to measure the effects of 43 substances belonging to diverse chemical classes (steroidal and non steroidal aromatase inhibitors, pesticides, heavy metals, organotin compounds, dioxins, polycyclic aromatic hydrocarbons) on brain and ovarian aromatase activities in vitro. Our results showed that 12 compounds were able to inhibit brain and ovarian aromatase activities in a dose dependent manner with IC50 values ranging from the low nM to the high microM range depending on the substance: steroidal and non steroidal inhibitors of aromatase (4-hydroxyandrostenedione, androstatrienedione, aminogluthethimide), imidazole fungicides (clotrimazole, imazalil, prochloraz), triazole fungicides (difenoconazole, fenbuconazole, propiconazole, triadimenol), the pyrimidine fungicide fenarimol and methylmercury. Overall, this study demonstrates that rainbow trout brain and ovarian microsomal aromatase assay is suitable for evaluating potential aromatase inhibitors in vitro notably with respect to environmental screening. The results highlight that methylmercury and some pesticides that are currently used throughout the world, have the potential to interfere with the biosynthesis of endogenous estrogens in fish. PMID- 17081807 TI - The placebo effect for gastroenterology: tool or torment. AB - Placebo generally has a negative connotation both in clinical practice and in clinical trials. In this report the nature of placebo is explored in general and in relation to gastrointestinal diseases. The goodness of placebo is highlighted. PubMed was searched for articles and commentaries on placebo and for systematic reviews on placebo. Selected placebo-controlled trials in gastroenterology were reviewed. There are 5 key components impacting the placebo response: patient characteristics, practitioner characteristics, practitioner-patient interaction, the nature of the illness being treated, and the treatment being applied and the setting in which it is applied. There are also a number of factors that constitute the placebo response and likely all apply to some extent. These include the natural history of the condition, a regression to the mean (accounting for fluctuation in measurements over time), other time effects (ie, patient learning to cope, practitioner gaining expertise), unidentified parallel interventions (ie, change in habits or exogenous stress), the placebo effect of the administered agent being compared with placebo, and the physiologic effects of the placebo. In summary, there are positive aspects to placebo. Clinicians treating patients with gastrointestinal diseases can harness aspects of the placebo response to improve patient outcomes. Clinical trial planners can harness aspects of the placebo response to design disease-specific trials that can minimize the placebo response. PMID- 17081808 TI - pH-tunable equilibria in azocrown ethers with histidine moieties. AB - The crown ethers with electro- and photoactive azo moieties containing substituents with mobile protons such as in the -COOH groups of histidine, show unique effect of pH switched on/off presence of the azo form. The differences observed for the electrochemical behavior of azocrown ethers with N acetylhistidine and imidazole moieties reveal the interference of a chemical reduction pathway in strongly acidified solutions. This chemical reduction process leads to the formation of a hydrazine derivative which can be detected by its further electroreduction on the electrode surface. The involvement of chemical reduction is seen clearly in the presence of mobile protons of the -COOH group and mercury as the electrode substrate. The behaviour of the N acetylhistidine azomacrocyle is similar to that of compounds known to exist in quinone-hydrazone tautomeric equilibria. PMID- 17081809 TI - Gaseous metabolism of the chicken embryo and hatchling during post-hypoxic recovery. AB - In neonatal mammals, the drop in oxygen consumption (VO2) during moderate degrees of hypoxia is a manifestation of metabolic depression, and occurs without anaerobic energy compensation. We examined the possibility that embryos also respond to hypoxia with a similar hypometabolic response, by measuring the extent of the O2 debt during post-hypoxic recovery. In chicken embryos at incubation days 11 (E11) and 16 (E16), and hatchlings on the day of hatching (H1), VO2 and carbon dioxide production (VCO2) were measured with an open flow system. The protocol consisted of 1h in air, followed by 40 min in hypoxia (either 17% or 11% O2) and additional 45 min of post-hypoxic recovery. VO2 dropped in hypoxia, more with 11% than 17% O2, and more the younger the animal. During post-hypoxic recovery VO2 returned to, but did not exceed, the pre-hypoxic level, indicating that no O2 debt was contracted during hypoxia. In H1, the changes of VCO2 during hypoxia and post-hypoxia matched those of VO2. Differently, in the embryos, the changes in VCO2 during hypoxia and post-hypoxic recovery were minimal. This phenomenon is explained by changes in the large CO2 stores of the eggs, which buffer the changes in CO2 output of aerobic origin. We conclude that in the chicken embryo and hatchling the energetic shortfall during the hypoxic decrease in VO2 is not compensated by anaerobic energy supply, and represents a phenomenon of metabolic depression. PMID- 17081810 TI - Solid-phase microextraction-liquid chromatography (SPME-LC) determination of fluoxetine and norfluoxetine in plasma using a heated liquid flow through interface. AB - A simple and sensitive procedure using solid-phase microextraction coupled with high performance liquid chromatography (HPLC) to analyze fluoxetine (FLU) and its metabolite norfluoxetine (nor-FLU) in plasma samples was developed and validated. SPME conditions were optimized employing a factorial design. The sampling step was performed using a PDMS-DVB fiber and desorption was carried out in a novel homemade heated interface. Fluoxetine and norfluoxetine were analyzed by HPLC, using a C18 Phase Sep column (150mmx4.6mm, 3microm) packed "in house", and acetonitrile:acetate buffer 25mmoll(-1) with triethylamine 25mmoll(-1) pH 4.6 (70:30) as the mobile phase. The developed method has shown precision, linearity, specificity, and limit of quantification (LOQ) adequate to assay fluoxetine and norfluoxetine in plasma. Furthermore, the results obtained using the homemade interface has shown an improvement in the desorption process when compared with the results obtained using the off-line mode. PMID- 17081811 TI - Statistics for proteomics: experimental design and 2-DE differential analysis. AB - Proteomics relies on the separation of complex protein mixtures using bidimensional electrophoresis. This approach is largely used to detect the expression variations of proteins prepared from two or more samples. Recently, attention was drawn on the reliability of the results published in literature. Among the critical points identified were experimental design, differential analysis and the problem of missing data, all problems where statistics can be of help. Using examples and terms understandable by biologists, we describe how a collaboration between biologists and statisticians can improve reliability of results and confidence in conclusions. PMID- 17081812 TI - An isocratic liquid chromatography method for determining HIV non-nucleoside reverse transcriptase inhibitor and protease inhibitor concentrations in human plasma. AB - An efficient, isocratic high performance liquid chromatography (HPLC) method for determining human immunodeficiency virus (HIV) non-nucleoside reverse transcriptase inhibitors (NNRTIs) and protease inhibitors (PIs) in plasma is advantageous for laboratories participating in clinical trials and therapeutic drug monitoring (TDM) programs, or conducting small animal research. The combination of isocratic reversed phase chromatography using an S-3, 3.0 mm x 150 mm column along with low plasma volume (200 microl), rapid liquid-liquid extraction, and detection at a single wavelength (212 nm) over a short run time makes this method valuable. Within and between assay variability ranges from 0.8 to 3.5% and 1.2-6.2%, respectively. Accuracy ranges from 91.0 to 112.8% for four quality controls (50, 100, 1000, and 10,000 ng/ml) for all drugs measured (efavirenz, nevirapine, amprenavir, atazanavir, indinavir, lopinavir, nelfinavir, ritonavir, and saquinavir). PMID- 17081813 TI - Application of HPLC to study the kinetics of a branched bi-enzyme system consisting of hypoxanthine-guanine phosphoribosyltransferase and xanthine oxidase -an important biochemical system to evaluate the efficiency of the anticancer drug 6-mercaptopurine in ALL cell line. AB - The thiopurine antimetabolite 6-mercaptopurine (6MP) is an important chemotherapeutic drug in the conventional treatment of childhood acute lymphoblastic leukemia (ALL). 6MP is mainly catabolized by both hypoxanthine guanine phosphoribosyltransferase (HGPRT) and xanthine oxidase (XOD) to form thioinosinic monophosphate (TIMP) (therapeutically active metabolite) and 6 thiouric acid (6TUA) (inactive metabolite), respectively. The activity of both the enzymes varies among ALL patients governing the active and the inactive metabolite profile within the immature lymphocytes. Therefore, an attempt was made to study the kinetic nature of the branched bi-enzyme system acting on 6MP and to quantitate TIMP and 6TUA formed when the two enzymes are present in equal and variable ratios. The quantification of the branched kinetics using spectrophotometric method presents problem due to the closely apposed lambda(max) of the substrates and products. Hence, employing an HPLC method, the quantification of the products was done with the progress of time. The limit of quantification (LOQ) of substrate was found to be 10nM and for products as 50 nM. The limit of detection (LOD) was found to be 1 nM for the substrate and the products. The method exhibited linearity in the range of 0.01-100 microM for 6MP and 0.05-100 microM for both 6TUA and TIMP. The amount of TIMP formed was higher than that of 6TUA in the bi-enzyme system when both the enzymes were present in equivalent enzymatic ratio. It was further found that enzymatic ratios play an important role in determining the amounts of TIMP and 6TUA. This method was further validated using actively growing T-ALL cell line (Jurkat) to study the branched kinetics, wherein it was observed that treatment of 50 microM 6MP led to the generation of 12 microM TIMP and 0.8 microM 6TUA in 6 h at 37 degrees C. PMID- 17081814 TI - Phenotypic variability of a 4q34-->qter inherited deletion: MRKH syndrome in the daughter, cardiac defect and Fallopian tube cancer in the mother. AB - Terminal deletions of the long arm of chromosome 4 are associated with a recognizable phenotype consisting of dysmorphic facial features, cleft palate, upper and lower limb malformations, cardiac defects and growth and mental retardation. Here we report on two female patients, a mother and her daughter, carrying the same 4q34-->qter deletion but presenting with a different phenotype. The mother's presentation is consistent with previous findings in patients with terminal deletions of the long arm of chromosome 4. However, she presented at the age of 54years with bilateral serous carcinoma of the Fallopian tubes, a rare gynaecologic cancer that might be attributed to the haploinsufficiency of the tumor suppressor gene FAT. The daughter presented isolated congenital aplasia of the uterus and vagina, the prime feature of the MRKH syndrome. This has not been described before in association with a 46,XX,del(4)(q34qter). PMID- 17081815 TI - Disassociation of bone resorption and formation by GLP-2: a 14-day study in healthy postmenopausal women. AB - We have previously shown that a single subcutaneous injection of glucagon-like peptide-2 (GLP-2) at 10 p.m. in postmenopausal women results in a dose-dependent decrease in the nocturnal serum and urine concentrations of fragments derived from the degradation of the C-terminal telopeptide region of collagen type I (s CTX and u-CTX) and u-DPD, markers of bone resorption. In contrast, bone formation, as assessed by serum osteocalcin and procollagen type I N-terminal propeptide (PINP), appeared to be unaffected by treatment with exogenous GLP-2. These effects were further investigated in a 14-day study. The aim was to demonstrate that a parenteral formulation of GLP-2 is safe and well tolerated after repeated dosing in healthy postmenopausal women for 14 days. It was further investigated whether the effects on bone turnover markers were sustained throughout the study period. The study was a double-blind placebo-controlled trial with 60 postmenopausal women and 2 different doses of GLP-2 (1.6 mg and 3.2 mg GLP-2) against a saline control. The data for bone resorption revealed a similar reduction on Day 1 and Day 14, both based on time course and AUC. There were no signs of tachyphylaxis and no serious adverse reaction. Both GLP-2 doses resulted in similar and significant (p<0.001) reduction in bone resorption indicating that the maximum efficacious dose has been approached. Osteocalcin and PINP levels were unaffected at Day 1 and Day 14, suggesting a disassociation between bone resorption and bone formation during GLP-2 treatment. PMID- 17081817 TI - Help prevent exposure to secondhand smoke. PMID- 17081818 TI - Beyond Washington: the impact of state laws and regulations on dietetics practice. PMID- 17081819 TI - Polycystic ovarian syndrome: what it is and why registered dietitians need to know. PMID- 17081820 TI - Food Allergen Labeling and Consumer Protection Act of 2004 in effect. PMID- 17081821 TI - The dos and don'ts of PowerPoint presentations. PMID- 17081822 TI - Partnerships: why ADA must choose wisely. PMID- 17081823 TI - The settling problem in calcium-fortified soybean drinks. PMID- 17081825 TI - Back to basics: have milk with meals. PMID- 17081826 TI - Meeting adequate intake for dietary calcium without dairy foods in adolescents aged 9 to 18 years (National Health and Nutrition Examination Survey 2001-2002). AB - BACKGROUND: In the United States, >50% of dietary calcium is provided by milk and milk products. Calcium intakes in the United States are inadequate for many children, and a large proportion do not drink milk or consume dairy products. However, no studies have addressed whether dairy-free diets can provide adequate calcium while meeting other nutrient recommendations. OBJECTIVE: To determine the highest calcium intake for adolescents obtained from dairy-free diets, and to examine the relationship between intakes of calcium-fortified foods, using citrus juice as an example, and maximal calcium intakes. DESIGN: In the National Health and Nutrition Examination Survey 2001 to 2002, 65 females and 62 males, aged 9 to 18 years, reported no intake of dairy. We used linear programming to generate diets with maximal calcium intake, while meeting Dietary Reference Intakes for a set of nutrients, limiting energy and fat intakes, and not selecting food quantities exceeding amounts usually eaten in the population. RESULTS: With food use and energy and fat constraints, diets formulated by linear programming provided 1,150 and 1,411 mg/day of calcium for girls and boys, respectively. With the Dietary Reference Intakes constraints, these decreased to 869 and 1,160 mg/day. When we introduced 1.5 servings of fortified juice to the diets, the highest calcium intake increased to 1,302 mg/day for girls and to 1,640 mg/day for boys. CONCLUSIONS: Adequate intake for calcium cannot be met with dairy-free diets while meeting other nutrient recommendations. To meet the adequate intake for calcium without large changes in dietary patterns, calcium-fortified foods are needed. In addition, greater physical activity and responsible sunlight exposure should be encouraged to promote vitamin D adequacy. PMID- 17081827 TI - Predictors of nutritional adequacy in mother-toddler dyads from rural families with limited incomes. AB - OBJECTIVE: To predict the mean adequacy ratio (MAR) scores of mothers and toddlers from intakes of fruits, vegetables, and dairy group foods and being seated during mealtimes. DESIGN/SUBJECTS: This was a regression analysis of cross sectional data of the diet quality and being seated during mealtimes of 100 rural mother-toddler dyads from limited-income families using two 24-hour dietary recalls. Children were 11 to 25 months of age and at or below 100% of the poverty index. MEASURES OF OUTCOME: Dietary quality for mothers and toddlers was assessed using a MAR score for eight different nutrients (vitamin A, vitamin C, vitamin D, folate, calcium, zinc, iron, and magnesium), and a score of 85 or above was considered nutritionally adequate. The main food groups of interest were servings from the fruits, vegetables, and dairy group foods. Mealtime sitting behavior was the percentage of times the toddler remained seated while eating. RESULTS: Servings of fruits, vegetables, and dairy foods predicted 0.62 of the variance in the mother's MAR score, whereas vegetable and dairy intakes along with being seated while eating indicated nutritional adequacy for toddlers. Mothers with low MAR scores were most likely to have toddlers with poor diets, although few toddlers had poor diet quality. CONCLUSIONS: Adequate intakes of dairy, vegetables, and whole fruits along with being seated while eating could be quick assessment tools to screen toddlers for nutritional risk. Mothers with poor diet quality were likely to have toddlers with poor diets; low intakes of fruits, vegetables, and dairy foods were markers for poor diet quality in mothers. PMID- 17081828 TI - Influence of nutrition attitudes and motivators for eating on postpartum weight status in low-income new mothers. AB - OBJECTIVE: The purpose of this study was to identify attitudes about nutrition and their influence on weight status in low-income mothers in the first year postpartum. DESIGN: Nutrition attitudes were assessed at 1.5, 6, and 12 months postpartum. Body weight was measured at each time point and height at 1.5 months to calculate body mass index. Nutrition attitudes at each time were compared with demographic variables and weight status. SUBJECTS: Subjects were 340 non-Hispanic white (31.3%), non-Hispanic black (25.1%), and Hispanic (43.7%) new mothers (mean age=22.4 years) located in central Texas. Criteria for participation included good health at delivery and low income (/=3 servings of dairy per day; these girls also reported higher energy intake but had lower body mass index z scores and body fat than the girls who consumed fewer than three dairy servings each day. Among plausible reporters, no relationship between dairy intake and weight status was noted. This discrepancy may be attributable to a high percentage (45%) of overweight underreporters in the total sample. Our findings reveal that reporting bias, resulting from the presence of a substantial proportion of underreporters of higher weight status, can contribute to obtaining spurious associations between dairy intake and weight status. These findings underscore the need for randomly controlled trials to assess the role of dairy in weight management. PMID- 17081837 TI - Beliefs about whole-grain foods by food and nutrition professionals, health club members, and special supplemental nutrition program for women, infants, and children participants/State fair attendees. AB - Whole-grain foods are important components of healthful diets that may help prevent chronic diseases. Consumer beliefs that influence consumption of whole grains are poorly understood. This analysis surveyed three groups regarding their beliefs about whole-grain foods. The groups were food and nutrition professionals (n=103), health club members (n=103), and individuals representing various consumer segments of the general population, including participants in the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) and state fair attendees (n=68). Most respondents were aware of the term whole-grain foods, but less often reported that they use the term. Bread and cereal were most often named as examples of whole-grain foods. Lack of processing and use of the entire grain were the major reasons a food was perceived as being a whole-grain food. The major benefit of eating whole grains was reported to be fiber intake. Food and nutrition professionals provided more differentiated responses, whereas WIC/state fair participants had fewer and less elaborate responses. Assessing beliefs about whole grains offers insights to nutrition professionals for encouraging healthful food consumption. PMID- 17081838 TI - Do Mexican-American mothers' food-related parenting practices influence their children's weight and dietary intake? AB - Food-related parenting attitudes are thought to influence children's dietary intake and weight. The objective of this study was to examine the associations between mothers' reports of food-related parenting and children's dietary intake and body mass index (BMI). A sample of 108 Mexican-American fifth-grade children and their mothers were surveyed. Children's height, weight, and three 24-hour dietary recalls were collected. Mothers reported household food insecurity status and food-related parenting attitudes. Correlational analyses were calculated among dietary intake variables, children's BMI percentiles, and food-parenting behaviors. Mothers' pressure on their children to eat was inversely correlated with children's BMI. In food-insecure families, attitudes toward making healthful foods available were inversely associated with children's daily energy intake and BMI. In contrast, in food-secure families, attitudes about making healthful foods available were positively associated with children's fruit intake and percentage energy from fat, and parental modeling of healthful food behaviors was inversely associated with the energy density. In our sample of Mexican-American families, mothers' food-related parenting was associated with their children's weight and dietary intake. These associations differed in food-secure and food-insecure households. Overall, pressure to eat was highly associated with children's weight, but the temporal nature of these relationships cannot be discerned. PMID- 17081839 TI - Effects of energy-content labels and motivational posters on sales of sugar sweetened beverages: stimulating sales of diet drinks among adults study. AB - This study examined the effects of an environmental intervention promoting more non-energy-containing beverage consumption compared to sugar-sweetened soft drink consumption through vending machines in an urban college setting. Eight soft drink vending machines were randomly selected and assigned to one of three conditions over a 9-week period: energy-content labels on non-energy-containing beverage selection panels (intervention I), labels plus motivational posters (intervention II), or control. The totals of all beverages sold and machine revenue were recorded at baseline (2 weeks), intervention (5 weeks), and postintervention (2 weeks) periods. Use of energy-content labels and motivational posters, compared with control group, resulted in a significantly lower growth rate of sugar-sweetened beverage sales (P<0.05). Total revenue for all beverages increased during the intervention period. It is estimated that the non-energy containing beverages combined accounted for 70.52% of the increased revenue. Results of this study suggest that energy-content labels and motivational posters on beverage vending machines may be an effective way to influence beverage sales. PMID- 17081840 TI - Differences in eating and activity behaviors, health history, and biomarkers among normal-weight, overweight, and obese rural midwestern hispanic women. AB - This project examined differences in health history, eating and activity behaviors, and biomarkers across three weight categories of rural, Midwestern, US born Hispanic women as a way to identify critical factors for improving long-term health. Using a cross-sectional descriptive design, a convenience sample of 70 women, aged 19 to 69 years, completed general health, demographic, food frequency, and physical activity questionnaires. Height, weight, blood pressure, and waist circumferences were measured and recorded. One-way analyses of variance and chi(2) analyses were completed. Differences in the presence of diabetes and hypertension were demonstrated across the weight categories. In all three weight categories, fat intakes exceeded and fruit, vegetable, and dairy intakes were below current recommendations. Higher mean daily grain food intake was found for women of normal weight compared to overweight women (7.1+/-5.5 vs 3.6+/-2.3 servings, respectively, P=0.01). A greater proportion of normal-weight women (61.9%) met the targeted physical activity level compared to overweight (42.9%) and obese (21.4%) women (chi(2)=8.29, P=0.016). Because differences in physical activity rather than energy intakes might be a source of energy imbalances that affect weight, additional work to identify behavioral determinants that promote adoption of healthful activity behaviors will be a key component in designing effective educational strategies for these Hispanic women. PMID- 17081841 TI - What is ADA's staffing ratio for clinical dietitians? PMID- 17081844 TI - Risk of rupture in unruptured anterior communicating artery aneurysms: meta analysis of natural history studies. AB - BACKGROUND: The decision of whether to operate on patients bearing UIAs is complicated by the limitations in current knowledge of the natural history of such lesions. The ISUIA has estimated the annual risk of rupture below that justifying surgery for most incidentally found lesions less than 7 mm in diameter. However, there is some evidence that aneurysms located in the ACoA show a higher risk of rupture, even with diameters of less than 7 mm. The present study was conducted to investigate the risk of aneurysm rupture in this site. METHODS: The available literature on the subject was thoroughly reviewed, and a meta-analysis was carried out comparing the risk of rupture of aneurysms found in the ACoA with that of aneurysms in other sites. RESULTS: Aneurysms found unruptured in the ACoA show a risk of rupture twice as high as that of other intracranial aneurysms (95% confidence interval, 1.29-3.12). It is the first time this fact has been demonstrated based on the follow-up of unruptured aneurysms. CONCLUSION: When deciding whether to operate on UIAs located in the ACoA, surgeons should consider their higher risk of rupture. PMID- 17081847 TI - Do leptomeningeal venous drainage and dysplastic venous dilation predict hemorrhage in dural arteriovenous fistula? AB - BACKGROUND: The objective of this study was to correlate the presence of leptomeningeal venous drainage and dysplastic venous dilation with the risk of intracranial hemorrhage in DAVFs. METHODS: The subjects for this research were composed of 93 patients with DAVFs who were studied retrospectively with regard to therapeutic success and failure, who had undergone either neurosurgery or embolization or a combination of both methods, and whose disease was located in the cavernous sinus, the superior sagittal sinus, or the transverse-sigmoid sinus of the anterior fossa or of the tentorium. Also among these study subjects were patients who had had angiography done in at least 6 cranial vessels (external and internal carotid and vertebral arteries) before and after treatment and who had a minimum follow-up of 3 months. Cases of leptomeningeal venous drainage and dysplastic venous dilation and their associations with intracranial hemorrhage were ascertained for all the 93 case patients included in the study. RESULTS: The presence of leptomeningeal drainage (P = .0002) and that of dysplastic venous dilation (P = .036) increased the risk of intracranial hemorrhage in case patients for DAVFs of the 5 regions. CONCLUSION: There is a statistical significance between risk of intracranial hemorrhage in DAVFs and the presence of leptomeningeal drainage and dysplastic venous dilation. PMID- 17081848 TI - Parasagittal meningiomas: follow-up review. AB - BACKGROUND: Parasagittal meningioma is one that fills the parasagittal angle, with no brain tissue between the tumor and the SSS. Invasion of the SSS is a challenge for complete removal and, consequently, for recurrence of these tumors. The objective of this study was to analyze the factors that influenced the clinical outcome of patients with parasagittal tumors surgically treated. METHODS: Review of data on 53 patients with diagnosis of parasagittal meningiomas surgically treated from 1984 to 2004. Thirty-four (64.2%) were female and 19 (35.8%) were male; age ranged from 18 to 81 years old (mean, 54.98 +/- 5.80). Follow-up ranged from 2 to 261 months (mean, 93.71 +/- 68.45). The patients were operated on using microsurgical techniques. Tumors in the anterior third (9) or occluding the SSS (5) were removed with the sinus; tumors touching/pouching the SSS (20) were removed and its dural attachment coagulated; tumors invading one sinus wall (10) were removed with partial excision and reconstruction of the sinus wall, and tumors invading more than one sinus wall in the posterior two thirds of the SSS (7) had a subtotal removal. No attempt at sinus resection and reconstruction was performed for tumors placed in the posterior two thirds of the SSS. Analysis of the patient outcome was done using survival and RFS Kaplan-Meier curves. The chi(2), Fisher exact, log-rank, Mann-Whitney, and Kruskall-Wallis ANOVA tests were used for comparing demographic data, survival curves, proportions, and medians, respectively. RESULTS: Total and subtotal resection were achieved in 85% and 13.1%, respectively. Males had better survival than females (P = .0252). Total RF rates were 10%, 25%, and 100% at 5 years and 100%, 50%, and 100% at 10 years for patients with meningiomas WHO grades I, II, and III, respectively. The RF survival curve was better for patients with grade I meningioma (grades I vs II vs III, P = .0001). There was no difference between the RF survival curves according to age, histopathologic WHO grade, location along or invasion of the SSS, and extent of resection. Males (P = .0401), WHO grade I (P < .0001), total resection (P = .0139), and less sinus invasion (P = .0308) had better RFS curves. Operative, surgery-related, and overall mortality were 1.9%, 5.4%, and 26.4%, respectively. CONCLUSIONS: Recurrence of parasagittal meningiomas predominated in males, in grades II/III tumors, after subtotal resection, and with more invasion of the SSS. Subtotal or total resections without sinus resection were considered adequate for treating these patients. PMID- 17081850 TI - Traumatic pseudoaneurysm of the middle meningeal artery and cerebral intraparenchymal hematoma: case report. AB - BACKGROUND: Pseudoaneuryms of the MMA are rare lesions, accounting for less than 1% of all intracranial aneurysms. CASE DESCRIPTION: We report a case of a spontaneous intracerebral hematoma that DSA revealed as ispilateral traumatic PMMA. The lesion was managed through an endovascular approach, and definitive embolization of pseudoaneurysm and parent vessel with histoacryl injection was carried out. Recovery and follow-up were uneventful. We discuss the management of those lesions, emphasizing the current neuroendovascular tools and techniques. CONCLUSION: Pseudoaneurysm of the MMA is uncommon but has a potential aggressive natural history. As it can occur with different patterns of intracranial hemorrhage, the external carotid arteries have to be included in hemorrhagic investigational DSA. Once diagnosed, it must be treated. The minimally invasive endovascular approach and definitive embolization of pseudoaneurysm and parent vessel with histoacryl injection are safe and effective. PMID- 17081854 TI - Cervical spinal cord compression due to an osteochondroma in hereditary multiple exostosis: case report and review of the literature. AB - BACKGROUND: Hereditary multiple exostosis is a benign disorder characterized by multiple osteochondromas affecting long and flat bones, although occasionally vertebral column involvement can be seen. Cervical spinal cord compression in HME is a rare condition. The objective of this manuscript is to describe a rare case of cervical myelopathy due to an exostosis arising from C7 in a patient with HME and a comprehensive review of the current literature. CASE DESCRIPTION: We describe a case of HME in an 18-year-old girl with myelopathy characterized by quadriparesis due to an osteochondroma arising from the lamina of C7. The patient underwent surgery, and a laminectomy was performed with a complete removal of the exostosis and spinal cord decompression. One month after surgery, patient presented an excellent recovery without neurologic deficits. CONCLUSIONS: Cervical spinal cord compression resulting from osteochondroma is an extremely serious complication of HME. Neurosurgical approach should be recommended in order to achieve a spinal cord decompression, which usually results in excellent functional recovery. PMID- 17081855 TI - Rhinosinusitis: Developing guidance for clinical trials. AB - The Rhinosinusitis Initiative was developed by 5 national societies. The current guidance document is an expansion of the 2004 publication, "Rhinosinusitis: Establishing definitions for clinical research and patient care" and provides templates for clinical trials in antimicrobial, anti-inflammatory, and symptom relieving therapies for the following: (1) acute presumed bacterial rhinosinusitis, (2) chronic rhinosinusitis (CRS) without nasal polyps, (3) CRS with nasal polyps, and (4) classic allergic fungal rhinosinusitis. In addition to the templates for clinical trials and proposed study designs, the Rhinosinusitis Initiative has developed 6 appendices, which address (1) health outcomes, (2) nasal endoscopy and staging of CRS, (3) radiologic imaging, (4) microbiology, (5) laboratory measures, and (6) biostatistical methods. PMID- 17081856 TI - Guidelines for dealing with disasters involving large numbers of extensive burns. PMID- 17081857 TI - Anterior segment imaging: new milestones, new challenges. PMID- 17081858 TI - Noninvasive corneal stromal collagen imaging using two-photon-generated second harmonic signals. AB - PURPOSE: To investigate the feasibility of using femtosecond-pulse lasers to produce second-harmonic generated (SHG) signals to noninvasively assess corneal stromal collagen organization. SETTING: The Eye Institute, University of California, Irvine, California, USA. METHODS: Mouse, rabbit, and human corneas were examined by two-photon confocal microscopy using a variable-wavelength femtosecond lasers to produce SHG signals. Two types were detected: forward scattered and backward scattered. Wavelength dependence of the SHG signal was confirmed by spectral separation using the 510 Meta (Zeiss). To verify the spatial relation between SHG signals and corneal cells, staining of cytoskeletons and nuclei was performed. RESULTS: Second-harmonic-generated signal intensity was strongest with an excitation wavelength of 800 nm for all 3 species. Second harmonic-generated forward signals showed a distinct fibrillar pattern organized into bands suggesting lamellae, while backscattered SHG signals appeared more diffuse and indistinct. Reconstruction of SHG signals showed two patterns of lamellar organization: highly interwoven in the anterior stroma and orthogonally arranged in the posterior stroma. Unique to the human cornea was the presence of transverse, sutural lamellae that inserted into Bowman's layer, suggesting an anchoring function. CONCLUSIONS: Using two-photon confocal microscopy to generate SHG signals from the corneal collagen provides a powerful new approach to noninvasively study corneal structure. Human corneas had a unique organizational pattern with sutural lamellae to provide important biomechanical support that was not present in mouse or rabbit corneas. PMID- 17081859 TI - Magnetic resonance imaging of aging, accommodating, phakic, and pseudophakic ciliary muscle diameters. AB - PURPOSE: To quantify in vivo accommodative changes in the aging human ciliary muscle diameter in phakic and pseudophakic eyes. SETTING: Department of Surgery/Bioengineering, UMDNJ-Robert Wood Johnson Medical School, Piscataway, and the Institute of Ophthalmology and Visual Science UMDNJ-New Jersey Medical School, Newark, New Jersey, USA. METHODS: Images were acquired from 48 eyes of 40 people between the ages of 22 and 91 years, 1 eye of 32 phakic volunteers and both eyes of 8 patients who had monocular implantation of a single-piece AcrySof intraocular lens (IOL) (Alcon Laboratories). Images were acquired during physiological accommodation and with accommodation at rest, and the diameter of the ciliary muscle ring was measured. RESULTS: Results show the ciliary muscle remains active throughout life. The accommodative change in its diameter (mean 0.64 mm) (P<.00001) was undiminished by age or IOL implantation. Preliminary data showed that the accommodative decrease in muscle diameter in phakic and pseudophakic eyes was statistically identical. The phakic eyes had a marked decrease in ciliary muscle diameter with advancing age for both accommodative states (P<.000001 and P<.000001), which did not appear to be altered by IOL implantation. The lens equator was constant with age in the unaccommodated human eye, resulting in decreased circumlental space with advancing age in the phakic eyes. CONCLUSION: Although the undiminished ability of the ciliary muscle to decrease its diameter with accommodation can be relied on in strategies for presbyopia correction, even in advanced presbyopia, the decreasing circumlental space and its potential effects on zonular tension must also be considered. PMID- 17081860 TI - Accuracy, repeatability, and reproducibility of Artemis very high-frequency digital ultrasound arc-scan lateral dimension measurements. AB - PURPOSE: To determine the accuracy, repeatability, and reproducibility of measurement of lateral dimensions using the Artemis (Ultralink LLC) very high frequency (VHF) digital ultrasound (US) arc scanner. SETTING: London Vision Clinic, London, United Kingdom. METHODS: A test object was measured first with a micrometer and then with the Artemis arc scanner. Five sets of 10 consecutive B scans of the test object were performed with the scanner. The test object was removed from the system between each scan set. One expert observer and one newly trained observer separately measured the lateral dimension of the test object. Two-factor analysis of variance was performed. The accuracy was calculated as the average bias of the scan set averages. The repeatability and reproducibility coefficients were calculated. The coefficient of variation (CV) was calculated for repeatability and reproducibility. RESULTS: The test object was measured to be 10.80 mm wide. The mean lateral dimension bias was 0.00 mm. The repeatability coefficient was 0.114 mm. The reproducibility coefficient was 0.026 mm. The repeatability CV was 0.38%, and the reproducibility CV was 0.09%. There was no statistically significant variation between observers (P = .0965). There was a statistically significant variation between scan sets (P = .0036) attributed to minor vertical changes in the alignment of the test object between consecutive scan sets. CONCLUSION: The Artemis VHF digital US arc scanner obtained accurate, repeatable, and reproducible measurements of lateral dimensions of the size commonly found in the anterior segment. PMID- 17081861 TI - Anterior chamber angle measurement with optical coherence tomography: intraobserver and interobserver variability. AB - PURPOSE: To assess intraobserver and interobserver variability of anterior segment optical coherence tomography (AS-OCT) as an objective diagnostic tool to quantify the anterior chamber angle and opening width. SETTING: University Eye Clinic, Lubeck, Germany. METHODS: The anterior chamber angle and opening width were assessed in 18 eyes of 9 healthy volunteers by 2 observers. Intraobserver reproducibility was evaluated by calculating an intraclass correlation coefficient (ICC) in a mixed model. Each observer had a separate model using information from 5 scans. Interobserver variability was determined by Bland Altman analysis. The ICC was calculated in a mixed model using a residual maximum likelihood method. The results of 3 and 5 repeated scans were evaluated to indicate the change to 1 measurement application. RESULTS: The mean anterior chamber angle measurement was 35.9 degrees +/- 5.7 (SD) for observer A and 36.2 +/- 5.7 degrees for observer B. The ICC was 0.94 and 0.91, respectively. The mean opening width was 315 +/- 62 microm for observer A and 317 +/- 60 microm for observer B. The ICC was 0.97 and 0.93, respectively. Interobserver comparisons showed a mean difference between anterior chamber angle measurements of -0.27 +/- 1.6 degrees, a limit of agreement (LOA) interval from -3.52 to 2.98 degrees, and an ICC estimate of 0.96. The mean difference in opening width measurements was 2.40 +/- 12.40 microm, the LOA from -27.20 to 22.40 microm, and the estimated ICC 0.96. Using 1 instead of the mean of 5 measurements, the LOA range increased by 3.46 degrees for the anterior chamber angle and 30.0 microm for the opening width. CONCLUSION: Anterior chamber angle and opening width measurements by OCT showed low intraobserver and interobserver variability, indicating OCT is a valuable technique for quantitative assessment that provides reproducible measurements and objective documentation by different examiners. PMID- 17081862 TI - Internal anterior chamber diameter using optical coherence tomography compared with white-to-white distances using automated measurements. AB - PURPOSE: To compare internal horizontal anterior chamber (AC) diameter determined by optical coherence tomography (OCT) and horizontal corneal diameter (white-to white [WTW]) using automated measurements. SETTING: Department of Ophthalmology, Johann Wolfgang Goethe-University, Frankfurt am Main, Germany. METHODS: Internal AC diameter and WTW distance was measured in 52 eyes of 26 patients using the Visante OCT (Carl Zeiss Meditec), IOLMaster (Carl Zeiss Meditec), and Orbscan IIz topography system (Bausch & Lomb). Statistical evaluation was performed using the Bland-Altman method and regression analysis for comparison of measurement techniques. The Kruskal-Wallis test was used to measure the repeatability of each device. RESULTS: The mean internal AC diameter was 12.45 mm +/- 0.53 (SD) with OCT; the mean WTW distance was 12.17 +/- 0.45 mm with the IOLMaster and 11.84 +/- 0.41 mm with the Orbscan IIz. A positive regression was determined for AC diameter and both WTW measurements. Measurement values varied little between both WTW measurement systems (R(2) = 0.9384). CONCLUSIONS: Anterior chamber measurement using optical coherence tomography (Visante OCT) was easy to handle and showed good repeatability. The internal horizontal diameter of the AC was larger than the horizontal corneal diameter determined by automated WTW measurements (IOLMaster, Orbscan IIz). Optical coherence tomography with the Visante OCT allows direct measurement of the AC width. PMID- 17081863 TI - Relationship between the corneal surface and the anterior segment of the cornea: An Asian perspective. AB - PURPOSE: To determine the values for the anterior best-fit sphere (BFS) and posterior BFS in an Asian population using the Orbscan II (Bausch & Lomb) slit scanning Placido disk corneal topographer. SETTING: Hospital-based refractive surgery center. METHODS: This prospective nonrandomized study comprised of 1 eye of 724 subjects. The eyes were measured with the Orbscan II. Default settings were used to generate relative elevation maps. Anterior BFS, posterior BFS, mean keratometry (K(mean)), spherical equivalent (SE), horizontal white-to-white size (WTW), and anterior chamber depth (ACD) were analyzed. RESULTS: The mean spherical equivalent (SE) in the 724 eyes was -5.32 diopters (D) +/- 2.88 (SD). The anterior BFS and posterior BFS were normally distributed, with a mean of 43.05 +/- 1.37 D and 52.43 +/- 2.03 D, respectively. The regression equation of anterior BFS and posterior BFS was posterior BFS = 0.443 + 1.208 anterior BFS (r(2) = 0.667, P<.01). The ratio of posterior BFS to anterior BFS was 1.22 +/- 0.03. The anterior BFS was significantly correlated to posterior BFS (P<.01), K(mean) (P<.01), and horizontal WTW (P<.01) but not to SE (P = .28). The posterior BFS was significantly correlated to K(mean) (P<.01) and the horizontal WTW (P<.01) but not to SE (P = .70). CONCLUSION: There was a close relationship between the various corneal parameters, which is significant in refractive surgery and intraocular lens calculation and design. PMID- 17081864 TI - In vivo architectural analysis of 3.2 mm clear corneal incisions for phacoemulsification using optical coherence tomography. AB - PURPOSE: To analyze in vivo the architecture of clear corneal incisions (CCIs) for phacoemulsification using optical coherence tomography (OCT). SETTING: Anterior Segment Department, Asociacion Para Evitar la Ceguera en Mexico, Hospital Dr Luis Sanchez Bulnes, Mexico. METHODS: A prospective masked study analyzed 20 unsutured CCIs placed superiorly and created in a uniplanar fashion with a 3.2 mm slit-angled metal keratome. All wounds were evaluated with a retinal OCT model 1, 3, and 30 days postoperatively. Intraocular pressure (IOP) and incision leakage were checked. The architecture was described according to the angle of incidence, apposition of the epithelial and endothelial margins, and wound sealing. RESULTS: No leakage was detected. The angle varied from 33 to 85 degrees; angles greater than 75 degrees were done by a surgeon in training. Wound apposition at the epithelial margin was achieved in all cases. In contrast, imperfect apposition of the endothelial margin was seen in 45% of incisions on day 1 and in 15% on day 30. Incomplete sealing of the wound was seen by OCT in 25% of cases at 24 hours and persisted in 10% of all cases at 1 month. This gaping occurred on the endothelial side and never translated to the epithelial margin. No statistical correlation was found between gaping and the angle of the incision, IOP variations, or surgeon experience. CONCLUSIONS: Although in vivo CCIs caused minor anatomic imperfections, they were clinically stable independent of incision angle, IOP variation, and surgeon experience. Incision stability may be related to careful wound construction, epithelial viability, stromal edema, and efficient endothelial pumping. PMID- 17081865 TI - Anterior segment OCT and phakic intraocular lenses: a perspective. AB - Perfect tolerance is expected when one implants a phakic intraocular lens (pIOL) in the anterior segment. Not only should the material be compatible, but the pIOL must respect the anatomy of the anterior chamber. Based on 3 years of experience using an anterior segment optical coherence tomographer (Visante OCT, Carl Zeiss Meditec), I have defined numerous safety criteria for pIOLs. The internal dimensions of the anterior chamber must be considered along different meridians. I propose an objective measurement of the iris dome, the crystalline lens rise (CLR), which is the distance between the anterior pole of the crystalline lens and a line joining the 2 opposite iridocorneal angles. In a series with the Artisan IOL (Ophtec), pigment dispersion syndrome appeared in 70% of cases in which the CLR was greater than 600 microm. Angle-supported IOLs must be placed relative to the anterior chamber's largest diameter; in the same series of cases, the anterior chamber was oval with a large vertical axis in 74% of cases. The posterior face of an angle-supported IOL must have a 700 microm vault to respect the physiological modifications of the crystalline lens. It is difficult to know the posterior chamber's exact diameter as it varies with the horizontal or vertical axis. It also undergoes constant modifications due to accommodation and aging. PMID- 17081866 TI - High-speed optical coherence tomography for management after laser in situ keratomileusis. AB - PURPOSE: To report applications of optical coherence tomography (OCT) in the management of laser in situ keratomileusis (LASIK) related problems. SETTING: Doheny Eye Institute and Department of Ophthalmology, Keck School of Medicine of the University of Southern California, Los Angeles, California, USA. METHODS: Five patients referred for LASIK-related problems were enrolled in a prospective observational study. Clinical examination, ultrasound (US) pachymetry, Placido ring slit-scanning corneal topography (Orbscan II, Bausch & Lomb), and high-speed corneal OCT were performed. RESULTS: In cases of regression and keratectasia, OCT provided thickness measurements of the cornea, flap, and posterior stromal bed. Locations of tissue loss and flap interface planes were identified in a case with a recut enhancement complication. The information was used to determine whether further laser ablation was safe, confirm keratectasia, and manage complications. Optical coherence tomography measurements of central corneal thickness agreed well with US pachymetry measurements (difference 6.4 microm +/- 11.7 [SD]) (P = .026), while Orbscan significantly underestimated corneal thickness (-67.5 +/- 72.5 microm) (P = .17). CONCLUSIONS: High-speed OCT provided noncontact imaging and measurement of LASIK anatomy. It was useful in monitoring LASIK results and evaluating complications. PMID- 17081867 TI - Measuring total corneal power before and after laser in situ keratomileusis with high-speed optical coherence tomography. AB - PURPOSE: To measure total corneal power using optical coherence tomography (OCT). SETTING: Refractive surgery practices at 2 academic eye centers in Cleveland, Ohio, and Los Angeles, California, USA. METHODS: Thirty-two eyes of 17 patients having myopic laser in situ keratomileusis (LASIK) were enrolled in a prospective observational study. Manifest refraction, OCT, and Placido ring corneal topography with the Atlas 995 (Carl Zeiss Meditec, Inc.) were performed preoperatively and 3 months after laser in situ keratomileusis (LASIK). A high speed (2000 axial scans/second) corneal and anterior segment OCT prototype was used. The total corneal power was calculated by summation of the anterior and posterior surface powers, and the value was compared with that determined by simulated keratometry. Two methods of measuring total corneal power were tested: the direct method, which used OCT to measure both corneal surfaces directly, and the hybrid method, which combined OCT with anterior corneal topography. RESULTS: The repeatability (pooled standard deviation) of measuring total corneal power using the hybrid method was 3 times better than that using the direct method. It was 0.23 diopter (D) before LASIK and 0.26 D after LASIK. Preoperative total power was 1.13 D (2.6%) lower than the simulated keratometry. Compared to the LASIK-induced change in spherical equivalent refraction, the change in total corneal power was equivalent, while the change in simulated keratometry power was significantly smaller (-18.8%) (P<.001). CONCLUSIONS: Keratometry using the traditional index of 1.3375 overestimated the total power in preoperative corneas and underestimated LASIK-induced refractive change. Measuring both corneal surfaces using a combination of OCT and Placido ring topography provided a better measure of total corneal power that closely tracked the refractive change in post LASIK eyes. PMID- 17081868 TI - Corneal-thickness spatial profile and corneal-volume distribution: tomographic indices to detect keratoconus. AB - PURPOSE: To evaluate whether the corneal-thickness spatial profile and corneal volume distribution differentiate keratoconic corneas from normal corneas using new tomography parameters. SETTING: Subspecialty cornea and refractive practice, Fluminense Federal University, Rio de Janeiro, Brazil. METHODS: Forty-six eyes diagnosed with mild to moderate keratoconus and 364 normal eyes were studied by the Pentacam Comprehensive Eye Scanner. Corneal thickness at the thinnest point and the averages of the points on 22 imaginary circles centered on the thinnest point with increased diameters at 0.4 mm steps were calculated to create a corneal-thickness spatial profile. Corneal volume was calculated within diameters from 1.0 to 7.0 mm with 0.5 mm steps centered on the thinnest point to create the corneal-volume distribution. The percentage increase in thickness and the percentage increase in volume were calculated for each position of the corneal thickness spatial profile and corneal-volume distribution from their first value. Statistical analysis was done using the Wilcoxon 2-independent-sample test to compare mean levels using S-Plus-4.0 software (MathSoft) and a normal linear model under a Bayesian frame for estimating the mean variation in thickness and volume using the BUGS 0.6 package. RESULTS: Statistically significant differences were observed between the groups (P<.05) in all positions of corneal-thickness spatial profile and corneal-volume distribution and in the percentage increase in thickness and percentage increase in volume between 3.5 mm and 7.0 mm diameters. CONCLUSIONS: Corneal-thickness spatial profile, corneal-volume distribution, percentage increase in thickness, and percentage increase in volume were different between keratoconic corneas and normal corneas and could serve as indices to diagnose keratoconus and screen refractive candidates. Further studies are necessary to evaluate whether these tomographic indices are more sensitive and specific than the classic Placido-based topography. PMID- 17081869 TI - Optical coherence tomography to assess intrastromal corneal ring segment depth in keratoconic eyes. AB - PURPOSE: To investigate intrastromal corneal ring segment depth with a high-speed corneal optical coherence tomography (OCT) system. SETTING: Doheny Eye Institute, University of Southern California, Los Angeles, California, USA. METHODS: A prospective observational case series comprised 4 eyes of 4 patients receiving Intacs intrastromal corneal ring segments (Addition Technology, Inc.) for keratoconus. Optical coherence tomography (OCT) was performed between 7 days and 43 days after implantation. RESULTS: The slitlamp impression of intrastromal corneal ring segment implantation depth did not correlate well with OCT measurements (r(2) = 0.68). The fractional implantation depth was correlated with several surgical variables using a stepwise multivariate regression model, and 2 statistically significant correlations were found. The position of the distal portions of the ring segments was shallower than that of the portion closer to the insertion site (P = .003). Segments placed in the inferior cornea (P = .008) experienced more distal shallowing. Shallower depth was associated with greater fractional anterior stromal compression (P = .04). CONCLUSIONS: Shallower placement of intrastromal corneal ring segments may result in more complications, such as epithelial-stromal breakdown and extrusion, because of the greater anterior stromal tensile strain. The distal and inferior portions of intrastromal corneal ring segments tended to be placed at a shallower depth. Optical coherence tomography provided precise measurement of ring segment depth and may help identify implants that pose a greater risk for depth-related complications. PMID- 17081870 TI - Anterior segment imaging using optical coherence tomography and ultrasound biomicroscopy in secondary pigmentary glaucoma associated with in-the-bag intraocular lens. AB - A 43-year-old man with high myopia developed unilateral pigmentary glaucoma with recurrent episodes of painless blurred vision in the left eye following uneventful phacoemulsification. Bilateral cataract surgery was performed with capsular bag implantation of a +4.00 diopter AcrySof MA60 intraocular lens (IOL) (Alcon Laboratories) followed by a neodymium:YAG laser capsulotomy. Secondary iatrogenic dispersion syndrome in the left eye with subsequent intraocular pressure elevation was suspected in the presence of anterior chamber pigmented cells, circular epithelial iris loss around the pupil, and trabecular hyperpigmentation. Close contact between the edge of the IOL and the posterior pigmented iris epithelium, which was clinically suggested by anterior biomicroscopy, was documented by ultrasound biomicroscopy and optical coherence tomography of the anterior segment. PMID- 17081871 TI - Combined Artemis very high-frequency digital ultrasound-assisted transepithelial phototherapeutic keratectomy and wavefront-guided treatment following multiple corneal refractive procedures. AB - We present a patient with severe visual symptoms following multiple corneal refractive procedures including automated lamellar keratoplasty, arcuate keratotomy, laser in situ keratomileusis (LASIK), LASIK enhancement by cutting another flap, followed by a further LASIK enhancement by flap lifting. Topography was irregularly irregular, and the best spectacle-corrected visual acuity (BSCVA) was 20/25. Artemis very high-frequency digital ultrasound (US) analysis of the anatomical irregularities of the epithelium and stroma was used in conjunction with the topography to determine the cause of the visual symptoms. Very high frequency digital US-assisted transepithelial phototherapeutic keratectomy together with a wavefront-guided treatment was used to reduce the stromal surface irregularities and the higher-order aberrations (HOAs), respectively. The treatment successfully regularized the stromal surface, dramatically reduced the HOAs, improved the contrast sensitivity to the high normal range, and improved the BSCVA to 20/20. PMID- 17081872 TI - Artemis very high-frequency digital ultrasound-guided repositioning of a free cap after laser in situ keratomileusis. AB - We present a patient in whom a symmetrically round free cap occurred during laser in situ keratomileusis (LASIK), and flap repositioning was performed without laser ablation. A loss of 3 lines of best spectacle-corrected visual acuity (BSCVA), monocular diplopia, and topographic irregular astigmatism confirmed that the free cap orientation was incorrect. Two subsequent free cap rotations based on refraction failed to realign the free cap into its original position. Artemis 3-dimensional very high-frequency digital ultrasound analysis found the thickness profiles of the free cap and bed to be irregular and mismatched. The rotation required for anatomic realignment was determined by digitally generating a "lock and key" superimposition of the free cap and stromal bed thickness profiles. After Artemis-guided free cap rotation, the eye regained preoperative BSCVA and symmetrical corneal topography with a +0.50 diopter change in spherical equivalent. PMID- 17081873 TI - Direct residual stromal thickness measurement for assessing suitability for LASIK enhancement by Artemis 3D very high-frequency digital ultrasound arc scanning. AB - We present a patient scheduled for LASIK enhancement based on conventional residual stromal thickness (RST) prediction methods in whom direct measurement of the RST changed the management due to an unexpectedly low RST. The preoperative refraction was -6.00 -0.50 x 115 in the right eye and -6.00 -0.50 x 20 in the left eye. At 9 months, the refractions had regressed to -0.50 -0.50 x 150 and 0.75 -0.25 x 145, respectively. Predicted RST based on preoperative parameters was 283 microm in the right eye and 281 microm in the left eye, sufficient for the planned enhancement. Using the Artemis 3-dimensional very high-frequency digital ultrasound arc scanner, the minimum RST was directly measured as 277 microm in the right eye but only 212 microm in the left eye, which may have significantly increased the risk of iatrogenic ectasia yielding a predicted post enhancement RST of 253 microm and 192 microm, respectively. The treatment plan was altered as a result of the thinner than predicted RST in the left eye; an enhancement was performed in the right eye only. A second Artemis examination after 22 months found the RST in the left eye to be stable. PMID- 17081874 TI - Camera adapter for anterior segment slitlamp photography. AB - We describe an original, affordable and readily accessible slit lamp camera adapter. This is easily assembled from parts already available in most eye units, allowing ready access to anterior segment photography, using both digital and analog cameras. PMID- 17081875 TI - In vivo imaging of posterior capsule opacification using Spectral Optical Coherence Tomography. AB - Three years after uneventful extracapsular cataract extraction with implantation of a poly(methyl methacrylate) (PMMA) intraocular lens (IOL) (MZ60BD, Alcon) in a 74-year-old man, the anterior segment of the right eye was photographed with a Canon EOS 300D digital camera and examined with a slitlamp and a prototype spectral optical coherence tomography (SOCT) instrument. Subsequently, a neodymium:YAG laser posterior capsulotomy was performed, followed by another examination. The quality of the SOCT images was adequate for detailed cross sectional evaluation of the IOL, posterior capsule opacification (PCO), and morphological changes after laser capsulotomy. En face contour maps of PCO distribution were created from 3-dimensional SOCT data. The results presented indicate future applicability of SOCT technology in evaluating modern IOL designs and investigating the process of PCO formation. PMID- 17081876 TI - Thermal comparison of the Bausch & Lomb Millennium, Alcon AdvanTec Legacy, and the AMO Sovereign WhiteStar phacoemulsification systems. PMID- 17081877 TI - Effect of change in central lens thickness and lens shape on age-related decline in accommodation. PMID- 17081879 TI - Coma, trefoil, and double-angled plots. PMID- 17081882 TI - Managing the IOL-CTR-CB complex. PMID- 17081883 TI - Intraoperative floppy-iris syndrome associated with tamsulosin. PMID- 17081884 TI - Anterior chamber collapsing technique for soft lenses. PMID- 17081885 TI - Fugo blade to enlarge phimotic capsulorhexis. PMID- 17081886 TI - Use of triamcinolone acetonide in retained lens surgery. PMID- 17081887 TI - Inadvertent insertion of an opposite-power intraocular lens. PMID- 17081888 TI - Consultation section. Refractive surgical problem. PMID- 17081894 TI - Scleral fixation without conjunctival dissection. AB - Scleral fixation of intraocular lenses (IOLs) and adjunctive capsular devices can be performed under the protection of a scleral flap. A modification of this technique uses a scleral pocket initiated through a peripheral clear corneal incision. Full-thickness passage of a double-armed suture through the scleral pocket and conjunctiva, with subsequent retrieval of the suture ends through the external incision for tying, facilitates scleral fixation. This modification offers several advantages over traditional methods: It eliminates the need for conjunctival dissection and scleral cauterization; a scleral pocket affords a greater surface area for suture placement through an ab externo or ab interno approach; retrieval of the sutures through the external corneal incision and subsequent tying allows the suture knot to pass under the protective roof of the scleral pocket, negating the need for suture knot rotation; and the architecture of the scleral pocket eliminates the need for sutured wound closure. Suture retrieval and scleral fixation through a corneoscleral pocket offers a refined method for fixation of IOLs and other intraocular adjunctive devices. PMID- 17081895 TI - Surgical technique for congenital iris coloboma repair. AB - We describe a surgical technique for managing congenital iris coloboma. After phacoemulsification with placement of an intraocular lens in the capsular bag, coloboma repair is begun by bisecting the iris sphincter on both sides of its attachment near the chamber angle. The iris leaflets central to the sphincterectomies are approximated using a 10-0 polypropylene suture (Prolene, Ethicon, Inc.) and a modified Siepser pupilloplasty technique. The remaining peripheral iris defect is closed in a similar fashion. In patients with congenital iris coloboma, phacoemulsification with in-the-bag IOL implantation followed by this pupilloplasty technique was effective in providing functional and cosmetic repair of a congenital iris coloboma. PMID- 17081896 TI - Microincision bimanual phacotrabeculectomy in eyes with coexisting glaucoma and cataract. AB - PURPOSE: To report the technique and surgical outcomes of microincision bimanual phacotrabeculectomy in eyes with glaucoma and coexisting cataract. SETTING: Glaucoma Service, Hong Kong Eye Hospital, Hong Kong SAR, China METHODS: Microincision bimanual phacoemulsification with a sleeveless phaco needle and irrigating chopper was performed in combination with trabeculectomy. Phacoemulsification was performed through 2 small clear corneal wounds, sparing the trabeculectomy site from trauma. The intraocular lens (IOL) was then implanted through the trabeculectomy site, so no corneal wound larger than 1.5 mm was required for IOL implantation. RESULTS: The first 10 consecutive eyes of 10 patients who had combined phacotrabeculectomy by a microincision bimanual technique had significantly reduced intraocular pressure. There was a 10.5% reduction in the mean corneal endothelial cell count. There were no other complications up to 6 months after surgery. CONCLUSION: Microincision bimanual phacotrabeculectomy appeared to be an effective and safe option in patients with glaucoma and coexisting cataract. PMID- 17081898 TI - Comparison of 2 multiple-measurement infrared pupillometers to determine scotopic pupil diameter. AB - PURPOSE: To compare a monocular and a binocular multiple-measurement digital infrared pupillometers for measuring scotopic pupil diameter. SETTING: Department of Ophthalmology, Loma Linda University, Loma Linda, California, USA. METHODS: Scotopic pupil size was measured after 1 minute of dark adaptation in 42 eyes of 21 volunteers. Measurements were taken twice each with 2 multiple-measurement digital infrared pupillometers, the monocular pupillometer (Neuroptics, Inc.), and the binocular pupillometer (P2000D, Procyon, Ltd.) Intraclass correlation coefficients (ICC) and limits of agreement (LOA) were used to measure repeatability and agreement of measures with each instrument and between instruments. The Wilcoxon signed rank test was used to compare variability of measurements within each instrument. RESULTS: The mean scotopic pupil size was 4.79 mm +/- 0.95 (SD) with the Procyon and 4.86 +/- 0.93 mm with the Neuroptics. Repeatability and agreement tests for the Procyon measures showed the following: ICC, 0.954; 95% confidence interval (CI), 0.916-0.975; LOA, -0.60 to 0.56; range, 1.16. The Wilcoxon signed rank test of variability gave a Z score of -2.53 (P = .01, 2 tailed). The repeated measures testing with the Neuroptics pupillometer showed the following: ICC, 0.985; 95% CI, 0.972-0.992; LOA, -0.39 to 0.26; range, 0.64; Z score, -1.15 (P = .25, 2-tailed). Repeatability and agreement tests for measures between instruments showed the following: ICC, 0.954; 95% CI, 0.916 0.975; LOA, -0.60 to 0.50; range 1.11. CONCLUSIONS: There was a high repeatability and agreement in scotopic pupil diameter for repeated measures within each device and measurements between the devices. Differences in variability in scotopic pupil diameter evaluated by the Wilcoxon signed rank test were significant only with the Procyon pupillometer. PMID- 17081897 TI - Inhibition of intraocular fibrin formation after infusion of low-molecular-weight heparin during combined phacoemulsification-trabeculectomy surgery. AB - PURPOSE: To evaluate the effect of intraocular infusion of dalteparin, a low molecular-weight heparin (LMWH) administered in the infusion fluid to prevent early postoperative fibrin formation in combined phacoemulsification trabeculectomy surgery (phacotrabeculectomy). SETTING: Farabi Eye Hospital, Tehran, Iran. METHODS: Standard combined phacotrabeculectomy was performed prospectively in a masked fashion in 60 eyes; 35 eyes received 5 IU/mL LMWH in the infusate and 25 eyes served as controls. Masked postoperative examinations assessed visual acuity, intraocular pressure (IOP), anterior chamber cell and flare, and fibrin formation with slitlamp examinations 1, 3, 7, 30, and 90 days after surgery. RESULTS: The rate of intraocular fibrin/membrane formation was significantly lower in the LMWH group than in the control group at each follow-up (P<.001, Student t test) . The amount of IOP was lower in LMWH group at each follow up (P<.01, Student t test). Final visual acuity was better in LMWH group (P = .03, Student t test). The frequency of synechia formation was lower in the LMWH group on day 90 (P = .002, chi-square test). CONCLUSIONS: Infusion of dalteparin, a LMWH, may be an effective inhibitor of postoperative fibrin formation in phacotrabeculectomy surgery. It may reduce the frequency of synechia formation and improve visual acuity. It may not be associated with increased risk for intraoperative or postoperative complications at the tested dose. This drug may be a useful adjunct in combined surgery. PMID- 17081899 TI - Postoperative surface deposits on intraocular lenses in children. AB - PURPOSE: To report the clinicopathologic features of 4 intraocular lenses (IOLs) composed of 3 different hydrophilic biomaterials explanted from children who had postoperative opacification of the IOL optic. SETTING: David J Apple, MD, Laboratories for Ophthalmic Devices Research, John A. Moran Eye Center, Department of Ophthalmology and Visual Sciences, University of Utah, Salt Lake City, Utah, USA. METHODS: The IOLs were explanted 20, 11, 22, and 25 months postoperatively from children aged 10, 3, 36, and 20 months old, respectively, at IOL implantation. Clinical data were obtained to correlate the findings with possible associated risk factors. The explanted IOLs were examined by gross and light microscopy. They were further analyzed with a stain for calcium, alizarin red 1%. Scanning electron microscopy and energy dispersive X-ray spectroscopy (EDS) were also performed. RESULTS: The primary reason for cataract surgery in Case 1 and Case 2 was persistent hyperplastic primary vitreous (PHPV); 1 patient received a B-Lens IOL (Hanita) and the other a Centerflex IOL (Rayner). The primary reason in Case 3 was familial bilateral congenital cataract and in Case 4, rubella cataract; both patients received a Hydroview IOL (Bausch & Lomb). All 4 IOLs had surface deposits on the optic, but the morphology of the deposits on the B-Lens and Centerflex IOLs was different than that in previously reported cases. The deposits in all four cases stained positive with alizarin red and consisted of calcium and phosphorus when imaged with EDS. CONCLUSIONS: Calcified deposits on 2 Hydroview IOLs explanted from children were similar to those seen in adults with the same IOL. The deposits on the B-Lens and Centerflex IOLs were probably secondary to a breakdown of the blood-aqueous barrier caused by preexisting PHPV. PMID- 17081900 TI - Secondary diffractive bifocal piggyback intraocular lens implantation. AB - PURPOSE: To assess the efficacy and safety of implanting a bifocal diffractive intraocular lens (IOL) using the piggyback technique to provide pseudoaccommodation. SETTING: Instituto Oftalmologico Fernandez-Vega, Oviedo, Spain. METHODS: This prospective noncomparative case series included 6 pseudophakic emmetropic patients who had a monofocal IOL implanted in the capsular bag. All eyes had secondary piggyback Acri. Twin bifocal diffractive IOL (Acri.Tec) implantation in the ciliary sulcus to provide pseudoaccommodation. Contralateral implantation of 1 Acri. Twin near-weighted 733D IOL in the nondominant eye and 1 Acri. Twin distance-weighted 737D IOL in the dominant eye was performed. Monocular and binocular best distance-corrected visual acuity and distance-corrected near visual acuity were evaluated 6 months after surgery. RESULTS: The mean binocular best distance-corrected visual acuity and distance corrected near visual acuity were -0.080 +/- 0.056 logMAR and -0.016 +/- 0.037 logMAR, respectively. There were no statistically significant differences in binocular best distance-corrected acuity after the piggyback IOL implantation (P>.01). The differences between monocular and binocular visual acuity were the result of the distance- and near-weighted light distribution of the Acri. Twin IOLs. All IOLs were well centered with no tilt. Two eyes had pupillary capture of the optic. CONCLUSIONS: The Acri. Twin bifocal diffractive IOL implanted in the ciliary sulcus using the piggyback technique provided pseudoaccommodation in emmetropic pseudophakic eyes. Binocular implantation of these IOLs should be performed considering the differences in light distribution of the distance weighted IOL and the near-weighted IOL models. PMID- 17081901 TI - Longitudinal changes in subjective and objective visual function 5 years after cataract surgery Prospective population-based study. AB - PURPOSE: To investigate the long-term outcomes of cataract surgery by analyzing data collected 5 years after surgery and comparing with preoperative and postoperative subjective and objective visual function results. SETTING: Norrlands University Hospital, Umea, Sweden. METHODS: A prospective longitudinal population-based cohort study comprised 810 patients who had cataract surgery during a 1-year period within a geographically defined area. Evaluated were visual acuity data and Visual Function-14 questionnaire (VF-14) results before and after surgery. Five years later, the 590 patients still alive were offered eye examinations and asked to fill out the questionnaire. RESULTS: Of the 590 patients asked to participate at 5 years, 530 answered the questionnaire and 467 had eye examinations. The median VF-14 total score for all patients after surgery was 100; at 5 years, the score decreased to 96.7 (P = .001). Five years after surgery, 46% of patients had unchanged or better visual acuity in the operated eye, 37% had lost more than 0.1 logMAR unit, and 22% had a reduction in VF-14 score of 10 points or more. The two main reasons for the decline in visual acuity and VF-14 scores were age-related macular degeneration (ARMD) (47% and 60%, respectively) and glaucoma (12% and 11%, respectively). Age, co-morbidity, and VF 14 scores after surgery were independently associated with the VF-14 score 5 years after surgery. CONCLUSIONS: Subjective and objective visual function 5 years after cataract surgery remained stable in most patients. Co-morbidity, most commonly ARMD, was the most frequent cause of deterioration of visual acuity and decrease in VF-14 scores. Age and co-morbidity were independently associated with the VF-14 score 5 years after surgery. PMID- 17081902 TI - Sedative effect of acupuncture during cataract surgery: prospective randomized double-blind study. AB - PURPOSE: To assess the effectiveness of acupuncture in reducing anxiety in patients having cataract surgery under topical anesthesia. SETTING: Vita-Salute University of Milan and IRCCS H. San Raffaele, Milan, Italy. METHODS: In a prospective randomized double-blind controlled trial, anxiety levels before and after cataract surgery in 3 groups (A = no acupuncture, B = true acupuncture starting 20 minutes before surgery, C = sham acupuncture starting 20 minutes before surgery) were compared using the Visual Analog Scale (VAS). Twenty-five patients scheduled for inpatient phacoemulsification were enrolled in each group. All surgeries were performed using topical anesthesia. Exclusion criteria were refusal to provide informed consent, use of drugs with sedative properties, psychiatric disease, pregnancy, knowledge of the principles of acupuncture, anatomic alterations, or cutaneous infections precluding acupuncture at the selected acupoints. RESULTS: Preoperative anxiety levels were significantly lower only in Group B (P = .001). Anxiety in Group B was significantly lower than in Group A (P = .001) and Group C (P = .037). Regarding postoperative anxiety, the mean VAS score was 39 +/- 5 in Group A, 19 +/- 3 in Group B, and 31 +/- 4 in Group C. The difference was significant only between Group A and Group B (P = .003). CONCLUSION: Acupuncture was effective in reducing anxiety related to cataract surgery under topical anesthesia. PMID- 17081903 TI - After-cataract evaluation after using balanced salt solution, distilled deionized water, and 5-fluorouracil with a sealed-capsule irrigation device in the eyes of 4-week-old rabbits. AB - PURPOSE: To evaluate the Perfect Capsule sealed-capsule irrigation device (Milvella Pty., Ltd.) using 3 substances in young rabbit eyes. SETTING: St. Erik's Eye Hospital, Stockholm, Sweden. METHODS: Thirty 4-week-old rabbits had clear lens extraction in both eyes. In 1 randomly selected eye, the Perfect Capsule was applied and the lens capsule was irrigated for 5 minutes with 1 of 3 substances: balanced salt solution (BSS), distilled deionized water (DDW), or 5 fluorouracil (5-FU) 50 mg/mL. In the other eye, no sealed capsule irrigation was used. Forty days postoperatively, the animals were killed and the eyes fixed in formalin for histologic analysis. After-cataract was evaluated in 3 ways: clinically, from photographs, and histologically. Central posterior capsule thickness was evaluated using a microscope, camera, and computer. RESULTS: The Perfect Capsule sealed-capsule irrigation system could be used in all selected eyes. The vacuum to the anterior capsule was tight, and the system was sealed in all eyes. After-cataract developed in the BSS group and DDW group, but not in the 5-FU group. The 5-FU group had significantly less after-cataract than the other 2 groups (P<.05). There was no difference between the groups in capsule thickness. CONCLUSIONS: The Perfect Capsule sealed-capsule irrigation system could be used in small eyes. Distilled deionized water did not prevent after-cataract in rabbit eyes with highly proliferative cells, but 5-FU was effective in preventing after cataract. PMID- 17081904 TI - AcrySof Natural intraocular lens optical characteristics during and after different doses of ultraviolet-visible light illumination. AB - PURPOSE: To determine the optical transmission properties of a recently introduced intraocular lens (IOL) (AcrySof Natural SN60AT, Alcon Laboratories) and compare them with those of a conventional IOL (AcrySof SA60AT, Alcon Laboratories) after ultraviolet (UV)-visible light irradiation. SETTING: Eye Clinic and Department of Clinical Physiopathology, University of Florence, and the National Institute of Applied Optics, Florence, Italy. METHODS: Two IOLs, the AcrySof Natural with a UV and blue-light filter and the AcrySof without a blue light filter, were studied. Three IOLs of each model with different dioptric powers were examined in two experimental setups. The samples were stressed with very high irradiance to reproduce light aging, and the transmission properties of the IOLs were studied. RESULTS: No significant decrease in transmission was found at a UV-visible light dose equivalent to 100 years of light aging. CONCLUSION: The AcrySof Natural IOL has stable optical characteristics. PMID- 17081905 TI - Two-step laser in situ keratomileusis to correct high astigmatism after corneal autograft. AB - We report a case in which significant anisometropia and astigmatism after an autograft by rotation was treated by a 2-step laser in situ keratomileusis (LASIK) procedure. Six weeks after the lamellar keratotomy, photoablation was performed using the LADARVision 4000 excimer laser. The ablation depth was 87.2 mum on a 6.0 mm optical zone. One day postoperatively, the uncorrected visual acuity (UCVA) was 7/10 and the refractive error was -0.25 -0.25 x 67. Two months later, the UCVA was 5/10 and the best corrected visual acuity, 8/10 Parinaud 2 with a refractive error of +1.75 -1.25 x 16. The case demonstrates the effectiveness of 2-step LASIK for the correction of astigmatism induced by a corneal autograft. The significant reduction in autograft-induced astigmatism, together with stabilization of the higher-order aberrations, resulted in restoration of satisfactory functional vision. PMID- 17081906 TI - Secondary prosthetic iris implantation following traumatic total aniridia and pseudophakia. AB - We present 2 pseudophakic patients who had traumatic episodes that resulted in total expulsion of the iris without disturbing the intraocular lens (IOL). Because of intolerable glare, each patient was managed by reopening the fibrosed capsular bag and implanting 2 multi-finned prosthetic iris devices through a small incision, leaving the IOL in place. Following surgery, glare was no longer present and excellent visual acuity was maintained. We believe these are the first pseudophakic patients with traumatic total aniridia to be managed by this approach. PMID- 17081907 TI - Shifting crystalline pseudohypopyon secondary to lens absorption with spontaneous openings in the anterior lens capsule. AB - This is the first reported case of a crystalline pseudohypopyon presenting as a layer of "snowdrift deposits" that settled inferiorly in the anterior chamber of the eye of a 55-year-old man and was associated with freely floating crystals in the aqueous humor, producing a "snowy Christmas Eve" appearance. The cause was spontaneous openings in the center of the anterior capsule with leaching of the crystalline substance into the anterior chamber. A dense membranous cataract was left after the lens material was partly absorbed. There was no history of trauma, surgery, or known systemic disease, and the patient presented with a unilateral, mobile, 4.0 mm white pseudohypopyon with no global pain, photophobia, or lacrimation. Although the eye was not injected on admission, repetitive head shaking resulted in dusky-red perilimbal hyperemia. A B-mode ultrasound revealed an advanced retinal detachment, and electroretinographic recordings were undetectable. Visual acuity decreased to hand motions in the affected eye, and the patient ultimately required anterior chamber paracentesis. Such sediments should be differentiated from other forms of true or pseudohypopyons because the course, treatment, and prognosis are different for each. PMID- 17081908 TI - Capsular block syndrome presenting with a hyperopic shift. AB - We report a case of late postoperative capsular block syndrome presenting with a hyperopic shift and discuss possible causes of this unusual presentation. PMID- 17081909 TI - Cancer care: WHO's poor relation. PMID- 17081910 TI - Keynote comment: cancer survivorship and ageing--a double whammy. PMID- 17081911 TI - Rising importance of patient-reported outcomes. PMID- 17081912 TI - Clarifying breast cancer risks associated with menopausal hormone therapy. PMID- 17081913 TI - Multidisciplinary team meetings: where is the value? PMID- 17081914 TI - Care with intrathecal trastuzumab. PMID- 17081915 TI - Patient versus clinician symptom reporting using the National Cancer Institute Common Terminology Criteria for Adverse Events: results of a questionnaire-based study. AB - BACKGROUND: The Common Terminology Criteria for Adverse Events (CTCAE) are used as standard practice in trials of cancer treatments by clinicians to elicit and report toxic effects. Alternatively, patients could report this information directly as patient-reported outcomes, but the accuracy of these reports compared with clinician reports remains unclear. We aimed to compare the reporting of symptom severity reported by patients and clinicians. METHODS: Between March and May, 2005, a questionnaire with 11 common CTCAE symptoms was given to consecutive outpatients and their clinicians (physicians and nurses) in lung and genitourinary cancer clinics in the Memorial Sloan-Kettering Cancer Center, New York, NY, USA. Patients completed a version that used language adapted from the CTCAE for patient self-reporting. The results from the questionnaire were compared with clinician reporting of the same symptoms. FINDINGS: Of 435 patients and their clinicians asked to take part in the study, 400 paired surveys were completed. For most symptoms, agreement between patient and clinician was high, and most discrepancies were within a grade difference of one point. Agreement was higher for symptoms that could be observable directly, such as vomiting and diarrhoea, than for more subjective symptoms, such as fatigue and dyspnoea. Differences in symptom reporting rarely would have changed treatment decisions or dosing, and patients assigned greater severity to symptoms more than did clinicians. No significant differences were recorded between the results when the questionnaire was completed by the patient before or after the clinician. INTERPRETATION: Patient reporting of symptoms could add to the current approach to symptom monitoring in cancer treatment trials. Future research should assess the effect of self reporting on clinical outcomes and efficiency, and the use of real-time collection of patient-reported outcomes for early detection of potentially serious adverse events. PMID- 17081916 TI - Hormonal therapy for menopause and breast-cancer risk by histological type: a cohort study and meta-analysis. AB - BACKGROUND: Little information is available on how the risk of breast cancer associated with the use of hormone therapy for menopause varies by histological type. We aimed to describe such associations for eight histological types of breast cancer. METHODS: Analyses are based on 1 031 224 postmenopausal women recruited in 1996-2001 into a nationwide UK cohort study, and followed for incident cancer and death. Relative risks associated with use of hormone therapy were estimated for eight histological types of breast cancer. FINDINGS: During 3.6 million person-years of follow-up, 14 102 breast cancers were diagnosed, of which 13 782 (98%) had histological type recorded: 11 869 (86%) were invasive, including 8007 ductal, 1526 lobular, 365 mixed ductal-lobular, 492 tubular, 71 medullary, and 148 mucinous cancers; and 1913 (14%) were in situ, including 1443 ductal and 86 lobular cancers. The relative risks of invasive breast cancer in current users compared with never users of hormone therapy varied significantly according to tumour histology overall (p<0.0001), for users of oestrogen-only therapy (p=0.0001), and for users of oestrogen-progestagen therapy (p<0.0001). The largest relative risks in current compared with never users of hormone therapy were seen for lobular (relative risk 2.25, 95% CI 2.00-2.52), mixed ductal-lobular (2.13, 1.68-2.70), and tubular cancers (2.66, 2.16-3.28). The relative risks for ductal and mucinous cancers were 1.63 (95% CI 1.55-1.72) and 1.58 (1.08-2.31), respectively. The risk of medullary cancer was not increased (0.74, 0.43-1.28). The relative risk of in-situ disease in current users compared with never users of hormone therapy also varied significantly according to histological type (p=0.03), with a relative risk for lobular carcinoma in situ of 2.82 (1.72-4.63) and 1.56 (1.38-1.75) for ductal carcinoma in situ. The effects of hormone therapy on invasive ductal, lobular, and tubular cancer were generally greater for oestrogen-progestagen therapy than for oestrogen-only therapy, and were attenuated with increasing body-mass index (BMI). INTERPRETATION: The risks associated with use of hormone therapy for menopause differ by histological type of breast cancer, and are substantially attenuated with increasing BMI. PMID- 17081917 TI - Complete metastasectomy in patients with stage IV metastatic melanoma. AB - Patients with stage IV melanoma have traditionally been managed with various systemic treatments; however, overall survival with this approach has been disappointing. Findings of many retrospective, single-institution, and multicentre studies suggest that participants treated with complete metastasectomy for stage IV metastases have enhanced overall 5-year survival. Complete surgical resection of metastatic disease to stage IV sites-including skin, soft tissue, distant lymph nodes, lungs, or other non-CNS visceral regions offers the best chance for prolonged survival. This Review will present data lending support to the idea that if complete surgical metastasectomy is technically feasible, then surgery should be the first option for properly selected patients with stage IV melanoma. PMID- 17081918 TI - Mechanisms of transcoelomic metastasis in ovarian cancer. AB - Metastasis from epithelial ovarian cancer can occur via the transcoelomic, haematogeneous, or lymphatic route. Of these, transcoelomic metastasis is the most common, and is responsible for the greatest morbidity and mortality in women with this disease. Unfortunately, very little is known about the mechanisms behind this process. This review assesses the current evidence and ideas about the biology of transcoelomic dissemination. The mechanisms of cell detachment, migration, and implantation in transcoelomic metastasis are placed within the context of clinical observations of ovarian cancer to derive a stepwise hypothesis of this process. Evidence for transcoelomic dissemination versus transcoelomic metaplasia in ovarian cancer is presented. Future high throughput microarray studies that compare changes at a genomic and gene expression level between primary ovarian tumours and their peritoneal metastases are hoped to lead to a more conclusive picture of transcoelomic metastasis, and to delineate the key molecular players in this process. These studies might also result in the identification of potential new therapeutic targets in ovarian cancer. PMID- 17081919 TI - Multidisciplinary teams in cancer care: are they effective in the UK? AB - Cancer care can be complex, and given the wide range and numbers of health-care professionals involved, an enormous potential for poor coordination and miscommunication exists. Multidisciplinary teams (MDTs) should improve coordination, communication, and decision making between health-care team members and patients, and hopefully produce more positive outcomes. This review describes the many practical barriers to the successful implementation of MDT working, and shows that despite an increase in the delivery of cancer services via this method, research showing the effectiveness of MDT working is scarce. PMID- 17081920 TI - Telling the truth to patients with cancer: what is the truth? AB - Attitudes and practices of truth-telling to people with cancer have shifted substantially in the past few years. However, cultural and individual differences persist, and some difficulties common to all medical specialties are magnified in oncology. In this Personal View, I review and analyse data for attitudes and practices of truth-telling worldwide. I also assess ethical justifications, with special reference to interpersonal aspects of patients' autonomy and the dynamic nature of truth in the clinical context. Examples are provided to show how this ethical perspective can help oncologists to frame the discourse on truth-telling and to find solutions to the dilemmas of whether, when, and how to tell the truth to their patients in clinical practice. Finally, I identify future targets for research. PMID- 17081921 TI - Multidisciplinary care in oncology: medicolegal implications of group decisions. AB - Consensus is growing that multidisciplinary meetings (MDMs) provide the best means of formulating comprehensive treatment plans for patients with cancer. Although many doctors attend MDMs and contribute to the decision-making process, only a few will become involved in a patient's care after the team meeting. Despite this, if a patient was grieved by a decision made in a MDM and wished to recover damages, all doctors present at the meeting would be personally accountable for decisions related to their area of expertise. Doctors should be made aware of the legal implications of their participation in such meetings. A greater awareness of these responsibilities and improved team dynamics should optimise outcomes for patients while limiting exposure of the participants to legal liability. Special attention should be given to providing patients with adequate information in this combined speciality setting. PMID- 17081922 TI - Use of sirolimus for Epstein-Barr virus-positive smooth-muscle tumour. PMID- 17081923 TI - Clinical response of cutaneous squamous-cell carcinoma to bortezomib given for myeloma. PMID- 17081924 TI - Neuron-specific enolase expression in multiple myeloma. PMID- 17081925 TI - Help-seeking behaviors of women with urinary incontinence: an integrative literature review. AB - Urinary incontinence (UI) affects approximately 17 million American women. Women with UI have reported a lower quality of life than women who do not experience UI. In many cases, UI affects a woman's ability to take part in a variety of recreational and social activities, and has negative psychological effects including depression, anxiety, and frustration. Studies show that fewer than 50% of women with UI discuss their symptoms with their health care provider. Although researchers have examined help-seeking behaviors in people with UI, few have examined women of all ages with all types of UI. The purpose of this integrative literature review is to summarize the help-seeking behaviors for UI symptoms among noninstitutionalized women of all ages with all types of UI. Five studies were reviewed. In all of the studies, less than 38% of women sought help for their UI symptoms. Factors affecting help seeking included quality of life scores, perceptions women had regarding the normalcy of UI, and beliefs about treatment options available for the problem. Age, type of UI, severity of UI, and embarrassment were found to be significantly related to help-seeking in most, but not all of the studies. PMID- 17081926 TI - Body image after bilateral prophylactic mastectomy: an integrative literature review. AB - Bilateral prophylactic mastectomy (BPM) can reduce a woman's risk for breast cancer by 80%. Thus, women who are at high risk for familial breast cancer are increasingly opting for BPM as a preventative option. Research indicates that there are psychological benefits to BPM, including a reduction in anxiety about developing breast cancer. The purpose of this integrative review is to summarize the research that has examined the effect of prophylactic mastectomy on women's subsequent body image. Thirteen studies were reviewed. The majority of women were satisfied with their decision. However, the majority of studies indicate that up to one-half of the women suffer a negative effect on body image and changes in sexuality. Knowledge of these findings can improve the practitioner's ability to counsel women regarding this radical decision. Further research, particularly prospective studies, are needed to examine women's body image prior to BPM so that the impact of prophylactic mastectomy can be examined more thoroughly. PMID- 17081927 TI - Reproductive health in the 21st century: two steps forward? PMID- 17081928 TI - Reproductive rights: a call to action. PMID- 17081929 TI - The use of herbs and dietary supplements in gynecology: an evidence-based review. AB - Consumers frequently use herbs and dietary supplements to treat chronic conditions that are poorly responsive to prescription drugs or when prescription drugs carry a high side effect burden. Women may use herbs and supplements for chronic gynecologic conditions, such as menopause, premenstrual syndrome, dysmenorrhea, cyclic mastalgia, and infertility. This review is an evidence-based evaluation of herbs and supplements for these conditions. Therapies that carry a higher level of support from randomized controlled trial evidence include black cohosh for menopause; vitamins B(1) and E for dysmenorrhea; calcium, vitamin B(6), and chasteberry for premenstrual syndrome; and chasteberry for cyclic mastalgia. There were too few trials involving herbs and supplements in infertility to warrant a solid recommendation, but chasteberry, antioxidants, and Fertility Blend have some preliminary support. Midwives may want to consider these alternatives in addition to more traditional treatment options when meeting with patients. PMID- 17081930 TI - Gynecologic and contraceptive services provided by certified nurse-midwives in North Carolina. AB - Gynecologic care by midwives has been little represented in the peer-reviewed literature despite the fact that the majority of midwives provide these services. Offering patients a variety of contraceptive options is important for informed choice and consent and for reducing the unintended pregnancy rate, which is nearly 50% in the United States. This study describes the volume of gynecologic care and the contraceptive methods provided by certified nurse-midwives (CNMs) in North Carolina. The study was conducted using a cross-sectional, confidential, self-administered mail questionnaire completed by 133 CNMs in clinical practice in North Carolina. The majority of the CNMs worked in private practice settings (67%), in urban areas (60%), and had 10 or fewer years of experience (62%). The median number of women seen weekly for gynecologic care was 15 (range 0-80), and 30% of CNMs provided gynecologic care to more than 25 women each week. The contraceptive methods discussed and provided by the CNMs were comprehensive. The high percentage of midwives providing gynecologic care merits further study of the content and quality of this care. PMID- 17081931 TI - Polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is a common endocrine disorder, affecting between 4% and 8% of reproductive aged women. This syndrome, a complex disorder with multiple components, including reproductive, metabolic, and cardiovascular manifestations, has long-term health concerns that cross the life span. The diagnostic criteria for PCOS are ovarian dysfunction evidenced by oligomenorrhea or amenorrhea and clinical evidence of androgen excess (e.g., hirsutism and acne) in the absence of other conditions that can cause these same symptoms. This article reviews current knowledge about the pathophysiology, clinical manifestations, diagnosis, and management of this disorder. PMID- 17081932 TI - Etiology, diagnosis, and management of vaginitis. AB - Women often seek medical care for vaginal complaints. Many times, the cause of the complaint is misdiagnosed by the woman and/or her provider. These vaginal complaints may be related to infections, which when misdiagnosed or mistreated, can lead to more severe problems. This article describes the three most common vaginal infections, bacterial vaginosis, trichomoniasis, and vulvovaginal candidiasis. Appropriate diagnostic techniques and management therapies are reviewed. PMID- 17081933 TI - Ectopic pregnancy. AB - The incidence of ectopic pregnancy in the United States has been reported to be as high as 20 per 1000 pregnancies, a more than 4-fold increase over the last 20 years. Clinical presentation can range from subtle, nonspecific abdominal complaints to acute onset pain or hemorrhagic bleeding. This article reviews the associated risk factors, pathophysiology, diagnosis, and management of ectopic pregnancy. Clinicians must maintain a high index of suspicion whenever women who might be pregnant have abdominal complaints. PMID- 17081934 TI - Ultrasound assessment of the endometrium for irregular vaginal bleeding. AB - Irregular vaginal bleeding is a common symptom of women seeking gynecologic care. Etiologies of irregular vaginal bleeding can be classified into the following categories: pregnancy related (retained products of conception, threatened or missed abortion, or ectopic pregnancy), hormonal (disorders of ovulation, menopause, or hormonal contraceptive use), structural (polyps, myomas, or arteriovenous malformation), neoplasm (endometrial cancer), and infection (endometritis). After the history and physical examination, the initial evaluation of irregular vaginal bleeding has traditionally involved an endometrial biopsy. Transvaginal ultrasound has revolutionized the evaluation of the gynecologic ultrasound examination by providing a minimally invasive means to determine the etiology for the bleeding. Transvaginal ultrasound assessment of the endometrial cavity allows treatment to be tailored to the specific cause of irregular vaginal bleeding, thus saving women time, money, and exposure to unnecessary interventions. The purpose of this article is to give the clinician critical information regarding the capabilities of ultrasound to evaluate women with irregular vaginal bleeding. PMID- 17081935 TI - Special issues in contraception: caring for women with disabilities. AB - Women with disabilities make up a diverse population of underserved clients needing skilled, sensitive, and culturally-competent contraceptive care. Barriers to care for these women are structural, attitudinal, and informational, and lack of attention to their needs is also reflected in limited research and clinical guidelines. Special techniques for a modified gynecologic examination can be employed, and disabled women can work in partnership with providers to promote comfort, respect, and safety. There are specific considerations for the use of different contraceptive methods in women with movement limitations, sensory impairments, seizure disorders, developmental disability, and emotional and psychiatric disorders. A climate of welcoming respect as well as clinical expertise in issues specific to this population can enhance the ability to provide contraception to these women. PMID- 17081936 TI - Current issues in emergency contraception: an overview for providers. AB - Emergency contraception has the potential to greatly reduce the number of unintended pregnancies occurring each year in the United States. Emergency contraception is a safe and effective intervention to which all women should have easy access in the event of an act of unprotected intercourse. Methods of emergency contraception include combined hormone oral contraceptive pills, progestin-only oral contraceptive pills, a dedicated progestin-only emergency contraceptive product, and insertion of a copper intrauterine device. Barriers exist to the increased use of emergency contraception, including the prescription only status of all of the methods and lack of accurate knowledge on the part of health care providers and consumers. This article provides an overview of the clinical management of emergency contraception. PMID- 17081937 TI - The intrauterine device: rethinking old paradigms. AB - The United States continues to have one of the highest rates of unintended pregnancy and elective abortion in developed countries. Intrauterine devices (IUDs) available today offer women safe and highly effective contraception along with noncontraceptive benefits, yet IUDs remain underutilized in part because of outdated and biased information about the risks associated with this method of fertility control. New research demonstrates that IUD use does not increase the risk of pelvic infections or subsequent infertility. IUD use decreases the absolute risk of ectopic pregnancies. In light of this data, the IUD should be made available to women at low-risk for sexually-transmitted infections and should not be denied to women on the basis of parity or marital status. PMID- 17081938 TI - New approaches to fertility awareness-based methods: incorporating the Standard Days and TwoDay Methods into practice. AB - Helping clients select and use appropriate family planning methods is a basic component of midwifery care. Many women prefer nonhormonal, nondevice methods, and may be interested in methods that involve understanding their natural fertility. Two new fertility awareness-based methods, the Standard Days Method and the TwoDay Method, meet the need for effective, easy-to-provide, easy-to-use approaches. The Standard Days Method is appropriate for women with most menstrual cycles between 26 and 32 days long. Women using this method are taught to avoid unprotected intercourse on potentially fertile days 8 through 19 of their cycles to prevent pregnancy. They use CycleBeads, a color-coded string of beads representing the menstrual cycle, to monitor their cycle days and cycle lengths. The Standard Days Method is more than 95% effective with correct use. The TwoDay Method is based on the presence or absence of cervical secretions to identify fertile days. To use this method, women are taught to note everyday whether they have secretions. If they had secretions on the current day or the previous day, they consider themselves fertile. The TwoDay Method is 96% effective with correct use. Both methods fit well into midwifery practice. PMID- 17081939 TI - Women's barrier contraceptive methods: poised for change. AB - Over the past 20 years, the number of women in the United States choosing a cervical barrier contraceptive method has dramatically declined. By 2002, fewer than 3% of women reported using any woman-initiated barrier method, including the diaphragm, female condom, or cervical cap. At the same time, however, research in infectious diseases indicates that cervical barriers may effectively prevent the transmission of several sexually transmitted infection. This possibility has fueled the recent development of two novel devices. This article examines the seven devices currently available in the United States, comparing their characteristics, efficacy, benefits, and drawbacks. Compared to the diaphragm, the new devices do not offer improved odds of pregnancy prevention, and evidence for their efficacy is sparse. Reasons for the limited acceptance of these methods as contraceptives on one hand-and for interest in their potential for limiting sexually transmitted infections on the other-will also be reviewed. Despite the limited acceptance of cervical barrier methods, midwives and other clinicians should promote their availability as an alternative to other reversible contraceptives. PMID- 17081940 TI - Providing quality care to the sexual assault survivor: education and training for medical professionals. AB - Health care providers who perform sexual assault examinations can assist patients who report experiencing a sexual assault by increasing their knowledge and skills in sexual assault care, and serving as Sexual Assault Response Team (SART) coordinators, or as SART team members, in their communities. With little additional material, this training could be provided as a component of basic midwifery and/or advanced practice nursing education programs. This article reviews the essential steps and required training for conducting sexual assault examinations. PMID- 17081941 TI - Women's Health Care Minimum Data Set: pilot test and validation for use in clinical practice. AB - Basic elements of the structure, process, and outcomes of midwifery practice have not been fully determined, particularly in the areas of women's gynecologic and primary health care. The American College of Nurse-Midwives (ACNM) supported the development of clinical data sets to describe structure, process, and outcomes of midwifery practice for use by clinical practitioners. The Woman's Health Care Minimum Data Set was developed using a panel of expert midwives and other women's health care professionals, as well as literature resources. Students of the Graduate Midwifery Program at Philadelphia University performed pilot testing of the Woman's Health Care Minimum Data Set as a service to the profession of midwifery while applying concepts learned in their research methods courses. Each student (n = 19) recruited a midwifery practice in which she had a clinical affiliation, and gathered data sets on the previous 30 consecutive women's health care encounters by CNMs or CMs (n = 569). Item analysis and refinement were done. Criterion-related validity and construct-related validity of the Woman's Health Care Minimum Data Set were explored through comparison with the medical record and through the testing of plausible hypotheses. The Woman's Health Care Minimum Data Set has the potential to be an important instrument in documenting and understanding the evolving nature of the practice of primary women's health care by midwives and other women's health care providers. PMID- 17081942 TI - Vulvar pain syndromes: vestibulodynia. AB - Chronic pain anywhere on the body can be debilitating and demoralizing. When the pain is associated with sexuality, it can erode self-esteem and diminish relationships. Vestibulodynia (pain in the vulvar vestibule) is poorly understood and presents a clinical challenge to the provider. Although the etiology of vestibulodynia is unclear, and randomized controlled trials of therapies are lacking, the knowledge of current theories and treatments will assist providers in caring for women with this enigmatic problem. PMID- 17081943 TI - Essure: hysteroscopic sterilization. AB - Essure, a new female sterilization method approved by the US Food and Drug Administration, offers women a new option for controlling fertility. The technique is done without an incision, is hormone-free, and can be performed on an outpatient basis under local anesthesia. There are few or no side effects. It is important for providers to understand how the device works, appropriate candidates for use, and requirements for preplacement and postplacement management. PMID- 17081944 TI - Clinical update: human papillomavirus vaccine. PMID- 17081948 TI - On the voyage of the beagle and other great adventures. PMID- 17081949 TI - Alternative sources of adult stem cells: a possible solution to the embryonic stem cell debate. AB - The complex moral and ethical debate surrounding the definition of the origins of human life, together with conflicting current and proposed legislation on state and federal levels, is hindering the course of research into the therapeutic uses of human embryonic stem cells. However, newly identified sources of adult stem cells, free from many of the ethical and legal concerns attached to embryonic stem cell research, may offer great promise for the advancement of medicine. These alternative sources may alleviate the need to resolve the stem cell debate before further therapeutic benefits of stem cell research can be realized. While legislation and ethics evolve to address the legal and moral issues of embryonic stem cell research, innovative researchers will continue to search for and find real and present solutions for cell-based therapies using adult stem cells. PMID- 17081950 TI - Gaps in knowledge in treating pregnant women. AB - Because studies are often undertaken without knowledge of the pharmacokinetics of a drug, efficacy is difficult to assess in pregnant women. To address this lack, basic and clinical research within the National Institute of Child Health and Human Development is focusing on expanding knowledge of pharmacology during pregnancy. Although medication use, including prescription, over-the-counter, and herbal products, is common during pregnancy, physicians may not be aware of the nonprescription products their patients are taking or the interactions these products may have with prescribed medications. A number of studies have found sex differences in oxidative metabolism and transport, as well as pharmacologic and toxicologic differences in hepatic metabolism, that are ultimately reflected in pharmacokinetics. Sex differences exist in distribution volumes, transport proteins, and drug clearance. Beyond these sex differences, pregnancy itself affects the absorption, distribution, metabolism, and elimination of a drug. Women experience more adverse drug reactions (ADRs) than do men, and these reactions tend to be more severe. QT prolongation (torsades de pointes) and hepatic toxicity are two of the most severe ADRs, frequently causing withdrawal of a drug from the market. Women may also metabolize drugs more quickly than do men, and drugs metabolized by cytochrome P3A4 are cleared more rapidly during pregnancy. A substantial increase in the clearance of drugs eliminated by renal mechanisms also has been noted. A significant number of women are clinically depressed during pregnancy and postpartum, and eliminating treatment for depression during pregnancy may have negative consequences for both mother and fetus. Among women with depression who are treated with selective serotonin reuptake inhibitors, the dose needed to maintain efficacy increases across the course of pregnancy. Drug disposition and response not only can differ between men and women, but also between pregnant and nonpregnant women. Research is needed to understand how pregnancy alters the pharmacokinetics and pharmacodynamics of drugs; then, efficacy trials can be initiated. Alternative strategies also need to be developed to characterize safety information. PMID- 17081951 TI - In pursuit of higher education, do we postpone parenthood too long? PMID- 17081952 TI - Attitudes toward parenthood and awareness of fertility among postgraduate students in Sweden. AB - BACKGROUND: Postponing childbirth is becoming increasingly common in Western countries, especially among women with higher education. OBJECTIVE: The aim of the present study was to investigate female and male postgraduate students' attitudes toward parenthood, their intentions to have children, perceived obstacles to having or raising children during postgraduate studies, and their awareness of fertility issues. METHODS: A 52-question survey was mailed to randomly selected postgraduate students attending a Swedish university. The questionnaire was developed based on earlier research by, and the professional experience of, the authors. Three pilot studies were conducted to test the questionnaire's reliability and validity. The t, x2, and Mann-Whitney U tests were used, as appropriate, to analyze responses. RESULTS: Two hundred female and 200 male postgraduate students agedC transition in exon 8 at position 25985 was identified, changing codon 217 from Leu to Pro (L217P). The mutation is located in the NADH binding domain at the base of alpha-helix Nalpha3, a region of sequence highly conserved from yeast to man. A quantitative assessment of the thermodynamic cost of this mutation at 37 degrees C revealed a ten-fold drop in the free energy of stability. Alterations in hydrogen bonding and solvent accessibility surrounding residue 217 were predicted based on computer modeling. PMID- 17082012 TI - Treatment-related myelodysplasia following fludarabine combination chemotherapy. AB - Although myelodysplasia (MDS) and secondary acute myeloid leukemia (sAML) are rare following fludarabine monotherapy, the risk of these diseases may potentially be increased when fludarabine is combined with cyclophosphamide or mitoxantrone due to synergistic effects on the inhibition of DNA repair. Among 137 patients treated with fludarabine combination regimens, ten patients developed MDS/sAML, including one who had received no other therapy. Six patients had abnormalities of chromosomes 5 and/or 7. The crude rate of MDS/sAML was 2.5% for previously untreated patients, and 9.3% for pretreated patients (p=0.28). The rate of MDS/sAML following fludarabine combination therapy is higher than that previously reported for fludarabine monotherapy. PMID- 17082013 TI - The outcome of autologous stem cell transplantation in patients with plasma cell disorders and dialysis-dependent renal failure. AB - Patients with multiple myeloma and end-stage renal failure on dialysis are frequently not considered eligible for high-dose therapy (HDT) due to higher transplant-related mortality (TRM). Our aim was to evaluate the toxicity and survival of dialysis-dependent patients after HDT with melphalan (100 mg/m(2)) compared to those of patients without renal insufficiency (melphalan 200 mg/m(2)) in a matched pairs analysis of 34 patients. No significant differences were observed between hematologic toxicity, TRM or disease response. Dialysis patients showed comparable event-free and overall survival. They required significantly extended intravenous antibiotic treatment and longer hospitalization. Thus, melphalan 100 mg/m2 is less toxic, yet equally efficient and improves the prognosis of this group of patients. PMID- 17082015 TI - The use of capillary blood samples in a large scale screening approach for the detection of beta-thalassemia and hemoglobin variants. AB - Hemoglobinopathies are priority genetic diseases for prevention programs in at risk populations. We implemented an accurate and simple methodology to identify hemoglobin (Hb) variants and to quantify HbA2 and HbF in capillary blood samples stored at room temperature for up to 7 days after collection. This methodology is particularly indicated for screening for carriers in primary care medical centers in which facilities for collecting venous blood are not available. PMID- 17082014 TI - Outcomes after myeloablative unrelated donor stem cell transplantation using both in vitro and in vivo T-cell depletion with alemtuzumab. AB - HLA-matched unrelated donor (MUD) stem cell transplantation (MUD) is complicated by a high incidence of graft-versus-host-disease (GVHD) resulting in significant morbidity and mortality. To circumvent this problem we included alemtuzumab for in vivo and in vitro T-cell depletion in a myeloablative MUD-SCT regimen. After SCT, no severe acute GVHD was observed in the 30 transplanted patients. Donor lymphocyte infusion administered at a later time point resulted in sustained anti tumor responses in most patients with chronic myeloid leukemia. After donor lymphocyte infusion three patients developed severe acute GVHD. Due to good responsiveness to immunosuppressive therapy only two patients developed persistent chronic GVHD. The main advantage of the transplantation regimen including alemtuzumab is that not only mortality due to GVHD is limited but also extensive chronic GVHD, which potentially leads to chronic morbidity and diminished quality of life, is hardly observed. PMID- 17082016 TI - Increased oxidative DNA products in patients with acute promyelocytic leukemia during arsenic therapy. AB - Arsenic trioxide (ATO) has been used to treat acute promyelocytic leukemia (APL), but the oxidative DNA damage occurring in patients has not been fully elucidated. We measured 8-hydroxy-2'-deoxyguanosine (8-OHdG), one of the most abundant oxidative products of DNA, by enzyme-linked immunoassay, and reactive oxidative species (ROS), by luminol- and luminol-H2O2 chemiluminescence, in the plasma of four APL patients treated with ATO. After six courses of ATO therapy, the plasma 8-OHdG concentration had increased from 45.6+/-22.8 ng/mL to 310.2+/-239.6 ng/mL. The plasma chemiluminescence level did not change significantly. These findings suggest that ATO generates intracellular oxidative DNA damage, but this is not correlated with the plasma ROS level. The clinical significance of 8-OHdG during and after ATO therapy warrants further study. PMID- 17082163 TI - Ethical issues in research with patients who have experienced stroke. AB - There are a myriad of ethical issues surrounding inclusion of persons who have had strokes into clinical research. Assessment of decision-making capacity is especially challenging as stroke can impair communication such that the patient appears to lack decision-making capacity or can impair executive function and preserve expressive language so that it appears the patient has capacity when he or she may not. Other issues such as vulnerability, therapeutic misconception, surrogate decision making, and other concerns unique to persons who have had strokes are discussed. Recommendations of national commissions are described. PMID- 17082164 TI - Ethics, family caregivers, and stroke. AB - Even though stroke caregivers provide valuable care to loved ones, often sacrificing their own life goals and sometimes their health, they are frequently viewed by stroke professionals as irritants or problems. Medical training and medical bioethics literature focus almost exclusively on the physician-patient relationship, which leaves family members with uncertain standing. A family centered approach allows providers to acknowledge and respect caregivers' needs without sacrificing patients' interests or authority. PMID- 17082165 TI - Ethical issues in the management of dysphagia after stroke. AB - When patients have severe dysphagia after a stroke, tube feeding may be recommended to reduce the risks associated with malnutrition, dehydration, and/or aspiration. Patients may not be able to participate in decision making, but they may have previously expressed strong preferences related to tube feeding. Clinicians must work together with the family to establish a treatment plan that is respectful of the person's previous wishes, yet mindful of the flaws in advance care planning. Although ethical issues cannot be avoided, clinicians can reduce uncertainty by understanding current ethical and legal views on these challenging issues. PMID- 17082166 TI - The cognitive, emotional, and social sequelae of stroke: psychological and ethical concerns in post-stroke adaptation. AB - In the clinical domain, ethical analyses involve examination of complex individual responses, psychological processes, and social context. Psychological aspects of stroke adaptation include the risk for depression and anxiety, changes in identity and personality processes, and potential for social isolation. Depression and anxiety are heterogeneous constructs and can affect individuals' emotional functioning and cognitive abilities. Executive function, self-agency, and volition may be affected. Alterations in identity and personality may also result from the interaction of fluctuating emotional, cognitive, and physical abilities as well as from changes in social context and family dynamics. Social isolation, or lack of access to social contact or resources, can be a consequence of difficulties in cognitive and emotional function that influence interpersonal relationships, changes in social roles, communication difficulties, and challenges in transportation and employment. Social stigma and marginalization also contribute to isolation. The authors describe these psychological phenomena in the context of brain damage and recovery and raise ethical concerns including impact on decision-making capacity, pre- and postinjury selves and interests, and the social milieu in which strokes are experienced. PMID- 17082167 TI - Failure to achieve assent in a communicative patient: what are the caregiver's obligations? AB - Disclosure of facts related to an individual's health care, including level and location of care, are central to the fulfillment of informed consent. When an individual is unable to provide informed consent because they lack the decision making capacity to process the information and/or they have been deemed legally incompetent, the health care team should obtain assent from the individual whenever possible. Questions remain as to how to proceed when assent cannot be gained and the decision involved causes a level of distress for the individual that is difficult to resolve. A focus on maintaining hope and support with a clear transition and follow-up plan are important variables to consider for an ethical, caring response. PMID- 17082168 TI - Is informed consent a "yes or no" response? Enhancing the shared decision-making process for persons with aphasia. AB - Respect for patient autonomy and the need to have a comprehensive discussion of the risks and benefits of a medical intervention are two important issues involved in the process of obtaining informed consent. In dealing with individuals with aphasia, there may be particular challenges in balancing these two ethical imperatives. Although decision-making capacity may be preserved with aphasia, the patients' ability to fully participate in a dialogue regarding a proposed medical intervention is frequently impaired. We propose a process of enhancing informed consent for persons with aphasia while continuing to respect and enhance patient autonomy and the exercise of decision making for these patients. The use of a patient-selected "helper" during the informed consent process can improve the quality of the informed consent, while reserving final decision-making authority for the patient. PMID- 17082169 TI - Ethical issues involving the right hemisphere stroke patient: to treat or not to treat? AB - The management of patients with right hemisphere damage (RHD) presents a challenge to the allied health clinician. In addition to impairments in specific cognitive areas, some patients may not be aware of the presence or extent of their deficits or the impact of these deficits on everyday activities. This article briefly describes two related models of awareness/unawareness that may help guide the assessment and treatment process in patients with neurological damage including RHD. Then, some clinical care decisions regarding patients with decreased awareness are presented together with a framework for organizing and evaluating the various factors associated with a case. PMID- 17082170 TI - Complexities in decision making for persons with disabilities nearing end of life. AB - Good end-of-life care requires that clinicians, families, and ethicists be aware of biases that influence patient cases, particularly in the acute care setting where the aim is primarily cure and return to optimal functional level. Persons with disabilities may pose unique challenges; their potential for quality of life is viewed through the lens of highly functional clinicians who might have a biased view of the disabled person's quality of life. The authors aim to present three categories of disability that do not claim to be absolute but rather offer clinicians and ethicists a lens through which to reflect on bias that unconsciously may influence their approach to the patient who is seriously ill and may be nearing the end of life. The categories include (a) a person who has lived with a disability from birth or early life, due to trauma or disease, and is now faced with a serious illness that requires that life-sustaining treatment; (b) the otherwise healthy person who acquires a disability through an acute event of disease or trauma and whose condition requires that life-sustaining treatment decisions be made; and (c) the person who has lived with a progressive chronic illness, such as lung or heart disease or amyotrophic lateral sclerosis, and may have gradually adjusted to disabilities imposed by the condition and now is faced with life-sustaining treatment decisions. The concept of inherent dignity (Pellegrino 2005) is suggested as a filtering lens in case consideration. PMID- 17082171 TI - Abandoning the Golden Rule: the problem with "putting ourselves in the patient's place". AB - A large body of evidence documents the difficulties health care professionals have in predicting their patient's beliefs or wishes. These difficulties extend from the predictions of very specific patient wishes (such as for life-sustaining therapies) to more global assessments of patients' lives as a whole (for instance, their quality of life). Although many explanations have been offered for this phenomenon, we discuss one that has not received as much attention: the conscious or unconscious adoption of what we refer to as Golden Rule thinking. This refers to our attempts to understand another person's situation by imagining what we would believe or want under similar circumstances, in other words, "putting ourselves in the patient's place." Although Golden Rule thinking would seem to be a promising strategy, studies show that it actually results in inaccurate predictions of patients' wishes or beliefs. These mispredictions, in turn, have significant clinical and ethical implications. We review possible reasons why Golden Rule thinking may be of limited utility in understanding our patients' situations and suggest alternate strategies to maximize our understanding of our patient's lives. PMID- 17082172 TI - Consensus panel guidelines for the use of intrathecal baclofen therapy in poststroke spastic hypertonia. AB - Intrathecal baclofen (ITB) therapy has been increasingly employed for the management of poststroke spastic hypertonia, a complication that can lead to deformity, discomfort, and exacerbation of motor impairments. Because its use in stroke is not as established as other indications, ITB therapy has not been subjected to rigorous investigation. There is limited evidence to guide clinicians regarding application of this therapy in this patient population. This article aims to review the available scientific literature and the opinion of several experts on the topic. It will also describe the recommendations of these experts with regard to addressing common clinical situations that may influence treatment decisions in the stroke population. PMID- 17082173 TI - Long-term changes in participation after stroke. AB - BACKGROUND AND PURPOSE: People who have had a stroke may have difficulty resuming some of their previous activities, which leads to a decline in their participation in daily activities and social roles. The purposes of this study were to compare participation 6 months (T1) and between 2 and 4 years (T2) after discharge from a rehabilitation unit and to verify if any changes were associated with changes in personal and environmental factors. METHOD: Participation of people who had had a stroke was measured at T1 and T2 with the Assessment of Life Habits. RESULTS: A significant reduction (p < .001) in participation in daily activities was observed, specifically in the following categories: nutrition, p < .001; fitness, p = .004; personal care, p < .001; and housing, p = .001. However, participation in social roles was maintained during this period (p = .10). The increased perception of technology as a facilitator (environmental factor) over time explained a part of the decline in participation (R2 = 0.13). CONCLUSION: Factors associated with the reduction in participation in daily activities should be further studied in order to prevent this decline. PMID- 17082174 TI - Stroke care in long-term care facilities in southwestern Ontario. AB - PURPOSE: This study describes the current state of stroke care and rehabilitation in facility-based long-term care (LTC). METHOD: LTC representatives, community partners (including physiotherapists and occupational therapists), stroke survivors, and family members were interviewed about stroke care provided in LTC facilities. RESULTS: Limitations of the current system were identified including inadequate provision of therapy, unequal access to specialized rehabilitation, lack of staff training to care for the specialized needs of residents with stroke, and the lack of coordinated and consistent care. CONCLUSION: This study identified challenges and barriers to providing optimal stroke care in LTC facilities. Recommendations for enhancing stroke care are suggested. PMID- 17082175 TI - Expression profiling defines a recurrence signature in lung squamous cell carcinoma. AB - Lung cancer remains the leading cause of cancer death worldwide. Overall 5-year survival is approximately 10-15% and despite curative intent surgery, treatment failure is primarily due to recurrent disease. Conventional prognostic markers are unable to determine which patients with completely resected disease within each stage group are likely to relapse. To identify a gene signature associated with recurrent squamous cell carcinoma (SCC) of lung, we analyzed primary tumor gene expression for a total of 51 SCCs (Stages I-III) on 22 323 element microarrays, comparing expression profiles for individuals who remained disease free for a minimum of 36 months with those from individuals whose disease recurred within 18 months of complete resection. Cox proportional hazards modeling with leave-one-out cross-validation identified a 71-gene signature capable of predicting the likelihood of tumor recurrence and a 79-gene signature predictive for cancer-related death. These two signatures were pooled to generate a 111-gene signature which achieved an overall predictive accuracy for disease recurrence of 72% (77% sensitivity, 67% specificity) in an independent set of 58 (Stages I-III SCCs). This signature also predicted differences in survival [log rank P=0.0008; hazard ratio (HR), 3.8; 95% confidence interval (CI), 1.6-8.7], and was superior to conventional prognostic markers such as TNM stage or N stage in predicting patient outcome. Genome-wide profiling has revealed a distinct gene expression profile for recurrent lung SCC which may be clinically useful as a prognostic tool. PMID- 17082176 TI - Role of NQO1C609T and EPHX1 gene polymorphisms in the association of smoking and alcohol with sporadic distal colorectal adenomas: results from the UKFSS Study. AB - NADP(H):quinone oxidoreductase 1 (NQO1) and microsomal epoxide hydrolase (EPHX1, also mEH) are attractive candidate enzymes for association with colorectal neoplasia because they metabolize a number of compounds including polycyclic aromatic hydrocarbons (PAHs) that have been linked with colorectal carcinogenesis. We examined the relationship between NQO1C609T, mEH3, mEH4 and risk of sporadic distal colorectal adenomas in one of the largest case-control studies of 946 polyp-free controls and 894 cases, all participants of the UK Flexible Sigmoidoscopy Screening (UKFSS) Trial. The polymorphisms were examined as independent risk factors and evidence for interaction with smoking and alcoholic drinks was sought. The NQO1 609*T allele was positively associated with high-risk adenoma in this population [odds ratio (OR), 1.36; 95% confidence interval (CI), 1.02-1.83]. Elevated risk estimates were seen in smokers independently of the genotype but the association was stronger among current smokers with the heterozygous variant genotype (OR, 4.24; 95% CI, 2.54-7.09). It was reported for the first time that the association between alcohol and colorectal adenoma was modified by NQO1C609T genotype, such that the relation between alcohol and colorectal adenoma was stronger among those with the common C/C genotype (OR, 1.49; 95% CI, 1.11-2.02; P-interaction = 0.024). There was no association between mEH3 and mEH4 variants and colorectal adenoma risk and no effect modification by alcohol and smoking. These findings provide evidence for an important role of the NQO1C609T polymorphism in susceptibility of colorectal adenomas. Alcohol increases risk of colorectal adenoma in carriers of the high activity genotype possibly through enhanced activation of alcohol-related procarcinogens. PMID- 17082177 TI - A meat and potato war: implications for cancer etiology. AB - Cooking foods clearly has a beneficial impact for humans; the microbial content can be decreased, proteins made more digestible and the flavor and texture improved. But at the same time, amino acids, creatine and sugars, which occur naturally in meats, may be involved in reactions that generate heterocyclic amine (HA) carcinogens during cooking. Recently, another amine carcinogen, acrylamide, was found at relatively high levels in cooked carbohydrate-rich foods, especially potatoes. In this commentary acrylamide will be compared with the meat carcinogens (HAs) with respect to formation, human intake and health consequences -it's a meat and potato war. What conclusion about risks from these dietary carcinogens can we make from the available scientific data? PMID- 17082178 TI - Effect of pseudophosphorylation and cross-linking by lipid peroxidation and advanced glycation end product precursors on tau aggregation and filament formation. AB - Accumulation of hyperphosphorylated Tau protein as paired helical filaments in pyramidal neurons is a major hallmark of Alzheimer disease. Besides hyperphosphorylation, other modifications of the Tau protein, such as cross linking, are likely to contribute to the characteristic features of paired helical filaments, including their insolubility and resistance against proteolytic degradation. In this study, we have investigated whether the four reactive carbonyl compounds acrolein, malondialdehyde, glyoxal, and methylglyoxal accelerate the formation of Tau oligomers, thioflavin T-positive aggregates, and fibrils using wild-type and seven pseudophosphorylated mutant Tau proteins. Acrolein and methylglyoxal were the most reactive compounds followed by glyoxal and malondialdehyde in terms of formation of Tau dimers and higher molecular weight oligomers. Furthermore, acrolein and methylglyoxal induced the formation of thioflavin T-fluorescent aggregates in a triple pseudophosphorylation mimicking mutant to a slightly higher degree than wild-type Tau. Analysis of the Tau aggregates by electron microscopy study showed that formation of fibrils using wild-type Tau and several Tau mutants could be observed with acrolein and methylglyoxal but not with glyoxal and malondialdehyde. Our results suggest that reactive carbonyl compounds, particularly methylglyoxal and acrolein, could accelerate tangle formation in vivo and that this process could be slightly accelerated, at least in the case of methylglyoxal and acrolein, by hyperphosphorylation. Interference with the formation or the reaction of these reactive carbonyl compounds could be a promising way of inhibiting tangle formation and neuronal dysfunction in Alzheimer disease and other tauopathies. PMID- 17082179 TI - BCL2 is a downstream effector of MIZ-1 essential for blocking c-MYC-induced apoptosis. AB - The c-MYC oncoprotein is among the most potent transforming agents in human cells. Ironically, c-MYC is also capable of inducing massive apoptosis under certain conditions. A clear understanding of the distinct pathways activated by c MYC during apoptosis induction and transformation is crucial to the design of therapeutic strategies aimed at selectively reactivating the apoptotic potential of c-MYC in cancer cells. We recently demonstrated that apoptosis induction in primary human cells strictly requires that c-MYC bind and inactivate the transcription factor MIZ-1. This presumably blocked the ability of MIZ-1 to activate the transcription of an unidentified pro-survival gene. Here we report that MIZ-1 activates the transcription of BCL2. More importantly, inhibition of the MIZ-1/BCL2 signal is an essential event during the apoptotic response. Furthermore, targeting BCL2 with short hairpin RNA or small molecule inhibitors restores the apoptotic potential of a c-MYC mutant that is defective for MIZ-1 inhibition. These observations suggest that repression of BCL2 transcription is the single essential consequence of targeting the MIZ-1 pathway during apoptosis induction. These data define a genetic pathway that helps to explain historical observations documenting cooperation between c-MYC and BCL2 overexpression in human cancer. PMID- 17082180 TI - Structural determinants of human FANCF protein that function in the assembly of a DNA damage signaling complex. AB - Fanconi anemia (FA) is a rare autosomal recessive and X-linked chromosomal instability disorder. At least eight FA proteins (FANCA, B, C, E, F, G, L, and M) form a nuclear core complex required for monoubiquitination of a downstream protein, FANCD2. The human FANCF protein reportedly functions as a molecular adaptor within the FA nuclear complex, bridging between the subcomplexes A:G and C:E. Our x-ray crystallographic studies of the C-terminal domain of FANCF reveal a helical repeat structure similar to the Cand1 regulator of the Cul1-Rbx1-Skp1 Fbox(Skp2) ubiquitin ligase complex. Two C-terminal loops of FANCF are essential for monoubiquitination of FANCD2 and normal cellular resistance to the DNA cross linking agent mitomycin C. FANCF mutants bearing amino acid substitutions in this C-terminal surface fail to interact with other components of the FA complex, indicating that this surface is critical for the proper assembly of the FA core complex. PMID- 17082181 TI - Epidermal growth factor receptor (EGFR) antibody down-regulates mutant receptors and inhibits tumors expressing EGFR mutations. AB - Activating mutations in the kinase domain of the EGF receptor have been reported in non-small cell lung cancer. The majority of tumors expressing these mutants are sensitive to ATP mimetics that inhibit the EGFR tyrosine kinase. The effect of antibodies that bind to the ectodomain of the receptor is less clear. We report herein the effects and mechanisms of action of the antibody cetuximab in lung cancer cells that naturally express receptor mutations and in ErbB-null 32D hematopoietic cells transfected with mutant EGFR. Treatment with cetuximab down regulated EGFR levels and inhibited cell growth both in vitro and in vivo. This was associated with inhibition of ligand-independent EGFR signaling. These effects were seen in 32D cells arguing the growth inhibitory action was not because of the blockade of autocrine ligand action. Both antibody-induced EGFR down-regulation and inhibition of growth required receptor dimerization as monovalent Fab fragments only eliminated receptor levels or reduced cell proliferation in the presence of antihuman IgG. Finally, cetuximab inhibited growth of H1975 lung cancer cells and xenografts, which expressed L858R/T790M EGFR and were resistant to EGFR tyrosine kinase inhibitors. These data suggest that cetuximab is an effective therapy against mutant EGFR-expressing cancer cells and thus can be considered in combination with other anti-EGFR molecules. PMID- 17082182 TI - Structure of human spindlin1. Tandem tudor-like domains for cell cycle regulation. AB - Spindlin1, a meiotic spindle-binding protein that is highly expressed in ovarian cancer cells, was first identified as a gene involved in gametogenesis. It appeared to be a target for cell cycle-dependent phosphorylation and was demonstrated to disturb the cell cycle. Here we report the crystal structure of human spindlin1 to 2.2A of resolution, representing the first three-dimensional structure from the spin/ssty (Y-linked spermiogenesis-specific transcript) gene family. The refined structure, containing three repeats of five/four anti parallel beta-strands, exhibits a novel arrangement of tandem Tudor-like domains. Two phosphate ions, chelated by Thr-95 and other residues, appear to stabilize the long loop between domains I and II, which might mediate the cell cycle regulation activity of spindlin1. Flow cytometry experiments indicate that cells expressing spindlin1 display a different cell cycle distribution in mitosis, whereas those expressing a T95A mutant, which had a great decrease in phosphorous content, have little effect on the cell cycle. We further identified associations of spindlin1 with nucleic acid to provide a biochemical basis for its cell cycle regulation and other functions. PMID- 17082183 TI - Evidence for the pathophysiological role of endogenous methylarginines in regulation of endothelial NO production and vascular function. AB - In endothelium, NO is derived from endothelial NO synthase (eNOS)-mediated L arginine oxidation. Endogenous guanidinomethylated arginines (MAs), including asymmetric dimethylarginine (ADMA) and NG-methyl-L-arginine (L-NMMA), are released in cells upon protein degradation and are competitive inhibitors of eNOS. However, it is unknown whether intracellular MA concentrations reach levels sufficient to regulate endothelial NO production. Therefore, the dose-dependent effects of ADMA and L-NMMA on eNOS function were determined. Kinetic studies demonstrated that the Km for L-arginine is 3.14 microM with a Vmax of 0.14 micromol mg-1 min-1, whereas Ki values of 0.9 microM and 1.1 microM were determined for ADMA and L-NMMA, respectively. EPR studies of NO production from purified eNOS demonstrated that, with a physiological 100 microM level of L arginine, MA levels of >10 microM were required for significant eNOS inhibition. Dose-dependent inhibition of NO formation in endothelial cells was observed with extracellular MA concentrations as low 5 microm. Similar effects were observed in isolated vessels where 5 microm ADMA inhibited vascular relaxation to acetylcholine. MA uptake studies demonstrated that ADMA and L-NMMA accumulate in endothelial cells with intracellular levels greatly exceeding extracellular concentrations. L-arginine/MA ratios were correlated with cellular NO production. Although normal physiological levels of MAs do not significantly inhibit NOS, a 3 to 9-fold increase, as reported under disease conditions, would exert prominent inhibition. Using a balloon model of vascular injury, approximately 4-fold increases in cellular MAs were observed, and these caused prominent impairment of vascular relaxation. Thus, MAs are critical mediators of vascular dysfunction following vascular injury. PMID- 17082184 TI - Deficiency in phylloquinone (vitamin K1) methylation affects prenyl quinone distribution, photosystem I abundance, and anthocyanin accumulation in the Arabidopsis AtmenG mutant. AB - Phylloquinone (vitamin K(1)) is synthesized in cyanobacteria and in chloroplasts of plants, where it serves as electron carrier of photosystem I. The last step of phylloquinone synthesis in cyanobacteria is the methylation of 2-phytyl-1,4 naphthoquinone by the menG gene product. Here, we report that the uncharacterized Arabidopsis gene At1g23360, which shows sequence similarity to menG, functionally complements the Synechocystis menG mutant. An Arabidopsis mutant, AtmenG, carrying a T-DNA insertion in the gene At1g23360 is devoid of phylloquinone, but contains an increased amount of 2-phytyl-1,4-naphthoquinone. Phylloquinone and 2 phytyl-1,4-naphthoquinone in thylakoid membranes of wild type and AtmenG, respectively, predominantly localize to photosystem I, whereas excess amounts of prenyl quinones are stored in plastoglobules. Photosystem I reaction centers are decreased in AtmenG plants under high light, as revealed by immunoblot and spectroscopic measurements. Anthocyanin accumulation and chalcone synthase (CHS1) transcription are affected during high light exposure, indicating that alterations in photosynthesis in AtmenG affect gene expression in the nucleus. Photosystem II quantum yield is decreased under high light. Therefore, the loss of phylloquinone methylation affects photosystem I stability or turnover, and the limitation in functional photosystem I complexes results in overreduction of photosystem II under high light. PMID- 17082185 TI - Alix facilitates the interaction between c-Cbl and platelet-derived growth factor beta-receptor and thereby modulates receptor down-regulation. AB - Alix (ALG-2-interacting protein X) is an adaptor protein involved in down regulation and sorting of cell surface receptors through the endosomal compartments toward the lysosome. In this study, we show that Alix interacts with the C-terminal region of the platelet-derived growth factor (PDGF) beta-receptor (PDGFRbeta) and becomes transiently tyrosine-phosphorylated in response to PDGF BB stimulation. Increased expression levels of Alix resulted in a reduced rate of PDGFRbeta removal from the cell surface following receptor activation, and this was associated with decreased receptor degradation. Furthermore, Alix was found to co-immunoprecipitate with the ubiquitin ligase c-Cbl, and elevated Alix levels increased the interaction between c-Cbl and PDGFRbeta. Interestingly, Alix interacted constitutively with both c-Cbl and PDGFRbeta. Moreover, c-Cbl was found to be hyperphosphorylated in cells engineered to overexpress Alix compared with control cells. The increased c-Cbl phosphorylation correlated with enhanced proteasomal degradation of c-Cbl, which in turn correlated with a decreased ubiquitination of PDGFRbeta. Our data suggest that Alix inhibits down-regulation of PDGFRbeta by modulating the interaction between c-Cbl and the receptor, thereby affecting the ubiquitination of the receptor. PMID- 17082186 TI - Biochemical basis of glucokinase activation and the regulation by glucokinase regulatory protein in naturally occurring mutations. AB - Glucokinase (GK) has several known polymorphic activating mutations that increase the enzyme activity by enhancing glucose binding affinity and/or by alleviating the inhibition of glucokinase regulatory protein (GKRP), a key regulator of GK activity in the liver. Kinetic studies were undertaken to better understand the effect of these mutations on the enzyme mechanism of GK activation and GKRP regulation and to relate the enzyme properties to the associated clinical phenotype of hypoglycemia. Similar to wild type GK, the transient kinetics of glucose binding for activating mutations follows a general two-step mechanism, the formation of an enzyme-glucose complex followed by an enzyme conformational change. However, the kinetics for each step differed from wild type GK and could be grouped into specific types of kinetic changes. Mutations T65I, Y214C, and A456V accelerate glucose binding to the apoenzyme form, whereas W99R, Y214C, and V455M facilitate enzyme isomerization to the active form. Mutations that significantly enhance the glucose binding to the apoenzyme also disrupt the protein-protein interaction with GKRP to a large extent, suggesting these mutations may adopt a more compact conformation in the apoenzyme favorable for glucose binding. Y214C is the most active mutation (11-fold increase in k(cat)/K(0.5)(h)) and exhibits the most severe clinical effects of hypoglycemia. In contrast, moderate activating mutation A456V nearly abolishes the GKRP inhibition (76-fold increase in K(i)) but causes only mild hypoglycemia. This suggests that the alteration in GK enzyme activity may have a more profound biological impact than the alleviation of GKRP inhibition. PMID- 17082187 TI - Sordarin derivatives induce a novel conformation of the yeast ribosome translocation factor eEF2. AB - The sordarins are fungal specific inhibitors of the translation factor eEF2, which catalyzes the translocation of tRNA and mRNA after peptide bond formation. We have determined the crystal structures of eEF2 in complex with two novel sordarin derivatives. In both structures, the three domains of eEF2 that form the ligand-binding pocket are oriented in a different manner relative to the rest of eEF2 compared with our previous structure of eEF2 in complex with the parent natural product sordarin. Yeast eEF2 is also shown to bind adenylic nucleotides, which can be displaced by sordarin, suggesting that ADP or ATP also bind to the three C-terminal domains of eEF2. Fusidic acid is a universal inhibitor of translation that targets EF-G or eEF2 and is widely used as an antibiotic against Gram-positive bacteria. Based on mutations conferring resistance to fusidic acid, cryo-EM reconstructions, and x-ray structures of eEF2, EF-G, and an EF-G homolog, we suggest that the conformation of EF-G stalled on the 70 S ribosome by fusidic acid is similar to that of eEF2 trapped on the 80 S ribosome by sordarin. PMID- 17082188 TI - Structural reorganization and the cooperative binding of single-stranded telomere DNA in Sterkiella nova. AB - In Sterkiella nova, alpha and beta telomere proteins bind cooperatively with single-stranded DNA to form a ternary alpha.beta.DNA complex. Association of telomere protein subunits is DNA-dependent, and alpha-beta association enhances DNA affinity. To further understand the molecular basis for binding cooperativity, we characterized several possible stepwise assembly pathways using isothermal titration calorimetry. In one path, alpha and DNA first form a stable alpha.DNA complex followed by the addition of beta in a second step. Binding energy accumulates with nearly equal free energy of association for each of these steps. Heat capacity is nonetheless dramatically different, with DeltaCp = -305 +/- 3 cal mol(-1) K(-1) for alpha binding with DNA and DeltaCp = -2010 +/- 20 cal mol(-1) K(-1) for the addition of beta to complete the alpha.beta.DNA complex. By examining alternate routes including titration of single-stranded DNA with a preformed alpha.beta complex, a significant portion of binding energy and heat capacity could be assigned to structural reorganization involving protein-protein interactions and repositioning of the DNA. Structural reorganization probably affords a mechanism to regulate high affinity binding of telomere single-stranded DNA with important implications for telomere biology. Regulation of telomere complex dissociation is thought to involve post-translational modifications in the lysine-rich C-terminal portion of beta. We observed no difference in binding energetics or crystal structure when comparing complexes prepared with full length beta or a C-terminally truncated form, supporting interesting parallels between the intrinsically disordered regions of histones and this portion of beta. PMID- 17082189 TI - YB-1 is important for an early stage embryonic development: neural tube formation and cell proliferation. AB - The eukaryotic Y-box-binding protein-1 (YB-1) is involved in the transcriptional and translational control of many biological processes, including cell proliferation. In clinical studies, the cellular level of YB-1 closely correlates with tumor growth and prognosis. To understand the role of YB-1 in vivo, especially in the developmental process, we generated YB-1 knock-out mice, which are embryonic lethal and exhibit exencephaly associated with abnormal patterns of cell proliferation within the neuroepithelium. beta-Actin expression and F-actin formation were reduced in the YB-1 null embryo and YB-1(-/-) mouse embryonic fibroblasts, suggesting that the neural tube defect is caused by abnormal cell morphology and actin assembly within the neuroepithelium. Fibroblasts derived from YB-1(-/-) embryos demonstrated reduced growth and cell density. A colony formation assay showed that YB-1(-/-) mouse embryonic fibroblasts failed to undergo morphological transformation and remained contact-inhibited in culture. These results demonstrate that YB-1 is involved in early mouse development, including neural tube closure and cell proliferation. PMID- 17082190 TI - Ca2+-independent phospholipase A2-dependent gating of TRPM8 by lysophospholipids. AB - TRPM8 represents an ion channel activated by cold temperatures and cooling agents, such as menthol, that underlies the cold-induced excitation of sensory neurons. Interestingly, the only human tissue outside the peripheral nervous system, in which the expression of TRPM8 transcripts has been detected at high levels, is the prostate, a tissue not exposed to any essential temperature variations. Here we show that the TRPM8 cloned from human prostate and heterologously expressed in HEK-293 cells is regulated by the Ca(2+)-independent phospholipase A(2) (iPLA(2)) signaling pathway with its end products, lysophospholipids (LPLs), acting as its endogenous ligands. LPLs induce prominent prolongation of TRPM8 channel openings that are hardly detectable with other stimuli (e.g. cold, menthol, and depolarization) and that account for more than 90% of the total channel open time. Down-regulation of iPLA(2) resulted in a strong inhibition of TRPM8-mediated functional responses and abolished channel activation. The action of LPLs on TRPM8 channels involved either changes in the local lipid bilayer tension or interaction with the critical determinant(s) in the transmembrane channel core. Based on this, we propose a novel concept of TRPM8 regulation with the involvement of iPLA(2) stimulation. This mechanism employs chemical rather than physical (temperature change) signaling and thus may be the main regulator of TRPM8 activation in organs not exposed to any essential temperature variations, as in the prostate gland. PMID- 17082191 TI - Visualization of galectin-3 oligomerization on the surface of neutrophils and endothelial cells using fluorescence resonance energy transfer. AB - Galectin-3, a member of the galectin family of carbohydrate binding proteins, is widely expressed, particularly in cells involved in the immune response. Galectin 3 has also been indicated to play a role in various biological activities ranging from cell repression to cell activation and adhesion and has, thus, been recognized as an immunomodulator. Whereas those activities are likely to be associated with ligand cross-linking by this lectin, galectin-3, unlike other members of the galectin family, exists as a monomer. It has consequently been proposed that oligomerization of the N-terminal domains of galectin-3 molecules, after ligand binding by the C-terminal domain, is responsible for this cross linking. The oligomerization status of galectin-3 could, thus, control the majority of its extracellular activities. However, little is known about the actual mode of action through which galectin-3 exerts its function. In this report we present data suggesting that oligomerization of galectin-3 molecules occurs on cell surfaces with physiological concentrations of the lectin. Using galectin-3 labeled at the C terminus with Alexa 488 or Alexa 555, the oligomerization between galectin-3 molecules on cell surfaces was detected using fluorescence resonance energy transfer. We observed this fluorescence resonance energy transfer signal in different biological settings representing the different modes of action of galectin-3 that we previously proposed; that is, ligand crosslinking leading to cell activation, cell-cell interaction/adhesion, and lattice formation. Furthermore, our data suggest that galectin-3 lattices are robust and could, thus, be involved, as previously proposed, in the restriction of receptor clustering. PMID- 17082192 TI - Molecular recognition in the assembly of collagens: terminal noncollagenous domains are key recognition modules in the formation of triple helical protomers. AB - The alpha-chains of the collagen superfamily are encoded with information that specifies self-assembly into fibrils, microfibrils, and networks that have diverse functions in the extracellular matrix. A key self-organizing step, common to all collagen types, is trimerization that selects, binds, and registers cognate alpha-chains for assembly of triple helical protomers that subsequently oligomerize into specific suprastructures. In this article, we review recent findings on the mechanism of chain selection and infer that terminal noncollagenous domains function as recognition modules in trimerization and are therefore key determinants of specificity in the assembly of suprastructures. This mechanism is also illustrated with computer-generated animations. PMID- 17082193 TI - Skp2 controls adipocyte proliferation during the development of obesity. AB - The increase in the mass of adipose tissue during the development of obesity can arise through an increase in cell size, an increase in cell number, or both. Here we show that long term maintenance of C57BL/6 mice on a high fat diet (for approximately 25 weeks) induces an initial increase in adipocyte size followed by an increase in adipocyte number in white adipose tissue. The latter effect was found to be accompanied by up-regulation of expression of the gene for the F-box protein Skp2 as well as by downregulation of the cyclin-dependent kinase inhibitor p27(Kip1), a principal target of the SCF(Skp2) ubiquitin ligase, in white adipose tissue. Ablation of Skp2 protected mice from the development of obesity induced either by a high fat diet or by the lethal yellow agouti (A(y)) mutation, and this protective action was due to inhibition of the increase in adipocyte number without an effect on adipocyte hypertrophy. The reduction in the number of adipocyte caused by Skp2 ablation also inhibited the development of obesity-related insulin resistance in the A(y) mutant mice, although the reduced number of beta cells and reduced level of insulin secretion in Skp2-deficient mice resulted in glucose intolerance. Our observations thus indicate that Skp2 controls adipocyte proliferation during the development of obesity. PMID- 17082194 TI - The enzymatic function of tafazzin. AB - Tafazzin is a putative enzyme that is involved in cardiolipin metabolism, it may carry mutations responsible for Barth syndrome. To identify the biochemical reaction catalyzed by tafazzin, we expressed the full-length isoform of Drosophila melanogaster tafazzin in a baculovirus-Sf9 insect cell system. Tafazzin expression induced a new enzymatic function in Sf9 cell mitochondria, namely 1-palmitoyl-2-[14C]linoleoyl-phosphatidylcholine:monolysocardiolipin linoleoyltransferase. We also found evidence for the reverse reaction, because tafazzin expression caused transfer of acyl groups from phospholipids to 1 [14C]palmitoyl-2-lyso-phosphatidylcholine. An affinity-purified tafazzin construct, tagged with the maltose-binding protein, catalyzed both forward and reverse transacylations between cardiolipin and phosphatidylcholine, but was unable to utilize CoA or acyl-CoA as substrates. Whereas tafazzin supported transacylations between various phospholipid-lysophospholipid pairs, it showed the highest rate for the phosphatidylcholine-cardiolipin transacylation. Transacylation activities were about 10-fold higher for linoleoyl groups than for oleoyl groups, and they were negligible for arachidonoyl groups. The data show that Drosophila tafazzin is a CoA-independent, acyl-specific phospholipid transacylase with substrate preference for cardiolipin and phosphatidylcholine. PMID- 17082195 TI - Mechanical unfolding pathways of the enhanced yellow fluorescent protein revealed by single molecule force spectroscopy. AB - We used single molecule force spectroscopy to characterize the mechanical stability of the enhanced yellow fluorescent protein (EYFP) (a mutant form of the green fluorescent protein (GFP)) and two of its circularly permutated variants. In all three constructs, we found two main unfolding peaks; the first corresponds to a transition state placed close to the termini and the second to a transition state placed halfway through the molecule. We attribute the second transition state to the shear rupture of the beta1- and beta6-strands, which we verified by introducing a point mutation in this region. Although both unfolding peaks were observed in all three EYFP variants, their relative frequency of occurrence varied. Our results demonstrated that the mechanical unfolding pathways in EYFP could be deciphered through the use of circular permutation. PMID- 17082196 TI - AMP-activated protein kinase impairs endothelial actin cytoskeleton assembly by phosphorylating vasodilator-stimulated phosphoprotein. AB - Vasodilator-stimulated phosphoprotein (VASP) is an actin regulatory protein that links signaling pathways to remodeling of the cytoskeleton. VASP functions are modulated by protein kinases, which phosphorylate the sites Ser-157, Ser-239, and Thr-278. The kinase responsible for Thr-278 phosphorylation, biological functions of the phosphorylation, and association with disease states have remained enigmatic. Using VASP phosphorylation status-specific antibodies, we identified AMP-activated protein kinase (AMPK), a serine-threonine kinase and fundamental sensor of energy homeostasis, in a screen for kinases that phosphorylate the Thr 278 site of VASP in endothelial cells. Pharmacological AMPK inhibitors and activators and AMPK mutants revealed that the kinase specifically targets residue Thr-278 but not Ser-157 or Ser-239. Quantitative fluorescence-activated cell sorter analysis and serum response factor transcriptional reporter assays, which quantify the cellular F-/G-actin equilibrium, indicated that AMPK-mediated VASP phosphorylation impaired actin stress fiber formation and altered cell morphology. In the Zucker Diabetic Fatty (ZDF) rat model for type II diabetes, AMPK activity and Thr-278 phosphorylation were substantially reduced in arterial vessel walls. These findings suggest that VASP is a new AMPK substrate, that VASP Thr-278 phosphorylation translates metabolic signals into actin cytoskeleton rearrangements, and that this signaling system becomes down-regulated in diabetic vessels. PMID- 17082197 TI - Alpha-actinin-4 is required for normal podocyte adhesion. AB - Mutations in the alpha-actinin-4 gene ACTN4 cause an autosomal dominant human kidney disease. Mice deficient in alpha-actinin-4 develop a recessive phenotype characterized by kidney failure, proteinuria, glomerulosclerosis, and retraction of glomerular podocyte foot processes. However, the mechanism by which alpha actinin-4 deficiency leads to glomerular disease has not been defined. Here, we examined the effect of alpha-actinin-4 deficiency on the adhesive properties of podocytes in vivo and in a cell culture system. In alpha-actinin-4-deficient mice, we observed a decrease in the number of podocytes per glomerulus compared with wild-type mice as well as the presence of podocyte markers in the urine. Podocyte cell lines generated from alpha-actinin-4-deficient mice were less adherent than wild-type cells to glomerular basement membrane (GBM) components collagen IV and laminin 10 and 11. We also observed markedly reduced adhesion of alpha-actinin-4-deficient podocytes under increasing shear stresses. This adhesion deficit was restored by transfecting cells with alpha-actinin-4-GFP. We tested the strength of the integrin receptor-mediated linkages to the cytoskeleton by applying force to microbeads bound to integrin using magnetic pulling cytometry. Beads bound to alpha-actinin-4-deficient podocytes showed greater displacement in response to an applied force than those bound to wild type cells. Consistent with integrin-dependent alpha-actinin-4-mediated adhesion, phosphorylation of beta1-integrins on alpha-actinin-4-deficient podocytes is reduced. We rescued the phosphorylation deficit by transfecting alpha-actinin-4 into alpha-actinin-4-deficient podocytes. These results suggest that alpha actinin-4 interacts with integrins and strengthens the podocyte-GBM interaction thereby stabilizing glomerular architecture and preventing disease. PMID- 17082198 TI - Right ventromedial prefrontal lesions result in paradoxical cardiovascular activation with emotional stimuli. AB - Ventromedial prefrontal cortex (VMPFC) lesions can alter emotional and autonomic responses. In animals, VMPFC activation results in cardiovascular sympathetic inhibition. In humans, VMPFC modulates emotional processing and autonomic response to arousal (e.g. accompanying decision-making). The specific role of the left or right VMPFC in mediating somatic responses to non-arousing, daily-life pleasant or unpleasant stimuli is unclear. To further evaluate VMPFC interaction with autonomic processing of non-stressful emotional stimuli and assess the effects of stimulus valence, we studied patients with unilateral VMPFC lesions and assessed autonomic modulation at rest and during physical challenge, and heart rate (HR) and blood pressure (BP) responses to non-stressful neutral, pleasant and unpleasant visual stimulation (VES) via emotionally laden slides. In 6 patients (54.0 +/- 7.2 years) with left-sided VMPFC lesions (VMPFC-L), 7 patients (43.3 +/- 11.6 years) with right-sided VMPFC lesions (VMPFC-R) and 13 healthy volunteers (44.7 +/- 11.6 years), we monitored HR as R-R interval (RRI), BP, respiration, end-tidal carbon dioxide levels, and oxygen saturation at rest, during autonomic challenge by metronomic breathing, a Valsalva manoeuvre and active standing, and in response to non-stressful pleasant, unpleasant and neutral VES. Pleasantness versus unpleasantness of slides was rated on a 7-point Likert scale. At rest, during physical autonomic challenge, and during neutral VES, parameters did not differ between the patient groups and volunteers. During VES, Likert scores also were similar across the three groups. During pleasant and unpleasant VES, HR decreased (i.e. RRI increased) significantly whereas BP remained unchanged in volunteers. In VMPFC-L patients, HR decrease was insignificant with pleasant and unpleasant VES. BP slightly increased (P = 0.06) with pleasant VES but was stable with unpleasant VES. In contrast, VMPFC-R patients had significant increases in HR and BP during pleasant and not quite significant HR increases (P = 0.06) with only slight BP increase during unpleasant VES. Other biosignals remained unchanged during VES in all groups. Our results show that VMPFC has no major influence on autonomic modulation at rest and during non-emotional, physical stimulation. The paradoxical HR and BP responses in VMPFC-R patients suggest hemispheric specialization for VMPFC interaction with predominant parasympathetic activation by the left, but sympathetic inhibition by the right VMPFC. Valence of non-stressful stimuli has a limited effect with more prominent left VMPFC modulation of pleasant and more right VMPFC modulation of unpleasant stimuli. The paradoxical sympathetic disinhibition in VMPFC-R patients may increase their risk of sympathetic hyperexcitability with negative consequences such as anxiety, hypertension or cardiac arrhythmias. PMID- 17082199 TI - Assessing seizure dynamics by analysing the correlation structure of multichannel intracranial EEG. AB - Epileptic seizures are commonly characterized as 'hypersynchronous states'. This habit is doubly misleading, because seizures are not necessarily synchronous and are not unchanging 'states' but dynamic processes. Here the temporal evolution of the correlation structure in the course of 100 focal onset seizures of 60 patients recorded by intracranial multichannel EEG was assessed. To this end a multivariate method was applied that at its core consists of computing the eigenvalue spectrum of the zero-lag correlation matrix of a short sliding window. Our results show that there are clearly observable and statistically significant changes of the correlation structure of focal onset seizures. Specifically, these changes indicate that the zero-lag correlation of multi-channel EEG either remains approximately unchanged or-especially in the case of secondary generalization-decreases during the first half of the seizures. Then correlation gradually increases again before the seizures terminate. This development was qualitatively independent of the anatomical location of the seizure onset zone and therefore seems to be a generic property of focal onset seizures. We suggest that the decorrelation of EEG activity is due to the different propagation times of locally synchronous ictal discharges from the seizure onset zone to other brain areas. Furthermore we speculate that the increase of correlation during the second half of the seizures may be causally related to seizure termination. PMID- 17082200 TI - Epilepsy surgery involving the sensory-motor cortex. AB - Our aim was to assess the outcome with regard to seizures and neurological function in unselected patients undergoing resective surgery involving the perirolandic area, with or without multiple subpial transections (MSTs). All patients who underwent perirolandic cortical resection or MSTs from 1979 to 2003 at the London Health Sciences Centre were identified. Patients were included if they had seizures originating in the perirolandic area, recorded with subdural electrodes, or if they had scalp recorded seizures and a congruent discrete epileptogenic lesion on MRI in the perirolandic area. Most patients had electrocorticography (ECoG) at the time of surgery. Data collected include pre operative and post-operative neurological deficits, MRI findings, interictal and ictal scalp EEG, interictal and ictal subdural data, ECoG findings, type and extent of surgery, neuropathologic diagnoses, and seizure outcomes. We studied 52 patients (22 females). The average age at the time of surgery was 33 years, and the average post-operative follow-up was 4.2 years. The most frequent aetiologies were neoplastic in 26 patients (50%), vascular in eight (15%), malformations of cortical development in six (12%), Rasmussen's encephalitis in three (6%) and other aetiologies in nine (17%). Surgery involved the pre-central gyrus in 17 patients, pre- and post-central gyrus in 13, the inferior central region in 11, the post-central gyrus in 7, and the pre-central gyrus and mesial frontal area in 2. At last follow-up 16 patients were in Engel class I (31%), 8 (15%) in class II, 14 (27%) in class III and 14 (27%) in class IV. Residual neurological deficits were present in 26 patients (50%), occurred more frequently in patients > or =25 years old (P < 0.05) and were mild in 14 (54%) patients. In univariate analyses, better seizure outcomes (P < 0.05) occurred in patients whose ECoG showed infrequent post-resectional spikes and no spikes distant to the resection margin, and in resections involving the pre-central and inferior rolandic cortex. In unselected patients with intractable perirolandic epilepsy, many of whom have large, complex epileptogenic lesions, various levels of seizure improvement can be achieved in almost 75% through well-planned surgical resections. New, severe post-operative neurological deficits can occur in 23% of these patients and appear to be more frequent in older patients. Whereas scalp EEG provided limited information to guide surgery, findings on interictal ECoG predicted seizure outcome. PMID- 17082201 TI - Acinetobacter baumannii bloodstream infection while receiving tigecycline: a cautionary report. AB - OBJECTIVES: Tigecycline has shown in vitro activity against Acinetobacter baumannii. Yet, published clinical experience with tigecycline use outside clinical trials is lacking. We describe, for the first time, bloodstream infection caused by tigecycline-non-susceptible A. baumannii occurring in patients receiving tigecycline for other indications. The possible mechanisms of resistance and pharmacokinetic limitations of the drug are addressed. METHODS: The clinical records of involved patients were systematically reviewed. Tigecycline susceptibility testing was initially performed using the Etest method and confirmed by agar dilution. Involved isolates underwent PFGE and exposure to phenyl-arginine-beta-naphthylamide (PAbetaN), an efflux pump inhibitor. RESULTS: Two patients developed A. baumannii bloodstream infection while receiving tigecycline. Tigecycline was administered for other indications for 9 and 16 days, respectively, before the onset of A. baumannii infection. Patient 1 died of overwhelming A. baumannii infection and Patient 2 recovered after a change in antibiotic therapy. The MICs of tigecycline were 4 and 16 mg/L, respectively. Both isolates had a multidrug-resistant phenotype and were genotypically unrelated. After exposure to PAbetaN, the MICs reduced to 1 and 4 mg/L, respectively. CONCLUSIONS: To our knowledge, this is the first clinical description of bloodstream infection caused by tigecycline-non-susceptible A. baumannii. Such resistance appears to be at least partly attributable to an efflux pump mechanism. Given the reported low serum tigecycline levels, we urge caution when using this drug for treatment of A. baumannii bloodstream infection. PMID- 17082202 TI - The potential role of computerized decision support systems to improve empirical antibiotic prescribing. AB - Prudent antibiotic prescribing is a cornerstone of efforts to control the emergence and spread of antimicrobial resistance and to reduce the occurrence of Clostridium difficile associated diarrhoea, and is also currently the subject of a joint Working Party of the British Society for Antimicrobial Chemotherapy and the Hospital Infection Society. An article in this issue of the Journal describes a cluster randomized trial of an intervention to improve antibiotic prescribing using TREAT, a computerized decision support system. The results from this trial provide a significant contribution to the development of evidence-based prescribing of antibiotics. PMID- 17082203 TI - U12DB: a database of orthologous U12-type spliceosomal introns. AB - U12-type introns are spliced by the U12-dependent spliceosome and are present in the genomes of many higher eukaryotic lineages including plants, chordates and some invertebrates. However, due to their relatively recent discovery and a systematic bias against recognition of non-canonical splice sites in general, the introns defined by U12-type splice sites are under-represented in genome annotations. Such under-representation compounds the already difficult problem of determining gene structures. It also impedes attempts to study these introns genome-wide or phylum-wide. The resource described here, the U12 Intron Database (U12DB), aims to catalog the U12-type introns of completely sequenced eukaryotic genomes in a framework that groups orthologous introns with each other. This will aid further investigations into the evolution and mechanism of U12-dependent splicing as well as assist ongoing genome annotation efforts. Public access to the U12DB is available at http://genome.imim.es/cgi-bin/u12db/u12db.cgi. PMID- 17082204 TI - NATsDB: Natural Antisense Transcripts DataBase. AB - Natural antisense transcripts (NATs) are reverse complementary at least in part to the sequences of other endogenous sense transcripts. Most NATs are transcribed from opposite strands of their sense partners. They regulate sense genes at multiple levels and are implicated in various diseases. Using an improved whole genome computational pipeline, we identified abundant cis-encoded exon overlapping sense-antisense (SA) gene pairs in human (7356), mouse (6806), fly (1554), and eight other eukaryotic species (total 6534). We developed NATsDB (Natural Antisense Transcripts DataBase, http://natsdb.cbi.pku.edu.cn/) to enable efficient browsing, searching and downloading of this currently most comprehensive collection of SA genes, grouped into six classes based on their overlapping patterns. NATsDB also includes non-exon-overlapping bidirectional (NOB) genes and non-bidirectional (NBD) genes. To facilitate the study of functions, regulations and possible pathological implications, NATsDB includes extensive information about gene structures, poly(A) signals and tails, phastCons conservation, homologues in other species, repeat elements, expressed sequence tag (EST) expression profiles and OMIM disease association. NATsDB supports interactive graphical display of the alignment of all supporting EST and mRNA transcripts of the SA and NOB genes to the genomic loci. It supports advanced search by species, gene name, sequence accession number, chromosome location, coding potential, OMIM association and sequence similarity. PMID- 17082205 TI - InSatDb: a microsatellite database of fully sequenced insect genomes. AB - InSatDb presents an interactive interface to query information regarding microsatellite characteristics per se of five fully sequenced insect genomes (fruit-fly, honeybee, malarial mosquito, red-flour beetle and silkworm). InSatDb allows users to obtain microsatellites annotated with size (in base pairs and repeat units); genomic location (exon, intron, up-stream or transposon); nature (perfect or imperfect); and sequence composition (repeat motif and GC%). One can access microsatellite cluster (compound repeats) information and a list of microsatellites with conserved flanking sequences (microsatellite family or paralogs). InSatDb is complete with the insects information, web links to find details, methodology and a tutorial. A separate 'Analysis' section illustrates the comparative genomic analysis that can be carried out using the output. InSatDb is available at www.cdfd.org.in/insatdb. PMID- 17082206 TI - MACiE (Mechanism, Annotation and Classification in Enzymes): novel tools for searching catalytic mechanisms. AB - MACiE (Mechanism, Annotation and Classification in Enzymes) is a database of enzyme reaction mechanisms, and is publicly available as a web-based data resource. This paper presents the first release of a web-based search tool to explore enzyme reaction mechanisms in MACiE. We also present Version 2 of MACiE, which doubles the dataset available (from Version 1). MACiE can be accessed from http://www.ebi.ac.uk/thornton-srv/databases/MACiE/ PMID- 17082207 TI - Predicting survival with good neurological recovery at hospital admission after successful resuscitation of out-of-hospital cardiac arrest: the OHCA score. AB - AIMS: Out-of-hospital cardiac arrest (OHCA) is common and carries a bleak prognosis. Early prediction of unfavourable outcomes is difficult but crucial to improve resource allocation. The aim of this study was to develop a simple tool for predicting survival with good neurological function in the overall population of patients with successfully resuscitated cardiac arrest. METHODS AND RESULTS: We used logistic regression analysis to identify clinical and laboratory variables that were both readily available at admission and predictive of poor outcomes (death or severe neurological impairment) in a development cohort of 130 consecutive OHCA patients admitted to a French intensive care unit (ICU) between 1999 and 2003. To test the prediction score built from these variables, we used a validation cohort of 210 patients recruited in four French ICUs between 2003 and 2005. Initial rhythm, estimated no-flow and low-flow intervals, blood lactate, and creatinine levels determined using whole blood analyzers were independently associated with poor outcomes and were used to build a continuous severity score. Goodness-of-fit tests indicated good performance (P=0.79 in the development cohort and P=0.13 in the validation cohort). The area under the receiver operating characteristics curve was 0.82 in the development cohort and 0.88 in the validation cohort. CONCLUSION: The outcome can be accurately predicted after OHCA using variables that are readily available at ICU admission. PMID- 17082208 TI - Depression as an aetiologic and prognostic factor in coronary heart disease: a meta-analysis of 6362 events among 146 538 participants in 54 observational studies. AB - AIMS: With negative treatment trials, the role of depression as an aetiological or prognostic factor in coronary heart disease (CHD) remains controversial. We quantified the effect of depression on CHD, assessing the extent of confounding by coronary risk factors and disease severity. METHODS AND RESULTS: Meta-analysis of cohort studies measuring depression with follow-up for fatal CHD/incident myocardial infarction (aetiological) or all-cause mortality/fatal CHD (prognostic). We searched MEDLINE and Science Citation Index until December 2003. In 21 aetiological studies, the pooled relative risk of future CHD associated with depression was 1.81 (95% CI 1.53-2.15). Adjusted results were included for 11 studies, with adjustment reducing the crude effect marginally from 2.08 (1.69 2.55) to 1.90 (1.49-2.42). In 34 prognostic studies, the pooled relative risk was 1.80 (1.50-2.15). Results adjusted for left ventricular function result were available in only eight studies; and this attenuated the relative risk from 2.18 to 1.53 (1.11-2.10), a 48% reduction. Both aetiological and prognostic studies without adjusted results had lower unadjusted effect sizes than studies from which adjusted results were included (P<0.01). CONCLUSION: Depression has yet to be established as an independent risk factor for CHD because of incomplete and biased availability of adjustment for conventional risk factors and severity of coronary disease. PMID- 17082209 TI - Intracoronary thrombus in a 26-year-old man. PMID- 17082210 TI - Using genomics and proteomics to investigate mechanisms of transcriptional silencing in Saccharomyces cerevisiae. AB - Silent chromatin in budding yeast is characterized by the presence of a specialized chromatin modification complex consisting of silent information regulator (Sir) proteins, closely packed pairs of nucleosomes, and hypoacetylated and hypomethylated histones. How this specialized chromatin is established, maintained and inherited has been extensively studied. Less investigated are the determinants that constrain its linear spread along the chromatin fibre and the manner by which it represses gene transcription. Here we review the essential features of SIR-mediated heterochromatin, and discuss genomic and proteomic approaches for discerning the composition of its boundaries and for elucidating the mechanisms by which it silences transcription. PMID- 17082211 TI - Uraemic plasma decreases the expression of ABCA1, ABCG1 and cell-cycle genes in human coronary arterial endothelial cells. AB - BACKGROUND: Uraemia is associated with endothelial dysfunction, but the effect of uraemic plasma on the gene expression pattern of human coronary arterial endothelial cells (HCAEC) has never been defined. METHODS: HCAECs were exposed for 48 h to a culture medium supplemented with 20% uraemic vs normal plasma. We extracted mRNA and hybridized it onto Affymetrix HG-U133 Plus2 microarrays. We validated our findings for five genes of interest by real-time PCR and performed evaluations of cell proliferation and apoptosis in HCAECs exposed to uraemic vs normal plasma. RESULTS: Six genes involved in the regulation of cell-cycle progression (CDK-1, topoisomerase II, PDZ-binding kinase, CDCA1, protein SDP35, E2F transcription factor 8) and two genes of the cholesterol efflux system (ABCA1 and ABCG1) were down-regulated in HCAECs exposed to uraemic plasma (>1.75-fold change vs normal). Real-time PCR confirmed the down-regulation observed in the microarray experiment. Cell proliferation was significantly decreased in HCAECs exposed to uraemic vs normal plasma for 48 h (86 vs 95% of serum-starved control, P = 0.006). Exposure to uraemic plasma for 48 h was associated with increased apoptosis of HCAEC as compared with normal plasma (7.7 vs 2.8%, P < 0.001), a phenomenon that was further enhanced when oxidized LDLs (150 microg protein/ml) were added to the medium containing uraemic plasma (16.9 vs 7.7%, P < 0.001). CONCLUSIONS: The down-regulation of genes involved in cell-cycle progression and cholesterol efflux from HCAECs exposed to uraemic conditions could contribute to enhancing endothelial dysfunction and atherosclerosis in patients with chronic renal failure. PMID- 17082212 TI - Cardiac tamponade--knowing how far to insert the wire. PMID- 17082213 TI - Subclinical hypothyroidism is linked to micro-inflammation and predicts death in continuous ambulatory peritoneal dialysis. AB - BACKGROUND: Low T3 is a frequent alteration in patients with ESRD. This derangement has been recently linked to inflammation in haemodialysis patients. Whether this association holds true in peritoneal dialysis patients has not been studied. METHODS: We investigated the relationship between low-grade inflammation [IL-6, C-reactive protein (CRP) and serum albumin levels] and free tri iodothyronine (fT3) in a cohort of 41 CAPD patients (mean age, 66 years; M, 26; F, 15) without heart failure and inter-current illnesses. RESULTS: CAPD patients had lower fT3 levels (2.7 +/- 0.8 pg/ml) than healthy subjects (3.7 +/- 1.0 pg/ml, P < 0.001) of similar age. Free T3 levels were directly related to those of serum albumin (r = 0.52, P = 0.001) and inversely to IL-6 (r = -0.30, P = 0.05) and CRP (r = -0.54, P < 0.001). Age (r = -0.61, P < 0.001), haemoglobin levels (r = 0.32, P = 0.05) and diastolic blood pressure (r = 0.50, P = 0.001) were also related to fT3. In multiple regression models adjusting for all variables related to fT3, CRP and albumin were retained as independent correlates of fT3. During the follow-up (2.8 +/- 1.7 years) 27 patients died. Plasma fT3 levels were lower in patients who died (2.5 +/- 0.8 pg/ml) compared with survivors (3.3 +/- 0.5 pg/ml P = 0.001). In Cox analyses, fT3 was a significant predictor of mortality independent of the main traditional as well as non traditional risk factors. CONCLUSIONS: The relationship between fT3, CRP and serum albumin suggests that inflammation-malnutrition might be involved in the low T3 syndrome in CAPD patients. Thyroid dysfunction might be implicated in the pathogenic pathway which links micro-inflammation to survival in PD patients. PMID- 17082214 TI - Primary biliary cirrhosis associated with minimal change disease. PMID- 17082215 TI - Expression and Ca2+ dependency of plasma membrane K+ channels of tobacco suspension cells adapted to salt stress. AB - The expression of plasma membrane K+ channels of NaCl-adapted tobacco suspension cells and effects of extracellular Ca2+ on plasma membrane K+ channels were investigated. Reverse transcription-PCR (RT-PCR) analysis showed that expression of TORK1, which encodes a K+ channel, was much lower in NaCl-adapted cells than in NaCl-unadapted cells. The magnitude of the outward K+ currents of NaCl-adapted as well as NaCl-unadapted cells decreased with increasing extracellular Ca2+ but there is no significant difference in Ca2+ dependency of the K+ current. These analyses suggest that reduction of the number of K+ channels might cause NaCl adaptation of cells through the decrease of outward K+ currents. PMID- 17082216 TI - Chloroplast NAD kinase is essential for energy transduction through the xanthophyll cycle in photosynthesis. AB - Photosynthetic parameters of the nadk2 mutant of Arabidopsis thaliana, which is defective in chloroplast NAD kinase, were investigated. In this plant, the effective efficiency of photosynthetic electron transport (PhiII) and the quantum yield of open reaction centers of photosystem II (Fv'/Fm') were decreased. Furthermore, an increase in non-photochemical quenching attributed to energy dissipation from the xanthophyll cycle was observed. The mutant showed an aberrant de-epoxidation state of xanthophyll cycle carotenoids and had a high level of zeaxanthin even under low light conditions. These results indicate that chloroplast NAD kinase, catalyzing phosphorylation of NAD, is essential for the proper photosynthetic machinery of PSII and the xanthophyll cycle. PMID- 17082217 TI - Did J. Marion Sims deliberately addict his first fistula patients to opium? AB - American surgeon J. Marion Sims (1813-83) is regarded by many modern authors as a controversial figure because he carried out a series of experimental surgeries on enslaved African American women between 1846 and 1849 in an attempt to cure them of vesicovaginal fistulas, which they had all developed as a result of prolonged obstructed labor. He operated on one woman, Anarcha Westcott, thirty times before he successfully closed her fistula. Sims performed these fistula repair operations without benefit of anesthesia but gave these women substantial doses of opium afterwards. Several modern writers have alleged that Sims did this in order to addict them to the drug and thereby to enhance his control over them. This article examines the controversy surrounding Sims' use of postoperative opium in these enslaved surgical patients. The evidence suggests that although these women were probably tolerant to the doses of opium that he used, there is no evidence that he deliberately tried to addict them to this drug. Sims' use of postoperative opium appears to have been well supported by the therapeutic practices of his day, and the regimen that he used was enthusiastically supported by many contemporary surgeons. PMID- 17082218 TI - Treatment outcome of metastatic testicular cancer at a single institution in Japan, a country with low incidence of germ cell tumor. AB - BACKGROUND: Incidence of testicular cancer is low in Japan. Recent investigations showed that the outcome of metastatic testicular cancer is associated with the experience of the treatment institution. Objectives of the present investigation are to evaluate outcome of a single institution in Japan, and to identify prognostic factors for testicular cancer. METHODS: We retrospectively analysed the outcome of 74 patients with metastatic testicular cancer who were treated at Tsukuba University Hospital (TUH) between January 1981 and January 2003. Forty five patients (61%) were referred to the TUH for the treatment of metastatic disease. The progression-free survival (PFS) rates according to the International Germ Cell Cancer (IGCC) classification and the Indiana University classification were used to evaluate the treatment outcome. RESULTS: The median follow-up period of all patients was 87 months (range, 13-260 months). Forty patients (54%) were classified as having good prognosis, 20 (27%) intermediate, and 14 (19%) poor. The overall 5- and 10-year PFS was 79 and 74%, respectively. The 5-year PFS with good, intermediate and poor prognosis was 90, 70 and 64%, respectively. There was a significant difference between the three groups (P=0.02), but the survival of the intermediate-prognosis group was not statistically different from that of the poor-prognosis group. The Indiana University classification failed to discriminate the prognoses of moderate and advanced disease, but proved to be an independent prognostic factor for progression-free survival in intermediate- and poor-prognosis patients (P=0.025, hazard ratio=5.39). CONCLUSIONS: Our treatment outcome is not different from that of the institutions participating in the International Germ Cell Cancer Consensus Group (IGCCCG). PMID- 17082219 TI - A randomized trial comparing radical prostatectomy plus endocrine therapy versus external beam radiotherapy plus endocrine therapy for locally advanced prostate cancer: results at median follow-up of 102 months. AB - BACKGROUND: To investigate the optimal treatment of locally advanced prostate cancer, a prospective randomized trial was conducted to compare radical prostatectomy plus endocrine therapy versus external beam radiotherapy plus endocrine therapy. METHODS: One hundred patients with T2b-3N0M0 prostate cancer were enrolled and 95 were evaluated. Of 95 cases, 46 underwent radical prostatectomy with pelvic lymph node dissection and 49 were treated with external beam radiation by linear accelerator with 40-50 Gy to the whole pelvis and 20-Gy boost to the prostatic area. For all patients, endocrine therapy was initiated 8 weeks before surgery or radiotherapy and continued thereafter. The long-term outcome and morbidity were examined. RESULTS: Median follow-up period was 102 months. At 10 years overall survival rates in the surgery group were better than the radiation group (76.2% versus 71.1% for biochemical progression-free rates; P=0.25, 83.5% versus 66.1% for clinical progression-free rates; P=0.14, 85.7% versus 77.1% for cause-specific survival rates; P=0.06, and 67.9% versus 60.9% for overall survival rates; P=0.30), although none of them reached statistical significance. Erectile dysfunction was recognized in almost all patients as a result of continuous endocrine therapy. Incontinence requiring more than one pad per day was observed more frequently in the surgery group than the radiation group (P<0.01). CONCLUSIONS: For the treatment of patients with locally advanced prostate cancer, when combined with endocrine therapy, either radical prostatectomy or external beam radiotherapy demonstrated favorable long-term outcomes. The radiation dose of 60-70 Gy might not be enough for the local treatment of locally advanced prostate cancer. PMID- 17082220 TI - Isolation and expression profiling of genes upregulated in bone marrow-derived mononuclear cells of rheumatoid arthritis patients. AB - We have comprehensively identified the genes whose expressions are augmented in bone marrow-derived mononuclear cells (BMMC) from patients with Rheumatoid Arthritis (RA) as compared with BMMCs from Osteoarthritis (OA) patients, and named them AURA after augmented in RA. Both stepwise subtractive hybridization and microarray analyses were used to identify AURA genes, which were confirmed by northern blot analysis and/or reverse transcription polymerase chain reaction (RT PCR). We also assessed their expression levels in individual patients by quantitative real-time RT-PCR. Of 103 AURA genes we have identified, the mRNA levels of the following 10 genes, which are somehow related to immune responses, were increased in many of the RA patients: AREG (=AURA9), FK506-binding protein 5 (FKBP5 = AURA45), C-type lectin superfamily member 9 (CLECSF9 = AURA24), tyrosylprotein sulfotransferase 1 (TPST1 = AURA52), lymphocyte G0/G1 switch gene (G0S2 = AURA8), chemokine receptor 4 (CXCR4 = AURA86), nuclear factor-kappa B (NF kappaB = AURA25) and two genes of unknown function (FLJ11106 = AURA1, BC022398 = AURA2 and XM_058513 = AURA17). Since AREG was most significantly increased in many of the RA patients, we subjected it to further analysis and found that AREG epidermal growth factor receptor signaling is highly activated in synovial cells isolated from RA patients, but not in OA synoviocytes. We propose that the expression profiling of these AURA genes may improve our understanding of the pathogenesis of RA. PMID- 17082221 TI - The efficiency of magnetic-activated cell sorting and fluorescence-activated cell sorting in the decontamination of testicular cell suspensions in cancer patients. AB - BACKGROUND: Before clinical application, the feasibility and safety of autologous testicular stem cell transplantation should be explored. Apart from limitations in their numbers, spermatogonial stem cells may also be contaminated by malignant cells. Therefore, both enrichment and decontamination before transplantation may be necessary. This study aimed at evaluating the decontaminating potential of magnetic-activated cell sorting (MACS) and/or fluorescence-activated cell sorting (FACS) for both murine and human testicular cell suspensions. In the mouse, the effectiveness of the transplantation technique after cell sorting was also assessed. METHODS: Murine testicular cells were contaminated with 5% EL4 cells. Fresh and frozen-thawed suspensions were sorted using MACS (CD49f (+)) and FACS (CD49f(+), H-2Kb(-)) and evaluated by FACS, cell culture and transplantation into W/W(v) mice. Human testicular cells were contaminated with 5 or 0.05% CCRF-SB (SB) cells. Frozen-thawed suspensions were sorted using FACS (HLA class I(-)) and evaluated by FACS, cell culture and PCR for the B-cell receptor. RESULTS: In the mouse, the sorted fractions contained 0.39% H-2K(b)-positive and 76.55% CD49f positive cells. After transplantation, 1 in 20 recipient mice developed a malignancy. In the human experiments, an average of 0.58% SB cells was detected after sorting. In only 1 of 11 samples, there were no SB cells observed. CONCLUSION: MACS and/or FACS are insufficient for completely depleting testicular tissue of malignant cells. Although more research on alternative decontamination techniques is necessary, developing a reliable method to screen a priori testicular tissue for malignant cells may be equally important. PMID- 17082222 TI - Dietary nitrate increases gastric mucosal blood flow and mucosal defense. AB - Salivary nitrate from dietary or endogenous sources is reduced to nitrite by oral bacteria. In the acidic stomach, nitrite is further reduced to bioactive nitrogen oxides, including nitric oxide (NO). In this study, we investigated the gastroprotective role of nitrate intake and of luminally applied nitrite against provocation with diclofenac and taurocholate. Mucosal permeability ((51)Cr-EDTA clearance) and gastric mucosal blood flow (laser-Doppler flowmetry) were measured in anesthetized rats, either pretreated with nitrate in the drinking water or given acidified nitrite luminally. Diclofenac was given intravenously and taurocholate luminally to challenge the gastric mucosa. Luminal NO content and nitrite content in the gastric mucus were determined by chemiluminescence. The effect of luminal administration of acidified nitrite on the mucosal blood flow was also investigated in endothelial nitric oxide synthase-deficient mice. Rats pretreated with nitrate or given nitrite luminally had higher gastric mucosal blood flow than controls. Permeability increased more during the provocation in the controls than in the nitrate- and nitrite-treated animals. Dietary nitrate increased luminal NO levels 50 times compared with controls. Nitrate intake also resulted in nitrite accumulation in the loosely adherent mucous layer; after removal of this mucous layer, blood flow was reduced. Nitrite administrated luminally in endothelial nitric oxide synthase-deficient mice increased mucosal blood flow. We conclude that dietary nitrate and direct luminal application of acidified nitrite decrease diclofenac- and taurocholate-induced mucosal damage. The gastroprotective effect likely involves a higher mucosal blood flow caused by nonenzymatic NO production. These data suggest an important physiological role of nitrate in the diet. PMID- 17082223 TI - Two N-linked glycans are required to maintain the transport activity of the bile salt export pump (ABCB11) in MDCK II cells. AB - The aim of this study was to determine the role of N-linked glycosylation in protein stability, intracellular trafficking, and bile acid transport activity of the bile salt export pump [Bsep (ATP-binding cassette B11)]. Rat Bsep was fused with yellow fluorescent protein, and the following mutants, in which Asn residues of putative glycosylation sites (Asn(109), Asn(116), Asn(122), and Asn(125)) were sequentially replaced with Gln, were constructed by site-directed mutagenesis: single N109Q, double N109Q + N116Q, triple N109Q + N116Q + N122Q, and quadruple N109Q + N116Q + N122Q + N125Q. Immunoblot and glycosidase cleavage analysis demonstrated that each site was glycosylated. Removal of glycans decreased taurocholate transport activity as determined in polarized MDCK II cells. This decrease resulted from rapid decay of the mutant Bsep protein; biochemical half lives were 3.76, 3.65, 3.24, 1.35, and 0.52 h in wild-type, single-mutant, double mutant, triple-mutant, and quadruple-mutant cells, respectively. Wild-type and single- and double-mutant proteins were distributed exclusively along the apical membranes, whereas triple- and quadruple-mutant proteins remained intracellular. MG-132 but not bafilomycin A(1) extended the half-life, suggesting a role for the proteasome in Bsep degradation. To determine whether a specific glycosylation site or the number of glycans was critical for protein stability, we studied the protein expression of combinations of N-glycan-deficient mutants and observed that Bsep with one glycan was considerably unstable compared with Bsep harboring two or more glycans. In conclusion, at least two N-linked glycans are required for Bsep protein stability, intracellular trafficking, and function in the apical membrane. PMID- 17082224 TI - TLRS in the gut. II. Flagellin-induced inflammation and antiapoptosis. AB - Flagellin is bacterial protein that serves as a danger signal across a wide variety of eukaryotes and is a potent inducer of inflammatory effector responses in the mammalian gut. Recent findings utilizing purified flagellin and flagellate/aflagellate bacteria in in vitro and in vivo systems have revealed the important roles played by flagellin in the initial encounter between mucosa and flagellate bacteria, specifically in the modulation of apoptotic responses. PMID- 17082225 TI - Concise review: therapeutic strategies for Parkinson disease based on the modulation of adult neurogenesis. AB - Parkinson disease (PD) is a progressive neurodegenerative disorder affecting millions of people worldwide. To date, treatment strategies are mainly symptomatic and aimed at increasing dopamine levels in the degenerating nigrostriatal system. Hope rests upon the development of effective neurorestorative or neuroregenerative therapies based on gene and stem cell therapy or a combination of both. The results of experimental therapies based on transplanting exogenous dopamine-rich fetal cells or glial cell line-derived neurotrophic factor overexpression into the brain of Parkinson disease patients encourage future cell- and gene-based strategies. The endogenous neural stem cells of the adult brain provide an alternative and attractive cell source for neuroregeneration. Prior to designing endogenous stem cell therapies, the possible impact of PD on adult neuronal stem cell pools and their neurogenic potential must be investigated. We review the experimental data obtained in animal models or based on analysis of patients' brains prior to describing different treatment strategies. Strategies aimed at enhancing neuronal stem cell proliferation and/or differentiation in the striatum or the substantia nigra will have to be compared in animal models and selected prior to clinical studies. PMID- 17082226 TI - RE1 Silencing transcription factor maintains a repressive chromatin environment in embryonic hippocampal neural stem cells. AB - The control of gene expression in neural stem cells is key to understanding their developmental and therapeutic potential, yet we know little of the transcriptional mechanisms that underlie their differentiation. Recent evidence has implicated the RE1 silencing transcription factor (REST) in neuronal differentiation. However, the means by which REST regulates transcription in neural stem cells remain unclear. Here, we show that REST recruits distinct corepressor platforms in neural stem cells. REST is able to both silence and repress neuronal genes in embryonic hippocampal neural stem cells by creating a chromatin environment that contains both repressive local epigenetic signature (characterized by low levels of histones H4 and H3K9 acetylation and elevated dimethylation of H3K9) and H3K4 methylation, which are characteristic of gene activation. Furthermore, inhibition of REST function leads to activation of several neuron-specific genes but does not lead to overt formation of mature neurons, supporting the notion that REST regulates part, but not all, of the neuronal differentiation program. PMID- 17082227 TI - Homosynaptic and heterosynaptic modes of endocannabinoid action at hippocampal CCK+ basket cell synapses. PMID- 17082228 TI - Cellular mechanisms of burst firing-mediated long-term depression in rat neocortical pyramidal cells. AB - During wakefulness and sleep, neurons in the neocortex emit action potentials tonically or in rhythmic bursts, respectively. However, the role of synchronized discharge patterns is largely unknown. We have recently shown that pairings of excitatory postsynaptic potentials (EPSPs) and action potential bursts or single spikes lead to long-term depression (burst-LTD) or long-term potentiation, respectively. In this study, we elucidate the cellular mechanisms of burst-LTD and characterize its functional properties. Whole-cell patch-clamp recordings were obtained from layer V pyramidal cells in somatosensory cortex of juvenile rats in vitro and composite EPSPs and EPSCs were evoked extracellularly in layers II/III. Repetitive burst-pairings led to a long-lasting depression of EPSPs and EPSCs that was blocked by inhibitors of metabotropic glutamate group 1 receptors, phospholipase C, protein kinase C (PKC) and calcium release from the endoplasmic reticulum, and that required an intact machinery for endocytosis. Thus, burst-LTD is induced via a Ca2+- and phosphatidylinositol-dependent activation of PKC and expressed through phosphorylation-triggered endocytosis of AMPA receptors. Functionally, burst-LTD is inversely related to EPSP size and bursts dominate single spikes in determining the sign of synaptic plasticity. Thus burst-firing constitutes a signal by which coincident synaptic inputs are proportionally downsized. Overall, our data thus suggest a mechanism by which synaptic weights can be reconfigured during non-rapid eye movement sleep. PMID- 17082229 TI - Identifying cardiorespiratory neurocircuitry involved in central command during exercise in humans. AB - For almost one hundred years, the exact role of human brain structures controlling the cardiorespiratory response to exercise ('central command') has been sought. Animal experiments and functional imaging studies have provided clues, but the underlying electrophysiological activity of proposed relevant neural sites in humans has never been measured. In this study, local field potentials were directly recorded in a number of 'deep' brain nuclei during an exercise task designed to dissociate the exercise from peripheral feedback mechanisms. Several patient groups had electrodes implanted sterotaxically for the treatment of movement disorder or chronic pain. Fast Fourier transform analysis was applied to the neurograms to identify the power of fundamental spectral frequencies. Anticipation of exercise resulted in increases in heart rate, blood pressure and ventilation. The greatest neural changes were found in the periaqueductal grey area (PAG) where anticipation of exercise was accompanied by an increase of 43% in the power of the 12-25 Hz frequency band (P = 0.007). Exercise increased the activity by 87% compared to rest (P = 0.006). Changes were also seen in the 60-90 Hz band when anticipation or exercise increased power by 32% (P = 0.006) and 109% (P < 0.001), respectively. In the subthalamic nucleus there was a reduction in the power of the beta frequency during both anticipation (7.6 +/- 0.68% P = 0.001) and exercise (17.3 +/- 0.96% P < 0.001), whereas an increase was seen with exercise only at higher frequencies (93 +/- 1.8% P = 0.007). No significant changes were seen in the globus pallidus during anticipation of exercise. We provide direct electrophysiological evidence highlighting the PAG as an important subcortical area in the neural circuitry of the cardiorespiratory response to exercise, since stimulation of this structure is known to alter blood pressure in awake humans. PMID- 17082230 TI - Unitary IPSPs enhance hilar mossy cell gain in the rat hippocampus. AB - Mechanisms that control neuronal gain allow for adaptive rescaling to synaptic inputs of varying strengths or frequencies. Here, we show that unitary IPSPs (uIPSPs) modulate gain and unitary EPSP (uEPSP)-action potential coupling in mossy cells (MCs) from rat hippocampal slices. Mossy fibre-evoked uEPSCs were large, facilitated and were suppressed by the group II metabotropic glutamate agonist LY354740. Conversely, uIPSCs were smaller, depressed and were not affected by LY354740, but exerted strong inhibitory control over uEPSP-action potential coupling. The IPSC reversal potential was determined by gramicidin perforated patch recordings to be -65.3 +/- 5.0 mV, lying between the resting membrane potential (-75.3 +/- 1.1 mV) and the action potential threshold (-56.5 +/- 2.4 mV). When applied at theta frequency (10 Hz), uIPSPs increased the offset of the MC input-output response to depolarizing current injection, but also increased gain, maximal firing rate and the slope of the depolarization preceding action potentials. These effects were unchanged by the Ca2+ and HCN channel blockers mibefradil and ZD7288, respectively. The height and maximal slope of MC action potentials during tonic depolarization were also increased by uIPSPs, and the decay of uIPSP conductances injected by dynamic clamp at subthreshold membrane potentials was prolonged by TTX. Application of the muscarinic agonist pilocarpine mimicked the effect of IPSPs on MC maximal firing rate, and action potential height and slope, and this was reversed by the GABA(A) antagonist gabazine. Thus, uIPSPs can increase neuronal gain under hyperexcitable conditions, and this effect is probably due to the de-inactivation of a TTX sensitive voltage-dependent Na+ conductance. PMID- 17082231 TI - Ageing and exercise training alter adrenergic vasomotor responses of rat skeletal muscle arterioles. AB - Ageing is associated with increased leg vascular resistance and reductions in leg blood flow during rest and exercise, potentially predisposing older adults to a host of functional and cardiovascular complications. The purpose of these studies was to examine the effects and possible mechanisms of ageing and exercise training on arteriolar adrenergic vasoreactivity. Young and old male Fischer 344 rats were divided into young sedentary (YS), old sedentary (OS), young exercise trained (YT) or old exercise-trained (OT) groups, where training consisted of chronic treadmill exercise. Isolated soleus (SOL) and gastrocnemius (GAS) muscle arterioles were studied in vitro. Responses to noradrenaline in endothelium intact and endothelium-denuded arterioles, as well as during nitric oxide synthase (NOS) inhibition were determined. Vasodilator responses to isoproterenol and forskolin were also determined. RESULTS: Noradrenaline-mediated vasoconstriction was increased in SOL arterioles with ageing, and exercise training in old rats attenuated alpha-adrenergic vasoconstriction in arterioles from both muscle types. Removal of the endothelium and NOS inhibition eliminated these ageing and training effects. Isoproterenol-mediated vasodilatation was impaired with ageing in SOL and GAS arterioles, and exercise training had little effect on this response. Forskolin-induced vasodilatation was not affected by age. The data demonstrate that ageing augments alpha-adrenergic vasoconstriction while exercise training attenuates this response, and both of these alterations are mediated through an endothelial alpha-receptor-NOS-signalling pathway. In contrast, ageing diminishes beta-receptor-mediated vasodilatation, but this impairment is specific to the smooth muscle. These studies indicate that alpha- and beta-adrenergic mechanisms may serve to increase systemic vascular resistance with ageing, and that the effects of exercise training on adrenergic vasomotor properties could contribute to the beneficial effects of exercise on cardiovascular disease. PMID- 17082232 TI - Brainstem PCO2 modulates phrenic responses to specific carotid body hypoxia in an in situ dual perfused rat preparation. AB - Inputs from central (brainstem) and peripheral (carotid body) respiratory chemoreceptors are coordinated to protect blood gases against potentially deleterious fluctuations. However, the mathematics of the steady-state interaction between chemoreceptors has been difficult to ascertain. Further, how this interaction affects time-dependent phenomena (in which chemoresponses depend upon previous experience) is largely unknown. To determine how central P(CO2) modulates the response to peripheral chemostimulation in the rat, we utilized an in situ arterially perfused, vagotomized, decerebrate preparation, in which central and peripheral chemoreceptors were perfused separately (i.e. dual perfused preparation (DPP)). We carried out two sets of experiments: in Experiment 1, we alternated steady-state brainstem P(CO2) between 25 and 50 Torr in each preparation, and applied specific carotid body hypoxia (60 Torr P(O2) and 40 Torr P(CO2)) under both conditions; in Experiment 2, we applied four 5 min bouts (separated by 5 min) of specific carotid body hypoxia (60 Torr P(O2) and 40 Torr P(CO2)) while holding the brainstem at either 30 Torr or 50 Torr P(CO2). We demonstrate that the level of brainstem P(CO2) modulates (a) the magnitude of the phrenic responses to a single step of specific carotid body hypoxia and (b) the magnitude of time-dependent phenomena. We report that the interaction between chemoreceptors is negative (i.e. hypo-additive), whereby a lower brainstem P(CO2) augments phrenic responses resulting from specific carotid body hypoxia. A negative interaction may underlie the pathophysiology of central sleep apnoea in populations that are chronically hypocapnic. PMID- 17082233 TI - Role of vasopressin in rat distal colon function. AB - The specific role of vasopressin in colonic crypt function and its possible synergistic action with aldosterone were studied. Sprague-Dawley rats fed a high Na+ (HS; 150 mM NaCl) or a low-Na+ (LS; 150 microM NaCl) diet were deprived of water or infused with vasopressin, and some animals were treated with specific vasopressin receptor subtype V1 and V2 antagonists. The expression of the epithelial Na+ channel (ENaC), alpha-smooth muscle actin (alpha-SMA) and aquaporin-2 (AQP-2) were determined by immunolocalization in distal colonic mucosa. The pericryptal Na+ concentration was determined by confocal microscopy, using a low-affinity Na+-sensitive fluorescent dye (sodium red) and crypt permeability was measured by the rate of escape of fluorescein isothiocyanate labelled dextran (10 kDa) from the crypt lumen into the pericryptal space in isolated rat distal colonic mucosa. A high plasma concentration of vasopressin raised alpha-SMA expression in the pericryptal sheath (P < 0.05), increased the pericryptal Na+ accumulation in this space (P < 0.01) and caused a reduction of crypt wall permeability (P < 0.01). All these effects were reversed by selective blockade of V1 and V2 receptors. No synergistic effects with aldosterone were observed. Dehydration and vasopressin infusion increased AQP-2 expression in distal colonic mucosa (P < 0.05). This action of vasopressin was prevented by tolvaptan, a specific V2 receptor antagonist (P < 0.05). It is concluded that vasopressin has trophic effects in the rat distal colon, increasing pericryptal myofibroblast growth which affects crypt absorption, and these effects are independent of the presence of aldosterone. PMID- 17082235 TI - Sulfonylureas and glinides exhibit peroxisome proliferator-activated receptor gamma activity: a combined virtual screening and biological assay approach. AB - Most drugs currently employed in the treatment of type 2 diabetes either target the sulfonylurea receptor stimulating insulin release (sulfonylureas, glinides), or target the peroxisome proliferator-activated receptor (PPARgamma) improving insulin resistance (thiazolidinediones). Our work shows that sulfonylureas and glinides additionally bind to PPARgamma and exhibit PPARgamma agonistic activity. This activity was predicted in silico by virtual screening and confirmed in vitro in a binding assay, a transactivation assay, and by measuring the expression of PPARgamma target genes. Among the measured compounds, gliquidone and glipizide (two sulfonylureas), as well as nateglinide (a glinide), exhibit PPARgamma agonistic activity at concentrations comparable with those reached under pharmacological treatment. The most active of these compounds, gliquidone, is shown to be as potent as pioglitazone at inducing PPARgamma target gene expression. This dual mode of action of sulfonylureas and glinides may open new perspectives for the molecular pharmacology of antidiabetic drugs, because it provides evidence that drugs can be designed that target both the sulfonylurea receptor and PPARgamma. Targeting both receptors could increase pancreatic insulin secretion and improve insulin resistance. Glinides, sulfonylureas, and other acidified sulfonamides may be promising leads in the development of new PPARgamma agonists. In addition, we provide a unified concept of the PPARgamma binding ability of seemingly disparate compound classes. PMID- 17082234 TI - Subunit-specific modulation of T-type calcium channels by zinc. AB - Zinc (Zn2+) functions as a signalling molecule in the nervous system and modulates many ionic channels. In this study, we have explored the effects of Zn2+ on recombinant T-type calcium channels (CaV3.1, CaV3.2 and CaV3.3). Using tsA-201 cells, we demonstrate that CaV3.2 current (IC50, 0.8 microm) is significantly more sensitive to Zn2+ than are CaV3.1 and CaV3.3 currents (IC50, 80 microm and approximately 160 microm, respectively). This inhibition of CaV3 currents is associated with a shift to more negative membrane potentials of both steady-state inactivation for CaV3.1, CaV3.2 and CaV3.3 and steady-state activation for CaV3.1 and CaV3.3 currents. We also document changes in kinetics, especially a significant slowing of the inactivation kinetics for CaV3.1 and CaV3.3, but not for CaV3.2 currents. Notably, deactivation kinetics are significantly slowed for CaV3.3 current (approximately 100-fold), but not for CaV3.1 and CaV3.2 currents. Consequently, application of Zn2+ results in a significant increase in CaV3.3 current in action potential clamp experiments, while CaV3.1 and CaV3.2 currents are significantly reduced. In neuroblastoma NG 108-15 cells, the duration of CaV3.3-mediated action potentials is increased upon Zn2+ application, indicating further that Zn2+ behaves as a CaV3.3 channel opener. These results demonstrate that Zn2+ exhibits differential modulatory effects on T-type calcium channels, which may partly explain the complex features of Zn2+ modulation of the neuronal excitability in normal and disease states. PMID- 17082236 TI - Enhanced aldosterone signaling in the early nephropathy of rats with metabolic syndrome: possible contribution of fat-derived factors. AB - Metabolic syndrome is an important risk factor for proteinuria and chronic kidney disease independent of diabetes and hypertension; however, the underlying mechanisms have not been elucidated. Aldosterone is implicated in target organ injury of obesity-related disorders. This study investigated the role of aldosterone in the early nephropathy of 17-wk-old SHR/NDmcr-cp, a rat model of metabolic syndrome. Proteinuria was prominent in SHR/NDmcr-cp compared with nonobese SHR, which was accompanied by podocyte injury as evidenced by foot process effacement, induction of desmin and attenuation of nephrin. Serum aldosterone level, renal and glomerular expressions of aldosterone effector kinase Sgk1, and oxidative stress markers all were elevated in SHR/NDmcr-cp. Mineralocorticoid receptors were expressed in glomerular podocytes. Eplerenone, a selective aldosterone blocker, effectively improved podocyte damage, proteinuria, Sgk1, and oxidant stress. An antioxidant tempol also alleviated podocyte impairment and proteinuria, along with inhibition of Sgk1. As for the mechanisms of aldosterone excess, visceral adipocytes that were isolated from SHR/NDmcr-cp secreted substances that stimulate aldosterone production in adrenocortical cells. The aldosterone-releasing activity of adipocytes was not inhibited by candesartan. Adipocytes from nonobese SHR did not show such activity. In conclusion, SHR/NDmcr-cp exhibit enhanced aldosterone signaling, podocyte injury, and proteinuria, which are ameliorated by eplerenone or tempol. The data also suggest that adipocyte-derived factors other than angiotensin II might contribute to the aldosterone excess of this model. PMID- 17082237 TI - Role of the Akt/FoxO3a pathway in TGF-beta1-mediated mesangial cell dysfunction: a novel mechanism related to diabetic kidney disease. AB - Diabetic nephropathy (DN) is characterized by mesangial cell (MC) expansion and accumulation of extracellular matrix proteins. TGF-beta is increased in MC under diabetic conditions and in DN and activates key signaling pathways, including the phosphoinositide-3-kinase/Akt (PI3K/Akt) pathway. FoxO transcription factors play roles in cell survival and oxidative stress and are negatively regulated by Akt mediated phosphorylation. We tested whether phosphorylation-mediated inactivation of FoxO3a by TGF-beta can mediate MC survival and oxidative stress. TGF-beta treatment significantly increased levels of p-Akt (activation) and p-FoxO3a (inactivation) in cultured MC. This FoxO3a inactivation was accompanied by significant decreases in the expression of two key FoxO3a target genes, the proapoptotic Bim and antioxidant manganese superoxide dismutase in MC. TGF-beta treatment triggered the nuclear exclusion of FoxO3a, significantly inhibited FoxO3a transcriptional activity, and markedly protected MC from apoptosis. A PI3K inhibitor blocked these TGF-beta effects. It is interesting that p-Akt and p FoxO3A levels also were increased in renal cortical tissues from rats and mice at 2 wk after the induction of diabetes by streptozotocin, thus demonstrating in vivo significance. In summary, TGF-beta and diabetes can increase FoxO3a phosphorylation and transcriptional inactivation via PI3K/Akt. These new results suggest that Akt/FoxO pathway regulation may be a novel mechanism by which TGF beta can induce unopposed MC survival and oxidant stress in early DN, thereby accelerating renal disease. PMID- 17082238 TI - Sleep apnea in patients on conventional thrice-weekly hemodialysis: comparison with matched controls from the Sleep Heart Health Study. AB - Sleep-disordered breathing (SDB) has been noted commonly in hemodialysis (HD) patients, but it is not known whether this is related directly to the treatment of kidney failure with HD or to the higher prevalence of obesity and older age. Forty-six HD patients were compared with 137 participants from the Sleep Heart Health Study (SHHS) who were matched for age, gender, body mass index (BMI), and race. Home unattended polysomnography was performed and scored using similar protocols. The study sample was 62.7 +/- 10.1 yr, was predominantly male (72%) and white (63%), and had an average BMI of 28 +/- 5.3 kg/m(2). The HD sample had a higher systolic BP (137 versus 121 mmHg; P < 0.01) and a higher prevalence of diabetes (33 versus 9%; P < 0.01) and cardiovascular disease (33 versus 13%; P < 0.01) compared with the SHHS sample. The HD group had significantly less sleep time (320 versus 379 min; P < 0.0001) but similar sleep efficiency. HD patients had a higher frequency of arousals per hour (25.1 versus 17.1; P < 0.0001) and apnea-hypopneas per hour (27.2 versus 15.2; P < 0.0001) and greater percentage of the total sleep time below an oxygen saturation of 90% (7.2 versus 1.8; P < 0.0001). HD patients were more likely to have severe SDB (>30 respiratory events per hour) compared with the SHHS sample (odds ratio 4.07; 95% confidence interval 1.83 to 9.07). There was a strong association of HD with severe SDB and nocturnal hypoxemia independent of age, BMI, and the higher prevalence of chronic disease. The potential mechanisms for the higher likelihood of SDB in the HD population must be identified to provide specific prevention and therapy. PMID- 17082239 TI - Endothelin-a receptor blockade improves renal microvascular architecture and function in experimental hypercholesterolemia. AB - Hypercholesterolemia (HC) may trigger early renal injury, partly by impairing the function or the structure of renal microvessels (MV). The endothelin (ET) system is upregulated in HC and can have an impact on the renal microcirculation by regulating MV tone, growth factors, and remodeling. It was hypothesized that ET-A blockade would protect the HC kidney by improving the function and attenuating the damage of intrarenal MV. Single-kidney function and hemodynamic responses to endothelium-dependent challenge were assessed in pigs after 12 wk of experimental HC, HC and chronic supplementation with the ET receptor A blocker ABT-627 (HC+ET A, 0.75 mg/kg per d), and normal controls. Renal MV architecture then was studied ex vivo using three-dimensional microcomputed tomography imaging, and growth factors and remodeling pathways were explored in renal tissue. The HC kidney showed increased MV density compared with normal (77.68 +/- 5.1 versus 62.9 +/- 4.8 vessels/cm(2); P = 0.04) but blunted endothelial function. Chronic ET-A blockade in HC upregulated renal vascular growth factors, further increased renal MV density (139.9 +/- 8.4 vessels/cm(2); P = 0.001 versus normal and HC), and decreased renal tissue and MV remodeling. Furthermore, ET-A blockade in HC decreased MV tortuosity and improved MV endothelial function, suggesting accelerated stabilization and maturation of neo-vessels. Modulation of renal MV architecture and function in HC is mediated partly by the endogenous ET system. Notably, ET-A blockade enhanced the proliferation and facilitated the maturation of renal MV in the HC kidney and improved renal MV remodeling and function. This study suggests novel renoprotective effects of ET-A blockers and supports further exploration of strategies that target the ET pathway in HC and atherosclerosis. PMID- 17082240 TI - Impact of blood pressure control and angiotensin-converting enzyme inhibitor therapy on new-onset microalbuminuria in type 2 diabetes: a post hoc analysis of the BENEDICT trial. AB - For assessment of the independent renoprotective effect of BP control and angiotensin-converting enzyme inhibitor (ACEi) therapy, the relationships of baseline BP, BP reduction, and follow-up BP with the incidence of persistent microalbuminuria were evaluated in 1204 hypertensive patients who had type 2 diabetes and normoalbuminuria and were included in the BErgamo Nephrologic Diabetic Complications Trial (BENEDICT) study and were randomly assigned to 3.6 yr of treatment with the ACEi trandolapril (2 mg/d), the nondihydropyridine calcium channel blocker (ndCCB) verapamil SR (240 mg/d), their fixed combination Veratran (trandolapril 2 mg/d plus verapamil SR 180 mg/d), or placebo, plus other antihypertensive medications targeted at systolic/diastolic BP <130/80 mmHg. Follow-up (from month 3 to study end) systolic, diastolic, mean, and pulse BP and their reductions versus baseline--but not baseline BP--independently predicted (P < 0.001) the risk for microalbuminuria. In patients with follow-up BP above medians, ACEi significantly reduced the risk for microalbuminuria to levels that were observed among patients with BP below medians, regardless of ACEi treatment. The same trend was observed among patients with BP reductions below medians. ndCCB therapy did not independently affect microalbuminuria. Patients who were on Veratran had lower BP and less frequently received diuretics, beta blockers, or dihydropyridine dCCB. In hypertensive, normoalbuminuric patients with type 2 diabetes, BP reduction and ACEi therapy both independently may prevent microalbuminuria. ACEi therapy is particularly effective when BP is poorly controlled, whereas ndCCB therapy is ineffective at any level of achieved BP. As compared with trandolapril, Veratran may help with achievement of target BP with less need for concomitant antihypertensive medications. PMID- 17082241 TI - Calpain activation and secretion promote glomerular injury in experimental glomerulonephritis: evidence from calpastatin-transgenic mice. AB - Glomerular injury and albuminuria in acute glomerulonephritis are related to the severity of inflammatory process. Calpain, a calcium-activated cysteine protease, has been shown to participate in the development of the inflammatory process. Therefore, for determination of the role of calpain in the pathophysiology of acute glomerulonephritis, transgenic mice that constitutively express high levels of calpastatin, a calpain-specific inhibitor protein, were generated. Wild-type mice that were subjected to anti-glomerular basement membrane nephritis exhibited elevated levels of calpain activity in kidney cortex at the heterologous phase of the disease. This was associated with the appearance in urine of calpain activity, which originated potentially from inflammatory cells, abnormal transglomerular passage of plasma proteins, and tubular secretion. In comparison with nephritic wild-type mice, nephritic calpastatin-transgenic mice exhibited limited activation of calpain in kidney cortex and limited secretion of calpain activity in urine. This was associated with less severe glomerular injury (including capillary thrombi and neutrophil activity) and proteinuria. There was a reduction in NF-kappaB activation, suggesting that calpain may participate in inflammatory lesions through NF-kappaB activation. There also was a reduction in nephrin disappearance from the surface of podocytes, indicating that calpain activity would enhance proteinuria by affecting nephrin expression. Exposure of cultured podocytes to calpain decreased nephrin expression, and, conversely, exposure of these cells to calpastatin prevented TNF-alpha from decreasing nephrin expression, demonstrating a role for the secreted form of calpain. Thus, both activation and secretion of calpains participate in the development of immune glomerular injury. PMID- 17082242 TI - Paricalcitol attenuates renal interstitial fibrosis in obstructive nephropathy. AB - Deficiency in vitamin D and its active metabolites is a pathologic feature of chronic kidney diseases. Despite that tubular epithelial cells are the major sites of active vitamin D synthesis, little is known about the role of vitamin D in maintaining the structural and functional integrity of tubular epithelium. This study investigated the effects of paricalcitol (19-nor-1,25-hydroxy-vitamin D(2)), a synthetic vitamin D analogue, on obstructive nephropathy, a model that is characterized by predominant tubulointerstitial lesions. Compared with vehicle controls, paricalcitol significantly attenuated renal interstitial fibrosis in mouse kidney after ureteral obstruction, as demonstrated by a reduced interstitial volume, decreased collagen deposition, and repressed mRNA expression of fibronectin and type I and type III collagens. Paricalcitol largely preserved E-cadherin and reduced alpha-smooth muscle actin expression in vivo. In addition, paricalcitol suppressed renal TGF-beta1 and its type I receptor expression, restored vitamin D receptor abundance, and inhibited cell proliferation and apoptosis after obstructive injury. In vitro, paricalcitol abolished TGF-beta1 mediated E-cadherin suppression and alpha-smooth muscle actin and fibronectin induction in tubular epithelial cells, underscoring its ability to block directly the epithelial to mesenchymal transition (EMT). It is interesting that paricalcitol almost completely suppressed renal induction of Snail, a critical transcription factor that is implicated in EMT programming. Furthermore, paricalcitol inhibited the TGF-beta1-mediated Snail induction in vitro, and ectopic expression of Snail repressed E-cadherin promoter activity and downregulated E-cadherin expression in tubular epithelial cells. These studies suggest that paricalcitol is able to ameliorate renal interstitial fibrosis in obstructive nephropathy, possibly by preserving tubular epithelial integrity through suppression of EMT. PMID- 17082243 TI - African ancestry, socioeconomic status, and kidney function in elderly African Americans: a genetic admixture analysis. AB - Kidney disease is a major public health problem in the United States that affects African Americans disproportionately. The relative contribution of environmental and genetic factors to the increased burden of kidney disease among African Americans is unknown. The associations of genetic African ancestry and socioeconomic status with kidney function were studied cross-sectionally and longitudinally among 736 community-dwelling African Americans who were aged >65 yr and participating in the Cardiovascular Health Study. Genetic African ancestry was determined by genotyping 24 biallelic ancestry-informative markers and combining this information statistically to generate an estimate of ancestry for each individual. Kidney function was evaluated by cystatin C and estimated GFR (eGFR) using the Modification of Diet in Renal Disease equation. Longitudinal changes in serum creatinine and eGFR were estimated using baseline and follow-up values. In cross-sectional analyses, there was no association between genetic African ancestry and either measure of kidney function (P = 0.36 for cystatin C and 0.68 for eGFR). African ancestry was not associated with change in serum creatinine > or =0.05 mg/dl per yr (odds ratio [OR] 0.94; 95% confidence interval [CI] 0.83 to 1.06) or with change in eGFR > or =3 ml/min per 1.73 m(2) per yr (OR 1.02; 95% CI 0.92 to 1.13). In contrast, self reported African-American race was strongly associated with increased risk for kidney disease progression compared with white individuals for change in creatinine (OR 1.77; 95% CI 1.33 to 2.36) and for change in eGFR (OR 3.21; 95% CI 2.54 to 4.06). Among self-identified African Americans, low income (< US dollars 8000/yr) was strongly associated with prevalent kidney dysfunction by cystatin C >1.29 g/dl (adjusted OR 2.7; 95% CI 1.0 to 7.5) or by eGFR <60 ml/min per 1.73 m(2) (adjusted OR 3.2; 95% CI 1.1 to 9.4) compared with those with incomes >US dollars 35,000/yr. Alleles that are known to be present more frequently in the African ancestral group were not associated with kidney dysfunction or kidney disease progression. Rather, kidney dysfunction in elderly African Americans seems more attributable to differences in environmental and social factors. PMID- 17082244 TI - A murine model of HUS: Shiga toxin with lipopolysaccharide mimics the renal damage and physiologic response of human disease. AB - Hemolytic uremic syndrome (HUS), which is caused by Shiga toxin-producing Escherichia coli infection, is the leading cause of acute renal failure in children. At present, there is no complete small animal model of this disease. This study investigated a mouse model using intraperitoneal co-injection of purified Shiga toxin 2 (Stx2) plus LPS. Through microarray, biochemical, and histologic analysis, it was found to be a valid model of the human disease. Biochemical and microarray analysis of mouse kidneys revealed the Stx2 plus LPS challenge to be distinct from the effects of either agent alone. Microarrays identified differentially expressed genes that were demonstrated previously to play a role in this disease. Blood and serum analysis of these mice showed neutrophilia, thrombocytopenia, red cell hemolysis, and increased serum creatinine and blood urea nitrogen. In addition, histologic analysis and electron microscopy of mouse kidneys demonstrated glomerular fibrin deposition, red cell congestion, microthrombi formation, and glomerular ultrastructural changes. It was established that this C57BL/6 mouse is a complete model of HUS that includes the thrombocytopenia, hemolytic anemia, and renal failure that define the human disease. In addition, a time course of HUS disease progression that will be useful for identification of therapeutic targets and development of new treatments for HUS is described. PMID- 17082245 TI - Medical costs of untreated anemia in elderly patients with predialysis chronic kidney disease. AB - The objective of this study was to quantify the incremental medical costs that are associated with untreated anemia among elderly patients with predialysis chronic kidney disease (CKD). An analysis of claims and laboratory data between January 1999 and February 2005 was conducted. Inclusion criteria were age >/=65 yr, two or more hemoglobin readings, one or more claims for CKD, and two or more GFR values of <60 ml/min per 1.73 m(2) (stages 3 to 5 CKD). Patients were excluded when they had cancer or lupus, had received organ transplantation, or were treated for anemia. An open-cohort design was used to classify patients' observation periods into anemia and nonanemia. Both univariate and multivariate analyses were conducted to compare periods of anemia and nonanemia for average monthly medical costs; the latter was adjusted for age, gender, GFR, diabetes, hypertension, liver cirrhosis, coronary artery disease, myocardial infarction, and left ventricular hypertrophy. A subset analysis of patients with moderate CKD (stage 3) was conducted. A total of 2001 patients were identified. Untreated anemia was associated with a significant increase in medical costs, with an unadjusted incremental monthly cost of $1089 (P < 0.0001) and a cost ratio of 1.8:1 relative to nonanemia. After controlling for covariates, untreated anemia remained significantly associated with a cost increase (adjusted incremental monthly cost $503; cost ratio 1.4:1; P < 0.0001). Similar significant cost burden was observed in the subset of patients with moderate CKD. The retrospective observational design may be more susceptible to bias than a randomized, controlled trial. This large study, which was based on real-life practice data, demonstrated that untreated anemia in elderly patients with predialysis CKD was associated with a significant increase in medical costs. PMID- 17082246 TI - Ligands to nucleic acid-specific toll-like receptors and the onset of lupus nephritis. AB - Lupus nephritis develops from a combination of genetic and environmental factors such as microbial infection. A role for microbial nucleic acids (e.g., via nucleic acid-specific Toll-like receptors [TLR]) was hypothesized, in this context, because microbial nucleic acids can trigger multiple aspects of autoimmunity in vitro and in vivo. Eight-week-old MRL(lpr/lpr) and MRL wild-type mice received an injection of pI:C RNA (ligand to TLR-3), imiquimod (ligand to TLR-7), or CpG-DNA (ligand to TLR-9) on alternate days for 2 wk. Only CpG-DNA triggered the onset of lupus nephritis in MRL(lpr/lpr) mice, as defined by diffuse proliferative glomerulonephritis associated with glomerular IgG and complement C3 deposition, proteinuria, and glomerular macrophage infiltrates. None of the compounds caused DNA autoantibody production or glomerulonephritis in MRL wild-type mice. The role of CpG-DNA to trigger lupus nephritis in MRL(lpr/lpr) mice was found to relate to its potent immunostimulatory effects at multiple levels: B cell IL12p40 production, B cell proliferation, double-stranded DNA autoantibody secretion, and dendritic cell IFN-alpha production. The induction of lupus nephritis by CpG-DNA is motif specific and could be prevented by co-injection of inhibitory DNA. In summary, among the ligands tested, CpG-DNA triggers lupus nephritis in genetically predisposed hosts. These data support the concept that systemic lupus erythematosus is triggered by pathogens that release CG-rich DNA. PMID- 17082247 TI - Improved marker combination for detection of de novo genetic variation and aberrant DNA in colorectal neoplasia. AB - BACKGROUND: The genetic heterogeneity of sporadic colorectal cancer (CRC) makes the choice of genetic markers and sequence variation-detection technologies critical to the performance of screening assays. We have previously described the effectiveness of a CRC assay composed of 22 known variants in KRAS, APC, TP53, and BAT-26 (V1). We introduce a new marker formulation (V2) that includes detection of de novo variation in APC, PIK3CA, and CTNNB1, hypermethylated sequences within SMARCA3 and VIM, and a single-base variation within BRAF. We compared the abilities of the V1 and V2 markers to detect aberrant DNA in colorectal neoplasias. METHODS: V1 and V2 marker formulations were used to analyze 144 colorectal tissue samples comprising 50 precancerous adenomas, 94 carcinomas, and 11 nonpathologic tissues. V1 analysis consisted of single-base extension analysis of the 22 V1 variants. V2 analysis consisted of DNA scanning of the APC mutation cluster region, PIK3CA exons 9 and 20, CTNNB1 exon 3, analysis for the BRAF Val600Glu substitution, and methylation-specific PCR analysis of VIM and SMARCA3. RESULTS: The V2 marker formulation had significantly higher sensitivity than the V1 markers for carcinomas (93.6% and 72.3%, respectively; P = 0.0002) and adenomas (92.0% and 62.0%, respectively; P = 0.0006). None of the nonpathologic samples were positive for any marker. CONCLUSIONS: We demonstrate improved sensitivity of a new marker formulation (V2) to detect aberrant DNA in CRC and precancerous adenoma tumor tissues. PMID- 17082248 TI - Tandem mass spectrometry for the direct assay of enzymes in dried blood spots: application to newborn screening for mucopolysaccharidosis II (Hunter disease). AB - BACKGROUND: A treatment for mucopolysaccharidosis II (Hunter syndrome) has recently become available. Therefore, we developed a high-throughput assay method appropriate for newborn screening for the relevant enzyme, iduronate 2-sulfatase. METHODS: We synthesized a new iduronate 2-sulfatase substrate that can be used to assay the enzyme by use of tandem mass spectrometry together with an internal standard. The assay uses a dried blood spot on a newborn screening card as the enzyme source. RESULTS: When the assay was tested on dried blood spots, the iduronate 2-sulfatase activity measured for 13 patients with Hunter syndrome was well below the interval found for 57 randomly chosen newborns. The assay was more sensitive than previously reported iduronate 2-sulfatase assays. CONCLUSIONS: This newly developed tandem mass spectrometry assay has the potential to be adopted for newborn screening of Hunter syndrome. This method also has the potential to be carried out in multiplex fashion to assay several different enzymes relevant to lysosomal storage diseases that are assayed in a single infusion into the mass spectrometer. PMID- 17082249 TI - MALDI-TOF mass spectrometry for multiplex genotyping of CYP2B6 single-nucleotide polymorphisms. AB - BACKGROUND: CYP2B6 is a highly variable and polymorphic cytochrome P450 (CYP) enzyme involved in the biotransformation of an increasing number of drugs, including cyclophosphamide, bupropion, and the nonnucleosidic reverse transcriptase inhibitor efavirenz. Several nonsynonymous and promoter single nucleotide polymorphisms (SNPs) in the CYP2B6 gene are associated with altered hepatic expression and function, which affect drug plasma concentrations. METHODS: We used multiplex PCR to amplify relevant gene fragments while avoiding amplification of the CYP2B7P1 pseudogene. Polymorphic sites were analyzed by allele-specific primer extension followed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Method evaluation was performed on a panel of 287 genomic DNA samples previously genotyped by other methods. RESULTS: Five multiplex assays were developed, comprising the following 15 SNPs: -82T --> C (*22); 86G --> C (R29T, *17); 136A - > G (M46V, *11); 296G --> A (G99E, *12); 415A --> G (K139E, *8, *13); 419G --> A (R140Q, *14); 516G --> T (Q172H, *6, *7, *9, *13, *19, *20), 547G --> A (V183I); 769G --> A (D257N); 785A --> G (K262R, *4, *6, *7, *13, *16, *19, *20); 983T-->C (I328T, *16, *18); 1006C --> T (R336C, *19); 1172T --> A (I391N, *15); 1282C --> A (P428T, *21); 1459C --> T (R487C, *5, *7). In 9 DNA samples showing discrepant genotypes, correctness of the MALDI-TOF MS result was confirmed by direct sequencing. CONCLUSIONS: This genotyping method enabled sensitive, specific, accurate, and comprehensive determination of 15 relevant SNPs of CYP2B6. The assay design allows analysis of SNP subsets, incorporation of additional SNPs, and performance of high-throughput genotyping. PMID- 17082250 TI - Pax2 gene dosage influences cystogenesis in autosomal dominant polycystic kidney disease. AB - Mutations in PKD1 cause dominant polycystic kidney disease (PKD), characterized by large fluid-filled kidney cysts in adult life, but the molecular mechanism of cystogenesis remains obscure. Ostrom et al. [Dev. Biol., 219, 250-258 (2000)] showed that reduced dosage of Pax2 caused increased apoptosis, and ameliorated cystogenesis in Cpk mutant mice with recessive PKD. Pax2 is expressed in condensing metanephrogenic mesenchyme and arborizing ureteric bud, and plays an important role in kidney development. Transient Pax2 expression during fetal kidney mesenchyme-to-epithelial transition, as well as in nascent tubules, is followed by marked down-regulation of Pax2 expression. Here, we show that in humans with PKD, as well as in Pkd1(del34/del34) mutant mice, Pax2 was expressed in cyst epithelial cells, and facilitated cyst growth in Pkd1(del34/del34) mutant mice. In Pkd1(del34/del34) mutant kidneys, the expression of Pax2 persisted in nascent collecting ducts. In contrast, homozygous Pkd1(del34/del34) fetal mice carrying mutant Pax2 exhibited ameliorated cyst growth, although reduced cystogenesis was not associated with increased apoptosis. Pax2 expression was attenuated in nascent collecting ducts and absent from remnant cysts of Pkd1(del34/del34)/Pax2(1Neu/+) mutant mice. To investigate whether the Pkd1 gene product, Polycystin-1, regulates Pax2, MDCK cells were engineered constitutively expressing wild-type Pkd1; Pax2 protein levels and promoter activity were both repressed in MDCK cells over-expressing Pkd1, but not in cells without transgenic Pkd1. These data suggest that polycystin-1-deficient tubular epithelia persistently express Pax2 in ADPKD, and that Pax2 or its pathway may be an appropriate target for the development of novel therapies for ADPKD. PMID- 17082251 TI - Intragenic deletion of Tgif causes defectsin brain development. AB - TG-interacting factor (TGIF) is a homeodomain-containing protein and functions as a transcriptional repressor within the TGF-beta and retinoic acid signaling pathways. Heterozygous mutations of TGIF have been found in patients with holoprosencephaly (HPE), which is the most common congenital brain malformation in humans. However, targeted null deletions of the entire Tgif gene in mice surprisingly revealed no apparent brain defects. We report here that deletion of the third exon of Tgif gene resulted in a defined spectrum of brain developmental defects including exencephaly, microcephaly, HPE, and abnormalities in embryonic brain ventricle formation and cleavage. These defects could be detected in mice both heterozygous and homozygous for the targeted Tgif deletion. Moreover, expression of dorsal-ventral patterning genes including Shh, Pax6 and Nkx2.2 was altered. The ventricular neuroepithelium exhibited focalized increase of cell proliferation rate and resultant tissue expansion. The incidence of brain abnormalities within the mutant mice was dependent on its genetic background, suggesting that additional genetic modifiers functionally interact with Tgif during embryonic brain development. The intragenic Tgif deletion mouse, therefore, would serve as a useful model that can be used to unravel the genetic complexity implicated in the pathogenesis of HPE. PMID- 17082252 TI - Prediction of antitumour necrosis factor clinical efficacy by real-time visualisation of apoptosis in patients with Crohn's disease. AB - BACKGROUND: The human anti-tumour necrosis factor (TNF) antibody infliximab binds to the membrane TNF and subsequently induces apoptosis of activated lamina propria T lymphocytes in patients with Crohn's disease in vitro. AIM: To test whether the ability of rapid anti-TNF-induced apoptosis in the gut predicts the efficacy of anti-TNF treatment in inflammatory bowel disease. METHODS: (99m)Technetium-annexin V single-photon emission computer tomography (SPECT) was performed in 2 models of murine experimental colitis and in 14 patients with active Crohn's disease as assessed by the Crohns Disease Activity Index (CDAI) to study the effect of anti-TNF treatment on apoptosis in the intestine during active colitis. Disease activity was evaluated 2 weeks after infliximab infusion using the CDAI (definition response: drop of >100 points). RESULTS: Colonic uptake of (99m)Tc-annexin V significantly increased in 2,4,6-trinitrobenzene sulphonate-induced colitis as well as in transfer colitis on administration of anti-TNF antibodies compared with a control antibody as determined with dedicated animal pinhole SPECT. In addition, uptake of (99m)Tc-annexin V significantly increased in patients with active Crohn's disease responding to infliximab treatment. Colonic (99m)Tc-annexin V uptake ratio (mean (SEM)) increased from 0.24 (0.03) to 0.41(0.07) (p<0.01), 24 h after infliximab infusion (5 mg/kg). A mean increase of 98.7% in colonic uptake of (99m)Tc-annexin V could be detected in 10 of the 14 responding patients (CDAI >100 points at week 2) compared with 15.2% in non-responding patients (p = 0.03). Analysis of the mucosal biopsy specimens identified lamina propria T cells as target cells undergoing apoptosis. CONCLUSIONS: These in vivo observations support the notion that colonic uptake of (99m)Tc-annexin V correlates with clinical benefit of anti-TNF treatment and might be predictive of therapeutic success. PMID- 17082253 TI - 17beta-estradiol protects cortical neurons against oxidative stress-induced cell death through reduction in the activity of mitogen-activated protein kinase and in the accumulation of intracellular calcium. AB - Although many studies have suggested that estrogen acts as a neuroprotective agent in oxidative stress, the underlying mechanism has not been fully elucidated. In the present study, we examined the effect of 17beta-estradiol (17beta-E2) on H(2)O(2)-induced death signaling in cultured cortical neurons. Exposure of the cortical neurons to H(2)O(2) triggered a series of events, including overactivation of p44/42 MAPK and intracellular Ca(2+) accumulation via voltage-gated Ca(2+) channels and ionotropic glutamate receptors, resulting in apoptotic-like cell death. The MAPK pathway might work as death signaling in our system, because the MAPK pathway inhibitor, U0126, blocked H(2)O(2)-induced MAPK activation, Ca(2+) overload, and cell death. Interestingly, a similar inhibitory effect on H(2)O(2)-triggered MAPK activation, Ca(2+) accumulation, and cell death was observed in cultures incubated with 17beta-E2 for 24 h before exposure to H(2)O(2), suggesting that the protective effect of 17beta-E2 is induced via attenuating overactivation of the MAPK pathway. Furthermore, we found that ionotropic glutamate receptor subunits, including NR2A and GluR2/3, but not NR2B and GluR1, were down-regulated in the 17beta-E2-treated cultures. The down regulation of these glutamate receptor subunits was also observed after chronic treatment with U0126. Therefore, it is possible that 17beta-E2 down-regulates the expression of the ionotropic glutamate receptors by reducing activity of the MAPK pathway, which might be important for the protective effect of 17beta-E2 against oxidative stress-induced toxicity. PMID- 17082254 TI - The role of inducible repressor proteins in the adrenergic induction of arylalkylamine-N-acetyltransferase and mitogen-activated protein kinase phosphatase-1 in rat pinealocytes. AB - In this study, we investigated the role of two inducible repressor proteins, inducible cAMP early repressor (ICER) and Fos-related antigen 2 (Fra-2) in the adrenergic induction of MAPK phosphatase-1 (MKP-1) as compared with their roles in the induction of arylalkylamine-N-acetyltransferase (AA-NAT) in rat pinealocytes. Treatment of pinealocytes with norepinephrine (NE) caused an increase in the mRNA and protein levels of MKP-1 and AA-NAT, as well as in the AA NAT activity and melatonin production. NE stimulation also caused a simultaneous increase in the mRNA and protein levels of ICER and Fra-2. Transient knockdown of icer using adenovirus expressing small interfering RNA (siRNA) abolished the NE induction of icer expression but had little effect on the NE induction of mkp-1 or aa-nat expression. In contrast, pretreatment with adenovirus overexpressing icer was effective in reducing the NE induction of mkp-1 and aa-nat. The inhibitory effect of overexpressing icer was reversed by cotreatment with siRNA against icer. siRNA against fra-2 also abolished the NE-stimulated expression of fra-2 but had little effect on the NE induction of mkp-1 and aa-nat expression. Proteasomal inhibition, which reduced the NE-stimulated induction of aa-nat, caused a reduction of ICER and Fra-2. Together, these results indicate that whereas overexpression of ICER can suppress the NE induction of aa-nat and mkp-1, the amount of the repressors, ICER and Fra-2, present during NE induction appears insufficient to exert a significant effect in controlling the expression of these genes. PMID- 17082255 TI - Astressin B, a corticotropin-releasing hormone receptor antagonist, accelerates the return to normal luteal function after an inflammatory-like stress challenge in the rhesus monkey. AB - Endogenous release of CRH in stress has been associated with a dysfunctional reproductive endocrine axis. In the rhesus monkey, an inflammatory-like stress challenge in the luteal phase decreases luteal secretory function. Here, we tested the effectiveness of astressin B, a nonspecific CRH receptor antagonist, in constraining the deleterious impact of a 10-d lipopolysaccharide (LPS) challenge on the menstrual cycle. Two protocols were carried out in nine animals. In the first, the animals, after showing two normal consecutive control cycles, were injected daily for 10 days with LPS (75-125 mug/d) during the luteal phase of the cycle. The animals were followed through the two postchallenge cycles. The second protocol, carried out in the following year, was identical with protocol 1, except that the animals were treated with astressin B (0.45 mg/kg) 1 h before each daily LPS challenge during the luteal phase. Blood samples were obtained daily to document cyclic hormones levels. The LPS challenge significantly decreased luteal progesterone and LH release during the challenge cycle. Inhibition of luteal progesterone extended to the two successive postchallenge cycles. Astressin B treatment prevented luteal LH but not luteal progesterone decrease during the treatment cycle and restored normal progesterone secretion during the two posttreatment cycles. We conclude that the deleterious impact of a short-term inflammatory stress challenge on luteal function is far longer than the stress period itself. Systemic administration of astressin B accelerates the return to normal luteal function, presumably by restoring normal neuroendocrine regulation of gonadotropin secretion. PMID- 17082256 TI - Vasopressin autoreceptors and nitric oxide-dependent glutamate release are required for somatodendritic vasopressin release from rat magnocellular neuroendocrine cells responding to osmotic stimuli. AB - Magnocellular neuroendocrine cells of the supraoptic nucleus (SON) release vasopressin (VP) systemically and locally during osmotic challenge. Although both central VP and nitric oxide (NO) release appear to reduce osmotically stimulated systemic VP release, it is unknown whether they interact locally in the SON to enhance somatodendritic release of VP, a phenomenon believed to regulate systemic VP release. In this study, we examined the contribution of VP receptor subtypes and NO to local VP release from the rat SON elicited by systemic injection of 3.5 m saline. Treatment of SON punches with VP receptor antagonists decreased osmotically stimulated intranuclear VP release. Similarly, blockade of NO production, or addition of NO scavengers, reduced stimulated VP, glutamate, and aspartate release, suggesting that local NO production and activity are critical for osmotically induced intranuclear VP and excitatory amino acid release. An increase in endogenous NO release from SON punches in response to hyperosmolality was confirmed by enzymatic NO assay. Consistent with enhanced glutamate and VP release from stimulated rat SON punches, the ionotropic glutamate receptor blocker kynurenate decreased stimulated local VP release without affecting NO release. These data suggest that NO enhances local VP release in part by facilitating local release of glutamate/aspartate and that glutamate receptor activity is required for the stimulation of local VP release by osmotic challenge. Collectively, these results suggest that local VP receptors, NO, and glutamatergic signaling mediate the amplification of intranuclear VP release during hyperosmolality and may contribute to efficient, but not exhaustive, systemic release of VP during osmoregulatory challenge. PMID- 17082257 TI - Steroid and G protein binding characteristics of the seatrout and human progestin membrane receptor alpha subtypes and their evolutionary origins. AB - A novel progestin receptor (mPR) with seven-transmembrane domains was recently discovered in spotted seatrout and homologous genes were identified in other vertebrates. We show that cDNAs for the mPR alpha subtypes from spotted seatrout (st-mPRalpha) and humans (hu-mPRalpha) encode progestin receptors that display many functional characteristics of G protein-coupled receptors. Flow cytometry and immunocytochemical staining of whole MDA-MB-231 cells stably transfected with the mPRalphas using antibodies directed against their N-terminal regions show the receptors are localized on the plasma membrane and suggest the N-terminal domain is extracellular. Both recombinant st-mPRalpha and hu-mPRalpha display high affinity (Kd 4.2-7.8 nm), limited capacity (Bmax 0.03-0.32 nm), and displaceable membrane binding specific for progestins. Progestins activate a pertussis toxin sensitive inhibitory G protein (G(i)) to down-regulate membrane-bound adenylyl cyclase activity in both st-mPRalpha- and hu-mPRalpha-transfected cells. Coimmunoprecipitation experiments demonstrate the receptors are directly coupled to the G(i) protein. Similar to G protein-coupled receptors, dissociation of the receptor/G protein complex results in a decrease in ligand binding to the mPRalphas and mutation of the C-terminal, and third intracellular loop of st mPRalpha causes loss of ligand-dependent G protein activation. Phylogenetic analysis indicates the mPRs are members of a progesterone and adipoQ receptor (PAQR) subfamily that is only present in chordates, whereas other PAQRs also occur in invertebrates and plants. Progesterone and adipoQ receptors are related to the hemolysin3 family and have origins in the Eubacteria. Thus, mPRs arose from Eubacteria independently from members of the GPCR superfamily, which arose from Archeabacteria, suggesting convergent evolution of seven-transmembrane hormone receptors coupled to G proteins. PMID- 17082258 TI - Induction of cardiac uncoupling protein-2 expression and adenosine 5' monophosphate-activated protein kinase phosphorylation during early states of diet-induced obesity in mice. AB - The objective of this work was to characterize the adaptation of cardiac metabolism to a lipid overload in a model of diet-induced obesity (DIO) in mice. After 8 wk dietary treatment, mice receiving a high-fat diet exhibited an increase in the amount of adipose tissue, accompanied by a surge in plasma leptin concentration (from 5.4-16.0 ng/ml). This was associated with: 1) an induction of uncoupling protein-2 (120%), 2) an increase in the phosphorylated form of AMP activated protein kinase (120%), and 3) a reduction in lactate concentration and lactate dehydrogenase activity in myocardial tissue (40%). Because DIO induces leptin resistance, we analyzed leptin receptor functionality by measuring phospho signal transducer and activator of transcription 3 in response to acute leptin (1 mg/kg). We observed that leptin receptor signaling remained unaltered within the heart but was fully impaired within the hypothalamus. Taken together, these data show that during DIO development, there is a metabolic shift in the heart aimed at increasing fatty acid oxidation to the detriment of carbohydrates. This effect seems to be leptin-dependent, suggesting that the increased adiposity observed during the onset of obesity might contribute to impairing ectopic lipidic deposition in the heart. PMID- 17082259 TI - Macrophage-secreted factors impair human adipogenesis: involvement of proinflammatory state in preadipocytes. AB - Obesity is considered a chronic low-grade inflammatory state. The white adipose tissue produces a variety of inflammation-related proteins whose expression is increased in obese subjects. The nonadipose cell fraction, which includes infiltrated macrophages, is a determinant source of inflammation-related molecules within the adipose tissue. Our working hypothesis is that macrophage infiltration affects fat expansion through a paracrine action on adipocyte differentiation. Human primary preadipocytes were then differentiated in the presence of conditioned media obtained from macrophages differentiated from blood monocytes. Preadipocytes treated by macrophage-conditioned medium displayed marked reduction of adipogenesis as assessed by decreased cellular lipid accumulation and reduced gene expression of adipogenic and lipogenic markers. In addition to this effect, the activation of macrophages by lipopolysaccharides stimulated nuclear factor kappaB signaling, increased gene expression and release of proinflammatory cytokines and chemokines, and induced preadipocyte proliferation. This phenomenon was associated with increased cyclin D1 gene expression and maintenance of the fibronectin-rich matrix. Anti-TNFalpha neutralizing antibody inhibits the inflammatory state of preadipocytes positioning TNFalpha as an important mediator of inflammation in preadipocytes. Strikingly, conditioned media produced by macrophages isolated from human adipose tissue exerted comparable effects with activated macrophages, i.e. decreased adipogenesis and increased inflammatory state in the preadipocytes. These data show that macrophage-secreted factors inhibit the formation of mature adipocytes, suggesting possible role in limiting adipose tissue expansion in humans. PMID- 17082260 TI - Pre-B-cell transcription factor 1 and steroidogenic factor 1 synergistically regulate adrenocortical growth and steroidogenesis. AB - A variety of transcription factors including Wilms tumor gene (Wt-1), steroidogenic factor 1 (Sf-1), dosage-sensitive sex reversal, adrenal hypoplasia congenita on the X-chromosome, Gene 1 (Dax-1), and pre-B-cell transcription factor 1 (Pbx1) have been defined as necessary for regular adrenocortical development. However, the role of Pbx1 for adrenal growth and function in the adult organism together with the molecular relationship between Pbx1 and these other transcription factors have not been characterized. We demonstrate that Pbx haploinsufficiency (Pbx1(+/-)) in mice is accompanied by a significant lower adrenal weight in adult animals compared with wild-type controls. Accordingly, baseline proliferating cell nuclear antigen levels are lower in Pbx1(+/-) mice, and unilateral adrenalectomy results in impaired contralateral compensatory adrenal growth, indicating a lower proliferative potential in the context of Pbx1 haploinsufficiency. In accordance with the key role of IGFs in adrenocortical proliferation and development, real-time RT-PCR demonstrates significant lower expression levels of the IGF-I receptor, and up-regulation of IGF binding protein 2. Functionally, Pbx1(+/-) mice display a blunted corticosterone response after ACTH stimulation coincident with lower adrenal expression of the ACTH receptor (melanocortin 2 receptor, Mc2-r). Mechanistically, in vitro studies reveal that Pbx1 and Sf-1 synergistically stimulates Mc2-r promoter activity. Moreover, Sf-1 directly activates the Pbx1 promoter activity in vitro and in vivo. Taken together, these studies provide evidence for a role of Pbx1 in the maintenance of a functional adrenal cortex mediated by synergistic actions of Pbx1 and Sf-1 in the transcriptional regulation of the critical effector of adrenocortical differentiation, the ACTH receptor. PMID- 17082261 TI - Congenital hypothyroid female pax8-deficient mice are infertile despite thyroid hormone replacement therapy. AB - Absence of the Pax8 gene results in congenital hypothyroidism in mice, and mutations of the Pax8 gene have been associated with thyroid hypoplasia in humans. As in humans, treatment of congenital hypothyroid Pax8 null mice with thyroxine normalizes the known deficits. However, we report here that thyroxine substituted female Pax8(-/-) mice are infertile because they lack a functional uterus revealing only remnants of myometrial tissue. In addition, the vaginal opening is absent. Interestingly, oviduct, cervix, and upper parts of the vagina are not affected, although Pax8 expression has been described in the entire Mullerian duct before. Because the natural outflow of the oviduct is impaired, a hydrosalpinx develops frequently. Folliculogenesis, ovarian hormone production, and transcription of pituitary hormones are in a normal range. Thus, infertility in Pax8(-/-) mice seems to be due to a defect in development of the Mullerian duct rather than to hormonal imbalance, pointing to a direct morphogenic role for Pax8 in uterine development. Because we demonstrated Pax8 expression not only in the uterine epithelium of mice but also in the human endometrium, it remains to be elucidated whether adequate development of the uterus may also be affected in congenital hypothyroid female patients with mutations in the Pax8 gene. PMID- 17082262 TI - Glucose activates a protein phosphatase-1-mediated signaling pathway to enhance overall translation in pancreatic beta-cells. AB - Both the rate of overall translation and the specific acceleration of proinsulin synthesis are known to be glucose-regulated processes in the beta-cell. In this study, we propose that glucose-induced stimulation of overall translation in beta cells depends on a protein phosphatase-1-mediated decrease in serine-51 phosphorylation of eukaryotic translation initiation factor 2alpha (eIF2alpha), a pivotal translation initiation factor. The decrease was rapid and detectable within 15 min and proportional to the range of glucose concentrations that also stimulate translation. Lowered net eIF2alpha phosphorylation was not associated with a detectable decrease in activity of any eIF2alpha kinase. Moreover, okadaic acid blocked glucose-induced eIF2alpha dephosphorylation, suggesting that the net effect was mediated by a protein phosphatase. Experiments with salubrinal on intact cells and nuclear inhibitor of protein phosphatase-1 (PP1) on cell extracts suggested that this phosphatase was PP1. The net effect contained, however, a component of glucose-induced folding load in the endoplasmic reticulum because coincubation with cycloheximide further amplified the effect of glucose on eIF2alpha dephosphorylation. Thus, the steady-state level of eIF2alpha phosphorylation in beta-cells is the result of a balance between folding-load induced phosphorylation and PP1-dependent dephosphorylation. Because defects in the pancreatic endoplasmic reticulum kinase-eIF2alpha signaling system lead to beta-cell failure and diabetes, deregulation of the PP1 system could likewise lead to cellular dysfunction and disease. PMID- 17082263 TI - Activin and glucocorticoids synergistically activate follicle-stimulating hormone beta-subunit gene expression in the immortalized LbetaT2 gonadotrope cell line. AB - FSH is produced by the pituitary gonadotrope to regulate gametogenesis. Production of the beta-subunit of FSH is the rate-limiting step in FSH synthesis, and a number of peptide and steroid hormones within the reproductive axis have been found to regulate transcription of the FSH beta-subunit gene. Although both activin and glucocorticoids are notable regulators of FSHbeta by themselves, we find that cotreatment results in a synergistic interaction on the mouse FSHbeta promoter at the level of the gonadotrope using transient transfection of a reporter gene into the LbetaT2 immortalized gonadotrope-derived cell line. This synergistic interaction is specific to FSHbeta, because only additive effects of these two hormones are observed on LH beta-subunit, GnRH receptor, and mouse mammary tumor virus gene expression. Components of both activin and glucocorticoid signaling are found to be necessary for synergy, and there are specific cis elements on the mouse FSHbeta promoter that contribute to the synergistic response as well. We also identify novel activin-responsive regions in the mouse FSHbeta promoter and find that the -120 site can bind Smad2/3 in vitro. In addition, the glucocorticoid receptor and Smad3 are sufficient to confer a striking synergy with glucocorticoids on the mouse FSHbeta promoter. Our studies provide the first evidence of a synergistic interaction between activin and glucocorticoids within the gonadotrope cell and demonstrate that this synergy can occur directly at the level of the mouse FSHbeta promoter. PMID- 17082264 TI - Opprobrium or opportunity? PMID- 17082265 TI - Practicing dentistry using findings from clinical research: you are closer than you think. PMID- 17082266 TI - Do no harm. PMID- 17082267 TI - Kudos to ADA. PMID- 17082268 TI - Nonsplint therapies. PMID- 17082269 TI - Saliva. PMID- 17082270 TI - Findings questioned. PMID- 17082271 TI - An alternative explanation. PMID- 17082272 TI - Oral bisphosphonates and bon. PMID- 17082277 TI - The neuropathology, medical management and dental implications of autism. AB - BACKGROUND: A paucity of information exists in the dental literature about autism and its dental implications. TYPES OF STUDIES REVIEWED: The authors conducted a MEDLINE search for the period 2000 through 2006, using the term "autism," with the aim of defining the condition's clinical manifestations, dental and medical treatment and dental implications. RESULTS: Autism is a severe developmental brain disorder that appears in infancy, persists throughout life, and is characterized by impaired social interaction, abnormalities in communication (both verbal and nonverbal) and restricted interests. Often accompanying the disorder are behavioral disturbances - such as self-mutilation, aggression, psychiatric symptoms and seizures - that necessitate the administration of multiple medications to help the affected person participate effectively in the educational and rehabilitative process. CLINICAL IMPLICATIONS: Dentists caring for people with autism must be familiar with the manifestations of the disease and its associated features so that they can garner the maximum level of patient cooperation. They also must be familiar with the medications used to treat the associated features of the disorder because many of them cause untoward orofacial and systemic reactions and may precipitate adverse interactions with dental therapeutic agents. PMID- 17082278 TI - A clinical evaluation of resin-based composite and glass ionomer cement restorations placed in primary teeth using the ART approach: results at 24 months. AB - BACKGROUND: The authors evaluated the 24-month performance of a packable resin based composite/dentin bonding system and a high-viscosity glass ionomer cement (GIC) in restorations placed in primary molars with the atraumatic restorative treatment (ART) approach. METHODS: Three dentists placed 419 restorations in 219 children aged 6 through 10 years who had bilateral matched pairs of carious posterior Class I and II primary teeth. They used a split-mouth design to place the two materials, which were assigned randomly to contralateral sides. The authors evaluated the restorations according to U.S. Public Health Service Ryge criteria. RESULTS: After 24 months, 96.7 percent of the Class I GIC restorations and 91 percent of the resin-based composite restorations survived, while the success rates for the Class II restorations were 76.1 percent and 82 percent for the GIC and resin-based composite restorations, respectively. The survival rate of the Class II resin-based composite restorations was 5.9 percent higher than that of the GIC restorations at the 24-month evaluation, but this difference was not statistically significant. However, the study results showed a statistically significant difference in survival rates between Class I and II restorations for both materials. CONCLUSION AND CLINICAL IMPLICATIONS: The two-year clinical performance of both materials was satisfactory for the restoration of Class I and II primary molars using the ART approach. PMID- 17082279 TI - A red lesion on the palate. PMID- 17082280 TI - The transmission of BANA-positive periodontal bacterial species from caregivers to children. AB - BACKGROUND: The purpose of the authors' study was to use the N-benzoyl-DL arginine-2-naphthy-lamide (BANA) test (BANAMet LLC, Ann Arbor, Mich.) to obtain information regarding the prevalence of an enzyme unique to certain periodontal pathogens in plaque samples of children, as well as the potential transmission of these pathogens from caregivers to children. METHODS: The authors tested 218 subjects (3 to 10 years old) and 195 care-givers at four pediatric dentistry clinics in Taipei, Taiwan. RESULTS: Forty-four percent of the children had at least one plaque sample that tested positive and/or weakly positive. Positive results were more frequent in the mixed dentition, as well as in children with gingivitis (P < .001). A logistic regression model showed that if the BANA test results for the care-giver were positive, the odds of the child's also having positive test results were 55 times greater (P < .001; confidence interval [CI] = 14 to 224) than those for a child whose caregiver had negative BANA test results. Other predictors were the presence of a mixed dentition (P < .001; odds ratio [OR] = 11; CI = 3.5 to 33.5) and the children's papillary bleeding scores (P < .001, OR = 3.1, CI = 2.0 to 4.7). CONCLUSION: The BANA test results were positive for almost one-half of the children. A positive reaction was associated with gingivitis, a mixed dentition, a BANA-positive caregiver or a caregiver with a history of periodontal disease in the family. CLINICAL IMPLICATIONS: The authors propose an anaerobic periodontal infection risk model in which children with a mixed dentition who have gingivitis and a caregiver with a history of periodontal disease would undergo the BANA test. PMID- 17082281 TI - Third-molar extraction as a risk factor for temporomandibular disorder. AB - BACKGROUND: Although third-molar extraction is common in the U.S., there are few reports on the relationship between third-molar extraction and temporomandibular disorder (TMD). The aim of this study was to investigate this potential association. METHODS: The authors enrolled study subjects on their 15th birthdays and followed them continuously for a minimum of five years. The authors ascertained exposure (third-molar extraction) and outcome (TMD) via electronic dental insurance records. A survival analysis design estimated the relative risk of experiencing TMD after third-molar extraction. They considered sex, dental care utilization and other potentially confounding variables in the analyses. RESULTS: A total of 34,491 subjects met the inclusion criterion. Fifty percent of all subjects had third molars removed by the age of 20 years, and 391 subjects had claims indicating TMD. The adjusted relative risk of experiencing TMD after third-molar extraction was 1.6 (95 percent confidence interval, 1.3 to 2.0). Calculation of the population risk indicated that 23 percent of all TMD cases in this age group might be due to third-molar extraction. CONCLUSIONS: Third-molar extraction appears to be a risk factor for TMD. CLINICAL IMPLICATIONS: Dental providers should be aware of the risk of experiencing TMD related to third-molar extraction and take measures to minimize trauma to the joint during extraction. PMID- 17082282 TI - Three-dimensional imaging using microcomputed tomography for studying tooth macromorphology. AB - BACKGROUND: The authors conducted a study to demonstrate potential applications of microcomputed tomography (microCT) in the analysis of tooth morphology. METHODS: The authors selected for microCT analysis five maxillary first molars with a second canal in the mesiobuccal (MB) root, five mandibular first molars with a mesial root possessing a considerable curvature and five single-canal premolars with complicated apical anatomy. The hardware device used in this study was a desktop X-ray microfocus CT scanner (SkyScan 1072, SkyScan bvba, Aartselaar, Belgium). RESULTS: The authors obtained a three-dimensional image from each of the 15 teeth. In three cases, the MB canals coalesced into one canal, while in the other two molars the canals were separate. Four of the five mandibular molars exhibited a single canal in the mesial root, which had a broad, flat appearance in a mesiodistal dimension. In the premolar teeth, the canals were independent; however, the apical delta and ramifications of the root canals were obvious, yet intricate. CONCLUSIONS: MicroCT offers a reproducible technique for 3-D noninvasive assessment of root canal systems. CLINICAL IMPLICATIONS: While this technique is not suitable for clinical use, it can be applied to improve preclinical training and analysis of fundamental procedures in endodontic and restorative treatment. PMID- 17082283 TI - The pharmacokinetics and cardiovascular effects of high-dose articaine with 1:100,000 and 1:200,000 epinephrine. AB - OBJECTIVES: The authors conducted a randomized, double-blind, two-way crossover clinical trial to compare the pharmacokinetics and cardiovascular effects of 11.9 milliliters of 4 percent articaine hydrochloride (HCl) plus 1:100,000 epinephrine (A100) with those of 11.9 mL of 4 percent articaine HCl plus 1:200,000 epinephrine (A200). METHODS: During two testing sessions, the authors administered injections of A100 and A200 over a seven-minute period (in one cartridge doses unless otherwise noted): maxillary right first molar infiltration, maxillary left first molar infiltration, maxillary right first premolar infiltration, maxillary left first premolar infiltration, right inferior alveolar injection, left inferior alveolar injection, right long buccal infiltration (one-half cartridge) and left long buccal infiltration (one-half cartridge). They analyzed venous blood samples for articaine levels. They used noninvasive acoustic tonometry to measure a variety of cardiovascular parameters over a two-hour period. RESULTS: Plasma concentration curves of articaine over time were similar for both solutions, with peak concentrations and times to maximum concentration being 2,037 nanograms per milliliter and 22 minutes for A100 and 2,145 ng/mL and 22 minutes for A200. At the 10-minute point, the mean systolic blood pressure and heart rate were significantly elevated (P < .05) with A100 versus A200. CONCLUSIONS: Maximum dose recommendations for the A100 solution also can be applied to the A200 solution. A200 produces less cardiovascular stimulation than does A100. CLINICAL IMPLICATIONS: A200 is as safe as A100, and may be preferable to A100 in patients with cardiovascular disease and in those taking drugs that reportedly enhance the systemic effects of epinephrine. PMID- 17082284 TI - The anesthetic efficacy of 4 percent articaine 1:200,000 epinephrine: two controlled clinical trials. AB - OBJECTIVE: The authors conducted two double-blinded, randomized, multicenter clinical trials to determine the efficacy and clinical anesthetic characteristics of 4 percent articaine hydrochloride (HCl) with 1:200,000 epinephrine (A200) as compared with those of 4 percent articaine HCl with 1:100,000 epinephrine (A100) and 4 percent articaine HCl without epinephrine (Aw/o). METHODS: During separate testing sessions, members of the authors' research team used three articaine study formulations to induce either inferior alveolar nerve block anesthesia (Trial 1) or maxillary infiltration anesthesia (Trial 2). In each trial, subjects received, in a randomized sequence, each of the three formulations to determine efficacy (success rate) and anesthetic characteristics (onset time and duration). The authors evaluated pulpal anesthesia via subjects' response to electric pulp testing (EPT). RESULTS: A total of 126 subjects were enrolled in the two studies (63 subjects in each trial). In both mandibular and maxillary trials, the success rates for inducing profound anesthesia (EPT score > 80), the mean onset times and the mean durations of anesthesia were similar for both epinephrine-containing formulations (A200 and A100). In subjects who received the formulation containing no epinephrine (Aw/o), the success rate for profound anesthesia was significantly less. CONCLUSION: These studies demonstrated that the inclusion of epinephrine in 4 percent articaine anesthetic formulations is essential for achieving profound anesthesia. The authors found that the A200 formulation provided a level of pulpal anesthesia comparable with that of the A100 formulation. PMID- 17082285 TI - Dental caries risk in the U.S. Air Force. AB - BACKGROUND: This study describes the dental caries risk in the active duty U.S. Air Force population from October 2000 through September 2004. METHODS: The authors used data collected from two Air Force databases (personnel and dental files) by cross-referencing Social Security numbers from both databases with date. RESULTS: During the study period, the percentages of people at high and moderate risk of developing caries decreased by 31 percent and 12 percent, respectively, while the percentage of people at low risk of developing caries increased by 9 percent. Among Air Force members who were enrolled continuously during the study period, the percentages at high and moderate risk of developing caries decreased by 57 percent and 18 percent, respectively, while the percentage at low risk of developing caries increased by 14 percent. The authors observed improvement in caries risk in 83 percent and 73 percent of the people at high and moderate caries risk, respectively, for those continuously enrolled. High caries risk was related inversely to age, rank, education and years in service. Also, tobacco users had an elevated risk of developing caries. CONCLUSIONS: The Air Force Dental Service has made great strides in improving the oral health of the Air Force population. The results of this study suggest that caries risk is decreasing in the Air Force population, but oral health disparities still exist and require further evaluation. CLINICAL IMPLICATIONS: This investigation suggests that a caries risk assessment can be conducted successfully, and caries risk can be reduced by using a comprehensive population-based prevention program. This caries risk assessment also can identify factors associated with dental caries disparities in a large-scale population. PMID- 17082286 TI - High-tech dentistry: what is necessary and what is not. PMID- 17082287 TI - Is it ethical to report an insurance claim on the preparation date for a crown? PMID- 17082288 TI - For the dental patient: digital radiographs - imaging technology for the dental office. PMID- 17082291 TI - Doctors and manslaughter--response from the Crown Prosecution Service. PMID- 17082292 TI - Wet-nursing and rickets. PMID- 17082293 TI - Open access, medical research, and the internet economy of scale. PMID- 17082294 TI - Good doctors: safer patients--the Chief Medical Officer's prescription for regulating doctors. PMID- 17082295 TI - Gustave Dore's Ogre. PMID- 17082296 TI - Hubris and NEMESIS in heads of government. PMID- 17082298 TI - 'Per ardua...'Training tomorrow's surgeons using inter alia lessons from aviation. PMID- 17082299 TI - British maternal mortality in the 19th and early 20th centuries. PMID- 17082300 TI - Trench fever: the British medical response in the Great War. PMID- 17082301 TI - The World Bank's new health sector strategy: building on key assets. PMID- 17082302 TI - Mumps orchitis. AB - Although the incidence of mumps orchitis has dramatically declined since the introduction of the childhood vaccination programme, a sharp increase in reported cases of both mumps and mumps orchitis has been seen recently in the UK. There are great concerns about mumps outbreaks and the associated risk of infertility; it remains an important clinical condition. Immunization is the best policy to avoid this viral disease. PMID- 17082303 TI - Association of age, sex and deprivation with quality indicators for diabetes: population-based cross sectional survey in primary care. AB - OBJECTIVES: To determine the quality of diabetes management in primary care after the publication of the National Service Framework and examine the impact of age, gender and deprivation on the achievement of established quality indicators. DESIGN: Population-based cross sectional survey using electronic general practice records carried out between June-October 2003. SETTING: Thirty-four practices in Wandsworth, South-West London, UK. PARTICIPANTS: 6035 adult patients (> or =18 years) with diabetes from a total registered population of 201,572 patients. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Success rates for the diabetes quality indicators within the General Medical Services contract for general practitioners. RESULTS: We identified large variations in diabetes management between general practitioner practices with poorer recording of quality care in younger patients (18-44 years). In addition, younger patients had a worse cholesterol and glycaemia profile, although hypertension was more common in older patients. Gender and deprivation did not appear to be important determinants of the quality of care received. CONCLUSIONS: There are large variations in diabetes management between general practitioner practices, with care seemingly worse for younger adults. Longitudinal studies are required to determine whether current UK quality improvement initiatives have been successful in attenuating existing variations in care and treatment outcomes. PMID- 17082304 TI - Caveats in treating thyroid disease: practical implications. PMID- 17082305 TI - The first randomized trial of aspirin for heart attack and the advent of systematic overviews of trials. PMID- 17082306 TI - The big idea: iron-dependent inflammation in venous disease and proposed parallels in multiple sclerosis. PMID- 17082307 TI - Multiple pathways of selected gene amplification during adaptive mutation. AB - In a phenomenon referred to as "adaptive mutation," a population of bacterial cells with a mutation in the lac operon (lac-) accumulates Lac+ revertants during prolonged exposure to selective growth conditions (lactose). Evidence was provided that selective conditions do not increase the mutation rate but instead favor the growth of rare cells with a duplication of the leaky lac allele. A further increase in copy number (amplification) improves growth and increases the likelihood of a sequence change by adding more mutational targets to the clone (cells and lac copies per cell). These duplications and amplifications are described here. Before selection, cells with large (134-kb) lac duplications and long junction sequences (>1 kb) were common (0.2%). The same large repeats were found after selection in cells with a low-copy-number lac amplification. Surprisingly, smaller repeats (average, 34 kb) were found in high-copy-number amplifications. The small-repeat duplications form when deletions modify a preexisting large-repeat duplication. The shorter repeat size allowed higher lac amplification and better growth on lactose. Thus, selection favors a succession of gene-amplification types that make sequence changes more probable by adding targets. These findings are relevant to genetic adaptation in any biological systems in which fitness can be increased by adding gene copies (e.g., cancer and bacterial drug resistance). PMID- 17082308 TI - The protective role of silicon in the Arabidopsis-powdery mildew pathosystem. AB - The role and essentiality of silicon (Si) in plant biology have been debated for >150 years despite numerous reports describing its beneficial properties. To obtain unique insights regarding the effect of Si on plants, we performed a complete transcriptome analysis of both control and powdery mildew-stressed Arabidopsis plants, with or without Si application, using a 44K microarray. Surprisingly, the expression of all but two genes was unaffected by Si in control plants, a result contradicting reports of a possible direct effect of Si as a fertilizer. In contrast, inoculation of plants, treated or not with Si, altered the expression of a set of nearly 4,000 genes. After functional categorization, many of the up-regulated genes were defense-related, whereas a large proportion of down-regulated genes were involved in primary metabolism. Regulated defense genes included R genes, stress-related transcription factors, genes involved in signal transduction, the biosynthesis of stress hormones (SA, JA, ethylene), and the metabolism of reactive oxygen species. In inoculated plants treated with Si, the magnitude of down-regulation was attenuated by >25%, an indication of stress alleviation. Our results demonstrate that Si treatment had no effect on the metabolism of unstressed plants, suggesting a nonessential role for the element but that it has beneficial properties attributable to modulation of a more efficient response to pathogen stress. PMID- 17082309 TI - From the Academy: Colloquium review. Unique characteristics of Xanthomonas oryzae pv. oryzae AvrXa21 and implications for plant innate immunity. AB - This article provides a brief overview of some of the major concepts and molecular features of plant and animal innate immune systems. The rice pathogen recognition receptor, XA21, confers resistance to Xanthomonas oryzae pv. oryzae strains producing the AvrXa21 elicitor. Xa21 codes for a receptor-like kinase consisting of an extracellular leucine-rich repeat domain, a transmembrane domain, and a cytoplasmic kinase domain. We show that AvrXa21 activity requires the presence of rax (required for AvrXa21) A, raxB, and raxC genes that encode components of a type one secretion system. In contrast, an hrpC(-) strain deficient in type three secretion maintains AvrXa21 activity. Xanthomonas campestris pv. campestris can express AvrXa21 activity if raxST, encoding a putative sulfotransferase, and raxA are provided in trans. Expression of rax genes depends on population density and other functioning rax genes. This and other data suggest that the AvrXa21 pathogen-associated molecule is involved in quorum sensing. Together these data suggest that AvrXa21 represents a previously uncharacterized class of Gram-negative bacterial signaling molecules. These results from our studies of the XA21/AvrXa21 interaction call for some modifications in the way we think about innate immunity strategies. PMID- 17082310 TI - Dihydropyridines inhibit acetylcholine-induced hyperpolarization in cochlear artery via blockade of intermediate-conductance calcium-activated potassium channels. AB - Acetylcholine (ACh) induces hyperpolarization and dilation in a variety of blood vessels, including the cochlear spiral modiolar artery (SMA) via the endothelium derived hyperpolarization factor (EDHF). We demonstrated previously that the ACh induced hyperpolarization in the SMA originated in the endothelial cells (ECs) by activating a Ca(2+)-activated K(+) channel (K(Ca)); the hyperpolarization in smooth muscle cells was mainly an electrotonic spread via gap junction coupling. In the present study, using intracellular recording, immunohistology, and vascular diameter tracking techniques on in vitro SMA preparations, we found that 1) ACh-induced hyperpolarization was suppressed by intermediate-conductance K(Ca) (IK) blockers clotrimazole (IC(50) = 116 nM) and nitrendipine and by the calmodulin antagonist trifluoperazine, but it was not suppressed by the big conductance K(Ca) blocker iberiotoxin. The immunoreactivity to anti-SK4/IK1 antibody was localized mainly in ECs. 2) The three dihydropyridines--nifedipine, nitrendipine, and nimodipine--all concentration-dependently inhibited the ACh induced hyperpolarization, with an IC(50) value of 455, 34, and 3.2 nM, respectively. 3) Among other L-type Ca(2+) channel (I(L)) blockers, 10 microM verapamil exerted a 20% inhibition on ACh-induced hyperpolarization, whereas diltiazem and the metal ion Ca(2+) channel blockers Cd(2+) and Ni(2+) had no effect. 4) Nitrendipine and charybdotoxin abolished ACh-induced dilation in the SMA. We conclude that ACh-induced hyperpolarization in the SMA is generated mainly by activation of the IK in the ECs, and dihydropyridines suppress the EDHF mediated hyperpolarization by blocking the IK channel, not the I(L) channel. The clinical relevance of this dihydropyridine action is discussed. PMID- 17082311 TI - Modulation of synaptic transmission to second-order peripheral chemoreceptor neurons in caudal nucleus tractus solitarius by alpha1-adrenoreceptors. AB - Norepinephrine (NE) is an important neurotransmitter in central autonomic regulation. Peripheral chemoreceptor stimulation activates central noradrenergic structures. These structures innervate and therefore could modulate neurons in caudal nucleus tractus solitarius (cNTS), which receives the first central projections from peripheral chemoreceptors. However, the role of alpha(1) adrenoreceptors in synaptic transmission of peripheral chemoreceptor inputs in cNTS is unknown. We investigated the responses to activation of alpha(1) adrenoreceptors on glutamatergic and GABAergic inputs in NTS slices using whole cell recording. Second-order neurons were identified by 1,1'-dilinoleyl-3,3,3',3' tetra-methylindocarbocyanine, 4-chlorobenzenesulphonate (DiA) labeling of carotid bodies. Electrical stimulation of ipsilateral tractus solitarius was used to evoke excitatory postsynaptic currents (eEPSCs), whereas inhibitory postsynaptic currents were evoked (eIPSCs) by electrically stimulating NTS near the recorded neuron. Application of alpha(1)-adrenoreceptor agonist phenylephrine (PE) at 20 microM significantly decreased amplitudes of eEPSCs (78 +/- 1% of control; n = 16; p < 0.01), and it increased amplitudes of eIPSCs (120 +/- 13% of control; n = 7; p < 0.01). Both effects were blocked by the alpha(1)-adrenoreceptor antagonist prazosin at 10 microM. PE did not change holding current, input resistance, and current-voltage relationship in cNTS neurons. PE significantly changed paired pulse ratios of eEPSC/eIPSCs, increased the frequency of miniature IPSCs (329 +/- 10% of control; n = 6; p < 0.05), but it decreased that of miniature EPSCs (69 +/ 6% of control; n = 5; p < 0.01). PE-induced inhibition of eEPSCs was independent of N-methyl-D-aspartate or GABA(B) receptors. These results suggest that activation of alpha(1)-adrenoreceptors reduces excitatory and enhances inhibitory inputs to second-order peripheral chemoreceptor neurons in cNTS via a presynaptic mechanism. These actions result in the inhibition of synaptic transmission and could play a role in the autonomic responses to hypoxia. PMID- 17082312 TI - Leptin antagonist reveals an uncoupling between leptin receptor signal transducer and activator of transcription 3 signaling and metabolic responses with central leptin resistance. AB - Leptin-resistant rats have reduced leptin receptors and signaling and are refractory to exogenous leptin. However, it is unclear how leptin-mediated hypothalamic signal transducer and activator of transcription 3 (STAT3) signaling relates to the loss of physiological responsiveness. We hypothesized that if leptin resistance is associated with leptin receptors that are no longer functionally coupled to leptin responses, then a leptin antagonist should be less effective in leptin-resistant compared with leptin-responsive rats. Hypothalamic leptin resistance was induced in lean rats with a recombinant adeno-associated viral (rAAV) vector encoding leptin by intracerebroventricular injection. Following development of leptin resistance, at day 153, these rats and control rats were infused centrally either with vehicle or a rat leptin antagonist for 14 days. Food intake, body weight, adiposity, and uncoupling protein 1 expression increased with antagonist infusion in controls but elevated only marginally in leptin-resistant rats. Basal hypothalamic STAT3 signaling remained unchanged with antagonist infusion in control rats despite the pronounced orexigenic response in these animals. STAT3 phosphorylation in rats pretreated with rAAV-leptin to induce leptin resistance was elevated 2-fold. Paradoxically, in these leptin resistant rats, the antagonist fully reversed the 2-fold elevated phosphorylated STAT3, but it evoked minimal physiological responses. These data reveal an uncoupling between leptin receptor activation and metabolic responses with central leptin resistance. PMID- 17082313 TI - 17beta-estradiol antagonizes the down-regulation of endothelial nitric-oxide synthase and GTP cyclohydrolase I by high glucose: relevance to postmenopausal diabetic cardiovascular disease. AB - In postmenopausal women, the risk of diabetic cardiovascular disease drastically increases compared with that of men or premenopausal women. However, the mechanism of this phenomenon has not yet been clarified. We hypothesized that the beneficial effects of estrogen on endothelial function may be relevant to protection against hyperglycemia-induced vascular derangement. Bovine aortic endothelial cells were incubated for 72 h in the presence and absence of the physiological concentration of 17beta-estradiol (17beta-E2) under normal and high glucose conditions. The presence of 17beta-E2 significantly counteracted the reduction in basal nitric oxide production under high-glucose conditions. This finding was associated with the recovery of endothelial nitric-oxide synthase (eNOS) protein expression, tetrahydrobiopterin (BH4) levels, and the activity and gene expression of GTP cyclohydrolase I (GTPCH-I), a rate-limiting enzyme for BH4 synthesis. Both the gene transfer of estrogen receptor alpha using adenovirus and treatment with the protein kinase C inhibitor bisindolylmaleimide I significantly enhanced the effects of 17beta-E2 treatment under high-glucose conditions, whereas these effects were abolished by the estrogen receptor antagonist ICI 182,780 (faslodex). Transfection of small-interfering RNA targeting eNOS resulted in a marked reduction in GTPCH-I mRNA under both normal and high-glucose conditions, but this reduction was strongly reversed by 17beta-E2. These results suggest that the activation of ERalpha with 17beta-E2 can counteract high-glucose induced down-regulation of eNOS and GTPCH-I in endothelial cells. Therefore, estrogen deficiency may result in an exaggeration of hyperglycemia-induced endothelial dysfunction, leading to the development of cardiovascular disease in postmenopausal diabetic women. PMID- 17082314 TI - Progressive immunoglobulin gene mutations in chronic lymphocytic leukemia: evidence for antigen-driven intraclonal diversification. AB - Somatic mutations of immunoglobulin genes characterize mature memory B cells, and intraclonal B-cell diversification is typically associated with expansion of B cell clones with greater affinity for antigen (antigen drive). Evidence for a role of antigen in progression of intraclonal chronic lymphocytic leukemia (CLL) cell diversification in patients with mutated immunoglobulin genes has not been previously presented. We performed a single-cell analysis of immunoglobulin heavy and light chains in 6 patients with somatically mutated CLL-cell immunoglobulin genes and identified 2 patients with multiple related (oligoclonal) subgroups of CLL cells. We constructed genealogic trees of these oligoclonal CLL-cell subgroups and assessed the effects of immunoglobulin somatic mutations on the ratios of replacement and silent amino acid changes in the framework and antigen binding regions (CDRs) of the immunoglobulin heavy and light chains from each oligoclonal CLL-cell population. In one subject, the amino acid changes were consistent with an antigen-driven progression of clonally related CLL-cell populations. In the other subject, intraclonal diversification was associated with immunoglobulin amino acid changes that would have likely lessened antigen affinity. Taken together, these studies support the hypothesis that in some CLL cases intraclonal diversification is dependent on antigen interactions with immunoglobulin receptors. PMID- 17082315 TI - Pathologic consequences of STAT3 hyperactivation by IL-6 and IL-11 during hematopoiesis and lymphopoiesis. AB - We have previously demonstrated that STAT3 hyperactivation via the interleukin 6 (IL-6) cytokine family receptor gp130 in gp130 (Y757F/Y757F) mice leads to numerous hematopoietic and lymphoid pathologies, including neutrophilia, thrombocytosis, splenomegaly, and lymphadenopathy. Because IL-6 and IL-11 both signal via a gp130 homodimer, we report here a genetic approach to dissect their individual roles in these pathologies. Neutrophilia and thrombocytosis were absent in gp130 (Y757F/Y757F) mice lacking either IL-6 (gp130 (Y757F/Y757F): IL-6 (-/-)) or the IL-11 receptor alpha subunit (gp130 (Y757F/Y757F): IL-11Ralpha1 (-/ )), and this was associated with a normalized bone marrow compartment. The elevated myelopoiesis and megakaryopoiesis in bone marrow of gp130 (Y757F/Y757F) mice was attributable to an increase by either IL-6 or IL-11 in the STAT3-driven impairment of transforming growth factor beta (TGF-beta) signaling, which is a suppressor of these lineages. In contrast, the absence of IL-6, but not IL-11 signaling, prevented the splenomegaly, abnormal lymphopoiesis, and STAT3 hyperactivation in lymphoid organs of gp130 (Y757F/Y757F) mice. Furthermore, hyperactivation of STAT3 in lymphoid organs was associated with increased expression of IL-6Ralpha, and IL-6Ralpha expression was reduced in gp130 (Y757F/Y757F): Stat3 (+/-) mice displaying normal levels of STAT3 activity. Collectively, these data genetically define distinct roles of IL-6 and IL-11 in driving pathologic hematopoietic and lymphoid responses mediated by STAT3 hyperactivation. PMID- 17082316 TI - Facilitating matched pairing and expression of TCR chains introduced into human T cells. AB - Adoptive transfer of T lymphocytes is a promising treatment for a variety of malignancies but often not feasible due to difficulties generating T cells that are reactive with the targeted antigen from patients. To facilitate rapid generation of cells for therapy, T cells can be programmed with genes encoding the alpha and beta chains of an antigen-specific T-cell receptor (TCR). However, such exogenous alpha and beta chains can potentially assemble as pairs not only with each other but also with endogenous TCR alpha and beta chains, thereby generating alphabetaTCR pairs of unknown specificity as well as reducing the number of exogenous matched alphabetaTCR pairs at the cell surface. We demonstrate that introducing cysteines into the constant region of the alpha and beta chains can promote preferential pairing with each other, increase total surface expression of the introduced TCR chains, and reduce mismatching with endogenous TCR chains. This approach should improve both the efficacy and safety of ongoing efforts to use TCR transfer as a strategy to generate tumor-reactive T cells. PMID- 17082317 TI - Respiratory tract infections and subsequent risk of chronic lymphocytic leukemia. AB - Recent evidence suggests that chronic lymphocytic leukemia (CLL) might occur following a response to an infectious agent. We conducted a population-based study including 4249 CLL patients diagnosed in Denmark from 1977 to 1997 and 15 690 frequency-matched controls to quantify risk of CLL following various airway infections. Through data linkage we gathered information on hospital inpatient/outpatient discharges that listed infections present at least 1 year prior to CLL. Using logistic regression, we calculated odds ratios (ORs) and 95% confidence intervals (CIs). Personal history of pneumonia was associated with significantly increased CLL risk (OR = 1.4; 1.2-1.8); risk was restricted to 1 to 4.99 years prior to CLL diagnosis (OR = 1.6; 1.2-2.0). Individuals with 3 or more prior pneumonia events had a significant 2.5-fold (1.1-5.6) elevated CLL risk, and risk increased with the number of pneumonia episodes (P(trend) < .001). None of 9 other respiratory-tract infections was significantly associated with CLL risk. Pneumonia might be a potential CLL trigger or it could represent premalignant immune disruption preceding CLL. PMID- 17082318 TI - Human plasma fibrinogen is synthesized in the liver. AB - Hereditary systemic amyloidosis caused by fibrinogen Aalpha-chain gene mutations is an autosomal dominant condition with variable penetrance, usually of late onset, and typically presents with nephropathy leading to renal failure. Amyloid deposits often develop rapidly in transplanted kidneys, and concomitant orthotopic liver transplantation has lately been performed in several patients with the hope of halting amyloid deposition. Fibrinogen is produced in vitro by hepatocytes but also by other human cell types, and although the liver is the source of plasma fibrinogen in vivo in rats, this is not known in humans. Transplantation of livers expressing wild-type fibrinogen into patients with variant fibrinogen amyloidosis provides a unique opportunity to establish the source of human plasma fibrinogen. We therefore characterized plasma fibrinogen Aalpha-chain allotypes by electrospray ionization mass spectrometry mapping of tryptic digests before and after liver transplantation. Before liver transplantation, fibrinogen amyloidosis patients with the Glu526Val Aalpha-chain variant had approximately equal proportions of peptide with the wild-type sequence TFPGFFSPMLGEFVSETESR, and with the amyloidogenic variant sequence TFPGFFSPMLGEFVSVTESR, as expected for individuals heterozygous for the mutation. After transplantation, only the wild-type sequence was detected, and the liver is thus the source of at least 98% of the circulation fibrinogen. PMID- 17082319 TI - Thrombospondin-1 limits ischemic tissue survival by inhibiting nitric oxide mediated vascular smooth muscle relaxation. AB - The nitric oxide (NO)/cGMP pathway, by relaxing vascular smooth muscle cells, is a major physiologic regulator of tissue perfusion. We now identify thrombospondin 1 as a potent antagonist of NO for regulating F-actin assembly and myosin light chain phosphorylation in vascular smooth muscle cells. Thrombospondin-1 prevents NO-mediated relaxation of precontracted vascular smooth muscle cells in a collagen matrix. Functional magnetic resonance imaging demonstrated that an NO mediated increase in skeletal muscle perfusion was enhanced in thrombospondin-1 null relative to wild-type mice, implicating endogenous thrombospondin-1 as a physiologic antagonist of NO-mediated vasodilation. Using a random myocutaneous flap model for ischemic injury, tissue survival was significantly enhanced in thrombospondin-1-null mice. Improved flap survival correlated with increased recovery of oxygen levels in the ischemic tissue of thrombospondin-1-null mice as measured by electron paramagnetic resonance oximetry. These findings demonstrate an important antagonistic relation between NO/cGMP signaling and thrombospondin-1 in vascular smooth muscle cells to regulate vascular tone and tissue perfusion. PMID- 17082320 TI - A GMCSF and IL-15 fusokine leads to paradoxical immunosuppression in vivo via asymmetrical JAK/STAT signaling through the IL-15 receptor complex. AB - We hypothesized that a granulocyte macrophage colony-stimulating factor (GMCSF) and interleukin 15 (IL-15) fusokine (GIFT15) would possess greater immune stimulatory properties than their combined use. Unexpectedly, tumor cells engineered to secrete GIFT15 protein led to suppression of natural killer (NK) and NKT-cell recruitment in vivo, suggesting an unanticipated immune-suppressive effect. We found GIFT15 to have pleiotropic effects on an array of immune competent cells. Among these, macrophages treated with GIFT15 secrete de novo the tissue inhibitor of metalloproteinase-2 (TIMP-2); activated matrix metalloproteinase-2 (MMP-2); transforming growth factor-beta (TGF-beta); as well as vascular endothelial growth factor (VEGF). We show that the GIFT15 fusokine has increased affinity for the alpha chain component of the IL-15R, leading to aberrant signaling through the beta chain manifested by the hyperphosphorylation of STAT3 both in macrophages and splenocytes. Suppression of common gamma chain mediated STAT5 phosphorylation and blockade of the IL-15-dependent IFN-gamma response in mouse splenocytes were also observed. We tested GIFT15 as an immunosuppressor and demonstrated that it allowed engraftment of allogeneic B16F0 and human xenograft U87GM glioma cells in immunocompetent mice. Thus, GIFT15 defines a new class of fusokine that mediates proangiogenic and immunosuppressive effects via aberrant signaling by the IL-15R in lymphomyeloid cells. PMID- 17082321 TI - Active tissue factor pathway inhibitor is expressed on the surface of coated platelets. AB - The incorporation of blood-borne forms of tissue factor (TF) into a growing blood clot is necessary for normal fibrin generation and stabilization of the blood clot. Tissue factor pathway inhibitor (TFPI) is the primary physiologic inhibitor of tissue factor and is present within platelets. Expression of TFPI on the platelet surface may be the optimal location for it to abrogate blood-borne TF activity that incorporates within the blood clot, balancing the need for adequate hemostasis while preventing development of occlusive thrombosis. TFPI is produced by megakaryocytes but is not expressed on the platelet surface. Activation of platelets with thrombin receptor activation peptide does not cause release or surface expression of TFPI, demonstrating that TFPI is not stored within platelet alpha granules. TFPI is expressed on the platelet surface following dual-agonist activation with convulxin plus thrombin to produce coated platelets. In association with its expression on the surface of coated platelets TFPI is also released in microvesicles or as a soluble protein. PMID- 17082322 TI - Rapamycin is efficacious against primary effusion lymphoma (PEL) cell lines in vivo by inhibiting autocrine signaling. AB - The antitumor potency of the mTOR inhibitor rapamycin (sirolimus) is the subject of intense investigations. Primary effusion lymphoma (PEL) appears as an AIDS defining lymphoma and like Kaposi sarcoma has been linked to Kaposi sarcoma associated herpesvirus (KSHV). We find that (1) rapamycin is efficacious against PEL in culture and in a murine xenograft model; (2) mTOR, its activator Akt, and its target p70S6 kinase are phosphorylated in PEL; (3) rapamycin inhibits mTOR signaling as determined by S6 phosphorylation; (4) KSHV transcription is unaffected; (5) inhibition of IL-10 signaling correlates with drug sensitivity; and (6) addition of exogenous IL-10 or IL-6 can reverse the rapamycin growth arrest. This validates sirolimus as a new treatment option for PEL. PMID- 17082323 TI - A phase 2 study of the farnesyltransferase inhibitor tipifarnib in poor-risk and elderly patients with previously untreated acute myelogenous leukemia. AB - Outcomes for older adults with acute myelogenous leukemia (AML) are poor due to both disease and host-related factors. In this phase 2 study, we tested the oral farnesyltransferase inhibitor tipifarnib in 158 older adults with previously untreated, poor-risk AML. The median age was 74 years, and a majority of patients had antecedent myelodysplastic syndrome. Complete remission (CR) was achieved in 22 patients (14%); partial remission or hematologic improvement occurred in 15 patients, for an overall response rate of 23%. The median duration of CR was 7.3 months and the median survival of complete responders was 18 months. Adverse karyotype, age 75 years or older, and poor performance status correlated negatively with survival. Early death in the absence of progressive disease was rare, and drug-related nonhematologic serious adverse events were observed in 74 patients (47%). Inhibition of farnesylation of the surrogate protein HDJ-2 occurred in the large majority of marrow samples tested. Baseline levels of phosphorylated mitogen-activated protein kinase and AKT did not correlate with clinical response. Tipifarnib is active and well tolerated in older adults with poor-risk AML and may impart a survival advantage in those patients who experience a clinical response. PMID- 17082324 TI - The role of TLR2 in the inflammatory activation of mouse fibroblasts by human antiphospholipid antibodies. AB - Antiphospholipid antibodies (APLAs) promote inflammatory and procoagulant responses in endothelial cells and monocytes. Previous studies have shown that MyD88, TRAF6, and NF-kappaB mediate cell activation by APLAs. These intermediates are also used by toll-like receptors (TLRs). We investigated the role of TLRs in the cellular response to APLAs. IgGs were isolated from the plasma of 5 patients with antiphospholipid syndrome along with immunopurified anti-beta2-glycoprotein 1 IgG from a sixth patient. Control IgG was obtained from a pool of healthy donor plasmas negative for APLAs. Wild-type mouse embryonic fibroblasts (EFs) and EFs deficient in TLR1, TLR2, TLR4, or TLR6 were incubated with APLAs, anti-beta2 glycoprotein 1 IgG, or control IgG. On incubation with the patient IgG, but not control IgG, a significant increase in mRNA levels of the inflammatory marker proteins MCP-1, ICAM-1, and IL-6 as well as IL-6 secretion was observed in wild type EFs, whereas TLR2-deficient EFs did not respond. Responses in TLR1- and TLR6 deficient EFs were decreased and those in TLR4-deficient EFs comparable to those in wild-type EFs. Overexpression of human TLR2 in the TLR2-deficient EFs restituted the response to patient IgG. Our results imply that TLR2 plays a role in mouse fibroblast activation by APLAs. PMID- 17082325 TI - Pituitary-interrenal interaction in zebrafish interrenal organ development. AB - To further elucidate pituitary adrenal interactions during development, we studied the organogenesis of the interrenal organ, the teleost homolog of the mammalian adrenal gland, in zebrafish. To this end we compared wild-type zebrafish interrenal development with that of mutants lacking pituitary cell types including corticotrophs. In addition, we studied the effects of ACTH receptor (Mc2r) knockdown and dexamethasone (dex) on interrenal development and pituitary feedback. Until 2 d post fertilization (2 dpf) interrenal development assessed by transcripts of key steroidogenic genes (cyp11a1, mc2r, star) is independent of proopiomelanocortin (Pomc) as demonstrated in aal/eya1and lia/fgf3 mutants. However, at 5 dpf lack of pituitary cells leads to reduced expression of steroidogenic genes at both the transcriptional and the protein level. Pituitary control of interrenal development resides in corticotrophs, because pit1 mutants lacking pituitary cells except corticotrophs have a phenotype similar to that of wild-type controls. Furthermore, development in mc2r knockdown morphants does not differ from aal/eya1 and lia/fgf3 mutants. Inhibition of steroidogenesis by mc2r knockdown induces up-regulation of pomc expression in the anterior domain of pituitary corticotrophs. Accordingly, dex suppresses pomc in the anterior domain only, leading to impaired expression of steroidogenic genes commencing at 3 dpf and interrenal hypoplasia via reduced interrenal proliferation. In contrast, negative feedback on pituitary corticotrophs by dex is evident at 2 dpf and precedes effects of Pomc on the interrenal primordium. These data demonstrate a gradual transition from early pituitary-independent interrenal organogenesis to developmental control by the anterior domain of pituitary corticotrophs acting via Mc2 receptors. PMID- 17082326 TI - Mapping the binding site of arginine vasopressin to V1a and V1b vasopressin receptors. AB - Starting from the 2.8-A resolution x-ray structure of bovine rhodopsin, three dimensional molecular models of the complexes between arginine vasopressin and two receptor subtypes (V1a, V1b) have been built. Amino acid sequence alignment and docking studies suggest that four key residues (1.35, 2.65, 4.61, and 5.35) fine tune the binding of vasopressin and related peptide agonists to both receptor subtypes. To validate these predictions, a series of single or double mutants were engineered at V1a and V1b receptor subtypes and tested for their binding and functional properties. Two negatively charged amino acids at positions 1.35 and 2.65 are key anchoring residues to the Arg8 residue of arginine vasopressin. Moreover, two amino acids (V(4.61) and P(5.35)) delineating a hydrophobic subsite at the human V1b receptor are responsible for the recognition of V1b selective peptide agonists. Last, one of the latter positions (5.35) is hypothesized to explain the pharmacological species differences between rat and human vasopressin receptors for a V1b peptide agonist. Altogether these refined three-dimensional models of V1a and V1b human receptors should enable the identification of further new selective V1a and V1b agonists as pharmacological but also therapeutic tools. PMID- 17082327 TI - Transgelin functions as a suppressor via inhibition of ARA54-enhanced androgen receptor transactivation and prostate cancer cell growth. AB - The androgen receptor (AR) requires coregulators for its optimal function. However, whether AR coregulators further need interacting protein(s) for their proper function remains unclear. Here we describe transgelin as the first ARA54 associated negative modulator for AR. Transgelin suppressed ARA54-enhanced AR function in ARA54-positive, but not in ARA54-negative, cells. Transgelin suppressed AR transactivation via interruption of ARA54 homodimerization and AR ARA54 heterodimerization, resulting in the cytoplasmic retention of AR and ARA54. Stable transfection of transgelin in LNCaP cells suppressed AR-mediated cell growth and prostate-specific antigen expression, whereas this suppressive effect was abolished by the addition of ARA54-small interfering RNA. Results from tissue surveys showing decreased expression of transgelin in prostate cancer specimens further strengthened the suppressor role of transgelin. Our findings reveal the novel mechanisms of how transgelin functions as a suppressor to inhibit prostate cancer cell growth. They also demonstrate that AR coregulators, like ARA54, might have dual in vivo roles functioning as both a direct coactivator and as an indirect mediator in AR function. The finding that a protein can modulate AR function without direct interaction with AR might provide a new therapeutic approach, with fewer side effects, to battle prostate cancer by targeting AR indirectly. PMID- 17082328 TI - Technical factors in fractal analysis of periapical radiographs. AB - OBJECTIVES: Fractal analysis quantifies complex geometric structures by generating a fractal dimension, which can measure trabecular bone density. The use of non-standardized radiographic techniques potentially limits the reliability of fractal analysis. The objective of this study was to determine how variations in radiographic technique affect fractal dimension. METHODS: Periapical radiographs of maxillary incisors taken on eight dry human skulls at varying angulation, tube potential and impulse settings were subjected to Fourier transform fractal analysis. RESULTS: A significant (p < 0.05) difference in fractal dimension, ranging from 2.5 to 2.9, was found between the skulls, potentially reflecting variations in bone trabeculation. However, variations in angulation, tube potential or impulse did not have a statistically significant effect (p > 0.05) on fractal dimension, with the standard deviation of the fractal dimension ranging from +/- 0.005 to +/- 0.062 at various radiographic machine settings. CONCLUSIONS: This study indicates that non-standardized periapical radiographs may provide a reliable method for determining fractal dimensions, which could be useful in analysing changes in alveolar bone density in various bone disorders. PMID- 17082329 TI - Development of a cone angle weighted three-dimensional image reconstruction algorithm to reduce cone-beam artefacts. AB - OBJECTIVES: Image reconstruction from cone-beam projections collected along a single circular source trajectory is commonly done using the Feldkamp algorithm, which performs well only with a small cone angle. In this report, we propose an algorithm to reduce cone-beam artefacts by increasing the cone angle by several fold to achieve satisfactory image quality at the same radiation dose. METHODS: To examine the factors involved in the occurrence of cone-beam artefacts, a microspheres-phantom was arranged longitudinally at different positions and a computer simulation was performed. Due to differences in projection angle, data projected onto the detector surface were projected along trajectories shown as different periodic functions depending on the distance and position from the mid plane position. Therefore, projection along several detector channels based on different projection data resulting from different periodic functions is considered responsible for the increase in cone-beam artefacts associated with an increase in the distance of reconstruction planes from the mid-plane position. Our recommended algorithm to reduce such artefacts features a change in weighting with respect to projection data obtained at different projection angles, three dimensional back-projection of corrected projection data. RESULTS: Numerical phantom simulation and real human head origin study (a prototype cone-beam CT) showed that the effect of the reduction in cone-beam artefacts of an object located at the edges was markedly enhanced at reconstruction planes at positions further from the mid-plane position. CONCLUSION: We propose a projection angle weight-based algorithm to increase the cone angle by several fold to achieve satisfactory image quality at the same radiation dose. These findings confirmed that this algorithm reduces cone-beam artefacts and generates high-quality reconstruction images. PMID- 17082330 TI - Density conversion factor determined using a cone-beam computed tomography unit NewTom QR-DVT 9000. AB - OBJECTIVE: The purpose of this study was to determine a conversion coefficient for Hounsfield Units (HU) to material density (g cm(-3)) obtained from cone-beam computed tomography (CBCT-NewTom QR-DVT 9000) data. METHODS: Six cylindrical models of materials with different densities were made and scanned using the NewTom QR-DVT 9000 Volume Scanner. The raw data were converted into DICOM format and analysed using Merge eFilm and AMIRA to determine the HU of different areas of the models. RESULTS: There was no significant difference (P = 0.846) between the HU given by each piece of software. A linear regression was performed using the density, rho (g cm(-3)), as the dependent variable in terms of the HU (H). The regression equation obtained was rho = 0.002H-0.381 with an R2 value of 0.986. The standard error of the estimation is 27.104 HU in the case of the Hounsfield Units and 0.064 g cm(-3) in the case of density. CONCLUSION: CBCT provides an effective option for determination of material density expressed as Hounsfield Units. PMID- 17082331 TI - Accuracy of three-dimensional measurements using cone-beam CT. AB - OBJECTIVES: Lesions causing intraosseous defects in the head and neck region are difficult to diagnose using two-dimensional radiography, and three-dimensional (3D) data provided by CT is useful but often difficult to obtain. Recently, cone beam CT (CBCT) was made available, with the potential to become a practical tool in dentistry. However, there is limited evidence to prove that defect volume can be determined accurately. Therefore, this in vitro validation study aimed at establishing whether linear and 3D CBCT, using volumetric measurements, is accurate for determining osseous defect sizes. METHODS: Depth and diameter of simulated bone defects in (i) an acrylic block and (ii) a human mandible were blindly measured electronically by five examiners using CBCT. Linear measurements were compared with predetermined machined dimensions. Using software, volume extraction was performed by another examiner on the acrylic phantom and compared with known dimensions. Data were analysed using paired t-tests. RESULTS: Using the acrylic block, mean width accuracy was -0.01 mm (+/- 0.02 SE) and mean height difference was -0.03 mm (+/- 0.01 SE; P > 0.05). For the human mandible, mean width accuracy was -0.07 mm (+/- 0.02 SE) and mean height accuracy was -0.27 mm (+/- 0.02 SE; P < 0.01). Volume accuracy was -6.9 mm3 (+/- 4 SE) for automated calculations and -2.3 mm3 (+/- 2.6 SE) for the manual measurements (P < 0.001). CONCLUSIONS: CBCT has the potential to be an accurate, non-invasive, practical method to reliably determine osseous lesion size and volume. Further clinical validation will lead to a vast array of applications in oral and maxillofacial diagnosis. PMID- 17082332 TI - Clinical efficacy of a new software developed for dental digital subtraction radiography. AB - OBJECTIVES: The aim of this study was to test and compare the efficacy of software developed recently for digital subtraction radiography (DSR) in vivo. METHODS: An algorithm performing both manual and automated image reconstructions and contrast correction was developed for the manipulation of radiographic images. Pre- and post-operative radiographic images of ten patients were obtained and the automated subtraction analyses were performed using four different softwares (new software, Emago, Photoshop 8.0 and Paintshop Pro 9). Ten experienced dental specialists evaluated the clinical efficacy of each program and scored the softwares by using visual analogue scales (VAS). The results were statistically analysed and alpha was set as 0.05. RESULTS: The newly developed algorithm received higher scores than the others (new software =67.89, Emago = 64.26, Paintshop Pro 9 = 33.41 and Photoshop 8.0 = 27.24, respectively). The clinical efficacies of the new software and Emago were not significantly different (P = 0.720); likewise, Photoshop 8.0 and Paintshop Pro 9 performed comparably (P = 0.295). CONCLUSIONS: Considering this study, the new software and Emago would be suggested for DSR in dental practice. PMID- 17082333 TI - Comparison of simulated periodontal bone defect depth measured in digital radiographs in dedicated and non-dedicated software systems. AB - OBJECTIVES: To compare simulated periodontal bone defect depth measured in digital radiographs with dedicated and non-dedicated software systems and to compare the depth measurements from each program with the measurements in dry mandibles. METHODS: Forty periodontal bone defects were created at the proximal area of the first premolar in dry pig mandibles. Measurements of the defects were performed with a periodontal probe in the dry mandible. Periapical digital radiographs of the defects were recorded using the Schick sensor in a standardized exposure setting. All images were read using a Schick dedicated software system (CDR DICOM for Windows v.3.5), and three commonly available non dedicated software systems (Vix Win 2000 v.1.2; Adobe Photoshop 7.0 and Image Tool 3.0). The defects were measured three times in each image and a consensus was reached among three examiners using the four software systems. The difference between the radiographic measurements was analysed using analysis of variance (ANOVA) and by comparing the measurements from each software system with the dry mandibles measurements using Student's t-test. RESULTS: The mean values of the bone defects measured in the radiographs were 5.07 mm, 5.06 mm, 5.01 mm and 5.11 mm for CDR Digital Image and Communication in Medicine (DICOM) for Windows, Vix Win, Adobe Photoshop, and Image Tool, respectively, and 6.67 mm for the dry mandible. The means of the measurements performed in the four software systems were not significantly different, ANOVA (P = 0.958). A significant underestimation of defect depth was obtained when we compared the mean depths from each software system with the dry mandible measurements (t-test; P approximately equal to 0.000). CONCLUSIONS: The periodontal bone defect measurements in dedicated and in three non-dedicated software systems were not significantly different, but they all underestimated the measurements when compared with the measurements obtained in the dry mandibles. PMID- 17082334 TI - The influence of the X-ray spectrum at compact bone-titanium interfaces in digital dental radiography. AB - OBJECTIVES: To determine the optimum X-ray spectrum in digital dental radiography once the dose around an implant and the diagnostic usefulness of the image are taken into account. MATERIALS AND METHODS: A Monte Carlo code (MCNP4B) was employed for computing the dose distribution across the bone-titanium interface. The X-ray spectra used were those met in digital dental radiography; 50-70 kVp, 2 mm Al total filtration, 5 kVp increment. RESULTS: The variation of the ratio of dose with as opposed to without implant against depth reaches maximum values at the bone-implant interface that vary between 2.9 and 3.2. For the same number of photon histories followed, the higher the tube potential setting, the greater the dose both in contact and inside the implant. CONCLUSION: In digital dental radiography, a 60-65 kVp spectrum accompanied by the known 30% reduction in mAs leads to lower dose to the patient for a diagnostically useful image. PMID- 17082335 TI - Cytokine profile in synovial fluid from patients with internal derangement of the temporomandibular joint: a preliminary study. AB - OBJECTIVES: Temporomandibular joint disorders (TMD) comprise a group of chronic painful conditions of mastication in the temporomandibular joint (TMJ). Although the association between TMD and internal derangement of the TMJ is well documented, the functional relevance is still unclear. Increased concentrations of inflammatory mediators have been identified in the synovial fluid of affected patients with TMD, suggesting an underlying degenerative or inflammatory process. The aim of this study was to generate a comprehensive cytokine expression profile in TMD. METHODS: 15 samples from patients with internal derangement of TMJ were analysed using a novel cytokine array that enables the analysis of 79 different cytokines simultaneously. RESULTS: Cytokine levels were correlated with the presence of joint effusion (JE) determined by MRI. In the majority of synovial fluid samples, angiogenin (Ang), fibroblast growth factor (FGF)-9, insulin-like growth factor-binding protein (IGFBP)-3, interleukin (IL)-1alpha, IL-1beta, IL-8, inducible protein (IP)-10, macrophage inflammatory protein (MIP)-1beta, osteoprotegerin (OPG), transforming growth factor (TGF)-beta2, tissue inhibitor of metalloproteinase (TIMP)-1, TIMP-2, tumour necrosis factor (TNF)-beta and vascular endothelial growth factor (VEGF) were detectable. Furthermore, the expression levels of Ang, brain-derived neurotrophic factor (BDNF), FGF-4, FGF-9, IGFBP-2, IL-8, MIP-1beta, OPG, pulmonary and activation-regulated protein (PARC), TGF-beta2, TIMP-2 and VEGF were significantly associated with the presence of JE; among these, nine cytokines (Ang, BDNF, FGF-4, FGF-9, IGFBP-2, MIP-1beta, PARC, TGF-beta2 and TIMP-2) were hitherto not described in TMD. CONCLUSIONS: This study confirmed previous reports of elevated cytokine levels in TMD. Additionally, we identified previously undescribed cytokines that were upregulated and correlated significantly with the presence of JE. We were able to identify novel cytokines that have hitherto not been described in TMD. Strategies targeting the identified cytokines may represent a novel therapy option in TMD. PMID- 17082336 TI - The precision of the panoramic mandibular index. AB - OBJECTIVES: This study evaluates the precision of the panoramic mandibular index (PMI). MATERIALS AND METHODS: Measurements were made by two observers on both of the left and right sides of the mandible on the radiographs taken from 41 young individuals and the inferior and superior PMI were calculated. Two weeks later the same observers repeated their measurements to assess intrarater reliability. Paired t-test, Pearson correlation coefficients and precision values were calculated to assess levels of association. RESULTS: There was a significant difference between the first rater's first and second measurements (intrarater repeatability) for both the inferior and superior PMI, while no significant difference was observed for the second rater. The intrarater and inter-rater precision values for the inferior PMI were calculated as 0.005319 and 0.005594 for the first rater and 0.005663 and 0.005594 for the second, respectively. The intrarater and inter-rater precision values for the superior PMI were similar for both observers, calculated as 0.002558, 0.002766; and 0.003046, 0.002766, respectively. The precision of both inferior and superior indices was not very good, but precision figures for the superior PMI demonstrated consistency almost twice better than those for the inferior PMI for both observers. CONCLUSIONS: The precision values for the PMI seem to be sufficient according to the results of this study, but they are still questionable and more studies need to be done on this aspect. PMID- 17082337 TI - A case of frontal lobe abscess as a complication of frontal sinus ossifying fibroma. AB - Sinonasal ossifying fibroma is a rare, slow-growing, benign bony tumour, frequently involving the maxilla and mandible in the head and neck region. Although it is known to be the second most frequent fibro-osseous tumour of paranasal sinus, to the best of our knowledge, ossifying fibroma of frontal sinus causing brain abscess has not been presented yet in the relevant literature. We present the clinical, pathological and radiological findings of ossifying fibroma of the frontal sinus associated with brain abscess. PMID- 17082338 TI - Embolisation of arteriovenous malformation of the maxilla. AB - We describe a 20-year-old male patient who presented with gingival bleeding. Physical examination showed gingival swelling of the right maxilla and loosening of the molar teeth. The initial diagnosis of gingivitis was made, but further examination revealed a lytic lesion of the maxilla. On suspicion of fibrous dysplasia, biopsy was attempted but was unsuccessful due to severe haemorrhage. Further evaluation showed palpable and audible bruit on the gingiva, which caused the suspicion of vascular malformation. Angiography was performed and demonstrated arteriovenous malformation (AVM). Embolisation therapy with polyvinyl alcohol was performed. Post-embolisation angiogram demonstrated complete obliteration of the lesion. PMID- 17082339 TI - Schwannoma with secondary erosion of mandible: case report with a review of the literature. AB - Schwannoma (neurilemmoma) is a common, histologically distinctive, benign, usually encapsulated, peripheral nerve tumour of Schwann cell origin. We report a case of schwannoma arising from soft tissue near the mandible. A 53-year-old female presented at our department with painless swelling of lingual mucosa of the mandible. The first molar was extracted 20 months before. Panoramic radiograph showed a suspected residual cyst. It was impossible to determine prior to surgery whether this was a peripheral nerve sheath tumour. The lesion was completely removed; the tumour appears to have originated in soft tissue and caused secondary erosion of the mandible. PMID- 17082340 TI - Bilateral mandibular condylysis in a patient with systemic sclerosis. AB - A case of progressive osteolysis of the mandibular condyles in a 55-year-old female patient with systemic sclerosis (SSc) is described. Orofacial involvement and radiological manifestations of SSc are discussed. PMID- 17082341 TI - Bilateral dentigerous cysts: review of the literature and report of an unusual case. AB - Dentigerous cysts are common cysts of the jaws. They are associated with the crowns of permanent teeth, most frequently associated with impacted mandibular third molars. Bilateral dentigerous cysts are rare and generally occur in association with a developmental syndrome or systemic disease, such as mucopolysaccharidosis and cleidocranial dysplasia. Bilateral dentigerous cysts in the absence of a syndrome are rare and to date only 17 cases have been described. The following is a report of a case of unusual bilateral non-syndromic dentigerous cysts associated with mandibular second molar and maxillary third molar and a review of the literature. Bilateral dentigerous cysts in these locations have not been previously reported. PMID- 17082342 TI - Buccal and palatal talon cusps with pulp extensions on a supernumerary primary tooth. AB - This paper reports an unusual occurrence of talon cusp on a supernumerary primary incisor, presenting on both labial and palatal sides. The tooth was scheduled for extraction due to its interference with the occlusion. Morphometric analysis of the taloned cusps was performed on digitized replicas of the tooth crown using open-source image analysis toolkit (ImageJ). Further non-destructive investigation of the taloned crown under cone-beam X-ray computed tomography revealed pulpal extensions in both talon cusps. PMID- 17082343 TI - Incidental radiological findings of susuk in the orofacial region. AB - Susuk, or charm needles, are inserted and worn subcutaneously in the face and other parts of the body, as they are believed to enhance beauty and youth, and for many other reasons such as treatment of headache, aches and pains in the joints, back or abdomen. The practice of inserting susuk is a traditional belief, genuinely cultural and superstitious, and common in the south-east Asian region. We present 13 cases of susuk, which was found incidentally on the radiographs as the patients came for various types of treatment at our centre. PMID- 17082344 TI - New nanophosphor scintillators for solid-state digital dental imagers. PMID- 17082345 TI - Muscle, liver, and pancreas: Three Musketeers fighting to control glycemia. PMID- 17082346 TI - Expression of the breast cancer resistance protein (Bcrp1/Abcg2) in tissues from pregnant mice: effects of pregnancy and correlations with nuclear receptors. AB - The breast cancer resistance protein (BCRP) plays an important role in drug disposition, including limiting drug penetration across the placental barrier. Our goal was to investigate the effects of pregnancy on Bcrp1 expression in pregnant mice. We examined Bcrp1 expression in placenta, kidney, liver, and small intestine at various gestational ages. Bcrp1 protein levels peaked at gestation day (gd) 15 in placenta, at gd 10 and 15 in kidney, and at gd 15 in liver; however, Bcrp1 protein levels in small intestine did not change significantly with gestational ages. Immunohistochemistry analysis revealed that the cellular localization of Bcrp1 in placenta, kidney, liver, and small intestine was not influenced by pregnancy. Bcrp1 mRNA levels were analyzed by quantitative real time RT-PCR. In general, the effects of pregnancy on Bcrp1 protein somewhat lagged behind the effects on Bcrp1 mRNA. To further investigate the possible roles of nuclear receptors in the regulation of the Bcrp1 gene during pregnancy, we examined mRNA levels of aryl hydrocarbon receptor (AhR), hypoxia-inducible factor 1alpha (HIF1alpha), estrogen receptor alpha (ERalpha), estrogen receptor beta (ERbeta), or progesterone receptor and compared them with those of Bcrp1. Bcrp1 mRNA was significantly correlated with mRNA of AhR, HIF1alpha, and ERbeta in placenta, with mRNA of HIF1alpha in kidney, and with mRNA of AhR and ERalpha in liver. These data suggest that Bcrp1 expression in mouse tissues can be altered by pregnancy in a gestational age-dependent manner. Such effects are likely mediated by certain nuclear receptors through a transcriptional mechanism. PMID- 17082347 TI - Significance of urea transport: the pioneering studies of Bodil Schmidt-Nielsen. PMID- 17082348 TI - Molecular modulation of calcium oxalate crystallization. AB - Calcium oxalate monohydrate (COM) is the primary constituent of the majority of renal stones. Osteopontin (OPN), an aspartic acid-rich urinary protein, and citrate, a much smaller molecule, are potent inhibitors of COM crystallization at levels present in normal urine. Current concepts of the role of site-specific interactions in crystallization derived from studies of biomineralization are reviewed to provide a context for understanding modulation of COM growth at a molecular level. Results from in situ atomic force microscopy (AFM) analyses of the effects of citrate and OPN on growth verified the critical role of site specific interactions between these growth modulators and individual steps on COM crystal surfaces. Molecular modeling investigations of interactions of citrate with steps and faces on COM crystal surfaces provided links between the stereochemistry of interaction and the binding energy levels that underlie mechanisms of growth modification and changes in overall crystal morphology. The combination of in situ AFM and molecular modeling provides new knowledge that will aid rationale design of therapeutic agents for inhibition of stone formation. PMID- 17082349 TI - Progressive postnatal increases in Fos immunoreactivity in the forebrain and brain stem of rats after viscerosensory stimulation with lithium chloride. AB - Interoceptive signals have a powerful impact on the motivation and emotional learning of animals during stressful experiences. However, current insights into the organization of interoceptive pathways stem mainly from observation and manipulation of adults, and little is known regarding the functional development of viscerosensory signaling pathways. To address this, we have examined central neural activation patterns in rat pups after treatment with lithium chloride (LiCl), a malaise-inducing agent. Rat pups were injected intraperitoneally with 0.15 M LiCl or 0.15 M NaCl (2% body wt) on postnatal day (P)0, 7, 14, 21, or 28, perfused 60 to 90 min postinjection, and their brains assayed for Fos protein immunolabeling. Compared with saline treatment, LiCl increased Fos only slightly in the area postrema, nucleus of the solitary tract, and lateral parabrachial nucleus on P0. LiCl did not increase Fos above control levels in the central nucleus of the amygdala, bed nucleus of the stria terminalis (BNST), or paraventricular nucleus of the hypothalamus on P0 but did on P7 and later. Maximal Fos responses to LiCl were observed on P14 in all areas except the BNST, in which LiCl-induced Fos activation continued to increase through P28. These results indicate that central LiCl-sensitive interoceptive circuits in rats are not fully functional at birth, and show age-dependent increases in neural Fos responses to viscerosensory stimulation with LiCl. PMID- 17082350 TI - Myofilament calcium sensitivity does not affect cross-bridge activation relaxation kinetics. AB - We employed single myofibril techniques to test whether the presence of slow skeletal troponin-I (ssTnI) is sufficient to induce increased myofilament calcium sensitivity (EC(50)) and whether modulation of EC(50) affects the dynamics of force development. Studies were performed using rabbit psoas myofibrils activated by rapid solution switch and in which Tn was partially replaced for either recombinant cardiac Tn(cTn) or Tn composed of recombinant cTn-T (cTnT) and cTn-C (cTnC), and recombinant ssTnI (ssTnI-chimera Tn). Tn exchange was performed in rigor solution (0.5 mg/ml Tn; 20 degrees C; 2 h) and confirmed by SDS-PAGE. cTnI exchange induced a decrease in EC(50); ssTnI-chimera Tn exchange induced a further decrease in EC(50) (in microM: endogenous Tn, 1.35 +/- 0.08; cTnI, 1.04 +/- 0.13; ssTnI-chimera Tn, 0.47 +/- 0.03). EC(50) was also decreased by application of 100 microM bepridil (control: 2.04 +/- 0.03 microM; bepridil 1.35 +/- 0.03 microM). Maximum tension was not different between any groups. Despite marked alterations in EC(50), none of the dynamic activation-relaxation parameters were affected under any condition. Our results show that 1) incorporation of ssTnI into the fast skeletal sarcomere is sufficient to induce increased myofilament Ca(2+) sensitivity, and 2) the dynamics of actin-myosin interaction do not correlate with EC(50). This result suggests that intrinsic cross-bridge cycling rate is not altered by the dynamics of thin-filament activation. PMID- 17082351 TI - Endogenous cholecystokinin reduces food intake and increases Fos-like immunoreactivity in the dorsal vagal complex but not in the myenteric plexus by CCK1 receptor in the adult rat. AB - We hypothesized that endogenous CCK reduces food intake by activating the dorsal vagal complex (DVC) and the myenteric neurons of the gut. To test this hypothesis, adult rats were given camostat mesilate; a nonnutrient releaser of endogenous CCK, by orogastric gavage, and Fos-like immunoreactivity (Fos-LI) was quantified in the DVC and the myenteric plexus. The results for endogenous CCK were compared with those for exogenous CCK-8. Exogenous CCK-8 reduced food intake and stimulated Fos-LI in the DVC and in myenteric neurons of the duodenum and jejunum. In comparison, endogenous CCK reduced food intake and increased DVC Fos LI but did not increase Fos-LI in the myenteric plexus. Similar to CCK-8, devazepide, a specific CCK(1) receptor antagonist, and not L365,260, a specific CCK(2) receptor antagonist, attenuated the reduction of food intake by camostat. In addition, Fos-LI in the DVC in response to both exogenous CCK-8 and camostat administration was significantly attenuated by vagotomy, as well as by blocking CCK(1) receptors. These results demonstrate for the first time that reduction of food intake in adult rats by endogenous CCK released by a nonnutrient mechanism requires CCK(1) receptors, the vagus nerve, and activation of the DVC, but not the myenteric plexus. PMID- 17082352 TI - Sex differences in postnatal growth and renal development in offspring of rabbit mothers with chronic secondary hypertension. AB - Previously, we demonstrated that adult blood pressure was increased in offspring of rabbit mothers with chronic secondary renal hypertension. Our study identified sex-specific differences in the programming of hypertension, with female, not male, offspring, having increased blood pressure at 30 wk of age. The aim of this study was to characterize the maternal hypertension during pregnancy to determine potential programming stimuli. Further, we examined the impact of chronic maternal hypertension on offspring birth weight, nephron number, and renal noradrenaline content (as an index of renal innervation density). Three groups of mothers and their offspring were studied: two-kidney, one-wrap (2K-1W, n = 9 mothers) hypertensive, two-kidney, two-wrap (2K-2W, n = 8) hypertensive, and a sham-operated group (n = 9). Mean arterial blood pressure was increased by approximately 20 mmHg throughout pregnancy in both hypertensive groups compared with sham mothers (P(G) < 0.001). Plasma renin activity (PRA; P(G) < 0.05) and aldosterone (P(G) < 0.05) levels were increased during gestation in the 2K-1W, but not the 2K-2W mothers. Birth weight was increased by approximately 20% in offspring of both groups of hypertensive mothers (P(T) < 0.001), though this was associated with a reduction in litter size. Renal noradrenaline content was increased ( approximately 40%, P < 0.05) at 5 wk of age in female 2K-1W offspring compared with sham offspring. Glomerular number was not reduced in female offspring of either group of hypertensive mothers; however, glomerular tuft volume was reduced in female 2K-2W offspring (P < 0.05), indicative of a reduction in glomerular filtration surface area. In conclusion, the two models of renal hypertension produced differential effects on the offspring. The impact of a stimulated maternal renin-angiotensin system in the 2K-1W model of hypertension may influence development of the renal sympathetic nerves and contribute to programming of adult hypertension. PMID- 17082353 TI - Chronic intermittent but not constant hypoxia decreases NAA/Cr ratios in neonatal mouse hippocampus and thalamus. AB - Chronic constant hypoxia (CCH) and chronic intermittent hypoxia (CIH) are known to have deleterious effects on the central nervous system. Because of the difference in the pattern of hypoxic exposure, it is possible that the pathological outcome would vary. The N-acetyl aspartate/creatine (NAA/Cr) ratio is a reliable marker of neuronal integrity, and this can be noninvasively measured by proton nuclear magnetic resonance spectroscopy. P2 CD1 mouse pups with their dams were exposed to either CCH, where the Fi(O(2)) was maintained at 11% continuously or to CIH, where the Fi(O(2)) was varied between 21 and 11% every 4 min. P30 mice exposed to intermittent hypoxia for 4 wk demonstrated a significant decrease in the NAA/Cr ratio in the hippocampus and thalamus, which was reversed by a subsequent exposure to 4 wk of normoxia. Meanwhile, mice exposed to 4 wk of constant hypoxia did not demonstrate any differences in their NAA/Cr ratios from controls in these brain regions. These results indicate that an intermittent pattern of hypoxic exposure may have a more adverse effect on neuronal function and integrity than a continuous one. The reversal of NAA/Cr levels to baseline during the return to normoxia indicates that therapeutic strategies targeted at alleviating the intermittent hypoxic stress in diseases, such as obstructive sleep apnea, have the potential for inducing significant neurocognitive recovery in these patients. PMID- 17082354 TI - Behavioral and neurochemical sources of variability of circadian period and phase: studies of circadian rhythms of npy-/- mice. AB - The cycle length or period of the free-running rhythm is a key characteristic of circadian rhythms. In this study we verify prior reports that locomotor activity patterns and running wheel access can alter the circadian period, and we report that these treatments also increase variability of the circadian period between animals. We demonstrate that the loss of a neurochemical, neuropeptide Y (NPY), abolishes these influences and reduces the interindividual variability in clock period. These behavioral and environmental influences, from daily distribution of peak locomotor activity and from access to a running wheel, both act to push the mean circadian period to a value < 24 h. Magnitude of light-induced resetting is altered as well. When photoperiod was abruptly changed from a 18:6-h light-dark cycle (LD18:6) to LD6:18, mice deficient in NPY were slower to respond to the change in photoperiod by redistribution of their activity within the prolonged dark and eventually adopted a delayed phase angle of entrainment compared with controls. These results support the hypothesis that nonphotic influences on circadian period serve a useful function when animals must respond to abruptly changing photoperiods and point to the NPYergic pathway from the intergeniculate leaflet innervating the suprachiasmatic nucleus as a circuit mediating these effects. PMID- 17082355 TI - The contribution of carotid rete variability to brain temperature variability in sheep in a thermoneutral environment. AB - The degree of variability in the temperature difference between the brain and carotid arterial blood is greater than expected from the presumed tight coupling between brain heat production and brain blood flow. In animals with a carotid rete, some of that variability arises in the rete. Using thermometric data loggers in five sheep, we have measured the temperature of arterial blood before it enters the carotid rete and after it has perfused the carotid rete, as well as hypothalamic temperature, every 2 min for between 6 and 12 days. The sheep were conscious, unrestrained, and maintained at an ambient temperature of 20-22 degrees C. On average, carotid arterial blood and brain temperatures were the same, with a decrease in blood temperature of 0.35 degrees C across the rete and then an increase in temperature of the same magnitude between blood leaving the rete and the brain. Rete cooling of arterial blood took place at temperatures below the threshold for selective brain cooling. All of the variability in the temperature difference between carotid artery and brain was attributable statistically to variability in the temperature difference across the rete. The temperature difference between arterial blood leaving the rete and the brain varied from -0.1 to 0.9 degrees C. Some of this variability was related to a thermal inertia of the brain, but the majority we attribute to instability in the relationship between brain blood flow and brain heat production. PMID- 17082356 TI - Novel axonal projection from the caudal end of the ventrolateral medulla to the intermediolateral cell column. AB - We used an optical imaging technique to investigate whether axons of neurons in the caudal end of the ventrolateral medulla (CeVLM), as well as axons of neurons in the rostral ventrolateral medulla (RVLM), project to neurons in the intermediolateral cell column (IML) of the spinal cord. Brain stem-spinal cord preparations from neonatal normotensive Wistar-Kyoto and spontaneously hypertensive rats were stained with a voltage-sensitive dye, and responses to electrical stimulation of the IML at the Th2 level were detected as changes in fluorescence intensity with an optical imaging apparatus (MiCAM-01). The results were as follows: 1) depolarizing responses to IML stimulation during low-Ca high Mg superfusion were detected on the ventral surface of the medulla at the level of the CeVLM, as well as at the level of the RVLM, 2) depolarizing responses were also detected on cross sections at the level of the CeVLM, and they had a latency of 24.0 +/- 5.5 (SD) ms, 3) antidromic action potentials in response to IML stimulation were demonstrated in the CeVLM neurons where optical images were detected, and 4) glutamate application to the CeVLM increased the frequency of excitatory postsynaptic potentials (EPSPs) and induced depolarization of the IML neurons. The optical imaging findings suggested a novel axonal and functional projection from neurons in the CeVLM to the IML. The increase in EPSPs of the IML neurons in response to glutamate application suggests that the CeVLM participates in the regulation of sympathetic nerve activity and blood pressure and may correspond to the caudal pressor area. PMID- 17082357 TI - Blood pressure regulation in neurally intact human vs. acutely injured paraplegic and tetraplegic patients during passive tilt. AB - We investigated autonomic control of cardiovascular function in able-bodied (AB), paraplegic (PARA), and tetraplegic (TETRA) subjects in response to head-up tilt following spinal cord injury. We evaluated spectral power of blood pressure (BP), baroreflex sensitivity (BRS), baroreflex effectiveness index (BEI), occurrence of systolic blood pressure (SBP) ramps, baroreflex sequences, and cross-correlation of SBP with heart rate (HR) in low (0.04-0.15 Hz)- and high (0.15-0.4 Hz) frequency regions. During tilt, AB and PARA effectively regulated BP and HR, but TETRA did not. The numbers of SBP ramps and percentages of heartbeats involved in SBP ramps and baroreflex sequences increased in AB, were unchanged in PARA, and declined in TETRA. BRS was lowest in PARA and declined with tilt in all groups. BEI was greatest in AB and declined with tilt in all groups. Low-frequency power of BP and the peak of the SBP/HR cross-correlation magnitude were greatest in AB, increased during tilt in AB, remained unchanged in PARA, and declined in TETRA. The peak cross-correlation magnitude in HF decreased with tilt in all groups. Our data indicate that spinal cord injury results in decreased stimulation of arterial baroreceptors and less engagement of feedback control as demonstrated by lower 1) spectral power of BP, 2) number (and percentages) of SBP ramps and barosequences, 3) cross-correlation magnitude of SBP/HR, 4) BEI, and 5) changes in delay between SBP/HR. Diminished vasomotion and impaired baroreflex regulation may be major contributors to decreased orthostatic tolerance following injury. PMID- 17082358 TI - Role of lipid rafts in membrane delivery of renal epithelial Na+-K+-ATPase, thick ascending limb. AB - Lipid rafts are cholesterol- and shingolipid-enriched membrane microdomains implicated in membrane signaling and trafficking. To assess renal epithelial raft functions through the characterization of their associated membrane proteins, we have isolated lipid rafts from rat kidney by sucrose gradient fractionation after detergent treatment. The low-density fraction was enriched in cholesterol, sphingolipid, and flotillin-1 known as lipid raft markers. Based on proteomic analysis of the low-density fraction, the protein with the highest significance score was the alpha-subunit of Na(+)-K(+)-ATPase (NKA), whose raft association was validated by simultaneous immunoblotting. The beta-subunit of NKA was copurified from the low-density fraction. To test the role of lipid rafts in sorting and membrane delivery of renal-transporting epithelia, we have chosen to study thick ascending limb (TAL) epithelium for its high NKA activity and the property to be stimulated by antidiuretic hormone (ADH). Cultured rabbit TAL cells were studied. Cholesterol depletion and detergent extraction at warmth caused a shift of NKA to the higher-density fractions. Comparative preparations from blood monocytes revealed the absence of NKA from rafts in these nonpolarized cells. Short-term exposure of rabbit TAL cells to ADH (1 h) caused translocation and enhanced raft association of NKA via cAMP activation. Preceding cholesterol depletion prevented this effect. TAL-specific, glycosylphosphatidylinositol anchored Tamm Horsfall protein was copurified with NKA in the same raft fraction, suggesting functional interference between these products. These results may have functional implications regarding the turnover, trafficking, and regulated surface expression of NKA as the major basolateral ion transporter of TAL. PMID- 17082360 TI - Contributions of passive mechanical loads and active neuromuscular compensation to upper airway collapsibility during sleep. PMID- 17082361 TI - Comments on point: counterpoint: "cardiovascular variability is/is not an index of autonomic control of circulation". PMID- 17082362 TI - Endothelin receptor blockade does not improve hypoxemia following acute pulmonary thromboembolism. AB - We studied the roles of endothelins in determining ventilation (Va) and perfusion (Q) mismatch in a porcine model of acute pulmonary thromboembolism (APTE), using a nonspecific endothelin antagonist, tezosentan. Nine anesthetized piglets (approximately 23 kg) received autologous clots (approximately 20 g) via a central venous catheter at time = 0 min. The distribution of Va and Q at five different time points (-30, -5, 30, 60, 120 min) was mapped by fluorescent microspheres of 10 different colors. Five piglets (group 1) received tezosentan (courtesy of Actelion) starting at time = 40 min for 2 h, and four piglets (group 2) received only saline and served as control. Our results showed that, in all of the animals at 30 min following APTE but before tezosentan, the mean Va/Q was increased, as was Va/Q heterogeneity (log SD Va/Q), which represented a widening of its main peak. Afterwards, tezosentan attenuated the pulmonary hypertension in group 1 but also produced moderate systemic hypotension. However, it did not improve arterial PO2 or Va/Q mismatch. We concluded that endothelin antagonism had minimal impact on gas exchange following APTE and confirmed our earlier observation that the main mechanism for hypoxemia in APTE was due to the mechanical redistribution of pulmonary regional blood flow away from the embolized vessels, resulting in the creation of many divergent low and high Va/Q regions. PMID- 17082363 TI - Energy sensing and regulation of gene expression in skeletal muscle. AB - Major modifications in energy homeostasis occur in skeletal muscle during exercise. Emerging evidence suggests that changes in energy homeostasis take part in the regulation of gene expression and contribute to muscle plasticity. A number of energy-sensing molecules have been shown to sense variations in energy homeostasis and trigger regulation of gene expression. The AMP-activated protein kinase, hypoxia-inducible factor 1, peroxisome proliferator-activated receptors, and Sirt1 proteins all contribute to altering skeletal muscle gene expression by sensing changes in the concentrations of AMP, molecular oxygen, intracellular free fatty acids, and NAD+, respectively. These molecules may therefore sense information relating to the intensity, duration, and frequency of muscle exercise. Mitochondria also contribute to the overall response, both by modulating the response of energy-sensing molecules and by generating their own signals. This review seeks to examine our current understanding of the roles that energy-sensing molecules and mitochondria can play in the regulation of gene expression in skeletal muscle. PMID- 17082364 TI - The production of reactive oxygen and nitrogen species by skeletal muscle. AB - Skeletal muscle has been recognized as a potential source for generation of reactive oxygen and nitrogen species for more than 20 years. Initial investigations concentrated on the potential role of mitochondria as a major source for generation of superoxide as a "by-product" of normal oxidative metabolism, but recent studies have identified multiple subcellular sites, where superoxide or nitric oxide are generated in regulated and controlled systems in response to cellular stimuli. Full evaluation of the factors regulating these processes and the functions of the reactive oxygen species generated are important in understanding the redox biology of skeletal muscle. PMID- 17082365 TI - Hyperlipidemia and lipid peroxidation are dependent on the severity of chronic intermittent hypoxia. AB - Obstructive sleep apnea (OSA) is characterized by chronic intermittent hypoxia (CIH) and associated with dysregulation of lipid metabolisms and atherosclerosis. Causal relationships between OSA and metabolic abnormalities have not been established because of confounding effects of underlying obesity. The goal of the study was to determine if CIH causes lipid peroxidation and dyslipidemia in the absence of obesity and whether the degrees of dyslipidemia and lipid peroxidation depend on the severity of hypoxia. Lean C57BL/6J mice were exposed to CIH for 4 wk with a fractional inspired O2 (FI(O2)) nadir of either 10% (moderate CIH) or 5% (severe CIH). Mice exposed to severe CIH exhibited significant increases in fasting serum levels of total cholesterol (129 +/- 2.9 vs. 113 +/- 2.8 mg/dl in control mice, P < 0.05) and low-density lipoprotein cholesterol (85.7 +/- 8.9 vs. 56.4 +/- 9.7 mg/dl, P < 0.05) in conjunction with a 1.5- to 2-fold increase in lipoprotein secretion, and upregulation of hepatic stearoyl coenzyme A desaturase 1 (SCD-1). Severe CIH also markedly increased lipid peroxidation in the liver (malondialdehyde levels of 94.4 +/- 5.4 vs. 57.4 +/- 5.2 nmol/mg in control mice, P < 0.001). In contrast, moderate CIH did not induce hyperlipidemia or change in hepatic SCD-1 levels but did cause lipid peroxidation in the liver at a reduced level relative to severe CIH. In conclusion, CIH leads to hypercholesterolemia and lipid peroxidation in the absence of obesity, and the degree of metabolic dysregulation is dependent on the severity of the hypoxic stimulus. PMID- 17082366 TI - Modulation of glucose transport in skeletal muscle by reactive oxygen species. AB - Glucose transport is an essential physiological process that is characteristic of all eukaryotic cells, including skeletal muscle. In skeletal muscle, glucose transport is mediated by the GLUT-4 protein under conditions of increased carbohydrate utilization. The three major physiological stimuli of glucose transport in muscle are insulin, exercise/contraction, and hypoxia. Here, the role of reactive oxygen species (ROS) in modulating glucose transport in skeletal muscle is reviewed. Convincing evidence for ROS involvement in insulin- and hypoxia-mediated transport in muscle is lacking. Recent experiments, based on pharmacological and genetic approaches, support a role for ROS in contraction mediated glucose transport. During contraction, endogenously produced ROS appear to mediate their effects on glucose transport via AMP-activated protein kinase. PMID- 17082367 TI - Acute sympathetic vasoconstriction at rest and during dynamic exercise in cyclists and sedentary humans. AB - The impact of exercise training on sympathetic activation is not well understood, especially across untrained and trained limbs in athletes. Therefore, in eight sedentary subjects (maximal oxygen consumption = 40 +/- 2 ml x kg(-1) x min(-1)) and eight competitive cyclists (maximal oxygen consumption = 64 +/- 2 ml x kg(-1) x min(-1)), we evaluated heart rate, blood pressure, blood flow, vascular conductance, and vascular resistance in the leg and arm during acute sympathetic stimulation [cold pressor test (CPT)]. The CPT was also performed during dynamic leg (knee extensor) or arm (handgrip) exercise at 50% of maximal work rate (WRmax) with measurements in the exercising limb. At rest, the CPT decreased vascular conductance similarly in the leg and arm of sedentary subjects (-33 +/- 8% leg, -38 +/- 6% arm) and cyclists (-34 +/- 4% leg, -31 +/- 9% arm), and during exercise CPT-induced vasoconstriction was blunted (i.e., sympatholysis) in both the leg and arm of both groups. However, the magnitude of sympatholysis was significantly different between the arm and leg of the sedentary group (-47 +/- 11% arm, -25 +/- 8% leg), and it was less in the arm of cyclists (-28 +/- 11%) than sedentary controls. Taken together, these data provide evidence that sympathetically mediated vasoconstriction is expressed equally and globally at rest in both sedentary and trained individuals, with a differential pattern of vasoconstriction during acute exercise according to limb and exercise training status. PMID- 17082368 TI - Microgravity-induced changes in aortic stiffness and their role in orthostatic intolerance. AB - Microgravity (microG)-induced orthostatic intolerance (OI) in astronauts is characterized by a marked decrease in cardiac output (CO) in response to an orthostatic stress. Since CO is highly dependent on venous return, alterations in the resistance to venous return (RVR) may be important in contributing to OI. The RVR is directly dependent on arterial compliance (C(a)), where aortic compliance (C(ao)) contributes up to 60% of C(a). We tested the hypothesis that microG induced changes in C(a) may represent a protective mechanism against OI. A retrospective analysis on hemodynamic data collected from astronauts after 5- to 18-day spaceflight missions revealed that orthostatically tolerant (OT) astronauts showed a significant decrease in C(a) after spaceflight, while OI astronauts showed a slight increase in C(a). A ground-based animal model simulating microG, hindlimb-unweighted rats, was used to explore this phenomenon. Two independent assessments of C(ao), in vivo pulse wave velocity (PWV) of the thoracic aorta and in vitro pressure-diameter squared relationship (PDSR) measurements of the excised thoracic aorta, were determined. PWV showed a significant increase in aortic stiffness compared with control, despite unchanged blood pressures. This increase in aortic stiffness was confirmed by the PDSR analysis. Thus both actual microG in humans and simulated microG in rats induces changes in C(ao). The difference in C(a) in OT and OI astronaut suggests that the microG-induced decrease in C(a) is a protective adaptation to spaceflight that reduces the RVR and allows for the maintenance of adequate CO in response to an orthostatic stress. PMID- 17082369 TI - Uncoupling of upper airway motor activity from phrenic bursting by positive end expired pressure in the rat. AB - Phasic bursting in the hypoglossal nerve can be uncoupled from phrenic bursting by application of positive end-expired pressure (PEEP). We wished to determine whether similar uncoupling can also be induced in other respiratory-modulated upper airway (UAW) motor outputs. Discharge of the facial, hypoglossal, superior laryngeal, recurrent laryngeal, and phrenic nerves was recorded in anesthetized, ventilated rats during stepwise changes in PEEP with a normocapnic, hyperoxic background. Application of 3- to 6-cmH(2)O PEEP caused the onset inspiratory (I) UAW nerve bursting to precede the phrenic burst but did not uncouple bursting. In contrast, application of 9- to 12-cmH(2)O PEEP uncoupled UAW neurograms such that rhythmic bursting occurred during periods of phrenic quiescence. Single-fiber recording experiments were conducted to determine whether a specific population of UAW motoneurons is recruited during uncoupled bursting. The data indicate that expiratory-inspiratory (EI) motoneurons remained active, while I motoneurons did not fire during uncoupled UAW bursting. Finally, we examined the relationship between motoneuron discharge rate and PEEP during coupled UAW and phrenic bursting. EI discharge rate was linearly related to PEEP during preinspiration, but showed no relationship to PEEP during inspiration. Our results demonstrate that multiple UAW motor outputs can be uncoupled from phrenic bursting, and this response is associated with bursting of EI nerve fibers. The relationship between PEEP and EI motoneuron discharge rate differs during preinspiratory and I periods; this may indicate that bursting during these phases of the respiratory cycle is controlled by distinct neuronal outputs. PMID- 17082370 TI - Apparatus for measuring rat body volume: a methodological proposition. AB - We propose a communicating-vessels system to measure body volume in live rats through water level detection by hydrostatic weighing. The reproducibility, accuracy, linearity, and reliability of this apparatus were evaluated in two tests using previously weighed water or six aluminum cylinders of known volume after proper system calibration. The applicability of this apparatus to measurement of live animals (Wistar rats) was tested in a transversal experiment with five rats, anesthetized and nonanesthetized. We took 18 measurements of the volume under each condition (anesthetized and nonanesthetized), totaling 90 measurements. The addition of water volumes (50-700 ml) produced a regression equation with a slope of 1.0006 +/- 0.0017, intercept of 0.75 +/- 0.81 (R(2) = 0.99999, standard error of estimate = 0.58 ml), and bias of approximately 1 ml. The differences between cylinders of known volumes and volumes calculated by the system were <0.4 ml. Mean volume errors were 0.01-0.07%. Among the live models, the difference between the volumes obtained for anesthetized and nonanesthetized rats was 0.31 +/- 2.34 (SD) ml (n = 90). These data showed that animal movement does not interfere with the volume measured by the proposed apparatus, and neither anesthesia nor fur shaving is needed for this procedure. Nevertheless, some effort should be taken to eliminate air bubbles trapped in the apparatus or the fur. The proposed apparatus for measuring rat body volume is inexpensive and may be useful for a range of scientific purposes. PMID- 17082371 TI - Sex differences in left ventricular function and beta-receptor responsiveness following prolonged strenuous exercise. AB - Sex differences in neuroendocrine and metabolic responses to prolonged strenuous exercise (PSE) have been well documented. The aim of this investigation was to examine sex differences in left ventricular function and cardiac beta-receptor responsiveness following a single bout of PSE. Nine male and eight female triathletes were examined during three separate sessions: before, immediately after, and 24 h following a half-ironman triathlon using dobutamine stress echocardiography. Steady-state graded infusions of dobutamine were used to assess beta-receptor responsiveness. Slopes calculated from linear regressions between dobutamine doses and changes in heart rate and contractility for each participant were used as an index of beta-receptor responsiveness. Despite no change in preload, fractional area change decreased from baseline after the race in both men and women, with a greater decrease in men [men: 54.1% (SD 2.1) to 50.7% (SD 3.4) vs. women: 55.4% (SD 2.7) to 53.3% (SD 2.5); P < 0.05]. The amount of dobutamine necessary to increase heart rate by 25 beats/min [men: 29.6 microg x kg(-1) x min(-1) (SD 6.6) to 42.7 microg x kg(-1) x min(-1) (SD 12.9) vs. women: 23.5 microg x kg(-1) x min(-1) (SD 4.0) to 30.0 microg x kg(-1) x min(-1) (SD 7.8); P < 0.05] and contractility by 10 mmHg/cm2 [men: 20.9 microg x kg(-1) x min(-1) (SD 5.1) to 37.0 microg x kg(-1) x min(-1) (SD 11.5) vs. women: 22.6 microg x kg(-1) x min(-1) (SD 6.4) to 30.7 microg x kg(-1) x min(-1) (SD 7.2); P < 0.05] was greater in both men and women postrace. However, the amount of dobutamine required to induce these changes was greater in men, reflecting larger beta-receptor alterations in male triathletes following PSE relative to women. These data suggest that following an acute bout of PSE, male triathletes demonstrate an attenuated chronotropic and inotropic response to beta-adrenergic stimulation compared with female triathletes. PMID- 17082372 TI - Hypothermia reduces ischemia- and stimulation-induced myocardial interstitial norepinephrine and acetylcholine releases. AB - Although hypothermia is one of the most powerful modulators that can reduce ischemic injury, the effects of hypothermia on the function of the cardiac autonomic nerves in vivo are not well understood. We examined the effects of hypothermia on the myocardial interstitial norepinephrine (NE) and ACh releases in response to acute myocardial ischemia and to efferent sympathetic or vagal nerve stimulation in anesthetized cats. We induced acute myocardial ischemia by coronary artery occlusion. Compared with normothermia (n = 8), hypothermia at 33 degrees C (n = 6) suppressed the ischemia-induced NE release [63 nM (SD 39) vs. 18 nM (SD 25), P < 0.01] and ACh release [11.6 nM (SD 7.6) vs. 2.4 nM (SD 1.3), P < 0.01] in the ischemic region. Under hypothermia, the coronary occlusion increased the ACh level from 0.67 nM (SD 0.44) to 6.0 nM (SD 6.0) (P < 0.05) and decreased the NE level from 0.63 nM (SD 0.19) to 0.40 nM (SD 0.25) (P < 0.05) in the nonischemic region. Hypothermia attenuated the nerve stimulation-induced NE release from 1.05 nM (SD 0.85) to 0.73 nM (SD 0.73) (P < 0.05, n = 6) and ACh release from 10.2 nM (SD 5.1) to 7.1 nM (SD 3.4) (P < 0.05, n = 5). In conclusion, hypothermia attenuated the ischemia-induced NE and ACh releases in the ischemic region. Moreover, hypothermia also attenuated the nerve stimulation induced NE and ACh releases. The Bezold-Jarisch reflex evoked by the left anterior descending coronary artery occlusion, however, did not appear to be affected under hypothermia. PMID- 17082373 TI - High-intensity exercise acutely decreases the membrane content of MCT1 and MCT4 and buffer capacity in human skeletal muscle. AB - The regulation of intracellular pH during intense muscle contractions occurs via a number of different transport systems [e.g., monocarboxylate transporters (MCTs)] and via intracellular buffering (beta m(in vitro)). The aim of this study was to investigate the effects of an acute bout of high-intensity exercise on both MCT relative abundance and beta m(in vitro) in humans. Six active women volunteered for this study. Biopsies of the vastus lateralis were obtained at rest and immediately after 45 s of exercise at 200% of maximum O2 uptake. Beta m(in vitro) was determined by titration, and MCT relative abundance was determined in membrane preparations by Western blots. High-intensity exercise was associated with a significant decrease in both MCT1 (-24%) and MCT4 (-26%) and a decrease in beta m(in vitro) (-11%; 135 +/- 3 to 120 +/- 2 micromol H+ x g dry muscle(-1) x pH(-1); P < 0.05). These changes were consistently observed in all subjects, and there was a significant correlation between changes in MCT1 and MCT4 relative abundance (R2 = 0.92; P < 0.05). In conclusion, a single bout of high-intensity exercise decreased both MCT relative abundance in membrane preparations and beta m(in vitro). Until the time course of these changes has been established, researchers should consider the possibility that observed training-induced changes in MCT and beta m(in vitro) may be influenced by the acute effects of the last exercise bout, if the biopsy is taken soon after the completion of the training program. The implications that these findings have for lactate (and H+) transport following acute, exhaustive exercise warrant further investigation. PMID- 17082374 TI - Exercise training improves cardiac performance in diabetes: in vivo demonstration with quantitative cine-MRI analyses. AB - Diabetic cardiomyopathy is a distinct myocardial complication of the catabolic state of untreated insulin-dependent diabetes mellitus in the streptozotocin induced diabetic rat. Exercise training has long been utilized as an effective adjunct to pharmacotherapy in the management of the diabetic heart. However, the in vivo functional benefit(s) of the training programs on cardiac cycle events in diabetes are poorly understood. In this study, we used three groups of Sprague Dawley rats (sedentary control, sedentary diabetic, and exercised diabetic) to assess the effects of endurance training on the left ventricular (LV) cardiac cycle events in diabetes. At the end of 9 wk of exercise training, noninvasive cardiac functional evaluation was performed by using high-resolution magnetic resonance imaging (9.4 T). An ECG-gated cine imaging protocol was used to capture the LV cardiac cycle events through 10 equally incremented phases. The cardiac cycle phase volumetric profiles showed favorable functional changes in exercised diabetic group, including a prevention of decreased end-diastolic volume and attenuation of increased end-systolic volume that accompanies sedentary diabetes. The defects in LV systolic flow velocity, acceleration, and jerk associated with sedentary diabetes were restored toward control levels in the trained diabetic animals. This magnetic resonance imaging study confirms the prevailing evidence from earlier in vitro and in vivo invasive procedures that exercise training benefits cardiac function in this model of diabetic cardiomyopathy despite the extreme catabolic state of the animals. PMID- 17082375 TI - Electromyographic activity during the reflex pharyngeal swallow in the pig: Doty and Bosma (1956) revisited. AB - The currently accepted description of the pattern of electromyographic (EMG) activity in the pharyngeal swallow is that reported by Doty and Bosma in 1956; however, those authors describe high levels of intramuscle and of interindividual EMG variation. We reinvestigated this pattern, testing two hypotheses concerning EMG variation: 1) that it could be reduced with modern methodology and 2) that it could be explained by selective detection of different types of motor units. In eight decerebrate infant pigs, we elicited radiographically verified pharyngeal swallows and recorded EMG activity from a total of 16 muscles. Synchronization signals from the video-radiographic system allowed the EMG activity associated with each swallow to be aligned directly with epiglottal movement. The movements were highly stereotyped, but the recorded EMG signals were variable at both the intramuscle and interanimal level. During swallowing, some muscles subserved multiple functions and contained different task units; there were also intramuscle differences in EMG latencies. In this situation, statistical methods were essential to characterize the overall patterns of EMG activity. The statistically derived multimuscle pattern approximated to the classical description by Doty and Bosma (Doty RW, Bosma JF. J Neurophysiol 19: 44-60, 1956) with a leading complex of muscle activities. However, the mylohyoid was not active earlier than other muscles, and the geniohyoid muscle was not part of the leading complex. Some muscles, classically considered inactive, were active during the pharyngeal swallow. PMID- 17082376 TI - Effects of forskolin on inotropic performance and phospholamban phosphorylation in exercise-trained hypertensive myocardium. AB - Beta-adrenergic receptor (beta-AR) responsiveness is downregulated in left ventricular (LV) hypertrophy induced by chronic hypertension. While exercise training in hypertension enhances beta-AR responsiveness, the role of adenylyl cyclase remains unclear. The purpose of the present study was to test whether treadmill running in the spontaneously hypertensive rat (SHR) model improves LV responsiveness to forskolin (FOR) or the combination of FOR + isoproterenol (FOR+ISO). Female SHR (16-wk) were randomly placed into sedentary (SHR-SED; n = 7) or treadmill-trained (SHR-TRD; n = 8) groups. Wistar-Kyoto (WKY; n = 7) animals acted as normotensive controls. Langendorff, isovolumic LV performance was established at baseline and during incremental FOR infusion (1 and 5 micromol/l) and FOR+ISO (5 micromol/l + 1x10(-8) mol/l). Heart rate, systolic blood pressure, and heart-to-body weight ratio were lower in WKY relative to both SHR groups (P < 0.05). LV performance and heart rate significantly increased in all groups to a similar extent with incremental FOR infusion. However, in the presence of 5 micromol/l FOR, ISO increased LV developed pressure, positive change in LV pressure, and negative change in LV pressure to a greater extent in SHR-TRD relative to SHR-SED (P < 0.05). Phospholamban phosphorylation at the Thr17 was greater in SHR-TRD relative to SHR-SED and WKY (P < 0.05). Absolute LV developed pressure was moderately correlated with phospholamban phosphorylation at both the Ser16 (r = 0.64; P < 0.05) and Thr17 (r = 0.52; P < 0.05). Our data suggest that the adenylyl cyclase step in the beta-AR cascade is not downregulated in the early course of hypertension and that the enhanced beta-AR responsiveness with training is likely mediated at levels other than adenylyl cyclase. Our data also suggest that beta-AR inotropic responsiveness in the presence of direct adenylyl cyclase agonism is improved in trained compared with sedentary SHR hearts. PMID- 17082377 TI - The effect of exercise on nasal uptake of ozone in healthy human adults. AB - The nose may help protect the lower respiratory tract from the effects of ambient ozone by scrubbing ozone from inspired air. Reductions in both nasal resistance and nitric oxide production with exercise may influence the efficiency of ozone uptake in the nose. Nasal ozone uptake was measured in 10 healthy volunteers before and after 15 min of moderate bicycle exercise. Ozone (0.2 parts/million) was pulled through both nostrils and out of the mouth at a constant flow while the subjects closed their epiglottises. Nasal uptake of ozone was determined by comparing the ozone concentration entering the nostrils to that exiting the mouth. Average preexercise uptake of ozone was 56 +/- 7.8 and 37 +/- 4.9% at 10 and 20 l/min, respectively. These averages did not significantly differ from those immediately postexercise (55 and 37%). Nasal ozone uptake increased significantly (P < 0.001) with decreasing flow rate, but intersubject variability in uptake could not be predicted by nasal volume or cross-sectional areas (as measured by acoustic rhinometry) or endogenous nitric oxide production. However, the percent change in ozone uptake after exercise, within an individual, was correlated with both 1) percent change in nasal volume (r = 0.70 at 10 l/min) and 2) percent change in the rate of volumetric expansion between the nasal valve and turbinates (r = 0.82 at 10 l/min). These results may be useful for assessing human risk associated with ozone exposure during exercise. PMID- 17082378 TI - Endothelial dysfunction and decreased vascular responsiveness in the anterior cruciate ligament-deficient model of osteoarthritis. AB - Chronic inflammation associated with osteoarthritis (OA) may alter normal vascular responses and contribute to joint degradation. Vascular responses to vasoactive mediators were evaluated in the medial collateral ligament (MCL) of the anterior cruciate ligament (ACL)-deficient knee. Chronic joint instability and progressive OA were induced in rabbit knees by surgical transection of the ACL. Under halothane anesthesia, laser speckle perfusion imaging (LSPI) was used to measure MCL blood flow in unoperated control (n = 12) and 6-wk ACL-transected knees (n = 12). ACh, bradykinin, histamine, substance P (SP), and prostaglandin E(2) (PGE(2)) were applied to the MCL vasculature in topical boluses of 100 microl (dose range 10(-14) to 10(-8) mol). In normal joints, ACh, bradykinin, histamine, and PGE(2) evoked a dilatory response. Substance P caused a biphasic response that was dilatory from 10(-14) to 10(-11) mol and constricting at higher doses. In ACL-deficient knees, ACh, bradykinin, histamine, and SP decreased perfusion, whereas PGE(2) had a biphasic response that decreased perfusion at 10( 14) to 10(-11) mol and was dilatory at higher concentrations. Sodium nitroprusside increased perfusion in resting and phenylephrine-precontracted vessels with no significant differences between ACL-transected and control knees. Femoral artery occlusion and release increased perfusion by 74.3 +/- 11.1% in control knees but only by 25.8 +/- 4.4% in ACL-deficient knees. The altered responsiveness of the MCL vasculature to these inflammatory mediators may indicate endothelial dysfunction in the MCL, which may contribute to the progression and severity of OA and to the adaptation of the joint in an altered mechanical environment. PMID- 17082379 TI - Time course of ozone-induced changes in breathing pattern in healthy exercising humans. AB - We examined the time course of O3-induced changes in breathing pattern in 97 healthy human subjects (70 men and 27 women). One- to five-minute averages of breathing frequency (f(B)) and minute ventilation (Ve) were used to generate plots of cumulative breaths and cumulative exposure volume vs. time and cumulative exposure volume vs. cumulative breaths. Analysis revealed a three phase response; delay, no response detected; onset, f(B) began to increase; response, f(B) stabilized. Regression analysis was used to identify four parameters: time to onset, number of breaths at onset, cumulative inhaled dose of ozone at onset of O3-induced tachypnea, and the percent change in f(B). The effect of altering O3 concentration, Ve, atropine treatment, and indomethacin treatment were examined. We found that the lower the O3 concentration, the greater the number of breaths at onset of tachypnea at a fixed ventilation, whereas number of breaths at onset of tachypnea remains unchanged when Ve is altered and O3 concentration is fixed. The cumulative inhaled dose of O3 at onset of tachypnea remained constant and showed no relationship with the magnitude of percent change in f(B). Atropine did not affect any of the derived parameters, whereas indomethacin did not affect time to onset, number of breaths at onset, or cumulative inhaled dose of O3 at onset of tachypnea but did attenuate percent change in f(B). The results are discussed in the context of dose response and intrinsic mechanisms of action. PMID- 17082380 TI - List of new names and new combinations previously effectively, but not validly, published. PMID- 17082381 TI - Notification that new names and new combinations have appeared in volume 56, part 8, of the IJSEM. PMID- 17082382 TI - Marinobacter vinifirmus sp. nov., a moderately halophilic bacterium isolated from a wine-barrel-decalcification wastewater. AB - A halophilic, Gram-negative, motile, non-sporulating bacterium designated strain FB1T was isolated from a wine-barrel-decalcification wastewater. The organism comprises straight rods and has a strictly respiratory metabolism with O2. Strain FB1T grows optimally at 20-30 degrees C and 5-6% NaCl. The predominant fatty acids were found to be C18:1omega9c (30.4%), C16:0 (25.7%), C12:0 3-OH (10.3%), C16:1omega9c (9.7%) and C16:1omega7c (8.4%). A phylogenetic analysis based on 16S rRNA gene sequences revealed that the strain forms a coherent cluster within the genus Marinobacter. The highest level of 16S rRNA gene sequence similarity (97.9%) exhibited by strain FB1T was with the type strain of Marinobacter excellens. However, the level of DNA-DNA relatedness between the novel strain and M. excellens CIP 107,686T was only 31.2%. The DNA G+C content of strain FB1T was 58.7 mol%. On the basis of phenotypic and genotypic characteristics, and also phylogenetic evidence, strain FB1T is considered to represent a novel species of the genus Marinobacter, for which the name Marinobacter vinifirmus sp. nov. is proposed. The type strain is FB1T (=DSM 17747T=CCUG 52119T). PMID- 17082383 TI - Methyloversatilis universalis gen. nov., sp. nov., a novel taxon within the Betaproteobacteria represented by three methylotrophic isolates. AB - The taxonomic positions and phylogenetic relationships of two new methylotrophic isolates from Lake Washington (USA) sediment, FAM5T and 500, and the previously described methylotrophic strain EHg5 isolated from contaminated soil in Estarreja (Portugal) were investigated. All three strains were facultative methylotrophs capable of growth on a variety of C1 and multicarbon compounds. Optimal growth occurred at pH 7.5-8 and 30-37 degrees C. The major fatty acids were C16:1omega7c and C16:0. The major quinone was ubiquinone Q8. Neither methanol dehydrogenase nor methanol oxidase activities were detectable in cells grown on methanol, suggesting an alternative, as-yet unknown, mechanism for methanol oxidation. The isolates assimilated C1 units at the level of formaldehyde, via the serine cycle. The DNA G+C content of the strains ranged between 64 and 65 mol%. 16S rRNA gene sequence similarity between the three new isolates was 99.85-100%, but was below 94% with other members of the Betaproteobacteria, indicating that the isolates represent a novel taxon. Based on physiological, phenotypic and genomic characteristics of the three isolates, a new genus, Methyloversatilis gen. nov., is proposed within the family Rhodocyclaceae. The type strain of Methyloversatilis universalis gen. nov., sp. nov. is FAM5T (=CCUG 52030T=JCM 13912T). PMID- 17082384 TI - Reclassification of Lactobacillus amylophilus LMG 11400 and NRRL B-4435 as Lactobacillus amylotrophicus sp. nov. AB - The taxonomic position of six Lactobacillus amylophilus strains isolated from swine waste-corn fermentations was reinvestigated. All strains were included in a multilocus sequence analysis (MLSA) study for species identification of Lactobacillus using the genes encoding the phenylalanyl-tRNA synthase alpha subunit (pheS) and RNA polymerase alpha subunit (rpoA). Partial pheS and rpoA gene sequences showed that strains LMG 11400 and NRRL B-4435 represent a separate lineage that is distantly related to the type strain of L. amylophilus, LMG 6900T, and to three other strains of the species. The MLSA data showed that the two strains LMG 11400 and NRRL B-4435 constituted a distinct cluster, sharing 100% pheS and rpoA gene sequence similarity. The other reference strains clustered together with the type strain of L. amylophilus, LMG 6900T, and were clearly differentiated from strains LMG 11400 and NRRL B-4435 (80 and 89% pheS and rpoA gene sequence similarity, respectively). The 16S rRNA gene sequences of the latter two strains are 100% identical, with the nearest phylogenetic neighbour L. amylophilus LMG 6900T showing only 97.2% 16S rRNA gene sequence similarity. Further polyphasic taxonomic study based on whole-cell protein fingerprinting, DNA-DNA hybridization and biochemical features demonstrated that the two strains represent a single, novel Lactobacillus species, for which the name Lactobacillus amylotrophicus sp. nov. is proposed. The type strain is LMG 11400T (=NRRL B-4436T=DSM 20534T). PMID- 17082385 TI - Burkholderia ginsengisoli sp. nov., a beta-glucosidase-producing bacterium isolated from soil of a ginseng field. AB - A bacterial strain (designated KMY03T) that possesses beta-glucosidase activity was isolated from soil from a ginseng field in South Korea and was characterized in order to determine its taxonomic position. The bacterium was found to comprise Gram-negative, rod-shaped, motile cells with unipolar polytrichous flagella. On the basis of 16S rRNA gene sequence similarity, strain KMY03T was shown to belong to the family Burkholderiaceae of the Betaproteobacteria, being most closely related to Burkholderia caledonica LMG 19076T (97.8%), Burkholderia terricola LMG 20594T (97.5%), Burkholderia xenovorans LMG 21463T (97.4%) and Burkholderia phytofirmans LMG 22146T (97.3%). Chemotaxonomic data (major ubiquinone, Q-8; major fatty acids, C17:0 cyclo, C16:0, C19:0 cyclo omega8c and summed feature 2) supported the affiliation of the novel strain with the genus Burkholderia. The results of DNA-DNA hybridizations and physiological and biochemical tests allowed the strain to be differentiated genotypically and phenotypically from Burkholderia species with validly published names. On the basis of these data, strain KMY03T represents a novel species of the genus Burkholderia, for which the name Burkholderia ginsengisoli sp. nov. is proposed. The type strain is KMY03T (=KCTC 12389T=NBRC 100965T). PMID- 17082386 TI - 'Candidatus Midichloria mitochondrii', an endosymbiont of the tick Ixodes ricinus with a unique intramitochondrial lifestyle. AB - An intracellular bacterium with the unique ability to enter mitochondria exists in the European vector of Lyme disease, the hard tick Ixodes ricinus. Previous phylogenetic analyses based on 16S rRNA gene sequences suggested that the bacterium formed a divergent lineage within the Rickettsiales (Alphaproteobacteria). Here, we present additional phylogenetic evidence, based on the gyrB gene sequence, that confirms the phylogenetic position of the bacterium. Based on these data, as well as electron microscopy (EM), in situ hybridization and other observations, we propose the name 'Candidatus Midichloria mitochondrii' for this bacterium. The symbiont appears to be ubiquitous in females of I. ricinus across the tick's distribution, while lower prevalence is observed in males (44%). Based on EM and in situ hybridization studies, the presence of 'Candidatus M. mitochondrii' in females appears to be restricted to ovarian cells. The bacterium was found to be localized both in the cytoplasm and in the intermembrane space of the mitochondria of ovarian cells. 'Candidatus M. mitochondrii' is the first bacterium to be identified that resides within animal mitochondria. PMID- 17082387 TI - Bacillus infantis sp. nov. and Bacillus idriensis sp. nov., isolated from a patient with neonatal sepsis. AB - Two Gram-positive bacilli, designated as strains SMC 4352-1T and SMC 4352-2T, were isolated sequentially from the blood of a newborn child with sepsis. They could not be identified by using conventional clinical microbiological methods. 16S rRNA gene sequencing and phylogenetic analysis revealed that both strains belonged to the genus Bacillus but clearly diverged from known Bacillus species. Strain SMC 4352-1T and strain SMC 4352-2T were found to be closely related to Bacillus firmus NCIMB 9366T (98.2% sequence similarity) and Bacillus cibi JG-30T (97.1% sequence similarity), respectively. They also displayed low DNA-DNA reassociation values (less than 40%) with respect to the most closely related Bacillus species. On the basis of their polyphasic characteristics, strain SMC 4352-1T and strain SMC 4352-2T represent two novel species of the genus Bacillus, for which the names Bacillus infantis sp. nov. (type strain SMC 4352-1T=KCCM 90025T=JCM 13438T) and Bacillus idriensis sp. nov. (type strain SMC 4352-2T=KCCM 90024T=JCM 13437T) are proposed. PMID- 17082388 TI - Tuberibacillus calidus gen. nov., sp. nov., isolated from a compost pile and reclassification of Bacillus naganoensis Tomimura et al. 1990 as Pullulanibacillus naganoensis gen. nov., comb. nov. and Bacillus laevolacticus Andersch et al. 1994 as Sporolactobacillus laevolacticus comb. nov. AB - Two thermophilic strains, designated 607T and 606b, were isolated from a compost pile in Japan. The novel strains were Gram-positive, aerobic, spore-forming rods. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strains 607T and 606b were closely related to Bacillus naganoensis (94.0-94.1% similarity) and separated from clusters of the related genera Bacillus (<91.9%) and Sporolactobacillus (91.0-92.5%). In addition, some chemotaxonomic and physiological characteristics of strains 607T and 606b differed from those of B. naganoensis and the two related genera. Several differences in physiological characteristics and 16S-23S rRNA gene internal transcribed spacer region nucleotide sequences were observed between strains 607T and 606b; however, DNA DNA hybridization indicated that these two strains belonged to the same species. From these results, it is proposed that strains 607T and 606b represent the type species of a new genus, Tuberibacillus calidus gen. nov., sp. nov., with strain 607T (=JCM 13397T=DSM 17572T) as the type strain. In addition, the results of phylogenetic analyses, as well as chemotaxonomic and physiological characterization, indicated that B. naganoensis and Bacillus laevolacticus did not belong to the genus Bacillus. Based on these results, it is proposed that B. naganoensis and B. laevolacticus should be transferred to Pullulanibacillus naganoensis gen. nov., comb. nov. and Sporolactobacillus laevolacticus comb. nov., respectively. PMID- 17082389 TI - Confirmation of Thiomonas delicata (formerly Thiobacillus delicatus) as a distinct species of the genus Thiomonas Moreira and Amils 1997 with comments on some species currently assigned to the genus. AB - The transfer of Thiobacillus delicatus to the genus Thiomonas as a distinct species, Thiomonas delicata (type strain NBRC 14566T), is confirmed by its morphological and physiological properties, DNA-DNA hybridization and the grouping of its 16S rRNA gene sequence with those of other species of the genus. An emended formal description of Thiomonas delicata is given. The status of Thiomonas cuprina DSM 5495T as a member of the genus is reconsidered. PMID- 17082390 TI - Halorubrum orientale sp. nov., a halophilic archaeon isolated from Lake Ejinor, Inner Mongolia, China. AB - A motile, pleomorphic, red-pigmented archaeon, strain EJ-52T, was isolated from water from Lake Ejinor, a saline lake in Inner Mongolia, China. Analysis of the almost-complete 16S rRNA gene sequence showed that the isolate was phylogenetically related to species of the genus Halorubrum, being most closely related to Halorubrum saccharovorum ATCC 29252T (96.1% sequence similarity), Halorubrum lacusprofundi JCM 8891T (95.9%), Halorubrum tibetense AS 1.3239T (95.2%), Halorubrum alcaliphilum AS 1.3528T (95.2%) and Halorubrum vacuolatum JCM 9060T (95.1%). The polar lipids of strain EJ-52T were C20C20 derivatives of phosphatidylglycerol phosphate and phosphatidylglycerol phosphate methyl ester and a sulfated diglycosyl diether. Strain EJ-52T requires at least 2.5 M NaCl for growth and grows optimally at 3.4 M NaCl. The strain grows at 25-50 degrees C, with optimal growth occurring at 35-45 degrees C. Mg2+ is not required. The DNA G+C content is 64.2 mol%. On the basis of the data obtained in this study, strain EJ52T represents a novel species, for which the name Halorubrum orientale sp. nov. is proposed. The type strain is EJ-52T (=CECT 7145T=JCM 13889T=CGMCC 1.6295T). PMID- 17082391 TI - Pedobacter ginsengisoli sp. nov., a DNase-producing bacterium isolated from soil of a ginseng field in South Korea. AB - A Gram-negative, strictly aerobic, rod-shaped, non-motile, non-spore-forming bacterial strain, designated Gsoil 104T, was isolated from a soil sample from a ginseng field in Pocheon Province (South Korea) and was characterized taxonomically by using a polyphasic approach. On the basis of 16S rRNA gene sequence similarities, strain Gsoil 104T was shown to belong to the family Sphingobacteriaceae, being related to Pedobacter africanus DSM 12126T (97.0%), Pedobacter caeni LMG 22862T (96.9%), Pedobacter cryoconitis DSM 14825T (96.8%) and Pedobacter heparinus DSM 2366T (96.6%). The phylogenetic distance from any other Pedobacter species with a validly published name was greater than 3.4% (i.e.<96.6% 16S rRNA gene sequence similarity). DNA-DNA hybridization experiments showed that values for DNA-DNA relatedness between strain Gsoil 104T and its phylogenetically closest neighbours were below 37%. The G+C content of the genomic DNA was 43.6 mol%. The predominant respiratory quinone was MK-7. The major fatty acids were C16:1omega7c, iso-C15:0, C16:0, iso-C17:0 3-OH and iso C15:0 2-OH. These chemotaxonomic data support the affiliation of strain Gsoil 104T to the genus Pedobacter. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain Gsoil 104T represents a novel species in the genus Pedobacter, for which the name Pedobacter ginsengisoli sp. nov. is proposed. The type strain is Gsoil 104T (=KCTC 12576T=LMG 23399T). PMID- 17082392 TI - Bacillus foraminis sp. nov., isolated from a non-saline alkaline groundwater. AB - A low-G+C-content Gram-positive bacterium, designated CV53T, phylogenetically related to species of the genus Bacillus, was isolated from a highly alkaline non saline groundwater environment (pH 11.4). This organism comprised rod-shaped cells, was aerobic, did not display spore formation, was catalase- and oxidase negative, had an optimum growth temperature of 40 degrees C and had an optimum pH of approximately 7.0-8.5. Optimal growth was observed in the absence of NaCl, but growth did occur at NaCl concentrations up to 3.0%. The strain possessed an A1gamma-type peptidoglycan cell wall and the major respiratory quinone was MK-7. The predominant fatty acids were anteiso-C15:0, iso-C15:0 and anteiso-C17:0. The G+C content of the DNA was 43.1 mol%. Phylogenetic analyses of the 16S rRNA gene sequence revealed that the novel isolate is closely related to the type strain of Bacillus jeotgali, forming a coherent cluster supported by bootstrap analysis at a confidence level of 90%. The pairwise similarity of the 16S rRNA gene sequences of the two strains is 97.7%. On the basis of the phylogenetic analyses and the distinct phenotypic characteristics, strain CV53T represents a novel species within the genus Bacillus, for which we propose the name Bacillus foraminis sp. nov. The type strain is CV53T (=LMG 23174T=CIP 108889T). PMID- 17082393 TI - Mycobacterium monacense sp. nov. AB - Four bacterial strains were isolated from independent clinical specimens in different countries and their genotypic and phenotypic characters support their classification in a novel species within the genus Mycobacterium. One strain was clearly responsible for a severe, post-traumatic wound infection in a healthy boy. The novel species, for which the name Mycobacterium monacense sp. nov. is proposed, is yellow-pigmented, non-photochromogenic and grows in less than a week on solid medium. Based on phenotypic investigations alone, distinction of these four strains from known scotochromogenic rapidly growing strains is problematic. However, the novel strains differ from any other mycobacterium in each of the molecular species markers investigated: the 16S rRNA gene, the 16S-23S rRNA gene internal transcribed spacer and the hsp65 gene. Of the strains investigated, two different sequevars were detected for the hsp65 region. Phylogenetic analysis revealed that these four strains were most closely related to Mycobacterium doricum. The type strain of Mycobacterium monacense sp. nov. is B9-21-178T (=DSM 44395T=CIP 109237T). PMID- 17082394 TI - Paenibacillus xinjiangensis sp. nov., isolated from Xinjiang province in China. AB - Strain B538T is a Gram-positive, motile, rod-shaped bacterium, which was isolated from Xinjiang province in China. This organism grew optimally at 30-35 degrees C and pH 8.0-8.5. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain B538T belonged to the genus Paenibacillus and chemotaxonomic data (DNA G+C content, 47.0 mol%; major isoprenoid quinone, MK-7; cell wall type, A1gamma meso-diaminopimelic acid; major fatty acids, anteiso-C15:0 and C16:0) supported affiliation of the isolate with the genus Paenibacillus. Comparative 16S rRNA gene sequence analyses showed that the isolate was most closely related to Paenibacillus glycanilyticus DS-1T, with 16S rRNA gene sequence similarity of 98.1%; sequence similarities to other members of the genus Paenibacillus used in the phylogenetic tree were less than 96.5%. The DNA-DNA relatedness between strain B538T and P. glycanilyticus DS-1T was about 8.0%. On the basis of physiological and molecular properties, strain B538T (=KCTC 3952T=DSM 16970T) is proposed as the type strain of a novel species within the genus Paenibacillus, for which the name Paenibacillus xinjiangensis sp. nov. is proposed. PMID- 17082395 TI - Aurantimonas altamirensis sp. nov., a member of the order Rhizobiales isolated from Altamira Cave. AB - A bacterial strain, S21BT, was isolated from Altamira Cave (Cantabria, Spain). The cells were Gram-negative, short rods growing aerobically. Comparative 16S rRNA gene sequence analysis revealed that strain S21BT represented a separate subline of descent within the family 'Aurantimonadaceae' (showing 96% sequence similarity to Aurantimonas coralicida) in the order Rhizobiales (Alphaproteobacteria). The major fatty acids detected were C16:0 and C18:1omega7c. The G+C content of the DNA from strain S21BT was 71.8 mol%. Oxidase and catalase activities were present. Strain S21BT utilized a wide range of substrates for growth. On the basis of the results of this polyphasic study, isolate S21BT represents a novel species of the genus Aurantimonas, for which the name Aurantimonas altamirensis sp. nov. is proposed. The type strain is S21BT (=CECT 7138T=LMG 23375T). PMID- 17082396 TI - Nesterenkonia jeotgali sp. nov., isolated from jeotgal, a traditional Korean fermented seafood. AB - A Gram-positive, non-motile, slightly halophilic actinomycete, strain JG-241T, was isolated from jeotgal, a traditional Korean fermented seafood, and its taxonomic position was investigated by using a polyphasic approach. Strain JG 241T grew optimally at 25-30 degrees C and in the presence of 2-5% (w/v) NaCl. The physiological and biochemical properties of strain JG-241T were distinguishable from those of recognized Nesterenkonia species. Strain JG-241T had a peptidoglycan type based on L-lys-gly-D-Asp. It contained MK-7, MK-8 and MK 9 as the predominant menaquinones and anteiso-C15:0) and anteiso-C17:0 as the major fatty acids. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and an unidentified glycolipid. The DNA G+C content was 68.0 mol%. Comparative 16S rRNA gene sequence analysis showed that strain JG-241T falls within the radiation of the cluster comprising Nesterenkonia species. Similarity values between the 16S rRNA gene sequence of strain JG-241T and those of the type strains of Nesterenkonia species ranged from 96.7 to 99.7%. DNA-DNA relatedness data and repetitive extragenic palindromic DNA PCR genomic fingerprinting patterns showed that strain JG-241T differs genetically from recognized Nesterenkonia species. On the basis of its phenotypic properties and phylogenetic and genetic distinctiveness, strain JG-241T represents a novel species of the genus Nesterenkonia, for which the name Nesterenkonia jeotgali sp. nov. is proposed. The type strain is JG-241T (=KCTC 19053T=JCM 12610T). PMID- 17082397 TI - Pseudomonas segetis sp. nov., isolated from soil. AB - A Gram-negative, aerobic bacterium, designated strain FR1439T, was isolated from the soil of Dokdo in the Republic of Korea. The cells of strain FR1439T were catalase- and oxidase-positive, motile and rod-shaped. Phylogenetic analysis of the 16S rRNA gene sequence revealed that it represents a distinct line of descent within the genus Pseudomonas. The levels of DNA-DNA relatedness between strain FR1439T and type strains of phylogenetically related species, namely Pseudomonas flavescens, Pseudomonas mendocina, Pseudomonas pseudoalcaligenes and Pseudomonas straminea, ranged from 28 to 37%. Several phenotypic characteristics, together with the cellular fatty acid composition, can be used to differentiate strain FR1439T from related pseudomonads. On the basis of the polyphasic taxonomic evidence presented in this study, strain FR1439T represents a novel species, for which the name Pseudomonas segetis sp. nov. is proposed. The type strain is FR1439T (=IMSNU 14101T=CIP 108523T=KCTC 12331T). PMID- 17082398 TI - Gordonia soli sp. nov., a novel actinomycete isolated from soil. AB - A soil isolate, strain CC-AB07T, was characterized using a polyphasic approach. This organism had chemotaxonomic and morphological properties consistent with its classification in the genus Gordonia. 16S rRNA gene sequence analysis showed that the novel strain formed a monophyletic branch at the periphery of the evolutionary radiation occupied by the genus Gordonia, its closest neighbours being the type strains of Gordonia alkanivorans, Gordonia amicalis, Gordonia bronchialis, Gordonia desulfuricans, Gordonia polyisoprenivorans and Gordonia rhizosphera. The novel isolate was distinguished from all of these type strains using a range of phenotypic properties and by gyrB gene sequence analysis. It was evident from the genotypic and phenotypic data that strain CC-AB07T should be classified as representing a novel species in the genus Gordonia, for which the name Gordonia soli sp. nov. is proposed. The type strain is CC-AB07T (=BCRC 16810T=DSM 44995T). PMID- 17082399 TI - Desulfotomaculum thermosubterraneum sp. nov., a thermophilic sulfate-reducer isolated from an underground mine located in a geothermally active area. AB - A thermophilic, Gram-positive, endospore-forming, sulfate-reducing bacterium was isolated from an underground mine in a geothermally active area in Japan. Cells of this strain, designated RL50JIIIT, were rod-shaped and motile. The temperature range for growth was 50-72 degrees C (optimum growth at 61-66 degrees C) and the pH range was 6.4-7.8 (optimum at pH 7.2-7.4). Strain RL50JIIIT tolerated up to 1.5% NaCl, but optimum growth occurred in the presence of 0-1% NaCl. Electron acceptors utilized were sulfate, sulfite, thiosulfate and elemental sulfur. Electron donors utilized were H2 in the presence of CO2, alanine, various carboxylic acids and alcohols. Fermentative growth occurred on lactate and pyruvate. The cell wall contained mesodiaminopimelic acid and the major respiratory isoprenoid quinone was menaquinone 7 (MK-7). Major whole-cell fatty acids were iso-C15:0, iso-C17:0 DMA (dimethyl acetal), iso-C15:0 DMA and iso C17:0. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed 98.7% similarity with Desulfotomaculum solfataricum DSM 14956T. However, DNA-DNA hybridization experiments with Desulfotomaculum kuznetsovii, Desulfotomaculum luciae and D. solfataricum and the G+C content of the DNA (54.4 mol%) allowed the differentiation of strain RL50JIIIT from the recognized species of the genus Desulfotomaculum. Strain RL50JIIIT therefore represents a novel species, for which the name Desulfotomaculum thermosubterraneum sp. nov. is proposed. The type strain is RL50JIIIT (=DSM 16057T=JCM 13837T). PMID- 17082400 TI - Granulibacter bethesdensis gen. nov., sp. nov., a distinctive pathogenic acetic acid bacterium in the family Acetobacteraceae. AB - A Gram-negative, aerobic, coccobacillus to rod-shaped bacterium was isolated from three patients with chronic granulomatous disease. The organism was subjected to a polyphasic taxonomic study. A multilocus phylogenetic analysis based on the 16S rRNA gene, the internal transcribed spacer (ITS) region and the RecA protein demonstrated that the organism belongs to a new sublineage within the acetic acid bacteria in the family Acetobacteraceae. Phenotypic features are summarized as follows: the organism grew at an optimum temperature of 35-37 degrees C and optimum pH of 5.0-6.5. It produced a yellow pigment, oxidized lactate and acetate, the latter weakly, produced little acetic acid from ethanol and could use methanol as a sole carbon source. The two major fatty acids were a straight chain unsaturated acid (C18:1omega7c) and C16:0. The DNA base composition was 59.1 mol% G+C. The very weak production of acetic acid from ethanol, the ability to use methanol, the yellow pigmentation and high optimum temperature for growth distinguished this organism from other acetic acid bacteria. The unique phylogenetic and phenotypic characteristics suggest that the bacterium should be classified within a separate genus, for which the name Granulibacter bethesdensis gen. nov., sp. nov. is proposed. The type strain is CGDNIH1T (=ATCC BAA-1260T=DSM 17861T). PMID- 17082401 TI - Flavobacterium indicum sp. nov., isolated from warm spring water in Assam, India. AB - A polyphasic taxonomic approach was employed to characterize a strain designated GPTSA100-9T, which was isolated from water sampled from a warm spring. The micro organism, comprising Gram-negative, strictly aerobic rods, could not grow on nutritionally rich media such as tryptic soy broth. Analysis of the 16S rRNA gene sequence (1396 nt) of strain GPTSA100-9T revealed that it is a member of the genus Flavobacterium, sharing 99.8% sequence similarity with the CFB group bacterium strain A0653 (AF236016), 93.4% with '[Flexibacter] aurantiacus subsp. excathedrus' and 93.2-92.0% with Flavobacterium saliperosum, Flavobacterium soli, Flavobacterium aquatile and Flavobacterium columnare. The G+C content of the genomic DNA was 31.0 mol%. The major fatty acids of the strain grown on modified R2A agar were iso-C15:0 (18.5%), iso-C15:1 G (18.0%), summed feature 3 (iso-C15:0 2-OH and/or C16:1omega7c, 16.6%) and iso-C17:0 3-OH (9.0%). On the basis of phenotypic and genotypic characteristics, strain GPTSA100-9T represents a novel species of the genus Flavobacterium, for which the name Flavobacterium indicum sp. nov. is proposed. The type strain is GPTSA100-9T (=MTCC 6936T=DSM 17447T). PMID- 17082402 TI - Anaerovirgula multivorans gen. nov., sp. nov., a novel spore-forming, alkaliphilic anaerobe isolated from Owens Lake, California, USA. AB - A novel, alkaliphilic, obligately anaerobic bacterium, strain SCAT, was isolated from mud sediments of a soda lake in California, USA. The rod-shaped cells were motile, Gram-positive, formed spores and were 0.4-0.5x2.5-5.0 microm in size. Growth occurred within the pH range 6.7-10.0 and was optimal at pH 8.5. The temperature range for growth was 10-45 degrees C, with optimal growth at 35 degrees C. NaCl was required for growth. Growth occurred at 0.5-9.0% (w/v) NaCl and was optimal at 1-2% (w/v). The novel isolate was a catalase-negative chemo organoheterotroph that fermented sugars, proteolysis products, some organic and amino acids, glycerol, d-cellobiose and cellulose. It was also capable of growth by the Stickland reaction. Strain SCAT was sensitive to tetracycline, chloramphenicol, rifampicin and gentamicin, but it was resistant to ampicillin and kanamycin. The G+C content of the genomic DNA was 34.2 mol%. Major fatty acid components were C14:0, iso-C15:0, C16:1omega9c and C16:0. 16S rRNA gene sequence analysis of strain SCAT showed a similarity of approximately 97% with the type strains of Clostridium formicaceticum and Clostridium aceticum in clostridial cluster XI and a similarity of less than 94.2% to any other recognized Clostridium species and those of related genera in this cluster. Strain SCAT was clearly differentiated from C. formicaceticum and C. aceticum based on comparison of their phenotypic properties and fatty acid profiles, as well as low levels of DNA-DNA relatedness between strain SCAT and the type strains of these two species. Therefore, strain SCAT is considered to represent a novel species of a new genus, Anaerovirgula multivorans gen. nov., sp. nov., in clostridial cluster XI. The type strain is SCAT (=ATCC BAA-1084T=JCM 12857T=DSM 17722T=CIP 107910T). PMID- 17082403 TI - Rhizobium lusitanum sp. nov. a bacterium that nodulates Phaseolus vulgaris. AB - The species Phaseolus vulgaris is a promiscuous legume nodulated by several species of the family Rhizobiaceae. During a study of rhizobia nodulating this legume in Portugal, we isolated several strains that nodulate P. vulgaris effectively and also Macroptilium atropurpureum and Leucaena leucocephala, but they form ineffective nodules in Medicago sativa. According to phylogenetic analysis of the 16S rRNA gene sequence, the strains from this study belong to the genus Rhizobium, with Rhizobium rhizogenes and Rhizobium tropici as the closest related species, with 99.9 and 99.2% similarity, respectively, between the type strains of these species and strain P1-7T. The nodD and nifH genes carried by strain P1-7T are phylogenetically related to those of other species nodulating Phaseolus. This strain does not carry virulence genes present in the type strain of R. rhizogenes, ATCC 11325T. Analysis of the recA and atpD genes confirms this phylogenetic arrangement, showing low similarity with respect to those of R. rhizogenes ATCC 11325T (91.9 and 94.1% similarity, respectively) and R. tropici IIB CIAT 899T (90.6% and 91.8% similarity, respectively). The intergenic spacer (ITS) of the strains from this study is phylogenetically divergent from those of R. rhizogenes ATCC 11235T and R. tropici CIAT 899T, with 85.9 and 82.8% similarity, respectively, with respect to strain P1-7T. The tRNA profile and two primer random amplified polymorphic DNA pattern of strain P1-7T are also different from those of R. rhizogenes ATCC 11235T and R. tropici CIAT 899T. The strains isolated in this study can be also differentiated from R. rhizogenes and R. tropici by several phenotypic characteristics. The results of DNA-DNA hybridization showed means of 28 and 25% similarity between strain P1-7T and R. rhizogenes ATCC 11235T and R. tropici CIAT 899T, respectively. All these data showed that the strains isolated in this study belong to a novel species of the genus Rhizobium, for which we propose the name Rhizobium lusitanum sp. nov.; the type strain is P1-7T (=LMG 22705T=CECT 7016T). PMID- 17082404 TI - Ferrimonas futtsuensis sp. nov. and Ferrimonas kyonanensis sp. nov., selenate reducing bacteria belonging to the Gammaproteobacteria isolated from Tokyo Bay. AB - Two novel mesophilic, facultatively anaerobic, selenate-reducing bacteria, designated strains FUT3661T and Asr22-7T, were isolated from a sediment sample and the alimentary tract of littleneck clams, respectively. Both sources of the samples were collected from the coast of Tokyo Bay, Japan. Cells were Gram negative rods and motile by means of a polar flagellum. The strains reduced selenate to elemental selenium (Se0) and also reduced iron(III) oxyhydroxide, iron(III) citrate, arsenate, manganese(IV) oxide, elemental sulfur and oxygen and used lactate, pyruvate, yeast extract, tryptone and Casamino acids as electron donors and carbon sources. The strains contained both menaquinone (MK-7) and ubiquinones (Q-7 and Q-8) as isoprenoid quinones. The major fatty acids were C16:0 and C16:1omega9c. The G+C content of the genomic DNA was 58.1 mol% for strain FUT3661T and 57.2 mol% for strain Asr22-7T. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the strains were related to members of the genus Ferrimonas (<94.0% similarities), although the two novel strains formed a separate lineage. 16S rRNA gene sequence similarity between strains FUT3661T and Asr22-7T was 96%. On the basis of this polyphasic analysis, it was concluded that strains FUT3661T and Asr22-7T represent two novel species within the genus Ferrimonas, for which the names Ferrimonas futtsuensis sp. nov. (type strain FUT3661T=NBRC 101558T=DSM 18154T) and Ferrimonas kyonanensis sp. nov. (type strain Asr22-7T=NBRC 101286T=DSM 18153T) are proposed. PMID- 17082405 TI - Salicola salis sp. nov., an extremely halophilic bacterium isolated from Ezzemoul sabkha in Algeria. AB - A novel, extremely halophilic bacterium was isolated from brine samples collected from Ezzemoul sabkha in north-east Algeria. Cells of this isolate, designated B2T, were Gram-negative, rod-shaped and motile. Growth occurred between 10 and 25% (w/v) NaCl and the isolate grew optimally at 15-20% (w/v) NaCl. The pH range for growth was 6.0-9.0 with an optimum at pH 7.0-7.5. The predominant fatty acids were C16:0 and C18:1omega9c. Other fatty acids present were C16:1omega9c, C18:0 10-methyl, C12:0 3-OH, C10:0 and C12:0. The G+C content of the genomic DNA was 56.0 mol%. 16S rRNA gene sequence analysis indicated that strain B2T was closely related to Salicola marasensis in the Gammaproteobacteria. The level of 16S rRNA gene sequence similarity between strain B2T and the type strain of Salicola marasensis was 99%. DNA-DNA hybridization experiments between strain B2T and Salicola marasensis indicated a level of relatedness of 52%. The phenotypic characteristics of strain B2T allowed its differentiation from recognized species of the genus Salicola. Strain B2T was able to hydrolyse starch but not aesculin. It was unable to use carbohydrates and could not use citrate, pyruvate or succinate as sole carbon and energy sources. On the basis of the polyphasic data presented, strain B2T is considered to represent a novel species of the genus Salicola, for which the name Salicola salis sp. nov. is proposed. The type strain is B2T (=CECT 7106T=LMG 23122T). PMID- 17082406 TI - Marinobacter koreensis sp. nov., isolated from sea sand in Korea. AB - A marine, Gram-negative, aerobic, motile, straight-rod-shaped, moderately halophilic bacterium, designated strain DD-M3T, was isolated from sea sand in Pohang, Korea. A phylogenetic tree based on 16S rRNA gene sequences showed that the strain fell within the evolutionary radiation encompassed by the genus Marinobacter. The levels of 16S rRNA gene sequence similarity between the novel strain and the type strains of recognized Marinobacter species ranged from 94.2 to 97.6%, the highest values being with Marinobacter flavimaris SW-145T (97.6%) and Marinobacter lipolyticus SM19T (96.8%). The values for DNA-DNA relatedness between isolate DD-M3T and the type strains of the most closely related species, M. flavimaris and M. lipolyticus, were 41 and 36%, respectively. Strain DD-M3T was characterized as having Q-9 as the predominant respiratory quinone and 16:0, summed feature 3 and 18:1omega9c as the main fatty acids. The DNA G+C content was 54.1 mol%. On the basis of its phenotypic and genotypic characteristics, DD-M3T represents a novel species of the genus Marinobacter, for which the name Marinobacter koreensis sp. nov. is proposed, with DD-M3T (=KACC 11513T=DSM 17924T) as the type strain. PMID- 17082407 TI - Pseudomonas moraviensis sp. nov. and Pseudomonas vranovensis sp. nov., soil bacteria isolated on nitroaromatic compounds, and emended description of Pseudomonas asplenii. AB - Two strains of Gram-negative bacteria isolated from soil by selective enrichment with nitroaromatics were subjected to a polyphasic taxonomic study. On the basis of 16S rRNA gene sequence analysis, the two strains were found to belong to the genus Pseudomonas, within the Gammaproteobacteria. Strain 1B4T shared the highest sequence similarity with Pseudomonas koreensis DSM 16610T (99.5%) and Pseudomonas jessenii CCM 4840T (99.3%), and strain 2B2T with Pseudomonas asplenii DSM 17133T (98.9%), Pseudomonas fuscovaginae DSM 7231T (98.9%) and Pseudomonas putida DSM 291T (98.7%). On the basis of phylogenetic analysis, DNA-DNA hybridization and phenotype, including chemotaxonomic characteristics, two novel species, Pseudomonas moraviensis sp. nov. with the type strain 1B4T (=CCM 7280T=DSM 16007T) and Pseudomonas vranovensis sp. nov. with the type strain 2B2T (=CCM 7279T=DSM 16006T), are proposed. The description of P. asplenii was emended on the basis of additional data obtained in this study. PMID- 17082408 TI - Brevibacillus ginsengisoli sp. nov., a denitrifying bacterium isolated from soil of a ginseng field. AB - A Gram-positive, rod-shaped, spore-forming bacterium, Gsoil 3088T, was isolated from soil from a ginseng field in Pocheon Province in South Korea and characterized in order to determine its taxonomic position. On the basis of 16S rRNA gene sequence similarity, strain Gsoil 3088T was shown to belong to the family Paenibacillaceae, being related to Brevibacillus centrosporus (96.6%), Brevibacillus borstelensis (96.3%), Brevibacillus parabrevis (96.1%), Brevibacillus formosus (96.1%), Brevibacillus brevis (96.1%) and Brevibacillus laterosporus (96.0%). The phylogenetic distances from other validly described species within the genus Brevibacillus were greater than 4.0% (i.e. there was less than 96.0% similarity). The G+C content of the genomic DNA was 52.1 mol%. Phenotypic and chemotaxonomic data (major menaquinone, MK-7; fatty acid profile, iso-C15:0, iso-C14:0 and anteiso-C15:0) supported the affiliation of strain Gsoil 3088T to the genus Brevibacillus. The results of physiological and biochemical tests allowed strain Gsoil 3088T to be distinguished genotypically and phenotypically from Brevibacillus species with validly published names. Strain Gsoil 3088T, therefore, represents a novel species of the genus Brevibacillus, for which the name Brevibacillus ginsengisoli sp. nov. is proposed. The type strain is Gsoil 3088T (=KCTC 13938T=LMG 23403T). PMID- 17082409 TI - Pseudomonas simiae sp. nov., isolated from clinical specimens from monkeys (Callithrix geoffroyi). AB - An unusual Gram-negative, catalase- and oxidase-positive, rod-shaped bacterium isolated from different clinical samples from two monkeys (Callithrix geoffroyi) was characterized by phenotypic and molecular genetic methods. The micro-organism was tentatively identified as a Pseudomonas species on the basis of the results of cellular morphological and biochemical tests. Fatty acid studies confirmed this generic placement and comparative 16S rRNA gene sequencing studies demonstrated that the unknown isolates were phylogenetically closely related to each other (100% sequence similarity) and were part of the 'Pseudomonas fluorescens intrageneric cluster'. The novel bacterium, however, was distinguished from other phylogenetically related species of Pseudomonas by DNA DNA hybridization studies and biochemical tests. On the basis of both phenotypic and phylogenetic findings, it is proposed that the novel Pseudomonas isolates are classified as Pseudomonas simiae sp. nov. The type strain of P. simiae is OLiT (=CCUG 50988T=CECT 7078T). PMID- 17082410 TI - Paenibacillus panacisoli sp. nov., a xylanolytic bacterium isolated from soil in a ginseng field in South Korea. AB - A Gram-positive, facultatively anaerobic, motile, spore-forming bacterium, designated Gsoil 1411T, was isolated from soil of a ginseng field in Pocheon Province (South Korea) and was characterized using a polyphasic approach. Comparative analysis of 16S rRNA gene sequences revealed that strain Gsoil 1411T belongs to the family Paenibacillaceae, with closest sequence similarity to the type strains of Paenibacillus xylanilyticus (95.7%), Paenibacillus illinoisensis (95.2%) and Paenibacillus pabuli (94.8%). Strain Gsoil 1411T showed less than 94% sequence similarity to the type strains of other recognized members of the genus Paenibacillus. In addition, the presence of MK-7 as the major menaquinone, anteiso-C15:0 as a major fatty acid (44.8%) and the presence of PAEN513F and PAEN862F signature sequences suggest that it is affiliated to the genus Paenibacillus. The G+C content of the genomic DNA was 53.9 mol%. On the basis of its phenotypic characteristics and phylogenetic distinctiveness, strain Gsoil 1411T is suggested to represent a novel species within the genus Paenibacillus, for which the name Paenibacillus panacisoli sp. nov. is proposed. The type strain is Gsoil 1411T (=KCTC 13020T=LMG 23405T). PMID- 17082411 TI - Natrinema ejinorense sp. nov., isolated from a saline lake in Inner Mongolia, China. AB - A Gram-negative, non-motile, neutrophilic, pleomorphic and extremely halophilic archaeon, strain EJ-57T, was isolated from saline Lake Ejinor in Inner Mongolia, China. Strain EJ-57T was able to grow at 25-50 degrees C, required at least 1.8 M NaCl for growth (optimum at 3.4 M NaCl) and grew over a pH range from 6.0 to 8.5 (optimum at pH 7.0). Hypotonic treatment with less than 1.5 M NaCl caused cell lysis. Analysis of the almost complete 16S rRNA gene sequence indicated that the isolate represented a member of the genus Natrinema in the family Halobacteriaceae. Strain EJ-57T was most closely related to Natrinema versiforme JCM 10478T (96.2% sequence similarity), Natrinema pallidum NCIMB 777T (95.9% sequence similarity), Natrinema altunense JCM 12890T (95.8% sequence similarity) and Natrinema pellirubrum NCIMB 786T (95.5 % sequence similarity). However, DNA DNA hybridization experiments showed that strain EJ-57T was not related to these species, with levels of DNA-DNA relatedness equal to or below 39%. The major polar lipids of the isolate were C20C20 and C20C25 derivatives of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and the disulfated glycolipid S2-DGA-1. The G+C content of the genomic DNA was 64.7 mol%. Comparative analysis of phenotypic characteristics between strain EJ-57T and recognized Natrinema species supported the conclusion that EJ-57T represents a novel species within this genus, for which the name Natrinema ejinorense sp. nov. is proposed. The type strain is EJ-57T (=CECT 7144T=JCM 13890T=CGMCC 1.6202T). PMID- 17082412 TI - Devosia soli sp. nov., isolated from greenhouse soil in Korea. AB - A Gram-negative, obligately aerobic, rod-shaped bacterium was isolated from greenhouse soil used to cultivate lettuce. The strain, GH2-10T, was characterized on the basis of phenotypic and genotypic data. 16S rRNA gene sequence analysis revealed that the isolate belonged to the genus Devosia, with highest sequence similarity (98.5%) to Devosia riboflavina IFO 13584T. Sequence similarities with other strains tested were below 97.0%. Strain GH2-10T had Q-10 as the predominant ubiquinone and C18:1omega7c and C16:0 as the major fatty acids. The G+C content of the genomic DNA was 59.5 mol%. The results of DNA-DNA hybridization experiments (47% relatedness between D. riboflavina DSM 7230T and strain GH2-10T) and physiological and biochemical tests suggested that strain GH2-10T represents a novel species of the genus Devosia, for which the name Devosia soli sp. nov. is proposed. The type strain is GH2-10T (=KACC 11509T=DSM 17780T). PMID- 17082413 TI - Paracoccus sulfuroxidans sp. nov., a sulfur oxidizer from activated sludge. AB - Bacterial strain LW36T was isolated from activated sludge of a wastewater treatment bioreactor. Cells of strain LW36T were Gram-negative coccoids to short rods, 1.0-1.2x0.5 microm in size. Colonies were cream-coloured, smooth and circular. Strain LW36T was hetero-organotrophic and chemolithotrophic and was able to use reduced sulfur as an energy resource. Growth was observed at 25-36 degrees C and pH 5-10. The most abundant cellular fatty acid of strain LW36T was C18:1omega7c (64.2%). The sole respiratory quinone was ubiquinone-10. The G+C content of the genomic DNA was 61.3 mol% (Tm). 16S rRNA gene sequence analysis indicated that strain LW36T was phylogenetically related to members of the genus Paracoccus, with similarities ranging from 92.4 to 94.9%. Based on these results, it is concluded that strain LW36T represents a novel species of the genus Paracoccus, for which the name Paracoccus sulfuroxidans is proposed. The type strain is strain LW36T (=CGMCC 1.5364T=JCM 14013T). PMID- 17082414 TI - Exophiala crusticola anam. nov. (affinity Herpotrichiellaceae), a novel black yeast from biological soil crusts in the Western United States. AB - A novel black yeast-like fungus, Exophiala crusticola, is described based on two closely related isolates from biological soil crust (BSC) samples collected on the Colorado Plateau (Utah) and in the Great Basin desert (Oregon), USA. Their morphology places them in the anamorphic genus Exophiala, having affinities to the family Herpotrichiellaceae (Ascomycota). Phylogenetic analysis of their D1/D2 large subunit nuclear ribosomal RNA (LSU nrRNA) gene sequences suggests that they represent a distinct species. The closest known putative relative to Exophiala crusticola is Capronia coronata Samuels, isolated from decorticated wood in Westland County, New Zealand. The holotype for Exophiala crusticola anam. nov. is UAMH 10686 and the type strain is CP141bT (=ATCC MYA-3639T=CBS 119970T=DSM 16793T). Dark-pigmented fungi appear to constitute an important heterotrophic component of soil crusts and Exophiala crusticola represents the first description of a dematiaceous fungus isolated from BSCs. PMID- 17082416 TI - Lysobacter enzymogenes subsp. enzymogenes Christensen and Cook 1978, L. enzymogenes subsp. cookii Christensen 1978 and Streptococcus casseliflavus (Mundt and Graham 1968) Vaughan et al. 1978 should have been cited in the Approved Lists of Bacterial Names. Request for an opinion. AB - Lysobacter enzymogenes subsp. enzymogenes Christensen and Cook 1978, L. enzymogenes subsp. cookii Christensen 1978 and Streptococcus casseliflavus (Mundt and Graham 1968) Vaughan et al. 1978 were inadvertently omitted from the Approved Lists of Bacterial Names. According to Rule 24a, Note 1, the authors request that these names be considered as included in these Lists. PMID- 17082415 TI - Cryptococcus anemochoreius sp. nov., a novel anamorphic basidiomycetous yeast isolated from the atmosphere in central South Africa. AB - A novel yeast strain, CBS 10258T, was isolated from the atmosphere in central South Africa. Sequence analysis of the D1/D2 domain and internal transcribed spacer region of the novel strain indicates that it represents a novel species within the Cryptococcus laurentii complex. Phylogenetic analyses based on the D1/D2 domain revealed that the novel strain occupies a relatively isolated position within this complex with Papiliotrema bandonii, Cryptococcus perniciosus, Cryptococcus nemorosus and Cryptococcus sp. CBS 8363 being the closest relatives. However, the novel strain could be distinguished from related species by standard physiological tests including the inability to assimilate rhamnose, methyl alpha-d-glucoside, salicin, lactose, erythritol, ribitol, xylitol, citrate and ethanol. In addition, no extracellular starch production was observed and the isolate was able to grow in the absence of additional vitamins. On the basis of these results, we suggest that the new strain represents a novel species for which the name Cryptococcus anemochoreius sp. nov. is proposed [type strain CBS 10258T (=NRRL Y-27920T)]. PMID- 17082417 TI - Proposal of Parvimonas gen. nov. and Quatrionicoccus gen. nov. as replacements for the illegitimate, prokaryotic, generic names Micromonas Murdoch and Shah 2000 and Quadricoccus Maszenan et al. 2002, respectively. AB - The prokaryotic, generic names Micromonas Murdoch and Shah 2000 and Quadricoccus Maszenan et al. 2002 are illegitimate* because they are later homonyms of the names Micromonas Manton and Parke 1960 (alga) and Quadricoccus Fott 1948 (alga). [Principle 2, Rule 51b(4) of the Bacteriological Code (1990 Revision)]. Such names have no claim to be correct names (Principle 6) and, therefore, replacement generic names must be proposed (Rule 54). PMID- 17082418 TI - Valid publication of names of prokaryotes according to the rules of nomenclature: past history and current practice. AB - It is now 30 years since the concept of valid publication was introduced into prokaryote systematics in such a way that a system of registration/indexing was integrated. Despite the success of this system and the introduction of the Approved Lists of Bacterial Names, accompanied by a new starting date in prokaryote nomenclature, there is a need to review both the historical aspects and current practices so that the current generation of prokaryote systematists have an up-to-date overview of some of the key elements. PMID- 17082419 TI - Probing the chiroptical response of a single molecule. AB - Chirally sensitive measurement techniques have generally been restricted to bulk samples. Here, we report the observation of fluorescence-detected circular dichroism (FDCD) from single (bridgedtriarylamine) helicene molecules by using an excitation wavelength (457 nanometers) in the vicinity of an electronic transition that shows circular dichroism in bulk samples. The distributions of dissymmetry (g) parameters by analysis of signals from pure M- and P-type diastereomers are almost perfect mirror images of one another, each spanning a range of both positive and negative values. In addition, we observe a well defined structure in the histogram of dissymmetry parameters suggestive of specific molecular orientations at the polymer interface. These single-molecule results highlight strong intrinsic circular dichroism responses that can be obscured by cancellation effects in ensemble measurements of a randomly oriented bulk sample. PMID- 17082420 TI - Localization of iron in Arabidopsis seed requires the vacuolar membrane transporter VIT1. AB - Iron deficiency is a major human nutritional problem wherever plant-based diets are common. Using synchrotron x-ray fluorescence microtomography to directly visualize iron in Arabidopsis seeds, we show that iron is localized primarily to the provascular strands of the embryo. This localization is completely abolished when the vacuolar iron uptake transporter VIT1 is disrupted. Vacuolar iron storage is also critical for seedling development because vit1-1 seedlings grow poorly when iron is limiting. We have uncovered a fundamental aspect of seed biology that will ultimately aid the development of nutrient-rich seed, benefiting both human health and agricultural productivity. PMID- 17082421 TI - WNT and DKK determine hair follicle spacing through a reaction-diffusion mechanism. AB - Mathematical reaction-diffusion models have been suggested to describe formation of animal pigmentation patterns and distribution of epidermal appendages. However, the crucial signals and in vivo mechanisms are still elusive. Here we identify WNT and its inhibitor DKK as primary determinants of murine hair follicle spacing, using a combined experimental and computational modeling approach. Transgenic DKK overexpression reduces overall appendage density. Moderate suppression of endogenous WNT signaling forces follicles to form clusters during an otherwise normal morphogenetic program. These results confirm predictions of a WNT/DKK-specific mathematical model and provide in vivo corroboration of the reaction-diffusion mechanism for epidermal appendage formation. PMID- 17082423 TI - Salary survey. PMID- 17082422 TI - Phytoplankton and cloudiness in the Southern Ocean. AB - The effect of ocean biological productivity on marine clouds is explored over a large phytoplankton bloom in the Southern Ocean with the use of remotely sensed data. Cloud droplet number concentration over the bloom was twice what it was away from the bloom, and cloud effective radius was reduced by 30%. The resulting change in the short-wave radiative flux at the top of the atmosphere was -15 watts per square meter, comparable to the aerosol indirect effect over highly polluted regions. This observed impact of phytoplankton on clouds is attributed to changes in the size distribution and chemical composition of cloud condensation nuclei. We propose that secondary organic aerosol, formed from the oxidation of phytoplankton-produced isoprene, can affect chemical composition of marine cloud condensation nuclei and influence cloud droplet number. Model simulations support this hypothesis, indicating that 100% of the observed changes in cloud properties can be attributed to the isoprene secondary organic aerosol. PMID- 17082424 TI - NASA space science. Hubble gets a green light, with other missions on hold. PMID- 17082425 TI - U.S. politics. Scientists look to Missouri to show the way on stem cells. PMID- 17082426 TI - Research funding. Olympics-level costs upset plan to move U.K. biomedical institute. PMID- 17082427 TI - Conflict of interest. NIH rules rile scientists, survey finds. PMID- 17082428 TI - Genetics. Small RNAs reveal an activating side. PMID- 17082429 TI - Avian influenza. New H5N1 strain emerges in southern China. PMID- 17082430 TI - Bioterrorism agents. U.S. panel wants security rules applied to genomes, not pathogens. PMID- 17082431 TI - AIDS research. Center puts hold on mangabey experiments. PMID- 17082432 TI - Ecology. Global loss of biodiversity harming ocean bounty. PMID- 17082433 TI - Wildlife conservation. The carnivore comeback. PMID- 17082434 TI - Wildlife conservation. On the brink. PMID- 17082436 TI - U.S. research policy. Congress cancels contentious program to bolster industry. PMID- 17082435 TI - Materials science. Japanese latecomer joins race to build a hard x-ray laser. PMID- 17082437 TI - How protected are coral reefs? PMID- 17082438 TI - Comment on "Rapid uplift of the Altiplano revealed through 13C-18O bonds in paleosol carbonates". AB - Based on stable isotope measurements, Ghosh et al. (Reports, 27 January 2006, p. 511) concluded that the Bolivian Altiplano uplifted 3 to 4 kilometers between approximately 10.3 and approximately 6.7 million years ago as a result of gravitational loss of dense lithosphere. This result stands at odds with current geological knowledge of the Central Andes, and we propose a test for the reliability of the paleoaltimetry method. PMID- 17082439 TI - Climate change. An ambitious, centrist approach to global warming legislation. PMID- 17082440 TI - Cell biology. Nuclear pore complex models gel. PMID- 17082441 TI - Materials science. Polymers in the pore. PMID- 17082442 TI - Materials science. Colloidal encounters: a matter of attraction. PMID- 17082443 TI - Physics. Beyond the particle in the box. PMID- 17082444 TI - Ecology. A renaissance in the study of abundance. PMID- 17082445 TI - Astronomy. Radio traces of cosmic shock waves. PMID- 17082446 TI - Biomedicine. Life, the universe, and body temperature. PMID- 17082447 TI - A century of Alzheimer's disease. AB - One hundred years ago a small group of psychiatrists described the abnormal protein deposits in the brain that define the most common neurodegenerative diseases. Over the past 25 years, it has become clear that the proteins forming the deposits are central to the disease process. Amyloid-beta and tau make up the plaques and tangles of Alzheimer's disease, where these normally soluble proteins assemble into amyloid-like filaments. Tau inclusions are also found in a number of related disorders. Genetic studies have shown that dysfunction of amyloid-beta or tau is sufficient to cause dementia. The ongoing molecular dissection of the neurodegenerative pathways is expected to lead to a true understanding of disease pathogenesis. PMID- 17082448 TI - 100 years and counting: prospects for defeating Alzheimer's disease. AB - This week marks a century since the first description of Alzheimer's disease (AD). Despite approval of several drugs for AD, the disease continues to rob millions of their memories and their lives. Fortunately, many new therapies directly targeting the mechanisms underlying AD are now in the pipeline. Among the investigative AD therapies in clinical trials are several strategies to block pathogenic amyloid-beta peptides and to rescue vulnerable neurons from degeneration. Complementary but less mature strategies aim to prevent the copathogenic effects of apolipoprotein E and the microtubule-associated protein tau. New insights into selective neuronal vulnerability and the link between aging and AD may provide additional entry points for therapeutic interventions. The predicted increase in AD cases over the next few decades makes the development of better treatments a matter of utmost importance and urgency. PMID- 17082449 TI - Accelerated evolution of conserved noncoding sequences in humans. AB - Changes in gene regulation likely influenced the profound phenotypic divergence of humans from other mammals, but the extent of adaptive substitution in human regulatory sequences remains unknown. We identified 992 conserved noncoding sequences (CNSs) with a significant excess of human-specific substitutions. These accelerated elements were disproportionately found near genes involved in neuronal cell adhesion. To assess the uniqueness of human noncoding evolution, we examined CNSs accelerated in chimpanzee and mouse. Although we observed a similar enrichment near neuronal adhesion genes in chimpanzee, the accelerated CNSs themselves exhibited almost no overlap with those in human, suggesting independent evolution toward different neuronal phenotypes in each species. CNSs accelerated in mouse showed no bias toward neuronal cell adhesion. Our results indicate that widespread cis-regulatory changes in human evolution may have contributed to uniquely human features of brain development and function. PMID- 17082450 TI - Impacts of biodiversity loss on ocean ecosystem services. AB - Human-dominated marine ecosystems are experiencing accelerating loss of populations and species, with largely unknown consequences. We analyzed local experiments, long-term regional time series, and global fisheries data to test how biodiversity loss affects marine ecosystem services across temporal and spatial scales. Overall, rates of resource collapse increased and recovery potential, stability, and water quality decreased exponentially with declining diversity. Restoration of biodiversity, in contrast, increased productivity fourfold and decreased variability by 21%, on average. We conclude that marine biodiversity loss is increasingly impairing the ocean's capacity to provide food, maintain water quality, and recover from perturbations. Yet available data suggest that at this point, these trends are still reversible. PMID- 17082451 TI - Giant ringlike radio structures around galaxy cluster Abell 3376. AB - In the current paradigm of cold dark matter cosmology, large-scale structures are assembling through hierarchical clustering of matter. In this process, an important role is played by megaparsec (Mpc)-scale cosmic shock waves, arising in gravity-driven supersonic flows of intergalactic matter onto dark matter dominated collapsing structures such as pancakes, filaments, and clusters of galaxies. Here, we report Very Large Array telescope observations of giant ( approximately 2 Mpc by 1.6 Mpc), ring-shaped nonthermal radio-emitting structures, found at the outskirts of the rich cluster of galaxies Abell 3376. These structures may trace the elusive shock waves of cosmological large-scale matter flows, which are energetic enough to power them. These radio sources may also be the acceleration sites where magnetic shocks are possibly boosting cosmic ray particles with energies of up to 10(18) to 10(19) electron volts. PMID- 17082452 TI - Imaging the sublimation dynamics of colloidal crystallites. AB - We studied the kinetics of sublimating crystals with single-particle resolution by experiments with colloidal spheres and by computer simulations. A short-range attraction between spheres led to crystallites one to three layers thick. The spheres were tracked with optical microscopy while the attraction was reduced and the crystals sublimated. Large crystallites sublimated by escape of particles from the perimeter. The rate of shrinkage was greatly enhanced, however, when the size decreased to less than 20 to 50 particles, depending on the location in the phase diagram. At this size, the crystallites transformed into a dense amorphous structure, which rapidly vaporized. The enhancement of kinetics by metastable or unstable phases may play a major role in the melting, freezing, and annealing of crystals. PMID- 17082453 TI - Pattern formation and shape changes in self-oscillating polymer gels. AB - We developed an efficient model for responsive gels that captures large-scale, two-dimensional (2D) deformations and chemical reactions within a swollen polymer network. The 2D calculations allowed us to probe not only volume changes but also changes in sample shape. By focusing on gels undergoing the oscillatory Belousov Zhabotinsky reaction, we observed traveling waves of local swelling that form a rich variety of dynamic patterns and give rise to distinctive oscillations in the gel's shape. The observed patterns depend critically on the gel's dimensions. The approach provides a useful computational tool for probing the dynamics of chemomechanical processes and uncovering morphological transformations in responsive gels. PMID- 17082454 TI - Renewable hydrogen from nonvolatile fuels by reactive flash volatilization. AB - Droplets of nonvolatile fuels such as soy oil and glucose-water solutions can be flash evaporated by catalytic partial oxidation to produce hydrogen in high yields with a total time in the reactor of less than 50 milliseconds. Pyrolysis, coupled with catalytic oxidation of the fuels and their fragments upon impact with a hot rhodium-cerium catalyst surface, avoids the formation of deactivating carbon layers on the catalyst. The catalytic reactions of these products generate approximately 1 megawatt of heat per square meter, which maintains the catalyst surface above 800 degrees C at high drop impact rates. At these temperatures, heavy fuels can be catalytically transformed directly into hydrogen, carbon monoxide, and other small molecules in very short contact times without the formation of carbon. PMID- 17082455 TI - Coherent electronic fringe structure in incommensurate silver-silicon quantum wells. AB - Atomically uniform silver films grown on highly doped n-type Si(111) substrates show fine-structured electronic fringes near the silicon valence band edge as observed by angle-resolved photoemission. No such fringes are observed for silver films grown on lightly doped n-type substrates or p-type substrates, although all cases exhibited the usual quantum-well states corresponding to electron confinement in the film. The fringes correspond to electronic states extending over the silver film as a quantum well and reaching into the silicon substrate as a quantum slope, with the two parts coherently coupled through an incommensurate interface structure. PMID- 17082456 TI - FG-rich repeats of nuclear pore proteins form a three-dimensional meshwork with hydrogel-like properties. AB - Nuclear pore complexes permit rapid passage of cargoes bound to nuclear transport receptors, but otherwise suppress nucleocytoplasmic fluxes of inert macromolecules >/=30 kilodaltons. To explain this selectivity, a sieve structure of the permeability barrier has been proposed that is created through reversible cross-linking between Phe and Gly (FG)-rich nucleoporin repeats. According to this model, nuclear transport receptors overcome the size limit of the sieve and catalyze their own nuclear pore-passage by a competitive disruption of adjacent inter-repeat contacts, which transiently opens adjoining meshes. Here, we found that phenylalanine-mediated inter-repeat interactions indeed cross-link FG-repeat domains into elastic and reversible hydrogels. Furthermore, we obtained evidence that such hydrogel formation is required for viability in yeast. PMID- 17082457 TI - Protrudin induces neurite formation by directional membrane trafficking. AB - Guanosine triphosphatases of the Rab family are key regulators of membrane trafficking, with Rab11 playing a specific role in membrane recycling. We identified a mammalian protein, protrudin, that promoted neurite formation through interaction with the guanosine diphosphate (GDP)-bound form of Rab11. Phosphorylation of protrudin by extracellular signal-regulated kinase (ERK) in response to nerve growth factor promoted protrudin association with Rab11-GDP. Down-regulation of protrudin by RNA interference induced membrane extension in all directions and inhibited neurite formation. Thus, protrudin regulates Rab11 dependent membrane recycling to promote the directional membrane trafficking required for neurite formation. PMID- 17082458 TI - Where water is oxidized to dioxygen: structure of the photosynthetic Mn4Ca cluster. AB - The oxidation of water to dioxygen is catalyzed within photosystem II (PSII) by a Mn(4)Ca cluster, the structure of which remains elusive. Polarized extended x-ray absorption fine structure (EXAFS) measurements on PSII single crystals constrain the Mn(4)Ca cluster geometry to a set of three similar high-resolution structures. Combining polarized EXAFS and x-ray diffraction data, the cluster was placed within PSII, taking into account the overall trend of the electron density of the metal site and the putative ligands. The structure of the cluster from the present study is unlike either the 3.0 or 3.5 angstrom-resolution x-ray structures or other previously proposed models. PMID- 17082459 TI - Transgenic mice with a reduced core body temperature have an increased life span. AB - Reduction of core body temperature has been proposed to contribute to the increased life span and the antiaging effects conferred by calorie restriction (CR). Validation of this hypothesis has been difficult in homeotherms, primarily due to a lack of experimental models. We report that transgenic mice engineered to overexpress the uncoupling protein 2 in hypocretin neurons (Hcrt-UCP2) have elevated hypothalamic temperature. The effects of local temperature elevation on the central thermostat resulted in a 0.3 degrees to 0.5 degrees C reduction of the core body temperature. Fed ad libitum, Hcrt-UCP2 transgenic mice had the same caloric intake as their wild-type littermates but had increased energy efficiency and a greater median life span (12% increase in males; 20% increase in females). Thus, modest, sustained reduction of core body temperature prolonged life span independent of altered diet or CR. PMID- 17082460 TI - The polarity protein Par-3 directly interacts with p75NTR to regulate myelination. AB - Cell polarity is critical in various cellular processes ranging from cell migration to asymmetric cell division and axon and dendrite specification. Similarly, myelination by Schwann cells is polarized, but the mechanisms involved remain unclear. Here, we show that the polarity protein Par-3 localizes asymmetrically in Schwann cells at the axon-glial junction and that disruption of Par-3 localization, by overexpression and knockdown, inhibits myelination. Additionally, we show that Par-3 directly associates and recruits the p75 neurotrophin receptor to the axon-glial junction, forming a complex necessary for myelination. Together, these results point to a critical role in the establishment of cell polarity for myelination. PMID- 17082461 TI - Salary survey. U.S. life scientists report rising salaries and high job satisfaction. PMID- 17082462 TI - The role and theoretical evolution of knowledge translation and exchange in public health. AB - BACKGROUND: There is an increased emphasis in public health research on effective models and strategies to support knowledge translation (KT), the exchange, synthesis and ethically sound application of research findings within a complex set of interactions among researchers and knowledge users. In other words, KT can be seen as an acceleration of the knowledge cycle-an acceleration of the natural transformation of knowledge into use (Canadian Institutes of Health Services Research. Knowledge Translation Strategy, 2004). The most recent conceptualizations consider the complexities of public health decision-making. The role of practitioners and communities is increasingly considered. METHODS: We identify, describe and discuss the theoretical underpinnings of KT and recommend a way forward to build the evidence for more effective practice. RESULTS: Theoretical perspectives increasingly influence research on KT in public health. A range of innovative work is being conducted to explore methods for KT using practical tools, often with the support of government. CONCLUSIONS: KT describes a crucial and to date under-developed element of the research process. There is an important gap in theoretically informed empirical studies of effectiveness of proposed approaches in public health, health promotion and preventive medicine, and thus much of the debate remains abstract. There is clearly an urgent policy need to establish the effectiveness of KT models in a range of contexts. This must include both the consideration of development and the utilization of knowledge. PMID- 17082463 TI - Randomized, double-blind, placebo-controlled trial of phenylbutyrate in spinal muscular atrophy. AB - OBJECTIVE: To assess the efficacy of phenylbutyrate (PB) in patients with spinal muscular atrophy in a randomized, double-blind, placebo-controlled trial involving 10 Italian centers. METHODS: One hundred seven children were assigned to receive PB (500 mg/kg/day) or matching placebo on an intermittent regimen (7 days on/7 days off) for 13 weeks. The Hammersmith functional motor scale (primary outcome measure), myometry, and forced vital capacity were assessed at baseline and at weeks 5 and 13. RESULTS: Between January and September 2004, 107 patients aged 30 to 154 months were enrolled. PB was well tolerated, with only one child withdrawing because of adverse events. Mean improvement in functional score was 0.60 in the PB arm and 0.73 in placebo arm (p = 0.70). Changes in the secondary endpoints were also similar in the two study arms. CONCLUSIONS: Phenylbutyrate was not effective at the regimen, schedule, and duration used in this study. PMID- 17082464 TI - Projected number of people with Parkinson disease in the most populous nations, 2005 through 2030. AB - Based on published prevalence studies, we used two different methodologies to project the number of individuals with Parkinson disease (PD) in Western Europe's 5 most and the world's 10 most populous nations. The number of individuals with PD over age 50 in these countries was between 4.1 and 4.6 million in 2005 and will double to between 8.7 and 9.3 million by 2030. PMID- 17082465 TI - Graded motor imagery for pathologic pain: a randomized controlled trial. AB - BACKGROUND: Phantom limb and complex regional pain syndrome type 1 (CRPS1) are characterized by changes in cortical processing and organization, perceptual disturbances, and poor response to conventional treatments. Graded motor imagery is effective for a small subset of patients with CRPS1. OBJECTIVE: To investigate whether graded motor imagery would reduce pain and disability for a more general CRPS1 population and for people with phantom limb pain. METHODS: Fifty-one patients with phantom limb pain or CRPS1 were randomly allocated to motor imagery, consisting of 2 weeks each of limb laterality recognition, imagined movements, and mirror movements, or to physical therapy and ongoing medical care. RESULTS: There was a main statistical effect of treatment group, but not diagnostic group, on pain and function. The mean (95% CI) decrease in pain between pre- and post-treatment (100 mm visual analogue scale) was 23.4 mm (16.2 to 30.4 mm) for the motor imagery group and 10.5 mm (1.9 to 19.2 mm) for the control group. Improvement in function was similar and gains were maintained at 6 month follow-up. CONCLUSION: Motor imagery reduced pain and disability in these patients with complex regional pain syndrome type I or phantom limb pain, but the mechanism, or mechanisms, of the effect are not clear. PMID- 17082466 TI - New-onset temporal lobe epilepsy in children: lesion on MRI predicts poor seizure outcome. AB - OBJECTIVE: To determine factors predictive of long-term seizure outcome in children with new-onset temporal lobe epilepsy (TLE). METHODS: A community-based cohort of 77 children with new-onset TLE, including 14 with possible TLE, were followed prospectively with formal review 7 and 14 years following seizure onset. Diagnoses were re-evaluated at each review, and changed when new clinical, EEG, or imaging data were compelling. RESULTS: Sixty-four patients sustained the diagnosis of TLE over time; two were lost to follow-up. Age at follow-up was 12 to 29 years (median 20 years). Median follow-up was 13.7 years, 95% being followed for greater than 10 years. Nineteen patients were seizure free (SF) and off treatment, having not had seizures for 5 to 15 years. Duration of active TLE in the SF group was 1 to 8 years, the children being treated with 0 to 3 antiepileptic drugs (AEDs). Forty-three patients were not seizure free (NSF) and had ongoing seizures or had undergone epilepsy surgery. These children were treated with 1 to 10 AEDs. Fifteen NSF patients experienced 22 nonterminal seizure remissions of 1 to 7 years duration. Seventeen children had a significant antecedent to TLE. Lesions were identified on neuroimaging in 28 and included hippocampal sclerosis (HS) in 10, tumor in 8, and dysplasia in 7. All children with lesions on MRI were NSF (p < 0.001). Focal slowing on EEG was also associated with persistent seizures (p = 0.05), although this was correlated with a lesion on MRI. Infantile onset of epilepsy, family history of seizures, initial seizure frequency, antecedents, and early seizure remissions were not predictive of seizure outcome. CONCLUSION: Seizures spontaneously remit in approximately one third of children with new-onset TLE. A lesion on MRI predicts intractable seizures in TLE and the potential need for epilepsy surgery. PMID- 17082467 TI - Genotype-phenotype associations for ARX gene duplication in X-linked mental retardation. AB - We screened 165 mentally retarded patients for ARX gene 428-451 base pair (bp) duplication. Eighteen individuals from five families were found to carry the duplication, and all had intellectual impairment. Twelve presented with focal hand dystonia, while six patients had EEG abnormalities including seizures. Other symptoms included speech difficulties (4/18), testis enlargement (4/18), lower limb spasticity or foot dystonia (4/18), and facial telangiectasia (3/18). These features confirm the pleiotropic effect of the duplication. PMID- 17082468 TI - A Phase II study targeting amyloid-beta with 3APS in mild-to-moderate Alzheimer disease. AB - BACKGROUND: As a potential disease-modifying treatment for Alzheimer disease (AD), 3-amino-1-propanesulfonic acid (3APS) is a compound that binds to amyloid beta (Abeta), a toxic protein known to aggregate, leading to amyloid plaque deposition in the brain. METHODS: We assessed the safety, tolerability, and pharmacokinetic/pharmacodynamic effect of 3APS in a randomized, double-blind, placebo-controlled Phase II study in which 58 subjects with mild-to-moderate AD were randomly assigned to receive placebo or 3APS 50, 100, or 150 mg BID for 3 months. At the end of the double-blind phase, 42 of these subjects entered an open-label phase in which they received 3APS 150 mg BID for 17 months. Assessments included plasma and CSF 3APS concentrations, CSF levels of Abeta (Abeta(40) and Abeta(42)), and total tau, as well as cognitive (Alzheimer's Disease Assessment Scale-cognitive subscale, Mini-Mental State Examination) and clinical (Clinical Dementia Rating scale-Sum of Boxes) measures. RESULTS: 3APS had no significant impact on vital signs or laboratory test values. The most frequent side effects were nausea, vomiting, and diarrhea, which were intermittent and mild to moderate in severity. Seven 3APS-treated subjects discontinued because of side effects (all causalities) over the course of the study, and there were no 3APS-related serious adverse events. 3APS crossed the blood-brain barrier, and dose-dependently reduced CSF Abeta(42) levels after 3 months of treatment. There were no psychometric score differences between groups over the 3-month double-blind period. CONCLUSION: Long-term administration of 3 amino-1-propanesulfonic acid is safe, tolerated and reduces CSF Abeta(42) levels in patients with mild-to-moderate Alzheimer disease. PMID- 17082469 TI - Alpha-adducin polymorphism, atherosclerosis, and cardiovascular and cerebrovascular risk. AB - BACKGROUND AND PURPOSE: Carriers of the 460Trp allele of the alpha-adducin gene (ADD1) show higher rates of sodium reabsorption compared with homozygous carriers of the Gly460 allele and were found to have an increased risk of hypertension and cardiovascular disease. We studied the association between the Gly460Trp polymorphism and atherosclerosis, cardiovascular disease, and cerebrovascular disease. METHODS: Intima-media thickness of the common carotid artery, as well as incident stroke and myocardial infarction, were studied within 6471 subjects of the Rotterdam Study. Within 1018 subjects of the Rotterdam Scan Study, prevalent silent brain infarcts and cerebral white matter lesions were studied. Subjects were grouped into 460Trp carriers (variant carriers) and homozygous carriers of the Gly460 allele (reference). RESULTS: Intima-media thickness of the common carotid artery was 0.80 mm in variant carriers compared with 0.79 mm in the reference group (P=0.04). Variant carriers had an increased risk of any stroke (hazard ratio [HR], 1.22; 95% CI, 1.02 to 1.45), ischemic stroke (HR, 1.29; 95% CI, 1.02 to 1.63), hemorrhagic stroke (HR, 1.07; 95% CI, 0.59 to 1.92), and of myocardial infarction (HR, 1.33; 95% CI, 1.05 to 1.69). For any ischemic stroke, there was a significant interaction between the Gly460Trp polymorphism and hypertension. Variant carriers more often had a silent brain infarct (odds ratio, 1.36; 95% CI, 0.98 to 1.88) and had more subcortical white matter lesions than the reference group (1.45 vs1.24 mL; P=0.22). CONCLUSIONS: The Gly460Trp polymorphism is associated with atherosclerosis, cardiovascular disease, and cerebrovascular disease, especially in hypertensive subjects. PMID- 17082470 TI - Intra- and interrater reliability of ischemic lesion volume measurements on diffusion-weighted, mean transit time and fluid-attenuated inversion recovery MRI. AB - BACKGROUND AND PURPOSE: We investigated the intra- and interrater reliability of ischemic lesion volumes measurements assessed by different MRI sequences at various times from onset. METHODS: Ischemic lesion volumes were measured for intrarater reliability using diffusion-weighted (DWI), mean transit time (MTT) perfusion and fluid-attenuated inversion recovery (FLAIR) MRI at chronic (>3 days from stroke onset) time points. A single intrarater reader, blind to clinical information and time point, repeated the volume measurements on two occasions separated by at least 1 week. Interrater reliability was also obtained in the second set of patients using acute DWI, MTT and chronic FLAIR MRI. Four blinded readers performed these volume measurements. Average deviations across repeat measurements per lesion and differences between sample means between the two measurements were calculated globally, ie, across all sequences and time points, and per reader type for each sequence at each time point. RESULTS: There was good concordance of the mean sample volumes of the 2 intrarater readings (deviations were <4% and 2 mL globally, <2% and 2 mL for DWI, <6% and 7 mL for MTT, and <2% and 1 mL for FLAIR). There was also good concordance of the interrater readings (<5% and 2 mL globally). CONCLUSIONS: Repeat measurements of stroke lesion volumes show excellent intra- and interrater concordance for DWI, MTT and FLAIR at acute through chronic time points. PMID- 17082471 TI - Immunomodulation strategies for preventing vascular disease of the brain and heart: workshop summary. AB - This workshop examined the opportunities for translational research directed at immune and inflammatory mechanisms. This summary presents the background data in 3 general areas: (1) inflammation and hemostasis in cerebrovascular and cardiovascular disease, (2) immune interactions in the central nervous system and heart, and (3) translation of immune modulation in the brain and heart, all of which supported a consensus derivation of the opportunities for future research in these areas. The summary concludes with 11 recommendations. PMID- 17082472 TI - C-reactive protein, carotid atherosclerosis, and cerebral small-vessel disease: results of the Austrian Stroke Prevention Study. AB - BACKGROUND AND PURPOSE: C-reactive protein (CRP) is an inflammatory marker known to be a risk factor for stroke. We examined the associations between CRP, carotid atherosclerosis, white matter lesions, and lacunes as manifestations of cerebral large- and small-vessel disease. METHODS: In the community-based Austrian Stroke Prevention Study, CRP concentrations were measured by a highly sensitive assay in 700 participants at baseline. All underwent carotid duplex scanning, and a subset of 505 subjects underwent brain magnetic resonance imaging. Imaging was repeated after 3 and 6 years. We graded carotid atherosclerosis in both common and internal carotid arteries on a 5-point scale and calculated the sum of scores as an index of the severity of carotid atherosclerosis. The volume of white matter lesions and the number of lacunes were considered small vessel disease-related brain abnormalities. RESULTS: After adjustment for vascular risk factors, the severity and progression of extracranial carotid atherosclerosis increased with increasing quintiles of CRP. Only study participants in the fourth and fifth quintile (>2.50 mg/L) had significantly more baseline atherosclerosis and greater progression when we used the first quintile (<0.80 mg/L) as a reference. No interactions were seen between CRP quintiles and vascular risk factors for carotid atherosclerosis. The associations between severity and progression of small vessel disease-related brain abnormalities and CRP were nonsignificant. CONCLUSIONS: We found evidence for differential effects of CRP in different beds of the arterial brain supply. CRP was a marker for active carotid atherosclerosis but not for small vessel disease-related brain lesions. PMID- 17082473 TI - Risk of coronary and other nonstroke vascular death in relation to the presence and extent of atherosclerotic disease at the carotid bifurcation. AB - BACKGROUND AND PURPOSE: It has been proposed that purely unilateral carotid stenosis is due mainly to local hemodynamic factors, whereas more diffuse disease reflects a systemic as well as a local predisposition and should therefore be associated with higher rates of arterial disease in the other territories. However, few studies have tested this hypothesis. METHODS: We studied angiograms from 2741 patients with a recently symptomatic carotid stenosis from the European Carotid Surgery Trial and categorized them as showing purely unilateral disease (no convincing plaque in the contralateral carotid artery) or bilateral disease. We related the presence and extent of plaque at both bifurcations to the prevalence of past atherothrombotic disease in other arterial territories and the risk of future nonstroke vascular events. RESULTS: The degree of symptomatic carotid stenosis was unrelated to either the prevalence of symptomatic arterial disease in other territories at baseline or to the risk of nonstroke vascular death during follow-up, whereas there were strong associations with the presence of bilateral carotid disease. In multivariate analyses, bilateral carotid disease (n=2076) was associated with previous myocardial infarction (odds ratio [OR]=1.7; 95% CI, 1.2 to 2.4) or peripheral vascular disease (OR=1.5; 95% CI, 1.2 to 2.0) and with subsequent nonstroke vascular death (hazard ratio=2.0; 95% CI, 1.5 to 2.6). The 5-year risk of nonstroke vascular death ranged from 2.7% (95% CI, 1.5% to 4.6%) in patients with no history of coronary or peripheral arterial disease at baseline and purely unilateral carotid disease to 21.4% (95% CI, 17.6% to 26.1%) in those with bilateral carotid disease and a history of either coronary or peripheral arterial disease (P<0.0001). CONCLUSIONS: Bilateral carotid disease is associated with higher rates of symptomatic arterial disease in other territories, suggesting a systemic predisposition to atherosclerosis and hence, the need for more aggressive preventive treatment, whereas purely unilateral carotid disease indicates a mainly local hemodynamic etiology. PMID- 17082474 TI - Granulocyte-colony-stimulating factor mobilizes bone marrow stem cells in patients with subacute ischemic stroke: the Stem cell Trial of recovery EnhanceMent after Stroke (STEMS) pilot randomized, controlled trial (ISRCTN 16784092). AB - BACKGROUND AND PURPOSE: Loss of motor function is common after stroke and leads to significant chronic disability. Stem cells are capable of self-renewal and of differentiating into multiple cell types, including neurones, glia, and vascular cells. We assessed the safety of granulocyte-colony-stimulating factor (G-CSF) after stroke and its effect on circulating CD34+ stem cells. METHODS: We performed a 2-center, dose-escalation, double-blind, randomized, placebo controlled pilot trial (ISRCTN 16784092) of G-CSF (6 blocks of 1 to 10 microg/kg SC, 1 or 5 daily doses) in 36 patients with recent ischemic stroke. Circulating CD34+ stem cells were measured by flow cytometry; blood counts and measures of safety and functional outcome were also monitored. All measures were made blinded to treatment. RESULTS: Thirty-six patients, whose mean+/-SD age was 76+/-8 years and of whom 50% were male, were recruited. G-CSF (5 days of 10 microg/kg) increased CD34+ count in a dose-dependent manner, from 2.5 to 37.7 at day 5 (area under curve, P=0.005). A dose-dependent rise in white cell count (P<0.001) was also seen. There was no difference between treatment groups in the number of patients with serious adverse events: G-CSF, 7/24 (29%) versus placebo 3/12 (25%), or in their dependence (modified Rankin Scale, median 4, interquartile range, 3 to 5) at 90 days. CONCLUSIONS: G-CSF is effective at mobilizing bone marrow CD34+ stem cells in patients with recent ischemic stroke. Administration is feasible and appears to be safe and well tolerated. The fate of mobilized cells and their effect on functional outcome remain to be determined. PMID- 17082475 TI - Is sex a prognostic factor in stroke rehabilitation? A matched comparison. AB - BACKGROUND AND PURPOSE: We sought to assess the specific influence of sex on rehabilitation results. METHODS: A case-control study in 440 consecutive patients with sequelae of first ischemic stroke were enrolled in 2 subgroups (males and females) and matched for severity of stroke (evaluated by means of the Canadian Neurological Scale), age (within 1 year), and onset-admission interval (within 3 days). Functional data, evaluated by means of the Barthel Index and the Rivermead Mobility Index, were compared between subgroups. Logistic regressions were used to clarify the role of sex in affecting global autonomy and mobility. RESULTS: After rehabilitation treatment, a sex-related difference was observed essentially in the higher levels of response. Indeed, more men than women reached independence in both stair climbing and activities of daily living (ADL), with a higher response and effectiveness on mobility. In multivariate analyses, male patients had a 3 times higher probability than female patients of good autonomy in both stair climbing and ADL (odds ratio [OR]=3.22; 95% CI, 1.67 to 6.18 and OR=2.92; 95% CI, 1.63 to 5.42, respectively). Conversely, female patients had a higher risk of walking with a cane (OR=1.69; 95%, CI 1.04 to 2.76) or of partial autonomy with respect to ADL (OR=1.90; 95% CI, 1.25 to 2.91). No significant difference was found for the other functional parameters. CONCLUSIONS: Female sex is a mildly unfavorable prognostic factor in rehabilitation results after stroke. PMID- 17082476 TI - Transcriptional silencing of the death gene BNIP3 by cooperative action of NF kappaB and histone deacetylase 1 in ventricular myocytes. AB - Earlier we identified a survival role for NF-kappaB in ventricular myocytes, however, the underlying mechanism was undefined. In this report we provide new mechanistic evidence that the hypoxia-inducible death factor BNIP3 is transcriptionally silenced by NF-kappaB through a mechanism that involves the cooperative actions of HDAC1. Activation of the NF-kappaB signaling pathway in ventricular myocytes suppressed basal and hypoxia-inducible BNIP3 gene activity. Basal Bnip3 gene expression was increased in cells derived from p65(-/-) deficient mice. The histone deacetylase (HDAC) inhibitor Trichostatin A (TSA 10 nM) suppressed the inhibitory actions of NF-kappaB on Bnip3 gene transcription. Basal and hypoxia- induced Bnip3 transcription was repressed by wild type but not a catalytically inactive mutant of HDAC1. Immunoprecipitation assays verified interaction of HDAC1 with wild type p65 NF-kappaB and mutations of p65 defective for transactivation in ventricular myocytes. Deletion analysis revealed canonical NF-kappaB elements within the Bnip3 promoter to be important for repression of Bnip3 gene expression by HDAC1. Further, the ability of HDAC1 to repress Bnip3 gene transcription was lost in cells derived from p65(-/-) deficient mice but was restored by repletion of p65 NF-kappaB into p65(-/-) cells. Mutations of p65 NF kappaB defective for DNA binding but not for transactivation abrogated the inhibitory actions of HDAC1 on the Bnip3 gene transcription. Together, our findings provide new mechanistic insight into the cytoprotective actions conferred by NF-kappaB that extend to the active transcriptional repression of the death factor Bnip3 through a mechanism that is mutually dependent on HDAC-1. PMID- 17082477 TI - Haptoglobin genotype is a regulator of reverse cholesterol transport in diabetes in vitro and in vivo. AB - Two common alleles exist at the haptoglobin (Hp) locus, and the Hp2 allele is associated with an increased incidence of cardiovascular disease, specifically in diabetes mellitus (DM). Oxidative stress is increased in Hp2 mice and humans with DM. Oxidative modification of the apolipoprotein A-I inhibits reverse cholesterol transport. We sought to test the hypothesis that reverse cholesterol transport is impaired in Hp2 DM mice and humans. In vitro, using serum from non-DM and DM individuals, we measured cholesterol efflux from (3)H-cholesterol-labeled macrophages. In vivo, we injected (3)H-cholesterol-loaded macrophages intraperitoneally into non-DM and DM mice with the Hp1-1 or Hp2-2 genotype and monitored (3)H-tracer levels in plasma, liver, and feces. In vitro, in DM individuals only, we observed significantly decreased cholesterol efflux from macrophages incubated with serum from Hp2-1 or Hp2-2 as compared with Hp1-1 individuals (P<0.01). The interaction between Hp type and DM was recapitulated using purified Hp and glycated Hb. In vivo, DM mice loaded with (3)H-cholesterol labeled macrophages had a 40% reduction in (3)H-cholesterol in plasma, liver, and feces as compared with non-DM mice (P<0.01). The reduction in reverse cholesterol transport associated with DM was significantly greater in Hp2-2 mice as compared with Hp1-1 mice (54% versus 25% in plasma; 52% versus 27% in liver; 57% versus 32% in feces; P<0.03). reverse cholesterol transport is decreased in Hp2-2 DM. This may explain in part the increased atherosclerotic burden found in Hp2-2 DM individuals. PMID- 17082478 TI - Differential expression of ion channel transcripts in atrial muscle and sinoatrial node in rabbit. AB - The aim of the study was to identify ion channel transcripts expressed in the sinoatrial node (SAN), the pacemaker of the heart. Functionally, the SAN can be divided into central and peripheral regions (center is adapted for pacemaking only, whereas periphery is adapted to protect center and drive atrial muscle as well as pacemaking) and the aim was to study expression in both regions. In rabbit tissue, the abundance of 30 transcripts (including transcripts for connexin, Na(+), Ca(2+), hyperpolarization-activated cation and K(+) channels, and related Ca(2+) handling proteins) was measured using quantitative PCR and the distribution of selected transcripts was visualized using in situ hybridization. Quantification of individual transcripts (quantitative PCR) showed that there are significant differences in the abundance of 63% of the transcripts studied between the SAN and atrial muscle, and cluster analysis showed that the transcript profile of the SAN is significantly different from that of atrial muscle. There are apparent isoform switches on moving from atrial muscle to the SAN center: RYR2 to RYR3, Na(v)1.5 to Na(v)1.1, Ca(v)1.2 to Ca(v)1.3 and K(v)1.4 to K(v)4.2. The transcript profile of the SAN periphery is intermediate between that of the SAN center and atrial muscle. For example, Na(v)1.5 messenger RNA is expressed in the SAN periphery (as it is in atrial muscle), but not in the SAN center, and this is probably related to the need of the SAN periphery to drive the surrounding atrial muscle. PMID- 17082479 TI - A novel signal transduction cascade involving direct physical interaction of the renin/prorenin receptor with the transcription factor promyelocytic zinc finger protein. AB - A human renin/prorenin receptor (RER) has recently been cloned. To gain insight into the molecular function of the RER, we studied its signal transduction mechanisms. Initially, we found a ubiquitous and intracellular expression pattern of the human RER. Consistently, we observed several transcriptional start sites and a high promoter activity of the human RER. We could identify the transcription factor promyelocytic zinc finger (PLZF) protein as a direct protein interaction partner of the C-terminal domain of the RER by yeast 2-hybrid screening and coimmunoprecipitation. Coimmunoprecipitation experiments also indicated homodimerization of the RER. On activation of the RER by renin, PLZF is translocated into the nucleus and represses transcription of the RER itself, thereby creating a very short negative feedback loop, but activates transcription of the p85alpha subunit of the phosphatidylinositol-3 kinase (PI3K-p85alpha). Small interfering RNA against the RER abolished these effects. A PLZF cis-element in the RER promoter was identified by site-directed mutagenesis and electrophoretic mobility-shift assay. Renin stimulation caused a 6-fold recruitment of PLZF to this promoter region as shown by chromatin immunoprecipitation. Moreover, renin stimulation of rat H9c2 cardiomyoblasts induced an increase of cell number and a decrease of apoptosis. These effects were partly abolished by PI3K inhibition and completely abrogated by small interfering RNA against PLZF. Finally, experiments in PLZF knockout mice confirmed the role of PLZF as an upstream regulator of RER and PI3K-p85alpha. Our data demonstrate the existence of a novel signal transduction pathway involving the ligand renin, RER, and the transcription factor PLZF, which is of physiological and putative pathophysiological relevance. PMID- 17082480 TI - Altered Na+ channels promote pause-induced spontaneous diastolic activity in long QT syndrome type 3 myocytes. AB - Long QT syndrome (LQTS) type 3 (LQT3), typified by the DeltaKPQ mutation (LQT3 mutation in which amino acid residues 1505 to 1507 [KPQ] are deleted), is caused by increased sodium entry during the action potential plateau resulting from mutation-altered inactivation of the Na(v)1.5 channel. Although rare, LQT3 is the most lethal of common LQTS variants. Here we tested the hypothesis that cellular electrical dysfunction, caused not only by action potential prolongation but also by mutation-altered Na(+) entry, distinguishes LQT3 from other LQTS variants and may contribute to its distinct lethality. We compared cellular electrical activity in myocytes isolated from mice heterozygous for the DeltaKPQ mutation (DeltaKPQ) and myocytes from wild-type littermates. Current-clamp pause protocols induced rate-dependent spontaneous diastolic activity (delayed after depolarizations) in 6 of 7 DeltaKPQ, but no wild-type, myocytes (n=11) tested. Voltage-clamp pause protocols that independently control depolarization duration and interpulse interval identified a distinct contribution of both depolarization duration and mutant Na(+) channel activity to the generation of Ca(i)(2+) dependent diastolic transient inward current. This was found at rates and depolarization durations relevant both to the mouse model and to LQT3 patients. Flecainide, which preferentially inhibits mutation-altered late Na(+) current and is used to treat LQT3 patients, suppresses transient inward current formation in voltage-clamped DeltaKPQ myocytes. Our results demonstrate a marked contribution of mutation-altered Na(+) entry to the incidence of pause-dependent spontaneous diastolic activity in DeltaKPQ myocytes and suggest that altered Na(+) entry may contribute to the elevated lethality of LQT3 versus other LQTS variants. PMID- 17082481 TI - Targeting of metallothionein by L-homocysteine: a novel mechanism for disruption of zinc and redox homeostasis. AB - OBJECTIVE: L-homocysteine and/or L-homocystine interact in vivo with albumin and other extracellular proteins by forming mixed-disulfide conjugates. Because of its extremely rich cysteine content, we hypothesized that metallothionein, a ubiquitous intracellular zinc-chaperone and superoxide anion radical scavenger, reacts with L-homocysteine and that homocysteinylated-metallothionein suffers loss of function. METHODS AND RESULTS: 35S-homocysteinylated-metallothionein was resolved in lysates of cultured human aortic endothelial cells in the absence and presence of reduced glutathione by SDS-PAGE and identified by Western blotting and phosphorimaging. Using zinc-Sepharose chromatography, L-homocysteine was shown to impair the zinc-binding capacity of metallothionein even in the presence of reduced glutathione. L-Homocysteine induced a dose-dependent increase in intracellular free zinc in zinquin-loaded human aortic endothelial cells within 30 minutes, followed by the appearance of early growth response protein-1 within 60 minutes. In addition, intracellular reactive oxygen species dramatically increased 6 hours after L-homocysteine treatment. In vitro studies demonstrated that L-homocysteine is a potent inhibitor of the superoxide anion radical scavenging ability of metallothionein. CONCLUSIONS: These studies provide the first evidence that L-homocysteine targets intracellular metallothionein by forming a mixed-disulfide conjugate and that loss of function occurs after homocysteinylation. The data support a novel mechanism for disruption of zinc and redox homeostasis. PMID- 17082482 TI - The CX3C chemokine fractalkine induces vascular dysfunction by generation of superoxide anions. AB - OBJECTIVE: The chemokine fractalkine activates platelets and induces leukocyte adhesion to the endothelium. Expression of fractalkine and its receptor, CX3CR1, is elevated in coronary artery disease. We assessed the effects of fractalkine on vascular function in isolated rat aorta. METHODS AND RESULTS: CX3CR1 expression was demonstrated in rat aortic endothelial and smooth muscle cells by immunohistochemistry, Western blot, and polymerase chain reaction (PCR). Fractalkine (up to 1 microg/mL) did not directly induce contractile or relaxant responses when applied to rat aortic rings in organ baths. Short-term incubation with fractalkine (1 microg/mL) for 5 minutes did not affect vascular reactivity. Pretreatment of isolated rat aortic rings with fractalkine for 2 hours impaired acetylcholine-induced nitric oxide (NO)-mediated relaxation after preconstriction with phenylephrine in a concentration-dependent manner. The concentration response to the NO donor DEA-NONOate was significantly shifted to the right. The radical scavenger tiron normalized the attenuated acetylcholine-induced relaxation after fractalkine incubation. Aortic superoxide formation was enhanced by fractalkine, which was inhibited by diphenyleneiodonium but not by inhibitors of xanthine oxidase or NO synthase. CONCLUSIONS: In addition to its role as a chemokine and adhesion molecule, fractalkine induces vascular dysfunction by stimulating vascular reactive oxygen species resulting in reduced NO bioavailability. PMID- 17082483 TI - Statin treatment upregulates vascular neuronal nitric oxide synthase through Akt/NF-kappaB pathway. AB - OBJECTIVE: Three-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) are known to enhance vascular expression of endothelial (eNOS) and inducible nitric oxide synthase (iNOS). In this study, we examined whether statins also upregulate vascular expression of neuronal NOS (nNOS). METHODS AND RESULTS: In cultured rat aortic smooth muscle cells, treatment with atorvastatin significantly increased nNOS expression, associated with activation of Akt and NF-kappaB. Inhibition of Akt by dominant-negative Akt suppressed atorvastatin-induced nNOS expression as well as Akt and NF-kappaB activation. Inhibition of NF-kappaB by dominant-negative IkappaB also attenuated atorvastatin induced nNOS expression and NF-kappaB activation, but not Akt activation. We further examined whether atorvastatin also enhances nNOS expression in isolated mouse aorta, and if so, how much nNOS-derived NO accounts for atorvastatin induced NOx production. In isolated aortas of wild-type mice, atorvastatin significantly increased all three NOS isoform expression and NOx production. In isolated aortas of doubly i/eNOS(-/-), n/eNOS(-/-), and n/iNOS(-/-) mice, which express only nNOS, iNOS, and eNOS, respectively, atorvastatin-induced NOx production was approximately 25%, 25%, and 50% to that of wild-type mice, respectively, suggesting that nNOS accounts for 25% of the atorvastatin-mediated NOx production. CONCLUSIONS: These results indicate that atorvastatin upregulates vascular nNOS through Akt/NF-kappaB pathway, demonstrating a novel nNOS-mediated vascular effect of the statin. PMID- 17082484 TI - Role of the Toll-like receptor 4/NF-kappaB pathway in saturated fatty acid induced inflammatory changes in the interaction between adipocytes and macrophages. AB - OBJECTIVE: Previous studies demonstrated that obese adipose tissue is characterized by increased infiltration of macrophages, suggesting that they might represent an important source of inflammation. Using an in vitro coculture system composed of 3T3-L1 adipocytes and RAW264 macrophages, we previously demonstrated that saturated fatty acids (FAs) and tumor necrosis factor (TNF) alpha derived from adipocytes and macrophages, respectively, play a major role in the coculture-induced inflammatory changes. METHODS AND RESULTS: Coculture of adipocytes and macrophages resulted in the activation of nuclear factor-kappaB (NF-kappaB), a primary regulator of inflammatory responses, in both cell types. Pharmacological inhibition of NF-kappaB markedly suppressed the coculture-induced production of proinflammatory cytokines and adipocyte lipolysis. Peritoneal macrophages obtained from Toll-like receptor 4 (TLR4) mutant mice exhibited marked attenuation of TNFalpha production in response to saturated FAs. Notably, coculture of hypertrophied adipocytes and TLR4-mutant macrophages resulted in marked inhibition of proinflammatory cytokine production and adipocyte lipolysis. We also observed that endogenous FAs, which are released from adipocytes via the beta3-adrenergic stimulation, resulted in the activation of the TLR4/NF-kappaB pathway. CONCLUSIONS: These findings suggest that saturated FAs, which are released in large quantities from hypertrophied adipocytes via the macrophage induced adipocyte lipolysis, serve as a naturally occurring ligand for TLR4, thereby inducing the inflammatory changes in both adipocytes and macrophages through NF-kappaB activation. PMID- 17082485 TI - Prothrombotic effects of hyperhomocysteinemia and hypercholesterolemia in ApoE deficient mice. AB - OBJECTIVE: We tested the hypothesis that hyperhomocysteinemia and hypercholesterolemia promote arterial thrombosis in mice. METHODS AND RESULTS: Male apolipoprotein E (Apoe)-deficient mice were fed one of four diets: control, hyperhomocysteinemic (HH), high fat (HF), or high fat/hyperhomocysteinemic (HF/HH). Total cholesterol was elevated 2-fold with the HF or HF/HH diets compared with the control or HH diets (P<0.001). Plasma total homocysteine (tHcy) was elevated (12 to 15 micromol/L) with the HH or HF/HH diets compared with the control or HF diets (4 to 6 micromol/L; P<0.001). Aortic sinus lesion area correlated strongly with total cholesterol (P<0.001) but was independent of tHcy. At 12 weeks of age, the time to thrombotic occlusion of the carotid artery after photochemical injury was >50% shorter in mice fed the HF diets, with or without hyperhomocysteinemia, compared with the control diet (P<0.05). At 24 weeks of age, carotid artery thrombosis was also accelerated in mice fed the HH diet (P<0.05). Endothelium-dependent nitric oxide-mediated relaxation of carotid artery rings was impaired in mice fed the HF, HH, or HF/HH diets compared with the control diet (P<0.05). CONCLUSIONS: Hyperhomocysteinemia and hypercholesterolemia, alone or in combination, produce endothelial dysfunction and increased susceptibility to thrombosis in Apoe-deficient mice. PMID- 17082486 TI - A central role for nicotinic cholinergic regulation of growth factor-induced endothelial cell migration. AB - OBJECTIVE: An endothelial nicotinic acetylcholine receptor (nAChR) participates in atherogenesis and tumorigenesis by promoting neovascularization. To date, the mechanisms of nAChR-mediated angiogenesis and their relationship to angiogenic factors, eg, VEGF and bFGF, are unknown. METHODS AND RESULTS: Nicotine induced dose-dependent human microvascular endothelial cell (HMVEC) migration, a key angiogenesis event, to an extent which was equivalent in magnitude to bFGF (10 ng/mL) but less than for VEGF (10 ng/mL). Unexpectedly, nAChR antagonism not only abolished nicotine-induced HMVEC migration but also abolished migration induced by bFGF and attenuated migration induced by VEGF. Transcriptional profiling identified gene expression programs which were concordantly regulated by all 3 angiogens (nicotine, VEGF, and bFGF), a notable feature of which includes corepression of thioredoxin-interacting protein (TXNIP), endogenous inhibitor of the redox regulator thioredoxin. Furthermore, TXNIP repression by all 3 angiogens induced thioredoxin activity. Silencing thioredoxin by small interference RNA abrogated all angiogen-induced migration while silencing TXNIP strongly induced HMVEC migration. Interestingly, nAChR antagonism abrogates growth factor (VEGF and bFGF)-mediated induction of thioredoxin activity. CONCLUSIONS: Nicotine promotes angiogenesis via stimulation of nAChR-dependent endothelial cell migration. Furthermore, growth factor-induced HMVEC migration, a key angiogenesis event, requires nAChR activation--an effect mediated in part by nAChR-dependent regulation of thioredoxin activity. PMID- 17082487 TI - Therapeutic angiogenesis using novel vascular endothelial growth factor-E/human placental growth factor chimera genes. AB - BACKGROUND: Vascular endothelial growth factor-A (VEGF-A) promotes angiogenesis but causes adverse side effects such as edema or tissue inflammation. VEGF-E, found in the genome of the Orf virus, specifically binds to VEGF receptor-2 and shows mitotic activity on endothelial cells. Recently, we created two forms of VEGF-E and human placental growth factor (PlGF) chimera genes (VEGF-E chimera #9 and VEGF-E chimera #33), which are humanized genes with VEGF-E function but showing less antigenicity. METHODS AND RESULTS: We examined potential proangiogenic activities of these chimera genes. Four types of expression plasmids (pCDNA3.1-LacZ, phVEGF-A, pVEGF-Echimera#9, and pVEGF-Echimera#33) were administered in a rat model of hindlimb ischemia. Either pVEGF-Echimera#9, pVEGF Echimera#33, or phVEGF-A significantly increased the ratio of ischemic/normal hindlimb blood-flow compared with the control pCDNA3.1-LacZ treated group (by 1.5 fold, 1.5-fold, and 1.4-fold, respectively, P<0.05). Histochemical staining by alkaline phosphatase also revealed that either pVEGF-Echimera#9, pVEGF Echimera#33, or phVEGF-A increased the capillary density compared with the pCDNA3.1-LacZ treated group (1.4-fold, 1.5-fold, and 1.5-fold, respectively, P<0.05). Furthermore, immunostaining for anti-ED1 revealed that fewer macrophages had infiltrated in both pVEGF-Echimera#9 and pVEGF-Echimera#33 groups compared with the phVEGF-A group (P<0.05). CONCLUSIONS: Novel VEGF-E/human PlGF chimera genes, pVEGF-Echimera#9, and pVEGF-Echimera#33 significantly stimulated angiogenesis in response to tissue ischemia to an almost identical extent to that induced by phVEGF-A with fewer tissue inflammation responses. PMID- 17082488 TI - Biomarkers of atherosclerotic plaque instability and rupture. AB - Basic research over the last two decades has identified a large number of molecules pertinent to the atherosclerotic process, which have clearly improved our understanding of the underlying pathology. It is now well established that inflammation represents a major feature which is present in the vessel wall throughout all stages of the disease until the final pathophysiologic steps, representing plaque destabilization and eventually plaque rupture. Several cells typical for the atherosclerotic plaque, like monocyte-derived macrophages and T lymphocytes are able to produce and secrete such mediator molecules, like cytokines, chemokines, growth-factors, enzymes, and disintegrins, which lead to activation of endothelial cells, proliferation of smooth muscle cells, lesion progression, and finally to the weakening of a vulnerable plaque by matrix degradation of its fibrous cap. Today, many of these molecules involved can be measured systemically by sensitive assays, and elevated concentrations in the circulation have been shown to be associated with future cardiovascular events. Determination of several of these molecules carries important prognostic information, independent of traditional risk factors, and may turn out to be useful in improving risk stratification. However, for most of these biomarkers the clinical utility has not yet been established. PMID- 17082489 TI - Low circulating endogenous secretory receptor for AGEs predicts cardiovascular mortality in patients with end-stage renal disease. AB - OBJECTIVE: Receptor for advanced glycation end-products (RAGE) is involved in diabetic vascular complications. We have recently shown that plasma endogenously secretory RAGE (esRAGE), an alternatively spliced form of RAGE, is closely associated with metabolic syndrome and atherosclerosis. Here, we evaluated if plasma esRAGE is a predictor of cardiovascular mortality in a cohort of 206 (171 nondiabetic) patients with end-stage renal diseases (ESRD). METHODS AND RESULTS: The cohort was followed for a median of 111 months, and 74 deaths including 34 cardiovascular deaths were recorded. Plasma esRAGE was measured at baseline. Cumulative incidence of cardiovascular death by Kaplan-Meier estimation was significantly higher in subjects in the lowest tertile of plasma esRAGE than those in the middle or the highest tertile both in all and nondiabetic subjects alone. In all subjects, as compared with the lowest tertile of plasma esRAGE, the hazards ratios for the highest and middle tertile were 0.40 (95% CI, 0.18 to 0.89) and 0.26 (0.10 to 0.66), respectively. The higher risk for lower esRAGE was still significant even after adjusted either with body mass index, hypertension, dyslipidemia and vascular complications, but was confounded by age and diabetes. CONCLUSIONS: Low circulating esRAGE is a predictor for cardiovascular mortality in ESRD patients. PMID- 17082490 TI - Increased plasma oxidized phospholipid:apolipoprotein B-100 ratio with concomitant depletion of oxidized phospholipids from atherosclerotic lesions after dietary lipid-lowering: a potential biomarker of early atherosclerosis regression. AB - BACKGROUND: Oxidized phospholipids (OxPL) are pro-inflammatory. We evaluated whether changes in plasma levels of OxPL associated with apolipoprotein B-100 (apoB-100) reflect changes in OxPL content in atherosclerotic plaques during dietary-induced atherosclerosis progression and regression. METHODS AND RESULTS: OxPL content was measured in plasma and immunohistochemically in aortic plaques with antibody E06 in cynomolgus monkeys and New Zealand White rabbits at baseline, after a high-fat/high-cholesterol diet and after reversion to normal chow. The OxPL/apoB ratio, representing the content of OxPL on individual apoB 100 particles, and Total apoB-OxPL (OxPL/apoB multiplied by plasma apoB levels), reflecting the OxPL content on all apoB-100 particles, were measured. Total apoB OxPL plasma levels increased 3-fold (P<0.0001) during hypercholesterolemia and decreased approximately 75% (P<0.0001) during reversion to normocholesterolemia. In contrast, OxPL/apoB levels decreased significantly (P<0.0001) during hypercholesterolemia and increased significantly (P=0.0002) during reversion to normocholesterolemia. Immunostaining revealed that during atherosclerosis progression OxPL co-localized with apoB-100, whereas during regression OxPL virtually disappeared. CONCLUSIONS: In the setting of overall reduction of plasma OxPL levels after dietary lipid-lowering, increases in the OxPL/apoB ratio reflect reduced content of OxPL in atherosclerotic plaques. These data suggest that changes in the OxPL/apoB ratio may reflect early atherosclerosis regression. PMID- 17082491 TI - Nox4 is required for maintenance of the differentiated vascular smooth muscle cell phenotype. AB - OBJECTIVE: The mechanisms responsible for maintaining the differentiated phenotype of adult vascular smooth muscle cells (VSMCs) are incompletely understood. Reactive oxygen species (ROS) have been implicated in VSMC differentiation, but the responsible sources are unknown. In this study, we investigated the role of Nox1 and Nox4-derived ROS in this process. METHODS AND RESULTS: Primary VSMCs were used to study the relationship between Nox homologues and differentiation markers such as smooth muscle alpha-actin (SM alpha-actin), smooth muscle myosin heavy chain (SM-MHC), heavy caldesmon, and calponin. We found that Nox4 and differentiation marker genes were downregulated from passage 1 to passage 6 to 12, whereas Nox1 was gradually upregulated. Nox4 co-localized with SM alpha-actin-based stress fibers in differentiated VSMC, and moved into focal adhesions in de-differentiated cells. siRNA against nox4 reduced NADPH driven superoxide production in serum-deprived VSMCs and downregulated SM-alpha actin, SM-MHC, and calponin, as well as SM-alpha actin stress fibers. Nox1 depletion did not decrease these parameters. CONCLUSIONS: Nox4-derived ROS are critical to the maintenance of the differentiated phenotype of VSMCs. These findings highlight the importance of identifying the specific source of ROS involved in particular cellular functions when designing therapeutic interventions. PMID- 17082492 TI - Latent tuberculosis infection treatment and T-cell responses to Mycobacterium tuberculosis-specific antigens. AB - RATIONALE: There is currently no available test for monitoring the effect of treatment of latent tuberculosis infection (LTBI) to indicate cure or predict risk of subsequent progression to disease. OBJECTIVE: We used the T-SPOT.TB assay, which measures T-cell interferon-gamma responses to the Mycobacterium tuberculosis-specific peptides early secretory antigenic target 6-kD protein (ESAT-6) and culture filtrate protein 10 (CFP-10), to determine the effect of LTBI treatment on these responses. METHODS: A total of 226 tuberculosis contacts with positive T-SPOT.TB results underwent repeat testing on LTBI treatment completion. The majority (96%) received 6 months of isoniazid. The pre- and post treatment T-SPOT.TB results were analyzed according to the combined and separate responses to ESAT-6 and CFP-10 antigens. RESULTS: The T-SPOT.TB reverted to negative in 85 (37.6%) contacts at treatment completion. Treatment had a significant effect on the response to CFP-10 (p < 0.001; reversion rate, 48.6%), but not on the response to ESAT-6 (p = 0.081; reversion rate, 21.6%). The median number of spot-forming cells (SFCs)/2.5 x 10(5) peripheral blood mononuclear cells (PBMCs) pre- and post-treatment was 6 versus 4.5 for ESAT-6 (p = 0.116) and 11 versus 4 for CFP-10 (p < 0.001). There was a significant difference between the change (fall) in the pre- and post-treatment responses to CFP-10 (6 SFCs/2.5 x 10(5) PBMCs) and ESAT-6 (0 SFCs/2.5 x 10(5) PBMCs; p < 0.001). Significantly different age-related T-cell responses to the two antigens were found. CONCLUSION: LTBI treatment had a differential effect on T-cell responses to ESAT 6 and CFP-10 as measured by the T-SPOT.TB. The quantitative response to CFP-10 may be a useful LTBI treatment-monitoring tool. PMID- 17082493 TI - Coagulation factor Xa modulates airway remodeling in a murine model of asthma. AB - RATIONALE: Previous studies have demonstrated that dysregulated coagulation and fibrinolysis contribute to the pathogenesis of asthma. OBJECTIVE: The role of procoagulant factor X in a murine model of ovalbumin (OVA)-induced asthma was investigated. METHODS: Biochemical, cellular, and physiologic in vivo and in vitro approaches were used to determine effects of factor X on the asthmatic response in mice. MEASUREMENTS AND MAIN RESULTS: Factor X transcript levels and factor Xa activity were increased in lungs of asthmatic mice challenged with OVA, compared with controls treated with phosphate-buffered saline. Factor X was highly expressed in bronchoalveolar lavage fluid macrophages from asthmatic mice. Treatment of mice with the factor Xa inhibitor fondaparinux during the last 4 wk of OVA challenge resulted in the attenuation of airway hyperresponsiveness but did not alter infiltration of inflammatory cells into the lung. There was a significant decrease in the thickness of the mucosal layer and in lung collagen deposition in fondaparinux-treated mice. In vitro investigations using human mucus-producing NCI-H292 cells indicated that exogenous factor Xa enhanced mucin production in a dose-dependent manner. Levels of amphiregulin, a protein that induces mucin production, were also increased in cells stimulated by factor Xa. CONCLUSIONS: The results of this study introduce a novel participant in the asthmatic response and indicate that factor Xa functions in airway remodeling in asthma by stimulating mucin production, through regulation of amphiregulin expression and collagen deposition. PMID- 17082494 TI - A novel host defense system of airways is defective in cystic fibrosis. AB - RATIONALE: The respiratory tract is constantly exposed to airborne microorganisms. Nevertheless, normal airways remain sterile without recruiting phagocytes. This innate immune activity has been attributed to mucociliary clearance and antimicrobial polypeptides of airway surface liquid. Defective airway immunity characterizes cystic fibrosis (CF), a disease caused by mutations in the CF transmembrane conductance regulator, a chloride channel. The pathophysiology of defective immunity in CF remains to be elucidated. OBJECTIVE: We investigated the ability of non-CF and CF airway epithelia to kill bacteria through the generation of reactive oxygen species (ROS). METHODS: ROS production and ROS-mediated bactericidal activity were determined on the apical surfaces of human and rat airway epithelia and on cow tracheal explants. MEASUREMENTS AND MAIN RESULTS: Dual oxidase enzyme of airway epithelial cells generated sufficient H(2)O(2) to support production of bactericidal hypothiocyanite (OSCN(-)) in the presence of airway surface liquid components lactoperoxidase and thiocyanate (SCN(-)). This OSCN(-) formation eliminated Staphylococcus aureus and Pseudomonas aeruginosa on airway mucosal surfaces, whereas it was nontoxic to the host. In contrast to normal epithelia, CF epithelia failed to secrete SCN(-), thereby rendering the oxidative antimicrobial system inactive. CONCLUSIONS: These data indicate a novel innate defense mechanism of airways that kills bacteria via ROS and suggest a new cellular and molecular basis for defective airway immunity in CF. PMID- 17082495 TI - Higher urine nitric oxide is associated with improved outcomes in patients with acute lung injury. AB - RATIONALE: Nitrogen oxide (NO) species are markers for oxidative stress that may be pathogenic in acute lung injury (ALI). OBJECTIVES: We tested two hypotheses in patients with ALI: (1) higher levels of urine NO would be associated with worse clinical outcomes, and (2) ventilation with lower VT would reduce urine NO as a result of less stretch injury. METHODS: Urine NO levels were measured by chemiluminescence in 566 patients enrolled in the National Heart Lung and Blood Institute Acute Respiratory Distress Syndrome Network trial of 6 ml/kg versus 12 ml/kg VT ventilation. The data were expressed corrected and uncorrected for urine creatinine (Cr). RESULTS: Higher baseline levels of urine NO to Cr were associated with lower mortality (odds ratio, 0.43 per log(10) increase in the ratio), more ventilator-free days (mean increase, 1.9 d), and more organ-failure free days (mean increase, 2.3 d) on multivariate analysis (p < 0.05 for all analyses). Similar results were obtained using urine NO alone. NO to Cr levels were higher on Day 3 in the 6 ml/kg than in the 12 ml/kg VT group (p = 0.04). CONCLUSIONS: Contrary to our hypothesis, higher urine NO was associated with improved outcomes in ALI at baseline and after treatment with the 6 ml/kg VT strategy. Higher endogenous NO may reflect less severe lung injury and better preservation of the pulmonary and systemic endothelium or may serve a protective function in patients with ALI. PMID- 17082496 TI - Caspase-3 regulation of diaphragm myonuclear domain during mechanical ventilation induced atrophy. AB - RATIONALE: Unloading the diaphragm via mechanical ventilation (MV) results in rapid diaphragmatic fiber atrophy. It is unknown whether the myonuclear domain (cytoplasmic myofiber volume/myonucleus) of diaphragm myofibers is altered during MV. OBJECTIVE: We tested the hypothesis that MV-induced diaphragmatic atrophy is associated with a loss of myonuclei via a caspase-3-mediated, apoptotic-like mechanism resulting in a constant myonuclear domain. METHODS: To test this postulate, Sprague-Dawley rats were randomly assigned to a control group or to experimental groups exposed to 6 or 12 h of MV with or without administration of a caspase-3 inhibitor. MEASUREMENTS AND MAIN RESULTS: After 12 h of MV, type I and type IIa diaphragm myofiber areas were decreased by 17 and 23%, respectively, and caspase-3 inhibition attenuated this decrease. Diaphragmatic myonuclear content decreased after 12 h of MV and resulted in the maintenance of a constant myonuclear domain in all fiber types. Both 6 and 12 h of MV resulted in caspase-3 dependent increases in apoptotic markers in the diaphragm (e.g., number of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling positive nuclei and DNA fragmentation). Caspase-3-dependent increases in apoptotic markers occurred after 6 h of MV, before the onset of myofiber atrophy. CONCLUSIONS: Collectively, these data support the hypothesis that the myonuclear domain of diaphragm myofibers is maintained during prolonged MV and that caspase-3-mediated myonuclear apoptosis contributes to this process. PMID- 17082497 TI - Logic regression for analysis of the association between genetic variation in the renin-angiotensin system and myocardial infarction or stroke. AB - Recent developments in genetic sequencing technology now make it possible to genotype large numbers of single nucleotide polymorphisms (SNPs) in large samples. Many association studies using SNP data are now being carried out. Typically, these observational studies establish whether certain haplotypes or individual SNPs are associated with a health outcome. Few methods exist for finding interaction effects among multiple SNPs or between SNPs and environmental factors. In this paper, the authors describe logic regression, an exploratory method with which to identify interactions for further research. They illustrate this method using data from a US case-control study of myocardial infarction and stroke (1995-1999) carried out among 1,614 persons in Washington State who were genotyped for 32 SNPs on five genes in the renin-angiotensin system. PMID- 17082499 TI - Recovery of grasp versus reach in people with hemiparesis poststroke. AB - BACKGROUND AND OBJECTIVE: The authors recently found that grasping was not relatively more disrupted than reaching in people with acute hemiparesis. They now extend this work to the recovery of reach versus grasp. METHODS: Hemiparetic subjects were tested acutely, after 90 days, and then after 1 year poststroke, and a control group was evaluated once. Using kinematic techniques, subjects were studied performing reach and reach-to-grasp movements. The authors quantified 3 characteristics of performance for each movement: speed, accuracy, and efficiency, where an efficient movement was defined as a movement directly to the target without extraneous or abnormally circuitous movements. To evaluate the relative deficits and recovery in reach versus grasp, performance measures were converted to z scores using control group means and standard deviations. RESULTS: The authors' results showed that, starting with small deficits in speed acutely, both reach speed and grasp speed improved over time. Deficits in accuracy were greater in the reach than the grasp acutely, and these deficits lessened such that by the 90-day time point, the relative accuracy of the 2 movements was the same. In contrast, deficits in efficiency were greater in the grasp than the reach acutely, and grasp efficiency did not recover. The majority of recovery in reaching and grasping occurred by the 90-day time point, with little change occurring between the 90-day and 1-year time points. CONCLUSIONS: The authors hypothesize that, in chronic hemiparesis, purposeful movements requiring distal control may be more impaired than purposeful movements requiring proximal control, not because of the initial lesion, but because, over the course of recovery, spared components of the descending motor systems may be able to compensate for the accuracy deficits in reaching (proximal control) but not the efficiency deficits in grasping (distal muscular control). PMID- 17082500 TI - A single injection of D-amphetamine facilitates improvements in motor training following a focal cortical infarct in squirrel monkeys. AB - BACKGROUND: There is growing interest in the use of D-amphetamine (D-AMPH) as a pharmacological treatment to supplement rehabilitative therapy following stroke. Based on the success of earlier animal models, several clinical studies have demonstrated beneficial effects of applying physical rehabilitation while stroke patients are under the influence of D-AMPH. To begin to understand the neural mechanisms underlying this promising adjuvant therapy, the authors examined the effects of a single pairing of D-AMPH and rehabilitative training on motor performance after cortical infarct in squirrel monkeys. METHODS: Microelectrode stimulation techniques were used to delineate hand movement areas in the primary motor cortex prior to delivering a unilateral infarct to the complete hand representation. Postinfarct recovery was assessed for 3 groups of monkeys: D-AMPH + training, saline + training, and spontaneous recovery (SR). Postinfarct training groups received 14 consecutive days of motor skill training on a reach and retrieval task. A single injection of D-AMPH (0.25 mg/kg) or saline was given only on the 1st day of training (postinfarct day 10). Monkeys in the SR group had only minimal exposure to the training task once per week to monitor recovery. RESULTS: The results show that a single coupling of D-AMPH + training initiated 10 days after cortical infarct facilitated the rate of recovery and improved performance (68% improvement from 1st day of training) beyond the level achieved by the monkeys in the saline + training group (27% improved from 1st day of training). CONCLUSIONS: D-AMPH is a potent modulator of behavioral recovery following an ischemic infarct in nonhuman primates. PMID- 17082501 TI - Motor learning and Parkinson disease: Refinement of movement velocity and endpoint excursion in a limits of stability balance task. AB - OBJECTIVE: To investigate the effects of practice on performance and retention of a balance task in persons with Parkinson disease (PD). METHODS: Ten persons with PD and 10 age and gender-matched healthy control subjects were tested on an anticipatory, static base of support, limits of stability (LOS) balance task on a force plate. The motor learning paradigm utilized for all subjects included an acquisition phase and retention tests at 24 h and 1 week after acquisition. A force plate was used for testing and to collect outcome measures including movement velocity (MVL), endpoint excursion (EPE), and directional control. Data were analyzed for differences between groups and change over time. RESULTS: Persons with PD demonstrated performance deficits relative to controls for MVL at all testing periods (P < 0.05), and initially for EPE (P < 0.05), but were able to maintain significant improvements through retention testing relative to baseline (P < 0.05). CONCLUSIONS: Persons with PD demonstrated unimpaired capacity for motor learning in a LOS balance task for MVL and EPE, although performance deficits remained for MVL. The results concur with previous motor learning research of upper extremity tasks by suggesting that individuals with mild to moderate PD exhibit a preserved ability to benefit from practice as a means of improving balance task performance. PMID- 17082502 TI - The scale for contraversive pushing: A reliability and validity study. AB - OBJECTIVE: Pushing toward the hemiplegic side can interfere with mobility training after stroke. This study estimated the internal consistency, interrater reliability, and validity of the Scale for Contraversive Pushing (SCP). METHODS: Twenty-six patients with recent stroke were diagnosed with pusher behavior (PB). Two testers, randomly selected from 3 other examiners, independently assessed each patient using the SCP on the same day within 3 days of admission for rehabilitation. Cohen kappa coefficient was used to determine the agreement between the clinical and SCP diagnosis. The interrater reliability of the scale was estimated by calculation of the intraclass correlation coefficient. Cronbach's alpha coefficient and Pearson's coefficients were used to estimate the internal consistency of the scale and correlations between the subscores and the total score. RESULTS: The agreement between SCP and clinical diagnosis was very low when the original cutoff criterion for SCP diagnosis was used but was almost perfect with a modified criterion. The interrater reliability was good to excellent with regard both to each sub-score and to the total score. The internal consistency was very high, along with correlations between subscores and total score of the scale. CONCLUSIONS: The results provide support for use of the SCP based on its reliability and validity using a modified cutoff criterion to make a diagnosis of PB. PMID- 17082503 TI - Bilateral training does not facilitate performance of copying tasks in poststroke hemiplegia. AB - OBJECTIVE: Previous studies in rehabilitation in poststroke hemiplegia have suggested that brief periods of bilateral training improve subsequent unilateral performance of the paretic upper limb. The bilateral training protocol involved task-specific practice using both upper limbs simultaneously and homologously (such that they mirrored each other). This study aimed to determine the generalizability of the bilateral training phenomenon by replicating the work with different tasks, measures, and a different experimenter. METHODS: Five single-case experiments were performed using a within-participant multiple baseline design. Participants with post-stroke hemiplegia underwent 25 to 40 daily training sessions involving repetitive practice of 3 copying tasks. Pen movements were recorded on a digitizing pad. During the baseline phase, participants performed tasks unilaterally with their hemiplegic limb. Bilateral therapy was then applied in a staggered fashion to each task in turn. Quality of movement was measured in terms of temporal performance (jerkiness, speed, task duration), spatial performance (accuracy of copy), and arm posture (pen tilt). RESULTS: Movement quality of the hemiplegic limb was not improved during or subsequent to regular bilateral training. CONCLUSIONS: The training phenomenon observed in previous work was not replicated when participants with hemiplegia practiced copying tasks bilaterally. PMID- 17082504 TI - Evolution of subjective visual vertical perturbation after stroke. AB - OBJECTIVE: The perception of visual verticality is often perturbed after stroke and might be an underlying component of imbalance. The aim of this study was to describe the evolution of visual vertical (VV) perturbation and to investigate the factors affecting it. METHODS: Thirty patients with hemiplegia after a single hemispheric stroke (17 left lesioned [LL] and 13 right lesioned [RL]) were studied. Visual verticality was tested within 45 days of stroke, and then at 3 and 6 months. Subjects sat in a dark room and adjusted a luminous rod to the vertical position. The differences between patients' adjustments and vertical were calculated. The effects on VV evolution of the side, size, type, and location of the lesion were tested. RESULTS: Sixty percent of the recent stroke patients had an initial inaccurate perception of verticality, and 39% of these patients recovered during the 1st 3 months after stroke. The evolution of VV tilt depended on the side of the lesion (P = 0.01), with better recovery in LL patients. None of the other factors studied affected VV normalization. CONCLUSIONS: The poorer recovery of vertical perception after right-side stroke might be due to the predominant role of the right hemisphere in spatial cognition, and might be involved in the poorer recovery of balance after stroke in RL patients. PMID- 17082505 TI - Predictive value of ischemic lesion volume assessed with magnetic resonance imaging for neurological deficits and functional outcome poststroke: A critical review of the literature. AB - OBJECTIVE: Ischemic lesion volume is assumed to be an important predictor of poststroke neurological deficits and functional outcome. This critical review examines the methodological quality of MRI studies and the predictive value of hemispheric infarct volume for neurological deficits (at body function level) and functional outcome (at activities level). METHODS: Using Medline, PiCarta, and Embase to identify studies, 13 of the 747 identified studies met the authors' inclusion criteria. Subsequently, studies were tested for adherence to the key methodological criteria for internal, statistical, and external validity. Each criterion was weighted binary, and studies with 6 points or more were judged to be valid for assessing the predictive value of MRI for outcome. RESULTS: The 13 included studies had several methodological weaknesses with respect to internal validity, and none of them took lesion location into account. Only a few used outcome measures according to the International Classification of Functioning, Disability and Health and followed patients beyond 6 months. Correlation coefficients between MRI lesion volume and outcomes were higher for outcomes defined at body function level (National Institutes of Health Stroke Scale; median 0.67; range: 0.57-0.91) than for those defined at the level of activities (Barthel Index; median -0.49; range: -0.33 to -0.74). CONCLUSIONS: Methodological shortcomings of most studies confound the prognostic value of MRI in predicting stroke outcome, and few studies have focused on functional outcome. Future studies should investigate the added value of MRI volume over clinical neurological variables in predicting functional outcome beyond 6 months poststroke. PMID- 17082506 TI - A strategy for computer-assisted mental practice in stroke rehabilitation. AB - OBJECTIVE: To investigate the technical and clinical viability of using computer facilitated mental practice in the rehabilitation of upper-limb hemiparesis following stroke. DESIGN: A single-case study. SETTING: Academic-affiliated rehabilitation center. PARTICIPANT: A 46-year-old man with stable motor deficit of the upper right limb following subcortical ischemic stroke. INTERVENTION: Three computer-enhanced mental practice sessions per week at the rehabilitation center, in addition to usual physical therapy. A custom-made virtual reality system equipped with arm-tracking sensors was used to guide mental practice. The system was designed to superimpose over the (unseen) paretic arm a virtual reconstruction of the movement registered from the nonparetic arm. The laboratory intervention was followed by a 1-month home-rehabilitation program, making use of a portable display device. MAIN OUTCOME MEASURES: Pretreatment and posttreatment clinical assessment measures were the upper-extremity scale of the Fugl-Meyer Assessment of Sensorimotor Impairment and the Action Research Arm Test. Performance of the affected arm was evaluated using the healthy arm as the control condition. RESULTS: The patient's paretic limb improved after the first phase of intervention, with modest increases after home rehabilitation, as indicated by functional assessment scores and sensors data. CONCLUSION: Results suggest that technology-supported mental training is a feasible and potentially effective approach for improving motor skills after stroke. PMID- 17082507 TI - Neural internet: Web surfing with brain potentials for the completely paralyzed. AB - Neural Internet is a new technological advancement in brain-computer interface research, which enables locked-in patients to operate a Web browser directly with their brain potentials. Neural Internet was successfully tested with a locked-in patient diagnosed with amyotrophic lateral sclerosis rendering him the first paralyzed person to surf the Internet solely by regulating his electrical brain activity. The functioning of Neural Internet and its clinical implications for motor-impaired patients are highlighted. PMID- 17082510 TI - Carboplatin/cyclophosphamide or carboplatin/paclitaxel in elderly patients with advanced ovarian cancer? Analysis of two consecutive trials from the Groupe d'Investigateurs Nationaux pour l'Etude des Cancers Ovariens. AB - BACKGROUND: To determine the feasibility of two chemotherapy regimens in elderly patients with advanced ovarian carcinoma (AOC). PATIENTS AND METHODS: Eighty three patients >or=70 years were previously enrolled in a trial evaluating carboplatin and cyclophosphamide (CC). On the basis of identical eligibility criteria, 75 further patients were enrolled in a trial evaluating carboplatin and paclitaxel (Taxol) (CP). The primary end point of these studies was the feasibility of six courses of chemotherapy. Comprehensive geriatric assessment (CGA) parameters were assessed in terms of prognostic factors. RESULTS: More patients in the CC group presented with performance status of two or more, depression symptoms, use of co-medications, hypoalbuminemia, abnormal Mini-Mental Status score, or sub-optimal surgery. Both regimens appeared feasible: 75.6% in the CC group and 68.1% in the CP group completed six courses. CC and CP groups had similar overall survival (OS). Independent prognostic factors of poorer OS were the following: increasing age (P = 0.013), depression symptoms at baseline (P < 0.001), International Federation of Gynecology and Obstetrics stage IV (P = 0.001), and use of paclitaxel (P = 0.025). CONCLUSION: As this is a non randomised retrospective review of two consecutive studies, no firm conclusion can be drawn. It seems, however, that in elderly patients with AOC the use of paclitaxel results in more toxicity. CGA parameters and particularly emotional disorders might help to determine a priori the risk/benefit ratio of chemotherapy in this patient population. PMID- 17082512 TI - Assessment of renal toxicity and osteonecrosis of the jaws in patients receiving zoledronic acid for bone metastasis. AB - BACKGROUND: Bisphosphonates (BP) decrease the incidence of skeletal related events among cancer patients with bone metastases from solid tumors and multiple myeloma. Renal safety and osteonecrosis of the jaws (ONJ) are two major concerns of toxicity. Information about safety of using BP beyond 2 years is scarce. MATERIALS AND METHODS: Patients receiving zoledronic acid (ZA) at the time of the study were reviewed. Serum creatinine levels (SCL) were collected at three different moments: before the start of BP (baseline), at the time of analysis (final), and the highest SCL during the treatment (highest). Oral examination was carried out in every patient. Separated analysis was made for patients on BP for >2 years. Concomitant risk factors for both renal toxicity and ONJ were evaluated. RESULTS: Sixty-seven patients were included. Median time of BP was 22 months, with 22 patients receiving BP for >2 years. Median baseline and final values of SCL were 0.71 mg/dl and 0.70 mg/dl, respectively (P = 0.121). Median highest SCL during treatment was 0.82 mg/dl (P <0.0001). A notable increase in the SCL was observed in six of the 67 patients (9%), four of them receiving BP for >2 years (P = 0.085). ONJ was also diagnosed in six patients, four of them in the group of prolonged BP treatment. CONCLUSION: ZA showed to be safe with a low rate of reversible renal toxicity. Patients receiving BP should be monitored carefully for renal toxicity and ONJ, especially those with exposure to BP beyond 2 years. PMID- 17082511 TI - Epidermal growth factor receptor gene copy number and protein level are not associated with outcome of non-small-cell lung cancer patients treated with chemotherapy. AB - BACKGROUND: Survival benefit of non-small-cell lung cancer (NSCLC) patients treated with epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors is predicted by high EGFR gene copy number and by strong EGFR protein expression. Clinical relevance of these features in patients treated with chemotherapy has not been reported. PATIENTS AND METHODS: This study included 82 NSCLC patients treated with chemotherapy. There were 45% of females, 6% of never smokers and 45% of patients diagnosed with adenocarcinoma. EGFR gene copy number was evaluated by fluorescence in situ hybridization and EGFR protein level by immunohistochemistry. RESULTS: High EGFR gene copy number and protein level were found in 33% and 71% of patients, respectively. Both markers were significantly associated (P = 0.01). For objective response and disease control, there was no difference between patients defined as negative or positive for both EGFR gene copy number (P = 0.39 and P = 1.00, respectively) and for EGFR protein (P = 1.00 and P = 0.80, respectively). There were no differences in progression-free and overall survival according to EGFR gene copy number (P = 0.76 and P = 0.82, respectively) and protein level (P = 0.67 and P = 0.62, respectively). CONCLUSION: In chemotherapy-treated NSCLC patients, EGFR gene copy number was positively associated with protein level but none of the features were predictive for either treatment response or survival. PMID- 17082513 TI - Feasibility, toxicity and quality of life of first-line chemotherapy with platinum/paclitaxel in elderly patients aged >or=70 years with advanced ovarian cancer--a study by the AGO OVAR Germany. AB - BACKGROUND: The purpose of the study was to evaluate first-line platinum/paclitaxel (Taxol) under phase III trial conditions in ovarian cancer (OC) patients aged >or=70 years. PATIENTS AND METHODS: Phase III results of 779 patients with OC International Federation of Gynecology and Obstetrics (FIGO) stage IIB/IV treated with cisplatin/paclitaxel versus carboplatin/paclitaxel were retrospectively analyzed according to feasibility, toxicity (National Cancer Institute Common Toxicity Criteria) and quality of life (QoL) [European Organization for Research and Treatment of Cancer QoL questionnaire (EORTC QLQ C30)] in patients aged <70 or >or=70 years. RESULTS: One hundred and three (13%) patients were aged >or=70 years. Patient characteristics (<70 versus >or=70 years) showed significant differences with regard to Eastern Cooperative Oncology Group performance status, residual disease and constitutional factors but not to FIGO stage, histology or grading. Elderly patients received 98%, 100% and 96% of the recommended paclitaxel, carboplatin and cisplatin dose, respectively, per cycle. Early discontinuation was more frequent in elderly, although QoL, nonhematological and hematological toxicity were comparable between elderly and younger patients, except for febrile neutropenia (5% versus <1%, P = 0.005). There were no significant differences with regard to cycle delays, dose reductions or the use of granulocyte colony-stimulating factor and antibiotics. CONCLUSION: Platinum/paclitaxel appeared to be feasible and tolerable in elderly patients under clinical trial conditions, but there seems to be a different investigators' estimation of toxicity and less intention to maintain trial treatment in elderly. PMID- 17082514 TI - Heme oxygenase-1 is not required for mouse regulatory T cell development and function. AB - CD4 regulatory T cells (Treg) ensure peripheral tolerance to self-antigens and limit the deleterious effects associated with inflammatory and immune responses by mechanisms that remain to be fully understood. The enzyme heme oxygenase-1 (HO 1), through its known anti-inflammatory activity, is a candidate for a functional role in Treg activity. We compared wild-type and heme oxygenase-1-deficient (hmox 1(-/-)) mice in order to assess the role of HO-1 in mouse Treg development and function under physiologic conditions. The frequency of CD25+ and Foxp3+ Treg was similar in hmox-1(-/-) and hmox-1(+/+) mice. More importantly, CD4+ CD25+ Treg purified from either hmox-1(-/-) or hmox-1(+/+) mice were equally efficient in controlling the proliferation in vitro and the expansion in vivo of CD4+ CD25- T cells, whether or not these responder cells expressed HO-1. In addition, induction of expression of HO-1 in vivo did not affect Treg suppressor function. As shown before, expression of HO-1 was higher in Treg than in naive T cells; however, naturally activated Foxp3- T cells displayed equal amount of HO-1 mRNA as Treg. Finally, we conclude that under physiological conditions in mice, Treg development, maintenance and function are independent of HO-1 activity. PMID- 17082515 TI - Taste reactivity and Fos expression in GAD1-EGFP transgenic mice. AB - The central distribution of QHCl-elicited Fos-like immunoreactivity (FLI) suggests the location of a brain stem circuit that controls the oral rejection response. Although many species display an oral rejection response to bitter stimuli, the distribution of FLI associated with this response has been investigated only in rats. Fos data are minimal for the mouse, a species of increasing importance, due to its use in molecular and transgenic studies and taste-evoked oromotor responses are also only incompletely described in these rodents. We investigated these questions in FVB/NJ mice and a related transgenic strain (FVB-Tg(GadGFP)4507) that expresses green fluorescent protein in a subset of GAD1-containing neurons. QHCl, sucrose, or water delivered through intraoral cannulae yielded behavioral profiles that clearly differentiated QHCl from sucrose. Similar to rat, the number of neurons expressing FLI in the medial third of the solitary nucleus was elevated following QHCl compared with the other stimuli. In mice expressing green fluorescent protein, there was a pronounced distribution of GABAergic neurons in the ventral half of the solitary nucleus. Approximately 15% of solitary neurons expressing Fos were GABAergic, but this proportion did not differ according to stimulus. PMID- 17082516 TI - Specific configuration of dendritic degeneration in pyramidal neurons of the medial prefrontal cortex induced by differing corticosteroid regimens. AB - We previously demonstrated that hypercorticalism induces pronounced volumetric reductions in the rat medial prefrontal cortex (mPFC) and that these structural changes correlate with deficits in executive function. By applying 3-dimensional analysis of Golgi-Cox-stained material, we now demonstrate that corticosteroids can exert differential effects on dendritic arborizations of pyramidal neurons in lamina II/III of the mPFC. Treatment with the glucocorticoid receptor-selective agonist dexamethasone and with the natural adrenosteroid, corticosterone (CORT), results in significant reductions in the total length of apical dendrites in the pyramidal neurons in lamina II/III of the anterior cingulate/prelimbic and infralimbic cortices. Interestingly, although these treatments do not affect the number of dendritic branches, they are associated with impoverished arborizations in their distal portions and, in CORT-treated animals, with increased branching in the middle portions of the apical dendritic tree. Deprivation of corticosteroids by adrenalectomy leads to decreases in total apical dendritic length and spine number, but in this case, dendritic impoverishment was restricted to the middle/proximal segments of the dendritic trees. None of the treatments influenced the architecture of the basal dendrites. These results add to our knowledge of the morphological substrates through which corticosteroids may disrupt mPFC-dependent behaviors. PMID- 17082517 TI - Carpal tunnel syndrome and its relation to occupation: a systematic literature review. AB - OBJECTIVES: To assess occupational risk factors for carpal tunnel syndrome (CTS), we conducted a systematic literature review. METHODS: We identified relevant primary research from two major reviews in the 1990s and supplemented this material by a systematic search of the MEDLINE and EMBASE biomedical databases from the start of the electronic record to 1 January 2005. Reports were obtained and their bibliographies checked for other relevant publications. From each paper, we abstracted a standardized set of information on study populations, exposure contrasts and estimates of effect. RESULTS: Altogether, we summarized 38 primary reports, with analyses based either on a comparison of job titles (22) or of physical activities in the job (13) or both (3). We found reasonable evidence that regular and prolonged use of hand-held vibratory tools increases the risk of CTS >2-fold and found substantial evidence for similar or even higher risks from prolonged and highly repetitious flexion and extension of the wrist, especially when allied with a forceful grip. The balance of evidence on keyboard and computer work did not indicate an important association with CTS. Discussion Although the papers that we considered had limitations, a substantial and coherent body of evidence supports preventive policies aimed at avoiding highly repetitive wrist-hand work. There is a case for extending social security compensation for CTS in the United Kingdom to cover work of this kind. PMID- 17082518 TI - The impact of educational interventions on primary health care workers' knowledge of occupational exposure to blood or body fluids. AB - AIM: To assess the impact of educational interventions on primary health care workers' knowledge of management of occupational exposure to blood or body fluids. METHODS: Cluster-randomized trial of educational interventions in two National Health Service board areas in Scotland. Medical and dental practices were randomized to four groups; Group A, a control group of practices where staff received no intervention, Group B practices where staff received a flow chart regarding the management of blood and body fluid exposures, Group C received an e mail alert containing the flow chart and Group D practices received an oral presentation of information in the flow chart. Staff knowledge was assessed on one occasion, following the educational intervention, using an anonymous postal questionnaire. RESULTS: Two hundred and fifteen medical and dental practices were approached and 114 practices participated (response rate 53%). A total of 1120 individual questionnaires were returned. Face to face training was the most effective intervention with four of five outcome measures showing better than expected knowledge. Seventy-seven percent of staff identified themselves as at risk of exposure to blood and body fluids. Twenty-one percent of staff believed they were not at risk of exposure to blood-borne viruses although potentially exposed and 16% of exposed staff had not been immunized against hepatitis B. Of the 856 'at risk' staff, 48% had not received training regarding blood-borne viruses. CONCLUSIONS: We found greater knowledge regarding management of exposures to blood and body fluids following face to face training than other educational interventions. There is a need for education of at risk primary health care workers. PMID- 17082519 TI - Occupational stress and strain in the Naval Service: 1999 and 2004. AB - BACKGROUND: Surveys of occupational stress were undertaken in 1999 and 2004. OBJECTIVE: To measure the prevalence of psychological strain and exposure to work related stressors in personnel. METHOD: A cross-sectional survey using a validated, reliable questionnaire was undertaken in 1999 and was replicated in 2004. The data were combined and models of stress-strain were developed for male and female personnel in the Royal Navy (RN) and males in the Royal Marines (RM). RESULTS: The response rates were 78% in 1999 and 67% in 2004. Strain prevalence was 32% in 1999 and 33.5% in 2004. Strain rates were higher in females than in males and in personnel serving at sea. Reduced organizational commitment was associated with strain in males and females. Dissatisfaction with the physical work environment was associated with strain in RN females and RM. Role conflict was associated with strain in all groups. Work-family conflict was associated with strain in males, but not in females. The models accounted for 20-30% of the variance in strain. CONCLUSION: The strain rate in the Naval Service appeared to be similar to that in the other service occupations such as the UK Police, as did the sex difference in strain. The strain rate and stressor exposure ratings were remarkably stable over the period. Role conflict was found to be a generic stressor associated with strain. PMID- 17082522 TI - Tennis elbow in primary care. PMID- 17082523 TI - Families with disabled children. PMID- 17082524 TI - Breast feeding and intelligence in children. PMID- 17082525 TI - Pulmonary artery catheters. PMID- 17082526 TI - Glaucoma in the developing world. PMID- 17082527 TI - Regulatory bodies can discipline expert witnesses. PMID- 17082529 TI - Scientists ask Libya to release medical workers facing death sentence. PMID- 17082528 TI - Expert witnesses should deliver services through the NHS. PMID- 17082530 TI - Italian court upholds ban on pre-implantation diagnosis. PMID- 17082531 TI - Sex change expert accused of rushing patients into surgery. PMID- 17082532 TI - New rules for homoeopathic remedies anger UK peers. PMID- 17082546 TI - Acne. PMID- 17082547 TI - A practical approach to timing cord clamping in resource poor settings. PMID- 17082548 TI - ABC of obesity. Strategies for preventing obesity. PMID- 17082549 TI - Success relies on winning hearts and minds. PMID- 17082551 TI - Independence is key to better regulation. PMID- 17082552 TI - Many questions remain unanswered. PMID- 17082553 TI - A new role for the royal colleges? PMID- 17082554 TI - Health in the Middle East: HIV prevention is hindered in the United Arab Emirates. PMID- 17082555 TI - The BMJ interview: Sir Liam Donaldson: avoiding the painful truth and scapegoats. PMID- 17082556 TI - The BMJ interview: Sir Liam Donaldson: the health services and the dream. PMID- 17082557 TI - Health in the Middle East: no strong link between depleted uranium and cancer. PMID- 17082558 TI - Health in the Middle East: psychological implications of Iraqi invasion. PMID- 17082559 TI - Health in the Middle East: making healthcare systems more responsive to women in Pakistan. PMID- 17082560 TI - Investigating iron status in microcytic anaemia: causes and management of iron deficient anaemia. PMID- 17082561 TI - Investigating iron status in microcytic anaemia: zincprotoporphyrin and soluble transferrin receptor have a role. PMID- 17082562 TI - Investigating iron status in microcytic anaemia: positive diagnosis of anaemia of chronic disease. PMID- 17082563 TI - Investigating iron status in microcytic anaemia: general practitioners could test for ferritin, etc, before referral. PMID- 17082564 TI - A toxicogenomic approach revealed hepatic gene expression changes mechanistically linked to drug-induced hemolytic anemia. AB - A variety of pharmaceutical compounds causes hemolytic anemia as a significant adverse effect and this toxicity restricts the clinical utility of these drugs. In this study, we applied microarray technology to investigate hepatic gene expression changes associated with drug-induced hemolytic anemia and to identify potential biomarker genes for this hematotoxicity. We treated female Sprague Dawley rats with two hemolytic anemia-inducing compounds: phenylhydrazine and phenacetin. Hepatic gene expression profiles were obtained using a whole-genome oligonucleotide microarray with pooled RNA samples from individual rats within each dose group and analyzed in comparison with hepatic histopathology, hematology, and blood chemistry data. We identified a small subset of genes that were commonly deregulated in all the severe hemolytic conditions, some of which were considered to be involved in hepatic events characteristic of hemolytic anemia, such as hemoglobin biosynthesis, heme metabolism, and phagocytosis. Among them, we selected six upregulated genes as putative biomarkers, and their expression changes from microarray measurements were confirmed by quantitative real-time PCR using RNAs from individual animals. They were Alas2, beta-glo, Eraf, Hmox1, Lgals3, and Rhced. Expression patterns of all these genes showed high negative and positive correlation against erythrocyte counts and total bilirubin levels in circulation, respectively, suggesting that these genes may be the potential biomarkers for hemolytic anemia. These findings indicate that drug induced hemolytic anemia may be detected based on hepatic changes in the expression of a subset of genes that are mechanistically linked to the hematotoxicity. PMID- 17082565 TI - An environmental quinoid polycyclic aromatic hydrocarbon, acenaphthenequinone, modulates cyclooxygenase-2 expression through reactive oxygen species generation and nuclear factor kappa B activation in A549 cells. AB - Diesel exhaust particles (DEPs) contain oxygen-containing polycyclic aromatic hydrocarbons (PAHs) called quinoid PAHs. Some quinoid PAHs generate free radicals as they undergo enzymatic and nonenzymatic redox cycling with their corresponding semiquinone radicals. Reactive oxygen species (ROS) produced by these reactions can cause severe oxidative stress connected with inflammatory processing. Although humans and animals are continuously exposed to these chemicals in the environment, little is known about which quinoid PAHs are active. In this study, we estimated the intracellular ROS production and nuclear factor kappa B (NF kappaB) translocation in A549 cells exposed to isomers of quinoid PAHs having two to four rings. We found that both acenaphthenequinone (AcQ) and 9,10 phenanthrenequinone (PQ) enhanced ROS generation and that AcQ translocated NF kappaB from the cytosol to the nucleus. However, PQ, which has been reported to induce apoptosis, did not influence NF-kappaB activation. In addition, AcQ induced cyclooxygenase-2 (COX-2) expression which is a key enzyme in the inflammatory processing involved in the activation of NF-kappaB. Upregulation of NF-kappaB and COX-2 expression by AcQ treatment was suppressed by the antioxidant N-acetylcysteine (NAC). These results provide that AcQ might play an important role in human lung inflammatory diseases as an air pollutant. PMID- 17082566 TI - Nucleic acid-sensing TLRs as modifiers of autoimmunity. AB - The immune system requires precise regulation of activating and inhibitory signals so that it can mount effective responses against pathogens while ensuring tolerance to self-components. Some of the most potent activation signals are triggered by innate immune molecules, particularly those in the TLR family. Recent studies have shown that engagement of TLRs plays a significant role in both innate and adaptive immunity. This review focuses on the ways that TLR function might contribute to the etiology of lupus-like syndromes in the context of an autoimmune-prone environment. By considering the sources, localization, and expression of both nucleic acids and the molecules that bind them, we discuss several ways that innate immunity can play a role in the development of systemic autoimmunity. PMID- 17082567 TI - Cutting edge: CD4 is the receptor for the tick saliva immunosuppressor, Salp15. AB - Salp15 is an Ixodes scapularis salivary protein that inhibits CD4+ T cell activation through the repression of TCR ligation-triggered calcium fluxes and IL 2 production. We show in this study that Salp15 binds specifically to the CD4 coreceptor on mammalian host T cells. Salp15 specifically associates through its C-terminal residues with the outermost two extracellular domains of CD4. Upon binding to CD4, Salp15 inhibits the subsequent TCR ligation-induced T cell signaling at the earliest steps including tyrosine phosphorylation of the Src kinase Lck, downstream effector proteins, and lipid raft reorganization. These results provide a molecular basis to understanding the immunosuppressive activity of Salp15 and its specificity for CD4+ T cells. PMID- 17082568 TI - Cutting edge: activation of murine TLR8 by a combination of imidazoquinoline immune response modifiers and polyT oligodeoxynucleotides. AB - Synthetic immune response modifiers (IRM) such as imidazoquinolines can selectively activate human TLR7 or TLR8. Although these endosomal TLRs are close relatives, TLR7-deficient mice are unresponsive to TLR8 agonist IRMs. Similarly, natural ssRNA cannot activate murine TLR8, leading to the belief that murine TLR8 is nonfunctional. In this study, we transfected HEK293 cells with murine TLR8 and NF-kappaB reporter constructs and stimulated them with combinations of IRM and oligodeoxynucleotides (ODNs). When stimulated with TLR7 or TLR8 agonists alone, no NF-kappaB response was observed. However, a combination of polyT ODN plus the TLR8 agonist activated NF-kappaB, whereas polyT ODN plus the TLR7 agonist did not activate. Primary mouse cells responded to the IRM/polyT ODN by secreting TNF. Cells from TLR7(-/-) and TLR9(-/-) mice responded to the IRM/polyT ODN combination, whereas MyD88(-/-) cells did not respond. In conclusion, this study demonstrates for the first time that mouse TLR8 is functional. PMID- 17082569 TI - Cutting edge: resistance to HIV-1 infection among African female sex workers is associated with inhibitory KIR in the absence of their HLA ligands. AB - NK cells are regulated in part by killer Ig-like receptors (KIR) that interact with HLA molecules on potential target cells. KIR and HLA loci are highly polymorphic and certain KIR/HLA combinations were found to protect against HIV disease progression. We show in this study that KIR/HLA interactions also influence resistance to HIV transmission. HIV-exposed but seronegative female sex workers in Abidjan, Cote d'Ivoire, frequently possessed inhibitory KIR genes in the absence of their cognate HLA genes: KIR2DL2/KIR2DL3 heterozygosity in the absence of HLA-C1 and KIR3DL1 homozygosity in the absence of HLA-Bw4. HIV seropositive female sex workers were characterized by corresponding inhibitory KIR/HLA pairings: KIR2DL3 homozygosity together with HLA-C1 and a trend toward KIR3DL1/HLA-Bw4 homozygosity. Absence of ligands for inhibitory KIR could lower the threshold for NK cell activation. In addition, exposed seronegatives more frequently possessed AB KIR genotypes, which contain more activating KIR. The data support an important role for NK cells and KIR/HLA interactions in antiviral immunity. PMID- 17082570 TI - Cutting edge: inherent and acquired resistance to radiation-induced apoptosis in B cells: a pivotal role for STAT3. AB - Radiation-induced apoptosis (RiA) is used therapeutically for tumor cell ablation as well as a tool to characterize hemopoietic cell lineages. We report that the peritoneal B-1 B cell subset is selectively resistant to RiA. Inherent radioresistance is not shared by splenic B-2 or B-1 cells. However, it is conferred upon B-2 cells by BCR crosslinking in the presence of IL-6 or IL-10. In vivo experiments with gene-targeted mice confirm that IL-6 and, to a lesser extent, IL-10 are the relevant stimuli that combine with BCR ligands to promote B 1 cell radioresistance. STAT3 promotes cell survival in response to selected growth factors, and is activated by combined BCR crosslinking and IL-6 (IL-10). Importantly, STAT3(-/-) B-1 cells become susceptible to irradiation, indicating that STAT3 activation by the BCR in the presence of IL costimuli account for the inherent radioresistance of peritoneal B-1 B cells. PMID- 17082571 TI - Cutting edge: the phosphoinositide 3-kinase p110 delta is critical for the function of CD4+CD25+Foxp3+ regulatory T cells. AB - CD4+CD25+Foxp3+ regulatory T cells (Tregs) contribute to the maintenance of peripheral tolerance by inhibiting the expansion and function of conventional T cells. Treg development and homeostasis are regulated by the Ag receptor, costimulatory receptors such as CD28 and CTLA-4, and cytokines such as IL-2, IL 10, and TGF-beta. Here we show that the proportions of Tregs in the spleen and lymph nodes of mice with inactive p110delta PI3K (p110deltaD910A/D910A) are reduced despite enhanced Treg selection in the thymus. p110deltaD910A/D910A CD4+CD25+Foxp3+ Tregs showed attenuated suppressor function in vitro and failed to secrete IL-10. In adoptive transfer experiments, p110deltaD910A/D910A T cells failed to protect against experimental colitis. The identification of p110delta as an intracellular signaling protein that regulates the activity of CD4+CD25+Foxp3+ Tregs may facilitate the further elucidation of the molecular mechanisms responsible for Treg-mediated suppression. PMID- 17082572 TI - Cyclophosphamide-induced type-1 diabetes in the NOD mouse is associated with a reduction of CD4+CD25+Foxp3+ regulatory T cells. AB - Regulatory T cells (Tregs) have been implicated as key players in immune tolerance as well as suppression of antitumor responses. The chemotherapeutic alkylating agent cyclophosphamide (CY) is widely used in the treatment of tumors and some autoimmune conditions. Although previous data has demonstrated that Tregs may be preferentially affected by CY, its relevance in promoting autoimmune conditions has not been addressed. The nonobese diabetic mouse spontaneously develops type-1 diabetes (T1D). We demonstrate in this study that CY targets CD4+CD25+Foxp3+ Tregs in vivo. CD4+CD25+ T cells isolated from CY-treated mice display reduced suppressive activity in vitro and increased expression of apoptotic markers. Although Treg numbers rapidly recovered to pretreatment levels in the peripheral lymphoid tissues, Tregs failed to recover proportionally within pancreatic infiltrates. T1D progression was effectively prevented by adoptive transfer of a small number of islet Ag-specific CD4+CD25+ Tregs to CY-treated recipients. Prevention of T1D was associated with reduced T cell activation and higher Treg proportions in the pancreas. We conclude that acceleration of T1D by CY is associated with a reduction in CD4+CD25+Foxp3+ Tregs and can be prevented by transfer of CD4+CD25+ Tregs. PMID- 17082573 TI - Coreceptor signal strength regulates positive selection but does not determine CD4/CD8 lineage choice in a physiologic in vivo model. AB - TCR signals drive thymocyte development, but it remains controversial what impact, if any, the intensity of those signals have on T cell differentiation in the thymus. In this study, we assess the impact of CD8 coreceptor signal strength on positive selection and CD4/CD8 lineage choice using novel gene knockin mice in which the endogenous CD8alpha gene has been re-engineered to encode the stronger signaling cytoplasmic tail of CD4, with the re-engineered CD8alpha gene referred to as CD8.4. We found that stronger signaling CD8.4 coreceptors specifically improved the efficiency of CD8-dependent positive selection and quantitatively increased the number of MHC class I (MHC-I)-specific thymocytes signaled to differentiate into CD8+ T cells, even for thymocytes expressing a single, transgenic TCR. Importantly, however, stronger signaling CD8.4 coreceptors did not alter the CD8 lineage choice of any MHC-I-specific thymocytes, even MHC-I specific thymocytes expressing the high-affinity F5 transgenic TCR. This study documents in a physiologic in vivo model that coreceptor signal strength alters TCR-signaling thresholds for positive selection and so is a major determinant of the CD4:CD8 ratio, but it does not influence CD4/CD8 lineage choice. PMID- 17082574 TI - Prime-boost with alternating DNA vaccines designed to engage different antigen presentation pathways generates high frequencies of peptide-specific CD8+ T cells. AB - The route for presentation of Ag to CD8+ or CD4+ T cells following DNA vaccination is critical for determining outcome, but the pathways involved are unclear. In this study, we compare two different DNA vaccine designs aimed to elicit CD8+ T cell responses against a specific peptide-epitope either by direct- or cross-presentation. Each carries sequences from tetanus toxin (TT) to provide essential CD4+ T cell help. In the first already proven design, the peptide epitope is fused to the N-terminal domain of fragment C from TT. This appears to act mainly by cross-presentation. In the second design, the peptide-epitope is encoded by a minigene, with induction of Th responses mediated by coexpression of a hybrid invariant chain molecule, incorporating a single determinant from TT (p30) in exchange for class II-associated invariant chain peptide. This design appears to act mainly via direct presentation from transfected APCs. Both vaccines mediated Th-dependent priming of CD8+ T cells in mice, but the kinetics and level of the responses differed markedly, consistent with engagement of distinct pathways of Ag presentation. Importantly, the vaccines could be combined in an alternating prime-boost regime, in either order, generating substantially expanded memory CD8+ T cells, with potent effector function. Taken together, these results demonstrate that vaccination protocols involving different modes of Ag presentation at prime and boost can significantly improve the effectiveness of immunization. PMID- 17082575 TI - Antibody responses to mycobacterial and self heat shock protein 65 in autoimmune arthritis: epitope specificity and implication in pathogenesis. AB - Many autoimmune diseases are believed to involve primarily T cell-mediated effector mechanisms. There is increasing realization, however, that Abs may also play a vital role in the propagation of T cell-driven disorders. In this study, on the rat adjuvant-induced arthritis (AA) model of human rheumatoid arthritis, we examined the characteristics of serum Ab response to mycobacterial heat shock protein (hsp) 65 (Bhsp65), self (rat) hsp65 (Rhsp65), and linear peptides spanning these two molecules. The AA-resistant WKY (RT.1(l)) rat responded to the heat-killed Mycobacterium tuberculosis immunization with a rapid burst of Abs to both Bhsp65 and Rhsp65. These Abs reacted with numerous peptide epitopes; however, this response was reduced to a few epitopes with time. On the contrary, the susceptible Lewis (RT.1(l)) rat developed a relatively lower Ab response to Bhsp65, and Abs to Rhsp65 did not appear until the recovery from the disease. The Ab response in Lewis rats diversified with progression of AA, and there was an intriguing overlap between the repertoire of Bhsp65-reactive B and T cells during the recovery phase of AA. Nonetheless, subsets of the repertoire of the late Abs in both rat strains became focused on the same epitope regions of Bhsp65 and Rhsp65. The functional relevance of these Abs was evident from the results showing that sera from recovery phase Lewis or WKY rats, but not that of naive rats, afforded protection against subsequent AA. These results are of significance in further understanding of the role of humoral immunity in the pathogenesis of autoimmune arthritis. PMID- 17082576 TI - Functional dichotomy in CD40 reciprocally regulates effector T cell functions. AB - Activation of T cells requires signals through Ag-specific TCR and costimulatory molecules such as CD40L. Although the use of defined tumor Ags for the induction of protective T cells met with limited success, the CD40-CD40L interaction that was proposed to induce antitumor T cells did not prevent tumor growth completely. Using a model for prostate tumor, a leading cause of tumor-induced mortality in men, we show that the failure is due to a novel functional dichotomy of CD40 whereby it self-limits its antitumor functions by inducing IL-10. IL-10 prevents the CD40-induced CTL and TNF-alpha and IL-12 production, Th1 skewing, and tumor regression. Priming mice with tumor lysate-pulsed IL-10-deficient dendritic cells (DCs) or wild-type DC plus anti-IL-10 Ab establishes antitumor memory T cells that can transfer the protection into syngenic nude mice. Infusion of Ag-pulsed IL-10-deficient but not wild-type DCs back into syngenic mice results in successful therapeutic autovaccination. Thus, we demonstrate the IL-10-sensitive antitumor T cell memory formulating a novel prophylactic and therapeutic principle. PMID- 17082577 TI - GATA-3 regulates the development and function of invariant NKT cells. AB - Although invariant NKT (iNKT) cells participate in many aspects of immune responses, the molecular mechanisms regulating their development, maturation, and activation are still poorly understood. GATA-3 is a T cell-specific transcription factor that is also expressed in iNKT cells. The critical role of GATA-3 in conventional alphabeta T cells has been well documented, but whether GATA-3 also regulates the development and function of iNKT cells is unknown. In the present study, we report that deficiency of GATA-3 results in cell-intrinsic defects in the thymic development and peripheral maturation of murine iNKT cells. In addition, GATA-3 is also required for survival, activation, and effector functions of this unique population of T cells. Our data also reveal a previously unidentified peripheral maturation step that is GATA-3 dependent. PMID- 17082578 TI - Immature CD4+CD8+ thymocytes and mature T cells regulate Nur77 distinctly in response to TCR stimulation. AB - The orphan steroid receptor, Nur77, is thought to be a central participant in events leading to TCR-mediated clonal deletion of immature thymocytes. Interestingly, although both immature and mature murine T cell populations rapidly up-regulate Nur77 after TCR stimulation, immature CD4+CD8+ thymocytes respond by undergoing apoptosis, whereas their mature descendants respond by dividing. To understand these developmental differences in susceptibility to the proapoptotic potential of Nur77, we compared its regulation and compartmentalization and show that mature, but not immature, T cells hyperphosphorylate Nur77 in response to TCR signals. Nur77 resides in the nucleus of immature CD4+CD8+ thymocytes throughout the course of its expression and is not found in either the organellar or cytoplasmic fractions. However, hyperphosphorylation of Nur77 in mature T cells, which is mediated by both the MAPK and PI3K/Akt pathways, shifts its localization from the nucleus to the cytoplasm. The failure of immature CD4+CD8+ thymocytes to hyperphosphorylate Nur77 in response to TCR stimulation may be due in part to decreased Akt activity at this developmental stage. PMID- 17082579 TI - Specific and redundant roles for NFAT transcription factors in the expression of mast cell-derived cytokines. AB - By virtue of their ability to express a plethora of biologically highly active mediators, mast cells (MC) are involved in both adaptive and innate immune responses. MC-derived Th2-type cytokines are thought to act as local amplifiers of Th2 reactions, including chronic inflammatory disorders such as allergic asthma, whereas MC-derived TNF-alpha is a critical initiator of antimicrobial defense. In this study, we demonstrate that the transcription factors NFATc1 and NFATc2 are part of a MC-specific signaling network that regulates the expression of TNF-alpha and IL-13, whereas NFATc3 is dispensable. Primary murine bone marrow derived MC from NFATc2(-/-) mice, activated by either ionomycin or IgE/Ag cross link, display a strong reduction in the production of these cytokines, compared with bone marrow-derived MC from wild-type mice. Detailed analyses of TNF-alpha and IL-13 expression using small interfering RNA-mediated knockdown reveals that both NFATc2 and NFATc1 are able to drive the expression of these cytokines, whereas neither degranulation nor the expression of IL-6 depends on NFAT activity. These results support the view that high NFAT activity is necessary for TNF-alpha and IL-13 promoter induction in MC, irrespective of whether NFATc2 or NFATc1 or a combination of both is present. PMID- 17082580 TI - Partial versus full allogeneic hemopoietic chimerization is a preferential means to inhibit type 1 diabetes as the latter induces generalized immunosuppression. AB - In both humans and NOD mice, particular combinations of MHC genes provide the primary risk factor for development of the autoreactive T cell responses causing type 1 diabetes (T1D). Conversely, other MHC variants can confer dominant T1D resistance, and previous studies in NOD mice have shown their expression on hemopoietically derived APC is sufficient to induce disease protection. Although allogeneic hemopoietic chimerization can clearly provide a means for blocking T1D development, its clinical use for this purpose has been obviated by a requirement to precondition the host with what would be a lethal irradiation dose if bone marrow engraftment is not successful. There have been reports in which T1D protective allogeneic hemopoietic chimerization was established in NOD mice that were preconditioned by protocols not including a lethal dose of irradiation. In most of these studies, virtually all the hemopoietic cells in the NOD recipients eventually converted to donor type. We now report that a concern about such full allogeneic chimeras is that they are severely immunocompromised potentially because their T cells are positively selected in the thymus by MHC molecules differing from those expressed by the APC available in the periphery to activate T cell effector functions. However, this undesirable side effect of generalized immunosuppression is obviated by a new protocol that establishes without a lethal preconditioning component, a stable state of mixed allogeneic hemopoietic chimerism sufficient to inhibit T1D development and also induce donor-specific tolerance in NOD recipients. PMID- 17082581 TI - Death of CD4+ T cells from lymph nodes during primary SIVmac251 infection predicts the rate of AIDS progression. AB - Immunological and virological events that occur during the earliest stages of SIV infection are now considered to have a major impact on subsequent disease progression. In the present study, we demonstrate a clear correlation between progression to AIDS and the rate of in vitro CD4+ (but not CD8+) T cell death in lymph nodes. The dying CD4+ T cells were effector memory T cells, which are critical for the immune response to pathogens. However, there was no correlation between the rate of the viral replication within lymph nodes and the extent of Fas ligand-mediated death, despite the increased sensitivity of CD4+ T cells to death in response to recombinant human Fas ligand. CD4+ T cell death was caspase and apoptosis-inducing factor independent but was clearly associated with mitochondrion damage. Interestingly, higher expression levels of the active form of Bak, a proapoptotic molecule involved in mitochondrial membrane permeabilization, were observed in SIV-infected macaques progressing more rapidly to AIDS. Finally, we demonstrated that the strain of SIV we used requires CCR5 and BOB/GRP15 molecules as coreceptors and caused death of unstimulated noncycling primary CD4+ T cells. Altogether, these results demonstrate that CD4+ T cell death occurring early after SIV infection is a crucial determinant of progression to AIDS and that it is mediated by the intrinsic death pathway. PMID- 17082582 TI - Glutamate released by dendritic cells as a novel modulator of T cell activation. AB - Adaptive immune responses begin after productive immunosynaptic contacts formation established in secondary lymphoid organs by dendritic cells (DC) presenting the Ag to T lymphocytes. Despite its resemblance to the neurosynapse, the participation of soluble small nonpeptidic mediators in the intercellular cross-talk taking place during T cell-DC interactions remains poorly studied. In this study, we show that human DC undergoing maturation and in contact with T cells release significant amounts of glutamate, which is the main excitatory neurotransmitter in mammalians. The release of glutamate is nonvesicular and mediated by the DC-expressed Xc- cystine/glutamate antiporter. DC-derived glutamate stimulating the constitutively expressed metabotropic glutamate receptor 5 impairs T cell activation. However, after productive Ag presentation, metabotropic glutamate receptor 1 is expressed in T cells to mediate enhanced T cell proliferation and secretion of Th1 and proinflammatory cytokines. These data suggest that, during T cell-DC interaction, glutamate is a novel and highly effective regulator in the initiation of T cell-mediated immune responses. PMID- 17082583 TI - Quantification of repertoire diversity of influenza-specific epitopes with predominant public or private TCR usage. AB - The H-2Db-restricted CD8 T cell immune response to influenza A is directed at two well-described epitopes, nucleoprotein 366 (NP366) and acid polymerase 224 (PA224). The responses to the two epitopes are very different. The epitope NP366 specific response is dominated by TCR clonotypes that are public (shared by most mice), whereas the epitope PA224-specific response is private (unique within each infected animal). In addition to being public, the NP366-specific response is dominated by a few clonotypes, when T cell clonotypes expressing the Vbeta8.3 element are analyzed. Herein, we show that this response is similarly public when the NP366+Vbeta4+ CD8 T cell response is analyzed. Furthermore, to determine whether these features resulted in differences in total TCR diversity in the NP366+ and PA224+ responses, we quantified the number of different CD8 T clonotypes responding to each epitope. We calculated that 50-550 clonotypes recognized each epitope in individual mice. Thus, although the character of the response to the two epitopes appeared to be different (private and diverse vs public and dominated by a few clonotypes), similar numbers of precursor cells responded to both epitopes and this number was of similar magnitude to that previously reported for other viral CD8 T cell epitopes. Therefore, even in CD8 T cell responses that appear to be oligoclonotypic, the total response is highly diverse. PMID- 17082584 TI - CXC chemokine ligand 13 and CC chemokine ligand 19 cooperatively render resistance to apoptosis in B cell lineage acute and chronic lymphocytic leukemia CD23+CD5+ B cells. AB - CXCL13/CXCR5 and CCL19/CCR7 play a quite important role in normal physiological conditions, but the functions of both chemokine/receptor pairs in pathophysiological events are not well-investigated. We have investigated expression and functions of CXCL13/CXCR5 and CCL19/CCR7 in CD23+CD5+ and CD23+CD5 B cells from cord blood (CB) and patients with B cell lineage acute or chronic lymphocytic leukemia (B-ALL or B-CLL). CXCR5 and CCR7 are selectively expressed on B-ALL, B-CLL, and CB CD23+CD5+ B cells at high frequency, but not on CD23+CD5- B cells. Although no significant chemotactic responsiveness was observed, CXCL13 and CCL19 cooperatively induce significant resistance to TNF-alpha-mediated apoptosis in B-ALL and B-CLL CD23+CD5+ B cells, but not in the cells from CB. B ALL and B-CLL CD23+CD5+ B cells express elevated levels of paternally expressed gene 10 (PEG10). CXCL13 and CCL19 together significantly up-regulate PEG10 expression in the same cells. We have found that CXCL13 and CCL19 together by means of activation of CXCR5 and CCR7 up-regulate PEG10 expression and function, subsequently stabilize caspase-3 and caspase-8 in B-ALL and B-CLL CD23+CD5+ B cells, and further rescue the cells from TNF-alpha-mediated apoptosis. Therefore, we suggest that normal lymphocytes, especially naive B and T cells, use CXCL13/CXCR5 and CCL19/CCR7 for migration, homing, maturation, and cell homeostasis as well as secondary lymphoid tissues organogenesis. In addition, certain malignant cells take advantages of CXCL13/CXCR5 and CCL19/CCR7 for infiltration, resistance to apoptosis, and inappropriate proliferation. PMID- 17082585 TI - E2-2 regulates the expansion of pro-B cells and follicular versus marginal zone decisions. AB - The E-proteins E2A, HeLa E-box binding protein, and E2-2 constitute a class of basic helix-loop-helix transcription factors that differentially affect B cell development. E2A is by far the most investigated and appears to operate at several levels during B cell ontogeny. Less is known concerning the role of the other E-proteins. To address the role of E2-2, we have performed transfers of fetal liver (FL) cells into irradiated Rag-deficient mice. Although the transfer of E2-2-deficient cells alone can reconstitute all B cell subpopulations, albeit with a moderate reduction in cellularity, E2-2-deficient cells have a disadvantage when transferred together with wild-type cells. Cultivation of E2-2( /-) day 14.5 FL cells on stromal cells and IL-7 revealed a reduced frequency of responding B cell progenitors despite normal IL-7Ralpha surface expression. Real time PCR analysis revealed that E2-2 mRNA expression is high at the pro-B cell stage and drops sharply at the pre-B cell stage, consistent with a role for E2-2 in pro-B cells. In contrast, E2A mRNA was most abundant in pre-B cells. Analysis of the peripheral repertoire revealed that mice reconstituted with E2-2(-/-) FL cells had an increased proportion of marginal zone (MZ) B cells. Interestingly, E2-2 mRNA was elevated approximately 2-fold (p < 0.01) in follicular compared with MZ B cells. Although E2A mRNA showed a similar tendency, the difference was not significant. Collectively, our findings indicate that E2-2 is required for optimal expansion of pro-B cells, and also influences the follicular vs MZ decision. PMID- 17082586 TI - Human bone marrow: a reservoir for "enhanced effector memory" CD8+ T cells with potent recall function. AB - The role of human bone marrow (BM) CD8+ T cells in the immune response to viral Ags is poorly defined. We report here the identification and characterization of a functionally enhanced effector memory CD8+ T cell population (TEM) in the BM of patients undergoing total joint replacement for osteoarthritis. These BM-derived TEM differ strikingly from correlate cells in peripheral blood (PB), expressing elevated levels of CD27, HLA-DR, CD38, CD69, and unique patterns of chemokine receptors. Interestingly, while BM TEM have low levels of resting perforin and granzyme B, these molecules evidence profound up-regulation in response to TCR stimulation resulting in enhanced cytotoxic potential. Moreover, compared with the TEM subset in PB, BM CD8+ TEM cells demonstrate a more vigorous recall response to pooled viral Ags. Our results reveal that human BM serves as a repository for viral Ag-specific TEM with great therapeutic potential in vaccine development. PMID- 17082587 TI - CD8 T cell recall responses are regulated by the tissue tropism of the memory cell and pathogen. AB - Whether memory CD8 T cells can be reactivated in nonlymphoid tissues is unclear. Using mice lacking the spleen, lymph nodes, or both, we show that the secondary T cell response, but not homeostatic maintenance of memory cells, required lymphoid tissue. Whereas primary and secondary CD8 T cell responses to vesicular stomatitis virus infection were lymph node dependent, responses to Listeria monocytogenes infection were driven primarily in the spleen. Memory cell subset reactivation was also regulated by location of the responding population and the pathogen. Thus, CD62Llow effector memory T cells (TEM) cells responded nearly as well as CD62Lhigh central memory T cells (TCM) and TCM cells after L. monocytogenes infection, and both subsets generated equivalent populations of secondary memory cells. In contrast, TCM cells, but not TEM cells, mounted a robust response to vesicular stomatitis virus infection. TCM and TEM cells also required lymphoid tissue to mount recall responses, and the bone marrow did not contribute significantly to the response of either subset. Our findings indicated that characteristics of the infectious agent and the migratory preferences of memory cells dictated the secondary lymphoid tissue requirement for the recall response to infection. PMID- 17082588 TI - CD4+ T cells in the absence of the CD8+ cytotoxic T cells are critical and sufficient for NKT cell-dependent tumor rejection. AB - NKT cells perform crucial roles in tumor surveillance, functioning as regulators of early host response. In this study, we have assessed the effects of NKT activation at the time of tumor Ag immunization, and have evaluated the contributions of CD4+ and CD8+ T cells in tumor rejection during adaptive immune response against live tumor cells. Our data indicate that CD4+ T cells play critical roles, not only in assisting CTL, but also in the orchestration of host response against the tumor. The CD4+ T cells were found to reject the transplanted tumor cells very efficiently under conditions in which the CTLs were removed either genetically, or via the action of anti-CD8 Ab in mice that had been immunized with tumor extracts and alpha-galactosylceramide. Immunization resulted in an NKT cell-dependent antitumor adaptive immune response, which was associated with both CD4+ T cells and cytokine IFN-gamma. PMID- 17082589 TI - Hepatitis C virus (HCV) core protein-induced, monocyte-mediated mechanisms of reduced IFN-alpha and plasmacytoid dendritic cell loss in chronic HCV infection. AB - IFN-alpha production by plasmacytoid dendritic cells (PDCs) is critical in antiviral immunity. In the present study, we evaluated the IFN-alpha-producing capacity of PDCs of patients with chronic hepatitis C virus (HCV) infection in treatment-naive, sustained responder, and nonresponder patients. IFN-alpha production was tested in PBMCs or isolated PDCs after TLR9 stimulation. Treatment naive patients with chronic HCV infection had reduced frequency of circulating PDCs due to increased apoptosis and showed diminished IFN-alpha production after stimulation with TLR9 ligands. These PDC defects correlated with the presence of HCV and were in contrast with normal PDC functions of sustained responders. HCV core protein, which was detectable in the plasma of infected patients, reduced TLR9-triggered IFN-alpha and increased TNF-alpha and IL-10 production in PBMCs but not in isolated PDCs, suggesting HCV core induced PDC defects. Indeed, addition of rTNF-alpha and IL-10 induced apoptosis and inhibited IFN-alpha production in PDCs. Neutralization of TNF-alpha and/or IL-10 prevented HCV core induced inhibition of IFN-alpha production. We identified CD14+ monocytes as the source of TNF-alpha and IL-10 in the HCV core-induced inhibition of PDC IFN-alpha production. Anti-TLR2-, not anti-TLR4-, blocking Ab prevented the HCV core induced inhibition of IFN-alpha production. In conclusion, our results suggest that HCV interferes with antiviral immunity through TLR2-mediated monocyte activation triggered by the HCV core protein to induce cytokines that in turn lead to PDC apoptosis and inhibit IFN-alpha production. These mechanisms are likely to contribute to HCV viral escape from immune responses. PMID- 17082590 TI - Melan-A/MART-1-specific CD4 T cells in melanoma patients: identification of new epitopes and ex vivo visualization of specific T cells by MHC class II tetramers. AB - Over the past decade, many efforts have been made to identify MHC class II restricted epitopes from different tumor-associated Ags. Melan-A/MART-1(26-35) parental or Melan-A/MART-1(26-35(A27L)) analog epitopes have been widely used in melanoma immunotherapy to induce and boost CTL responses, but only one Th epitope is currently known (Melan-A51-73, DRB1*0401 restricted). In this study, we describe two novel Melan-A/MART-1-derived sequences recognized by CD4 T cells from melanoma patients. These epitopes can be mimicked by peptides Melan-A27-40 presented by HLA-DRB1*0101 and HLA-DRB1*0102 and Melan-A25-36 presented by HLA DQB1*0602 and HLA-DRB1*0301. CD4 T cell clones specific for these epitopes recognize Melan-A/MART-1+ tumor cells and Melan-A/MART-1-transduced EBV-B cells and recognition is reduced by inhibitors of the MHC class II presentation pathway. This suggests that the epitopes are naturally processed and presented by EBV-B cells and melanoma cells. Moreover, Melan-A-specific Abs could be detected in the serum of patients with measurable CD4 T cell responses specific for Melan A/MART-1. Interestingly, even the short Melan-A/MART-1(26-35(A27L)) peptide was recognized by CD4 T cells from HLA-DQ6+ and HLA-DR3+ melanoma patients. Using Melan-A/MART-1(25-36)/DQ6 tetramers, we could detect Ag-specific CD4 T cells directly ex vivo in circulating lymphocytes of a melanoma patient. Together, these results provide the basis for monitoring of naturally occurring and vaccine induced Melan-A/MART-1-specific CD4 T cell responses, allowing precise and ex vivo characterization of responding T cells. PMID- 17082591 TI - T regulatory and primed uncommitted CD4 T cells express CD73, which suppresses effector CD4 T cells by converting 5'-adenosine monophosphate to adenosine. AB - CD73 (5'-ectonucleotidase) is expressed by two distinct mouse CD4 T cell populations: CD25+ (FoxP3+) T regulatory (Treg) cells that suppress T cell proliferation but do not secrete IL-2, and CD25- uncommitted primed precursor Th (Thpp) cells that secrete IL-2 but do not suppress in standard Treg suppressor assays. CD73 on both Treg and Thpp cells converted extracellular 5'-AMP to adenosine. Adenosine suppressed proliferation and cytokine secretion of Th1 and Th2 effector cells, even when target cells were activated by anti-CD3 and anti CD28. This represents an additional suppressive mechanism of Treg cells and a previously unrecognized suppressive activity of Thpp cells. Infiltration of either Treg or Thpp cells at inflammatory sites could potentially convert 5'-AMP generated by neutrophils or dying cells into the anti-inflammatory mediator adenosine, thus dampening excessive immune reactions. PMID- 17082592 TI - Activin A functions as a Th2 cytokine in the promotion of the alternative activation of macrophages. AB - Activin A, a member of the TGF-beta superfamily, is a pluripotent growth and differentiation factor. In this study, we report that murine Th cells produce activin A upon activation. Activin activity in the cultured CD4+ T cells was induced by anti-CD3 cross-linking. Activin betaA mRNA level was increased in response to activation, indicating that activin production in CD4+ T cells is regulated at the mRNA level. Activin production was detected exclusively in CD4+CD25- T cells, but not in CD4+CD25+ regulatory T cells. When CD4+ T cells were differentiated into Th cell subsets, higher activin secretion was detected when cultured under Th2-skewing conditions. The mRNA level of activin betaA was abundant in Th2, but not in Th1 cells. Furthermore, secretion of activin was significantly higher in activated Th2 clones than in Th1 clones. The activin betaA-proximal promoter contains a binding site for c-Maf, a Th2-specific transcriptional factor, at close proximity with an NF-AT binding site. c-Maf was able to synergize with NF-AT to transactivate activin betaA gene, and both factors are implicated in activin betaA transcription in Th2 cells. Activin A induced macrophages to express arginase-1 (M-2 phenotype), whereas it inhibited inducible NO synthase expression (M-1 phenotype) induced by IFN-gamma. Taken together, these observations suggest that activin A is a novel Th2 cytokine that promotes differentiation of macrophages toward the M-2 phenotype. PMID- 17082593 TI - The wild-type sequence (wt) p53(25-35) peptide induces HLA-DR7 and HLA-DR11 restricted CD4+ Th cells capable of enhancing the ex vivo expansion and function of anti-wt p53(264-272) peptide CD8+ T cells. AB - Tumor peptide-based vaccines are more effective when they include tumor-specific Th cell-defined as well as CTL-defined peptides. Presently, two overlapping wild type sequences (wt) p53 helper peptides, p53(108-122) and p53(110-124), have been identified as HLA-DR1- and/or HLA-DR4-restricted epitopes. These HLA-DR alleles are expressed by approximately 35% of subjects with cancer. To identify Th cell defined wt p53 peptides suitable for use on the remaining subject population, a dendritic cell (DC)-based coculture system was developed. CD4+ T cells isolated from PBMC obtained from HLA-DR4- normal donors were stimulated ex vivo with autologous DC transfected with wt p53 or mutant p53 cDNA. Reactivity of T cells was tested in ELISPOT IFN-gamma assays against DC pulsed individually with a panel of algorithm-predicted, multiple HLA-DR-binding wt p53 peptides. The wt p53(25-35) peptide was identified as capable of inducing and being recognized by CD4+ T cells in association, at a minimum, with HLA-DR7 and -DR11 molecules, each of which is expressed by approximately 15% of the population. In addition, the presence of anti-p53(25-35) CD4+ Th cells was shown to enhance the in vitro generation/expansion of HLA-A2-restricted, anti-wt p53(264-272) CD8+ T cells, which from one donor were initially "nonresponsive" to the wt p53(264-272) peptide. The wt p53(25-35) peptide has attributes of a naturally presented Th cell-defined peptide, which could be incorporated into antitumor vaccines applicable to a broader population of subjects for whom a wt p53 helper peptide is presently unavailable, as well as used for monitoring anti-p53 Th cell activity in cancer subjects receiving p53-based immunotherapy. PMID- 17082594 TI - TCR alpha genes direct MHC restriction in the potent human T cell response to a class I-bound viral epitope. AB - The underlying generic properties of alphabeta TCRs that control MHC restriction remain largely unresolved. To investigate MHC restriction, we have examined the CTL response to a viral epitope that binds promiscuously to two human leukocyte Ags (HLAs) that differ by a single amino acid at position 156. Individuals expressing either HLA-B*3501 (156Leucine) or HLA-B*3508 (156Arginine) showed a potent CTL response to the 407HPVGEADYFEY417 epitope from EBV. Interestingly, the response was characterized by highly restricted TCR beta-chain usage in both HLA B*3501+ and HLA-B*3508+ individuals; however, this conserved TRBV9+ beta-chain was associated with distinct TCR alpha-chains depending upon the HLA-B*35 allele expressed by the virus-exposed host. Functional assays confirmed that TCR alpha chain usage determined the HLA restriction of the CTLs. Structural studies revealed significant differences in the mobility of the peptide when bound to HLA B*3501 or HLA-B*3508. In HLA-B*3501, the bulged section of the peptide was disordered, whereas in HLA-B*3508 the bulged epitope adopted an ordered conformation. Collectively, these data demonstrate not only that mobile MHC-bound peptides can be highly immunogenic but can also stimulate an extremely biased TCR repertoire. In addition, TCR alpha-chain usage is shown to play a critical role in controlling MHC restriction between closely related allomorphs. PMID- 17082595 TI - Fc receptor homolog 3 is a novel immunoregulatory marker of marginal zone and B1 B cells. AB - Two members of the recently identified FcR homolog (FcRH) family in mice demonstrate preferential B cell expression. One of these, FcRH3, encodes a type I transmembrane protein with five extracellular Ig domains and a cytoplasmic tail with a consensus ITIM and a noncanonical ITAM. Analysis of full-length cDNAs from five different mouse strains defines two FcRH3 alleles. A panel of FcRH3-specific mAbs was generated to define its expression pattern and functional potential on B lineage cells. Although poorly detected on the majority of bone marrow or peripheral blood cells, FcRH3 was readily identified on splenic marginal zone (MZ) and MZ precursor B cells, but not on the bulk of newly formed B cells, follicular B cells, germinal center B cells, and plasma cells. In the peritoneal cavity, FcRH3 was found on B1 cells, and not on the majority of B2 cells. Consistent with its possession of an ITIM and ITAM-like sequence, FcRH3 was tyrosine phosphorylated following pervanadate treatment, and its coligation with the BCR inhibited calcium mobilization. These results suggest FcRH3 is a novel immunoregulatory marker of MZ and B1 B lineage cells. PMID- 17082596 TI - Adaptation of solitary intestinal lymphoid tissue in response to microbiota and chemokine receptor CCR7 signaling. AB - Besides Peyer's patches, solitary intestinal lymphoid tissue (SILT) provides a structural platform to efficiently initiate immune responses in the murine small intestine. SILT consists of dynamic lymphoid aggregates that are heterogeneous in size and composition, ranging from small clusters of mostly lineage-negative cells known as cryptopatches to larger isolated lymphoid follicles rich in B cells. In this study, we report that in chemokine receptor CCR7-deficient mice SILT is enlarged, although unchanged in frequency and cellular composition compared with wild-type mice. This phenotype is conferred by bone marrow-derived cells and is independent of the presence of intestinal bacteria. Remarkably, particularly small-sized SILT predominates in germfree wild-type mice. Colonization of wild-type mice with commensal bacteria provokes an adjustment of the spectrum of SILT to that observed under specific pathogen-free conditions by the conversion of pre-existing lymphoid structures into larger-sized SILT. In conclusion, our findings establish that intestinal microbes influence the manifestation of gut-associated lymphoid tissues and identify CCR7 signaling as an endogeneous factor that controls this process. PMID- 17082597 TI - TIRC7 inhibits T cell proliferation by modulation of CTLA-4 expression. AB - Ab targeting of TIRC7 has been shown previously to inhibit T cell proliferation and Th1 lymphocyte-associated cytokine production. In this study, we demonstrate that Ab targeting of TIRC7 induces early cell surface expression of CTLA-4. The majority of stimulated CD4+ and CD8+ human T cells coexpress CTLA-4 and TIRC7. Similar to CTLA-4, TIRC7 rapidly accumulates at the site of Ag adhesion upon T cell activation. TIRC7 seems to colocalize with CTLA-4 in human T cells, and both molecules are associated with clathrin-coated vesicles, indicating they share intracellular transport systems. Moreover, Ab targeting of TIRC7 results in an early activation of CTLA-4 transcription. The inhibition of cell proliferation mediated by TIRC7 is dependent on CTLA-4 expression because the TIRC7-mediated inhibitory effects on cell proliferation and cytokine expression are abolished by Ab blockade of CTLA-4. Splenocytes obtained from CTLA-4-deficient mice are not responsive to TIRC7 Ab targeting. Thus, TIRC7 acts as an upstream regulatory molecule of CTLA-4 expression. PMID- 17082598 TI - Microbial immune suppression mediated by direct engagement of inhibitory Fc receptor. AB - A microbial polysaccharide (glucuronoxylomannan (GXM)) exerts potent immunosuppression by direct engagement to immunoinhibitory receptor FcgammaRIIB. Activation of FcgammaRIIB by GXM leads to the recruitment and phosphorylation of SHIP that prevents IkappaBalpha activation. The FcgammaRIIB blockade inhibits GXM induced IL-10 production and induces TNF-alpha secretion. GXM quenches LPS induced TNF-alpha release via FcgammaRIIB. The addition of mAb to GXM reverses GXM-induced immunosuppression by shifting recognition from FcgammaRIIB to FcgammaRIIA. These findings indicate a novel mechanism by which microbial products can impair immune function through direct stimulation of an inhibitory receptor. Furthermore, our observations provide a new mechanism for the ability of specific Ab to reverse the immune inhibitory effects of certain microbial products. PMID- 17082599 TI - CD8+ IL-17-producing T cells are important in effector functions for the elicitation of contact hypersensitivity responses. AB - Allergen-induced contact hypersensitivity (CHS) is a T cell-mediated delayed-type immune response which has been considered to be primarily mediated by CD8+ T cytotoxic type I (Tc1) cells. IFN-gamma, the prototype Tc1 (Th1) cytokine, has been implicated as the primary inflammatory cytokine for CHS. In this study, we demonstrate that neutralization of IL-17 rather than IFN-gamma suppresses the elicitation of CHS. The suppression does not result from inhibition of the proliferation of allergen-activated T cells. Allergen sensitization induces the development of distinct CD8+ T cell subpopulations that produce IFN-gamma or IL 17. Although CD8+ IL-17-producing cells are stimulated by IL-23, they are inhibited by IL-12, a prototypical stimulator of IFN-gamma-producing Tc1 cells. This indicates that CD8+ IL-17-producing cells are distinct from Tc1 cells and are important in effector functions at the elicitation of CHS. These studies provide insights into a novel mechanism for CHS. PMID- 17082600 TI - Syk is downstream of intercellular adhesion molecule-1 and mediates human rhinovirus activation of p38 MAPK in airway epithelial cells. AB - The airway epithelium is the primary target of inhaled pathogens such as human rhinovirus (HRV). Airway epithelial cells express ICAM-1, the major receptor for HRV. HRV binding to ICAM-1 mediates not only viral entry and replication but also a signaling cascade that leads to enhanced inflammatory mediator production. The specific signaling molecules and pathways activated by HRV-ICAM-1 interactions are not well characterized, although studies in human airway epithelia implicate a role for the p38 MAPK in HRV-induced cytokine production. In the current study, we report that Syk, an important immunoregulatory protein tyrosine kinase, is highly expressed by primary and cultured human airway epithelial cells and is activated in response to infection with HRV16. Biochemical studies revealed that ICAM-1 engagement by HRV and cross-linking Abs enhanced the coassociation of Syk with ICAM-1 and ezrin, a cytoskeletal linker protein. In polarized airway epithelial cells, Syk is diffusely distributed in the cytosol under basal conditions but, following engagement of ICAM-1 by cross-linking Abs, is recruited to the plasma membrane. The enhanced Syk-ICAM-1 association following HRV exposure is accompanied by Syk phosphorylation. ICAM-1 engagement by HRV and cross-linking Abs also induced phosphorylation of p38 in a Syk-dependent manner, and conversely, knockdown of Syk by short interfering (si)RNA substantially diminished p38 activation and IL-8 gene expression. Taken together, these observations identify Syk as an important mediator of the airway epithelial cell inflammatory response by modulating p38 phosphorylation and IL-8 gene expression following ICAM-1 engagement by HRV. PMID- 17082601 TI - A novel model of demyelinating encephalomyelitis induced by monocytes and dendritic cells. AB - Local inflammation may be a precipitating event in autoimmune processes. In this study, we demonstrate that regulated influx of monocytes and dendritic cells (DC) into the CNS causes an acute neurological syndrome that results in a demyelinating encephalomyelitis. Expansion of monocytes and DC by conditional expression of Flt3 ligand in animals expressing CCL2 in the CNS promoted parenchymal cell infiltration and ascending paralysis in 100% of the mice within 9 days of Flt3 ligand induction. Depletion of circulating monocytes and DC reduced disease incidence and severity. Unlike the classical models of experimental autoimmune encephalomyelitis, depletion of CD4+ and CD8+ T cells did not affect disease induction. T cells and demyelinating lesions were observed in the CNS at a later stage as a result of organ-specific inflammation. We propose that alterations in the numbers or function of monocytes and DC coupled to dysregulated expression of chemokines in the neural tissues, favors development of CNS autoimmune disease. PMID- 17082602 TI - TLR4 up-regulation at protein or gene level is pathogenic for lupus-like autoimmune disease. AB - TLR4 is the receptor for the Gram-negative bacterial cell wall component LPS. TLR4 signaling is controlled by both positive and negative regulators to balance optimal immune response and potential sepsis. Unchecked TLR4 activation might result in autoimmune diseases, a hypothesis that has not been formally resolved. In this study, we found that TLR4 signaling to LPS can be positively enforced by expressing gp96 on cell surfaces through the chaperone function of, but not the direct signaling by, gp96; TLR4 as well as the commensal flora are essential for the production of anti-dsDNA Ab and the immune complex-mediated glomerulonephritis in transgenic mice that express surface gp96. Moreover, a similar constellation of autoimmunity was evident in mice that encode multiple copies of tlr4 gene. Our study has revealed that increased TLR4 signaling alone without exogenous insult can break immunological tolerance. It provides a strong experimental evidence for TLR4 dysregulation as an etiology of lupus-like renal disease. PMID- 17082603 TI - Functional interaction of common gamma-chain and growth hormone receptor signaling apparatus. AB - We previously reported on an X-linked SCID (X-SCID) patient, who also had peripheral growth hormone (GH) hyporesponsiveness and abnormalities of the protein phosphorylation events following GH receptor (GHR) stimulation. In the present study, we examined a potential role of common cytokine receptor gamma chain (gammac) in GHR signaling using EBV-transformed lymphocytes from healthy subjects and gammac-negative X-SCID patients. We demonstrated that the proliferative response to GH stimulation of the B cell lines of gammac-negative patients was impaired despite a comparable cellular expression of GHR molecules to controls. In patients, after GH stimulation, no phosphorylation of STAT5 was observed. In addition, the molecule localization through confocal microscopy revealed that in B cell lines of patients no nuclear translocation of STAT5b following GH stimulation occurred differently from controls. Biochemical analysis of the nuclear extracts of gammac-negative cell lines provided further evidence that the amount of STAT5b and its phosphorylated form did not increase following GH stimulation. In patients, cells reconstituted with wild-type gammac abnormal biochemical and functional events were restored resulting in nuclear translocation of STAT5. Confocal experiments revealed that GHR and gammac were colocalized on the cell membrane. Our study demonstrates the existence of a previously unappreciated relationship between GHR-signaling pathway and gammac, which is required for the activation of STAT5b in B cell lines. These data also confirm that growth failure in X-SCID is primarily related to the genetic alteration of the IL2RG gene. PMID- 17082604 TI - Pertussis toxin is superior to TLR ligands in enhancing pathogenic autoimmunity, targeted at a neo-self antigen, by triggering robust expansion of Th1 cells and their cytokine production. AB - Microbial products are assumed to play a major role in triggering pathogenic autoimmunity. Recently accumulated data have shown that these products stimulate the immune system by interacting with TLRs, expressed on APCs. To examine the capacity of various TLR ligands to trigger pathogenic autoimmunity, we used a system in which naive CD4 cells, specific against hen egg lysozyme (HEL), are injected into recipient mice expressing HEL in their eyes. Only when stimulated, the naive cells acquire pathogenic capacity and induce ocular inflammation. Seven TLR ligands were tested in this system: lipoteichoic acid/peptidoglycan, zymosan, poly (I:C), LPS, pertussis toxin (PTX), flagellin, and CpG oligodeoxynucleotide. Treatment of recipient mice with HEL alone stimulated proliferation of the transferred cells, but no disease, whereas ocular inflammation did develop in recipient mice coinjected with HEL and any one of the seven TLR ligands. Inflammation induced by PTX surpassed by its severity those induced by all other tested TLR ligands and was accompanied by a dramatic increase in number of the transferred cells that acquired features of effector Th1 lymphocytes. Ocular inflammation and number of transferred cells in recipients injected with PTX and HEL were substantially reduced by treatment with Abs against IFN-gamma or IL-12, thus indicating the role of these cytokines in the PTX effect. Overall, our observations demonstrate that various TLR ligands are capable of triggering pathogenic autoimmunity and that PTX surpasses other microbial products in this activity, by stimulating excessive proliferation and polarization toward Th1 of naive T cells. PMID- 17082605 TI - Segregation of HLA-C from ICAM-1 at NK cell immune synapses is controlled by its cell surface density. AB - NK cell activity is controlled by the integration of signals from numerous activating and inhibitory receptors at the immunological synapse (IS). However, the importance of segregation and patterning of proteins at the NK cell IS is unknown. In this study, we report that the level of expression of HLA-C on target cells determined its supramolecular organization and segregation from ICAM-1 at the NK cell IS, as well as its capacity to inhibit NK cell cytotoxicity. At YTS NK cell synapses formed with target cells expressing low levels of HLA-C (i.e., 10(4)/cell surface), a multifocal patterning of MHC class I protein predominated, whereas for higher levels of expression (10(5)/cell surface), clusters of HLA-C were more commonly homogeneous, ring-shaped, or containing multiple exclusions. This correlation of protein density with its patterning at the IS was independent of ATP- or actin-driven processes. Importantly, ICAM-1 and HLA-C segregated only at synapses involving target cells expressing high levels of MHC protein. For peripheral blood NK clones, there were specific thresholds in the level of target cell HLA-C needed to inhibit cytotoxicity and to cause segregation of HLA-C from ICAM-1 at the synapse. Thus, the synapse organization of HLA-C, determined by its level of expression, could directly influence NK cell inhibition, e.g., by regulating the proximity of activating and inhibitory receptors. For the first time, this suggests an important function for the assembly of an inhibitory NK cell IS. More broadly, segregation of proteins at intercellular contacts could transmit information about protein expression levels between cells. PMID- 17082606 TI - The study of high-affinity TCRs reveals duality in T cell recognition of antigen: specificity and degeneracy. AB - TCRs exhibit a high degree of Ag specificity, even though their affinity for the peptide/MHC ligand is in the micromolar range. To explore how Ag specificity is achieved, we studied murine T cells expressing high-affinity TCRs engineered by in vitro evolution for binding to hemoglobin peptide/class II complex (Hb/I-Ek). These TCRs were shown previously to maintain Ag specificity, despite having up to 800-fold higher affinity. We compared the response of the high-affinity TCRs and the low-affinity 3.L2 TCR toward a comprehensive set of peptides containing single substitutions at each TCR contact residue. This specificity analysis revealed that the increase in affinity resulted in a dramatic increase in the number of stimulatory peptides. The apparent discrepancy between observed degeneracy in the recognition of single amino acid-substituted Hb peptides and overall Ag specificity of the high-affinity TCRs was examined by generating chimeric peptides between the stimulatory Hb and nonstimulatory moth cytochrome c peptides. These experiments showed that MHC anchor residues significantly affected TCR recognition of peptide. The high-affinity TCRs allowed us to estimate the affinity, in the millimolar range, of immunologically relevant interactions of the TCR with peptide/MHC ligands that were previously unmeasurable because of their weak nature. Thus, through the study of high affinity TCRs, we demonstrated that a TCR is more tolerant of single TCR contact residue substitutions than other peptide changes, revealing that recognition of Ag by T cells can exhibit both specificity and degeneracy. PMID- 17082607 TI - Double-negative T cells, activated by xenoantigen, lyse autologous B and T cells using a perforin/granzyme-dependent, Fas-Fas ligand-independent pathway. AB - The ability to control the response of B cells is of particular interest in xenotransplantation as Ab-mediated hyperacute and acute xenograft rejection are major obstacles in achieving long-term graft survival. Regulatory T cells have been proven to play a very important role in the regulation of immune responses to self or non-self Ags. Previous studies have shown that TCRalphabeta+CD3+CD4 CD8- (double-negative (DN)) T cells possess an immune regulatory function, capable of controlling antidonor T cell responses in allo- and xenotransplantation through Fas-Fas ligand interaction. In this study, we investigated the possibility that xenoreactive DNT cells suppress B cells. We found that DNT cells generated from wild-type C57BL/6 mice expressed B220 and CD25 after rat Ag stimulation. These xenoreactive B220+CD25+ DNT cells lysed activated, but not naive, B and T cells. This killing, which took place through cell-cell contact, required participation of adhesion molecules. Our results indicate that Fas ligand, TGF-beta, TNF-alpha, and TCR-MHC recognition was not involved in DNT cell-mediated syngenic cell killing, but instead this killing was mediated by perforin and granzymes. The xenoreactive DNT cells expressed high levels of granzymes in comparison to allo- or xenoreactive CD8+ T cells. Adoptive transfer of DNT cells in combination with early immune suppression by immunosuppressive analog of 15-deoxyspergualin, LF15-0195, significantly prolonged rat heart graft survival to 62.1 +/- 13.9 days in mice recipients. In conclusion, this study suggests that xenoreactive DNT cells can control B and T cell responses in perforin/granzyme-dependent mechanisms. DNT cells may be valuable in controlling B and T cell responses in xenotransplantation. PMID- 17082608 TI - Stages of germinal center transit are defined by B cell transcription factor coexpression and relative abundance. AB - The transit of T cell-activated B cells through the germinal center (GC) is controlled by sequential activation and repression of key transcription factors, executing the pre- and post-GC B cell program. B cell lymphoma (BCL) 6 and IFN regulatory factor (IRF) 8 are necessary for GC formation and for its molecular activity in Pax5+PU.1+ B cells. IRF4, which is highly expressed in BCL6- GC B cells, is necessary for class switch recombination and the plasma cell differentiation at exit from the GC. In this study, we show at the single-cell level broad coexpression of IRF4 with BCL6, Pax5, IRF8, and PU.1 in pre- and post GC B cells in human and mouse. IRF4 is down-regulated in BCL6+ human GC founder cells (IgD+CD38+), is absent in GC centroblasts, and is re-expressed in positive regulatory domain 1-positive centrocytes, which are negative for all the B cell transcription factors. Activated (CD30+) and activation-induced cytidine deaminase-positive extrafollicular blasts coexpress Pax5 and IRF4. PU.1-negative plasma cells and CD30+ blasts uniquely display the conformational epitope of IRF4 recognized by the MUM1 Ab, an epitope that is absent from any other IRF4+PU.1+ lymphoid and hemopoietic subsets. Low grade B cell lymphomas, representing the malignant counterpart of pre- and post-GC B cells, accordingly express IRF4. However, a fraction of BCL6+ diffuse large B cell lymphomas express IRF4 bearing the MUM1 epitope, indicative of a posttranscriptional modification of IRF4 not seen in the normal counterpart. PMID- 17082609 TI - CCR8 expression identifies CD4 memory T cells enriched for FOXP3+ regulatory and Th2 effector lymphocytes. AB - CD4+ Th2 cells are important regulators of allergic inflammation. CCR8 is thought to play a role in Th2-mediated responses, however, expression of CCR8 in peripheral blood has not been fully characterized. Using a fluorescent form of the ligand selective for CCR8 (F-CCL1), we identified the leukocytes expressing CCR8 in human, monkey, and mouse peripheral blood. CCR8 expression is primarily restricted to a subset of human CD4 memory T lymphocytes (15%). Approximately 40% of CCR8+CD4+ T cells express Th2 cytokines IL-4 or IL-13 while 13% express the Th1 cytokine IFN-gamma. In fact, 50% of all Th2, but only 5% of Th1, cells express CCR8. Upon anti-CD3/anti-CD28 mAb-mediated activation, CCR8+CD4+ T cells secrete 3- to 7-fold higher levels of IL-4, IL-5, IL-9, and IL-13 and 10- to 20 fold lower levels of IFN-gamma or IL-17, compared with CCR8-CD4+ memory T cells. Two-thirds of CCR8+CD4 T cells express cutaneous lymphocyte-associated Ag while the majority lack gut-homing receptors. CCR8+CD4+ cells express CCR7 and CD62L and are present in spleen and lymph nodes of mice. Approximately 25% of CCR8+CD4 T cells express CD25high while 20% of CCR8+CD4+ express the T regulatory cell transcription factor FOXP3 accounting for 60% of all FOXP3-expressing CD4+ T cells. In conclusion, CCR8 marks a diverse subset of CD4 memory T cells enriched for T regulatory and Th2 cells which have the potential for recruitment into sites of allergic inflammation where they could participate in the induction and regulation of the allergic response. PMID- 17082610 TI - Distinct molecular program imposed on CD4+ T cell targets by CD4+CD25+ regulatory T cells. AB - CD4+CD25+ regulatory T cells (Tregs) are key modulators of immunity, but their mechanism of action is unclear. To elucidate the molecular consequences of Treg encounter, we analyzed changes in gene expression in CD4+ T cell targets activated in the presence or absence of CD4+CD25+ Tregs. Tregs did not alter the early activation program of CD4+ T cells, but had reversed many of the activation induced changes by 36 h. It is not known whether Tregs simply induce a set of transcriptional changes common to other nonproliferative states or whether instead Tregs mediate a distinct biological activity. Therefore, we compared the gene profile of T cells following Treg encounter with that of T cells made anergic, TGF-beta-treated, or IL-2-deprived; all possible modes of Treg action. Strikingly, all genes down-regulated in suppressed cells were indeed common to these nonproliferative states. In contrast, Treg encounter led to elevated expression of a unique set of genes in the target T cells. Although different from the nonproliferative states tested, the Treg-imposed gene program is exemplified by expression of many genes associated with growth arrest or inhibition of proliferation. We suggest that Tregs function by the induction of a distinct set of negative regulatory factors that initiate or maintain target T cells in a nonproliferative state. PMID- 17082611 TI - Reversing tumor immune suppression with intratumoral IL-12: activation of tumor associated T effector/memory cells, induction of T suppressor apoptosis, and infiltration of CD8+ T effectors. AB - A single intratumoral injection of IL-12 and GM-CSF-loaded slow-release microspheres induces T cell-dependent eradication of established primary and metastatic tumors in a murine lung tumor model. To determine how the delivery of cytokines directly to the microenvironment of a tumor nodule induces local and systemic antitumor T cell activity, we characterized therapy-induced phenotypic and functional changes in tumor-infiltrating T cell populations. Analysis of pretherapy tumors demonstrated that advanced primary tumors were infiltrated by CD4+ and CD8+ T cells with an effector/memory phenotype and CD4+CD25+Foxp3+ T suppressor cells. Tumor-associated effector memory CD8+ T cells displayed impaired cytotoxic function, whereas CD4+CD25+Foxp3+ cells effectively inhibited T cell proliferation demonstrating functional integrity. IL-12/GM-CSF treatment promoted a rapid up-regulation of CD43 and CD69 on CD8+ effector/memory T cells, augmented their ability to produce IFN-gamma, and restored granzyme B expression. Importantly, treatment also induced a concomitant and progressive loss of T suppressors from the tumor. Further analysis established that activation of pre existing effector memory T cells was short-lived and that both the effector/memory and the suppressor T cells became apoptotic within 4 days of treatment. Apoptotic death of pre-existing effector/memory and suppressor T cells was followed by infiltration of the tumor with activated, nonapoptotic CD8+ effector T lymphocytes on day 7 posttherapy. Both CD8+ T cell activation and T suppressor cell purge were mediated primarily by IL-12 and required IFN-gamma. This study provides important insight into how local IL-12 therapy alters the immunosuppressive tumor milieu to one that is immunologically active, ultimately resulting in tumor regression. PMID- 17082612 TI - Antibodies to complement receptor 3 treat established inflammation in murine models of colitis and a novel model of psoriasiform dermatitis. AB - Prior studies indicated the ability of Abs to complement receptor 3 (CR3, CD11b/CD18) to suppress the production of IL-12 from immune cells. Therefore, we tested the ability of an anti-CR3 Ab (clone M1/70) to treat established IL-12 dependent Th1-mediated inflammation in murine models. Systemic administration of anti-CR3 significantly ameliorated established intestinal inflammation following the intrarectal administration of trinitrobenzene sulfonic acid (TNBS-colitis), as well as colitis and skin inflammation in C57BL/10 RAG-2(-/-) mice reconstituted with CD4+CD45RBhigh T cells. The hyperproliferative skin inflammation in this novel murine model demonstrated many characteristics of human psoriasis, and was prevented by the adoptive transfer of CD45RBlow T cells. In vitro and in vivo studies suggest that anti-CR3 treatment may act, at least in part, by directly inhibiting IL-12 production by APCs. Administration of anti-CR3 may be a useful therapeutic approach to consider for the treatment of inflammatory bowel disease and psoriasis in humans. PMID- 17082613 TI - CD4+CXCR4highCD69+ T cells accumulate in lung adenocarcinoma. AB - The chemokine receptor CXCR4 is involved in the growth and metastasis of tumor cells. However, the expression of its ligand, the chemokine CXCL12, in tumors and its role in regulating the accumulation of immune cells within the tumors is not clear. Using ELISA and immunohistochemistry we found that CXCL12 is expressed in the majority of nonsmall cell lung cancer tissue sections obtained from stage IA to IIB nonsmall cell lung cancer patients undergoing operation. Histopathologic examination of these sections indicated that high CXCL12 expression correlated with increased tumor inflammation. In addition, disease recurrence rates in a subgroup of adenocarcinoma patients showed a tendency to correlate with high CXCL12 expression in the tumor. Isolation of adenocarcinoma-infiltrating immune cells demonstrated an increase in the percentage of CD4+CD69+CXCR4+ T cells as compared with normal lung tissue. About 30% of these cells expressed the regulatory T cell markers CD25high and FoxP3. The percentage of CD8 T cells within the tumor did not change, however; the percentage of NK and NK T cells was significantly reduced. In correlation with CXCR4 expression, CD4 T cells showed increased migration in response to CXCL12 compared with CD8 T cells and NK cells. Overall, these observations suggest that CXCL12 expression may influence tumor progression by shaping the immune cell population infiltrating lung adenocarcinoma tumors. PMID- 17082614 TI - Oligomerization of CXCL10 is necessary for endothelial cell presentation and in vivo activity. AB - The chemokine IFN-gamma-inducible protein of 10 kDa (IP-10; CXCL10) plays an important role in the recruitment of activated T lymphocytes into sites of inflammation by interacting with the G protein-coupled receptor CXCR3. IP-10, like other chemokines, forms oligomers, the role of which has not yet been explored. In this study, we used a monomeric IP-10 mutant to elucidate the functional significance of oligomerization. Although monomeric IP-10 had reduced binding affinity for CXCR3 and heparin, it was able to induce in vitro chemotaxis of activated T cells with the same efficacy as wild-type IP-10. However, monomeric IP-10 was unable to induce recruitment of activated CD8+ T cells into the airways of mice after intratracheal instillation. Use of a different IP-10 mutant demonstrated that this inability was due to lack of oligomerization rather than reduced CXCR3 or heparin binding. Molecular imaging demonstrated that both wild-type and monomeric IP-10 were retained in the lung after intratracheal instillation. However, in vitro binding assays indicated that wild-type, but not monomeric, IP-10 was retained on endothelial cells and could induce transendothelial chemotaxis of activated T cells. We therefore propose that oligomerization of IP-10 is required for presentation on endothelial cells and subsequent transendothelial migration, an essential step for lymphocyte recruitment in vivo. PMID- 17082615 TI - The peptidyl-prolyl isomerase Pin1 regulates granulocyte-macrophage colony stimulating factor mRNA stability in T lymphocytes. AB - Cytokine production is associated with both the normal and pathologic inflammatory response to injury. Previous studies have shown that the immunosuppressants cyclosporin A or FK506, which interact with the peptidyl propyl isomerases cyclophilin A and FK506-binding protein (FKBP12), respectively, block cytokine expression. A third member of the peptidyl-propyl isomerase family, Pin1 is expressed by immune and other cells. Pin1 has been implicated in cell cycle progression, is overexpressed in human tumors, and may rescue neurons from tau-associated degeneration. However, the role of Pin1 in the immune system remains largely unknown. In this study, we analyze the role of Pin1 in GM-CSF expression by human PBMC and CD4+ lymphocytes. We show that Pin1 isomerase activity is necessary for activation-dependent, GM-CSF mRNA stabilization, accumulation, and protein secretion, but not non-AU-rich elements containing cytokine mRNAs, including TGF-beta and IL-4. Mechanistically, Pin1 mediated the association of the AU-rich element-binding protein, AUF1, with GM-CSF mRNA, which determined the rate of decay by the exosome. PMID- 17082616 TI - VH replacement rescues progenitor B cells with two nonproductive VDJ alleles. AB - Inaccurate VDJ rearrangements generate a large number of progenitor (pro)-B cells with two nonproductive IgH alleles. Such cells lack essential survival signals mediated by surface IgM heavy chain (muH chain) expression and are normally eliminated. However, secondary rearrangements of upstream VH gene segments into assembled VDJ exons have been described in mice transgenic for productive muH chains, a process known as VH replacement. If VH replacement was independent of muH chain signals, it could also modify nonproductive VDJ exons and thus rescue pro-B cells with unsuccessful rearrangements on both alleles. To test this hypothesis, we homologously replaced the JH cluster of a mouse with a nonproductive VDJ exon. Surprisingly, B cell development in IgHVDJ-/VDJ- mice was only slightly impaired and significant numbers of IgM-positive B cells were produced. DNA sequencing confirmed that all VDJ sequences from muH chain-positive B lymphoid cells were generated by VH replacement in a RAG-dependent manner. Another unique feature of our transgenic mice was the presence of IgH chains with unusually long CDR3-H regions. Such IgH chains were functional and only modestly counter-selected, arguing against a strict length constraint for CDR3-H regions. In conclusion, VH replacement can occur in the absence of a muH chain signal and provides a potential rescue mechanism for pro-B cells with two nonproductive IgH alleles. PMID- 17082617 TI - HLA-B*2704, an allotype associated with ankylosing spondylitis, is critically dependent on transporter associated with antigen processing and relatively independent of tapasin and immunoproteasome for maturation, surface expression, and T cell recognition: relationship to B*2705 and B*2706. AB - B*2704 is strongly associated to ankylosing spondylitis in Asian populations. It differs from the main HLA-B27 allotype, B*2705, in three amino acid changes. We analyzed the influence of tapasin, TAP, and immunoproteasome induction on maturation, surface expression, and T cell allorecognition of B*2704 and compared some of these features with B*2705 and B*2706, allotypes not associated to disease. In the tapasin-deficient .220 cell line, this chaperone significantly influenced the extent of folding of B*2704 and B*2705, but not their egress from the endoplasmic reticulum. In contrast, B*2706 showed faster folding and no accumulation in the endoplasmic reticulum in the absence of tapasin. Surface expression of B*2704 was more tapasin dependent than B*2705. However, expression of free H chain decreased in the presence of this chaperone for B*2705 but not B*2704, suggesting that more suboptimal ligands were loaded on B*2705 in the absence of tapasin. Despite its influence on surface expression, tapasin had little effect on allorecognition of B*2704. Both surface expression and T cell recognition of B*2704 were critically dependent on TAP, as established with TAP deficient and TAP-proficient T2 cells. Both immunoproteasome and surface levels of B*2704 were induced by IFN-gamma, but this had little effect on allorecognition. Thus, except for the differential effects of tapasin on surface expression, the tapasin, TAP, and immunoproteasome dependency of B*2704 for maturation, surface expression, and T cell recognition are similar to B*2705, indicating that basic immunological features are shared by the two major HLA-B27 allotypes associated to ankylosing spondylitis in human populations. PMID- 17082618 TI - Activating transcription factor/cAMP response element binding protein family member regulated transcription of CD1A. AB - CD1a has a unique expression pattern among Ag-presenting molecules, expressed specifically on cortical thymocytes and APCs. As autoimmune disease, infection, and tumors can all result in alteration of CD1a expression, we are attempting to characterize the transcriptional regulation, and thus shed some light on specific expression, of CD1A. In this study, we have identified a minimal proximal promoter region required for CD1A transcription. Computer searches within this region identified numerous potential binding sites for lymphoid-specific transcription factors, including the ETS transcription factors, C/EBP, GATA, and CREB. Deletion and site-specific mutant analysis revealed a critical role of a potential cAMP response element (CRE) 965 bp upstream of the CD1A translation start site. Two activating transcription factor (ATF)/CREB family members, CREB-1 and ATF-2, are able to bind this site in vitro and in vivo. Notably, activation of ATF/CREB family members decreases CD1A transcription, while decrease in ATF-2 expression results in increased CD1A RNA level. The fact that these factors also bind the CD1A promoter in human monocytes strongly suggests a role for ATF/CREB family members in regulation of CD1A expression. PMID- 17082619 TI - Genes within the Idd5 and Idd9/11 diabetes susceptibility loci affect the pathogenic activity of B cells in nonobese diabetic mice. AB - Autoreactive T cells clearly mediate the pancreatic beta cell destruction causing type 1 diabetes (T1D). However, studies in NOD mice indicate that B cells also contribute to pathogenesis because their ablation by introduction of an Igmunull mutation elicits T1D resistance. T1D susceptibility is restored in NOD.Igmunull mice that are irradiated and reconstituted with syngeneic bone marrow plus NOD B cells, but not syngeneic bone marrow alone. Thus, we hypothesized some non-MHC T1D susceptibility (Idd) genes contribute to disease by allowing development of pathogenic B cells. Supporting this hypothesis was the finding that unlike those from NOD donors, engraftment with B cells from H2g7 MHC-matched, but T1D resistant, nonobese-resistant (NOR) mice failed to restore full disease susceptibility in NOD.Igmunull recipients. T1D resistance in NOR mice is mainly encoded within the Idd13, Idd5.2, and Idd9/11 loci. B cells from NOD congenic stocks containing Idd9/11 or Idd5.1/5.2-resistance loci, respectively, derived from the NOR or C57BL/10 strains were characterized by suppressed diabetogenic activity. Immature autoreactive B cells in NOD mice have an impaired ability to be rendered anergic upon Ag engagement. Interestingly, both Idd5.1/5.2 and Idd9/11-resistance loci were found to normalize this B cell tolerogenic process, which may represent a mechanism contributing to the inhibition of T1D. PMID- 17082620 TI - Fine mapping of collagen-induced arthritis quantitative trait loci in an advanced intercross line. AB - The generation of advanced intercross lines (AIL) is a powerful approach for high resolution fine mapping of quantitative trait loci (QTLs), because they accumulate much more recombination events compared with conventional F2 intercross and N2 backcross. However, the application of this approach is severely hampered by the requirements of excessive resources to maintain such crosses, i.e., in terms of animal care, space, and time. Therefore, in this study, we produced an AIL to fine map collagen-induced arthritis (CIA) QTLs using comparatively limited resources. We used only 308 (DBA/1 x FVB/N)F11/12 AIL mice to refine QTLs controlling the severity and onset of arthritis as well as the Ab response and T cell subset in CIA, namely Cia2, Cia27, and Trmq3. These QTLs were originally identified in (DBA/1 x FVB/N)F2 progeny. The confidence intervals of the three QTLs were refined from 40, 43, and 48 Mb to 12, 4.1, and 12 Mb, respectively. The data were complemented by the use of another QTL fine-mapping approach, haplotype analysis, to further refine Cia2 into a 2-Mb genomic region. To aid in the search for candidate genes for the QTLs, genome-wide expression profiling was performed to identify strain-specific differentially expressed genes within the confidence intervals. Of the 1396 strain-specific differentially expressed genes, 3, 3, and 12 genes were within the support intervals of the Cia2, Cia27, and Trmq3, respectively. In addition, this study revealed that Cia27 and Trmq3 controlling anti-CII IgG2a Ab and CD4:CD8 T cell ratio, respectively, also regulated CIA clinical phenotypes. PMID- 17082621 TI - The immunosuppressant cyclosporin A antagonizes human formyl peptide receptor through inhibition of cognate ligand binding. AB - Cyclosporin A (CsA) is a fungus-derived cyclic undecapeptide with potent immunosuppressive activity. Its analog, cyclosporin H (CsH), lacks immunosuppressive function but can act as an antagonist for the human formyl peptide receptor (FPR). More recent studies have shown that CsA also inhibits fMLF-induced degranulation in differentiated HL-60 promyelocytic leukemia cells. However, it is unclear whether CsA interferes with ligand-receptor interaction, G protein activation, or other downstream signaling events. In this study we used human neutrophils, differentiated HL-60 cells, and rat basophilic leukemia (RBL) 2H3 cells expressing human FPR (RBL-FPR) to identify the action site of CsA. In functional assays, CsA inhibited fMLF-stimulated degranulation, chemotaxis, calcium mobilization, and phosphorylation of the MAPKs ERK 1/2 and the serine/threonine protein kinase Akt. CsA also blocked Trp-Lys-Tyr-Met-Val-D-Met (WKYMVm)-induced functions in RBL-FPR cells. Concentrations for half-maximal inhibition with CsA are generally 6- to 50-fold higher than that of CsH. CsA was compared with another immunosuppressant, ascomycin, relative to the inhibitory effects on FPR-mediated chemotaxis, calcium mobilization, and degranulation. In these experiments, ascomycin produced no inhibitory effects at low micromolar concentrations (1-4 microM), whereas the inhibitory effects of CsA were prominent at comparable concentrations. Finally, CsA dose-dependently inhibited the uptake of fNle-Leu-Phe-Nle-Tyr-Lys-fluoresceine and [3H]fMLF or [125I]WKYMVm binding to FPR. However, CsA and CsH did not show any obvious inhibitory effect on FPR-like 1-mediated cellular functions. These results demonstrate that CsA is a selective antagonist of FPR and that its inhibition of fMLF-stimulated leukocyte activation is at the level of cognate ligand binding. PMID- 17082622 TI - TLR-dependent induction of IFN-beta mediates host defense against Trypanosoma cruzi. AB - Host resistance to the intracellular protozoan parasite Trypanosoma cruzi depends on IFN-gamma production by T cells and NK cells. However, the involvement of innate immunity in host resistance to T. cruzi remains unclear. In the present study, we investigated host defense against T. cruzi by focusing on innate immunity. Macrophages and dendritic cells (DCs) from MyD88(-/-)TRIF(-/-) mice, in which TLR-dependent activation of innate immunity was abolished, were defective in the clearance of T. cruzi and showed impaired induction of IFN-beta during T. cruzi infection. Neutralization of IFN-beta in MyD88(-/-) macrophages led to enhanced T. cruzi growth. Cells from MyD88(-/-)IFNAR1(-/-) mice also showed impaired T. cruzi clearance. Furthermore, both MyD88(-/-)TRIF(-/-) and MyD88(-/ )IFNAR1(-/-) mice were highly susceptible to in vivo T. cruzi infection, highlighting the involvement of innate immune responses in T. cruzi infection. We further analyzed the molecular mechanisms for the IFN-beta-mediated antitrypanosomal innate immune responses. MyD88(-/-)TRIF(-/-) and MyD88(-/ )IFNAR1(-/-) macrophages and DCs exhibited defective induction of the GTPase IFN inducible p47 (IRG47) after T. cruzi infection. RNA interference-mediated reduction of IRG47 expression in MyD88(-/-) macrophages resulted in increased intracellular growth of T. cruzi. These findings suggest that TLR-dependent expression of IFN-beta is involved in resistance to T. cruzi infection through the induction of IRG47. PMID- 17082623 TI - Adoptive transfer of CD8alpha+ dendritic cells (DC) isolated from mice infected with Chlamydia muridarum are more potent in inducing protective immunity than CD8alpha- DC. AB - Chlamydial infections are serious public health concerns worldwide. In this study, we examined the role of dendritic cell (DC) subsets in inducing protective immunity against chlamydial infection using an adoptive transfer approach. We found that CD11c+CD8alpha+ (double-positive, DP) DC, compared with CD11c+CD8alpha (single-positive, SP) DC isolated from infected mice, are more potent inducers of protective immunity. Specifically, mice pretreated with DPDC from infected mice, upon infection with Chlamydia trachomatis mouse pneumonitis (MoPn), experienced significantly less severe body weight loss and in vivo chlamydial growth. Analysis of MoPn-driven cytokine production by immune cells revealed that mice that were treated with DPDC produced significantly higher levels of Th1 (TNF alpha, IFN-gamma, and IL-12) but lower levels of Th2 (IL-4, IL-5, and IL-13) related cytokines than the recipients of SPDC following infection challenge. Moreover, DPDC-treated mice displayed significantly higher levels of MoPn specific IgG2a production and delayed-type hypersensitivity responses compared with SPDC-treated mice. Furthermore, DPDC isolated from infected mice produced higher amounts of IL-12 and IL-10 in vitro in comparison with SPDC. These data indicate that CD8alpha+ DC have a significantly higher capacity in inducing protective immunity compared with CD8alpha- DC, demonstrating the crucial role of DC1-like cells in eliciting protection against C. trachomatis infection. PMID- 17082624 TI - IL-10 deficiency promotes increased Borrelia burgdorferi clearance predominantly through enhanced innate immune responses. AB - Borrelia burgdorferi is capable of persistently infecting a variety of hosts despite eliciting potent innate and adaptive immune responses. Preliminary studies indicated that IL-10-deficient (IL-10(-/-)) mice exhibit up to 10-fold greater clearance of B. burgdorferi from target tissues compared with wild-type mice, establishing IL-10 as the only cytokine currently known to have such a significant effect on spirochetal clearance. To further delineate these IL-10 mediated immune effects, kinetic studies indicated that spirochete dissemination to target tissues is similar in both wild-type and IL-10(-/-) mouse strains, and that enhanced clearance of B. burgdorferi in IL-10(-/-) mice is correlated with increased B. burgdorferi-specific Ab as early as 2 wk postinfection. Immunoblot analysis indicated that Abs produced by infected IL-10(-/-) and wild-type mice recognize similar ranges of spirochetal Ags. Immune sera from IL-10(-/-) and wild type mice also exhibited similar bactericidal activity in vitro, and passive transfer of these immune sera into B. burgdorferi-infected SCID mice caused similar reductions of bacterial numbers in target tissues. Infectious dose studies indicated that 8-fold more B. burgdorferi were needed to efficiently infect naive IL-10(-/-) mice, suggesting these animals possess higher innate barriers to infection. Moreover, macrophages derived from IL-10(-/-) mice exhibit enhanced proinflammatory responses to B. burgdorferi stimulation compared with wild-type controls, and these responses are not significantly affected by the presence of immune serum. These findings confirm that B. burgdorferi clearance by innate immune responses is more efficient in the absence of IL-10, and these activities are not directly related to increased levels of B. burgdorferi specific Ab. PMID- 17082625 TI - NK cell-derived IFN-gamma differentially regulates innate resistance and neutrophil response in T cell-deficient hosts infected with Mycobacterium tuberculosis. AB - Although it is known that IFN-gamma-secreting T cells are critical for control of Mycobacterium tuberculosis infection, the contribution of IFN-gamma produced by NK cells to host resistance to the pathogen is less well understood. By using T cell-deficient RAG(-/-) mice, we showed that M. tuberculosis stimulates NK cell dependent IFN-gamma production in naive splenic cultures and in lungs of infected animals. More importantly, common cytokine receptor gamma-chain(-/-)RAG(-/-) animals deficient in NK cells, p40(-/-)RAG(-/-), or anti-IFN-gamma mAb-treated RAG(-/-) mice displayed significantly increased susceptibility to M. tuberculosis infection compared with untreated NK-sufficient RAG(-/-) controls. Studies comparing IL-12 p40- and p35-deficient RAG(-/-) mice indicated that IL-12 plays a more critical role in the induction of IFN-gamma-mediated antimycobacterial effector functions than IL-23 or other p40-containing IL-12 family members. The increased susceptibility of IL-12-deficient or anti-IFN-gamma mAb-treated RAG(-/ ) mice was associated not only with elevated bacterial loads, but also with the development of granulocyte-enriched foci in lungs. This tissue response correlated with increased expression of the granulocyte chemotactic chemokines KC and MIP-2 in NK as well as other leukocyte populations. Interestingly, depletion of granulocytes further increased bacterial burdens and exacerbated pulmonary pathology in these animals, revealing a compensatory function for neutrophils in the absence of IFN-gamma. The above observations indicate that NK cell-derived IFN-gamma differentially regulates T-independent resistance and granulocyte function in M. tuberculosis infection and suggest that this response could serve as an important barrier in AIDS patients or other individuals with compromised CD4+ T cell function. PMID- 17082626 TI - Human cytomegalovirus envelope glycoproteins B and H are necessary for TLR2 activation in permissive cells. AB - Human CMV (HCMV) is a ubiquitous member of the Herpesviridae family and an opportunistic pathogen that poses significant health risks for immunocompromised patients. HCMV pathogenesis is intimately tied to the immune status of the host, thus characterization of the innate immune response to HCMV infection is critical for understanding disease progression. Previously, we identified TLR2 as a host factor that detects and initiates inflammatory cytokine secretion in response to HCMV independent of viral replication. In this study, we show that two entry mediating envelope gp, gp B (gB) and gp H (gH), display determinants recognized by TLR2. Neutralizing Abs against TLR2, gB and gH inhibit inflammatory cytokine responses to HCMV infection, suggesting that inflammatory cytokine stimulation by HCMV is mediated by interactions between these envelope gp and TLR2. Furthermore, both gB and gH coimmunoprecipitate with TLR2 and TLR1, indicating that these envelope gp directly interact with TLR2 and that a TLR2/TLR1 heterodimer is a functional sensor for HCMV. Because our previous studies were conducted in model cell lines, we also show that TLR2 is expressed by HCMV permissive human fibroblast cell strains, and that TLR2 is a functional sensor in these cells. This study further elucidates the importance and potency of envelope gp as a class of molecules displaying pathogen-associated molecular patterns that are recognized with immediate kinetics by TLRs in permissive cells. PMID- 17082627 TI - HIV induces maturation of monocyte-derived dendritic cells and Langerhans cells. AB - In HIV infection, dendritic cells (DCs) may play multiple roles, probably including initial HIV uptake in the anogenital mucosa, transport to lymph nodes, and subsequent transfer to T cells. The effects of HIV-1 on DC maturation are controversial, with several recent conflicting reports in the literature. In this study, microarray studies, confirmed by real-time PCR, demonstrated that the genes encoding DC surface maturation markers were among the most differentially expressed in monocyte-derived dendritic cells (MDDCs), derived from human blood, treated with live or aldrithriol-2-inactivated HIV-1(BaL). These effects translated to enhanced cell surface expression of these proteins but differential expression of maturation markers was only partial compared with the effects of a conventional potent maturation stimulus. Such partially mature MDDCs can be converted to fully mature cells by this same potent stimulus. Furthermore, live HIV-1 stimulated greater changes in maturation marker surface expression than aldrithriol-2-inactivated HIV-1 and this enhanced stimulation by live HIV-1 was mediated via CCR5, thus suggesting both viral replication-dependent and independent mechanisms. These partially mature MDDCs demonstrated enhanced CCR7 mediated migration and are also able to stimulate interacting T cells in a MLR, suggesting DCs harboring HIV-1 might prepare CD4 lymphocytes for transfer of HIV 1. Increased maturation marker surface expression was also demonstrated in native DCs, ex vivo Langerhans cells derived from human skin. Thus, HIV initiates maturation of DCs which could facilitate subsequent enhanced transfer to T cells. PMID- 17082628 TI - Flavivirus activation of plasmacytoid dendritic cells delineates key elements of TLR7 signaling beyond endosomal recognition. AB - TLR7 senses RNA in endosomal compartments. TLR7 expression and signaling have been demonstrated in plasmacytoid and myeloid dendritic cells, B cells, and T cells. The regulation of TLR7 signaling can play a crucial role in shaping the immune response to RNA viruses with different cellular tropisms, and in developing adjuvants capable of promoting balanced humoral and cell-mediated immunity. We used unique characteristics of two ssRNA viruses, dengue virus and influenza virus, to delineate factors that regulate viral RNA-human TLR7 signaling beyond recognition in endosomal compartments. Our data show that TLR7 recognition of enveloped RNA virus genomes is linked to virus fusion or uncoating from the endosome. The signaling threshold required to activate TLR7-type I IFN production is greater than that required to activate TLR7-NF-kappaB-IL-8 production. The higher order structure of viral RNA appears to be an important determinant of TLR7-signaling potency. A greater understanding of viral RNA-TLR7 activity relationships will promote rational approaches to interventional and vaccine strategies for important human viral pathogens. PMID- 17082629 TI - Sertoli cells initiate testicular innate immune responses through TLR activation. AB - TLRs play a crucial role in early host defense against invading pathogens. In the seminiferous epithelium, Sertoli cells are the somatic nurse cells that mechanically segregate germ cell autoantigens by means of the blood-tubular barrier and create a microenvironment that protects germ cells from both interstitial and ascending invading pathogens. The objective of this study was to examine TLR expression and their functional responses to specific agonists in mouse Sertoli cells. We measured the expression of TLR2, TLR4, TLR5, and TLR6 mRNAs and confirmed by FACS analysis the presence of proteins TLR2 and TLR5 on which we focused our study. Stimulation of Sertoli cells with macrophage activating lipopeptide-2, agonist of TLR2/TLR6, and with flagellin, agonist of TLR5, induces augmented secretion of the chemokine MCP-1. To assess the functional significance of MCP-1 production following TLR stimulation, conditioned medium from either macrophage-activating lipopeptide-2 or flagellin treated Sertoli cells was tested for in vitro chemotaxis assay, and a significant increase of macrophage migration was observed in comparison with unstimulated conditioned medium. Moreover, we studied the role of NF-kappaB and of MAPKs in regulating TLR-mediated MCP-1 secretion by using inhibitors specific for each transduction pathway and we demonstrated a pivotal role of the IkappaB/NF-kappaB and JNK systems. In addition, TLR2/TLR6 and TLR5 stimulation induces increased ICAM-1 expression in Sertoli cells. Collectively, this study demonstrates the novel ability of Sertoli cells to potentially respond to a wide variety of bacteria through TLR stimulation. PMID- 17082630 TI - Bordetella bronchiseptica modulates macrophage phenotype leading to the inhibition of CD4+ T cell proliferation and the initiation of a Th17 immune response. AB - Bordetella bronchiseptica is a Gram-negative bacterium equipped with several colonization factors that allow it to establish a persistent infection of the murine respiratory tract. Previous studies indicate that B. bronchiseptica adenylate cyclase toxin (ACT) and the type III secretion system (TTSS) synergize to drive dendritic cells into an altered phenotype to down-regulate the host immune response. In this study, we examined the effects of B. bronchiseptica ACT and TTSS on murine bone marrow-derived macrophages. We demonstrate that ACT and TTSS are required for the inhibition of Ag-driven CD4+ T cell proliferation by bacteria-infected macrophages. We identify PGE2 as the mediator of this inhibition, and we show that ACT and the TTSS synergize to increase macrophage production of PGE2. We further demonstrate that B. bronchiseptica can modulate normal macrophage function and drive the immune response toward a Th17 phenotype classified by the significant production of IL-17. In this study, we show that B. bronchiseptica-infected macrophages can induce IL-17 production from naive CD4+ splenocytes, and that lung tissues from B. bronchiseptica-infected mice exhibit a strong Th17 immune response. ACT inhibited surface expression of CD40 and CD86, suppressed TNF-alpha production, and up-regulated IL-6 production. TTSS also synergized with ACT to up-regulate IL-10 and PGE2 secretion. These findings indicate that persistent colonization by B. bronchiseptica may rely on the ability of the bacteria to differentially modulate both macrophage and dendritic cell function leading to an altered adaptive immune response and subsequent bacterial colonization. PMID- 17082631 TI - Prenatal malaria immune experience affects acquisition of Plasmodium falciparum merozoite surface protein-1 invasion inhibitory antibodies during infancy. AB - African infants are often born of mothers infected with malaria during pregnancy. This can result in fetal exposure to malaria-infected erythrocytes or their soluble products with subsequent fetal immune priming or tolerance in utero. We performed a cohort study of 30 newborns from a malaria holoendemic area of Kenya to determine whether T cell sensitization to Plasmodium falciparum merozoite surface protein-1 (MSP-1) at birth correlates with infant development of anti-MSP 1 Abs acquired as a consequence of natural malaria infection. Abs to the 42- and 19-kDa C-terminal processed fragments of MSP-1 were determined by serology and by a functional assay that quantifies invasion inhibition Abs against the MSP-1(19) merozoite ligand (MSP-1(19) IIA). Infants had detectable IgG and IgM Abs to MSP 1(42) and MSP-1(19) at 6 mo of age with no significant change by age 24-30 mo. In contrast, MSP-1(19) IIA levels increased from 6 to 24-30 mo of age (16-29%, p < 0.01). Infants with evidence of prenatal exposure to malaria (defined by P. falciparum detection in maternal, placental, and/or cord blood compartments) and T cell sensitization at birth (defined by cord blood lymphocyte cytokine responses to MSP-1) showed the greatest age-related increase in MSP-1(19) IIA compared with infants with prenatal exposure to malaria but who lacked detectable T cell MSP-1 sensitization. These data suggest that fetal sensitization or tolerance to MSP-1, associated with maternal malaria infection during pregnancy, affects the development of functional Ab responses to MSP-1 during infancy. PMID- 17082632 TI - Cytosolic localization of Listeria monocytogenes triggers an early IFN-gamma response by CD8+ T cells that correlates with innate resistance to infection. AB - IFN-gamma is critical for innate immunity against Listeria monocytogenes (L. monocytogenes), and it has long been thought that NK cells are the major source of IFN-gamma during the first few days of infection. However, it was recently shown that a significant number of CD44highCD8+ T cells also secrete IFN-gamma in an Ag-independent fashion within 16 h of infection with L. monocytogenes. In this report, we showed that infection with other intracellular pathogens did not trigger this early IFN-gamma response and that cytosolic localization of Listeria was required to induce rapid IFN-gamma production by CD44highCD8+ T cells. Infection of C57BL/6 mice with an Escherichia coli strain expressing listeriolysin O (LLO), a pore-forming toxin from L. monocytogenes, also resulted in rapid IFN-gamma expression by CD8+ T cells. These results suggest that LLO expression is essential for induction of the early IFN-gamma response, although it is not yet clear whether LLO plays a direct role in triggering a signal cascade that leads to cytokine production or whether it is required simply to release other bacterial product(s) into the host cell cytosol. Interestingly, mouse strains that displayed a rapid CD8+ T cell IFN-gamma response (C57BL/6, 129, and NZB) all had lower bacterial burdens in the liver 3 days postinfection compared with mouse strains that did not have an early CD8+ T cell IFN-gamma response (BALB/c, A/J, and SJL). These data suggest that participation of memory CD8+ T cells in the early immune response against L. monocytogenes correlates with innate host resistance to infection. PMID- 17082633 TI - Lipopolysaccharide-activated CD4+CD25+ T regulatory cells inhibit neutrophil function and promote their apoptosis and death. AB - CD4+CD25+ T regulatory (Treg) cells play a central role in the suppression of immune response and prevention of autoimmune reactions. Pathogen recognition receptors expressed by immune cells, such as TLRs, may provide a critical link between the innate and adaptive immune systems. There is also evidence that TLR ligands can directly modulate the suppressive capacity of Treg cells. Here, we showed that CD4+CD25+ Treg cells affect neutrophil function and survival and that the TLR4 ligand is involved in the regulation of the cell interactions. We found that LPS-activated Treg cells inhibit reactive oxygen intermediates and cytokine production by neutrophils. Moreover, Treg cells reverse LPS-induced survival of neutrophils and promote their apoptosis and death. We also found that TCR activated Treg cells induce the same effects on polymorphonuclear neutrophils as those achieved by TLR4 stimulation. Importantly, the suppressive potential of CD4+CD25+ Treg cells induced by LPS seems to be partially IL-10 and TGF-beta dependent, whereas anti-CD3/CD28 stimulation is rather contact dependent. Together, these observations suggest that Treg cells have the ability to directly regulate neutrophil function and life span when both types of the cells are exposed to LPS. PMID- 17082634 TI - Airway epithelial cells produce B cell-activating factor of TNF family by an IFN beta-dependent mechanism. AB - Activation of B cells in the airways is now believed to be of great importance in immunity to pathogens, and it participates in the pathogenesis of airway diseases. However, little is known about the mechanisms of local activation of B cells in airway mucosa. We investigated the expression of members of the B cell activating TNF superfamily (B cell-activating factor of TNF family (BAFF) and a proliferation-inducing ligand (APRIL)) in resting and TLR ligand-treated BEAS-2B cells and primary human bronchial epithelial cells (PBEC). In unstimulated cells, expression of BAFF and APRIL was minimal. However, BAFF mRNA was significantly up regulated by TLR3 ligand (dsRNA), but not by other TLR ligands, in both BEAS-2B cells (376-fold) and PBEC (224-fold). APRIL mRNA was up-regulated by dsRNA in PBEC (7-fold), but not in BEAS-2B cells. Membrane-bound BAFF protein was detectable after stimulation with dsRNA. Soluble BAFF protein was also induced by dsRNA (> 200 pg/ml). The biological activity of the epithelial cell-produced BAFF was verified using a B cell survival assay. BAFF was also strongly induced by IFN beta, a cytokine induced by dsRNA. Induction of BAFF by dsRNA was dependent upon protein synthesis and IFN-alphabeta receptor-JAK-STAT signaling, as indicated by studies with cycloheximide, the JAK inhibitor I, and small interfering RNA against STAT1 and IFN-alphabeta receptor 2. These results suggest that BAFF is induced by dsRNA in airway epithelial cells and that the response results via an autocrine pathway involving IFN-beta. The production of BAFF and APRIL by epithelial cells may contribute to local accumulation, activation, class switch recombination, and Ig synthesis by B cells in the airways. PMID- 17082635 TI - IL-1 beta and TNF-alpha regulation of the adenosine receptor (A2A) expression: differential requirement for NF-kappa B binding to the proximal promoter. AB - Adenosine is a potent endogenous regulator of airway inflammation that acts through specific receptor subtypes that can either cause constriction (A1R, A2BR, and A3R) or relaxation (A2AR) of the airways. We therefore examined the effects of key inflammatory mediators on the expression of the A2AR in a lung epithelial cell line (A549). IL-1beta and TNF-alpha increased the expression of the A2AR gene at the mRNA and protein levels. In contrast, LPS had no effect on A2AR gene expression. IL-1beta and TNF-alpha rapidly activated p50 and p65, but not C-Rel, RelB, or p52, and both IL-1beta- and TNF-alpha-stimulated A2AR expression was inhibited by the IkappaB kinase 2 inhibitor AS602868 in a concentration-dependent manner. Using chromatin immunoprecipitation assays, we demonstrate that IL-1beta can enhance p65 association with putative kappaB binding sites in the A2AR promoter in a temporal manner. In contrast, TNF-alpha failed to enhance p65 binding to these putative sites. Functionally, the two most 5' kappaB sites were important for IL-1beta-, but not TNF-alpha-, induced A2AR promoter reporter gene activity. Finally, neither TNF-alpha nor Il-1beta had any effect on A2AR mRNA transcript degradation. These results directly implicate a major role for NF kappaB in the regulation of A2AR gene transcription by IL-1beta and TNF-alpha but suggest that the effects of TNF-alpha on A2AR gene transcription are not mediated through the proximal promoter. PMID- 17082636 TI - Extracellular heat shock protein-70 induces endotoxin tolerance in THP-1 cells. AB - Recent data suggest that heat shock protein-70 (HSP-70), an intracellular protein, can exist in the extracellular compartment and signal through the CD14/TLR4 pathway. In this study, we tested the hypothesis that extracellular HSP 70 induces endotoxin (LPS) tolerance. Using human monocyte cell line (THP-1), initial dose-response experiments were conducted to determine a subthreshold concentration of HSP-70 that does not induce NF-kappaB activity. Differentiated THP-1 cells were preconditioned with subthreshold concentration (0.03 microg/ml HSP-70) for 18 h, followed by LPS stimulation (1 microg/ml) for 4 h. Preconditioning with HSP-70 decreased subsequent LPS-mediated NF-kappaB-dependent promoter activity and was accompanied by significant decreases of supernatant TNF levels. Furthermore, human monocytes isolated from human volunteers, subsequently preconditioned with HSP-70, demonstrated LPS tolerance as evidenced by abrogated supernatant TNF levels. Additional experiments were conducted to exclude the possibility of endotoxin contamination of HSP-70 by boiling HSP-70 at 100 degrees C for 1 h or preconditioning with equivalent concentrations of endotoxin as present in the HSP-70 preparation. These experiments indicated that induction of tolerance was not secondary to endotoxin contamination. Neutralization experiments with an anti-HSP-70 Ab confirmed the specificity of HSP-70 in tolerance induction. Preconditioning with HSP-70 attenuated cytosolic degradation of inhibitor kappaB-alpha and inhibited activation of inhibitor kappaB kinase following LPS stimulation. HSP-70 preconditioning decreased phosphorylation of the p65 subunit of NF-kappaB following LPS stimulation. These data suggest a novel role for extracellular HSP-70 in modifying mononuclear cell responses to subsequent LPS challenge. PMID- 17082637 TI - A JNK-independent signaling pathway regulates TNF alpha-stimulated, c-Jun-driven FRA-1 protooncogene transcription in pulmonary epithelial cells. AB - Among the several effectors that mediate TNF-alpha action is AP-1, which consists of transcription factors belonging to the JUN and FOS families. Although the effects of TNF-alpha in immune cells, such as the induction of NF-kappaBeta, are well known, the mechanisms by which it induces transcriptional activation of AP-1 in pulmonary epithelial cells are not well defined. In this study, we report that TNF-alpha stimulates the expression of the FRA-1 protooncogene in human pulmonary epithelial cells using c-Jun, acting via a 12-O-tetradecanoylphorbol-13 acetate response element located at -318. Although TNF-alpha stimulates phosphorylation of c-Jun, the inhibition of JNK activity had no significant effect on FRA-1 induction. Consistent with this result, ectopic expression of a c-Jun mutant lacking JNK phosphorylation sites had no effect on the TNF-alpha-induced expression of the promoter. In contrast, inhibition of the ERK pathway or ectopic expression of an ERK1 mutant strikingly reduced FRA-1 transcription. ERK inhibition not only blocked phosphorylation of Elk1, CREB, and ATF1, which constitutively bind to the FRA-1 promoter, but also suppressed the recruitment of c-Jun to the promoter. We found that short interfering RNA-mediated silencing of FRA-1 enhances TNF-alpha-induced IL-8 expression, whereas overexpression causes an opposite effect. Our findings collectively indicate that ERK signaling plays key roles in both Elk1, CREB, and ATF-1 activation and the subsequent recruitment of c-Jun to the FRA-1 promoter in response to TNF-alpha in pulmonary epithelial cells. PMID- 17082638 TI - Bruton's tyrosine kinase is required for TLR-induced IL-10 production. AB - Bruton's tyrosine kinase (Btk) is a critical signaling mediator downstream of the B cell Ag receptor. X-linked agammaglobulinemia is caused by mutations in Btk resulting in multiple defects in B cell development and function, and recurrent bacterial infections. Recent evidence has also supported a role for Btk in TLR signaling. We demonstrate that Btk is activated by TLR4 in primary macrophages and is required for normal TLR-induced IL-10 production in multiple macrophage populations. Btk-deficient bone marrow-derived macrophages secrete decreased levels of IL-10 in response to multiple TLR ligands, compared with wild-type (WT) cells. Similarly, Btk-deficient peritoneal and splenic macrophages secrete decreased IL-10 levels compared with WT cultures. This phenotype correlates with Btk-dependent induction of NF-kappaB and AP-1 DNA binding activity, and altered commensal bacteria populations. Decreased IL-10 production may be responsible for increased IL-6 because blocking IL-10 in WT cultures increased IL-6 production, and supplementation of IL-10 to Btk-deficient cultures decreased IL-6 production. Similarly, injection of IL-10 in vivo with LPS decreases the elevated IL-6 serum levels during endotoxemia in Btk-deficient mice. These data further support a role for Btk in regulating TLR-induced cytokine production from APCs and provide downstream targets for analysis of Btk function. PMID- 17082639 TI - A novel role of hypoxia-inducible factor in cobalt chloride- and hypoxia-mediated expression of IL-8 chemokine in human endothelial cells. AB - Tissue hypoxemia is common in several pathological diseases, including vaso occlusion in sickle cell disease and myocardial infarction. One finds increased presence of leukocytes during lung injury and at sites of inflammation in vascular endothelium. In this study, we used human pulmonary microvascular endothelial cells and human dermal microvascular endothelial immortalized cell line to delineate the cellular signaling mechanism of hypoxia- and CoCl2 (a mimetic of hypoxia)-induced IL-8 expression, and the latter's role in chemotaxis of polmorphonuclear neutrophils. We show that hypoxia- and CoCl2-induced IL-8 mRNA and protein expression involved activation of PI3K/Akt and p38 MAPK, but not MEK kinase. Analysis of some transcription factors associated with IL-8 promoter revealed that hypoxia and CoCl2 increased DNA-binding activity of hypoxia inducible factor-1alpha (HIF-1alpha), NF-kappaB, and AP-1. In addition, we show that hypoxia- and CoCl2-induced IL-8 expression requires activation of HIF as demonstrated by the following: 1) EMSA; 2) transfection studies with IL-8 promoter reporter constructs with mutation in HIF-1alpha binding site; 3) attenuation of IL-8 expression by both HIF-1alpha small interfering RNA and R59949; 4) augmentation of IL-8 expression by either transfection with HIF-prolyl hydroxylase-2 small interfering RNA or overexpression of HIF-1alpha; and 5) chromatin immunoprecipitation analysis. Moreover, conditioned medium from hypoxia treated endothelial cells augmented chemotaxis of neutrophils, due to release of IL-8. These data indicate that hypoxia-induced signaling in vascular endothelium for transcriptional activation of IL-8 involves PI3K/Akt, p38 MAPK, and HIF 1alpha. Pharmacological agents, which inhibit HIF-1alpha, may possibly ameliorate inflammation associated with hypoxia in pathological diseases. PMID- 17082640 TI - Urothelial CD44 facilitates Escherichia coli infection of the murine urinary tract. AB - Escherichia coli is the most common pathogen found in urinary tract infections (UTIs), mainly affecting children and women. We report that CD44, a hyaluronic acid (HA) binding protein that mediates cell-cell and cell-matrix interactions, facilitates the interaction of E. coli with urothelial cells and thus the infection of the host. We found that CD44 is constitutively expressed on urothelial cells and that HA accumulates in E. coli-induced UTI. In CD44 deficient mice, the bacterial outgrowth was dramatically less compared with wild type mice despite similar granulocyte influx in the bladder and in the kidney as well as comparable cytokines/chemokines levels in both genotypes. E. coli was able to bind HA, which adhered to CD44-positive tubular epithelial cells. Most importantly, the interaction of CD44 on tubular epithelial cells with HA facilitated the migration of E. coli through the epithelial monolayer. The results provide evidence that CD44 on urothelial cells facilitates E. coli UTI. Disruption of the interaction between CD44 and HA in the bladder may provide a new approach to prevent and to treat UTI. PMID- 17082641 TI - Systemic inhibition of the angiotensin-converting enzyme limits lipopolysaccharide-induced lung neutrophil recruitment through both bradykinin and angiotensin II-regulated pathways. AB - Recruitment of neutrophils to the lung is a sentinel event in acute lung inflammation. Identifying mechanisms that regulate neutrophil recruitment to the lung may result in strategies to limit lung damage and improve clinical outcomes. Recently, the renin angiotensin system (RAS) has been shown to regulate neutrophil influx in acute inflammatory models of cardiac, neurologic, and gastrointestinal disease. As a role for the RAS in LPS-induced acute lung inflammation has not been described, we undertook this study to examine the possibility that the RAS regulates neutrophil recruitment to the lung after LPS exposure. Pretreatment of mice with the angiotensin-converting enzyme (ACE) inhibitor enalapril, but not the anti-hypertensive hydralazine, decreased pulmonary neutrophil recruitment after exposure to LPS. We hypothesize that inhibition of LPS-induced neutrophil accumulation to the lung with enalapril occurred through both an increase in bradykinin, and a decrease in angiotensin II (ATII), mediated signaling. Bradykinin receptor blockade reversed the inhibitory effect of enalapril on neutrophil recruitment. Similarly, pretreatment with bradykinin receptor agonists inhibited IL-8-induced neutrophil chemotaxis and LPS induced neutrophil recruitment to the lung. Inhibition of ATII-mediated signaling, with the ATII receptor 1a inhibitor losartan, decreased LPS-induced pulmonary neutrophil recruitment, and this was suggested to occur through decreased PAI-1 levels. LPS-induced PAI-1 levels were diminished in animals pretreated with losartan and in those deficient for the ATII receptor 1a. Taken together, these results suggest that ACE regulates LPS-induced pulmonary neutrophil recruitment via modulation of both bradykinin- and ATII-mediated pathways, each regulating neutrophil recruitment by separate, but distinct, mechanisms. PMID- 17082642 TI - Metalloproteinases control brain inflammation induced by pertussis toxin in mice overexpressing the chemokine CCL2 in the central nervous system. AB - Inflammatory leukocytes infiltrate the CNS parenchyma in neuroinflammation. This involves cellular migration across various structures associated with the blood brain barrier: the vascular endothelium, the glia limitans, and the perivascular space between them. Leukocytes accumulate spontaneously in the perivascular space in brains of transgenic (Tg) mice that overexpress CCL2 under control of a CNS specific promoter. The Tg mice show no clinical symptoms, even though leukocytes have crossed the endothelial basement membrane. Pertussis toxin (PTx) given i.p. induced encephalopathy and weight loss in Tg mice. We used flow cytometry, ultra small superparamagnetic iron oxide-enhanced magnetic resonance imaging, and immunofluorescent staining to show that encephalopathy involved leukocyte migration across the glia limitans into the brain parenchyma, identifying this as the critical step in inducing clinical symptoms. Metalloproteinase (MPs) enzymes are implicated in leukocyte infiltration in neuroinflammation. Unmanipulated Tg mice had elevated expression of tissue inhibitor of metalloproteinase-1, matrix metalloproteinase (MMP)-10, and -12 mRNA in the brain. PTx further induced expression of tissue inhibitor of metalloproteinase-1, metalloproteinase disintegrins-12, MMP-8, and -10 in brains of Tg mice. Levels of the microglial associated MP MMP-15 were not affected in control or PTx-treated Tg mice. PTx also up-regulated expression of proinflammatory cytokines IL-1beta and TNF-alpha mRNA in Tg CNS. Weight loss and parenchymal infiltration, but not perivascular accumulation, were significantly inhibited by the broad-spectrum MP inhibitor BB 94/Batimastat. Our finding that MPs mediate PTx-induced parenchymal infiltration to the chemokine-overexpressing CNS has relevance for the pathogenesis of human diseases involving CNS inflammation, such as multiple sclerosis. PMID- 17082643 TI - Chronic hyperglycemia predisposes to exaggerated inflammatory response and leukocyte dysfunction in Akita mice. AB - The role of polymorphonuclear neutrophils (PMN) in mediating diabetic tissue damage to the periodontium was investigated in a novel model of chronic hyperglycemia, the Akita mouse. Induction of acute peritoneal inflammation in wild-type (WT) and Akita mice resulted in exaggerated IL-6 response in Akita mice (2.9-fold increase over WT values) and a markedly increased chemokine response (KC, 2.6-fold; MCP-1, 2.6-fold; and MIP-1alpha, 4.4-fold increase over WT values). Chemotaxis to both fMLP and WKYMVm was significantly reduced in isolated Akita PMN compared with WT PMN as measured in a Boyden chamber. Superoxide release in contrast was significantly increased in Akita PMN as measured with cytochrome c reduction. Bone marrow-derived Akita PMN showed partial translocation of p47phox to the cell membrane without external stimulation, suggesting premature assembly of the superoxide-producing NADPH oxidase in hyperglycemia. In vivo studies revealed that ligature-induced periodontal bone loss is significantly greater in Akita mice compared with WT. Moreover, intravital microscopy of gingival vessels showed that leukocyte rolling and attachment to the vascular endothelium is enhanced in periodontal vessels of Akita mice. These results indicate that chronic hyperglycemia predisposes to exaggerated inflammatory response and primes leukocytes for marginalization and superoxide production but not for transmigration. Thus, leukocyte defects in hyperglycemia may contribute to periodontal tissue damage by impairing the innate immune response to periodontal pathogens as well as by increasing free radical load in the gingival microvasculature. PMID- 17082644 TI - Involvement of the purinergic P2X7 receptor in the formation of multinucleated giant cells. AB - Multinucleated giant cells (MGC), a hallmark of chronic inflammatory reactions, remain an enigma of cell biology. There is evidence implicating the purinergic P2X7 receptor in the fusion process leading to MGC. To investigate this, we used HEK 293 cells stably transfected with either 1) the full-length rat P2X7 receptor (P2X7 cells), 2) a rat P2X7 receptor lacking the C-terminal domain (P2X7TC), or 3) a mock vector, and rat alveolar macrophages (MA) expressing the native receptor. P2X7 cells cultured in serum-free medium formed increased numbers of MGC and displayed a higher fusion index compared with mock transfectants. Stimulation of P2X7 pore-forming activity in P2X7 cells by polymyxin B (PMB) further increased significantly the formation of MGC. Conversely, blockers of P2X receptors including oxidized ATP, brilliant blue G, and pyridoxal phosphate-6 azophenyl-2'-4'-disulfonic acid inhibited significantly MGC formation in both unstimulated and PMB-stimulated P2X7-transfected cells. In contrast, cells transfected with the truncated P2X7TC were devoid of pore-forming activity, did not respond to PMB stimulation, and failed to form enhanced numbers of MGC, thus behaving as mock transfectants. As found for P2X7-transfected cells, PMB also potentiated dose-dependently the formation of multinucleated MA by rat alveolar MA. Pretreatment with oxidized ATP abrogated the PMB stimulatory effects. Together, these data demonstrate unequivocally the participation of P2X7 receptor in the process of MGC formation. Our study also provides evidence suggesting that stimulation of the P2X7 receptor pathway in MA may mediate increased formation of MGC during chronic inflammatory reactions. PMID- 17082645 TI - Complement-dependent P-selectin expression and injury following ischemic stroke. AB - The mechanisms that contribute to inflammatory damage following ischemic stroke are poorly characterized, but studies indicate a role for both complement and P selectin. In this study, we show that compared with wild-type mice, C3-deficient mice showed significant improvement in survival, neurological deficit, and infarct size at 24 h after middle cerebral artery occlusion and reperfusion. Furthermore, P-selectin protein expression was undetectable in the cerebral microvasculature of C3-deficient mice following reperfusion, and there was reduced neutrophil influx, reduced microthrombus formation, and increased blood flow postreperfusion in C3-deficient mice. We further investigated the use of a novel complement inhibitory protein in a therapeutic paradigm. Complement receptor 2 (CR2)-Crry inhibits complement activation at the C3 stage and targets to sites of complement activation. Treatment of normal mice with CR2-Crry at 30 min postreperfusion resulted in a similar level of protection to that seen in C3 deficient mice in all of the above-measured parameters. The data demonstrate an important role for complement in cerebrovascular thrombosis, inflammation, and injury following ischemic stroke. P-selectin expression in the cerebrovasculature, which is also implicated in cerebral ischemia and reperfusion injury, was shown to be distal to and dependent on complement activation. Data also show that a CR2-targeted approach of complement inhibition provides appropriate bioavailability in cerebral injury to enable complement inhibition at a dose that does not significantly affect systemic levels of serum complement activity, a potential benefit for stroke patients where immunosuppression would be undesirable due to significantly increased susceptibility to lung infection. PMID- 17082646 TI - Evidence of a functional role for mast cells in the development of type 1 diabetes mellitus in the BioBreeding rat. AB - Human type 1 diabetes mellitus (T1DM) arises through autoimmune destruction of pancreatic beta cells and is modeled in many respects by the lymphopenic and spontaneously diabetic BioBreeding (BB) DRlyp/lyp rat. Previously, preonset expression profiling of whole DRlyp/lyp pancreatic lymph nodes (PLN) revealed innate immune activity, specifically that of mast cells and eosinophils. Furthermore, we observed that pancreatic islets of DRlyp/lyp rats as well as those of diabetes-inducible BB DR(+/+) rats potentially recruit innate cells through eotaxin expression. Here we determine that lifelong eotaxin expression begins before 40 days of life and is localized specifically to beta cells. In this report, we find that PLN mast cells are more abundant in DRlyp/lyp compared with related BB DR(+/+) rats (2.1 +/- 0.9% vs 0.9 +/- 0.4% of total cells, p < 0.0001). DRlyp/lyp PLN mast cell gene expression profiling revealed an activated population and included significant overrepresentation of transcripts for mast cell protease 1, cationic trypsinogen, carboxypeptidase A, IL-5, and phospholipase Cgamma. In the DR(+/+) rat, which develops T1DM upon depletion of T regulator cells, mast cells displayed gene expression consistent with the negative regulation of degranulation, including significant overrepresentation of transcripts encoding tyrosine phosphatase SHP-1, lipid phosphatase SHIP, and E3 ubiquitin ligase c-Cbl. To recapitulate the negative mast cell regulation observed in the DR(+/+) rats, we treated DRlyp/lyp rats with the mast cell "stabilizer" cromolyn, which significantly (p < 0.05) delayed T1DM onset. These findings are consistent with a growing body of evidence in human and animal models, where a role for mast cells in the initiation and progression of autoimmune disease is emerging. PMID- 17082647 TI - FcR-bearing myeloid cells are responsible for triggering murine lupus nephritis. AB - Lupus glomerulonephritis is initiated by deposition of IgG-containing immune complexes in renal glomeruli. FcR engagement by immune complexes (IC) is crucial to disease development as uncoupling this pathway in FcRgamma(-/-) abrogates inflammatory responses in (NZB x NZW)F1 mice. To define the roles of FcR-bearing hemopoietic cells and of kidney resident mesangial cells in pathogenesis, (NZB x NZW)F1 bone marrow chimeras were generated. Nephritis developed in (NZB x NZW)F1 mice expressing activating FcRs in hemopoietic cells. Conversely, recipients of FcRgamma(-/-) bone marrow were protected from disease development despite persistent expression of FcRgamma in mesangial cell populations. Thus, activating FcRs on circulating hemopoietic cells, rather than on mesangial cells, are required for IC-mediated pathogenesis in (NZB x NZW)F1. Transgenic FcRgamma(-/-) mice expressing FcRgamma limited to the CD11b+ monocyte/macrophage compartment developed glomerulonephritis in the anti-glomerular basement disease model, whereas nontransgenic FcRgamma(-/-) mice were completely protected. Thus, direct activation of circulating FcR-bearing myeloid cells, including monocytes/macrophages, by glomerular IC deposits is sufficient to initiate inflammatory responses. PMID- 17082648 TI - The chemokine decoy receptor M3 blocks CC chemokine ligand 2 and CXC chemokine ligand 13 function in vivo. AB - Chemokines and their receptors play a key role in immune homeostasis regulating leukocyte migration, differentiation, and function. Viruses have acquired and optimized molecules that interact with the chemokine system. These virus-encoded molecules promote cell entry, facilitate dissemination of infected cells, and enable the virus to evade the immune response. One such molecule in the murine gammaherpesvirus 68 genome is the M3 gene, which encodes a secreted 44-kDa protein that binds with high affinity to certain murine and human chemokines and blocks chemokine signaling in vitro. To test the hypothesis that M3 directly interferes with diverse chemokines in vivo, we examined the interaction of M3 with CCL2 and CXCL13 expressed in the pancreas of transgenic mice. CCL2 expression in the pancreas promoted recruitment of monocytes and dendritic cells; CXCL13 promoted recruitment of B and T lymphocytes. Coexpression of M3 in the pancreas blocked cellular recruitment induced by both CCL2 and CXCL13. These results define M3 as multichemokine blocker and demonstrate its use as a powerful tool to analyze chemokine biology. PMID- 17082649 TI - Transcriptional profiling of the human monocyte-to-macrophage differentiation and polarization: new molecules and patterns of gene expression. AB - Comprehensive analysis of the gene expression profiles associated with human monocyte-to-macrophage differentiation and polarization toward M1 or M2 phenotypes led to the following main results: 1) M-CSF-driven monocyte-to macrophage differentiation is associated with activation of cell cycle genes, substantiating the underestimated proliferation potential of monocytes. 2) M-CSF leads to expression of a substantial part of the M2 transcriptome, suggesting that under homeostatic conditions a default shift toward M2 occurs. 3) Modulation of genes involved in metabolic activities is a prominent feature of macrophage differentiation and polarization. 4) Lipid metabolism is a main category of modulated transcripts, with expected up-regulation of cyclo-oxygenase 2 in M1 cells and unexpected cyclo-oxygenase 1 up-regulation in M2 cells. 5) Each step is characterized by a different repertoire of G protein-coupled receptors, with five nucleotide receptors as novel M2-associated genes. 6) The chemokinome of polarized macrophages is profoundly diverse and new differentially expressed chemokines are reported. Thus, transcriptome profiling reveals novel molecules and signatures associated with human monocyte-to-macrophage differentiation and polarized activation which may represent candidate targets in pathophysiology. PMID- 17082650 TI - Proteomic identification of in vivo substrates for matrix metalloproteinases 2 and 9 reveals a mechanism for resolution of inflammation. AB - Clearance of allergic inflammatory cells from the lung through matrix metalloproteinases (MMPs) is necessary to prevent lethal asphyxiation, but mechanistic insight into this essential homeostatic process is lacking. In this study, we have used a proteomics approach to determine how MMPs promote egression of lung inflammatory cells through the airway. MMP2- and MMP9-dependent cleavage of individual Th2 chemokines modulated their chemotactic activity; however, the net effect of complementing bronchoalveolar lavage fluid of allergen-challenged MMP2(-/-)/MMP9(-/-) mice with active MMP2 and MMP9 was to markedly enhance its overall chemotactic activity. In the bronchoalveolar fluid of MMP2(-/-)/MMP9(-/-) allergic mice, we identified several chemotactic molecules that possessed putative MMP2 and MMP9 cleavage sites and were present as higher molecular mass species. In vitro cleavage assays and mass spectroscopy confirmed that three of the identified proteins, Ym1, S100A8, and S100A9, were substrates of MMP2, MMP9, or both. Function-blocking Abs to S100 proteins significantly altered allergic inflammatory cell migration into the alveolar space. Thus, an important effect of MMPs is to differentially modify chemotactic bioactivity through proteolytic processing of proteins present in the airway. These findings provide a molecular mechanism to explain the enhanced clearance of lung inflammatory cells through the airway and reveal a novel approach to target new therapies for asthma. PMID- 17082651 TI - Expression and functions of the vascular endothelial growth factors and their receptors in human basophils. AB - Angiogenesis is a multistep complex phenomenon critical for several inflammatory and neoplastic disorders. Basophils, normally confined to peripheral blood, can infiltrate the sites of chronic inflammation. In an attempt to obtain insights into the mechanism(s) underlying human basophil chemotaxis and its role in inflammation, we have characterized the expression and function of vascular endothelial growth factors (VEGFs) and their receptors in these cells. Basophils express mRNA for three isoforms of VEGF-A (121, 165, and 189) and two isoforms of VEGF-B (167 and 186). Peripheral blood and basophils in nasal polyps contain VEGF A localized in secretory granules. The concentration of VEGF-A in basophils was 144.4 +/- 10.8 pg/10(6) cells. Immunologic activation of basophils induced the release of VEGF-A. VEGF-A (10-500 ng/ml) induced basophil chemotaxis. Supernatants of activated basophils induced an angiogenic response in the chick embryo chorioallantoic membrane that was inhibited by an anti-VEGF-A Ab. The tyrosine kinase VEGFR-2 (VEGFR-2/KDR) mRNA was expressed in basophils. These cells also expressed mRNA for the soluble form of VEGFR-1 and neuropilin (NRP)1 and NRP2. Flow cytometric analysis indicated that basophils express epitopes recognized by mAbs against the extracellular domains of VEGFR-2, NRP1, and NRP2. Our data suggest that basophils could play a role in angiogenesis and inflammation through the expression of several forms of VEGF and their receptors. PMID- 17082652 TI - Inhibition of Helicobacter hepaticus-induced colitis by IL-10 requires the p50/p105 subunit of NF-kappa B. AB - Defects within the innate immune system sensitize NF-kappaB-deficient (p50(-/-); p65(+/-)) mice to Helicobacter hepaticus (Hh)-induced colitis. Because IL-10 plays a central role in the inhibition of Hh-induced colitis, we hypothesized that the ability of IL-10 to inhibit the innate inflammatory response to Hh may be compromised in NF-kappaB-deficient mice. To test this hypothesis, we evaluated the ability of an IL-10-Ig fusion protein with IL-10-like properties to inhibit Hh-induced colitis in RAG-2(-/-) (RAG) and p50(-/-); p65(+/-); RAG-2(-/-) (3X/RAG) mice. As expected, IL-10-Ig efficiently inhibited the development of colitis in RAG mice. In contrast, the ability of IL-10-Ig to inhibit colitis was compromised in 3X/RAG mice. The defect in response to IL-10-Ig appeared to be primarily the result of the absence of the p50/p105 subunit, because the ability of IL-10-Ig to inhibit colitis was also compromised in p50(-/-); RAG-2(-/-) (p50/RAG) mice. Radiation chimeras demonstrated that the presence of p50/p105 within hemopoietic cells of the innate immune system was necessary for efficient inhibition of colitis by IL-10-Ig. Consistent with a defect in the suppressive effects of IL-10 in the absence of p50/p105, we found that the ability of IL-10 to control LPS-induced expression of IL-12 p40 was significantly compromised in macrophages lacking p50/p105. These results suggest that the absence of the p50/p105 subunit of NF-kappaB within hemopoietic cells of the innate immune system interferes with the ability of IL-10 to suppress inflammatory gene expression and Hh-induced colitis. PMID- 17082653 TI - Major basic protein homolog (MBP2): a specific human eosinophil marker. AB - Human eosinophil granule major basic protein (MBP1) is an exceedingly basic (isoelectric point >11) 14-kDa protein, comprising the core of the secondary eosinophil granule. Recently, a less cationic homolog of MBP, termed MBPH or simply, MBP2, has been discovered. We prepared a panel of mAbs to MBP2 and used these Abs to localize and quantitate this molecule in leukocytes and biological fluids. Specific mAbs for MBP2 were selected using slot-blot analyses and used in a two-site immunoassay, Western blotting, and immunofluorescence microscopy. The sensitivity of the immunoassay was markedly improved by reduction and alkylation of MBP2. MBP1 is more abundant than MBP2 in lysates of eosinophils and their granules, as judged by immunoassay and Western blotting. By immunofluorescence, MBP1 is present in eosinophils, basophils, and a human mast cell line (HMC1), whereas MBP2 is only detected in eosinophils. Neither MBP1 nor MBP2 could be detected in any other peripheral blood leukocyte. MBP2 levels measured in plasma and serum were essentially identical. In contrast to past measurements for MBP1, MBP2 was not detected above normal levels in sera from pregnant donors. However, measurement of serum MBP2 discriminated patients with elevated eosinophils from normal subjects, and MBP2 was also detectable in other biological specimens, such as bronchoalveolar lavage, sputum, and stool. These results indicate that MBP2 is present only in eosinophils and that it may be a useful biomarker for eosinophil associated diseases. PMID- 17082654 TI - Induction of tolerance to innocuous inhaled antigen relies on a CCR7-dependent dendritic cell-mediated antigen transport to the bronchial lymph node. AB - Allergic airway diseases such as asthma are caused by a failure of the immune system to induce tolerance against environmental Ags. The underlying molecular and cellular mechanisms that initiate tolerance are only partly understood. In this study, we demonstrated that a CCR7-dependent migration of both CD103+ and CD103- lung dendritic cells (DC) to the bronchial lymph node (brLN) is indispensable for this process. Although inhaled Ag is amply present in the brLN of CCR7-deficient mice, T cells cannot be tolerized because of the impaired migration of Ag-carrying DC and subsequent transport of Ag from the lung to the draining lymph node. Consequently, the repeated inhalation of Ag protects wild type but not CCR7-deficient mice from developing allergic airway diseases. Thus, the continuous DC-mediated transport of inhaled Ag to the brLN is critical for the induction of tolerance to innocuous Ags. PMID- 17082655 TI - Porcine endothelial cells and iliac arteries transduced with AdenoIL-4 are intrinsically protected, through Akt activation, against immediate injury caused by human complement. AB - Vascular endothelial cells (ECs) can be injured in a variety of pathologic processes that involve activated complement. We reported previously that porcine ECs incubated with exogenous IL-4 or IL-13 are protected from cytotoxicity by human complement and also from apoptosis by TNF-alpha. The resistance to complement consists of an intrinsic mechanism that is lost a few days after cytokine removal. In our current study, we investigated whether transfer of the IL-4 gene into porcine ECs in vitro and into porcine vascular tissues in vivo would induce efficient and durable protection from human complement. We found that ECs transduced with adenoIL-4 or adenoIL-13 exhibited continuous production of the cytokine and prolonged protection from complement-mediated killing. IL-4 also protected ECs from activation: ECs incubated with IL-4 did not develop cell retraction and intercellular gaps upon stimulation with sublytic complement. The endothelium and subendothelium of pig iliac arteries that were transduced with the IL-4 gene were effectively protected from complement-dependent immediate injury after perfusion with human blood. However, after similar perfusion, the endothelium was immediately lost from arteries that were transduced with a control adenovirus. The protection was not due to up-regulation of the complement regulators decay accelerating factor, membrane cofactor protein, and CD59, or to reduced complement activation, but required the participation of Akt. Although our studies model protection in pig-to-primate xenotransplantation, our findings of IL-4 induction of Akt-mediated protection may be more broadly applicable to EC injury as manifested in ischemia-reperfusion, allotransplantation, and various vascular diseases. PMID- 17082656 TI - Epitope specificity of autoreactive T and B cells associated with experimental autoimmune encephalomyelitis and optic neuritis induced by oligodendrocyte specific protein in SJL/J mice. AB - The encephalitogenic potential of oligodendrocyte-specific protein (OSP) in mice, its specific localization in the intralamellar tight junctions in CNS myelin, and the detection of autoreactivity against OSP in multiple sclerosis (MS) strongly suggest the relevance of autoreactivity against OSP in the pathogenesis of MS. In this study, we have characterized the autoimmune T and B cells that are associated with clinicopathological manifestations of OSP-induced MS-like disease in mice by using recombinant soluble mouse OSP (smOSP) and synthetic overlapping peptides spanning smOSP. SJL/J mice immunized with smOSP developed chronic relapsing clinical experimental autoimmune encephalomyelitis accompanied with intense perivascular and parenchymal inflammatory infiltrates, widespread demyelination, axonal loss, and remarkable optic neuritis. The smOSP-primed lymph node cells reacted predominantly against OSP55-80 and to a lesser extent also to OSP22-46 and OSP179-207. Unexpectedly, in vitro selection with smOSP resulted in pathogenic smOSP-specific CD4+ T cells that reacted equally well against OSP55 80, OSP22-46, OSP45-66, and OSP179-207. Fine analysis of the anti-OSP autoimmunity revealed that the disease is primarily associated with CD4+ T cells directed against the major (OSP55-80) and the minor (OSP179-207) encephalitogenic regions that were further delineated, both in vitro and in vivo, to OSP55-66 and OSP194-207, respectively. In contrast, the OSP-induced Abs were predominantly directed against OSP22-46; these Abs were mostly of IgG1 isotype, but high levels of IgG2a and IgG2b and significant levels of IgE were also observed. The reactivity of pathogenic T cells to two encephalitogenic regions, OSP55-80 and OSP179-207, and their diverse TCRVbeta gene repertoire may impose difficulties for epitope-directed or TCR-targeting approaches to immune-specific modulation of OSP-related pathogenesis. PMID- 17082657 TI - A single amino acid difference within the alpha-2 domain of two naturally occurring equine MHC class I molecules alters the recognition of Gag and Rev epitopes by equine infectious anemia virus-specific CTL. AB - Although CTL are critical for control of lentiviruses, including equine infectious anemia virus, relatively little is known regarding the MHC class I molecules that present important epitopes to equine infectious anemia virus specific CTL. The equine class I molecule 7-6 is associated with the equine leukocyte Ag (ELA)-A1 haplotype and presents the Env-RW12 and Gag-GW12 CTL epitopes. Some ELA-A1 target cells present both epitopes, whereas others are not recognized by Gag-GW12-specific CTL, suggesting that the ELA-A1 haplotype comprises functionally distinct alleles. The Rev-QW11 CTL epitope is also ELA-A1 restricted, but the molecule that presents Rev-QW11 is unknown. To determine whether functionally distinct class I molecules present ELA-A1-restricted CTL epitopes, we sequenced and expressed MHC class I genes from three ELA-A1 horses. Two horses had the 7-6 allele, which when expressed, presented Env-RW12, Gag GW12, and Rev-QW11 to CTL. The other horse had a distinct allele, designated 141, encoding a molecule that differed from 7-6 by a single amino acid within the alpha-2 domain. This substitution did not affect recognition of Env-RW12, but resulted in more efficient recognition of Rev-QW11. Significantly, CTL recognition of Gag-GW12 was abrogated, despite Gag-GW12 binding to 141. Molecular modeling suggested that conformational changes in the 141/Gag-GW12 complex led to a loss of TCR recognition. These results confirmed that the ELA-A1 haplotype is comprised of functionally distinct alleles, and demonstrated for the first time that naturally occurring MHC class I molecules that vary by only a single amino acid can result in significantly different patterns of epitope recognition by lentivirus-specific CTL. PMID- 17082658 TI - Severe focal sialadenitis and dacryoadenitis in NZM2328 mice induced by MCMV: a novel model for human Sjogren's syndrome. AB - The genetic and environmental factors that control the development of Sjogren's syndrome, an autoimmune disease mainly involving the salivary and lacrimal glands, are poorly understood. Viruses which infect the glands may act as a trigger for disease. The ability of sialotropic murine CMV (MCMV) to induce acute and chronic glandular disease was characterized in an autoimmune-prone mouse strain, NZM2328. MCMV levels were detectable in the salivary and lacrimal glands 14-28 days after i.p. infection and correlated with acute inflammation in the submandibular gland. After latency, virus was undetectable in the glands by PCR. At this stage, NZM2328 female mice developed severe chronic periductal inflammation in both submandibular and lacrimal glands in contrast to the much milder infiltrates found in female B6-lpr and male NZM2328. The focal infiltrates consisted of CD4+ and B220+ cells as opposed to diffuse CD4+, CD8+, and B220+ cells during acute infection. Salivary gland functional studies revealed a gender specific progressive loss of secretory function between days 90 and 125 postinfection. Latent MCMV infection did not significantly affect the low incidence of autoantibodies to Ro/SSA and La/SSB Ags in NZM2328 mice. However, reactivities to other salivary and lacrimal gland proteins were readily detected. MCMV infection did not significantly alter the spontaneous onset of kidney disease in NZM2328. Thus, chronic inflammation induced by MCMV with decreased secretory function in NZM2328 mice resembles the disease manifestations of human Sjogren's syndrome. PMID- 17082659 TI - CD4+CD25high T cells are enriched in the tumor and peripheral blood of prostate cancer patients. AB - In this study, we investigated whether CD4+CD25high regulatory T cells (Treg) are increased in the tumor tissue and peripheral blood of early-stage prostate cancer patients undergoing prostatectomy. We show that the prevalence of CD4+CD25high T cells inside the prostate was significantly higher in the tumor compared with benign tissue from the same prostate. Furthermore, the frequency of CD4+CD25high T cells in peripheral blood was significantly higher in prostate cancer patients compared with normal donors. A proportion of the CD4+CD25high T cells was also shown to be glucocorticoid-induced TNF receptor, ICOS, and FOXP3 positive. Moreover, CD4+CD25+ T cells from blood and supernatants from cultured prostate tumor tissue samples exhibited immunosuppressive function in vitro. Furthermore, supernatants from cultured prostate tissue samples and prostate cancer ascites fluid induced migration of CD4+CD25+ T cells and were shown to contain the regulatory T cell chemokine CCL22 by ELISA. Our findings indicate that Tregs are an important cellular component of early-stage prostate tumors, and thus new therapeutic strategies aimed at inhibition or depletion of Tregs may improve prostate cancer immunotherapy. PMID- 17082660 TI - Preservation of T cell proliferation restricted by protective HLA alleles is critical for immune control of HIV-1 infection. AB - HIV-1-infected persons with HLA-B27 and -B57 alleles commonly remain healthy for decades without antiretroviral therapy. Properties of CD8+ T cells restricted by these alleles considered to confer disease protection in these individuals are elusive but important to understand and potentially elicit by vaccination. To address this, we compared CD8+ T cell function induced by HIV-1 immunogens and natural infection using polychromatic flow cytometry. HIV-1-specific CD8+ T cells from all four uninfected immunized and 21 infected subjects secreted IFN-gamma and TNF-alpha. However, CD8+ T cells induced by vaccination and primary infection, but not chronic infection, proliferated to their cognate epitopes. Notably, B27- and B57-restricted CD8+ T cells from nonprogressors exhibited greater expansion than those restricted by other alleles. Hence, CD8+ T cells restricted by certain protective alleles can resist replicative defects, which permits expansion and antiviral effector activities. Our findings suggest that the capacity to maintain CD8+ T cell proliferation, regardless of MHC restriction, may serve as an important correlate of disease protection in the event of infection following vaccination. PMID- 17082661 TI - Atorvastatin restores Lck expression and lipid raft-associated signaling in T cells from patients with systemic lupus erythematosus. AB - Loss of tolerance to self-Ags in patients with systemic lupus erythematosus (SLE), a prototypic autoimmune disease, is associated with dysregulation of T cell signaling, including the depletion of total levels of lymphocyte-specific protein kinase (Lck) from sphingolipid-cholesterol-enriched membrane microdomains (lipid rafts). Inhibitors of 3-hyroxy-3-methylgluteryl CoA reductase (statins) can modify the composition of lipid rafts, resulting in alteration of T cell signaling. In this study, we show that atorvastatin targets the distribution of signaling molecules in T cells from SLE patients, by disrupting the colocalization of total Lck and CD45 within lipid rafts, leading to a reduction in the active form of Lck. Upon T cell activation using anti-CD3/anti-CD28 in vitro, the rapid recruitment of total Lck to the immunological synapse was inhibited by atorvastatin, whereas ERK phosphorylation, which is decreased in SLE T cells, was reconstituted. Furthermore, atorvastatin reduced the production of IL-10 and IL-6 by T cells, implicated in the pathogenesis of SLE. Thus, atorvastatin reversed many of the signaling defects characteristic of SLE T cells. These findings demonstrate the potential for atorvastatin to target lipid raft-associated signaling abnormalities in autoreactive T cells and provide a rationale for its use in therapy of autoimmune disease. PMID- 17082662 TI - Role of MHC-linked genes in autoantigen selection and renal disease in a murine model of systemic lupus erythematosus. AB - We previously described a renal protective effect of factor B deficiency in MRL/lpr mice. Factor B is in the MHC cluster; thus, the deficient mice were H2b, the haplotype on which the knockout was derived, whereas the wild-type littermates were H2k, the H2 of MRL/lpr mice. To determine which protective effects were due to H2 vs factor B deficiency, we derived H2b congenic MRL/lpr mice from the 129/Sv (H2b) strain. Autoantibody profiling using autoantigen microarrays revealed that serum anti-Smith and anti-small nuclear ribonucleoprotein complex autoantibodies, while present in the majority of H2k/k MRL/lpr mice, were absent in the H2b/b MRL/lpr mice. Surprisingly, 70% of MRL/lpr H2b/b mice were found to be serum IgG3 deficient (with few to no IgG3-producing B cells). In addition, H2b/b IgG3-deficient MRL/lpr mice had significantly less proteinuria, decreased glomerular immune complex deposition, and absence of glomerular subepithelial deposits compared with MRL/lpr mice of any H2 type with detectable serum IgG3. Despite these differences, total histopathologic renal scores and survival were similar among the groups. These results indicate that genes encoded within or closely linked to the MHC region regulate autoantigen selection and isotype switching to IgG3 but have minimal effect on end-organ damage or survival in MRL/lpr mice. PMID- 17082663 TI - Thrice-weekly low-dose rituximab decreases CD20 loss via shaving and promotes enhanced targeting in chronic lymphocytic leukemia. AB - Treatment of chronic lymphocytic leukemia (CLL) patients with standard dose infusion of rituximab (RTX), 375 mg/m2, induces clearance of malignant cells from peripheral blood after infusion of 30 mg of RTX. After completion of the full RTX infusion, substantial recrudescence of CLL cells occurs, and these cells have lost > 90% of CD20. To gain insight into mechanism(s) of CD20 loss, we investigated the hypothesis that thrice-weekly low-dose RTX (20 or 60 mg/m2) treatment for CLL over 4 wk would preserve CD20 and enhance leukemic cell clearance. During initial infusions in all 12 patients, the first 30 mg of RTX promoted clearance of > 75% leukemic cells. Four of six patients receiving 20 mg/m2 RTX retained > or = 50% CD20, and additional RTX infusions promoted further cell clearance. However, four of six patients receiving 60 mg/m2 had CD20 levels < 20% baseline 2 days after initial infusions, and additional RTX infusions were less effective, presumably due to epitope loss. Our results suggest that when a threshold RTX dose is exceeded, recrudesced RTX-opsonized cells are not cleared, due to saturation of the mononuclear phagocytic system, but instead are shaved of RTX-CD20 complexes by acceptor cells. Thrice-weekly low-dose RTX may promote enhanced clearance of circulating CLL cells by preserving CD20. PMID- 17082664 TI - Modulation of autoimmunity by TLR9 in the chronic graft-vs-host model of systemic lupus erythematosus. AB - Chronic graft-vs-host (cGVH) disease is induced in nonautoimmune mice by the transfer of alloreactive T cells that recognize foreign MHC class II. It closely resembles systemic lupus erythematosus, with antinuclear Abs and immune-mediated nephritis. Recent work has implicated TLRs, particularly TLR9, in the recognition of certain autoantigens in vitro and in vivo. To explore further the role of TLR9 in systemic autoimmunity, we induced cGVH disease in C57BL/6 (B6) mice lacking TLR9, including B6 mice expressing the anti-DNA-encoding IgH transgenes 3H9 or 56R (B6.3H9.TLR9(-/-), B6.56R.TLR9(-/-)). We found that cGVH disease caused breakdown of B cell tolerance to chromatin and DNA in TLR9(-/-) recipients of alloreactive cells, yet that nephritis was less severe and that some autoantibody titers were lower compared with B6-cGVH controls. Spleen lymphocyte analysis showed that cGVH disease strikingly depleted marginal zone B cells in B6 mice, but did not influence T cell subsets in either B6 or B6-TLR9(-/-) hosts. B6.56R.TLR9(-/-) mice had less spontaneous production of autoantibodies than B6.56R mice, but there were no significant differences between B6.56R and B6.56R.TLR9(-/-) postinduction of cGVH disease. Taken together, these results suggested that TLR9 may worsen some aspects of systemic autoimmunity while alleviating others. PMID- 17082665 TI - Defective Th1 cytokine gene transcription in CD4+ and CD8+ T cells from Wiskott Aldrich syndrome patients. AB - Wiskott-Aldrich syndrome (WAS) protein (WASP) plays a key role in TCR-mediated activation and immunological synapse formation. However, the effects of WASP deficiency on effector functions of human CD4+ and CD8+ T cells remain to be determined. In this study, we report that TCR/CD28-driven proliferation and secretion of IL-2, IFN-gamma, and TNF-alpha are strongly reduced in CD8+ T cells from WAS patients, compared with healthy donor CD8+ T cells. Furthermore, WAS CD4+ T cells secrete low levels of IL-2 and fail to produce IFN-gamma and TNF alpha, while the production of IL-4, IL-5, and IL-10 is only minimally affected. Defective IL-2 and IFN-gamma production persists after culture of naive WAS CD4+ T cells in Th1-polarizing conditions. The defect in Th1 cytokine production by WAS CD4+ and CD8+ T cells is also present at the transcriptional level, as shown by reduced IL-2 and IFN-gamma mRNA transcripts after TCR/CD28 triggering. The reduced transcription of Th1 cytokine genes in WAS CD4+ T cells is associated with a defective induction of T-bet mRNA and a reduction in the early nuclear recruitment of NFAT-1, while the defective activation of WAS CD8+ T cells correlates with reduced nuclear recruitment of both NFAT-1 and NFAT-2. Together, our data indicate that WASP regulates the transcriptional activation of T cells and is required specifically for Th1 cytokine production. PMID- 17082666 TI - Modulation of the immune response induced by gene electrotransfer of a hepatitis C virus DNA vaccine in nonhuman primates. AB - Induction of multispecific, functional CD4+ and CD8+ T cells is the immunological hallmark of acute self-limiting hepatitis C virus (HCV) infection in humans. In the present study, we showed that gene electrotransfer (GET) of a novel candidate DNA vaccine encoding an optimized version of the nonstructural region of HCV (from NS3 to NS5B) induced substantially more potent, broad, and long-lasting CD4+ and CD8+ cellular immunity than naked DNA injection in mice and in rhesus macaques as measured by a combination of assays, including IFN-gamma ELISPOT, intracellular cytokine staining, and cytotoxic T cell assays. A protocol based on three injections of DNA with GET induced a substantially higher CD4+ T cell response than an adenovirus 6-based viral vector encoding the same Ag. To better evaluate the immunological potency and probability of success of this vaccine, we have immunized two chimpanzees and have compared vaccine-induced cell-mediated immunity to that measured in acute self-limiting infection in humans. GET of the candidate HCV vaccine led to vigorous, multispecific IFN-gamma+CD8+ and CD4+ T lymphocyte responses in chimpanzees, which were comparable to those measured in five individuals that cleared spontaneously HCV infection. These data support the hypothesis that T cell responses elicited by the present strategy could be beneficial in prophylactic vaccine approaches against HCV. PMID- 17082669 TI - The evolution of uterine surgery. AB - Although described in antiquity, the real dawn of uterine surgery was in the mid 19th century when hysterectomy was occasionally performed vaginally, usually for cancer or prolapse. Then, as now, women experienced symptoms of bleeding and pain emanating from the uterus, and when severe and debilitating, brave surgeons and patients sometimes explored hysterectomy as an alternative. Abdominal hysterectomy mortality rates in the mid-1850s were extremely high, but reduced drastically in the early to mid-20th century. By the 1950s, total hysterectomy supplanted supracervical techniques, largely as a method for preventing carcinoma of the cervix. Surgical alternatives to hysterectomy started in the 1930s with abdominal myomectomy and the first publication of nonhysteroscopic endometrial ablation from Germany, but by the end of the 20th century, included a plethora of techniques including laparoscopic, hysteroscopic, and interventional radiologic approaches. The advent of early detection of, and even prevention of, preinvasive cervical neoplasia, has led to a reevaluation of the need for total hysterectomy in many patients. In the early years of the millennium, targeted leiomyoma therapy was under development with a range of energy sources including cryogenic and radiofrequency probes, as well as focused ultrasound, targeted and controlled by magnetic resonance imaging. PMID- 17082670 TI - Hysterectomy in the 21st century: different approaches, different challenges. AB - The final decision to perform a certain method of hysterectomy customarily mirrors experience and level of comfort with a particular surgical approach in the context of the patient's condition and indication for surgery. Given the morbidity and recovery associated with a laparotomic incision, every effort should be made to avoid abdominal hysterectomy. The best available evidence points to the advantage of the vaginal approach over other methods of hysterectomy for benign conditions. Regrettably, the state of education in residency programs is not providing a level of surgical competency to meet this charge. Whenever vaginal surgery is not an option, laparoscopically assisted hysterectomy offers the best alternative. Although the promises of supracervical hysterectomy have yet to be demonstrated, laparoscopic supracervical hysterectomy may offer the least morbid alternative to vaginal hysterectomy. PMID- 17082671 TI - Endometrial ablation: where have we been? Where are we going? AB - Endometrial ablation (EA) is targeted destruction of the endothelial surface of the uterine cavity. The procedure was originally designed as a less invasive alternative to hysterectomy for the symptom of heavy menstrual bleeding unrelated to structural pathology of the uterus, that was not responsive to medical therapy. More recently it has become apparent that the procedure can be performed in the presence of submucous leiomyomas, providing they meet a number of size and location criteria. The first EA serie as published in Germany in the 1930s, but the procedure did not attract much attention until the latter part of the 20th century. Currently, EA can be performed under endoscopic direction with the neodymium:yttrium alumnum garnet laser, with a radiofrequency resectoscope, or with an expanding array of nonresectoscopic EA systems. It is apparent that most but not all of the complications associated with resectoscopic endometrial ablation are eliminated with nonresectoscopic endometrial ablation, but serious morbidity has been reported with all of the newer systems to date. Success and patient satisfaction seem to be enduring in the majority of well-selected patients treated in clinical trials, but repeat surgery, usually hysterectomy, is performed in 25% to 40% by 5 years after surgery. Increased efficiencies should be realized if the procedure could be moved to an office setting. PMID- 17082672 TI - Septums and synechiae: approaches to surgical correction. AB - If the Mullerian ducts fail to fuse, or, if the wall which is the result of fusion is not adequately resorbed, the result is a spectrum of uterine abnormalities called Mullerian fusion and absorption defects. The impact of these abnormalities on fertility is a subject for debate, but at least a subset seems to have a negative impact on reproductive performance manifesting in recurrent abortion and/or premature labor. Previous surgical interventions required laparotomy, but, with careful application of imaging techniques, a group of patients can be identified with a uterine septum amenable to removal under hysteroscopic direction with little morbidity. Intrauterine adhesions or synechiae are usually secondary to curettage in the context of missed abortion or pregnancy-related hemorrhage. These lesions cover a spectrum that ranges from minor and insignificant to severe cohesive adhesions that affect menstrual function and fertility. Surgical repair of the endometrial cavity affected with such adhesions presents a challenge to the hysteroscopic surgeon. Appropriate management is controversial but may include second loop hysteroscopy and the use of postoperative adjuvants such as systemic estrogens and intrauterine devices or systems designed to impede the development of adhesions. PMID- 17082673 TI - Laparoscopic myomectomy and abdominal myomectomy. AB - Most women develop myomas during their lifetimes; however, 80% are asymptomatic. When symptoms are determined to be caused by myomas, a number of management options exist that include "watchful waiting," medical therapy, surgery, or more recently uterine artery embolization and focused ultrasound. Myomectomy, either abdominal or laparoscopic, is an approach particularly suited for those women who wish future fertility. It seems clear that, in well trained and experienced hands, well-selected patients can have myomectomy performed under laparoscopic direction. Very large myomas are not as suitable for the laparoscopic approach, but are amenable to a uterine conserving procedure via laparotomy that is facilitated by a number of preoperative and intraoperative measures aimed to minimize or replace operative blood loss. These techniques should provide selected women a uterine conserving procedure with reduced morbidity. PMID- 17082674 TI - Uterine artery occlusion: what is the evidence? AB - Symptomatic uterine fibroids are a relatively common gynecologic condition. In the past, fibroids were exclusively treated by myomectomy and/or hysterectomy. With the advent of uterine artery embolization or uterine artery occlusion, there now exist minimally invasive approaches to fibroid therapy especially for women in whom surgery is contraindicated or for those who wish to retain their uterus and possibly fertility. Fertility and pregnancy outcomes after these minimally invasive therapies are currently being evaluated. PMID- 17082675 TI - Hysteroscopic treatment of submucous myomas. AB - Submucous myomas can cause heavy bleeding, pain, and infertility. Sonohysterography and office hysteroscopy can provide accurate diagnosis. Classification systems have been developed to help predict the difficulty of resection. Patient selection and proper surgical technique are essential to achieve good results and reduce the risk of complications. Hysteroscopic treatment can in many cases provide excellent results and eliminate the need for major surgery. PMID- 17082676 TI - Myolysis of uterine fibroids: is there a role? AB - Myolysis is among the new procedures under development for the treatment of symptoms related to uterine leiomyoma. The procedure targets the destruction of fibroids using one of a number of focused energy delivery systems including those based upon radiofrequency electricity, supercooled cryoprobes, and, most recently, focused ultrasound monitored by real time magnetic resonance imaging. For thermomyolysis and cryomyolysis, delivery of the energy requires access to the tissue by laparoscopy, and, in some instances, hysteroscopy. For focused ultrasound, the patient is detached from the energy source, which is delivered by an array of external beams. Clinical evaluation has been confined to case series, but it is evident that the approach results in a variable degree of reduction of the total uterine mass, and, usually, a reduction in uterine bleeding. Clearly, longer term appropriately designed comparative trials are required that evaluate and compare myolysis with myomectomy, uterine artery embolization, and hysterectomy, to name a few. PMID- 17082678 TI - Pregnancy-associated changes in the hemostatic system. AB - Pregnancy, from implantation to parturition, presents unique and profound challenges to a women's hemostatic system. During pregnancy, potentially catastrophic bleeding can occur during implantation and endovascular trophoblast invasion of the maternal spiral arteries. The risk of hemorrhage reaches a peak during the third stage of labor when the placenta is shorn from the decidua basalis exposing 120 spiral arteries largely denuded of their smooth muscle, and thus, their ability to constrict in response to injury. These challenges are met by dramatic changes in the local uterine, and systemic hemostatic systems. The net effect of these changes is to increase the efficiency of clotting and to impair fibrinolysis. Unfortunately, they also lead to an increase in the prevalence of venous thromboembolism, which is otherwise uncommon in reproductive age women. PMID- 17082679 TI - Laboratory tests for the diagnosis of thrombotic disorders. AB - Diagnosis of thrombotic disorders depends on the performance of a panel of laboratory tests for individual components of the coagulation mechanism. Although genetic tests are simple and accurate, most disorders still require functional or immunologic tests for diagnosis. Tests for the most important thrombotic defects are described. These tests may be significantly altered by pregnancy, hormones, thrombotic events, systemic illness, and anticoagulant therapy. These changes make some thrombotic disorders difficult to diagnose during pregnancy. Testing before pregnancy for women with a personal or family history of thrombosis will simplify the interpretation. PMID- 17082680 TI - Thrombophilia and adverse maternal-perinatal outcome. AB - A recent review of the literature on thrombophilia and adverse pregnancy outcome (APO) reveals contradictory findings. We have limited our review of literature mostly to the most recent decade. On the basis of our review, screening for thrombophilias with a history of APO (preeclampsia, abruptio placenta, intrauterine growth restriction, and fetal loss) is not clear. There are retrospective and prospective studies that recommend testing for genetic and acquired markers of thrombophilia for those with the enumerated APO. The rationale for such recommendation is to use heparin prophylaxis in subsequent pregnancies. However, this recommendation is not based on randomized trials. Hence, a randomized double-blinded controlled trial is urgently needed to evaluate the benefit of heparin during pregnancy in women with a history of APO in association with thrombophilia. PMID- 17082681 TI - Antiphospholipid syndrome and thrombosis. AB - The antiphospholipid syndrome is an autoimmune condition in which venous or arterial thrombosis is a primary clinical feature. The other primary clinical feature is adverse pregnancy outcome, specifically recurrent miscarriage, fetal death, or preterm delivery due to severe preeclampsia or placental insufficiency. The diagnostic autoantibodies for antiphospholipid syndrome are lupus anticoagulant, anticardiolipin, or anti-beta2-glycoprotein I. PMID- 17082682 TI - Thrombophilias, perinatal stroke, and cerebral palsy. AB - Perinatal ischemic stroke has become recognized as a not-rare adverse outcome of pregnancy and a common cause of chronic neurologic disability in children. This review discusses the clinical entity, perinatal stroke, and its relationship to thrombophilias, inherited and acquired, and to maternal and pregnancy factors. PMID- 17082683 TI - Thrombophilia and placental pathology. AB - Thrombophilic states have been associated with a variety of adverse pregnancy outcomes. The underlying placental pathology linking thrombophilia to these outcomes closely resembles that seen in other pregnancy disorders associated with chronic obstruction of the maternal or fetal vasculature. No single placental lesion is pathognomonic for thrombophilia. However, the finding of typical pathologic lesions in the context of recurrent pregnancy loss, severe early onset disease, or neonatal coagulation abnormalities should prompt the consideration of an underlying thrombophilic state. PMID- 17082684 TI - The use of unfractionated heparin and low molecular weight heparins in pregnancy. AB - Currently unfractionated heparin (UH) and low molecular weight heparins (LMWH) are the agents of choice for anticoagulation in pregnancy. LMWH have been used safely without monitoring in nonpregnant patients; however, because of documented changes in the pharmacokinetics of these agents in pregnancy, monitoring with anti-Xa levels is necessary in pregnancy to maintain target therapeutic ranges. Patients requiring only prophylaxis during pregnancy with either UH or LMWH might benefit from occasional assessment of anti-Xa levels to confirm that target prophylactic ranges are being achieved. Although LMWH may cause less osteoporosis than UH at therapeutic doses, the incidence of heparin-induced osteoporosis seems to be low when only prophylactic dosing is used and therefore LMWH do not seem to offer this advantage at low doses. Experience with newer agents such as pentasaccharide inhibitors and direct thrombin inhibitors are limited in pregnancy and it remains to be seen what role these agents will play in women who require anticoagulation in pregnancy. PMID- 17082685 TI - Preconception counseling for women with thrombophilia. AB - Inherited thrombophilias are a heterogeneous group of coagulation disorders that predispose individuals to thromboembolic events. This group of conditions is the major risk factor for thromboembolism during pregnancy and the puerperium. In addition, thrombophilias have been associated with several adverse obstetric events, including pregnancy loss, preeclampsia, placental abruption, and intrauterine growth restriction. An increased risk for these obstetric complications has prompted many authorities to recommend screening and treating pregnant women for thrombophilias. Optimal obstetric management, however, is controversial as thrombophilias are common and many affected individuals are asymptomatic. Indeed, pregnancy outcome in most women with thrombophilias is normal. The most commonly identified inherited thrombophilias are the factor V Leiden and prothrombin G20210A gene mutations. More rare thrombophilias include protein C and S deficiencies, antithrombin III deficiency. Although relatively common, the association between hyperhomocysteinemia and associated mutations (such as the C677 T methylenetetrahydro-folate reductase) and obstetric complications is controversial. PMID- 17082687 TI - Multislice computed tomography in planning nephron-sparing surgery in a prospective study with 76 patients: comparison of radiological and histopathological findings in the infiltration of renal structures. AB - OBJECTIVE: The aim of this prospective study is to determine the diagnostic accuracy of multidetector-row computed tomography (CT) compared to histopathologic findings in tumor staging of renal cell carcinoma, with the focus on tumor stage, vein and artery infiltration, and infiltration of the renal pelvis. MATERIALS AND METHODS: In a prospective study, a total of 76 consecutive patients with suspected renal cell carcinoma were preoperatively assessed for tumor staging using multidetector-row CT. Triphasic CT imaging (i.e., noncontrast, arterial, and parenchymal phase) was performed using multidetector row CT with reconstructed slice thicknesses of 2 and 5 mm. Two blinded readers evaluated the CT scans independently; the results were then correlated with the histopathologic results. RESULTS: A total of 56 renal cell carcinomas were proven on histopathology. Readers 1 and 2 reached a sensitivity of 1.0 and 1.0 and a specificity of 0.41 and 0.42 for arterial infiltration, a sensitivity of 1.0 and 0.86 and a specificity of 0.58 and 0.5 for venous infiltration, and a sensitivity of 0.75 and 1.0 and a specificity of 0.5 and 0.44 for infiltration of the renal pelvis. The correlation between both readers was 0.7 for all modalities. CONCLUSIONS: The multiplanar reconstruction capability of multidetector-row CT allowed good sensitivity in predicting arterial infiltration. The lowest specificity was reached in excluding infiltration of the renal pelvis. Despite its high temporal and spatial resolution, the capacity of multidetector CT to predict intrarenal infiltrations is still limited. PMID- 17082688 TI - CT findings of clear cell carcinoma of the ovary. AB - PURPOSE: The purpose of this article was to retrospectively evaluate computed tomographic (CT) findings in patients with pathologically proved clear cell carcinoma of the ovary. METHODS: CT scans of seven patients (age range, 28-79 years; mean age, 46 years) with nine lesions of histologically proved primary ovarian clear cell carcinoma of the ovary were retrospectively evaluated by two radiologists in consensus. Scans were evaluated for the laterality, size, mass characteristic, margin, attenuation of the cystic portion, internal architecture, and presence of lumen protruding mass. RESULTS: Tumors were unilateral in five patients. The average size of the tumors was 8.6 cm (range, 2.6-12.1 cm). All the masses showed mainly cystic appearance. All nine masses had smooth margin. Average attenuation of the cystic portion of the masses was 24.2 HU (Hounsfield unit) (range, 13-34 HU). Tumors usually appeared as unilocular cystic mass (n=6). Lumen protruding masses appeared in seven tumors. CONCLUSION: On CT scans, primary ovarian clear cell carcinomas appeared as large unilocular, mainly cystic, smooth marginated mass with lumen protruding solid portion and high attenuated cystic portion. PMID- 17082689 TI - Advanced gastric carcinoma with signet ring cell carcinoma versus non-signet ring cell carcinoma: differentiation with multidetector CT. AB - OBJECTIVE: The purpose of this study is to assess the capability of multidetector computed tomography (MDCT) to assist in the differentiation of advanced gastric carcinoma with signet ring cell type from that with non-signet ring cell carcinoma (NSRC) with a focus on the thickened stomach wall itself. METHODS: We retrospectively reviewed MDCT results in 80 patients with pathologically proven advanced gastric carcinoma with signet ring cell carcinoma (SRC) (n = 35) and NSRC (n = 45). MDCT images of 80 patients were analyzed retrospectively on gross appearance of thickened gastric wall (polypoid/fungating/ulcerative/diffuse infiltrative), predominantly thickened layer (inner/outer), contrast-enhancement pattern (nonlayered/layered) and degree of enhancement (high/moderate/low). RESULTS: The most common type of gross appearance in both carcinomas was fungating, and the more common contrast-enhancement pattern in both carcinomas was a nonlayered pattern. The predominantly thickened layer was a high attenuation inner layer in both carcinomas. High-degree contrast enhancement was more common in SRC (37.1% of patients) than NSRC (15.6% of patients) with statistically significant difference (P = 0.01). CONCLUSIONS: Multidetector CT cannot distinguish SRC from NSRC based on the thickened stomach wall alone. But, high-degree contrast enhancement was more common in advanced gastric carcinoma with SRC than that with NSRC. PMID- 17082690 TI - Invasive carcinomas derived from intraductal papillary mucinous neoplasms of the pancreas: a long-term follow-up assessment with CT imaging. AB - OBJECTIVE: To describe computed tomography (CT) findings of invasive carcinoma derived from intraductal papillary mucinous neoplasms (IPMNs) of the pancreas during long-term follow-up. METHODS: Follow-up CT findings of 5 patients with IPMNs progressing to invasive carcinomas were respectively reviewed for 12 to 63 months. All patients underwent thin-section 3-phase helical and/or multislice CT. RESULTS: Invasive carcinomas were detected as hypo- (n = 3) or hyperattenuating (n = 2) solid masses in the pancreatic parenchyma on contrast-enhanced CT. Hypoattenuating masses were mostly visualized on arterial dominant phase images. In 4 branch-duct type IPMNs, the solid masses appeared with (n = 3) or without (n = 1) dilatation of the main pancreatic duct after 3 to 5 years. In the remaining combined-type IPMN, a solid mass was detected on initial CT and progressively increased during the follow-up. CONCLUSIONS: Arterial dominant phase CT is useful for detecting invasive carcinoma derived from IPMNs and is an effective follow-up method. PMID- 17082691 TI - MRI-based PSA density and PSA density of the transitional zone compared with PSA alone: correlation with prostate cancer Gleason score. AB - OBJECTIVES: The tumor Gleason score is an important prognostic factor in prostate cancer (PCA). This retrospective study analyzes whether serum prostate-specific antigen (PSA), or magnetic resonance imaging (MRI)-based PSA density of the entire prostate (PSAD) or the prostatic transitional zone (PSAT) distinguishes between PCA of Gleason scores 6 or lower (G6-) and 7 or higher (G7+). MATERIALS AND METHODS: Total prostate and transitional zone volumes were planimetrically determined in axial, T2-weighted fast spin echo (FSE) MRI images of the prostate in 61 patients with previously untreated PCA. Automated standardized microparticle enzyme immuno-assay (EIAs) measured PSA. RESULTS: Thirty patients had G6- and 31 patients had G7+. PSA values ranged from 1.0 to 57.2 ng/mL. Assignment to G6- or G7+, respectively, was correct in 49 of 61 (80%) cases (odds ratio [OR], 17.1; 95% confidence interval [CI], 4.8-61.5) for PSA above the optimal cutoff level of 10.35 ng/mL, 48 cases (79%; OR, 13.7; 95% CI, 4.0-46.8) for PSAD above the optimal cutoff level of 0.23 ng/mL/cm, and 45 cases (74%; OR, 6.9; 95% CI, 2.2-21.3) for PSAT above the optimal cutoff level of 0.38 ng/mL/cm (no significant differences, McNemar test). CONCLUSIONS: In patients with biopsy proven PCA, serum PSA level alone and MRI-based PSAD and PSAT help distinguish between G6- and G7+. PSAD and PSAT do not improve the level of confidence at which this discrimination is made. PMID- 17082692 TI - Magnetic resonance imaging for local staging of gastric carcinoma: results of an in vitro study. AB - OBJECTIVE: Preoperative staging of gastric carcinoma is limited by the fact that available imaging modalities do not enable accurate evaluation of the depth of infiltration of the gastric wall. The aim of this study was to evaluate the efficiency of conventional magnetic resonance imaging (MRI) in local staging of gastric carcinoma. METHODS: Sixty-five specimens of patients with proven gastric carcinoma were examined immediately after gastrectomy. Examination was performed with a 1-T MRI and included T1-weighted, T2-weighted, and opposed phase images. Images were analyzed for the number of visible wall layers and their signal intensity characteristics, for tumor localization and depth of infiltration. T stage was classified according to the TNM system. Finally, the staging by MRI was compared with the histopathological staging of the specimens. RESULTS: The mucosal, submucosal, and proper muscle layers could be differentiated by the typical signal intensities. Depiction of the subserosa or serosa was not possible. In 65 specimens, 67 carcinomas were found by the pathologist. Sixty four of 67 (96%) histologically proven carcinomas were correctly localized by MRI; T-staging accuracy was 50% only, mainly because of overstaging pT2 tumors as T3. CONCLUSIONS: MRI enables differentiation of gastric wall layers and, therefore, technically allows the evaluation of the local tumor stage of gastric carcinomas. However, infiltration of the subserosal and serosal layer cannot be proved accurately. Overstaging pT2 tumors is one of the most predominant problems. Yet further technical developments in high-resolution imaging of the gastric wall may improve T-staging in the near future and overcome today's staging limitations. PMID- 17082693 TI - Malignant fibrous histiocytoma of the common hepatic duct: a case report. AB - We report the first case of malignant fibrous histiocytoma of the common hepatic duct with radiologic findings and review of the literature, which was misdiagnosed as a cholangiocarcinoma of the common hepatic duct. PMID- 17082694 TI - Magnetic resonance imaging features of epidermoid cyst of the ovaries: magnetic resonance and computed tomography findings. AB - Epidermoid cyst of the ovary is a very rare tumor. There have been only 21 case reports of it previously. We describe the computed tomography (CT) and magnetic resonance (MR) findings of 2 cases of ovarian epidermoid cyst. To our knowledge, this is the first report describing CT and MR findings of epidermoid cysts of the ovary, and ruptured epidermoid cyst of the ovary has never been reported. PMID- 17082695 TI - MR imaging findings of pure prostatic leiomyoma: a report of two cases. AB - The magnetic resonance imaging findings in 2 patients pathologically proven to have pure leiomyoma of the prostate are reported. PMID- 17082696 TI - High-resolution computed tomographic angiography/computed tomographic cholangiography image fusion of the hepatobiliary system. AB - The 3-dimensional image was excellent as a result of the fast scanning and narrow collimation using multidetector computed tomography (CT); and the parallel escalation in the capabilities of the workstation allow the use of CT angiography/CT cholangiography image fusion. We demonstrated the feasibility of fusion images of the hepatobiliary system. We believe that fusion images are extremely valuable for applications such as preoperative planning for hepatic and bile duct resection. PMID- 17082697 TI - Unusual adrenal incidentalomas: magnetic resonance imaging features with pathological correlation. AB - Diagnosis of incidental adrenal masses is a real challenge to radiologists. The most common incidental tumors of the adrenal gland are adenomas and metastases. This article presents our experience with uncommon adrenal incidentalomas. Most of the magnetic resonance imaging characteristic features are demonstrated. When possible, they are correlated with the findings at gross and microscopic analysis, to help in understanding the mechanisms by which magnetic resonance imaging may aid in the characterization of the incidental adrenal mass. PMID- 17082698 TI - Multidetector CT angiography in pulmonary sequestration. AB - PURPOSE: To evaluate the role of multidetector CT angiography in the diagnosis and preoperative assessment of pulmonary sequestration with angiographic or surgical correlation. METHODS: Eight patients (5 males, 3 females) in the age range of 2 days to 35 years suspected of having pulmonary sequestration on the basis of clinical history and chest radiographs were included in the study. All patients underwent CT angiography (4 or 16 slice) and MPVR, MIP and 3D volume rendered images were generated. The axial images were also reviewed at soft tissue and lung window settings to evaluate the parenchymal changes. RESULTS: CT angiography showed five pulmonary sequestrations on the left and three on the right, located in the basal segments (n = 7) or paravertebral region (n = 1). Aberrant systemic arterial supply was demonstrated in all cases: from the descending thoracic aorta (n = 2); abdominal aorta (n = 3) and celiac axis (n = 3). Venous drainage into inferior pulmonary veins was demonstrated in 4 patients. The angioarchitecture depicted on CT angiography was confirmed on surgery in five patients and by DSA in two patients who subsequently underwent embolization using PVA particles and/or coils. CONCLUSION: The ability of CT angiography to simultaneously image the arterial supply, venous drainage and parenchymal changes in a single examination makes it the imaging modality of choice for the diagnosis and preoperative assessment of pulmonary sequestration. PMID- 17082699 TI - 3-dimensional adaptive raw-data filter: evaluation in low dose chest multidetector-row computed tomography. AB - OBJECTIVES: To evaluate a 3-dimensional adaptive raw-data filter in reducing streak artifacts in low dose chest computed tomographic (CT) images. METHODS: Fourteen adult patients who underwent low dose chest CT examination (parameters: 25 or 50 mAs, 120 kV) on 64-detector CTscanner were included in this study. We prepared 2 sets of contiguous 5-mm thick images by reconstruction with and without 3-dimensional adaptive raw-data filter (filter-processed and unprocessed images). Streak artifacts and visualization of peripheral vessels in both filter processed and unprocessed images were evaluated using a 5-point scale. Upper, middle, and lower thorax were evaluated separately. RESULTS: The difference in artifact severity was statistically significant in upper and lower thorax (P = 0.002 and 0.03, respectively), whereas it was not significant in middle thorax (P = 0.13). The difference in the visibility of peripheral pulmonary vessels was not statistically significant in all anatomical regions. CONCLUSIONS: The 3 dimensional adaptive raw-data filter reduced streak artifacts in low dose chest CT in upper and lower thorax. PMID- 17082700 TI - Remarkable anatomic anomalies of coronary arteries and their clinical importance: a multidetector computed tomography angiographic study. AB - OBJECTIVE: An anomalous origin and course of the coronary arteries can be benign or life threatening. Recently, because of new advances in computed tomography technology, radiologists have begun to interpret the diseases of coronary arteries. We aimed to demonstrate some remarkable anomalies of coronary arteries, some of which were not shown by multidetector computed tomography (MDCT) coronary angiography previously, and to discuss the clinical importance of these anomalies. MATERIALS AND METHODS: Seven hundred twenty-five consequent patients referred to Florence Nightingale Hospital and Ataturk University Hospital for MDCT coronary angiography were included in this study. The patients were between the ages of 33 and 78 years (mean +/- SD, 59 +/- 13.86 years). Four hundred ninety-seven patients (68.6%) were men, and 228 (31.4%) were women. All the examinations were evaluated by both a radiologist and a cardiologist. RESULTS: The incidence of anomalous anatomical origin and course of the coronaries found in our study group was 5.79% (n = 42). The anomalies found in our study are absence of the right coronary artery (RCA; n = 1, 0.13%), ectopic origin of RCA from the left anterior descending (LAD) artery (n = 1, 0.13%), absence of the left main coronary artery (n = 4, 0.52%), ectopic origin of the left main coronary artery from the right sinus of Valsalva (n = 1, 0.13%), double LAD and ectopic origin of LAD from RCA (n = 1, 0.13%), ectopic origin of the left circumflex artery from the right sinus of Valsalva (n = 3, 0.39%), ectopic origin of the left circumflex artery from RCA (n = 2, 0.26%), and myocardial bridging (n = 29, 4%). CONCLUSIONS: An anomalous origin of the coronary anatomy must be present in the interpretations because of its importance for patients, cardiologists, and surgeons. As a conclusion, our study showed that MDCT, especially volume rendering and maximum intensity projection techniques, may be useful for assessment of complex variations, when the conventional angiography may not be sufficient. PMID- 17082701 TI - Postmortem computed tomography of cadavers embalmed for use in teaching gross anatomy. AB - OBJECTIVE: To integrate radiology more fully into the first year of medical school, each student was provided with a computed tomographic (CT) scan of his or her gross anatomy cadaver before dissection. We describe the imaging findings of these embalmed cadavers. METHODS: Eighteen cadavers, embalmed within 2 days of death and stored an average of 8 months, were scanned with multidetector CT. RESULTS: The CT findings provided opportunities for enhanced learning and guidance for the dissection. Specific findings were categorized as postmortem and embalming changes, pathological changes, and iatrogenic changes. Image quality was degraded by beam hardening artifact caused by the arms being placed alongside the body. Differences in attenuation between many soft-tissue structures were reduced, compared with living subjects. CONCLUSIONS: Findings on CT scans of embalmed cadavers show significant differences from CT scans of living patients, due to postmortem state and artifacts of preservation. PMID- 17082702 TI - Computer-simulation technique for low dose computed tomographic screening. AB - OBJECTIVES: The purpose of this study is to assess the relative influence of noise and artifact in detecting lung nodules on low dose computed tomographic (CT) screening. METHODS: We develop the computer-simulation technique that allows tube current simulation and virtual nodule insertion in any CT images. The tube current simulation uses a reduction model that adds random Gaussian noise distribution to existing projection data. The virtual nodules are generated using a dedicated CT simulation tool with same scanner geometry. RESULTS: The coefficient of the correlations between the contrast-to-noise ratio of the actual scan and simulated tube current images was 0.98. There was no difference in CT number between virtual nodules and actual nodules [t test results = 0.60, t50(0.01) = 2.70 at 10 mA] and the coefficient of the correlations of the image noise was 0.99. CONCLUSIONS: Our technique is useful for systematic evaluation of radiation dose reduction and structure visibility in low-dose CT screening. PMID- 17082703 TI - Inherently 3-dimensional method for measurement of computed tomographic resolution anisotropy. AB - OBJECTIVE: Current techniques to measure computed tomography (CT) spatial resolution use separate methods for in-plane and out-of-plane directions. The growing use of near-isotropic voxel size necessitates a new single method that inherently measures resolution in any direction. METHOD: We introduce a method using a set of numerous glass microspheres suspended in a small volume from which a mean sphere image is constructed. Projecting asymptotes after imaging different microsphere sets with decreasing diameters provides an inherently 3-dimensional measure of spatial resolution and anisotropy. We apply the method to both a flat panel and multidetector CT scanner. RESULTS: The full-width at half-maximum from line profiles through mean sphere in transverse directions corresponds to known microsphere diameters. Increased longitudinal full-width at half-maximum corresponds to known anisotropy, which is larger for a multidetector CT scanner than for a flat-panel CT scanner. CONCLUSIONS: A new single method to measure CT resolution is inherently isotropic. PMID- 17082704 TI - Delayed MDCT imaging results in increased enhancement in patients with head and neck neoplasms. AB - OBJECTIVE: Using multidetector computed tomography, we compared the utility of a spilt bolus technique and the impact of performing delayed imaging compared with immediate imaging on tumor enhancement. METHODS: Thirty-seven patients with masses of the extracranial head and neck were imaged. An initial contrast bolus (50 cm) was administered over 30 seconds (1.67 cm/s) and an initial (immediate) imaging acquisition followed. Ninety seconds after completion of the initial bolus, a second bolus of 75 cm (2.5 cm/s) was administered and a second (delayed) acquisition obtained concurrently. Mean tumor enhancement in immediate and delayed imaging was calculated and analyzed using the paired t test. RESULTS: Mean enhancement was 68.3 +/- 21.0 Hounsfield units (HU) and 91.4 +/- 27.4 HU (n = 37) for immediate and delayed imaging groups, respectively. The mean difference between the initial and delayed enhancement was 23.1 HU (P < 0.01). CONCLUSIONS: The delayed imaging technique used resulted in a statistically significant increase in enhancement relative to immediate imaging. PMID- 17082705 TI - Magnetic resonance imaging characteristics with pathological correlation of cavernous malformation in cavernous sinus. AB - OBJECTIVE: To analyze extracerebral cavernous malformation located in the cavernous sinus and correlate their magnetic resonance imaging (MRI) features to pathological findings. MATERIALS AND METHODS: Nineteen patients (5 men and 14 women; mean age, 50.6 years; range, 32-71 years) with surgically verified cavernous malformations in the cavernous sinus were reviewed. MRI including T1 weighted, T2-weighted, and postcontrasted T1-weighted imaging was carried out in all instances with a 1.5-T superconductive system (Signa; General Electric Medical Systems, Milwaukee, WI). RESULTS: All these lesions measured from 2.5 cm to 6 cm (average, 4.3 cm) with 13 lesions located to the right side and 6 to the left side. Magnetic resonance images showed that 18 cases were hypointense to white matter on T1-weighted images with only one case showed mixed hypointensity and hyperintensity. On T2-weighted images, 17 cases showed marked homogeneous hyperintensity, 1 case showed marked hyperintensity with some signal void structures and 1 with mixed intensity. Marked homogeneous enhancement after contrast material administration was found in 7 cases, and the remaining 12 showed marked heterogeneous enhancement. Pathologically, these lesions can be classified as type A, type B, and type C. Type A was sponge-like with intact pseudocapsule; type B was mulberry-like with the pseudocapsule incomplete or absent; and type C was composed of both mulberry-like composition and sponge-like composition. Lesions with homogeneous contrast enhancement on MRI correlated with type A pathological findings, whereas those with heterogeneous enhancement correlated with type B and type C pathological findings. Asymmetrical dumbbell shaped masses of 13 cases involving the sellar and round masses of 6 cases with 2 protruding into the sellar were found. All of the cavernous malformations displaced the adjacent temporal lobe without adjacent brain edema. The internal carotid arteries were displaced or encased in all cases. CONCLUSIONS: If a well demarcated, homogeneous high signal intensity lesion on T2-weighted images with a dumbbell configuration involving both parasellar and sellar regions with marked heterogeneous or homogeneous enhancement is found, the diagnosis of cavernous sinus cavernous malformation should be entertained. PMID- 17082706 TI - Capillary telangiectasia of the brain stem diagnosed by susceptibility-weighted imaging. AB - A case of brain stem capillary telangiectasia diagnosed by susceptibility weighted imaging is reported. A small enhancing pontine lesion was found on postcontrast T1-weighted MR images in a 56-year-old woman with human T-cell leukemia virus type 1 infection. Imaging diagnosis was difficult with conventional MR imaging because the lesion did not show characteristic signal loss on conventional gradient-echo images. SWI was useful for imaging diagnosis as it demonstrated marked signal loss of the lesion. PMID- 17082707 TI - Measurement of tumor blood flow using dynamic contrast-enhanced magnetic resonance imaging and deconvolution analysis: a preliminary study in musculoskeletal tumors. AB - OBJECTIVE: To measure tumor blood flow (TBF) using dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI). METHODS: A DCE-MRI was performed using inversion recovery-preparation fast-field echo sequences. Dynamic data were obtained every 3.2 seconds for 2 minutes, immediately after gadolinium injection. In 14 patients with malignant musculoskeletal tumors, TBF maps were generated pixel-by-pixel by deconvolution analysis. For preclinical studies, muscle blood flow in 5 volunteers and signal intensities of different gadolinium concentrations were measured. RESULTS: There was a good linear relationship between signal intensities and gadolinium concentrations (r = 0.989, P < 0.001, at gadolinium concentrations or = 140 and/or 90 mmHg and a SBPM or daytime ABPM value < 135/85 mmHg. This definition was used irrespective of treatment. We also determined the prevalence of a substantial WCE (OBPM 20 mmHg systolic or 10 mmHg diastolic higher than SBPM or daytime ABPM). SETTING: Patients were recruited at hospital or general practice. PATIENTS: A total of 163 mild-to-moderate essential hypertensive patients with a mean age of 56 years (56% males). RESULTS: At eight blood pressure (BP) measurement occasions, 75% of all patients had a substantial WCE at least once, while 57% had WCH at least once. One-third of the patients consistently had a substantial WCE and 14% consistently had WCH on three or more occasions The magnitude of the WCE was significantly related to the height of blood pressure in treated but not in untreated patients. CONCLUSION: In some patients, WCH or a substantial WCE occurs consistently on multiple OBPM visits. Especially in untreated patients, the magnitude of the WCE varies widely among individuals. These results support the incorporation of SBPM and/or ABPM into optimal management of hypertension, not only to prevent misdiagnosis in untreated patients but also to determine the need for adjusting antihypertensive therapy in treated subjects. PMID- 17082717 TI - Predictive factors for masked hypertension within a population of controlled hypertensives. AB - CONTEXT: Prevalence of masked hypertension (MH) is far from negligible reaching 40% in some studies. The SHEAF study (Self measurement of blood pressure at Home in the Elderly: Assessment and Follow-Up) and others clearly showed that masked hypertension (MH) as detected by home blood pressure measurement (HBPM) is associated with poor cardiovascular prognosis. OBJECTIVE: Systematic HBPM to detect MH is not yet routine. The aim of this work is to better define the clinical profile of masked hypertensives within a population with controlled office blood pressure (BP) and the factors associated with a higher prevalence of MH. MATERIALS AND METHODS: BP was measured at the clinic by the doctor and at home by the patient himself. Risk factors for MH were analysed in a cohort of 1150 treated hypertensive patients over the age of 60 (mean age 70 +/- 6.5, 48.9% men) with controlled office BP. (SBP < 140 mmHg and DBP < 90 mmHg). RESULTS: 463 patients (40%) were masked hypertensives (SBP > or = 135 mmHg or DBP > or = 85 mmHg at home). Three parameters were associated with MH (odds ratio OR): office SBP (OR = 1.110), male gender (OR = 2.214) and age (OR = 1.031). Decision trees showed a 130 mmHg SBP was an efficient threshold to propose HBPM with a higher probability to detect MH. Subsequent variables were male gender and age over 70 in males. CONCLUSION: To detect masked hypertension, it would be logical to first of all select patients whose office SBP is between 130 and 140 mmHg. PMID- 17082718 TI - Aortic stiffness is associated with atherosclerosis of the coronary arteries in older adults: the Rotterdam Study. AB - OBJECTIVE: Aortic stiffness can lead to low diastolic blood pressure, thereby possibly limiting coronary perfusion. Therefore, the simultaneous occurrence of both aortic stiffness and coronary atherosclerosis can lead to an increased risk of subendocardial ischaemia. The aim of the present study was to investigate the association between aortic stiffness and coronary atherosclerosis. METHODS: The study was performed in 1757 subjects of the Rotterdam Study, a population-based study of elderly individuals. Aortic stiffness was assessed by measuring carotid femoral pulse wave velocity (PWV). Coronary atherosclerosis was assessed by measuring coronary calcification using electron beam tomography and expressed as a total calcium score. The total calcium score was log-transformed because of its skewed distribution. The association between PWV and coronary calcification was first evaluated after adjustment for age, sex, mean arterial blood pressure and heart rate. RESULTS: Linear regression analyses showed that increased PWV was associated with a higher log total coronary calcium score [beta-regression coefficient 0.11, 95% confidence interval (CI) 0.07-0.15]. Compared with the lowest quartile of PWV, multivariate odds ratios and corresponding 95% CI for advanced coronary calcification in the second, third and fourth highest quartiles were 1.17 (0.79-1.74), 1.58 (1.07-2.34) and 2.12 (1.40-3.20), respectively. CONCLUSIONS: In this large population-based study performed in elderly subjects aortic stiffness was strongly and independently associated with coronary atherosclerosis. PMID- 17082719 TI - High pulse pressure and low mean arterial pressure: two predictors of death after a myocardial infarction. AB - OBJECTIVES: Although the negative prognostic implication of a clinical history of arterial hypertension in myocardial infarction (MI) survivors is well known, the predictive role of the blood pressure (BP) regimen after MI is not well defined. The aim of this study was to investigate the prognostic significance of different BP indices in post-MI. METHODS AND RESULTS: We evaluated the relationship between baseline systolic, diastolic, pulse and mean arterial pressure (MAP), measured by sphygmomanometry at discharge from hospital or within 3 months of an MI, and total and cardiovascular mortality in 11 116 patients enrolled in the GISSI Prevenzione trial. Over 3.5 years of follow-up, 999 patients died, 657 of them from cardiovascular causes. Low mean and high pulse pressure were significantly associated with total and cardiovascular mortality after controlling for potential confounders in the multivariate analysis. As compared with patients with less extreme BP values, patients with MAP of 80 mmHg or less (n = 1241; 11.2%) had a 48% higher risk of cardiovascular death [95% confidenceinterval (CI) 1.16-1.87; P = 0.001] and those with pulse pressure greater than 60 mmHg (n = 958; 8.6%) had a 35% higher risk (95% CI 1.09-1.69; P = 0.007); only four subjects (0.04%) had both a high pulse pressure and a low MAP (relative risk of cardiovascular death 3.48; 95% CI 0.48-25.88; P = 0.218). CONCLUSIONS: Our results show for the first time an additional prognostic importance of two easily measurable components of BP, definitely high pulse pressure (> 60 mmHg) and low MAP (< or = 80 mmHg), in a large sample of non-selected patients surviving MI who entered a modern programme of cardiovascular prevention. PMID- 17082720 TI - Female sexual dysfunction in essential hypertension: a common problem being uncovered. AB - OBJECTIVES: Female sexual dysfunction (FSD) is increasingly attracting more scientific and public interest, and represents a poorly investigated issue in patients with essential hypertension. We evaluated the prevalence of sexual dysfunction in hypertensive women compared with normotensive women according to age, hypertension severity, hypertension duration, and antihypertensive treatment. METHODS: The study population consisted of consecutive, sexually active women attending an outpatient hypertension clinic. The Female Sexual Function Index (FSFI questionnaire) was used to evaluate FSD. Univariate and multivariate analyses were used to evaluate predictors of FSD. RESULTS: Four hundred and seventeen women were studied. From them, 216 women had arterial hypertension (136 treated, 80 untreated) and 201 were normotensive. Sexual dysfunction was found in 42.1% of hypertensive women compared with 19.4% of normotensive women (odds ratio, 3.2; 95% confidence interval, 1.9-4.7; P < 0.001). Systolic blood pressure levels were significantly related to FSFI score (r = -0.67, P < 0.001). Successful control of hypertension was related to lower prevalence of FSD. Increasing age (beta = -0.187, P = 0.001), increasing systolic blood pressure (beta = -0.687, P < 0.001), and beta-blocker administration (beta = -0.162, P = 0.001) were significant predictors of sexual dysfunction in this patient population. CONCLUSIONS: FSD is more prevalent in women with essential hypertension compared with women with normal blood pressure, and its prevalence declines with adequate blood pressure control. Adequate control of hypertension with medication not affecting sexual function can have a great impact on the quality of life of hypertensive patients. Physicians should recognize and properly manage FSD in hypertensive women. PMID- 17082721 TI - Endothelial nitric oxide synthase haplotypes are related to blood pressure elevation, but not to resistance to antihypertensive drug therapy. AB - OBJECTIVES: Most hypertensive patients require two or more drugs to control arterial blood pressure effectively. Although endothelial nitric oxide synthase (eNOS) haplotypes have been associated with hypertension, it is unknown whether eNOS genotypes/haplotypes are associated with resistance to antihypertensive therapy. METHODS: We studied the distribution of three eNOS genetic polymorphisms: single nucleotide polymorphisms in the promoter region (T(-786)C), and in exon 7 (Glu298Asp), and a variable number of tandem repeats in intron 4 (b/a). Genotypes were determined for 111 normotensive controls (NT), 116 hypertensive individuals who were well controlled (HT), and 100 hypertensive individuals who were resistant to conventional antihypertensive therapy (RHT). We also compared the distribution of eNOS haplotypes in the three groups of subjects. RESULTS: No differences were found in genotype or allele distribution among the three groups (all P > 0.05). Conversely, the 'C Glu b' haplotype was more commonly found in the NT than in the HT or RHT groups (21 versus 8 and 7%, respectively; both P < 0.00625). In addition, the 'C Asp b' haplotype was more commonly found in the HT or RHT groups than in the NT group (22 and 20%, respectively, versus 8%; both P < 0.00625). The distribution of eNOS haplotypes was not significantly different in the HT and RHT groups (P > 0.05). CONCLUSIONS: Whereas our findings suggest a protective effect for the 'C Glu b' haplotype against hypertension and that the 'C Asp b' haplotype increases the susceptibility to hypertension, our results suggest that eNOS haplotypes are not associated with resistance to antihypertensive therapy. PMID- 17082722 TI - Impaired vasodilation in response to perivascular nerve stimulation in mesenteric arteries of TRPV1-null mutant mice. AB - BACKGROUND: The role of the transient receptor potential vanilloid type 1 (TRPV1) channels expressed in perivascular sensory nerves in the regulation of vascular reactivity is largely unknown. This study was designed to test the hypothesis that vasodilation induced by electrical field stimulation (EFS) of perivascular sensory nerves is mediated by the TRPV1 via release of sensory neurotransmitters in wild-type (WT) mice, and this effect is abolished in gene-targeted TRPV1-null mutant (TRPV1(-/-)) mice. METHODS: Isolated mesenteric resistance arteries from WT and TRPV1(-/-)) mice were perfused and pretreated with guanethedine and atropine to block sympathetic and parasympathetic nerve activity, respectively. After precontracting with phenylephrine, changes of vascular diameters induced by EFS were monitored in the absence or presence of the TRPV1 receptor antagonist capsazepine; the calcitonin gene-related peptide (CGRP) receptor antagonist, CGRP8-37; or the substance P (SP) receptor antagonist, RP67580. RESULTS: EFS induced vasodilation was significantly reduced in arteries in TRPV1(-/-)) mice when compared to that of WT mice. Capsazepine and CGRP8-37 attenuated vasodilation induced by EFS in WT but not TRPV1(-/-)) mice. In contrast, RP67580 had no effect on the EFS-induced vasodilation in WT or TRPV1(-/-)) mice. The release of CGRP in the face of EFS challenge was significantly increased in both WT and TRPV1(-/-)) arteries, which was attenuated by capsazepine in WT but not TRPV1(-/-)) arteries. Exogenous CGRP caused dose-dependent vasodilation to a similar degree in WT and TRPV1(-/-)) arteries. CONCLUSIONS: Our data show that in WT mice transmural stimulation of perivascular sensory nerves activates the TRPV1, leading to CGRP release from sensory nerve endings; and blockade of CGRP, but not SP, receptors abolishes TRPV1-mediated vasodilation during EFS. All these effects are impaired in TRPV1(-/-)) mice, indicating that TRPV1 plays a key role in modulating perivascular sensory nerve-mediated vasodilation. PMID- 17082723 TI - The role of stress hormones in the relationship between resting blood pressure and coagulation activity. AB - BACKGROUND: Systemic hypertension confers a hypercoagulable state. We hypothesized that resting mean blood pressure (MBP) interacts with stress hormones in predicting coagulation activity at rest and with acute mental stress. METHODS: We measured plasma clotting factor VII activity (FVII:C), FVIII:C, fibrinogen, D-dimer, epinephrine and norepinephrine, and saliva cortisol in 42 otherwise healthy normotensive and hypertensive medication-free men (mean age 43 +/- 14 years) at rest, immediately after stress, and twice during 60 min of recovery from stress. RESULTS: At rest, the MBP-by-epinephrine interaction predicted FVII:C (beta = -0.33, P < 0.04) and D-dimer (beta = 0.26, P < 0.05), and the MBP-by-cortisol interaction predicted D-dimer (beta = 0.43, P = 0.001), all independent of age and body mass index (BMI). Resting norepinephrine predicted fibrinogen (beta = 0.42, P < 0.01) and D-dimer (beta = 0.37, P < 0.03), both independent of MBP. MBP predicted FVIII:C change from rest to immediately post-stress independent of epinephrine (beta = -0.37, P < 0.03) and norepinephrine (beta = -0.38, P < 0.02). Cortisol change predicted FVIII:C change (beta = -0.30, P < 0.05) independent of age, BMI and MBP. Integrated norepinephrine change from rest to recovery (area under the curve, AUC) predicted D-dimer AUC (beta = 0.34, P = 0.04) independent of MBP. The MBP-by-epinephrine AUC interaction predicted FVII:C AUC (beta = 0.28) and fibrinogen AUC (beta = 0.30), and the MBP-by-norepinephrine AUC interaction predicted FVIII:C AUC (beta = -0.28), all with borderline significance (Ps < 0.09) and independent of age and BMI. CONCLUSIONS: MBP significantly altered the association between stress hormones and coagulation activity at rest and, with borderline significance, across the entire stress and recovery interval. Independent of MBP, catecholamines were associated with procoagulant effects and cortisol reactivity dampened the acute procoagulant stress response. PMID- 17082724 TI - Angiotensin II/AT2 receptor-induced vasodilation in stroke-prone spontaneously hypertensive rats involves nitric oxide and cGMP-dependent protein kinase. AB - BACKGROUND: Angiotensin II (Ang II) induces vasodilation, in part, through angiotensin type 2 receptor (AT2R)-induced actions in conditions associated with angiotensin type 1 receptor (AT1R) blockade and AT2R upregulation. Ang II/AT2R induced vasodilation involves nitric oxide (NO)-cyclic guanosine monophosphate (cGMP)-dependent processes. We previously demonstrated that AT2R-mediated effects involve inhibition of the RhoA/Rho kinase pathway. However, molecular mechanisms underlying this phenomenon are unknown. AIMS: In the present in-vivo study we tested the hypothesis that AT2R-elicited vasodilation is associated with nitric oxide synthase (NOS) activation and NO production, and that a cGMP-dependent protein kinase (cGKI), which inactivates RhoA, is upregulated when stroke-prone spontaneously hypertensive rats (SHRSP) are treated with AT1R blockers. METHODS: SHRSP and Wistar-Kyoto (WKY) rats were treated with the AT1R blocker valsartan for 14 days. Dilatory responses to Ang II with or without the NOS inhibitor N nitro-L-arginine methyl ester (L-NAME) were performed in norepinephrine precontracted vessels in the presence of valsartan. Expression of AT2R, endothelial NOS (eNOS) and cGKI was assessed by immunoblotting. NO bioavailability and NAD(P)H oxidase activity were evaluated by chemiluminescence. RESULTS: Ang II elicited vasodilation in valsartan-treated SHRSP. L-NAME inhibited this effect, indicating a role for NO. eNOS expression and NO concentration were increased twofold by valsartan, only in SHRSP. Expression of cGKI was reduced in SHRSP and restored after valsartan treatment. NAD(P)H oxidase activity was approximately threefold higher in SHRSP versus WKY (P < 0.05) and reduced by valsartan. CONCLUSIONS: Ang II, via AT2R, facilitates vasodilation through NOS/NO-mediated pathways and upregulation [corrected] of CGK1 [corrected] after chronic AT1R antagonism. These effects may contribute in part to beneficial actions of AT1R blockers in the treatment of hypertension. PMID- 17082725 TI - Simvastatin treatment modifies polymorphonuclear leukocyte function in high-risk individuals: a longitudinal study. AB - BACKGROUND: Although extensive experimental evidence supports a primary role of polymorphonuclear leukocytes (PMNs) in atherosclerosis, few data exist concerning the functional properties of these cells and their pharmacological modulation in high-risk individuals. OBJECTIVE: The production of the proinflammatory chemokine interleukin-8 (IL-8), migration and chemotaxis, and reactive oxygen species (ROS) generation were investigated in a longitudinal study in PMNs obtained from high risk individuals during statin treatment. As a secondary endpoint we compared PMN function of high-risk patients with that of controls. METHODS AND RESULTS: PMNs were isolated from 21 high-risk individuals before treatment and 3 and 30 days after the beginning of simvastatin treatment, and from healthy controls. During treatment a significant reduction was observed both in resting (P = 0.009) and N formyl-Met-Leu-Phe (fMLP)-stimulated (P = 0.008) IL-8 production, and in the chemotactic index (P = 0.038), whereas ROS generation did not significantly change. In comparison with cells from controls, PMNs obtained from patients before starting simvastatin treatment showed higher resting and fMLP-stimulated IL-8 release (P = 0.007 and P = 0.002, respectively) and ROS generation (resting, P = 0.009; and fMLP-stimulated, P = 0.046), whereas migration and the chemotactic index did not significantly differ. CONCLUSIONS: An activation of neutrophils is present in high-risk individuals, shown by the enhanced production of IL-8, and increased ROS generation. The 4-week statin treatment is able to reduce the cell capability to produce IL-8, and to decrease chemotaxis, thus affecting the proinflammatory properties of PMNs. PMID- 17082726 TI - Ambulatory blood pressure monitoring in obese children: role of insulin resistance. AB - OBJECTIVES: To investigate the relationship between ambulatory blood pressure (ABPM) parameters and insulin resistance in obese children. METHODS: A population of 56 obese prepubertal children was recruited for the study. They underwent ABPM, an oral glucose tolerance test and complete physical examination, including adiposity indexes such as body mass index (BMI), skinfolds, waist-to-hip ratio (WHR) and fat mass. RESULTS: The standard deviation score for BMI was significantly correlated with 24-h systolic blood pressure (SBP) (r = 0.30; P = 0.02) and diastolic blood pressure (DBP) (r = 0.29; P = 0.03), daytime SBP and DBP (r = 0.28; P = 0.04 and r = 0.32; P = 0.02), night-time SBP and DBP (r = 0.32; P = 0.01 and r = 0.27; P = 0.04). Fat mass was correlated with 24-h SBP (r = 0.46; P = 0.005), daytime SBP (r = 0.40; P = 0.01) and night-time SBP (r = 0.49; P = 0.03). No correlations were found between ABPM parameters and WHR. Furthermore, significant correlations were found between insulin resistance indexes, such as the homeostasis model assessment of insulin resistance and quantitative insulin-sensitivity check index, and 24-h DBP (r = 0.34; P = 0.01 and r = -0.29; P = 0.03), daytime DBP (r = 0.35; P = 0.009 and r = -0.34; P = 0.01) and daytime SBP (r = 0.32; P = 0.02 and r = -0.27; P = 0.04). Only 24-h and daytime DBP remained correlated with insulin resistance after adjustment for obesity. The analysis of the circadian rhythm of blood pressure revealed that 24 out the 56 children were non-dippers. CONCLUSIONS: The results of the present study indicate that adiposity and insulin resistance have an important role in influencing blood pressure in obese children, and also show a high prevalence of non-dipping phenomenon. This is of particular relevance because blood pressure tracks from childhood into adulthood and an already early-life high blood pressure is associated with an increased cardiovascular risk. PMID- 17082727 TI - Baroreflex sensitivity improvement is associated with decreased oxidative stress in trained spontaneously hypertensive rat. AB - BACKGROUND: Baroreflex sensitivity (BRS) impairment has been associated with endothelial dysfunction and oxidative stress. METHODS: Because exercise training could improve endothelial function in spontaneously hypertensive rats (SHR), the effect of moderate exercise training on oxidative stress and BRS was investigated. Groups were divided into sedentary and trained Wistar-Kyoto rats (S WK, n = 7 and T-WK, n = 6) and SHR (S-SHR and T-SHR, n = 9 each). Exercise training was performed on a treadmill (5 days/week, 60 min, 10 weeks), and the lactate threshold (20 m/min) was used to determine moderate intensity. RESULTS: Exercise training reduced mean arterial pressure in WK and SHR (S-WK 127 +/- 4, T WK 105 +/- 5, S-SHR 169 +/- 4 versus T-SHR 140 +/- 4 mmHg; P < 0.01). Baroreflex bradycardic (S-WK -1.89 +/- 0.15, T-WK -2.11 +/- 0.37, S-SHR -0.80 +/- 0.09 versus T-SHR -1.29 +/- 0.10 bpm/mmHg; P < 0.0001) and tachycardic (S-WK 2.57 +/- 0.19, T-WK 2.73 +/- 0.21, S-SHR 1.18 +/- 0.07 versus T-SHR 2.02 +/- 0.10 bpm/mmHg; P < 0.0001) responses were significantly different between groups. Lipoperoxidation in erythrocytes (S-WK 11 320 +/- 739, T-WK 10 397 +/- 765, S-SHR 20 511 +/- 1627 versus T-SHR 10 211 +/- 589 counts per second (cps)/mg haemoglobin; P < 0.0001) and aortas (S-WK 12 424 +/- 2219, T-WK 7917 +/- 726, S SHR 26 957 +/- 1772 versus T-SHR 17 777 +/- 1923 cps/mg protein; P < 0.0001) was reduced in T-SHR compared with S-SHR. Inverse correlations were observed between both bradycardic and tachycardic responses and lipoperoxidation in erythrocytes (r = 0.56 and r = -0.77, respectively; P < 0.01) and aortas (r = 0.77 and r = 0.80, respectively; P < 0.0001). CONCLUSION: Our results indicate that exercise training decreases oxidative stress, which is related to an improvement in BRS in SHR. PMID- 17082728 TI - Pre-eclampsia is associated with Helicobacter pylori seropositivity in Italy. AB - OBJECTIVES: Pre-eclampsia (PE) is characterized by an excess of inflammation and endothelial dysfunction. Helicobacter pylori (H. pylori) causes chronic inflammatory changes and endothelial damage. We investigated the prevalence of seropositivity for IgG against H. pylori and cytotoxin-associated antigen A (CagA) in PE patients and the presence of H. pylori DNA in their placentas. METHODS: We tested 47 pregnant women with PE and 47 with uneventful pregnancies for serum antibodies against H. pylori (enzyme immunoassays) and CagA protein (immunoblot assays). In 20 of them (10 normal and 10 PE) we assessed the presence, in the placenta, of H. pylori DNA by means of nested polymerase chain reaction (PCR). The odds ratios (OR) and 95% confidence intervals (CI), adjusted for parity, were calculated using logistic regression analysis to assess the risk of PE associated with H. pylori infection. RESULTS: Helicobacter pylori seropositivity frequency was higher in mothers with PE (51.1%) compared to women with uneventful pregnancy (31.9%) (OR, 2.668; 95% CI, 1.084-6.566; P = 0.033). The difference was even greater for CagA seropositivity (80.9 and 14.9%, respectively) (OR, 26.035; 95% CI, 8.193-82.729; P < 0.001). All placentas were negative for H. pylori DNA. CONCLUSIONS: Helicobacter pylori, and especially strains carrying the CagA gene, may contribute to the inflammatory mechanisms involved in the pathogenesis of PE. PMID- 17082729 TI - Chronic nitric oxide blockade modulates renal Na-K-2Cl cotransporters. AB - OBJECTIVE: The Na-K-2Cl cotransporter (NKCC2 isoform) of the thick ascending limb of Henle's loop (TAL) plays an important role in renal sodium handling, and the vascular isoform (NKCC1) participates in the response to vasoconstrictors. Both isoforms appear to be regulated by nitric oxide. This study aimed to analyze the effect of chronic nitric oxide deficiency on tubular and vascular Na-K-2Cl cotransporters in kidney and their potential role in the development of N-nitro-L arginine-methyl ester (L-NAME) hypertension. METHODS: Wistar rats were given L NAME (vehicle, 10, 35 and 80 mg/100 ml drinking water) for 4 weeks. Blood pressure was measured by the tail-cuff method. NKCC2 activity was estimated as the bumetanide-sensitive Rb influx in fresh isolated TAL tubules. NKCC1 contractile function was estimated as the bumetanide-sensitive vasocontractile response to phenylephrine in isolated perfused kidneys. Acute effects of L-NAME and endothelium removal were also evaluated. NKCC2 and NKCC1 protein expression were assessed by western blot analysis. RESULTS: Chronic L-NAME administration increased, in a dose-dependent manner, both blood pressure and NKCC2 activity, and these changes significantly correlated (r2 = 0.89, P < 0.01). NKCC1 contractile activity decreased with the highest dose of L-NAME (80 mg/100 ml drinking water group) but it was not affected by acute nitric oxide blockade or endothelium removal. This 80 mg group showed increased NKCC2 expression in the renal medulla and decreased NKCC1 expression in aorta. CONCLUSIONS: Chronic nitric oxide deficiency stimulates tubular Na-K-2Cl cotransporter, suggesting that NKCC2 hyperactivity contributes to the inability to excrete sodium, and hence to the development of L-NAME hypertension. In contrast, L-NAME hypertension develops independently of vascular NKCC1-contractile activity. PMID- 17082730 TI - QT interval in patients with primary aldosteronism and low-renin essential hypertension. AB - INTRODUCTION: QT interval prolongation increases the risk of sudden death in several medical conditions. Patients with primary aldosteronism and salt sensitive hypertension experience more cardiovascular events than those with normal-renin essential hypertension. QT interval prolongation might represent one of the risk factors for cardiovascular events in these patients. The aim of the present study was to evaluate the QT interval in patients with primary aldosteronism and low-renin essential hypertension (LREH). METHODS: Twenty-seven patients with primary aldosteronism, 17 patients with LREH, 117 patients with essential hypertension and 25 healthy individuals were studied. Plasma aldosterone, plasma renin activity, and aldosterone to plasma renin activity ratio (ARR) were determined. Corrected QT intervals (QTcs) were measured from a 12-lead electrocardiogram. RESULTS: The QTc was longer in primary aldosteronism (434 +/- 23 ms) and LREH (430 +/- 18 ms) compared with essential hypertension (419 +/- 22 ms) and healthy controls (412 +/- 19 ms) (P = 0.0004). The prevalence of QTc longer than 440 ms was higher in primary aldosteronism (48%) and LREH (23%) compared with essential hypertension (11%) and healthy controls (4%) (P < 0.0001). QTc correlated with plasma aldosterone (P = 0.01), ARR (P = 0.02), and diastolic blood pressure (P = 0.01). ARR (P = 0.01) and systolic blood pressure (P = 0.01) were identified as independent predictors of QTc. CONCLUSIONS: We postulate that the elevated aldosterone secretion contributes to the prolongation of the QT interval in patients with primary aldosteronism and LREH through both a depletion of intracellular potassium concentration and higher blood pressure values. QTc measurement might represent one simple, non-invasive and reproducible index to characterize the cardiovascular risk in patients with primary aldosteronism and LREH. PMID- 17082731 TI - AT1 receptor blockade is superior to conventional triple therapy in protecting against end-organ damage in Cyp1a1-Ren-2 transgenic rats with inducible hypertension. AB - OBJECTIVE: In the present study we compared the effects of treatment with the AT1 receptor antagonist candesartan and of 'triple therapy' (hydralazine, hydrochlorothiazide, reserpine) on the course of blood pressure, cardiac hypertrophy and angiotensin II concentrations after induction of hypertension in transgenic rats with inducible expression of the mouse renin gene (Cyp1a1-Ren-2 rats). METHODS: Hypertension was induced in Cyp1a1-Ren-2 rats through dietary administration of the natural xenobiotic indole-3-carbinol (I3C, 0.3%) for 4 days. Starting on the day before administration of I3C, rats were treated either with candesartan or received triple therapy for 9 days. Systolic blood pressure was measured in conscious animals. Rats were decapitated and angiotensin II levels in plasma and in whole kidney and left ventricular tissues were determined by radioimmunoassay. RESULTS: Administration of I3C resulted in the development of severe hypertension and cardiac hypertrophy that was accompanied by marked elevations of plasma and tissue angiotensin II concentrations. Candesartan treatment prevented the development of hypertension and cardiac hypertrophy and was associated with a reduction of tissue angiotensin II concentrations. In contrast, triple therapy, despite maintaining systolic blood pressure in the normotensive range, did not prevent the development of cardiac hypertrophy and tissue angiotensin II augmentations. CONCLUSIONS: Our findings indicate that hypertension in Cyp1a1-Ren-2 rats is a clearly angiotensin II-dependent model of hypertension with elevated circulating and tissue angiotensin II concentrations, and that antihypertensive treatment with AT1 receptor blockade is superior to conventional triple therapy in effective protection against hypertension-induced end-organ damage in this rat model. PMID- 17082732 TI - Functional significance of single nucleotide polymorphisms within the 5'-flanking region of beta2-adrenergic receptor gene. PMID- 17082733 TI - Baseline and target blood pressure for the prevention of recurrent stroke. PMID- 17082735 TI - European Society of Hypertension Scientific Newsletter: control of hypertension in patients with peripheral artery disease. PMID- 17082736 TI - European Society of Hypertension Scientific Newsletter: update on hypertension management: prevention of type 2 diabetes mellitus with antihypertensive drugs. PMID- 17082737 TI - European Society of Hypertension Scientific Newsletter: treatment of hypertensive urgencies and emergencies. PMID- 17082738 TI - Treatment of Clostridium difficile-associated disease (CDAD). PMID- 17082739 TI - Decitabine (Dacogen) for myelodysplastic syndromes. PMID- 17082740 TI - Evidence of indigenous NAH plasmid of naphthalene degrading Pseudomonas putida PpG7 strain implicated in limonin degradation. AB - A well characterized naphthalene-degrading strain, Pseudomonas putida PpG7 was observed to utilize limonin, a highly-oxygenated triterpenoid compound as a sole source of carbon and energy. Limonin concentrations evidenced a 64% reduction over 48 h of growth in batch cultures. Attempts were made to acquire a plasmid less derivative via various methods (viz. Ethidium Bromide, SDS, elevated temperature & mitomycin C), among which the method involving mitomycin C (20 ug/ml) proved successful. Concomitant with the loss of plasmid in P. putida PpG7 strain, the cured derivative was identified as a lim- phenotype. The lim+ phenotype could be conjugally transferred to the cured derivative. Based on the results of curing with mitomycin C, conjugation studies and presence of ndo gene encoding naphthalene 1,2 dioxygenase, it was demonstrated that genes for the limonin utilization were encoded on an 83 kb indigenous transmissible Inc. P9 NAH plasmid in Pseudomonas putida PpG7 strain. PMID- 17082741 TI - Lichen flora around the Korean Antarctic Scientific Station, King George Island, Antarctic. AB - As part of the long-term monitoring projects on Antarctic terrestrial vegetation in relation to global climate change, a lichen floristical survey was conducted around the Korean Antarctic Station (King Sejong Station), which is located on Barton Peninsula, King George Island, in January and February of 2006. Two hundred and twenty-five lichen specimens were collected and sixty-two lichen species in 38 genera were identified by morphological characteristics, chemical constituents, TLC analysis and ITS nucleotide sequence analysis. PMID- 17082742 TI - Heat shock causes oxidative stress and induces a variety of cell rescue proteins in Saccharomyces cerevisiae KNU5377. AB - In this study, we attempted to characterize the physiological response to oxidative stress by heat shock in Saccharomyces cerevisiae KNU5377 (KNU5377) that ferments at a temperature of 40 degrees C. The KNU5377 strain evidenced a very similar growth rate at 40 degrees C as was recorded under normal conditions. Unlike the laboratory strains of S. cerevisiae, the cell viability of KNU5377 was affected slightly under 2 hours of heat stress conditions at 43 degrees C. KNU5377 evidenced a time-dependent increase in hydroperoxide levels, carbonyl contents, and malondialdehyde (MDA), which increased in the expression of a variety of cell rescue proteins containing Hsp104p, Ssap, Hsp30p, Sod1p, catalase, glutathione reductase, G6PDH, thioredoxin, thioredoxin peroxidase (Tsa1p), Adhp, Aldp, trehalose and glycogen at high temperature. Pma1/2p, Hsp90p and H+-ATPase expression levels were reduced as the result of exposure to heat shock. With regard to cellular fatty acid composition, levels of unsaturated fatty acids (USFAs) were increased significantly at high temperatures (43 degrees C), and this was particularly true of oleic acid (C18:1). The results of this study indicated that oxidative stress as the result of heat shock may induce a more profound stimulation of trehalose, antioxidant enzymes, and heat shock proteins, as well as an increase in the USFAs ratios. This might contribute to cellular protective functions for the maintenance of cellular homeostasis, and may also contribute to membrane fluidity. PMID- 17082743 TI - Effect of mycelial extract of Clavicorona pyxidata on acetylcholinesterase and beta-secretase activity in vitro. AB - In a previous study, an extract of Clavicorona pyxidata DGUM 29005 mycelia demonstrated an inhibitory effect against enzyme-associated perceptual disorders. We have attempted to determine whether this mycelial extract is also capable of inhibiting the activities of acetylcholinesterase (AChE) and beta-secretase (BACE) activity. Butanol, ethanol, and water extracts of C. pyxidata DGUM 29005 mycelia were shown to inhibit AChE activity by 99.3%, 93.7%, and 91.7%, respectively. The inhibitory value of the butanol extract was more profound than that of tacrine (95.4%). The ethanol extract also exerted an inhibitory effect against BACE activity; this fraction may harbor the potential for development into a pharmocotherapeutic modality for the treatment of Alzheimer's disease (AD) patients. Rat pheochromocytoma PC12 cells in culture were not determined to be susceptible to the cytotoxic activity evidenced by the mycelial extract. The ethanol extract inhibited endogenous AChE activity in PC12 cellular homogenates, with an IC50 of 67.5 microg/ml, after incubation with intact cells, and also inhibited BACE activity in a dose-dependent fashion. These results suggest that the C. pyxidata mycelial extract has the potential to enhance cholinergic function and, therefore, may perform a function in the amelioration of the cholinergic deficit observed in cases of AD, as well as other types of age associated memory impairment. PMID- 17082744 TI - Analysis of double stranded DNA-dependent activities of Deinococcus radiodurans RecA protein. AB - In this study, the double-stranded DNA-dependent activities of Deinococcus radiodurans RecA protein (Dr RecA) were characterized. The interactions of the Dr RecA protein with double-stranded DNA were determined, especially dsDNA-dependent ATP hydrolysis by the Dr RecA protein and the DNA strand exchange reaction, in which multiple branch points exist on a single RecA protein-DNA complex. A nucleotide cofactor (ATP or dATP ) was required for the Dr RecA protein binding to duplex DNA. In the presence of dATP, the nucleation step in the binding process occurred more rapidly than in the presence of ATP. Salts inhibited the binding of the Dr RecA protein to double-stranded DNA. Double-stranded DNA dependent ATPase activities showed a different sensitivity to anion species. Glutamate had only a minimal effect on the double-stranded DNA-dependent ATPase activities, up to a concentration of 0.7 M. In the competition experiment for Dr RecA protein binding, the Dr RecA protein manifested a higher affinity to double stranded DNA than was observed for single-stranded DNA. PMID- 17082745 TI - Cloning and sequence analysis of a glyceraldehyde-3-phosphate dehydrogenase gene from Ganoderma lucidum. AB - A cDNA library of Ganoderma lucidum has been constructed using a Zap Express cloning vector. A glyceraldehyde-3-phosphate dehydrogenase gene (gpd) was isolated from this library by hybridization of the recombinant phage clones with a gpd-specific gene probe generated by PCR. By comparison of the cDNA and the genomic DNA sequences, it was found that the complete nucleotide sequence encodes a putative polypeptide chain of 338 amino acids interrupted by 6 introns. The predicted amino acid sequence of this gene shows a high degree of sequence similarity to the GPD proteins from yeast and filamentous fungi. The promoter region contains a CT-rich stretch, two CAAT boxes, and a consensus TATA box. The possibility of using the gpd promoter in the construction of new transformation vectors is discussed. PMID- 17082746 TI - Characterization of Osh3, an oxysterol-binding protein, in filamentous growth of Saccharomyces cerevisiae and Candida albicans. AB - OSH3 is one of the seven yeast homologues of the oxysterol binding proteins (OSBPs) which have the major binding affinity to the oxysterols and function as regulator of cholesterol biosynthesis in mammals. Mutational analysis of OSH3 showed that OSH3 plays a regulatory role in the yeast-to-hyphal transition through its oxysterol-binding domain in Saccharomyces cerevisiae. The OSH3 gene was also identified in the pathogenic yeast Candida albicans. Deletion of OSH3 caused a defect in the filamentous growth, which is the major cause of the C. albicans pathogencity. The filamentation defect of the mutation in the MAPK associated transcription factor, namely cph1Delta was suppressed by overexpression of OSH3. These findings suggest the regulatory roles of OSH3 in the yeast filamentous growth and the functional conservations of OSH3 in S. cerevisiae and C. albicans. PMID- 17082747 TI - Isolation and characterization of the smallest bacteriophage P4 derivatives packaged into P4-size head in bacteriophage P2-P4 system. AB - Bacteriophage P4, a satellite phage of coliphage P2, is a very useful experimental tool for the study of viral capsid assembly and cos-cleavage. For an in vitro cos-cleavage reaction study of the P2-P4 system, new shortened and selectable markers containing P4 derivative plasmids were designed as a substrate molecules. They were constructed by swapping the non-essential segment of P4 DNA for either the kanamycin resistance (kmr) gene or the ampicillin resistance (apr) gene. The size of the genomes of the resulting markers were 82% (P4 ash8 delRI:: kmr) and 79% (P4 ash8 delRI:: apr) of the wild type P4 genome. To determine the lower limit of genome size that could be packaged into the small P4-size head, these shortened P4 plasmids were converted to phage particles with infection of the helper phage P2. The conversion of plasmid P4 derivatives to bacteriophage particles was verified by the heat stability test and the burst size determination experiment. CsCl buoyant equilibrium density gradient experiments confirmed not only the genome size of the viable phage form of shortened P4 derivatives, but also their packaging into the small P4-size head. P4 ash8 delRI:: apr turned out to be the smallest P4 genome that can be packaged into P4 sized head. PMID- 17082748 TI - Vibrio vulnificus metalloprotease VvpE has no direct effect on iron-uptake from human hemoglobin. AB - This study was designed to determine whether or not Vibrio vulnificus metalloprotease VvpE can promote iron uptake via the proteolytic cleavage of human hemoglobin. We found that V. vulnificus utilized hemoglobin as an iron source more efficiently via the vulnibactin-mediated iron-uptake system than via the HupA-mediated iron-uptake system and, of the proteases produced by V. vulnificus, VvpE was found to be the only protease capable of destroying hemoglobin. However, VvpE expression, on both the transcriptional and protein levels, was suppressed in iron-limited media. However, vvpE transcription, but not extracellular VvpE production, was reactivated by the addition of hemoglobin or inorganic iron into iron-limited media. Moreover, vvpE transcription began only in the late growth phase when V. vulnificus had already consumed most of the iron for growth. In addition, neither vvpE mutation nor in trans vvpE complementation affected the ability of V. vulnificus to acquire iron or to grow in iron-limited media or in cirrhotic ascites containing hemoglobin. Hemoglobin added into iron-limited media was not destroyed, but gradually formed an insoluble aggregate during culture; this aggregation of hemoglobin occurred regardless of vvpE mutation or complementation. These results indicate that VvpE is not required for efficient iron uptake from hemoglobin. On the contrary, hemoglobin or iron is required for efficient vvpE transcription. In addition, a discrepancy exists between vvpE transcription and extracellular VvpE production in iron-limited media containing inorganic iron or hemoglobin, which suggests that additional unknown posttranscriptional events may be involved in the extracellular production of VvpE. PMID- 17082749 TI - Protective immune response of bacterially-derived recombinant FaeG in piglets. AB - FaeG is the key factor in the infection process of K88ad enterotoxigenic Escherichia coli(ETEC) fimbrial adhesin. In an attempt to determine the possibility of expressing recombinant FaeG with immunogenicity for a new safe and high-production vaccine in E. coli, we constructed the recombinant strain, BL21 (DE3+K88), which harbors an expression vector with a DNA fragment of faeG, without a signal peptide. Results of 15% SDS-polyacrylamide slab gel analysis showed that FaeG can be stably over-expressed in BL21 (DE3+K88) as inclusion bodies without FaeE. Immunoglobulin G (IgG) and M (IgM) responses in pregnant pigs, with boost injections of the purified recombinant FaeG, were detected 4 weeks later in the sera and colostrum. An in vitro villius-adhesion assay verified that the elicited antibodies in the sera of vaccinated pigs were capable of preventing the adhesion of K88ad ETEC to porcine intestinal receptors. The protective effect on the mortality rates of suckling piglets born to vaccinated mothers was also observed one week after oral challenge with the virulent ETEC strain, C83907 (K88ad, CT+, ST+). The results of this study proved that the adhesin of proteinaceous bacterial fimbriae or pili could be overexpressed in engineered E. coli strains, with protective immune responses to the pathogen. PMID- 17082750 TI - Cloning and characterization of a thioredoxin gene, CpTrx1, from the chestnut blight fungus Cryphonectria parasitica. AB - A differential display for the expression profiles of wild-type Cryphonectria parasitica and its virally-infected isogenic hypovirulent strain revealed several transcripts of interest, which evidenced significant matches with fungal genes of known function. Among which, we have further analyzed an amplified PCR product with significant sequence similarity to the known fungal stress-responsive thioredoxin gene from Neurospora crassa. The product of the cloned thioredoxin gene, CpTrx1, consists of 117 amino acids, with a predicted molecular mass of 13.0 kDa and a pI of 5.4. Sequence comparisons demonstrated that the deduced protein sequence of the CpTrx1 gene evidenced a high degree of homology to all known thioredoxins, with the highest degree of homology with trx1, a thioredoxin gene from Saccharomyces cerevisiae, and evidenced a preservation of the conserved hall mark-residues (Trp-Cys-Gly-Pro-Cys) at the active site of thioredoxin. The E. coli-generated CpTRX1 manifested thioredoxin activity, according to the insulin reduction assay, which indicates that the cloned gene does indeed encode for the C. parasitica thioredoxin. PMID- 17082751 TI - Simple method for a cell count of the colonial Cyanobacterium, Microcystis sp. AB - The cell counting of colonial Microcystis spp. is a rather difficult and error prone proposition, as this genus forms irregularly-shaped and irregularly-sized colonies, which are packed with cells. Thus, in order to facilitate a cell count, four methods of dividing the colonies into single cells were compared, including vortexing, sonication, TiO2 treatment, and boiling. As a result, the boiling method was determined to generate the greatest number of single cells from a colony, and all colonies were found to have divided completely after only 6 min of treatment. Furthermore, no significant cell destruction, which might alter the actual cell density, was detected in conjunction with the boiling method (P = 0.158). In order to compute the cell number more simply, the relationship between the colony size and the cell number was determined, via the boiling method. The colony volume, rather than the area or diameter was correlated more closely with the cell number (r2 = 0.727), thereby suggesting that the cell numbers of colonial Microcystis sp. can also be estimated effectively from their volumes. PMID- 17082752 TI - Archaeal communities in mangrove soil characterized by 16S rRNA gene clones. AB - An archaeal 16S rRNA gene library was constructed from mangrove soil. Phylogenetic analysis revealed archaea in mangrove soil including the Crenarchaeota (80.4%) and Euryarchaeota (19.6%) phyla. The archaeal community in mangrove soil appears to be a mixture of organisms found in a variety of environments with the majority being of marine origin. PMID- 17082753 TI - Bacterial diversity in the human saliva from different ages. AB - To obtain primary idea on oral bacterium species that are generally present in periodotally healthy Koreans, the oral bacterial flora in the saliva of four periodontally healthy Koreans at different ages (5, 32, 35, 65) was investigated in this study. For this investigation, 16SrRNA gene clone libraries were generated from the saliva of the four healthy Koreans, and 50 clones were randomly selected from each saliva clone library and sequenced. Totally, 37 different kinds of bacterial 16S rRNA gene sequences were identified based on sequence homology search through GenBank database. The 37 kinds of saliva clone sequences were classified to 14 genera and 2 uncultured and 1 unidentified bacteria. Among the 14 identified genera, Streptococcus, Prevotella, and Veillonella were common genera, and Streptococcus was dominant genus that accounted for 7 different species. Among the seven Streptococcus species, S. salivarius appeared as the most common species. More numbers of species belonging to the genera Streptococcus and Prevotella was present in saliva from ages 32 and 35. While saliva from ages 5 and 65 showed more numbers of species belonging to the genera Rothia, including potential pathogenic species. Overall, saliva of a young child and a senior showed higher bacterial diversity than that of young adults. PMID- 17082754 TI - Nucleocapsid amino acids 211 to 254, in particular, tetrad glutamines, are essential for the interaction between the nucleocapsid and membrane proteins of SARS-associated coronavirus. AB - GST pull-down assays were used to characterize the SARS-CoV membrane (M) and nucleocapsid (N) interaction, and it was found that the amino acids 211-254 of N protein were essential for this interaction. When tetrad glutamines (Q) were replaced with glutamic acids (E) at positions of 240-243 of the N protein, the interaction was disrupted. PMID- 17082756 TI - 25-Hydroxyvitamin D, insulin resistance, and kidney function in the Third National Health and Nutrition Examination Survey. AB - Previous research has reported reduced serum 25-hydroxyvitamin D levels in patients with chronic kidney disease (CKD), although the relationship between vitamin D status and insulin resistance (IR) in patients with CKD has not been examined in the general population. We examined the association that kidney function, based on glomerular filtration rate (eGFR) estimated from serum creatinine, has with serum levels of 25-hydroxyvitamin D and components of the metabolic syndrome among 14 679 participants in the Third National Health and Nutrition Examination Survey (NHANES III). In this analysis, adjusted mean serum 25-hydroxyvitamin D was significantly lower only in the participants with a severe (15-29 ml/min/1.73 m(2)) decrease in eGFR compared to those with normal kidney function (61.6 vs 73.3 nmol/l, P=0.0063). Serum 25-hydroxyvitamnin D (P=0.0018) and level of kidney function (P=0.0003) were inversely associated, independent of each other, with homeostasis model assessment of insulin resistance (HOMA-IR), adjusting for confounders. Participants with high 25 hydroxyvitamin D levels (>81 nmol/l) had lower HOMA-IR. We conclude that 25 hydroxyvitamin D deficiency is not as prevalent in the US general population with decreased eGFR as previously reported in patients with CKD; and that vitamin D and kidney function have independent inverse associations with IR. PMID- 17082757 TI - Plasma creatinine determination in mice and rats: an enzymatic method compares favorably with a high-performance liquid chromatography assay. AB - The use of the colorimetric Jaffe method for the measurement of creatinine in mouse and rat plasma has been criticized as prior studies have shown a dramatic overestimation. We compared a colorimetric picric acid, an enzymatic, and a high performance liquid chromatography (HPLC) method to assess their appropriateness for routine measurements of creatinine in plasma of healthy and diseased mice (n=61) and rats (n=56). For the colorimetric Jaffe method a pronounced overestimation is confirmed. Additionally the method showed interference with hemoglobin already in a very low, non-visible concentration range in rat plasma. The enzymatic measurement demonstrated a hemoglobin interference in mice, only when hemolysis was visible. The comparison between HPLC and the enzymatic measurement gave a good agreement between both methods in both species. Therefore the enzymatic method fulfills the requirements for a routine screening test for plasma creatinine in healthy as well as diseased mice and rats Kiover a broad concentration range. PMID- 17082758 TI - Accepting prospective kidney donors with asymptomatic urinary abnormalities: are we shooting in the dark? PMID- 17082759 TI - Modulation of the human cytokine response by interferon lambda-1 (IFN-lambda1/IL 29). AB - The interferon lambda family (IFN-lambda1/2/3) is a newly described group of cytokines that are related to both the type-1 interferons and IL-10 family members. These novel cytokines are induced during viral infection and, like type 1 interferons, display significant anti-viral activity. In order to understand their function in more depth, we have examined the ability of IFN-lambda1/IL-29 to regulate cytokine production by human immune cells. Whole peripheral blood mononuclear cells (PBMC) exposed to IFN-lambda1 specifically upregulated IL-6, -8 and -10 but there were no visible effects on TNF or IL-1. This response was produced in a dose-dependant fashion and was inhibited by IL-10. Examination of purified cell populations isolated from PBMC demonstrated that monocytes, rather than lymphocytes, were the major IFN-lambda1-responsive cellular subset, producing IL-6, -8 and -10 in response to IFN-lambda1. Monocyte responses induced by low-level LPS stimulation were also synergistically enhanced by the presence of IFN-lambda1. Human macrophages were also shown to react to IFN-lambda1 similarly to monocytes, by producing the cytokines IL-6, -8 and -10. In conclusion, we have shown that IFN-lambda1, a cytokine produced in response to viral infection, activates both monocytes and macrophages producing a restricted panel of cytokines and may therefore be important in activating innate immune responses at the site of viral infection. PMID- 17082760 TI - Association study between the CX3CR1 gene and asthma. AB - CX3CR1, a fractalkine receptor, mediates cell-adhesive and migratory functions in inflammation. Based on CX3CR1 expression observed in bronchial tissues of asthmatic subjects, we hypothesized that genetic variation at this locus may affect susceptibility to asthma. We carried out an association study and a haplotypic analysis with selected polymorphisms of the CX3CR1 in a familial asthmatic sample from a founder population. Genetic analyses performed by FBAT software showed five CX3CR1 single nucleotide polymorphisms (rs938203, rs2669849, rs1050592, T280M and V249I) with significant associations between their common alleles and asthma (P<0.004) in a dominant model. A haplotype formed with common alleles of rs1050592, T280M and V249I is also overtransmitted in asthmatic subjects (P=0.005) under a dominant model. The associations of V249I and rs2669849 have been validated in an independent case-control sample. For V249I, odds ratios (OR) are 2.16 (common homozygous) and 2.11 (heterozygous) in dominant model (P=0.031). For rs2669849, OR are 2.75 (common homozygous) and 1.86 (heterozygous) in additive model (P=0.007) and dominant model (P=0.059). These results suggest an asthma protective effect of the minor alleles in healthy control carriers. Further functional studies of CX3CR1 are needed to document its role in the pathophysiology of asthma. PMID- 17082761 TI - Annexin2 coating the surface of enlargeosomes is needed for their regulated exocytosis. AB - Enlargeosomes are small cytoplasmic vesicles that undergo rapid, Ca2+-dependent exo/endocytosis. The role of the cytoskeleton in these processes was unknown. In PC12-27 cells, microtubule disassembly had little effect on enlargeosomes, whereas microfilament disassembly increased markedly both their resting and stimulated exocytosis, and inhibited their endocytosis. Even at rest enlargeosomes are coated at their cytosolic surface by an actin-associated protein, annexin2, bound by a dual, Ca2+-dependent and Ca2+-independent mechanism. In contrast, the other enlargeosome marker, desmoyokin/Ahnak, is transported across the organelle membrane, apparently by an ABC transporter, and binds to its lumenal face. Annexin2-GFP expression revealed that, upon stimulation, the slow and random enlargeosome movement increases markedly and becomes oriented toward the plasma membrane. After annexin2 downregulation enlargeosome exocytosis induced by both [Ca2+]i rise and cytoskeleton disruption is inhibited, and the NGF-induced differentiation is blocked. Binding of annexin2 to the enlargeosome membrane, the most extensive ever reported (>50% annexin2 bound to approximately 3% of total membrane area), seems therefore to participate in the regulation of their exocytosis. PMID- 17082762 TI - Yeast Pth2 is a UBL domain-binding protein that participates in the ubiquitin proteasome pathway. AB - Ubiquitin-like (UBL)-ubiquitin-associated (UBA) proteins such as Rad23 and Dsk2 mediate the delivery of polyubiquitinated proteins to the proteasome in the ubiquitin-proteasome pathway. We show here that budding yeast peptidyl-tRNA hydrolase 2 (Pth2), which was previously recognized as a peptidyl-tRNA hydrolase, is a UBL domain-binding protein that participates in the ubiquitin-proteasome pathway. Pth2 bound to the UBL domain of both Rad23 and Dsk2. Pth2 also interacted with polyubiquitinated proteins through the UBA domains of Rad23 and Dsk2. Pth2 overexpression caused an accumulation of polyubiquitinated proteins and inhibited the growth of yeast. Ubiquitin-dependent degradation was accelerated in the pth2Delta mutant and was retarded by overexpression of Pth2. Pth2 inhibited the interaction of Rad23 and Dsk2 with the polyubiquitin receptors Rpn1 and Rpn10 on the proteasome. Furthermore, Pth2 function involving UBL-UBA proteins was independent of its peptidyl-tRNA hydrolase activity. These results suggest that Pth2 negatively regulates the UBL-UBA protein-mediated shuttling pathway in the ubiquitin-proteasome system. PMID- 17082763 TI - TRPC3 and TRPC6 are essential for angiotensin II-induced cardiac hypertrophy. AB - Angiotensin (Ang) II participates in the pathogenesis of heart failure through induction of cardiac hypertrophy. Ang II-induced hypertrophic growth of cardiomyocytes is mediated by nuclear factor of activated T cells (NFAT), a Ca(2+)-responsive transcriptional factor. It is believed that phospholipase C (PLC)-mediated production of inositol-1,4,5-trisphosphate (IP(3)) is responsible for Ca(2+) increase that is necessary for NFAT activation. However, we demonstrate that PLC-mediated production of diacylglycerol (DAG) but not IP(3) is essential for Ang II-induced NFAT activation in rat cardiac myocytes. NFAT activation and hypertrophic responses by Ang II stimulation required the enhanced frequency of Ca(2+) oscillation triggered by membrane depolarization through activation of DAG-sensitive TRPC channels, which leads to activation of L-type Ca(2+) channel. Patch clamp recordings from single myocytes revealed that Ang II activated DAG-sensitive TRPC-like currents. Among DAG-activating TRPC channels (TRPC3, TRPC6, and TRPC7), the activities of TRPC3 and TRPC6 channels correlated with Ang II-induced NFAT activation and hypertrophic responses. These data suggest that DAG-induced Ca(2+) signaling pathway through TRPC3 and TRPC6 is essential for Ang II-induced NFAT activation and cardiac hypertrophy. PMID- 17082764 TI - Reversible, cooperative reactions of yeast vacuole docking. AB - Homotypic yeast vacuole fusion occurs in three stages: (i) priming reactions, which are independent of vacuole clustering, (ii) docking, in which vacuoles cluster and accumulate fusion proteins and fusion regulatory lipids at a ring shaped microdomain surrounding the apposed membranes of each docked vacuole, where fusion will occur, and (iii) bilayer fusion/compartment mixing. These stages require vacuolar SNAREs, SNARE-chaperones, GTPases, effector complexes, and chemically minor but functionally important lipids. For each, we have developed specific ligands that block fusion and conditions that reverse each block. Using them, we test whether docking entails a linearly ordered series of catalytic events, marked by sequential acquisition of resistance to inhibitors, or whether docking subreactions are cooperative and/or reversible. We find that each fusion protein and regulatory lipid is needed throughout docking, indicative of a reversible or highly cooperative assembly of the fusion-competent vertex ring. In accord with this cooperativity, vertices enriched in one fusion catalyst are enriched in others. Docked vacuoles finally assemble SNARE complexes, yet still require physiological temperature and lipid rearrangements to complete fusion. PMID- 17082765 TI - Conditional deletion of Nbs1 in murine cells reveals its role in branching repair pathways of DNA double-strand breaks. AB - NBS1 forms a complex with MRE11 and RAD50 (MRN) that is proposed to act on the upstream of two repair pathways of DNA double-strand break (DSB), homologous repair (HR) and non-homologous end joining (NHEJ). However, the function of Nbs1 in these processes has not fully been elucidated in mammals due to the lethal phenotype of cells and mice lacking Nbs1. Here, we have constructed mouse Nbs1 null embryonic fibroblasts and embryonic stem cells, through the Cre-loxP and sequential gene targeting techniques. We show that cells lacking Nbs1 display reduced HR of the single DSB in chromosomally integrated substrate, affecting both homology-directed repair (HDR) and single-stranded annealing pathways, and, surprisingly, increased NHEJ-mediated sequence deletion. Moreover, focus formation at DSBs and chromatin recruitment of the Nbs1 partners Rad50 and Mre11 as well as Rad51 and Brca1 are attenuated in these cells, whereas the NHEJ molecule Ku70 binding to chromatin is not affected. These data provide a novel insight into the function of MRN in the branching of DSB repair pathways. PMID- 17082766 TI - Novel features of the rotary catalytic mechanism revealed in the structure of yeast F1 ATPase. AB - The crystal structure of yeast mitochondrial F(1) ATPase contains three independent copies of the complex, two of which have similar conformations while the third differs in the position of the central stalk relative to the alpha(3)beta(3) sub-assembly. All three copies display very similar asymmetric features to those observed for the bovine enzyme, but the yeast F(1) ATPase structures provide novel information. In particular, the active site that binds ADP in bovine F(1) ATPase has an ATP analog bound and therefore this structure does not represent the ADP-inhibited form. In addition, one of the complexes binds phosphate in the nucleotide-free catalytic site, and comparison with other structures provides a picture of the movement of the phosphate group during initial binding and subsequent catalysis. The shifts in position of the central stalk between two of the three copies of yeast F(1) ATPase and when these structures are compared to those of the bovine enzyme give new insight into the conformational changes that take place during rotational catalysis. PMID- 17082767 TI - Ferroportin-mediated mobilization of ferritin iron precedes ferritin degradation by the proteasome. AB - Ferritin is a cytosolic molecule comprised of subunits that self-assemble into a nanocage capable of containing up to 4500 iron atoms. Iron stored within ferritin can be mobilized for use within cells or exported from cells. Expression of ferroportin (Fpn) results in export of cytosolic iron and ferritin degradation. Fpn-mediated iron loss from ferritin occurs in the cytosol and precedes ferritin degradation by the proteasome. Depletion of ferritin iron induces the monoubiquitination of ferritin subunits. Ubiquitination is not required for iron release but is required for disassembly of ferritin nanocages, which is followed by degradation of ferritin by the proteasome. Specific mammalian machinery is not required to extract iron from ferritin. Iron can be removed from ferritin when ferritin is expressed in Saccharomyces cerevisiae, which does not have endogenous ferritin. Expressed ferritin is monoubiquitinated and degraded by the proteasome. Exposure of ubiquitination defective mammalian cells to the iron chelator desferrioxamine leads to degradation of ferritin in the lysosome, which can be prevented by inhibitors of autophagy. Thus, ferritin degradation can occur through two different mechanisms. PMID- 17082768 TI - Cell entry and cAMP imaging of anthrax edema toxin. AB - The entry and enzymatic activity of the anthrax edema factor (EF) in different cell types was studied by monitoring EF-induced changes in intracellular cAMP with biochemical and microscopic methods. cAMP was imaged in live cells, transfected with a fluorescence resonance energy transfer biosensor based on the protein kinase A regulatory and catalytic subunits fused to CFP and YFP, respectively. The cAMP biosensor was located either in the cytosol or was membrane-bound owing to the addition of a tag determining its myristoylation/palmitoylation. Real-time imaging of cells expressing the cAMP biosensors provided the time course of EF catalytic activity and an indication of its subcellular localization. Bafilomycin A1, an inhibitor of the vacuolar ATPase proton pump, completely prevented EF activity, even when added long after the toxin. The time course of appearance of the adenylate cyclase activity and of bafilomycin A1 action suggests that EF enters the cytosol from late endosomes. EF remains associated to these compartments and its activity shows a perinuclear localization generating intracellular cAMP concentration gradients from the cell centre to the periphery. PMID- 17082769 TI - Rab14 is critical for maintenance of Mycobacterium tuberculosis phagosome maturation arrest. AB - Mycobacterium tuberculosis arrests phagosomal maturation in infected macrophage, and, apart from health significance, provides a superb model system to dissect the phagolysosomal biogenesis pathway. Here, we demonstrate a critical role for the small GTPase Rab14 in maintaining mycobacterial phagosome maturation block. Four-dimensional microscopy showed that phagosomes containing live mycobacteria accumulated Rab14 following phagocytosis. The recruitment of Rab14 had strong functional consequence, as a knockdown of endogenous Rab14 by siRNA or overexpression of Rab14 dominant-negative mutants (Rab14S25N and Rab14N125I) released the maturation block and allowed phagosomes harboring live mycobacteria to progress into phagolysosomes. Conversely, overexpression of the wild-type Rab14 and the constitutively active mutant Rab14Q70L prevented phagosomes with dead mycobacteria from undergoing default maturation into phagolysosomal organelles. Mechanistic studies demonstrated a role for Rab14 in stimulating organellar fusion between phagosomes and early endosomes but not with late endosomes. Rab14 enables mycobacterial phagosomes to maintain early endosomal characteristics and avoid late endosomal/lysosomal degradative components. PMID- 17082770 TI - CAP interacts with cytoskeletal proteins and regulates adhesion-mediated ERK activation and motility. AB - CAP/Ponsin belongs to the SoHo family of adaptor molecules that includes ArgBP2 and Vinexin. These proteins possess an N-terminal sorbin homology (SoHo) domain and three C-terminal SH3 domains that bind to diverse signaling molecules involved in a variety of cellular processes. Here, we show that CAP binds to the cytoskeletal proteins paxillin and vinculin. CAP localizes to cell-extracellular matrix (ECM) adhesion sites, and this process requires binding to vinculin. Overexpression of CAP induces the aggregation of paxillin, vinculin and actin at cell-ECM adhesion sites. Moreover, CAP inhibits adhesion-dependent processes such as cell spreading and focal adhesion turnover, whereas a CAP mutant that is unable to localize to cell-ECM adhesion sites is incapable of exerting these effects. Finally, depletion of CAP by siRNA-mediated knockdown leads to enhanced cell spreading, migration and the activation of the PAK/MEK/ERK pathway in REF52 cells. Taken together, these results indicate that CAP is a cytoskeletal adaptor protein involved in modulating adhesion-mediated signaling events that lead to cell migration. PMID- 17082771 TI - Identification of substrates of the Mycobacterium tuberculosis proteasome. AB - The putative proteasome-associated proteins Mpa (Mycobaterium proteasomal ATPase) and PafA (proteasome accessory factor A) of the human pathogen Mycobacterium tuberculosis (Mtb) are essential for virulence and resistance to nitric oxide. However, a direct link between the proteasome protease and Mpa or PafA has never been demonstrated. Furthermore, protein degradation by bacterial proteasomes in vitro has not been accomplished, possibly due to the failure to find natural degradation substrates or other necessary proteasome co-factors. In this work, we identify the first bacterial proteasome substrates, malonyl Co-A acyl carrier protein transacylase and ketopantoate hydroxymethyltransferase, enzymes that are required for the biosynthesis of fatty acids and polyketides that are essential for the pathogenesis of Mtb. Maintenance of the physiological levels of these enzymes required Mpa and PafA in addition to proteasome protease activity. Mpa levels were also regulated in a proteasome-dependent manner. Finally, we found that a conserved tyrosine of Mpa was essential for function. Thus, these results suggest that Mpa, PafA, and the Mtb proteasome degrade bacterial proteins that are important for virulence in mice. PMID- 17082772 TI - Bacterial outer membrane secretin PulD assembles and inserts into the inner membrane in the absence of its pilotin. AB - Dodecamerization and insertion of the outer membrane secretin PulD is entirely determined by the C-terminal half of the polypeptide (PulD-CS). In the absence of its cognate chaperone PulS, PulD-CS and PulD mislocalize to the inner membrane, from which they are extractable with detergents but not urea. Electron microscopy of PulD-CS purified from the inner membrane revealed apparently normal dodecameric complexes. Electron microscopy of PulD-CS and PulD in inner membrane vesicles revealed inserted secretin complexes. Mislocalization of PulD or PulD-CS to this membrane induces the phage shock response, probably as a result of a decreased membrane electrochemical potential. Production of PulD in the absence of the phage shock response protein PspA and PulS caused a substantial drop in membrane potential and was lethal. Thus, PulD-CS and PulD assemble in the inner membrane if they do not associate with PulS. We propose that PulS prevents premature multimerization of PulD and accompanies it through the periplasm to the outer membrane. PulD is the first bacterial outer membrane protein with demonstrated ability to insert efficiently into the inner membrane. PMID- 17082773 TI - JNK- and Fos-regulated Mmp1 expression cooperates with Ras to induce invasive tumors in Drosophila. AB - Loss of the epithelial polarity gene scribble in clones of Drosophila imaginal disc cells can cooperate with Ras signaling to induce malignant tumors. Such mutant tissue overproliferates, resists apoptosis, leaves its place of origin and invades other organs, ultimately causing lethality. We show that increased Jun N terminal kinase (JNK) signaling resulting from the loss of scribble promotes the movement of transformed cells to secondary sites. This effect requires Fos dependent transcriptional activation of a matrix metalloprotease gene mmp1 downstream of JNK. Expression of the Mmp inhibitor Timp or Mmp RNAi knockdown suppresses cell invasiveness. The proinvasive function of the JNK pathway is revealed in a tumor context when active Ras signaling prevents the apoptotic response to JNK activity as it occurs in nontransformed cells. Based on these results, we present a model that explains the oncogenic cooperation between JNK and Ras, and describes how aberrant regulation of cell survival, proliferation and mobilization cooperate to incite malignant tumor formation. PMID- 17082774 TI - ADAMTS1 mediates the release of antiangiogenic polypeptides from TSP1 and 2. AB - Matrix metalloproteases regulate both physiological and pathological events by processing matrix proteins and growth factors. ADAMTS1 in particular is required for normal ovulation and renal function and has been shown to modulate angiogenesis. Here we report that TSP1 and 2 are substrates of ADAMTS1. Using a combination of mass spectrometry and Edman degradation, we mapped the cleavage sites and characterized the biological relevance of these processing events. ADAMTS1 cleavage mediates the release of polypeptides from the trimeric structure of both TSP1 and 2 generating a pool of antiangiogenic fragments from matrix bound thrombospondin. Using neo-epitope antibodies we confirmed that processing occurs during wound healing of wild-type mice. However, TSP1 proteolysis is decreased or absent in ADAMTS1 null mice; this is associated with delayed wound closure and increased angiogenic response. Finally, TSP1-/- endothelial cells revealed that the antiangiogenic response mediated by ADAMTS1 is greatly dependent on TSP1. These findings have unraveled a mechanistic explanation for the angiostatic functions attributed to ADAMTS1 and demonstrated in vivo processing of TSP1 under situations of tissue repair. PMID- 17082776 TI - PPARbeta-mediated growth suppression of baicalein and dexamethasone in human myeloma cells. PMID- 17082775 TI - Correction of aberrant imprinting of IGF2 in human tumors by nuclear transfer induced epigenetic reprogramming. AB - Loss of genomic imprinting of insulin-like growth factor II (IGF2) is a hallmark of many human neoplasms. We attempted to correct this aberrant epigenotype by transferring nuclei from human tumor cells that showed loss of IGF2 imprinting into enucleated mouse and human fibroblasts that had maintained normal IGF2 imprinting. After nuclear transfer, the abnormal biallelic expression of IGF2 in tumor nuclei transiently converted to normal monoallelic imprinted expression in the reconstructed diploid cells. In tetraploid hybrid cells, however, normal IGF2 imprinting was permanently restored in the tumor genome. Inhibition of the synthesis of putative trans imprinting factors with cycloheximide led to loss of IGF2 imprinting in normal cultured fibroblasts, suggesting that normal cells produce proteins that act in trans to induce or maintain genomic imprinting. These data demonstrate that an abnormal tumor epigenotype can be corrected by in vitro reprogramming, and suggest that loss of imprinting is associated with the loss of activity of non-CTCF trans imprinting factor(s) that are either inactivated or mutated in tumors. PMID- 17082777 TI - Treatment of P190 Bcr/Abl lymphoblastic leukemia cells with inhibitors of the serine/threonine kinase CK2. PMID- 17082778 TI - ZAP-70 expression, as detected by immunohistochemistry on bone marrow biopsies from early-phase CLL patients, is a strong adverse prognostic factor. AB - Zeta-associated protein-70 (ZAP-70), mostly assessed by flow-cytometry (FC), recently emerged as reliable prognostic factor in chronic lymphocytic leukaemia (CLL) at presentation. We evaluated ZAP-70 expression in 156 CLL patients by immunohistochemistry (IHC) on formalin-fixed bone marrow (BM) biopsies at diagnosis. At presentation, 117 patients (75%) were with Binet stage A, 27 (17%) stage B and 12 (8%) stage C. Median follow-up was 61 months (range 6-242). ZAP-70 was expressed in neoplastic lymphocytes of 69 patients (44%). Concordance between ZAP-70 by IHC and ZAP-70 by FC, immunoglobulin heavy chain variable genes (IGHV) mutational status and CD38 expression was found in 41/46 (89%), 41/49 (80%) and in 60/88 (68%) tested cases, respectively. ZAP-70 expression significantly correlated with advanced Binet stage (B-C), diffuse BM infiltration, increased lactate dehydrogenase (LDH) and beta2-microglobulin serum levels and lymphocyte doubling time <12 months. ZAP-70 positivity was significantly related to poorer time to progression (median 16 months vs 158 of ZAP-70-negative cases) (P<0.0001) and overall survival (median 106 months vs not reached) (P=0.0002); this correlation was confirmed at multivariate analysis. ZAP-70 expression correlated with poorer outcome also when evaluated only in the 117 stage A patients. In conclusion, immunohistological detection of ZAP-70 on formalin-fixed BM biopsies at diagnosis appears a useful methodological approach to identify patients with poor prognosis in CLL. PMID- 17082779 TI - Intestinal mucosal dysfunction and infection during remission-induction therapy for acute myeloid leukaemia. AB - Intestinal barrier function was prospectively examined in the course of a clinical trial evaluating the efficacy and safety of lisofylline for reducing cytotoxic therapy-induced intestinal epithelial damage-related infectious morbidity in patients receiving standard remission-induction therapy for acute myeloid leukaemia. The absorption and permeation of oral D-Xylose, lactulose and mannitol were measured weekly from baseline until marrow recovery in adult recipients of idarubicin plus cytarabine for untreated acute myeloid leukaemia. These studies were correlated with non-haematologic chemotherapy-related toxicities reflecting mucosal damage, including nausea, vomiting, stomatitis, diarrhoea, abdominal pain and systemic infection. D-xylose absorption decreased and lactulose:mannitol ratio reflecting intestinal permeability increased from baseline until the second and third week after the beginning of the treatment followed by recovery. These measures correlated with infection rates, nausea, vomiting, diarrhoea and increased blood product utilization. Lisofylline was associated with increased intestinal permeability, nausea, vomiting and infection related morbidity despite a reduction in the duration of neutropaenia. These surrogates of intestinal barrier function correlated well with clinically important outcomes despite the failure to demonstrate reduced morbidity with lisofylline and represent useful objective outcome measurements for future clinical trials of products for the amelioration of the effects of cytotoxic therapy on the intestinal mucosa. PMID- 17082780 TI - Proteomic discovery of Max as a novel interacting partner of C/EBPalpha: a Myc/Max/Mad link. AB - The transcription factor CCAAT/enhancer binding protein a (C/EBPalpha) is important in the regulation of granulopoiesis and is disrupted in human acute myeloid leukemia. In the present study, we sought to identify novel C/EBPalpha interacting proteins in vivo through immunoprecipitation using mass spectrometry based proteomic techniques. We identified Max, a heterodimeric partner of Myc, as one of the interacting proteins of C/EBPalpha in our screen. We confirmed the in vivo interaction of C/EBPalpha with Max and showed that this interaction involves the basic region of C/EBPalpha. Endogenous C/EBPalpha and Max, but not Myc and Max, colocalize in intranuclear structures during granulocytic differentiation of myeloid U937 cells. Max enhanced the transactivation capacity of C/EBPalpha on a minimal promoter. A chromatin immunoprecipitation assay revealed occupancy of the human C/EBPalpha promoter in vivo by Max and Myc under cellular settings and by C/EBPalpha and Max under retinoic acid induced granulocytic differentiation. Interestingly, enforced expression of Max and C/EBPalpha results in granulocytic differentiation of the human hematopoietic CD34(+) cells, as evidenced by CD11b, CD15 and granulocyte colony-stimulating factor receptor expression. Silencing of Max by short hairpin RNA in CD34(+) and U937 cells strongly reduced the differentiation-inducing potential of C/EBPalpha, indicating the importance of C/EBPalpha-Max in myeloid progenitor differentiation. Taken together, our data reveal Max as a novel co-activator of C/EBPalpha functions, thereby suggesting a possible link between C/EBPalpha and Myc-Max-Mad network. PMID- 17082781 TI - Regulation of human epidermal keratinocyte differentiation by the vitamin D receptor and its coactivators DRIP205, SRC2, and SRC3. AB - It has long been known that the active metabolite of vitamin D, 1,25 dihydroxyvitamin D(3), stimulates differentiation and inhibits proliferation in epidermal keratinocytes through interaction with the vitamin D receptor (VDR). VDR functions through the coordinate binding of vitamin D response elements in the DNA and specific coactivator proteins which help to initiate transcription. It was recently observed that VDR binds to two major coactivator complexes, DRIP (VDR-interacting protein) and SRC (steroid receptor coactivator), during keratinocyte differentiation. To determine the role of VDR and its coactivators in mediating keratinocyte differentiation, we developed an adenoviral system to knock down, or in the case of VDR, overexpress these genes. In order to study all stages of keratinocyte development, we employed an advanced differentiated normal human keratinocyte culture system that produces a multilayer phenotype similar to that of normal skin. These studies have shown that VDR, DRIP, and SRC are all required for promotion of both early and late keratinocyte differentiation. Additionally, each individual differentiation marker that was assayed has a different specificity for the coactivators that regulate its expression. PMID- 17082782 TI - DNA-based prenatal diagnosis of harlequin ichthyosis and characterization of ABCA12 mutation consequences. AB - Until the identification of ABCA12 as the causative gene, prenatal diagnosis (PD) for harlequin ichthyosis (HI) had been performed by electron microscopic observation of fetal skin biopsy samples. We report the first case of HI DNA based PD. Direct sequence analysis of ABCA12 revealed that the deceased proband was a compound heterozygote for two novel mutations. The maternal nonsense mutation p.Ser1249Term likely leads to nonsense-mediated messenger RNA decay. The paternal mutation c.7436G>A affects the last codon of exon 50 and was expected to be a splice site mutation. For their third pregnancy, the parents requested PD. Direct sequence analysis of fetal genomic DNA from amniotic fluid cells at 17 weeks gestation revealed the fetus was a compound heterozygote for both mutations. The parents requested the pregnancy to be terminated. Analysis of ABCA12 transcripts of cultured keratinocytes from the abortus showed the presence of six abnormally spliced products from the allele carrying the splice site mutation. Four of them lead to premature termination codons whereas the two others produced shortened proteins missing 21 and 31 amino acids from the second ATP-binding cassette. This report provides evidence for residual ABCA12 expression in HI, and demonstrates the efficiency of early DNA-based PD of HI. PMID- 17082783 TI - Differential regulation of hyaluronan metabolism in the epidermal and dermal compartments of human skin by UVB irradiation. AB - Hyaluronan (HA), a major component of the cutaneous extracellular-matrix, is involved in tissue repair. Human skin is exposed to and damaged by UVB irradiation. Here, we investigate the regulation of HA metabolism in human skin during acute UVB-induced inflammation. Expression of HA synthesizing (HAS) and degrading enzymes hyaluronidase (HYAL) as evaluated by quantitative reverse transcribed PCR in response to UVB differed when fibroblasts and HaCaT keratinocytes, representative cell types in dermis and epidermis, respectively, were compared. Both demonstrated temporally different expression patterns of these genes 3- and 24-hours post-irradiation. This resulted 24-hours post irradiation in an increase in HAS gene expression in both fibroblasts and HaCaT keratinocytes, and an increase in HYAL expression only in fibroblasts. HA production as analyzed by the HA content of conditioned medium was reduced in HaCaT and fibroblast cultures 3-hours post-irradiation, whereas HA increased in HaCaT-cultures 24-hours post-irradiation but remained suppressed in fibroblasts cultures. Consistently, immunohistochemical staining for HA in human skin 24 hours post-irradiation demonstrated an increased epidermal HA, but a decrease in the dermal compartment. Moreover, analysis of the HA content of dermal microdialysis-fluid revealed increased accumulation of HA degradation products 24 hours post-irradiation. These data demonstrate that there is a complex temporal and spatial regulation of HA-metabolism in skin in response to UVB irradiation. PMID- 17082784 TI - The detrimental effects of daily sub-erythemal exposure on human skin in vivo can be prevented by a daily-care broad-spectrum sunscreen. PMID- 17082785 TI - Making the best of an unappealing situation. AB - The appeals process allows authors to ask us to reconsider a negative decision. Here we provide advice on when it's worth appealing and what makes a successful appeal. PMID- 17082786 TI - FtsK: a groovy helicase. PMID- 17082787 TI - RNA ligase does the AMP shuffle. PMID- 17082788 TI - Cotranscriptional splicing regulation: it's not just about speed. PMID- 17082789 TI - DEAD on. PMID- 17082791 TI - Co- and post-translational translocation through the protein-conducting channel: analogous mechanisms at work? AB - Many proteins are translocated across, or integrated into, membranes. Both functions are fulfilled by the 'translocon/translocase', which contains a membrane-embedded protein-conducting channel (PCC) and associated soluble factors that drive translocation and insertion reactions using nucleotide triphosphates as fuel. This perspective focuses on reinterpreting existing experimental data in light of a recently proposed PCC model comprising a front-to-front dimer of SecY or Sec61 heterotrimeric complexes. In this new framework, we propose (i) a revised model for SRP-SR-mediated docking of the ribosome-nascent polypeptide to the PCC; (ii) that the dynamic interplay between protein substrate, soluble factors and PCC controls the opening and closing of a transmembrane channel across, and/or a lateral gate into, the membrane; and (iii) that co- and post translational translocation, involving the ribosome and SecA, respectively, not only converge at the PCC but also use analogous mechanisms for coordinating protein translocation. PMID- 17082793 TI - Human adenovirus type 35 vector for gene therapy of brain cancer: improved transduction and bypass of pre-existing anti-vector immunity in cancer patients. AB - Clinical trials in malignant glioma have demonstrated excellent safety of recombinant adenovirus type 5 (Ad5) but lack of convincing efficacy. The overall low expression levels of the Coxsackie and Adenovirus receptor and the presence of high anti-Ad5-neutralizing antibody (NAb) titers in the human population are considered detrimental for consistency of clinical results. To identify an adenoviral vector better suited to infect primary glioma cells, we tested a library of fiber-chimeric Ad5-based adenoviral vectors on 12 fresh human glioma cell suspensions. Significantly improved marker gene expression was obtained with several Ad5-chimeric vectors, predominantly vectors carrying fiber molecules derived from B-group viruses (Ad11, Ad16, Ad35 and Ad50). We next tested Ad35 sero prevalence in sera derived from 90 Dutch cancer patients including 30 glioma patients and investigated the transduction efficiency of this vector in glioma cell suspensions. Our results demonstrate that the sero prevalence and the titers of NAb against Ad35 are significantly lower than against Ad5. Also, recombinant Ad35 has significantly increased ability to transfer a gene to primary glioma cells compared to Ad5. We thus conclude that Ad35 represents an interesting candidate vector for gene therapy of malignant glioma. PMID- 17082794 TI - Long-term follow-up of gene-marked CD34+ cells after autologous stem cell transplantation for multiple myeloma. AB - Gene marking can be used to investigate if progenitor cells harvested from patients are contaminated with tumorigenic cells. It can also provide information about the contribution of hematopoietic stem cells to long-term engraftment and about long-term transgene expression from integrated retroviral vectors. In order to study autologous-infused cell contribution to relapse as well as the long-term persistence of the transgene in hematopoietic cells following autologous bone marrow (BM) transplantation for multiple myeloma, we genetically marked autologous CD34+ enriched BM or peripheral blood cell grafts of eight myeloma patients using retroviral vectors. Six patients were subsequently transplanted with the marked graft and followed with regular time points of analysis. Briefly, mononuclear cells were harvested by leukapheresis during 2-4 consecutive days following priming with granulocyte-macrophage colony-stimulating factor (GM-CSF) or G-CSF. The CD34+ cells separated on Cellpro ceprate avidin-biotin columns were exposed to the G1Na vector coding for neomycin resistance gene at a ratio of five vector particles per cell at three consecutive time points achieving an average transduction efficacy of 2% (0.43-5.1%). The patients were transplanted with a mixture of transduced cells and un-manipulated graft. Vector integration and transgene expression were analyzed by colony assays and polymerase chain reaction. The transgene could be detected for up to 5 years post-transplant in normal BM cells, even in remission following relapse and no side effects related to retroviral gene transfer were observed. There were no marked myeloma cells observed in the patients either in remission or in relapsing disease, which indicates that contribution of infused cells to relapse is unlikely. PMID- 17082795 TI - A gene transfer comparative study of HSA-conjugated antiangiogenic factors in a transgenic mouse model of metastatic ocular cancer. AB - Different antiangiogenic and antimetastatic recombinant adenoviruses were tested in a transgenic mouse model of metastatic ocular cancer (TRP1/SV40 Tag transgenic mice), which is a highly aggressive tumor, developed from the pigmented epithelium of the retina. These vectors, encoding amino-terminal fragments of urokinase plasminogen activator (ATF), angiostatin Kringles (K1-3), endostatin (ES) and canstatin (Can) coupled to human serum albumin (HSA) were injected to assess their metastatic and antiangiogenic activities in our model. Compared to AdCO1 control group, AdATF-HSA did not significantly reduce metastatic growth. In contrast, mice treated with AdK1-3-HSA, AdES-HSA and AdCan-HSA displayed significantly smaller metastases (1.19+/-1.19, 0.87+/-1.5, 0.43+/-0.56 vs controls 4.04+/-5.12 mm3). Moreover, a stronger inhibition of metastatic growth was obtained with AdCan-HSA than with AdK1-3-HSA (P=0.04). Median survival was improved by 4 weeks. A close correlation was observed between the effects of these viruses on metastatic growth and their capacity to inhibit tumor angiogenesis. Our study indicates that systemic antiangiogenic factors production by recombinant adenoviruses, particularly Can, might represent an effective way of delaying metastatic growth via inhibition of angiogenesis. PMID- 17082796 TI - Persistent mismatch repair deficiency following targeted correction of hMLH1. AB - The use of gene therapy to correct mutated or lost gene function for the treatment of human cancers has been an active, yet problematic area of biomedical research. Many technical difficulties, including efficient tissue-specific delivery, integration site specificity and general toxicity, are being addressed. Little is known, however, about the genetic and phenotypic stability that accompanies a successful gene-specific targeting event in a cancer cell. This question was addressed following the creation of a colon cancer cell line in which a mutated hMLH1 gene was corrected via targeted homologous recombination. This correction resulted in the expression of wild-type hMLH1 protein, restoration of the hPMS2 protein and mismatch repair (MMR) proficiency. One of two hMLH1-corrected clones, however, was found to retain defects in MMR activity. These cells continued to express the corrected hMLH1 protein, but had lost expression of another MMR protein, hMSH6. DNA sequence analysis of the hMSH6 gene revealed biallelic expansions of a cytosine repeat region in exon 5 that result in frameshifts leading to premature stop codons. These findings suggest that, similar to acquired drug resistance, the presence of genetically heterogeneous cancer cell populations or acquisition of compensatory mutations can result in 'resistance' to gene replacement therapy. PMID- 17082797 TI - Arterial elasticity identified by pulse wave analysis and its relation to endothelial function in patients with coronary artery disease. AB - Patients with coronary artery disease (CAD) have impaired endothelial function. Arterial elasticity is modulated by endothelial function. The association between arterial elasticity and endothelial function has not been reported in patients with CAD. The present study was designed to investigate whether endothelial dysfunction contributes to impaired arterial elasticity. Thirty patients with CAD and 30 control subjects were recruited. Large and small artery elasticity indices were non-invasively assessed using pulse wave analysis. Brachial artery endothelium-dependent and -independent function were assessed by vascular response to flow-mediated vasodilation (FMD) and sublingual nitroglyceride (NTG), respectively. C1 large artery elasticity index was not different in the CAD group compared with the control group. However, C2 small artery elasticity index was significantly reduced in the CAD group compared with the control group. Flow mediated vasodilation (FMD) was also impaired in the CAD group compared with the control group. Flow-mediated vasodilation (FMD) in the brachial artery correlated with C2 small arterial elasticity index. But NTG-mediated brachial artery vasodilation was similar between the two groups. The present findings suggest that the patients with CAD have reduced C2 small arterial elasticity index and impaired FMD. Endothelial dysfunction is involved in diminished arterial elasticity, suggesting that C2 small arterial elasticity index is a novel surrogate measure for the clinical evaluation of endothelial function. PMID- 17082798 TI - Influence of age and sex on prevalence of masked hypertension determined from home blood pressure measurements. PMID- 17082799 TI - Is it possible to predict the onset of Graves' disease? PMID- 17082800 TI - Genetic predictors of ovarian hyperstimulation syndrome in women undergoing in vitro fertilization. PMID- 17082801 TI - Management of obesity in Prader-Willi syndrome. PMID- 17082802 TI - Does denosumab improve low BMD in postmenopausal women? PMID- 17082803 TI - HIV and the thyroid--what every practicing endocrinologist needs to know. PMID- 17082804 TI - Are elevated aldosterone levels associated with refractory hypertension? PMID- 17082805 TI - Preventing fractures--how good are we really? PMID- 17082806 TI - What is the pathogenesis of hyponatremia after subarachnoid hemorrhage? PMID- 17082807 TI - How beneficial is radioiodine therapy for treating distant metastases from thyroid carcinoma? PMID- 17082808 TI - The role of oxyntomodulin and peptide tyrosine-tyrosine (PYY) in appetite control. AB - Oxyntomodulin and peptide tyrosine-tyrosine (PYY) are released from intestinal enteroendocrine cells in response to a meal. These circulating hormones are considered to be satiety signals, as they have been found to decrease food intake, body weight and adiposity in rodents. Their effect on energy homeostasis is mediated by the hypothalamus and brainstem, and several studies have demonstrated alterations in neuropeptide signaling within the arcuate nucleus. The weight loss that has been observed in animal models after repeated administration of oxyntomodulin and PYY has led to interest in developing these peptides as antiobesity therapies in humans. Indeed, preliminary studies have found that oxyntomodulin or PYY administration reduces food intake and body weight effectively in overweight human volunteers. This research suggests that modulation of these gut hormones could prove to be effective long-term therapies in the quest to combat the obesity epidemic. PMID- 17082809 TI - Mechanisms of disease: regulation of glucocorticoid and receptor levels--impact on the metabolic syndrome. AB - Glucocorticoids exert their effects in target tissues predominantly through their interaction with the glucocorticoid receptor, a member of the nuclear receptor superfamily of transcription factors. Over the years many studies have linked hormone responsiveness, both in vitro and in vivo, to the levels of both glucocorticoid and glucocorticoid receptor; furthermore, an impact of glucocorticoid receptor subcellular trafficking on hormone response has been revealed. This review will focus on the molecular mechanisms responsible for the regulation of glucocorticoid receptor trafficking and expression, and will highlight work that revealed selective physiological effects of altered glucocorticoid receptor expression. The role of alterations in glucocorticoid levels and glucocorticoid receptor function in the metabolic syndrome will also be discussed. PMID- 17082810 TI - Mechanisms of disease: adrenocortical tumors--molecular advances and clinical perspectives. AB - Most adrenocortical tumors are benign, unilateral, adrenocortical adenomas that are often discovered incidentally. Adrenocortical cancer is rare. Exceptionally, adrenocortical tumors can be bilateral. Although most adrenocortical tumors occur sporadically, they may also feature in congenital and/or familial disease. The identification of germline genetic defects in familial diseases associated with adrenocortical tumors helped to define the somatic alterations in sporadic disease: for example, overexpression of insulin-like growth factor 2 and alterations at the 11p15 locus (observed in Beckwith-Wiedemann syndrome) are also found in most adrenocortical cancers. Similarly, inactivating mutations of the TP53 gene, located at 17p13 (observed in Li-Fraumeni syndrome), can also be found at the somatic level in sporadic adrenocortical cancers, as can 17p13 allelic losses. Components of the cyclic AMP signaling pathway--for example, adrenocorticotropic hormone receptors and other membrane receptors, Gs proteins and protein kinase A--can be altered to various degrees in adrenocortical tumors. More recently, gene profiling and genetic studies have shown that the Wnt-beta catenin signaling pathway is frequently activated in adrenocortical tumors. These research findings already have profound implications for clinical management of patients with adrenocortical tumors, for example in unraveling the genetic origin of the disease in some patients, and in the development of molecular markers for diagnosis and prognosis. The new findings should also help in the development of new therapeutic options. PMID- 17082811 TI - Cushing's syndrome caused by an occult source: difficulties in diagnosis and management. AB - BACKGROUND: A 24-year-old woman presented with a 12.5 kg weight gain over 6 months (mostly abdominal), hirsutism, acne, ankle edema, polydipsia, nocturia, back pain, pigmentation, poor libido and lightened menses to our hospital in May 1986. She had been treated for the previous 2 years with furosemide and spironolactone for peripheral edema, and had stopped the combined oral contraceptive 2 months previously. She did not take tobacco, recreational drugs or alcohol. Upon physical examination she was grossly Cushingoid with florid clinical manifestations. INVESTIGATIONS: Serum potassium and bicarbonate, circadian rhythm of cortisol, low-dose and high-dose dexamethasone suppression tests, plasma adrenocorticotropic hormone (ACTH), corticotropin releasing-hormone stimulation test, CT scan of the pituitary, plain chest radiology, CT scan of the chest and abdomen, trans-sphenoidal pituitary biopsy and histology, CT scan and MRI of the thorax, MRI of the pituitary, octreotide scintigraphy, gastroscopy, colonoscopy, gut peptides, tumor markers, urine 5-hydroxyl-indole-acetic acid, resection, histology, immunocytochemistry and in situ hybridization. DIAGNOSIS: Occult ectopic ACTH syndrome from a presumed appendiceal neuroendocrine tumor. The tumor was only identified some 20 years from initial presentation. MANAGEMENT: Adrenolytic therapy before bilateral adrenalectomy to cure Cushing's syndrome, glucocorticoid and mineralocorticoid replacement therapy, and then repeated surveillance over 20 years to locate the ectopic source of ACTH. This was finally identified by CT scan and excised at laparotomy. PMID- 17082813 TI - Caspase function in programmed cell death. AB - The first proapoptotic caspase, CED-3, was cloned from Caenorhabditis elegans in 1993 and shown to be essential for the developmental death of all somatic cells. Following the discovery of CED-3, caspases have been cloned from several vertebrate and invertebrate species. As reviewed in other articles in this issue of Cell Death and Differentiation, many caspases function in nonapoptotic pathways. However, as is clear from the worm studies, the evolutionarily conserved role of caspases is to execute programmed cell death. In this article, I will specifically focus on caspases that function primarily in cell death execution. In particular, the physiological function of caspases in apoptosis is discussed using examples from the worm, fly and mammals. PMID- 17082814 TI - Caspase substrates. AB - The relatively common occurrence of sequences within proteins that match the consensus substrate specificity of caspases in intracellular proteins suggests a multitude of substrates in vivo - somewhere in the order of several hundred in humans alone. Indeed, the list of proteins that are reported to be cleaved by caspases in vitro proliferates rapidly. However, only a few of these proteins have been rigorously established as biologically or pathologically relevant, bona fide substrates in vivo. Many of them probably simply represent 'innocent bystanders' or erroneous assignments. In this review we discuss concepts of caspase substrate recognition and specificity, give resources for the discovery and annotation of caspase substrates, and highlight some specific human or mouse proteins where there is strong evidence for biologic or pathologic relevance. PMID- 17082815 TI - Requirement for Daxx in mature T-cell proliferation and activation. AB - The protein Daxx promotes Fas-mediated cell death through activation of apoptosis signal-regulating kinase 1, leading to the activation of the MAPKs JNK and p38. Owing to the in utero lethality of daxx-deficient mice, the in vivo role of Daxx has been so far difficult to analyze. We have generated transgenic mice expressing a dominant-negative form of Daxx (Daxx-DN) in the T-cell lineage. We show that Daxx is recruited to the Fas receptor upon FasL engagement and that Daxx-DN expression protects activated T cells from Fas-induced cell death, by preventing the death-inducing signal complex to be properly formed. Normal lymphocyte development and homeostasis are nevertheless observed. Interestingly, we report that both in vitro and in vivo stimulation of Daxx-DN T-lymphocytes leads to increased proliferative T-cell responses. This increased proliferation is associated with a marked increase in tyrosine phosphorylation of LAT and ZAP70 as Daxx-DN favor their recruitment to the T-cell receptor (TCR) complex. These findings identify Daxx as a critical regulator of T-lymphocyte homeostasis by decreasing TCR-induced cell proliferation and by promoting Fas-mediated cell death. PMID- 17082816 TI - CD4/CXCR4-mediated cell death in AIDS. PMID- 17082817 TI - Caspase-2 involvement during ionizing radiation-induced oocyte death in the mouse ovary. AB - In mammals, the pool of primordial follicles at birth is determinant for female fertility. Exposure to IR during oogonia proliferation and the diplotene stages of ovarian development induced the virtual disappearance of primordial follicles in the postnatal ovary, while half the follicular reserve remained present after irradiation during the zygotene/pachytene stages. This sensitivity difference was correlated with the level of caspase-2 expression evaluated by immunohistochemistry. At the diplotene stage, Western blot and caspase activity analysis revealed that caspase-2 was activated 2 h after irradiation and a significant increase in the number of oocytes expressing cleaved caspase-9 and -3 occurred 6 h after treatment. Inhibition of caspase-2 activity prevented the cleavage of caspase-9 and partially prevented the loss of oocytes in response to irradiation. Taken together, our results show that caspase-2-dependent activation of the mitochondrial apoptotic pathway is one of the mechanisms involved in the genotoxic stress-induced depletion of the primordial follicle pool. PMID- 17082818 TI - Epigenetic regulation of transcription in intermediate heterochromatin. AB - Constitutive heterochromatin is a compact, transcriptionally inert structure formed in gene-poor and repeat- and transposon-rich regions. In Arabidopsis, constitutive heterochromatin is characterized by hypermethylated DNA and histone H3 dimethylated at lysine (K) 9 (H3K9me2) together with depletion of histone H3 dimethylated at lysine 4 (H3K4me2). Here, we describe loci with intermediate properties of heterochromatin in which transcription downregulation is inherited in a manner similar to constitutive heterochromatin, although the loci are associated with opposing histone marks--H3K4me2 and H3K9me2. In the ddm1 (decrease in DNA methylation 1) mutants, their transcriptional activation is accompanied by the expected shift in the H3 modifications--depletion of H3K9me2 and enrichment in H3K4me2. In mom1 (Morpheus' molecule 1) mutants, however, a marked increase in transcription is not accompanied by detectable changes in the levels of H3K4me2 and H3K9me2. Therefore, transcriptional regulation in the intermediate heterochromatin involves two distinct epigenetic mechanisms. Interestingly, silent transgenic inserts seem to acquire properties characteristic of the intermediate heterochromatin. PMID- 17082819 TI - Molecular mechanism of P pilus termination in uropathogenic Escherichia coli. AB - P pili are important adhesive fibres that are assembled by the conserved chaperone-usher pathway. During pilus assembly, the subunits are incorporated into the growing fibre by the donor-strand exchange mechanism, whereby the beta strand of the chaperone, which complements the incomplete immunoglobulin fold of each subunit, is displaced by the amino-terminal extension of an incoming subunit in a zip-in-zip-out exchange process that is initiated at the P5 pocket, an exposed hydrophobic pocket in the groove of the subunit. In vivo, termination of P pilus growth requires a specialized subunit, PapH. Here, we show that PapH is incorporated at the base of the growing pilus, where it is unable to undergo donor-strand exchange. This inability is not due to a stronger PapD-PapH interaction, but to a lack of a P5 initiator pocket in the PapH structure, suggesting that PapH terminates pilus growth because it is lacking the initiation point by which donor-strand exchange proceeds. PMID- 17082820 TI - The UBL domain of PLIC-1 regulates aggresome formation. AB - Defects in protein folding and the proteasomal pathway have been linked with many neurodegenerative diseases. PLIC-1 (protein linking IAP to the cytoskeleton) is a ubiquitin-like protein that binds to the ubiquitin-interacting motif (UIM) of the proteasomal subunit S5a. Here, we show that PLIC-1 also binds to the UIM proteins ataxin 3--a deubiquitinating enzyme--HSJ1a--a co-chaperone--and EPS15 (epidermal growth factor substrate 15)--an endocytic protein. Using a polyglutamine (polyQ) disease model, we found that both endogenous PLIC-1 and EPS15 localize to perinuclear aggresomes, and that polyQ enhances their in vivo interaction. We show that knockdown of PLIC-1 and EPS15 by RNA interference reduces aggresome formation. In addition, PLIC-1(DeltaUBL) functions as a dominant-negative mutant, blocking both polyQ transport to aggresomes and the association of EPS15 with dispersed aggregates. We also show that PLIC-1 is upregulated by arsenite-induced protein misfolding. These results indicate a role for PLIC-1 in the protein aggregation-stress pathway, and we propose a novel function for the ubiquitin like (UBL) domain--by means of UBL-UIM interactions--in transport to aggresomes. PMID- 17082821 TI - MOM1 mediates DNA-methylation-independent silencing of repetitive sequences in Arabidopsis. AB - The heterochromatic regions around centromeres of animal and plant chromosomes are composed of tandem repetitive sequences, interspersed with transposons and transposon derivatives. These sequences are largely transcriptionally silent and highly methylated, and are associated with specifically modified histones. Although embedded in heterochromatin, Arabidopsis 5S ribosomal RNA genes are among the most highly transcribed genes. However, some 5S genes are silenced, and we show here that this silencing can be suppressed by a reduction in CG methylation. Importantly, we show that mutation of MORPHEUS' MOLECULE 1 (MOM1) releases 5S repeat silencing independently of chromatin properties, as illustrated by the absence of detectable alteration of DNA and histone H3 methylation patterns. MOM1 also prevents transcription of 180-bp satellite repeats and 106B dispersed repeats but not of transposons. Our results provide evidence that transcription of densely methylated and highly repetitive heterochromatic sequences is controlled by two distinct epigenetic silencing pathways, one dependent on and the other independent of DNA methylation. PMID- 17082822 TI - Cryptic Species in the Anopheles (Nyssorhynchus) albitarsis (Diptera: Culicidae) Complex: Incongruence Between Random Amplified Polymorphic DNA-Polymerase Chain Reaction Identification and Analysis of Mitochondrial DNA COI Gene Sequences. AB - Random amplified polymorphic DNA (RAPD) diagnostic bands are one tool used to differentiate cryptic mosquito species in the Anopheles albitarsis Complex. Monophyly of four species (A. albitarsis Lynch-Arribalzaga, A. albitarsis B, A. deaneorum Rosa-Freitas, and A. marajoara Galvao & Damasceno) currently identified with the RAPD technique was assessed using sequences of the cytochrome oxidase I (COI) mitochondrial DNA (mtDNA) gene. Maximum parsimony, maximum likelihood, and Bayesian analyses support monophyly for A. albitarsis s.s., A. albitarsis B, and A. deaneorum. Anopheles marajoara, as identified by RAPD banding patterns, was either polyphyletic or paraphyletic in all phylogenetic analyses. The phylogenetic pattern and within-species genetic distances observed in A. marajoara suggest the existence of a previously unidentified species (species E) in northern Brazil and Venezuela. Diagnostic RAPD bands were unable to distinguish between A. marajoara and species E, probably because of the low number of correlated bands used to identify species and weaknesses of the RAPD technique, in particular, violations of the untested assumption of homology of comigrating bands. A. marajoara (even without species E) is paraphyletic with respect to A. deaneorum; if A. deaneorum is a separate species from A. marajoara, then A. marajoara may consist of two or more species in Amazonian Brazil. Based on mtDNA COI sequences, there are at least four phylogenetic species within the Albitarsis Complex: A. albitarsis s.s., A. albitarsis B, A. marajoara, and species E; the species status of A. deaneorum is ambiguous. PMID- 17082823 TI - Optical-CT scanning of polymer gels. AB - The application of optical-CT scanning to achieve accurate high-resolution 3D dosimetry is a subject of current interest. The purpose of this paper is to provide a brief overview of past research and achievements in optical-CT polymer gel dosimetry, and to review current issues and challenges. The origins of optical-CT imaging of light-scattering polymer gels are reviewed. Techniques to characterize and optimize optical-CT performance are presented. Particular attention is given to studies of artifacts in optical-CT imaging, an important area that has not been well studied to date. The technique of optical-CT simulation by Monte-Carlo modeling is introduced as a tool to explore such artifacts. New simulation studies are presented and compared with experimental data. PMID- 17082826 TI - Beyond the mere dichotomy of active search versus avoidance of information about the self. PMID- 17082827 TI - Expert synthesis of the literature to support critical care decision making. PMID- 17082828 TI - Randomized trial for answers to clinical questions: evaluating a pre-appraised versus a MEDLINE search protocol. AB - OBJECTIVE: The paper compares the speed, validity, and applicability of two different protocols for searching the primary medical literature. DESIGN: A randomized trial involving medicine residents was performed. SETTING: An inpatient general medicine rotation was used. PARTICIPANTS: Thirty-two internal medicine residents were block randomized into four groups of eight. MAIN OUTCOME MEASURES: Success rate of each search protocol was measured by perceived search time, number of questions answered, and proportion of articles that were applicable and valid. RESULTS: Residents randomized to the MEDLINE-first (protocol A) group searched 120 questions, and residents randomized to the MEDLINE-last (protocol B) searched 133 questions. In protocol A, 104 answers (86.7%) and, in protocol B, 117 answers (88%) were found to clinical questions. In protocol A, residents reported that 26 (25.2%) of the answers were obtained quickly or rated as "fast" (<5 minutes) as opposed to 55 (51.9%) in protocol B, (P = 0.0004). A subset of questions and articles (n = 79) were reviewed by faculty who found that both protocols identified similar numbers of answer articles that addressed the questions and were felt to be valid using critical appraisal criteria. CONCLUSION: For resident-generated clinical questions, both protocols produced a similarly high percentage of applicable and valid articles. The MEDLINE-last search protocol was perceived to be faster. However, in the MEDLINE-last protocol, a significant portion of questions (23%) still required searching MEDLINE to find an answer. PMID- 17082829 TI - Retention of retrospective print journals in the digital age: trends and analysis. AB - PURPOSE: The issue of retaining retrospective print journals is examined in light of the shift to electronic titles, the reallocation of library budgets from print to electronic, and the changing research practices of today's library users. This article also examines the evolving role of the physical library and its impact on space allocation. METHODS: To determine current practice and opinion, a survey of health sciences librarians and academic librarians was conducted. To demonstrate the use patterns of older journal issues, citation analyses and interlibrary loan statistics were examined. RESULTS: All methods indicate that recent material is accessed more frequently than older material, with a significant drop in use of materials greater than 15 years old. Materials greater than 20 years old constituted less than 5% of interlibrary loans and less than 9% of articles noted in the citation analysis. CONCLUSIONS: It is possible to eliminate older years of a print journal collection without a large impact on the needs of researchers. Librarians' preference to maintain full runs of journal titles may be motivated by reasons outside of actual usage or patrons needs. PMID- 17082830 TI - Development of a subject search filter to find information relevant to palliative care in the general medical literature. AB - PURPOSE: The research developed and validated palliative care search filters for use in the general biomedical literature. METHODS: Four general medical journals were hand-searched to identify articles relevant to palliative care, forming a "gold standard" reference set. Searches comprising Medical Subject Headings (MeSH) and text-words were created for use in Ovid MEDLINE, and retrieved references were compared to the gold standard. Sensitivity, specificity, accuracy, and precision rates were calculated. RESULTS: By hand-searching 20,501 articles published in the 4 journals during the 3-year study period (1999-2001), reviewers identified 773 items relevant to palliative care (3.8%). A master search combining 9 MeSH descriptors with 3 text-words achieved 45.4% sensitivity, 99.3% specificity, 73% precision, and 97.3% accuracy. Efforts to increase the sensitivity by modifying 3 relevant published but unvalidated searches did not improve the yield, except in 1 case which resulted in an improved sensitivity of 56.9% but was offset by reduced specificity (92.1%), precision (22%), and accuracy (90.8%). CONCLUSIONS: The study confirmed that literature relevant to palliative care is difficult to identify in general medical journals. While the filter developed in this research represents the best trade-off between sensitivity, specificity, accuracy, and precision, the sensitivity is unacceptably low. Further research, such as frequency analysis of text-words and MeSH terms, is required to increase the sensitivity of searching in this subset of the literature. PMID- 17082831 TI - Evaluation of a liaison librarian program: client and liaison perspectives. AB - OBJECTIVES: This paper describes a survey-based evaluation of the five-year old Liaison Librarian Program at the University of Florida. METHODS: Liaison librarians, faculty, students, staff, residents, and post-doctoral associates were queried via Web-based surveys. Questions addressed client and liaison perspectives on a variety of issues, including program and service awareness and usage, client-library relations and communication, client support for the program, and liaison workload. RESULTS: Approximately 43% of the 323 client respondents were aware of liaison services; 72% (n = 163) of these clients had had contact with their liaison. Ninety-five percent (n = 101) of faculty and students who reported contact with their liaison supported the continuation of the program. Liaison services were used by a greater percentage of faculty than students, although they had similar patterns of usage and reported the same "traditional" services to be most important. Liaisons indicated that communications with clients had increased, the reputation of the library was enhanced, and their workloads had increased as a result of the Liaison Librarian Program. CONCLUSIONS AND RECOMMENDATIONS: Survey results suggest that the Liaison Librarian Program has a core set of clients who use and highly value the services provided by liaisons. Recommendations addressing workload, training, marketing, and administrative support are provided. PMID- 17082832 TI - Mapping of medical acronyms and initialisms to Medical Subject Headings (MeSH) across selected systems. AB - INTRODUCTION: Given the common use of acronyms and initialisms in the health sciences, searchers may be entering these abbreviated terms rather than full phrases when searching online systems. The purpose of this study is to evaluate how various MEDLINE Medical Subject Headings (MeSH) interfaces map acronyms and initialisms to the MeSH vocabulary. METHODS: The interfaces used in this study were: the PubMed MeSH database, the PubMed Automatic Term Mapping feature, the NLM Gateway Term Finder, and Ovid MEDLINE. Acronyms and initialisms were randomly selected from 2 print sources. The test data set included 415 randomly selected acronyms and initialisms whose related meanings were found to be MeSH terms. Each acronym and initialism was entered into each MEDLINE MeSH interface to determine if it mapped to the corresponding MeSH term. Separately, 46 commonly used acronyms and initialisms were tested. RESULTS: While performance differed widely, the success rates were low across all interfaces for the randomly selected terms. The common acronyms and initialisms tested at higher success rates across the interfaces, but the differences between the interfaces remained. CONCLUSION: Online interfaces do not always map medical acronyms and initialisms to their corresponding MeSH phrases. This may lead to inaccurate results and missed information if acronyms and initialisms are used in search strategies. PMID- 17082833 TI - Reporting of non-communicable disease research in low- and middle-income countries: a pilot bibliometric analysis. AB - OBJECTIVE: The paper identifies the relative amount of research devoted to non communicable disease in low- and middle-income countries (LMICs). DESIGN: A bibliometric analysis of a subset of journals published in LMICs was performed. MEASUREMENTS: Seventy-six peer-reviewed journals focused on general medicine or public health published in 46 LMICs and indexed from 1998 to 2003 in MEDLINE. A total of 24 journals were selected, 4 journals from each of 6 LMIC regions. Searches were refined using 18 non-communicable disease topics with 7,012 articles identified for analysis. RESULTS: More than 40% of articles in LMIC regions focused on non-communicable disease research. The percentage was highest in Eastern Europe/Central Asia (47%) and lowest in Latin America (36%). The percentage of articles published in Sub-Saharan Africa (38%) did not differ significantly from that of Latin America or South Asia. Cardiovascular disease and cancer led the list of the top ten most-indexed published topics by region. CONCLUSIONS: Even in regions rampant with infectious diseases, some capability exists to conduct research on non-communicable diseases. Greater attention should be paid to the conduct and support of such research in LMICs, which will benefit these countries and may yield clues to lower-cost solutions to the burden of these diseases worldwide. PMID- 17082835 TI - Bibliometric analysis of the American Journal of Veterinary Research to produce a list of core veterinary medicine journals. AB - OBJECTIVE: Bibliometric techniques were used to analyze the citation patterns of researchers publishing in the American Journal of Veterinary Research (AJVR). METHODS: The more than 25,000 bibliographic references appearing in the AJVR from 2001 to 2003 were examined for material type, date of publication, and frequency of journals cited. Journal titles were ranked in decreasing order of productivity to create a core list of journals most frequently used by veterinary medical researchers. RESULTS: The majority of items cited were journals (88.8%), followed by books (9.8%) and gray literature (2.1%). Current sources of information were favored; 65% of the journals and 77% of the books were published in 1990 or later. Dividing the cited articles into 3 even zones revealed that 24 journals produced 7,361 cited articles in the first zone. One hundred thirty-nine journals were responsible for 7,414 cited articles in zone 2, and 1,409 journals produced 7,422 cited articles in zone 3. CONCLUSIONS: A core collection of veterinary medicine journals would include 49 veterinary medicine journals from zones 1 and 2. Libraries supporting a veterinary curriculum or veterinary research should also include veterinary medical journals from Zone 3, as well as provide access to journals in non-veterinary subjects such as biochemistry, virology, orthopedics, and surgery and a selection of general science and medical journals. PMID- 17082834 TI - "Brimful of STARLITE": toward standards for reporting literature searches. AB - CONTEXT: Systematic reviews of qualitative research studies extend understanding of health care beyond effectiveness to acceptability and user views. OBJECTIVE: The paper surveys reports of qualitative systematic reviews and, by characterizing techniques used to identify articles for inclusion, proposes standards for reporting of literature searches. DATA SOURCES AND STUDY SELECTION: A search of MEDLINE was performed for qualitative systematic reviews published from 1988 to December 2004, supported by searches of CINAHL, Web of Knowledge (including the Science and Social Sciences Citation Index), and the Cochrane Methodology Register, and Internet searches using the Copernic Agent Professional meta-search agent. Studies were included if they used techniques of qualitative synthesis in reviewing research studies in health care. Narrative reviews were excluded. DATA EXTRACTION: Authors, year of publication, sampling strategy, databases, keywords, and other approaches used were extracted. DATA SYNTHESIS: Sixty-four studies were identified, and forty-three met inclusion criteria for this review. A summary of searching methods was produced and used to construct the STARLITE mnemonic (sampling strategy, type of study, approaches, range of years, limits, inclusion and exclusions, terms used, electronic sources). CONCLUSIONS: Considerable variation exists in search methods for qualitative systematic reviews. While diversity in methods is appropriate during the development of review methodology, major concerns remain about the absence of an accepted standard and the consequent poor quality of reporting. PMID- 17082836 TI - An evaluation of five bedside information products using a user-centered, task oriented approach. AB - PURPOSE: The paper compares several bedside information tools using user centered, task-oriented measures to assist those making or supporting purchasing decisions. METHODS: Eighteen potential users were asked to attempt to answer clinical questions using five commercial products (ACP's PIER, DISEASEDEX, FIRSTConsult, InfoRetriever, and UpToDate). Users evaluated each tool for ease-of use and user satisfaction. The average number of questions answered and user satisfaction were measured for each product. RESULTS: Results show no significant differences in user perceptions of content quality. However, user interaction measures (such as screen layout) show a significant preference for the UpToDate product. In addition, users found answers to significantly more questions using UpToDate. CONCLUSION: When evaluating electronic products designed for use at the point of care, the user interaction aspects of a product become as important as more traditional content-based measures of quality. Actual or potential users of such products are appropriately equipped to identify which products rate the highest on these measures. PMID- 17082837 TI - Information-seeking behavior of faculty in one school of public health. PMID- 17082838 TI - Harvesting the best: evidence-based analysis of herbal handbooks for clinicians. PMID- 17082839 TI - Subject specialization in a liaison librarian program. PMID- 17082840 TI - Awareness and attitude of Spanish medical authors to open access publishing and the "author pays" model. PMID- 17082841 TI - Comparison of top-performing search strategies for detecting clinically sound treatment studies and systematic reviews in MEDLINE and EMBASE. PMID- 17082842 TI - Reviving the corporate medical library: using technology to become a cost center. PMID- 17082843 TI - An assignment for building an awareness of the intersection of health literacy and cultural competence skills. PMID- 17082844 TI - Managing database overlap in systematic reviews using Batch Citation Matcher: case studies using Scopus. PMID- 17082845 TI - A blended training approach using videoconferencing for distance education. PMID- 17082846 TI - Sulfonatocalixarenes: molecular capsule and 'Russian doll' arrays to structures mimicking viral geometry. AB - p-Sulfonatocalix[4,5,6,8]arenes are versatile building blocks, able to assemble into 'molecular capsule' arrays based on two calixarenes, as well as a variety of other structural motifs, with the extended structures dominated by the formation of bilayers. For p-sulfonatocalix[4]arene, assembly into nanometre scale spheroids (of either icosahedral or cuboctahedral geometries) as well as nanotubules (all of which take on structural features akin to those of viruses) is possible, depending on the guest molecules and lanthanides present in solution. PMID- 17082847 TI - Luminescent logic function of a surfactant-encapsulated polyoxometalate complex. AB - We have fabricated a novel organic/inorganic hybrid material consisting of multifunctional surfactant-encapsulated polyoxometalloeuropate which functions as a luminescent logic gate with dual output operated by light and metal ion as inputs. PMID- 17082848 TI - Pyridyl thioureas as switchable anion receptors. AB - The binding selectivity of simple pyridyl thioureas in acetonitrile can be completely switched by protonation; hence the neutral thiourea binds acetate, but not chloride or bromide, whereas the protonated thiourea binds strongly to chloride or bromide, but is deprotonated by acetate. PMID- 17082849 TI - Supramolecular porphyrinic prisms: coordinative assembly and solution phase X-ray structural characterization. AB - Supramolecular porphyrin prisms have been obtained via coordinative self-assembly and characterized by 1H NMR, PFG NMR, electronic absorption spectroscopy and synchrotron-based measurements of solution phase X-ray scattering and diffraction. PMID- 17082850 TI - Meso-meso linked core modified 22pi smaragdyrins with unusual absorption properties. AB - The synthesis of the first meso-meso linked core modified smaragdyrins with unusual single photon properties is reported. PMID- 17082851 TI - S-heterocyclic annelated perylene bisimide: synthesis and co-crystal with pyrene. AB - An S-heterocyclic annelated perylene bisimide (PBI) has been prepared from readily available tetrachloro-PBI by a one-step palladium-catalyzed reaction; when co-crystallized with pyrene it gives a unique columnar arrangement of 1 : 2 complexes. PMID- 17082852 TI - An isocyanide probe for heme electronic structure: bis(tert-butylisocyanide) complex of diazaporphyrin showing a unique (dxy)2(dxz, dyz)3 ground state. AB - Isocyanide-bound model hemes always adopt the (dxz, dyz)4(dxy)1 ground state, however, we have found that the replacement of porphyrin by diazaporphyrin leads to the formation of an unprecedented low-spin bis(tert-butylisocyanide) complex with the (dxy)2(dxz, dyz)3 ground state. PMID- 17082853 TI - Synthesis of a [2]rotaxane through first- and second-sphere coordination. AB - In an effort to expand the application of a new template from interpenetrated to interlocked molecular species, we report the synthesis of a new [2]rotaxane by means of both first- and second-sphere coordination of a palladium(II) dichloride subunit. PMID- 17082854 TI - Synthesis of asymmetrically substituted 1,4,7,10-tetraazacyclododecanes for the triggered near infrared emission from lanthanide complexes. AB - A synthetic strategy to prepare asymmetrically substituted 1,4,7,10 tetrazadodecane derivatives was developed to prepare a novel series of photoactive donor-acceptor quencher triads based on Yb and Nd complexes; a nucleoside quencher is used to regulate the extent of energy transfer between the donor and the acceptor. PMID- 17082855 TI - Nickel-catalysed intramolecular alkene insertion into cyclobutanones. AB - A nickel(0) catalyst converted 3-styrylcyclobutanones into benzobicyclo[2.2.2]octenones by an intramolecular insertion of the vinyl moiety into the cyclobutanone skeleton. PMID- 17082856 TI - Synthesis of the mycolactone core by ring-closing metathesis. AB - The undecenolide core of mycolactone was synthesized by ring-closing metathesis and the structure confirmed using single-crystal X-ray diffraction techniques. PMID- 17082857 TI - An unprecedented heterotrimetallic Fe/Cu/Co core for mild and highly efficient catalytic oxidation of cycloalkanes by hydrogen peroxide. AB - An unprecedented hexanuclear heterotrimetallic Fe/Cu/Co complex bearing two Cu(mu O)2Co(mu-O)2Fe cores is easily prepared by self-assembly and acts as a remarkable catalyst for the peroxidative oxidation of cycloalkanes under mild conditions. PMID- 17082858 TI - Photochemical asymmetric synthesis of phenyl-bearing quaternary chiral carbons using chiral-memory effect on beta-hydrogen abstraction by thiocarbonyl group. AB - Quaternary chiral carbons were effectively generated from tertiary chiral carbons via photochemical intramolecular beta-hydrogen abstraction reaction of thioimides involving the highly-controlled chiral-memory effect. PMID- 17082859 TI - Sol-gel encapsulation extends diatom viability and reveals their silica dissolution capability. AB - Several strains of diatom exhibit a long-term viability in silica gels and demonstrate the ability to dissolve the silica in their surroundings. PMID- 17082860 TI - VO(salen)(X) catalysed asymmetric cyanohydrin synthesis: an unexpected influence of the nature of anion X on the catalytic activity. AB - The nature of the anionic ligand X (X = EtOSO3, BF4, Cl, Br, OSO2CF3, F or CN) in vanadium(V)salen complexes [V+ O(salen) X-] was found to have a significant influence on the catalytic activity of the complexes, but not on their enantioselectivities; with the complexes in which X = Cl or F being most active and the complex with X = OSO2CF3 being totally inactive. PMID- 17082861 TI - Direct methane conversion to methanol by ionic liquid-dissolved platinum catalysts. AB - Ternary systems of inorganic Pt salts and oxides, ionic liquids and concentrated sulfuric acid are effective at catalyzing the direct, selective oxidation of methane to methanol and appear to be more water tolerant than the Catalytica reaction. PMID- 17082862 TI - Electrophilic aromatic substituted luciferins as bioluminescent probes for glutathione S-transferase assays. AB - New highly sensitive latent bioluminescent luciferin substrates were designed and synthesized for monitoring mammalian glutathione S-transferase (GST) and Schistosoma japonicum enzyme activities. PMID- 17082863 TI - Coupling ligand recognition to protein folding in an engineered variant of rabbit ileal lipid binding protein. AB - We have engineered a variant of the beta-clam shell protein ILBP which lacks the alpha-helical motif that caps the central binding cavity; the mutant protein is sufficiently destabilised that it is unfolded under physiological conditions, however, it unexpectedly binds its natural bile acid substrates with high affinity forming a native-like beta-sheet rich structure and demonstrating strong thermodynamic coupling between ligand binding and protein folding. PMID- 17082864 TI - Ionic liquid buffers: a new class of chemicals with potential for controlling pH in non-aqueous media. AB - Ionic liquids with buffering characteristics, synthesized by the reaction of [RMIM]OH base moieties with phthalic and tartaric acid, respectively, are potential reagents for controlling pH in non-aqueous media; remarkable [Base]/[Acid] molar ratio dependence of the catalytic activities has been observed in the hydrogenation of olefins with [RuCl2(PPh3)3] complex in DMF and [BMIM][BF4]. PMID- 17082865 TI - Carbon nanotube stabilised emulsions for electrochemical synthesis of porous nanocomposite coatings of poly[3,4-ethylene-dioxythiophene]. AB - The amphiphobic nature of carbon nanotubes allows them to stabilise droplets of water-insoluble monomers dispersed in the aqueous nanotube suspension, leading to a flexible route to electrochemical synthesis of useful nanoporous composites of nanotube doped conducting polymers, and potentially to other chemistry involving reactants incompatible in the same medium. PMID- 17082866 TI - Ultrathin corundum-type In2O3 nanotubes derived from orthorhombic InOOH: synthesis and formation mechanism. AB - Single-crystalline metastable corundum-type In2O3 nanotubes were prepared by annealing solvothermally synthesized InOOH nanotubes under ambient pressure at 300 degrees C, and the formation mechanism of the nanotubes was investigated. PMID- 17082867 TI - Switching catalytic reaction conducted in pore void of mesoporous material by redox gate control. AB - A molecular gate attached on the pore outlet of a mesoporous material, which is opened and closed by redox system of thiol groups, effectively switched the progress of a catalytic reaction promoted by the acidic site in the pore void. PMID- 17082868 TI - Concise synthesis of the (+/-)-Nb-desmethyl-meso-chimonanthine. AB - The first total synthesis of the bis-pyrroloindoline alkaloid (+/-)-Nb-desmethyl meso-chimonanthine, having a pseudo C2-symmetry, was realised in a seven-step convergent sequence without the use of protecting groups. PMID- 17082869 TI - Self-exchange electron transfer in high oxidation state non-oxo metal complexes: amavadin. AB - The electron transfer self-exchange rate constant between the oxidized and reduced forms of amavadin equals approximately 1 x 10(5) dm3 mol(-1) s(-1) at 25 degrees C and represents the first unambiguous example for a vanadium(IV/V) couple. PMID- 17082870 TI - Synthesis and characterization of a trigonal bipyramidal supramolecular cage based upon rhodium and platinum metal centers. AB - The reaction of 4-ethynyl-pyridine with tert-butyl lithium followed by its addition to (Me3tacn)RhCl3 affords the facial octahedral complex (Me3tacn)Rh(CCPy)3, condensation of which with the square planar complex cis (DCPE)Pt(NO3)2 results in a self-assembled trigonal bipyramidal cage with Rh(III) and Pt(II) atoms occupying the vertices. PMID- 17082871 TI - Moral dilemmas in neonatology as experienced by health care practitioners: a qualitative approach. AB - During the last two decades there has been an enormous development in treatment possibilities in the field of neonatology, particularly for (extremely) premature infants. Although there are cross-cultural differences in treatment strategy, an overview of the literature suggests that every country is confronted with moral dilemmas in this area. These concern decisions to initiate or withhold treatment directly at birth and, later on, decisions to withdraw treatment with the possible consequence that the child will die. Given that the neonate cannot express his or her own will, who will decide? And on the basis of what information, values and norms? We explored some of these issues in daily practice by interviewing a small sample of health care practitioners in a Dutch university Neonatal Intensive Care Unit (NICU). It turned out that experiencing moral dilemmas is part of their daily functioning. Nurses underline the suffering of the newborn, whereas physicians stress uncertainty in treatment outcome. To make the best of it, nurses focus on their caring task, whereas physicians hope that future follow-up research will lead to more predictable outcomes. As for their own offspring, part of these professionals would hesitate to bring their own extremely premature newborn to a NICU. For the most oppressing dilemma reported - terminating an already initiated treatment - we propose the concept of 'evidence shift' to clarify the ambiguous position of uncertainty in decision making. PMID- 17082872 TI - Heterosis associated gene expression in maize embryos 6 days after fertilization exhibits additive, dominant and overdominant pattern. AB - Heterosis is important for conventional plant breeding and is intensively used to increase the productivity of crop plants. Genetic processes shortly after fertilization might be of particular importance with respect to heterosis, because coordination of the diverse genomes establishes a basis for future performance of the sporophyte. Here we demonstrate a strong crossbreeding advantage of hybrid maize embryos as early as 6 days after fertilization in a modern maize hybrid and provide the first embryo specific analysis of associated gene expression pattern at this early stage of development. We identified differentially expressed genes between hybrid embryos and the parental genotypes by a combined approach of suppression subtractive hybridization and differential screening by microarray hybridizations. Association of heterosis in embryos with genes related to signal transduction and other regulatory processes was implied by the enrichment of these functional classes among the identified gene set. Quantitative RT-PCR analysis validated the expression pattern of 7 of 12 genes analysed and revealed predominantly additive, but also dominant and overdominant expression patterns in hybrid embryos. These patterns indicate that gene regulatory interactions among parental alleles act at this early developmental stage and the genes identified provide entry points for the exploration of gene regulatory networks associated with the specification of the phenomenon heterosis in the plant life cycle. PMID- 17082873 TI - On the certification of cadmium at trace and ultratrace levels in standard reference materials using ID ICP-MS. AB - Analytical methods used for the isotope dilution inductively coupled plasma mass spectrometric (ID-ICP-MS) measurement of Cd at microg kg(-1) and sub-microg kg( 1) levels are described and applied to the certification of new dietary supplement, blood, and serum Standard Reference Materials (SRMs). The materials are: SRM 3240 Ephedra sinica Stapf Aerial Parts, SRM 3241 Ephedra sinica Stapf Native Extract, SRM 3243 Ephedra-Containing Solid Oral Dosage Form, SRM 3244 Ephedra-Containing Protein Powder, SRM 966 Toxic Metals in Bovine Blood, Level 1 (L1) and Level 2 (L2), and SRM 1598a Animal Serum. The concentration of Cd in the materials ranges from 120 microg kg(-1) down to 0.03 microg kg(-1). At these levels, the factors that most influence the accuracy of the ICP-MS data are the procedure blank and spectral and nonspectral interferences. Nonspectral interference, caused by the high concentration of dissolved solids in the matrices investigated, resulted in signal suppression. Matrix separation was used to enhance signal intensity and to reduce spectral interference for the accurate determination of Cd in SRM 1598a and SRM 3244. Chromatographic separation procedures using Chelex for SRM 1598a and anion exchange for SRM 3244 were optimized to achieve the desired separation characteristics without substantially increasing the procedure blank. Sensitivity for the determination of Cd in serum was additionally enhanced through the use of desolvation nebulization. We determined that separations were not required for the accurate ICP-MS determination of Cd in SRM 3240, SRM 3241, SRM 3243, and SRM 966 L2 under optimized analysis conditions. These samples were diluted to a minimum volume and introduced to the ICP-MS via low flow (40-100 microL/min) microconcentric nebulizers. SRM 966 L1 was also analyzed directly, but results were highly variable. The ID-ICP-MS sample preparation and ratio measurement protocols described here resulted in total expanded uncertainties of less than 1% for the determination of 90.85 microg kg(-1) Cd in SRM 3240, and less than 10% total expanded uncertainty for the determination of 0.0468 microg kg(-1) Cd in SRM 1598a. PMID- 17082874 TI - Toward hybridization assays without PCR using universal nanoamplicons. AB - An innovative scheme for signal amplification using random tetramer-modified gold nanoparticles, termed "nanoamplicons," has been developed for hybridization assay without PCR. Large numbers of nanoamplicons could be integrated onto one target, providing much greater amplification than the larger nanoparticles usually adopted. Using M13mp18 single-strand DNA as a target, this concept is shown to be a feasible approach to detecting 0.17 amol L(-1) DNA without target amplification, based on microgravimetric detection of the adsorption of the probe target-nanoamplicons complex via thiol-gold binding. To our knowledge, this method has a sensitivity that is close to that of PCR and superior to those of nanoparticle-based methods reported previously. Additionally, this novel nanoamplicon could be prepared in the same way and used for all diagnostic tests; such universality would make the nanoamplicons highly advantageous for the generalization and standardization of bioassays, and when applying this new technology in clinical laboratories. PMID- 17082875 TI - Simultaneous determination of uric acid and ascorbic acid at a ferrocenium thioglycollate modified electrode. AB - Self-assembled monolayers (SAMS) of chemisorbed thioglycollate on a gold electrode surface have been used as a base interface for the electrostatic adsorption of ferrocenium ion. Electrochemical impedance spectra (EIS) and cyclic voltammetry (CV) were used to evaluate the electrochemical properties of the supramolecular film. The bare gold electrode failed to distinguish the oxidation peaks of ascorbic acid (AA) and uric acid (UA) in phosphate buffer solution (PBS, pH 7.0), while the ferrocenium-thioglycollate modified electrode could separate them efficiently. In differential pulse voltammetric measurements, the prepared gold electrode could separate AA and UA signals, allowing the simultaneous determination of AA and UA. Under optimal conditions and within the linear range of 1.0 x 10(-6) to 5.0 x 10(-4) M, the detection limits of AA and UA achieved were 2.0 x 10(-7) and 1.0 x 10(-7) M, respectively. The applicability of the prepared electrode was demonstrated by measuring AA and UA in human urine without any pretreatment. PMID- 17082876 TI - Determination of n-octanol-water partition coefficients by hollow-fiber membrane solvent microextraction coupled with HPLC. AB - A novel direct method has been developed for determination of n-octanol-water partition coefficients by hollow-fiber membrane solvent microextraction (HFMSME) combined with high-performance liquid chromatography (HPLC). The compound of interest is dissolved in water with sonication and a hollow fiber containing octanol inside is placed in the sample solution to perform microextraction. After microextraction the concentrations in both the aqueous and n-octanol phases are analyzed by HPLC with UV detection. The method was evaluated with ten reference compounds and shown to be suitable for determination of the partition coefficients of organic compounds accurately, cheaply, simply, and quickly. Previously unknown n-octanol-water partition coefficients have been obtained for other compounds by use of the hollow-fiber membrane solvent-microextraction technique. PMID- 17082877 TI - Quantitative determination of ginsenoside Rh2 in rat biosamples by liquid chromatography electrospray ionization mass spectrometry. AB - Ginsenoside Rh2 is a "hot" natural compound with great potential as a new anti cancer drug based on abundant pharmacological experiments. However, no systemic pharmacokinetic study of Rh2 was reported because current analysis methods could not fully meet the requirements. Thus, we developed a simple LC/MS method with highly improved sensitivities for the determination of Rh2 in rat plasma, bile, urine, feces and most tissues. The tissues and feces were firstly homogenized mechanically using buffer and methanol as the media, respectively. Plasma, bile, urine and tissue homogenates were extracted with diethyl ether for sample preparation. Feces homogenates were directly deproteinized with acetonitrile. The subsequent analysis procedures were performed on a Shimadzu LCMS2010A system (electrospray ionization single quadrupole mass analyzer), with an ODS column (150 mm x 2.0-mm i.d., 5 microm) plus a C18 guard column for separation and ammonium chloride (500 micromol) as mobile phase additive. The proportions of mobile phase were changed timely according to gradient programs. Chlorinated adducts of molecular ions [M + Cl]- of Rh2 at m/z 657.35 and internal standard digitoxin at m/z 799.55 were monitored in selective ion monitoring mode of negative ions. The method was validated to be accurate, precise and rugged with good linearity in all matrices, according to the FDA guidelines. The lower limits of quantitation in rat plasma, urine and feces were 0.2, 0.2 and 20 ng/mL respectively. Stability studies were also performed, indicating that there were no stability-related problems in the analytical procedure of Rh2. The proposed method was successfully applied to the preclinical pharmacokinetic research of Rh2 in rats, including plasma kinetics, tissue distribution and excretion studies. PMID- 17082878 TI - Precision of Raman depolarization and optical attenuation measurements of sound tooth enamel. AB - The demineralization of enamel that is associated with early caries formation affects the optical properties of the enamel. Polarized Raman spectroscopy and optical coherence tomography have been used to detect these changes and potentially offer a means to detect and monitor early caries development. The total optical attenuation coefficient as measured by optical coherence tomography and the polarization anisotropy of the Raman peak arising from the symmetric nu(1) vibration of PO4(3-) at approximately 959 cm(-1) have been demonstrated as being sensitive markers of early caries. This ex vivo study on extracted human teeth demonstrates that these measurements can be made with reasonable precision with concomitantly good repeatability and reproducibility in sound enamel. Such reliability is crucial for these techniques to have a practical clinical value. PMID- 17082879 TI - Fluoroquinolones in soil--risks and challenges. AB - Fluoroquinolones (FQs) are among the most important antibacterial agents used in human and veterinary medicine. Because of the growing practice of adding manure and sewage sludge to agricultural fields these drugs end up in soils, where they can accumulate and have adverse effects on organisms. This paper presents an overview of recent developments in the determination of FQs in solid environmental matrices and describes the risks and challenges (persistence, fate, effects, and remediation) which result from their presence in soil. PMID- 17082880 TI - Integrated approach to the problem of pharmaceutical products in the environment: an overview. PMID- 17082881 TI - Effect of addition of antibiotics and an antioxidant on the stability of tissue reference materials for domoic acid, the amnesic shellfish poison. AB - Five separate reference materials (RMs) were prepared from a mussel (Mytilus edulis) tissue containing domoic acid (DA) from scallop hepatopancreas (Pecten maximus). Homogenates were separately spiked with antibiotics, an antioxidant, or a combination of both. Control materials did not contain any additives and were prepared from lightly cooked and autoclaved mussel tissues. Stability studies were run over a 148-day period at three different temperature conditions: -20 degrees C, +4 degrees C and +40 degrees C. DA contents in all materials were characterised by HPLC-UV. Homogeneities were demonstrated at the beginning of the study, with coefficients of variance of less than 4% (n = 9). DA was stable at 20 degrees C in all materials. The control materials showed significant degradation after two days at +40 degrees C, and after eight days at +4 degrees C. Each of the materials containing additives demonstrated better stability during the initial period of the study. In addition there was no significant degradation in any of the materials with additives stored at +4 degrees C over the duration of the study. The material containing a combination of the antibiotics and the antioxidant displayed the best stability of all the materials. There was no significant reduction in DA concentration at all temperature conditions after eight days, and after 32 days the decrease at +40 degrees C was still <20 %. Following this, a DA laboratory reference material (LRM) was prepared and, based on previous results, spiked with both the antioxidant and antibiotics. A short-term stability study on this material gave similar results to the corresponding material in the additives study. This study shows that combined use of the additives investigated in the preparation of a mussel tissue reference material for DA ensures analyte stability for a period of up to eight days at temperatures of up to +40 degrees C, a condition that is particularly important when shipping test materials globally. Aliquots of individual feasibility materials used in the study. PMID- 17082882 TI - Drug-protein interaction with Vpu from HIV-1: proposing binding sites for amiloride and one of its derivatives. AB - Vpu is an 81-amino-acid auxiliary protein of the genome of HIV-1. It is proposed that one of its roles is to enhance particle release by self-assembling to form water-filled channels enabling the flux of ions at the site of the plasma membrane of the infected cell. Hexamethylene amiloride has been shown to block Vpu channel activity when the protein is reconstituted into lipid bilayers. In a docking approach with monomeric, pentameric and hexameric bundle models of Vpu corresponding to the transmembrane part of the protein, a putative binding site of hexamethylene amiloride is proposed and is compared with the site for the nonpotent amiloride. The binding mode for both ligands is achieved by optimizing hydrogen bond interactions with serines. Binding energies and binding constants are the lowest for protonated hexamethylene amiloride in the pentameric bundle. PMID- 17082883 TI - Analysis of subcellular surface structure, function and dynamics. AB - Analytics of single biological cells allows quantitative investigation from a structural, functional and dynamical point of view and opens novel possibilities to an unamplified subcellular analysis. In this article, we report on three different experimental methods and their applications to single cellular systems with a subcellular sensitivity down to the single molecule level. First, the subcellular surface structure of living bacteria (Corynebacterium glutamicum) was investigated with atomic force microscopy (AFM) at the resolution of individual surface layer (S-layer) proteins; discrimination of bacterial strains that lack the expression of hexagonally packed surface layer proteins was possible. Second, quantitative measurement of individual recognition events of membrane-bound receptors on living B-cells was achieved in single cell manipulation and probing experiments with optical tweezers (OT) force spectroscopy. And third, intracellular dynamics of translocating photoactivatable GFP in plant protoplasts (Nicotiana tabacum BY-2) was quantitatively monitored by two-photon laser scanning microscopy (2PLSM). PMID- 17082884 TI - [Neuromuscular blockades. Agents, monitoring and antagonism]. AB - Currently, the main aims of using neuromuscular blocking agents during general anaesthesia are the improvement of surgical and intubation conditions. Neuromuscular blocking agents themselves are neither analgesic nor anaesthetic. All agents interact with the acetylcholinergic receptor at the neuromuscular junction and induce a blockade either through a continuous activation imitating the effect of acetylcholine or through a competitive antagonism against acetylcholine. Succinylcholine is the only depolarizing muscle relaxant that is in clinical use. Non-depolarizing neuromuscular blockers may be grouped by their chemical structure into benzylisoquinolines or aminosteroids, and cover the complete range from short and intermediate, to long acting agents. Possible adverse drug reactions to the single agents are also related to their mechanism of action. Moreover, pharmacokinetic properties and effects such as histamine liberation could play an important role when choosing a myorelaxant. The depth of a neuromuscular block and recovery from paralysis can be monitored using qualitative and quantitative techniques. Therefore, the monitoring of neuromuscular recovery plays an important role in the prevention of postoperative complications due to residual paralysis. In case of residual paralysis, cholinesterase inhibitors are suitable for reversal. PMID- 17082885 TI - Preliminary study on eye colour in Japanese macaques (Macaca fuscata) in their natural habitat. AB - Eye colour in Japanese macaques shows apparent differences between individuals, continuously ranging from orange (bright), through shades of yellow and hazel blue to dark blue (dark). We arbitrarily classified them into either 'yellow' eyes or 'blue' eyes based on the yellow area occupying in the iris' peripupillary ring. Most Japanese macaques have yellow eyes after infant phase, whilst 19, 17, 12, and 15% of monkeys (>6 months, sexes combined) have blue-eye in studied two groups of Shodoshima and two groups of Takasakiyama, respectively. Frequency of eye colour did not differ between males and females, but significantly differed in each age class. Blue eyes significantly more frequently occurred in newborns, infants and aged monkeys than in juveniles and prime adults. Data from mother infant pairs indicated eye colour could be inherited from their parents. A case of asymmetric eye colour in Japanese macaques was found from a sample of 1962 individuals. Eye colour variation of Japanese macaques was discussed in relation to those of humans and rhesus macaques. A possible evolutionary model of eye colour in Japanese macaques was discussed. PMID- 17082886 TI - Effects of parity and age on female attraction to faces of infants and neonates in rhesus macaques. AB - This study investigated the effects of parity and age on female rhesus macaque attention toward infants, and assessed whether the faces of neonates are more attractive than those of older infants. Six nulliparous and six multiparous females were shown digitized images of neonates' and 5- to 6-month-old infants' faces. Attention and preferences for images were measured by gaze duration and other picture-directed behaviors, including lip smacking, approaches, and presentations. As predicted, nulliparous females displayed significantly longer gaze durations for images than did multiparous females. There were no significant differences in gaze duration for faces of neonates and those of infants, but images of infants were approached more frequently than images of neonates. This difference is tentatively explained on the basis of differences in female familiarity with neonates' and infants' faces and differences in opportunities for allomothering with neonates and infants. PMID- 17082887 TI - Toxicity and efficacy of protracted low dose temozolomide for the treatment of low grade gliomas. AB - Protracted low dose temozolomide (75 mg/m(2)/day on days 1-21 of 28 days) offers potential advantages over standard temozolomide schedules (200 mg/m(2)/day on days 1-5 of 28 days) including greater cumulative drug exposure and depletion of O(6)-alkylguanine DNA alkyltransferase levels, theoretically overcoming intrinsic chemoresistance. We retrospectively review our experience in 25 patients with pathologically proven low grade gliomas (LGG) treated with protracted low dose temozolomide to primarily quantify its toxicity and secondarily to assess efficacy. None had previously received radiation. Tumor response was graded based on changes in tumor size, steroid requirements, and clinical exam. About 243 cycles of protracted low dose temozolomide were administered. Three patients (12%) were changed to standard temozolomide dosing due to side effects, including intractable nausea (n = 2) and multiple cytopenias (n = 1). The most frequent toxicities were fatigue (76%), lymphopenia (72% [48% high grade]), constipation (56%), and nausea (52%). High grade toxicities (other than lymphopenia) included secondary malignancy, pruritus, hyponatremia, neutropenia, leukopenia, and cognitive decline (n = 1 for each). Tumor response rate was 52% and and disease control rate was 84%. Six month PFS was 92% and 12 month PFS was 72%. Response rates and PFS were independent of pathological subtype, deletion status, and indication for chemotherapy. Protracted low dose temozolomide has a distinct spectrum of toxicities compared to standard dosing but is well tolerated in most patients and may provide improved response rates compared to standard dosing. The results of larger randomized trials are needed to assess its potential advantages over other management schemes. PMID- 17082888 TI - The GH1/IGF-1 axis polymorphisms and their impact on breast cancer development. AB - The growth hormone 1/insulin-like growth factor-1 (GH1/IGF-1) axis plays an essential role in the development of the breast by regulating cell proliferation, differentiation and apoptosis. Imbalances within this axis lead to an aberrant signalling and recent research has focussed on the overexpression of these growth factors and their involvement in breast cancer development. The increased understanding of the molecular mechanisms and signalling pathways connected to the GH1/IGF-1 axis has provided important insights into aetiology, prevention and therapy for breast cancer. However, to identify the contribution of the GH1/IGF-1 signalling pathway to cancer risk still remains a challenge since the results of various studies are controversial. Here, we discuss the influence of low penetrance polymorphisms in the genes along the GH1/IGF-1 axis and their impact on hormone levels and cancer risk, especially breast cancer. We point out what is known about the effects of the variants and show how the interaction of genetic variants affects breast cancer risk. PMID- 17082890 TI - Mesenteric desmoid tumors in Singapore familial adenomatous polyposis patients: clinical course and genetic profile in a predominantly Chinese population. AB - PURPOSE: This study examined the mutational profile of the adenomatous polyposis coli gene in relation to the development of desmoid tumors in familial adenomatous polyposis patients from a predominantly Chinese population. METHODS: This is a retrospective review of all patients with familial adenomatous polyposis coli from the Singapore Polyposis Registry. Identification of specific adenomatous polyposis coli gene mutation was performed and clinical course of associated desmoid disease obtained from case records and a computerized database. RESULTS: Two hundred five patients from 75 families afflicted with familial adenomatous polyposis coli were reviewed, with gene mutations identified in 107 patients. Of these, 23 (11.2 percent) developed desmoids. The male-to female ratio was 1:1.3 and the ethnic distribution was Chinese (n=17) and Malay (n=6). Of the 92 patients with mutations 5' to codon 1444, 11 patients (12 percent) developed desmoids compared with 6 of 15 (40 percent) patients with adenomatous polyposis coli gene mutations 3' to codon 1444 (P<0.01). The clinical course of desmoid tumors can be divided into stable (n=11), variable (n=3), progressive (n=6), and aggressive growth (n=3). Only 3 (13 percent) patients with aggressive tumor growth required chemotherapy. There was no correlation between the site of mutation and the clinical progression of the desmoids. Seventy-four percent of these desmoids (17/23) developed at a mean interval of 2.98 years after restorative proctocolectomy, while only 30 percent (7/23) were diagnosed preoperatively or discovered during the initial surgery. The most common complications related to the mesenteric desmoids were intestinal obstruction (21.7 percent), ureteric obstruction (17.4 percent), and encasement of superior mesenteric vessels (13 percent). CONCLUSION: The clinical course of desmoids in an individual familial adenomatous polyposis patient remains unpredictable and no reliable genetic marker is available for prognostication in desmoid disease. PMID- 17082891 TI - Efficacy of anal fistula plug in closure of cryptoglandular fistulas: long-term follow-up. AB - PURPOSE: The long-term efficacy of Surgisis anal fistula plug in closure of cryptoglandular anorectal fistulas was studied. METHODS: Patients with high cryptoglandular anorectal fistulas were prospectively studied. Additional variables recorded were: number of fistula tracts, and presence of setons. Under general anesthesia and in prone jackknife position, patients underwent irrigation of the fistula tract by using hydrogen peroxide. Each primary opening was occluded by using a Surgisis anal fistula plug, which was securely sutured in place at the primary opening and tacked to the periphery of the secondary opening. RESULTS: Forty-six patients were prospectively enrolled during a two year period. Follow-up was six months to two years (median, 12 months). At final follow-up, all fistula tracts had been successfully closed in 38 patients, for an overall success rate of 83 percent. Seven patients had multiple tracts, for a total of 55 fistula tracts in the series. Of the 55 individual tracts, 47 (85 percent) were closed at final follow-up. Patients with one primary opening were most likely to have successful closure by using the anal fistula plug, although this was not significant. Successful closure was not correlated with the presence of setons. CONCLUSIONS: Long-term closure of cryptoglandular anorectal fistula tracts using Surgisis anal fistula plug is safe and successful in 83 percent of patients and 85 percent of tracts. PMID- 17082892 TI - Inflammatory polyps: a cause of late bleeding in stapled hemorrhoidectomy. AB - PURPOSE: Stapled hemorrhoidectomy has become a popular treatment of hemorrhoids, mainly because of the benefits of greatly reduced postoperative pain. However, complications unique to the new procedure have been reported. This study was designed to review our series of 82 patients with a focus on complications, with particular focus on late bleeding caused by inflammatory polyps at the staple line. METHODS: A review of 82 patients who underwent stapled hemorrhoidectomy was conducted. Indications included bleeding in 29 patients, prolapse in 19, combined bleeding and prolapse in 31, and pain in 3. The procedure was performed in an outpatient setting with the Ethicon ILS 33 stapler in the standard fashion. RESULTS: Late bleeding as a result of inflammatory polyps was encountered in nine patients (11 percent). Bleeding was mild, and all cases resolved after excision of the polyps. Other complications occurred with the following frequency: urinary retention (4.9 percent), submucosal hematoma (2.4 percent), early bleeding (1.2 percent), recurrent hemorrhoids (2.4 percent), thrombosed external hemorrhoids (2.4 percent), and fecal urgency (3.7 percent). CONCLUSIONS: Bleeding from inflammatory polyps occurs in a significant number of patients undergoing stapled hemorrhoidectomy. Mild bleeding several weeks or months after the procedure should prompt a search for inflammatory polyps at the staple line. Simple excision of the polyps was adequate treatment and has not resulted in rebleeding. PMID- 17082893 TI - Blastomyces dermatitidis of the perianal skin: report of a case. AB - Isolated fungal infections of the perianal skin are rare and their diagnosis is frequently overlooked. We report a case of a 78-year-old male patient who presented with a friable, violaceous, papulopustular lesion, with heaped-up edges along the anal verge. Biopsy revealed unicellular yeast consistent with blastomycosis. The patient was treated with itraconazole with resolution of this lesion. An extensive MEDLINE literature review from 1958 to the present indicates that this is an uncommon manifestation of cutaneous blastomycosis. A summary of the medical literature is presented with a review of the characteristics, diagnosis, and management of blastomycosis. PMID- 17082894 TI - Pseudoaneurysm of the inferior gluteal artery: an unusual complication after abdominoperineal resection for rectal cancer. Report of a case. AB - We report an unusual case of profuse, sporadic hemorrhage from a chronically infected perineal wound after an abdominoperineal resection. Surgical exploration and angiography on two occasions failed to identify the source of bleeding. A pseudoaneurysm of the internal iliac artery was eventually identified with a provocative angiogram using tissue plasminogen activator. This aneurysm was successfully treated with embolization with coils. PMID- 17082895 TI - A new retractor system for thoracoscopic thymectomy using the anterior chest wall lifting method. PMID- 17082896 TI - Detection of major depression in Ugandan primary health care settings using simple questions from a subjective well-being (SWB) subscale. AB - OBJECTIVE: To explore whether the 4-item subjective well-being subscale could be used to detect a major depressive illness. Secondly, to describe the prevalence and characteristics of depressed health care attendees at primary healthcare centres. METHOD: Using a descriptive, cross-sectional study design, we interviewed 199 consecutive patients about their socio-demographics, subjective well-being (SWB), major depressive illness symptoms and depression severity. The instruments used were translated into Luganda. RESULTS: Point prevalence of a current Major Depressive Episode (MDE) was 31.6%. Using a one week reference period, we found that experiencing a lot of distress, having less energy or poor health, having poor emotional and psychological adjustment and not being satisfied with life were significantly more common among patients with a current MDE. The 4-item SWB subscale detected depression of up to 87.1% (95% CI: 0.818 0.923). In logistic regression, all four SWB items predicted a current MDE. CONCLUSION: Major depressive illness is a common at primary healthcare level in Uganda. Four simple questions reflecting SWB items have potential to detect diagnosable patients likely to have a current MDE, making general screening procedures less necessary. PMID- 17082897 TI - The subjective consequences of suffering a first episode psychosis: trauma and suicide behaviour. AB - BACKGROUND: The subjective impact of a psychotic breakdown can be profound, potentially resulting in loss of social roles, hopes and aspirations and leading to stigmatisation, trauma and elevated suicide risk. This study aimed to assess the subjective effect and consequences of suffering a first episode of psychosis. It was hypothesised that suicide behaviour would be associated with the negative consequences of psychosis and co-morbid symptomatic-PTSD. METHODS: Patients were assessed by means of a semi-structured interview on their reactions and experience of their psychotic episode and its treatment and by means of standardised methods for psychotic (PANSS) and trauma-related (CAPS) symptoms. RESULTS: A total of 35 patients suffering their first episode of psychosis were interviewed. As a result of the onset of their illness, 77% indicated they had suffered loss or disruption to their life, 60% had thwarted future aspirations, 38% had suffered violence or harassment, 53% had suffered stigma and 50% social exclusion. Totally, 80% felt they had been traumatised by their treatment and 38% were cases for symptomatic-PTSD. Symptomatic-PTSD was significantly associated with involuntary hospitalisation but not psychotic symptoms. Positive psychotic symptoms were associated with harassment, stigma and social exclusion. Suicidal ideation was reported by 40% and 31% reported attempting suicide. Suicidal behaviour was greater in those suffering symptomatic-PTSD but this was not significant, suicidal behaviour was significantly associated with the experience of trauma, but not the severity of that trauma, prior to the onset of their psychosis. CONCLUSIONS: The negative consequences of a psychotic episode are significant. The potential iatrogenic effect of psychiatric care needs to be considered. Interventions need to be developed to reduce traumatisation and suicide risk. PMID- 17082898 TI - Clinically non-functioning pituitary adenoma. AB - Non-functioning pituitary tumors are relatively common. A large number of these tumors are incidentally found pituitary microadenomas (<1 cm) and are usually of no clinical importance. Those tumors that require treatment are generally macroadenomas and come to medical attention because of mass effect and/or hypopituitarism. Visual field defects are present in roughly 70% of patients with non-functioning macroadenoma at the time of diagnosis and the majority of these patients have at least growth deficiency and hypogonadism. By immunocytochemistry, the large majority of these tumors are glycoprotein producing and less commonly they are non-functioning somatotroph, lactotroph or corticotoph adenomas. In contrast to the immunocytochemistry results, only a minority of these tumors actively secrete intact gonadotrophs or glycoprotein subunits. Therapy is directed at eliminating mass effect and correcting hypopituitarism. There are anecdotal reports of tumor shrinkage during therapy with either dopamine agonists or somatostatin agonists; however tumor response to medical treatment is not reliable. For most patients, transphenoidal resection of the tumor is the preferable primary treatment. Surgery improves visual defects in the majority of patients and a lesser number will recover pituitary function. In the past, pituitary radiation was commonly administered following pituitary surgery; however the need for routine radiation has recently been reevaluated. Although tumor recurrence at 10 years post surgery may be as high as 50%, few patients with recurrence will have clinical symptoms. Close follow-up with surveillance pituitary scans should be performed after surgery and radiation therapy reserved for patients having significant tumor recurrence. PMID- 17082899 TI - Response to influenza vaccine in people with non-protective HI antibody titers. AB - The purpose of the study was to determine which factors correlate directly with response to vaccination in such a group of subjects with non-protective HI antibody titers before vaccination. Two vaccines were used, a subunit virus vaccine adjuvanted with MF59 and a split virus vaccine. The analysis indicated that immunization with vaccine adjuvanted with MF59 was an independent variable for immune response against A/H3N2 (OR: 3.51; 95% CI: 1.81-6.79) and B (OR: 2.31; 95% CI: 1.37-3.89). The results suggest that antibody response to vaccine is satisfactory in elderly people previously lacking a protective antibody titer, and that the adjuvanted vaccine reveals a better immunogenicity. PMID- 17082900 TI - Exposure to famine during gestation, size at birth, and blood pressure at age 59 y: evidence from the Dutch Famine. AB - We compared blood pressure of individuals (mean age 59 y) born in western Holland between January 1945 and March 1946 (mothers exposed to the Dutch Famine before or during gestation; n = 359) to blood pressure of unexposed individuals born before or conceived after the famine (n = 299) or same-sex siblings of subjects in series 1 or 2 (n = 313). Mean (SD) systolic and diastolic blood pressure were 140.3 (20.3) and 85.8 (11.0) mmHg, respectively; prevalence of hypertension (prior diagnosis of hypertension or with measured systolic/diastolic blood pressure above 140/90 mmHg) was 61.8%. Birth weight was inversely related to systolic (-4.14 mmHg per kg; 95% confidence interval (CI) -7.24, -1.03; p < 0.01) and diastolic (-2.09 mmHg per kg; 95% CI -3.77, -0.41; p < 0.05) blood pressure and to the prevalence of hypertension (odds ratio 0.67 per kg, 95% CI: 0.49, 0.93) (all age- and sex-adjusted). Any famine exposure of at least 10 weeks duration was associated with elevated systolic (2.77 mmHg; 95% CI 0.25, 5.30; p < 0.05) and diastolic (1.27 mmHg; 95% CI -0.13, 2.66; p = 0.08) blood pressure and with hypertension prevalence (odds ratio 1.44; 95% CI 1.04, 2.00; p < 0.05) in age- and sex-adjusted models. Exposure to famine during gestation may predispose to the development of hypertension in middle age. PMID- 17082901 TI - A comparative study of cells in inflammation, EAE and MS using biomedical literature data mining. AB - Biomedical literature and database annotations, available in electronic forms, contain a vast amount of knowledge resulting from global research. Users, attempting to utilize the current state-of-the-art research results are frequently overwhelmed by the volume of such information, making it difficult and time-consuming to locate the relevant knowledge. Literature mining, data mining, and domain specific knowledge integration techniques can be effectively used to provide a user-centric view of the information in a real-world biological problem setting. Bioinformatics tools that are based on real-world problems can provide varying levels of information content, bridging the gap between biomedical and bioinformatics research. We have developed a user-centric bioinformatics research tool, called BioMap, that can provide a customized, adaptive view of the information and knowledge space. BioMap was validated by using inflammatory diseases as a problem domain to identify and elucidate the associations among cells and cellular components involved in multiple sclerosis (MS) and its animal model, experimental allergic encephalomyelitis (EAE). The BioMap system was able to demonstrate the associations between cells directly excavated from biomedical literature for inflammation, EAE and MS. These association graphs followed the scale-free network behavior (average gamma = 2.1) that are commonly found in biological networks. PMID- 17082902 TI - Identification and characterization of two rice autophagy associated genes, OsAtg8 and OsAtg4. AB - Autophagy is an intracellular process for vacuolar degradation of cytoplasmic components. The molecular machinery responsible for yeast and mammalian autophagy has begun to be elucidated at the cellular level. A genome-wide search revealed significant conservation among autophagy genes in yeast and Arabidopsis. Up till now, however, there is no report about rice autophagy associated genes. Here we cloned OsAtg8 and OsAtg4 from Oryza sativa and detected their expression patterns in various tissues. Immunoblotting analysis showed that carboxyl terminus of OsAtg8 can be cleaved in yeast cell. Mutation analysis revealed that the conserved Gly117 residue of OsAtg8 was essential for its characteristic C terminal cleavage as similar to that found in mammalian and yeast Atg8. We further proved that OsAtg8 interacted with OsAtg4, and this interaction was not affected by the conserved Gly117 mutation. Our results demonstrate that Atg8 conjugation pathway is conserved in rice and may play important roles in rice autophagy. PMID- 17082903 TI - Expansion of symmetric exon-bordering domains does not explain evolution of lineage specific genes in mammals. AB - In order to examine the evolution of lineage specific genes, we analyzed intron phase distributions and exon-bordering domains in primate and rodent specific genes. We found that the expansion of symmetric exon-bordering domains could not explain the evolution of lineage specific genes. Rather internal intron loss of a domain can partially explain the excess of class 1-1 intron phases in the lineage specific genes. We suggest the event that led to excess of symmetric exons in lineage specific genes had little bearing on shaping the phenotypes specific to the individual lineage. Instead, Kruppel-associated box (KRAB) proteins associated with zinc finger C2H2 (zf-C2H2) type are likely to be responsible for the lineage specific function. PMID- 17082904 TI - Genetic diversity and variety composition of cassava on small-scale farms in Uganda: an interdisciplinary study using genetic markers and farmer interviews. AB - Cassava is a tropical crop and grown for its tuberous starchy roots. In Africa it is mainly cultivated by small-scale farmers who observe, select and name their cassava varieties based on morphology, food, social and economic interest. Here we have used an interdisciplinary approach involving farmer interviews, genetic markers and morphological descriptors to study the composition of cassava varieties on small-scale farms in 11 villages located in three districts in Uganda, the genetic structure within and between these varieties and their morphology. The composition of local, newly introduced and improved varieties differed widely between villages and districts. The Ugandan farmers in our study seemed to adopt improved varieties to a greater extent when there was a nearby market, prevalence of disease epidemics and good extension service. We found considerable genetic variation both within and between cassava varieties though the variation was larger between varieties. However, most local and improved varieties showed predominating genotypes at many loci. Accessions of commonly grown varieties meeting farmers' preferences could therefore be selected and implemented in future breeding programmes involving development, dissemination and adoption. The like-named varieties in different villages were genetically similar, demonstrating farmers' ability to differentiate and maintain the same variety over large areas. However, some varieties with different names in different villages showed both genetic and morphological similarity, suggesting that farmers may rename plants when they are introduced into their fields. The large differences found in variety and genetic composition between villages and districts in Uganda may be a result of the diverse needs and growing conditions characteristic for traditional farming system. This suggests that efforts to conserve and increase the genetic diversity in farmers' fields will require policies tailored to each area. PMID- 17082905 TI - Lichens as bioindicators of atmospheric heavy metal pollution in Singapore. AB - Lichens have been used as bioindicators in various atmospheric pollution assessments in several countries. This study presents the first data on levels of heavy metals (As, Cd, Cu, Ni, Pb, and Zn) in lichens at different locations in Singapore, Southeast Asia. Singapore is a fully industrialised island nation, with a prevailing tropical climate and a population of 4 million people within a confined land area of less than 700 km2. The ubiquitous lichen species, Dirinaria picta was collected from six sample sites across Singapore and analysed for heavy metals using inductively coupled plasma mass spectrometry (ICPMS). No significant relationship existed between metal levels in lichen and soil, indicating that accumulated metals in lichen are primarily derived from the atmosphere. Peak concentrations of zinc (83.55 microg g(-1)), copper (45.13 microg g(-1)) and lead (16.59 microg g(-1)) in lichens were found at Sembawang, Jurong and the National University of Singapore campus which are locations associated with heavy petroleum and shipping industries, and road traffic respectively. The mean heavy metal levels of lichen samples in Singapore were found to be at the upper range of values reported in the literature for temperate countries. PMID- 17082906 TI - Environmental impact of the marine aquaculture in Gulluk Bay, Turkey. AB - The effects of marine aquaculture on the environment were evaluated by studying the water quality of Gulluk Bay (Turkey). Marine aquaculture, both extensive and intensive, is one of the most important activities carried out in this area. The intensive culture of fish (Dicentrarchus labrax) is the most important polluting element in Gulluk Bay. Records of long term (seasonally) monitoring of the ambient water dissolved oxygen concentration, the inorganic nutrients (ammonia, nitrate, nitrite, and phosphate) and chlorophyll a concentrations were generated to assess sea water quality of Gulluk Bay. Surface water (0.5 m) samples were collected from seven fish farm areas in the cage of Gulluk Bay (here after reported as the cage stations). Reference surface water was also concurrently sampled at three stations. Modifications in the cage stations water quality were assessed as the difference between the magnitude of a specific parameter recorded at cage station and the concurrently recorded value of the parameter at the reference station, relative to the mean value at the reference station. In this study, results in the specific modifications are discussed at the study period. An aquaculture area had consistently positive modifications in phosphate concentration, but not in nitrogen or chlorophyll a concentrations. Chlorophyll a concentrations showed non positive modifications only in June. In addition to these site specific modifications nitrogen concentrations were positively modified in August and October. PMID- 17082907 TI - Significance of radical cystectomy for bladder cancer in patients over 80 years old. AB - OBJECTIVES: The objective of this study was to evaluate the usefulness of radical cystectomy for bladder cancer in elderly patients. MATERIALS AND METHODS: This study included 72 patients aged > or =80 years (group A) who underwent radical cystectomy and urinary diversion between January 1995 and December 2003, and the clinical outcome of these patients were compared with those of 557 patients aged <80 years (group B) undergoing radical cystectomy during the same period as group A. RESULTS: As the procedure for urinary diversion, ureterocutaneostomy was most frequently performed in group A (87.5%), while neobladder creation was most common in group B (43.8%). Despite the absence of significant differences in tumor grade and incidence of lymph node metastasis between these two groups, pathological stage in group A was significantly greater than that in group B. The perioperative mortality rate in group A was significantly higher than that in group B, whereas the incidences of both early and late postoperative complications in group A were similar to those in group B. Cancer-specific survival in group A was significantly lower than that in group B; however, among patients with disease < or =pT2, there was no significant difference in cancer specific survival between these two groups. CONCLUSIONS: These findings suggest that an aggressive surgical approach may be an optimal therapeutic strategy for properly selected elderly patients who require definitive therapy for locally invasive bladder cancer, particularly in those with disease < or =pT2. PMID- 17082908 TI - Specific expression of annexin A8 in adult murine stratified epithelia. AB - Annexin A8 is a member of the annexin family of calcium-regulated membrane binding proteins. In this report, we investigated the expression of annexin A8 in adult mouse organs. Northern blot analysis of adult mouse organs showed that a single annexin A8 transcript of 1.9 kb is expressed most strongly in skin, eye and tongue. In situ hybridisations using annexin A8-specific probes revealed that in the stratified epithelia of the tongue and the early postnatal epidermis, annexin A8 transcription could be detected in basal and suprabasal layers of these stratified epithelia. Western blot analyses using a murine ANXA8-specific antiserum showed, that the 36 kD ANXA8 protein was most abundant in the skin and tongue. The abundance of ANXA8 protein in the skin increased during postnatal days 1-18 and was immunohistochemically localised in suprabasal layers of the epidermis. In the tongue epithelium as well, ANXA8 protein was found in suprabasal layers. ANXA8 immunoreactivity was also found in suprabasal layers of the stratified epithelia of the oesophagus and the forestomach, while it was detected in all layers of the cornea epithelium and in the cornea endothelium of the eye. We also investigated the expression of retinoic acid receptor alpha protein (RARA) and ANXA8 in the epidermis immunohistochemically. While RARA immunoreactivity was exclusively detected in the basal layer, ANXA8 immunoreactivity was restricted to suprabasal layers of the epidermis. Thus, ANXA8 protein is most abundant in stratified epithelia of the postnatal mouse. Its location in the suprabasal layers suggests that ANXA8 may be associated with the terminal differentiation of epithelial cells in these tissues. PMID- 17082909 TI - Dehydroepiandrosterone sulphate enhances IgG and Interferon-gamma production during immunization to tuberculosis in young but not aged mice. AB - Ageing of the endocrine system (endocrinosenescence) has been closely related to immunosenescence. Dehydroepiandrosterone sulphate (DHEAS), a steroid hormone produced by the adrenals with reported enhancing immunomodulatory properties, consistently decline during ageing in parallel to detrimental increase in peripheral glucocorticoids. We investigated here the adjuvant effects of DHEAS during intraperitoneal immunization to Mycobacterium tuberculosis heat shock protein 70 (mycHSP70) in old (24 months) as well as young (3 months) BALB/c mice. Both young and old mice had significantly higher Immunoglobulin G (IgG) levels following immunization. Young mice co-immunized with mycHSP70-DHEAS presented an early increase in specific IgG levels and showed increased Interferon-gamma production compared to old mice. Also, T cells of immunized young animals were consistently more resistant to the immunosuppressive effects of glucocorticoids and to DHEAS. DHEAS was not effective in modulating antigen-specific T-cell proliferation, Interleukin-2 production or percentage of recent activated T-cell subsets (CD4 + CD69 + and CD8 + CD69 +). Our data further indicate mycHSP70 as a putative good antigen in vaccine to tuberculosis. Our data also suggest that DHEAS produced adjuvant effects upon humoral and some cellular immune responses of young, but not old mice and indicate that immunization with DHEAS is capable of changing T-cell responses to steroids. PMID- 17082910 TI - Survival and longevity improvements at extreme ages: an interpretation assuming an ecological stress theory of aging. AB - The primary determinant of survival during aging is the energetic efficiency and metabolic stability required to counter the accumulated internal and external stresses of a lifetime. Hence, genetically stress-resistant individuals should accumulate with age; frailer, less robust, less energetically efficient and less metabolically stable individuals should succumb in parallel. This selection process implies the accumulation of energetically efficient stress-resistant individuals with age to the exclusion of all others. High additive genetic variability for survival is expected under extreme circumstances, however there is limited evidence close to the absolute extremes of life that diversity may fall. At this stage, only a few highly adaptive, oxidative-stress-resistant and presumably somewhat homozygous genotypes should remain. Therefore a fall in variability may occur in these outliers, when frailer individuals are unable to cope and are eliminated at extreme ages. This process could provide an explanation of mortality-rate declines in domesticated (laboratory) and free living populations of the extremely old. That is, mortality-rate declines may be an expectation from a process of genetic sorting resulting from the accumulated responses to environmental stress over time. Application of an ecological stress theory of aging, which combines the external stresses to which organisms are exposed with internal stresses, appears to be the prerequisite for this conclusion. PMID- 17082911 TI - Catalase effect on cell death for the improvement of recombinant protein production in baculovirus-insect cell system. AB - Baculovirus expression vector system (BEVS) in host insect cells is a powerful technology to produce recombinant proteins, as well as virus-like particles (VLP). However, BEVS is based on baculovirus infection, which limits the recombinant protein production by inducing insect cell death. Herein a new strategy to enhance cell life span and to increase recombinant protein production was developed. As baculovirus infection induces cellular oxidative stress, the ability of several antioxidants to inhibit cell death was tested during infection. The production of rotavirus structural proteins was used as model to analyse this new strategy. We found that only catalase is able to partially prevent cell death triggered by baculovirus infection and to inhibit lipid peroxidation. An increase in recombinant protein production was coupled with the partial cell death inhibition. In summary, the addition of catalase is a promising strategy to improve recombinant protein production in BEVS, by delaying insect cell death. PMID- 17082912 TI - Feeding strategies for E. coli fermentations demanding an enriched environment. AB - The addition of a carbon nutrient feed to a fed-batch cultivation is often not enough to obtain satisfactory growth and/or production. In some cases, an additional feed with for example supplementary amino acids or complex media is required. This work presents the development of feeding strategies where more than one feed is required and the knowledge of the growth requirements is low. Simulations and cultivations with E. coli are shown using the proposed feed controllers which are based on a probing control concept. The strategies work well and they can be used to shorten the process development phase considerably. PMID- 17082913 TI - Predictive controller evaluation including non-stationary high frequency noise and outliers for batch solid substrate fermentation bioreactors. AB - Optimum operation and automatic control of large-scale solid substrate fermentation (SSF) bioreactors is difficult. Though advanced control algorithms can handle most challenges encountered properly, for real-time SSF processes such controllers are expensive and time consuming to design and tune. With these considerations, advanced control algorithm tests using realistic simulations appear more appropriate. We used a phenomenological process model of an SSF pilot bioreactor, coupled with a realistic noise model, to test linear model predictive controllers. We focused on the effect noise has on the performance of the control algorithms, and how to enhance performance using a combination of low-pass (Butterworth) and outlier shaving (Hampel) filters. In simulations undertaken directly with the phenomenological model it was relatively straightforward to achieve good control performance. Nevertheless, control degraded sharply when the output of the phenomenological model was contaminated with noise using our realistic noise model, even with proper signal filtering. PMID- 17082914 TI - Oxygen transfer to slurries treated in a rotating drum operated at atmospheric pressure. AB - The objective of this work was to determine (1) the effect of rotational speed (N) and lifters on the oxygen transfer coefficient (k (L)) of a mineral solution and (2) the effect of solids concentration of a slurry soil-mineral solution on k (L), at a fixed value N (0.25 s(-1)); in both cases the treatment was carried out in an aerated rotating drum reactor (RDR) operated at atmospheric pressure. First, the k (L) for the mineral solution was in the range 6.38 x 10(-4)-7.69 x 10(-4) m s(-1), which was of the same order of magnitude as those calculated for closed rotating drums supplied with air flow. In general, k (L) of RDR implemented with lifters was superior or equal to that of RDR without lifters. For RDR implemented with lifters, k (L) increased with N in the range 6.65 x 10( 4)-10.51 x 10(-4) m s(-1), whereas k (L) of RDR without lifters first increased with N up to N = 0.102 s(-1), and decreased beyond this point. Second, regarding soil slurry experiments, an abrupt fall of k (L) (ca. 50%) at low values of the solid concentration (C (v)) and an asymptotic pattern at high C (v) were observed at N = 0.25 s(-1). These results suggest that mass transfer phenomena were commanded by the slurry properties and a semi-empirical equation of the form Sh = f(Re, Sc) seems to corroborate this finding. PMID- 17082915 TI - Penicillin acylase catalysis in the presence of ionic liquids. AB - Several ionic liquids were used as reaction media for penicillin G acylase catalysis. In all the assayed ionic liquids, [bmim]PF6 proved good media for PGA catalyzed hydrolysis. A novel [bmim]PF6/water two-phase system is provided for 6 aminopenicillanic acid (APA) production, which will be more benefical than aquous batch systems used widely in industrial production of APA. PMID- 17082916 TI - Characterization of the Caenorhabditis elegans G protein-coupled serotonin receptors. AB - Serotonin (5-HT) regulates a wide range of behaviors in Caenorhabditis elegans, including egg laying, male mating, locomotion and pharyngeal pumping. So far, four serotonin receptors have been described in the nematode C. elegans, three of which are G protein-coupled receptors (GPCR), (SER-1, SER-4 and SER-7), and one is an ion channel (MOD-1). By searching the C. elegans genome for additional 5-HT GPCR genes, we identified five further genes which encode putative 5-HT receptors, based on sequence similarities to 5-HT receptors from other species. Using loss-of-function mutants and RNAi, we performed a systematic study of the role of the eight GPCR genes in serotonin-modulated behaviors of C. elegans (F59C12.2, Y22D7AR.13, K02F2.6, C09B7.1, M03F4.3, F16D3.7, T02E9.3, C24A8.1). We also examined their expression patterns. Finally, we tested whether the most likely candidate receptors were able to modulate adenylate cyclase activity in transfected cells in a 5-HT-dependent manner. This paper is the first comprehensive study of G protein-coupled serotonin receptors of C. elegans. It provides a direct comparison of the expression patterns and functional roles for 5-HT receptors in C. elegans. PMID- 17082917 TI - Candidate pheromone binding proteins of the silkmoth Bombyx mori. AB - Pheromone reception is thought to be mediated by pheromone binding proteins (PBPs) in the aqueous lymph of the antennal sensilla. Recent studies have shown that the only known PBP of Bombyx mori (BmorPBP1) appears to be specifically tuned to bombykol but not to bombykal, raising the question of whether additional subtypes may exist. We have identified two novel genes, which encode candidate PBPs (BmorPBP2, BmorPBP3). Comparison with PBPs from various moth species have revealed a high degree of sequence identity and the three BmorPBP-subtypes can be assigned to distinct groups within the moth PBP family. In situ hybridization revealed that BmorPBP2 and BmorPBP3 are expressed only in relatively few cells compared to the number of cells expressing BmorPBP1. Double-labeling experiments have shown that the two novel BmorPBPs are expressed in the same cells but are not co-expressed with BmorPBP1. Furthermore, unlike BmorPBP1, cells expressing the newly identified PBPs did not surround neurons containing the BmOR-1 receptor. The results indicate that BmorPBP2 and BmorPBP3 are located in sensilla types, which are different from the long sensilla trichodea. PMID- 17082919 TI - Phosphorylation-dependent metal binding by alpha-synuclein peptide fragments. AB - Alpha-synuclein (alpha-syn) is the major protein component of the insoluble fibrils that make up Lewy bodies, the hallmark lesions of Parkinson's disease. Its C-terminal region contains motifs of charged amino acids that potentially bind metal ions, as well as several identified phosphorylation sites. We have investigated the metal-binding properties of synthetic model peptides and phosphopeptides that correspond to residues 119-132 of the C-terminal, polyacidic stretch of human alpha-syn, with the sequence Ac-Asp-Pro-Asp-Asn-Glu-Ala-Tyr-Glu Met-Pro-Ser-Glu-Glu-Gly (alpha-syn119-132). The peptide pY125 replaces tyrosine with phosphotyrosine, whereas pS129 replaces serine with phosphoserine. By using Tb(3+) as a luminescent probe of metal binding, we find a marked selectivity of pY125 for Tb(3+) compared with pS129 and alpha-syn119-132, a result confirmed by isothermal titration calorimetry. Truncated or alanine-substituted peptides show that the phosphoester group on tyrosine provides a metal-binding anchor that is supplemented by carboxylic acid groups at positions 119, 121, and 126 to establish a multidentate ligand, while two glutamic acid residues at positions 130 and 131 contribute to binding additional Tb(3+) ions. The interaction of other metal ions was investigated by electrospray ionization mass spectrometry, which confirmed that pY125 is selective for trivalent metal ions over divalent metal ions, and revealed that Fe(3+) and Al(3+) induce peptide dimerization through metal ion cross-links. Circular dichroism showed that Fe(3+) can induce a partially folded structure for pY125, whereas no change was observed for pS129 or the unphosphorylated analog. The results of this study show that the type and location of a phosphorylated amino acid influence a peptide's metal-binding specificity and affinity as well as its overall conformation. PMID- 17082918 TI - Characterization of a cyanobacterial-like uptake [NiFe] hydrogenase: EPR and FTIR spectroscopic studies of the enzyme from Acidithiobacillus ferrooxidans. AB - Electron paramagnetic resonance (EPR) and Fourier transform IR studies on the soluble hydrogenase from Acidithiobacillus ferrooxidans are presented. In addition, detailed sequence analyses of the two subunits of the enzyme have been performed. They show that the enzyme belongs to a group of uptake [NiFe] hydrogenases typical for Cyanobacteria. The sequences have also a close relationship to those of the H(2)-sensor proteins, but clearly differ from those of standard [NiFe] hydrogenases. It is concluded that the structure of the catalytic centre is similar, but not identical, to that of known [NiFe] hydrogenases. The active site in the majority of oxidized enzyme molecules, 97% in cells and more than 50% in the purified enzyme, is EPR-silent. Upon contact with H(2) these sites remain EPR-silent and show only a limited IR response. Oxidized enzyme molecules with an EPR-detectable active site show a Ni(r)*-like EPR signal which is light-sensitive at cryogenic temperatures. This is a novelty in the field of [NiFe] hydrogenases. Reaction with H(2) converts these active sites to the well-known Ni(a)-C* state. Illumination below 160 K transforms this state into the Ni(a)-L* state. The reversal, in the dark at 200 K, proceeds via an intermediate Ni EPR signal only observed with the H(2)-sensor protein from Ralstonia eutropha. The EPR-silent active sites in as-isolated and H(2)-treated enzyme are also light-sensitive as observed by IR spectra at cryogenic temperatures. The possible origin of the light sensitivity is discussed. This study represents the first spectral characterization of an enzyme of the group of cyanobacterial uptake hydrogenases. PMID- 17082920 TI - DNA binding by an imidazole-sensing CooA variant is dependent on the heme redox state. AB - CooA is a transcription factor from Rhodospirillum rubrum that is regulated by the binding of the small molecule effector, CO, to a heme moiety in the protein. The heme in CooA is axially ligated by two endogenous donors in the Fe(III) and Fe(II) states of the protein, and CO binding to the Fe(II) state results in replacement of the distal ligand. Reduction of the heme in the absence of CO results in a ligand switch on the proximal side, in which a cysteine thiolate in the Fe(III) state is replaced by a histidine in the Fe(II) state. Recently, a variant, termed RW CooA, was designed to respond to a new effector; Fe(II) RW CooA shows high specificity and induced DNA-binding activity in the presence of imidazole. Spectroscopic characterization of the imidazole adducts of RW CooA revealed that, unlike CO, imidazole binds to both Fe(III) RW CooA and Fe(II) RW CooA. The spectral characteristics are consistent with normal function of the redox-mediated ligand switch; Fe(III)-imidazole RW CooA bears a thiolate ligand and Fe(II)-imidazole RW CooA bears a neutral donor ligand. Since the effector binds to both redox states, RW CooA was used to probe the role of the redox mediated ligand switch in the CooA activation mechanism. Functional studies of Fe(III)-imidazole and Fe(II)-imidazole ligated RW CooA demonstrate that only the Fe(II)-imidazole form is active for DNA binding. Thus, the ligand switch is essential for the activating conformational change and may prevent aberrant activation of CooA by other neutral diatomic molecules. PMID- 17082921 TI - Size does matter: 1.5 Fr. stone baskets almost double irrigation flow during flexible ureteroscopy compared to 1.9 Fr. stone baskets. AB - With a new generation of flexible ureterorenoscopes, a new area in stone management is emerging. Limitation of vision with these new instruments is often caused by insufficient irrigation flow, especially when using instruments like stone baskets, resulting from partial obstruction of the working and irrigation channel with these instruments. Empirically, new available smaller stone baskets seem to dramatically improve irrigation and therefore vision in clinical use. The goal of this study was to show objective differences in basket diameters and flow rates in an in vitro setting. Diameters and irrigation flows in flexible ureterorenoscopes depending on different sizes of stone baskets (Fr. 1.5-1.7-1.9 2.2-2.4-3.0) and different deflections were measured. The measured diameter of the baskets varied within the first 20 cm and the true measured size varied from the manufacturer's specified size to a different extent. The new generation of 1.5 and 1.7 Fr. baskets improved irrigation flow, even compared to the smallest commonly used baskets, up to 68%. Interestingly, deflection did not influence irrigation flow. This study confirmed the subjective impression of inadequate description of relevant basket diameters as well as that of a significant improvement of irrigation flow with the newest generation of stone baskets with smaller diameters. PMID- 17082922 TI - Laser Doppler assessment of the influence of division at the root of the inferior mesenteric artery on anastomotic blood flow in rectosigmoid cancer surgery. AB - AIMS: The aim of this study is to evaluate the influence of dividing the inferior mesenteric artery (IMA) and preserving the left colic artery (LCA) on rectosigmoid cancer surgery. PATIENTS AND METHODS: Colonic blood flow at the proximal site of the anastomosis was measured by laser Doppler flowmetry in 96 patients with cancer of the rectum and sigmoid colon while clamping IMA or LCA. Results were analyzed with patient characteristics and postoperative complications. RESULTS: Blood flow was significantly decreased by either IMA or LCA clamping, and its reduction rate was 38.5 +/- 1.8%, ranged from 0 to 82.8%, or 16.4 +/- 1.8%, ranged from 0 to 66.2%, respectively. For multivariate analyses, aging and male gender were predictive factors of high blood flow reduction by IMA clamping. The reduction rate was significantly correlated with aging in male patients, while no such correlation was observed in women. Aging correlation in men was more significant in ultralow anterior resection cases. Three elderly male patients received IMA high ligation among 19 patients who demonstrated more than 50% blood flow reduction by IMA clamping. Among these, two patients, those who underwent ultralow anterior resection, suffered severe anastomotic ischemia. CONCLUSIONS: Colonic blood flow at the proximal site of the anastomosis was significantly decreased by either IMA or LCA clamping. Patients with high reduction by IMA clamping need intraoperative efforts to prevent anastomotic ischemia, particularly in elderly male patients who undergo ultralow anterior resection. PMID- 17082923 TI - Immunophenotyping and spatio-temporal distribution of aortic cell clusters in the bovine embryo. AB - In the present study the temporal and spatial appearance of aortic cell clusters in bovine embryos is described. Aorta-associated c-kit-positive cell clusters can be observed first in 23 days post inseminationem (dpi) bovine embryos and disappear after 34 dpi. For the first time, it was shown that the immunophenotype of these aortic cluster cells changes during embryonic development. Aortic cell clusters are c-kit+/CD45-/STA-, when they are first detected in the 23 dpi embryo, and acquire a c-kit+/CD45+/STA- phenotype in 27-29 embryos and a c kit+/CD45+/STA+ immunophenotype in 32-34-day-old specimens. Cell clusters are most prominent in the vicinity of lateral and ventral aortic branches, but rare in omphalomesenteric arteries and absent in Aa. umbilicales. Free c-kit-positive cells in an intravasal position are common, suggesting separation from the clusters in order to colonize subsequent hematopoietic organs, i.e., the liver and the mesonephros. Transmission electron microscopic analysis reveals the existence of primitive desmosomes between the clusters cells and adjacent endothelial cells as well as a fine basal lamina as a demarcation between the cluster cells and underlying mesenchymal cells. Material resembling extracellular matrix is found in large vacuoles in cluster cells of 23 dpi embryos. Immunocytochemistry reveals an intense accumulation of heparan sulfate proteoglycan and collagen IV in the aortic wall at the sites where cell clusters are attached. These observations suggest that the hematopoietic cell clusters induce the formation of a specific microenvironment within the aortic wall. PMID- 17082924 TI - N-halamine biocidal coatings. AB - Novel N-halamine siloxane and epoxide coatings are described. The coatings can be rendered biocidal by exposure to dilute bleach. Once the bound chlorine is lost from the coatings, it can be regenerated by further exposure to dilute bleach. Synthetic schemes and biocidal efficacy data are presented. The stabilities of the bound chlorine on the surfaces are also addressed. Substrates employed include sand, textiles, and paint. Potential uses for the technology are discussed. PMID- 17082925 TI - Expression of a beta-glucosidase gene results in increased accumulation of salicylic acid in transgenic Nicotiana tabacum cv. Xanthi-nc NN genotype. AB - A beta-glucosidase gene (bglA) from Butyrivibrio fibrisolvens H17c was cloned into the binary vector pGA482 under the control of the 35S Cauliflower Mosaic Virus (CaMV) promoter. A second construct was generated for accumulation of the bglA gene product in the vacuole of transformed tobacco plants. Reverse transcription-polymerase chain reaction analysis demonstrated that the bglA gene was expressed in 71% of cytosol-targeted and 67% of vacuole-targeted transgenic tobacco T(1) plants. T(1) transgenic plants (pGLU100 and pGLU200) exhibited elevated levels of free salicylic acid (SA) with a concomitant significant decrease in the level of glucosylsalicylic acid (GSA) compared to the untransformed tobacco plants and tobacco plants transformed with the empty vector (pGA482). Following inoculation with Tobacco Mosaic Virus (TMV), lesion area was 51% smaller in pGLU100 plants and 60% smaller in pGLU200 plants compared to inoculated untransformed and negative control plants. PMID- 17082926 TI - [What kind of stone prevention for whom? Risk-adjusted metaphylaxis following urinary stone disease]. AB - Approximately 5% of the German population suffers from urinary stone disease, but only 25% of these urolithiasis patients are at risk of recurrent stone disease or a severe metabolic disorder. It is important that patients at high risk are picked up early, so that appropriate therapy and measures designed to prevent secondary stone disease can be implemented. Risk classification is easily achieved by combining stone analysis with a basic diagnostic program. Patients at low risk need no further diagnostic evaluation or treatment, so that it is enough to recommend general metaphylaxis in these cases. In contrast, patients at high risk require additional specific aftercare and should be evaluated with the aid of a comprehensive diagnostic program from the start to allow precise definition of the metabolic targets. PMID- 17082928 TI - A lectin with antifungal and mitogenic activities from red cluster pepper (Capsicum frutescens) seeds. AB - A monomeric mannose/glucose-binding lectin, with a molecular mass of 29.5 kDa and an N-terminal sequence GQRELKL showing resemblance to that of the lectin-like oxidized low-density lipoprotein receptor from the rabbit, has been isolated from the seeds of red cluster pepper Capsium frutescens L. var. fasciculatum. The protocol involved anion exchange chromatography on diethylamino ethanol-cellulose and Q-Sepharose and fast protein liquid chromatography on Mono Q. Its hemagglutinating activity toward rabbit erythrocytes was inhibited by D: -mannose and glucose, specifically. The activity was stable from 0 to 40 degrees C, reached a maximum at pH 7 and 8, and was potentiated by Ca2+ and Mn2+ ions. The lectin showed strong mitogenic activity toward spleen cells isolated from BALB/c mice. The mitogenic activity, which reached a peak at a lectin concentration of 0.27 microM, was inhibited specifically by D(+)-mannose. The lectin was capable of inhibiting the germination of Aspergillus flavus and Fusarium moniliforme spores and hyphal growth in the two fungi. PMID- 17082929 TI - Characterization, cloning, sequencing, and expression of an aminopeptidase N from Streptomyces sp. TH-4. AB - The aminopeptidase N (TH-4AP) of Streptomyces sp. TH-4 was purified from a culture supernatant. The purified enzyme had a molecular mass of 95 kDa. The gene encoding TH-4AP was cloned and sequenced. The primary structure of the protein possessed the PepN-conserved motif GxMEN and the zinc-binding motif HExxHx18E, and showed 88% identity with that of PepN from Streptomyces lividans strain 66. We succeeded in overproducing a His-tagged recombinant enzyme using Escherichia coli. The enzyme had a 1.5-fold higher activity in the presence of cobalt ions than in their absence. To evaluate the possible application of TH-4AP to decrease the content of bitter peptides, we investigated the ability of Streptomyces aminopeptidases to hydrolyze synthetic peptides by a coupling method using L amino acid oxidase and peroxidase. The substrate specificity of TH-4AP toward synthetic peptides was significantly different from that toward aminoacyl-p nitroanilide derivatives. PMID- 17082927 TI - [Pharmacotherapy for schizophrenia]. AB - The management of schizophrenia patients remains one of the great challenges in psychiatry. Despite the undisputed effectiveness of antipsychotic drugs, patients and their physicians still face considerable difficulties mainly related to incomplete or lacking treatment response and the inability to predict the individual efficacy and tolerability. In this manuscript we review the key elements of pharmacological treatment of this disorder, encompassing acute and long-term management as well as specific management problems ranging from acutely violent patients to treatment-resistant subjects. Along with general treatment principles, the document provides specific information regarding efficacy and safety features of antipsychotics. Many of the currently available treatment recommendations/guidelines are based on the evidence reviewed here. This review is meant to serve as a guide for clinicians involved in managing schizophrenia, whether in a psychiatric hospital setting or as family physicians in private practice. PMID- 17082930 TI - Production of 7, 10-dihydroxy-8(E)-octadecenoic acid from triolein via lipase induction by Pseudomonas aeruginosa PR3. AB - Hydroxy fatty acids (HFA) have gained importance because of their special properties such as higher viscosity and reactivity compared with other non hydroxy fatty acids. The bacterial isolate Pseudomonas aeruginosa (PR3) was reported to produce mono-, di-, and trihydroxy fatty acids from different unsaturated fatty acids. Of those, 7,10-dihydroxy-8(E)-octadecenoic acid (DOD) was produced with high yield from oleic acid by PR3. Up to now, the substrates used for microbial HFA production were free fatty acids. However, it is possible to utilize triacylglycerides, specifically triolein containing three oleic groups, as a substrate by microbial enzyme system involved in HFA production from oleic acid. In this study we used triolein as a substrate and firstly report that triolein could be efficiently utilized by PR3 to produce DOD. Triolein was first hydrolyzed into oleic acid by the triolein-induced lipase and then the released oleic acid was converted to DOD by PR3. Results from this study demonstrated that natural vegetable oils, without being intentionally hydrolyzed, could be used as efficient substrates for the microbial production of value-added hydroxy fatty acids. PMID- 17082931 TI - Ecological and biotechnological aspects of lichens. AB - Lichens and the partners from three different kingdoms are both taxonomically and physiologically a very diverse group, which makes them interesting from both ecological and biotechnological points of view. A lichen is a mutual ecophysiological innovation in many extreme environments in which symbiosis seems to protect the partners. Lichen's ability to grow in harsh environments can be advantageous, resulting in important ecological niches, or disadvantageous when lichens occupy and cause biodeterioration of cultural monuments. Recently, new candidate compounds for drugs, UVB protection, and antifreeze proteins for frozen foods were discovered. Lichens were also found to have potential in bioplastic degradation and prevention of desertification. Nevertheless, there is still large potential for further industrial screening and research on lichen products. Due to improved culture techniques of isolated symbionts, increased knowledge of their secondary metabolism and improved methods for solubilizing lichen metabolites, the screening and activity tests can be implemented more easily today than in the past. PMID- 17082932 TI - Trichuris trichiura. PMID- 17082933 TI - Markedly hypoplastic circumflex retroesophageal right aortic arch: MR imaging and surgical implications. AB - BACKGROUND: Circumflex retroesophageal right aortic arch with a markedly hypoplastic retroesophageal segment is a rare anomaly of the aortic arch. Adequate surgical management relies on precise diagnosis, which might not be feasible with echocardiography. OBJECTIVE: To demonstrate the utility of MR imaging in establishing the diagnosis and to describe potential pitfalls in making the diagnosis. MATERIALS AND METHODS: Three patients with a circumflex retroesophageal right aortic arch with a markedly hypoplastic retroesophageal segment were retrospectively evaluated. All patients underwent evaluation by echocardiography and MR imaging. The MR imaging consisted of 3-D MR angiography in two patients and fast gradient recalled echo with cardiac-triggered segmented acquisition in one patient. Surgical confirmation was obtained on all three patients. RESULTS: The arch anatomy was accurately depicted in all three patients by MR imaging and in none of the patients by echocardiography. CONCLUSION: MR imaging is extremely useful in establishing the diagnosis of markedly hypoplastic retroesophageal circumflex right aortic arch and thus helps in surgical planning. PMID- 17082934 TI - Congenital pyriform aperture stenosis. AB - Nasal airway obstruction is a potentially life-threatening condition in the newborn. Neonates are obligatory nasal breathers. The pyriform aperture is the narrowest, most anterior bony portion of the nasal airway, and a decrease in its cross-sectional area will significantly increase nasal airway resistance. Congenital nasal pyriform aperture stenosis (CNPAS) is a rare, unusual form of nasal obstruction. It should be considered in the differential diagnosis of any neonate or infant with signs and symptoms of upper airway compromise. It is important to differentiate this level of obstruction from the more common posterior choanal stenosis or atresia. CNPAS presents with symptoms of nasal airway obstruction, which are often characterized by episodic apnea and cyclical cyanosis. PMID- 17082935 TI - Significance of hypogonadism in erectile dysfunction. AB - To review the role and significance of hypogonadism, defined as a low testosterone (T) level, in erectile dysfunction (ED). Review of literature. Serum T is below 3 ng/ml in 12% of ED patients, including 4% before and 15% after the age of 50. Replacement studies in men with severe hypogonadism demonstrate that sexual desire and arousal, as well as the frequency of sexual activity and spontaneous erections are clearly T-dependant. Psychic erections are partly T dependant. The effects of T upon sexual function are dose-dependant up to a threshold level that is consistent within an individual, but markedly variable between individuals, ranging from 2 to 4.5 ng/ml. More evidence is required to confirm a significant impact of T on the intrapenile vascular mechanisms of erections in men as it is the case in animals. No convincing association of T with ED has been found in epidemiological studies. As concerns clinical experience, although a meta-analysis of the randomized controlled trials established that T therapy consistently restores erectile function in young hypogonadal patients with T below 3.46 ng/ml, the effects of this treatment have been mostly disappointing when used alone in older patients consulting for ED who are subsequently diagnosed to have hypogonadism following routine T measurement. These poor results may probably be explained by the high prevalence of co morbidities, and by the fact that ED itself may induce hypogonadism. Combination therapy with T and PDE5 inhibitor (PDE5I) may be effective in the hypogonadal ED patients when T therapy alone fails. However, more evidence is required to confirm the hypothesis that a minimum level of T is required for a complete effect of PDE5I in certain men, since a PDE5I was able to restore complete erections in severely hypogonadal men. Though a low T level is not always the only cause of ED in hypogonadal ED patients, there are important benefits in screening for hypogonadism in ED. A low T level justifies a 3 month trial of T therapy, before combining a PDE5I if T therapy alone fails. PMID- 17082936 TI - The male biological clock. AB - Do men have biological clocks that affect their hormone levels, fertility, and the genetic quality of their sperm? Women can no longer be viewed as solely responsible for age-related fertility and genetic problems. The effects of andropause and advanced paternal age on fertility and offspring are still under investigation. Further research is needed to fully characterize the associated risks and to treat the underlying abnormalities. A better understanding of the cellular and biochemical mechanisms of "gonadal" aging is important in order to determine safe, effective ways to delay this process and "rewind" the male biological clock. The benefits may include decreasing the potential for adverse genetic consequences in offspring, improvement in the sexual and reproductive health of aging males, and increase a woman's chance of having healthy children by correcting defects in the male reproductive system. PMID- 17082937 TI - Understanding the needs and objectives of erectile dysfunction patients. AB - Consideration of the needs and objectives of individual patients significantly influences the behavior of erectile dysfunction (ED) patients and their partners. Evaluation of an erectile dysfunction (ED) patient requires individualized consideration of the expectations, needs, and objectives of the affected couple. These factors can influence attitudes toward diagnosis, treatment seeking, and ultimately treatment compliance and dropout. Relevant ED market research studies were presented at the 2006 annual meeting of the American Urological Association. The studies explored the prevalence of ED, access to treatment, and ED patient attitudes and preferences. Several barriers continue to influence treatment seeking behavior in men with ED. These barriers have had a negative impact on the ED patient and marketplace resulting in low rates of utilization and high rates of dropout from therapies for ED. PMID- 17082938 TI - Retarded ejaculation. AB - Retarded ejaculation (RE) has a relatively low prevalence (<3%), yet this condition results in considerable distress, anxiety, and lack of sexual confidence for those suffering from it. Furthermore, men with partners often experience impairment of both the sexual and nonsexual aspects of their relationships, with such negative effects compounded when procreation is a consideration. The definition of RE is ambiguous, due to the variability and paucity of data regarding normal coital ejaculatory latency. RE is influenced by both biogenic and psychogenic components, which may vary over time both between and within individuals. While specific pathophysiology can often be identified, further elucidation of the biogenic components of this dysfunction will require greater understanding of the physiological mechanisms underlying ejaculation. Yet, the most useful strategies for understanding RE will integrate rather than isolate the various biogenic and psychogenic aspects of this dysfunction. Evidence based evaluation and treatment protocols for this disorder are lower than for other sexual dysfunctions, but reports suggest better treatment efficacy when the etiology is predominantly psychogenic. As with erectile dysfunction (ED) and premature ejaculation (PE), if safe and efficacious oral pharmaceuticals are eventually developed for this condition, the treatment algorithm is likely to undergo significant alteration. Even then, however, the most effective treatments are likely to result from a combination treatment that integrates sex coaching with pharmacotherapy. PMID- 17082939 TI - Female sexual dysfunction, voiding symptoms and depression: common findings in partners of men with erectile dysfunction. AB - The aim of this study was to investigate the prevalence of female sexual dysfunction (FSD), urinary symptoms, and depressive symptoms in female partners of men presenting with erectile dysfunction (ED). A multi-component questionnaire was administered to female partners of men with erectile dysfunction presenting to a urology center. It contained a standardized sexual function component (the Brief Index of Sexual Function for Women), a depression scale (Centers for Epidemiologic Studies-Depression, CES-D), a demographics questionnaire and a general medical questionnaire. A total of 73 consecutive female partners of male patients presenting with ED, were surveyed using the questionnaire at their counterpart's visit. Fifty-two women responded, of whom 50 filled out the questionnaire adequately for proper evaluation. This indicated a response rate of 68% (50/73). The mean age was 44.8 years (range 20.0-83.0). Thirty-eight of the 50 women (76%) reported being sexually active. A variety of sexual behaviors were reported including 40% (20/50) of women engaging in vaginal intercourse. Sexual dysfunction symptoms included: anxiety/inhibition (26%), hypoactive desire (20%), arousal/lubrication difficulty (30%), orgasmic difficulty (24%), dyspareunia (18%), incontinence during intercourse (8%), and sexual dissatisfaction (34%). Eight women (16%) reported difficulty communicating sexual issues with their partners. Forty-one women (82%) rated sexual activity as an important part of their lives. Urinary symptoms of frequency and urgency were reported by 18/50 (36%). Depressive symptoms were present in 22/50 (44%). FSD disorders, urinary symptoms and depressive symptoms are common in partners of men with erectile dysfunction. PMID- 17082940 TI - Bladder-sparing approaches to invasive disease. AB - Although immediate radical cystectomy remains the standard of care for invasive bladder cancer, a large body of international experience from single institutions and cooperative groups has accumulated, suggesting favorable results with bladder sparing approaches in appropriately selected patients. Modern selective bladder preservation with trimodality therapy, consisting of transurethral resection of the bladder tumor, radiation, and chemotherapy, can achieve complete response rates of 60-80%, 5-year survival rates of 50-60%, and survival rates with an intact bladder of 40-45%. Although no randomized comparisons between cystectomy and trimodality therapy exist, long-term data confirm that the 10-year overall and disease-specific survival rates for patients in bladder-sparing protocols are comparable to outcomes reported in contemporary cystectomy series. In addition, quality of life studies have demonstrated that the retained native bladder functions well. Thus, trimodality therapy with careful cystoscopic surveillance and with prompt cystectomy for invasive recurrences has emerged as a legitimate alternative to extirpative surgery. Future work will continue to optimize the bladder-sparing regimen while limiting toxicity. PMID- 17082941 TI - Sclerotherapy of cervical cysts with Picibanil (OK-432). AB - The effectiveness of intralesional sclerotherapy of lymphangiomas and ranulas with OK-432 (Picibanil) has been proved in several clinical studies. The aim of our study was to review the effectiveness of sclerotherapy of benign cervical cysts with Picibanil as an alternative method to surgical excision. Between March 2002 and March 2006, a prospective observational study was carried out to assess the effects of Picibanil on cervical cysts. Between 2002 and 2006 we treated 14 patients having cervical cysts through intralesional application of Picibanil with a dose of 0.01 mg/ml. So far we used Picibanil with 13 patients achieving a high success rate. In eight cases we observed, both clinically and ultrasonographically, a nearly complete regression, and a complete regression of the cysts in three cases. In two cases the cysts atrophied. In these cases only residual findings could be observed. In one case we extirpated the remaining cyst. If there is no clear reaction of the cyst to the treatment, an excision is indicated 6 weeks after the injections to gain meaningful histological examination. No significant complication after sclerotherapy with Picibanil was observed. According to our results the application of OK-432 (Picibanil) is a safe and effective primary method for sclerotherapy of benign cervical cysts which can replace surgical extirpation in special cases. However, the risk of malign diseases has to be excluded before the commencement of the Picibanil treatment. PMID- 17082942 TI - Presentation of two cases of nasal type NK/T-cell lymphoma. AB - Nasal type NK/T-cell lymphoma is a rare type of predominantly extranodal non Hodgkin lymphoma. Early and correct diagnosis with prompt treatment of NK/T-cell lymphoma is important in view of its potentially aggressive behavior and poor response to treatment with additional sites of tumor developing sometimes weeks or months after initial diagnosis. Unfortunately diagnosis of NK/T-cell lymphomas often proves difficult. The diagnosis is essentially based on the clinical presentation of extranodal ulcerative lesions in the upper aero-digestive tract and histopathologic analysis of biopsies using immunohistochemistry. Here we present two cases with nasal-type NK/T-cell lymphoma that illustrate that definitive diagnosis is often delayed due to the atypical initial presentation. We will discuss the pitfalls in diagnosing this rare type of neoplasm and review the treatment options. PMID- 17082943 TI - The cochleogram of the guinea pig. AB - The cochleogram is an important tool to relate properties of the cochlea (e.g. hair cell loss, damaged hair cells) to their position in the cochlear turns, to calculate the average hair cell density, and to measure the length of the whole cochlea. In this work different methods of plotting cochleograms are compared. We suggest that a sector-wise division of the cochlea for counting a cochleogram has advantages over line diagrams that provide a higher spatial resolution but might lead to misinterpretations of the degree of missing hair cells. The scanning electron microscopic analysis of 171 guinea pig cochleas revealed a mean basilar membrane length of 16.4 +/- 1.4 mm (mean +/- standard deviation) with sector lengths of 6.9, 4.2, 3.2, and 1.9 mm, thus adding relevant information to the morphology of the guinea pig cochlea. PMID- 17082944 TI - Cochlear implantations in visually impaired patients. AB - We retrospectively review the cases to evaluate the outcome of cochlear implantation (CI) in patients with severe-to-profound hearing loss and visual impairment (VI). Six adults with severe or profound hearing loss and significant VI underwent multichannel CI. Follow-up period ranged from 17 months to 7 years. Case history, etiology of visual and hearing loss, and benefit from CI were evaluated. To measure the outcomes, we selected the pure-tone thresholds with CI, the speech discrimination scores (SDS) using the Japanese video SDS system, the speech perception rates using the Japanese CD SDS system by monosyllable and word, and the open-set and closed sentence score using live voice. All the patients live happily after CI. There was no significant difference between the present six patients and the patients with profound hearing loss without VI in evaluations of hearing and quality of life. CI can play a significant rehabilitative role in patients with severe hearing loss and VI. PMID- 17082945 TI - Petrous bone cholesteatoma: clinical longitudinal study. AB - The object of this retrospective study was to describe a series of patients with petrous bone cholesteatomas, paying particular attention to classification, diagnosis, surgical strategy, results, complications and recurrences. Furthermore, the study was designed to evaluate the impact of imaging techniques on an early diagnosis. Topographically, the petrous bone cholesteatomas of the present series were grouped using Sanna's classification and different surgical approaches were used. High resolution CT and/or MRI were used to follow-up the patients. The case notes of 52 patients with petrous bone cholesteatomas who were referred to our hospital for surgery between 1987 and 2003 were reviewed postoperatively. There were 45 primary cases and 7 recurrences. The facial nerve had been infiltrated and compressed by the cholesteatoma in 18 patients. Fourteen were managed with cable grafts using sural nerve or great auricular nerves. About 26 patients with preoperative grade I confirmed their normal facial function in 23 cases. In the other ten patients, the preoperative facial paralysis was due to compression by the cholesteatoma and its removal provided partial recovery of facial function in four patients. Our study compared two observation periods (1987-1996 and 1997-2003) when the diffusion and the availability of imaging techniques in our national health system had considerably increased. Two important factors emerged: firstly, the number of less extensive surgical approaches was higher in the more recent observation period, proving that cholesteatomas smaller in size had been diagnosed. Secondly, preoperative facial paralysis was less frequent in the same period-falling to 25% of cases of total facial paralysis from the 45.8% of the earlier period-practically half as much. The partial paralyses instead increased slightly, demonstrating that otologists have become more sensitive to and pay more attention to this symptom. PMID- 17082946 TI - Short-term results using Kurz titanium ossicular implants. AB - The efficiency of titanium middle ear prosthesis for ossicular reconstruction in chronic ear disease is investigated in a Scandinavian two-center retrospective study from a Norwegian tertiary otology referral center and a Finnish otology referral center. Retrospective chart reviews were performed for procedures involving 73 titanium prostheses between 1999 and 2004. All patients that underwent surgery including the Kurz Vario titanium prosthesis were included in the study, 38 procedures including the partial ossicular replacement prosthesis (PORP) and 35 procedures including the total ossicular replacement prosthesis (TORP). Mean follow-up was 14 months. The ossiculoplasty was performed alone (29 patients) or in combination with other chronic ear surgery procedures (34 patients). Comparisons of preoperative and postoperative pure tone averages (0.5, 1, 2, and 3 kHz) according to AAO-HNS guidelines are presented, as well as data for different PTA definitions. Otosurgery procedures, complications, revisions, and extrusion rates are reported. A postoperative air-bone gap (ABG) of or = 4.0. At pH< or =3.5, concentrations of the AP increased from the beginning of the experiments to day 5, then decreased from day 5 to 15, and finally increased from day 15 to the end of the experiments. At pH > or = 4.0, concentrations of the AP increased consecutively from the beginning of the experiments to day 10 and decreased from day 10 to the end of the experiments. Such a finding is useful for agricultural practices since soil P is one of the most important macronutrients for plant growth. In general, SOM content decreased with time as the Latosol was leached by the SAR at all pH levels. A maximum concentration of soil fulvic acid was found after 15 days of the experiments due to the degradation of the SOM. A multiple regression analysis showed that a very strong relationship was obtained between the soil AP and the other three parameters (i.e., pH, SOM, and sorption P). PMID- 17083001 TI - Evaluation of developmental toxicity of amitraz in Sprague-Dawley rats. AB - This study investigated the potential adverse effects of amitraz on the initiation and maintenance of pregnancy in Sprague-Dawley rats as well as its effects on embryo-fetal development after maternal exposure during the entire pregnancy period. Amitraz was administered to pregnant rats by gavage from days 1 to 19 of gestation at dose levels of 0, 3, 10, and 30 mg/kg/day. All dams underwent a caesarean section on day 20 of gestation and their fetuses were examined for any external, visceral, and skeletal abnormalities. At 30 mg/kg, maternal toxicity manifested as an increase in the incidence of abnormal clinical signs and a lower body weight gain and food intake. Developmental toxicity included an increase in the fetal death rate, a decrease in the litter size, and a reduction in the fetal body weight. In addition, there was an increase in the incidence of fetal external, visceral, and skeletal abnormalities. At 10 mg/kg, maternal toxicity observed included a decrease in the body weight gain and a decrease in food intake. In addition, minimal developmental toxicity, including a decrease in the fetal body weight, an increase in the visceral and skeletal aberrations, and a delay in fetal ossification. There were no signs of either maternal toxicity or developmental toxicity at 3 mg/kg. These results show that amitraz administered during the entire pregnancy period in rats is embryotoxic and teratogenic at the maternally toxic dose (i.e., 30 mg/kg/day) and is minimally embryotoxic at a minimally maternally toxic dose (i.e., 10 mg/kg/day). Under these experimental conditions, the no-observed-adverse-effect level of amitraz for both dams and embryo-fetal development is estimated to be 3 mg/kg/day. PMID- 17083002 TI - Mercury exposure and effects on cavity-nesting birds from the Carson River, Nevada. AB - Mercury (Hg) concentrations were 15-40 times higher in the eggs and livers of tree swallows (Tachycineta bicolor) and house wrens (Troglodytes aedon) that nested along the Carson River at and below Dayton, Nevada than in the same species above the mining-impacted areas. Hg contamination was mainly the result of processing mills in the 1800s that used Hg to separate gold and silver from ore. The exposure pattern of tree swallows and house wrens along the Carson River was consistent with their trophic status (i.e., lower levels in liver tissue of aquatic insectivores than in piscivorous birds nesting nearby). Even though they are aquatic insectivores, tree swallows and house wrens were exposed to the same amount of Hg as piscivores in the Florida Everglades; this indicated the extreme level of Hg contamination in the Carson River. Only 70-74% of the eggs hatched. This was less than the nationwide average for these two species that generally hatch > or =85% of eggs. Although the sample size was small, Hg might be impacting reproductive end points in cavity-nesting birds from the Carson River. Other trace elements were present at background concentrations. PMID- 17083003 TI - A mathematical modelling for the cheliped regeneration with handedness in fiddler crab. AB - An enormously developed giant cheliped with the other small one characterizes the adult male fiddler crab. Some experiments with artificial severances of cheliped indicate that such a handedness in the cheliped size is maintained even after the regeneration of severed cheliped. Other experimental researches give some results about an unknown physiological system which controls the emergence and the regeneration of the handedness in the cheliped size. In this paper, with two hypothesized factors relevant to the regeneration of a severed cheliped, we propose a simple mathematical model to describe the experimental result about the cheliped regeneration with a handedness after the cheliped severance for the fiddler crab. Our model gives a suggestion about an underlying system for the cheliped regeneration in the fiddler crab or some other crustacean species. PMID- 17083004 TI - Information theory in living systems, methods, applications, and challenges. AB - Living systems are distinguished in nature by their ability to maintain stable, ordered states far from equilibrium. This is despite constant buffeting by thermodynamic forces that, if unopposed, will inevitably increase disorder. Cells maintain a steep transmembrane entropy gradient by continuous application of information that permits cellular components to carry out highly specific tasks that import energy and export entropy. Thus, the study of information storage, flow and utilization is critical for understanding first principles that govern the dynamics of life. Initial biological applications of information theory (IT) used Shannon's methods to measure the information content in strings of monomers such as genes, RNA, and proteins. Recent work has used bioinformatic and dynamical systems to provide remarkable insights into the topology and dynamics of intracellular information networks. Novel applications of Fisher-, Shannon-, and Kullback-Leibler informations are promoting increased understanding of the mechanisms by which genetic information is converted to work and order. Insights into evolution may be gained by analysis of the the fitness contributions from specific segments of genetic information as well as the optimization process in which the fitness are constrained by the substrate cost for its storage and utilization. Recent IT applications have recognized the possible role of nontraditional information storage structures including lipids and ion gradients as well as information transmission by molecular flux across cell membranes. Many fascinating challenges remain, including defining the intercellular information dynamics of multicellular organisms and the role of disordered information storage and flow in disease. PMID- 17083005 TI - Modeling tick-borne disease: a metapopulation model. AB - Recent increases in reported outbreaks of tick-borne diseases have led to increased interest in understanding and controlling epidemics involving these transmission vectors. Mathematical disease models typically assume constant population size and spatial homogeneity. For tick-borne diseases, these assumptions are not always valid. The disease model presented here incorporates non-constant population sizes and spatial heterogeneity utilizing a system of differential equations that may be applied to a variety of spatial patches. We present analytical results for the one patch version and find parameter restrictions under which the populations and infected densities reach equilibrium. We then numerically explore disease dynamics when parameters are allowed to vary spatially and temporally and consider the effectiveness of various tick-control strategies. PMID- 17083006 TI - Prognosis scores of Tokuhashi and Tomita for patients with spinal metastases of renal cancer. AB - BACKGROUND: Retrospective evaluation of the prognosis scores of Tokuhashi and Tomita for life expectancy in 37 consecutive patients with spinal metastases secondary to renal cancer who underwent surgery. The score of Tokuhashi, composed of six parameters, each rated from zero to two, has been proposed in 1990 for the prognostic assessment of patients with spinal metastases. In 2001, Tomita et al. created another prognostic score, composed of three parameters, growth behaviour of the primary tumor (slow, moderate and rapid) and the evidence of visceral and bony metastases. METHODS: Thirty-seven patients, surgically treated for vertebral metastases secondary to renal cancer were studied. The scores according to Tokuhashi and Tomita were calculated for each patient. RESULTS: Applying the Tokuhashi Score for the estimation of life expectancy of renal cancer patients with vertebral metastases was found to provide very reliable results with a statistically high significance. The analysis according to Tomita showed no correlation between predicted and real survival. The statistical analysis did not show any significance. CONCLUSION: For surgical decisions in renal cancer patients with spinal metastases, the prognostic score of Tokuhashi appears to be much more valuable than the Tomita score. PMID- 17083007 TI - Sympathectomy for hyperhidrosis: should we place the clamps at T2-T3 or T3-T4? AB - Endoscopic thoracic sympathectomy is routinely used to treat severe hyperhidrosis. It is usually performed at the T2-T3 level of the nerve, but may produce less severe compensatory hidrosis if performed at a lower level. This study evaluates the outcome of 1,274 patients who underwent endoscopic thoracic sympathectomy for plamar, plantar, axillary or facial hyperhidrosis/blushing. Half of the patients were clamped at the T2-T3 level and half were clamped at the T3-T4 level. Postsurgical symptoms and side effects were assessed by interview. All of patients with palmar hyperhidrosis were cured or improved. Patients with plantar and axillary hyperhidrosis were more likely to be improved at T3-T4 level clamping. Patients with facial hyperhidrosis were more likely to be cured at T2 T3 level, but did show improvement at the T3-T4 level. Overall satisfaction was higher in the T3-T4 group. Some degree of mild compensatory sweating occurred in all patients. However, severe compensatory sweating was more common in the T2-T3 group. Around 2% of patients requested a reversal of their surgery. Endoscopic thoracic sympathectomy is a safe and effective treatment for hyperhidrosis. Clamping at the T3-T4 level has a more successful outcome. In particular, it appears to reduce the incidence of severe compensatory hidrosis. PMID- 17083008 TI - Subjective and objective risk as predictors of influenza vaccination during the vaccine shortage of 2004-2005. AB - BACKGROUND: We aimed to identify the role of objective risk status and subjective risk beliefs in influenza vaccination decisions during the recent rationing of influenza vaccine. METHOD: A random sample of 300 Americans, obtained through random-digit dialing, was interviewed regarding influenza vaccination practices and beliefs in September 2004 and again in March 2005. RESULTS: One-half of individuals at high risk of influenza did not know that they were at high risk and, therefore, were not vaccinated. Respondents at high objective risk were more likely to report having been vaccinated than respondents who were not at high objective risk (36% vs. 6%, respectively; odds ratio, 8.31; 95% confidence interval, 3.65-18.88). However, a more powerful predictor of self-reported vaccination was subjective risk (64% vs. 7%, respectively; odds ratio, 24.02; 95% confidence interval, 12.18-48.09). Subjective risk fully mediated the relationship between objective risk and vaccination. Other predictors of vaccination included physician recommendation, habit, prior vaccination intention, belief that the influenza vaccine is safe and effective, perceived likelihood of getting influenza, and trait neuroticism. CONCLUSION: Health communication efforts must be more effective in persuading adults with chronic illness and individuals in contact with persons at risk that they should be vaccinated against influenza. PMID- 17083009 TI - Serogroup C meningococcal glycoconjugate vaccine in adolescents: persistence of bactericidal antibodies and kinetics of the immune response to a booster vaccine more than 3 years after immunization. AB - BACKGROUND: The persistence of protection from meningococcal disease following immunization with serogroup C meningococcal (MenC) glycoconjugate vaccines in infancy is short-lived. The duration of protective immunity afforded by these vaccines in other at-risk age groups (i.e., adolescents and young adults) is not known. We evaluated the persistence of bactericidal antibodies following immunization with a MenC glycoconjugate vaccine (MenCV) in adolescents and the kinetics of immune response to a meningococcal AC plain polysaccharide vaccine (MenPS) challenge or a repeat dose of MenCV. METHODS: We conducted a randomized comparative trial of 274 healthy 13-15-year-olds from whom a total of 4 blood samples were obtained (prior to administration of a dose of MenPS or MenCV, again on 2 further occasions at varying times from days 2-7 after vaccination, and finally on day 28 after vaccination. The correlate of protection was a serum bactericidal assay titer > or = 8 (with a serum bactericidal assay using human complement). RESULTS: A serum bactericidal assay using human complement titer > or = 8 was observed in 75% of participants at baseline (mean age, 14.5 years; mean time since routine MenCV vaccination, 3.7 years). No increase in serum bactericidal assay geometric mean titers was detected until day 5 after administration of MenPS. Geometric mean titers following administration of MenCV were significantly higher than those observed following administration of MenPS, at days 5, 7, and 28. CONCLUSIONS: This study showed sustained levels of bactericidal antibodies for at least 3 years after immunization of adolescents with MenCV. After challenge of immunized adolescents with MenPS, there was no increase in serum bactericidal assay observed until day 5 after vaccination, indicating that immunological memory may be too slow to generate protection against this potentially rapidly invasive organism. PMID- 17083010 TI - Vaccine prevention of meningococcal disease: making slow progress. PMID- 17083011 TI - Reduced ability of penicillin to eradicate ingested group A streptococci from epithelial cells: clinical and pathogenetic implications. AB - BACKGROUND: Group A streptococci (Streptococcus pyogenes; GAS) invades human epithelial cell lines. Failure of penicillin to eradicate GAS from the throats of patients, especially those who are GAS "carriers," has been increasingly reported. However, there has been no comprehensive evaluation of how effectively antibiotics that are used to treat GAS enter upper respiratory tract epithelial cells and kill internalized GAS. We examined the viability of ingested, intracellular GAS after epithelial cell exposure to antibiotics commonly recommended for therapy of GAS infections. METHODS: A human laryngeal epithelial cell line (HEp-2) was used. Three techniques were used to study antibiotic (penicillin V, erythromycin, azithromycin, cephalothin, and clindamycin) killing of ingested GAS: examination by electron microscopy of ultrathin sections of ingested GAS, qualitative determination of intra-epithelial cell antibiotic, and special stain evaluation of intracellular GAS viability after epithelial cell exposure to antibiotics. RESULTS: GAS survived intracellularly despite exposure of the GAS-containing epithelial cells to penicillin. In contrast, there was killing of ingested GAS after exposure of epithelial cells to either erythromycin or azithromycin. Electron microscopy confirmed a lack of intracellular GAS fragmentation (cell death) after exposure of epithelial cells to penicillin in contrast to obvious GAS fragmentation after epithelial cell exposure to erythromycin or azithromycin. Cephalothin, a cephalosporin, and clindamycin were more effective in killing ingested GAS than was penicillin, but they were less effective than erythromycin or azithromycin. CONCLUSIONS: These observations strongly suggest that if the GAS upper respiratory tract carrier state results from intra-epithelial cell GAS survival, the failure of penicillin to kill ingested GAS may be related to a lack of effective penicillin entry into epithelial cells. These unique observations may have clinical implications for understanding GAS respiratory tract carriers and managing GAS infections. PMID- 17083012 TI - Bacteremia due to extended-spectrum beta -lactamase-producing Escherichia coli in the CTX-M era: a new clinical challenge. AB - BACKGROUND: Extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli, particularly those producing CTX-M types of ESBL, are emerging pathogens. Bacteremia caused by these organisms represents a clinical challenge, because the organisms are frequently resistant to the antimicrobials recommended for treatment of patients with suspected E. coli sepsis. METHODS: A cohort study was performed that included all episodes of bloodstream infection due to ESBL producing E. coli during the period from January 2001 through March 2005. Data on predisposing factors, clinical presentation, and outcome were collected. ESBLs were characterized using isoelectric focusing, polymerase chain reaction, and sequencing. RESULTS: Forty-three episodes (8.8% of cases of bacteremia due to E. coli) were included; 70% of the isolates produced a CTX-M type of ESBL. The most frequent origins of infection were the urinary (46%) and biliary tracts (21%). Acquisition was nosocomial in 21 cases (49%), health care associated in 14 cases (32%), and strictly community acquired in 8 cases (19%). Thirty-eight percent and 25% of patients had obstructive diseases of the urinary and biliary tracts, respectively, and 38% had recently received antimicrobials. Nine patients (21%) died. Compared with beta-lactam/beta-lactamase-inhibitor and carbapenem-based regimens, empirical therapy with cephalosporins or fluoroquinolones was associated with a higher mortality rate (9% vs. 35%; P=.05) and needed to be changed more frequently (24% vs. 78%; P=.001). CONCLUSIONS: ESBL-producing E. coli is a significant cause of bloodstream infection in hospitalized and nonhospitalized patients in the context of the emergence of CTX-M enzymes. Empirical treatment of sepsis potentially caused by E. coli may need to be reconsidered in areas where such ESBL-producing isolates are present. PMID- 17083013 TI - Extended-spectrum beta-lactamases in multidrug-resistant Escherichia coli: changing the therapy for hospital-acquired and community-acquired infections. PMID- 17083014 TI - Microbiology of acute otitis media in children with tympanostomy tubes: prevalences of bacteria and viruses. AB - BACKGROUND: Bacteria are found in 50%-90% of cases of acute otitis media (AOM) with or without otorrhea, and viruses are found in 20%-49% of cases. However, for at least 15% of patients with AOM, the microbiological etiology is never determined. Our aim was to specify the full etiology of acute middle ear infection by using modern microbiological methods concomitantly for bacterial and viral detection. METHODS: The subjects were 79 young children having AOM with new onset (<48 h) of otorrhea through a tympanostomy tube. Middle ear fluid samples were suctioned from the middle ear through the tympanostomy tube. Bacteria were sought by culture and polymerase chain reaction; viruses were analyzed by culture, antigen detection, and polymerase chain reaction. RESULTS: At least 1 respiratory tract pathogen was noted in 76 children (96%). Bacteria were found in 73 cases (92%), and viruses were found in 55 (70%). In 52 patients (66%), both bacteria and viruses were found. Bacteria typical of AOM were detected in 86% of patients. Picornaviruses accounted for 60% of all viral findings. CONCLUSIONS: In the great majority of children, AOM is a coinfection with bacteria and viruses. The patent tympanostomy tube does not change the spectrum of causative agents in AOM. A microbiological etiology can be established in practically all cases. PMID- 17083015 TI - Acute otitis media is not a pure bacterial disease. PMID- 17083016 TI - Genetic polymorphism of the binding domain of surfactant protein-A2 increases susceptibility to meningococcal disease. AB - BACKGROUND: Meningococcal disease occurs after colonization of the nasopharynx with Neisseria meningitidis. Surfactant protein (SP)-A and SP-D are pattern recognition molecules of the respiratory tract that activate inflammatory and phagocytic defences after binding to microbial sugars. Variation in the genes of the surfactant proteins affects the expression and function of these molecules. METHODS: Allele frequencies of SP-A1, SP-A2, and SP-D were determined by polymerase chain reaction in 303 patients with microbiologically proven meningococcal disease, including 18 patients who died, and 222 healthy control subjects. RESULTS: Homozygosity of allele 1A1 of SP-A2 increased the risk of meningococcal disease (odds ratio [OR], 7.4; 95% confidence interval [CI], 1.3 42.4); carriage of 1A5 reduced the risk (OR, 0.3; 95% CI, 0.1-0.97). An analysis of the multiple single-nucleotide polymorphisms in SP-A demonstrated that homozygosity for alleles encoding lysine (in 1A1) rather than glutamine (in 1A5) at amino acid 223 in the carbohydrate recognition domain was associated with an increased risk of meningococcal disease (OR, 6.7; 95% CI, 1.4-31.5). Carriage of alleles encoding lysine at residue 223 was found in 61% of patients who died, compared with 35% of those who survived (OR adjusted for age, 2.9; 95% CI, 1.1 7.7). Genetic variation of SP-A1 and SP-D was not associated with meningococcal disease. CONCLUSIONS: Gene polymorphism resulting in the substitution of glutamine with lysine at residue 223 in the carbohydrate recognition domain of SP A2 increases susceptibility to meningococcal disease, as well as the risk of death. PMID- 17083017 TI - Invasive meningococcal disease and a need to understand host genetic susceptibility. PMID- 17083018 TI - Clostridium sordellii infection: epidemiology, clinical findings, and current perspectives on diagnosis and treatment. AB - Clostridium sordellii infections pose difficult clinical challenges and are usually fatal. Most commonly, these infections occur after trauma, childbirth, and routine gynecological procedures, but they have recently been associated with medically induced abortions and injection drug use. We report 2 fatal cases, one of which was associated with minor trauma, and the other of which was associated with normal childbirth, and we summarize the clinical features of 43 additional cases of reported C. sordellii infection. Of these 45 cases, 8 (18%) were associated with normal childbirth, 5 (11%) were associated with medically induced abortion, and 2 (0.4%) were associated with spontaneous abortion. The case fatality rate was 100% in these groups. Ten (22%) of the C. sordellii infections occurred in injection drug users, and 50% of these patients died. Other cases of C. sordellii infection (in 19 patients [43%]) occurred after trauma or surgery, mostly in healthy persons, and 53% these patients died. Overall, the mortality rate was 69% (31 of 45 patients). Eighty-five percent of all patients with fatal cases died within 2-6 days of initial infection, and nearly 80% of fatal cases developed leukemoid reactions. Rapid diagnostic tests and improved treatments are needed to reduced the morbidity and mortality associated with this devastating infection. PMID- 17083019 TI - Clostridium sordellii infection in medical abortion. PMID- 17083020 TI - Issues related to the design and interpretation of clinical trials of salvage therapy for invasive mold infection. AB - Invasive mold infection is a major cause of morbidity and mortality among severely immunocompromised individuals. We discuss the challenges involved in the design and interpretation of salvage antifungal trials, focusing on mold infection. We suggest that patients with refractory fungal infection be analyzed separately from those with intolerance to standard regimens because of the poorer prognosis of the former group. We propose a composite outcome assessment in which refractory infection is defined as infection associated with the worsening of at least 2 of the following 3 types of criteria: clinical, radiologic, and mycologic. Confounding variables, including heterogeneity in host factors, initial antifungal therapy, and selection bias, are discussed. Although randomized studies would provide the most credible results, the lack of an adequate number of patients to meet prespecified stratification criteria for all confounding variables makes such studies impractical. Given that randomized studies are unrealistic, studies involving carefully selected, matched, contemporaneous control subjects are likely to be the most useful alternative. PMID- 17083021 TI - Salvage therapy trials in invasive fungal disease: challenges and opportunities. PMID- 17083022 TI - A patient with dyspnea, cough, and fever. PMID- 17083023 TI - Case studies in cost effectiveness of molecular diagnostics for infectious diseases: pulmonary tuberculosis, enteroviral meningitis, and BK virus nephropathy. AB - Pathogen genome amplification is used to detect and identify microorganisms, assess response to therapy, and detect mutations associated with drug resistance. Nucleic acid amplification tests have been shown to be superior to conventional culture-based testing methods in many circumstances. However, the enthusiasm for the technology in clinical laboratories may be decreased by the practical considerations of cost, complexity of the technology, and lack of US Food and Drug Administration-approved tests. The impact of nucleic acid amplification tests on the diagnosis and management of patients with tuberculosis, enteroviral meningitis, and BK virus transplant nephropathy will be examined, with an emphasis on the potential for health care cost savings. PMID- 17083024 TI - Rifapentine for the treatment of pulmonary tuberculosis. AB - Rifapentine is a recently approved antituberculosis drug that has not yet been widely used in clinical settings. Clinical data support intermittent use of rifapentine with isoniazid during the continuation phase of tuberculosis treatment. Patients with culture-positive, noncavitary, pulmonary tuberculosis whose sputum smear is negative for acid-fast bacilli at the end of the 2-month intensive treatment phase are eligible for rifapentine therapy. Rifapentine should not be used in human immunodeficiency virus-infected patients, given their increased risk of developing rifampin resistance with currently recommended dosages. Rifapentine is not currently recommended for children aged <12 years, pregnant or lactating women, or individuals with culture-negative or extrapulmonary tuberculosis. Rifapentine (600 mg) is administered once weekly with isoniazid (900 mg) during the continuation phase of treatment. This combination should only be given under direct observation. As with rifampin, drug drug interactions are common, and regular patient monitoring is required. Ease of administration makes this regimen attractive both for tuberculosis-control programs and for patients. PMID- 17083025 TI - Women infected with HIV type 1 Brazilian variant, subtype B (B'-GWGR motif) have slower progression to AIDS, compared with patients infected with subtype B (B GPGR motif). AB - INTRODUCTION: The Brazilian variant of human immunodeficiency virus (HIV) type 1 (HIV-1) subtype B (serotype B'-GWGR) has a tryptophan replacing a proline in position 328 of the HIV-1 envelope, a feature that may induce a different HIV disease progression. We aimed to evaluate the role of the B subtypes of HIV-1 (serotypes B-GPGR and B'-GWGR) on HIV disease progression. METHODS: A total of 137 HIV-infected individuals who had been admitted to the hospital were tested with an anti-V3 serologic assay, using peptides representing 2 HIV-1 subtype B strains, MN and SF2, and 2 Brazilian variant B'-GWGR strains, BR1 and BR2. RESULTS: Of 137 serum samples tested with the anti-V3 serologic assay, 4 (3%) yielded indeterminate results, 74 (54%; from 25 women and 49 men) were found to be B-GPGR, and 59 (43%; from 20 women and 39 men) were found to be the B'-GWGR variant. In general, a longer interval from the first known positive HIV test result to an AIDS-defining event was observed in the B'-GWGR group than in the B GPGR group (21 vs. 7 months). The CD4+ T cell counts were higher in the B'-GWGR group (median CD4+ T cell count, 65 vs. 31 cells/mm3; P=.01), and women infected with the B'-GWGR variant were less likely to die than were men infected with the same variant (P=.01). The median viral load in the B'-GWGR group was 3.395 copies/mL, compared with 39.350 copies/mL in the B-GPGR group (P=.01). CONCLUSIONS: Taken together, our results indicate that B'-GWGR-infected women may have more-favorable outcomes than B-GPGR-infected subjects. PMID- 17083026 TI - Risk of cardiovascular disease in a cohort of HIV-infected adults: a study using carotid intima-media thickness and coronary artery calcium score. AB - BACKGROUND: There is concern that human immunodeficiency virus (HIV) infection and the use of highly active antiretroviral therapy lead to accelerated atherosclerosis and increased risk of cardiovascular disease. We measured 2 surrogate markers of subclinical atherosclerosis, carotid intima-media thickness (c-IMT) and coronary artery calcium (CAC) scores, in HIV-infected adults. METHODS: A cross-sectional analysis of 242 men and 85 women with HIV infection was used. Carotid ultrasonography and coronary computed tomography were performed, and their associations with cardiovascular risk factors were examined. RESULTS: Among men, the mean (+/- standard deviation [SD]) common c-IMT was 0.62+/-0.2 mm, the mean (+/-SD) internal c-IMT was 0.76+/-0.5 mm, and 136 patients (56.1%) had detectable CAC. Among women, the mean (+/-SD) common c-IMT was 0.59+/-0.2 mm, the mean (+/-SD) internal c-IMT was 0.66+/-0.4 mm, and 40 patients (47.1%) had detectable CAC. Neither the c-IMT nor the CAC score differed by antiretroviral therapy class or individual medications for either sex. For men, age and waist circumference independently predicted common c-IMT; age, systolic blood pressure, and high-sensitivity C-reactive protein level independently predicted internal c-IMT; and age, apolipoprotein B level, and high sensitivity C-reactive protein level independently predicted CAC score. For women, age and body mass index independently predicted common c-IMT; age independently predicted internal c-IMT; and age and glucose level independently predicted CAC score. CONCLUSIONS: Our participants had more abnormal surrogate markers than expected at a relatively young age, but those were not associated with use of highly active antiretroviral therapy or protease inhibitors. At present, the positive associations were primarily with traditional and novel cardiovascular risk factors. Some HIV-specific (not treatment-specific) factors were observed; they may become more evident with prolonged HIV infection and treatment. PMID- 17083027 TI - An encounter with rabies in New York City. PMID- 17083028 TI - Influenza outbreak among health care workers in an avian influenza (H5N1)-endemic setting. PMID- 17083029 TI - Toward a pharmacogenetic understanding of nucleotide and nucleoside analogue toxicity. PMID- 17083030 TI - Genetic association studies: marking them well. PMID- 17083031 TI - Cotrimoxazole prophylaxis in HIV-infected pregnant women: only a first step. PMID- 17083032 TI - Association between ABCC2 gene haplotypes and tenofovir-induced proximal tubulopathy. AB - BACKGROUND: Tenofovir disoproxil fumarate (TDF) may induce renal proximal tubulopathy (rPT). There are no data on pharmacogenomic predictors of rPT in the genes encoding the multidrug-resistance protein (MRP) 2 and MRP4 transporters. METHODS: Mutational screening of the genes for MRP2 (ABCC2) and MRP4 (ABCC4) was performed using genomic DNA from 13 human immunodeficiency virus type 1 (HIV-1) infected patients (group 1) presenting with TDF-induced rPT. Concomitantly, 17 unrelated HIV-1-infected patients who had received TDF therapy and who did not have rPT (group 2) were included in a case-control analysis, to assess the influence of single-nucleotide polymorphisms (SNPs) identified in ABCC2 and ABCC4. RESULTS: Six SNPs were identified in ABCC2. A significant allelic association between the 1249 G-->A SNP and TDF-induced rPT was observed (odds ratio, 6.11 [95% confidence interval, 1.19-31.15]; P<.02). ABCC2 haplotypes were significantly associated with the onset of TDF-induced rPT--CATC appeared to be a predisposing haplotype, as it was found in 40.9% of the group 1 case patients and in 13.7% of the group 2 control subjects (P<.01), whereas CGAC appeared to be a protective haplotype, as it was not observed in the group 1 case patients but was present in 20.2% of the group 2 control subjects (P<.01). No association was observed between ABCC4 polymorphism and TDF-induced rPT in the present study. CONCLUSION: ABCC2 haplotypes are associated with rPT induced by TDF in HIV-1 infected patients. PMID- 17083033 TI - Exhaustive genotyping of the interleukin-1 family genes and associations with AIDS progression in a French cohort. AB - Interleukin (IL)-1 family members are key players in inflammatory processes but have been the subject of few studies of acquired immunodeficiency syndrome (AIDS). To better evaluate the impact of the IL-1 family on AIDS development, we genotyped the IL1 alpha , IL1 beta , IL1Ra, and IL1R1 genes in 245 slow progressor (SP) and 82 rapid progressor (RP) human immunodeficiency virus type 1 seropositive patients as well as in 446 control subjects, all of whom were of white ethnicity. One hundred sixteen frequent polymorphisms were identified, of which 23 were newly characterized by our study. Many putative associations were found between single-nucleotide polymorphism (SNP) or haplotype alleles and the extreme profiles of progression. Most of them corresponded to weak associations (.01 or =200 cells/ micro L.Conclusion. Antenatal provision of cotrimoxazole for HIV-infected pregnant women with low CD4 cell counts may have indirect benefits for neonatal health. PMID- 17083036 TI - HIV immunosuppression and antimalarial efficacy: sulfadoxine-pyrimethamine for the treatment of uncomplicated malaria in HIV-infected adults in Siaya, Kenya. AB - BACKGROUND: The altered immune response of persons with human immunodeficiency virus (HIV) infection could result in increased rates of antimalarial treatment failure. We investigated the influence of HIV infection on the response to sulfadoxine-pyrimethamine treatment. METHODS: Febrile adults with Plasmodium falciparum parasitemia were treated with sulfadoxine-pyrimethamine and were monitored for 28 days. HIV status and CD4 cell count were determined at study enrollment. RESULTS: Of the adults enrolled in the study, 508 attended all follow up visits, including 130 HIV-uninfected adults, 256 HIV-infected adults with a high CD4 cell count (> or =200 cells/ micro L), and 122 HIV-infected adults with a low CD4 cell count (<200 cells/ micro L). The hazard of treatment failure at day 28 of follow-up was significantly higher for HIV-infected adults with a low CD4 cell count (20.5%) than for HIV-uninfected adults (7.7%). Anemia (hemoglobin level, <110 g/L) modified the effect of HIV status on treatment failure. When we controlled for fever and parasite density, the hazard of treatment failure for HIV-infected adults with a low CD4 cell count and anemia was 3.4 times higher than that for HIV-uninfected adults (adjusted hazard ratio, 3.38; 95% confidence interval, 1.56-7.34). CONCLUSIONS: HIV-infected persons with a low CD4 cell count and anemia have an increased risk of antimalarial treatment failure. The response to malaria treatment in HIV-infected persons must be carefully monitored. Proven measures for the control and prevention of malaria must be incorporated into the basic package of services provided by HIV/acquired immunodeficiency syndrome care and treatment programs in malarious areas. PMID- 17083037 TI - Possible compartmentalization of hepatitis C viral replication in the genital tract of HIV-1-coinfected women. AB - BACKGROUND: We estimated the prevalence of hepatitis C virus (HCV) in cervical cytobrush samples from HCV/human immunodeficiency virus (HIV)-coinfected women and analyzed the HCV quasi species in both cytobrush and plasma samples. Possible compartmentalization of viral quasi species in the genital tract and plasma was evaluated by comparison of genetic heterogeneity and use of phylogenetic analysis. METHODS: Paired plasma and cytobrush samples were obtained from 85 HCV/HIV-coinfected women. The presence of HCV in cytobrush samples was evaluated by reverse-transcription polymerase chain reaction of the 5' untranslated region. Viral quasi species were analyzed by cloning and sequencing the highly variable region-1 in 8 patients. RESULTS: HCV was detected in 27% of cytobrush samples. The composition of viral quasi species was different in the 2 body compartments at both the nucleotide and amino acid level. In fact, the mean complexity was significantly lower in cytobrush samples, and a similar trend was observed for the other parameters of heterogeneity. Phylogenetic analysis and amino acid alignment identified several viral variants that were unique to each body compartment. CONCLUSIONS: Our data suggest that the genital and plasma quasi species represent distinct subpopulations, which possibly reflects compartmentalized viral replication. Alternatively, cell carriers harboring viral quasi species in the genital tract that are distinct from those in plasma could transfer the virus through the barrier separating the 2 body sites. PMID- 17083038 TI - Protective immunity to cytomegalovirus (CMV) retinitis in AIDS is associated with CMV-specific T cells that express interferon- gamma and interleukin-2 and have a CD8+ cell early maturational phenotype. AB - To determine potential correlates of immune recovery from AIDS-related cytomegalovirus retinitis (CMVR), multiparameter flow cytometry was used to characterize CMV-specific T cells from subjects with CMVR. Individuals with active retinitis were compared with those who had been clinically immunorestored by antiretroviral therapy and had > or =2 years of ophthalmologic follow-up without anti-CMV therapy or retinitis reactivation or progression. In comparison with patients with active retinitis, immunorestored patients had higher circulating CD4(+) and CD8(+) T cells expressing interleukin-2 and interferon- gamma in response to combined CMV pp65 and IE1 peptide pool stimulation. CD4(+) T cell responses were predominantly to pp65, whereas CD8(+) T cell responses were predominantly to IE. Immunorestored patients, compared with patients with active retinitis, had increased levels of circulating CMV-specific CD8(+) T cells with "early" (CD27(+)CD28(+)CD45RA(+), CD27(+)CD28(+)CD45RA(-)) and "intermediate" (CD27(-)CD28(+)CD45RA(-)) phenotypes. Recovery from AIDS-related CMVR after the initiation of antiretroviral therapy may be mediated by CMV-specific CD4(+) and CD8(+) T cells capable of promoting antigen-specific CD8(+) T cell proliferation. PMID- 17083039 TI - Pertussis toxin and its binding unit inhibit HIV-1 infection of human cervical tissue and macrophages involving a CD14 pathway. AB - Pertussis toxin (PTX) and its binding unit (PTX-B) have been shown to inhibit human immunodeficiency virus (HIV)-1 infection of primary cells. However, the anti-HIV mechanisms have yet to be defined. We demonstrate that PTX inhibits HIV 1 infection of human cervical tissue independently of viral tropism. PTX-B showed a similar pattern of HIV-1 inhibition. Further investigation in macrophages demonstrated that PTX/PTX-B inhibited HIV-1 expression but that other G protein inhibitors and activators had no effect on HIV-1 replication. Unlike the anti-HIV bacterial lipopolysaccharide, the anti-HIV effects of PTX/PTX-B were not due to beta -chemokine production or coreceptor down-modulation, but they were dependent on interaction with cell-surface receptors. Antibody blocking studies suggested that cell-surface CD14 is very likely to be the principal receptor involved in the anti-HIV effects of PTX/PTX-B. This was further strengthened by the results of surface plasmon resonance analyses. Further definition of the mechanisms of such inhibition may lead to the development of novel HIV-1 prevention strategies. PMID- 17083041 TI - Haplotypes of IL6 and IL10 and susceptibility to human T lymphotropic virus type I infection among children. AB - To characterize a host polygenic profile associated with susceptibility to human T lymphotropic virus type I (HTLV-I) infection, we examined common variants in 11 immune-related genes among Jamaican children born to HTLV-I-seropositive mothers. Compared with HTLV-I seronegatives, haplotypes of IL6 (-660G/-635C/-236G) and IL10 (-6653C/-1116G) were significantly associated with HTLV-I infection in children independent of maternal provirus load and duration of breast-feeding (odds ratio [OR], 4.5 [95% confidence interval {CI}, 1.2-17.6], and OR, 3.5 [95% CI, 1.4-9.0], respectively). Our findings are the first, to our knowledge, to suggest that host variation in both proinflammatory and anti-inflammatory genes could influence susceptibility to HTLV-I infection. PMID- 17083040 TI - Long-term variations in human T lymphotropic virus (HTLV)-I and HTLV-II proviral loads and association with clinical data. AB - BACKGROUND: The human T lymphotropic virus (HTLV)-I or -II proviral load (VL) may be linked to viral pathogenesis, but prospective data on VL and disease outcomes are lacking. METHODS: Using data from a prospective cohort study of HTLV disease outcomes, we examined baseline VLs with real-time quantitative polymerase chain reaction in 122 HTLV-I- and 319 HTLV-II-infected subjects and serial VLs over the course of 6 visits in a subset of 30 HTLV-I- and 30 HTLV-II-infected subjects. Cox and logistic-regression models were used to test baseline associations, and repeated-measures analysis was used to study variations in VL over time. RESULTS: Over the course of a median of 10.4 years, HTLV-I VLs decreased slightly (slope, 0.017 log(10) copies/10(6) peripheral blood mononuclear cells [PBMCs]/year; P=.042) and HTLV-II VLs did not change (slope, -0.019 log(10) copies/10(6) PBMCs/year; P=.165). Changes in VL over time were associated positively with alcohol use (P=.07) and negatively with black race (P=.03) for HTLV-I and positively with smoking (P=.08) for HTLV-II. In the larger group, there was no association between baseline VL and disease outcomes. In the smaller group with serial VL data, there was an association between increasing VL and bladder or kidney infections for both HTLV-I (P=.005) and HTLV-II (P=.022). CONCLUSIONS: HTLV VLs are stable over time, but alcohol and tobacco intake may affect the progression of VLs. The association between increasing VLs and bladder/kidney infection may be explained by early HTLV-related neuropathologic progression. PMID- 17083042 TI - Dengue virus infections in the first 2 years of life and the kinetics of transplacentally transferred dengue neutralizing antibodies in thai children. AB - BACKGROUND: Understanding dengue virus infection in children and the kinetics of maternal dengue neutralizing antibodies is essential for effective dengue immunization of children in endemic areas. METHODS: Serum samples from 219 mother child pairs and 140 children at 3, 6, 9, 12, 18, and 24 months of age from Bangkok, Thailand, were tested for serotype-specific dengue antibodies. Febrile episodes in the children were recorded. RESULTS: Antibodies were found in 97% of cord serum samples and disappeared in 27%, 80%, and 95% of the children by the age of 6, 9, and 12 months, respectively. Geometric mean titers (GMTs) of the antibodies to 4 dengue serotypes decreased to 5.4-15.5 in 6-month-old infants. Eleven of 12 children acquired dengue virus infection at 6 months of age and beyond; 1 had the infection at 3 months of age. Two exhibited undifferentiated febrile illnesses, and 10 had subclinical infections. CONCLUSIONS: Evidence of dengue virus infection and very low GMTs against all dengue serotypes in children at 6 months of age and beyond was demonstrated. There was no evidence that maternal antibodies were harmful to infants. Dengue virus infection rates increase from 12 months of age onward. These data provide information for supporting the optimal age at vaccination. PMID- 17083043 TI - Importance of vertical and horizontal transmission of West Nile virus by Culex pipiens in the Northeastern United States. AB - West Nile virus (WNV) has become established in the northeastern United States, where mosquitoes are inactive during winter. There have been no documented studies to explain how this virus survives winter and reinitiates infection in spring. We report that WNV was vertically transmitted to 2 F(1) female Culex pipiens from a naturally infected female collected in Stratford, Connecticut. One vertically infected F(1) female, which was 168 days old, fed on a hamster that died 8 days later of West Nile disease. This suggests that WNV survives winter in unfed, vertically infected C. pipiens with amplification initiated in spring by horizontal transmission. PMID- 17083044 TI - Lactobacilli expressing variable domain of llama heavy-chain antibody fragments (lactobodies) confer protection against rotavirus-induced diarrhea. AB - BACKGROUND: Rotavirus-induced diarrhea poses a worldwide medical problem in causing substantial morbidity and mortality among children in developing countries. We therefore developed a system for passive immunotherapy in which recombinant lactobacilli constitutively express neutralizing variable domain of llama heavy-chain (VHH) antibody fragments against rotavirus. METHODS: VHH were expressed in Lactobacillus paracasei, in both secreted and cell surface-anchored forms. Electron microscopy was used to investigate the binding efficacy of VHH expressing lactobacilli. To investigate the in vivo function of VHH-expressing lactobacilli, a mouse pup model of rotavirus infection was used. RESULTS: Efficient binding of the VHH antibody fragments to rotavirus was shown by enzyme linked immunosorbent assay and scanning electron microscopy. VHH fragments expressed by lactobacilli conferred a significant reduction in infection in cell cultures. When administered orally, lactobacilli-producing surface-expressed VHH markedly shortened disease duration, severity, and viral load in a mouse model of rotavirus-induced diarrhea when administered both fresh and in a freeze-dried form. CONCLUSIONS: Transformed lactobacilli may form the basis of a novel form of prophylactic treatment against rotavirus infections and other diarrheal diseases. PMID- 17083045 TI - Experimental model to evaluate the human body louse as a vector of plague. AB - Yersinia pestis has been found in human body lice during plague outbreaks. To evaluate the role that the human body louse plays as a vector of plague, we allowed lice to feed on rabbits made bacteremic by intravenous inoculation of 10(9) colony-forming units of 3 strains of Y. pestis. High mortality rates were observed in all lice 2 and 3 days after infection. The lice remained infected with the strains for their life span and excreted viable organisms in their feces from day 1, although they were unable to lay eggs. The lice infected with 2 virulent strains of Y. pestis transmitted the organisms during feeding to uninfected rabbits, which became septicemic and died of plague (with 1 exception) 1 day later. Infections were transmitted to naive lice that were fed on these rabbits, showing that lice can be vectors of Y. pestis in an experimental model. PMID- 17083046 TI - Rapid systemic bone resorption during the course of Staphylococcus aureus-induced arthritis. AB - Bacterial arthritis is the most rapidly progressing destructive joint disease in humans. To evaluate bone resorption and formation, mice were injected with Staphylococcus aureus and killed after 3 and 14 days. Both total and trabecular bone mineral density were, compared with those in uninfected controls, already significantly reduced 3 days after bacterial inoculation. Serum levels of type I collagen fragments were significantly increased and osteocalcin levels decreased in mice infected with S. aureus, compared with those in noninfected mice, 3 and 14 days after bacterial inoculation. This study shows that there is a rapid and easily measurable systemic bone resorption during S. aureus-induced arthritis. PMID- 17083047 TI - The mutant selection window in rabbits infected with Staphylococcus aureus. AB - BACKGROUND: The mutant selection window hypothesis, originally based on agar plate assays, may lead to new antimicrobial dosing strategies that severely restrict the acquisition of resistance. However, it has not been directly tested in an animal model of infection. METHODS: Local infection with Staphylococcus aureus was established in rabbits, and the infected animals were treated orally with various doses of levofloxacin. Changes in levofloxacin concentration, levofloxacin susceptibility, and counts of total and resistant viable bacteria were monitored at the site of infection. RESULTS: S. aureus lost levofloxacin susceptibility when drug concentrations at the site of infection fluctuated between the lower and upper boundaries of the window, defined in vitro as the minimum inhibitory concentration (MIC)(99) and the mutant prevention concentration (MPC), respectively. The upper boundary of the selection window in vivo was estimated as an AUC(24)/MPC value of ~25 h, where AUC(24) is the area under the drug concentration time curve in a 24-h interval. The lower boundary was estimated as an AUC(24)/MIC value of ~20 h. CONCLUSIONS: The mutant selection window exists in vivo, and its boundaries fit well with those determined in vitro. Maintenance of antimicrobial concentrations above the window is expected to suppress the outgrowth of resistant mutant subpopulations. PMID- 17083048 TI - The common gamma chain cytokines interleukin (IL)-2 and IL-7 indirectly modulate blood fluke development via effects on CD4+ T cells. AB - The human pathogen Schistosoma mansoni exhibits a highly evolved and intricate relationship with its host, evading immune destruction while co-opting CD4(+) T cell-driven mechanisms to facilitate parasite development and egg excretion. Because the common gamma ( gamma (c)) chain cytokine interleukin (IL)-7 is also implicated in modulating schistosome development, we investigated whether this effect is mediated indirectly through the essential role that IL-7 plays in CD4(+) T cell growth and survival. We demonstrate that attenuated schistosome development in the absence of IL-7 results from dysregulated T cell homeostasis and not from disruption of direct interactions between schistosomes and IL-7. We also identify an indirect role that another gamma (c) chain cytokine plays in schistosome development, demonstrating that IL-2 expression by CD4(+) T cells is essential for normal parasite development. Thus, cytokines critical for CD4(+) T cell survival and function can mediate indirect but potent effects on developing schistosomes and underscore the importance of CD4(+) T cells in facilitating schistosome development. PMID- 17083049 TI - Lack of Interference between Streptococcus pneumoniae and Staphylococcus aureus in HIV-infected individuals? PMID- 17083052 TI - Naturally acquired immunity to poliovirus: historical observations have been ignored. PMID- 17083053 TI - Influence of high-efficiency particulate air filtration on mortality and fungal infection: a rebuttal. PMID- 17083056 TI - A multinational assessment of complications in type 1 diabetes: the DiaMond substudy of complications (DiaComp) Level 1. AB - The objectives of this study were to describe the global geographic variation of microvascular and macrovascular complications in childhood onset type 1 diabetes (T1D) and to relate any such variation to diabetes care activities such as self blood glucose monitoring and intensive insulin therapy. The DiaComp study is a multinational (17 countries) cross-sectional study of complications in T1D (n=2,657). All participants were diagnosed at < 15 years of age and had a diabetes duration of 524 years when surveyed. Complications were assessed by self report of physician diagnosis. Twenty-two centres in 17 countries achieved at least a 67% response rate and are included in the analyses. Central European centres exhibited high rates of retinopathy (Lithuania=31.6%, Romania=24.2%), laser treatment (Lithuania=25.4%) and neuropathy (Lithuania=29.9%, Romania=12.4%) in those with short duration of diabetes (515 years), as did Cuba for neuropathy (15.4%). For retinopathy the geographic variation in the short-duration group was also pronounced, ranging from 1.6% in Italy to 41.6% in Lithuania, and from 0% in Brazil, Italy and Australia, to 29.9% in Lithuania for laser treatment. Variation was less dramatic for the prevalence of complications in the long-duration group (1525 years). Hypertension and duration were strong consistent predictors of all complications, while women had higher prevalence for half the complications (retinopathy, laser treatment and renal disease). Intensive insulin therapy and self-monitoring of blood glucose showed little association with prevalence of complications. In conclusion, this first population-based account of the geographic variation of T1D complications has demonstrated substantial variation. However, the healthcare practice variables that were measured contributed little toward explaining this variation. PMID- 17083057 TI - Vascular risk reduction in type 2 diabetes evolving treatment paradigms. PMID- 17083058 TI - Reducing residual cardiovascular risk: the relevance of raising high-density lipoprotein cholesterol in patients on cholesterol-lowering treatment. PMID- 17083059 TI - New hypertension guidelines from the National Institute for Health and Clinical Excellence and the British Hypertension Society. AB - A joint initiative between the National Institute for Health and Clinical Excellence (NICE) and the British Hypertension Society (BHS) has led to the publication of new guidelines for the management of hypertension in the community. Recent trial evidence highlighting the increased incidence of new onset diabetes in those exposed to beta-blocker-based treatment regimens, with or without diuretics, compared with those based on calcium channel blockers (CCBs) or angiotensin-converting enzyme (ACE-Is) inhibitors has led to a recommendation that in the uncomplicated patient, beta-blockers are no longer considered suitable options for first-line therapy. Together with mounting evidence that age and ethnicity dictate blood pressure (BP) responsiveness to different classes of antihypertensive drugs, the ACD algorithm is now proposed (formerly ABCD), with ACE inhibitors (ACE-Is) (or angiotensin receptor blockers [ARBs] when ACE-Is are poorly tolerated) preferred in younger patients and CCBs or diuretics preferred for older patients and in black patients of any age. Pathophysiological considerations have influenced proposals for combination therapies with CCBs or diuretics added to ACE-Is in younger patients and vice versa in older patients. Health economic analyses have clearly indicated the cost effectiveness of CCBs which are now elevated to equal standing with diuretics in older patients. PMID- 17083060 TI - From hypertension to heart failure -- are there better primary prevention strategies? AB - Although in the developed world the incidence of and mortality from coronary heart disease (CHD) and stroke have been declining over the last 15 years, heart failure is increasing in incidence, prevalence and overall mortality, despite advances in the diagnosis and management of the condition. Hypertension, alone or in combination with CHD, precedes the development of heart failure in the majority of both men and women. Whilst there have been improvements in the overall management of hypertension, as reflected in rates of diagnosis, awareness, treatment and control of blood pressure (BP), there are still many patients with hypertension who remain undiagnosed or untreated and of those who do receive treatment many fail to achieve current targets for BP control. Placebo controlled trials in hypertension, largely based on diuretic and beta-blocker based regimens, have unequivocally demonstrated that the treatment of hypertension can significantly reduce the incidence of heart failure. Newer treatment strategies offer theoretical and proven practical advantages over established antihypertensive therapy. In particular, AT1-receptor blockers appear to provide benefits beyond BP control and are effective in the treatment of both hypertension and heart failure. Thus, the primary prevention of heart failure in hypertensive patients should be based upon strategies that provide tight and sustained BP control necessitating the use of multiple drugs. However, there is now compelling evidence to suggest that this therapy should include an antihypertensive agent that inhibits the reninangiotensin- aldosterone system (RAAS). PMID- 17083061 TI - Genetic clamping of renin gene expression induces hypertension and elevation of intrarenal Ang II levels of graded severity in Cyp1a1-Ren2 transgenic rats. AB - INTRODUCTION: Transgenic rats with inducible angiotensin II (Ang II)-dependent hypertension (strain name: TGR[Cyp1a1-Ren2]) were generated by inserting the mouse Ren2 renin gene, fused to the cytochrome P450 1a1 (Cyp1a1) promoter, into the genome of the rat. The present study was performed to characterise the changes in plasma and kidney tissue Ang II levels and in renal haemodynamic function in Cyp1a1-Ren2 rats following induction of either slowly developing or malignant hypertension in these transgenic rats. MATERIALS AND METHODS: Arterial blood pressure (BP) and renal haemodynamics and excretory function were measured in pentobarbital sodium-anaesthetised Cyp1a1- Ren2 rats fed a normal diet containing either a low dose (0.15%, w/w for 1415 days) or high dose (0.3%, w/w for 1112 days) of the aryl hydrocarbon indole-3-carbinol (I3C) to induce slowly developing and malignant hypertension, respectively. In parallel experiments, arterial blood samples and kidneys were harvested for measurement of Ang II levels by radioimmunoassay. RESULTS: Dietary I3C increased plasma renin activity (PRA), plasma Ang II levels, and arterial BP in a dose-dependent manner. Induction of different fixed levels of renin gene expression and PRA produced hypertensive phenotypes of varying severity with rats developing either mild or malignant forms of hypertensive disease. Administration of I3C, at a dose of 0.15% (w/w), induced a slowly developing form of hypertension whereas administration of a higher dose (0.3%) induced a more rapidly developing hypertension and the clinical manifestations of malignant hypertension including severe weight loss. Both hypertensive phenotypes were characterised by reduced renal plasma flow, increased filtration fraction, elevated PRA, and increased plasma and intrarenal Ang II levels. These I3C-induced changes in renal haemodynamics, PRA and kidney Ang II levels were more pronounced in Cyp1a1-Ren2 rats with malignant hypertension. Chronic administration of the AT1-receptor antagonist, hypertension, the associated changes in renal haemodynamics, and the augmentation of intrarenal Ang II levels. CONCLUSIONS: Activation of AT1 receptors by Ang II generated as a consequence of induction of the Cyp1a1-Ren2 transgene mediates the increased arterial pressure and the associated reduction of renal haemodynamics and enhancement of intrarenal Ang II levels in hypertensive Cyp1a1-Ren2 transgenic rats. PMID- 17083062 TI - CNP, but not ANP or BNP, relax human isolated subcutaneous resistance arteries by an action involving cyclic GMP and BKCa channels. AB - Natriuretic peptides play an important role in sodium regulation and blood pressure (BP) control. We examined the effects of atrial natriuetic peptide (ANP), brain natriuretic peptide (BNP) and C-type natriuretic peptide (CNP) on human isolated resistance arteries and the mechanisms involved in vasorelaxation. Human subcutaneous resistance arteries were mounted in an isometric myograph and contracted with phenylephrine. CNP, but not ANP or BNP, relaxed arteries in a concentration dependent manner. The action of CNP was unaffected by removal of the endothelium, inhibition of nitric oxide synthase by NG-monomethyl-Larginine or inhibition of soluble guanylate cyclase by 1H-[1,2,4] oxadiazolo [4,3-alpha] quinoxalin-1-one. Blockade of cyclic GMPdependent kinase by 8- bromoguanosine- 3, 5- cyclic monophosphorothioate, Rp-isomer (Rp-8-Br-cGMPS) inhibited CNP relaxation. CNP relaxation was also inhibited by high potassium or iberiotoxin, indicating that it was due to opening of BKCa channels. Omapatrilat, a vasopeptidase inhibitor of neutral endopeptidase and angiotensin-converting enzyme, enhanced the effect of CNP and inhibited responses to Ang I. In summary, CNP, but not ANP or BNP, relaxes human resistance arteries by activating cyclic GMP-dependent kinase and BKCa. The effects of CNP are enhanced by vasopeptidase inhibition and this may contribute to the vasodilator effects of these agents in vivo. Since CNP is widely present in endothelium it may play a role in the regulation of peripheral resistance in man in physiological and pathological circumstances. PMID- 17083063 TI - Influence of gender and genetic variability on plasma angiotensin peptides. AB - INTRODUCTION: We analysed the influence of three polymorphisms of the renin angiotensin system (RAS) (I/D from angiotensin-converting enzyme [ACE], M235T from angiotensinogen gene [ATG] and A1166C from AT1 receptors) on plasma levels of angiotensin I (Ang I), angiotensin II (Ang II) and angiotensin-(1-7) [Ang-(1 7)]. MATERIALS AND METHODS: The study population consisted of a homogeneous group of 93 healthy subjects (43 men and 50 women, mean age: 20.67+/-2.75 years). The mean blood pressure (BP) was 126+/-7/76+/-5 (SD) mmHg and the mean body mass index (BMI) was 22.4+/-2.5 kg/m2. Angiotensin peptides were separated by high performance liquid chromatography (HPLC) and quantified by radio immuno assay (RIA). Genotypes were determined by polymerase chain reaction (PCR) and restriction enzyme analysis. RESULTS: Mean peptide levels were 92.48+/-102.12 pg/ml for Ang I, 22.35+/-10 pg/ml for Ang II, and 31.65+/-27.46 pg/ml for Ang-(1 7). Men had significantly higher levels of Ang-(1-7) (37.76+/-36.47 pg/ml) than women (26.04+/-13.98 pg/ml) (p<0.05). Among genotypes of each polymorphism, men with the T allele showed higher Ang- (1-7) levels compared with those with the MM genotype (p<0.05). Genotype analysis in women showed that higher Ang I levels were related with the DD genotype. When both genders were compared according to genotype, higher values of Ang-(1-7) levels and its molar ratios were found in men, and there was significantly greater Ang I levels in DD genotypes in women than men (136.72+/-112.43 vs . 65.36+/-46.83 pg/mL). CONCLUSIONS: Significant correlations were found between Ang I and Ang II as well as between Ang II and Ang-(1-7) in the different study group distributions. No correlation was found between levels of Ang I and Ang-(1-7). Certain genotypes exert an influence on angiotensin peptide plasma levels which can only be seen when the population is divided according to gender. PMID- 17083064 TI - Effects of ACE inhibition and angiotensin II receptor blockade on glomerular basement membrane protein excretion and charge selectivity in type 2 diabetic patients. AB - Angiotensin-converting enzyme (ACE) inhibitors may reduce urinary albumin excretion (UAE) by decreasing glomerular pressure and increasing glomerular charge selectivity through preservation of glycosaminoglycans. The effect of Angiotensin II antagonism on glomerular charge selectivity remains to be determined. The aim of this study was to compare the effects of an AT1 blocker losartan and an ACE inhibitor (ACE-I) enalapril on UAE, extracellular matrix proteins, glycosaminoglycan excretion (UGAG) and red blood cell anionic charge (RBCCh) which are the indirect markers of glomerular basement membrane anionic content in hypertensive Type 2 diabetic patients. Twenty-four patients were randomised into two groups and received either enalapril (520 mg/d) or losartan (50100 mg/d). All parameters were measured at baseline and after six months of treatment. At the end of six months, systolic and diastolic blood pressures (BP), UAE rates, UGAG excretion and RBCCh were significantly and equally reduced in both treatment groups compared with baseline. RBCCh was negatively correlated with UAE (r=-0.57, p<0.0001) and UGAG excretion (r=-0.57, p<0.0001); UAE was correlated with UGAG excretion (r=0.58, p<0.0001). In conclusion, enalapril and losartan treatment were equally effective in reducing BP, UAE as well as UGAG excretion and preserving RBCCh in hypertensive Type 2 diabetic patients. ACE inhibition and AT1-receptor blockade may have favourable effects on preserving glomerular anionic content in hypertensive diabetic patients. PMID- 17083065 TI - Angiotensin-converting enzyme inhibitors versus angiotensin receptor blockers. A mini-symposium held at the British Hypertension Society Meeting, September 2005. PMID- 17083066 TI - Spotlight on renin: intrarenal renin-angiotensin system--important player of the local milieu. PMID- 17083067 TI - Renin Academy hosts Topical Workshop session at ESH 2006. PMID- 17083068 TI - Role of angiotensin II in cardiovascular disease therapeutic implications of more than a century of research. AB - More than a century of research on the reninangiotensin system (RAS) has uncovered the widespread involvement of angiotensin II (Ang II) in the pathophysiology of cardiovascular diseases. A number of outcomes-based mega trials utilising hard clinical endpoints have revealed beneficial effects of angiotensin receptor antagonists (AIIAs/ARBs) in patients with hypertension, heart failure, diabetic nephropathy, and post-myocardial infarction (MI). The results of these studies not only emphasise the importance of Ang II in the pathophysiology of these diseases but have provided the basis for an evidence based approach for the use of AIIAs in clinical practice. It is hoped that the next 100 years of research into the RAS will uncover hitherto unimaginable therapeutic opportunities. PMID- 17083069 TI - Aldosterone inhibition reduces the risk of sudden cardiac death in patients with heart failure. AB - Sudden cardiac death (SCD) accounts for over 450,000 deaths annually in the USA. Sudden death is estimated to account for approximately 50% of all deaths from cardiovascular causes. Total cardiac mortality has decreased from 728,115 in 1989 to 719,456 in 1999 but the percentage of deaths that are sudden has actually increased from 38% to 47%. Prevention of SCD in patients with risk factors is a high priority for public health professionals. Aldosterone has been shown to be associated with myocardial inflammation, endothelial dysfunction and fibrosis. There is evidence from clinical trials suggesting the role of aldosterone inhibition in prevention of SCD. We reviewed the potential mechanisms and discuss the clinical implications of Randomised ALdactone Evaluation Study (RALES) and Eplerenone Neurohormonal Efficacy and Survival Study (EPHESUS). The use of aldosterone receptor blockers had an overall effect of 21% risk reduction in SCD. Appropriate monitoring and counselling is essential while using either of the aldosterone inhibitors. PMID- 17083070 TI - Aldosterone blockade over and above ACE-inhibitors in patients with coronary artery disease but without heart failure. AB - Recent evidence points to a role for the renin-angiotensin-aldosterone system (RAAS) in the pathogenesis of atherosclerosis and its complications, including acute angina pectoris. Two large trials in heart failure have clearly demonstrated that blocking aldosterone improves mortality and that this benefit occurs over and above standard therapy with angiotensin-converting enzyme (ACE) inhibitors. The question that naturally arises from these landmark studies is whether aldosterone blockade would produce the same benefits in patients with coronary artery disease (CAD) but no heart failure. There are three reasons to believe this might be the case. Firstly, angiotensin II (Ang II) and aldosterone produce similar biological effects and Ang II withdrawal has been shown to benefit patients with angina; aldosterone blockade may therefore follow in the footsteps of ACE inhibitors, as it did in heart failure, and produce benefits in vascular patients without heart failure. Secondly, one of the main mechanisms which is thought to be responsible for the benefit of aldosterone blockade in the Randomised ALdactone Evaluation Study (RALES) and Eplerenone Post-AMI Heart Failure Survival Study (EPHESUS), is that it improves endothelial/vascular function and endothelial/vascular dysfunction is the fundamental abnormality in angina pectoris. Finally, aldosterone blockade has been shown to reduce atherosclerosis in animal studies of atherosclerosis without heart failure, which are analogous to CAD patients. PMID- 17083071 TI - Differences between cardiac and arterial fibrosis and stiffness in aldosterone salt rats: effect of eplerenone. AB - BACKGROUND: Previous experiments have studied separately the development of either cardiac or aortic fibrosis and stiffness in aldosterone (Aldo)-salt hypertensive rats. Our aim was to determine in vivo the effects of Aldo and the Aldo receptor antagonist eplerenone (Epl) on simultaneous changes in cardiac and arterial structure and function and their interactions. METHODS AND RESULTS: Aldo was administered in uninephrectomised Sprague-Dawley rats receiving a high-salt diet from 8 to 12 weeks of age. Three groups of Aldo-salt rats were treated with 1 to 100 mg/kg-1. d-1 Epl by gavage. Arterial elasticity was measured by elastic modulus (Einc)-wall stress curves using medial cross-sectional area (MCSA). The cardiac and arterial walls were analysed by histomorphometry (elastin and collagen), immunohistochemistry (EIIIA fibronectin, Fn), and Northern blot (collagens I and III). Aldo caused increased systolic blood pressure (SBP), carotid Einc, MCSA, and EIIIA Fn with no change in wall stress or elastin and collagen densities. No difference in collagen mRNA levels was detected between groups. During the same period, cardiac mass and collagen mRNA and protein levels increased markedly in the myocardial tissue. Epl normalised collagen in the myocardium, Eincwall stress curves, MCSA, and EIIIA Fn in Aldo rats. These dose dependent effects were not accompanied by a consistent reduction in SBP and cardiac mass. CONCLUSIONS: In exogenous hyperaldosteronism in the rat, Aldo causes independently myocardial collagen and arterial Fn accumulation, the latter being responsible for increased intrinsic carotid stiffness. Epl prevents both cardiac and arterial effects but does not reduce consistently SBP. PMID- 17083072 TI - Beneficial effect of eplerenone on cardiac remodelling and electrical properties of the failing heart. AB - INTRODUCTION: The effect of chronic administration of eplerenone on cardiac remodelling and electrical properties was investigated in the failing heart of cardiomyopathic hamsters (TO-2) at five months of age. MATERIALS AND METHODS: Two month-old hamsters were treated with eplerenone (200 mg/kg/day) administered into the chow for a period of three months. Measurements of membrane potential were performed with intracellular microelectrodes connected to a high impedance DC amplifier. The thickness of the ventricular wall as well as the area of fibrosis were measured. To investigate the influence of eplerenone on the electrogenic sodium pump myocytes were isolated from the ventricle and the pump current density was measured in voltage clamped cells using the whole cell clamp configuration. RESULTS: The results indicated that: 1) the width of the left and right ventricular wall was significantly reduced; 2) the heart weight/body weight ratio was decreased by 38+/-2.4% (n=24) (p<0.05); 3) the fibrotic area in the left ventricle (LV) was reduced by 12.6+/-2% (n=25) (p<0.05); 4) the incidence of cardiac arrhythmias was decreased from 58+/-3.8% (n=20) in the control to 40+/ 4.1% (n=20) (p<0.05) in animals treated with eplerenone. Moreover, a significant reduction in the dispersion of the QT interval was found with the drug; 5) eplerenone increased the resting potential of ventricular fibres from 64.3+/-1.5 mV to 73.4+/-1.4 mV (n=30) (p<0.05), an effect related to the activation of an electrogenic sodium pump. The conduction velocity, in longitudinal direction, was enhanced from 50+/-2.2 cm/s (n=10) in the controls to 59+/-2.4 cm/s (n=13) (p<0.05) in animals treated with eplerenone. CONCLUSIONS: Eplerenone reduces cardiac remodelling, the incidence of cardiac arrhythmias and improves impulse propagation, an effect in part related to the antifibrotic effect of the drug but also to the activation of the electrogenic sodium pump. PMID- 17083073 TI - Renal haemodynamics are not related to genotypes in offspring of parents with essential hypertension. AB - INTRODUCTION. The pathogenesis of essential hypertension (EH) has a major genetic component and is associated with renal abnormalities. Normotensive offspring of hypertensive parents are likely to develop EH and are a suitable population for identifying possible relations between genetic and renal abnormalities. METHODS: We investigated if renin-angiotensinaldosterone system associated genotypes (angiotensinogen [M235T] and ACE [I/D]) are related to blood pressure (BP), renal haemodynamics and sodium excretion in sex and age-matched (1835 years) healthy Caucasian offspring of either two parents with EH (n=101, EH-offspring) or two normotensive parents (n=50, controls). The alpha-adducin polymorphism (G460W) was also investigated. RESULTS: Compared to controls, BP, heart rate, renal vascular resistance (RVR) and urinary sodium excretion were, respectively, 5%, 7%, 15% and 20% higher in EHoffspring. In controls, the TT-genotype of the M235T angiotensinogen polymorphism was associated with higher BP and higher plasma angiotensinogen. By contrast, in EHoffspring the TT-genotype was associated with lower BP and unchanged plasma angiotensinogen. Plasma angiotensinogen correlated positively with BP in EH-offspring, with a similar tendency (p=0.08) in controls. The distributions of the three candidate polymorphisms were similar in EH offspring and controls. There were no associations between any of the polymorphisms and any of the renal parameters measured. CONCLUSION: The markedly greater RVR, proportionally larger than the greater BP, supports a role for RVR in the pathogenesis of EH. The lack of association between the candidate polymorphisms and the investigated parameters, even in this homogenous and for hypertension strongly predisposed group, suggests that the polymorphisms investigated do not play important roles in the pathogenesis of hypertension. PMID- 17083074 TI - Spotlight on renin: cardiovascular outcomes in chronic kidney disease and suppression of the renin-angiotensin system. PMID- 17083085 TI - Articular cartilage collagen: an irreplaceable framework? AB - Adult articular cartilage by dry weight is two-thirds collagen. The collagen has a unique molecular phenotype. The nascent type II collagen fibril is a heteropolymer, with collagen IX molecules covalently linked to the surface and collagen XI forming the filamentous template of the fibril as a whole. The functions of collagens IX and XI in the heteropolymer are far from clear but, evidently, they are critically important since mutations in COLIX and COLXI genes can result in chondrodysplasia syndromes. Here we review what is known of the collagen assembly and present new evidence that collagen type III becomes covalently added to the polymeric fabric of adult human articular cartilage, perhaps as part of a matrix repair or remodelling process. PMID- 17083086 TI - Multitrack electrospray chips. AB - Multitrack electrospray chips (MTEC) were fabricated by UV-photoablation of polyethylene terephthalate (PET) substrates. They are composed of an array of up to six microchannels that are successively used as electrospray ionization (ESI) emitters for mass spectrometry (MS). There is no requirement for alignment of the different spraying microchannels with the mass spectrometer orifice. The MTEC is thus fixed in front of the mass spectrometer and the successive MS analyses are performed without moving the chip. The sequential electrospraying by successive application of an identical high voltage in each off-axis microchannel was evaluated for the fast screening of peptides and proteins. The counting of cysteines in peptides through chemical modification and the relative quantification of a peptide in two samples are presented herein as two original strategies based on this new analytical tool. PMID- 17083087 TI - MRI in fetal necropsy. AB - The fetal autopsy involves a series of investigations of the corpse, most of which are noninvasive and acceptable to the majority of parents and their physicians. The value of the perinatal autopsy is manyfold and well established, and the results can provide a basis for parental and family counseling, inform future obstetric management, and provide audit for prenatal care. Many techniques originally developed for diagnosis, such as histology, biochemical tests, photography, x-rays, and cytogenetic karyotyping, have become standard tools in perinatal autopsies. However, there has been an inexorable decline in the autopsy consent rate over the last 30 years due to social and cultural factors, and perhaps ignorance of the benefits to be derived from the examination. Growing evidence suggests that postmortem fetal MRI can assist the pathologist at autopsy, and in many cases can obviate the need for dissection or at least minimize and focus it. For the majority of cases in which no consent for surgical autopsy is given, MRI together with other noninvasive postmortem tests can provide a great deal of the information that was previously available only from autopsy. PMID- 17083088 TI - Insight into the activity of SARS main protease: Molecular dynamics study of dimeric and monomeric form of enzyme. AB - The phenomenon that SARS coronavirus main protease (SARS M(pro)) dimer is the main functional form has been confirmed by experiment. However, because of the absence of structural information of the monomer, the reasons for this remain unknown. To investigate it, two molecular dynamics (MD) simulations in water for dimer and monomer models have been carried out, using the crystal structure of protomer A of the dimer as the starting structure for the monomer. During the MD simulation of dimer, three interest phenomena of protomer A have been observed: (i) the distance between NE2 of His41 and SG of Cys145 averages 3.72 A, which agrees well with the experimental observations made by X-ray crystallography; (ii) His163 and Glu166 form the "tooth" conformational properties, resulting in the specificity for glutamine at substrate P1 site; and (iii) the substrate binding pocket formed by loop 140-146 and loop 184-197 is large enough to accommodate the substrate analog. However, during the MD simulation of the monomer complex, the three structural characteristics are all absent, which results directly in the inactivation of the monomer. Throughout the MD simulation of the dimer, the N-terminus of protomer B forms stable hydrogen bonds with Phe140 and Glu166, through which His163, Glu166, and loop 140-146 are kept active form. Furthermore, a water-bridge has been found between the N-terminus of protomer B and Gly170, which stabilizes His172 and avoids it moving toward Tyr161 to disrupt the H-bond between Tyr161 and His163, stabilizing the conformation of His163. The interactions between the N-terminus and another monomer maintain the activity of dimer. PMID- 17083089 TI - Spatial and spectral heterogeneity of time-varying shear stress profiles in the carotid bifurcation by phase-contrast MRI. AB - PURPOSE: To determine the heterogeneity of the time-varying shear stress profiles in the human carotid bifurcation, a region prone to atherosclerosis. MATERIALS AND METHODS: Lagrangian bicubic interpolation of phase-contrast MRI images was used to determine the shear stress profiles for three adult healthy male volunteers. Frequency spectra for the common and internal carotid artery (CCA and ICA, respectively)-derived shear stresses were examined in order to determine the presence of significant heterogeneity in the intensity distribution. RESULTS: Hemodynamic characteristics (peak, minimum, average shear stress, and oscillatory shear index [OSI]) were highly heterogeneous both along the length of the vessel as well as circumferentially around the CCA and ICA. In the frequency domain, intensities below 4 Hz were significantly higher in the CCA compared to the sinus region of the ICA, indicating that shear stress heterogeneity can be detected in the frequency domain. The harmonic index, a measure of the relative contributions of dynamic and static components of the shear stress signal, colocalizes with OSI, which implies a relationship between specific frequency components and atherosclerosis development. CONCLUSION: These findings indicate that the time and frequency dependent parameters of in vivo shear stress have important implications for regional development of atherosclerosis. PMID- 17083090 TI - Use of the U-clip for microvascular anastomosis. AB - Microvascular anastomosis is a demanding skill requiring technical excellence and a thorough knowledge of anatomy and physiology. Every suture placed in a microvascular anastomosis should be considered critical as each has the potential to compromise the delicate reconstruction. As such, any device that can facilitate microvascular suture placement deserves thorough evaluation. The U clip (Coalescent Surgical, Sunnyvale, CA) is such a device in that it eliminates the often time consuming process of tying knots. We evaluated use of the U-clip in microvascular anastomosis of a 1.5 mm artery. We found the U-clip to offer some advantages including ease of use, traditional feel of directed suture placement (as compared to couplers), and elimination of time needed for knot tying. Its shortcomings include size (in diameter, the "pop-off" section of the device appears larger than standard 8-0 suture), the significant force required to "pop-off" the device and difficulty removing the device. PMID- 17083091 TI - Pharmacokinetics of tamoxifen after intravenous and oral dosing of tamoxifen hydroxybutenyl-beta-cyclodextrin formulations. AB - Oral and intravenous administration of tamoxifen base and tamoxifen citrate formulated with hydroxybutenyl-beta-cyclodextrin (HBenBCD) to Sprague-Dawley rats significantly increased the oral bioavailability of tamoxifen relative to that of parent drug (no HBenBCD). When formulated with HBenBCD, the form of tamoxifen (base vs. salt) made no difference in the oral bioavailability of tamoxifen. Liquid formulations (PG:PEG400:H2O) provided higher oral bioavailability than solid formulations dissolved and dosed as aqueous oral solutions. The oral bioavailability of tamoxifen was significantly influenced by both dietary status and time of dosing of the animals. Tamoxifen metabolite plasma concentrations were not affected by complexation of tamoxifen with HBenBCD. Collectively, the data indicated that dosing of fasted animals in the morning with tamoxifen:HBenBCD formulations provided a very significant increase in tamoxifen oral bioavailability (up to 10- to 14-fold). PMID- 17083092 TI - Drug discarding: a problematic therapeutic event in Parkinson's disease. PMID- 17083093 TI - Differentiation of hepatocellular carcinoma and hepatic metastasis from cysts and hemangiomas with calculated T2 relaxation times and the T1/T2 relaxation times ratio. AB - PURPOSE: To determine the diagnostic capability of the T1 and T2 relaxation times and the T1/T2 relaxation times ratio generated with the mixed turbo spin echo (mixed-TSE) pulse sequence, in order to discriminate between hepatocellular carcinoma (HCC)/metastases and hemangiomas/cysts. MATERIALS AND METHODS: A retrospective review of 36 MR examinations implementing the mixed-TSE pulse sequence demonstrated 70 focal hepatic lesions. Quantitative MR algorithms were used to generate T1 and T2 relaxation times, and the T1/T2 relaxation times ratio for each lesion. A two-sample t-test compared mean T1 and T2 relaxation times, and the T1/T2 relaxation times ratio, by lesion type: carcinoma/metastases and hemangiomas/cysts. Sensitivity and specificity for discriminating carcinoma/metastases from hemangiomas/cysts with T2 relaxation time thresholds of 112 and 125 msec, as well as a ratio of T1/T2 relaxation times of 5.8, were calculated. RESULTS: Using a T2 relaxation time threshold of 112 msec, 92% sensitivity and 100% specificity discriminating cysts/hemangiomas from HCC/liver metastasis was demonstrated. With a threshold of 125 msec, 96% sensitivity and 98% specificity was demonstrated. There was no correlation between calculated T1 relaxation times and type of lesion. Using a T1/T2 relaxation times ratio of 5.8, 100% sensitivity and specificity were demonstrated. CONCLUSION: Although there is high sensitivity and specificity associated with the use of T2 relaxation times alone to discriminate carcinoma/metastases from hemangiomas/cysts, using the T1/T2 relaxation times ratio threshold of 5.8 allowed proper classification of all lesions. PMID- 17083094 TI - Highly concentrated monoclonal antibody solutions: direct analysis of physical structure and thermal stability. AB - Virtually all current analytical methods employed in the development of highly concentrated monoclonal antibody (MAb) formulations require dilution of the sample before acquiring data. Thus, there is an unmet need for methods to study proteins directly at high concentration, since extrapolation of stability indicating parameters obtained from dilute studies may not be representative of the high concentration solution. Only slight or no modifications of biophysical methods including fluorescence, UV absorbance, circular dichroism, and FTIR (ATR and transmittance) spectroscopies as well as differential scanning calorimetry (DSC) are described here that permit the direct study of immunoglobulins (and other proteins) at high concentrations. Although FTIR spectra show differences that are dependent upon sampling geometry, other spectroscopic data from two different recombinant MAbs suggests that structure of each antibody exists in a physically similar state in the concentration range of 0.1-190 mg/mL in 40 mM pH 6 citrate-phosphate buffer. Upon thermally stressing these proteins, spectroscopic techniques that probe tertiary structure demonstrate a decrease in the apparent thermal melting temperature of approximately 5-20 degrees C of both proteins with increasing concentration. In contrast, DSC thermograms and CD thermal experiments suggest a minor degree of stabilization (approximately 2 degrees C) for both antibodies although protein association could be responsible for these observations. Empirical phase diagrams produced from spectroscopic data also suggest (1) the existence of similar structural states at low temperatures independent of concentration and (2) a decrease in the temperature at which phase changes are observed with increasing concentration. The decrease in structural stability observed in these studies is probably the result of aggregation or self association of the recombinant MAbs upon heating in crowded solutions and not due to a decrease in the intrinsic structural stability of the MAbs. PMID- 17083095 TI - Dopaminergic innervation of the human striatum in Parkinson's disease. PMID- 17083096 TI - Transcranial brain sonography findings in clinical subgroups of idiopathic Parkinson's disease. AB - To investigate whether transcranial brain sonography (TCS) discriminates different courses of idiopathic Parkinson's disease (PD), 101 patients with clinically definite PD were studied. In four patients, TCS was not possible due to insufficient acoustic temporal bone windows. Substantia nigra (SN) hyperechogenicity was found in 96% of assessable patients. Larger SN echogenic size correlated with younger age at PD onset (Spearman correlation, r = -0.383; P < 0.001), but not with age, PD duration, or severity. Marked bilateral SN hyperechogenicity indicated early-onset rather than late-onset PD, and akinetic rigid (AR) or mixed-type (MX) PD rather than tremor-dominant PD. SN echogenic sizes were larger contralateral to the clinically more affected side in AR PD and MX PD patients. Reduced echogenicity of brainstem raphe was associated with depression (RR = 1.61; 95% CI = 1.05-2.46; P = 0.044) but not with other clinical features. Caudate nucleus hyperechogenicity was, independently from PD duration, related to drug-induced psychosis (RR = 2.40; CI = 1.36-4.22; P = 0.001), but not to motor fluctuations. Lenticular nucleus hyperechogenicity indicated AR PD rather than tremor-dominant PD (RR = 1.44; CI = 1.11-1.86; P = 0.040). Frontal horn dilatation > 15.4 mm (mean of bilateral measurements) indicated increased risk of dementia (RR = 4.11; CI = 1.51-11.2; P = 0.001). We conclude that TCS displays characteristic changes of deep brain structures in different clinical manifestations of PD. PMID- 17083097 TI - Management of refractory urinary urge incontinence following urogynecological surgery with sacral neuromodulation. AB - AIMS: We sought to explore our patient outcomes utilizing sacral neuromodulation in the management of refractory urinary urge incontinence following urogynecological surgical procedures. METHODS: A total of 25 women with urinary urge incontinence following urogynecological surgery were selected for SNS therapy and retrospectively analyzed. All patients completed a comprehensive urological evaluation. Clinical data was recorded to determine outcomes and identify parameters that would be predictive of response to neuromodulation. Outcomes were determined via subjective patient questionnaire and graded as follows: significant response (> or =80% improvement), moderate response (> or =50% and <80% improvement), and poor response (<50% response). RESULTS: Nineteen patients had a previous pubovaginal sling (10 with concomitant pelvic prolapse repair), 3 a previous retropubic suspension, and 3 a transperitoneal vesicovaginal fistula repair. Urethrolysis was performed in 4 patients to alleviate bladder outlet obstruction prior to sacral neuromodulation. Mean patient age was 59.8 years and length of follow-up was 7.2 months. Twenty-two women (88%) had the IPG placed during a Stage 2 procedure. Twenty patients maintained at least a 50% improvement in clinical symptoms at last follow-up and 6 patients were continent. Overall, the number of pads/day improved from 4.2 to 1.1 (P < 0.001). There were no significant differences in response to neuromodulation based upon age, duration of symptoms, type of surgery, or urodynamic parameters. CONCLUSION: Sacral neuromodulation appears to be an effective therapy in patients with refractory urge incontinence following urogynecological surgery. Larger prospective studies with longer follow-up are needed to assess the durability of this therapeutic modality. PMID- 17083098 TI - Revisiting safety of minocycline as neuroprotection in Huntington's disease. PMID- 17083099 TI - Patient with late-onset Wilson's disease: deterioration with penicillamine. PMID- 17083100 TI - Transobturator tape (TOT): Two years follow-up. AB - AIMS: The aim of this study is to report the functional results, patient satisfaction, and morbidity of the Transobturator tape procedure (TOT) in the treatment of stress incontinence (SUI). METHODS: One hundred and thirty patients were prospectively evaluated with history, physical examination, quality of life questionnaire including Incontinence Impact Questionnaire (IIQ), urogenital distress inventory (UDI), and analog global satisfaction scale (GSS), and urodynamic studies. RESULTS: One hundred and seventeen patients (90%) had history of SUI, and 78 (60%) had urge incontinence. Pads/day (PPD) used was 2.48 +/- 2.42, and the score of IIQ 16.13 +/- 7.86, UDI 10.95 +/- 3.4, and GSS 1.41 +/- 1.67. All patients underwent TOT using the ObTape. Hospital stay was 0.84 +/- 0.76 days and catheter duration was 1.42 +/- 2.08 days. At a follow-up of 16.85 +/- 4.68 months, 13 patients (10%) have recurrent SUI, 21 (16.15%) persistent urge incontinence, and 1 (1.92%) de novo urge incontinence. The mean PPD is 0.15 +/- 0.56, IIQ 1.47 +/- 5.14, UDI 3.28 +/- 3.09, and GSS 8.29 +/- 1.64. Two patients (1.52%) developed urethral obstruction, five (3.84%) had vaginal extrusion of the tape, and two (1.52%) had intra-operative bladder perforation. CONCLUSIONS: These results demonstrate the safety and efficacy of the TOT. The short hospitalization and catheterization, low incidence of de novo urge incontinence and obstructive voiding offers a distinct advantage over existing techniques. No significant difference in outcome between patients with VLPP < or =60 cm H2O, and patients with VLPP >60 cm H2O was observed. PMID- 17083101 TI - Bilateral hemifacial spasm and trigeminal neuralgia: a unique form of painful tic convulsif. PMID- 17083102 TI - LRRK2 G2019S founder haplotype in the Chinese population. AB - The G2019S mutation in the LRRK2 (leucine-rich repeat kinase) gene appears very rarely in the Chinese population. Among Chinese subjects who were non-G2019S carriers, we demonstrated the frequency of the LRRK2 G2019S founder haplotype (T 254-A-G-A-154) in Parkinson's disease and controls to be 33% and 30%. This rate is similar to the frequency in European noncarriers, indirectly supporting the association of this haplotype with G2019S carriers. The haplotype is likely to be more ancient than the G2019S mutation because it is also found in a population with a very low carrier rate. PMID- 17083103 TI - Statistical representation of mean diffusivity and fractional anisotropy brain maps of normal subjects. AB - PURPOSE: To create diffusion tensor atlases from echo planar imaging (EPI) images acquired at 3 T in 10 normal subjects. MATERIALS AND METHODS: Data from 10 right handed healthy adult volunteers (mean age of 31 +/- 3 years; eight males) were acquired using a 3.0-T scanner. Geometric distortion artifacts correction was accomplished by combining parallel acquisition to reduce the distortion as well as postprocessing by registration to a geometrically accurate T2-weighted fast spin-echo image. This reduced distortions to within a voxel for most of the internal structures of the brain. The apparent diffusion coefficient (ADC) and fractional anisotropy (FA) atlases were created by warping images using an iterative optical-flow-based local deformation algorithm that used two channels of data: ADC and FA. RESULTS: A three-dimensional distance measure was used to evaluate the accuracy of the registration algorithm with contours defined on two structures: the corpus callosum and cerebellum. The average three-dimensional distance value for the nine subjects (with the 10th as the reference) was 0.2 mm for the corpus callosum and 1.2 mm for the cerebellum. CONCLUSION: A high resolution, diffusion MR atlas with full brain coverage was developed. Additionally, maps of the SD of the diffusion indices were also generated to provide an estimate of the variance within a normal population. Active shape and texture models were also generated for the corpus callosum as an alternate method of representing the variance in morphology and diffusion indices. PMID- 17083104 TI - Extradural motor cortex stimulation in Parkinson's disease. AB - Extradural motor cortex stimulation (EMCS) is a surgical procedure proposed for patients with advanced Parkinson's disease (PD) who cannot undergo deep brain stimulation (DBS). Five PD patients with motor fluctuations and dyskinesia underwent EMCS of the left hemisphere. All fulfilled CAPSIT criteria for DBS, with the exception of age > 70 years. Patients were assessed preoperatively and 6 months after surgery on and off medications, with stimulator on, and 2 weeks later with stimulator off. Outcome measures included changes in mean medication dosage (levodopa and dopamine agonists), Unified Parkinson's Disease Rating Scale (UPDRS Parts II-III and Item 39), and dyskinesias (Abnormal Involuntary Movements Scale [AIMS]). We found no significant mean changes following EMCS. However, there was a trend for a reduction of mean daily medication intake (-30%) and AIMS (-19%). There were 3 patients who reported reduced OFF time (UPDRS Item 39) and 4 of 5 who felt a subjective benefit in stability and gait. In our PD cohort, EMCS induced no objective benefit, although some subjective improvement was reported mostly on axial symptoms. PMID- 17083105 TI - Broad-Complex, E74, and E75 early genes control DNA puff BhC4-1 expression in prepupal salivary glands. AB - The DNA puff BhC4-1 gene of the sciarid Bradysia hygida is induced in salivary glands prior to the pupal molt as a secondary response to the increase in ecdysone titers. Previous studies demonstrated that the BhC4-1 promoter is activated in transgenic Drosophila melanogaster salivary glands as a late response to the ecdysone peak that triggers metamorphosis, revealing that this aspect of BhC4-1 transcriptional regulation is conserved in the Drosophila background. To identify regulators of BhC4-1 expression, we utilized a candidate gene approach and tested the roles of the ecdysone-induced genes BR-C, E74, and E75. Our results reveal that the BR-C Z3 isoform is essential for BhC4-1-lacZ induction in prepupal salivary glands and constitute the first demonstration of the participation of early genes products on DNA puff genes regulation. PMID- 17083106 TI - Minimally invasive parathyroidectomy without intraoperative parathyroid hormone monitoring in patients with primary hyperparathyroidism. AB - BACKGROUND: Minimally invasive parathyroidectomy (MIP) is the preferred operation for patients with primary hyperparathyroidism (HPT) and positive preoperative imaging. This non-randomized case series assessed the long-term results of MIP performed without the use of intraoperative parathyroid hormone (ioPTH) monitoring. METHODS: The study involved prospective collection of demographic, biochemical and operative details on a consecutive, unselected cohort of 298 patients who underwent surgery for non-familial primary HPT during a 5-year interval. The mean preoperative serum calcium level was 3.00 mmol/l with a mean parathyroid hormone concentration of 25.8 pmol/l. (99m)Tc-labelled sestamibi scanning and neck ultrasonography were performed in 262 patients. RESULTS: Sestamibi scan showed unilateral uptake in 182 patients and a single parathyroid adenoma was confirmed on ultrasonography in 161 patients. MIP was performed in 150 patients. The mean duration of operation was 25 (range 8-65) min. Four patients needed conversion to conventional neck exploration. There was one postoperative haematoma and three cases of temporary recurrent laryngeal nerve neuropraxia. All but four patients were normocalcaemic after MIP. All the parathyroid tumours removed were adenomas, with a mean weight of 1.3 (range 0.1 17.4) g. No patient developed recurrent HPT after a median follow-up of 16 (range 3-48) months. CONCLUSION: The outcome of MIP without ioPTH monitoring was comparable to that reported in series that used ioPTH monitoring. PMID- 17083107 TI - Impact of radiotherapy on local recurrence of rectal cancer in Norway. AB - BACKGROUND: The purpose of this study was to analyse the impact of radiotherapy on local recurrence of rectal cancer in Norway after the national implementation of total mesorectal excision (TME). METHODS: This was a prospective national cohort study of 4113 patients undergoing major resection of rectal carcinoma between November 1993 and December 2001. RESULTS: The proportion of patients who had radiotherapy before or after operation increased from 4.6 per cent in 1994 to 23.0 per cent in 2001. The cumulative 5-year local recurrence rate decreased from 16.2 to 10.7 per cent. Multivariable analysis showed that preoperative radiotherapy significantly reduced local recurrence (hazard ratio 0.59 (95 per cent confidence interval 0.39 to 0.87)). The use of preoperative radiotherapy in patients from a local hospital offering radiotherapy was 50 per cent higher than that for patients from a hospital without such services (P = 0.003); cumulative 5 year local recurrence rates for these patients were 10.6 and 15.8 per cent respectively (P < 0.001). CONCLUSION: Following national implementation of TME for rectal cancer, increased use of preoperative radiotherapy appeared to reduce recurrence rates further. PMID- 17083108 TI - MRI to assess arrhythmia and cardiomyopathies. AB - Cardiovascular magnetic resonance (CMR) is highly precise for morphological and functional analyses of the myocardium, and has been used to assess different types of cardiomyopathies. Its ability to characterize tissue, especially with gadolinium (Gd) delayed-enhancement techniques, has shown promising results for the diagnosis of arrhythmogenic cardiomyopathies. In this review we discuss the background and potential of this approach, as well as its usefulness for assessing arrhythmias and cardiomyopathies. PMID- 17083109 TI - Recombinase-mediated cassette exchange to rapidly and efficiently generate mice with human cardiac sodium channels. AB - SCN5A encodes the predominant voltage-gated sodium channel isoform in human heart and nearly 100 variants have now been described and studied in vitro. However, development of animal models to analyze function of such large numbers of human gene variants represents a continuing challenge in translational medicine. Here, we describe the implementation of a two stage procedure, recombinase-mediated cassette exchange (RMCE), to efficiently and rapidly generate mice in which a full-length human cDNA replaces expression of the murine ortholog. In the first step of RMCE, conventional homologous recombination in mouse ES cells was used to replace scn5a exon 2 (that contains the translation start site) with a cassette acceptor that includes the thymidine kinase gene, flanked by loxP/inverted loxP sites. In the second step, the cassette acceptor site was replaced by the full length wild-type human SCN5A cDNA by Cre/loxP-mediated recombination. The exchange event occurred in 7/29 (24%) colonies, and the time from electroporation to first homozygotes was only 8 months. PCR-restriction fragment length polymorphism (RFLP) showed that the murine isoform was replaced by the human one, and functional studies indicated that mice with human cardiac sodium channels have wild-type sodium current density, action potential durations, heart rates, and QRS durations. These data demonstrate that RMCE can be used to generate mice in which a targeted allele can be rapidly and efficiently replaced by variants of choice, and thereby can serve as an enabling approach for the functional characterization of ion channel and other DNA variants. PMID- 17083110 TI - Magnetic resonance investigation of bone marrow following priming and stem cell mobilization. AB - PURPOSE: To evaluate application of MRI and magnetic resonance spectroscopy (MRS) to monitor bone marrow cellularity during pretransplant priming with chemotherapy and hematopoietic growth factor (HGF) administration. MATERIALS AND METHODS: A total of 10 lymphoma and myeloma patients, in remission following induction therapy and considered eligible for high-dose therapy and autologous stem cell transplantation, were included in the study. MR investigation was scheduled four times: at study entry, and one, two, and four weeks following priming. Priming with cyclophosphamide and recombinant human granulocyte colony-stimulating factor (rhG-CSF) started the day after study entry. MR parameters studied in a region of interest were as follows: bone marrow intensity on short-time inversion-recovery (STIR) turbo spin-echo (TSE; thus STIRTSE) and on T1-weighted TSE (T1TSE) images, T2 value for fat component, T2 value for water component, water/fat ratio (W/F), T1 value for fat component, and T1 value for water component. RESULTS: The results did not support the hypothesis that hematopoietic expansion quantitated and monitored by MR correlates to the level of mobilized progenitor cells. CONCLUSION: The results indicate that release of stem cells is a more complex phenomenon than hematopoietic expansion and reduction of fat tissue in bone marrow. PMID- 17083111 TI - Effectiveness of MR imaging for diagnosing the mild forms of acute pancreatitis: comparison with MDCT. AB - PURPOSE: To assess the usefulness of MRI and determine which MR sequence is most effective for diagnosing the mild forms of acute pancreatitis. MATERIALS AND METHODS: Forty subjects (20 normal volunteers and 20 patients with a mild form of acute pancreatitis) underwent MRI with fat-suppressed T1-weighted fast low-angle shot (FLASH), half-Fourier acquisition single-shot turbo spin-echo (HASTE), and TSE short TI inversion recovery (TSE-STIR) sequences. The 20 patients with a mild form of acute pancreatitis underwent multidetector CT (MDCT) and MRI within a 24 hour interval. We qualitatively analyzed all of the images by assessing inflammatory changes in the pancreas and peripancreatic fat. We quantitatively compared differences in pancreas-liver contrast between the control and patient groups for each MR sequence by measuring the signal intensities of the pancreas and liver. RESULTS: TSE-STIR was the best of the four modalities for delineating peripancreatic and pancreatic inflammation (P < 0.01). TSE-STIR depicted definitive peripancreatic and pancreatic inflammation in 18 and 15 patients, respectively. MDCT depicted only three cases of peripancreatic inflammation. TSE STIR was also produced the best the best quantitative results of the MR sequences (P = 0.09). CONCLUSION: MRI is helpful for diagnosing the mild forms of acute pancreatitis. We recommend the use of TSE-STIR imaging as part of the routine protocol for evaluating pancreatitis. PMID- 17083112 TI - Esophageal carcinoma: evaluation with high-resolution three-dimensional constructive interference in steady state MR imaging in vitro. AB - PURPOSE: To determine the usefulness of high-resolution three-dimensional (3D) constructive interference in steady state (CISS) MRI for evaluating mural invasion and morphologic features in esophageal carcinomas. MATERIALS AND METHODS: Twenty-four esophageal specimens with carcinomas were studied with a 1.5 T system using a 4-cm-diameter loop coil. High-resolution 3D-CISS MR images were obtained with a field of view (FOV) of 80 mm, matrix of 256 x 256, and section thickness of 0.5 mm (voxel size of 0.05 mm(3)). 3D-CISS MR images were compared with histopathologic findings, and virtual MR endoscopic images were compared with macroscopic findings at surgery. RESULTS: 3D-CISS MR images clearly depicted the normal esophageal wall as consisting of eight layers, which correlated well with the histologic layers. In 22 of 24 esophageal carcinomas (92%), the depth of mural invasion visualized with 3D-CISS MRI correlated well with the histopathologic staging. In all 24 carcinomas (100%), virtual MR endoscopic images clearly depicted the macroscopic types of the carcinomas, including adjacent lymph node swelling. CONCLUSION: High-resolution 3D-CISS MRI has a high diagnostic accuracy for evaluating mural invasion and macroscopic findings in esophageal carcinomas, and may be applicable to preoperative histopathologic staging and morphologic evaluation. PMID- 17083113 TI - Temporospatially regulated expression of subtilisin-like proprotein convertase PACE4 (SPC4) during development of the rat submandibular gland. AB - The temporospatial expression of PACE4, a member of the mammalian subtilisin-like proprotein convertase family involved in the activation of growth/differentiation factors, was investigated by in situ hybridization during the development of the rat submandibular gland (SMG). At the initiation stage (day 15.5 of gestation; E15), PACE4 was intensely expressed in the submandibular epithelium, but weakly expressed in the mesenchymal cells. At E16 when the branching morphogenesis becomes obvious, the expression of PACE4 in the mesenchyme was further decreased, although its level in the submandibular epithelium had not changed remarkably from that at E15. During the next stage of embryonic development (E17-E20), PACE4 was expressed in the cells derived from the submandibular epithelium, which include the proacinar, terminal tubular, and presumptive ductal cells. In the perinatal SMG, PACE4 was still expressed intensely in the terminal portion of the SMG containing the proacinar and terminal tubular cells, whereas its expression in the ductal cells was obviously decreased at the second postnatal day (P2) and at P6. Acinar cells expressing no PACE4 appeared, and their numbers increased following their development (P9-P20). At P30 when the PACE4 expression in the acinar cells was completely suppressed, its expression in the ductal cells became intense again. This temporospatially regulated expression of PACE4 suggests its apparent association with the proliferation, differentiation, and establishment of functional acinar and ductal cells of the SMG. PMID- 17083116 TI - Biocompatible superparamagnetic iron oxide nanoparticle dispersions stabilized with poly(ethylene glycol)-oligo(aspartic acid) hybrids. AB - Methoxypoly(ethylene glycol)-oligo(aspartic acid) (MPEG-Asp(n)-NH(2), n = 2-5) hybrid block copolymers were synthesized and used as stabilizers to prepare superparamagnetic Fe(3)O(4) nanoparticles with magnetite as the inner core and and poly(ethylene glycol) as the hydrophilic outer shell. The aqueous dispersions of the nanoparticles were stable at pH 2-11 and in 1M NaCl solution, when repeat number, n, was 3 or more. Transmission electron microscopy showed that the nanoparticles, stabilized with MPEG-Asp(3)-NH(2), were about 14 nm in diameter. Magnetic measurements indicated that MPEG-Asp(3)-NH(2)-coated iron oxide nanoparticles showed superparamagnetic behavior. Cell adhesion assay and in vitro cell viability/cytotoxicity studies showed that MPEG-Asp(3)-NH(2)-coated iron oxide nanoparticles had less effect on cell adhesion/viability and morphology, and less cytotoxicity compared with uncoated, poly (acrylic acid)-coated, and MPEG-poly(acrylic acid)-coated iron oxide nanoparticles. PMID- 17083117 TI - Validation of the short forms of the incontinence impact questionnaire (IIQ-7) and the urogenital distress inventory (UDI-6) in a Turkish population. AB - AIM: To validate the Turkish versions of the IIQ-7 and UDI-6 for use in Turkish speaking populations. METHODS: After establishing the test-retest reliability and internal consistency in a pilot study, 302 patients were enrolled in the study and general and subscale scores of the questionnaires were calculated. All participants underwent an urodynamic assessment. RESULTS: Both instruments showed a high internal consistency (Cronbach's alpha for the IIQ-7 and UDI-6 was 0.87 and 0.74, respectively) and test-retest reliability (Spearman's rho was 0.99 for both of the scales (P < 0.001). 55.6% of the participants showed urodynamic abnormality and/or leakage. 39.7% had urodynamic stress incontinence (USI) and 15.9% had detrussor overactivity (DOA) +/- USI. The mean scores of each IIQ-7 and UDI-6 were significantly higher in the USI, and DOA +/- USI groups compared with normal women. Women with DOA +/- USI scored highest in the IIQ-7 and UDI-6. The irritative subscale scores of the 1st and 2nd items of the UDI-6 were significantly higher in the DOA +/- USI group. The stress subscale scores of 3rd and 4th items of UDI-6 were significantly higher in the USI group. Women with postvoid residual (PVR) urine values greater than 50 ml had significantly higher obstructive subscale scores compared to the ones who had less residual volumes. CONCLUSIONS: The Turkish translated versions of the IIQ-7 and UDI-6 are reliable, consistent and valid instruments for assessing symptom severity and the impact on QOL in Turkish speaking women with urinary incontinence. PMID- 17083118 TI - Honey, I shrunk the DNA: DNA length as a probe for nucleic-acid enzyme activity. AB - The replication, recombination, and repair of DNA are processes essential for the maintenance of genomic information and require the activity of numerous enzymes that catalyze the polymerization or digestion of DNA. This review will discuss how differences in elastic properties between single- and double-stranded DNA can be used as a probe to study the dynamics of these enzymes at the single-molecule level. PMID- 17083119 TI - In vivo MR imaging of pulmonary arteries of normal and experimental emboli in small animals. AB - PURPOSE: To demonstrate the feasibility of pulmonary MRA in living rodents. MATERIALS AND METHODS: A three-dimensional (3D) gradient echo sequence was adapted to perform a time-of-flight (TOF) angiography of rat lung. Angiogram with a spatial resolution of 195 x 228 x 228 microm(3) was acquired in around 33 minutes. The method was then applied in animals before and after pulmonary embolism (PE) induction. Section of the proximal right pulmonary artery was measured and compared between the two populations. RESULTS: Good quality images were obtained with a contrast-to-noise ratio (CNR) of 9 +/- 3 in the proximal part of the pulmonary artery. Cross-section areas of the right main artery are statistically different before (3.45 +/- 0.69 mm(2)) and after induction of PE (4.3 +/- 0.86 mm(2)). CONCLUSION: This noninvasive tool permits angiogram acquisition at around 200 microm spatial resolution and objective distinction between healthy and embolized arteries. PMID- 17083120 TI - Implanted deep brain stimulator and 1.0-Tesla magnetic resonance imaging. AB - There is a great need for MRI examinations of patients who have previously undergone deep brain stimulator (DBS) implantation. The current guidelines pertain only to a 1.5-Tesla horizontal-bore scanner complying with strict safety regulations. Moreover, almost all published in vitro and in vivo studies concerning patient safety are carried out on 1.5 Tesla MR scanners. The aim of our work is to share our clinical experience of 1.0-Tesla brain MR imaging. During the past four years, 34 patients with different types of implanted DBS systems underwent 1.0-Tesla MR examinations to answer diagnostic or clinical questions. Apart from the scanner type applied, all other safety instructions were strictly followed. The MRI itself made no significant difference to the measured impedances or the stimulation parameters required to achieve the optimal therapeutic results. From theoretical considerations, it may be assumed that 1.0 Tesla MRI can be performed safely on DBS-implanted patients, provided that all other recommendations are adhered to. PMID- 17083121 TI - Projections of somatosensory cortex and frontal eye fields onto incertotectal neurons in the cat. AB - The goal of this study was to determine whether the input-output characteristics of the zona incerta (ZI) are appropriate for it to serve as a conduit for cortical control over saccade-related activity in the superior colliculus. The study utilized the neuronal tracers wheat germ agglutinin-horseradish peroxidase (WGA-HRP) and biotinylated dextran amine (BDA) in the cat. Injections of WGA-HRP into primary somatosensory cortex (SI) revealed sparse, widespread nontopographic projections throughout ZI. In addition, region-specific areas of more intense termination were present in ventral ZI, although strict topography was not observed. In comparison, the frontal eye fields (FEF) also projected sparsely throughout ZI, but terminated more heavily, medially, along the border between the two sublaminae. Furthermore, retrogradely labeled incertocortical neurons were observed in both experiments. The relationship of these two cortical projections to incertotectal cells was also directly examined by retrogradely labeling incertotectal cells with WGA-HRP in animals that had also received cortical BDA injections. Labeled axonal arbors from both SI and FEF had thin, sparsely branched axons with numerous en passant boutons. They formed numerous close associations with the somata and dendrites of WGA-HRP-labeled incertotectal cells. In summary, these results indicate that both sensory and motor cortical inputs to ZI display similar morphologies and distributions. In addition, both display close associations with incertotectal cells, suggesting direct synaptic contact. From these data, we conclude that inputs from somatosensory and FEF cortex both play a role in controlling gaze-related activity in the superior colliculus by way of the inhibitory incertotectal projection. PMID- 17083122 TI - Mice lacking the p75 receptor fail to acquire a normal complement of taste buds and geniculate ganglion neurons by adulthood. AB - Brain-derived neurotrophic factor and neurotrophin-4 are required for normal taste bud development. Although these neurotrophins normally function via the tyrosine kinase receptor, trkB, they also bind to the pan-neurotrophin receptor, p75. The goal of the present study was to determine whether the p75 receptor is required for the development or maintenance of a full complement of adult taste buds. Mice with p75 null mutations lose 34% of their circumvallate taste buds, 36% of their fungiform papillae, and 26% of their fungiform taste buds by adulthood. The reduction of taste buds in the adult circumvallate papilla was similar to that observed previously at postnatal day 7 (Fan et al. Brain Res Dev Brain Res 2004;150:23-39). Taken together, these findings indicate that the p75 receptor is critical for the development of a full complement of taste buds, but is not required for maintenance of circumvallate taste buds in adulthood. Immunolabeling for p75 was not observed in taste buds, indicating that p75 signaling influences taste bud number indirectly. Geniculate ganglion neurons, which provides innervation to fungiform taste buds, express the p75 receptor. Mice with p75 null mutations also have fewer neurons in the geniculate ganglion. Together, these results suggest that the p75 receptor is important for the survival of geniculate neurons and geniculate neuron survival is required for the development of a full complement of taste buds by adulthood. PMID- 17083123 TI - A cohort study of thyroid cancer and other thyroid diseases after the Chornobyl accident: pathology analysis of thyroid cancer cases in Ukraine detected during the first screening (1998-2000). AB - BACKGROUND: The Ukrainian American Cohort Study evaluated the risk of thyroid disorders in a group of individuals who were younger than age 18 years at the time of the Chornobyl (Chernobyl) accident. In this article, the authors describe the pathology of thyroid carcinomas detected in the first screening. METHODS: From 1998 to 2000, 13,243 individuals completed the first cycle of screening examinations. Eighty patients underwent surgery between 1998 and 2004. Intraoperative and postoperative pathologic studies were performed at the Institute of Endocrinology and Metabolism, Kyiv. RESULTS: Pathologic analysis revealed 45 thyroid carcinomas, including 43 papillary thyroid carcinomas (PTCs) (95.6%) and 2 follicular thyroid carcinomas (FTCs) (4.4%). TNM classification (5th edition) of the PTCs included 8 T1 tumors (18.6%), 16 T2 tumors (37.2%), and 19 T4 tumors (44.2%). Fifteen PTCs (34.9%) were N1a,N1b, and 3 PTCs (7.0%) were M1. Among the PTCs, 8 exhibited the classical papillary histologic pattern (18.6%), 14 exhibited a follicular histologic pattern (32.6%), 5 exhibited a solid histologic pattern (11.6%), and 16 exhibited a mixed histologic pattern (37.2%). Both FTCs had a microfollicular-solid structure. Eleven of 20 cohort members who underwent surgery before the first screening had PTCs. Regional metastases (63.6%) and distant metastases (18.2%) were more common in this group. CONCLUSIONS: Multifocal growth, lymphatic and blood vessel invasion, extrathyroid spread, and regional and distant metastases were more frequent in less differentiated PTCs (>30% solid structure). Small carcinomas (80 (P < .01) and a hemoglobin (Hgb)level at presentation of >/=12 (P = .03). On multivariate analysis, KPS was the only independent prognostic factor for disease free survival. Median disease-free survival was 4.9 months among patients with a KPS score of >80 and was 3.9 months among those with a KPS score of /=12 (P = .02), KPS>80 (P < .001), and <5% weight loss (P = .03). On multivariate analysis, Hgb and KPS were independent prognostic factors for overall survival. CONCLUSIONS: In the current study, KPS score was an independent prognostic factor for disease-free and overall survival among patients treated with chemoradiation for LAPC. The pretreatment Hgb level was an additional independent prognostic factor for overall survival. PMID- 17083125 TI - Loss of neutral endopeptidase and activation of protein kinase B (Akt) is associated with prostate cancer progression. AB - BACKGROUND: Neutral endopeptidase (NEP) is a cell-surface peptidase that can regulate the activation of Akt kinase through catalytic-dependent and independent mechanisms. NEP expression is absent in approximately 50% of prostate cancers. The authors investigated whether NEP loss in vivo would result in Akt phosphorylation and potentially contribute to prostate cancer progression by examining the interaction of NEP, Akt, and phosphatase and tensin homolog (PTEN) in a prostate xenograft model and in clinical specimens from patients with prostate cancer. METHODS: Using a tetracycline-repressible expression system to express NEP in a tumor animal xenograft model, the effects of NEP were tested on tumor growth, Akt phosphorylation, and PTEN expression. The clinical relevance of NEP, phosphorylated Akt, and PTEN protein expression also was investigated in 204 patients who had undergone radical prostatectomy. RESULTS: The results indicated that the induction of NEP expression inhibited established xenograft tumor growth, diminished Akt phosphorylation, and increased PTEN protein levels. In humans, prostate cancers with complete loss of NEP expression were significantly more likely to express phosphorylated Akt (P = .02). Moreover, patients who had prostate cancers with concomitant loss of NEP and expression of phosphorylated Akt had an increased, independent risk of prostate-specific antigen (PSA) recurrence (P = .03). In the study cohort, loss of PTEN protein expression did not correlated significantly with phosphorylated Akt or with patients' clinical outcome. CONCLUSIONS: The findings from this investigation demonstrated that NEP loss leads to Akt activation and contributes to the clinical progression of prostate cancer. PMID- 17083126 TI - Treatment outcome for metastatic papillary renal cell carcinoma patients. AB - BACKGROUND: Most clinical trial reports in metastatic renal cell carcinoma (RCC) do not distinguish between histologic subtypes, making it difficult to assess specific treatment efficacy. The current retrospective study sought to define clinical features and outcome data for metastatic papillary RCC. METHODS: Clinical features, treatment outcome, and survival were evaluated in 38 patients with metastatic papillary RCC who underwent clinical evaluation at Memorial Sloan Kettering Cancer Center (MSKCC) between 1985 and 2005. Twenty-three of 513 individuals were identified from a clinical trial database, 14 of 1895 from a surgery database, and 1 of 357 from a pathology database. A literature review of systemic therapy in metastatic papillary RCC was performed. RESULTS: Among the 38 patients, 30 had been treated at MSKCC with various systemic therapies, including cytokines. Twelve therapies resulted in stable disease, 30 in initial progression of disease, and 1 in an unknown response. One patient had a partial response to sunitinib, a novel multitargeted tyrosine kinase inhibitor. The median overall survival time for the entire study group was 8 months (95% confidence interval, 5 12). A literature review on treatment of metastatic papillary RCC produced 4 reports, confirming a lack of efficacy for systemic therapy. CONCLUSIONS: A resistance to systemic therapy characterizes patients with metastatic papillary RCC. Further understanding of the genetics and molecular biology and subtypes involved may provide the basis for more effective agents. Treatment with targeted therapies or other experimental agents is warranted. PMID- 17083128 TI - Parathyroid hormone related protein (PTHrP) inhibits TNFalpha-induced apoptosis by blocking the extrinsic and intrinsic pathways. AB - Parathyroid hormone related protein (PTHrP) is expressed at low levels in many fetal and adult tissues where it plays a central role in regulating cell proliferation, cell death, and tissue homeostasis. In vivo and in vitro, PTHrP has been shown to promote the survival of a variety of cells by regulating expression of the anti-apoptotic protein Bcl2. Additional work has shown that intra-nuclear accumulation of PTHrP in CFK2 (PTH1R positive) and 27m21 (PTH1R negative) condrogenic cells promotes their survival by closing down ribosome biogenesis and promoting quiescence. The current studies were undertaken to examine the role of wild-type PTHrP and a mutant form that cannot translocate to the nucleus in protecting cells from TNFalpha-induced apoptosis. Both forms of the protein were equally effective in blocking the extrinsic pathway by inhibiting expression of the TNF receptor death domain, activating Bid, and promoting cleavage of caspase 8. These observations suggest a direct mechanism of PTHrP action on components of the extrinsic pathway, involving a region of the protein outside of the NTS. PTHrP and M1PTHrP also inhibited the intrinsic pathway by preventing the exchange of anti-apoptotic for pro-apoptotic proteins at the mitochondrial membrane, thus maintaining its integrity and preventing the release of caspase-activating factors into the cytosol. In general, this mitochondrial-related activity was somewhat delayed and was mediated more effectively by PTHrP than by M1PTHrP, suggesting an indirect mechanism of action that might require the presence of an intact NTS. PMID- 17083129 TI - Regulation of translation initiation by the yeast eIF4E binding proteins is required for the pseudohyphal response. AB - The eukaryotic translation initiation factor eIF4E is responsible for the recognition of the mRNA cap structure and, as such, plays a key role in the selection of mRNAs for translation. The interaction of eIF4E with the 'multi adaptor' eIF4G (and thus recruitment of ribosomes to mRNA) can be regulated via competitive binding of 4E-binding proteins (4E-BPs). 4E-BPs have broad functions in cell growth, proliferation and development. We have found that disruption of the genes for either of the yeast 4E-BPs (Eap1p or Caf20p) leads to an inhibition of pseudohyphal growth in the resulting diploid yeast strain following nitrogen limitation. Specific 4E-binding domain mutations destroy the capacity of each 4E BP gene to complement the non-pseudohyphal phenotype, suggesting that a translational function for the 4E-BPs is important for pseudohyphal growth. In addition, neither of the 4E-BP deletion strains is deficient in global or stress regulated protein synthesis. However, our evidence reveals that the two 4E-BPs are functionally distinct with regard to pseudohyphal growth. Therefore, this work supports a model where the yeast 4E-BPs are acting on specific mRNAs to facilitate a defined proliferative response to environmental stress in yeast. PMID- 17083130 TI - Psychotropics use in the Spanish elderly: predictors and evolution between years 1993 and 2003. AB - PURPOSE: This study mainly aimed at describing the prevalence of psychotropic medication consumption in the Spanish elderly population between the years 1993 and 2003. METHODS: Descriptive, cross-sectional study covering the Spanish population aged 65 years and over, using data drawn from the 1993 and 2003 Spanish National Health Surveys (ENSS). A total of 9570 interviews were analysed (3436 from 1993 and 6134 from 2003). The independent variables were sociodemographic and health-related, and the dependent variable was total consumption of psychotropic medication. Using logistic multivariate regression models, we have analysed the temporal evolution of psychotropic medication consumption between 1993 and 2003. RESULTS: The prevalence of consumption was significantly higher in women (6.7% from 1993 and 26.4 % from 2003) versus men (2.4% from 1993 and 10.6% from 2003) (p < 0.001). Multivariate analysis, highlighted the association between increased psychoactive drug intake and sex, nervous, depressive, sleep disorders and negative perception of health, displayed a strong association with consumption of psychoactive drugs across the 2 years. CONCLUSIONS: In Spain, the prevalence of psychoactive drug consumption is higher among elderly women than men, and increases with negative perception of health. The prevalence of consumption was significantly higher in the year 2003. PMID- 17083131 TI - Solid-phase microextraction method for carbon isotopic analysis of volatile carboxylic acids in human plasma by gas chromatography/combustion/isotope ratio mass spectrometry. AB - A new analytical method is described for the determination of the physiological concentration and low-level enrichment of (13)C-short-chain volatile organic acids (SCVAs) (e.g. (13)C-acetate and (13)C-butyrate) in human plasma. This two step method involves solid-phase microextraction (SPME) coupled to gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) without any organic solvents or derivatizing agents. Two SCVA extraction methods were compared using a carboxen/polydimethylsiloxane fiber: headspace sampling (HS) and liquid sampling (LS) SPME. The influences of extraction temperature and time were tested to optimize the adsorption of SCVAs onto the fiber. The comparison of the peak area responses of the acids in the two adsorption methods showed better sensitivity in the human physiological concentration range in the LS mode than in the HS mode. The accuracy of isotopic enrichment measurement was determined using plasma spiked with (13)C-acetate and (13)C-butyrate solution from 0 to 1 mol percent excess (MPE). The linearity and repeatability (RSD < 5%) were measured in LS mode. Plasma SCVA concentrations were also determined relative to 3 methylvalerate (internal standard). Linearity and repeatability were observed from 0 to 400 microM for acetate, from 0 to 20 microM for propionate, and from 0 to 10 microM for butyrate. This method was also used to determine plasma acetate production obtained from lactulose (an undigestible disaccharide) fermentation in one healthy volunteer over 3 h. The acetate concentration increased twofold, 2 h after oral lactulose intake. These results are in agreement with the data obtained by GC/MS in healthy volunteers and obese adults following a lactulose intake by using higher amounts of labelled tracers.SPME coupled with GC/C/IRMS can be used to analyze (13)C-SCVAs at low enrichment (<0.5 MPE) within the physiological concentration measured in human plasma. PMID- 17083132 TI - Identification of the novel thio-arsenosugars DMThioAsSugarCarboxyl, DMThioAsSugarCarbamate and DMThioAsSugarAdenine in extracts of giant clam tissues by high-performance liquid chromatography online with electrospray tandem mass spectrometry. AB - Recently, the identification of four thio-arsenosugars in extracts of marine samples was reported. The biosynthesis of these arsenic species is most likely closely related to the well known oxo-arsenosugars. Due to their symbiotic algae giant clams contain oxo-arsenosugars with Carboxyl-, Carbamate- and Adenine aglycone in significant amounts. Now we provide evidence for the presence of the thio analogues of these three arsenosugars in tissue extracts of the giant clams Tridacna maxima and Tridacna derasa. Anion-exchange high-performance liquid chromatography (HPLC) online with electrospray tandem mass spectrometry was applied for the identification of the thio-arsenosugars and the indirect quantification via their oxo analogues. PMID- 17083133 TI - Combining near infrared spectroscopy and multivariate analysis as a tool to differentiate different strains of Saccharomyces cerevisiae: a metabolomic study. AB - Near-infrared (NIR) spectroscopy has gained wide acceptance within the food and agriculture industries as a rapid analytical tool. NIR spectroscopy offers the advantage of rapid, non-destructive analysis and routine operation is simple and opens the possibility of using spectra to obtain the 'fingerprint' of a sample. The aim of this study was to explore the potential of combining visible (VIS) and near-infrared (NIR) spectroscopy, together with multivariate analysis, in establishing the function of genes, by investigating the metabolic profiles produced by Saccharomyces cerevisiae deletion strains sourced from the EUROSCARF yeast collection. Spectra (400-2500 nm) were acquired with a FOSS NIRSystems6500 (Foss NIRSystems), in transmittance mode. Principal component analysis (PCA) and linear discriminant analysis (LDA) were used in order to visualize graphically the relative differences and similarities of yeast deletion strains. VIS and NIR spectroscopy showed great promise as a screening tool for both discriminating between yeast strains and grouping strains with deletions in genes that disturb similar metabolic pathways. These results indicate that the methods may be useful in defining the function of genes that produce no obvious phenotype. PMID- 17083134 TI - Systematic hybrid LOH: a new method to reduce false positives and negatives during screening of yeast gene deletion libraries. AB - We have developed a new method, systematic hybrid loss of heterozygosity, to facilitate genomic screens utilizing the yeast gene deletion library. Screening is performed using hybrid diploid strains produced through mating the library haploids with strains from a different genetic background, to minimize the contribution of unpredicted recessive genetic factors present in the individual library strains. We utilize a set of strains where each contains a conditional centromere construct on one of the 16 yeast chromosomes that allows the destabilization and selectable loss of that chromosome. After mating a library gene deletion haploid to such a conditional centromere strain, which corresponds to the chromosome carrying the gene deletion, loss of heterozygosity (LOH) at the gene deletion locus can be generated in these otherwise hybrid diploids. The use of hybrid diploid strains permits complementation of any spurious recessive mutations in the library strain, facilitating attribution of the observed phenotype to the documented gene deletion and dramatically reducing false positive results commonly obtained in library screens. The systematic hybrid LOH method can be applied to virtually any screen utilizing the yeast non-essential gene deletion library and is particularly useful for screens requiring the introduction of a genetic assay into the library strains. PMID- 17083135 TI - New vectors for simplified construction of BrdU-Incorporating strains of Saccharomyces cerevisiae. AB - The thymidine analogue BrdU is a powerful tool for analysing nucleotide incorporation in studies of DNA replication or repair. S. cerevisiae lacks the thymidine salvage pathway that enables efficient cellular uptake and incorporation of thymidine analogues into DNA. Recent in vivo reconstitution of this pathway in yeast by high-level expression of Herpes simplex virus thymidine kinase (HSV-TK) or combined expression of HSV-TK and human equilibrative nucleoside transporter (hENT1) has enabled analysis of BrdU incorporation in yeast. While the BrdU incorporation systems are highly valuable, the construction and use of strains utilizing these systems can be complicated by specific requirements of the available systems. We have created a set of vectors that simplify the construction and use of BrdU-Incorporating (BrdU-Inc) strains of budding yeast. Each vector in the set contains HSV-TK and hENT1 under the control of promoters that express constitutively, and one of four different selectable markers (HIS3, TRP1, LEU2 or URA3) for genomic integration. With these BrdU-Inc vectors, one-step integration of a single copy produces yeast that efficiently incorporate BrdU upon its addition to the medium. These vectors ease strain construction and maintenance, thereby facilitating routine use of BrdU for analysis in yeast. PMID- 17083136 TI - Yeast Derlin Dfm1 interacts with Cdc48 and functions in ER homeostasis. AB - Recent studies have identified Derlin-1, a protein that associates with the AAA ATPase p97 and is implicated in late steps in ER-associated protein degradation (ERAD). Derlin-1 has two Saccharomyces cerevisiae homologues, Der1p and Dfm1p. While Der1p has been studied extensively, little is known about Dfm1p. Accordingly, we investigated the role of Dfm1p in ERAD, ER homeostasis and interactions with the yeast p97 homologue Cdc48p. Dfm1p was not involved in the degradation of a number of Der1-dependent or -independent ERAD substrates, neither was it redundant with either Der1p or Sec61p in ERAD. However, Dfm1p had a role in ER homeostasis, since Dfm1p loss or overexpression could stimulate the unfolded protein response (UPR). Furthermore, Dfm1p interacted both genetically and physically with Cdc48p, the yeast p97 homologue, and this interaction required an eight amino acid sequence found in the C-terminus of Dfm1p that we have termed the SHP box. Our genetic studies are consistent with the lack of a role for Dfm1p in ERAD, but indicate it participates in ER-related Cdc48p actions distinct from retrotranslocation. Finally, sequence analysis indicated that the UPR-related and Cdc48p interaction functions of Dfm1p could be separated, implying this protein probably has numerous actions in the cell. Thus, the interaction between Derlins and p97 is conserved between yeast and mammals, although its function in ERAD is not. Furthermore, Dfm1p interacts with Cdc48p through its SHP boxes, and so defines a new motif for interaction with this widely-employed AAA-ATPase. PMID- 17083137 TI - Control of pi-electron rotation in chiral aromatic molecules by nonhelical laser pulses. PMID- 17083138 TI - Controllable [2+2] cycloadditions of 1,5-bisallenyl-substituted compounds. PMID- 17083139 TI - A new non-centrosymmetric modification of BiB(3)O(6). PMID- 17083140 TI - Linker-controlled energy and charge transfer within chlorophyll trefoils. PMID- 17083141 TI - Stereoselective synthesis of pyrrolidines: catalytic oxidative cyclizations mediated by osmium. PMID- 17083143 TI - Microfluidics device for single cell gene expression analysis in Saccharomyces cerevisiae. AB - We have measured single-cell gene expression over time using a microfluidics based flow cell which physically traps individual yeast using microm-sized structures (yeast jails). Our goal was to determine variability of gene expression within a cell over time, as well as variability between individual cells. In our flow cell system, yeast jails are fabricated out of PDMS and gene expression is visualized using fluorescently-tagged proteins of interest. Previously, single-cell yeast work has been done using micromanipulation on agar, or FACS. In the present device agar is eliminated, resulting in a superior optical system. The flow of media through the flow cell washes daughter cells away, eliminating the need for micromanipulation. Unlike FACS, the described device can track individual yeast over a time course of many hours. The flow cells are compatible with the needs of quantitative fluorescence microscopy, and allow simultaneous measurements to be done on a large number of individual yeast. We used these flow cells to determine the expression of HSP104-GFPand RAS2-YFP, genes known to affect yeast life span. The results demonstrate inter-cell variation in expression of both genes that could not have been detected without this single-cell analysis. PMID- 17083144 TI - Fetal growth patterns in fetuses of women with pregestational diabetes mellitus. AB - OBJECTIVE: To assess the effect of glucose control on the rate of growth of fetuses in women with pregestational diabetes mellitus (Types 1 and 2). METHODS: All pregestational diabetic women booked at Mater Mothers' Hospital, Brisbane, Australia, between 1 January 1994 and 31 December 2002, were included. Pregnancies with congenital fetal anomalies, multiple pregnancies, and pregnancies terminated prior to 20 weeks' gestation were excluded. Dating scans were performed before 14 weeks' gestation and serial scans were performed at 18, 24, 28, 32 and 36 weeks. Fetal parameters, including biparietal diameter, femur length and abdominal circumference, were recorded. The daily growth rates for biparietal diameter, femur length, and fetal abdominal area were calculated and compared with those in a low-risk (non-diabetic) population. The growth rates in fetuses of women with satisfactory diabetic control (HbA1c < 6.5%) and unsatisfactory control (HbA1c > or = 6.5%) in the three trimesters were compared. RESULTS: A total of 174 diabetic pregnancies were included and a total of 997 ultrasound scans were performed. The growth rates for fetuses of mothers with diabetes mellitus were significantly higher than for those in the low-risk population. The z-scores for biparietal diameter, femur length, and fetal abdominal area were 0.18, 0.59 and 1.44, respectively. Fetuses of diabetic mothers with high HbA1c in the first trimester had significantly greater fetal abdominal area growth rate than those with normal HbA1c (fetal abdominal area z score of 1.7 vs. 0.75, P = 0.009). Although the fetal abdominal area z-scores in fetuses of diabetic mothers with high HbA1c in the second or third trimesters were also higher than those with normal HbA1c levels, the differences did not reach statistical significance. Maternal obesity did not influence the fetal growth rate. CONCLUSION: The rate of growth of fetuses of diabetic mothers differs from that of the normal population. Growth acceleration persists until the late third trimester. Moreover, periconceptional glucose control appears to have a significant effect on accelerated growth of the fetal abdominal area. PMID- 17083145 TI - Is self-efficacy a predictor of short-term post-surgical adjustment among Chinese women with breast cancer? AB - BACKGROUND: High self-efficacy (SE) is regarded as beneficial for cancer patients in facilitating adaptation and therefore desirable. However, this may not always be the case. DESIGN: A longitudinal cohort study of women receiving breast cancer surgery. Path analysis examined impact of high and low baseline SE scores on outcome. Post hoc analysis stratified outcome expectations by SE. METHODS: 405/529 eligible Chinese women aged 28-79 years receiving breast cancer surgery in six regional Hong Kong hospitals were interviewed within 1 week of surgery. After assessing SE, incongruence between expectancy and outcome of surgery (E OI), and psychological morbidity, 91% of women were followed for 1 month when psychological and social morbidity were assessed (follow-up). RESULTS: After adjustment for demographic and histopathological factors, psychological morbidity was predicted by E-OI. Women with high E-OI had more impairment of sexuality and self-image. Women with high SE had better self-image and relationships with friends, but tended to underestimate the negative consequences of surgery on appearance. This increased E-OI and thereby psychological morbidity. CONCLUSIONS: High post-surgical SE benefits early social adaptation, but also leads to under estimating the negative impacts of surgery, impairing psychological adjustment. High SE can thereby contribute indirectly and significantly to increased psychological morbidity. PMID- 17083146 TI - Osteoclast nuclei of myeloma patients show chromosome translocations specific for the myeloma cell clone: a new type of cancer-host partnership? AB - A major clinical manifestation of bone cancers is bone destruction. It is widely accepted that this destruction is not caused by the malignant cells themselves, but by osteoclasts, multinucleated cells of monocytic origin that are considered to be the only cells able to degrade bone. The present study demonstrates that bone-resorbing osteoclasts from myeloma patients contain nuclei with translocated chromosomes of myeloma B-cell clone origin, in addition to nuclei without these translocations, by using combined FISH and immunohistochemistry on bone sections. These nuclei of malignant origin are transcriptionally active and appear fully integrated amongst the other nuclei. The contribution of malignant nuclei to the osteoclast population analysed in this study was greater than 30%. Osteoclast myeloma clone hybrids contained more nuclei than normal osteoclasts and their occurrence correlated with the proximity of myeloma cells. Similar hybrid cells were generated in myeloma cell-osteoclast co-cultures, as revealed by tracing myeloma nuclei using translocations, bromo-deoxyuridine, or the Y chromosome of male myeloma cells in female osteoclasts. These observations indicate that hybrid cells can originate through fusion between myeloma cells and osteoclasts. In conclusion, malignant cells contribute significantly to the formation of bone resorbing osteoclasts in multiple myeloma. Osteoclast-myeloma clone hybrids reflect a previously unrecognized mechanism of bone destruction in which malignant cells participate directly. The possibility that malignant cells corrupt host cells by the transfer of malignant DNA may have been underestimated to date in cancer research. PMID- 17083147 TI - Case 24-2006: a woman with hypotension after an overdose of amlodipine. PMID- 17083142 TI - Human body fluid proteome analysis. AB - The focus of this article is to review the recent advances in proteome analysis of human body fluids, including plasma/serum, urine, cerebrospinal fluid, saliva, bronchoalveolar lavage fluid, synovial fluid, nipple aspirate fluid, tear fluid, and amniotic fluid, as well as its applications to human disease biomarker discovery. We aim to summarize the proteomics technologies currently used for global identification and quantification of body fluid proteins, and elaborate the putative biomarkers discovered for a variety of human diseases through human body fluid proteome (HBFP) analysis. Some critical concerns and perspectives in this emerging field are also discussed. With the advances made in proteomics technologies, the impact of HBFP analysis in the search for clinically relevant disease biomarkers would be realized in the future. PMID- 17083148 TI - Case 24-2006: a woman with hypotension after an overdose of amlodipine. PMID- 17083149 TI - Climate-change battle heats up. PMID- 17083150 TI - Hopes pinned on biofuel. PMID- 17083151 TI - Boost for Europe's research plans. PMID- 17083154 TI - Peril in the pink. PMID- 17083153 TI - Q & A. PMID- 17083155 TI - Abbott expands low cost Kaletra. PMID- 17083156 TI - Push to educate Mexican immigrants. PMID- 17083157 TI - [Tumor necrosis factor-alpha system in patients with gestational diabetes]. AB - Tumor necrosis factor-alpha (TNF-alpha) system is potentially involved in the development of insulin resistance during pregnancy. Plasma concentrations of TNF alpha and its soluble receptors sTNFR-1 and sTNFR-2 were measured in 80 patients with gestational diabetes (GDM) (mean age 29.0 +/- 4.9 years) and 30 pregnant women with normal glucose tolerance (NGT) (mean age 28.2 +/- 6.0 years). We found that patients with GDM had significantly higher levels of TNF-alpha in comparison to NGT women (1.71+/- 0.92 vs. 1.27 +/- 0.42 pg/ml, p = 0.0175). The differences remained statistically significant after adjusting for BMI (p = 0.027). Plasma levels of sTNFR-1 and sTNFR-2 were only slightly higher in patients with GDM (2.83 +/- 0.79 ng/ml vs. 2.55 +/- 0.99 ng/ ml, p = 0.057 and 7.46 +/- 2.21 ng/ml vs. 6.83 +/- 1.46 ng/ml, p=0.206, respectively). In the group with GDM TNF-alpha concentrations correlated with sTNFR-1 (r = 0.444, p = 0.00008), sTNFR-2 (r = 0.364, p = 0.0016) and with C-peptide concentrations (r = 0.318, p = 0.016), whereas in women with NGT - only with triglyceride levels (r = 0.50, p = 0.024). Multivariate linear regression analysis revealed that early pregnancy BMI was the most predictive indicator of TNF-alpha concentrations in GDM women (p=0.008). In NTG group triglyceride concentrations, as well as BMI in early pregnancy and at the time of sampling were significant predictors, explaining together 62% of the variance in TNF-alpha concentration. In conclusion, increased TNF-alpha concentrations in women with GDM class G1 indicates its contribution to the development of insulin resistance during pregnancy, but the lack of the differences in sTNFR concentrations between the groups studied suggests only moderate TNF-alpha system activation in relatively slim patients treated with diet. PMID- 17083158 TI - [Comparative analysis of CD56 lymphocytes and CD69 antigen expression in endometrium during decidualization]. AB - INTRODUCTION: The exceptionality of endometrium, differentiating it from other epithelial tissues is related with its ability to regulate the number and activity of infiltrating immunological cells according to menstrual cycle phases. The aim of this study was a comparative evaluation of the number of CD56 positive cells, as well the analysis of CD69 antigen expression during decidualization. MATERIALS AND METHODS: The expression of CD56 and CD69 antigens was analyzed using the immunohistochemistry method in 20 decidual tissue samples derived after curettage following spontaneous abortion and in 38 endometrial tissue samples obtained during hysterectomy performed because of benign myoma diagnosis. The endometrial tissue samples were divided regarding to the menstrual cycle phases. Following phases were distinguished: early proliferative, mid-proliferative, late proliferative, early secretory, mid secretory and late secretory. RESULTS: The higher CD56 positive cells were recognized in the endometrium during mid secretory cycle phase. The number of CD56 positive cells was significantly lower in endometrium, independent of menstrual cycle phase in comparison to endometrium following abortion. The expression of CD69 antigen was observed to increase beginning with early secretory cycle phase and remained at high level during the secretory cycle phase and the abortion. CONCLUSIONS: The observed differences in the number of CD56 positive cells during decidualization without the changes in immune cells activity evaluated using CD69 antigen expression may indicate the participation of activated mononuclear cells in the normal development of pregnancy beginning with implantation. PMID- 17083159 TI - [Vascular adhesion protein-1 (VAP-1) in inflammatory process]. AB - VAP-1 is an adhesion molecule expressed on the surface of endothelial cells. It plays an important role in the adhesion and migration of leukocytes to the sites of inflammation. The purpose of this article is to present current knowledge of structure, biological function of VAP-1 and its use in clinical practice. PMID- 17083160 TI - [Health properties of shark oil]. AB - Fish oils are the source of nutrients important for health maintenance. The most significant are essential fatty acids (EFA) of n-3 family, alkylglycerols and squalene. N-3 EFA are of great importance in atherosclerosis prevention. Alkylglycerols and squalene are modulators of immunity to infections and cancer. Shark liver oil contains great amounts of alkylglycerols and squalene, and moderate of n-3 EFA. Therefore, it is used as an adjunctive agent in cancer therapy, especially in radiotherapy, and in the treatment of infectious diseases. PMID- 17083161 TI - Current clinical uses of intravenous immunoglobulin. PMID- 17083162 TI - Assisted dying. PMID- 17083163 TI - p38 mitogen-activated protein kinase: a novel modulator of hyperpolarization activated cyclic nucleotide-gated channels and neuronal excitability. PMID- 17083164 TI - Membrane estrogen receptors and energy homeostasis. PMID- 17083165 TI - Summary and recommendations of the 12th informal consultation of the WHO Global Polio Laboratory Network, June 2006. PMID- 17083166 TI - Health conditions for travellers to Saudi Arabia for the pilgrimage to Mecca (Hajj). PMID- 17083167 TI - Amino-functionalized SBA-15 type mesoporous silica having nanostructured hexagonal platelet morphology. AB - Amino-functionalized SBA-15 type mesoporous silicas having unique hexagonal platelet morphologies with short channels (100-300 nm) running parallel to the thickness of the nanostructured hexagonal platelet type morphologies have been directly synthesized by co-condensation of aminopropyltriethoxysilane (APTES) and sodium metasilicate as a silica source in the presence of Pluronic P123 triblock copolymer as a structure directing agent. PMID- 17083168 TI - Cognitive functioning of adolescents and young adults with meningomyelocele and level of everyday physical activity. AB - PURPOSE: To explore whether cognitive functioning in patients with meningomyelocele (MMC) is related to level of everyday physical activity. METHOD: In a cross-sectional study in 14 patients with MMC (aged 14 - 26 years) a neuropsychological test battery was administered to each patient. Everyday physical activity of the patients was measured with an accelerometry-based Activity Monitor (AM), and compared to 14 healthy comparison subjects. RESULTS: Intellectual ability of 10 patients was in the normal range. For memory and verbal learning, executive functioning, divided attention and reaction speed subnormal scores were present in six or more patients. Time spent on dynamic activities was low in patients with MMC as compared to healthy subjects. After controlling for intellectual ability, we found that in patients with MMC (i) executive functioning was positively related, and (ii) word production was negatively related to everyday physical activity. CONCLUSION: We found some indication that specific impairments in executive functioning might be related to everyday physical activity of adolescents and young adults with MMC. PMID- 17083169 TI - Exertional symptoms and exercise capacity in individuals with brain injury. AB - PURPOSE: To examine factors affecting cycling exercise performance in individuals with acquired brain injury. METHODS: Thirty individuals with acquired brain injury and 18 sedentary controls (SC) participated. Heart rate, bicycle power output and rating of perceived exertional (RPE) were recorded, throughout incremental cycle ergometer exercise. The SC group and 18 moderately impaired individuals from the ABI group performed a 25-W (B25) protocol. The remaining 12 individuals performed a 10-W protocol (B10). RESULTS: The B10 group terminated exercise at the lowest RPE, percentage age predicted maximal heart rate (% APMHR) and bicycle power output, followed by the B25 and then the SC group (RPE: Kruskal - Wallis test P < 0.001, %APMHR and bicycle power output: one-way ANOVA P < 0.01). RPE was correlated with %APMHR and percentage of peak bicycle output (B10 group: R2 0.1 to 0.67; B25 group: 0.69 - 0.83; SC group: 0.76 - 0.91). There was no difference in RPE at the same relative work intensity between the B25 and the sedentary control group (P > 0.05). Forward regression analysis revealed fatigue levels were predictive of %APMHR at test termination (beta = -0.411, P < 0.05) and quadriceps strength was predictive of peak bicycle power output (beta = 0.612, P < 0.05). CONCLUSIONS: Individuals with brain injury terminated exercise at lower exercise intensities but rated exertion no differently from healthy individuals. General fatigue levels predicted %APMHR and quadriceps strength predicted peak bicycle power output. PMID- 17083170 TI - Self-worth, perceived competence, and behaviour problems in children with cerebral palsy. AB - PURPOSE: To examine the relevance of physical disabilities for self-worth and perceived competence in children with cerebral palsy (CP), and to examine associations between behaviour problems and self-worth and perceived competence. METHODS: The Harter scales for self-worth and perceived competence and a new scale for perceived motor competence were used in a sample of 80 children with CP. Their motor functioning was assessed with the Gross Motor Functioning Measure (GMFM) and behaviour problems with the Child Behaviour Check List administered to parents. RESULTS: Self-worth and perceived competence for children with CP were comparable to the Dutch norm sample, except for perceived athletic competence. Within the CP sample, the GMFM showed a domain-specific effect on perceived motor competence. In the multivariate analysis, internalizing problems were associated negatively with all perceived competence scales and self-worth, whereas aggression was positively associated with perceived motor competence, physical appearance, and self-worth. CONCLUSIONS: Children with CP appear resilient against challenges posed to their self-worth caused by their disabilities. The relevance of the physical disability appears to be domain-specific. For internalizing problems and aggression, different theoretical models are needed to account for their associations with self-worth and perceived competence. PMID- 17083171 TI - Syringe driver guidelines. PMID- 17083172 TI - Determinants of shuttle run performance in the prediction of peak VO2 in wheelchair users. AB - PURPOSE: The purpose of this study was to determine the impact of ergonomic and environmental variations on indoor shuttle run (SR) performance in wheelchair sportsmen. METHODS: Eleven experienced male wheelchair sportsmen performed three 25-m SRs in random order with varying turning capacity (TC) and mechanical resistance (MR): condition NN where participants used their sports wheelchair on a tartan surface, condition RN with increased MR, and condition RD with limited TC. Metabolic data were continuously recorded using a portable K4b(2) system. RESULTS: Friedman ANOVA with Wilcoxon a posteriori testing indicated similar VO(2peak) values in all three tests. SR performance, however, was significantly different across the three test conditions (NN: 536.18 +/-119.09 s; RN: 488.82+/ 119.84 s; RD: 404.91+/-88.41 s). SR performance contributed for 28% of the explained variance of the measured VO(2peak). The addition of TC or MR or both increased the explained variance to 32, 38 and 41%, respectively. CONCLUSIONS: These findings demonstrate a significant impact of variations in floor surface and wheelchair-user interface on SR performance. The findings also suggest that strong reservations have to be made regarding the validity of a SR test, as predictor of VO(2peak) in a wheelchair user population. PMID- 17083173 TI - Communication and eating proficiency in 125 females with Rett syndrome: The Swedish Rett Center Survey. AB - PURPOSE: To describe communication and eating proficiency and related factors in Swedish females with Rett syndrome, from a parental/caregiver view. METHOD: Data from a questionnaire were used and approached descriptively, qualitatively and comparatively. Parents/caregivers of 125 females with Rett syndrome completed the survey. The mean age of the females was 19.6 years. RESULTS: Communication: expressive language was limited and the most common way of expressing will was by producing motor acts, 50% of which were higher level of language (eye- and finger pointing and gestures). Results also showed communicative behaviour was demonstrated during social activities. Eating proficiency: For eight specific types of eating difficulties, the majority of the females suffered from none, one or two of them. There were significant associations of some specific eating difficulties with breathing dysfunctions and food consistency, but not with dental or oral problems. CONCLUSIONS: Results from this study have confirmed that verbal communication in Rett syndrome is limited. Despite a lack of verbal communication the females in this study were reported to need and enjoy social interplay. Outcomes concerning eating were generally good. For optimal intervention, it is recommended that family and professionals work together to evaluate eating situations and communication ability for females with Rett syndrome. PMID- 17083174 TI - A burning desire to understand genetic mutations in gastroenterology. PMID- 17083175 TI - Assessment of symptom severity and functional status in patients with carpal tunnel syndrome: reliability and functionality of the Turkish version of the Boston Questionnaire. AB - OBJECTIVES: The aim of this study was to develop a Turkish version of the Boston Questionnaire and assess its reliability and validity. METHODS: Sixty-seven patients with idiopathic carpal tunnel syndrome were included in the study. The Turkish version of Boston Questionnaire was obtained after translation process, and was then administered to subjects twice within seven days. Reliability was assessed by internal consistency (Cronbach's alpha and item-total correlation), and reproducibility. Validity was examined by correlating the Boston Questionnaire scores to general health status (Short Form-36), pain severity (Visual Analogue Scale) and pinch and grip strength measures. RESULTS: Reliability of the Turkish version was very good, with high internal consistency (Cronbach's alpha 0.82 for symptom severity scale, and 0.88 for functional status scale), and reproducibility (Pearson correlation coefficient 0.60 for symptom severity scale, and 0.77 for functional status scale). The Boston Questionnaire scores were correlated with Visual Analogue Scale, physical functioning, physical role, bodily pain and emotional role subscales of Short Form-36, pinch and grip strength scores to obtain coefficients for external construct validity. CONCLUSION: Adaptation of the Boston Questionnaire for use in Turkey was successful. Our results seem to support previous finding of the English version, indicating that it is valid and reliable. PMID- 17083177 TI - Artifacts caused by contrast agents and patient movement in cardiac PET-CT. PMID- 17083176 TI - Reliability of the Chinese version of the Activities-specific Balance Confidence Scale. AB - PURPOSE: To translate the Activities-specific Balance Confidence (ABC) Scale into a Chinese version and assess the reliability between Chinese versions and between Chinese and English versions of this outcome measure. METHOD: Descriptive study using a 4-week test-retest design. Data were collected from a convenience sample of community living Chinese immigrants. Of the 79 participants, data from 71 subjects were included in the analysis. Two subsamples were formed to assess the reliability between Chinese versions (n = 33) and between Chinese and English versions (n = 38) of the scale. RESULTS: Internal consistency of the ABC-C was 0.98. Test-retest reliability was intraclass correlation coefficient (ICC) = 0.87 (95% CI, 0.76 - 0.93) for the Chinese versions and ICC = 0.88 (95% CI, 0.78 - 0.94) for Chinese and English versions. The total group ICC = 0.90 (95% CI, 0.84 0.94). CONCLUSIONS: Balance confidence has been identified as an important area for clinical and research inquiry however collecting this information from Chinese speaking individuals has been limited by a lack of language specific measures. The Chinese version of the ABC has demonstrated acceptable measurement properties in this sample and should permit measurement of this unique construct in the Chinese population. PMID- 17083178 TI - Alphabetical prejudice in team discussions (or would Zebedee ever get seen on a ward round). AB - PURPOSE: To ascertain whether the order in which patients are discussed in a team meeting determines the time spent on discussion. DESIGN: Prospective study over 18 consecutive multidisciplinary team meetings. SETTING/SUBJECTS: Multidisciplinary team meetings on a Brain Injury Rehabilitation Unit, Edinburgh. MAIN OUTCOME MEASURE: Time spent discussing each case. RESULTS: Patients discussed early on received 3-4 minutes more discussion time than those later on. This was highly significant on a one-way analysis of variance (P < 0.001). CONCLUSIONS: Preferential treatment of patients who come at the start of the team meeting is a real phenomenon. Such alphabetical prejudice, where it exists, should be addressed. PMID- 17083179 TI - Prognosis and prognostic factors in ADL-dependent stroke patients during their first in-patient rehabilitation--a prospective multicentre study. AB - PURPOSE: This prospective longitudinal study aimed at a description of ADL functioning and at an analysis, which clinical instruments predict independency 6 months after discharge from in-patient rehabilitation following disabling first stroke. METHOD: A total of 147 patients with disabling first stroke were recruited from three neurological rehabilitation centres. They were assessed during their first in-patient rehabilitation with the NIH Stroke Scale, Hemispheric Stroke Scale (HSS), Activity Index, Barthel Index, Neurobehavioural Rating Scale, Cornell Depression Scale, and the prognostic prediction of the rehabilitation neurologist. The 6 months outcome was established with the Nottingham Extended ADL Scale (NEADL) and by the legal dependency status. RESULTS: The best prediction of 6 months outcome as measured by the NEADL was obtained by the HSS and a multivariate model that also included the physician's prognostic prediction, both early in rehabilitation and at discharge. The dependency status was best predicted by the HSS and patient's sex. CONCLUSION: Clinical scales performed during rehabilitation such as the HSS are valid for prediction of ADL-dependency 6 months after discharge. PMID- 17083180 TI - The impact of psychosocial factors on neck pain and disability outcomes among primary care patients: results from the UCLA Neck Pain Study. AB - PURPOSE: The purpose of this study is to estimate the associations of psychosocial factors with pain and disability outcomes among neck-pain patients enrolled in a randomized clinical trial of chiropractic treatments. METHODS: Neck pain patients were randomized to one of 8 modes of chiropractic treatment. Health status and psychosocial variables were measured at baseline. Changes in neck pain severity and disability from baseline to 6 months were the primary outcome variables. Multivariable regression models were used to estimate effects of psychosocial variables adjusted for potential confounders. RESULTS: Of 960 eligible patients, 336 were enrolled and 80% were followed up through 6 months. Coping strategies involving self-assurance resulted in better disability outcomes, whereas getting angry or frustrated resulted in worse pain and disability outcomes. Participants with high levels of social support from individuals were more likely to experience clinically meaningful reductions in pain and disability. No consistent relations of internal health locus of control, and physical and psychological job demands with improvements in pain and disability were detected. CONCLUSIONS: We found some evidence that certain coping strategies and types of social support are associated with pain and disability outcomes in this population of largely subacute and chronic neck-pain patients. PMID- 17083181 TI - Impact of visual impairment on service and device use by individuals with age related macular degeneration (AMD). AB - PURPOSE: To assess the patient-reported use of services, supplements, and devices among individuals with age-related macular degeneration (AMD) and evaluate the impact of visual impairment level on this use. METHOD: Data for this study were collected using two instruments, the AMD Health and Impact Questionnaire and the Daily Living Tasks Dependent on Vision questionnaire (DLTV). Both questionnaires were mailed to members of the Macular Degeneration Partnership. The study was approved by an IRB and respondents provided consent before participating. Respondents' visual acuity (VA) was estimated using scores from the DLTV, while use of services and devices was collected from the AMD Questionnaire. De identified data were analysed in SAS. RESULTS: Of 803 respondents, 56% were male and the mean age was 73 years. Use of services (e.g., counseling, rehabilitation), and devices significantly increased as VA decreased. Using standard US costs, costs for services, supplements, and devices ranged from 506 1619 US dollars depending on VA. CONCLUSION: There are substantial differences in service and device use with increased AMD severity. Delaying progression of AMD could result in considerable cost savings. PMID- 17083182 TI - The nature and predictors of stroke knowledge amongst at risk elderly persons in Brisbane, Australia. AB - PURPOSE: The aim of the study was to investigate the nature, extent, and predictors of stroke knowledge amongst people at risk of stroke. METHOD: A total of 273 questionnaires were distributed to three types of sites in metropolitan Brisbane (retirement villages, senior citizens and bowls clubs), and this strategy yielded a response rate of 37%. Of the surveys returned, 75 were retained in this study on the basis that: (a) participants met our criteria for being at risk of stroke (i.e., they reported one or more modifiable stroke risk factor, such as hypertension), and (b) they returned a completed Stroke Knowledge Test (SKT). RESULTS: Descriptive analyses revealed the overall level of stroke knowledge in this group was fair (approximately 50% of SKT items answered correctly, on average). SKT performance trends showed that participants did not have misconceptions about stroke but that more than 50% of the sample did not know the answer to seven out of 20 SKT items. Some overlap was identified regarding the issues about which participants in this and other previously researched groups admit they lack knowledge, such as the extent of increased stroke risk associated with smoking; however, the number of knowledge gaps identified in this sample was almost double that reported previously amongst stroke survivors and the general community. Analyses undertaken to explore determinants of stroke knowledge revealed age and education but not risk factor variables as significant predictors. CONCLUSION: Overall, findings suggest that it is presently difficult to predict an individual's understanding of stroke and that there is a need to increase stroke education, especially if this can address issues that people at risk of stroke admit they do not understand. PMID- 17083183 TI - Service delivery trends for a physical rehabilitation outreach program. AB - PURPOSE: Evaluate clinical service characteristics of a physical rehabilitation outreach clinic. METHOD: A retrospective analysis on service-related factors and patient information (every clinical encounter between 1983 and 2002) was performed on the Terry Fox Mobile Clinic. The Terry Fox Mobile Clinic provided multidisciplinary physical rehabilitation outreach services for adults with disabilities in Eastern and North Eastern Ontario, Canada (on-site visits by rehabilitation specialists). Subjects include 4816 clients with physical disabilities from Eastern and North Eastern Ontario, Canada. Patient data (age, residence, diagnosis, inpatient/outpatient), clinic data (type, location, totals), team data (clinical disciplines), and assistive device utilization over the last 15 years of data were used for analysis. RESULTS: Over 15 years, 4816 clients (1032 clinic days, 1.46 patient-contact ratio) were seen by the outreach team. Single discipline days were the best 'number of clinic days' predictor. Following 1999, the 35-64 age group became predominate (instead of over-65). Most patients lived in their own residence. Stroke, amputations, neck and back pain, and cerebral palsy accounted for 53% of patient diagnoses. Wheelchairs and orthoses were the highest ranked assistive device consultations/prescriptions (total = 6304). Physicians were involved with the majority of patient contacts. Most clinic activity occurred at sites within 100 km driving distance. Sites over 200 km away had the lowest correlation with the number of clinic days. CONCLUSIONS: The 15 years of physical rehabilitation outreach service data is valuable for planning new outreach programs, benchmarking existing services, and telerehabilitation comparisons. Multidisciplinary physical rehabilitation can make a positive contribution to healthcare that extends beyond direct patient contact time. PMID- 17083184 TI - Comments on van Brakel et al.'s Participation Scale. PMID- 17083185 TI - Doe v. Roe. PMID- 17083186 TI - Health Insurance Association of America v. Corcoran. PMID- 17083187 TI - Abram v. Children's Hospital of Buffalo. PMID- 17083188 TI - Elbaum v. Grace Plaza of Great Neck. PMID- 17083189 TI - In re Klein. PMID- 17083190 TI - Fosmire v. Nicoleau. PMID- 17083191 TI - New York State Society of Surgeons v. Axelrod. PMID- 17083192 TI - Anderson v. Strong Memorial Hospital. PMID- 17083193 TI - Westchester County Medical Center v. Hall. PMID- 17083194 TI - Health Insurance Association of America v. Corcoran. PMID- 17083195 TI - People v. Thomas. PMID- 17083196 TI - In re Salisbury. PMID- 17083197 TI - Human recombinant PLD2 can repress p65 activity of guinea pigs of chronic asthma in vivo. AB - This article is to investigate the effect of human recombinant phospholipase D2 (rhPLD2) in vivo on the expression of nuclear transcription factor p65 in chronic asthma of guinea pigs. After treating the guinea pigs with chronic asthma by rhPLD2, the crude nuclear extraction was assayed with TransAM Transcription Factor Assay Kit for the activity of pulmo tissue nuclear transcription factor p65. Compared with the healthy guinea pigs, the activity of nuclear transcription factor p65 in guinea pigs of chronic asthma is much higher than that of control groups. Our results showed that rhPLD2 markedly depressed the activity of p65 when the guinea pigs were attacked by chronic asthma. PMID- 17083198 TI - Expression, purification and identification of recombinant mouse interleukin 21 protein in E. coli. AB - Interleukin 21 (IL-21) is a novel type I cytokine that is significantly homologous to IL-2, IL-4 and IL-15. Its receptor complex contains gammac chain which is also a component of receptors for IL-2, IL-4, IL-7, IL-9 and IL-15, so there may be overlapping or relevancies in their biological functions. IL-21 is capable of co-stimulating mature T cells, B cells, NK cells, and of stimulating CD16 expression on the surface of NK cells to induce ADCC in innate immune response. It can also strengthen the anti-tumor effect of the cellular immunity, especially via enhancing the activities of NK and antigen specific CTL cells. Thus, IL-21 is a potential useful therapeutic molecule for immunotherapy of malignancies, by eliciting innate and adaptive anti-tumor responses in tumor bearing hosts. In order to study the biological functions of IL-21, we constructed a mIL-21 prokaryotic expression plasmid and expressed the recombinant mIL-21 protein in E. coli in present study. The recombinant plasmid pET28a/mIL-21 with a carboxyl terminal His-tag was subcloned from the pcDNA3.1/mIL-21 and expressed in E. coli. The induced protein was detected by SDS-PAGE, and identified by Western-blot assay with anti-mIL-21 antibody. The recombinant protein was purified via Ni+ affinity chromatography, and renatured with GSH/GSSG system. Our mouse T cell proliferation experiment showed that the recombinant mIL 21 protein could enhance the mouse T cell proliferation either by itself alone or in the presence of Con A. PMID- 17083199 TI - In re Beth Israel Medical Center. PMID- 17083200 TI - Ughetto v. Acrish. PMID- 17083201 TI - Delio v. Westchester County Medical Center. PMID- 17083202 TI - Correction of thiazide-induced hypokalemia prevents hyperglycemia and incident diabetes mellitus. PMID- 17083203 TI - Building a scientific base for nutrition care of Hispanic nursing home residents. AB - By 2030, one-quarter of the U.S. Hispanic population will be aged 80 or over. At least 4.5 million of them will require long-term care, most probably in nursing homes. Although Hispanic families traditionally care for elders at home despite their declining cognitive and physical function, family ability to provide such care is eroding due to acculturation, increasing the likelihood of nursing home admission. A synthesis of two studies (findings from a focus group and a Hispanic data subset from a federally funded grant) found that Hispanic nursing home resident food intake could be improved by traditional entrees, appealing presentations, and flour tortillas from a supplier recommended by Mexican American families. There were no statistically significant differences between Anglo and Hispanic residents on study variables, but results suggest that there may be clinically meaningful differences among such residents on Mini Mental State Examination scores, prealbumin results, the Geriatric Depression Scale, and the Katz Activities of Daily Living that call for further investigation. PMID- 17083204 TI - NaCl-type frameworks of [M(pyrazine)2NO2]ClO4 (M = Co, Cu), the first examples containing mu(1,3)-nitrito bridges showing antiferromagnetism. AB - Two new 3D NaCl-type frameworks of [M(pyrazine)2NO2]ClO4 (M = Co, Cu), consist of (4, 4) sheets of metal-pyrazine connected by mu(1,3)-nitrito, representing the first examples of mu(1,3)-nitrito bridges and exhibiting antiferromagnetism. PMID- 17083205 TI - New barium cobaltite series Ba(n+1)Co(n)O(3n+3)(Co8O8): intergrowth structure containing perovskite and CdI2-type layers. AB - Single crystals of two new cobaltites, Ba2Co9O14 and Ba3Co10O17, were obtained from the flux of K2CO3 in the temperature range 800-890 degrees C. They crystallize in an intergrowth structure containing perovskite block and CdI2-type layers and can be attributed to the n = 1 and 2 members in a new intergrowth series of cobaltites, Ba(n+1)Co(n)O(3n+3)(Co8O8). Both Ba2Co9O14 and Ba3Co10O17 are metastable and transform into the known 2H-perovskite-related oxides at high temperature. PMID- 17083206 TI - Novel layered copper phosphoramidate, which contains six-membered rings made of five different elements. AB - Copper(II) perchlorate and (PhCONH)P(O)(OH)2 (H2bpa) react in methanol to yield [Cu(Hbpa)2]n, a novel layered solid that displays several interesting features. Unlike the previously reported copper phosphonates, a rare hexacoordination around the metal is observed in the title compound because of the amide C=O coordination to Cu in addition to the phosphoramidate P-O coordination. The six membered chelate rings present in the title compound are made up of five different elements (Cu, P, C, N, and O). PMID- 17083207 TI - A novel cascade supramolecular complex with a reversible nanosized 18-component H bonded {(C6H6)6(SbF6)12}12- cage unit templated by a chiral metal-organic complex. AB - A novel cascade supramolecular complex [Ag(L)](SbF6).C6H6 with a reversible nanosized {(C6H6)6(SbF6)12}12- H-bonded cage unit was reported. PMID- 17083208 TI - Metal-metal bonding in sterically frustrated dipalladium species. AB - Stoichiometric-deficient lithiation of (2,6-diisopropylphenyl)(4-methylpyridin-2 yl)amine and reaction with [(cod)PdCl2] (cod = 1,5-cyclooctadiene) yield a dimeric Pd complex. X-ray structural analysis of this complex reveals a very short Pd-Pd distance (2.429 A). Topological analysis of the electron density and the electron localization function from scalar relativistic density functional theory calculations clearly indicate a Pd(I)-Pd(I) sigma-bonding interaction, for which the corresponding occupied localized orbital can be identified. PMID- 17083209 TI - Two polyknotted topological isomers of copper(I) 3,5-bis(4-pyridyl)pyrazolates. AB - Two unprecedented 3D polyknotted isomers, arisen from different linkage modes of SCN-, were obtained from 3,5-bis(4-pyridyl)-1H-pyrazole (Hbppz) and CuSCN under different conditions. PMID- 17083210 TI - Expanding the scope of sulfur-centered Arbuzov rearrangement in diethyl/di-n propyl sulfite for the synthesis of mixed-ligand di-n-butyltin alkanesulfonates. AB - A one-pot reaction between di-n-butyltin oxide and diethyl/di-n-propyl sulfite in the presence of an equimolar amount of alkyl iodide proceeds via sulfur-centered Arbuzov rearrangement to afford the corresponding di-n-butyltin (alkoxy)alkanesulfonates n-Bu2Sn(OR')OS(O)2R [R = R' = Et (1), n-Pr (2); R = Me, R' = Et (3), n-Pr (4)]. The compounds 1 and 3 react with methylphosphonic acid under mild conditions to give [n-Bu2Sn(OS(O)2R)OP(O)(OH)Me]n [R = Et (5), Me (6), respectively]. PMID- 17083211 TI - Bidentate phosphine ligand based Fe2S2-containing macromolecules: synthesis, characterization, and catalytic electrochemical hydrogen production. AB - The reaction of [Fe2(CO)6(mu-SCH2)2NCH2CH2N(mu-SCH2)2Fe2(CO)6] (1) with 1,2 bis(diphenylphosphino)ethane in the presence of Me3NO.2H2O affords two structurally different metallomacromolecules: a dimer of the type [{Fe2(CO)5(mu SCH2)2NCH2CH2N(mu-SCH2)2Fe2(CO)5}(Ph2PCH2)2] (2) and a tetramer species containing eight iron centers with an overall formula of [{Fe2(CO)6(mu SCH2)2NCH2CH2N(mu-SCH2)2Fe2(CO)5}2(Ph2PCH2)2] (3). Their structures have been determined by X-ray crystallography, showing one macrocyclic complex (2) and one linear complex (3). Electrochemical hydrogen evolution catalyzed by these two complexes with ca. 80-90 single-run turnovers is observed, indicating good potential as catalysts for future applications. PMID- 17083212 TI - NMR observations of 13C-enriched coenzyme B12 bound to the ribonucleotide reductase from Lactobacillus leichmannii. AB - The 13C NMR resonance and one-bond 1H-13C coupling constants of coenzyme B12 enriched in 13C in the cobalt-bound carbon have been observed in the complex of the coenzyme with the B12-dependent ribonucleotide reductase from Lactobacillus leichmannii. Neither the 13C NMR chemical shift nor the 1H-13C coupling constants are significantly altered by binding of the coenzyme to the enzyme. The results suggest that ground-state Co-C bond distortion is not utilized by this enzyme to activate coenzyme B12 for C-Co bond homolysis. PMID- 17083213 TI - Symmetric and unsymmetric "dumbbells" of Ru2-alkynyl units via C-C bond formation reactions. AB - Oxidative homocoupling (Glaser) reaction of Ru2 compounds bearing peripheral ethyne resulted in symmetric dimers. Cross-coupling (Sonogashira) reaction between Ru2 compounds bearing peripheral iodo and ethyne groups yielded an unsymmetric dimer. Voltammetric data indicated that Ru2 units in the symmetric dimers are noninteracting, and the unsymmetric dimer is best described as a weakly coupled push-pull compound. PMID- 17083214 TI - Phototriggered S --> O isomerization of a ruthenium-bound chelating sulfoxide. AB - We have prepared and characterized [Ru(bpy)2(OS)]+ (bpy = 2,2'-bipyridine; OS = 2 methylthiobenzoate) and the chelating sulfoxide S-[Ru(bpy)2(OSO)]+ (OSO = methylsulfonylbenzoate) by 1H NMR (1-D and 2-D COSY), IR, UV-visible spectroscopy, electrochemistry, and X-ray crystallography. Many of the metrical and crystal parameters are nearly identical between the two structures. The sulfoxide is produced from m-cpba oxidation of the thioether. Photolysis of S [Ru(bpy)2(OSO)]+ results in a dramatic shift in the metal-to-ligand charge transfer (3MLCT) transition from 396 to 496 nm, with a new higher-energy 3MLCT transition appearing at 355 nm. Concomitant with this change, the Ru3+/2+ reduction potential shifts from 1.25 V vs Ag/AgCl to 0.9 V vs Ag/AgCl. These changes are ascribed to phototriggered excited-state isomerization of the sulfoxide from S- to O-bonded. Examination of the 1H NMR spectra in a CD3OD solvent before, during, and after irradiation shows the presence of two O-bonded complexes that revert to the structurally characterized S-bonded ground state. This represents the first report of isomerization of a chelating sulfoxide in a photochromic Ru complex. PMID- 17083215 TI - Iron-only hydrogenase mimics. Thermodynamic aspects of the use of electrochemistry to evaluate catalytic efficiency for hydrogen generation. AB - Voltammetry is widely used for the evaluation of iron-only hydrogenase mimics and other potential catalysts for hydrogen generation using various dipolar aprotic solvents. Effective catalysts show enhanced current in the presence of a proton donor at the potential where the catalyst is reduced. To facilitate the comparison of catalytic efficiencies, this paper provides a simple means of calculating the standard potential for reduction of the acid, HA, according to the half reaction 2HA + 2e- <==> H2 + 2A-. This standard potential depends on the pKa of HA in the solvent being used. It is thermodynamically impossible for reduction of HA to occur at less negative potentials than the standard potential, and the most effective catalysts will operate at potentials as close as possible to the standard potential. In addition, direct reduction of HA at the electrode will compete with the catalyzed reduction, thus complicating evaluation of the rate of the catalyzed reaction. Glassy carbon electrodes, commonly used in such evaluations, show a quite large overpotential for direct reduction of HA so that the necessary corrections are small. However, catalysis at very negative potentials will be contaminated by significant direct reduction of HA at glassy carbon. It is demonstrated that direct reduction can be almost completely suppressed by using a mercury or amalgamated gold electrode, even at very negative potentials. PMID- 17083216 TI - Synthesis of molybdenum complexes that contain "hybrid" triamidoamine ligands, [(hexaisopropylterphenyl-NCH2CH2)2NCH2CH2N-aryl]3-, and studies relevant to catalytic reduction of dinitrogen. AB - In the Buchwald-Hartwig reaction between HIPTBr (HIPT = 3,5-(2,4,6-i Pr3C6H2)2C6H3 = hexaisopropylterphenyl) and (H2NCH2CH2)3N, it is possible to obtain a 65% isolated yield of (HIPTNHCH2CH2)2NCH2CH2NH2. A second coupling then can be carried out to yield a variety of "hybrid" ligands, (HIPTNHCH2CH2)2NCH2CH2NHAr, where Ar = 3,5-Me2C6H3, 3,5-(CF3)2C6H3, 3,5 (MeO)2C6H3, 3,5-Me2NC5H3, 3,5-Ph2NC5H3, 2,4,6-i-Pr3C6H2, or 2,4,6-Me3C6H2. The hybrid ligands may be attached to Mo to yield [hybrid]MoCl species. From the monochloride species, a variety of other species such as [hybrid]MoN, {[hybrid]MoN2}Na, and {[hybrid]Mo(NH3)}+ can be prepared. [Hybrid]MoN2 species were prepared through oxidation of {[hybrid]MoN2}Na species with ZnCl2, but they could not be isolated. [Hybrid]Mo=N-NH species could be observed as a consequence of the protonation of {[hybrid]MoN2}- species, but they too could not be isolated as a consequence of a facile decomposition to yield dihydrogen and [hybrid]MoN2 species. Attempts to reduce dinitrogen catalytically led to little or no ammonia being formed from dinitrogen. The fact that no ammonia was formed from dinitrogen in the case of Ar = 3,5-Me2C6H3, 3,5-(CF3)2C6H3, or 3,5-(MeO)2C6H3 could be attributed to a rapid decomposition of intermediate [hybrid]Mo=N-NH species in the catalytic reaction, a decomposition that was shown in separate studies to be accelerated dramatically by 2,6-lutidine, the conjugate base of the acid employed in the attempted catalytic reduction. X-ray structures of [(HIPTNHCH2CH2)2NCH2CH2N{3,5-(CF3)2C6H3}]MoCl and [(HIPTNHCH2CH2)2NCH2CH2N(3,5 Me2C6H3)]MoN2}Na(THF)2 are reported. PMID- 17083217 TI - Synthesis of [(HIPTNCH2CH2)3N]V compounds (HIPT = 3,5-(2,4,6-i-Pr3C6H2)2C6H3) and an evaluation of vanadium for the reduction of dinitrogen to ammonia. AB - Green [HIPTN3N]V(THF) ([HIPTN3N]3- = [(HIPTNCH2CH2)3N]3-, where HIPT = 3,5-(2,4,6 i-Pr3C6H2)2C6H3) can be prepared in a 70-80% yield via the addition of H3[HIPTN3N] to VCl3(THF)3 in THF, followed by the addition of LiN(SiMe3)2. From [HIPTN3N]V(THF), the following have been prepared: {[HIPTN3N]VN2}K, [HIPTN3N]V(NH3), [HIPTN3N]V=NH, [HIPTN3N]V=NSiMe3, [HIPTN3N]V=O, [HIPTN3N]V=S, and [HIPTN3N]V(CO). No ammonia is formed from dinitrogen using {[HIPTN3N]VN2}K, [HIPTN3N]V=NH, or [HIPTN3N]V(NH3) as the initial species under conditions that were successful in the analogous [HIPTN3N]Mo system. X-ray structural studies are reported for [HIPTN3N]V(THF) and [HIPTN3N]V(NH3). PMID- 17083218 TI - Fluoride abstraction and reversible photochemical reduction of cationic uranyl(VI) phosphine oxide complexes. AB - The syntheses, structural and spectroscopic characterization, fluoride abstraction reactions, and photochemical reactivity of cationic uranyl(VI) phosphine oxide complexes are described. [UO2(OPPh3)4][X]2 (1a, X = OTf; 1b, X = BF4) and [UO2(dppmo)2(OPPh3)][X]2 (2a, X = OTf; 2b, X = BF(4)) are prepared from the corresponding uranyl(VI) chloride precursor and 2 equiv each of AgX and phosphine oxide. The BF4- compounds 1b and 2b are prone to fluoride abstraction reactions in methanol, leading to dinuclear fluoride-bridged uranyl(VI) complexes. Fluoride abstraction of 2b in methanol generates two structural isomers of the fluoride-bridged uranyl(VI) dimer [(UO2(dppmo)2)2(mu-F)][BF4]3 (4), both of which have been structurally characterized. In the major isomer 4C, the four dppmo ligands are all chelating, while in the minor isomer 4B, two of the dppmo ligands bridge adjacent uranyl(VI) centers. Photolysis of 2b in methanol proceeds through 4 to form the uranium(IV) fluoride complex [UO2F2(dppmo)3][BF4]2 (5), involving another fluoride abstraction step. X-ray crystallography shows 5 to be a rare example of a structurally characterized uranium(IV) complex possessing terminal U-F bonds. Complex 5 reverts to 4 in solution upon exposure to air. PMID- 17083219 TI - Mononuclear Schiff base boron halides: synthesis, characterization, and dealkylation of trimethyl phosphate. AB - A series of mononuclear boron halides of the type LBX(2) [LH = N-phenyl-3,5-di tert-butylsalicylaldimine, X = Cl (2), Br (3)] and LBX [LH2 = N-(2-hydroxyphenyl) 3,5-di-tert-butylsalicylaldimine, X = Cl (7), Br (8); LH2 = N-(2-hydroxyethyl) 3,5-di-tert-butylsalicylaldimine, X = Cl (9), Br (10); and LH2 = N-(3 hydroxypropyl)-3,5-di-tert-butylsalicylaldimine, X = Cl (11), Br (12)] were synthesized from their borate precursors LB(OMe)2 (1) (LH = N-phenyl-3,5-di-tert butylsalicylaldimine) and LB(OMe) [LH2 = N-(2-hydroxyphenyl)-3,5-di-tert butylsalicylaldimine (4), N-(2-hydroxyethyl)-3,5-di-tert-butylsalicylaldimine (5), N-(3-hydroxypropyl)-3,5-di-tert-butylsalicylaldimine (6)]. The boron halide compounds were air and moisture sensitive, and upon hydrolysis, compound 7 resulted in the oxo-bridged compound 13 that contained two seven-membered boron heterocycles. The boron halide compounds dealkylated trimethyl phosphate in stoichiometric reactions to produce methyl halide and unidentified phosphate materials. Compounds 8 and 12 were found to be the most effective dealkylating agents. On reaction with tert-butyl diphenyl phosphinate, compound 8 produced a unique boron phosphinate compound LB(O)OPPh2 (14) containing a terminal phosphinate group. Compounds 1-14 were characterized by 1H, 13C, 11B, 31P NMR, IR, MS, EA, and MP. Compounds 5, 6, and 11-14 also were characterized by single crystal X-ray diffraction. PMID- 17083221 TI - Coordination framework hosts consisting of 4-pyridyl-substituted carboxylic acid (PCA) dimers and 1D chains of Ni2+ and SCN-: a rational structural extension toward coordination framework hosts with large rectangular cavities. AB - For the expansion of a rectangular cavity (RC) defined by two isonicotinic acid (isoH) dimers as bridging ligands and two SCN bridges, we conducted a structural extension based on the elongation of the bridging ligands by the replacement of isoH with longer 4-pyridyl-substituted carboxylic acid (PCA). For this purpose, the following three PCAs have been employed: trans-3-(4-pyridyl)propenoic acid (acrylH), 4-(4-pyridyl)benzoic acid (pybenH), and trans-3-(4-(4 pyridyl)phenyl)propenoic acid (pppeH). Self-assembly of Ni2+, SCN-, and each of four PCAs involving isoH, acrylH, pybenH, and pppeH in the presence of an aromatic guest gave four inclusion compounds formulated as [Ni(SCN)2(isoH)2].1/2(benz[a]anthracene) (1), [Ni(SCN)2(acrylH)2].1/2(benz[a]anthracene) (2), [Ni(SCN)2(pybenH)2].(pyrene) (3), and [Ni(SCN)2(pppeH)2](3/)(2).(benz[a]anthracene) (4). X-ray crystal structural determination of 1-4 revealed that the proposed structural extension was successful. Their crystal structures are layered structures of two-dimensional (2D) grid-type coordination frameworks (2D host layers) framed with bridging ligands of the corresponding PCA dimers and 1D chains consisting of Ni2+ ions and mu(1,3)-SCN- ions. The lengths of the PCA dimers are 12.269(5) A (isoH dimer), 16.890(4) A (acrylH dimer), 20.89(2) A (pybenH dimer), 25.387(3) A (pppeH dimer A), and 25.527(4) A (pppeH dimer B). Each 2D host layer has RCs defined by the two corresponding PCA dimers and the two SCN bridges. The dimensions of RCs are expanded in proportion to the increase in the lengths of the PCA dimers: 29.52 x 5.60-7.20 A2 (4) > 24.95 x 5.46-7.38 A2 (3) > 20.88 x 5.49-7.25 A2 (2) > 16.41 x 5.53-7.43 A2 (1). These expansions reflect the number of aromatic guests that can be included in RCs. RC of 1 include only one molecule of benz[a]anthracene, whereas RCs of 3 or 4 includes two molecules of pyrene or benz[a]anthracene, respectively. Comparison of the lengths between the PCA dimers and 4,4' bipyridine-type ligands demonstrated that a design strategy-the preparation of a bridging ligand through self-assembly of two PCAs-is both efficient and particularly suitable for the preparation of very long bridging ligands. PMID- 17083220 TI - Environmental effects on the structure of metal ion-DOTA complexes: an ab initio study of radiopharmaceutical metals. AB - Quantum chemical calculations were performed to study the differences between the important radiopharmaceutical metals yttrium (Y) and indium (In) bound by DOTA and modified DOTA molecules. Energies were calculated at the MP2/6-31+G(d)//HF/6 31G(d) levels, using effective core potentials on the Y and In ions. Although the minimum energy structures obtained are similar for both metal ion-DOTA complexes, changes in coordination and local environment significantly affect the geometries and energies of these complexes. Coordination by a single water molecule causes a change in the coordination number and a change in the position of the metal ion in In-DOTA, but Y-DOTA is hardly affected by water coordination. When one of the DOTA carboxylates is replaced by an amide, the resulting structures show a large variation between the Y and In ions. A six-residue model of the active site containing metal ion-DOTA showed that the Y-DOTA structure optimized to a structure similar to the crystal structure but that the water molecule in In-DOTA disrupts the salt bridge between Arg98B and a carboxylate side chain of DOTA. These observed differences could in part explain the differential binding constants for Y-DOTA and In-DOTA to the antibody 2D12.5. PMID- 17083222 TI - Efficient green-blue-light-emitting cationic iridium complex for light-emitting electrochemical cells. AB - A highly luminescent novel cationic iridium complex [iridium bis(2 phenylpyridine)(4,4'-(dimethylamino)-2,2'-bipyridine)]PF6 was synthesized and characterized using NMR, UV-visible absorption, and emission spectroscopy and electrochemical methods. This complex displays intense photoluminescence maxima in the green-blue region of the visible spectrum and exhibits unprecedented phosphorescence quantum yields, 80 +/- 10% with an excited-state lifetime of 2.2 mus in a dichloromethane solution at 298 K. Single-layer light-emitting electrochemical cells with the charged complex as conducting and electroluminescent material sandwiched between indium-tin oxide and Ag electrodes were fabricated, which emit green-blue light with an onset voltage as low as 2.5 V. Density functional theory calculations were performed to provide insight into the electronic structure of the [iridium bis(2-phenylpyridine)(4,4' (dimethylamino)-2,2'-bipyridine)]PF6 complex, comparing these results with those obtained for [iridium bis(2-phenylpyridine)(4,4'-tert-butyl-2,2'-bipyridine)]PF6. PMID- 17083223 TI - Synthesis and structure of new water-soluble and stable tantalum compound: ammonium tetralactatodiperoxo-mu-oxo-ditantalate(V). AB - The stable water-soluble tantalum complex with lactic acid (ammonium tetralactatodiperoxo-mu-oxo-ditantalate(V)), (NH4)4[Ta2(C3H4O3)4(O2)2O].3H2O, was prepared in the crystalline form. According to the single-crystal X-ray diffraction data, this compound forms a monoclinic cell with a = 13.85(2) A, b = 9.06(1) A, c = 12.32(2) A, and beta = 116.30 degrees , space group C2 (No. 2), and has Z = 2 molecules per unit cell. The solid-state 13C NMR data and low flack parameter are consistent with the determined structure. Appearance of the same vibration modes in Raman and IR spectra supports the choice of the space group without inversion symmetry. The solution of the tantalum complex was successfully applied for the synthesis of two photocatalytic materials, NaTaO3 and Sr2Ta2O7. PMID- 17083224 TI - Spontaneously resolved homochiral 3D lanthanide-silver heterometallic coordination framework with extended helical Ln-O-Ag subunits. AB - Two novel homochiral lanthanide-silver heterometallic coordination polymers LnAg(OAc)(IN)3 [Ln = Nd (1), Eu (2), HIN = isonicotinic acid, HOAc = acetic acid] have been prepared under hydrothermal conditions, which were characterized by elemental analysis, infrared, thermogravimetric analysis, and single-crystal X ray diffraction. Both complexes are isostructural and crystallize in a hexagonal system, chiral space group P6(1)22. Both polymers are constructed from infinite right-handed homochiral helical chains with Ln-O-Ag connectivity, representing the first examples of homochiral lanthanide-transition metal heterometallic coordination polymers with a 3D coordination framework based on spontaneous resolution. Furthermore, the luminescent properties of 2 were studied. PMID- 17083225 TI - Methoxy-substituted TQEN family of fluorescent zinc sensors. AB - Two methoxy-substituted TQEN (N,N,N',N'-tetrakis(2 quinolylmethyl)ethylenediamine) derivatives, T(MQ)EN (N,N,N',N'-tetrakis(6 methoxy-2-quinolylmethyl)ethylenediamine) and T(TMQ)EN (N,N,N',N'-tetrakis(5,6,7 trimethoxy-2-quinolylmethyl)ethylenediamine), have been prepared, and their fluorescence properties with respect to Zn2+ coordination were investigated. Introduction of a methoxy substituent at 6-position of the quinoline ring enhances the fluorescence intensity by 10-fold, and the three methoxy substituents in the 5,6,7-positions afford significant enhancement of the long wavelength component of the fluorescence of zinc complex. The substituents did not alter the binding affinity of these compounds toward zinc ion significantly. T(MQ)EN was proved to be effective in detection of zinc ion in cells by fluorescent microscopy. PMID- 17083226 TI - Equilibrium and formation/dissociation kinetics of some Ln(III)PCTA complexes. AB - The protonation constants () of 3,6,9,15-tetraazabicyclo[9.3.1]pentadeca 1(15),11,13-triene-3,6,9-triacetic acid (PCTA) and stability constants of complexes formed between this pyridine-containing macrocycle and several different metal ions have been determined in 1.0 M KCl at 25 degrees C and compared to previous literature values. The first protonation constant was found to be 0.5-0.6 log units higher than the value reported previously, and a total of five protonation steps were detected (log = 11.36, 7.35, 3.83, 2.12, and 1.29). The stability constants of complexes formed between PCTA and Mg2+, Ca2+, Cu2+, and Zn2+ were also somewhat higher than those previously reported, but this difference could be largely attributed to the higher first protonation constant of the ligand. Stability constants of complexes formed between PCTA and the Ln3+ series of ions and Y3+ were determined by using an "out-of-cell" potentiometric method. These values ranged from log K = 18.15 for Ce(PCTA) to log K = 20.63 for Yb(PCTA), increasing along the Ln series in proportion to decreasing Ln3+ cation size. The rates of complex formation for Ce(PCTA), Eu(PCTA), Y(PCTA), and Yb(PCTA) were followed by conventional UV-vis spectroscopy in the pH range 3.5 4.4. First-order rate constants (saturation kinetics) obtained for different ligand-to-metal ion ratios were consistent with the rapid formation of a diprotonated intermediate, Ln(H(2)PCTA)(2+). The stabilities of the intermediates as determined from the kinetic data were 2.81, 3.12, 2.97, and 2.69 log K units for Ce(H(2)PCTA), Eu(H(2)PCTA), Y(H(2)PCTA), and Yb(H(2)PCTA), respectively. Rearrangement of these intermediates to the fully chelated complexes was the rate determining step, and the rate constant (k(r)) for this process was found to be inversely proportional to the proton concentration. The formation rates (k(OH)) increased with a decrease in the lanthanide ion size [9.68 x 10(7), 1.74 x 10(8), 1.13 x 10(8), and 1.11 x 10(9) M(-1) s(-1) for Ce(PCTA), Eu(PCTA), Y(PCTA), and Yb(PCTA), respectively]. These data indicate that the Ln(PCTA) complexes exhibit the fastest formation rates among all lanthanide macrocyclic ligand complexes studied to date. The acid-catalyzed dissociation rates (k1) varied with the cation from 9.61 x 10(-4), 5.08 x 10(-4), 1.07 x 10(-3), and 2.80 x 10(-4) M(-1) s(-1) for Ce(PCTA), Eu(PCTA), Y(PCTA), and Yb(PCTA), respectively. PMID- 17083227 TI - Effect of the environment on molecular properties: synthesis, structure, and photoluminescence of Cu(I) bis(2,9-dimethyl-1,10-phenanthroline) nanoclusters in eight different supramolecular frameworks. AB - By selection of different charge-balancing anionic frameworks and different host to-guest ratios, the photosensitizer-dye cation [Cu(dmp)2]+ (dmp = 2,9-dimethyl 1,10-phenanthroline) has been embedded in a series of three-dimensional host structures. It occurs with variable geometry in different states of aggregation, including weakly interacting monomers, isolated dimers, columns, and layers. A large variation in its emission lifetime is correlated with the relative energy level spacings of the guest- and host-framework components. In a fully saturated host framework, the lifetime exceeds values reported for a series of conventional Cu(dmp)2 salts. PMID- 17083228 TI - Flexible and shape-selective guest binding at Cu(II) axial sites in 1-dimensional Cu(II)-1,2-bis(4-pyridyl)ethane coordination polymers. AB - A series of guest-binding Cu(II) coordination polymers, {[Cu(bpetha)2(acetone)2].2PF6}n (bpetha = 1,2-bis(4-pyridyl)ethane) (1), {[Cu(bpetha)2(DMF)2].2PF6}n (2), {[Cu(bpetha)(2)(MeCN)(2)].2PF6.2MeCN}n (3), {[Cu(bpetha)2(H2O)2].2PF6.3THF.2H2O}n (4), {[Cu(bpetha)2(H2O)2].2PF6.3dioxane}n (5), and {[Cu(bpetha)2(H2O)2].2PF6.2-PrOH.2H2O}n (6), have been synthesized and crystallographically characterized. Their framework stabilities and guest exchange properties have also been investigated. All compounds form a similar framework motif, a "double chain", in which the bpetha ligands bridge Cu(II) centers to form 1-D [Cu(bpetha)2]n double chains. A variety of Lewis base guest molecules, such as H2O, acetone, DMF, MeCN, THF, dioxane, and 2-PrOH, are incorporated into the assembly of the 1-D double chains. These chains flexibly change their forms of assembly in a guest-dependent manner. Interestingly, acetone, DMF, and MeCN guests with a carbonyl or cyanide group coordinate directly to the axial sites of the Cu(II) centers; in contrast, THF, dioxane, and 2-PrOH guests with an ether or alcohol group are incorporated into the frameworks not via coordination bonds but via weak interactions (hydrogen bonds and van der Waals forces). This selectivity is probably due to steric effects at coordinated oxygen or nitrogen atoms of the guests. Crystal-to-crystal transformations triggered by guests are observed, during which guests coordinated to the Cu(II) axial sites are readily removed and replaced by other guests. PMID- 17083229 TI - NH4[BGe3O8]: a new borogermanate framework made of infinite-chain building blocks. AB - A new microporous borogermanate NH4[BGe3O8] has been synthesized by a molten boric acid flux method with "reagent" quantities of water in which GeO2, ethylenediamine, H2O, and H3BO3 (5:8:14:25) were heated together at 513 K for 4 days. The structure consists of {Ge6O18}n chains, further linked together via BO4 tetrahedra, forming a three-dimensional open framework with intersecting channel systems including one-dimensional 10-membered-ring (MR) channels. Interestingly, the infinite chains {Ge6O18}n as building blocks, built of alternating 4- and 6 MRs made of vertex-sharing GeO4 tetrahedra, construct the borogermanate framework. It is noteworthy that the high viscosity of the reactive medium and the quantity of water play important roles in the formation of the compound. PMID- 17083230 TI - Complexes of greatly enhanced thermodynamic stability and metal ion size-based selectivity, formed by the highly preorganized non-macrocyclic ligand 1,10 phenanthroline-2,9-dicarboxylic acid. A thermodynamic and crystallographic study. AB - The metal ion-complexing properties of 1,10-phenanthroline-2,9-dicarboxylic acid (PDA) are reported. The protonation constants (pK1 = 4.75, pK2 = 2.53) and formation constants (log K(1)) for PDA with Mg(II) (3.53), Ca(II) (7.3), Sr(II) (5.61), Ba(II) (5.43), La(III) (13.5), Gd(III) (16.1), Zn(II) (11.0), Cd(II) (12.8), Pb(II) (11.4), and Cu(II) (12.8) were determined by UV-vis spectroscopy in 0.1 M NaClO4 at 25 degrees C. The log K(1) values for most of these metal ions were high enough that they were not displaced from their PDA complexes even at pH 2. The log K(1) values were determined using the UV spectra to monitor the competition with EDTA (or DTPA; EDTA = ethylendiamine tetraacetic acid, DTPA = diethylenetriamine pentaacetic acid) as a function of pH according to the equilibrium: M(EDTA) + PDA + nH+ = M(PDA) + EDTAHn. The log K1 values indicate that the rigid extended aromatic backbone of PDA leads to high levels of ligand preorganization and selectivity toward large metal ions (e.g., Ca(II), Cd(II), Gd(III)) with an ionic radius of about 1.0 A and greatly enhanced thermodynamic stability as compared to similar ligands without the reinforcing aromatic backbone. The structure of [Ca(PDA)(H2O)2].2H2O (1) is reported: orthorhombic, Fdd2, a = 44.007(9) A, b = 18.945(4) A, c = 7.2446(14) A, V = 6040(2) A(3), Z = 16, R = 0.0882. The Ca(II) ion has a coordination number of eight, lying in the plane of the tetradentate PDA, with Ca-N bonds averaging 2.55 A and Ca-O bonds to the two acetate groups of PDA averaging 2.45 A. These are very close to the normal Ca-L bonds of this type, supporting the idea that a metal ion the size of Ca(II) (ionic radius approximately 1.0 A) will fit into PDA in a low-strain manner. The remaining four coordination sites on Ca(II) in 1 come from two coordinated water molecules and a chelating carboxylate bridging from an adjacent [Ca(PDA)(H2O)2].2H2O complex. Potential applications of PDA as a ligand in biomedical applications such as Gd(III) contrast agents in MRI are discussed. PMID- 17083231 TI - Heterobimetallic Zn(II)-Ln(III) phenylene-bridged schiff base complexes, computational studies, and evidence for singlet energy transfer as the main pathway in the sensitization of near-infrared Nd3+ luminescence. AB - A series of 3d-4f heterobimetallic phenylene-bridged Schiff base complexes of the general formula [Zn(mu-L1)Ln(NO3)3(S)n] [Ln = La (1), Nd (2), Gd (3), Er (4), Yb (5); S = H(2)O, EtOH; n = 1, 2; H2L1 = N,N'-bis(3-methoxysalicylidene)phenylene 1,2-diamine] and [Zn(mu-L2)Ln(NO3)3(H2O)n] [Ln = La (6), Nd (7), Gd (8), Er (9), Yb (10); n = 1, 2; H(2)L(2) = N,N'-bis(3-methoxy-5-p-tolylsalicylidene)phenylene 1,2-diamine] were synthesized and characterized. Complexes 1, 2, 4, and 7 were structurally characterized by X-ray crystallography. At room temperature in CH(3)CN, both neodymium(III) (2 and 7) and ytterbium(III) (5 and 10) complexes also exhibited, in addition to the ligand-centered emission in the UV-vis region, their lanthanide(III) ion emission in the near-infrared (NIR) region. The photophysical properties of the zinc(II) phenylene-bridged complexes (ZnL1 and ZnL2) were measured and compared with those of the corresponding zinc(II) ethylene-bridged complexes (ZnL3 and ZnL4). Our results revealed that, at 77 K, both ligand-centered triplet (3LC) and singlet (1LC) states existed for the ethylene-bridged complexes (ZnL3 and ZnL4), whereas only the (1)LC state was detected for the phenylene-bridged complexes (ZnL1 and ZnL2). NIR sensitization studies of [Zn(mu-L')Nd(NO3)3(H2O)n] (L' = L1-L4) complexes further showed that Nd3+ sensitization took place via the 3LC and 1LC states when the spacer between the imine groups of the Schiff base ligand was an ethylene and a phenylene unit, respectively. Ab initio calculations show that the observed differences can be attributed to the difference in the molecular vibrational properties and electron densities of the electronic states between the ethylene- and phenylene-bridged complexes. PMID- 17083232 TI - Crystal structure, Judd-Ofelt analysis, and spectroscopic assessment of a TmAl3(BO3)4 crystal as a new potential diode-pumped laser near 1.9 micron. AB - TmAl3(BO3)4 crystallizes in the trigonal system R32 (No. 155) with a = b = 9.2741(13) A, c = 7.218(3) A, alpha = beta = 90 degrees , gamma = 120 degrees , V = 537.7(2) A(3), D(c) = 4.494 g cm(-3), and Z = 3. The absorption spectrum of this crystal was recorded at room temperature. The Judd-Ofelt (J-O) theory was applied to the absorption intensities of TmAl(3)(BO3)4 to obtain the three J-O parameters: Omega(2) = 2.40 x 10(-20) cm(2), Omega(4) = 0.48 x 10(-20) cm(2), and Omega(6) = 1.09 x 10(-20) cm(2). The radiative probabilities, radiative lifetimes, and branching ratios of TmAl3(BO3)4 were calculated. The absorption and emission cross sections, together with the potential laser gain near 1.9 microm, were investigated. The potential laser gain curves indicate that the tunability range is about 200 nm. PMID- 17083233 TI - On the role of the bridging dicyanamidobenzene ligand in a new binuclear ruthenium complex: [{Ru(tpy)(thd)}2(mu-dicyd)][PF6] with tpy = 2,2':6',2' ' terpyridine and thd = 2,2,6,6-tetramethyl-3,5-heptanedione. AB - The dicyanamidobenzene-bridge diruthenium complex [{Ru(tpy)(thd)}(2)(mu dicyd)][PF(6)] ([3][PF(6)]) (dicyd = 1,4-dicyanamidobenzene, tpy = 2,2':6',2' ' terpyridine, thd = 2,2,6,6-tetramethyl-3,5-heptanedione) and its mononuclear counterpart [Ru(tpy)(thd)(Ipcyd)] (2) [Ipcyd = 4-iodophenylcyanamide anion (Ipcyd(-))] were synthesized and fully characterized. Cyclic voltammetry of 3 showed the presence of four reversible one-electron redox couples. UV-vis-NIR spectroelectrochemistry and EPR spectroscopy of the electrogenerated paramagnetic intermediates were used to ascertain the oxidation-state distribution. The stable starting dinuclear complex 3(+) is found to be a ligand-centered anion radical as shown by EPR spectroscopy, magnetic susceptibility measurements, and DFT calculations. Oxidation of 3(+) to 3(2+) led to an EPR silent system due to substantial intramolecular antiferromagnetic interaction of the electron spins carried by the low spin ruthenium(III) atom and the bridging anion radical dicyanamido (dicyd(*)(-)), an observation which was supported by UV-vis-NIR, X ray structure, and DFT calculations. Complex 3(3+) presented an EPR spectra consistent with a total effective spin S = (1)/(2) issued from an antiferromagnetic interaction of electron spins carried by two low spin ruthenium(III) atoms and the bridging anion radical dicyd(*)(-) in accordance with UV-vis-NIR. This study shows that the dicyanamidobenzene bridging ligand has indubitably a noninnocent behavior. PMID- 17083234 TI - Solid-state coordination chemistry of the Cu/triazolate/X system (X = F-, Cl-, Br , I-, OH-, and SO4(2-)). AB - Hydrothermal reactions of 1,2,4-triazole with the appropriate copper salt have provided eight structurally unique members of the Cu/triazolate/X system, with X = F-, Cl-, Br-, I-, OH-, and SO4(2-). The anionic components X of [Cu3(trz)4(H2O)3]F2 (1) and [Cu6(trz)4Br]Cu4Br4(OH) (4) do not participate in the framework connectivity, acting as isolated charge-compensating counterions. In contrast, the anionic subunits X of [Cu(II)Cu(I)(trz)Cl2] (2), [Cu6(trz)4Br2] (3), [Cu(II)Cu(I)(trz)Br2] (5), [Cu3(trz)I2] (6), [Cu6(II)Cu2(I)(trz)6(SO4)3(OH)2(H2O)] (8), and [Cu4(trz)3]OH.7.5H2O (9.7.5H2O) are intimately involved in the three-dimensional connectivities. The structure of [Cu(II)Cu(I)(trz)2][Cu3(I)I4] (7) is constructed from two independent substructures: a three-dimensional cationic {Cu2(trz)2}n(n+) component and {Cu3I4}n(n-) chains. Curiously, four of the structures are mixed-valence Cu(I)/Cu(II) materials: 2, 5, 7, and 8. The only Cu(II) species is 1, while 3, 4, 6, and 9.7.5H2O exhibit exclusively Cu(I) sites. The magnetic properties of the Cu(II) species 1 and of the mixed-valence materials 5, 7, 8, and the previously reported [Cu3(trz)3OH][Cu2Br4] have been studied. The temperature-dependent magnetic susceptibility of 1 conforms to a simple isotropic model above 13 K, while below this temperature, there is weak ferromagnetic ordering due to spin canting of the antiferromagnetically coupled trimer units. Compounds 5 and 7 exhibit magnetic properties consistent with a one-dimensional chain model. The magnetic data for 8 were fit over the temperature range 2-300 K using the molecular field approximation with J = 204 cm(-1), g = 2.25, and zJ' = -38 cm( 1). The magnetic properties of [Cu3(trz)3OH][Cu2Br4] are similar to those of 8, as anticipated from the presence of similar triangular {Cu3(trz)3(mu3-OH)}(2+) building blocks. The Cu(I) species 3, 4, 6, and 9 as well as the previously reported [Cu(5)(trz)3Cl2] exhibit luminescence thermochromism. The spectra are characterized by broad emissions, long lifetimes, and significant Stokes' shifts, characteristic of phosphorescence. PMID- 17083235 TI - Conjugated metallopolymers for fluorescent turn-on detection of nitric oxide. AB - A series of eight pi-conjugated polymers (CPs) composed of phenylenevinylene, phenyleneethynylene, fluorene, and thiophene derivatives have been prepared with bipyridyl or terpyridyl substituents within the pi-conjugated backbone or at side chain positions. These ligand-modified CPs serve as macromolecular scaffolds for conducting metallopolymers. The optical and photoluminescent properties of the polymers and corresponding copper(II) metallopolymers were investigated. Copper(II) is a highly efficient quencher of CP emission (75-100% quenching). CPs featuring bipyridyl units within the CP backbone are quenched more efficiently than those with terpyridyl units. The copper(II) metallopolymer undergoes reduction to the corresponding copper(I) species upon reaction with nitric oxide, with concomitant changes in integrated emission ranging from a 50% decrease to a 320% increase. The positive emission response is largest when Cu(II) was bound to the CP through bipyridyl units within the backbone, making these materials the best candidates for NO sensing by a turn-on emission mechanism. PMID- 17083236 TI - Crystal structures and 77Se NMR spectra of molybdenum(IV) areneselenolates having intramolecular NH...Se hydrogen bonds. AB - Salts of the monooxomolybdenum(IV,V) areneselenolates having intramolecular NH...Se hydrogen bonds, [Mo(IV)O(Se-2-RCONHC6H4)4]2- (R = t-Bu, CH3, CF3) and [Mo(V)O(Se-2-t-BuCONHC6H4)4]-, were synthesized and characterized by 1H nuclear magnetic resonance (NMR), 77Se NMR, electron spin resonance (ESR), UV-visible spectra, X-ray analysis, and electrochemical measurements. 77Se-1H correlated spectroscopy (COSY) indicated a significant correlation between amide 1H and selenolate 77Se atoms through an NH...Se hydrogen bond with 1J(77Se-1H) = 5.4 Hz coupling. The hydrogen bonds contribute to the positive shift in the Mo(V)/Mo(IV) redox potential. In the crystal structure of (PPh4)2[Mo(IV)O(Se-2-CH3CONHC6H4)4], an NH...O=Mo hydrogen bond was found. Ab inito calculations support the presence of intramolecular NH...O=Mo and NH...Se hydrogen bonds. PMID- 17083237 TI - Structure and thermoelectric characterization of Ba8Al14Si31. AB - A molten Al flux method was used to grow single crystals of the type I clathrate compound Ba8Al14Si31. Single-crystal neutron diffraction data for Ba8Al14Si31 were collected at room temperature using the SCD instrument at the Intense Pulsed Neutron Source, Argonne National Laboratory. Single-crystal neutron diffraction of Ba8Al14Si31 confirms that the Al partially occupies all of the framework sites (R1 = 0.0435, wR2 = 0.0687). Stoichiometry was determined by electron microprobe analysis, density measurements, and neutron diffraction analysis. Solid-state (27)Al NMR provides additional evidence for site preferences within the framework. This phase is best described as a framework-deficient solid solution Ba8Al14Si31, with the general formula, Ba(8)Al(x)Si(42-3/4x)[](4-1/4x) ([] indicates lattice defects). DSC measurements and powder X-ray diffraction data indicate that this is a congruently melting phase at 1416 K. Temperature dependent resistivity reveals metallic behavior. The negative Seebeck coefficient indicates transport processes dominated by electrons as carriers. PMID- 17083238 TI - Electronic structure and charge transfer in the ternary intercalated graphite beta-KS0.25C3. AB - The electronic structure of the ternary intercalated graphite beta-KS(0.25)C3 is studied by means of a first-principles density functional theory approach. The nature of the partially filled bands is analyzed, and the K sublayers of the intercalate are shown to have an important contribution to the Fermi surface. This K-based contribution confers a sizable three-dimensional character to the conductivity even if considerably less than that for the related binary KC8. The electronic structure of beta-KS(0.25)C3 differs noticeably from that of the related ternary compound, KH(x)C4. The charge transfer is analyzed, and a way to evaluate it, which can be used in general for intercalated graphites, is proposed. The charge transfer per C atom in this ternary material is shown to be smaller than that in the KC8 binary compound despite a more favorable stoichiometry ratio between K and C. PMID- 17083239 TI - Ferrocenyldiselenolate-stabilized copper-selenium clusters. AB - The silylated ferrocenyl selenium reagent 1,1'-Fe(eta5-C5H4SeSiMe3)2 has been used for the high yield formation of the phosphine-ligated copper complexes Cu2(fcSe2)(PiPr3)2 (1) and Cu4(fcSe2)2(PnPr3)4 (2) from solublilized CuOAc, as determined by single-crystal X-ray diffraction. The incorporation of a source of Se2- into the reaction scheme with the reagent Se(SiMe3)2 yields the mixed selenide/ferrocenyldiselenolate cluster [Cu20Se6(Se2fc)4(PnPr3)10] (3). Partial substitution of the PnPr3 ligand shell in 3 with the phosphinothiol Ph2P(CH2)3SH leads to an expansion of the framework and the high yield formation of the crystallographically characterized cluster Cu36(fcSe2)6Se12(PnPr3)10(Ph2P(CH2)3SH)2 (5), which contains surface alkylthiol groups on a copper-selenium core. PMID- 17083240 TI - Synthesis of 1D {Cu6(mu3-SC3H6N2)4(mu-SC3H6N2)2(mu-I)2I4}n and 3D {Cu2(mu SC3H6N2)2(mu-SCN)2}n polymers with 1,3-imidazolidine-2-thione: bond isomerism in polymers. AB - The reaction of copper(I) iodide with 1, 3-imidazolidine-2-thione (SC3H6N2) in a 1:2 molar ratio (M/L) has formed unusual 1D polymers, {Cu6(mu3-SC3H6N2)4(mu SC3H6N2)2(mu-I)2I4}n (1) and {Cu6(mu3-SC3H6N2)2(mu-SC3H6N2)4(mu-I)4I2}n (1a). A similar reaction with copper(I) bromide has formed a polymer {Cu6(mu3 SC3H6N2)2(mu-SC3H6N2)4(mu-Br)4Br2}n (3a), similar to 1a, along with a dimer, {Cu2(mu-SC3H6N2)2(eta1-SC3H6N2)2Br2} (3). Copper(I) chloride behaved differently, and only an unsymmetrical dimer, {Cu2(mu-SC3H6N2)(eta1-SC3H6N2)3Cl2} (4), was formed. Finally, reactions of copper(I) thiocyanate in 1:1 or 1:2 molar ratios yielded a 3D polymer, {Cu2(mu-SC3H6N2)2(mu-SCN)2}n (2). Crystal data: 1, C9H18Cu3I3N6S3, triclinic, P, a = 9.6646(11) A, b = 10.5520(13) A, c = 12.6177(15) A, alpha = 107.239(2) degrees , beta = 99.844(2) degrees , gamma = 113.682(2) degrees , V = 1061.8(2) A(3), Z = 2, R = 0.0333; 2, C(4)H(6)CuN(3)S(2), monoclinic, P2(1)/c, a = 7.864(3) A, b = 14.328(6) A, c = 6.737(2) A, beta = 100.07(3) degrees , V = 747.4(5), Z = 4, R = 0.0363; 3, C12H24Br2Cu2N8S4, monoclinic, C2/c, a = 19.420(7) A, b = 7.686(3) A, c = 16.706(6) A, beta = 115.844(6) degrees , V = 2244.1(14) A(3), Z = 4, R = 0.0228; 4, C12H24Cl2Cu2N8S4, monoclinic, P2(1)/c, a = 7.4500(6) A, b = 18.4965(15) A, c = 16.2131(14) A, beta = 95.036(2) degrees , V = 2225.5(3) A(3), Z = 4, R = 0.0392. The 3D polymer 2 exhibits 20-membered metallacyclic rings in its structure, while synthesis of linear polymers, 1 and 1a, represents an unusual example of I (1a)-S (1) bond isomerism. PMID- 17083241 TI - Deep-red luminescence and efficient singlet oxygen generation by cyclometalated platinum(II) complexes with 8-hydroxyquinolines and quinoline-8-thiol. AB - The synthesis and photophysical study of (C/?N)Pt(II)Q complexes, where C/?N is a bidentate cyclometalating ligand and Q is 8-hydroxyquinoline or quinoline-8 thiol, are presented. The compounds were obtained as a single isomer with N atoms of the C/?N and Q ligands trans-coordinated to the Pt(II) center as shown by X ray crystallography. These chromophores absorb intensely in the visible region and emit in the deep-red spectral region from a quinolate-centered triplet intraligand charge-transfer excited state. The emission maxima are in the range 675-740 nm, with the quantum yields and lifetimes of up to 0.82% and 5.3 mus, respectively, in deoxygenated organic solvents at room temperature. These complexes are efficient photosensitizers of singlet oxygen in air-saturated solutions, with yields up to 90%. PMID- 17083242 TI - Copper(I) coordination chemistry of (pyridylmethyl)amide ligands. AB - Copper(I) chloro complexes were synthesized with a family of ligands, HL(R) [HL(R) = N-(2-pyridylmethyl)acetamide, R = null; 2-phenyl-N-(2 pyridylmethyl)acetamide, R = Ph; 2,2-dimethyl-N-(2-pyridylmethyl)propionamide, R = Me3; 2,2,2-triphenyl-N-(2-pyridylmethyl)acetamide, R = Ph3)]. Five complexes were synthesized from the respective ligand and cuprous chloride: [Cu(HL)Cl]n (1), [Cu2(HL)4Cl2] (2), [Cu2(HL(Ph))2(CH3CN)2Cl2] (3), [Cu2(HL(Ph)3)2Cl2] (4), and [Cu(HL(Me)3)2Cl] (5). X-ray crystal structures reveal that for all complexes the ligands coordinate to the Cu in a monodentate fashion, and inter- or intramolecular hydrogen-bonding interactions formed between the amide NH group and either amide C=O or chloro groups stabilize these complexes in the solid state and strongly influence the structures formed. Complexes 1-5 display a range of structural motifs, depending on the size of the ligand substituent groups, hydrogen bonding, and the stoichiometry of the starting materials, including a one-dimensional coordination polymer chain (1) and binuclear (2-4) or mononuclear (5) structures. PMID- 17083243 TI - Mechanism of the reactions of synthetic Fe-S-based clusters with PhCOCl: parallel pathways involving free and coordinated thiolate as nucleophiles. AB - Terminal thiolate ligands on the synthetic Fe-S-based clusters [Fe4S4(SR)4]2- (R = Et or SPh) or [{MoFe3S4(SPh)3}2(mu-SPh)3]3- are replaced by chloride in a reaction with PhCOCl to produce [Fe4S4Cl4]2- and [{MoFe3S4Cl3}2(mu-SPh)3]3-, respectively. Kinetic studies using stopped-flow spectrophotometry show that, in general, the mechanisms of these reactions in MeCN occur by two pathways. One pathway is independent of the concentration of PhCOCl and involves rate-limiting dissociation of the thiolate ligand. The free thiolate subsequently reacts with PhCOCl to produce PhCOSR and the Cl- which binds to the vacant site on the cluster. The second pathway exhibits a nonlinear dependence on the concentration of PhCOCl and involves initial, rapid binding of PhCOCl to the cluster followed by intramolecular thiolate ligand attack on the coordinated acid chloride. The intermediate in which PhCOCl is bound to the cluster has been detected spectrophotometrically. The ways in which the rates of the reactions between PhCOCl and Fe-S-based clusters are affected by changes of the terminal thiolate, the metal composition of the cluster core, and the protonation state of the cluster have been investigated and are compared with the effect these same changes have on the rates of nucleophilic substitution. PMID- 17083244 TI - Computational studies on the photophysical properties and NMR fluxionality of the tetranuclear copper(I) complexes [Cu4(mu-dppm)4(mu4-E)]2+ (E = PPh and S). AB - Density functional theory (DFT) calculations at the hybrid Perdew, Burke, and Ernzerhof functional level were performed to study the electronic structures of the ground and excited states of the luminescent tetranuclear copper(I) complexes [Cu4(mu-dppm)4(mu4-E)]2+ [E = PPh (1) and S (2)] by using model complexes [Cu4(mu H2PCH2PH2)4(mu4-E)]2+ [E = PPh (1a) and S (2a)]. The time-dependent DFT method at the same level associated with the conductor-like polarizable continuum model was used to study the nature of the low-energy transitions in their electronic absorption spectra. The results indicate that the lowest energy absorptions of both 1 and 2 are attributed to ligand-to-metal charge-transfer (LMCT) (E --> Cu4) with mixing of metal-cluster-centered (MCC) (3d --> 4s/3d --> 4p) singlet-singlet transitions. The geometry optimizations on the lowest energy triplet state reveal that the emissive states of both complexes involve a considerable structural distortion in which they are derived predominantly from an admixture of 3LMCT (E -> Cu4) and 3MCC (3d --> 4p) origin. In addition to the photophysical properties, the fluxional behavior of 2 observed from the NMR studies but not that of 1 was investigated. It is found that the fluxionality in 2 involves the shuttling of the sulfido ligand through the rectangular Cu4 core. PMID- 17083245 TI - Quantum chemical characterization of the bonding of N-heterocyclic carbenes to Cp2MI compounds [M = Ce(III), U(III)]. AB - The binding of N-heterocyclic carbenes to Ce(III) and U(III) compounds is characterized by quantum chemical methods. Density functional methods are in qualitative agreement with experiment that binding to U(III) is more favorable than to Ce(III); after correcting for basis-set superposition error, quantitative agreement with experiment is achieved with a multireference second-order perturbation theory approach accounting for relativistic effects. The small computed (and observed) preference derives from a combination of several small effects, including differences in electronic binding energies, rovibrational partition functions, and solvation free energies. Prospects for ligand modification to improve the differentiation between lanthanides and actinides are discussed on the basis of computational predictions. PMID- 17083246 TI - 2D-grid layered Pd-based cationic infinite coordination polymer/polyoxometalate crystal with hydrophilic sorption. AB - The inorganic-organic hybrid compound composed of the novel infinite-chain Pd(II) complex and the polyoxometalate ({[(en)Pd(p-bpy)]2[alpha SiW12O40].8DMSO.4DMF}infinity (1a:1 with 8DMSO.4DMF; monoclinic P2(1)/c (No. 14), a = 15.0188(3) A, b = 15.6962(3) A, c = 26.9793(6) A, beta = 106.3580(10) degrees , V = 6102.6(2) A(3)) has been successfully synthesized by the reaction of [(en)Pd(OH2)2]2[alpha-SiW12O40] (2) with 4,4'-bipyridine (p-bpy). The treatment in dry N(2) at 50 degrees C or evacuation at room temperature forms {[(en)Pd(p bpy)](2)[alpha-SiW(12)O(40)].6.0DMSO}(infinity) (1b:1 with 6DMSO) with a drastic reduction of the interlayer spacing, which is characterized by the powder diffraction analysis and the single-crystal analysis of 1c (1 with 4.5DMSO.3.5DMF; monoclinic P2(1)/a (No. 14), a = 14.200(9) A, b = 22.8865(8) A, c = 14.8558(5) A, beta = 114.7990(10) degrees , V = 4383.0(2) A(3)). Compound 1b reversibly sorbs the hydrophilic molecules with the maintenance of the intrinsic structure, which is much different from hydrophobic guest-inclusion properties reported in the other Pd-based supramolecular systems. PMID- 17083247 TI - Synthesis of neutral (Pd(II), Pt(II)), cationic (Pd(II)), and water-induced anionic (Pd(II)) complexes containing new mesocyclic thioether-aminophosphonite ligands and their application in the Suzuki cross-coupling reaction. AB - Mesocyclic thioether-aminophosphonite ligands, {-OC10H6(mu-S)C10H6O-}PNC4H8O (2a, 4-(dinaphtho[2,1-d:1',2'-g][1,3,6,2]dioxathiaphosphocin-4-yl)morpholine) and { OC10H6(mu-S)C10H6O-}PNC4H8NCH3 (2b, 1-(dinaphtho[2,1-d:1',2' g][1,3,6,2]dioxathiaphosphocin-4-yl)-4-methylpiperazine) are obtained by reacting {-OC10H6(mu-S)C10H6O-}PCl (1) with corresponding nucleophiles. The ligands 2a and 2b react with (PhCN)2PdCl2 or M(COD)Cl2 (M = Pd(II) or Pt(II)) to afford P coordinated cis-complexes, [{(-OC10H6(mu-S)C10H6O-)PNC4H8X-kappaP}2MCl2] (3a, M = Pd(II), X = O; 3b, M = Pd(II), X = NMe; 4a, M = Pt(II), X = O; 4b, M = Pt(II), X = NMe). Compounds 2a and 2b, upon treatment with [Pd(eta3-C3H5)Cl]2 in the presence of AgOTf, produce the P,S-chelated cationic complexes, [{(-OC10H6(mu S)C10H6O-)PNC4H8X-kappaP,kappaS}Pd(eta3-C3H5)](CF3SO3) (5a, X = O and 5b, X = NMe). Treatment of 2a and 2b with (PhCN)2PdCl2 in the presence of trace amount of H2O affords P,S-chelated anionic complexes, [{(-OC10H6(mu-S)C10H6O-)P(O) kappaP,kappaS}PdCl2](H2NC4H8X) (6a, X = O and 6b, X = NMe), via P-N bond cleavage. The crystal structures of compounds 1, 2a, 2b, 4a, and 6a are reported. Compound 6a is a rare example of crystallographically characterized anionic transition metal complex containing a thioether-phosphonate ligand. Most of these palladium complexes proved to be very active catalysts for the Suzuki-Miyaura reaction with excellent turnover number ((TON), up to 9.2 x 10(4) using complex 6a as a catalyst). PMID- 17083248 TI - A dinuclear lanthanide complex for the recognition of bis(carboxylates): formation of terbium(III) luminescent self-assembly ternary complexes in aqueous solution. AB - The synthesis and photophysical properties of a coordinatively unsaturated cationic dinuclear terbium complex, 2.Tb(2), that can detect the presence of mono or bis(carboxylates) in buffered aqueous solution at physiological pH is described. Full ligand synthesis and structural characterization of 2.Na(2) are also described. Spectroscopic measurements determined that each Tb(III) metal center has two metal-bound water molecules (q = 2). The recognition or sensing of N,N-dimethylaminocarboxylic acid, 4, and the bis(carboxylate) terephthalic acid, 5, which can also function as sensitizing antennae, was found to occur through the binding of these carboxylates to the metal center via the displacement of the metal bound water molecules. This gave rise to the formation of luminescent ternary complexes in solution in 2:1 or 1:1 (ion:2.Tb(2)) stoichiometry, respectively. Aliphatic bis(carboxylates) also bind to 2.Tb(2) where the selectivity for the ion recognition and stoichiometry was dictated by the structure of the anion, being most selective for pimelic acid, 6. Binding of either l- or d-tartaric acid gave rise to the formation ternary complex formation, with 2:1 stoichiometry, where the ion recognition resulted in quenching of the lanthanide emission. PMID- 17083249 TI - Synthesis and magnetic properties of an amine-templated Fe2+ (s = 2) sulfate with a distorted Kagome structure. AB - An organically templated iron(II) sulfate of the composition [H3N(CH2)2NH2(CH2)2(NH3]4[Fe(II)9F18(SO4)6].9H2O with a distorted Kagome structure has been synthesized under solvothermal conditions in the presence of diethylenetriamine. The distortion of the hexagonal bronze structure comes from the presence of two different types of connectivity between the FeF4O2 octahedra and the sulfate tetrahedra. This compound exhibits magnetic properties different from those of an Fe(II) compound with a perfect Kagome structure and is a canted antiferromagnet at low temperatures. PMID- 17083250 TI - An isomeric pair of fluoride-bridged cyclic dimolybdenum triads. AB - A pair of isomeric cyclic triads containing three quadruply bonded [Mo2] units, [Mo2(cis-DAniF)2]2+ (DAniF = N,N'-di-p-anisylformamidinate), bridged by six fluoride anions, has been synthesized and crystallographically characterized. For the alpha isomer, the three [Mo2] units are oriented in two orthogonal directions. Two of them are structurally equivalent and parallel to each other, but oriented perpendicular to the third one. The beta isomer is a triangle with three geometrically identical [Mo2] units, parallel to each other, as the vertices. Thus, the beta isomer possesses idealized D3h symmetry while the alpha isomer only has C2v symmetry. These two isomers do not interconvert in boiling THF or toluene or under irradiation with ultraviolet light, but oxidation of the alpha isomer first generates an alpha+ species that changes to beta+. The two isomers have very similar electrochemical behavior, both showing three reversible one-electron redox processes for the [Mo2] centers and similar potential separations (DeltaE(1/2)). The first and second redox couples are well separated (ca. 390-410 mV), while the second and third ones are separated by only about 150 mV. PMID- 17083251 TI - Alkali-metalated forms of thiacalix[4]arenes. AB - The alkali metal salts [TCALi4] (1), [TCANa4] (2), and [TCALK4] (3) of fully deprotonated p-tert-butyltetrathiacalix[4]arene (H(4)TCA) are readily available from the reactions of thiacalix[4]arene and n-BuLi, NaH, or KH as deprotonating reagents. Crystals of the sodium salts 2 and the potassium salt 3 suitable for X ray diffraction were obtained in the form of the pyridine solvates [(TCA)2Na8.8py] (2.8py) and [(TCA)2K(8).8py] (3.8py). These molecules are dimers in the solid state but are structurally not related. In addition, the reaction of H(4)TCA and lithium hydroxide afforded the structurally characterized complex [(TCA)Li5(OH).4THF] (4). The molecular structure of 4 as well as the structures of 2.8py and 3.8py reveal a close relationship to the corresponding alkali metal salts of the calix[4]arenes. PMID- 17083252 TI - Strong electronic communication by direct metal-metal interaction in molecules with halide-bridged dimolybdenum pairs. AB - Reactions of [cis-Mo2(DAniF)2(NCCH3)4](BF4)2, DAniF = N,N'-di-p anisylformamidinate, with an excess of anhydrous Bu(n)4NX (X = Cl, Br, I), produced the halide-bridged tetranuclear clusters, [cis-Mo2(DAniF)2]2(mu-X)4, X = Cl (1), Br (2), and I (3). All three compounds show two reversible one-electron oxidation processes with potential separations (DeltaE(1/2)) between the two oxidation processes of 540, 499, and 440 mV, respectively. These DeltaE(1/2) values show that the strength of the electronic coupling between the dimetal units decreases as the Mo2...Mo2 distance increases from 1 to 2, and then to 3. The structures, EPR spectra, and near-IR (NIR) spectra of the corresponding mixed valence species (1-PF6, 2-PF6, and 3-PF6) indicate that the clusters are electronically delocalized. Calculations at the DFT level indicate that the strong electronic communication is principally due to a direct overlap between the delta orbitals from the adjacent dimetal units. PMID- 17083253 TI - Phosphate ester hydrolysis by hydroxo complexes of trivalent lanthanides stabilized by 4-imidazolecarboxylate. AB - The anion of 4-imidazolecarboxylic acid (HL) stabilizes hydroxo complexes of trivalent lanthanides of the type ML(OH)+ (M = La, Pr) and M2L(n)(OH)(6-n) (M = La, n = 2; M = Pr, n = 2, 3; M = Nd, Eu, Dy, n = 1-3). Compositions and stability constants of the complexes have been determined by potentiometric titrations. Spectrophotometric and (1)H NMR titrations with Nd(III) support the reaction model for the formation of hydroxo complexes proposed on the basis of potentiometric results. Kinetics of the hydrolysis of two phosphate diesters, bis(4-nitrophenyl) phosphate (BNPP) and 2-hydroxypropyl 4-nitrophenyl phosphate (HPNPP), and a triester, 4-nitrophenyl diphenyl phosphate (NPDPP), in the presence of hydroxo complexes of five lanthanides were studied as a function of pH and metal and ligand concentrations. With all lanthanides and all substrates, complexes with the smallest n, that is M2L2(OH)4 for La and Pr and M2L(OH)5 for Nd, Eu, and Dy, exhibited the highest catalytic activity. Strong inhibitory effects by simple anions (Cl-, NO3-, (EtO)2PO2-, AcO-) were observed indicating high affinity of neutral hydroxo complexes toward anionic species. The catalytic activity decreased in the order La > Pr > Nd > Eu > Dy for both diester substrates and was practically independent of the nature of cation for a triester substrate. The efficiency of catalysis, expressed as the ratio of the second order rate constant for the ester cleavage by the hydroxo complex to the second order rate constant for the alkaline hydrolysis of the respective substrate, varied from ca. 1 for NPDPP to 10(2) for HPNPP and to 10(5) for BNPP. The proposed mechanism of catalytic hydrolysis involves reversible bridging complexation of a phosphodiester to the binuclear active species followed by attack on the phosphoryl group by bridging hydroxide (BNPP) or by the alkoxide group of the deprotonated substrate (HPNPP). PMID- 17083254 TI - Electrochemistry and homogeneous self-exchange kinetics of the aqueous 12 tungstoaluminate(5-/6-) couple. AB - The effect of alkali metal (M) chloride or triflate supporting electrolytes (0.1 1.0 mol L(-1)) on the midpoint potential E(m) of the aqueous AlW12O40(5-/6-) couple in cyclic voltammetry, after correction (E(corr)) for liquid junction potentials, can be represented in terms of ionic strength according to the extended Debye-Huckel equation. However, unrealistically short AlW12O40(5-/6-) cation closest-approach distances are required to accommodate the specific effects of M+, and the infinite-dilution potential E(corr)(0) values are not quite consistent from one M+ to another. The pressure dependence of Em is qualitatively consistent with expectations based on the Born-Drude-Nernst theory. The strong accelerating effects of supporting electrolytes on the standard electrode reaction rate constant k(el) at pH 3 as measured by alternating current voltammetry (ACV), and on the homogeneous self-exchange rate constant k(ex) at pH 3-7 as measured by 27Al line broadening, depend specifically on the identity and concentration of M+ (Li+ < Na+ < K+ < Rb+) rather than on the ionic strength, whereas the effect of the nature of the supporting anion (Cl- or CF3SO3-) is negligible. Extrapolation of k(el) and k(ex) to zero [M+] indicates that the uncatalyzed electron transfer rate is negligibly small relative to the M+ catalyzed rates. The kinetic effects of M+ show no evidence of the saturation expected had they been due primarily to ion pairing with AlW12O40(5-/6-). The catalytic effect of M+ operates primarily through lowering the enthalpy of activation, which is partially offset by a strongly negative entropy of activation and, for the homogeneous exchange catalyzed by K+ or Rb+, becomes mildly negative; thus, the catalytic effect of M(+) is enthalpy-driven but entropy-limited. For the electrode reaction, the volume of activation averages +4.5 +/- 0.2 cm(3) mol(-1) for all M+ and [M+], in contrast to the negative value predicted theoretically for the uncatalyzed reaction. These results are consistent with a reaction mechanism, previously proposed for other anion-anion electron-transfer reactions, in which anion-anion electron transfer is facilitated by partially dehydrated M+. PMID- 17083255 TI - From monomers to solids: controlled hydrolysis to form novel, heteroleptic hydroxide-containing complexes. AB - Four novel alkaline earth metal tetranuclear aryloxide/pyrazolate hydroxides of the general formula M4(ligand)6(OH)2(donor)n have been prepared by either adding stoichiometric quantities of water to preformed alkaline earth metal aryloxides or in a direct metalation regime combining metal, ligand (alcohol, pyrazole), and donor with stoichiometric amounts of water. The compounds are considered as potential intermediates in the sol-gel process. A nonhydrolyzable magnesium aryloxide has also been obtained using the same reaction scheme. Unexpectedly, a dimeric strontium complex with further association of a second dimer via hydrogen bonding from the solvent of crystallization, ethylenediamine, has been isolated by the treatment of strontium metal with a mixture of alcohol and water. All compounds were characterized using X-ray crystallography, 1H and 13C NMR, and IR spectroscopy. PMID- 17083256 TI - Kinetic effects in heterometallic dinitrogen cleavage. AB - The rhenium(I) dinitrogen complex (PhMe2P)4ClRe(N2) reacts with [Mo2(S2CNEt2)6](OTf)2 (6) to give the N(2)-bridged complex [(PhMe2P)4ClRe(mu N2)Mo(S2CNEt2)3]OTf ([7]OTf). Spectroscopic (nu(NN) = 1818 cm(-1)) and structural data [d(NN) = 1.167(6) A] indicate that the bridging N(2) moiety in 7+ is slightly activated relative to free N2 or to the mononuclear Re complex. However, the complex is stable with respect to N2 cleavage. The putative products of such a cleavage, the known (Et2NCS2)3Mo(N) (5) and the newly prepared [(PhMe2P)(4)ClRe(N)]OTf ([9]OTf), are stable compounds that do not react with each other to give products of nitride coupling. Thus, the failure of 7+ to interconvert with 5 and 9+ is due not to the thermodynamic stability of the NN bond but rather to kinetic factors that disfavor N2 cleavage and nitride coupling. Implications of this result for using polar effects to facilitate N2 cleavage to nitrides as a strategy for nitrogen fixation are discussed. PMID- 17083257 TI - What external perturbations influence the electronic properties of catalase compound I? AB - We have performed density functional theory calculations on an active-site model of catalase compound I and studied the responses of the catalytic center to external perturbations. Thus, in the gas phase, compound I has close-lying doublet and quartet spin states with three unpaired electrons: two residing in pi(FeO) orbitals and the third on the heme. The addition of a dielectric constant to the model changes the doublet-quartet energy ordering but keeps the same electronic configuration. By contrast, the addition of an external electric field along one of the principal axes of the system can change the doublet-quartet energy splitting by as much as 6 kcal mol(-1) in favor of either the quartet or the doublet spin state. This sensitivity is much stronger than the effect obtained for iron heme models with thiolate or imidazole axial ligands. Moreover, an external electric field is able to change the electronic system from a heme based radical [Fe=O(Por*+)OTyr-] to a tyrosinate radical [Fe=O(Por)OTyr*]. This again shows that oxo-iron heme systems are chameleonic species that are influenced by external perturbations and change their character and catalytic properties depending on the local environment. PMID- 17083258 TI - Synthesis, characterization, and reactivity of [Ru(bpy)(CH3CN)3(NO2)]PF6, a synthon for [Ru(bpy)(L3)NO2] complexes. AB - We report a high yield, two-step synthesis of fac-[Ru(bpy)(CH3CN)3NO2]PF6 from the known complex [(p-cym)Ru(bpy)Cl]PF6 (p-cym = eta(6)-p-cymene). [(p cym)Ru(bpy)NO2]PF6 is prepared by reacting [(p-cymene)Ru(bpy)Cl]PF6 with AgNO3/KNO2 or AgNO2. The 15NO2 analogue is prepared using K15NO2. Displacement of p-cymene from [(p-cym)Ru(bpy)NO2]PF6 by acetonitrile gives [Ru(bpy)(CH3CN)3NO2]PF6. The new complexes [(p-cym)Ru(bpy)NO2]PF6 and fac [Ru(bpy)(CH3CN)3NO2]PF6 have been fully characterized by 1H and 15N NMR, IR, elemental analysis, and single-crystal structure determination. Reaction of [Ru(bpy)(CH3CN)3NO2]PF6 with the appropriate ligands gives the new complexes [Ru(bpy)(Tp)NO2] (Tp = HB(pz)3-, pz = 1-pyrazolyl), [Ru(bpy)(Tpm)NO2]PF6 (Tpm = HC(pz)3), and the previously prepared [Ru(bpy)(trpy)NO2]PF6 (trpy = 2,2',6',2' ' terpyridine). Reaction of the nitro complexes with HPF6 gives the new nitrosyl complexes [Ru(bpy)TpNO][PF6]2 and [Ru(bpy)(Tpm)NO][PF6]3. All complexes were prepared with 15N-labeled nitro or nitrosyl groups. The nitro and nitrosyl complexes were characterized by 1H and 15N NMR and IR spectroscopy, elemental analysis, cyclic voltammetry, and single-crystal structure determination for [Ru(bpy)TpNO][PF6]2. For the nitro complexes, a linear correlation is observed between the nitro 15N NMR chemical shift and 1/nu(asym), where nu(asym) is the asymmetric stretching frequency of the nitro group. PMID- 17083259 TI - Electrochemistry of aluminum phthalocyanine: solvent and anion effects on UV visible spectra and reduction mechanisms. AB - The electrochemistry and UV-vis spectral properties of neutral and electroreduced Al(III) phthalocyanine, (Pc)AlCl, were characterized in four different nonaqueous solvents (THF, DMSO, DMF, and pyridine) containing tetra-n-butylammonium perchlorate, as well as in THF containing 0.4 M TBAP and the more strongly coordinating Cl-, F-, OH-, or CN- anions added to solution in the form of a tetra n-butylammonium salt. The initial phthalocyanine added to solution is represented as (Pc)AlCl, but the actual electroactive form of the compound varied as a function of both the solvent and type or number of bound anionic axial ligands. An uncharged (Pc)AlCl(THF) or (Pc)Al(CN)(THF) complex is present in THF solutions containing 0.4 M TBAP and excess Cl- or CN-, while transient mu-oxo dimers are spectroscopically observed upon addition of OH- or F- to (Pc)AlCl(THF) in THF followed by the ultimate formation of stable six-coordinate anionic species represented as [(Pc)Al(OH)2]- or [(Pc)AlF2]-. Each phthalocyanine undergoes three reversible one-electron additions at the conjugated Pc macrocycle within the negative potential limit of the solvent, and the UV-vis spectral changes obtained during the first two reductions were recorded in a thin-layer cell to evaluate the prevailing electron-transfer mechanisms. PMID- 17083260 TI - Modulation of the lowest metal-to-ligand charge-transfer state in [Ru(bpy)2(N N)]2+ systems by changing the N-N from hydrazone to azine: photophysical consequences. AB - Two Ru(II) complexes, [Ru(bpy)2L](ClO4)2 (1) and [Ru(bpy)2L'](BF4)2 (2), where bpy is 2,2'-bipyridine, L is diacetyl dihydrazone, and L' 1:2 is the condensate of L and acetone, are synthesized. From X-ray crystal structures, both are found to contain distorted octahedral RuN(6)(2+) cores. NMR spectra show that the cations in 1 and 2 possess a C2 axis in solution. They display the expected metal to-ligand charge transfer (1MLCT) band in the 400-500 nm region. Complex 1 is nonemissive at room temperature in solution as well as at 80 K. In contrast, complex 2 gives rise to an appreciable emission upon excitation at 440 nm. The room-temperature emission is centered at 730 nm (lambda(em)(max)) with a quantum yield (Phi(em)) of 0.002 and a lifetime (tau(em)) of 42 ns in an air-equilibrated methanol-ethanol solution. At 80 K, Phi(em) = 0.007 and tau(em) = 178 ns, with a lambda(em)(max) of 690 nm, which is close to the 0-0 transition, indicating an 3MLCT excited-state energy of 1.80 eV. The radiative rate constant (5 x 10(4) s( 1)) at room temperature and 80 K is almost temperature independent. From spectroelectrochemistry, it is found that bpy is easiest to reduce in 2 and that L is easiest in 1. The implications of this are that in 2 the lowest (3)MLCT state is localized on a bpy ligand and in 1 it is localized on L. Transient absorption results also support these assignments. As a consequence, even though 2 shows a fairly strong and long-lived emission from a Ru(II) --> bpy CT state, the Ru(II) --> L CT state in 1 shows no detectable emission even at 80 K. PMID- 17083261 TI - Better characterization of surface organometallic catalysts through resolution enhancement in proton solid state NMR spectra. AB - Delayed-acquisition methods, namely, echo and constant-time-acquisition approaches, allow a significant improvement in resolution in the proton solid state NMR spectra of surface organometallic catalysts such as [syn (SiO)Mo(=NAr)(=CH(t)Bu)(CH2(t)Bu)] and [(SiO)Re(C(t)Bu)(=CH(t)Bu)(CH2(t)Bu)] (syn/anti ratio = 1:1). This enables the observation of all of the proton resonances, which is not possible with the simple proton single-pulse technique under magic-angle spinning. For example, the methylene protons of the neopentyl ligands, buried in the large peak associated with all of the methyls in the 1H MAS spectrum, can easily be identified by recording a delayed-acquisition spectrum (resolution enhancement of a factor of 3 is obtained). Moreover, combining constant-time acquisition with heteronuclear carbon-proton correlation spectroscopy also improves the resolution of the 2D HETCOR spectra. PMID- 17083262 TI - Trapping of hemiquinone radicals at Mo and P sites by phosphide-bridged dimolybdenum species: chemistry of complexes [Mo2(eta5-C5H5)2(OC6H4OH)(mu PR2)(CO)4] and [Mo2(eta5-C5H5)2{mu-PR(OC6H4OH)}(CO)4]- (R = Cy, Ph). AB - The phosphide-bridged dimolybdenum complexes (H-DBU)[Mo2Cp2(mu-PR2)(CO)4] (R= Cy, Ph; DBU = 1,8-diazabicyclo[5.4.0.]undec-7-ene) react with p-benzoquinone to give the hemiquinone complexes [Mo(2)Cp2(OC6H4OH)(mu-PR2)(CO)4]. The latter experience facile homolytic cleavage of the corresponding Mo-O bonds and react readily at room temperature with HSPh or S2Ph2 to give the thiolate complexes [Mo2Cp2(mu PCy2)(mu-SPh)(CO)4] or [Mo2Cp2(mu-PR2)(mu-SPh)(CO)2]. In contrast, PRH-bridged substrates experience overall insertion of quinone into the P-H bond to give the anionic compounds (H-DBU)[Mo(2)Cp2{mu-PR(OC6H4OH)}(CO)4], which upon acidification yield the corresponding neutral hydrides. The cyclohexyl anion experiences rapid nucleophilic displacement of the hemiquinone group by different anions ER- (ER = OH, OMe, OC4H5, OPh, SPh) to give novel anionic compounds (H DBU)[Mo2Cp2{mu-PCy(ER)}(CO)4], which upon acidification yield the corresponding neutral hydrides. The structure of four of these hydride complexes [PPh(OC6H4OH), PCy(OH), PCy(OMe), and PCy(OPh) bridges] was determined by X-ray diffraction methods and confirmed the presence of cis and trans isomers in several of these complexes. In addition, it was found that the hydroxyphosphide anion [Mo2Cp2{mu PCy(OH)}(CO)4]- displays in solution an unprecedented tautomeric equilibrium with its hydride-oxophosphinidene isomer [Mo2Cp2(mu-H){mu-PCy(O)}(CO)4]-. PMID- 17083263 TI - Optimizing the pharmacologic treatment of hypertension: BP control and target organ protection. AB - Hypertension is a well documented risk factor for cardiovascular disease (CVD) and substantially contributes to the global burden of disease. Different drug options exist for combination therapy as part of an overall control of risk factors in order to decrease the absolute risk of CVD. Several guidelines in recent years have tried to set up recommendations to increase the proportion of subjects in acceptable BP control from high-risk groups. Some conventional drugs are still very important in modern hypertension treatment, e.g. low-dose thiazide diuretics. However, newer compounds have added to the list of useful agents to be used as monotherapy or in combination therapy for improved target organ protection, e.g. the calcium channel antagonists and ACE inhibitors. The role of beta-adrenoceptor antagonists (beta-blockers) has changed somewhat as a result of critical comments from recent meta-analyses. Currently, therefore, beta-blockers are recommended less often for primary prevention and monotherapy, but should still be used for secondary prevention, combination therapy, and for symptom relief. Finally, the new angiotensin II type 1 receptor antagonists (angiotensin receptor blockers [ARBs]) are still rather expensive, but have been increasingly documented in clinical trials for patients with essential hypertension. One controversial aspect is whether ARBs are better, worse, or equal to standard therapy with an ACE inhibitor for cardiovascular protection. BP remains poorly controlled in a large number of hypertensive patients and there is a greater need to control all relevant CVD risk factors in such patients. Therefore, different drugs are needed in order to be used in evidence-based synergistic and cost effective drug combinations. PMID- 17083264 TI - Thiazolidinediones in patients with diabetes mellitus and heart failure : implications of emerging data. AB - Individuals with diabetes mellitus have an increased risk of developing heart failure, usually as a consequence of coronary artery disease, although a specific diabetic cardiomyopathy, secondary to a microangiopathy, may also exist. The thiazolidinediones, a relatively new class of insulin-sensitizing agents used in the management of type 2 diabetes mellitus, have a number of complex metabolic actions on surrogate markers of atherogenesis, supported by the results of the recently published PROACTIVE (PROspective pioglitAzone Clinical Trial In macroVascular Events) trial. Unfortunately, the use of thiazolidinediones in individuals with diabetes mellitus and heart failure is limited because of a propensity to cause fluid retention. The underlying mechanisms of fluid retention have yet to be fully elucidated, but appear to be a dose-related class effect, exacerbated by combination therapy with insulin, and in some cases may be localized to peripheral edema. In parallel, echocardiographic studies show no significant effect of thiazolidinediones on cardiac structure or function. The design of epidemiologic studies describing an increased risk of developing heart failure in individuals with type 2 diabetes mellitus prescribed thiazolidinediones has been questioned, and a study of 'new users' of antihyperglycemic treatments found no increased risk of hospitalization for heart failure with thiazolidinedione therapy. There is also increasing evidence for the potential benefits of insulin sensitization in patients with diabetes mellitus and known heart failure, and a large observational study of over 16 000 patients with a principal discharge diagnosis of heart failure found a reduced mortality (hazard ratio [HR] 0.87; 95% CI 0.80, 0.94) in those prescribed thiazolidinediones. This benefit was offset by an increased risk of readmission with heart failure (HR 1.06; 95% CI 1.00, 1.09). Despite an increase in fluid related events, recent studies suggest that individuals with type 2 diabetes mellitus and heart failure (New York Heart Association grade I/II) can be treated with thiazolidinediones with appropriate monitoring and adjustment of heart failure therapies. These findings would suggest the need for large-scale, prospective trials to investigate the safety and potential benefits of thiazolidinedione use in patients with diabetes mellitus and heart failure. PMID- 17083265 TI - Therapeutic potential of thymosin-beta4 and its derivative N-acetyl-seryl aspartyl-lysyl-proline (Ac-SDKP) in cardiac healing after infarction. AB - Despite the numerous advances made in the prevention and treatment of cardiovascular diseases, there is a need for new strategies to repair and/or regenerate the myocardium after ischemia and infarction in order to prevent maladaptive remodeling and cardiac dysfunction. This article compiles and analyzes the available experimental data regarding the potential therapeutic effects of thymosin-beta4 and its derivative N-acetyl-seryl-aspartyl-lysyl proline (Ac-SDKP) in cardiac healing after myocardial infarction (MI) as well as discussing the possible mechanisms involved. The healing properties of thymosin beta4 have been described in different types of tissues, such as the skin and cornea, and more recently it has been shown that thymosin-beta4 facilitates cardiac repair after infarction by promoting cell migration and myocyte survival. Additionally, the tetrapeptide Ac-SDKP was reported to reduce left ventricular fibrosis in hypertensive rats, reverse fibrosis and inflammation in rats with MI, and stimulate both in vitro and in vivo angiogenesis. Ac-SDKP also reduced cardiac rupture rate in mice post-MI. Some of the effects of Ac-SDKP, such as the enhancement of angiogenesis and the decrease in inflammation and collagenase activity, are similar to those described for thymosin-beta4. Thus, it is possible that Ac-SDKP could be mediating some of the beneficial effects of its precursor. Although the experimental evidence is very promising, there are no data available from a clinical trial supporting the use of thymosin-beta(4) or Ac-SDKP as means of healing the myocardium after MI in patients. PMID- 17083266 TI - Medical treatment of patients with heart failure or left ventricular dysfunction undergoing percutaneous coronary intervention. AB - Management of ischemic patients with pre-existing or new-onset left ventricular (LV) dysfunction poses a special challenge in terms of the timing of percutaneous coronary intervention (PCI) and appropriate adjunctive medications to optimize outcome while minimizing risk. In a systematic fashion, this review attempts to provide a management scheme for patients with heart failure or LV dysfunction that present with stable angina, ST-segment elevation myocardial infarction, or unstable angina/non-ST-segment elevation myocardial infarction. By addressing therapeutic approaches to acute or decompensated heart failure and timing of coronary angiography based on severity of ischemia, we provide evidence-based recommendations for medications to initiate before, during, and following PCI. PMID- 17083267 TI - The ASCOT trial: clarifying the role of ACE inhibition in the reduction of cardiovascular events in patients with hypertension. AB - Hypertension remains one of the primary causes of preventable death in developed countries. Universally accepted clinical practice guidelines based on well designed trials are needed to ensure standardized treatment for these patients. It has been hypothesized that agents such as calcium channel antagonists and ACE inhibitors may be more effective than beta-adrenoceptor antagonist (beta-blocker) or diuretic-based regimens. However, until recently, there have been limited data on the efficacy of 'newer' antihypertensive regimens, particularly combination therapy, relative to standard therapeutic options. The ASCOT-BPLA (Anglo-Scandinavian Cardiac Outcomes Trial-Blood Pressure Lowering Arm), a prospective, randomized, controlled trial in >19 250 patients with hypertension, highlights the benefit of a combined amlodipine plus perindopril regimen over an atenolol plus diuretic-based combination in terms of significant reductions (p < 0.05) in all-cause mortality, total cardiovascular events and procedures, and fatal and nonfatal stroke. The ASCOT-BPLA trial underlines the cardiovascular benefit of ACE inhibitors, specifically perindopril, beyond that provided by BP reduction, and potentially reflects a mechanistic feature of the ACE inhibitors. It also suggests that the combination of amlodipine plus perindopril may provide broad-spectrum cardiovascular protection, as well as reduce the incidence of new onset diabetes mellitus and renal impairment, in addition to its efficacy in lowering BP. The ASCOT-BPLA trial provides evidence of the need to reconsider traditional prescribing recommendations in patients with hypertension, particularly conclusions reached in the US after the publication of the ALLHAT (Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial) study. The recently published National Institute for Health and Clinical Excellence (NICE) guidelines in the UK emphasize the efficacy of ACE inhibitors as first-line agents for hypertensive patients <55 years of age and recommend ACE inhibitor therapy at any stage of hypertension management. PMID- 17083268 TI - The protective effects of angiotensin II blockade with olmesartan medoxomil on resistance vessel remodeling (The VIOS study): rationale and baseline characteristics. AB - BACKGROUND: The VIOS (Vascular Improvement with Olmesartan medoxomil Study) study is a randomized, parallel study to determine the relative effects of suppressing the renin-angiotensin system (RAS) with the angiotensin receptor antagonist olmesartan medoxomil versus suppressing sympathetic drive with the beta adrenoceptor antagonist atenolol on remodeling of the subcutaneous small resistance vessel. Remodeling of small resistance vessels may be the earliest pathologic finding associated with hypertension. It may predate the onset of clinically apparent hypertension. METHODS: In this study, 100 patients with stage I hypertension are characterized at baseline before being treated for 1 year to obtain a goal BP of less than 140/90 mm Hg as defined by Joint National Committee (JNC)-7. Resistance vessel remodeling is determined using the gluteal fat biopsy technique in the hypertensive patients and a group of normotensive healthy volunteers. Additionally, efforts will be made to define whether noninvasive hemodynamic parameters, retinal vessel measurement changes, or biologic markers may predict and track the underlying vascular morphologic and physiologic changes induced by either regimen during the 12-month treatment period. RESULTS: The primary endpoint will be the degree of vascular remodeling as obtained from percutaneous biopsy of gluteal subcutaneous resistance vessels in each of two treatment arms compared with the normal volunteers. The design of the study and the pertinent baseline characteristics of these patients with uncomplicated essential hypertension are presented. CONCLUSION: The suppression of the RAS by the blockade of angiotensin II type 1 (AT(1)) receptors may demonstrate remodeling effects on the ubiquitous small resistance vessels similar to that seen in the myocardium and renal glomeruli, thus affording more complete end organ protection. PMID- 17083269 TI - The effect of moxonidine on endothelial dysfunction in metabolic syndrome. AB - BACKGROUND: Endothelial dysfunction has been reported in patients with type 2 diabetes mellitus and even in healthy obese individuals with a normal metabolic profile. Sympathetic activity commonly is increased in obese hypertensive patients, and moxonidine is effective in lowering BP and improving insulin sensitivity. OBJECTIVE: To evaluate the effect of moxonidine on endothelial dysfunction in patients with metabolic syndrome. METHODS: Twenty-six patients with mild hypertension were treated with moxonidine and a hypocaloric diet for 3 months, while a second normotensive group (n = 26) were followed-up with calorie restriction alone. Anthropometric (body mass index, waist and hip circumferences, and waist-to-hip ratio) and metabolic features (fasting plasma glucose and insulin, aminotransferases, gamma-glutamyl transpeptidase, triglycerides, and cholesterol levels) and flow-mediated dilatation (FMD) were evaluated. Insulin resistance was calculated by using the homeostasis model assessment formula. Insulin sensitivity was calculated according to the quantitative insulin sensitivity check index (QUICKI). RESULTS: SBP and DBP (both p < 0.001) and waist circumference (p = 0.02) were higher, and QUICKI (p = 0.043) and FMD (p = 0.01) were lower in the hypertensive group at baseline. After 3 months, nearly all the study parameters improved in both treatment groups. The decrease in BP, increase in FMD, and improvements in metabolic and anthropometric parameters were significantly greater in the moxonidine-treated group than in those treated with diet alone. CONCLUSION: Moxonidine is proposed as a valuable option for treating mild-to-moderate hypertension in obese and insulin-resistant patients with metabolic syndrome as it appears to improve endothelial dysfunction in these patients. PMID- 17083270 TI - Evaluation of lipid-lowering therapy and cholesterol goal attainment in Finland: the National FINRISK Study. AB - BACKGROUND: European and US study findings show that patients in primary care and specialty care are not adequately treated for cholesterol reduction. OBJECTIVE: To evaluate lipid-lowering treatment in Finland, estimate the proportions of subjects who are achieving cholesterol goals, and assess the influence of determinants on goal attainment. METHODS: Subgroup analysis of the FINRISK study, a national study of cardiovascular disease risk factors in Finland. Study participants, the subgroup of patients on lipid-lowering therapy from FINRISK, completed a postal self-administered questionnaire on health/health behavioral factors. Serum total cholesterol (TC) and other clinical variables were measured using a standardized protocol. Ten-year coronary risk was computed using Framingham risk equations. The influence of certain factors on goal attainment was determined by logistic regression analysis. The main outcome measure was the proportion of subjects who were receiving lipid-lowering therapy and achieved a TC goal of <5 mmol/L (<194 mg/dL). RESULTS: Among 9581 respondents, 622 subjects were on lipid-lowering therapy. Of these, 68 subjects were excluded because of missing data on TC and/or the type/dose of therapy. Among the 554 subjects included, 210 (38%) were secondary-prevention patients and 51% had 10-year coronary risk >/=20%. Approximately two-thirds of subjects were prescribed simvastatin (42%) or atorvastatin (26%), and about half (51%) were prescribed low equipotency HMG-CoA reductase inhibitors (statins). There was no difference in equipotent doses of statins prescribed for primary and secondary prevention. About half (54%) of subjects did not attain their cholesterol goal (TC <5.0 mmol/L [194 mg/dL]). Subjects with coronary heart disease (odds ratio [OR] 3.00; 95% CI 2.07, 4.35) and patients prescribed medium-to-high equipotent statins (OR 1.93; 95% CI 1.35, 2.76) were more likely to achieve cholesterol goals, whereas postmenopausal women (OR 0.61; 95% CI 0.42, 0.88) were less likely than men to achieve cholesterol goals. CONCLUSION: Most (92%) patients receiving lipid lowering therapy in a Finnish population were managed on statin monotherapy and approximately half of the patients did not achieve their recommended cholesterol goals. More effective and safe therapies are needed to enhance cholesterol goal attainment. These treatments might include regimens that act on two or more pharmacologic pathways. PMID- 17083271 TI - Spotlight on ranolazine in chronic stable angina pectoris. AB - Ranolazine (Ranexa), a piperazine derivative, is a new antianginal agent approved for the treatment of chronic stable angina pectoris for use as combination therapy when angina is not adequately controlled with other antianginal agents. While the exact mechanism of action of ranolazine is not known, its antianginal and anti-ischemic effects do not appear to depend upon changes in BP or heart rate. An extended-release (ER) oral formulation of ranolazine has been developed to facilitate twice-daily administration whilst maintaining therapeutically effective plasma concentrations. In patients with chronic stable angina, ranolazine ER monotherapy was shown to improve exercise duration at trough plasma drug concentration in a dose-dependent manner compared with placebo. The drug was effective as adjunctive therapy in patients with chronic stable angina whose condition was not controlled adequately with conventional antianginal therapy. In randomized clinical trials, ranolazine ER was well tolerated, with no overt effects on cardiovascular hemodynamics or conduction, apart from a modest increase in corrected QT interval (but no torsades de pointes). Importantly, the efficacy and tolerability of ranolazine ER were not affected by old age and co morbid conditions (heart failure or diabetes mellitus). Comparative trials of ranolazine ER with other antianginal agents and trials examining its effects on long-term morbidity and mortality in patients with ischemic heart disease are required to determine with greater certainty the place of the drug in current antianginal therapy. Nevertheless, ranolazine ER may well prove to be a useful alternative and adjunct to conventional hemodynamic antianginal therapy in the treatment of chronic stable angina. PMID- 17083272 TI - The evolution of two-component systems in bacteria reveals different strategies for niche adaptation. AB - Two-component systems including histidine protein kinases represent the primary signal transduction paradigm in prokaryotic organisms. To understand how these systems adapt to allow organisms to detect niche-specific signals, we analyzed the phylogenetic distribution of nearly 5,000 histidine protein kinases from 207 sequenced prokaryotic genomes. We found that many genomes carry a large repertoire of recently evolved signaling genes, which may reflect selective pressure to adapt to new environmental conditions. Both lineage-specific gene family expansion and horizontal gene transfer play major roles in the introduction of new histidine kinases into genomes; however, there are differences in how these two evolutionary forces act. Genes imported via horizontal transfer are more likely to retain their original functionality as inferred from a similar complement of signaling domains, while gene family expansion accompanied by domain shuffling appears to be a major source of novel genetic diversity. Family expansion is the dominant source of new histidine kinase genes in the genomes most enriched in signaling proteins, and detailed analysis reveals that divergence in domain structure and changes in expression patterns are hallmarks of recent expansions. Finally, while these two modes of gene acquisition are widespread across bacterial taxa, there are clear species specific preferences for which mode is used. PMID- 17083273 TI - The evolution of word composition in metazoan promoter sequence. AB - The field of molecular evolution provides many examples of the principle that molecular differences between species contain information about evolutionary history. One surprising case can be found in the frequency of short words in DNA: more closely related species have more similar word compositions. Interest in this has often focused on its utility in deducing phylogenetic relationships. However, it is also of interest because of the opportunity it provides for studying the evolution of genome function. Word-frequency differences between species change too slowly to be purely the result of random mutational drift. Rather, their slow pattern of change reflects the direct or indirect action of purifying selection and the presence of functional constraints. Many such constraints are likely to exist, and an important challenge is to distinguish them. Here we develop a method to do so by isolating the effects acting at different word sizes. We apply our method to 2-, 4-, and 8-base-pair (bp) words across several classes of noncoding sequence. Our major result is that similarities in 8-bp word frequencies scale with evolutionary time for regions immediately upstream of genes. This association is present although weaker in intronic sequence, but cannot be detected in intergenic sequence using our method. In contrast, 2-bp and 4-bp word frequencies scale with time in all classes of noncoding sequence. These results suggest that different genomic processes are involved at different word sizes. The pattern in 2-bp and 4-bp words may be due to evolutionary changes in processes such as DNA replication and repair, as has been suggested before. The pattern in 8-bp words may reflect evolutionary changes in gene-regulatory machinery, such as changes in the frequencies of transcription-factor binding sites, or in the affinity of transcription factors for particular sequences. PMID- 17083274 TI - Falstatin, a cysteine protease inhibitor of Plasmodium falciparum, facilitates erythrocyte invasion. AB - Erythrocytic malaria parasites utilize proteases for a number of cellular processes, including hydrolysis of hemoglobin, rupture of erythrocytes by mature schizonts, and subsequent invasion of erythrocytes by free merozoites. However, mechanisms used by malaria parasites to control protease activity have not been established. We report here the identification of an endogenous cysteine protease inhibitor of Plasmodium falciparum, falstatin, based on modest homology with the Trypanosoma cruzi cysteine protease inhibitor chagasin. Falstatin, expressed in Escherichia coli, was a potent reversible inhibitor of the P. falciparum cysteine proteases falcipain-2 and falcipain-3, as well as other parasite- and nonparasite derived cysteine proteases, but it was a relatively weak inhibitor of the P. falciparum cysteine proteases falcipain-1 and dipeptidyl aminopeptidase 1. Falstatin is present in schizonts, merozoites, and rings, but not in trophozoites, the stage at which the cysteine protease activity of P. falciparum is maximal. Falstatin localizes to the periphery of rings and early schizonts, is diffusely expressed in late schizonts and merozoites, and is released upon the rupture of mature schizonts. Treatment of late schizionts with antibodies that blocked the inhibitory activity of falstatin against native and recombinant falcipain-2 and falcipain-3 dose-dependently decreased the subsequent invasion of erythrocytes by merozoites. These results suggest that P. falciparum requires expression of falstatin to limit proteolysis by certain host or parasite cysteine proteases during erythrocyte invasion. This mechanism of regulation of proteolysis suggests new strategies for the development of antimalarial agents that specifically disrupt erythrocyte invasion. PMID- 17083275 TI - Selection acts on DNA secondary structures to decrease transcriptional mutagenesis. AB - Single-stranded DNA is more subject to mutation than double stranded. During transcription, DNA is transiently single stranded and therefore subject to higher mutagenesis. However, if local intra-strand secondary structures are formed, some bases will be paired and therefore less sensitive to mutation than unpaired bases. Using complete genome sequences of Escherichia coli, we show that local intra-strand secondary structures can, as a consequence, be used to define an index of transcription-driven mutability. At gene level, we show that natural selection has favoured a reduced transcription-driven mutagenesis via the higher than expected frequency of occurrence of intra-strand secondary structures. Such selection is stronger in highly expressed genes and suggests a sequence-dependent way to control mutation rates and a novel form of selection affecting the evolution of synonymous mutations. PMID- 17083276 TI - CAF-1 is essential for heterochromatin organization in pluripotent embryonic cells. AB - During mammalian development, chromatin dynamics and epigenetic marking are important for genome reprogramming. Recent data suggest an important role for the chromatin assembly machinery in this process. To analyze the role of chromatin assembly factor 1 (CAF-1) during pre-implantation development, we generated a mouse line carrying a targeted mutation in the gene encoding its large subunit, p150CAF-1. Loss of p150CAF-1 in homozygous mutants leads to developmental arrest at the 16-cell stage. Absence of p150CAF-1 in these embryos results in severe alterations in the nuclear organization of constitutive heterochromatin. We provide evidence that in wild-type embryos, heterochromatin domains are extensively reorganized between the two-cell and blastocyst stages. In p150CAF-1 mutant 16-cell stage embryos, the altered organization of heterochromatin displays similarities to the structure of heterochromatin in two- to four-cell stage wild-type embryos, suggesting that CAF-1 is required for the maturation of heterochromatin during preimplantation development. In embryonic stem cells, depletion of p150CAF-1 using RNA interference results in the mislocalization, loss of clustering, and decondensation of pericentric heterochromatin domains. Furthermore, loss of CAF-1 in these cells results in the alteration of epigenetic histone methylation marks at the level of pericentric heterochromatin. These alterations of heterochromatin are not found in p150CAF-1-depleted mouse embryonic fibroblasts, which are cells that are already lineage committed, suggesting that CAF-1 is specifically required for heterochromatin organization in pluripotent embryonic cells. Our findings underline the role of the chromatin assembly machinery in controlling the spatial organization and epigenetic marking of the genome in early embryos and embryonic stem cells. PMID- 17083277 TI - Recombination hotspots flank the Cryptococcus mating-type locus: implications for the evolution of a fungal sex chromosome. AB - Recombination increases dramatically during meiosis to promote genetic exchange and generate recombinant progeny. Interestingly, meiotic recombination is unevenly distributed throughout genomes, and, as a consequence, genetic and physical map distances do not have a simple linear relationship. Recombination hotspots and coldspots have been described in many organisms and often reflect global features of chromosome structure. In particular, recombination frequencies are often distorted within or outside sex-determining regions of the genome. Here, we report that recombination is elevated adjacent to the mating-type locus (MAT) in the pathogenic basidiomycete Cryptococcus neoformans. Among fungi, C. neoformans has an unusually large MAT locus, and recombination is suppressed between the two >100-kilobase mating-type specific alleles. When genetic markers were introduced at defined physical distances from MAT, we found the meiotic recombination frequency to be approximately 20% between MAT and a flanking marker at 5, 10, 50, or 100 kilobases from the right border. As a result, the physical/genetic map ratio in the regions adjacent to MAT is distorted approximately 10- to 50-fold compared to the genome-wide average. Moreover, recombination frequently occurred on both sides of MAT and negative interference between crossovers was observed. MAT heterozygosity was not required for enhanced recombination, implying that this process is not due to a physical distortion from the two non-paired alleles and could also occur during same-sex mating. Sequence analysis revealed a correlation between high G + C content and these hotspot regions. We hypothesize that the presence of recombinational activators may have driven several key events during the assembly and reshaping of the MAT locus and may have played similar roles in the origins of both metabolic and biosynthetic gene clusters. Our findings suggest that during meiosis the MAT locus may be exchanged onto different genetic backgrounds and therefore have broad evolutionary implications with respect to mating-type switching in both model and pathogenic yeasts. PMID- 17083278 TI - Medical students' assessments of skill development in rural primary care clinics. AB - INTRODUCTION: This study assessed the impact of a rural primary care preceptorship on medical students' self-perceived ability to provide acute, chronic, and preventive care, to perform procedures, to communicate with patients, and to understand the community and healthcare system. METHODS: Students were surveyed about their self-assessed skills on 11 major components (97 items) immediately before and after a 16 week preceptorship in a rural primary care clinic. Responses were analyzed for 96 medical students using a paired comparisons t-test and univariate statistics. RESULTS: Students' skills significantly increased on all components and items. The skills most highly assessed post-preceptorship were those skills related to the management of chronic problems, the provision of patient education and health maintenance, and the ability to handle undifferentiated and acute problems. Among the 11 components assessed, students ranked their skills in performing procedures the lowest. The largest cumulative gain in skills was in the areas of understanding health systems and the community. CONCLUSIONS: This study provides a unique opportunity to look at skill development before and after a rural clerkship. From the student's perspective, the 16 week preceptorship appears to be of significant educational benefit. Future studies need to examine other measures of performance and outcomes of training in rural primary care settings. PMID- 17083279 TI - Tobacco use among high school students in a remote district of Arua, Uganda. AB - INTRODUCTION: Tobacco smoking is a risk factor for several non-communicable public health problems including cancer, ischaemic heart disease and chronic obstructive airways disease. The prevalence of smoking among adolescents and the associated environment deserve attention. METHODS: A cross-sectional descriptive study was carried out in 2001 to determine the prevalence of tobacco smoking, exposure to advertisements, environmental tobacco smoke exposure, deterrents from smoking and perception about smoking among high school students in a remote district of Arua, north-western Uganda. RESULTS: In total 1528 high school students participated in the study of which 21.9% were current smokers and 33.1% had ever used tobacco products. When the data were stratified according to sex, 81/452 (17.9%) females and 337/871 (38.7%) males had ever smoked (p <0.05). With regard to current smoking, defined as having smoked at least once in the past 30 days, 12.2% of females (55/452) and 25.5% males were current smokers. Approximately one-tenth perceived themselves as likely to initiate smoking within the next 12 months. Just under half (47.3%) were exposed to environmental (passive) smoking in the home and 19.4% of current smokers usually smoked at home. Approximately 60% of smokers had obtained tobacco from grocery stores and they had never been prevented because of their age. Media exposure to tobacco advertisements was high. CONCLUSION: Many young people in Arua, Uganda, were current smokers and exposed to environments that seemed to facilitate uptake of tobacco smoking and other tobacco use. This could be explained in part, by the fact that the district relies heavily on tobacco farming and exposure to facilitating environments is common. A concerted public health response is urgently required that will effectively alter the home and societal environment so as to discourage uptake of tobacco use by young people. PMID- 17083280 TI - Visual search: the role of peripheral information measured using gaze-contingent displays. AB - Two of the factors limiting progress in understanding the mechanisms of visual search are the difficulty of controlling and manipulating the retinal stimulus when the eyes are free to move and the lack of an ideal observer theory for fixation selection during search. Recently, we developed a method to precisely control retinal stimulation with gaze-contingent displays (J. S. Perry & W. S. Geisler, 2002), and we derived a theory of optimal eye movements in visual search (J. Najemnik & W. S. Geisler, 2005). Here, we report a parametric study of visual search for sine-wave targets added to spatial noise backgrounds that have spectral characteristics similar to natural images (the amplitude spectrum of the noise falls inversely with spatial frequency). Search time, search accuracy, and eye fixations were measured as a function of target spatial frequency, 1/f noise contrast, and the resolution falloff of the display from the point of fixation. The results are systematic and similar for the two observers. We find that many aspects of search performance and eye movement pattern are similar to those of an ideal searcher that has the same falloff in resolution with retinal eccentricity as the human visual system. PMID- 17083281 TI - Humans can perceive heading without visual path information. AB - It has previously been reported that humans can determine their direction of 3D translation (heading) from the 2D velocity field of retinal motion experienced during self-motion through a rigid environment, as is done by current computational models of visual heading estimation from optic flow. However, these claims were supported by studies that used stimuli that contained low rotational flow rates and/or additional visual cues beyond the velocity field or a task in which observers were asked to indicate their future trajectory of self-motion (path). Thus, previous conclusions about heading estimation have been confounded by the presence of other visual factors beyond the velocity field, by the use of a path-estimation task, or both. In particular, path estimation involves an exocentric computation with respect to an environmental reference, whereas heading estimation is an egocentric computation with respect to one's line of sight. Here, we use a heading-adjustment task to demonstrate that humans can precisely estimate their heading from the velocity field, independent of visual information about path, displacement, layout, or acceleration, with accuracy robust to rotation rates at least as high as 20 deg/s. Our findings show that instantaneous velocity-field information about heading is directly available for the visual control of locomotion and steering. PMID- 17083282 TI - Perception of surface slant from oriented textures. AB - When a surface covered with a regular texture is viewed in perspective, the projected texture provides a number of cues to 3D surface orientation. For oriented textures, one cue is perspective convergence: symmetry lines that are parallel along the surface project to lines that vary systematically in orientation. We investigated the contribution of perspective convergence to perception of 3D slant and tested whether slant from convergence depends on oriented spectral components. Subjects judged the sign of slant about a vertical axis of rotation. Textures were composed of filled circles in three spatial arrangements: a hex grid with symmetry lines at 0 and +/-60 deg relative to the tilt direction (aligned condition), a hex grid with symmetry lines at 90 and +/ 30 deg (perpendicular condition), and random arrangements with similar average spacing (isotropic condition). The two hex grid textures differed in the amount of spectral energy present in the tilt direction (horizontal) but were otherwise closely matched. Slant discrimination thresholds for monocular stimuli were higher for isotropic textures than for either of the two hex grid textures and were higher for the perpendicular texture than for the aligned texture. In a second experiment, we measured the weight given to texture relative to binocular slant information for cue conflict stimuli (+/-5 deg). Weights were found to agree with individual subjects' monocular thresholds, in accordance with optimal estimation theory. We conclude that the visual system uses perspective convergence to perceive slant and that effective use of convergence requires the presence of spectral components aligned with the tilt direction. PMID- 17083283 TI - The role of memory in guiding attention during natural vision. AB - What is the time frame in which perceptual memory guides attention? Current estimates range from a few hundred milliseconds to several seconds, minutes, or even days. Here, we answer this question by establishing the time course of attentional selection in realistic viewing conditions. First, we transformed continuous video clips into MTV-style video clips by stringing together continuous clip segments using abrupt transitions (jump cuts). We then asked participants to visually explore either continuous or MTV-style clips and recorded their saccades as objective behavioral indicators of attentional selections. The utilization of perceptual memory was estimated across viewing conditions and over time by quantifying the agreement between human attentional selections and predictions made by a neurally grounded computational model. In the critical condition, jump cuts led to sharp declines in the impact of perceptual memory on attentional selection, followed by monotonic increases in memory utilization across seven consecutive saccades and 2.5 s. These results demonstrate that perceptual memory traces play an important role in guiding attention across several saccades during natural vision. We propose novel hypotheses and experiments using hybrid natural-artificial stimuli to further elucidate neurocomputational mechanisms of attentional selection. PMID- 17083284 TI - The mechanisms of collinear integration. AB - Low-contrast visual contour fragments are easier to detect when presented in the context of nearby collinear contour elements (U. Polat & D. Sagi, 1993). The spatial and temporal determinants of this collinear facilitation have been studied extensively (J. R. Cass & B. Spehar, 2005; Y. Tanaka & D. Sagi, 1998; C. B. Williams & R. F. Hess, 1998), although considerable debate surrounds the neural mechanisms underlying it. Our study examines this question using a novel stimulus, whereby the flanking "contour" elements are rotated around their own axis. By measuring contrast detection thresholds to a brief foveal target presented at various phases of flanker rotation, we find peak facilitation after flankers have rotated beyond their collinear phase. This optimal facilitative delay increases monotonically as a function of target-flanker separation, yielding estimates of cortical propagation of 0.1 m/s, a value highly consistent with the dynamics of long-range horizontal interactions observed within primary visual cortex (V1). A curious new finding is also observed: Facilitative peaks also occur when the target flash precedes flanker collinearity by 20-80 ms, a range consistent with contrast-dependent cortical onset latencies. Together, these data suggest that collinear facilitation involves two separate mechanisms, each possessing distinct dynamics: (i) slowly propagating horizontal interactions within V1 and (ii) a faster integrative mechanism, possibly driven by synchronous collinear cortical onset. PMID- 17083285 TI - Vernier acuity of illusory contours defined by motion. AB - We present here a series of experiments exploring a special class of visual completion that is strictly tied to the perception of apparent motion. The stimuli consist of sparse random-dot arrays, in which dots remain in place. Changes of luminance or color of the dots at leading and trailing edges of an apparently moving region are integrated over space and time to produce the perception of well-defined contours, shapes, and color. We test how Vernier acuity of apparent motion-defined illusory bars depends on speed, density, and stimulus configurations. We found that higher speed of apparent motion reduces the Vernier acuity thresholds. These thresholds also decrease with increasing density of dots, whose luminance changes provide the apparent motion signal required for the perception of illusory contours. In subsequent experiments, we showed that luminance-defined flankers could seamlessly integrate with and improve the perception of apparent motion-defined contours, reducing their Vernier thresholds. PMID- 17083286 TI - The accuracy and reliability of perceived depth from linear perspective as a function of image size. AB - We investigated the ability to use linear perspective to perceive depth from monocular images. Specifically, we focused on the information provided by convergence of parallel lines in an image due to perspective projection. Our stimuli were trapezoid-shaped projected contours, which appear as rectangles slanted in depth. If converging edges of a contour are assumed to be parallel edges of a 3D object, then it is possible in principle to recover its 3D orientation and relative dimensions. This 3D interpretation depends on projected size; hence, if an image contour were scaled, accurate use of perspective predicts changes in perceived slant and shape. We tested this prediction and measured the accuracy and precision with which observers can judge depth from perspective alone. Observers viewed monocular images of slanted rectangles and judged whether the rectangles appeared longer versus wider than a square. The projected contours had varying widths (7, 14, or 21 deg) and side angles (7 or 25 deg), and heights were varied by a staircase procedure to compute a point of subjective equality and 75% threshold for each condition. Observers were able to reliably judge aspect ratios from the monocular images: Weber fractions were 6-9% for the largest rectangles, increasing to as high as 17% for small rectangles with high simulated slant. Overall, the contours judged to be squares were taller than the projections of actual squares, consistent with perceptual underestimation of depth. Judgments were modulated by image size in the direction expected from perspective geometry, but the effect of size was only about 20-30% of what was predicted. We simulated the performance of a Bayesian ideal observer that integrated perspective information with an a priori bias toward compression of depth and which was able to qualitatively model the pattern of results. PMID- 17083287 TI - Transient covert attention and the perceived rate of flicker. AB - Transient covert attention affects basic visual dimensions such as contrast sensitivity, spatial resolution, and temporal resolution. Two recent studies provide evidence of corresponding phenomenological changes: The increase in contrast sensitivity and spatial resolution at the attended location is associated with increased apparent contrast (M. Carrasco, S. Ling, & S. Read, 2004) and apparent spatial frequency (J. Gobell & M. Carrasco, 2005). Here, we assessed a phenomenological correlate of attention for temporal vision, asking whether and how transient attention affects perceived flicker rate. We employed a psychophysical method developed to assess changes in appearance by manipulating transient attention via uninformative spatial cues. In each trial, two suprathreshold Gabor stimuli, appearing briefly to the left and right of fixation, were counterphase modulated at either the same or different temporal frequencies. To assess appearance, we asked observers to perform an orientation discrimination task contingent on perceived flicker rate: "What was the orientation of the Gabor that flickered faster?" Results indicated that perceived flicker rate increased at the cued location. A control experiment, in which observers reported the orientation of the Gabor that flickered slower, ruled out a cue bias explanation. We conclude that transient attention increases perceived flicker rate. PMID- 17083288 TI - Two cases requiring external reinforcement in perceptual learning. AB - The role of external reinforcement is an issue of much debate and uncertainty in perceptual learning research. Although it is commonly acknowledged that external reinforcement, such as performance feedback, can aid in perceptual learning (M. H. Herzog & M. Fahle, 1997), there are many examples in which it is not required (K. Ball & R. Sekuler, 1987; M. Fahle, S. Edelman, & T. Poggio, 1995; A. Karni & D. Sagi, 1991; S. P. McKee & G. Westheimer, 1978; L. P. Shiu & H. Pashler, 1992). Additionally, learning without external reinforcement can occur even for stimuli that are irrelevant to the subject's task (A. R. Seitz & T. Watanabe, 2003). It has been thus hypothesized that internal reinforcement can serve a similar role as external reinforcement in learning (M. H. Herzog & M. Fahle, 1998; A. Seitz & T. Watanabe, 2005). This idea suggests that perceptual learning should occur in the absence of external reinforcement provided that easy exemplars are utilized as a basis for the subject to generate internal reinforcement. Here, we report results from two studies that show that this is not always the case. In the first study, subjects participated in two sessions of a motion direction discrimination task with low-contrast dots moving in directions separated by 90 degrees. In the second study, subjects participated in 12 orientation-discrimination sessions using oriented bars (oriented either 70 degrees or 110 degrees) that were masked by spatial noise. Trials of different signal levels (yielding psychometric functions ranging from chance to ceiling) were randomly interleaved. In both studies, subjects experiencing external reinforcement showed significant learning, whereas subjects receiving no external reinforcement failed to show learning. We conclude that while internal reinforcement is an important learning signal, the presence of easy exemplars is not sufficient to generate reinforcement signals. PMID- 17083289 TI - Perceptual asynchrony between color and motion with a single direction change. AB - When a stimulus repeatedly and rapidly changes color (e.g., between red and green) and motion direction (e.g., upwards and downwards) with the same frequency, it was found that observers were most likely to pair colors and motion directions when the direction changes lead the color changes by approximately 80 ms. This is the color-motion asynchrony illusion. According to the differential processing time model, the illusion is explained because the neural activity leading to the perceptual experience of motion requires more time than that of color. Alternatively, the time marker model attributes the misbinding to a failure in matching different sorts of changes at rapid alternations. Here, running counter to the time marker model, we demonstrate that the illusion can arise with a single direction change. Using this simplified version of the illusion we also show that, although some form of visual masking takes place between colors, the measured asynchrony genuinely reflects processing time differences. PMID- 17083290 TI - Role of focal attention on latencies and trajectories of visually guided manual pointing. AB - Previous studies have shown that an odd-colored target among uniformly colored distractors can be rapidly detected and localized using broadly distributed attention over an entire display. In the current study, we show that such a broadly distributed attentional allocation is not sufficient for seemingly effortless goal-directed manual pointing. Latencies and movement durations of manual pointing in odd-colored search tasks become shorter and curved trajectories decreased as the number of distractors increase or target color repetitions increases. Because these manipulations have been shown to facilitate the deployment of narrowly focused attention to a target but not for distributed attention, this adds further support to the view that focal attention is necessary for goal-directed action. In addition, the presence of highly curved movement trajectories, directed first to a distractor then to the target reflects ongoing changes in focal attentional deployment and target selection. PMID- 17083291 TI - Nonlinearities in color coding: compensating color appearance for the eye's spectral sensitivity. AB - Most wavelengths change hue when mixed with white light. These changes, known as the Abney effect, have been extensively studied to characterize nonlinearities in the neural coding of color, but their potential function remains obscure. We measured the Abney effect in a new way--by varying the bandwidth of the spectrum rather than mixing with white--and this leads to a new interpretation of the role of nonlinear responses in color appearance. Because of the eye's limited spectral sensitivity, increasing the bandwidth of a spectrum changes the relative responses in the three classes of cone receptor and thus would change hue if the percept were tied to a fixed cone ratio. However, we found that hue is largely independent of bandwidth and thus constant for a constant peak wavelength for stimuli with Gaussian spectra. This suggests that color appearance is compensated for the eye's spectral filtering, and that this compensation embodies specific perceptual inferences about how natural spectra vary. When a wavelength is instead diluted with white light--which does not bias the cone ratios--then the same compensation predicts changes in hue because the "right" response is made to the "wrong" stimulus. This model generates constant hue loci that are qualitatively consistent with measures of the Abney effect and provides a novel functional account of such effects in color appearance, in which postreceptoral responses are adjusted so that constant hue percepts are tied to consistent physical properties of the environment rather than consistent physiological properties such as the cone ratios. PMID- 17083292 TI - Global and local processing in Williams syndrome, autism, and Down syndrome: perception, attention, and construction. AB - Global and local processing was studied in Williams Syndrome (WS), autism (AS), and Down Syndrome (DS) using perception, attention, and construction tasks. Past research has suggested an abnormal bias toward global processing in DS and, in contrast, an abnormal local bias in both WS and AS. Until now, no study has investigated whether the local processing bias in WS and AS has a different or similar underlying cause. Findings here suggest a common underlying mechanism, namely a bias in attention toward local processing. Results also indicate a global bias in attention in DS. This study finds no evidence to support predictions of the hierarchical deficit theory (Mottron & Belleville, 1993) as an explanation of hierarchical processing deficits in AS or DS, but does find support for hierarchical deficit theory in a subset of WS individuals. This study finds evidence of cognitive heterogeneity in WS, consistent with Porter and Coltheart (2005). PMID- 17083293 TI - Does attentional cueing affect dichotic listening performance in children? AB - This study addresses the effects of verbal versus nonverbal (tone) shifts of attention on dichotic listening (DL) performance with children. Theoretically, a tonal cue may be more effective in increasing attention than a verbal cue following instruction. The inconsistency of studies reporting substantial effects of attention on ear asymmetries in children with or without learning disabilities (LDs) may be due to a developmental difference in their ability to use verbal or tone cues to select stimuli for recall. Participants included 30 right-handed children (15 control, 15 with LDs) with a mean age of 10.8 years. Each participant received 60 trials of a monaural tone cue task, 60 trials of a binaural verbal cue task, and 60 trials of a monaural verbal cue task, to direct attention to either the left or right ear before the presentation of consonant vowel syllable pairs in a DL task. A factorial design analysis of variance yielded a significant right-ear advantage for both groups. More important, the Group x Task interaction was found to be significant, indicating that group performance on ear scores was dependent on type of cueing condition. Whereas all 3 cue conditions were effective in orienting attention for control participants, larger shifts were apparent under both binaural and monaural verbal instructional cue conditions. In contrast, participants with LD showed larger shifts of attention under the tonal cue condition. These results show that control participants have greater ability to focus attention with the use of a verbal cue, whereas participants with LD show greater ability to orient attention with the use of a tone cue in reducing error rates in DL performance. PMID- 17083294 TI - Memory abilities in children with subtypes of dyscalculia. AB - This study examines (a) mathematical skills of 2 subgroups of children with developmental dyscalculia (DD)--1 group with DD only and a second group with DD plus reading disorders (RDD)-and (b) analyzes the memory skills of both groups of children. Fifty 11- and 12-year-old children were selected from public schools in Guadalajara, Mexico. Seventeen children had DD only, 13 had RDD, and 20 were normal controls. Testing included 10 calculation and 6 memory subtests taken from the Evaluacion Neuropsicologica Infantil (Matute, Rosselli, Ardila, and Ostrosky, in press). Results indicated that children with DD and children with RDD show a similar pattern of mathematical impairment. Both subgroups had significantly lower scores than the control group in working memory tasks. In addition, the RDD group had significantly lower scores than the control group in visual learning and semantic memory. Although the RDD group scored lower than the DD group in most memory tests, this difference did not reach significance. Working memory tests (digits backwards and sentence repetition) appeared to be the best predictors of mathematical test scores and may represent a major cognitive defect in children with specific defects in mathematics. PMID- 17083295 TI - Delayed-execute prospective memory performance: the effects of age and working memory. AB - This study follows the novel delayed-execute prospective memory paradigm, which involves briefly delaying the execution of an intended action, a task that has been shown to produce substantial age effects. During the ongoing task, sentences were presented, and participants had to answer reading-comprehension questions and general knowledge questions. In the prospective memory task, the participant was to press a key after the presentation of a specific cue in the sentences-but not before a subsequent phase of the ongoing task was reached. In contrast to previous studies using older participants taken from very broadly defined age ranges, this study examines development of delayed-execute prospective memory more precisely by examining a total of 4 age groups: a younger age group (age range = 22-31; n = 27), a young-old age group (age range = 60-69; n = 34), a middle-old age group (age range = 70-79; n = 31), and an old-old age group (age range = 80-91; n = 35). This study investigates the dependence of (age-related) delayed-execute prospective memory performance on working memory capacity by disrupting the phonological loop during the delay period as well as its dependence on neuropsychological processes such as inhibitory control and processing speed. The results show that (a) delayed-execute prospective memory particularly declines within the group of older participants, (b) delayed-execute prospective memory is diminished when working memory load is high during the delay period, and (c) age-related performance in delayed-execute prospective memory may be mediated by inhibitory control. The findings are discussed in the context of the frontal lobe hypothesis of cognitive aging. PMID- 17083296 TI - Inhibitory control in children with phenylketonuria. AB - Past studies have reported impairments in children with early-treated phenylketonuria (PKU) in executive abilities such as strategic processing and working memory. Findings have been inconsistent in terms of the integrity of inhibitory control, another executive ability. This study administered 4 inhibitory tasks (flanker, Stroop, go/no-go, antisaccade) to 26 children with PKU and 25 typically developing control children. Children with PKU performed more poorly than typically developing children on the 2 inhibitory tasks with the strongest experimental manipulations (go/no-go and antisaccade) between control and inhibitory conditions. Findings suggest that the inhibitory deficit associated with PKU is subtle and that inconsistent findings in past studies may be largely due to the insensitivity of experimental manipulations in some tasks. PMID- 17083297 TI - Cluster subtypes on the California verbal learning test-children's version after pediatric traumatic brain injury. AB - The purpose of this study is to determine the presence of profile subtypes on the California Verbal Learning Test-Children's Version (CVLT-C; Delis, Kramer, Kaplan, and Ober, 1994) in 175 children with traumatic brain injury (TBI). Four key z score variables are used in a 2-stage cluster analysis that reveal 4 reliable subtypes. No meaningful differences among the clusters are found on demographic variables. In contrast, statistically significant differences among the 4 clusters in both level and pattern of performance are found on injury severity parameters and the 4 factor index scores from an independent measure of psychometric intelligence (Wechsler Intelligence Scale for Children-Third Edition; Wechsler, 1991). This study concludes that although no unique profile is found on the CVLT-C after TBI in children, performance on this test is affected strongly by injury severity, with a mediating contribution by speed of information processing. PMID- 17083298 TI - Naming speed and visual search deficits in readers with disabilities: evidence from an orthographically regular language (Italian). AB - The study examined rapid automatized naming (RAN) in 42 children with reading disabilities and 101 control children-all native speakers of Italian, a language with shallow orthography. Third-, 5th- and 6th-grade children were given a RAN test that required rapid naming of color, object, or digit matrices. A visual search test using the same stimulus material (but not requiring a verbal response) and an oral articulation test were also given. Readers with disabilities performed worse than controls on the RAN test. This effect was larger in higher grades than in lower ones. Readers with disabilities were also slower than controls in performing the visual search test. The pattern of results for the RAN test held constant when the visual search performance was partialed out by covariance analysis, indicating the independence of the 2 deficits. The 2 groups did not differ for articulation rate. Finally, analysis of the pattern of intercorrelations indicated that reading speed was most clearly related to RAN, particularly in the group with reading disabilities. The results extend observations of RAN effects on reading deficits to Italian, an orthographically shallow language. PMID- 17083299 TI - Learning disabilities and intellectual functioning in school-aged children with prenatal cocaine exposure. AB - Risk for developing a learning disability (LD) or impaired intellectual functioning by age 7 was assessed in full-term children with prenatal cocaine exposure drawn from a cohort of 476 children born full term and enrolled prospectively at birth. Intellectual functioning was assessed using the Wechsler Intelligence Scale for Children-Third Edition (Wechsler, 1991) short form, and academic functioning was assessed using the Wechsler Individual Achievement Test (WIAT; Wechsler, 1993) Screener by examiners blind to exposure status. LDs were categorized based on ability-achievement discrepancy scores, using the regression based predicted achievement method described in the WIAT manual. The sample in this report included 409 children (212 cocaine-exposed, 197 non-cocaine-exposed) from the birth cohort with available data. Cumulative incidence proportions and relative risk values were estimated using STATA software (Statacorp, 2003). No differences were found in the estimate of relative risk for impaired intellectual functioning (IQ below 70) between children with and without prenatal cocaine exposure (estimated relative risk = .95; 95% confidence interval [CI] = 0.65, 1.39; p = .79). The cocaine-exposed children had 2.8 times greater risk of developing a LD by age 7 than non-cocaine-exposed children (95% CI = 1.05, 7.67; p = .038; IQ >/= 70 cutoff). Results remained stable with adjustment for multiple child and caregiver covariates, suggesting that children with prenatal cocaine exposure are at increased risk for developing a learning disability by age 7 when compared to their non-cocaine-exposed peers. PMID- 17083301 TI - Do patients with sleep maintenance insomnia have a problem with sleep maintenance? AB - Do patients with primary insomnia differ from good sleepers with respect to the number or duration of awakenings or to the stages from which awakenings occur? To address this question, polysomnography (PSG) records were evaluated in 10 good sleepers (GS) and 10 primary insomnia patients (PI). PSG records were evaluated for occurrence and duration of awakenings and for the stage immediately preceding each awakening. PIs woke more frequently and for longer durations than did GSs. PIs' awakenings tended to occur from Stages 1 or 2; GSs' occurred from epochs scored as movement times. The data from this study represent the first attempt to characterize the stages from which awakenings occur in sleep maintenance insomnia. PMID- 17083302 TI - The effect of sleep-disordered breathing on stages of sleep in patients with Alzheimer's disease. AB - Patients with Alzheimer's disease (AD) commonly have poor sleep and a high reported incidence of sleep-disordered breathing (SDB). This study examined how the presence of SDB affected sleep stages in AD patients. Sixty-six volunteers with mild - moderate AD underwent home polysomnography. Results showed that patients with SDB spent less of the night in REM sleep than those with no SDB, but there were no differences in other sleep stages. The findings suggest that the decreased amount of REM sleep may be due to the presence of AD and SDB. Treating these patients' SDB may increase their amount of REM sleep, which may result in improved daytime functioning. Controlled trials of SDB treatment in AD are needed to answer this question. PMID- 17083303 TI - Symptom-focused rumination and sleep disturbance. AB - Rumination can prolong negative mood, disrupt sleep, and increase depression risk. Although there is evidence that poor sleepers ruminate, no studies have identified the ruminative content relevant for sleep disturbance. This study investigated (a) the association between rumination and sleep and (b) the ruminative content of poor sleepers. Results revealed that self-defined poor sleepers (n = 104) were more prone than self-defined good sleepers (n = 139) to ruminate and that the ruminative content was symptom focused (e.g., poor sleepers ruminated on causes of dysphoria, concentration, and fatigue symptoms). As dysphoria, reduced concentration, and fatigue are all commonly experienced daytime symptoms of insomnia, this preliminary finding of symptom-focused rumination should be further evaluated as a risk factor for further sleep disturbance in clinical samples as well as a possible link between insomnia and depression. PMID- 17083304 TI - Effects of an intervention aimed at reducing night waking and signaling in 6- to 12-month-old infants. AB - This quasi-experimental one-group pre- and posttest pilot study evaluated an intervention aimed at reducing night waking and signaling for infants between 6 and 12 months of age. Thirty-nine healthy infants and their parents were recruited. Thirty-five infants completed the intervention and data collection. Both parents participated in a group teaching session with telephone follow-up for 2 weeks. Actigraphy and sleep diary data were collected at baseline and 6 and 16 weeks postintervention. We hypothesized a decrease in length and number of infant waking and crying periods and an increase in longest night sleep and nap time. Following the intervention, infants had significantly reduced length of night crying and number of wakes and longer night sleep periods. The intervention warrants evaluation with a randomized controlled design. PMID- 17083306 TI - Sweeping changes, or a sweep under the rug? PMID- 17083307 TI - Influence of broad-spectrum antibiotic prophylaxis on intracranial pressure monitor infections and subsequent infectious complications in head-injured patients. AB - BACKGROUND: The benefit of antibiotic prophylaxis for intracranial pressure (ICP) monitors remains controversial, and clinical practice varies widely. Whether any antibiotic coverage, particularly broad-spectrum coverage, reduces monitor related infections remains unproved, and exposure to antibiotics may affect the susceptibility patterns of pathogens producing subsequent infectious complications. Despite the lack of data supporting its use, our level I trauma center had a long-standing ICP monitor prophylaxis protocol that provided broad spectrum coverage that included ceftriaxone. In April 2002, a protocol change was instituted that substituted cefazolin for ceftriaxone as single-agent prophylaxis for ICP monitors. HYPOTHESIS: Broader-spectrum antibiotic prophylaxis does not reduce ICP monitor-related infections but is associated with acquisition of more drug-resistant infections than narrow-spectrum prophylaxis. METHODS: To evaluate the influence of broad- versus narrow-spectrum prophylaxis, a three year period encompassing each practice was selected. All injured patients with ICP monitors placed between January 1, 2001, and December 31, 2003 (n = 279), were identified using the Vanderbilt trauma database. Antibiotic prophylaxis for ICP monitors was determined using the hospital financial database to identify all antibiotics given to individual patients and subsequent chart review to identify those antibiotics given solely for ICP prophylaxis. A total of 119 patients received narrow-spectrum (either cefazolin or vancomycin; n = 100) or no (n = 19) prophylaxis, whereas 160 received broad-spectrum prophylaxis (ceftriaxone or ciprofloxacin). The two groups did not differ with respect to baseline demographics, type of ICP monitor, or duration of monitor placement. Infectious complications were determined by continuous infection surveillance utilizing standard U.S. Centers for Disease Control and Prevention National Nosocomial Infection Surveillance System (CDC-NNIS) definitions and maintained in a contemporary database. The influence of broad-spectrum antibiotic prophylaxis on both ICP monitor infections and subsequent infections outside the central nervous system (CNS) was determined. RESULTS: Nine patients (3.2%) developed CNS infections; two of 119 patients (1.7%) who received narrow-spectrum or no prophylaxis versus seven of 160 patients (4.4%) who received broad-spectrum prophylaxis (p = NS). Only the duration of monitor placement and Injury Severity Score were associated with the infection rate. In the total population, 185 infections occurred in 93 patients (33%). Infection rates did not differ between patients who received narrow-spectrum or no prophylaxis (32%) and those who received broad-spectrum prophylaxis (34%). However, patients who received broad spectrum prophylaxis acquired gram-negative infections with significantly greater antibiotic resistance. CONCLUSIONS: Broad-spectrum antibiotic prophylaxis of ICP monitors does not reduce CNS infections, but is associated with a shift to resistant gram-negative pathogens in subsequent infectious complications. Thus, broad-spectrum antibiotic prophylaxis of ICP monitors should be eliminated or minimized unless data from randomized trials prove its utility. PMID- 17083308 TI - Can we define the ideal duration of antibiotic therapy? AB - BACKGROUND: Because of the increasing development of antimicrobial resistance, there is a greater responsibility within the medical community to limit the exposure of patients to antibiotics. We tested the hypothesis that shorter courses of antibiotics are associated with similar or better results than longer durations. We also sought to investigate the difference between a fixed duration of therapy and one based on physiologic measures such as fever and leukocytosis. METHODS: All infectious episodes on the general surgery units of the University of Virginia Health System from December 15, 1996, to July 31, 2003, were analyzed retrospectively for the relation between the duration of antibiotic therapy and infectious complications (recurrent infection with the same organism or at the same site). All infections associated with either fever or leukocytosis were categorized into quartiles on the basis of the absolute length of antibiotic administration or the duration of treatment following resolution of fever or leukocytosis. Multivariate logistic regression models were developed to estimate the independent risk of recurrence associated with a longer duration of antibiotic use. RESULTS: Of the 5,561 treated infections, 4,470 were associated with fever (temperature > or =38 degrees C) or leukocytosis (white blood cell count > or =11,000/mm(3)). For all infections, whether analyzed by absolute duration or time from resolution of leukocytosis or fever, the first or second quartiles (0-12 days, 0-9 days, 0-9 days, respectively) were associated with the lowest recurrence rates (14-18%, 17-23%, 18-19%, respectively). Individual analysis of intra-abdominal infections and pneumonia yielded similar results. The fixed-duration groups received fewer days of antibiotics on average, with outcomes similar to those in the physiologic parameters group. CONCLUSIONS: Shorter courses of antibiotics were associated with similar or fewer complications than prolonged therapy. In general, adopting a strategy of a fixed duration of therapy, rather than basing duration on resolution of fever or leukocytosis, appeared to yield similar outcomes with less antibiotic use. PMID- 17083309 TI - The open abdomen in trauma: do infectious complications affect primary abdominal closure? AB - OBJECTIVE: One of the primary goals of damage control surgery in the trauma patient is primary closure of the abdomen. We hypothesized that extra-abdominal infections, such as those complicating injuries to the thorax, diaphragm, long bones, or musculoskeletal system, would decrease the likelihood of primary abdominal closure and increase hospital resource utilization in patients requiring open abdominal management. METHODS: The trauma registry of the American College of Surgeons (TRACS) was reviewed retrospectively from 1995-2002 for open abdomen technique and damage control surgery. The outcome was primary fascial closure or delayed closure. Patients who died prior to closure were excluded. We evaluated infectious complications, including ventilator-associated pneumonia (VAP), blood stream infection (BSI), and surgical site infection (SSI). Other parameters studied were multiple rib fractures, long bone fractures, chest injuries, diaphragm injuries, empyema, and transfusion requirements. Hospital charges were obtained from the hospital administrative database. Univariate, multivariate, and regression analyses were performed to identify the effects of infectious complications on primary abdominal closure, length of stay, total hospital charges, and disposition. RESULTS: Three hundred forty-four patients required the open abdomen technique: 67% received damage control laparotomy and 33% decompression of abdominal compartment syndrome. Two hundred seventy-six patients (80%) went on to abdominal closure of some form and constituted the primary study group. Primary abdominal closure was achieved in 180 (65%) with a mean time to closure of 3.5 days. Ventilator-associated pneumonia, BSI, and SSI were associated with lack of primary closure (p < 0.05). Increased blood transfusions also were associated with failure of primary closure (p < 0.05). Ventilator-associated pneumonia and BSI were associated with significantly greater lengths of stay in the intensive care unit (ICU) (24.2 days vs. 12.6 days and 30.5 days vs. 17.9 days; both p < 0.0001) and significantly greater total hospital charges (232,080 US dollar vs. 142,893 US dollar; 247,440 US dollar vs. 160,940 US dollar; and 264,778 US dollar vs. 170,447 US dollar; all p < 0.001). CONCLUSION: Inability to achieve primary abdominal closure was associated with infectious complications (VAP, BSI, and SSI) and large transfusion requirements. Infectious complications also significantly increased ICU utilization and hospital charges. Death was associated with BSI, femur fractures, and large transfusion requirements, whereas infectious complications did not have a significant impact on discharge disposition. PMID- 17083310 TI - Variable mannose-binding lectin expression during postoperative acute-phase response. AB - BACKGROUND: Low plasma concentrations and genetic polymorphisms of mannan-binding lectin (MBL) have been associated with infectious disease complications during various conditions. The present study examined the nature and expression of MBL deficiency during a surgery-induced acute-phase response. METHODS: Blood was sampled from 20 consecutive patients before and 1, 3, 5, 7, and 10 days and 6 weeks after a uniform abdominal operation (transhiatal esophagectomy). Plasma concentrations of MBL, C-reactive protein (CRP), and secretory phospholipase A2 (sPLA2) were measured. Patients were classified as low- or high-level MBL producers by their preoperative concentration (<0.5 or > or = 0.5 micrograms/mL), and were cross-verified for actual MBL deficiency by nucleotide sequencing of both the MBL promoter and exon-1 alleles. RESULTS: Baseline plasma MBL concentrations correlated with maximal postoperative plasma concentrations (r = 0.88; p < 0.0001). This was not found for CRP and sPLA2 (r = 0.19 and r = 0.08, respectively). Alleles responsible for structural MBL variants were detected in 40% of patients and were associated with significantly reduced MBL concentrations (p = 0.005). The baseline cut-off value in plasma of 0.5 micrograms/mL clearly identified individuals with variant exon-1 alleles (sensitivity 100%, specificity 83%). CONCLUSIONS: Baseline MBL plasma concentrations are predictive of MBL expression during the acute-phase response. A baseline cut-off value of 0.5 micrograms/mL can be used to identify patients with variants in the exon-1 region of the MBL gene without the need for nucleotide sequencing. Clinical studies may use this easy and quick method to identify MBL deficient patients preoperatively, as they are conditionally at risk for infectious complications. PMID- 17083311 TI - Decision-making analysis for selection of antibiotic treatment in intra-abdominal infection using preference measurements. AB - BACKGROUND: Antimicrobial therapy of abdominal infections is important to the prognosis of affected patients. The choice of antimicrobial therapy must consider effectiveness, safety, cost, and antibiotic resistance, among numerous factors. However, in reality, decisions are made assuming bioequivalence between regimens, without considering the specific attributes of any particular regimen. The objective was to determine the best antibiotic regimen for patients with community-acquired abdominal infection on the basis of a decision analysis that included effectiveness as well as safety, measured as adverse effects. METHODS: A decision tree was built using information from a systematic review of the literature on the effectiveness of antimicrobial regimens tested in randomized clinical trials (RCTs) and the frequency and severity of adverse effects. The quality of the articles was assessed with the Oxford criteria for RCTs. The main outcome was preferences reported by surgeons, measured on a numeric scale. Preferences were obtained using a standard survey that reported each adverse effect with its respective intensity, reversibility, sequelae, duration of symptoms, and necessity for change of antibiotic. Each of the surgeons had to assign a value blindly from 0 to 10, where 10 was the most severe. A sensitivity analysis was conducted varying the frequency of adverse effects. RESULTS: The regimens analyzed were amikacin-metronidazole, amikacin-clindamycin, ciprofloxacin-metronidazole, ampicillin-sulbactam, ceftriaxone-metronidazole, piperacillin-tazobactam, and ertapenem. The perceived severity of adverse effects reported were: Acute neuromuscular blockade (8.0), severe allergic reaction (7.5), ototoxicity (7.4), nephrotoxicity (7.1), antibiotic-associated colitis (7.0), peripheral neuropathy (5.3), general neurological symptoms (4.9), gastrointestinal symptoms (3.1), and other general symptoms (2.6). Favored regimens were ceftriaxone-metronidazole (1.15), ampicillin-sulbactam (1.24), piperacillin-tazobactam (1.27) and ertapenem (1.28). These strategies dominated the other therapeutic schemes. Sensitivity analysis showed no changes in the dominance reported when the frequency of adverse effects was maintained in the known clinical range. CONCLUSIONS: Antibiotic regimens that contain aminoglycosides are not bioequivalent to those without aminoglycosides when effectiveness and adverse effects are considered simultaneously. Antibiotic regimens that do not use aminoglycosides must be the first line of treatment for abdominal sepsis acquired in the community. PMID- 17083312 TI - Preoperative concentrations of suPAR and MBL proteins are associated with the development of pneumonia after elective surgery for colorectal cancer. AB - BACKGROUND: The urokinase-type plasminogen activation (uPA) system and the mannan binding lectin (MBL) complement activation pathway are involved in regulation of immune responses. The blood concentrations of these molecules in the individual patient thus could be related to the risk of postoperative infectious complications. The aim of this retrospective study was to determine the association between the soluble uPA receptor (suPAR) and MBL concentrations and the development of postoperative bacterial infectious complications. METHODS: Blood samples were drawn preoperatively from 544 patients scheduled to undergo primary resection for colorectal cancer. Plasma suPAR was determined by enzyme linked immunosorbent assay and serum MBL by time-resolved immunofluorescent assay. The following infectious events were recorded during the first month after surgery: surgical site or perineal infection or both, intra-abdominal abscess, anastomotic leakage, pneumonia, and blood stream infection. Data on perioperative blood transfusions in addition to clinical baseline characteristics were included as well. RESULTS: The numbers of surgical site infections, intra-abdominal abscesses, anastomotic leakages, pneumonias, and blood stream infections were 51, 20, 32, 78, and 19, respectively. Univariate analysis showed that elevated concentrations of suPAR (p = 0.01; odds ratio [OR] 2.2; 95% confidence interval [CI] 1.2, 3.9), low concentrations of MBL (p = 0.047; OR 0.9; 95% CI 0.8, 1.0), and perioperative blood transfusion (p = 0.006; OR 1.5; 95% CI 1.1, 2.0) were associated with the development of pneumonia. Significant associations with other bacterial infections could not be demonstrated. Multivariate analysis including disease stage, sex, and age showed that suPAR, MBL, and perioperative blood transfusion were significantly and independently associated with postoperative pneumonia. CONCLUSION: Concentrations of suPAR and MBL, in addition to perioperative blood transfusion, were significantly associated with the development of postoperative pneumonia. No other statistically significant relationships could be demonstrated. Thus, further research should be directed to clarifying the biological role of these two molecules in the development of postoperative pneumonia. PMID- 17083313 TI - The obese surgical patient: a susceptible host for infection. AB - BACKGROUND: Obesity is common in the Western world, and obese persons constitute a growing population of surgical patients for both bariatric and non-bariatric operations. It is the traditional perception that obese patients have a higher risk of perioperative morbidity and mortality, although different studies show contradictory results. PURPOSE: To better delineate the perioperative morbidity and mortality in obese patients. METHODS: Review of the pertinent English language literature RESULTS: Obesity is a risk factor for nosocomial infection, particularly surgical site infection (SSI). The mechanisms by which obese patients may be at higher risk for SSI are reviewed, and specific recommendations are outlined that should be implemented when treating obese patients to minimize potentially preventable SSIs. CONCLUSION: The growing prevalence of obesity and the increasing number of operations performed on obese patients, whether to achieve weight loss or for other purposes, will have a substantial impact on health care resources. Vigilant identification of high-risk patients and provision of all proved preventive measures must suffice until new methods of prevention are identified and validated. PMID- 17083314 TI - Do marking inks pose an infection risk? A surgeon's perspective. PMID- 17083315 TI - Mixed methods research for the novice researcher. AB - Mixed methods research is becoming increasingly popular in the health and social science disciplines. The aim of this article is to give an overview of the varieties of mixed methods designs. We begin by situating mixed methods research in the context of a paradigmatic framework which assists a researcher in making decisions concerning the design of their study. Although the most commonly used mixed methods designs are underpinned by positivist/postpositivist assumptions, the combination of qualitative and quantitative methods can be used within any research paradigm. PMID- 17083316 TI - The nurse academic in the clinical learning setting: an evolving partnership role. AB - Student readiness for practice as a registered nurse can only be achieved through a committed and cooperative partnership between the tertiary and clinical sectors. One important role in the development and maintenance of clinical learning partnerships is that of the nurse academic. As nursing faculties continue to evolve this role it is important that positive initiatives be shared and challenges discussed. With this in mind, the role of the nurse academic in clinical practice introduced at the University of Technology, Sydney (UTS) will be presented. A nurse academic who fulfils this role at UTS is known as an Academic Liaison Person (ALP). This paper begins with a general exploration of the role of the academic in the clinical learning setting and then discusses the associated challenges. The role of the ALP at UTS and its integration with the UTS clinical model for final year students will be presented as a response to these challenges. Through this model the educational expertise of the ALP and the clinical expertise of the practitioner is brought together for the benefit of the nursing student. PMID- 17083317 TI - Communication and decision making for patients with end stage diseases in an acute care setting. AB - Twenty retrospective patient case studies were collated in an acute care teaching hospital using a case note audit and in addition interviews were undertaken with 40 nursing staff following the deaths of these patients in order to: analyse the end of life care received; identify any deficits in care provision and to enable the nursing division to target any inadequacies in care found. Findings indicated that communication between medical and nursing staff and between nursing staff, patients and family around end of life issues continue to be poor and that discussions regarding NFR decisions occurred too close to death, creating unnecessary stress for both patients and families. Recommendations regarding palliative approaches in the acute care setting are detailed. PMID- 17083318 TI - Turkish version of the Jefferson Scale of Attitudes Toward Physician-Nurse Collaboration: a preliminary study. AB - BACKGROUND: The Jefferson Scale of Attitudes Toward Physician-Nurse Collaboration is an established instrument to measure the perceived work relationship between physicians and nurses. The survey addresses areas of autonomy and decision making, interprofessional education and relations, psychosocial care, teamwork, and shared responsibility. The aim of this prelimiary study was to adapt the Jefferson Scale of Attitudes Toward Physician-Nurse Collaboration for use in Turkey and test its psychometric properties and utility in clinical and research practice. METHODS: The process of cross-cultural adaptation and validation followed the guidelines provided in the existing literature. First, three bilingual health professionals independently translated the original questionnaire into Turkish and a consensus version was generated. Then, three other translators, blind to the original questionnaire, performed a back translation into English to confirm the accuracy of the translation. This version was then compared with the original English questionnaire. Discrepancies were discussed and solved by a panel of two nurses and two physicians. The field testing for face validity was done in a group of ten monolingual physicians and nurses. Reliability was assessed with test-retest reliability and construct validity was confirmed with factor analysis. RESULTS: The mean time of questionnaire administration was 3 minutes and 45 seconds. The test-retest reliability was 0.75, and Cronbach's coefficient alpha was 0.71 for the entire sample. The findings of the factor analysis indicated that the Turkish version of Jefferson Scale of Attitudes Toward Physician-Nurse Collaboration was comprised of the following four factors: 'physician-nurse relationships', 'shared education', 'nursing role in patient care', and 'accountability and responsibility of nurses.' CONCLUSION: The overall findings of this study indicate that the Turkish version of the Jefferson Scale of Attitudes Toward Physician-Nurse Collaboration is a psychometrically sound tool with satisfactory measurement characteristics including construct validity and internal consistency reliability. This instrument may be useful in assessing the effectiveness of educational programs designed to enhance collaboration between physicians and nurses, whether these programs are aimed at residents and graduate nursing students or practicing physicians and nurses. PMID- 17083319 TI - The role of the breast care nurse during treatment for early breast cancer: the patient's perspective. AB - INTRODUCTION: Women who are diagnosed with breast cancer require the support of a range of people during their treatment. Although the role of the breast care nurse in providing support has recently been investigated in several Australian studies the patients' perspective on the role of the breast care nurse in Australian hospitals has not previously been described in detail. The aim of this paper is to explore patients' perspectives on the role of the breast care nurse. METHODS: In-depth interviews were conducted with 18 women who had completed treatment for early breast cancer. The women were asked to describe their experiences from the time of diagnosis through to treatment completion. Thematic analysis was used to analyse the data. FINDINGS: Breast cancer patients repeatedly emphasised the importance of the role of their breast care nurses throughout their experience of breast cancer. The support that breast care nurses provided incorporated the following components: communication, rapport and an awareness of the women's needs, availability, reassurance and practical information. CONCLUSION: This study provides an understanding of women's perspectives on the role of breast care nurses and confirms that breast care nurses play an important support role during the experience of early breast cancer. Oncology clinics should focus on ensuring that all women who are diagnosed with breast cancer have adequate access to the support that breast care nurses are able to provide. PMID- 17083320 TI - Enhancing nursing students' clinical placement experiences: a quality improvement project. AB - Clinical experience is recognised as the core of nursing education. Quality clinical placements across a variety of venues are vital to the development of capable and competent professionals. However there is evidence, both anecdotal and empirical, suggesting that students' clinical placement experiences are fraught with problems. The quality improvement project described in this paper aimed to improve the clinical learning experience of nursing students by strengthening communication and partnerships between the university and the two local health services. To achieve this goal, clinicians' perceptions of the problems related to clinical placements and their recommendations for improvement were explored. Focus groups, brainstorming sessions, personal interviews and surveys were used to collect qualitative and quantitative data. Problems and priority issues were identified as more than five hundred clinicians expressed their concerns, claims and issues. Key findings from the project are described under five themes: communication breakdown between the university and clinicians; mentorship; preparation for clinical placements; clinical competence; and graduates' readiness for practice. Utilising a quality improvement framework this project promoted vigorous debate and dialogue between university and health service partners. The nature and extent of the problems surrounding clinical placements were examined, high priority issues targeted for improvement, and the subsequent results measured through feedback from clinicians and students. The success of this project, although impressive in the early stages, will depend upon ongoing communication and evaluation to ensure sustainability of the improvements made. PMID- 17083321 TI - 'You find yourself.' Perceptions of nursing students from non-English speaking backgrounds of the effect of an intensive language support program on their oral clinical communication skills. AB - Nurses of ethnically diverse backgrounds are essential in providing multicultural populations in western societies with culturally and linguistically competent health care. However, many nurses from non-English speaking backgrounds (NESB) are at high risk of failure in university programs particularly during clinical placements. Few studies investigate the clinical experiences of students from NESB and strategies to support their learning. This study describes perceptions of fifteen undergraduate nursing students from NESB about their first clinical placement in an Australian university program and the effect of a language support program on their oral clinical communication skills. Three categories arose: *Wanting to belong but feeling excluded; *Wanting to learn how to...; and *You find yourself. While many students find clinical placement challenging, it appeared difficult for students in this study as language and cultural adjustments required some modification of their usual ways of thinking and communicating, often without coping strategies available to other students. PMID- 17083322 TI - Which way? Educating for nursing Aboriginal and Torres Strait Islander peoples. AB - Cross-Cultural Awareness Training has been seen as a way to improve nurses' knowledge and understanding of Indigenous peoples in Australia (Aboriginal and Torres Strait Islanders) and to therefore improve service delivery and therapeutic care to them. Nurses may have undertaken this type of training in their workplace or as part of nurse education in an undergraduate degree program. In asking Which Way in regards to this type of training and education, this paper includes the views of a selection of Aboriginal women and highlights the need to extend beyond Cross-Cultural Awareness Training to Anti-Racism Training. Furthermore, that Anti-Racism Training and addressing white race privilege is required in order to address the inequities within the health system, the marginalisation and disempowerment of Indigenous peoples. PMID- 17083323 TI - An exploration of modifiable health associated risk factors within a cohort of undergraduate nursing students. AB - So-called diseases of affluence, otherwise known as 'lifestyle diseases', are attributed to modifiable risk factors that are influenced by lifestyle and personal behaviour. Leading by example is an important way for public health principles to be communicated. In the university context, students of nursing can become aware of the challenge to integrate and apply health principles in their own life so that they become responsible health leaders in the community. The aim of this study was to explore the incidence of a number of behaviour-associated health risk factors within a group of undergraduate nursing students. Ninety-four students participated in the study. Seventy-seven students (82%) reported the presence of at least one modifiable health risk factor. Forty-four percent of respondents were either overweight or obese. Further research to explore whether a health promoting curriculum encourages nursing students to internalise/apply health knowledge to their own lives is recommended. A campaign of public health might be useful within the university community to educate students about risk factors and healthy living. PMID- 17083325 TI - Positioning mentorship within Australian nursing contexts: a literature review. AB - There are a variety of structured and unstructured supportive relationships available to nurses. Internationally, nurses commonly use preceptorship, clinical supervision, and mentorship to meet distinct needs and provide differing levels of commitment, intensity, and enabling functions. Of particular interest to the nursing profession is the use of mentoring relationships to support nurses in achieving leadership positions. In Australia, preceptorship and clinical supervision are freely used and understood by nurses however, mentoring relationships are less readily applied, and agreed meanings and understanding are lacking. This paper will explore the range of supportive relationships available to nurses. The terms used to define and describe these relationships, and how these relationships are contextualised, will be explored in order to better understand the position of mentorship for nurses in Australia. The potential value of mentorship in developing nursing leadership in Australia will also be identified. PMID- 17083324 TI - Improving the implementation of an early literacy program by child health nurses through addressing local training and cultural needs. AB - This study evaluates the implementation of an early literacy program by child health nurses (CHNs) in Geraldton and the Midwest regions of Western Australia. Children's early preschool exposure to reading is an important step in developing literacy skills in later schooling. The program aims to improve early literacy development by supplying parents with an appropriate book and information on reading to children at the child's seven-to-nine months of age health check. This study highlights the potential role of CHNs in encouraging early literacy in regional communities. However, a barrier to the successful implementation of the program was found to be its dependence on attendance at a predetermined health check. This was especially so for the Aboriginal population, which had a significantly lower attendance rate. Dependence on personal and professional experience amongst CHNs also resulted in sub-optimal implementation of the program. Recommendations to broaden the scope of the intervention, in timing and location, and to develop appropriate training for CHNs are discussed. It is shown that the implementation of such evidence-based programs must address local community and service provider issues to reach their full potential. PMID- 17083326 TI - Person-centredness: a concept analysis. AB - Person-centred care is a term that has been used frequently in literature since the mid-1990s. This type of care requires health care staff to use person centredness as a focus for developing relationships and plans of care. In an attempt to define person-centredness, a literature review was undertaken as a means of developing a concept analysis. This led to profiling attributes of person-centredness developed from the work of early theorists and authors as they began to recognise the importance of the ethical and legal rights of people and the importance of holistic care in maintaining wellbeing. However, in the literature, the term person-centred was often interchanged with terms such as patient or client-centred which may lead to confusion. This concept analysis shows that many of the attributes of person-centredness are indeed relevant to patient and client centred care. However, the connotations behind client and patient may appear to shift the balance of power to the health carer by concentrating on the illness rather than the person. Therefore, clarification of the term person-centredness may assist future researchers, practitioners and authors to apply one term only until evolution leads to further definition or change. PMID- 17083328 TI - Identification of a novel nucleoside triphosphatase from Mycoplasma mobile: a prime candidate motor for gliding motility. AB - A protein with a molecular mass of 42 kDa (P42) from Mycoplasma mobile, one of several mycoplasmas that exhibit gliding motility, was shown to be a novel NTPase (nucleoside triphosphatase). Although the P42 protein lacks a common ATP-binding sequence motif (Walker A), the recombinant proteins expressed in Escherichia coli certainly hydrolysed some nucleoside triphosphates, including ATP. The results of photoaffinity labelling by an ATP analogue supported that the P42 protein contains a specific binding site for ATP (or another nucleoside triphosphate). In the M. mobile genome, the P42 gene is located downstream of gli123, gli349 and gli521 genes, and they have been reported to be polycis-tronically transcribed. As the huge proteins encoded by gli123, gli349 and gli521 play a role in gliding motility of M. mobile, P42 might also have some kind of function in the gliding motility. The gliding motility of M. mobile is driven directly by ATP hydrolysis, but the key ATPase has not been identified. Our results showed that, among these four proteins, only P42 exhibited ATPase activity. Biochemical characteristics- optimal conditions for activity, substrate specificities, and inhibiting effects by ATP analogues--of the recombinant P42 proteins were very similar to those of a putative ATPase speculated from a previous analysis with a gliding 'ghost' whose cell membrane was permeabilized by Triton X-100. These results support the hypothesis that the P42 protein is the key ATPase in the gliding motility of M. mobile. PMID- 17083329 TI - Histone acetyltransferase MOZ acts as a co-activator of Nrf2-MafK and induces tumour marker gene expression during hepatocarcinogenesis. AB - HATs (histone acetyltransferases) contribute to the regulation of gene expression, and loss or dysregulation of these activities may link to tumorigenesis. Here, we demonstrate that expression levels of HATs, p300 and CBP [CREB (cAMP-response-element-binding protein)-binding protein] were decreased during chemical hepatocarcinogenesis, whereas expression of MOZ (monocytic leukaemia zinc-finger protein; MYST3)--a member of the MYST [MOZ, Ybf2/Sas3, Sas2 and TIP60 (Tat-interacting protein, 60 kDa)] acetyltransferase family--was induced. Although the MOZ gene frequently is rearranged in leukaemia, we were unable to detect MOZ rearrangement in livers with hyperplastic nodules. We examined the effect of MOZ on hepatocarcinogenic-specific gene expression. GSTP (glutathione S-transferase placental form) is a Phase II detoxification enzyme and a well-known tumour marker that is specifically elevated during hepatocarcinogenesis. GSTP gene activation is regulated mainly by the GPE1 (GSTP enhancer 1) enhancer element, which is recognized by the Nrf2 (nuclear factor erythroid 2 p45 subunit-related factor 2)-MafK heterodimer. We found that MOZ enhances GSTP promoter activity through GPE1 and acts as a co-activator of the Nrf2-MafK heterodimer. Further, exogenous MOZ induced GSTP expression in rat hepatoma H4IIE cells. These results suggest that during early hepatocarcinogenesis, aberrantly expressed MOZ may induce GSTP expression through the Nrf2-mediated pathway. PMID- 17083330 TI - FVB.129P2-Pde6b(+) Tyr(c-ch)/Ant, a sighted variant of the FVB/N mouse strain suitable for behavioral analysis. AB - Mice of the FVB/N strain are severely visual impaired as a result of tyrosinase gene defects, leading to a deficiency of the key enzyme for melanin synthesis in skin and eye and of cyclic guanosine monophosphate phosphodiesterase gene defects, which results in albinism (Tyr(c/c)) and retinal degeneration (Pde6b(rd1/rd1)), respectively. Nevertheless, FVB/N mice are commonly used for the generation of transgenic animals because of their large, strong pronuclei and high breeding performance. However, due to visual impairment of the FVB/N animals, the resulting transgenic animals cannot be used in tests that depend on vision, including tests of cognitive behavior. Therefore, we have bred a sighted version of the FVB/N strain by an outcross between FVB/N and 129P2/OlaHsd, followed by repeated backcrosses to FVB/N mice while selecting against albinism and homozygosity of the retinal degeneration mutation. After 11 generations of backcrossing, sighted animals were intercrossed to generate the congenic FVB.129P2-Pde6b(+) Tyr(c-ch)/Ant strain, which is pigmented (Tyr(c-ch)/(c-ch)) and devoid of the genetic predisposition to retinal degeneration. The accurate visual abilities of the FVB.129P2-Pde6b(+) Tyr(c-ch)/Ant mice, for which we propose the name FVBS/Ant, demonstrated a clear visual evoked potential in the presence of normal eye histology and improved performance in the Morris water maze test. PMID- 17083331 TI - Vasopressin 1a receptor knockout mice have a subtle olfactory deficit but normal aggression. AB - Two receptors for vasopressin (Avp) are expressed in the brain, the Avp 1a receptor (Avpr1a) and the Avp 1b receptor (Avpr1b). To investigate the role of Avpr1a in behaviors in mice more extensively, we generated a line of mice lacking a functional Avpr1a (knockout, Avpr1a(-/-)). We first performed a baseline phenotypic screen of the Avpr1a knockouts followed by a more detailed analysis of their circadian rhythms and olfactory function. When free-running in constant darkness, the Avpr1a(-/-) mice have a longer circadian tau than the wild types. There are also subtle olfactory deficits in Avpr1a(-/-) mice as measured in an olfactory habituation/dishabituation test and in the discrimination of female urine from male urine using an operant testing paradigm. An extensive body of research has shown that manipulation of the Avpr1a alters behavior, including aggression and social recognition. Therefore, we expected profound behavioral deficits in mice lacking the Avpr1a gene. Contrary to our expectations, social aggression, anxiety-like behavior and social recognition are unaffected in this line of Avpr1a knockout mice. These data suggest either that the Avpr1a is not as critical as we thought for social behavior in mice or, more likely, that the neural circuitry underlying aggression and other social behaviors compensates for the life-long loss of the Avpr1a. However, the olfactory deficits observed in the Avpr1a(-/-) mice suggest that Avp and Avpr1a drugs may affect behavior, in part, by modulation of chemosensory systems. PMID- 17083332 TI - Social isolation and expression of serotonergic neurotransmission-related genes in several brain areas of male mice. AB - Early-life events influence brain development and evoke long-lasting behavioral consequences. Postweaning social isolation in rodents induces emotional and neurochemical alterations similar to those observed among some human psychopathologies. Central serotonergic neurotransmission is intimately involved in the observed adjustments, but the impact of social deprivation on serotonergic gene expression is unknown. We investigated the effects of prolonged early social isolation on emotion-related behaviors and 5-hydroxytryptamine (5-HT)-related gene transcription in mice. After weaning, male C57BL/6J mice were reared singly or in groups of four for 6 weeks. Gene expression of 5-HT(1A), 5-HT(1B), 5 HT(2A), 5-HT(2C), 5-HT(3A), 5-HT(6) and 5-HT(7) receptors and of 5-HT transporter and tryptophan hydroxylase-2 was determined by quantitative real-time polymerase chain reaction in distinct brain areas. Single-housed mice were hyperactive in a novel environment and showed signs of aggressive behavior. Housing condition did not alter weight gain or body temperature. Isolation markedly reduced transcription of all postsynaptic 5-HT receptors in the prefrontal cortex and reduced 5-HT(1B), 5-HT(2A) and 5-HT(2C) in both hypothalamus and midbrain. In contrast, the only alteration in the hippocampus was 5-HT(6) overexpression. Neither 5-HT transporter nor synthetic enzyme gene transcription differed between housing conditions. In conclusion, early social isolation in mice induces robust changes in postsynaptic 5-HT receptors gene transcription, motor hyperactivity and behavioral disinhibition. The overall pattern of decreased gene expression in the prefrontal cortex highlights its high vulnerability to environment. Furthermore, this is the first study to present a general representation of 5-HT related gene expression in specific brain areas after social isolation and identifies novel candidates that may be critical for underlying molecular mechanisms. PMID- 17083333 TI - Alcoholism is associated with GALR3 but not two other galanin receptor genes. AB - The neuropeptide galanin is widely expressed in the periphery and the central nervous system and mediates diverse physiological processes and behaviors including alcohol abuse, depression and anxiety. Four genes encoding galanin and its receptors have been identified (GAL, GALR1, GALR2 and GALR3). Recently we found that GAL haplotypes were associated with alcoholism, raising the possibility that genetic variation in GALR1, GALR2 and GALR3 might also alter alcoholism risk. Tag single nucleotide polymorphisms (SNPs) were identified by genotyping SNP panels in controls from five populations. For the association study with alcoholism, six GALR1, four GALR2 and four GALR3 SNPs were genotyped in a large cohort of Finnish alcoholics and non-alcoholics. GALR3 showed a significant association with alcoholism that was driven by one SNP (rs3,091,367). Moreover, the combination of the GALR3 rs3,091,367 risk allele and GAL risk haplotypes led to a modestly increased odds ratio (OR) for alcoholism (2.4) as compared with the effect of either GAL (1.9) or GALR3 alone (1.4). Likewise, the combination of the GALR3 and GAL risk diplotypes led to an increased OR for alcoholism (4.6) as compared with the effect of either GAL (2.0) or GALR3 alone (1.6). There was no effect of GALR1 or GALR2 on alcoholism risk. This evidence suggests that GALR3 mediates the alcoholism-related actions of galanin. PMID- 17083334 TI - Seroprevalence of herpes simplex virus type 2 in multiple sclerosis. AB - BACKGROUND: It has been proposed that multiple sclerosis (MS) might be a sexually transmitted disorder. There is evidence that seropositivity to herpes simplex virus type 2 (HSV-2) correlates well with the number of sexual partners. Accordingly, a raised overall HSV-2 seroprevalence in MS would lend support to this theory. MATERIALS AND METHODS: Serum from 497 UK subjects with clinically definite MS was tested for antibodies to HSV-2 and compared with matched historical controls from within and outside London, blood donors and genito urinary medicine (GUM) clinics. RESULTS: The unadjusted MS seropositivity rate was 14%. HSV-2 seroprevalence in MS patients aged 35-64 years was significantly higher overall compared with a non-London general population in an unadjusted comparison. HSV-2 seroprevalence in London MS patients compared with London blood donors was significantly greater irrespective of age, but the MS seropositive rate was lower than GUM clinic attenders. In a logistic regression analysis, increased age, female sex and MS diagnosis all independently increased the odds of seropositivity after adjustment for each other. CONCLUSION: It is concluded that there is increased likelihood of HSV-2 exposure in patients with MS and this may indicate a higher than average number of partners. PMID- 17083335 TI - Interferon-beta1b treatment modulates cytokines in patients with primary progressive multiple sclerosis. AB - OBJECTIVES: It is unknown whether the immunological effects of beta-interferon (IFN-beta) differ in primary progressive multiple sclerosis (PPMS) when compared with relapsing-remitting multiple sclerosis (RRMS). Therefore, we investigated the effects of IFN-beta1b treatment in PPMS on proliferation and cytokine pattern of peripheral blood mononuclear cells (PBMC) and interleukin-10 (IL-10) serum level. METHODS: Eighteen patients were treated with IFN-beta1b for 12 months in an open-label trial. Serum and PBMC were collected longitudinally. RESULTS: Interleukin-10 serum levels increased (P = 0.02) during treatment. Tumor necrosis factor-alpha was increased in anti CD3 (OKT3) antibody stimulated PBMC during treatment (P = 0.04), whereas secretion of IL-10 was decreased in OKT3 (P = 0.04), but increased in concavalin A stimulated PBMC (P = 0.02). CONCLUSIONS: Interleukin-10 serum levels rose in IFN-beta1b-treated patients as has been observed in RRMS. The changes in cytokine patterns secreted by T-lymphocytes of PPMS patients, however, differ from effects observed in RRMS supporting the hypothesis that PPMS differs in some immunological aspects from RRMS. PMID- 17083336 TI - Angiotensin-converting enzyme I/D gene polymorphism and risk of multiple sclerosis. AB - OBJECTIVES: Angiotensin-converting enzyme (ACE) activity is increased in blood and cerebrospinal fluid of patients with multiple sclerosis (MS). In addition, in experimental autoimmune encephalomyelitis (EAE), an animal model of MS, the blockade of ACE suppresses the disease itself. To analyze the genetic association of the ACE gene with MS, we examined ACE gene insertion/deletion (I/D) polymorphism in MS patients. MATERIALS AND METHODS: A total of 313 MS patients from Slovenia and Croatia and 376 healthy controls were genotyped by polymerase chain reaction method. RESULTS: We found statistically significant differences in the distribution of ACE I/D allele frequencies (P < 0.01) and genotypes (P < 0.04) in male patients. ACE DD genotype was associated with MS in men at an odds ratio of 1.86 (95% CI 1.09-3.19, P = 0.02). CONCLUSIONS: DD genotype of ACE gene might contribute to a higher risk of developing MS in men. PMID- 17083337 TI - Stroke risk factor, pattern and outcome in patients with cancer. AB - OBJECTIVE: To investigate risk factors, stroke pattern and outcome in stroke patients with cancer. METHODS: A retrospective review of all stroke patients with cancer (n = 69) admitted to Bankstown-Lidcombe Hospital, Sydney, Australia, between January 1999 and December 2004 was conducted, and comparison made to age- and gender-matched non-cancer stroke patients admitted to the same hospital during the same period. RESULTS: Vascular risk factors did not significantly differ between cancer and non-cancer groups. There was a trend towards greater risk of intracerebral haemorrhage in the cancer group vs the non-cancer group. Previous deep vein thrombosis was more common, and prothrombin time and activated partial thromboplastin time were prolonged in the cancer group vs the non-cancer group. More patients died in hospital in the cancer group vs the non-cancer group. CONCLUSIONS: Coagulation disorders are more likely to be seen in stroke patients with cancer, and patients with cancer have a higher in-hospital post stroke mortality. PMID- 17083338 TI - Peripheral neuropathy in type 2 diabetes mellitus in Isfahan, Iran: prevalence and risk factors. AB - BACKGROUND: To estimate the prevalence and risk factors of peripheral neuropathy (PN) in people with type 2 diabetes mellitus. METHODS: A total of 810 patients with type 2 diabetes (289 men and 521 women) from Isfahan Endocrinology and Metabolism Research Centre outpatient clinics, Iran, were examined. Part of examination included an assessment of neurological function including neuropathic symptoms and physical signs and nerve conduction velocity. RESULTS: The prevalence of PN was 75.1% (95% confidence interval 72.1, 78.0). PN was associated with age, proteinuria, and duration of diabetes, insulin-treatment, and presence of any retinopathy and ischaemic heart disease (IHD). The age adjusted prevalence rate of PN was 78% higher among patients with IHD, 64% higher among patients with any retinopathy, 66% higher among insulin-treated type 2 diabetes, and it was greater with duration of diabetes. Using a stepwise binary logistic regression model, age, duration of diabetes and proteinuria were significant independent predictors of PN. CONCLUSION: Peripheral neuropathy is a common complication in this population of Iranian type 2 diabetic patients. It increases with age, duration of diabetes and proteinuria. PMID- 17083339 TI - Neck pain and associated head pain: persistent neck complaint with subsequent, transient, posterior headache. AB - BACKGROUND: During the Vaga headache epidemiology study, there were indications that various types of work, such as carpentry and hairdressing, frequently seemed to be linked to a combination of neckache and headache. OBJECTIVES: A post hoc study was conducted among 1838, 18- to 65-year-old Vaga citizens, looking for patients with combined neck/headache. DESIGN: Face-to-face interview was conducted. RESULTS: Combined neckache and headache were detected in 121 (6.6%) individuals. The bilateral headache originated in the neck; it was mild/moderate, symptom-poor, and frequently provoked by awkward neck positions. No such headache occurred without a neckache. Headache in this group of patients resembles tractor drivers' headache, except for the provoking factor itself. CONCLUSIONS: This headache is not listed in headache classification systems/textbooks on headache. This headache must be distinguished from the unilateral cervicogenic headache proper because of the different treatment perspectives. PMID- 17083340 TI - Examination of the SLITRK1 gene in Caucasian patients with Tourette syndrome. AB - OBJECTIVE: To determine whether variants in the Slit and Trk-like 1 gene (SLITRK1) are present in American Caucasian population of patients with Tourette syndrome (TS). METHODS: We sequenced the 3'-untranslated region for var321 and the whole coding region in the SLITRK1 gene in 82 Caucasian patients with TS from North America. RESULTS: None of the 82 samples from patients with TS showed the non-coding sequence variant (var321). Only one patient with familial TS was heterozygous for a novel 708C > T (Ile236Ile) nucleotide variant. CONCLUSIONS: The var321 and mutation(s) in the coding region of the SLITRK1 gene probably are a rare cause of TS in a Caucasian population; therefore, genetic heterogeneity of TS should be considered. Tests designed to detect variant(s) in the SLITRK1 gene probably will not have a diagnostic utility in clinical practice. PMID- 17083341 TI - Symptomatic Charcot-Marie-Tooth? A pair of concordant monozygotic twins. AB - BACKGROUND: A pair of monozygotic twin brothers were referred due to hereditary peripheral neuropathy resembling late onset Charcot-Marie-Tooth (CMT). AIM OF THE STUDY: Diagnostic classification of the twin pair. METHOD: Clinical, neurological, genetical and neurophysiological examination, and molecular genetic testing. RESULTS: The clinic and neurophysiology was compatible with CMT disease with late onset. Molecular genetic analysis excluded mutations in PMP22, connexin32, MPZ, LITAF and MFNZ genes, as well as duplication and deletion of PMP22. CONCLUSIONS: The twins were employed in PVC production and developed symptoms after 14 years of massive exposure. We think that the heavy exposure to various neurotoxic compounds caused symptoms that mimic late-onset CMT. However, the twins had distal dysesthesia which is unusual in inherited neuropathies. This illustrates the importance of an occupational history even in the molecular genetic era. PMID- 17083342 TI - Due to low infection rates no routine anti-infective prophylaxis is required in younger patients with chronic lymphocytic leukaemia during fludarabine-based first line therapy. AB - The impact of the combination therapy fludarabine plus cyclophosphamide (FC) in comparison with fludarabine alone regarding the incidence and severity of infections among previously untreated patients with chronic lymphocytic leukaemia (CLL) was evaluated within a multicentre phase III study. A total of 375 patients, up to 65 years old, were randomised between fludarabine or FC for first line therapy. No routine anti-infective prophylaxis was provided. A total of 196 infectious episodes, including 33 severe infections, were documented. In the fludarabine arm, 32.9% of the patients developed an infectious complication compared with 39.9% in the FC arm (P = 0.2). No difference was observed in the rate of severe infections (Common Toxicity Criteria grades III and IV) between both treatment arms. Dose reductions were performed more frequently in FC-treated patients. Granulocyte colony-stimulating factor (G-CSF) was administered due to leucopenia in 5% of all patients. A multivariate regression model identified only elevated thymidine kinase, but not the treatment arm, as a statistically independent risk factor for infections. In summary, FC was not associated with a higher rate of infections compared with fludarabine alone. No routine antibiotic or virostatic prophylaxis, or preemptive treatment with G-CSF, is necessary in first line therapy with fludarabine-based regimens in younger patients with CLL, if adequate dose reduction is performed. The combination therapy FC is not associated with a higher rate of infections compared with fludarabine alone. No routine antibiotic or virostatic prophylaxis as well as preemptive treatment with G-CSF is necessary in first line therapy with fludarabine-based regimen in younger patients with CLL, if adequate dose reductions due to cytopenia or previous infections are performed. PMID- 17083343 TI - Using twin studies to determine genetic and environmental components of allergy and asthma. PMID- 17083344 TI - A need for circulating biomarkers of severe persistent asthma and its treatment. PMID- 17083345 TI - Regulation of the T cell response. AB - The T cell branch of the immune system can respond to a virtually infinite variety of exogenous antigens, thus including the possibility of self-antigen recognition and dangerous autoimmune reactions. Therefore, regulatory mechanisms operate both during ontogeny within the thymus and after birth in the periphery. The control of self-reactive T cells occurs through a process of negative selection that results in apoptosis of T cells showing high affinity for self peptides expressed at the thymic level by means of promiscuous gene expression. Self-reactive T cells escaped to negative selection are controlled in the periphery by other regulatory mechanisms, the most important being natural Foxp3+ T regulatory (Treg) cells. Regulation is also required to control excessive effector T cell responses against exogenous antigens, when they become dangerous for the body. Three types of effector T cells have been recognized: T helper 1 (Th1) cells, which are protective against intracellular bacteria; Th2 cells, which play some role in the protection against nematodes, but are responsible for allergic reactions; Th17 cells, which are probably effective in the protection against extracellular bacteria, but also play a role in the amplification of autoimmune disorders. Abnormal or excessive Th effector responses are regulated by different mechanisms. Redirection or immune deviation of Th1- or Th2-dominated responses is provided by cytokines [interferon-gamma (IFN-gamma) vs. interleukin 4 (IL-4)] produced by the same cell types and by the CXCR3-binding chemokines CXCL4 and CXCL10. Moreover, both Th1 and Th2 responses can be suppressed by adaptive Treg cells through contact-dependent mechanisms and/or the production of IL-10 and transforming growth factor-beta (TGF-beta). Finally, TGF-beta1 can promote the development of both Th17 effector and adaptive Treg cells, while the contemporaneous production of IL-6 contributes to the development of Th17 cells, but inhibits Treg cells. The development of Th17 cells is also down-regulated by IL-4 produced by Th2 cells and by IFN-gamma produced by Th1 cells. PMID- 17083346 TI - Local mucosal immunoglobulin E production: does allergy exist in non-allergic rhinitis? AB - In this review, we critically evaluate the evidence for local IgE production in allergic rhinitis mucosa and the concept of local allergy in non-atopic idiopathic rhinitis. Significantly, fewer studies have focused on the disease pathways associated with non-allergic rhinitis compared with their allergic counterparts. Recently, there's been a revival of the hypothesis concerning the existence of local tissue-specific allergic disease confined to the nasal mucosa of some systemically non-atopic rhinitis subjects. Providing the evidence for local mucosal IgE production in allergic rhinitis is a pre-requisite to reviewing its existence in non-allergic rhinitis. In addition, practical and theoretical approaches useful in the detection of allergy in non-allergic rhinitis will be discussed. Furthermore, successful therapeutic regimens used in the treatment of non-allergic rhinitis will be examined as these could provide an insight into the underlying pathophysiology of this common but poorly understood disease. PMID- 17083347 TI - High serum levels of tumour necrosis factor-alpha and interleukin-8 in severe asthma: markers of systemic inflammation? AB - BACKGROUND: Severe asthma is characterized by elevated levels of pro-inflammatory cytokines and neutrophilic inflammation in the airways. Blood cytokines, markers of 'systemic' inflammation, may be a feature of amplified inflammation in severe asthma. OBJECTIVE: To detect differences in IL-8, TNF-alpha, IL-16 and IL-13 levels in the serum(s) of stable severe and mild-moderate asthmatics related to blood leucocytes proportion, airway calibre and exhaled nitric oxide (NO) levels. METHODS: We assessed cytokine serum levels by ELISA and blood leucocyte counts by an alkaline peroxidase method in 20 healthy controls, 22 mild-moderate [forced expiratory volume in 1 s (FEV1)(%pred): 89+/-3] and 14 severe asthmatics [FEV1(%pred): 49+/-2]. RESULTS: IL-8 and TNF-alpha levels were higher in severe asthmatics than in mild-moderate asthmatics or in controls (P<0.05). No differences in IL-16 and IL-13 levels were detected. Severe asthmatics showed higher circulating neutrophil and eosinophil number than controls (P<0.05). In severe asthmatics, exhaled NO levels were superior than in controls (P<0.05), but inferior than in mild-moderate asthmatics (P<0.05). We found positive correlation between TNF-alpha levels and exhaled NO (r=0.67; P=0.01) or circulating neutrophil counts (r=0.57; P=0.03) in severe asthmatics. CONCLUSION: sTNF-alpha and sIL-8 are markers of 'systemic' inflammation in severe asthmatics, in conjunction with augmented circulating neutrophils, suggesting the involvement of neutrophil-derived cytokine pattern in severe asthma. PMID- 17083348 TI - Multivariate genetic analysis of atopy phenotypes in a selected sample of twins. AB - BACKGROUND: Atopic traits often co-occur and this can potentially be caused by common aetiological relationships between traits, i.e. a common genetic or a common environmental background. OBJECTIVE: To estimate to what extent the same genetic and environmental factors influence wheeze, rhinitis, airway hyper responsiveness (AHR), and positive skin prick test (posSPT) in a sample of adult twins. METHODS: Within a sampling frame of 21,162 twin subjects, 20-49 years of age, from the Danish Twin Registry, a total of 575 subjects (256 intact pairs and 63 single twins), who either themselves and/or their co-twins reported a history of asthma at a nationwide questionnaire survey, were clinically examined. Symptoms of wheeze and rhinitis were obtained by interview; airway responsiveness and skin test reactivity were measured using standard techniques. Correlations in liability between the different traits were estimated and latent factor models of genetic and environmental effects were fitted to the observed data using maximum likelihood methods. RESULTS: The various phenotypic correlations between wheeze, rhinitis, AHR and posSPT were all significant and ranged between 0.50 and 0.86. Traits that showed highest genetic correlations were wheeze-rhinitis (rho(A)=0.95), wheeze-AHR (rho(A)=0.85) and rhinitis-posSPT (rho(A)=0.92), whereas lower genetic correlations were observed for rhinitis-AHR (rho(A)=0.43) and AHR-posSPT (rho(A)=0.59). Traits with a high degree of environmental sharing were rhinitis-posSPT (rho(E)=0.92) and wheeze-posSPT (rho(E)=0.71), whereas a lower environmental correlation was seen for wheeze-rhinitis (rho(E)=0.25). The estimates were corrected for ascertainment and adjusted for age, sex, inhaled corticosteroids and smoking. CONCLUSIONS: Different atopic conditions share, to a large extent, a common genetic background. In particular, upper and lower respiratory symptoms seem to be different phenotypic expressions of a common set of genes. These results add new insight into the origins of clinical heterogeneity within atopy and should stimulate the search for pleiotropic genes of importance for these conditions. PMID- 17083349 TI - Interaction between variants in the interleukin-4 receptor alpha and interleukin 9 receptor genes in childhood wheezing: evidence from a birth cohort study. AB - BACKGROUND: Several polymorphisms in the IL-4 receptor alpha (IL4RA) gene have been associated with asthma and atopy, but with variable success in different populations. Immunologic studies suggest that IL4RA may interact with other cytokines and receptors, and gene-gene interactions have also been observed with respect to asthma. Such interactions have been proposed to explain partly the difficulties in replicating association studies. METHODS: Using the prospective birth cohort BAMSE, we examined eight single nucleotide polymorphisms (SNPs) and corresponding haplotypes in the IL4RA gene in relation to wheezing and sensitization up to age 4. We also evaluated potential interaction effects (departure from a multiplicative interaction model) between the IL4RA SNPs and four SNPs in the IL-9 receptor (IL9R) gene previously associated with childhood wheezing. RESULTS: We found no main effect of the IL4RA SNPs alone and only weak associations to wheezing and sensitization when haplotypes were considered. Gene gene interactions between several IL4RA and IL9R SNPs with regard to wheezing were observed (P=0.009), especially between IL4RA Q576R (rs1801275) and IL9R rs731476 (P=0.005). An interaction was also seen between IL4RA and IL9R haplotypes. CONCLUSION: Variants in the IL4RA gene alone may not exert any major influence on susceptibility to asthma-related diseases in childhood, but in combination with other genes, such as IL9R, IL4RA may be an important gene for disease susceptibility. PMID- 17083350 TI - Genetic selection for resistance or susceptibility to oral tolerance imparts correlation to both Immunoglobulin E level and mast cell number phenotypes with a profound impact on the atopic potential of the individual. AB - BACKGROUND: Immunological oral tolerance is being studied with great interest due to its therapeutic potential in allergy and autoimmunity processes, although the cellular and molecular mechanisms linking these different phenomena remain elusive. In the present study, two mouse lines with extreme phenotypes for susceptibility [TS Line] or resistance [TR Line] to oral tolerance and their [TS x TR]F2 segregants were used in order to evaluate the impact of these traits on the atopic potential of the individuals. OBJECTIVE: Demonstrate whether the tr and ts genes, cumulated during 18 generations of bidirectional genetic selection, influence expression of two important immunobiological traits (IgE and mast cell) critical to allergic response. METHODS: Mice with extreme phenotypes for oral tolerance to ovalbumin (OVA), produced by assortative mating (TS and TR Line), and their (TS x TR)F2 segregating were used. Serum IgE levels assayed by ELISA, and mastocytes counted with toluidine blue staining were evaluated in naive mice. Anaphylaxis was induced by intravenous injection of OVA, intestinal inflammation by oral administration of OVA 7 days after immunization, and pulmonary inflammation by intranasal and nebulization OVA challenges. Specific IgE was dosed by passive cutaneous anaphylaxis. RESULTS: The naive TS mice have a 20-fold lower serum IgE level and two- to threefold diminished mast cell numbers in mucosal sites, when compared with TR-mice, which were highly susceptible to allergic inflammation and anaphylactic shock. The associations of oral tolerance, serum IgE levels and mast cell numbers in naive animals were confirmed analysing the simultaneous presence of these traits in individuals of a [TS x TR]F2 segregating population. CONCLUSION: The results suggest that the complex of genes controlling TS and TR phenotypes play a main role in the regulation of the atopic potential of the individual. The studies of these traits in interline F2 segregants demonstrated a co-segregation of TS and TR phenotypes with IgE responsiveness and mast cell numbers. Thus, the opposite capacity of the genetically modified mice may be involved in co-adaptative mechanisms reflecting a dynamic relation between gene frequencies in a natural population. These correlations give circumstantial evidence to support clinical applications of oral tolerance in allergic and autoimmune diseases. PMID- 17083351 TI - Association of neuropeptides with Th1/Th2 balance and allergic sensitization in children. AB - BACKGROUND: Among neurogenic factors, the neuropeptides have an important regulatory influence on immune system activity and may lead to allergic sensitization. OBJECTIVE: The aim of our study was to investigate the relationship of the neuropeptides vasoactive intestinal peptide (VIP), somatostatin (SOM) and substance P (SP) on modulation of Th1/Th2 balance and allergic sensitization in children. METHODS: Within the LISAplus (Life style Immune system-Allergy) study, blood samples of 321 six-year-old children were analysed for concentration of neuropeptides, Th1 and Th2 cytokines, transcription factors for T cell regulation and suppressors of cytokine signalling. In addition, samples were screened for specific IgE against inhalant and food allergens. RESULTS: Children with high SOM values showed a Th2 polarization and a reduced expression of FOXP3, the marker for regulatory T cells. High (VIP) levels correlated inversely with the expression of T cell transcription factors (Tbet and SOCS3). In contrast, elevated levels of SP were associated with reduced GATA3 and SOCS3 expression and with increased IFN-gamma concentrations. Allergic sensitization was more prevalent in children with higher SOM and VIP concentrations but not associated with SP levels. CONCLUSION: Our data reveal an association between neuropeptides and modulatory effects on immune cells in vivo, especially on Th1/Th2 balance with a correlation to allergic sensitization in children. We suggest that elevated SOM and VIP concentrations and the inducing factors should be considered as allergy risk factors. PMID- 17083352 TI - Effects of inhaled ciclesonide on circulating T-helper type 1/T-helper type 2 cells in atopic asthmatics after allergen challenge. AB - BACKGROUND: The predominance of T-helper type 2 (Th2) lymphocytes is thought to underlie the pathogenesis of asthma. Allergen inhalation challenge in atopic asthmatic subjects is associated with decreased interferon-gamma (IFN-gamma) positive CD4+ and CD8+ lymphocytes in peripheral blood and induced sputum. OBJECTIVE: This study examined the effects of an inhaled corticosteroid on these previously described allergen-induced changes in circulating Th1 and Th2 lymphocytes. METHODS: Subjects were randomized to 7 days of placebo, 40 or 80 micro g ciclesonide in a crossover study. Airway responses and peripheral blood were measured before and after treatment, and 24 h after allergen challenge. RESULTS: Ciclesonide 40 and 80 micro g significantly attenuated the late response and sputum eosinophils at 8 h post-allergen (P<0.05). Circulating IFN-gamma positive CD4+ lymphocytes decreased after allergen challenge with placebo (P<0.05), and this was inhibited by 40 micro g ciclesonide treatment (P<0.05). There was no effect of allergen inhalation or ciclesonide on IL-4-positive CD4+ lymphocytes or IFN-gamma and IL-4-positive CD8(high) lymphocytes. The allergen induced change of IFN-gamma/IL-4 ratio on CD4+ cells correlated with the allergen induced change of peripheral blood eosinophils. CONCLUSIONS: The results of this study suggest that attenuation of allergen-induced airway responses by ciclesonide may be mediated through regulation of IFN-gamma-positive CD4+ cells. PMID- 17083353 TI - Probiotics in the treatment of Japanese cedar pollinosis: a double-blind placebo controlled trial. AB - BACKGROUND: Probiotic bacteria may be effective in the treatment of allergic inflammation and food allergy, but efficacy and underlying mechanisms remain unclear. OBJECTIVE: The present study investigated the effects of probiotic strain Bifidobacterium longum BB536 in the treatment of Japanese cedar pollinosis (JCPsis). METHODS: In a randomized, double-blind, placebo-controlled trial, 44 JCPsis subjects received BB536 or placebo for 13 weeks during the pollen season. Subjective symptoms and self-care measures were recorded daily and blood samples were taken before and during intervention to measure blood levels of parameters related to JCPsis. RESULTS: BB536 intake was associated with a significant reduction in number of subjects prematurely terminated due to severe symptoms and pollinosis medication (P=0.0057 vs. placebo group). Comparison of subjective symptom scores indicated significant decreases in rhinorrhea, nasal blockage and composite scores in the BB536 group compared with the placebo group. Comparison of medical scores showed marked improvements in all symptoms on BB536 intake. A T helper type 2 (Th2)-skewed immune response occurring along with pollen dispersion was observed. BB536 significantly suppressed increases in plasma thymus- and activation-regulated chemokine and tended to suppress elevations of Japanese cedar pollen (JCP)-specific IgE. CONCLUSION: These results suggest the efficacy of BB536 in relieving JCPsis symptoms, probably through the modulation of Th2 skewed immune response. PMID- 17083354 TI - CD2 identifies a monocyte subpopulation with immunoglobulin E-dependent, high level expression of Fc epsilon RI. AB - BACKGROUND: Fc epsilon RI expression by monocytes can affect monocyte function via multiple mechanisms, thereby potentially influencing the generation of allergic inflammation. Previous studies on the in vivo regulation of monocyte Fc epsilon RI expression by ambient IgE have yielded conflicting results. OBJECTIVE: We hypothesized that monocyte Fc epsilon RI expression is limited to a specific monocyte subset, and that within that subset Fc epsilon RI surface expression is correlated to serum IgE. METHODS: Study 1: Blood was obtained from non-allergic subjects (n=14) and subjects with allergic asthma (n=18), hypereosinophilic syndrome (n=2), hyper-IgE syndrome (n=6), and helminth infection (n=4). Study 2: Blood was obtained from allergic subjects in a clinical trial of omalizumab before and during study drug treatment. Monocyte surface Fc epsilon RI expression was measured using flow cytometry. RESULTS: Fc epsilon RI expression was significantly greater in the CD2(high) vs. CD2(low) monocyte subsets (31% vs. 1.9% median Fc epsilon RI+, respectively). In asthmatic and non-atopic healthy control subjects, CD2(low) monocytes expressed little or undetectable Fc epsilon RI. In study 1, Fc epsilon RI expression was highly correlated to serum IgE in the CD2(high), but not in the CD2(low) monocyte subpopulation (R values of 0.67 and 0.41, respectively). In study 2, omalizumab, but not placebo, caused a significant and sustained decline in Fc epsilon RI expression within the CD2(high) monocyte subset. CONCLUSIONS: CD2 defines a monocyte subset with high Fc epsilon RI expression, whose magnitude is highly correlated to serum IgE. As such, this new description of CD2(high) monocytes as Fc epsilon RI-bearing cells suggests that they may be potential targets of anti-IgE immunomodulatory therapies. PMID- 17083355 TI - Natural rubber latex and hymenoptera venoms share ImmunoglobinE-epitopes accounting for cross-reactive carbohydrate determinants. AB - BACKGROUND: Epidemiological data on the prevalence and risk factors of latex sensitization have suggested a significant association between latex sensitization and the presence of one or more positive skin prick test responses to aeroallergens, food allergens and to one or more insect venoms. Xylose and core 3-fucose are typical complex glycans in plants and are foreign to mammals. Plant N-glycans and insect N-glycans may cross-react in humans. OBJECTIVE: The aim of our study was to investigate whether there are cross-reactive IgE-binding structures in natural rubber latex (NRL) and hymenoptera venoms and to examine their nature. METHODS: Hundred and twenty-five consecutive patients with insect venom allergy were screened for coincidental latex-specific IgE. IgE-binding components in the venoms from Apis mellifera and/or vespula species and in NRL extracts were characterized by IgE-immunoblotting to the natural allergen sources and determination of specific IgE to recombinant allergens. Cross-reactive components were investigated by inhibition experiments. The involvement of carbohydrates in the constitution of cross-reactive IgE-epitopes was further examined by specific IgE-binding to cross-reactive carbohydrate determinants (CCD) in bromelain and horseradish peroxidase as well as by periodate treatment. RESULTS: NRL glove extracts inhibited patients' serum IgE-binding to venom allergens. Vice versa, the IgE-binding to latex glove extracts could be inhibited by pre-incubation with the insect venoms. Specific IgE-binding to recombinant latex allergens was absent, whereas the cross-reactive IgE-epitopes were sensitive to periodate treatment and specific IgE to CCD (MMXF and MUXF type) could be detected. CONCLUSION: Insect venoms and NRL share IgE-binding CCD that may be responsible for positive serological test results to NRL in patients with insect venom allergy. This copositivity occurs frequently (13.6%) among venom allergic individuals and did not elicit clinical symptoms upon contact to latex in the patients examined. In contrast, true cosensitization to insect venoms and NRL allergens can occur and may not be missed. PMID- 17083356 TI - Do shrimp-allergic individuals tolerate shrimp-derived glucosamine? AB - BACKGROUND: There is concern that shrimp-allergic individuals may react to glucosamine-containing products as shrimp shells are a major source of glucosamine used for human consumption. OBJECTIVE: The purpose of this study was to determine whether shrimp-allergic individuals can tolerate therapeutic doses of glucosamine. METHODS: Subjects with a history of shrimp allergy were recruited and tested for both shrimp reactivity via a prick skin test and shrimp-specific IgE by an ImmunoCAP assay. Fifteen subjects with positive skin tests to shrimp and an ImmunoCAP class level of two or greater were selected for a double-blind placebo-controlled food challenge (DBPCFC) using glucosamine-chondroitin tablets containing 1,500 mg of synthetically produced (control) or shrimp-derived glucosamine. Immediate reactions, including changes in peak flow and blood pressure, and delayed reactions (up to 24 h post-challenge) via questionnaire were noted and assessed. RESULTS: All subjects tolerated 1,500 mg of both shrimp derived or synthetic glucosamine without incident of an immediate hypersensitivity response. Peak flows and blood pressures remained constant, and no subject had symptoms of a delayed reaction 24 h later. CONCLUSION: This study demonstrates that glucosamine supplements from specific manufacturers do not contain clinically relevant levels of shrimp allergen and therefore appear to pose no threat to shrimp-allergic individuals. PMID- 17083357 TI - Effects of phthalate esters on the sensitization phase of contact hypersensitivity induced by fluorescein isothiocyanate. AB - BACKGROUND: Many different types of phthalate ester are used as plasticizers and are thus found in the air. There have been several studies that suggest an association between allergies and phthalate esters. We previously found that di butyl phthalate (DBP) has an adjuvant effect in a mouse contact hypersensitivity model, in which fluorescein isothiocyanate (FITC) is involved as an immunogenic hapten. OBJECTIVE: We examined whether other phthalate esters enhance the process of sensitization to FITC by facilitating the trafficking of FITC-presenting dendritic cells or macrophages from skin sites to draining lymph nodes. METHODS: Mice were epicutaneously sensitized with FITC dissolved in acetone containing a phthalate ester. Sensitization was evaluated as ear swelling after a challenge with FITC. Draining lymph node cells obtained 24 h after skin sensitization were examined for FITC fluorescence by means of flow cytometry. FITC-positive cells were characterized with anti-CD11c and anti-CD11b by three-colour flow cytometry. RESULTS: When mice were sensitized with FITC in acetone containing DBP or di-n propyl phthalate (DPP), strong enhancement of the ear-swelling response was observed. Di-methyl phthalate (DMP) and di-ethyl phthalate (DEP) were less effective but produced some enhancement. Consistent enhancement was not observed with di-(2-ethylhexyl) phthalate or di-isononyl phthalate. Upon sensitization in the presence of DBP or DPP, the number of FITC-positive dendritic cells (total CD11c+ as well as CD11c+/CD11b+) was increased in draining lymph nodes. As to the other four phthalate esters, there was no significant increase in the FITC positive cell number in the draining lymph nodes. CONCLUSION: During the process of sensitization to FITC, DBP, and DPP exert strong adjuvant effects that are associated with enhancement of trafficking of antigen-presenting dendritic cells from the skin to draining lymph nodes. PMID- 17083358 TI - Ultrafine but not fine particulate matter causes airway inflammation and allergic airway sensitization to co-administered antigen in mice. AB - BACKGROUND: Airborne particulate matter (PM) is an important factor associated with the enhanced prevalence of respiratory allergy. The PM adjuvant activity on allergic sensitization is a possible mechanism of action involved, and the induction of airway inflammation is suggested to be of importance in PM-induced adjuvant activity. OBJECTIVE: Because differently sized PM have different toxic potentials, we studied the role of particle size in the induction of airway inflammation and allergic sensitization. This was done using fine (0.250 and 0.260 micro m) and ultrafine (0.029 and 0.014 micro m) titanium dioxide (TiO(2)) and carbon black particles (CBP) with known differences in airway toxicity. METHODS: Mice were intranasally exposed to ovalbumin (OVA) alone or in combination with one of the different particles. The induction of airway inflammation and the immune adjuvant activity were studied in the lungs and lung draining peribronchial lymph nodes (PBLN) at day 8. OVA-specific antibodies were measured at day 21, and the development of allergic airway inflammation was studied after OVA challenges (day 28). RESULTS: When administered at the same total particle mass (200 micro g), exposure to ultrafine TiO(2) and CBP-induced airway inflammation, and had immune adjuvant activity. The latter was shown by increasing both the PBLN cell numbers and the production of OVA-specific T-helper type 2 (Th2) cytokines (IL-4, IL-5, IL-10 and IL-13). Whereas OVA-specific IgE and IgG1 levels in serum were only increased in animals exposed to the ultrafine TiO(2), allergic airway inflammation could be detected in both ultrafine TiO(2) and CBP-treated groups after challenges with OVA. CONCLUSION: Our data show that only the ultrafine particles, with a small diameter and a large total surface area/mass, cause airway inflammation and have immune adjuvant activity in the current model supporting the hypothesis that particle toxicity is site-dependent and related to adjuvant activity. PMID- 17083360 TI - Biology of the wool follicle: an excursion into a unique tissue interaction system waiting to be re-discovered. AB - Wool fibres are hairs and the term 'wool' is usually restricted to describe the fine curly hairs that constitute the fleece produced by sheep. In a broader sense, it can be used to describe the fleeces produced by related species such as goat or yak. Research into the biology of wool growth and the structure of the wool fibre has been driven by the demands of the wool industry to improve both the efficiency of growing wool and the quality of the product. Well beyond this very applied perspective however, the wool follicle is a unique basic research model for the life sciences in general. These unique features include, to name just a few selected examples, accessibility for studying the molecular controls involved in branching of secondary epithelial-mesenchymal structures, the photoperiod-dependence of regenerating tissue interaction systems, the origin of fibre curliness and follicle wave pattern formation, and the effect of alterations in nutrient supply on epithelial growth and fibre structure. In this review, investigation of growth processes in the formation of the wool fibre is broadly surveyed. The relevance and potential for practical outcomes through characterization of wool follicle genes are discussed and particular features of the wool follicle contributing to our knowledge of the biology of hair growth are highlighted. The practical potential of gene discovery in wool research is the provision of molecular markers for selective breeding and for altering wool growth and wool structure by other biological pathways such as sheep transgenesis that could lead to novel wool properties. In this background, the current review attempts to revive general interest in the fascinating biology of the wool follicle which is not only of profound economic and practical importance but offers an exquisite, highly instructive research model for addressing key questions of modern biology. PMID- 17083361 TI - Expression pattern of somatostatin receptor subtypes 1-5 in human skin: an immunohistochemical study of healthy subjects and patients with psoriasis or atopic dermatitis. AB - In psoriasis and atopic dermatitis, the inflammatory events have neurogenic components and the neuropeptides modify the functions of immuno-active cells in the skin. Somatostatin is a neuropeptide with several neuroendocrine and immunomodulating properties and mediates its actions by five distinct subtypes of G-protein-coupled receptors (SSTR1-5). This study describes the distribution of SSTR1-5, analysed with immunohistochemistry, in psoriasis, atopic dermatitis and controls. Normal human skin and lesional skin from patients with psoriasis or atopic dermatitis showed many similarities, but also some differences, as regards SSTR expression. SSTR1-3 were strongly expressed in the epidermis of healthy skin, and in the skin of patients with psoriasis or atopic dermatitis. It is noteworthy that SSTR4 and 5 were strongly expressed in the epidermis of psoriasis patients, but weakly expressed in the epidermis of those with atopic dermatitis and normal skin. The intensity of the staining also varied considerably between the different layers of the epidermis, especially in psoriasis patients. In all cases, the dendritic cells, found mostly in the papillary and upper reticular dermis, showed a strong expression of SSTR1-4, but a weak expression of SSTR5. SSTR1-5 were strongly expressed in the sweat glands in all skin biopsies. Hair follicles and sebaceous glands expressed all five subtypes. Striated muscle fibres showed an intense positive expression of SSTR1-4, but a weak or negative expression of SSTR5. The wide distribution and expression pattern of all five SSTRs in human skin suggest that somatostatin is involved in the interactions between the nervous system and the skin. PMID- 17083362 TI - Glutathione S-transferase and CYP1A1 gene polymorphisms and non-melanoma skin cancer risk in Italian transplanted patients. AB - Solid organ transplant recipients are at higher risk of non-melanoma skin cancer (NMSC), especially basal cell carcinoma (BCC) and squamous cell carcinoma (SCC). Genetic alterations in the production of detoxifying enzymes such as glutathione S-transferase (GST) and CYP1A1 may enhance this risk. We investigated the frequency of GST genotypes (GSTM1, GSTM3, GSTT1 and GSTP1) and CYP1A1 in 239 transplant recipients: 107 cases with NMSC and 132 controls free from NMSC matched for type of transplanted organ, duration of transplantation, sex and age. Allele GSTP1*A was associated with a higher risk of NMSC [odds ratio (OR) 1.7 (1.1-2.5); P = 0.017]. Homozygosity for allele GSTP1 Val(105) was lower in cases [OR 0.3 (0.1-0.8); P = 0.012], especially in patients with SCC [OR 0.1 (0.0-0.7); P = 0.012]. A higher risk of BCC was found in patients with GSTM1 null/null [null/null versus A + B, OR 3.1 (1.4-6.8); P = 0.003]. Analysis of allelism and interaction between allelic variants showed significant association between combined GSTM1 and CYP1A1 Val(462) genotypes, where individuals homozygous for the risk allele GSTM1 null and carrying also the allele CYP1A1 Val(462), show a higher risk of developing NMSC [OR 4.5 (1.1-21.4); P = 0.03], especially SCC [OR 6.5 (1.4-34.4); P = 0.01]. GSTP1 polymorphisms are associated with both BCC and SCC risk. GSTM1 polymorphisms seem to be involved in BCC risk, while GSTM1 null/null genotype combined with CYP1A1 allele Val(462) are associated with a higher risk for SCC, indicating that allelism and/or interactions between allelic variants at other loci may also influence the risk of NMSC, particularly SCC. PMID- 17083363 TI - Diverse phenotype of Brooke-Spiegler syndrome associated with a nonsense mutation in the CYLD tumor suppressor gene. AB - Brooke-Spiegler syndrome (BSS) is an autosomal dominant disease characterized by cylindromas, trichoepitheliomas and occasionally spiradenomas. The disease gene was mapped to 16q12-13, and mutations in the CYLD gene were identified in families with BSS. In the present report, we describe a large consanguineous Chinese family with BSS showing an intra-family phenotypic variability. Clinically, some affected individuals only revealed discrete small skin-coloured tumors whereas the proband showed an expansion of multiple large tumors on the back of nose and numerous dome-shaped papules on her scalp. Histologically, both trichoepitheliomas and cylindromas were found in the affected individuals. By sequence analysis, we identified a recurrent mutation 2272C>T (R758X) of the CYLD gene in the affected individuals of this family, which was previously identified in other ethnic families with familial cylindromatosis. Our result provided additional information for phenotype-genotype correlation in BSS. PMID- 17083364 TI - Oestrogen receptor-beta expression in melanocytic lesions. AB - Melanomas rarely occur before puberty, have a higher death rate for males, and tend to be more invasive during pregnancy. Prior to the discovery of a second oestrogen receptor (ERbeta), studies with the initial oestrogen receptor, ERalpha, showed no obvious role for oestrogen in the pathophysiology of benign or malignant melanocytic lesions. To investigate the specific immunostaining patterns of ERalpha and ERbeta, benign nevocytic nevi, dysplastic nevi with mild, moderate and severe cytological atypia, lentigo malignas and melanomas of varying depth (Clark) and thickness (Breslow) were studied. ERbeta but not ERalpha was the predominant oestrogen receptor we found in all types of benign and malignant melanocytic lesions. The most intense ERbeta immunostaining was seen in melanocytes in dysplastic nevi with severe cytological atypia and in lentigo malignas. ERbeta expression levels also correlated with the malignant tumor microenvironment; i.e., melanocytes in proximity with keratinocytes>deeper dermal melanocytes in contact with stroma>minimally invasive melanomas>Clark Level III/IV or thick melanomas (Breslow). Discovery that ERbeta expression varies in relation to the tumor microenvironment and increasing depth of invasion suggests its possible usefulness as a surrogate marker for neoplasia and prognosis in malignant melanoma. PMID- 17083365 TI - Non-steroidal anti-inflammatory drugs selectively inhibit cytokine production by NK cells and gamma delta T cells. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) are known to be risk factors for a systemic inflammatory syndrome in viral infections. Innate immune cells are likely to represent the preferential targets for the deleterious effects of NSAIDs in patients with viral infections. We therefore examined whether various classes of NSAIDs could selectively inhibit cytokine production by innate immune cells. NSAIDs selectively inhibited interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha production by natural killer (NK) and gammadelta T cells with each NSAID displaying its own unique pattern of inhibition, while sparing that by acquired immune cells. These inhibitions were independent on cyclooxygenase inhibition. These NSAIDs directly inhibited the cytokine production by the purified gammadelta T-cell population without involving other cell populations. The selective inhibition of the early generation of IFN-gamma and TNF-alpha from NK and gammadelta T cells by NSAIDs may serve to drive the subsequent acquired immune responses towards a Th2 phenotype, leading to the aggravation of allergic symptoms. Our results provide a mechanism to explain the deleterious effects of NSAIDs on clinical symptoms of viral infections and allergic diseases and suggest more targeted use depending on the type of disease. PMID- 17083366 TI - Interleukin (IL)-19, IL-20 and IL-24 are produced by and act on keratinocytes and are distinct from classical ILs. AB - Due to their structural similarity, interleukin (IL)-19, IL-20, IL-22, IL-24 and IL-26 were combined with IL-10 in the so-called IL-10 family. To expand the knowledge on IL-19, IL-20 and IL-24, we systematically and quantitatively analysed the expression of these mediators and their receptor chains in vitro and in vivo under various conditions and in comparison with other IL-10 family members. In vitro, IL-19, IL-20 and IL-24 were produced not only by activated immune cells, particularly monocytes, but also to a similar extent by keratinocytes. IL-1beta increased the expression of these mediators 1000-fold (IL 19) and 10-fold (IL-20 and IL-24) in keratinocytes. In vivo, these cytokines were expressed preferentially in inflamed tissues. The absence of either R1 chain for the two types of receptor complexes for these cytokines (IL-20R1/IL-20R2 and IL 22R1/IL-20R2) on immune cells implies that they cannot act on these cells. In fact, IL-19, IL-20 and IL-24 did not induce activation of signal transducer and activator of transcription (STAT) molecules in immune cells. Instead, several tissues, particularly the skin, tissues from the reproductive and respiratory systems, and various glands appeared to be the main targets of these mediators. Keratinocytes expressed both receptor complexes; however, the expression of IL 22R1 was 10 times higher than that of IL-20R1. Interferon-gamma further increased the expression of IL-22R1 and decreased that of IL-20R1, suggesting that under T1 cytokine conditions these mediators primarily affect keratinocytes via the IL 22R1/IL-20R2 complex. In summary, these data support the notion that IL-19, IL-20 and IL-24 are distinct from classical ILs and constitute a separate subfamily of mediators within the IL-10 family. PMID- 17083367 TI - 'To repair or not to repair - no longer a question': repair of mitochondrial DNA shielding against age and cancer. AB - The role of mitochondria in energy production and apoptosis is well known. The role of mitochondria and particularly the role of the mitochondria's own genome, mitochondrial (mt) DNA, in the process of ageing were postulated decades ago. However, this was discussed, debated and more or less disposed of. Recent data from elegant mouse models now confirm that mutations of mtDNA do indeed play a central and pivotal role in the ageing process. Newer reports also indicate a possible role of mtDNA mutations in the carcinogenesis of several organs. But is damaged mtDNA repaired, or is it simply degraded and discarded? This question appears to be answered now. According to recent data, mitochondria possess functional repair mechanisms such as base excision repair, double-strand break repair and mismatch repair, yet nucleotide excision repair has so far not been detected. PMID- 17083368 TI - Desmoglein antibodies and pemphigus vulgaris: a tale of treason and a fight for life. PMID- 17083369 TI - Nutrition acquisition strategies during fungal infection of plants. AB - In host-pathogen interactions, efficient pathogen nutrition is a prerequisite for successful colonization and fungal fitness. Filamentous fungi have a remarkable capability to adapt and exploit the external nutrient environment. For phytopathogenic fungi, this asset has developed within the context of host physiology and metabolism. The understanding of nutrient acquisition and pathogen primary metabolism is of great importance in the development of novel disease control strategies. In this review, we discuss the current knowledge on how plant nutrient supplies are utilized by phytopathogenic fungi, and how these activities are controlled. The generation and use of auxotrophic mutants have been elemental to the determination of essential and nonessential nutrient compounds from the plant. Considerable evidence indicates that pathogen entrainment of host metabolism is a widespread phenomenon and can be accomplished by rerouting of the plant's responses. Crucial fungal signalling components for nutrient-sensing pathways as well as their developmental dependency have now been identified, and were shown to operate in a coordinate cross-talk fashion that ensures proper nutrition-related behaviour during the infection process. PMID- 17083370 TI - P-glycoprotein functions as a differentiation switch in antigen presenting cell maturation. AB - P-glycoprotein (P-gp) expressed on human antigen presenting cells (APC) regulates alloantigen-dependent T-cell activation, but the associated mechanisms are not well understood. Here we demonstrate that P-gp functions in IL-12-dependent monocyte differentiation into dendritic cell (DC) lineages during APC maturation, thereby regulating the capacity of myeloid-derived APCs to elicit alloimmune Th1 responses. Human CD14+ monocytes cultured in vitro in the presence of IL-4/GM-CSF differentiated into CD14(-) CD1A+ APCs of the immature DC phenotype. In contrast, P-gp blockade during differentiation inhibited CD1a induction, down-regulated CD80 expression, enhanced CD86 expression and induced CD68 expression. APCs differentiated in the presence of P-gp blockade stimulated alloimmune T-cell proliferation significantly less than controls and this effect was associated with 97% inhibition of Th1 IFN-gamma production, but preserved Th2 IL-5 secretion. MAb-mediated blockade of the P-gp transport substrate IL-12 in the course of APC differentiation also inhibited IFN-gamma production, while addition of rIL-12 to P-gp-blocked APC differentiation cultures significantly reversed this effect, demonstrating that P-gp functions in APC differentiation in part via IL-12 regulation. Our findings define a novel role for P-gp as a differentiation switch in APC maturation and resultant alloimmune Th1 responses, thereby identifying P-gp as a potential novel therapeutic target in allotransplantation. PMID- 17083371 TI - Helicobacter pylori vaccines: is DNA the answer? PMID- 17083372 TI - Development and evaluation of a DNA vaccine based on Helicobacter pylori urease B: failure to prevent experimental infection in the mouse model. AB - BACKGROUND: The development of a vaccine against Helicobacter pylori has become a priority to prevent major morbidity and mortality associated with this infection. Our goal was to prepare and evaluate a DNA vaccine based on the urease B gene (ureB). METHODS: The ureB gene of H. pylori was amplified and cloned into the eukaryotic expression vector pcDNA3.1/TOPO. Plasmid DNA was purified from transformed Escherichia coli cells and used to immunize mice by the intragastric, intramuscular, intrarectal (40 micro g each) and intranasal (16 micro g) route, three doses every 2 weeks, with CpG oligodeoxynucleotide (ODN) as adjuvant. Four weeks after the third dose, animals were orally challenged with Helicobacter felis and were sacrificed 6 weeks later. The stomach was stained to detect the presence of infection. RESULTS: Despite in vitro confirmation of successful cloning and functionality of the ureB gene with expression of a protein morphologically and antigenically identical to urease B, the DNA vaccine did not perform well in vivo. Immunization of mice produced a weak immune response. Overall, intrarectal and intranasal administration seemed more immunogenic than other routes. Protection against challenge was modest and nonsignificant, and slightly better on animals immunized by the intramuscular and intranasal route. CONCLUSION: A DNA vaccine based on H. pylori urease B was poorly immunogenic and nonprotective at the conditions evaluated. Higher doses, better adjuvants or a prime-boost approach may circumvent these limitations. PMID- 17083373 TI - Helicobacter pylori infection: further evidence for the role of feco-oral transmission. AB - BACKGROUND: Helicobacter pylori infection is recognized as a major cause of chronic digestive diseases with a major public health impact, yet the knowledge of transmission pathways is limited. We studied the transmission in employees taking care of institutionalized persons with mental disabilities with a documented high prevalence of H. pylori. MATERIALS AND METHODS: Six hundred and seventy-one health-care workers were screened for H. pylori serology. For each employee, information was collected on age, sex, father's and mother's education level, number of household members and number of children sleeping in the same bedroom during childhood, as well as lifestyle factors such as smoking and tropical journeys and occupational exposure data such as type of contact with inhabitants (changing napkins with stools, washing inhabitants, feeding inhabitants, personal contact) and seniority in the institution. RESULTS: Seroprevalence for H. pylori increased significantly with age. In univariate analysis, risk factors for H. pylori positivity were (age-adjusted): father's education, mean length of employment, smoking, contact with fecal materials of inhabitants, washing and feeding of inhabitants. Controlling for confounders, in multiple logistic regression analysis, only fecal contact remained as a significant risk factor for H. pylori infection. CONCLUSIONS: In health-care workers caring for a population with a high prevalence of H. pylori infection, there is an association with fecal transmission. This, however, does not rule out the possibility of other ways of transmission. PMID- 17083374 TI - Helicobacter felis infection causes an acute iron deficiency in nonpregnant and pregnant mice. AB - BACKGROUND: The role of Helicobacter pylori infection in iron deficiency during pregnancy is limited. The aim of the present study was to assess the relationship between Helicobacter infection and levels of iron stores in pregnant mice. MATERIALS AND METHODS: Female C57BL/6 mice were either inoculated with 10(8) H. pylori, Helicobacter felis or water. In the nonpregnant study, 15 mice from each group were sacrificed after 4 and 20 weeks of infection. In the pregnancy study, after 6 weeks of infection all female mice were mated and approximately 2 weeks after mating, half of the pregnant mice (n = 9/group) from each group were sacrificed. The remaining mice were allowed to give birth, and approximately 4 weeks after birth, mice were asphyxiated with CO2, followed by heart puncture, and killed by cervical dislocation. Serum ferritin and iron were determined with a micro-particle enzyme immunoassay method and by a timed-endpoint method. RESULTS: Serum iron levels in mice infected with H. felis were significantly (p < .05) lowered compared to control (24%) and H. pylori (27%)-infected mice at 4 weeks of infection. Serum iron in the control, H. pylori and H. felis groups were significantly (p < .05) elevated at 20 weeks by 39, 26 and 77%, respectively, compared to 4 weeks of infection. H. felis-infected mice had a significantly (p < .05) decreased serum ferritin level during pregnancy (61%) compared to H. pylori infected mice. CONCLUSION: These results suggest that H. felis but not H. pylori infection causes an acute iron deficiency in normal and pregnant mice. PMID- 17083375 TI - Quantitative analysis of representative proteome components and clustering of Helicobacter pylori clinical strains. AB - BACKGROUND: Several Helicobacter pylori proteins have been reported to be associated with severe symptoms of gastric disease. However, expression levels of most of these disease-associated proteins require further evaluation in order to clarify their relationships with gastric disease patterns. Representative proteome components of 71 clinical isolates of H. pylori were analyzed quantitatively to determine whether the protein expression levels were associated with gastric diseases and to cluster clinical isolates. METHODS: After two dimensional electrophoresis (2-DE) of H. pylori isolates, spot intensities were analyzed using pdquest 2-D Gel Analysis Software. The intensities of 10 representative protein spots, identified by peptide fingerprinting using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) or peptide sequencing using quadrupole TOF MS, were subjected to the nonparametric Mann-Whitney test and hierarchical agglomerative cluster analysis. The relationship between clusters and gastric diseases was analyzed by the chi squared test. RESULTS: Although the spot intensities of the 10 representative proteins were highly variable within each gastric disease group, the expression levels of CagA, UreB, GroEL, EF-Tu, EF-P, TagD, and FldA showed some significant differences among the gastric disease patterns. On the basis of the 10 target protein intensities, hierarchical agglomerative cluster analysis generated a dendrogram with clusters indicative of chronic gastritis/gastric cancers and gastric/duodenal ulcers. CONCLUSION: These results indicated that quantitative analysis of proteome components is a feasible method for examining disease associated proteins and clustering clinical strains of H. pylori. PMID- 17083376 TI - Human polymorphonuclear leukocytes are sensitive in vitro to Helicobacter pylori vaca toxin. AB - BACKGROUND: Interactions between bacterial components and polymorphonuclear leukocytes (PMNL) play a major pathogenic role in Helicobacter pylori-associated diseases. Activation of PMNL can be induced by contact with whole bacteria or by different H. pylori products released in the extracellular space either by active secretion or by bacterial autolysis. Among these products, H. pylori VacA is a secreted toxin inducing vacuolation and apoptosis of epithelial cells. METHODS AND RESULTS: We found that non-opsonic human PMNL were sensitive to the vacuolating effect of VacA+ broth culture filtrate (BCF) and of purified VacA toxin. PMNL incubated with VacA+ BCF showed Rab7-positive large intracytoplasmic vacuoles. PMNL preincubation with H. pylori BCF of different phenotypes dramatically potentialized the oxidative burst induced by zymosan, increased phagocytosis of opsonized fluorescent beads, and up-regulated CD11b cell surface expression, but independently of the BCF VacA phenotype. Moreover, by using purified VacA toxin we showed that vacuolation induced in PMNL did not modify the rate of spontaneous PMNL apoptosis measured by caspase 3 activity. CONCLUSIONS: Taken together, these data showed that human PMNL is a sensitive cell population to H. pylori VacA toxin. However, activation of PMNL (i.e., oxidative burst, phagocytosis, CD11b up-regulation) and PMNL apoptosis are not affected by VacA, raising question about the role of VacA toxin on PMNL in vivo. PMID- 17083377 TI - The influence of Helicobacter pylori infection on the prevalence of endoscopic erosive esophagitis. AB - OBJECTIVES: This study aimed to determine the frequency of endoscopic esophagitis and Helicobacter pylori infection in a large Turkish population over a 6-year period. METHODS: We studied a consecutive series of 14,380 patients who had been newly referred for diagnostic esophagogastroduodenoscopy from 2000 to 2006. The mean age value was 45 +/- 10 (18-89) years. All endoscopic findings were retrospectively evaluated. Two antral and two corpus biopsies were taken from patients for rapid urease test. Endoscopic esophagitis was defined as the presence of erosions and/or ulceration. The relationship between erosive esophagitis and various relevant factors was analyzed. RESULTS: The overall prevalence of endoscopic esophagitis was 7.8% (95% CI, 6.9-8.1). The prevalence of positive rapid urease test was 49% (95% CI, 38-53) in patients with esophagitis and 85% (95% CI, 70-96) in patients without esophagitis (p < .001). From 2000 to 2006, the frequency of endoscopic esophagitis and the rate of positive rapid urease test remained unchanged. After adjusting for the effects of mean age, male gender, and percentage of hiatal hernia, there was a 0.785% risk reduction in esophagitis with every 1% increase in the rate of positive rapid urease test result. CONCLUSIONS: The frequency of endoscopic esophagitis is significantly lower in patients with a positive rapid urease test result. This negative correlation with H. pylori infection reflects a protective effect of H. pylori from endoscopic esophagitis in a Turkish population and deserves further investigation. PMID- 17083378 TI - 2-week triple therapy for Helicobacter pylori infection is better than 1-week in clinical practice: a large prospective single-center randomized study. AB - BACKGROUND: Proton pump inhibitor (PPI)-based triple therapies are considered the standard regimens for Helicobacter pylori eradication, but the optimal duration of these regimens is still controversial. The aim of this study was to compare the efficacy of 1-week versus 2-week triple therapies in H. pylori-positive patients. MATERIALS AND METHODS: A total of 486 consecutive H. pylori-positive patients were randomized to receive omeprazole, 20 mg b.i.d., clarithromycin 500 mg b.i.d., and either amoxicillin 1 g b.i.d. or metronidazole 500 mg b.i.d. for 1 or 2 weeks. Upper gastrointestinal endoscopy and histology were performed at entry and 2 months after the end of therapy. H. pylori status was defined according to histology and urea breath test. RESULTS: At intention-to-treat analysis, 2-week therapy with omeprazole, amoxicillin, and clarithromycin achieved a significantly higher eradication rate than 1- or 2-week regimens with metronidazole (70% versus 52%, p = .003, versus 56%, p < .01) and the same therapy for 1-week (70% versus 57%, p = .05). At per-protocol analysis, 2-week therapy with omeprazole, amoxicillin, and clarithromycin showed a significantly higher eradication rate than 1-week of amoxicillin and metronidazole (77% versus 62%; p = .03) but no difference with 1-week same regimen (66%) or 2-week metronidazole and clarithromycin regimen (72%). Compliance and tolerability were good for all regimens. CONCLUSIONS: Two-week therapies, independently of antibiotic combination, lead to a significant increase of H. pylori eradication rate compared to 1-week therapies, with same compliance and tolerability, even if, taking account of low-eradication rates, one must question whether the triple therapy should still be used. PMID- 17083379 TI - A prospective analysis of low-grade gastric malt lymphoma after Helicobacter pylori eradication. AB - BACKGROUND: Primary gastric low-grade B-cell lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) is known to be successfully treated with anti Helicobacter pylori (H. pylori) therapy alone. However, there are few reports on long-term results after eradication therapy. The aims of this study were to analyze the rate and the interval to reach complete remission (CR), and to assess the rate and the factors affecting recurrence of MALT lymphoma. MATERIALS AND METHODS: Between 1996 and 2003, a total of 90 H. pylori-infected patients with low-grade MALT lymphoma were included in this study. For initial staging, endoscopic ultrasonography, chest-abdomen-pelvis CT scans, and bone marrow examination were taken. All patients were made to take anti-H. pylori therapy for 14 days. Tumoral response was assessed by endoscopy every 3 months till CR and every 6 months after achieving CR. RESULTS: Among 90 treated patients, 85 (94.4%) reached CR. The median interval to CR was 3 months (range, 1-24). Seventy-nine (92.9%) patients were in CR at 12 months. Median follow-up period after CR was 45 months (range 15-109). Among 77 patients who were followed-up after CR, 8 (10.4%) patients were proved with recurrence of MALT lymphoma. Cumulative recurrence rate was 2.7, 11.5, and 12.2% at 1, 2, and 3 years. The presence of H. pylori was only a significant risk factor affecting recurrence. CONCLUSIONS: The status of H. pylori is the most important risk factor affecting recurrence. Therefore, adequate eradication regimen and accurate regular evaluation for H. pylori status are needed during follow up of primary gastric low-grade B-cell MALT lymphoma. PMID- 17083380 TI - Prevalence of Helicobacter pylori vacA, cagA, cagE, iceA, babA2 genotypes and correlation with clinical outcome in Turkish patients with dyspepsia. AB - BACKGROUND: Distinct virulence factors of Helicobacter pylori have been associated with clinical outcome of the infection; however, considerable variations have been reported from different geographic regions and data on genotypes of Turkish H. pylori isolates are sparse. AIM: To determine the prevalence of specific genotypes of H. pylori in Turkish patients with dyspepsia. MATERIALS AND METHODS: Ninety-three H. pylori-positive patients [30 with non ulcer dyspepsia (NUD), 30 with duodenal ulcer (DU), and 33 with gastric cancer (GC)] who were admitted to our endoscopy unit due to dyspepsia were enrolled in the study. H. pylori infection was confirmed in all patients by histology and rapid urease test (RUT). The presence of vacA alleles, cagA, cagE, iceA, and babA2 genotypes were determined by polymerase chain reaction (PCR). Chi-squared test and Fisher's exact test were used for statistical comparisons and multivariate regression analysis was performed to find out independent predictors of different clinical outcomes. RESULTS: Turkish strains examined predominantly possessed the vacA s1,m2 (48.4%) and s1,m1 (40.7%) genotypes. The vacA s1a genotype was detected in 66.7, 96.4, and 87.9% of isolates from patients with NUD, DU, and GC, respectively, and its presence was significantly associated with that of DU (p = .004), GC (p = .043), and cagA gene (p = .021). None of the cases was found to harbor the s1c genotype. The frequencies of the cagA and cagE genes among studied isolates were 73.6 and 59.3%, respectively. The cagA gene was significantly associated with the presence of DU (p = .004) and GC (p = .003), and the cagE gene, too, was significantly associated with the presence of DU (p = .002) and GC (p = .000). All H. pylori isolates possessed the iceA gene. In all, 68 isolates (74.7%) were positive for iceA1 and 23 (25.3%) for iceA2. The frequency of icea1 gene was significantly higher in cases with GC (85%) than in cases with NUD (60%) (p = .026). The frequency of babA2 gene was 23.3, 46.4, and 87.9% in isolates of patients with NUD, DU, and GC, respectively. When compared to cases with NUD (p = .000) and DU (p = .000), the presence of babA2 gene was significantly higher in cases with GC. Multivariate regression analysis disclosed cagE (p = .006) and vacA s1a (p = .027) genotypes to be independent predictors of DU and babA2 (p = .000) and cagE (p = .013) genotypes to be independent predictors of GC. CONCLUSIONS: H. pylori vacA s1a, cagA, cagE genotypes have significant relations with the presence of DU and GC, and iceA1, babA2 with GC in Turkish patients with dyspepsia, whereas cagE and vacA s1a genotypes are independent predictors of DU, and babA2 and cagE genotypes are independent predictors of GC. PMID- 17083381 TI - Alaska sentinel surveillance for antimicrobial resistance in Helicobacter pylori isolates from Alaska native persons, 1999-2003. AB - BACKGROUND: Previous studies in Alaska have demonstrated elevated proportions of antimicrobial resistance among Helicobacter pylori isolates. MATERIALS AND METHODS: We analyzed H. pylori data from the Centers for Disease Control and Prevention (CDC)'s sentinel surveillance in Alaska from July 1999 to June 2003 to determine the proportion of culture-positive biopsies from Alaska Native persons undergoing routine upper-endoscopy, and the susceptibility of H. pylori isolates to metronidazole [minimum inhibitory concentration (MIC) of > 8 g metronidazole/mL), clarithromycin (MIC > or = 1), tetracycline (MIC > or = 2) and amoxicillin (MIC > or = 1)] using agar dilution. RESULTS: Nine-hundred sixty-four biopsy specimens were obtained from 687 participants; 352 (51%) patients tested culture positive. Mean age of both culture-positive and culture-negative patients was 51 years. Metronidazole resistance was demonstrated in isolates from 155 (44%) persons, clarithromycin resistance from 108 (31%) persons, amoxicillin resistance from 8 (2%) persons, and 0 for tetracycline resistance. Metronidazole and clarithromycin resistance varied by geographic region. Female patients were more likely than male subjects to show metronidazole resistance (p < .01) and clarithromycin resistance (p = .05). CONCLUSIONS: Resistance to metronidazole and clarithromycin is more common among H. pylori isolates from Alaska Native persons when compared with those from elsewhere in the USA. PMID- 17083382 TI - Effect of the folk remedy, Bainiku-ekisu, a concentrate of Prunus mume juice, on Helicobacter pylori infection in humans. AB - BACKGROUND: Bainiku-ekisu, a concentrate of Japanese apricot (Prunus mume) juice, is a traditional Japanese folk remedy for treatment of dyspepsia since more than a thousand years ago. Fujita et al. previously reported in vitro antibacterial effect of Bainiku-ekisu to Helicobacter pylori. We conducted an in vivo pilot study to evaluate the possibility that Bainiku-ekisu may have an antibacterial effect on H. pylori in the human stomach. MATERIALS AND METHODS: Consecutive 18 H. pylori-positive subjects were included. Approximately 130 mL 1% Bainiku-ekisu solution was ingested by the subjects twice a day for 12 weeks. Urea breath test (UBT) was performed before ingestion, and 2 and 12 weeks after starting ingestion of Bainiku-ekisu, and UBT values were compared. RESULTS: Bainiku-ekisu therapy resulted in a slight fall in UBT values after 2 weeks (from 30.1 +/- 6 to 23.5 +/ 6 in ITT analysis, p = .094; from 31.2 +/- 6 to 24.7 +/- 6 in PP analysis, p = .124) (data are shown with mean +/- SE). In two instances (11%), the UBT values became negative. Fourteen subjects completed the trial for 12 weeks and there was no significant change in UBT values (from 30.1 +/- 6 to 25.9 +/- 6 in ITT analysis, p = .450; from 35.6 +/- 6 to 31.4 +/- 7 in PP analysis, p = .555). CONCLUSION: Our results are consistent with the antibacterial effect of Bainiku ekisu on H. pylori in the human stomach. However, the bacteria were not successfully eradicated with 2- or 12-week ingestion of a Bainiku-ekisu solution. Subsequent studies will need to identify a clinically useful regimen. PMID- 17083384 TI - Impact of thrombocytosis and C-reactive protein elevation on the prognosis for patients with renal cell carcinoma. AB - AIM: C-reactive protein (CRP) elevation is reportedly a prognostic factor in patients with renal cell carcinoma (RCC). Thrombocytosis has recently been reported also to be a prognostic factor in RCC and, like CRP, to be related to inflammatory cytokines such as interleukin-6. The aim of this study was to evaluate the importance of both thrombocytosis and CRP elevation in tumor recurrence and prognosis for patients with RCC. METHODS: The clinical records of 178 patients who underwent radical nephrectomy were reviewed. Thrombocytosis was defined as a platelet count >or=350,000/mm(3), and CRP elevation was defined as a CRP level >or=1.0 mg/dL. Disease-free survival and cause-specific survival rates were calculated. Independent predictors for recurrence and prognosis were determined. RESULTS: Patients with thrombocytosis and patients with elevated CRP levels had significantly higher pathological T stage, clinical stage, tumor size, histological grade, and percentage of microvascular invasion than did patients without THC and patients with CRP levels <1.0 mg/dL, respectively. There was a significant correlation between platelet counts and CRP levels. Multivariate analysis showed that distant metastasis, tumor size, grade 3 components, and CRP elevation were independent predictors for prognosis but thrombocytosis was not. In N0M0 RCC patients, tumor size, microvascular invasion, and CRP elevation were independent predictors for recurrence. CRP elevation and tumor size were independent predictors for prognosis. CONCLUSIONS: Platelet count and CRP level are strongly correlated in patients with RCC, but only CRP elevation is an independent predictor for recurrence and prognosis. PMID- 17083385 TI - Evaluation of renal function after laparoscopic partial nephrectomy with renal scintigraphy using 99mtechnetium-mercaptoacetyltriglycine. AB - AIM: We evaluated the functions of an affected kidney after laparoscopic partial nephrectomy (LPN) using renal scintigraphy with (99m)technetium mercaptoacetyltriglycine ((99m)Tc-MAG3). METHODS: Split renal function of 10 patients who underwent LPN for renal tumors was assessed using renal scintigraphy with (99m)Tc-MAG3 before surgery, and 1 week and 3 months post-surgery. RESULTS: Median operating time was 196.5 min, median tumor diameter was 2.3 cm, mean blood loss was 64 mL and mean ischemic time was 38.5 min. Median change in serum creatinine level pre- to post-surgery was 0.15 mg/dL. Median contribution of the affected kidney to total renal function (calculated using (99m)Tc-MAG3) was 50.0%, 41.7% and 36.1% before surgery, 1 week and 3 months after LPN, respectively. In one patient, the tumor was resected after cooling of the affected kidney with ice slush for 15 min, and the split renal function ratio remained as high as 50% at 3 months post-operatively despite a total ischemic time of 61 min. CONCLUSIONS: This paper evaluated renal function on the affected side before and after surgery by measuring split renal function with renal scintigraphy using (99m)Tc-MAG3. Risk factors for renal dysfunction in the affected kidney after LPN include age over 70 years with more than 30 min warm ischemic time, re-clamping of the renal artery procedure, and a warm ischemic time greater than 60 min. We believe that renal cooling with slush ice prevents renal dysfunction of the affected kidney after LPN with longer warm ischemic times. However, an easier renal cooling technique should be sought for regular use of cooling procedures in LPN. PMID- 17083386 TI - Percutaneous ultrasound-guided radiofrequency ablation for kidney tumors in patients with surgical risk. AB - OBJECTIVE: The aim of this study was to describe our experience with percutaneous ultrasound-guided radiofrequency ablation of kidney tumors. METHODS: From July 2002 to August 2005, 45 radiofrequency ablations (RFA) in 42 selected patients with kidney tumor were performed. The patients had either contraindications to surgery procedures or had a solitary kidney. The average tumor size was 37.5 mm (range, 18-59 mm) with the mean age of 68 years (range, 28-83 years). RFA were performed based on radiographic findings. Needle biopsy was made only twice. Monopolar Cool-tip Tyco or bipolar Celon Olympus radiofrequency devices were used. The procedure was performed under conscious sedation with local anesthesia. Treatment efficacy was assessed by computed tomography and by Doppler ultrasound. The absence of contrast enhancement on computed tomography was considered to be a successful treatment. RESULTS: The average follow up was 14 months (range, 3-36 months). In 42 tumors (93%), total absence of contrast enhancement was obtained after the initial RFA and in three tumors (7%) after the second ablation session. There were no complications following 41 procedures, including all ablations in small (<35 mm) renal masses. In four procedures, minor complications were observed. All patients are alive. There has been no need for chronic hemodialysis and, until now, we have not observed any local recurrences with the exception of one metastasis to an ipsilateral adrenal gland. CONCLUSIONS: RFA of kidney tumors is a promising alternative treatment which could be considered for patients who are not suitable for surgery. PMID- 17083387 TI - Elevation of serum and urinary carbohydrate antigen 19-9 in benign hydronephrosis. AB - OBJECTIVE: The aim of this study was to find out the predictive role of serum and urinary carbohydrate antigen 19-9 levels in benign hydronephrosis and whether these levels are helpful for differentiation of complete or partial urinary obstruction. MATERIALS AND METHODS: Fifty-four patients with, and 23 without, benign hydronephrosis were enrolled in this study. Serum and urinary carbohydrate antigen 19-9 were determined by the chemiluminescence enzyme immunometric assay method and these levels were correlated with clinical factors. RESULTS: The mean serum (P < 0.0001) and urinary (P < 0.0001) carbohydrate antigen 19-9 and serum creatinine (P < 0.008) levels were significantly higher in the hydronephrosis group than the control group. There was significant correlation between serum and urinary carbohydrate antigen 19-9 levels in the hydronephrosis group (r = 0.639, P < 0.0001). In the hydronephrosis group, there were no significant differences between the serum creatinine, serum or urinary carbohydrate antigen 19-9 levels and the clinical features except symptom duration. The best cut-off value for the serum and urinary carbohydrate antigen 19-9 were found to be 4.84 U/mL and 29.35 U/mL, respectively. CONCLUSION: Serum and urinary carbohydrate antigen 19-9 levels correlated with each other, were significantly elevated in patients with hydronephrosis and did not predict complete urinary obstruction. Benign hydronephrosis should be considered in the differential diagnosis of carbohydrate antigen 19-9 increments, as this is what is of most importance in clinical practice. PMID- 17083388 TI - When is open ureterolithotomy indicated for the treatment of ureteral stones? AB - AIM: Improvements in extracorporeal shock wave lithotripsy (ESWL) and ureteroscopy have almost eradicated the need for open surgery in ureteral stones. The aim of this study was to assess characteristics of patients who underwent open ureterolithotomy. METHODS: During a 5-year period, a total of 654 patients with ureteral stones were treated. Initial management consisted of ureteroscopy in 524 patients, ESWL in 62 patients and percutaneous nephrolithotomy (PCNL) in 12 patients. Open surgery was performed in 56 patients. Stone location, size and success rates were retrospectively analyzed. RESULTS: Ureteroscopy resulted in successful stone removal in 94%, 98% and 98.5% of proximal, mid and distal ureteral stones, respectively. A total of 14 patients with ureteroscopy failure were referred for open surgery. ESWL treatment resulted in success in 55 patients (88%), and those with ESWL failure were referred for either ureteroscopy (n = 3) or open surgery (n = 4). Open surgery was performed in a total of 56 patients, 38 of whom had been referred from other centers. Stone location was proximal ureter in 25 (44.6%) patients (stone size: 2-12 cm(2)), mid ureter in five (8.9%) patients (stone size: 2-6 cm(2)) and distal ureter in 26 (46.4%) patients (stone size: 4-9 cm(2)). A history of previous unsuccessful endourological procedure was observed in 33 (58%) of 56 patients. Children under age 16 (range 1-15 years) comprised 17.8% of patients undergoing open surgery. CONCLUSION: Open surgery, which is nowadays being replaced with laparoscopic techniques, is generally indicated for failed endourological procedures (58%), particularly in centers that do not have flexible ureteroscopy or laser lithotriptor, and in patients with larger stones (>3 cm). Children (17.8%) are also candidates for open surgery, if specifically designed endourological equipment is not available. PMID- 17083389 TI - Analysis of factors predicting intravesical recurrence of superficial transitional cell carcinoma of the bladder without concomitant carcinoma in situ. AB - BACKGROUND: The objective of this study was to investigate risk factors for intravesical recurrence in patients with superficial bladder cancer without concomitant carcinoma in situ (CIS). METHODS: In this series, we analyzed data from patients with newly diagnosed superficial Ta or T1 transitional cell carcinoma (TCC) of the bladder without concomitant CIS who underwent complete transurethral resection (TUR) without any adjuvant intravesical instillation therapies. Multivariate analysis was used to determine significant risk factors affecting intravesical recurrence after TUR. Differences in clinicopathological features between primary and recurrent tumors were also characterized. RESULTS: Among 341 patients undergoing TUR of Ta or T1 bladder cancer, 187 diagnosed as having concomitant CIS and/or treated with adjuvant intravesical therapy were excluded, and the remaining 154 were evaluated. Intravesical recurrence was detected in 64 of the 154 patients, showing a 5-year recurrence-free survival rate of 58.3%. Among several factors examined, only tumor size was significantly associated with intravesical recurrence. Multivariate analysis identified tumor size as an independent predictor for intravesical recurrence irrespective of other parameters including age, gender, multiplicity, growth pattern, grade and stage. Recurrent tumors were significantly smaller and of a lower grade and lower stage than primary tumors, despite the absence of differences in growth pattern and the multiplicity between them. CONCLUSIONS: These findings suggest that primary tumor size could be used as a potential risk factor for predicting intravesical recurrence following TUR of superficial TCC of the bladder without concomitant CIS, and that the pathological characteristics of recurrent tumors are more favorable than those of primary tumors. PMID- 17083390 TI - Outcome of conservative treatment of primary vesicoureteral reflux in 87 Thai children in a single center. AB - AIM: To evaluate the outcome of primary vesicoureteral reflux (VUR) using conservative treatment. METHODS: Eighty-seven children with primary VUR who had been treated with a conservative medical regimen and monitored through a yearly cystogram were recruited for the study. The study was conducted at the Pediatric Nephrology Clinic in Songklanagarind Hospital, the major tertiary care center in southern Thailand. Statistical analyses using Kaplan-Meier survival curves, chi squared test, Fisher's exact test and multivariate analysis with Cox regression were performed. RESULTS: The study group consisted of 41 boys and 46 girls, with a total of 133 VUR. The age of the boys was significantly lower than that of the girls (P < 0.001). Resolution of the low grades (grades I-III) of VUR was significantly more frequent than that of the high grades (grades IV-V) (68/95, 72%vs 14/38, 37%; P < 0.001). Using the Kaplan-Meier survival analysis and log rank test, the resolutions of VUR in boys versus girls, and age <1 years versus >or=1 years were not significantly different (P = 0.2252 and 0.4756, respectively). Low-grade VUR and unilateral VUR had significantly higher probabilities of resolution than high-grade VUR and bilateral VUR, respectively (P = 0.0041 and 0.0467, respectively). Multivariate Cox regression analysis indicated that among low-grade VUR, boys and unilateral VUR had significantly higher probabilities of resolution. Among the high-grade VUR, neither sex nor laterality had a significant effect on the probabilities of VUR resolution. CONCLUSION: This study offers more evidence that children with low-grade VUR have a higher chance of reflux resolution if they are boys and have only unilateral VUR. There is still no good guide for resolution of reflux in children with high grade VUR. PMID- 17083391 TI - Is the short-term outcome of transurethral resection of the prostate affected by preoperative degree of bladder outlet obstruction, status of detrusor contractility or detrusor overactivity? AB - AIM: The aim of this study was to investigate whether the preoperative degree of bladder outlet obstruction (BOO), detrusor underactivity (DUA) or detrusor overactivity (DO) affected the short-term outcome of transurethral resection of the prostate (TURP) for patients with lower urinary tract symptoms suggestive of benign prostatic hyperplasia (LUTS/BPH). METHODS: Ninety-two patients with LUTS/BPH aged 50 years or older who were considered to be appropriate candidates for TURP were included in this study. Pressure-flow study and filling cystometry were performed to determine BOO, DUA and DO before TURP. The efficacy of TURP was determined at 3 months after surgery using the efficacy criteria for treatment of BPH assessed by the International Prostate Symptom Score, QOL index, maximum flow rate and postvoid residual urine volume. RESULTS: On preoperative urodynamics, 60%, 40% and 48% of patients showed BOO, DUA and DO, respectively. After TURP, 76% showed 'excellent' or 'good' overall efficacy, whereas only 13% fell into the 'poor/worse' category. The efficacy was higher as the preoperative degree of BOO worsened. In contrast, neither DO nor DUA influenced the outcome of TURP. However, the surgery likely provided unfavorable efficacy for patients having DO but not BOO. Only 20% of the patients who had both DO and DUA but did not have BOO achieved efficacy. CONCLUSIONS: Transurethral resection of the prostate is an effective surgical procedure for treatment of LUTS/BPH, especially for patients with BOO. DUA may not be a contraindication for TURP. The surgical indication should be circumspect for patients who do not have BOO but have DO. PMID- 17083392 TI - Efficacy and safety of tamsulosin hydrochloride compared to doxazosin in the treatment of Indonesian patients with lower urinary tract symptoms due to benign prostatic hyperplasia. AB - AIM: The objective of the study was to compare the efficacy and safety of tamsulosin hydrochloride and doxazosin in patients with lower urinary tract symptoms (LUTS) due to benign prostatic hyperplasia (BPH). METHODS: The safety and efficacy of tamsulosin (0.2 mg) and doxazosin (2 mg) was determined after once daily administration for 6 weeks in an open-label, randomized, multicenter study of 101 men with BPH. The International Prostatic Symptom Score (IPSS), maximal urinary flow rates (Qmax), average urinary flow rates (Qave) and residual urine were determined at baseline and again at 6 weeks as efficacy parameters. The primary parameters used for safety evaluation were vital signs (blood pressure and heart rate) and adverse events. The number of patients with a clinically significant response to treatment with tamsulosin or doxazosin was determined and defined as those with >20% improvement from the baseline Qmax or >20% decrease in total IPSS. RESULTS: The total IPSS decreased significantly in both the tamsulosin and doxazosin groups compared to baseline. There was a significant difference in the decrease in total IPSS between two groups. Qmax, Qave and residual urine significantly improved only in the tamsulosin group. There were no significant differences in systolic blood pressure, diastolic blood pressure or heart rate profile in the tamsulosin group; however, doxazosin resulted in a significant difference in systolic and diastolic blood pressure. Tamsulosin was well tolerated; only three patients (6%) in the tamsulosin group reported an adverse event (dizziness) while 11 patients (22%) in the doxazosin group reported an adverse event (dizziness), one of whom withdrew from the study. CONCLUSIONS: Tamsulosin was shown to be more effective than doxazosin in the treatment of LUTS due to BPH. PMID- 17083393 TI - One year follow-up on the SPARC sling system for the treatment of female urodynamic stress incontinence. AB - AIM: To evaluate the safety and efficacy of the SPARC sling in women with urinary incontinence. METHODS: Women who presented with urinary incontinence symptoms were included in this prospective study. In total, 103 women underwent evaluation of subjective and objective cure rates, quality of life and urodynamic parameters following the SPARC procedure. RESULTS: The objective cure rate, defined as a pad weight test less than 1 g and negative cough stress test, was 84.4% at 12 months. Subjective cure rates, defined as no usage of pads/no urine loss during daily activities, were also encouraging: 75.0% at 12 months. Quality of life evaluation included asking patients to rate the amount of bother caused by their incontinence symptoms on a scale from 0 to 100. Preoperatively, the average was 76.7, dropping to 15.9 at 12 months. At 6 months, 93.7% (75/79) were 'very satisfied' and 5.1% (4/79) were 'satisfied;' one patient was 'dissatisfied' (1.2%, 1/79). Seventy-eight of 79 patients (98.7%) would recommend the procedure to a friend and undergo the procedure again. Patients returned to inside work an average of 1.4 days and to outside work 5.4 days after the procedure. The maximum flow rate decreased from a mean of 43.4 mL/s preoperatively to 37.08 mL/s at 12 months postoperatively (P = 0.049). Detrusor pressure at maximum flow and maximal urethral closure at rest increased from 16.94 and 50.85 cmH(2)O preoperatively to 21.69 and 60.61 cmH(2)O at 12 months, respectively (P = 0.015; P < 0.001). There were few postoperative complications, with no reported infections, erosions or hematomas. CONCLUSIONS: This prospective study demonstrates that the SPARC suprapubic sling is a safe and effective treatment for female stress urinary incontinence. Both objective and subjective measures of success were achieved, with low complication rates. Quality of life and urodynamic parameters were improved. The SPARC method provides safe and highly effective treatment for female stress urinary incontinence at one year. PMID- 17083394 TI - Dorsal onlay augmentation urethroplasty with small intestinal submucosa: modified Barbagli technique for strictures of the bulbar urethra. AB - AIM: To present the results from one clinic's experience of using small intestinal submucosa (SIS) in augmentation urethroplasty for management of strictures of the bulbar urethra. METHODS: Urethral surgery was performed in nine men with strictures 4-6 cm. All of the patients were evaluated by history, physical examination, retrograde urethrogram, and uroflowmetry. Four layers of SIS were soaked in saline or Ringer's solution for 15 minutes at 37 degrees C, and the inner surface of the patch was gently fenestrated with a thin scalpel. The patch was spread-fixed onto the tunica albuginea. The mucosa was sutured to the submucosal graft first at 2-3 mm inwards from the SIS margins, then the spongiosum tissue was attached to the margins with interrupted absorbable sutures. RESULTS: Of the nine patients who underwent augmentation urethroplasty using SIS, only one had re-stricture at 6 months due to urethral infection. At 18 months after the surgery the uroflowmetry of the other eight patients was 20-21 mL/s. In terms of complications, six patients reported having post-micturition dribbling, and seven patients reported lack of morning erections for 35-69 days after surgery. CONCLUSIONS: Using SIS is a safe procedure; however, long-term follow-up is needed to substantiate the good short-term results. PMID- 17083395 TI - Failure to detect urethral Trichomonas vaginalis in Japanese men with or without urethritis. AB - AIM: Trichomonas vaginalis may cause symptomatic or asymptomatic urethritis in men. There are few recent studies on the prevalence of T. vaginalis infection in Japanese men, and quantification of the number of cases of urethritis attributable to this pathogen has not been performed in Japan. The aim of this study was to determine the prevalence and morbidity of T. vaginalis infection in Japanese men. METHODS: One hundred subjects with or without urethritis were examined for the presence of urethral T. vaginalis using culture swabs. RESULTS: Urethral swabs from all subjects were negative for T. vaginalis. CONCLUSION: These results indicate Japanese men, including those with urethritis, have a low incidence of urethral T. vaginalis infection or colonization. T. vaginalis appears to be an uncommon pathogen for male urethritis in Japan. PMID- 17083396 TI - Laparoscopic versus open orchiopexy in the management of abdominal testis: a descriptive study. AB - AIMS: Treatment of patients with abdominal non-palpable testis (NPT) is still controversial among pediatric urologists. This is a prospective randomized comparative study between open and laparoscopic orchiopexy for management of abdominal testis. The aim of this study was to evaluate the success rate and morbidity of both approaches. METHODS: Eighty-two patients with a mean age of 5.3 years were evaluated by laparoscopy for 87 NPT. Patients with viable abdominal testes were randomly treated with either open or laparoscopic orchiopexy procedures. RESULTS: On laparoscopy, 75 viable abdominal testes were found. According to location: 41 (47.1%) testes were high abdominal, 27 (31%) testes were low abdominal and 7 (8%) testes were peeping from the internal ring. Laparoscopic first stage Fowler-Stephens orchiopexy was done initially for those with high abdominal testes. For further management, all patients were divided randomly into open (36 cases) and laparoscopic (39 cases) groups where primary (with spermatic vessel preservation) or second stage Fowler-Stephens orchiopexy was done. Statistical analysis was done using Student's t-test. Laparoscopic procedures showed significant less morbidity than the open counterparts. Follow up ranged from 9 to 31 months and included evaluation of testicular site and size. All testes were located satisfactorily inside the scrotum. Five cases of testicular atrophy were encountered (three and two testes with open and laparoscopic second stage Fowler-Stephens orchiopexy respectively) after 1 year follow up. CONCLUSION: Results of open versus laparoscopic orchiopexy procedures (primary or staged) are fairly comparable. However, laparoscopy provides significantly less morbidity. PMID- 17083397 TI - New onset of hydroceles in boys over 1 year of age. AB - AIM: The presentation, and medical and surgical management of all new onset non congenital hydroceles in boys older than 1 year of age were examined. Of particular interest was the outcome of those patients who presented with a non communicating hydrocele that developed after the first year of life and was managed conservatively. METHODS: All patients older than 12 months of age who were evaluated as outpatients with the diagnosis of hydrocele from January 1994 to January 2001 were identified. Possible risk factors and predisposing conditions were determined. For the patients who had surgical correction, surgical indications were identified. For non-surgical patients, long-term outcomes were recorded. RESULTS: A total of 302 patients older than 12 months of age with the diagnosis of new onset hydrocele were identified. Of these, 35% were non-communicating, 59% were communicating, and 6% were hydroceles of the spermatic cord. In terms of surgery, 97% of communicating hydroceles, 71% of hydroceles of the spermatic cord, and 34% of non-communicating hydroceles had operative management. Seventy patients with non-communicating hydroceles did not receive surgery and 51 (73%) were contacted for long term follow-up. In these 51 patients, 76% of non-communicating hydroceles resolved completely, 6% decreased in size but were still present, 14% remained the same size, and 4% had an unknown status. The average time to resolution was 5.6 months with a median time of 3 months. The time range to resolution was from 1 day to 24 months. Follow-up averaged 73.7 months with a range of 33 to 120 months. CONCLUSIONS: Approximately 75% of new onset, non-congenital, non-communicating hydroceles resolve spontaneously irrespective of size. An observation period of 6-12 months would be appropriate prior to repair. PMID- 17083398 TI - Up-titration of vardena fi l dose from 10 mg to 20 mg improved erectile function in men with spinal cord injury. AB - AIM: Vardenafil is a highly selective phosphodiesterase type-5 inhibitor for the treatment of erectile dysfunction (ED). Efficacy of vardenafil has been demonstrated in various ED populations, but that in Japanese patients with spinal cord injury (SCI) has not been assessed. METHODS: This was an open-label, multicenter, flexible dose, 12-week study in patients with ED due to SCI. Following a 4-week observation period, patients received vardenafil 10 mg for 4 weeks, and based on efficacy, tolerability and patient preference, doses for the remaining 8 weeks were decided by investigators. The primary efficacy parameter was erectile function domain score of the International Index of Erectile Function. RESULTS: Ten patients took 10 mg all through the study, while 22 patients took 20 mg after completing 4 weeks' treatment with 10 mg. The erectile function domain score increased from 12.2 at baseline to 25.0 at Last Observation Carried Forward (LOCF) in the former group and from 10.3 to 22.5 in the latter group, respectively. Importantly, there was a 5.0 point increase in erectile function domain score after up-titration in the latter group. Drug-related adverse events were observed in 22% of patients including hot flushes (9%) and headache (6%), but these were transient and mild in intensity. Serious adverse events and adverse events leading to discontinuation of the study drug were not reported. CONCLUSIONS: Vardenafil 10 and 20 mg was well tolerated and improved erectile function in patients with SCI. Of interest, erectile function was further improved by 20 mg in patients who were not sufficiently treated with 10 mg. PMID- 17083399 TI - Prevention of renal ischemia/reperfusion-induced injury in rats by leflunomide. AB - OBJECTIVE: There is increasing evidence to suggest that toxic oxygen radicals play an essential role in the pathogenesis of ischemia/reperfusion (I/R) injury in the kidney. This study was designed to investigate the effects of leflunomide, an isoxazole derivative and a unique immunomodulatory agent, in I/R-induced renal injury in rats. METHODS: Forty female Sprague-Dawley rats were divided equally into four groups: (I) control (only leflunomide 10 mg/kg, intragastrically treated); (II) sham operated (only unilateral nephrectomy); (III) I/R; and (IV) leflunomide (10 mg/kg for two doses prior to experiment) plus I/R groups. In groups III and IV, after unilateral nephrectomy, the rats were subjected to 60 min of left renal pedicle occlusion, followed by 6 h of reperfusion. At the end of the reperfusion period, rats were killed and kidneys and blood were removed. Catalase, myeloperoxidase and xanthine oxidase activities, and malondialdehyde, nitric oxide and protein carbonyl levels were determined in renal tissue. Serum creatinine, blood urea nitrogen and aspartate aminotransferase were measured for the evaluation of renal function. In histopathological examination, renal damage was scored 0-3. RESULTS: Group III animals demonstrated severe deterioration of renal function, renal morphology and a significant renal oxidative stress. Pretreatment of animals with leflunomide markedly attenuated renal dysfunction, morphological alterations, reduced elevated oxidative stress products levels and restored the depleted renal antioxidant enzyme. CONCLUSION: The findings imply that oxygen radicals play a causal role in I/R-induced renal injury, and leflunomide exerts renoprotective effects probably by the radical scavenging and antioxidant activities with immunomodulatory effect. PMID- 17083400 TI - Idiopathic retroperitoneal fibrosis associated with IgG4-positive-plasmacyte infiltrations and idiopathic chronic pancreatitis. AB - Idiopathic retroperitoneal fibrosis (IRPF) is an inflammatory fibrosclerosing condition, leading to renal failure by obstruction of the ureters. Idiopathic chronic pancreatitis associated with marked inflammatory infiltrates has recently been referred to as autoimmune pancreatitis (AIP), and infiltrating plasmacytes carrying immunoglobulin-gamma type 4 (IgG4) are relevant to its pathogenesis. The case is described herein of IRPF associated with subclinical pancreatitis that was most probably AIP in a 70-year-old man. Biopsy specimens of the retroperitoneal pseudotumor revealed a marked lymphoplasmacytic infiltration with dense fibrosis. Infiltrating plasma cells were immunoreactive for anti-IgG4 antibodies. Subsequent systemic examinations showed an extremely elevated serum IgG4 level and pancreatitis concordant with AIP. Following oral steroid administration, the serum IgG4 level normalized, although the appearance of the pseudotumor did not alter. Some AIP cases have been associated with idiopathic fibrosclerosing disorders including IRPF, but histological evidence of IgG4 related IRPF has rarely been provided. PMID- 17083401 TI - Development of multiple calculi in the duplex system ureterocele. AB - A 61-year-old man presented complaining of pollakisuria and nocturia. A plain radiograph of his kidney, ureter and bladder and intravenous urography revealed numerous calculi in the upper kidney of his left renal pelvis and ureterocele. A transurethral incision of ureterocele (TUI-ureterocele) and extracorporeal shock wave lithotripsy were performed. On TUI-ureterocele, the many calculi were found to be almost the same size and spherical in form. The postoperative clinical course was uneventful. PMID- 17083402 TI - Urothelial carcinoma (clear cell variant) diagnosed with useful immunohistochemistry stain. AB - The case is reported of urothelial carcinoma (clear cell variant) that was diagnosed with useful immunohistochemistry stain. A 70-year-old man, who had undergone left radical nephrectomy for renal cell carcinoma in August 2003 and partial lobectomy for pulmonary metastasis in May 2005, complained of hematuria in June 2005. On evaluation, a papillary pedunculated tumor was detected in the left wall of the urinary bladder. A transurethral resection of the bladder tumor (TUR-Bt) was performed in July 2005. The pathological diagnosis was difficult due to diffuse clear cell appearance. Immunohistochemistry stain showed urothelial carcinoma, not metastasis of the renal cell carcinoma. Finally it was diagnosed as urothelial carcinoma clear cell variant. Urothelial carcinoma has many variants that show a variety of appearances and characteristics. These should be well known before medical therapy is initiated. PMID- 17083403 TI - Gastrointestinal stromal tumor in ileal neobladder. AB - Gastrointestinal stromal tumor (GIST) is a recently described mesenchymal tumor that can develop in any portion of the gastrointestinal tract. The occurrence of a GIST in the urinary tract is rare, but GIST can present as tumor of the urinary tract or invade the urinary tract. This is the first reported case of GIST in the ileal neobladder, which presented as a submucosal tumor. The patient underwent an open exploration and partial resection of the neobladder pouch. PMID- 17083404 TI - Micturitional disturbance due to labial adhesion as a cause of vaginal implantation of bladder urothelial carcinoma. AB - Vaginal implantation of urinary tract urothelial carcinoma is a rare finding, with few cases reported in the literature. This is the first reported case of vaginal implantation of bladder urothelial carcinoma thought to be due to micturitional disturbances secondary to labial adhesion. The authors propose that implantation via pooled urine in the vagina may have occurred, and suggest that labial adhesion be treated in patients with urinary tract urothelial carcinoma, even if asymptomatic. PMID- 17083405 TI - Radiofrequency ablation partial nephrectomy: a new method of nephron-sparing surgery in selected patients. AB - From July 2002 to April 2005, seven radiofrequency ablation partial nephrectomies have been carried out in seven selected patients. A cool-tip Tyco radiofrequency device under intraoperative ultrasound guidance was used. After intervention, tumors were removed and their tissue with their margins were verified histopathologically. Procedure efficacy was assessed by multidetector computed tomography and by ultrasound. Complications included urine leakage in three cases. Histopathologically, in every case renal cell carcinoma was detected. There is no need for dialysis and there has been no tumor recurrence. No bleeding without clamping renal pedicle, easy tumor extraction and, we hope, reduced risk of recurrence are the major advantages of this intervention. PMID- 17083406 TI - Nurses in the forefront of tuberculosis prevention, care and treatment. PMID- 17083407 TI - Tuberculosis: nurses' work continues. PMID- 17083411 TI - Social advocacy and helping others are fundamental to nursing. PMID- 17083412 TI - Reflections on celebrating AIDS day with fanfare. PMID- 17083413 TI - Nurses as providers of emotional support to patients with MDR-TB. AB - AIM: To identify the forms and means of emotional support that nurses provide to patients living with multidrug-resistant tuberculosis (MTR-TB) in Lima, Peru. BACKGROUND: A fundamental role of nurses is to provide emotional support, defined as all the strategies that a health team employs to assure the psychosocial well being of the patient. However, neither the forms of emotional support nor the means used by nurses in resource-poor settings have been much written about. This paper describes a qualitative study of a team of seven nurses working in a programme that provides individualized MDR-TB treatment to patients in Lima, Peru. It describes the various forms of support that facilitated the ability of patients to adhere to treatment despite socio-economic difficulties, social stigma, drug side effects, problems related to different stages of treatment and concurrent illnesses/special situations. METHODS: Qualitative study methods were employed over the course of 8 years to observe nurses and patients in an MDR-TB treatment programme. These included participant observation, structured observation sessions of nurses with their patients and focus groups with seven nurses. CONCLUSION: Through theme and content analyses of qualitative data, ten situations related to MDR-TB treatment were found. These ten issues served as an analytical framework used to identify and discuss the various types of emotional support provided by both formal and informal means. This type of support focused on problems related to different stages of treatment, social stigma of the illness, treatment adherence, side effects, socio-economic difficulties, death and concurrent illnesses/special situations. PRACTICE IMPLICATIONS: The essential role of the nurse as a provider of emotional support in the development or implementation of similar programmes with MDR-TB should, in future, be taken into account. PMID- 17083414 TI - The Practical Approach to Lung Health in South Africa (PALSA) intervention: respiratory guideline implementation for nurse trainers. AB - AIM: This paper describes the design, facilitation and preliminary assessment of a 1-week cascade training programme for nurse trainers in preparation for implementation of the Practical Approach to Lung Health in South Africa (PALSA) intervention, tested within the context of a pragmatic cluster randomized controlled trial in the Free State province. PALSA combines evidence-based syndromic guidelines on the management of respiratory disease in adults with group educational outreach to nurse practitioners. BACKGROUND: Evidence-based strategies to facilitate the implementation of primary care guidelines in low- to middle-income countries are limited. In South Africa, where the burden of respiratory diseases is high and growing, documentation and evaluation of training programmes in chronic conditions for health professionals is limited. METHOD: The PALSA training design aimed for coherence between the content of the guidelines and the facilitation process that underpins adult learning. Content facilitation involved the use of key management principles (key messages) highlighted in nurse-centred guidelines manual and supplemented by illustrated material and reminders. Process facilitation entailed reflective and experiential learning, role-playing and non-judgemental feedback. DISCUSSION AND RESULTS: Preliminary feedback showed an increase in trainers' self-awareness and self confidence. Process and content facilitators agreed that the integrated training approach was balanced. All participants found that the training was motivational, minimally prescriptive, highly nurse-centred and offered personal growth. CONCLUSION: In addition to tailored guideline recommendations, training programmes should consider individual learning styles and adult learning processes. PMID- 17083415 TI - Awareness and determinants of family planning practice in Jimma, Ethiopia. AB - BACKGROUND: The continuing growth of the world population has become an urgent global problem. Ethiopia, like most countries in sub-Saharan Africa, is experiencing rapid population growth. Currently, the country's population is growing at a rate of 3%, one of the highest rates in the world and if it continues unabated, the population will have doubled in 23 years, preventing any gain in the national development effort. AIM: To determine the level and determinants of family planning awareness and practice in one Ethiopian town. METHODOLOGY: A quantitative study using a descriptive survey design was conducted in Jimma University Hospital. DISCUSSION: The findings revealed that the knowledge and practice of modern contraception methods was low. Most women's contraceptive knowledge and practice was influenced by socio-cultural norms such as male/husband dominance and opposition to contraception, and low social status of women. A lack of formal education for women was identified as a key factor in preventing change in the patterns of contraceptive knowledge and use by women in this part of Ethiopia. CONCLUSION: The support and encouragement for women and men to enter and complete formal education is essential in bringing about a cultural and social change in attitude towards the economic and social value of family planning. This study and others suggest that education can address the imbalance in decision making about contraception and the role of women in society generally. PMID- 17083416 TI - The international migration of Indian nurses. AB - AIM: To identify the factors responsible for the international migration of Indian nurses. METHODS: The paper is based on the responses of 448 nurse practitioners, nurse educators and nurse administrators to a questionnaire administered to them in December 2004-January 2005. Key factors were identified by the analysis of contingency tables. DISCUSSION: Apart from economic factors, dissatisfaction with working conditions and unhappiness with prevalent social attitudes towards nurses were identified as being of crucial importance for the international migration of Indian nurses. It was found that nurses working in the private sector and from some linguistic and religious groups were particularly prone to migration. Nurses working in the government sector seemed to be more worried about being unable to adjust to working conditions abroad, and therefore less keen to migrate. The fact that they enjoyed better pay scales, a more relaxed work atmosphere and more facilities may have also played a part here. What seemed to be vital to the decision to migrate for a large number of government sector nurses belonging to the so-called 'Forward' and 'Middle' Castes was that they were being crowded out of promotional avenues as a result of the government's policy of Reservations in Promotions for Scheduled Castes and Tribes. CONCLUSION: Health policy-makers in India need to take a serious look at the growing migration of nurses to foreign countries. While such migration leads to inflow of foreign exchange, it also implies the loss of medical personnel vital for the fulfilment of national goals. PMID- 17083417 TI - Nursing in the United Arab Emirates: an historical background. AB - AIM: To gain a better understanding of some of the factors that may be contributing to the shortage of national nurses in the United Arab Emirates (UAE). BACKGROUND: Approximately 3% of the UAE nursing workforce are currently nationals, which explains the UAE dependency on transient expatriate nurses. Even though Emirati women have been recently empowered to join the national workforce, not many have joined the nursing profession. Several factors may be contributing, in varying degrees, to this predicament. METHOD: The socio-economic factors as well as the cultural and religious customs that have shaped the country and its people are examined. Hence, the historical background to the establishment of the UAE, women in Islam, the first nurse in Islam, the development of nursing in the UAE and the Emirates Nursing Association are considered. CONCLUSION: Factors contributing to the limited number of UAE nationals in the nursing profession include, but are not limited to: the low status of nursing in the UAE; the variations in basic nursing programmes in the country; the lack of Arabic educational resources; the affluent life style of UAE nationals as well as the strict cultural norms and religious values by which they live. As the need to nationalize the nursing workforce in the UAE is paramount, these issues and other perceived barriers from the perspective of Emirati nurses need to be explored and addressed. PMID- 17083418 TI - Nurse prescribing in low-resource settings: professional considerations. AB - AIM: This paper reviews the professional issues relating to nurse prescribing in low-resource countries. BACKGROUND: In order to meet community healthcare need, nurses in low-resource settings often perform roles such as prescribing medication, for which they may not have had adequate training, often in the absence of supervision, legislation and regulation. In general, however, there is a paucity of published evidence relating to nurse prescribing in low-resource settings. Several studies have demonstrated improved health and social outcomes resulting from nurse-prescribing interventions. Alternatively, other studies have shown unsatisfactory nurse-prescribing patterns, such as overuse of antibiotics and poor adherence to national treatment guidelines. There are several examples of how nurse prescribing in low-resource countries has been recognized and addressed at national level; these are discussed in detail. However, given that it is widely acknowledged how crucial nurses are for the delivery of health care in regions where there are no medical personnel, international policy and guidance relating to this issue is only just beginning to acknowledge and address the relevant issues. CONCLUSION: Where there is a need for nurses to extend their role in the ordering of medicines and other treatments, the responsibilities, training, rights and roles of these nurses need to be clearly defined and recognized at all levels of the health service. There is a need for rigorous evaluations incorporating health, social and economic outcomes of nurse prescribing interventions, in addition to close monitoring of legislative and regulatory changes related to nursing roles. PMID- 17083419 TI - Applying to the European Research Area Network Scheme (ERA-NET): collaborative working for nursing and midwifery research. AB - AIM: This paper reports on work undertaken to achieve an application for monies from the European Commission's 6th Framework Programme by some key stakeholders, working with a nursing and midwifery research agenda at national policy levels. BACKGROUND: A short outline of the European Commission's European Research Area Network scheme is given in order to set the paper in context, and the vision underpinning the application is discussed. CONCLUSION: The paper describes the processes that were undertaken to bring to fruition such collaborative work, and some key lessons are outlined. Seeking opportunities to enhance nursing and midwifery research within a European context gives value to the application, which was ultimately successful. PMID- 17083420 TI - Primary Health Care Nurses' conceptions of involuntarily migrated families' health. AB - BACKGROUND: Involuntary migration and adaptation to a new cultural environment is known to be a factor of psychological stress. Primary Health Care Nurses (PHCNs) frequently interact with refugee families as migrant health needs are mainly managed within Primary Health Care. AIM: To describe the health of the involuntary migrated family in transition as conceptualized by Swedish PHCNs. METHOD: Thirty-four PHCNs from two municipalities in Sweden were interviewed and phenomenographical contextual analysis was used in analysing the data. FINDINGS: Four family profiles were created, each epitomizing the health characteristics of a migrated family in transition: (1) a mentally distressed family wedged in the asylum-seeking process, (2) an insecure family with immigrant status, (3) a family with internal instability and segregated from society, and (4) a stable and well-functioning family integrated in society. Contextual socio-environmental stressors such as living in uncertainty awaiting asylum, having unprocessed traumas, change of family roles, attitudes of the host country and social segregation within society were found to be detrimental to the well-being of the family. CONCLUSION: Acceptance and a clear place in society as well as clearly defined family roles are crucial in facilitating a healthy transition for refugee families. Primary Health Care Nursing can facilitate this by adopting a family system perspective in strengthening the identity of the families and reducing the effects of socio-environmental stressors. PMID- 17083421 TI - Intervention studies for prevention of pressure ulcers in Turkey: a literature review. AB - AIM: To examine critically the literature published in the Turkish language (1955 2004) related to risk assessment tool(s), their application to nursing care and prevention of pressure ulcers (PrUs). Specific objectives were to identify the advantages of risk assessment and prevention of PrUs in hospital; to establish the most valid and reliable methods available to evaluate the effectiveness of PrU prevention programmes; to determine methodological problems encountered by researchers and explore how these were overcome; and to present the findings so they could be used to develop a valid and reliable audit tool based upon the empirical evidence. METHODS: All journals and convention booklets published in Turkey related to nursing between the years 1955-2004 were examined. Because many journals in Turkey are not yet available by electronic means, the published Turkish articles were all examined by hand. A total of 3031 articles in 17 nursing journals and 36 congress books (convention booklets) were examined. Five articles were found to meet the study criteria and were taken into the study and evaluated. CONCLUSION: There is a need to determine valid and reliable assessment methods, and the results need to be recorded on standard forms. In addition, it is important to increase the motivation of nursing personnel who give direct patient care to use the tools available in order to prevent the development of PrUs. From the review findings it is apparent that in Turkey, there is a dearth of research evidence upon which to base practice in the sphere of PrU prevention, and further research is urgently required. PMID- 17083422 TI - An approach to identify dairy cows being responsive to recombinant bovine somatotropin. AB - Forty-eight primiparous and pregnant Holstein cows at 202 (+/-59) days in milk and average daily milk production (DMP) of 22 (+/-5.3) kg at the beginning of the trial were used to study the possibility of identifying animals that are more responsive to recombinant bovine somatotropin (rbST). Animals were housed in free stall barns, milked three times a day and fed a total mixed ration. Animals were injected with one dose (500 mg each) of rbST every 21 days during a 105-day total period (five doses in all). Daily milk production was recorded to measure average apparent response (AAR) and peak of apparent response (PAR). Average apparent response was defined as the average increase in milk production, expressed as kilogram per cow per day, for days 0-18 after dosing, as compared to the average production calculated for days -1, -2, -3, 19, 20 and 21 after dosing. Peak of apparent response was calculated as the increase in milk production for each day from 0 to 18 days after dosing as compared to the average production for days -1, -2 and -3. There was a positive linear correlation (r = 0.608; p = 0.0001) between AAR for the first 21 days and AAR for the total 105-day period (AATR). A correlation coefficient was calculated between each individual PAR value and AATR. The highest correlation coefficient (r = 0.511; p = 0.0002) was observed for days 8, 9 and 10, and therefore these days were considered the best estimates for calculating AAR when DMP records are not available. Also, high somatic cells count had a negative impact on milk yield response to treatment (p = 0.0162). PMID- 17083423 TI - Excess dietary cystine intensifies the adverse effect of a methionine deficiency in the cat. AB - Foot pad dermatitis has been observed in turkeys, puppies and kittens fed diets deficient in methionine. Excess cystine aggravated the lesions and decreased body weight gain in puppies and turkeys. The objective of this study was to determine whether methionine deficiency induced perioral and foot pad lesions in kittens and whether excess cystine exacerbated the lesions. Eighteen kittens were divided into three groups and offered one of three diets: diet 1, low-methionine, low cystine (LMLC; 1.6 g methionine and 1.6 g cystine/kg diet); diet 2, low methionine, high-cystine (HMHC; 1.6 methionine and 15 g cystine/kg diet); diet 3, high-methionine, high-cystine (HMHC; 15 g methionine and 15 g cystine/kg diet). Kittens in the LMLC group lost body weight, whereas those in the LMHC group maintained their body weight and those in the HMHC group gained weight. Plasma methionine concentrations were significantly higher (p < 0.001) for the HMHC group than for the LMLC and LMHC groups. Plasma cyst(e)ine (sum of cysteine and cystine) concentrations were different (p < 0.001) among all the three groups. Two kittens given the LMLC diet developed mild perioral lesions. All kittens receiving the LMHC diet developed foot pad lesions and severe perioral lesions. Histopathological changes observed in perioral biopsy specimens were similar to those described in protein deficiency. In conclusion, the results showed that a diet severely deficient in methionine causes perioral lesions in kittens, and that addition of excess cystine to the diet aggravates the perioral lesions and also causes foot pad lesions. PMID- 17083424 TI - Effect of supplemental yeast culture and sodium bicarbonate on ruminal fermentation and blood variables in rams. AB - A trial was conducted to evaluate the effect of sodium bicarbonate (BC) and Saccharomyces cerevisiae, live yeast culture (yea sacc(1026), YS) on ruminal fermentation and blood variables. Four Kivircik rams with ruminal cannula were used in a Latin square design, during 27-day periods (20 days for adaptation and 7 days for collection). They received 0 (control group), 5 g/day (i.e. 25 x 10(9) CFU) of YS or 10 g/day of sodium BC or 10 g/day of BC and 5 g/day of YS (BC + YS) (treatment groups). The cultures and sodium BC were added to the ration in a grain mix. The ration consisted of 70% grain mix and 30% lucerne hay. Rumen contents were collected before and 3 h and 6 h after morning feeding on days 1 and 7 in each collection period and were analyzed. Blood samples were also collected on the same days. Ruminal pH at 3 h (p < or = 0.1) and 6 h (p < or = 0.05) after feeding were higher when rams were fed BC and BC + YS than when they were fed CG and YS. Addition of YS to the diet did not modify the proportions of the different protozoa types; only Diplodinium at 0 h tended to be lower (p < 0.1) when rams were fed YS, BC and BC + YS than when they were fed CG. Plasma sodium value decreased by YS and BC + YS. Other biochemical and haematological variables were not affected by treatments. Also total volatile fatty acid, NH3-N concentrations and protozoa counts in the ruminal fluid were not affected by treatments. PMID- 17083425 TI - Determination of effective dosage of GH-releasing factor for blood GH responses in mithun (Bos frontalis). AB - A study was undertaken to determine the effective dosage of GH-releasing hormone (GRF) required to produce blood GH response in mithun (Bos frontalis), a semi wild ruminant species. For the purpose, 12 mithuns averaging 11.5 months of age and 146 kg body weight (BW) were randomly assigned to receive GRF (n = 12), administered at 0 (normal saline), 5, 10 and 20 mug per 100 kg BW. Blood samples were collected prior to and after GRF administration at -60, -45, -30, -15, -10, 5, 0 min and 5, 10, 15, 30 and thereafter, at 15-min interval up to 8 h post-GRF were assayed for plasma GH. For all the dosages, the pre-treatment GH concentrations and corresponding area under GH response curve (AUC) were similar (p > 0.05). The post-GRF plasma GH responses to different dosages of GRF viz. 5, 10 and 20 mug per 100 kg BW and corresponding AUCs were higher (p < 0.05) than those recorded in normal saline-treated controls. The GH responses to 10 and 20 mug GRF per 100 kg BW and corresponding AUCs were higher (p < 0.05) than those registered in mithuns administered with 5 mug GRF per 100 kg BW. Interestingly, post-GRF concentration of plasma GH and AUCs were not different for 10 and 20 mug GRF per 100 kg BW dosages. In all animals treated with GRF, a peak of GH was registered within 10 to 20 min post-GRF. Following 5 mug GRF per 100 kg BW, GH concentrations were maintained at higher level for 90 min post-GRF and thereafter became similar to that of controls and it was 435 min for 10 and 20 mug GRF per 100 kg BW dosages. In conclusion, our results suggest that 10 mug GRF per 100 kg BW is the dosage, which can be used for augmentation of mithun production. PMID- 17083426 TI - Effects of copper, zinc and selenium status on performance and health in commercial dairy and beef herds: Retrospective study. AB - A retrospective study using analysis of plasma copper and zinc, and erythrocyte glutathione peroxidase from 2 080 dairy and beef cow herds was conducted to evaluate the relationship between trace-element status and production, reproduction and health in cows and their calves. Classification of the herd status as deficient, marginal, low-adequate or high-adequate was based on the lower tercile of individual values. Odds ratios for each disorder in herds were calculated by multivariable stepwise logistic regression. Inadequate copper status was not associated with adult disorders, but was an important risk factor for poor calf performance or health. Selenium deficient status was associated with most studied disorders in cows, and both deficient and marginal herd status were strongly associated with poor health of calves, particularly with increased risks of myopathy and infectious diseases. Zinc insufficiency was strongly associated with low milk production and impaired locomotion in dairy herds, and was also associated with diarrhoea and poor growth in calves. Because a low adequate status increased the risk of many disorders in adults and calves, we propose to classify herds as deficient and marginal when the lower terciles of plasma zinc concentration are below 12 and between 12 and 14 mumol/l respectively. PMID- 17083427 TI - Effects of dietary neutral detergent fibre on production performance, nutrient utilization, caecum fermentation and fibrolytic activity in 2- to 3-month-old New Zealand rabbits. AB - An experiment was conducted to determine the effects of different dietary neutral detergent fibre (NDF) levels on growth performance, nutrient utilization, caecum development, caecal fermentation, slaughter performance, immune organ and fibrolytic activity in 2- to 3-month-old New Zealand meat rabbits. Eighty 2-month old rabbits were allocated in individual cages for five treatments in which they were fed each diet with NDF at 240, 270, 300, 330 and 360 g/kg respectively. The results are as follows: the average daily gain of 300 g/kg NDF group was the highest (30.09 g/day) and higher than that of 240 g/kg NDF group (p < 0.05). The feed/gain ratio of 300 g/kg NDF group was the lowest (4.27) and lower than those of 240 and 360 g/kg NDF groups (p < 0.05). The coefficient of total tract apparent digestibility (CTTAD) of energy, crude protein, crude fibre and ether extract decreased when dietary NDF increased (p < 0.05); CTTAD of NDF increased when dietary NDF increased (p < 0.01). Digestible nitrogen and retained nitrogen of 240, 270, 300 and 330 g/kg NDF groups were higher than that of 360 g/kg NDF group (p < 0.05). The caecum weight, the proportion of caecum weight to body weight and the acetic acid ratio of volatile fatty acid rose when dietary NDF concentration increased (p < 0.05). The NH(3)-N concentration of caecum dropped when dietary NDF increased (p < 0.05). The fibrolytic activity increased when the dietary NDF increased (p < 0.01). PMID- 17083428 TI - Studies on the tryptophan requirement of lactating sows. Part 1: Estimation of the tryptophan requirement by performance. AB - Mature sows were evenly assigned to six dietary treatments, which differed only in the concentration of total (apparent ileal digestible, AID) tryptophan (Trp) amounting per kg feed to 1.2 g (0.8 g) to 4.2 g (3.8 g) in diets 1-6. For a total of 72 lactations (28 days) sows' feed intake was measured daily and body weight was measured on day 110 of pregnancy and on days 1 and 28 of lactation. Litter size was standardized to 10-12 piglets within 1 week after farrowing. The piglets were weighed at days 1, 21 and 28. Milk yield was recorded by the weigh-suckle weigh method on days 20 and 21. Concentrations of milk fat and protein were measured in manually drawn samples (days 20 and 21) by infrared spectroscopy. Average daily feed intake of the sows was 3.4, 4.7, 4.7, 6.1, 5.1 and 4.7 kg (diets 1-6 respectively, p < 0.001). Body weight losses of the sows during lactation were 30.8, 16.9, 19.2, 9.5, 14.1 and 13.1 kg (diets 1-6 respectively, p < 0.05). Milk output in the average of days 20 and 21 was 6840, 8192, 8362, 9035, 8618 and 8140 g/day, respectively (p < 0.01). There were no differences in milk fat or protein contents. Daily weight gain of the piglets in the average of lactation was 157, 184, 184, 187, 185 and 177 g, respectively (p < 0.10). Based on the results of this study the estimated Trp requirement of lactating sows for optimal performance was calculated with 1.9 g AID Trp/kg in the lactation diet (according to 2.0 g true ileal digestible Trp and approximately 2.6 g Trp on total basis). PMID- 17083429 TI - Studies on the tryptophan requirement of lactating sows. Part 2: Estimation of the tryptophan requirement by physiological criteria. AB - Mature sows were fed for a total of 72 lactations with diets which provided an adequate supply of energy and nutrients except for tryptophan (Trp). By supplementing a basal diet [native 1.2 g Trp/kg, equivalent to 0.8 g apparent ileal digestible (AID) Trp or 0.9 g true ileal digestible (TID) Trp] with L-Trp, five further diets (2-6) containing 1.5-4.2 g Trp/kg were formulated. The dietary Trp content had no effect on amino acid contents in milk on days 20 and 21 of lactation, but Trp in blood plasma on day 28 of lactation reflected the alimentary Trp supply with an increase from 2.74 +/- 1.14 mg/l (diet 1) to 23.91 +/- 7.53 mg/l (diet 6; p < 0.001). There were no directional differences between the diets with regard to the other amino acids. Concentrations of urea in milk and blood were higher with diet 1 (211 and 272 mg/l, respectively) than with diets 3-6 (183 and 227 mg/l, respectively). Serotonin levels in the blood serum were lower with diet 1 (304 ng/ml) than the average of diets 4-6 (540 ng/ml). This study confirms previously given recommendations for the Trp content in the diet of lactating sows, estimated by means of performance, of 1.9 g AID Trp (equivalent to 2.0 g TID Trp; approximately 2.6 g gross Trp) per kg diet. PMID- 17083430 TI - Effects of L-carnitine supplementation in pregnant sows on plasma concentrations of insulin-like growth factors, various hormones and metabolites and chorion characteristics. AB - Previous studies have shown that supplementation of sow diets with L-carnitine increases the body weight of piglets at birth. This study was conducted to elucidate the reasons for this phenomenon. Three experiments with 24 (experiment 1), 40 (experiment 2) and 12 (experiment 3) sows were conducted. In all three experiments, sows were allotted to two groups which had free access to a nutritionally adequate diet. Sows of one group were supplemented with 125 mg L carnitine/day during pregnancy; sows of the other group (control group) did not receive L-carnitine. In experiment 1, plasma samples were collected at day 95 of pregnancy, in experiment 2 plasma samples were collected at days 80 and 100 of pregnancy. In experiment 3, chorions of the sows were collected at parturition. L carnitine-treated sows had higher plasma concentrations of total L-carnitine than control sows (p < 0.05). The number of piglets born and weights of litter and individual piglets at birth were not different between both groups in all three experiments. L-carnitine-treated sows had higher plasma concentrations of insulin like growth factor-I (IGF-I) on day 80 of pregnancy (experiment 2, p < 0.05) and on day 95 (experiment 1, p < 0.10), and a higher plasma concentration of IGF-II on day 80 (experiment 2, p < 0.05) than control sows. Moreover, sows supplemented with L-carnitine had heavier chorions (+22%, p =0.10) with greater amounts of protein (+45%, p < 0.05) and DNA (+38%, p < 0.10) and a higher protein concentration of glucose transporter-1 (+62%, p < 0.05). Plasma concentrations of 17beta-oestradiol, progesterone and thyroid hormones as well as concentrations of urea and total free amino acids were not different between both groups of sows. Plasma concentrations of non-esterified fatty acids, ketone bodies, triacylglycerols and cholesterol were also largely indifferent between both groups of sows. In conclusion, this study shows that L-carnitine has less influence on lipid metabolism and utilization of nitrogen in pregnant sows but increases their plasma concentrations of IGFs. This in turn may enhance development of the placentae and the intrauterine nutrition of the fetuses. This may be the reason for increased birth weights observed in recent studies in sows supplemented with L-carnitine. PMID- 17083431 TI - Effects of essential oils from fennel (Foeniculi aetheroleum) and caraway (Carvi aetheroleum) in pigs. AB - The ban of antibiotics as a feed additive requires alternatives to stabilize the health and performance particularly of the young animals. Essential oils obtained from fennel seed (Foeniculi aetheroleum) and caraway seed (Carvi aetheroleum) were tested in diets for weaned piglets in comparison with either a diet without feed additive or with a combination of formic acid and copper (positive control). Four groups of sixteen piglets (live weight 7 kg, age 26 days) received diets without (1) or with supplements of 7.5 g formic acid + 160 mg Cu/kg (2), 100 mg fennel oil/kg (3) or 100 mg caraway oil/kg (4) during 3 weeks after weaning. In the subsequent 4 weeks, all piglets were fed a diet without these additions. Fennel oil contained almost 2/3 anethol, approximately 1/5 fenchon and the remaining part consisting of alpha + beta-pinen, limonen (p-mentha-1,8-dien) and estragol. In the caraway oil, half of the contents was represented by limonen and the other half by carvon. There were no piglet losses and only few cases of diarrhoea. The combination of formic acid and copper increased feed consumption by 27% and daily weight gain by 25%. There were no differences in the performance between the group fed fennel oil and the control without additives. Piglets fed caraway oil tended to consume less feed and to gain approximately 10% less. In feed choice experiments, pigs consumed the same two diets from two troughs with 50% of total feed amount, as expected. The diets containing fennel or caraway oils were consumed at less than 50%. If the diet contained 100 mg fennel oil/kg, the decrease of percentual feed intake was significant. The results of the feeding experiment and of the feed choice experiment question the classification of fennel and caraway oils as flavour additives or as 'appetite promoters' in diets for weaned piglets. PMID- 17083432 TI - Adaptation of the rumen microbial population to native potato starch degradation determined with the gas production technique and the nylon bag technique. AB - Experiments were conducted to investigate the influence of the adaptation of rumen micro-organisms on the degradation of native potato starch (PS) in the rumen. Cows were fed with rations used for gas production (GP) analysis (dry cows, 1.6% starch) and for the nylon bag (NB) technique (lactating cows, 23% starch, mainly maize starch) and a ration containing 19% native PS (lactating cows). Fermentation characteristics of 13 samples were investigated with the GP technique using rumen fluid from cows fed each of the three rations. The same samples were investigated with the NB technique in the cows obtaining the NB ration and the PS ration. The results showed that the rate of GP was influenced by the source of the rumen fluid. The fermentation rate of PS was considerably enhanced by using rumen fluid adapted to the fermentation of native PS instead of using the other rumen fluids. Incubating in cows fed the PS ration, the rate of PS degradation determined with the NB technique, was higher compared with cows fed other rations. Using the PS ration the observed lag period for PS was shorter. The results show a clear influence of ration on the degradation characteristics of starch, determined with both the GP technique and the NB technique. However, these changes in behaviour did not explain observed differences in amounts of rumen escape PS measured in vivo in animal experiments and in situ, using the NB technique. PMID- 17083433 TI - Disturbing trends in dental education. PMID- 17083434 TI - Longevity of anterior composite restorations. PMID- 17083435 TI - Surgical planning and prosthesis construction using computed tomography, CAD/CAM technology, and the Internet for immediate loading of dental implants. AB - This report describes a protocol that uses computer technology and medical imaging to virtually place anterior and posterior dental implants and to construct a precise surgical template and prosthesis, which is connected at the time of implant placement. This procedure drastically reduces patient office time, surgical treatment time, and the degree of post-treatment recovery. Patients with an edentulous arch or a partially edentulous area had a denture with radiopaque markers constructed for computed tomography (CT) scans of the appropriate jaw. The CT images, having acquisition slices of 0.4 mm, are transposed in a three-dimensional image-based program for planning and strategic placement of dental implants. After virtual implant placement on the computer, the surgical treatment plan is sent to a manufacturing facility for construction of the surgical template. The manufactured surgical components and surgical template arrive on the clinical site. From the surgical template, the dental laboratory retro-engineers the master cast, articulates it with the opposing dentition based on a duplicate of the scanning denture, and creates the prosthesis. Using the surgical template, minimally invasive surgery is performed without a flap, and the prosthesis is delivered, achieving immediate functional loading to the implants. Minor occlusal adjustments are made. The total surgical treatment time required is typically between 30 and 60 minutes. Postoperative symptoms such as pain, swelling, and inflammation are dramatically reduced. CLINICAL SIGNIFICANCE: Identification of the bone in relationship to the tooth position via three-dimensional CT prior to surgery allows the clinician to precisely place implants. Computer-aided design/computer-assisted manufacture technology using the three-dimensional images allows for fabrication of the surgical template. This is a significant advancement in implant dentistry and promotes interdisciplinary approaches to patient treatment. The implant surgeon and restorative dentist can agree upon implant locations and screw access locations prior to the surgical episode. PMID- 17083437 TI - Esthetic potential of single-implant provisional restorations: selection criteria of available alternatives. AB - Provisional implant restorations are essential tools used to optimize the esthetic outcomes for single-tooth implant restorations. This article describes three groups of available alternatives and the specific options within each group including removable prostheses (interim removable partial denture, vacuum-formed appliance), tooth-supported prostheses (bonded extracted or denture teeth, cast metal or fiber-reinforced resin-bonded fixed partial denture [FPD], wire-retained resin-bonded FPD, acrylic resin provisional FPD), and implant-supported fixed prostheses (implant-retained provisional restoration). Advantages and disadvantages as well as fabrication guidelines for each option are provided. The author proposes that the restorative dentist consider eight criteria in selecting the most appropriate type of provisional prosthesis for a specific patient situation including the esthetic potential, patient comfort, treatment time, laboratory cost, occlusal clearance, ease of removal, durability, and ease of modification. The patient's esthetic expectations are critical in determining the most suitable type of provisional restoration. Esthetically pleasing provisional restorations are part of the evolving implant continuum, making implant dentistry more appealing to practicing dentists and potential patients. CLINICAL SIGNIFICANCE: Provisional restorations for single implants have evolved from temporary expedients during osseous and soft tissue integration to critical therapeutic tools used to assess patient expectations, communicate with the laboratory, and optimize definitive implant treatment. The selection of the type of provisional restoration may significantly influence esthetics during the period of implant integration and soft tissue healing. However, it is unlikely that there is a direct correlation between the type of provisional restoration used and the esthetic outcome of the definitive prosthesis. PMID- 17083439 TI - Clinically relevant issues related to preheating composites. AB - STATEMENT OF THE PROBLEM: Issues regarding the use of composite preheating need to be investigated so that the clinician will better understand the variables associated with this method. PURPOSE: To examine the multiple aspects of use of a commercial composite preheating device (Calset, AdDent Inc., Danbury, CT, USA). MATERIALS AND METHODS: Temperature values of three heating units and composite compules were obtained using a K-type thermocouple and were recorded digitally in real time. The following parameters were measured: maximum heater and composite temperature and its stability upon storage, composite temperature change when removed from the heater and injected, the effect of delivery system on ejected composite temperature, and the effect of repeated and extended preheating on composite monomer conversion (using infrared spectroscopy). Monomer conversion was measured after repeated composite cycling (from room temperature [RT] to 60 degrees C, 10x) or extended preheating (24 hours at 60 degrees C), and values were compared with composite maintained at RT (control group). Among test parameters, data (N=5 for each parameter) were analyzed using Student's t-test, analysis of variance, and the Tukey-Kramer post-hoc test where appropriate (alpha=0.05). RESULTS/CONCLUSIONS: Two of the three tested units achieved the stated preset temperatures. Composite attained temperature values close to the heating unit. Composite temperature drop upon removal from the heater was dramatic: within 2 minutes a 50% temperature drop was noted. Heating the compule while preloaded in the syringe provided higher delivery temperatures than heating the compule separately (p < 0.00). Optimum results were achieved when preheated composite was dispensed and used as quickly as possible. Neither repeated nor extended preheating of composite significantly affected monomer conversion. CLINICAL SIGNIFICANCE: Preheating composite has potential benefits, but should be used with knowledge of its limitations. Reheating of unused composite does not affect its degree of conversion, thus decreasing material waste. Heating of the composite preloaded in the delivery syringe enhances the temperature of extruded composite. PMID- 17083441 TI - A survey of edentulous patient preference among different denture esthetic concepts. AB - STATEMENT OF PROBLEM: Despite the fact that solutions to functional problems are available, successfully restoring the appearance of an edentulous patient remains a challenge. PURPOSE: The aim of this survey was to determine if edentulous respondents could differentiate among three denture esthetic concepts and if there was an overall preference among the three. MATERIALS AND METHODS: Six edentulous test subjects were selected based on age and smile criteria. One wax tooth arrangement was completed according to each of the three esthetic concepts for a total of three wax tooth arrangements per test subject. The three esthetic concepts followed were natural, supernormal, and denture look. Standardized full face digital photographs were made of each arrangement (three) during maximum smile for each subject (six). These 18 photographs were included in a questionnaire. Respondents were asked questions about their preference among the three randomly ordered concept photographs for each of the six subjects. A total of 147 completed questionnaires were analyzed. RESULTS: Ninety-six percent of respondents were able to differentiate among the three esthetic denture concepts. Natural tooth arrangements were preferred by 55% of the respondents, supernormal tooth arrangements were preferred by 19%, and the denture look arrangements were preferred by 26%. CONCLUSION: Within the limitations of this survey, the questionnaire respondents differentiated between the three esthetic denture concepts. The tooth arrangement most closely resembling the anatomical average was selected by 55% of the respondents. Preference for a particular concept changed when responses to each test subject set were considered individually. Demographic factors do not significantly affect patient preference. CLINICAL SIGNIFICANCE: Three esthetic concepts for complete denture construction have been differentiated. Questionnaire respondents preferred appearances that are far from the anatomical average 45% of the time. PMID- 17083443 TI - Critical appraisal. Adhesion to laser-prepared tooth structure. PMID- 17083444 TI - Dental adhesives. PMID- 17083445 TI - Somatostatin, a negative-regulator of central leptin action in the rat hypothalamus. AB - Leptin-responsive neurons of the hypothalamus constitute a heterogeneous population expressing a vast array of different neuropeptides and neurotransmitters, some of which participate in the regulation of hunger and satiety. Here we report that somatostatin modulates the efficacy of leptin signalling in the rat hypothalamus. Using a two-pulse paradigm at 30-min intervals, we delivered somatostatin or somatostatin receptor subtype-selective agonists in combination with leptin into the lateral cerebral ventricle of stereotaxically cannulated rats. To monitor the effect of somatostatin on the leptin-signalling pathway, we quantified changes in the leptin-mediated activation of STAT3, the signal transducer and activator of transcription 3. Successive administration of somatostatin and leptin diminished the level of STAT3-phosphorylation and nuclear STAT3 translocation in the ventromedial and dorsomedial hypothalamic nuclei, the lateral hypothalamic area, and the arcuate nucleus by about 40% compared to leptin administration alone. Furthermore, application of subtype-selective somatostatin receptor agonists suggests that the observed reduction in leptin-responsiveness is predominantly mediated by the sst3 receptor-subtype, followed by sst1 and sst2. In addition, the intensity of the negative-regulatory effect of somatostatin on leptin-signalling displayed regional differences for the three receptor-subtypes involved. Addressing the functional consequences of the diminished leptin-signalling, behavioural analyses showed that centrally applied somatostatin counteracts the leptin-mediated suppression of food intake. These results suggest that the pleiotropic effector somatostatin also plays a role in the central regulation of energy homeostasis. PMID- 17083446 TI - GABAA receptor associated proteins: a key factor regulating GABAA receptor function. AB - gamma-Aminobutyric acid (GABA), an important inhibitory neurotransmitter in both vertebrates and invertebrates, acts on GABA receptors that are ubiquitously expressed in the CNS. GABA(A) receptors also represent a major site of action of clinically relevant drugs, such as benzodiazepines, barbiturates, ethanol, and general anesthetics. It has been shown that the intracellular M3-M4 loop of GABA(A) receptors plays an important role in regulating GABA(A) receptor function. Therefore, studies of the function of receptor intracellular loop associated proteins become important for understanding mechanisms of regulating receptor activity. Recently, several labs have used the yeast two-hybrid assay to identify proteins interacting with GABA(A) receptors, for example, the interaction of GABA(A) receptor associated protein (GABARAP) and Golgi-specific DHHC zinc finger protein (GODZ) with gamma subunits, PRIP, phospholipase C related, catalytically inactive proteins (PRIP-1) and (PRIP-2) with GABARAP and receptor gamma2 and beta subunits, Plic-1 with some alpha and beta subunits, radixin with the alpha5 subunit, HAP1 with the beta1 subunit, GABA(A) receptor interacting factor-1 (GRIF-1) with the beta2 subunit, and brefeldin A-inhibited GDP/GTP exchange factor 2 (BIG2) with the beta3 subunit. These proteins have been shown to play important roles in modulating the activities of GABA(A) receptors ranging from enhancing trafficking, to stabilizing surface and internalized receptors, to regulating modification of GABA(A) receptors. This article reviews the current studies of GABA(A) receptor intracellular loop-associated proteins. PMID- 17083447 TI - BACE1 inhibition reduces endogenous Abeta and alters APP processing in wild-type mice. AB - Accumulation of amyloid beta peptide (Abeta) in brain is a hallmark of Alzheimer's disease (AD). Inhibition of beta-site amyloid precursor protein (APP) cleaving enzyme-1 (BACE1), the enzyme that initiates Abeta production, and other Abeta-lowering strategies are commonly tested in transgenic mice overexpressing mutant APP. However, sporadic AD cases, which represent the majority of AD patients, are free from the mutation and do not necessarily have overproduction of APP. In addition, the commonly used Swedish mutant APP alters APP cleavage. Therefore, testing Abeta-lowering strategies in transgenic mice may not be optimal. In this study, we investigated the impact of BACE1 inhibition in non transgenic mice with physiologically relevant APP expression. Existing Abeta ELISAs are either relatively insensitive to mouse Abeta or not specific to full length Abeta. A newly developed ELISA detected a significant reduction of full length soluble Abeta 1-40 in mice with the BACE1 homozygous gene deletion or BACE1 inhibitor treatment, while the level of x-40 Abeta was moderately reduced due to detection of non-full-length Abeta and compensatory activation of alpha secretase. These results confirmed the feasibility of Abeta reduction through BACE1 inhibition under physiological conditions. Studies using our new ELISA in non-transgenic mice provide more accurate evaluation of Abeta-reducing strategies than was previously feasible. PMID- 17083448 TI - Pharmacological approaches towards rationalizing the use of endoparasitic drugs in small animals. AB - Parasitic diseases are an important health concern to small animal veterinarians worldwide, and their zoonotic potential is also of relevance to human medicine. The treatment and control of such conditions relies heavily on pharmaceutical intervention using a range of antiparasitic drugs and/or their biologically active metabolites. Broad spectrum agents have been produced, although narrow and even monospecific drugs are used in some situations. Their efficacy may depend on dosage, the target pathogen(s), the host species and/or the site of infection. Optimal use of antiparasitics requires a detailed consideration of the pharmacokinetic and pharmacodynamic properties of the drugs in specific clinical contexts. This review summarizes the present status of knowledge on the metabolism, and physicochemical and pharmacological properties of the major antiparasitic drugs currently used in small animal veterinary practice. In addition, data relevant to therapeutic dosage, efficacy and clinical indication/contraindication, particularly in relation to combination drug therapy, are included. PMID- 17083449 TI - Comparison of bovine in vivo bioavailability of two sulfamethazine oral boluses exhibiting different in vitro dissolution profiles. AB - The bolus (or oblet) is a dosage form that can be used for the oral administration of pharmaceutical compounds to ruminating species. Unlike traditional tablets, oral boluses may contain quantities of drug on the order of grams rather than milligrams. Due to its size, it is only recently that USP-like in vitro dissolution methods have been developed for this dosage form. However, whether or not these dissolution tests can predict product in vivo performance has yet to be determined. The importance of this issue is apparent when the U.S. Food and Drug Administration Center for Veterinary Medicine is faced with the decision of whether to require additional in vivo bioequivalence study data to support the approval of changes in product chemistry or manufacturing method. The current study was undertaken to determine whether an in vivo/in vitro correlation can be established for bovine sulfamethazine oral boluses and to acquire insight into the magnitude of changes in in vitro product performance that can occur before corresponding changes are seen in in vivo blood level profiles. Based upon the results of this investigation, it is concluded that marked changes in in vitro sulfamethazine bolus performance can be tolerated before resulting in altered in vivo blood level profiles. However, the data also suggest that rumenal absorption may occur for some compounds. Therefore the degree to which variation in product in vitro dissolution profiles can be tolerated may be compound specific. PMID- 17083450 TI - Cytochrome P450-dependent metabolism of midazolam in hepatic microsomes from chickens, turkeys, pheasant and bobwhite quail. AB - In vitro putative cytochrome P450 3A mediated activity, and inhibition thereof, were measured in four avian species using midazolam (MDZ) as a substrate and ketoconazole as an inhibitor. All species produced 1-hydroxymidazolam (1-OH MDZ) to a much greater extent than 4-hydroxymidazolam (4-OH MDZ). Calculated Vmaxapparent values for formation of 1-OH MDZ were 117+/-17, 239+/-108, 437+/ 168, and 201+/-55 pmol/mg protein*min and Kmapparent values were 2.1+/-0.8, 2.4+/ 1.6, 6.7+/-5.1 and 3.2+/-2.1 microm for chicken, turkey, pheasant and bobwhite quail, respectively. For the formation of 4-OH MDZ the Vmaxapparent values were 21+/-10, 94+/-46, 144+/-112, and 68+/-30 pmol/mg protein*min and Kmapparent values for 4-OH MDZ formation were 12.4+/-10.1, 18.0+/-10.8, 38.6+/-34.7 and 29.1+/-10.1 microm for chicken, turkey, pheasant and bobwhite quail, respectively. In all four species, ketoconazole inhibited the production of both major metabolites of MDZ, with 4-OH MDZ formation more sensitive to inhibition than 1-OH MDZ. Pheasant and bobwhite quail appeared most sensitive to ketoconazole inhibition. PMID- 17083451 TI - Pharmacodynamic effects and pharmacokinetic profile of a long-term continuous rate infusion of racemic ketamine in healthy conscious horses. AB - Ketamine (KET) possesses analgesic and anti-inflammatory activity at sub anesthetic doses, suggesting a benefit of long-term KET treatment in horses suffering from pain, inflammatory tissue injury and/or endotoxemia. However, data describing the pharmacodynamic effects and safety of constant rate infusion (CRI) of KET and its pharmacokinetic profile in nonpremedicated horses are missing. Therefore, we administered to six healthy horses a CRI of 1.5 mg/kg/h KET over 320 min following initial drug loading. Cardiopulmonary parameters, arterial blood gases, glucose, lactate, cortisol, insulin, nonesterified fatty acids, and muscle enzyme levels were measured, as were plasma concentrations of KET and its metabolites using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Levels of sedation and muscle tension were scored. Respiration and heart rate significantly increased during the early infusion phase. Glucose and cortisol significantly varied both during and after infusion. During CRI all horses scored 0 on sedation. All but one horse scored 0 on muscle tension, with one mare scoring 1. All other parameters remained within or close to physiological limits without significant changes from pre-CRI values. The mean plasma concentration of KET during the 1.5 mg/kg/h KET CRI was 235 ng/mL. The decline of its plasma concentration-time curve of both KET and norketamine (NKET) following the CRI was described by a two-compartmental model. The metabolic cascade of KET was NKET, hydroxynorketamine (HNK), and 5,6-dehydronorketamine (DHNK). The KET median elimination half-lives (t1/2alpha and t1/2beta) were 2.3 and 67.4 min, respectively. The area under the KET plasma concentration-time curve (AUC), elimination was 76.0 microg.min/mL. Volumes of C1 and C2 were 0.24 and 0.79 L/kg, respectively. It was concluded that a KET CRI of 1.5 mg/kg/h can safely be administered to healthy conscious horses for at least 6 h, although a slight modification of the initial infusion rate regimen may be indicated. Furthermore, in the horse KET undergoes very rapid biotransformation to NKET and HNK and DHNK were the major terminal metabolites. PMID- 17083452 TI - Fumagillin, a new P-glycoprotein-interfering agent able to modulate moxidectin efflux in rat hepatocytes. AB - We have tested the ability of two compounds licensed in veterinary medicine: fumagillin and diminazene diaceturate to increase intracellular moxidectin quantity in rat hepatocytes. These compounds significantly increased the quantity of 14C-moxidectin (expressed as area under the time curve concentrations) in cultured rat hepatocytes by 44% and 65% for diminazene and fumagillin treatments respectively. In addition, we have tested these drugs for their interference with P-glycoprotein (P-gp) function in porcine kidney epithelial cells transfected with murine mdr1a (Mdr1a-LLCPK1). We examined the intracellular accumulation of rhodamine 123 (Rho 123) as a functional test to evaluate the effects of these two drugs on P-gp activity. In this model, only fumagillin led to a marked intracellular accumulation of Rho 123. After transforming the data to express the results as a percentage of the accumulation in the presence of the P-gp inhibitor valspodar (VSP), the maximal Rho 123 accumulation was 47% of that with VSP for 100 microm fumagillin. The EC50, the concentration needed to determine 50% of the maximal effect was 34 microm. Fumagillin interacts with P-gp function and appears as a promising compound among registered drugs available, which may optimize the therapeutic use of macrocyclic lactones (MLs). PMID- 17083453 TI - The disposition of lidocaine during a 12-hour intravenous infusion to postoperative horses. AB - Lidocaine is administered as an intravenous infusion to horses for a variety of reasons, but no study has assessed plasma lidocaine concentrations during a 12-h infusion to horses. The purpose of this study was to evaluate the plasma concentrations and pharmacokinetics of lidocaine during a 12-h infusion to postoperative horses. A second purpose of the study was to evaluate the in vitro plasma protein binding of lidocaine in equine plasma. Lidocaine hydrochloride was administered as a loading dose, 1.3 mg/kg over 15 min, then by a constant rate IV infusion, 50 microg/kg/min to six postoperative horses. Lidocaine plasma concentrations were measured by a validated high-pressure liquid chromatography method. One horse experienced tremors and collapsed 5.5 h into the study. The range of plasma concentrations during the infusion was 1.21-3.13 microg/mL. Lidocaine plasma concentrations were significantly increased at 0.5, 4, 6, 8, 10 and 12 h compared with 1, 2 and 3 h. The in vitro protein binding of lidocaine in equine plasma at 2 microg/mL was 53.06+/-10.28% and decreased to 27.33+/-9.72% and 29.52+/-6.44% when in combination with ceftiofur or the combination of ceftiofur and flunixin, respectively. In conclusion, a lower lidocaine infusion rate may need to be administered to horses on long-term lidocaine infusions. The in vitro protein binding of lidocaine is moderate in equine plasma, but highly protein bound drugs may displace lidocaine increasing unbound concentrations and the risk of lidocaine toxicity. PMID- 17083454 TI - Pharmacokinetics and pharmacodynamics of cefovecin in dogs. AB - A series of in vivo, ex vivo and in vitro studies were conducted to determine the pharmacokinetic and pharmacodynamic properties of cefovecin, a new injectable cephalosporin, in dogs. Absolute bioavailability was determined in a two-phase cross-over study in dogs receiving 8 mg/kg bodyweight (b.w.) of cefovecin by either subcutaneous (s.c.) or intravenous (i.v.) route. After s.c. administration, cefovecin was fully bioavailable (100%), the mean maximum plasma concentration (Cmax) was 121 microg/mL and the mean apparent elimination half life (t1/2) was 133 h. Clearance was measured to be 0.76 mL/h/kg after i.v. dosing. The concentration of cefovecin in urine measured 14 days after s.c. administration was 2.9 microg/mL. Plasma protein binding was determined by equilibrium dialysis; over concentrations ranging from 10 to 100 microg/mL (i.e. up to the approximate Cmax following an 8 mg/kg dose), protein binding of 98.7% to 96.0% was observed, however, binding was lower at higher concentrations. Total and free concentrations of cefovecin were determined in plasma, transudate and exudate collected from dogs previously implanted subcutaneously with tissue cages. Mean peak concentrations of free cefovecin were almost three times higher in transudate than in plasma and remained above 0.25 microg/mL for 19 days. The ex vivo antibacterial killing activity (vs. Staphylococcus intermedius, MIC 0.25 microg/mL) was measured in serum, transudate and exudate collected from dogs which had received 8 mg/kg b.w. of cefovecin subcutaneously. Transudate exhibited higher antimicrobial killing activity than serum. Activity in serum and exudate exhibited a mean reduction in bacterial counts of S. intermedius of at least three log units up to 72 h postadministration. Bactericidal activity (>3 log10 reduction of bacterial counts) was observed in transudate up to 12 days postadministration. The slow elimination and long lasting ex vivo antibacterial killing activity following administration of cefovecin are desirable pharmacokinetic and pharmacodynamic attributes for an antimicrobial drug with 14 day dosing intervals. PMID- 17083455 TI - Pharmacokinetics of cefovecin in cats. AB - The pharmacokinetics of the novel cephalosporin cefovecin were investigated in a series of in vivo, ex vivo and in vitro studies following administration to adult cats at 8 mg/kg bodyweight. Bioavailability and pharmacokinetic parameters were determined in a cross-over study after intravenous (i.v.) and subcutaneous (s.c.) injections. [14C]cefovecin was used to evaluate excretion for 21 days after s.c. administration. Protein binding was determined in vitro in feline plasma and ex vivo in transudate from cats surgically implanted with tissue chambers. After s.c. administration, cefovecin was characterized by rapid absorption with mean peak plasma concentrations of 141+/-12 microg/mL being achieved within 2 h of s.c. injection with full bioavailability (99%). The mean elimination half-life was 166+/-18 h. After i.v. administration, volume of distribution was 0.09+/-0.01 L/kg and mean plasma clearance was 0.35+/-0.04 mL/h/kg. Approximately 50% of the administered radiolabelled dose was eliminated over the 21-day postdose period via urinary excretion and up to approximately 25% in faeces. In vitro and ex vivo plasma protein binding ranged from 99.8% to 99.5% over the plasma concentration range 10-100 microg/mL. Ex vivo protein binding in transudate was as low as 90.7%. From 8 h postdose, concentrations of unbound (free) cefovecin in transudate were consistently higher than in plasma, with mean unbound cefovecin concentrations being maintained above 0.06 microg/mL (MIC90 of Pasteurella multocida) in transudate for at least 14 days postdose. The slow elimination and long-lasting free concentrations in extracellular fluid are desirable pharmacokinetic attributes for an antimicrobial with a 14-day dosing interval. PMID- 17083456 TI - Metabolism and depletion of albendazole in the muscle tissue of channel catfish following oral treatment. AB - The residue depletion of albendazole (ABZ) and its metabolites was studied in channel catfish muscle tissue. Channel catfish were dosed once with 10 mg/kg ABZ via stomach tube with manual restraint. Muscle tissue samples were collected at 8, 16, 24, 48, 72, 96 and 120 h postdose. A high-performance liquid chromatographic method was used to assay ABZ and its major metabolites: ABZ sulfoxide (ABZ-SO), ABZ sulfone (ABZ-SO2) and ABZ aminosulfone (ABZ-2-NH2SO2) in the muscle tissue. The results indicate that ABZ and ABZ-SO were present in low concentrations, i.e. <15 and <10 microg/kg, respectively, at 8 h postdose in catfish muscle with and without skin. ABZ-SO2 was present at 1 microg/kg concentration levels until 48 h in muscle alone and 72 h in muscle with skin. ABZ 2-NH2SO2 was not detected at any withdrawal periods. PMID- 17083457 TI - Effects of subcutaneous methadone, morphine, buprenorphine or saline on thermal and pressure thresholds in cats. AB - This study compared pressure and thermal thresholds after administration of three opioids in eight cats. Pressure stimulation was performed via a bracelet taped around the forearm. Three ball-bearings were advanced against the forearm by inflation of a modified blood pressure bladder. Pressure in the cuff was recorded at the end point (leg shake and head turn). Thermal threshold was tested as previously reported using a heated probe held against the thorax [Dixon et al. (2002) Research in Veterinary Science, 72, 205]. After baseline recordings, each cat received subcutaneous methadone 0.2 mg/kg, morphine 0.2 mg/kg, buprenorphine 0.02 mg/kg or saline 0.3 mL in a four period cross-over study. Measurements were made at 15, 30, 45 min and 1, 2, 3, 4, 8, 12 and 24 h after the injection. Data were analysed by anova (P<0.05). There were no significant changes in thresholds after saline. Thermal threshold increased at 45 min after buprenorphine (maximum 2.8+/-3 degrees C), 1-3 h after methadone (maximum 3.4+/-1.9 degrees C) and 45 min to 1 h (maximum 3.4+/-2 degrees C) after morphine. Pressure threshold increased 30-45 min (maximum 238+/-206 mmHg) after buprenorphine, 45-60 min after methadone (maximum 255+/-232 mmHg) and 45-60 min and 3-6 h (maximum 255+/-232 mmHg) after morphine. Morphine provided the best analgesia, and methadone appears a promising alternative. Buprenorphines limited effect was probably related to the subcutaneous route of administration. Previously, buprenorphine has produced much greater effects when given by other routes. PMID- 17083458 TI - Pharmacokinetics of fentanyl delivered transdermally in healthy adult horses- variability among horses and its clinical implications. AB - The safety and pharmacokinetics of fentanyl, delivered transdermally at a dosage of 60-67 microg/kg, were investigated in six healthy adult horses. Three transdermal fentanyl patches (Duragesic), each containing 10 mg of fentanyl citrate, were applied to the mid-dorsal thorax of each horse and left in place for 72 h. Plasma fentanyl concentrations were periodically measured throughout this period and for 12 h after patch removal. After an initial delay of approximately 2 h, the plasma fentanyl concentration rose rapidly in a fairly linear fashion, reaching a peak at around 12 h; thereafter, it gradually declined in a roughly linear manner over the next 72 h. There was much individual variation, however. The initial delay ranged from 0 to 5.1 h (mean, 1.91+/-2.0 h), Tcmax ranged from 8.5 to 14.5 h (mean, 11.4+/-2.7 h) and Cmax ranged from 0.67 to 5.12 ng/mL (mean, 2.77+/-1.92 ng/mL). In two horses, the plasma fentanyl concentration failed to reach even 1 ng/mL, whereas in the other four horses it was >1 ng/mL for at least 40 h and for at least 72 h in two of these horses. No adverse effects attributable to fentanyl were observed in any of the horses, indicating that this dosage is safe in systemically healthy adult horses. However, it failed to achieve plasma fentanyl concentrations generally considered to be analgesic (>or=1 ng/mL) in about one-third of horses. PMID- 17083459 TI - Metabolism of isometamidium in hepatocytes isolated from control and inducer treated rats. AB - Little is known about the metabolism and mechanism of action of the trypanocide, isometamidium (ISM), the major drug used for prophylaxis of trypanosomiasis. We have investigated its metabolism and distribution in isolated rat hepatocytes using liquid chromatography-mass spectrometry and confocal laser scanning microscopy (CLSM). Two putative metabolites were formed, which were proposed to be a mono-acetyl derivative and an oxidized metabolite (SII). This is the first demonstration of the hepatic metabolism of ISM, as previous in vivo studies were hampered by dose-limiting toxicity and insensitive analytical methods. The intrinsic fluorescence of the drug enabled its intracellular uptake to be followed by CLSM. It is taken up rapidly into the nucleolus, nuclear membrane and endoplasmic reticulum within 5 min, and retained in the nucleus for at least 24 h. Persistent binding of ISM to cellular macromolecules may contribute to its prophylactic effect in vivo. Pretreatment of rats with 3-methylcholanthrene, phenobarbitone (PB) or the widely used pyrethroid pesticide, deltamethrin, resulted in an increase in metabolism of ISM to the proposed SII after 1 h incubation with hepatocytes. 3-methylcholanthrene was the most potent inducer, causing a maximal 19.5-fold induction of SII formation after exposure of hepatocytes to ISM for 1 h compared with formation by control hepatocytes. In comparison, at the 1 h timepoint deltamethrin pre-treatment caused a 10.2-fold induction, and PB only 8.2 fold. PMID- 17083460 TI - Clinical efficacy of intravenous administration of marbofloxacin in a Staphylococcus aureus infection in tissue cages in ponies. AB - Tissue cages (TC), implanted subcutaneously in the neck in eight ponies, were inoculated with Staphylococcus aureus (S. aureus) to determine the clinical efficacy of marbofloxacin in the treatment of this infection. From 21 h after inoculation, marbofloxacin (6 mg/kg) was administered intravenously (i.v.) once daily for 7 days. Samples of the tissue cage fluid (TCF) were taken to determine marbofloxacin concentrations (days 1, 3 and 7), using high-pressure liquid chromatography, and numbers of viable bacteria [colony forming units (CFU)] (days 1, 3, 7, 14 and 21). Statistical analysis was used to compare CFU before and after treatment. Clinical signs and CFU were used to evaluate the efficacy of treatment. Although, there was a slight decrease in CFU in all TC initially, the infection was not eliminated by marbofloxacin treatment in any of the ponies and abscesses formed. As the MIC (0.25 microg/mL) did not change during treatment and the concentration of marbofloxacin during treatment (mean concentration in TCF was 0.89 microg/mL on day 1, 0.80 microg/mL on day 3 and 2.77 microg/mL on day 7) was above MIC, we consider that the treatment failure might be attributable to the formation of a biofilm by S. aureus. Based on the present results, i.v. administration of marbofloxacin alone is not suitable for the elimination of S. aureus infections from secluded sites. PMID- 17083461 TI - Pulmonary disposition of tilmicosin in foals and in vitro activity against Rhodococcus equi and other common equine bacterial pathogens. AB - The objectives of this study were to determine the serum and pulmonary disposition of tilmicosin in foals and to investigate the in vitro activity of the drug against Rhodococcus equi and other common bacterial pathogens of horses. A single dose of a new fatty acid salt formulation of tilmicosin (10 mg/kg of body weight) was administered to seven healthy 5- to 8-week-old foals by the intramuscular route. Concentrations of tilmicosin were measured in serum, lung tissue, pulmonary epithelial lining fluid (PELF), bronchoalveolar lavage (BAL) cells, and blood neutrophils. Mean peak tilmicosin concentrations were significantly different between sampling sites with highest concentrations measured in blood neutrophils (66.01+/-15.97 microg/mL) followed by BAL cells (20.1+/-5.1 microg/mL), PELF (2.91+/-1.15 microg/mL), lung tissue (1.90+/-0.65 microg/mL), and serum (0.19+/-0.09 microg/mL). Harmonic mean terminal half-life in lung tissue (193.3 h) was significantly longer than that of PELF (73.3 h), bronchoalveolar cells (62.2 h), neutrophils (47.9 h), and serum (18.4 h). The MIC90 of 56 R. equi isolates was 32 microg/mL. Tilmicosin was active in vitro against most streptococci, Staphylococcus spp., Actinobacillus spp., and Pasteurella spp. The drug was not active against Enterococcus spp., Pseudomonas spp., and Enterobacteriaceae. PMID- 17083462 TI - Nonsteroidal anti-inflammatory drugs induce hypermotilinemia and disturbance of interdigestive migrating contractions in instrumented dogs. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) induce gastric ulcers due to inhibition of prostaglandin production. Prostaglandins have an influence on physiological gastrointestinal motility, but the relationships between NSAID induced gastric ulcer, gastrointestinal motility and motilin are unknown. Fifteen dogs were allocated randomly to three groups in which either gelatin, meloxicam or indomethacin was administered. Fecal occult blood and gastrointestinal motility were monitored continuously for 6 days. In addition, analyses of the plasma motilin concentration, gastrointestinal endoscopy and gastric emptying, and detection of motilin cells were performed. Gastrointestinal motility was disturbed in the indomethacin group, presenting as disappearance of interdigestive migrating contractions (IMCs) 3 days before gastric ulcers were detected. Delayed gastric emptying and hypermotilinemia were observed significantly more often in the indomethacin group compared with the other groups. Motilin cell-crypt/villi ratio in the indomethacin group significantly decreased in the duodenum and jejunum, compared with the other groups. No significant changes in any tests were observed in the meloxicam group, when compared with the gelatin group. These findings suggest that the disturbance of IMCs caused by hypermotilinemia, with changes in motilin cell distribution, and delayed gastric emptying induced by indomethacin may contribute to the development of gastric ulcers. PMID- 17083463 TI - Determination of plasma protein binding of diltiazem in horses by ultrafiltration. PMID- 17083464 TI - Dose selection and pharmacokinetics of rifampin in elephants for the treatment of tuberculosis. PMID- 17083465 TI - Pharmacokinetics of tiletamine and zolazepam (Telazol) in anesthetized pigs. PMID- 17083466 TI - Identification of an essential gene responsible for D-Asp incorporation in the Lactococcus lactis peptidoglycan crossbridge. AB - Bacteria such as Lactococcus lactis have D-aspartate (D-Asp) or its amidated derivative D-asparagine (D-Asn), in their peptidoglycan (PG) interpeptide crossbridge. We performed a subtractive genome analysis to identify L. lactis gene yxbA, orthologues of which being present only in bacteria containing D-amino acids in their PG crossbridge, but absent from those that instead insert L-amino acids or glycine. Inactivation of yxbA required a complementing Streptococcus pneumoniae murMN genes, which express enzymes that incorporate L-Ser-L-Ala or L Ala-L-Ala in the PG crossbridge. Our results show that (i) yxbA encodes D-Asp ligase responsible for incorporation of D-Asp in the PG crossbridge, and we therefore renamed it as aslA, (ii) it is an essential gene, which makes its product a potential target for specific antimicrobials, (iii) the absence of D Asp may be complemented by L-Ser-L-Ala or L-Ala-L-Ala in the L. lactis PG, indicating that the PG synthesis machinery is not selective for the side-chain residues, and (iv) lactococcal strains having L-amino acids in their PG crossbridge display defects in cell wall integrity, but are able to efficiently anchor cell wall proteins, indicating relative flexibility of lactococcal transpeptidation reactions with respect to changes in PG sidechain composition. PMID- 17083467 TI - A peptidoglycan hydrolase motif within the mycobacteriophage TM4 tape measure protein promotes efficient infection of stationary phase cells. AB - The predominant morphotype of mycobacteriophage virions has a DNA-containing capsid attached to a long flexible non-contractile tail, features characteristic of the Siphoviridae. Within these phage genomes the tape measure protein (tmp) gene can be readily identified due to the well-established relationship between the length of the gene and the length of the phage tail--because these phages typically have long tails, the tmp gene is usually the largest gene in the genome. Many of these mycobacteriophage Tmp's contain small motifs with sequence similarity to host proteins. One of these motifs (motif 1) corresponds to the Rpf proteins that have lysozyme activity and function to stimulate growth of dormant bacteria, while the others (motifs 2 and 3) are related to proteins of unknown function, although some of the related proteins of the host are predicted to be involved in cell wall catabolism. We show here that motif 3-containing proteins have peptidoglycan-hydrolysing activity and that while this activity is not required for phage viability, it facilitates efficient infection and DNA injection into stationary phase cells. Tmp's of mycobacteriophages may thus have acquired these motifs in order to avoid a selective disadvantage that results from changes in peptidoglycan in non-growing cells. PMID- 17083468 TI - MvfR, a key Pseudomonas aeruginosa pathogenicity LTTR-class regulatory protein, has dual ligands. AB - MvfR (PqsR), a Pseudomonas aeruginosa LysR-type transcriptional regulator, plays a critical role in the virulence of this pathogen. MvfR modulates the expression of multiple quorum sensing (QS)-regulated virulence factors; and the expression of the phnAB and pqsA-E genes that encode functions mediating 4-hydroxy-2 alkylquinolines (HAQs) signalling compounds biosynthesis, including 3,4-dihydroxy 2heptylquinoline (PQS) and its precursor 4-hydroxy-2-heptylquinoline (HHQ). PQS enhances the in vitro DNA-binding affinity of MvfR to the pqsA-E promoter, to suggest it might function as the in vivo MvfR ligand. Here we identify a novel MvfR ligand, as we show that HHQ binds to the MvfR ligand-binding-domain and potentiates MvfR binding to the pqsA-E promoter leading to transcriptional activation of pqsA-E genes. We show that HHQ is highly produced in vivo, where it is not fully converted into PQS, and demonstrate that it is required for MvfR dependent gene expression and pathogenicity; PQS is fully dispensable, as pqsH mutant cells, which produce HHI but completely lack PQS, display normal MvfR dependent gene expression and virulence. Conversely, PQS is required for full production of pyocyanin. These results uncover a novel biological role for HHQ; and provide novel insights on MvfR activation that may aid in the development of therapies that prevent or treat P. aeruginosa infections in humans. PMID- 17083469 TI - Proteins encoded by the conservon of Streptomyces coelicolor A3(2) comprise a membrane-associated heterocomplex that resembles eukaryotic G protein-coupled regulatory system. AB - Streptomyces coelicolor A3(2) retains unique conserved operons termed conservons. Here, one of the conservons (cvn9), which encodes five proteins (A9-E9), was characterized. Mutants for cvnA9 and cvnAlO conditionally overproduced actinorhodin and performed precocious aerial growth, while a cvnE9 mutant showed the parental phenotype. Transcription of bidG, adpA and bldN was upregulated in the cvnA9 mutant. A9-D9 were detected in the insoluble fraction of cell-free extract of S. coelicolor by Western analysis. Biochemical analyses revealed that A9 has ATP-hydrolysing and adenine nucleotide-binding activities; D9 has GTP hydrolysing and guanine nucleotide-binding activities; and E9 shows a typical spectrum similar to cytochrome P450. The comprehensive interaction assays demonstrated the occurrence of specific interactions between A9 and B9, A9 and C9, B9 and B9, B9 and D9, and C9 and D9. A9 associated with and dissociated from B9 (and C9) when ATP and ATP-gamma-S were supplied in the reaction respectively. Similarly, D9 associated with and dissociated from B9 (and C9) when GTP and GTP gamma-S were supplied respectively. A9 and B9 were also shown for the occurrence as homocomplexes. Probably, Cvn9 proteins comprise a membrane-associated heterocomplex resembling the eukaryotic G-protein-coupled receptor system, which may serve as a signal transducer that connects to the bld cascade. PMID- 17083470 TI - Functional coupling between the two active sites during Tn 10 transposition buffers the mutation of sequences critical for DNA hairpin processing. AB - DNA processing reactions often involve multiple components acting in concert to achieve the desired outcome. However, it is usually difficult to know how the components communicate and cooperate to orchestrate an ordered series of events. We address this question in the context of the Tn 10 transposition reaction, in which the DNA cleavage and joining events occur within a higher-order complex containing a transposase dimer, two transposon ends and the DNA-bending host factor IHF (Integration Host Factor). Previously it was shown that the complex is asymmetric. The a side consists of an IHF protomer initially immobilized by a DNA loop, but subsequently used to promote conformational changes required for the cleavage steps. The beta side of the complex was considered to fulfil a more passive role. Here we show that the a side of the complex promotes coupled conformational changes at both transposon ends, while the a and beta sides communicate and cooperate to dominate different phases of the transposition reaction. Together, these effects provide for a robust response to critical changes in the transposon end. These findings also explain the intriguing genetic phenotypes of a series of previously reported Tn10 mutants and have consequences for the evolution of new elements. PMID- 17083471 TI - Are cranial germ cell tumours really tumours of germ cells? AB - Germ cell tumours of the brain and those that occur in the gonads are believed to share a common origin from germ cell progenitors. This 'germ cell theory' rests upon similar histopathology between these tumours in different locations and the belief that endogenous somatic cells of the brain could not give rise to the range of cell types seen in germ cell tumours. An alternative 'embryonic cell theory' has been proposed for some classes of cranial germ cell tumours, but this still relies on the misplacement of cells in the brain (in this case the earliest embryonic stem cells) during early embryonic development. Recent evidence has demonstrated that neural stem cells of the brain can also give rise to many of the cell types seen in germ cell tumours. These data suggest that endogenous progenitor cells of the brain are a plausible alternative origin for these tumours. This idea is of central importance for studies aiming to elucidate the mechanisms of tumour development. The application of modern molecular analyses to reveal how tumour cells have altered with respect to their cell of origin relies on the certain identification of the cell from which the particular tumour arose. If the identity of this cell is mistaken, then studies to elucidate the mechanisms by which the progenitor cell has been subverted from its normal behaviour will not yield useful information. In addition, it will prove impossible to generate an appropriate animal model in which to study the underlying causes of those tumours. This article makes the case that current assumptions of the origins of cranial germ cell tumours are unreliable. It reviews the evidence in favour of the 'germ cell theory' and argues in favour of a 'brain cell theory' in which endogenous neural progenitor cells of the brain are the likely origin for these tumours. Thus, the case is made that cranial germ cell tumours, like other brain tumours, arise by the transformation of progenitor cells normally resident in the brain. PMID- 17083472 TI - The in vitro influences of neurotensin on the motility characteristics of human U373 glioblastoma cells. AB - Astrocytic tumours are associated with dismal prognoses due to their pronounced ability to diffusely invade the brain parenchyma. Various neuropeptides, including gastrin, are able to modulate tumour astrocyte migration. While neurotensin has been shown to influence the proliferation of glioma cells and the migratory ability of a large set of other cell types, its role in glioma cell migration has never been investigated. Neurotensin-induced modifications to the motility features of human U373 glioblastoma cells therefore constitute the topic of the present study. We evidenced that three subtypes of neurotensin receptors (NTR1, NTR2 and NTR3) are expressed in U373 glioblastoma cells, at least as far as their mRNAs are concerned. Treating U373 tumour cells with 10 nM neurotensin markedly modified the morphological patterns of these cells and also profoundly altered the organization of their actin cytoskeletons. Pull-down assays revealed that neurotensin induced the activation in U373 cells of both Rac1 and Cdc42 but not RhoA. Scratch wound assays evidenced that neurotensin (0.1 and 10 nM) very significantly inhibited wound colonization by U373 cells cultured in the absence of serum. In addition, quantitative phase-contrast videomicroscopy analyses showed that neurotensin decreases the motility levels of U373 glioblastoma cells when these cells are cultured on plastic. In sharp contrast, neurotensin stimulates the motility of U373 cells when they are cultured on laminin, which is a pro-adhesive extracellular matrix component ubiquitously secreted by glioma cells. Our data thus strongly suggest that, in addition to gastrin, neurotensin is a neuropeptide capable of modulating tumour astrocyte migration into the brain parenchyma. PMID- 17083473 TI - Matrix metalloproteinase-19 is highly expressed in active multiple sclerosis lesions. AB - Matrix metalloproteinases (MMPs) are proteases known for their capacity to degrade extracellular matrix (ECM) components. MMPs have been implicated in several central nervous system (CNS) diseases, including multiple sclerosis (MS). Microarray analysis has demonstrated significant increased mRNA levels of MMP-19 in chronic MS lesions, suggesting a role of MMP-19 in MS pathogenesis. Therefore, in this study, we investigated the expression pattern and cellular localization of MMP-19 protein in various well-characterized MS lesion stages. In normal control patient white matter, MMP-19 was constitutively expressed by microglia throughout the brain parenchyma, suggesting a physiological role for this MMP family member. Likewise, MMP-19 was expressed by microglia in (p)reactive MS lesions, albeit more intense. In highly active demyelinating MS lesions, parenchymal and perivascular myelin-laden macrophages were strongly immunoreactive for MMP-19, whereas reactive astrocytes were occasionally immunopositive. Astrocytes in chronic inactive lesions were weakly stained for MMP-19. In vitro, MMP-19 was expressed in cultures of primary human microglia, not in astrocyte cultures. As MMP-19 is able to degrade basement membrane constituents and other ECM proteins, it is conceivable that this relatively novel MMP family member contributes to MS pathology by remodelling the ECM of the CNS, thereby influencing leucocyte infiltration, axonal regeneration and astrogliosis. PMID- 17083474 TI - The expression of nNOS, iNOS and nitrotyrosine is increased in the rat cerebral cortex in experimental hepatic encephalopathy. AB - The changes in the distribution and amount of nitric oxide (NO) synthases (nNOS and iNOS) and the appearance of nitrotyrosine (NT) in the rat cerebral cortex were investigated following portacaval anastomosis (PCA), an experimental hepatic encephalopathy (HE) model. One month after PCA, rats showed more neurones immunoreactive to nNOS than did control animals. At 6 months post PCA, the number of neurones expressing nNOS had again increased and the intensity of the immunoreactions was stronger. Immunohistochemical analysis also showed that iNOS was increasingly expressed in pyramidal-like cortical neurones and in perivascular astrocytes from 1 to 6 months post PCA. In addition, a significant increase in cerebral iNOS concentration, at both post-PCA periods, was determined by Western blotting. The iNOS induction appears to be correlated with the length of the post-PCA period. PCA also induced the expression of NT, a nitration product of peroxynitrite. NT immunoreactivity was found in pyramidal-like cortical neurones. At 6 months, NT immunoreactivity was also evident in perivascular astrocytes, which was concomitant with a significant increase in NT protein level. PCA therefore not only increases the expression of nNOS but also induces the expression of iNOS and NT in both neurones and astrocytes. Taken together, these findings indicate that the induction of iNOS in pyramidal neurones and cortical astrocytes 6 months after PCA contributes to the generation of NT, and demonstrate the clear participation of NO in the pathogenic process of HE in this model. PMID- 17083475 TI - Differential gene expression between human schwannoma and control Schwann cells. AB - The NF2 gene encodes the tumour suppressor protein merlin. The mutation of a single allele of this gene causes the autosomal dominantly inherited disease neurofibromatosis type 2 (NF2), which is characterized mainly by vestibular schwannoma carrying a second hit mutation. Complete lack of merlin is also found in spontaneous schwannomas and meningiomas. As the events leading to schwannoma development are largely unknown we investigated the differences in gene expression between schwannoma cells from NF2 patients and normal human primary Schwann cells by cDNA array analysis. We identified 41 genes whose expression levels differed by more than factor 2. Most of these clones were corroborated by real-time reverse transcription polymerase chain reaction analysis. By this method a total of seven genes with increased and seven genes with decreased mRNA levels in schwannoma compared with normal Schwann cells could be identified. Regulated clones, some of which not been described in Schwann cells earlier, included matrix metalloproteinase's, growth factors, growth factor receptors and tyrosine kinases. PMID- 17083476 TI - Upregulation of frizzled 9 in astrocytomas. AB - Wnt/frizzled (FZD) cascades play important roles in controlling cell fate, proliferation, migration, tissue architecture and organogenesis during embryonic development and in adult organisms. The potential involvement of this pathway in tumorigenesis has been established in several types of cancers. Frizzled 9 (FZD9) is expressed in brain and its aberrant expression in gastric cancer was observed. However, its association with astrocytomas remains unknown therefore we studied FZD9 expression in astrocytomas of different malignancy. In the present study, FZD9 expression in 25 astrocytomas was investigated using immunohistochemistry with specific antibodies. Further FZD9 expression in native human brain tissue and glioblastoma cell line were analysed using real-time reverse transcription polymerase chain reaction (RT-PCR). In human astrocytomas, FZD9 immunoreactivity (IR) was observed in both microvessels and neoplastic cells. The percentage of FZD9+ microvessels in relation to FZD9+ vessels was significantly higher in malignant astrocytomas than in low-grade astrocytomas and positively correlated with the astrocytoma World Health Organization (WHO) grading (r = 1, P = 0.04). Furthermore, the FZD9 IR scores positively correlated with astrocytoma WHO grading (r = 1, P = 0.04) and proliferating activity (r = 0.77, P < 0.001). Real time RT-PCR data showed that FZD9 expression in human glioblastoma was significant higher than in normal brain (P < 0.05) but FZD9 expression was only slightly induced in cobalt chloride-treated human glioblastoma T98G cells compared with untreated cells (P > 0.05). FZD9 is upregulated in astrocytomas, suggesting that FZD9 could be important in the tumorigenesis of human astrocytomas. PMID- 17083477 TI - Paroxysmal coughing, subdural and retinal bleeding: a computer modelling approach. AB - Unexplained subdural and retinal haemorrhages in an infant are commonly attributed to 'shaking', the mechanism of which is believed to be traumatic venous rupture. However, the haemorrhagic retinopathy reported as a result of Valsalva manoeuvres and the subdural bleeding that is a rare complication of pertussis together demonstrate that if a sustained rise in intrathoracic pressure is transmitted to cerebral and retinal vessels, it may result in bleeding, similar to that reported in inflicted injury. Such haemorrhages would be expected to occur whenever severe paroxysmal coughing were induced, whatever the cause. This study used a computer modelling approach to investigate feeding accidents as the trigger for bleeding. A dynamic circulatory model of a 3-month-old infant was induced to 'cough', and the response to changes in physiological variables monitored. It showed that coughing causes intracranial pressures to build up exponentially to approach a maximum, proportional to the amount of pressure the musculature of the thorax can produce, as venous return is impeded. They do not have time to become dangerous during individual coughs, as blood quickly returns after the cough is over, reestablishing normal pressures. Paroxysmal coughing, however, does not allow blood to return between coughs, with the result that very high luminal pressures may be generated, sufficient to damage veins. A history of coughing, vomiting or choking is not uncommon in otherwise normal infants with retinal and subdural bleeding. Our findings suggest that paroxysmal coughing could account for such bleeding in some cases. PMID- 17083478 TI - Degeneration of ingestion-related brainstem nuclei in spinocerebellar ataxia type 2, 3, 6 and 7. AB - Dysphagia, which can lead to nutritional deficiencies, weight loss and dehydration, represents a risk factor for aspiration pneumonia. Although clinical studies have reported the occurrence of dysphagia in patients with spinocerebellar ataxia type 2 (SCA2), type 3 (SCA3), type 6 (SCA6) and type 7 (SCA7), there are neither detailed clinical records concerning the kind of ingestive malfunctions which contribute to dysphagia nor systematic pathoanatomical studies of brainstem regions involved in the ingestive process. In the present study we performed a systematic post mortem study on thick serial tissue sections through the ingestion-related brainstem nuclei of 12 dysphagic patients who suffered from clinically diagnosed and genetically confirmed spinocerebellar ataxias assigned to the CAG-repeat or polyglutamine diseases (two SCA2, seven SCA3, one SCA6 and two SCA7 patients) and evaluated their medical records. Upon pathoanatomical examination in all of the SCA2, SCA3, SCA6 and SCA7 patients, a widespread neurodegeneration of the brainstem nuclei involved in the ingestive process was found. The clinical records revealed that all of the SCA patients were diagnosed with progressive dysphagia and showed dysfunctions detrimental to the preparatory phase of the ingestive process, as well as the lingual, pharyngeal and oesophageal phases of swallowing. The vast majority of the SCA patients suffered from aspiration pneumonia, which was the most frequent cause of death in our sample. The findings of the present study suggest (i) that dysphagia in SCA2, SCA3, SCA6 and SCA7 patients may be associated with widespread neurodegeneration of ingestion-related brainstem nuclei; (ii) that dysphagic SCA2, SCA3, SCA6 and SCA7 patients may suffer from dysfunctions detrimental to all phases of the ingestive process; and (iii) that rehabilitative swallow therapy which takes specific functional consequences of the underlying brainstem lesions into account might be helpful in preventing aspiration pneumonia, weight loss and dehydration in SCA2, SCA3, SCA6 and SCA7 patients. PMID- 17083479 TI - Differential microglial regulation in the human spinal cord under normal and pathological conditions. AB - As the primary intrinsic immune effector cells of the central nervous system, microglia are involved in virtually all pathological processes of the brain and spinal cord including inflammatory, neurodegenerative, traumatic, neoplastic and vascular diseases. Despite this important role, there is a lack of data concerning microglial distribution and protein expression in the human spinal cord. In this study, we immunohistochemically investigated 10 normal human spinal cords to establish reference data and compared these results with 15 pathological human spinal cords deriving from distinct pathologies. Each spinal cord was evaluated at eight different levels for three white and two grey matter areas for both constitutive (MHC-II, CD68, IL-16, AIF-1, LCA, CD4) and reactive (MRP-8, MRP 14) microglial antigens. Whereas previous studies revealed significant regional differences in microglial distribution and protein expression in human brain, normal spinal cord displayed a uniform expression pattern, reaching levels of up to 17% MHC-II positive cells of the total cell population. This datum formed the basis for the further evaluation of microglia expression levels in pathological spinal cords, where levels of up to 45% positive cells were observed. Our results represent important reference values for future neuropathological diagnostic and therapeutical approaches in spinal cord pathologies. PMID- 17083480 TI - Telomerase reverse transcriptase (TERT) expression and proliferation in canine brain tumours. AB - Telomerase is a ribonucleoprotein enzyme complex that synthesizes telomere DNA. It is detected in 85-90% of malignant tumours in humans, but not in most somatic cells. Because telomerase plays a critical role in cell immortality, it represents an important target for anticancer therapies. We have previously shown that the dog is a potentially useful model for evaluating telomerase-based therapeutics. In this present study we analysed 93 canine brain tumours for telomerase reverse transcriptase (TERT) expression by immunohistochemistry. TERT immunoreactivity was detected in 16 of 50 grade 1 (32%) and 29 of 43 grade 2 tumours (67.4%), demonstrating a statistically significant association with histological grade (P = 0.00012). A subset of 51 tumours was also assessed for MIB-1 expression. The MIB-1 labelling index (LI) was found to correlate significantly with tumour grade, with a mean MIB-1 LI of 1.5% for grade 1 tumours, as compared with a mean MIB-1 LI of 21.7% for grade 2 tumours (P << 0.001). The MIB-1 LI was also significantly associated with TERT expression in all brain tumours (P << 0.001). These data further support the dog as a model for the preclinical development of telomerase-based therapeutics in brain tumours. PMID- 17083481 TI - Platelet-activating factor and human meningiomas. AB - Meningiomas are common primary intracranial tumours. Platelet-activating factor (PAF) is an inflammatory and angiogenic lipid mediator involved in several types of cancer. The presence of PAF receptor (PAF-R) transcripts, the levels of PAF, the phospholipase A2 activity (PLA2, the enzymatic activity implicated in PAF formation) and the PAF acetylhydrolase activity (AHA, the PAF degrading enzyme) were investigated in 49 human meningiomas. PAF-R transcripts, PAF, PLA2 and AHA were detected in meningiomas. However, their levels did not correlate with biological parameters such as the tumour grade, the presence of associated oedema, necrosis, mitotic index as well as intensity of the neovascularization and chronic inflammatory response. In conclusion, PAF is present in meningiomas where it might act on tumour growth by altering the local angiogenic and/or cytokine networks as previously suggested for human breast and colorectal cancer. PMID- 17083482 TI - Commentary on: Return of the cycad hypothesis--does the amyotrophic lateral sclerosis/Parkinsonism dementia complex (ALS/PDC) of Guam have new implications for global health? PMID- 17083483 TI - Editorial: beyond the pale. PMID- 17083484 TI - Hypopigmenting agents: an updated review on biological, chemical and clinical aspects. AB - An overview of agents causing hypopigmentation in human skin is presented. The review is organized to put forward groups of biological and chemical agents. Their mechanisms of action cover (i) tyrosinase inhibition, maturation and enhancement of its degradation; (ii) Mitf inhibition; (iii) downregulation of MC1R activity; (iv) interference with melanosome maturation and transfer; (v) melanocyte loss, desquamation and chemical peeling. Tyrosinase inhibition is the most common approach to achieve skin hypopigmentation as this enzyme catalyses the rate-limiting step of pigmentation. Despite the large number of tyrosinase inhibitors in vitro, only a few are able to induce effects in clinical trials. The gap between in-vitro and in-vivo studies suggests that innovative strategies are needed for validating their efficacy and safety. Successful treatments need the combination of two or more agents acting on different mechanisms to achieve a synergistic effect. In addition to tyrosinase inhibition, other parameters related to cytotoxicity, solubility, cutaneous absorption, penetration and stability of the agents should be considered. The screening test system is also very important as keratinocytes play an active role in modulating melanogenesis within melanocytes. Mammalian skin or at least keratinocytes/melanocytes co cultures should be preferred rather than pure melanocyte cultures or soluble tyrosinase. PMID- 17083485 TI - The physical and chemical properties of eumelanin. AB - In this article, we review the current state of knowledge concerning the physical and chemical properties of the eumelanin pigment. We examine properties related to its photoprotective functionality, and draw the crucial link between fundamental molecular structure and observable macroscopic behaviour. Where necessary, we also briefly review certain aspects of the pheomelanin literature to draw relevant comparison. A full understanding of melanin function, and indeed its role in retarding or promoting the disease state, can only be obtained through a full mapping of key structure-property relationships in the main pigment types. We are engaged in such an endeavor for the case of eumelanin. PMID- 17083486 TI - Activation of the Mitf promoter by lipid-stimulated activation of p38-stress signalling to CREB. AB - The microphthalmia-associated transcription factor Mitf plays a critical role in regulating many aspects of melanocyte biology. It is required for melanoblast and postnatal melanocyte survival, regulates proliferation, and activates genes associated with differentiation such as tyrosinase and related genes involved in melanogenesis. Identifying the signals that regulate Mitf expression is crucial if we are to understand how cells of the melanocyte lineage respond to environmental cues. Here we show that the Mitf promoter is induced by lipid signalling via the p38 stress-activated kinase pathway that is also activated by a wide range of receptors as well as UV irradiation. Signalling via p38 leads to increased phosphorylation and activation of cyclic adenosine monophosphate response element-binding (CREB) that binds and activates the Mitf promoter via the cyclic adenosine monophosphate (cAMP) response element. Moreover, we also show that activation of p38 mediated by lipids is potentiated by inhibition of the PI3kinase pathway but not by inhibition of protein kinase A (PKA). The results identify a mechanism in which stress signalling via p38 leads to activation of CREB, enhanced Mitf expression and consequently increased tyrosinase expression. The results are relevant for the regulation of melanocytes by Mitf, but also raise the possibility that lipid mediated activation of p38 signalling may represent a potential therapy for vitiligo. PMID- 17083487 TI - Relationship between skin response to ultraviolet exposure and skin color type. AB - Sun exposure is responsible for detrimental damage ranging from sunburn to photoaging and skin cancer. This damage is likely to be influenced by constitutive pigmentation. The relationship between ultraviolet (UV) sensitivity and skin color type was analyzed on 42 ex vivo skin samples objectively classified from light to dark skin, based on their values of individual typology angle (ITA) determined by colorimetric parameters. The biologically efficient dose (BED) was determined for each sample by quantifying sunburn cells after exposure to increasing doses of UV solar-simulated radiation. Typical UV-induced biologic markers, other than erythema, such as DNA damage, apoptosis and p53 accumulation, were analyzed. A statistically significant correlation was found between ITA and BED and, ITA and DNA damage. Interestingly, DNA lesions were distributed throughout the whole epidermal layers and the uppermost dermal cells in light, intermediate and tanned skin while they were restricted to suprabasal epidermal layers in brown or dark skin. Our data support, at the cellular level, the relationship between UV sensitivity and skin color type. They emphasize the impact of DNA damage accumulation in basal layer in relation to the prevalence of skin cancer. PMID- 17083488 TI - The Fugu tyrp1 promoter directs specific GFP expression in zebrafish: tools to study the RPE and the neural crest-derived melanophores. AB - In vertebrates, pigment cells account for a small percentage of the total cell population and they intermingle with other cell types. This makes it difficult to isolate them for analyzes of their functions in the context of development. To alleviate such difficulty, we generated two stable transgenic zebrafish lines (pt101 and pt102) that express green fluorescent protein (GFP) in melanophores under the control of the 1 kb Fugu tyrp1 promoter. In pt101, GFP is expressed in both retinal pigment epithelium (RPE) cells and the neural crest-derived melanophores (NCDM), whereas in pt102, GFP is predominately expressed in the NCDM. Our results indicate that the Fugu tyrp1 promoter can direct transgene expression in a cell-type-specific manner in zebrafish. In addition, our findings provide evidence supporting differential regulations of melanin-synthesizing genes in RPE cells and the NCDM in zebrafish. Utilizing the varying GFP expression levels in these fish, we have isolated melanophores via flow cytometry and revealed the capability of sorting the NCDM from RPE cells as well. Thus, these transgenic lines are useful tools to study melanophores in zebrafish. PMID- 17083489 TI - Reduced expression of vps11 causes less pigmentation in medaka, Oryzias latipes. AB - In this article, we describe the medaka mutant pale gray eyes (pge) that shows reduction of black, white, and silver pigmentation and lethality approximately a week after hatching. The pge mutation was mapped to the tip of linkage group 14 and no recombinations were observed between the mutation and medaka vps11 in 900 meioses. Vps 11 is one of the evolutionarily conserved class C vacuolar protein sorting genes (c-vps: vps11, vps16, vps18, and vps33), whose products physically associate to form the c-vps protein complex required for vesicle docking and fusion in the budding yeast. Mutations in vps16, vps18, and vps33 are known to result in decreased pigmentation in organisms such as Drosophila. We cloned the full-length medaka vps11 cDNA by rapid amplification of cDNA ends (RACE) and found no RACE products from the pge mutants. Similarly, no vps11 transcripts were detected from the pge mutants by Northern analysis. The injection of an antisense morpholino against vps11 phenocopied the pge mutant. Taken together, the results suggest that reduced expression of medaka vps11 causes pge and that medaka vps11 is indispensable for survival and normal pigmentation. PMID- 17083490 TI - Characterization of Mahogunin Ring Finger-1 expression in mice. AB - Mutations in mouse Mahogunin Ring Finger-1 (Mgrn1) were first recognized for their effect on agouti-mediated pigment-type switching. Mgrn1 null mutants are completely black and develop spongiform degeneration of the brain. Mgrn1 hypomorphs have dark fur but do not develop neurodegeneration. We characterized a new Mgrn1 hypomorphic allele caused by a gene-trap insertion. Mice homozygous for this mutation are slightly darker than non-mutant animals. They show reduced overall expression of Mgrn1 and two of the four normal Mgrn1 isoforms are replaced by beta-GEO fusion proteins that differ from the normal proteins at their carboxy termini. To investigate the role of different Mgrn1 isoforms in pigment-type switching, we used quantitative relative reverse transcriptase polymerase chain reaction to examine their expression in the skin of Mgrn1 mutant and control mice. Most Mgrn1 mutants produce little or no normal Mgrn1 in the skin. Mgrn1 null mutant mice overexpressing isoform I or III, which are normally absent or weakly expressed in adult skin, had normal agouti-banded hairs. Our results indicate that reduced levels of MGRN1 cause the pigmentation phenotypes of Mgrn1 mutant mice and that there are no significant differences in the function of the four MGRN1 isoforms in pigment-type switching. PMID- 17083491 TI - Genomic localization of a Dct-LacZ transgene locus: a simple assay for transgene status. PMID- 17083493 TI - Abstracts from the 2006 Annual Meeting of the American Epilepsy Society and the Canadian League against Epilepsy (1st North American Regional Epilepsy Congress). PMID- 17083495 TI - Changes in religiousness and spirituality attributed to HIV/AIDS: are there sex and race differences? AB - BACKGROUND/OBJECTIVE: Having a serious illness such as HIV/AIDS raises existential issues, which are potentially manifested as changes in religiousness and spirituality. The objective of this study was (1) to describe changes in religiousness and spirituality of people with HIV/AIDS, and (2) to determine if these changes differed by sex and race. METHODS: Three-hundred and forty-seven adults with HIV/AIDS from 4 sites were asked demographic, clinical, and religious/spiritual questions. Six religious/spiritual questions assessed personal and social domains of religiousness and spirituality. RESULTS: Eighty eight participants (25%) reported being "more religious" and 142 (41%) reported being "more spiritual" since being diagnosed with HIV/AIDS. Approximately 1 in 4 participants also reported that they felt more alienated by a religious group since their HIV/AIDS diagnosis and approximately 1 in 10 reported changing their place of religious worship because of HIV/AIDS. A total of 174 participants (50%) believed that their religiousness/spirituality helped them live longer. Fewer Caucasians than African Americans reported becoming more spiritual since their HIV/AIDS diagnosis (37% vs 52%, respectively; P<.015), more Caucasians than African Americans felt alienated from religious communities (44% vs 21%, respectively; P<.001), and fewer Caucasians than African Americans believed that their religiousness/spirituality helped them live longer (41% vs 68% respectively; P<.001). There were no significantly different reported changes in religious and spiritual experiences by sex. CONCLUSIONS: Many participants report having become more spiritual or religious since contracting HIV/AIDS, though many have felt alienated by a religious group-some to the point of changing their place of worship. Clinicians conducting spiritual assessments should be aware that changes in religious and spiritual experiences attributed to HIV/AIDS might differ between Caucasian and African Americans. PMID- 17083496 TI - Religion, spirituality, and depressive symptoms in patients with HIV/AIDS. AB - BACKGROUND: Depression has been linked to immune function and mortality in patients with chronic illnesses. Factors such as poorer spiritual well-being has been linked to increased risk for depression and other mood disorders in patients with HIV. OBJECTIVE: We sought to determine how specific dimensions of religion, spirituality, and other factors relate to depressive symptoms in a contemporary, multi-center cohort of patients with HIV/AIDS. DESIGN: Patients were recruited from 4 medical centers in 3 cities in 2002 to 2003, and trained interviewers administered the questionnaires. The level of depressive symptoms was measured with the 10-item Center for Epidemiologic Studies Depression (CESD-10) Scale. Independent variables included socio-demographics, clinical information, 8 dimensions of health status and concerns, symptoms, social support, risk attitudes, self-esteem, spirituality, religious affiliation, religiosity, and religious coping. We examined the bivariate and multivariable associations of religiosity, spirituality, and depressive symptoms. MEASUREMENTS AND MAIN RESULTS: We collected data from 450 subjects. Their mean (SD) age was 43.8 (8.4) years; 387 (86.0%) were male; 204 (45.3%) were white; and their mean CD4 count was 420.5 (301.0). Two hundred forty-one (53.6%) fit the criteria for significant depressive symptoms (CESD-10 score > or = 10). In multivariable analyses, having greater health worries, less comfort with how one contracted HIV, more HIV related symptoms, less social support, and lower spiritual well-being was associated with significant depressive symptoms (P<.05). CONCLUSION: A majority of patients with HIV reported having significant depressive symptoms. Poorer health status and perceptions, less social support, and lower spiritual well being were related to significant depressive symptoms, while personal religiosity and having a religious affiliation was not associated when controlling for other factors. Helping to address the spiritual needs of patients in the medical or community setting may be one way to decrease depressive symptoms in patients with HIV/AIDS. PMID- 17083497 TI - Modeling the effects of spirituality/religion on patients' perceptions of living with HIV/AIDS. AB - BACKGROUND: Spirituality/religion is an important factor in health and illness, but more work is needed to determine its link to quality of life in patients with HIV/AIDS. OBJECTIVE: To estimate the direct and indirect effects of spirituality/religion on patients' perceptions of living with HIV/AIDS. DESIGN: In 2002 and 2003, as part of a multicenter longitudinal study of patients with HIV/AIDS, we collected extensive demographic, clinical, and behavioral data from chart review and patient interviews. We used logistic regression and path analysis combining logistic and ordinary least squares regression. SUBJECTS: Four hundred and fifty outpatients with HIV/AIDS from 4 sites in 3 cities. MEASURES: The dependent variable was whether patients felt that life had improved since being diagnosed with HIV/AIDS. Spirituality/religion was assessed by using the Duke Religion Index, Functional Assessment of Chronic Illness Therapy-Spiritual Well-Being-Expanded, and Brief RCOPE measures. Mediating factors included social support, self-esteem, healthy beliefs, and health status/health concerns. RESULTS: Approximately one-third of the patients felt that their life was better now than it was before being diagnosed with HIV/AIDS. A 1-SD increase in spirituality/religion was associated with a 68.50% increase in odds of feeling that life has improved--29.97% due to a direct effect, and 38.54% due to indirect effects through healthy beliefs (29.15%) and health status/health concerns (9.39%). Healthy beliefs had the largest effect on feeling that life had improved; a 1-SD increase in healthy beliefs resulted in a 109.75% improvement in feeling that life changed. CONCLUSIONS: In patients with HIV/AIDS, the level of spirituality/religion is associated, both directly and indirectly, with feeling that life is better now than previously. Future research should validate our new conceptual model using other samples and longitudinal studies. Clinical education interventions should focus on raising awareness among clinicians about the importance of spirituality/religion in HIV/AIDS. PMID- 17083498 TI - The importance of spirituality/religion and health-related quality of life among individuals with HIV/AIDS. PMID- 17083499 TI - Health-related quality of life in veterans and nonveterans with HIV/AIDS. AB - PURPOSE: To compare health-related quality of life (HRQoL) between patients receiving care in Veterans Administration (VA) settings (veterans) and non-VA settings (nonveterans), and to explore determinants of HRQoL and change in HRQoL over time in subjects living with HIV/AIDS. SUBJECTS: One hundred veterans and 350 nonveterans with HIV/AIDS from 2 VA and 2 university-based sites in 3 cities interviewed in 2002 to 2003 and again 12 to 18 months later. METHODS: We assessed health status (functional status and symptom bother), health ratings, and health values (time tradeoff [TTO] and standard gamble [SG] utilities). We also explored bivariate and multivariable associations of HRQoL measures with a number of demographic, clinical, spiritual/religious, and psychosocial characteristics. RESULTS: Compared with nonveterans, the veteran population was older (47.7 vs 42.0 years) and consisted of a higher proportion of males (97% vs 83%), of participants with a history of injection drug use (23% vs 15%), and of subjects with unstable housing situations (14% vs 6%; P<.05 for all comparisons). On scales ranging from 0 (worst) to 100 (best), veterans reported significantly poorer overall function (mean [SD]; 65.9 [17.2] vs 71.9 [16.8]); lower rating scale scores (67.6 [21.7] vs 73.5 [21.0]), lower TTO values (75.7 [37.4] vs 89.0 [23.2]), and lower SG values (75.0 [35.8] vs 83.2 [28.3]) than nonveterans (P<.05 for all comparisons); however, in multivariable models, veteran status was only a significant determinant of SG and TTO values at baseline. Among other determinants that were associated with multiple HRQoL outcomes in baseline and follow-up multivariable analyses were: symptom bother, overall function, religiosity/spirituality, depressive symptoms, and financial worries. CONCLUSIONS: Veterans reported significantly poorer HRQoL than nonveterans, but when controlling for other factors, veteran status was only a significant determinant of TTO and SG health values at baseline. Correlates of HRQoL such as symptom bother, spirituality/religiosity, and depressive symptoms could be fruitful potential targets for interventions to improve HRQoL in patients with HIV/AIDS. PMID- 17083500 TI - Patterns of responses on health-related quality of life questionnaires among patients with HIV/AIDS. AB - BACKGROUND: Health-related quality of life (HRQoL) has become an important facet of HIV/AIDS research. Typically, the unit of analysis is either the total instrument score or subscale score. Developing a typology of responses across various HRQoL measures, however, may advance understating of patients' perspectives. METHODS: In a multicenter study, we categorized 443 patients' responses on utility measures (time-tradeoff, standard gamble, and rating scale) and the HIV/AIDS-Targeted Quality of Life (HAT-QoL) scale by using latent profile analysis to empirically derive classes of respondents. We then used linear regressions to identify whether class membership is associated with clinical measures (viral load, CD4, time since diagnosis, highly active antiretroviral therapy [HAART]) and psychosocial function (depressed mood, alcohol use, religious coping). RESULTS: Six classes were identified. Responses across the HAT QoL subscales tended to fall into 3 groupings--high functioning (Class 1), moderate functioning (Classes 2 and 3), and low functioning (Classes 4 to 6); utility measures further distinguished individuals among classes. Regression analyses comparing those in Class 1 with those in the other 5 found significantly more symptoms of depression, negative religious coping strategies, and lower CD4 counts among subjects in Class 1. Those in Class 5 had been diagnosed with HIV longer, and members of Class 6 reported significantly less alcohol consumption, had higher viral loads, and were more likely to receive HAART. CONCLUSION: Patients with HIV respond differentially to various types of HRQoL measures. Health status and utility measures are thus complementary approaches to measuring HRQoL in patients with HIV. PMID- 17083501 TI - Spirituality and religion in patients with HIV/AIDS. AB - BACKGROUND: Spirituality and religion are often central issues for patients dealing with chronic illness. The purpose of this study is to characterize spirituality/religion in a large and diverse sample of patients with HIV/AIDS by using several measures of spirituality/religion, to examine associations between spirituality/religion and a number of demographic, clinical, and psychosocial variables, and to assess changes in levels of spirituality over 12 to 18 months. METHODS: We interviewed 450 patients from 4 clinical sites. Spirituality/religion was assessed by using 8 measures: the Functional Assessment of Chronic Illness Therapy-Spirituality-Expanded scale (meaning/peace, faith, and overall spirituality); the Duke Religion Index (organized and nonorganized religious activities, and intrinsic religiosity); and the Brief RCOPE scale (positive and negative religious coping). Covariates included demographics and clinical characteristics, HIV symptoms, health status, social support, self-esteem, optimism, and depressive symptoms. RESULTS: The patients' mean (SD) age was 43.3 (8.4) years; 387 (86%) were male; 246 (55%) were minorities; and 358 (80%) indicated a specific religious preference. Ninety-five (23%) participants attended religious services weekly, and 143 (32%) engaged in prayer or meditation at least daily. Three hundred thirty-nine (75%) patients said that their illness had strengthened their faith at least a little, and patients used positive religious coping strategies (e.g., sought God's love and care) more often than negative ones (e.g., wondered whether God has abandoned me; P<.0001). In 8 multivariable models, factors associated with most facets of spirituality/religion included ethnic and racial minority status, greater optimism, less alcohol use, having a religion, greater self-esteem, greater life satisfaction, and lower overall functioning (R2=.16 to .74). Mean levels of spirituality did not change significantly over 12 to 18 months. CONCLUSIONS: Most patients with HIV/AIDS belonged to an organized religion and use their religion to cope with their illness. Patients with greater optimism, greater self-esteem, greater life satisfaction, minorities, and patients who drink less alcohol tend to be both more spiritual and religious. Spirituality levels remain stable over 12 to 18 months. PMID- 17083502 TI - How do patients with HIV/AIDS understand and respond to health value questions? AB - BACKGROUND: Utility assessment involves assigning values to experienced or unfamiliar health states. Pivotal to utility assessment, then, is how one conceptualizes health states such as "current health" and "perfect health." The purpose of this study was to ascertain how patients with HIV think about and value health and health states. METHODS: We conducted open-ended in-depth interviews with 32 patients with HIV infection purposefully sampled from a multicenter study of quality of life in HIV. After undergoing computer-assisted utility assessment using the rating scale, time tradeoff, and standard gamble methods, patients were asked how they thought about the utility tasks and about the terms "current health" and "perfect health." RESULTS: Patients understood the health valuation tasks but conceptualized health states in different ways. Many patients believed that "perfect health" was a mythical health state, and some questioned whether it was even desirable. "Current health" was variably interpreted as the status quo; deteriorating over time; or potentially improving with the hope of a cure. CONCLUSION: Patients with HIV infection vary in the way they conceptualize health states central to utility assessment, such as perfect health and current health. Better understanding of these issues could make important methodologic and policy-level contributions. PMID- 17083503 TI - An increase in religiousness/spirituality occurs after HIV diagnosis and predicts slower disease progression over 4 years in people with HIV. AB - BACKGROUND: Most studies on religion/spirituality predicting health outcomes have been limited to church attendance as a predictor and have focused on healthy people. However, confronting a major medical crisis may be a time when people turn to the sacred. OBJECTIVE: The purpose of this study was to determine the extent to which changes in spirituality/religiousness occur after HIV diagnosis and whether changes predict disease progression. DESIGN/PARTICIPANTS: This longitudinal study examined the relationship between changes in spirituality/religiousness from before with after the diagnosis of HIV, and disease progression (CD4 and viral load [VL] every 6 months) over 4 years in 100 people with HIV. Measures included change in religiousness/spirituality after diagnosis of HIV, religiousness/spirituality at various times in one's life, church attendance, depression, hopelessness, optimism, coping (avoidant, proactive), social support, CD4/VL, and health behaviors. RESULTS: Forty-five percent of the sample showed an increase in religiousness/spirituality after the diagnosis of HIV, 42% remained the same, and 13% decreased. People reporting an increase in spirituality/religiousness after the diagnosis had significantly greater preservation of CD4 cells over the 4-year period, as well as significantly better control of VL. Results were independent of (i.e., held even after controlling for) church attendance and initial disease status (CD4/VL), medication at every time point, age, gender, race, education, health behaviors (adherence, risky sex, alcohol, cocaine), depression, hopelessness, optimism, coping (avoidant, proactive), and social support. CONCLUSIONS: There is an increase in spirituality/religiousness after HIV diagnosis, and this increase predicts slower disease progression; medical personnel should be aware of its potential importance. PMID- 17083504 TI - Commentary: 'switching between full-length and B-domain deleted factor VIII and the risk of inhibitors'. PMID- 17083505 TI - The 80th anniversary of von Willebrand's disease: history, management and research. AB - The history of von Willebrand's disease (VWD) is fascinating because it demonstrates how good clinical observations, genetic studies and biochemical skills can improve basic understanding of a disease and its management. The continuous efforts of scientists and clinicians during the last 80 years have significantly improved the knowledge of von Willebrand factor (VWF) structure and function and the management of VWD. Diagnosis of phenotype and genotype is now available in many countries and treatment is becoming more specific according to the VWD type. Any therapeutic agents must correct the dual defect of haemostasis, i.e. the abnormal platelet adhesion due to reduced and/or dysfunctional and low levels of factor VIII (FVIII) associated with VWF defects. Desmopressin (DDAVP) is the treatment of choice for type 1 VWD because it induces release of VWF from cellular compartments. Plasma virally inactivated VWF concentrates containing FVIII are effective and safe in patients unresponsive to DDAVP. There are advanced plans to develop a recombinant VWF but this product will require the concomitant administration of FVIII for the control of acute bleeds. Basic research studies on cellular biology, biochemistry and immunology have confirmed the role of VWF as a crucial participant in both haemostasis and thrombosis as its main biological activity is to support platelet adhesion-aggregation in the circulation. Retrospective and prospective clinical research studies, including bleeding history and laboratory markers for diagnosis as well as the use of DDAVP and VWF concentrates to manage or prevent bleeds in patients with VWD have been essential to provide general guidelines for VWD management. The large number of publications quoting VWD and VWF emphasizes the important role of VWF in medicine. PMID- 17083506 TI - The need for previously untreated patient population studies in understanding the development of factor VIII inhibitors. AB - The presence of inhibitory antibodies to factor VIII (FVIII) remains one of the most serious complications of haemophilia therapy. Accordingly, understanding risk factors that may contribute to inhibitor developments in young patients with haemophilia A continues to be an area of great interest. Previously untreated patient (PUP) population studies have been instrumental in understanding the aetiology of inhibitor development. These studies have revealed the importance of risk factors such as clotting factor exposure history, ethnicity, and FVIII genotype in the development of inhibitors, while also providing insights into potential risk factors that may be related to therapeutic practice. However, due to differences in study designs and patient populations among previous PUP studies, there are limitations to the value of these studies in deciphering the role of potential risk factors. Therefore, future PUP studies should be prospective, consistent in their study designs and consider all established parameters and also those that possibly may influence inhibitor formation, thereby facilitating a better understanding of the aetiology of inhibitor formation in haemophilia A patients. PMID- 17083507 TI - Epidemiology of inhibitor formation with recombinant factor VIII replacement therapy. AB - This article reviews the epidemiology of inhibitor formation in patients receiving recombinant factor VIII (rFVIII) replacement therapy for haemophilia A. Data from pivotal trial programmes, post-marketing surveillance studies, and pharmacovigilance studies for all available rFVIII products were reviewed. To date, no comparative studies have been performed, and in the non-comparative studies, there were differences in the patient population enrolled and study design (including the number of patient exposure days). In the absence of comparative clinical trials, it is not possible to make comparisons between inhibitor data for the various rFVIII products. This review of the epidemiological data shows that across the pivotal trial programmes for rFVIII products, the observed incidence of inhibitors was in the range of 15-32% in previously untreated patients (PUPs) and 0.9-2.9% in previously treated patients (PTPs). High-titre inhibitors (peak >5 BU) were detected in 10-16% of PUPs and 0 2.3% of PTPs. Several initiatives proposed to help standardize collection and interpretation of inhibitor data for patients receiving rFVIII treatment are described. Such standardization would help to clarify the epidemiology of inhibitor formation across FVIII treatments. PMID- 17083508 TI - Application of current knowledge to the management of bleeding events during immune tolerance induction. AB - The development of inhibitors to factor VIII is the most serious adverse event associated with the treatment of haemophilia A, predisposing patients to uncontrollable haemorrhage, disability and premature death. Eradication of inhibitors via immune tolerance induction (ITI) is effective in the majority of patients, but may require months to years to achieve success. In the interim, the treatment and prevention of acute bleeding episodes are primary foci of care. Regrettably, there is a paucity of information regarding management of bleeding episodes in inhibitor patients undergoing tolerization. Until specific data from ongoing clinical trials are available to provide more guidance in this patient group, it is reasonable and useful to rely on the broader base of medical literature pertaining to patients not being tolerized to deduce strategies for controlling acute and perioperative bleeding episodes in inhibitor patients during ITI. PMID- 17083509 TI - Individualization of bypassing agent treatment for haemophilic patients with inhibitors utilizing thromboelastography. AB - The treatment of bleeding for haemophilic patients with inhibitors relies on the use of the bypassing agents, recombinant factor VIIa and factor eight inhibitor bypass activity (FEIBA). While both therapies are effective in the majority of bleeding episodes, there is a significant amount of interindividual variability when it comes to the response to therapy. As of yet, there is no reliable laboratory parameter that can predict the response to therapy in the same manner that factor VIII and factor IX levels predict response in non-inhibitor patients. Developing such a laboratory parameter is vital in order to maximize the clinical efficacy of these agents. Thromboelastography (TEG) is a device, which assesses clot formation over time in whole blood and has several characteristics which suggest it may be an effective way to monitor bypass agent therapy. We studied the ability of TEG to individualize the treatment regimens of three patients with high titre inhibitors assessing the response to recombinant factor VIIa, FEIBA, and when both were used sequentially. The TEG allowed for individualization of treatment for each of the three patients and resulted in more effective, convenient and less expensive treatment regimens. We thus believe that TEG is a promising device for monitoring of bypass agent therapy and should be studied further. PMID- 17083510 TI - Influence of factor VIII on overall coagulability and fibrinolytic potential of haemophilic plasma as measured by global assay: monitoring in haemophilia A. AB - The objectives of the present study were to evaluate the analytical sensitivity of the recently developed Clot Formation and Lysis (CloFAL) global assay for factor VIII (FVIII) deficiency, both in vitro and ex vivo, to determine whether this global assay is influenced by FVIII inhibitors, and to investigate the coagulative response to FVIII replacement in haemophilia A patients using the CloFAL assay in comparison with FVIII activity. Among adults and children alike, the CloFAL assay coagulation index (CI) was significantly decreased in FVIII deficient vs. healthy subjects (adults median CI: 2% vs. 94% respectively; children median CI: 3% vs. 63%; P < 0.001 for each), and correlated significantly with activated partial thromboplastin time-based FVIII activity across all individuals (r = 0.78; P < 0.001). The CloFAL assay was analytically sensitive to deficient FVIII activity and also influenced by the presence of von Willebrand factor. Severe haemophilia A patients without inhibitory antibodies to FVIII showed considerable heterogeneity in CloFAL assay waveforms, despite a uniformly diminished CI of 0-1%. During FVIII infusion half-life studies in patients with severe haemophilia A, the CloFAL assay demonstrated a marked rise in coagulability 30 min following infusion, with progressive decrease in coagulability towards baseline over the ensuing 48-h period. In each case, the profile of coagulative response to FVIII infusion as determined by CloFAL assay CI differed qualitatively from that measured by FVIII activity. These findings indicate that the CloFAL assay may be useful as an adjunctive test to FVIII activity measurements in the therapeutic monitoring of haemophilia A. PMID- 17083511 TI - Identification of three FGA mutations in two Chinese families with congenital afibrinogenaemia. AB - Congenital afibrinogenaemia is a rare autosomal recessive disorder, characterized by the complete absence or extremely reduced level of fibrinogen (Fg). We attempted to analyse the phenotype and genotype in two Chinese families with congenital afibrinogenaemia. Coagulation studies including activated partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT) and Fg were performed in the patients and other family members. All the exons, exon intron boundaries and promoter regions of three Fg genes (FGA, FGB and FGG) were screened by direct sequencing. Three patients in two families suffered from moderate to severe haemorrhage. Their APTT, PT and TT were extremely prolonged and plasma Fg levels were undetectable by Clauss method and extremely reduced by immunoassay. Genetic analysis revealed three FGA mutations in three patients including one novel mutation. In family 1, patient 1 was detected compound heterozygous mutations in FGA, g.1892-1899delAGTA/GTAA from her patriline and g.1978-g.3215del1238 bp from her matriline. In family 2, a homozygous Gln203X in Aalpha-chain was found in both patients 2 and 3 due to consanguineous marriage. All these mutations were null mutations, which could produce premature stop codons in FGA. It can be indicated that with more genetic analysis performed on afibrinogenaemia patients all over the world, there is no distinct difference in geographical distribution of Fg gene mutations. Gln203X in Aalpha-chain was first reported in this study, which may help to further understand the function of Aalpha-chain. PMID- 17083512 TI - Severe factor X deficiency due to a homozygous mutation (Cys364Arg) that disrupts a disulphide bond in the catalytic domain. AB - Severe factor X deficiency (<0.01 IU mL(-1)) is a rare disorder producing a major bleeding tendency including umbilical cord, joint and intracranial haemorrhage. We present the first case of a child homozygous for a g.1177T > C missense alteration, predicted to disrupt the catalytic domain, and resulting in severe FX deficiency. The child suffered intracranial haemorrhage and now receives regular prophylaxis with a prothrombin complex concentrate. Our experience and a review of the literature suggest that optimal frequency of dosing is likely to be two or three times weekly and that the risk of thrombosis is very small. PMID- 17083513 TI - Screening for factor XI deficiency amongst pregnant women of Ashkenazi Jewish origin. AB - A pilot study was conducted over a 6-month period to evaluate antenatal screening for factor XI (FXI) deficiency amongst Ashkenazi Jewish women booking for their pregnancy in a single obstetric unit. Fifty-four women of Ashkenazi Jewish origin were recruited during their visit for the routine first trimester ultrasound scan. They completed a questionnaire about their personal bleeding symptoms and had blood taken for FXI levels (FXI:C). Seven (13%) women had partial FXI deficiency. Five (9%) were newly diagnosed, and in the remaining two, the diagnosis was known previously. One infant with severe FXI deficiency was identified as a result of maternal testing. This study has shown that FXI deficiency is common amongst women of Ashkenazi Jewish origin and supports its antenatal screening in this population. However, further studies are required to evaluate its cost-effectiveness and the effect on pregnancy outcome. PMID- 17083514 TI - Surface electrical stimulation of the quadriceps femoris in patients affected by haemophilia A. AB - Eighteen sessions of surface electrical stimulation was applied to the quadriceps femoris of the left leg of ten male subjects affected by severe haemophilia A, while ten healthy subjects constituted the control group. The isometric strength, the electromyographic activity and the diameter of the rectus femoris were measured in both legs before and after a six-week treatment period. After the treatment, the people affected by haemophilia showed a gain in strength by 13.8% in the stimulated leg and by 17.1% in the non-stimulated one. No changes were detected in the electromyographic activity. On the contrary, the diameter of the rectus femoris of the stimulated leg increased in 24.34%, while no significant change was found in the nonelectrically stimulated leg. These results show for the first time that the application of electrical stimulation in haemophilic patients contributes to the gain and development of strength and trophism. The results also show that the surface electrical stimulation does not represent a threat to the patients' health, and that can be used for therapeutic purposes. PMID- 17083515 TI - Health-related quality of life in chronic coagulation disorders. AB - Measurement of health-related quality of life (HR-QoL) is used in patients with haemophilia as a way of assessing the effectiveness of health care, especially as cure is not possible. We report the first such study on patients with chronic coagulation disorders in Australia, using The RAND 36-item Health Survey 1.0 (SF 36), a standardized validated questionnaire combined with a semistructured interview. The mean scores for the eight domains of the SF-36 ranged from 52.5 +/ 42.1 for physical role to 80.0 +/- 20.0 for social functioning. Comparison with normative data obtained from the Australian Bureau of Statistics (ABS) demonstrated a reduction in all domains in this population with statistically significant reductions in general health, physical role limitation and vitality. Comparison with other studies indicates that the HR-QoL of patients with haemophilia and von Willebrand's disorder in Australia is comparable with other Western countries, reflecting the overall similar quality of care available to these patients. This study also provides a good cross-sectional view of the psychosocial factors of life in patients in Australia and recognizes the positive family support available to this population, while raising relevant shortcomings in schools and at the workplace that need to be studied further in a controlled manner. PMID- 17083516 TI - Comparing two measures of quality of life for children with haemophilia: the CHO KLAT and the Haemo-QoL. AB - Disease-specific measures of quality of life (QoL) for children with haemophilia are now available for use in clinical studies [Haemophilia, 10, 2004, 9-16]. One of these measures, the Canadian Haemophilia Outcomes - Kids' Life Assessment Tool (CHO-KLAT), was developed in Canada with emphasis on the perspectives of children [Pediatr Blood Cancer, 47, 2006, 305-11; Haemophilia, 10, 2004, 34-43]. Another, the Haemo-QoL, was developed in Europe, with emphasis on the perspectives of clinicians [Haemophilia, 8, 2002, 47-54; Haemophilia, 10, 2004, 17-25]. While these two measures are unique and independent, researchers from both studies were collaboratively linked throughout development and testing. This study presents the results of a joint assessment of the two measures with respect to their strengths, limitations and unique contributions. The primary questions addressed were: 1 What is the relationship between the CHO-KLAT and the Haemo-QoL in terms of summary scores and item content? 2 What are the methodological strengths, limitations and unique contributions of each measure? We conducted a retrospective analysis of data from field testing of both measures. The analysis included a comparative assessment of the basic validity, reliability and items used in each measure. Overall, the CHO-KLAT and the Haemo-QoL are promising and valuable measures of QoL for children with haemophilia. Our analyses confirmed the basic psychometric properties of both tools, but identified some discrepancies between them. Additional data will allow for greater understanding of these discrepancies and lend clarity to how the tools should be used in clinical studies (separately or merged). The present recommendation is that the measures be run independently, but preferably concurrently in studies of children with haemophilia. PMID- 17083517 TI - Histological changes in murine haemophilic synovitis: a quantitative grading system to assess blood-induced synovitis. AB - Haemophilia is a congenital disorder that results in frequent bleeding into joints, in which a chronic and debilitating arthritis develops. The presence of blood evokes an inflammatory and proliferative synovial reaction. Although the molecular mechanisms and biochemical pathways which underlie this disorder are not known, significant advances have been made by studying a murine model of human haemophilic synovitis. In order to better understand and correlate the pathological, molecular and biochemical changes, it has become necessary to grade the histological changes observed. Despite a search of the literature and review of relevant publications, none of the currently utilized schemes were appropriate, and therefore a novel grading scheme was developed. After review of over 1000 histological sections, six characteristic changes were identified: (i) synovial hyperplasia; (ii) vascularity; (iii) discolouration by haemosiderin; (iv) the presence of blood (erythrocytes); (v) villus formation; and (vi) cartilage erosion. Synovial hyperplasia and vascularity were present in variable amounts and were quantitatively scored (0-3), while the other changes were qualitatively scored as absent or present (0 or 1). Application of the grading scheme was tested and a high interobserver correlation (greater than 80%) was found. The scheme was easy to learn even by novices, with no prior experience. The availability of the histological grading scheme for murine synovitis will allow for precise evaluation of the pathological changes following joint bleeding, and facilitate correlations with molecular and biochemical changes that lead to these changes. PMID- 17083518 TI - Rituximab treatment of mild haemophilia A with inhibitors: a proposed treatment protocol. AB - Inhibitors are an uncommon complication of mild haemophilia A but represent a severe disease, typically with high titre inhibitors and an associated high rate of bleeding. We present data from three patients with MHAI who were successfully treated with Rituximab alone and unequivocally prove that such inhibitors respond to this agent. A treatment protocol is suggested. PMID- 17083519 TI - Normalization of factor VIII levels in a patient with mild haemophilia A during a 35-year period. AB - We report the case of a 45-year-old man who was diagnosed in June 1969 9 years old as having mild haemophilia A following a traumatic left shoulder bleed when his factor VIII (FVIII) activity was 11 IU dL(-1) based on a two-stage assay. The bleed resolved following treatment with intravenous cryoprecipitate. There was no family history of haemophilia. Cryoprecipitate infusions were required to treat further traumatic bleeds between 1971 and 1981. During this time, his FVIII activity was confirmed at 14 IU dL(-1). He defaulted many hospital appointments until 1991 when his FVIII activity had risen to 42 IU dL(-1). There was no evidence of infection, inflammatory or liver disease to account for this change. By 2005 he had a normal FVIII activity of 62 IU dL(-1) based on a one-stage assay. FVIII gene analysis confirmed a codon 531 mutation. It appeared that the discrepant FVIII results related to whether a two-stage or one-stage assay was used that has been previously reported for other patients with these mutations. We felt it important to raise awareness that this phenomenon may lead to apparent correction of haemophilia A. PMID- 17083520 TI - Heterotropic bone formation as a complication of elective joint replacement in haemophilic patients - a case report and literature review. AB - We report two cases of this rare postoperative condition. Both patients having undergone extensive surgery to their hips, developed increased bone growth in the soft tissue of the hip, accounting for the reduced range of movement, pain and stiffness. Plain x-rays taken confirmed the presence of heterotopic ossification, also known as a variant of myositis ossificans. We review the literature and discuss treatment options of this rare condition in patients with end-stage haemophilic arthropathy. PMID- 17083521 TI - Management of acute bleeding in a patient with congenital afibrinogenaemia. AB - Congenital afibrinogenaemia is a rare bleeding disorder characterized by absence of fibrinogen and varying bleeding tendency. Treatment with fibrinogen concentrates is considered to be the best choice for afibrinogenaemic patients who experience bleeding. We report the case of a 22-year-old Greek patient who presented with large muscular haematomas and was treated with fibrinogen concentrates. The efficacy of this treatment and the problems that arose during his hospitalization are being discussed. PMID- 17083522 TI - The feasibility of total ankle prosthesis for severe arthropathy in haemophilia and prothrombin deficiency. AB - The standard treatment for end-stage arthropathy of the ankle joint in haemophilia has been fusion of the ankle joint. Total ankle replacement is used in osteoarthritis and especially in rheumatoid arthritis with good medium-term results. In this case series three patients are being described, in which a total of five total ankle replacements have been preformed. After a median follow up of 4.3 years (range 1-8.7) all prostheses were still in place and did not show any signs of loosening. Clinical scores showed a good to excellent result. In this small series total ankle replacement in patients with bleeding disorders show promising results. Further studies are needed to show the value of this relatively new type of surgery in haemophilic patients. PMID- 17083523 TI - Torticollis as a sign of cervico-thoracic epidural haematoma in an infant with severe haemophilia A. AB - We describe the case of a spinal epidural haematoma in an infant with severe haemophilia A. Initial signs and symptoms were non-specific resulting in delay of the diagnosis and more definitive therapy. The patient eventually developed torticollis, acute flaccid paralysis of the upper extremities, and respiratory distress, prompting radiological examination of the spinal cord. The patient was treated with recombinant FactorVIII and laminectomy. Neurological recovery was complete 3 months following the event. We hypothesize that infants with haemophilia may be at higher risk for this rare complication because of their increasing mobility, frequent falls while cruising furniture, and lack of prophylactic factor replacement. Non-specific signs such as irritability without a focus should alert the clinician to this diagnostic possibility. Torticollis should prompt rapid radiological evaluation of the cervical spine with magnetic resonance imaging to avoid delay in diagnosis. PMID- 17083524 TI - Acquired von Willebrand's syndrome resulting from untreated hypothyroidism in two prepubertal girls. AB - Acquired von Willebrand syndrome (AvWS) is a rare bleeding disorder associated with a number of different diseases, including hypothyroidism. We describe two prepubertal girls with AvWS and undiagnosed hypothyroidism due to thyroiditis Hashimoto. The patients had neither family history nor symptoms of bleeding disorders. Substitution therapy with levothyroxine led to normalization of the coagulation parameters. We report these cases in order to raise awareness among paediatricians so that the AvWS should be suspected and searched for with the appropriate laboratary tests in all cases of hypothyroidism. Moreover, patients with bleeding diathesis of unknown origin should also be investigated for hypothyroidism. PMID- 17083525 TI - Switching between full-length and B-domain-deleted factor VIII and the risk of inhibitors. PMID- 17083526 TI - The Stokes-Einstein equation and the physiological effects of vitreous surgery. PMID- 17083527 TI - Adverse effects of fluorescein angiography. PMID- 17083528 TI - Intravitreal steroids in the management of macular oedema. AB - The use of intravitreal corticosteroids in the management of macular oedema has recently gained widespread acceptance. New long-acting steroid preparations and methods of delivery have facilitated the use of these new modalities. This review describes the various types of macular oedema for which this therapeutic option is used and the results. PMID- 17083529 TI - Paediatric aphakic glaucoma. AB - CONTEXT: Aphakic glaucoma is a serious, sight-threatening complication in children who remain aphakic following congenital cataract surgery. The reported incidence varies from 15% to 45% and it has a higher incidence in small eyes and in babies who undergo surgery before 4 weeks of age. Most cases take the form of open-angle glaucoma. Despite careful monitoring, diagnostic difficulties in children may lead to delayed treatment. Aphakic children require ongoing monitoring, including examinations carried out under general anaesthesia. MANAGEMENT: Contrary to earlier optimism, it is unlikely, according to current clinical information, that intraocular lenses provide protection against the development of glaucoma. It is likely that the lower incidence of glaucoma in pseudophakic eyes results from selection bias for lens insertion. Both medical and surgical treatments have a role in glaucoma management. Trabeculectomy with mitomycin C or Seton implantation (glaucoma drainage device) form the mainstay of surgical treatment. The success rate with Seton implants is better in the short term and more promising in the longer term than that of trabeculectomy. Cyclodestructive procedures play a role in refractory glaucoma. Success rates for surgery range from 14% to 44% and many children require additional medical treatment. Children with aphakic glaucoma need lifelong care. Despite our best efforts, many have poor vision, poor glaucoma control and ultimately become blind. PMID- 17083530 TI - Adverse effects of fluorescein angiography in hypertensive and elderly patients. AB - PURPOSE: To evaluate adverse effects in patients undergoing fundus fluorescein angiography (FA), and to assess the level of these adverse effects in hypertensive and elderly patients. METHOD: We carried out a retrospective study of 358 patients undergoing FA. They were assessed for adverse effects and their relationship to medical history, pre- and post-FA blood pressure and age. RESULTS: The overall adverse reaction rate (AR) was 11.2%, the majority (80%) of which involved nausea. There were no mortalities and no cases of anaphylaxis or cardiac arrest. Hypertension (systolic or diastolic) did not increase the AR. Interestingly, raised initial systolic blood pressure (BP) > or = 160 showed a statistically significant decreased AR compared with that for BP < 160 (7% versus 15%, respectively; p = 0.04). There was no statistical correlation between AR and age. CONCLUSIONS: FA is a relatively safe procedure, but serious side-effects have been reported in the past and therefore adequate measures need to be in place to cope with such emergencies. In our study, there was no increase in the incidence of adverse effects in patients who were hypertensive or elderly. Therefore, these patients should not be denied an important diagnostic procedure that may improve their quality of life purely because of hypertension or age. PMID- 17083531 TI - Photodynamic therapy in subfoveal and juxtafoveal idiopathic and postinflammatory choroidal neovascularization. AB - PURPOSE: To evaluate the efficacy of photodynamic therapy (PDT) in the treatment of idiopathic and inflammatory choroidal neovascularization (CNV). METHODS: This study represents a non-randomized, multicentre, interventional case series. Sixteen eyes of 16 patients with classic or predominantly classic sub- and juxtafoveal idiopathic CNV and 26 eyes of 24 patients with classic or predominantly classic sub- and juxtafoveal CNV secondary to inflammatory conditions were treated with PDT with verteporfin. Best corrected visual acuity (BCVA) before and after PDT, the number of lines gained or lost and the number of PDT sessions were analysed. RESULTS: The mean follow-up periods were 20 months (SD 11) and 16 months (SD 9) for the idiopathic and inflammatory groups, respectively. The mean BCVA in idiopathic CNV changed from 0.63 logMAR (SD 0.37) before treatment to 0.82 logMAR (SD 0.43) after treatment (p = 0.16; Student's t test paired data). The number of treatments performed averaged 2.1 (range 1-4, SD 0.9). The mean BCVA in postinflammatory CNV changed from 0.60 logMAR (SD 0.4) before treatment to 0.57 logMAR (SD 0.6) after treatment (p = 0.79; Student's t test paired data). The mean number of treatments was 2.6 (range 1-6, SD 1.2). CONCLUSIONS: Photodynamic therapy with verteporfin may be useful in stabilizing BCVA in patients with sub- and juxtafoveal postinflammatory CNV and subfoveal idiopathic CNV. PMID- 17083532 TI - Trans-Tenon's retrobulbar triamcinolone infusion for myopic choroidal neovascularization. AB - PURPOSE: To report the effects of trans-Tenon's retrobulbar injection of triamcinolone acetonide for subfoveal and juxtafoveal choroidal neovascularization (CNV) caused by pathological myopia. METHODS: Eleven consecutive patients (11 eyes) with myopic CNV were treated with trans-Tenon's retrobulbar injection of triamcinolone acetonide. Each patient received a single injection. Evaluation included best corrected visual acuity (BCVA) measurements, fluorescein fundus angiography, retinal oedema examined by optical coherence tomography (OCT), and retinal sensitivity using scanning laser ophthalmoscopy (SLO) at the initial examination and at 6 and 12 months after treatment. RESULTS: At 6 months after treatment, BCVA had improved by at least two ETDRS lines in eight eyes, and remained unchanged in three eyes. No eye showed worsening of VA by two or more ETDRS lines. At 12 months, BCVA had improved by at least two ETDRS lines in 10 eyes and remained unchanged in only one eye. The size of the CNV decreased in all patients after treatment. Fluorescein fundus angiography revealed an absence of dye leakage in the late angiographic phase. Optical coherence tomography revealed decreased retinal oedema in all patients and SLO microperimetry revealed an increase in retinal sensitivity in seven eyes, at both 6 and 12 months after treatment. Chorioretinal atrophy developed around the CNV in 10 eyes at 6 months and in all eyes at 12 months after treatment. CONCLUSIONS: Trans-Tenon's retrobulbar injection of triamcinolone acetonide for CNV resulting from pathological myopia appears to be relatively safe and to have a good visual outcome, although a longterm follow-up study in a larger series of patients is necessary. PMID- 17083533 TI - 33 cases of airsoft gun pellet ocular injuries in Copenhagen, Denmark, 1998-2002. AB - PURPOSE: To evaluate the incidence and ocular effects of blunt trauma due to injury from airsoft gun pellets. METHODS: We conducted a non-comparative case series based on the files of 33 patients who suffered ocular injury from airsoft guns and were admitted to one university emergency eye clinic in Copenhagen during a 5-year period. RESULTS: A total of 33 eyes in 33 patients were examined. Thirty male and three female patients were affected. Mean age was 13 years (range 3-49 years). Mean follow-up time was 6.5 days (range 1-540 days). On initial examination, we found: hyphaema (n = 28), corneal abrasion (n = 22), retinal oedema (n = 11), subconjunctival haemorrhage (n = 10), palpebral haemorrhage and/or oedema (n = 9), iris dialysis (n = 7), intraocular pressure (IOP) > 31 mmHg (n = 4), IOP < 9 mmHg (n = 4), vitreous haemorrhage (n = 2) and cataract (n = 1). One patient had brown corneal discoloration due to severe chronic hyphaema that required surgery. The final visual acuity was 1.0 in 20 patients and < or = 0.7 in five patients. CONCLUSION: The airsoft gun trauma resulted in anterior and posterior segment ocular injuries that required acute medical intervention and for which some patients were hospitalized. Trauma mainly occurred in children and youths. A longterm risk of glaucoma is expected. Eye care professionals and parents as well as manufacturers of airsoft guns should advise participants to wear adequate protection when involved in this activity. PMID- 17083535 TI - Risk factors for endothelial cell loss post-keratoplasty. AB - PURPOSE: Transplant survival following penetrating keratoplasty is determined to a large extent by the course of endothelial cell density loss. Different influencing factors such as organ culture conditions, surgical trauma, exchange between donor and recipient cells, cell ageing and immune reactions can contribute to endothelial cell loss. The aim of this study was to determine the rate of endothelial cell loss in our patients and to detect dependencies on donor and recipient-related factors. METHODS: Using non-contact specular microscopy, endothelial cell counts were obtained every 6 months from 293 consecutive patients who underwent keratoplasty in our institution between 1996 and 2000. Follow-up time was 36 months. RESULTS: In comparison with the density of donor endothelial cells, the mean endothelial cell loss of patients was 28.8% after 6 months, 39.8% after 12 months and 49% after 24 months. Donor age and initial cell density did not have a significant influence on the course of endothelial cell loss. The lowest rate of endothelial cell loss was associated with patients diagnosed with keratoconus. Conversely, those with preoperative glaucoma had a significantly increased rate of endothelial cell loss (p < 0.05). CONCLUSIONS: This study shows that preoperative glaucoma is a major risk factor for increased endothelial cell loss following keratoplasty. PMID- 17083534 TI - Simultaneous quantification of 17 immune mediators in aqueous humour from patients with corneal rejection. AB - PURPOSE: To simultaneously quantitate and compare the concentrations of 17 immune mediators: (1) the cytokines interleukin-1beta, IL-2, IL-4, IL-5, IL-6, IL-7, IL 10, IL-12p70, IL-13, IL-17, tumour necrosis factor-alpha, interferon-gamma; (2) the growth factors granulocyte-monocyte colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF), and (3) the chemokines CXCL-8, monocyte chemoattractant protein-1, and macrophage inflammatory protein-1beta in aqueous humour from patients with corneal rejection and patients with a non inflammatory condition in the anterior chamber. METHODS: Aqueous humour was obtained by paracentesis of the anterior chamber in 14 patients with corneal rejection, three patients with cataract and six patients with Fuchs' endothelial dystrophy. Simultaneous quantitation of 17 mediators in 25 micro l aqueous humour from each patient was performed by employing a highly sensitive Luminex 100 multiplex array assay. RESULTS: All 17 immune mediators were detected in aqueous humour from rejection patients. The ranges of the immune mediators were determined. The immune mediators were significantly increased in aqueous humour from rejection patients compared with that from other patients. CONCLUSIONS: The Luminex 100 multiplex array assay is very efficient in simultaneous quantitation of multiple immune mediators in small volumes of aqueous humour. A total of 17 immune mediators were increased in aqueous humour from rejection patients. This underlines the complex immunological interactions of the rejection process. PMID- 17083536 TI - Automated lamellar therapeutic keratoplasty (ALTK) in the treatment of anterior to mid-stromal corneal pathologies. AB - PURPOSE: To evaluate the efficacy of automated lamellar therapeutic keratoplasty (ALTK) in the treatment of anterior to mid-stromal corneal pathologies. METHODS: A prospective clinical study was undertaken in 15 eyes of 15 patients who underwent ALTK using the ALTK System (Moria/Microtek Inc., Doylestown, Pennsylvania, USA). The parameters evaluated were uncorrected and best corrected visual acuity (BCVA), keratometry, pachymetry, contrast sensitivity, glare acuity, intraoperative surgical time and complications, postoperative time to epithelialization, graft clarity and postoperative complications. RESULTS: The mean surgical time was 46 +/- 9 mins. The BCVA, contrast sensitivity and glare acuity significantly improved at 6 months postoperatively. The mean decimal BCVA was 0.08 +/- 0.03 preoperatively, which improved to 0.33 +/- 0.30 at 6 months, when 11 eyes had BCVA > or = 6/18. The median epithelialization time was 3 days (range 1-10 days). CONCLUSIONS: Automated lamellar therapeutic keratoplasty appears to be safe and effective for diseases affecting the anterior to mid stroma of the cornea. PMID- 17083537 TI - Donor-derived myofibroblasts in the ocular surface after allogeneic haematopoietic stem cell transplantation. AB - PURPOSE: To identify and characterize cells of donor origin in the ocular surface of female recipients who have undergone allogeneic haematopoietic stem cell transplantation (allo-SCT) from a male donor. METHODS: Cytological impressions from the eyes of nine allografted patients (17 eyes) were analysed. Donor cells were identified using sex-chromosome-specific fluorescence in situ hybridization (FISH). Cells were characterized by immunohistochemistry (IHC) using the CK3 and CK19 epithelial markers, the panleucocytic marker CD45 and the myofibroblast marker alpha-SMA. RESULTS: No epithelial cells of donor origin were observed in the corneal or conjunctival samples. Cells of donor origin were found in the corneal samples, although these were often too degraded to allow characterization by IHC. In the conjunctiva, a median of 86% of the total number of cells were of recipient origin, including a subgroup (2%) of giant cells exhibiting polyploidy (range 4-18 n), found in the limbal region. Donor cells were detected in the conjunctiva of all nine patients at a median ratio of 9%, of which two-thirds were CD45+/alpha-SMA+. CONCLUSIONS: We observed superficially located myofibroblasts of donor origin in all allografted patients, but not in samples from healthy controls. Whether myofibroblasts are implicated in ocular graft versus-host disease requires further studies. PMID- 17083538 TI - Ocular findings in four children with mucopolysaccharidosis I-Hurler (MPS I-H) treated early with haematopoietic stem cell transplantation. AB - PURPOSE: To present visual functions and ocular findings in four children with mucopolysaccharidosis I-Hurler (MPS I-H) treated early with stem cell transplantation (SCT). METHODS: Clinical ophthalmological evaluations including visual evoked potentials (VEPs) were carried out. RESULTS: Stem cell transplantation was performed before 20 months of age. Ocular follow-up lasted 1.3-5.6 years (median 4.1 years). Reductions in corneal opacities were observed in all four children post-SCT, but a slight cloudiness persisted. Decreased visual acuity and high hyperopia (median + 6.25 dioptres, range + 4.0 D to + 7.5 D spherical equivalents) were noted in all children. Hyperopia was initially undetected due to dull retinal reflexes and photophobia. Two children developed esotropia, one with amblyopia. Keratometry, performed in two children, demonstrated subnormal values with a mean of 39.33 D (range 37.62-41.00 D). Visual evoked potentials and intraocular pressures were normal. Neither cataract nor dry eye were detected during follow-up. CONCLUSIONS: Early SCT appears to be beneficial in reducing, but not eliminating, corneal opacities in children with MPS I-H. Subjects are at risk of developing high hyperopia and esotropia. Hyperopia might be caused by the storage of glucosaminoglucans that increase corneal rigidity, thereby straightening the curvature of the cornea and reducing refractive power. As early diagnosis and treatment are very important, paediatric ophthalmologists should remember to rule out MPS I-H in children with corneal opacities. PMID- 17083539 TI - Use of amniotic membrane as an adjuvant in refractory glaucoma. AB - PURPOSE: To evaluate the results after amnion-shielded trabeculectomy with concomitant use of mitomycin C. METHODS: The study comprised patients with severely refractory glaucomas who had previously undergone two or more regular trabeculectomies with mitomycin C and, in one case, cyclodestructive cryotherapy. The patients were admitted for surgery between May 2003 and May 2004. Trabeculectomy was performed through a limbus-based incision in nine eyes of nine patients. Cryopreserved human amniotic membranes obtained from our biobank were impregnated with 0.4 mg/ml mitomycin C and washed for 2 mins in balanced salt solution. One membrane was sutured to the scleral surface and partly positioned under the scleral flap, and another was secured to subconjunctival tissue. RESULTS: The mean preoperative intraocular pressure (IOP) was 32.2 mmHg (range 22 44 mmHg). After a follow-up of 6-18 months (mean 9.8 months), the mean IOP was 16.4 mmHg (range 11-26 mmHg). The mean number of glaucoma medications was reduced from 2.4 preoperatively to 1.4 postoperatively. There were no devastating complications. CONCLUSIONS: Amnion-shielded trabeculectomy is a procedure that should be evaluated as an option in refractory glaucoma. PMID- 17083540 TI - Optical coherence tomography versus automated perimetry for follow-up of optic neuritis. AB - PURPOSE: To evaluate the usefulness of optical coherence tomography (OCT) for follow-up of optic neuritis (ON) compared with subjective visual function tests. METHODS: Twelve patients with ON underwent a complete ophthalmological evaluation at initial diagnosis, including best corrected visual acuity (VA), visual fields testing and OCT examination. These examinations were repeated periodically over 6 months. Retinal nerve fibre layer (RNFL) thickness evolution was analysed and correlated with VA and visual field mean deviation. RESULTS: Six months after ON, mean RNFL in the affected eye decreased significantly compared with that in the fellow eye (24.54%). A significant correlation was found between RNFL thinning and final VA (r = 0.807, p = 0.005), with a 1-line drop in VA for every 5.4-micro m decrease. All patients with an altered visual field had an abnormal RNFL value; of the seven patients with normal visual fields, 57% had an abnormal RNFL value (p < 5%). CONCLUSIONS: Optical coherence tomography can detect axonal damage as early as the third month after an isolated initial episode of ON, even in the presence of normal visual fields. Mean RNFL thinning is correlated with final VA. PMID- 17083541 TI - The relationship between stress and acute anterior uveitis. AB - PURPOSE: To examine the relationship between stress and anterior uveitis using the General Health Questionnaire (GHQ) and the Social Readjustment Rating Questionnaire (SRRQ). METHODS: Patients attending eye casualty with acute anterior uveitis (AAU) and suitable controls were asked to complete the GHQ and the SRRQ. A follow-up postal survey, using the same questionnaires, was repeated at least 3 months later amongst AAU patients only. Scores for the GHQ and SRRQ at initial presentation and at follow-up were compared for different groups. RESULTS: Over 12 months, 42 patients and 25 controls participated in the first stage of this study. Patients with AAU had higher GHQ scores than controls (mean 6.8 versus 3.2, p = 0.01). A total of 25 patients responded to the follow-up postal survey. At follow-up, 13 had experienced resolution and 12 had recurrence. In the group with resolution, there was a significant fall in GHQ scores (mean 6.1 versus 1.5, p = 0.0044). Patients with recurrence had higher GHQ scores than those with resolution (mean 6.8 versus 1.5, p = 0.02). CONCLUSIONS: Using the GHQ reveals a clear relationship between stress and the recurrence of AAU in susceptible individuals. PMID- 17083542 TI - The key role of electrophysiology in the diagnosis of visually impaired children. AB - PURPOSE: To describe the outcome of specialized electrophysiology in visually impaired children. METHODS: We carried out a retrospective evaluation of 340 electrophysiological examinations performed in 298 children over a 3-year period (2001-2003), with regard to demographic data, referral pattern, degree of compliance, and diagnostic results. Electrophysiology was performed without sedation or anaesthesia. In electroretinograms, DTL electrodes were used in combination with online selection of responses. Visual evoked potentials testing was performed with seven active occipital electrodes. RESULTS: The mean age of the children was 7 +/- 5 years; 72 (24%) of the children were mentally as well as visually impaired. Main reasons for referral were suspected posterior segment disease, abnormal visual development, unexplained low vision, high myopia, and suspected albinism. Compliance was good in 302/340 (88%), partial in 24/340 (7%), and absent in 14/340 (4%) of the examinations. Of the 326 successful procedures, 215 (66%) showed abnormal results. Tapetoretinal dystrophy (22%), opticopathy (16%), congenital stationary night blindness (13%), and cone dystrophy (11%) were the most frequently established diagnoses. Albinism was confirmed in 14 of 24 suspected patients; additionally, unsuspected misrouting was found in six. In 26 (9%) of the patients, a previously established diagnosis was changed. CONCLUSIONS: In a specialized setting, electrophysiological examinations can be performed successfully in visually impaired children. The results are essential for the final ophthalmological diagnosis and have important consequences for rehabilitation. PMID- 17083543 TI - Sensitivity and specificity of a visual acuity screening protocol performed with the Lea Symbols 15-line folding distance chart in preschool children. AB - PURPOSE: The aim of this study was to assess the feasibility of a visual acuity (VA) test using the Lea Symbols 15-line folding distance chart and its diagnostic validity in detecting VA deficiency in preschool children. METHODS: A group of 149 children aged 38-54 months underwent VA examination performed with the Lea 15 line folding optotype at a distance of 3 metres, according to a test protocol described in the Methods section. After the VA test, a complete ophthalmological examination, including cycloplegic retinoscopy, a cover test and examination of the anterior and posterior segments, was performed on each child in order to detect any VA-threatening ocular abnormality. The Lea Symbols test's sensitivity, specificity, positive and negative likelihood ratios (LR +, LR -) and the receiver operating characteristic (ROC) curve were calculated by means of standard procedures using each VA level of the chart from 0.1 to 1 (1-0 logMAR) as a cut-off point. RESULTS: The Lea Symbols test could be successfully used in 95.9% of the population. The most useful cut-off points for screening preschool children were found to be 0.8 (LR + 5.73, LR - 0.05) or 0.63 (LR + 11.7, LR - 0.23). CONCLUSION: The Lea Symbols test proved to be clinically useful in detecting VA deficiency in preschool children. The choice between the two best performing cut-off levels should be made according to the expected cost effectiveness of the screening programme. PMID- 17083544 TI - Vitrectomy for non-ischaemic macular oedema in retinal vein occlusion. AB - PURPOSE: To evaluate the effect of vitrectomy in eyes with non-ischaemic macular oedema secondary to hemi and central retinal vein occlusion. METHODS: This retrospective study analysed the outcome of eight patients with non-ischaemic macular oedema without posterior vitreous detachment. Six patients had a central retinal vein occlusion and two had a hemi retinal vein occlusion. A standard three-port vitrectomy was performed in all patients. Retinal mapping by optical coherence tomography and visual acuity (VA) testing were performed before vitrectomy and at 1, 2 and 12 months postoperatively. RESULTS: At the 1-month follow-up there was a statistically significant reduction in retinal thickness (Wilcoxon; p = 0.04) that persisted at 2 months (Wilcoxon; p = 0.04). However, at 12 months there was no difference compared with baseline. LogMAR VA was significantly improved at 1 month (Wilcoxon p = 0.04), but at 2 and 12 months there was no difference compared with baseline. CONCLUSIONS: Vitrectomy in hemi and central retinal vein occlusion has the potential to reduce macular oedema and improve VA in the early postoperative phase but does not seem to improve the longterm outcome of the disease. PMID- 17083545 TI - Isolated bilateral optic neuritis in acute disseminated encephalomyelitis. AB - PURPOSE: We present a 4-year-old girl with isolated visual loss related to a bilateral oedematous optic neuropathy of unknown origin, occurring after an episode of viral illness. METHODS: A cerebral MRI was performed, showing bilateral enhancement of the optic nerves and bilateral, diffuse, cerebral white matter lesions. RESULTS: The MRI scan suggested acute disseminated encephalomyelitis. Treatment with intravenous steroids and immunoglobulins resulted in favourable visual recovery. CONCLUSIONS: Bilateral isolated optic neuritis in childhood may be the only presenting symptom in acute disseminated encephalomyelitis, which requires appropriate evaluation and therapeutic management. PMID- 17083546 TI - Iris microhaemangiomas and idiopathic juxtafoveolar retinal telangiectasis. AB - We describe a case of iris microhaemangiomas (iris vascular tufts) causing spontaneous hyphaema and where examination showed that the patient also had idiopathic juxtafoveolar retinal telangiectasis. A systematic search through PubMed, EMBASE and reference lists revealed a total of 90 reported cases of iris microhaemangiomas. To our knowledge, no previous reports have described concurrent findings of iris microhaemangiomas and idiopathic juxtafoveolar retinal telangiectasis in a patient. Both conditions are acquired vascular diseases, but their pathogenesis and aetiologies are unknown. Iris microhaemangiomas are important clinically because they are occasionally a cause of spontaneous hyphaema and may induce transient elevation of intraocular pressure. PMID- 17083547 TI - Abnormal optic discs and renal failure: papillorenal syndrome. PMID- 17083548 TI - Vital staining of squamous cell carcinoma of the conjunctiva using toluidine blue. PMID- 17083549 TI - Five-year follow-up of posterior capsule opacification with two different silicone intraocular lenses. PMID- 17083550 TI - Deposition of triamcinolone crystals on the posterior lens capsule following prone posturing post-vitrectomy. PMID- 17083551 TI - An extreme 8.0-dioptre myopic shift with malignant glaucoma. PMID- 17083552 TI - Resolution of diabetic macular oedema following high altitude exercise. PMID- 17083553 TI - Photodynamic therapy with verteporfin combined with intravitreal injection of bevacizumab for exudative age-related macular degeneration. PMID- 17083554 TI - Retinal pigment epithelial tear following intravitreal bevacizumab injection for neovascular age-related macular degeneration. PMID- 17083555 TI - Intravitreal bevacizumab (Avastin) injection associated with regression of retinal neovascularization caused by sickle cell retinopathy. PMID- 17083556 TI - Choroidal neovascularization treated with intravitreal injection of bevacizumab (Avastin) in angioid streaks. PMID- 17083557 TI - Measuring patients' preferences for participating in health care decisions: avoiding invalid observations. PMID- 17083558 TI - Inside the black box of shared decision making: distinguishing between the process of involvement and who makes the decision. AB - BACKGROUND: Shared decision making has practical implications for everyday health care. However, it stems from largely theoretical frameworks and is not widely implemented in routine practice. AIMS: We undertook an empirical study to inform understanding of shared decision making and how it can be operationalized more widely. METHOD: The study involved patients visiting UK general practitioners already well experienced in shared decision making. After these consultations, semi-structured telephone interviews were conducted and analysed using the constant comparative method of content analysis. RESULTS: All patients described at least some components of shared decision making but half appeared to perceive the decision as shared and half as 'patient-led'. However, patients exhibited some uncertainty about who had made the decision, reflecting different meanings of decision making from those described in the literature. A distinction is indicated between the process of involvement (option portrayal, exchange of information and exploring preferences for who makes the decision) and the actual decisional responsibility (who makes the decision). The process of involvement appeared to deliver benefits for patients, not the action of making the decision. Preferences for decisional responsibility varied during some consultations, generating unsatisfactory interactions when actual decisional responsibility did not align with patient preferences at that stage of a consultation. However, when conducted well, shared decision making enhanced reported satisfaction, understanding and confidence in the decisions. CONCLUSIONS: Practitioners can focus more on the process of involving patients in decision making rather than attaching importance to who actually makes the decision. They also need to be aware of the potential for changing patient preferences for decisional responsibility during a consultation and address non-alignment of patient preferences with the actual model of decision making if this occurs. PMID- 17083559 TI - Putting informed and shared decision making into practice. AB - OBJECTIVE: To investigate the practice, experiences and views of motivated and trained family physicians as they attempt to implement informed and shared decision making (ISDM) in routine practice and to identify and understand the barriers they encounter. BACKGROUND: Patient involvement in decision making about their health care has been the focus of much academic activity. Although significant conceptual and experimental work has been done, ISDM rarely occurs. Physician attitudes and lack of training are identified barriers. DESIGN: Qualitative analysis of transcripts of consultations and key informant group interviews. SETTINGS AND PARTICIPANTS: Six family physicians received training in the ISDM competencies. Audiotapes of office consultations were made before and after training. Transcripts of consultations were examined to identify behavioural markers associated with each competency and the range of expression of the competencies. The physicians attended group interviews at the end of the study to explore experiences of ISDM. RESULTS: The physicians liked the ISDM model and thought that they should put it into practice. Evidence from transcripts indicated they were able to elicit concerns, ideas and expectations (although not about management) and agree an action plan. They did not elicit preferences for role or information. They sometimes offered choices. They had difficulty achieving full expression of any of the competencies and integrating ISDM into their script for the medical interview. The study also identified a variety of competency-specific barriers. CONCLUSION: A major barrier to the practice of ISDM by motivated physicians appears to be the need to change well established patterns of communication with patients. PMID- 17083560 TI - Has patients' involvement in the decision-making process changed over time? AB - OBJECTIVE: To get insight into the changes over time of patients' involvement in the decision-making process, and into the factors contributing to patients' involvement and general practitioners' (GPs) communication related to the Medical Treatment Act (MTA) issues: information about treatment, other available treatments and side-effects; informed decision making; asking consent for treatment. BACKGROUND: Societal developments have changed the doctor-patient relationship recently. Informed decision making has become a central topic. Patients' informed consent was legalized by the MTA (1995). DESIGN: Data of two cross-sectional studies, the First (1987) and Second (2001) Dutch National Survey of General Practice, were compared. SETTING AND PARTICIPANTS: General practice consultations; 16 GPs and 442 patients in 1987; 142 GPs and 2784 patients in 2001. METHODS: Consultations were videotaped and rated using Roter's Interaction Analysis System and observer questionnaires; pre- and post-consultation patient questionnaires; and GP questionnaires. Descriptive analyses and multivariate, multilevel analysis were applied. MAIN RESULTS: Most patients reported to have received the information they had considered as important prior to the consultation. There were discrepancies in involvement in treatment decisions and in giving information about other available treatments, side-effects and risks. GPs who were more affective and gave more information, more often involved their patients, especially younger patients, in decision making. In 2001, more informed decision making was observed and the GPs asked consent for a treatment more often, but they less often asked for the patients' understanding. CONCLUSION: Patients' involvement in decision making has increased over time, but not in every respect. However, this does not apply for all patients, especially the older ones. It should be questioned whether they are willing or capable to be involved and if so, how they could be encouraged. PMID- 17083561 TI - Expectations of patients and parents of children with asthma regarding access to complementary therapy information and services via the NHS: a qualitative study. AB - OBJECTIVE: To explore the expectations of patients and parents of children with asthma regarding access to complementary therapies via the NHS. METHODS: Fifty semi-structured interviews with adults and parents of children with asthma, from a range of health-care settings, including users and non-users of complementary therapies. Interviews were recorded, transcribed verbatim and the data were analysed thematically. RESULTS: Thirty-one patients were using complementary therapies for asthma, six were using complementary therapies for other health problems and 13 were non-users. Various therapies were used for asthma, most commonly homeopathy and breathing techniques, predominantly outside the NHS. Two broad themes emerging from the data were expectations about access to information and knowledge about complementary therapies via NHS health professionals, and expectations regarding access to complementary therapy services via the NHS. As a minimum, the majority of participants wanted NHS health professionals to be more 'open' towards and know more about complementary therapies than their patients - perceived as not currently usual. Most were positive about greater NHS access to complementary therapy services, for enhancing patient choice, improving equality in access for less affluent patients and facilitating patients' self-help. Participants who were highly sceptical about complementary therapies argued that lack of scientific evidence of effectiveness prohibited the need for greater complementary therapy knowledge or service provision within the NHS. Alongside their expectations, patients and parents expressed realistic views about facilitators and barriers to greater access. CONCLUSIONS: While health service planners and providers often express reservations about the value of complementary therapies, it is important to take patients' preferences into account if policy discourses regarding patient-centred care and choice are to be realized in practice. PMID- 17083562 TI - Information in mental health: qualitative study of mental health service users. AB - BACKGROUND: Despite the widespread proliferation of consumer health information provision, little is known about information needs or information-seeking behaviour in mental health. A qualitative study was therefore undertaken to explore these issues for mental health service users. DESIGN: In-depth interview study with purposive sample of 36 men and women with experience of mental health problems. RESULTS: Four main themes were identified. A general lack of information was equated with a lack of respect. People undertook their own research into their condition, and recognized the challenge to professionals. Stigma was widespread and inhibited information seeking. There was a desire for an explanation of mental health problems in physical terms. People particularly valued hearing other people's experience of mental health problems, for reasons of universality, instillation of hope, and understanding and empathy. CONCLUSIONS: The findings provide support for a more equal partnership between patients and professionals. Information providers and health practitioners should take account of the value of other people's experience as an information source. PMID- 17083563 TI - Multichannel communication: new opportunities to maximize access to support for people affected by cancer. PMID- 17083564 TI - Loss or down-regulation of HLA class I expression at the allelic level in freshly isolated leukemic blasts. AB - Loss or down-regulation of human leukocyte antigen (HLA) class I expression has been demonstrated in a variety of solid tumors. To date, such altered HLA expression has not been studied extensively in freshly isolated leukemic blasts. If it occurs, leukemic cells could escape T-cell surveillance as a consequence. Genotypes of nine leukemic cell lines were determined using a polymerase chain reaction for HLA classes I and II. Cells were also examined for HLA beta2 microglobulin, and allele-specific HLA protein expression using flow cytometry. Next, 44 samples of freshly isolated leukemic blasts from 43 patients with malignant hematological diseases were examined for allele-specific HLA expression using flow cytometry. Microsatellite analysis was performed to determine heterozygosity in the HLA region on chromosome 6. Genotype analysis for HLA class I together with microsatellite analysis demonstrated loss of HLA haplotype in HL 60 cells. No loss of HLA haplotype was observed in 44 samples of freshly isolated leukemic blasts. As reported previously, flow cytometric analysis rarely demonstrated loss or down-regulation of HLA expression at initial diagnosis (3/39; 7.7%); however, this was evident in two of five cases in relapse (40.0%), which contrasts with previous reports. In one patient with acute leukemia, HLA-A2 cell surface expression was present at initial diagnosis, lost at relapse, and completely restored after 48 h of culture in the presence of interferon-gamma. These results suggest loss of allele-specific HLA expression may be involved in the pathogenesis of relapse in patients with leukemia. The findings should be valuable in designing new strategies for clinical immunotherapy. PMID- 17083565 TI - Evaluation of S100A10, annexin II and B-FABP expression as markers for renal cell carcinoma. AB - This study aimed to analyze expression of S100A10, annexin II and B-FABP genes in renal cell carcinoma (RCC) and their potential value as tumor markers. Furthermore, any correlation between the gene expression and prognostic indicators of RCC was analyzed. Expression of each gene was estimated by RT-PCR in the non-neoplastic (normal) and tumorous parts of resected kidney samples. Also, each antigen was immunostained in RCC and normal kidney tissues. Expression of the S100A10 gene averaged 2.5-fold higher in the tumor than that in the normal tissues (n = 47), after standardization against that of beta-actin. However, expression of annexin II, a natural ligand of S100A10, was only 1.64-fold higher. In the tissue sections of RCC, S100A10 and annexin II were immunostained in membranes. In the normal renal epithelia, however, both antigens were stained in the Bowman's capsule and the tubules from Henle's loop through the collecting duct system, but not in the proximal tubules, from where most RCC are derived. In contrast, expression of the B-FABP gene was 20-fold higher in the tumor. No B FABP was immunohistochemically detected in normal kidney sections, but it was stained in the cytoplasm of RCC tissue sections. S100A10 and B-FABP genes were overexpressed regardless of nuclear grade and stage of RCC. Immunopositivity in RCC tissues (n = 13) was 100% for S100A10 and annexin II, and 70% for B-FABP; however, no clear relationship was observed in either antigen with nuclear grade and stage. It was found that all three performed well as RCC markers. B-FABP was most specific to RCC, as it was expressed little in normal kidney tissues. PMID- 17083566 TI - High JC virus load in gastric cancer and adjacent non-cancerous mucosa. AB - The JC virus (JCV) infects a large proportion of the worldwide population and approximately 90% of adults are seropositive. Recent reports have described the possibility of its oncogenetic role in several malignancies. The aim of the present study was to assess the oncogenetic significance of JCV for gastric cancer. Twenty-two sample pairs of fresh tumor and adjacent non-cancerous tissue (ANCT) as well as 10 normal gastric mucosa specimens were investigated on the basis of nested polymerase chain reaction (PCR) followed by Southern blotting, DNA direct sequencing, real-time PCR, in situ PCR and immunohistochemistry. The T antigen sequence was detected in 86.4% of gastric cancers and ANCT, and in 100% of the normal mucosa samples, as for virus capsid protein, 54.1%, 68.1% and 70%, respectively. A generally low incidence was noted for agnoprotein. The JCV DNA load was approximately 10-fold higher in both gastric cancers and paired ANCT (4784 +/- 759 and 5394 +/- 1466 copies/microg DNA, respectively) than in normal gastric tissue (542.4 +/- 476.0 copies/microg DNA, P < 0.0001). In situ PCR revealed sporadic JCV genome-positive cancer cells and foveolar epithelial cells. T antigen protein expression assessed by immunohistochemistry was detected only in one case (1/22; 4.5%), probably because the half life of T antigen might be short. It was concluded that the gastric epithelium in most Japanese people is infected with JCV at a low rate but levels of infection are increased markedly in both cancer cells and ANCT, indicating that multiplication of JCV copies might be a risk factor and a background for gastric carcinogenesis. PMID- 17083567 TI - Hedgehog signal pathway is activated in ovarian carcinomas, correlating with cell proliferation: it's inhibition leads to growth suppression and apoptosis. AB - The hedgehog (Hh) signal pathway has recently been shown to be activated in human malignancies. However, little is known about its role in the development or patient prognosis of epithelial ovarian carcinoma. In the present study, we examined in vivo and in vitro the expression and functional role of Hh signal molecules in epithelial ovarian tumors and normal ovarian surface epithelial (OSE) cells. The expression of Shh, Dhh, Ptch, Smo and Gli1 proteins was not observed in normal OSE, but was increased stepwise in benign, borderline and malignant neoplasms. In addition, immunoreactivity for Shh, Dhh, Ptch, Smo and Gli1 was highly correlated with cell proliferation assessed by Ki-67. Blocking the Hh signal using either the Hh pathway inhibitor cyclopamine or Gli1 siRNA led to remarkably decreased cell proliferation in ovarian carcinoma cells. Treatment with cyclopamine induced not only G, arrest but also apoptosis along with the downregulation of cyclin A and cyclin D1, and the upregulation of p21 and p27. Among the Hh signal molecules, Dhh expression was correlated with poor prognosis of ovarian carcinoma patients. These findings suggest that the Hh signal pathway plays an important role in ovarian tumorigenesis as well as in the activation of cell proliferation in ovarian carcinomas. Thus, the Hh signal pathway is a possible molecular target of new treatment strategies for ovarian carcinoma. PMID- 17083568 TI - Increased susceptibility to hepatocarcinogenicity of Nrf2-deficient mice exposed to 2-amino-3-methylimidazo[4,5-f]quinoline. AB - To elucidate the roles of the transcription factor NF-E2-related factor (Nrf2) in hepatocarcinogenesis induced by 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), a mutagenic and carcinogenic heterocyclic amine, Nrf2-deficient mice were treated with 300 p.p.m. IQ in their diet for 1, 4 or 52 weeks. In the long-term experiment, the multiplicity and incidence of liver tumors in male and female IQ treated Nrf2 deficient (-/-) mice were significantly higher than those in their counterpart wild-type (+/+) mice exposed to IQ. In the short-term experiment, although IQ exposure to Nrf2(+/+) mice of both sexes did not modify UDP glucuronosyltransferase values, glutathione S-transferase values were significantly increased due to IQ treatment, in contrast to no alteration in male and female Nrf2(-/-) mice. Levels of oxidative stress markers such as 8 hydroxydeoxyguanosine and thiobarbituric acid reactive substances in the livers of all treated mice were not changed by IQ treatment. IQ-specific DNA adduct levels were elevated only in female Nrf2(-/-) mice, although the increase was not significant. IQ treatment caused an increase in proliferating cell nuclear antigen labeling indices only in male Nrf2(-/-) mice. The present data clearly show that Nrf2(-/-) mice of both sexes are susceptible to IQ hepatocarcinogenicity, which might result from IQ accumulation due to failure of metabolizing enzyme induction. In addition, inconsistent results concerning IQ specific adducts and proliferating cell nuclear antigen labeling indices in male and female Nrf2(-/-) mice suggest the existence of different contributions of Nrf2 to IQ hepatocarcinogenesis between mice of the two sexes. PMID- 17083569 TI - Identification of DFNA5 as a target of epigenetic inactivation in gastric cancer. AB - Epigenetic gene inactivation plays a key role in the development of various types of cancer. Using methylated CpG island amplification coupled with representational difference analysis to identify genes inactivated by DNA methylation in gastric cancer, we identified seven DNA fragments corresponding to the 5' CpG islands of the affected genes. One of the clones recovered was identical to the 5' flanking region of DFNA5, a gene previously shown to be associated with deafness and induced by DNA damage. Further analysis revealed that DFNA5 is expressed in normal tissues but is down-regulated in gastric cancer cell lines due to methylation of the region around its transcription start site. Treating gastric cancer cells that lacked DFNA5 expression with a methyltransferase inhibitor, 5-aza-2'-deoxycytidine, restored the gene's expression. Methylation of DFNA5 was detected in 50% of primary gastric tumors, and was correlated with positivity for Epstein-Barr virus and the absence of metastasis. Moreover, introduction of exogenous DFNA5 into silenced cells suppressed colony formation. Taken together, these data suggest that the silencing of DFNA5 occurs frequently in gastric cancer and may play a key role in development and progression of the disease. PMID- 17083570 TI - Equine sweating and anhidrosis Part 1--equine sweating. AB - Sweating has a variety of functions in mammals including pheromone action, excretion of waste products and maintenance of the skin surface ecosystem. In a small number of mammalian species, which includes humans and the Equidae, it also has an important role in thermoregulation. This review is focused specifically on the thermoregulatory role of sweat in Equidae and the causes of sweating failure (anhidrosis). The first part describes the glandular appearance, sweat composition, and output rates; and considers the latest theories on the glandular control and secretory mechanisms. It is concluded that the glands are not directly innervated but are controlled by the interplay of neural, humoral and paracrine factors. The secretory mechanism is not as simple as previously thought and is mediated by the dynamic interaction of activating pathways, including autocrine control not only of the secretory process but probably also of secretory cell reproduction, growth, and death. PMID- 17083571 TI - An overview on congenital alopecia in domestic animals. AB - Alopecia, that is, lack of hair in any quantity, is a frequent complaint of pet owners. Although mostly acquired, rare congenital forms of alopecia exist that are associated with abnormalities in hair follicle morphogenesis. Congenital alopecias can result in changes in quality or quantity of hair follicles and the hair fibres produced by them. They vary in terms of clinical presentation and mode of inheritance. Histopathology is usually needed in order to differentiate between a reduced number of otherwise normal hair follicles and qualitative hair follicle abnormalities. Although our understanding of the molecular mechanisms that drive hair follicle morphogenesis in mice and humans has significantly increased during the last decade, still very little is known about congenital alopecias in domestic animals. Because of their rarity and the general lack of knowledge about their pathophysiology, classification of congenital alopecias in domestic animals is still unsatisfactory. This article reviews hair follicle morphogenesis and its most important molecular mechanisms, and it discusses the various forms of congenital alopecia occurring in domestic animals that have been described in the literature, differentiating between hair follicle aplasia, hair follicle dysplasia (i.e. defects associated with hair follicle development and defects associated with hair shaft formation), and neuroectodermal dysplasias, the latter involving the hair follicle pigmentary system. PMID- 17083572 TI - Evaluation of a serological test (indirect ELISA) for the diagnosis of sarcoptic mange in red foxes (Vulpes vulpes). AB - Sarcoptic mange occurs in many parts of the world and is common in populations of domestic and wild canids, including red foxes (Vulpes vulpes). In recent years, an indirect antibody enzyme-linked immunosorbent assay (ELISA), with higher sensitivity and specificity than traditional diagnostic methods, has been successfully applied in the diagnosis of sarcoptic mange in dogs. The same ELISA has also demonstrated specific antibodies to Sarcoptes scabiei in experimentally infected red foxes. The aim of this study was to evaluate the indirect ELISA when used to detect antibodies to S. scabiei in field sera from Swedish red foxes. One cohort of both infected and non-infected red foxes (cohort 1; n = 88), and one cohort of apparently non-infected foxes (cohort 2; n = 67) were examined for skin lesions and presence of S. scabiei by thorough visual examination at autopsy and skin scrapings. Samples of blood-tinted body liquid from the abdomen or thorax cavity were collected and analysed by the indirect ELISA. The relative sensitivity and specificity of the ELISA at different cut-offs (OD values) were estimated by comparing the test results to the infection status as determined by examination and skin scrapings. The highest combination of relative sensitivity and specificity, calculated based on cohort 1, was 95.4 and 100.0%, respectively. These estimates were constant for cut-offs 0.150-0.225, which included the cut off based on the mean plus three standard deviations of test results from cohort 2 (0.165). It is concluded that this test can be useful in diagnosis and epidemiological studies of S. scabiei infection in red foxes. PMID- 17083573 TI - Evaluation of the effects of topical tripeptide-copper complex and zinc oxide on open-wound healing in rabbits. AB - In this study the clinical and histopathological effects of topically applied tripeptide-copper complex (TCC) and zinc oxide on open-wound healing in rabbits was evaluated. Eighteen New Zealand rabbits were divided into three groups: TCC, zinc oxide and no treatment. One full thickness wound was created on each side of the dorsal midline in each rabbit. Wound margins were traced on days 0, 7, 14, and 21 and topical TCC and zinc oxide were applied daily for 21 days to each rabbit in the respective treatment groups. The mean unhealed wound area was significantly smaller in the TCC than in the zinc oxide group on day 7, but it was significantly smaller in the TCC group than in to the control group on days 7, 14 and 21. The mean percentage of wound contraction on day 7 was significantly higher in the TCC than in the zinc oxide group; however, it was significantly higher in the TCC group than in the control group on days 7, 14 and 21. Median time for the coverage of the wound bed with granulation tissue was significantly shorter in the TCC group than in the other groups. Filling of the open wound with granulation tissue to skin level was significantly slower in the control group than in the other two groups. Neutrophil counts decreased regardless of the group in parallel with healing, while neovascularization was best observed in the TCC group. The results suggest that TCC is a better choice in the treatment protocols of open wounds in rabbits than zinc oxide. PMID- 17083574 TI - Clinical, histological and immunohistochemical study of feline viral plaques and bowenoid in situ carcinomas. AB - Feline viral plaques (FVP) induced by papillomavirus (PV) are often hyperpigmented and flat warts. The fact that up to 47% of bowenoid in situ carcinomas (BISC), which also usually occur in the form of hyperpigmented plaques, are positive for PV antigen in immunochemistry suggests that BISC could evolve from FVP. The relationship between the presence of PV antigens and the clinical and histological features of 26 cases of feline dermatoses (clinically described as pigmented plaques and with histological diagnosis of FVP and/or BISC) was therefore determined. The cases were classified into one of the three following groups: FVP, FVP + BISC or BISC. Immunohistological detection of papillomavirus group-specific antigen was performed using a polyclonal rabbit antibovine papillomavirus antiserum. Of the seven cases in the FVP group, six were deemed positive by immunohistology as were all 10 cats in the FVP + BISC group. On the other hand, only one of the nine BISC cats was positive. The presence of both FVP and BISC lesions in some cats and the high detection rate of PV antigens in the FVP and FVP + BISC groups suggest that both conditions might have the same viral cause and that some BISC may evolve from FVP. The low rate of viral antigen detection in the BISC group indicates another cause or a loss of viral replication during the cancerogenesis. PMID- 17083575 TI - Pyoderma caused by Pseudomonas aeruginosa infection in dogs: 20 cases. AB - In this report we describe the historical, clinical, histopathological and microbiological features, as well as treatments and clinical outcome, of pyoderma where Pseudomonas aeruginosa alone was isolated on bacterial culture from lesional skin. Twenty dogs were included in this retrospective study. Seven dogs without prior history of systemic or skin disease presented with acute deep pseudomonal pyoderma characterized by a sudden onset of dorsal truncal pain. Skin lesions in these dogs consisted of erythematous papules, haemorrhagic bullae, ulcers and haemorrhagic crusts confined to the dorsum. An excellent clinical response was achieved with 3-4 weeks of treatment with oral fluoroquinolones. Thirteen dogs with a more gradual onset of skin lesions associated with pseudomonal pyoderma had a history of prior skin, ear or systemic disease and had previously been treated with antibiotics and/or immunomodulatory drugs. Skin lesions in these dogs were variable and similar to those described for superficial and deep staphylococcal pyoderma. In this group, one dog was euthanized prior to commencement of treatment, two dogs were lost to follow up, and 9 had resolution of lesions following treatment with topical silver sulfadiazine (one dog), fluoroquinolones (six dogs) or cephalexin (two dogs) administered orally for 3 to 12 weeks. Rod-shaped bacteria were not always detected on cytology. Histopathology of dogs with deep pseudomonal pyoderma was characterized by severe perforating suppurative folliculitis and furunculosis. PMID- 17083576 TI - Treatment with oral cyclosporin A of a case of vesicular cutaneous lupus erythematosus in a rough collie. AB - Cyclosporin A (CsA) is an immunosuppressive agent that can be used as alternative treatment to glucocorticoid therapy. This case report describes a case of vesicular cutaneous lupus erythematosus in a rough collie successfully controlled with oral CsA for more than 18 months. PMID- 17083577 TI - Raccoonpox in a Canadian cat. AB - Poxvirus infections affecting the skin of cats are extremely rare in North America, in contrast to Europe where cowpox virus is well recognized as an accidental pathogen in cats that hunt small rodents. The virus or viruses responsible for the anecdotal cases in North America have never been characterized. This paper reports a case of raccoonpox infection in a Canadian cat. Biopsy of the initial ulcerative lesion on the forepaw revealed ballooning degeneration of surface and follicular keratinoctyes. Infected cells contained large eosinophilic type A inclusions. Electron microscopic examination revealed virions of an orthopoxvirus, subsequently identified as raccoonpox by polymerase chain reaction and gene sequencing. The cat made a full recovery. PMID- 17083578 TI - Bacterial pseudomycetoma in dwarf hamster, Phodopus sungorus. AB - A case of a dwarf hamster with two progressively growing nodules on the right fore limb is described. These were excised following ineffective medical treatment and were submitted for histopathological examination, which revealed bacterial pseudomycetoma in both nodules. To the authors' knowledge this is the first reported case of bacterial pseudomycetoma in a dwarf hamster. PMID- 17083579 TI - Pustular dermatosis caused by fire ant (Solenopsis invicta) stings in a dog. AB - This paper reports a case of spontaneous fire ant sting dermatitis in canine. The skin lesions consisted of nonfollicular, grouped pustules in the abdominal region. Several fire ants were found attached to the lesional skin. Histopathological findings included an intraepidermal neutrophilic pustule and predominantly neutrophilic interstitial dermatitis. An extensive area of collagen degeneration was also observed below the epidermal pustule. These clinical and histopathological findings are remarkable similar to those observed in fire ant stings in humans. PMID- 17083582 TI - Sentinel lymph node biopsy in cutaneous squamous cell carcinoma: a systematic review of the English literature. AB - BACKGROUND: Although most cutaneous squamous cell carcinoma (SCC) is curable by a variety of treatment modalities, a small subset of tumors recur, metastasize, and result in death. Although risk factors for metastasis have been described, there are little data available on appropriate workup and staging of patients with high risk SCC. OBJECTIVE: We reviewed reported cases and case series of SCC in which sentinel lymph node biopsy (SLNB) was performed to determine whether further research is warranted in developing SLNB as a staging tool for patients with high risk SCC. METHODS: The English medical literature was reviewed for reports of SLNB in patients with cutaneous SCC. Data from anogenital and nonanogenital cases were collected and analyzed separately. The percentage of cases with a positive sentinel lymph node (SLN) was calculated. False negative and nondetection rates were tabulated. Rates of local recurrence, nodal and distant metastasis, and disease-specific death were reported. RESULTS: A total of 607 patients with anogenital SCC and 85 patients with nonanogenital SCC were included in the analysis. A SLN could not be identified in 3% of anogenital and 4% of nonanogenital cases. SLNB was positive in 24% of anogenital and 21% of nonanogenital patients. False-negative rates as determined by completion lymphadenectomy were 4% (8/213) and 5% (1/20), respectively. Most false-negative results were reported in studies from 2000 or earlier in which the combination of radioisotope and blue dye was not used in the SLN localization process. Complications were reported rarely and were limited to hematoma, seroma, cutaneous lymphatic fistula, wound infection, and dehiscence. CONCLUSIONS: Owing to the lack of controlled studies, it is premature to draw conclusions regarding the utility of SLNB in SCC. The available data, however, suggest that SLNB accurately diagnoses subclinical lymph node metastasis with few false-negative results and low morbidity. Controlled studies are needed to demonstrate whether early detection of subclinical nodal metastasis will lead to improved disease free or overall survival for patients with high-risk SCC. PMID- 17083583 TI - In vivo follicular unit multiplication: is it possible to harvest an unlimited donor supply? AB - BACKGROUND: Follicular unit extraction is a process of removing one follicular unit at a time from the donor region. The most important limitation of this surgical procedure is a high transection rate. OBJECTIVE: In this clinical study, we have transplanted different parts of transected hair follicle by harvesting with the follicular unit extraction technique (FUE) in five male patients. MATERIALS AND METHODS: In each patient, three boxes of 1 cm(2) are marked at both donor and recipient sites. The proximal one-third, one-half, and two-thirds of 15 hair follicles are extracted from each defined box and transplanted in recipient boxes. The density is determined at 12 months after the procedure. RESULTS: A mean of 3 (range, 2-4) of the proximal one-third, 4.4 (range, 2-6) of the proximal one-half, and 6.2 (range, 5-8) of the proximal two-thirds of the transplanted follicles were observed as fully grown after 1 year. At the donor site, the regrowth rate was a mean of 12.6 (range, 10-14) of the proximal one third, 10.2 (range, 8-13) of the proximal one-half, and 8 (range, 7-12) of the proximal two-thirds, respectively. CONCLUSION: The survival rate of the transected hair follicles is directly related to the level of transection. Even the transected parts, however, can survive at the recipient site; the growth rate is not satisfactory and they are thinner than the original follicles. We therefore recommend that the surgeon not transplant the sectioned parts and be careful with the patients whose transection rate is high during FUE procedures. PMID- 17083584 TI - Hair restoration surgery in patients with pubic atrichosis or hypotrichosis: review of technique and clinical consideration of 507 cases. AB - BACKGROUND: Pubic atrichosis or hypotrichosis is a common condition in Korean women of Mongolian origin. This results in many patients receiving hair restoration surgery, which is currently thought to be the only definitive therapy. OBJECTIVE: To pursue more natural and realistic-appearing results, to define patient characteristics, and to estimate the survival rate of transplanted pubic hair through restoration surgery, we examined our cases of pubic hair restoration surgery with single-hair grafts. METHODS: We selected 507 patients with pubic atrichosis or hypotrichosis who visited for pubic hair transplantation between March 1, 2001, and February 28, 2005. We reviewed the medical charts of the 507 patients and performed statistical analysis. We also carried out a detailed evaluation of our surgical technique to 100 patients. In addition, 20 patients, who agreed to participate in the study for survival rate, had received transplantation of 40 hairs in a 1.5 x 1.5-cm area after the angular points were tattooed. The number of hairs grown after 1 year of transplantation was counted in each case. RESULTS: Among the 507 subjects, 169(33.3%) were in their 40s. The mean (+/-SD) patient age was 41.3+/-10.8 years. Of these, 115 patients (22.7%) had pubic atrichosis, and 392 patients (77.3%) had pubic hypotrichosis. In addition, 81.7% of atrichosis patients had a family history of atrichosis or hypotrichosis. Pubic atrichosis accompanied axillary atrichosis or hypotrichosis in 60.0 and 38.2% of the cases, respectively. The most common reason for the hair restorative procedure was the subject's sense of inferiority to the same sex (73.8%). The mean number of transplanted hairs was 929.3+/-76.6. The most common design pattern that we used was modified horizontal type (87.0%). The mean survival rate of single-hair grafts on the pubis was 73.6+/-7.6%. CONCLUSION: This study suggested that pubic hair transplantation surgery is a suitable cosmetic procedure to address the inferiority complex of patients with pubic atrichosis or hypotrichosis. Knowledge of natural pubic pattern and normal physiologic features is essential to create a natural and realistic appearance in a given subject. PMID- 17083585 TI - Safety and efficacy of poly-L-lactic acid injections in persons with HIV associated lipoatrophy: the US experience. AB - BACKGROUND: With the introduction of highly active antiretroviral therapy (HAART), HIV/AIDS has become a more chronic disease. This increase in survival, however, has been accompanied by the occurrence of metabolic and morphologic changes known collectively as HIV-associated lipodystrophy. OBJECTIVE: The objective was to evaluate the quantifiable improvement in facial wasting (lipoatrophy) after serial injections of poly-L-lactic acid (PLLA) as well as the long-term safety and durability of this material. METHODS AND MATERIALS: A single site, open-label study in which patients received up to six treatment sessions of injectable PLLA. Patients were followed for 12 months after treatment. Patients were evaluated for increase in total cutaneous thickness by skin calipers. Serial photographs and patient satisfaction and patient well-being questionnaires were undertaken throughout the study period. RESULTS: Patients at the end of treatment exhibited a mean increase in skin thickness of 65.1% compared with baseline values. This correction was maintained throughout the 12-month follow-up period (68.8% at 6 months and 73% at 12 months). On a one to five scale, patient satisfaction was 4.5 at the end of treatment and increased to 4.8 at 12-month follow-up. No serious adverse events were reported. CONCLUSION: PLLA is a safe and well-tolerated treatment option for HIV-associated lipoatrophy. PMID- 17083586 TI - Posterolateral thigh perforator varicosities in 12 patients: a normal deep venous system and successful treatment with ultrasound-guided sclerotherapy. AB - OBJECTIVES: To determine whether in patients with posterolateral thigh perforator (PLTP) varicosities ultrasound-guided sclerotherapy (USGS) is successful and to determine any deep venous anomaly. PATIENTS: Twelve consecutive patients with PLTP varicosities were investigated and treated. DESIGN: In this prospective intervention study USGS of the PLTP with polidocanol microfoam 1-2% was performed with a maximal number of three treatments. Clinical pictures and Duplex imaging were performed before and after treatment. Ascending phlebography was performed in 9 patients. RESULTS: Ten patients showed a marked success, defined as no PLTP varicosities visible and no reflux detectable. In 2 patients there was moderate success. The mean number of treatments was 1.58, the mean diameter of the PLTP vein was reduced from 4.06 (range, 2.2-6.1) to 1.97 (range, 0-3.3) mm (p=0.003, Wilcoxon signed rank test). No serious side effects were noted. No deep venous anomaly was found in the phlebographic study. CONCLUSION: In patients with PLTP varicosities, USGS is very successful, with very few side effects. No deep venous anomalies were found in our study, which conforms to the literature. PMID- 17083587 TI - Approaches to document the efficacy and safety of microdermabrasion procedure. AB - BACKGROUND: Microdermabrasion is a popular cosmetic procedure for skin rejuvenation, which is achieved by mechanical abrasion of the skin at a superficial level. OBJECTIVE: The objective was to study the efficacy of microdermabrasion in photoaging and to investigate the compatibility of a cleanser and a lotion with microdermabrasion. METHODS: Sixteen women underwent microdermabrasion to the face once a week for a total of six treatments. Subjects were also given a personal skin care regimen (cleanser and lotion). Colorimetry values as well as investigator and patients ratings for safety and efficacy were analyzed at various time points and compared to baseline. RESULTS: There were no significant changes in safety and tolerance variables throughout the study. Clinical efficacy variables (fine wrinkles, dullness, pigmentation, and large pores) significantly improved by the third treatment, with further improvement by the end of the study (six treatments). Subjects perceived improvement in facial photoaging variables. Colorimetry showed increased brightness and decreased yellowness of target skin sites on the face throughout the study. CONCLUSION: Multiple microdermabrasions were effective in significantly improving various facial photoaging variables. The personal skin care regimen used was well tolerated by the subjects. PMID- 17083588 TI - Evaluation of natural change of skin function in split-thickness skin grafts by noninvasive bioengineering methods. AB - BACKGROUND: Autologous split-thickness skin grafts (STSGs) are considered the mainstay for the treatment of large full-thickness wounds. There have been few studies reporting the natural change of the skin function in STSGs after procedure, however. OBJECTIVE: The objective was to evaluate the natural change of the skin function in STSG using noninvasive bioengineering methods. METHODS: Eighteen patients were eligible for the study. The skin functions of the graft and the control site were evaluated by an evaporimeter, corneometer, mexameter, and cutometer at Postoperation Days 0.5, 1, 2, 3, 6, 9, and 12 months. RESULTS: Transepidermal water loss (TEWL) of the graft was maintained around that of the normal skin. The values of the skin hydration testing generally decreased during the follow-up period. Erythema was highly maintained for the whole period. For the pigmentation, the ratio tended to increase after 6 months. The skin pliability of the graft was abruptly decreased at 0.5 month, and it recovered from 3 to 12 months. The value did not reach that of the normal skin, however. CONCLUSION: Our results showed that the STSGs had changed within the frame of the skin function, including the TEWL, epidermal hydration, color, and pliability, throughout 1 year after surgery. The authors have indicated no significant interest with commercial supporters. PMID- 17083589 TI - The Australian Mohs database: short-term recipient-site complications in full thickness skin grafts. AB - BACKGROUND: There are only a few reports on the outcome with full-thickness skin grafts (FTSG) in defect reconstruction after Mohs micrographic surgery (MMS). OBJECTIVES: The objective was to evaluate the complications with FTSG in patients treated with MMS for cutaneous tumors. METHODS: This study is based on a prospective data collection of all patients in Australia treated with MMS and monitored by the Skin and Cancer Foundation, between 1993 and 2002. Primary outcome measures were FTSG recipient-site complications (partial/complete graft failure, graft infection, acute bleeding/hematoma, graft hypertrophy, and graft contracture). RESULTS: The study group included 2,673 patients (50.2% men) with a mean age of 64+/-14 years. Most tumors were basal call carcinoma (90.9%) and were located on the nose (63.8%), periocular area (14.8%), or auricular area (8.1%). A total of 11.7% of cases had recipient-site complications (graft hypertrophy, 42.3%; partial graft failure, 27.2%; graft contraction, 15.3%). Only 45.2% of complicated cases required treatment. CONCLUSION: Short-term recipient-site complications occurred in less than 12% of reconstructions with FTSG. Graft hypertrophy and partial graft failure accounted for most cases of complications. The authors have indicated no significant interest with commercial supporters. PMID- 17083590 TI - Scalp tumors treated with Mohs micrographic surgery: clinical features and surgical outcome. AB - BACKGROUND: There are only a small number of reports on the outcome of scalp tumors treated with Mohs micrographic surgery (MMS). OBJECTIVE: The objective was to present a large series of patients with scalp tumors treated with MMS in Australia between 1993 and 2002. METHODS: This prospective, multicenter study included all patients with scalp tumors who were monitored by the Skin and Cancer Foundation Australia. The variables analyzed were patient demographics, reason for referral, preoperative tumor size and postoperative defect size, recurrences before MMS, histologic subtypes, perineural invasion, and 5-year recurrence. RESULTS: The study included 316 patients (68% men) with a mean age of 65+/-15 years. The most common tumors were basal cell carcinoma (BCC), 57.9%; squamous cell carcinoma (SCC), 35.8%; Bowen's disease (BD), 4.1%; and atypical fibroxanthoma (AFX), 1.6%. Recurrent tumors comprised 37% of cases. The recurrence rate for 70 BCC patients who completed the 5-year follow-up was 5.7%, and for 31 SCC cases it was 3.2%. No cases of recurrence were noted in the patients with BD and AFX. CONCLUSION: BCC was the most common scalp tumor managed by MMS. The low 5-year recurrence rate emphasizes the importance of margin controlled excision of scalp tumors. PMID- 17083591 TI - Difference between pigmented and nonpigmented basal cell carcinoma treated with Mohs micrographic surgery. AB - BACKGROUND: There have been several articles characterizing cases of pigmented basal cell carcinomas (PBCC). Previous studies have also evaluated the relationship between histologic pattern and frequency of basal cell carcinoma (BCC) associated with pigment formation. No specific studies, however, have examined the subclinical extension and surgical margins of PBCC tumors. OBJECTIVE: A prospective study of 345 Mohs micrographic BCC surgical cases revealed 67 PBCC cases. Analysis of patient details included patient age and sex, the lesional site, histologic subtype, tumor size, final surgical margin, and the number of stages required to achieve tumor-free margins together with the presence or absence of pigment. METHODS: This study was performed between May 2004 and January 2005 at the Department of Dermatology and Cutaneous Surgery, University of Miami, Mohs Surgery Center. RESULTS: Total mean surgical margin was smaller in the PBCC than the nonpigmented BCC (NPBCC) group (3.89 mm vs. 5.85 mm; p<0.05). In lesions less than 2 cm in size, there were even more significant differences between the two groups (3.32 mm vs. 5.33 mm; p<0.05), and also between the aggressive and nonaggressive histologically diagnosed groups (3.13 mm vs. 5.01 mm; p<0.05). CONCLUSIONS: We have demonstrated that PBCC requires a smaller surgical margin for complete tumor excision than NPBCC, especially in smaller tumors and in the nonaggressive histologic subtype group. Treatment within the early growth stages also involves less subclinical microscopic invasion and a smaller surgical margin in PBCC. PMID- 17083592 TI - Photorejuvenation by intense pulsed light with objective measurement of skin color in Japanese patients. AB - BACKGROUND AND OBJECTIVES: This study had two objectives: subjective evaluation of overall skin rejuvenation effects of relatively short-wavelength intense pulsed light (IPL) and objective changes in basic skin tone as measured by a spectrophotometer. STUDY DESIGN/MATERIALS AND METHODS: Twenty-five women selected at random received a series of three IPL treatments. Efficacy was evaluated over a 3-month follow-up period. Concurrently, a spectrophotometer was used to measure "lightness" (L(*)) to quantify the lightening effect changes to pretreatment and posttreatment basic skin tone. RESULTS: Subjective improvement of 50% or more was seen in 18 of 25 patients for pigmentation. One patient showed exacerbation of latent epidermal melasma as a complication. In the spectrophotometric analysis, the mean value of L(*) increased from a baseline value of 60.86 to 63.22, at 3 month follow-up period, with statistical significance. CONCLUSION: IPL skin rejuvenation using relatively shorter wavelengths and pulse widths brought about significant macroscopic and quantitative improvements, especially in the treatment of epidermal pigmentation and improvement of basic skin tone. PMID- 17083593 TI - A simple, pain-free treatment for ingrown toenails complicated with granulation tissue. AB - BACKGROUND: The treatment of ingrown toenail complicated with granulation tissue is usually partial or total nail avulsion with or without matricectomy. It costs loss of occupational power, however, because most patients cannot go to work or school for some time after surgery, and it is a costly and uncomfortable procedure for most patients. OBJECTIVE: This study aimed to find an easy, painless, and inexpensive alternative. MATERIALS AND METHODS: Seven patients with ingrown toenails complicated with granulation tissue are included. A small apparatus was applied on the nails, granulation tissue was chemically cauterized, and a foot bath was recommended twice daily. They were followed on a weekly basis or every other week until recovery. None of the patients received systemic treatment. RESULTS: All seven patients were completely cured without requiring surgery and/or systemic treatment. The procedure did not have any effect on their daily life. The follow-up examination of the patients at 6 months revealed that they were totally cured, and there were no recurrences. CONCLUSION: Patients with ingrown toenails complicated by granulation tissue might have an inexpensive and pain-free treatment alternative, although new studies with more patients are required. PMID- 17083594 TI - Hypoepiluminescence microscopy of pigmented skin lesions: new approach to improve recognition of dermoscopic structures. AB - BACKGROUND: Hypoepiluminescence microscopy (HELM) is a new dermoscopic approach for analysis of pigmented skin lesions (PSLs) after surgical excision. OBJECTIVES: The objective was to verify whether this method could provide additional morphologic information for diagnostic or didactic purposes compared to conventional epiluminescence microscopy (ELM). PATIENTS AND METHODS: Thirty one PSLs from 30 patients were consecutively evaluated by ELM and, after excision, by HELM. For HELM examination, the lesion was positioned on a glass slide and illuminated from above with a halogen lamp and from underneath with an LED source. All lesions were subsequently examined histopathologically. RESULTS: In 11 of 31 (35.5%) lesions, a typical pigment network, as assessed by ELM, appeared bidimensional with HELM. In 9 lesions (9/31; 29%) ELM showed a gray-blue area, while HELM allowed us to distinguish 5 lesions (5/9, 55.5%) with gray area predominant showing a lichenoid lymphocytic infiltration and few melanophages from the other 4 lesions (4/9, 44,5%) with heavy dermal accumulation of pigmented melanocytes or melanophages where a blue area was clearly visible at HELM. In 9 other cases (29%), ELM analysis revealed a central homogeneous dark brown/black pigmentation that in 7 cases was seen under HELM examination to consist of globules. CONCLUSIONS: HELM is particularly useful in evaluating heavily PSLs or structures located in the reticular dermis. PMID- 17083595 TI - Nonmelanocytic lesions defying the two-step dermoscopy algorithm. AB - The first step of the two-step algorithm of dermoscopy aims at differentiating melanocytic from nonmelanocytic pigmented lesions, using a stepwise evaluation for the presence of specific dermoscopic criteria. The purpose of this article is to heighten awareness of clinicians to nonmelanocytic lesions that defy the two step algorithm, thus simulating melanocytic lesions dermoscopically. Seborrheic keratosis, solar lentigo, dermatofibroma, and supernumerary accessory nipple may present with network-like structures. Seborrheic keratosis, dermatofibroma, subcorneal hemorrhage, basal cell carcinoma (BCC), and cutaneous metastases of breast and other cancers may contain pigmented globules. Peripheral streaks can also be seen in seborrheic keratosis and BCC. Homogenous bluish pigmentation, simulating a blue nevus, can also be seen in benign vascular lesions, Kaposi sarcoma, radiation tattoo, and BCC. This overlap of features between melanocytic and nonmelanocytic lesions suggests that integration of all dermoscopic features in the lesion, rather than a stepwise evaluation, may facilitate reaching the correct diagnosis in select cases as outlined in this article. PMID- 17083596 TI - Persistent granulomatous inflammatory response induced by injectable poly-L lactic acid for HIV lipoatrophy. PMID- 17083597 TI - Migraine ophthalmique with reversible scotomas after sclerotherapy with liquid 1% polidocanol. PMID- 17083598 TI - Long-pulsed dye laser for the treatment of erythromelanosis follicularis faciei: report of two clinical cases. PMID- 17083599 TI - Rapid growth of basal cell carcinoma in a multigestational pregnancy. PMID- 17083600 TI - Repair of basal cell carcinoma on the outer canthus via a bifurcated island flap. PMID- 17083601 TI - Indirect electrosurgery. PMID- 17083602 TI - Suture marks: factors of causation and prevention. PMID- 17083603 TI - Iatrogenic nasolabial asymmetry. PMID- 17083605 TI - Vive la difference. PMID- 17083604 TI - Proteasomal chymotrypsin-like peptidase activity is required for essential functions of human monocyte-derived dendritic cells. AB - The ubiquitin-proteasome pathway is the principal system for extralysosomal protein degradation in eukaryotic cells, and is essential for the regulation and maintenance of basic cellular processes, including differentiation, proliferation, cell cycling, gene transcription and apoptosis. The 26S proteasome, a large multicatalytic protease complex, constitutes the system's proteolytic core machinery that exhibits different proteolytic activities residing in defined proteasomal subunits. We have identified proteasome inhibitors - bortezomib, epoxomicin and lactacystin - which selectively inhibit the proteasomal beta5 subunit-located chymotrypsin-like peptidase activity in human monocyte-derived dendritic cells (DCs). Inhibition of proteasomal chymotrypsin-like peptidase activity in immature and mature DCs impairs the cell surface expression of CD40, CD86, CD80, human leucocyte antigen (HLA)-DR, CD206 and CD209, induces apoptosis, and impairs maturation of DCs, as demonstrated by decreased cell-surface expression of CD83 and lack of nuclear translocation of RelA and RelB. Inhibition of chymotrypsin-like peptidase activity abrogates macropinocytosis and receptor-mediated endocytosis of macromolecular antigens in immature DCs, and inhibits the synthesis of interleukin (IL)-12p70 and IL-12p40 in mature DCs. As a functional consequence, DCs fail to stimulate allogeneic CD4(+) and CD8(+) T cells and autologous CD4(+) T cells sufficiently in response to inhibition of chymotrypsin-like peptidase activity. Thus, proteasomal chymotrypsin-like peptidase activity is required for essential functions of human DCs, and inhibition of proteasomal chymotrypsin-like peptidase activity by selective inhibitors, or by targeting beta5 subunit expression, may provide a novel therapeutic strategy for suppression of deregulated and unwanted immune responses. PMID- 17083606 TI - The effect of nitrous oxide on halothane, isoflurane and sevoflurane requirements in ventilated dogs undergoing ovariohysterectomy. AB - OBJECTIVE: To examine the effect of 64% nitrous oxide (N2O) on halothane (HAL), isoflurane (ISO) or sevoflurane (SEV) requirements in dogs undergoing ovariohysterectomy. STUDY DESIGN: Prospective, randomized, clinical trial. ANIMALS: Ninety, healthy dogs of (mean +/- SD) body weight 21.2 +/- 10.0 kg and age 17.8 +/- 22.8 months. MATERIALS AND METHODS: After premedication with acepromazine, hydromorphone and glycopyrrolate, anesthesia was induced with thiopental administered to effect. Dogs received one of six inhalant protocols (n = 15 group): HAL; HAL/N2O; ISO; ISO/N2O; SEV; or SEV/N2O. End-tidal CO2 was maintained at 40 +/- 2 mmHg with intermittent positive pressure ventilation (IPPV). Body temperature, heart rate, indirect systemic arterial blood pressures, inspired and end-tidal CO2, volatile agent, N2O and O2 were recorded every 5 minutes. The vaporizer setting was decreased in 0.25-0.5% decrements to elicit a palpebral reflex, and this level maintained. Statistical analysis included two way anova for repeated measures with Bonferroni's correction factor and statistical significance assumed when p < 0.05. Percentage reduction in end-tidal volatile agent was calculated at 60 minutes after starting study. RESULTS: End tidal HAL, ISO and SEV decreased when N2O was administered. Percentage reduction: HAL (12.4%); ISO (37.1%) and SEV (21.4%). Diastolic, mean and systolic blood pressures increased in ISO/N2O compared with ISO. Heart rate increased in ISO/N2O and SEV/N2O compared with ISO and SEV, respectively. Systolic, mean and diastolic blood pressures increased in SEV compared with HAL and ISO. Systolic, mean, diastolic blood pressures and heart rate increased in SEV/N2O and ISO/N2O compared with HAL/N2O. CONCLUSIONS: N2O reduces HAL, ISO and SEV requirements in dogs undergoing ovariohysterectomy. Cardiovascular stimulation occurred when N2O was used with ISO, less so with SEV and not with HAL PMID- 17083607 TI - Minimum alveolar concentration of desflurane in llamas and alpacas. AB - OBJECTIVE: To determine the minimum alveolar concentration (MAC) of desflurane in llamas and alpacas. DESIGN: Prospective study. Animals Six healthy adult llamas and six healthy adult alpacas. PROCEDURE: Anesthesia was induced with desflurane delivered with oxygen through a mask. An endotracheal tube was inserted, and a port for continuous measurement of end-tidal and inspired desflurane concentrations was placed between the endotracheal tube and the breathing circuit. After equilibration at an end-tidal-to-inspired desflurane concentration ratio >0.90 for 15 minutes, a 50-Hz, 80-mA electrical stimulus was applied to the antebrachium until a response was obtained (i.e. gross purposeful movement) or for up to 1 minute. The vaporizer setting was increased or decreased to effect a 10-20% change in end-tidal desflurane concentration, and equilibration and stimulus were repeated. The MAC was defined as the average of the lowest end tidal desflurane concentration that prevented a positive response and the highest concentration that allowed a positive response. RESULTS: Mean +/- SD MAC of desflurane was 7.99 +/- 0.58% in llamas and 7.83 +/- 0.51% in alpacas. CONCLUSIONS AND CLINICAL RELEVANCE: The MAC of desflurane in llamas and alpacas was in the range of that reported for other species. PMID- 17083608 TI - Differences in need for hemodynamic support in horses anesthetized with sevoflurane as compared to isoflurane. AB - OBJECTIVE: To study whether hemodynamic function in horses, particularly mean arterial blood pressure (MAP), is better maintained with sevoflurane than isoflurane, thus requiring less pharmacological support. STUDY DESIGN: Prospective randomized clinical investigation. Animals Thirty-nine racehorses undergoing arthroscopy in lateral recumbency. METHODS: Horses were assigned to receive either isoflurane (n = 20) or sevoflurane (n = 19) at 0.9-1.0 minimum alveolar concentration (MAC) for maintenance of anesthesia. Besides routine clinical monitoring, cardiac output (CO) was measured by lithium dilution. Hemodynamic support was prescribed as follows: when MAP decreased to <70 mmHg, patients were to receive infusion of 0.1% dobutamine, which was to be discontinued at MAP >85 mmHg or heart rate >60 beats minute(-1). Statistical analysis of results, given as mean +/- SD, included a clustered regression approach. RESULTS: Average inhalant anesthetic time [91 +/- 35 (isoflurane group) versus 97 +/- 26 minutes (sevoflurane group)] and dose (in MAC multiples), volume of crystalloid solution infused, and cardiopulmonary parameters including CO were similar in the two groups, except heart rate was 8% higher in isoflurane than sevoflurane horses (p < 0.05). To maintain MAP >70 mmHg, isoflurane horses received dobutamine over a significantly longer period (55 +/- 26 versus 28 +/- 21% of total anesthetic time, p < 0.01) and at a 51% higher dose than sevoflurane horses (41 +/- 19 versus 27 +/- 23 microg kg(-1) MAC hour(-1); p = 0.058), with 14/20 isoflurane animals and only 9/19 sevoflurane horses being infused with dobutamine at >30 microg kg(-1) MAC hour(-1) (p < 0.05). Dobutamine infusion rates were consistently lower in the sevoflurane as compared to the isoflurane group, with differences reaching significance level during the 0-30 minutes (p < 0.01) and 61-90 minutes periods (p < 0.05). CONCLUSIONS AND CLINICAL RELEVANCE: Horses under sevoflurane anesthesia may require less pharmacological support in the form of dobutamine than isoflurane-anesthetized horses. This could be due to less suppression of vasomotor tone. PMID- 17083609 TI - Effects of isovolemic resuscitation with hemoglobin-based oxygen carrier Hemoglobin glutamer-200 (bovine) on systemic and mesenteric perfusion and oxygenation in a canine model of hemorrhagic shock: a comparison with 6% hetastarch solution and shed blood. AB - OBJECTIVE: To study Hemoglobin glutamer-200 bovine (Hb-200), 6% hetastarch (HES) and shed whole blood (WB) resuscitation in canine hemorrhagic shock. STUDY DESIGN: Prospective laboratory investigation. Animals Twelve adult dogs [29 +/- 1 kg (mean +/- SD)]. METHODS: Anesthetized dogs were instrumented for recording systemic and mesenteric hemodynamic parameters and withdrawal of arterial, mixed and mesenteric venous blood, in which hematological, oxygenation, blood gas and acid-bases variables were determined. Recordings were made before [baseline (BL)], after 1 hour of hypovolemia and immediately and 3 hours post-resuscitation with 30 mL kg(-1) of either Hb-200, HES, or WB. RESULTS: Blood withdrawal (average 34 +/- 2 mL kg(-1)) caused significant hemodynamic changes, metabolic acidosis and hyperlactatemia characteristic for hemorrhagic shock. Only WB transfusion restored all variables. Hemoglobin glutamer-200 bovine infusion returned most hemodynamic parameters including cardiac output and mesenteric arterial blood flow to BL but increased mean arterial pressure above BL (p < 0.05). However, Hb-200 failed to restore total Hb and arterial oxygen content (CaO2), leaving systemic (DO2I) and mesenteric O2 delivery (DO2Im) below BL (p < 0.05). Nevertheless, acid-base variables recovered completely after Hb-200 resuscitation, and met-hemoglobin (Met-Hb) levels increased (p < 0.05). Hetastarch resuscitation returned hemodynamic variables to or above BL but further decreased total Hb and CaO2, preventing recovery of sDO2I and mDO2I (p < 0.05). Thus, systemic and mesenteric O2 extraction stayed above BL (p < 0.05) while acid-base variables recovered to BL, although slower than in Hb-200 and WB groups (p < 0.05). CONCLUSIONS AND CLINICAL RELEVANCE: Resuscitation with Hb-200 seemed to resolve metabolic acidosis and lactatemia more rapidly than HES, but not WB; yet it is not superior to HES in improving DO2I and DO2Im. The hyperoncotic property of solutions like Hb-200 that results in rapid volume expansion with more homogenous microvascular perfusion and the ability to facilitate diffusive O2 transfer accelerating metabolic recovery may be the key mechanisms underlying their beneficial effects as resuscitants. PMID- 17083610 TI - Postanesthetic hyperthermia in cats: a retrospective comparison between hydromorphone and buprenorphine. AB - OBJECTIVE: To determine the prevalence of postanesthetic hyperthermia [rectal temperature >40 degrees C (104 degrees F)] in a clinical population of cats. STUDY DESIGN: Retrospective study. ANIMAL POPULATION: One hundred and twenty-five cats with an age range of 2 months to 16.1 years, and weighing 3.9 +/- 1.5 kg. MATERIALS AND METHODS: Data were obtained from the medical records of 125 cats that underwent general anesthesia. Information on perioperative rectal temperatures, breed, sex, weight, surgical procedure, anesthetic time, surgery time, anesthetic and analgesic drugs were retrieved. STATISTICAL ANALYSES: Five groups of cats were compared; group 1 (n = 15) received acepromazine and no opioids; group 2 (n = 17) received acepromazine and buprenorphine; group 3 (n = 19) received acepromazine, buprenorphine and ketoprofen; group 4 (n = 45) received acepromazine and hydromorphone and group 5 (n = 29) received acepromazine, hydromorphone and ketoprofen. Data conformed to a split-plot repeated measures analysis of variance and was analyzed using SAS PROC MIXED. Post hoc tests were by means of Bonferroni t-test; < or = 0.05 was considered significant. RESULTS: Rectal temperature was significantly decreased in all groups at the end of anesthesia. Rectal temperature was significantly elevated at 1, 1.5, 2, 3, 4 and 5 hours after the end of anesthesia in group 4, and at 2, 3 and 4 hours in group 5. Sixty-four percent of cats in group 4 and 69% in group 5 had rectal temperatures >40 degrees C (104 degrees F) at one or more times in the postanesthetic period. The highest temperature recorded was 42.5 degrees C (108.5 degrees F) in one cat in group 4. Mean rectal temperature did not exceed the preoperative temperature at any time during the postanesthetic period in group 1, 2 and 3 animals. CONCLUSIONS: This study indicates an association between hyperthermia and perioperative administration of hydromorphone in cats. CLINICAL RELEVANCE: When hydromorphone is used in cats their body temperature should be closely monitored. PMID- 17083611 TI - A comparison of four methods of analgesia in cats following ovariohysterectomy. AB - OBJECTIVE: To evaluate the effectiveness of preoperative administration of oral carprofen, subcutaneous ketoprofen, and local nerve block with bupivacaine in preventing postoperative pain-associated behavior in cats after ovariohysterectomy. ANIMALS: Fifty-two female intact cats. Materials and methods Cats received butorphanol (0.44 mg kg(-1) IM), carprofen (2.2 mg kg(-1) PO), ketoprofen (2.2 mg kg(-1) SQ), or bupivacaine infiltration block (1.1 mg kg(-1) SQ) before surgery. Cortisol and drug concentrations and visual analog scale (VAS) and interactive visual analog scale (IVAS) pain-associated behavior scores were measured 2 hours before and 0, 1, 2, 4, 8, 12, and 24 hours after ovariohysterectomy. RESULTS: Cats receiving butorphanol had significantly increased IVAS scores 2 hours after surgery compared with baseline measurements. Cats receiving carprofen, ketoprofen, and bupivacaine had significant increases from baseline in VAS and IVAS scores 1 and 2 hours after surgery. VAS and IVAS scores for cats receiving bupivacaine were significantly greater 1 and 2 hours after surgery than for cats that received butorphanol. Cats receiving carprofen had significant increases in cortisol 1 hour after surgery and significant decreases 24 hours after surgery compared with baseline measurements. CONCLUSIONS AND CLINICAL RELEVANCE: Preoperative carprofen and ketoprofen have effects on pain-associated behavior similar to butorphanol in cats undergoing ovariohysterectomy. Cats receiving bupivacaine blocks may require additional analgesics immediately after surgery. PMID- 17083612 TI - Investigation of the interaction between buprenorphine and sufentanil during anaesthesia for ovariectomy in dogs. AB - OBJECTIVE: To investigate the effect of buprenorphine pre-treatment on sufentanil requirements in female dogs undergoing ovariectomy. STUDY DESIGN: Randomized, 'blinded', prospective clinical study. ANIMALS: Thirty healthy female dogs referred for ovariectomy. MATERIALS AND METHODS: Dogs were randomly assigned to one of two pre-anaesthetic treatment groups. Those in the buprenorphine group (B) received buprenorphine 20 microg kg(-1) and acepromazine 0.03 mg kg(-1) IM. Control group (C) animals received an equal volume of NaCl 0.9% and acepromazine 0.03 mg kg(-1) IM. The anaesthetic technique was identical in both groups. Pre anaesthetic medication consisted of intravenous (IV) sufentanil (1.0 microg kg( 1)) and midazolam (0.05 mg kg(-1)) and intramuscular atropine (0.03 mg kg(-1)). Anaesthesia was induced with propofol and maintained with a constant rate infusion of sufentanil (1.0 microg kg(-1) hour(-1)) and with oxygen-isoflurane. Ventilation was controlled mechanically. Ovariectomy was performed using a standard technique. Baseline heart rate (HR) and direct mean arterial blood pressure (MAP) were recorded before the first incision. Increases in HR and MAP of > or =20% over baseline and, or spontaneous ventilation were controlled using IV sufentanil (1.0 microg kg(-1)) repeated after 5 minutes if haemodynamic variables remained elevated or attempts at spontaneous ventilation persisted. Analysis of variance was used to determine group differences in mean and median HR and MAP and to compare the maximum HR and MAP attained during surgery. Poisson regression was used to compare the number of sufentanil injections required in both groups. RESULTS: Group B required 2.46 times more sufentanil injections (p = 0.00487) than dogs in group C to maintain haemodynamic stability and prevent spontaneous ventilation during surgery. Group B dogs also had a significantly higher (p = 0.034) marginal mean of the log maximum MAP (4.756 +/- 0.036) compared with group C (4.642 +/- 0.036). CONCLUSIONS: Pre-treatment with buprenorphine appears to negatively influence the antinociceptive efficacy of intra-operative sufentanil. CLINICAL RELEVANCE: Withholding buprenorphine therapy 6-8 hours before anaesthesia incorporating pure mu receptor agonists is probably advisable. Alternative methods of analgesia should be provided in this period. PMID- 17083613 TI - Is the activity of partially agonistic MHC:peptide ligands dependent on the quality of immunological help? AB - CD8(+) cytotoxic T lymphocytes (CTL) are important for the immunological control of infections and tumours. Engagement of the T-cell receptor (TCR) with major histocompatibility complex (MHC) class I/peptide complexes on antigen-presenting cells (APC) is the key interaction, which initiates the process of T-cell activation. Depending on the affinity of this interaction, different arrays of signalling pathways and functional outcomes can be activated in the specific T cells. Molecular alterations in the peptide bound to the MHC class I can lead to a lower affinity of the MHC:TCR interaction resulting in incomplete or qualitatively different T-cell responses. Altered peptide ligands (APL) exhibiting such activity are referred to as partial agonists and often occur naturally through genetic instability, which affects T-cell epitopes derived from rapidly mutating viruses or tumour-associated cellular antigens. Partial agonists are usually viewed as peptide variants, which escape efficient CTL recognition. Our recent data suggest that APL can not only trigger incomplete activation but also induce and modulate intrinsic T-cell programmes leading to the shut-off of specific CTL responses. This APL-induced suppression appears to be more prominent in the absence of immunological help, suggesting that under conditions of immune deregulation APL may actively inhibit CTL responses against infectious agents or tumours. In this review, we discuss experimental data supporting this model and possible role of APL-induced immunosuppression in different pathological conditions. PMID- 17083614 TI - Effects of RNA interference on CD80 and CD86 expression in bone marrow-derived murine dendritic cells. AB - To investigate whether RNA interference (RNAi) induced by small interfering RNA (siRNA) could suppress CD80 and CD86 expression in bone marrow-derived murine dendritic cells (DC). The bone marrow-derived DC of mice were separated and cultured in vitro, chemically synthesized siRNA were then transferred into the cells by LipofectAMINE 2000, and the siRNA transfection efficacy was assessed by both fluorescence microscope and flow cytometry. The mRNA expression and protein synthesis were analysed by real-time RT-PCR and flow cytometry. The cell viability of transfected DC was determined by annexin V and propidium iodine staining. Transfection of bone marrow-derived murine DC with a non-silencing FITC labelled control siRNA demonstrated a high (71.86%) transfection efficiency without affecting cellular viability. CD80-1 siRNA was the most effective siRNA to block CD80 expression in three candidates. Similarly, CD86-3 siRNA was extraordinarily effective in repressing the expression of CD86. Cotransfection of siRNA specific to CD80 and CD86 can enhance gene silencing that is not affected by DC activation-inducing signals. CD80 and CD86 siRNA suppressed the expression of CD80 and CD86 to 31.05 +/- 2.41% and 25.43 +/- 0.85%, respectively, of the level in untreated cells (P < 0.05). siRNA is capable of triggering RNAi in bone marrow-derived DC; it can specifically and effectively knock down CD80 and CD86 gene expression. This approach is a useful tool by which costimulatory molecules of DC can be studied as well as a potential therapeutic option for allograft rejection. PMID- 17083615 TI - Absence of CD4+ T cells impairs host defence of mice infected with Fonsecaea pedrosoi. AB - Chromoblastomycosis is a human chronic, often debilitating, suppurative, granulomatus mycosis of the skin and subcutaneous tissues beginning after inoculation trauma. It occurs worldwide, but is more frequently observed in tropical countries such as Brazil. Some studies have focused on fungus-host interaction, showing a predominantly cell-mediated immune response, with the activation of macrophages involved in fungus phagocytosis. Immunization with live conidia produced a high influx of CD4 T cells into the draining lymph node. The sensitized T cells proliferate in vitro when restimulated with specific antigen and preferentially produce IFN- gamma. To better characterize the role played by T cells on the chromoblastomycosis infection we used mice deficient for CD4 and CD8. Data determined by CFU counts associated with decreased DTH and IFN-gamma production of infected mice clearly demonstrated that, during experimental F. pedrosoi infection, absence of CD4(+) cells induces a more severe disease. PMID- 17083616 TI - Long-term haematopoietic reconstitution and survival without interleukin-7 in a murine syngeneic bone marrow transplantation model. AB - We created a syngeneic mouse bone marrow transplantation (BMT) model to examine the effect of endogenous interleukin-7 (IL-7) on long-term (>or=140 days) haematopoietic reconstitution and survival after BMT. Wild-type (WT) IL-7(+/+) and knockout (KO) IL-7(-/-) mice were lethally irradiated and transplanted with bone marrow. Survival is best (85.7%) in the group WT grafts transplanted into WT recipients (WT-->WT) with a trend towards poorer survival in the other groups (WT ->KO: 60%, KO-->WT: 50%, KO-->KO: 45.5%, differences statistically not significant). If the recipient is deficient for IL-7-producing cells, T- and B cell reconstitution remain incomplete. If the graft lacks IL-7-producing cells there is a significant delay in T- and NK-cell reconstitution. Interestingly, in the absence of IL-7, T-cell reconstitution is neither delayed nor incomplete because of an expansion of TCRalphabeta(+)/CD4(-)/CD8(-) double negative T cells. Long-term survival and lymphocyte reconstitution after syngeneic BMT can occur despite the absence of IL-7. PMID- 17083617 TI - Decreased expression of VEGF-A in rat experimental autoimmune encephalomyelitis and in cerebrospinal fluid mononuclear cells from patients with multiple sclerosis. AB - Vascular endothelial growth factor A (VEGF-A) stimulates angiogenesis, but is also pro-inflammatory and plays an important role in the development of neurological disease, where it can have both attenuating and exacerbating effects. VEGF-B, a related molecule, is highly expressed in the central nervous system and seems to be important in neurological injury. A few studies have indicated that VEGF-A may play a role in the pathogenesis of multiple sclerosis (MS), but the role of VEGF-B has not been studied. We have studied the expression of VEGF-A, -B and their receptors by mRNA in situ hybridization, immunohistochemistry and real-time PCR in spinal cord from LEW rats with experimental autoimmune encephalomyelitis (EAE) and in cerebrospinal fluid (CSF) and blood samples from MS patients. Whereas VEGF-A is downregulated in glia in EAE, the infiltrating inflammatory cells are positive for VEGF-A. Expression of VEGF-B and the VEGF receptors is unaltered. In addition, the levels of VEGF-A mRNA in mononuclear cells [corrected] in CSF are lower in MS patients compared with controls. These results demonstrate a complex regulation of VEGF-A during neuroinflammation and suggest that VEGF-B is not involved in the pathogenesis of MS. PMID- 17083618 TI - Strain differences and the role for HSP47 and HSP70 in adjuvant arthritis in rats. AB - Because of high sequence homology between microbial and endogenous heat shock proteins (HSP), immunological cross-reactivity to microbial HSP has been suggested as a possible cause of the development of autoimmune diseases, such as rheumatoid arthritis. The present study aimed to determine a potential role of HSP47, a molecular chaperone involved in the synthesis and assembly of collagen molecules, and microbial HSP71 (mHSP71) in adjuvant arthritis (AA) in two rat strains: Dark Agouti (DA), susceptible to AA induction and Albino Oxford (AO), which is resistant to AA induction. Immunization with complete Freund's adjuvant (CFA) induced an increased expression of HSP47 in joints of DA rats, which exhibited severe clinical signs of AA at the time of disease peak, while this protein was not detectable in joints of AO rats. In contrast, no strain differences in HSP72 (rat analogue of mHSP71) expressions in joints were observed. The increased levels of anti-HSP47 antibodies were detected in sera of DA rats during the AA peak, while the immunization with CFA increased levels of anti-mHSP71 antibodies in sera of AO rats. HSP47 and mHSP71 reduced proliferation of draining inguinal lymph node cells (LNC) in resistant AO rat strain, leading to a hypothesis that both HSP participated in AA control. Finally, mHSP71 potentiated the apoptotic response of LNC in susceptible DA rat strain. In conclusion, our findings indicate involvement of HSP47 in the development of AA in the rat, and point out to the regulatory role for both HSP47 and mHSP71. PMID- 17083619 TI - Surfactant protein D of the innate immune defence is inversely associated with human obesity and SP-D deficiency infers increased body weight in mice. AB - Surfactant protein D (SP-D) is a key regulator of pathogen-induced inflammation. SP-D is further involved in lipid homeostasis in mouse lung and circulation and recent data have demonstrated that the body mass index (BMI; in kg/m(2)) is influenced by genes in common with SP-D. The objective of the present study was to describe the association between serum SP-D and weight, waist circumference or BMI, and furthermore to observe body weight development in SP-D-deficient (Spd-/ ) mice. As a part of the Danish population-based twin study (GEMINAKAR) on the metabolic syndrome, we analysed 1476 Danish twins for serum SP-D and investigated associations with weight, waist circumference and BMI by multiple regression analysis. Serum SP-D was significantly and inversely associated with weight (P = 0.001) and waist circumference in men (P < 0.001) and to BMI in both genders (P = 0.039 women, P < 0.001 men). The age-dependent increase in serum SP-D was most prominent in lean persons (BMI < 20). Spd-/- mice and wild-type mice were subjected to a feeding study and body weights were recorded in a time course over 24 weeks. Spd-/- mouse weight gain was significantly increased, with 90 mg/week (P < 0.0001) in males on normal chow. Fat percentage was significantly increased by 17% in the Spd-/- male mice (P = 0.003). We conclude, that there is an association between low levels or absent SP-D and obesity. PMID- 17083620 TI - Identification and characterization of secretagogin promoter activity. AB - Secretagogin is a newly identified calcium-binding protein selectively expressed in neuroendocrine tissue and pancreatic beta-cells. The function of secretagogin is unknown, but it has been suggested in beta-cells to influence calcium-influx, insulin secretion and proliferation, and has been observed downregulated in diabetes-prone BB rat islets exposed to cytokines. In the present study, we identified and characterized promoter activity of a human 1498 bp sequence upstream the transcription start site. The promoter sequence showed subtle but significant regulation by glucose within the normo-physiological range. Glucose also led to changes in expression of secretagogin protein in INS-1e cells, but not in primary cells from non-diabetes-prone Wistar Furth rats. No effects of cytokines neither on promoter activity nor protein expression were observed. The promoter region was furthermore screened by direct sequencing, and 11 polymorphisms were identified. Genotyping in a large homogenous Type 1 diabetes (T1D) family collection did not reveal association with T1D. PMID- 17083621 TI - Two neisseria meningitidis strains with different ability to stimulate toll-like receptor 4 through the MyD88-independent pathway. AB - Neisseria meningitidis causes acute severe diseases, including sepsis and meningitis, and more benign manifestations such as chronic meningococcemia or colonization of the upper respiratory tract. The inflammatory response, which contributes to the pathogenesis of meningococcal disease, is initiated by pattern recognition receptors, among which Toll-like receptors (TLR)s have been ascribed a particularly important role. We have previously demonstrated that N. meningitidis induce proinflammatory cytokine expression through TLR2 and TLR4. Here we characterize the molecular basis for differential activation of the inflammatory response by two N. meningitidis strains. This difference was due to differential ability to activate signal transduction through TLR4, as HEK293 cells expressing TLR4 produced significantly different levels of interleukin-8 in response to these strains. At the level of signal transduction, the two strains differed substantially in their ability to activate the pathway to nuclear factor kappaB in HEK293-TLR4/MD2 cells at late, but not early, time points. TLR4 activates two signal transduction pathways: one dependent on the adaptor molecule MyD88 and one independent of MyD88, and these pathways induce distinct patterns of gene expression in response to TLR4 ligands. By using macrophages from TLR2-/- mice, we observed that the two strains differed in their ability to activate the TLR4-induced MyD88-independent pathway, but not the MyD88-dependent pathway. This idea was further supported by experiments where either of the two pathways was inhibited and IL-8 secretion was measured. These data therefore provide molecular insight into activation of the inflammatory response by N. meningitidis, which is one of the key events in the pathogenesis of meningococcal disease. PMID- 17083622 TI - Levels of IL-15 in serum and cerebrospinal fluid of patients with Behcet's disease. AB - Interleukin-15 (IL-15) is a novel proinflammatory cytokine, involved in the pathogenesis of inflammatory/autoimmune disease. The objective of our study was to measure serum and cerebrospinal fluid (CSF) IL-15 levels in patients with Behcet's disease (BD). CSF/serum IL-15 ratio was introduced to assess the origin of elevated IL-15 levels. We measured serum and CSF-IL-15 levels in 40 patients with BD (20 patients in active stage). Inflammatory and non-inflammatory neurological disease patients acted as controls. Active BD patients have significantly higher serum IL-15 levels (median 10.4 pg/ml; range 5.3-17.4) compared with BD in remission (6.05 pg/ml; 4-10.4) and healthy controls (4.65 pg/ml; 3.9-6.2). Similar serum IL-15 levels were found in active neuro-BD and inflammatory neurological disease (9.5 pg/ml; 5-13). Elevated levels of IL-15 were observed in CSF samples from neuro-BD patients (11 pg/ml; 8.5-15) and inflammatory neurological disease patients (10 pg/ml; 6.5-14) compared with patients with non-inflammatory neurological disease (4 pg/ml; 4-5.5; P < 0.001). Vascular cerebral BD lesions were associated with high CSF/serum IL-15 ratio. Our findings suggest that IL-15 is involved in BD inflammatory process, particularly in vasculitis foci, as an elevated CSF/serum IL-15 ratio characterizes vascular cerebral lesions. PMID- 17083623 TI - Systemic antibody response to diarrheagenic Escherichia coli and LPS O111, O157 and O55 in healthy Brazilian adults. AB - Enterohaemorrhagic Escherichia coli (EHEC) can cause a variety of human illnesses ranging from uncomplicated diarrhoea to haemorrhagic colitis and haemolytic uremic syndrome. The serotype O157:H7 has been associated with numerous outbreaks worldwide, but in Brazil the infection is rare. Brazilian adults present antibodies reactive with the principal virulence factors of enteropathogenic E. coli (EPEC) that have many genetic and antigenic similarities with EHEC. Lipopolysaccharides (LPS) are components of outer membranes and important virulence factors of Gram-negative bacteria. LPS O111 is present in EPEC and EHEC strains. LPS O157 is found only in EHEC strains, but it has some structural similarities with LPS O55 present in EPEC strains. This study investigates the levels of IgG and IgM seric antibodies reactive with EHEC O157:H7, EHEC O111:H-, EPEC O111:H- and the levels of anti-LPS O111, LPS O157 and LPS O55 antibodies in healthy adults living in Sao Paulo, Brazil. The antibody levels were determined by an enzyme-linked immunosorbent assay for 100 individual serum samples, and the presence of anti-bacterial and anti-LPS seric antibodies was confirmed. Positive correlations were found among the three kinds of antibodies. The concentrations of IgM anti-LPS were significantly higher than those of IgG, and surprisingly, the concentrations of anti-LPS O157 were high in view of the infrequent isolation of O157 bacteria in Brazil. Our results suggest that there is a cross-reacting immunity to EHEC in the Brazilian population, which may be a result of the immunity to EPEC antigens. Alternatively, Brazilians may be exposed to EHEC more frequently than has previously been thought. PMID- 17083624 TI - Phenotypic analysis of IL-10-treated, monocyte-derived dendritic cells in patients with systemic lupus erythematosus. AB - Dendritic cells (DC) play a dual role in the immune response, participating in its induction, and the maintenance of immune tolerance. The aim of this work was to perform a quantitative and phenotypic analysis of DC generated in vitro in the presence of IL-10 in patients with systemic lupus erythematosus (SLE). Blood samples were obtained from 10 active and untreated patients with SLE and six controls. Monocyte-derived DC were generated in vitro in the presence or absence of IL-10, and a quantitative and phenotypic analysis was performed. We found that freshly isolated monocytes from SLE patients had an increased expression of CD11b. On the other hand, the efficiency of in vitro DC generation was diminished in blood samples from SLE patients for conventional DC, but not for IL-10-treated DC. A diminished expression of HLA-DR, CD9 and CD86 was observed in conventional DC from SLE patients compared with controls. In contrast, enhanced levels of HLA DR, CD80, CD9 and CD151 tetraspanins, FN1 (a class II MHC-tetraspanin epitope), CD85j/ILT2 and CD69 were detected in IL-10-treated DC from SLE patients. Accordingly, the phenotypic profile of IL-10-treated DC was very different in SLE and controls. However, the synthesis of IL-10 and IL-12 was similar in IL-10 treated and conventional cells in both SLE patients and controls. Our findings on the aberrant phenotype of IL-10-treated DC in SLE and their normal efficiency of in vitro generation may be important for the design of future therapies of this condition based on the administration of DC to induce immune tolerance. PMID- 17083625 TI - Binding characteristics of SLE anti-DNA autoantibodies to Catecholestrogen modified DNA. AB - Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease in which anti-double-stranded DNA antibody is a classic autoantibody that characterizes SLE. A role for oestrogens in the pathogenesis of SLE has been suspected for many years but the exact patho-aetiology remains elusive. In this study, the binding of SLE autoantibodies with native and 4-OHE(2)-NO-modified plasmid DNA were assessed. Binding specificity of antibodies was analysed by direct binding and inhibition enzyme-linked immunosorbent assay, quantitative precipitin titration and gel retardation assay. Anti-DNA IgG from SLE sera, purified on Protein A Agarose matrix, exhibited increased recognition of 4-OHE(2)-NO-DNA than native DNA (P < 0.001). Gel retardation assay further substantiated the enhanced recognition of modified DNA by anti-DNA autoantibodies. The affinity of anti-DNA antibodies for modified polymer was found to be high as calculated by using Langmuir plot. DNA modified by 4-OHE(2)-NO presents unique neo-epitopes that might be one of the factor in antigen-driven induction of SLE autoantibodies. PMID- 17083626 TI - Prevalence of IgG antibody against measles, mumps and rubella in bangladeshi children: a pilot study to evaluate the need for integrated vaccination strategy. AB - The present study was carried out to determine the seroprevalence of IgG antibodies in Bangladeshi children against measles (irrespective of vaccination status), mumps and rubella (MMR) to assess strategic need of combined vaccination for these diseases. A total of 456 children of 1 month to 15 years, were studied. Serum IgG antibodies against MMR were measured by enzyme-linked immunosorbent assay (ELISA). By 3 months, protective IgG antibody level (>40 AU for measles and mumps and >15 IU/ml for rubella) for the diseases found to be between 50% and 80% among the studied children. Protective measles antibody (IgG) was not detected in all the children of 3-9 months and significant number of children between 9 months and 5 years were unprotected (87-65%; P < 0.001). Moreover, children of 3 15 months had no protective antibody level against mumps and significant number of children between 15 months and 5 years were unprotected (92-71%; P < 0.001). Between 5 and 15 years of age, significant number of children became protective (63-85%, P < 0.001). Although, a majority of children between 3 months and 5 years had shown to have no protective antibody against rubella (89-71%; P < 0.01 0.001) between >10 and 15 years 71% children had protective level of antibodies (P < 0.001). No significant difference was observed in antibody prevalence regarding socioeconomic classes, nutritional status and parental education. The data showed that: (i) a significant number of children remain unprotected against MMR in childhood and (ii) an extensive nationwide survey is required to suggest an integrated vaccination strategy in order to implement appropriate control measures of the three infectious diseases. PMID- 17083627 TI - Acute suppression of serum IgM and IgA in tank workers exposed to benzene. AB - We investigated associations between benzene exposure and alterations of proteins and cells of the immune system among workers maintaining cargo tanks containing crude oil residues. Individual exposure to benzene, benzene in blood and urine, peripheral blood lymphocytes (total lymphocytes, lymphocytes in subpopulations CD3, CD4, CD8, CD19, CD56 and CD4/CD8 ratio), complement factors C3 and C4 and serum concentration of immunoglobulins (IgG, IgA, IgM and IgE) were analysed among 13 tank workers and nine unexposed referents (catering section). Benzene exposure was measured during three consecutive 12-h work days. Blood and urine samples were collected pre-shift on the first day (baseline), post-shift on the third day, and pre-next shift on the following morning. The time spent in the cargo tank was logged. The individual geometric mean benzene exposure in the breathing zone of tank workers over 3 days was 0.15 p.p.m. (range 0.01-0.62 p.p.m.) (n = 26). The geometric mean benzene concentration in blood post-shift was 12.3 nmol/l among tank workers versus 0.7 nmol/l among the referents. Tank workers showed a decline (versus referents) in IgM from baseline to post-shift (t test, P = 0.04) and IgA from baseline to pre-next shift (t-test, P = 0.01). They also showed a decline in CD4 T cells from baseline to post-shift (t-test, P = 0.04). Suppression correlated with benzene exposure, benzene concentrations in blood and urine and time spent in the tank. The groups did not differ significantly in the change in other immune parameters. The clinical significance is unknown and warrants further studies. PMID- 17083630 TI - Avian influenza: a pandemic waiting in the wings? AB - Recent widespread outbreaks of avian influenza and, associated with these a growing number of human infections with a high mortality rate, have raised concerns that this might be the prelude to a severe pandemic of human influenza. As a background to these concerns the present article reviews influenza as a human disease, its origins and the involvement of other species, properties of the influenza viruses and the current status of influenza prevention and control. PMID- 17083631 TI - Avian influenza pandemic threat and health systems response. AB - Avian influenza is a panzootic and recurring human epidemic with pandemic potential. Pandemic requirements for a viral pathogen are: a novel virus must emerge against which the general population has little or no immunity; the new virus must be able to replicate in humans and cause serious illness; and the new virus must be efficiently transmitted from person to person. At present, only the first two conditions have been met. Nonetheless, influenza pandemics are considered inevitable. Expected worldwide human mortality from a moderate pandemic scenario is 45 million people or more than 75% of the current annual global death burden. Although mathematical models have predicted that an emerging pandemic could be contained at its source, this conclusion remains controversial among public health experts. The Terrestrial Animal Health Code and International Health Regulations are enforceable legal instruments integral to pandemic preparedness. Donor support in financial, material and technical assistance remains critical to disease control efforts - particularly in developing countries where avian influenza predominately occurs at present. Personal protective equipment kits, decontamination kits and specimen collection kits in lightweight, portable packages are becoming standardized. Air transport border control measures purporting to delay importation and spread of human avian influenza are scientifically controversial. National pandemic plans prioritize beneficiary access to antiviral drugs and vaccines for some countries. Other medical commodities including ventilators, hospital beds and intensive care units remain less well prioritized in national plans. These resources will play virtually no role in care of the overwhelming majority of patients worldwide in a pandemic. Prehospital care, triage and acute care all require additional professional standardization for the high patient volumes anticipated in a pandemic. PMID- 17083632 TI - Multislice computed tomographic pulmonary angiography for diagnosing pulmonary embolism in the emergency department: has the 'one-stop shop' arrived? AB - Current diagnostic pathways for pulmonary embolism are complex and involve multiple tests. Clinicians await a single diagnostic modality that accurately rules in or out pulmonary embolism and also provides additional diagnostic and prognostic information. Multislice computed tomographic pulmonary angiography (msCTPA) might be that test. msCTPA has good outcomes using current reference standards and can be used as a stand alone test in low risk patients. Additional leg vein imaging should still be performed for higher-risk patients. CT venography performed immediately after msCTPA has sufficient sensitivity and specificity for proximal deep venous thrombosis to allow a single scan to rule out venous thromboembolism in most patients. Up to 40% of patients have important alternative diagnoses seen on msCTPA. Right ventricular measurements may be calculated from msCTPA data and provide prognostic information for patients with severe PE. This may replace the need for echocardiography in the future. msCTPA provides a safe single radiological investigation for diagnosing PE for most patients, after risk stratification and D-dimer testing. A pathway is provided for msCTPA use in the management of PE in the ED. PMID- 17083633 TI - Training of disaster managers at a masters degree level: from emergency care to managerial control. AB - The world has faced huge disasters over the last few decades and concerns have been expressed by nearly all international agencies involved that there is a scarcity of managerial skills to deal with the mitigation and management of disasters. Disaster risks are also on the increase throughout Africa and Southern Africa because of changes in the development process, settlement patterns and conflicts in the region. Emergency physicians are but one important resource in dealing with disasters. The need for a comprehensive multisectoral approach to disasters and more importantly to deal with its mitigation is becoming increasingly evident, especially in developing countries. Hence, the need for specially trained professionals in disaster management. In an effort to improve national, regional and continental capacity, and in support of the South African Disaster Management Act, the University of the Witwatersrand, Johannesburg, South Africa, has developed a Master of Public Health degree in Disaster Management. The MPH is aimed at preparing professionals from health and allied fields to play leadership roles in the management, improvement and evaluation of health and the health-care system. Emergency physicians have an important role to play in the development of disaster medicine and disaster management programmes and it is important that they engage in this activity, collaborating with colleagues of various other disciplines as appropriate. The following paper outlines the background to the programme and the current programme. PMID- 17083634 TI - Does an immunochromatographic D-dimer exclude acute lower limb deep venous thrombosis? AB - BACKGROUND: A pre-test probability score and D-dimer may reduce the need for ultrasound examinations for excluding lower limb deep venous thrombosis (DVT). OBJECTIVE: To establish the accuracy of an immunochromatographic D-dimer assay called 'Simplify' for diagnosis of acute DVT by complete (calf veins included) lower limb ultrasound examination. METHODS: A total of 453 consecutive patients presented to the ED of a tertiary centre with suspected first episode of DVT, were prospectively recruited. A pre-test probability score (Hamilton Score), an immunochromatographic D-dimer and a complete, single, unilateral lower limb ultrasound examination were performed in all patients. All patients with a negative ultrasound examination were followed up for 3 months. RESULTS: There were 159 men and 294 women with a mean age of 55.8 years (SD 20.3). Of the 227 patients with a negative D-dimer, 214 patients had negative ultrasound examinations and 13 patients had isolated calf DVT. Among the 226 patients with a positive D-dimer, 74 patients had DVT and 152 patients had negative ultrasound examinations. The sensitivity, specificity, positive and negative predictive values were 85.1% (75.8-91.8), 58.5% (53.4-63.5), 32.7% (26.6-38.9) and 94.3% (90.4-96.9), respectively. One hundred and sixty-five patients had an unlikely Hamilton Score and a negative D-dimer. The negative predictive value of the immunochromatographic D-dimer in an unlikely Hamilton Score population was 98.8% (95.7-99.8%). CONCLUSION: An unlikely probability Hamilton Score and a negative immunochromatographic D-dimer reliably exclude both proximal and isolated calf DVT. PMID- 17083635 TI - Emergency Department Communication Officer: response to consumer demand. AB - OBJECTIVE: To determine whether the employment of a Communication Officer (CO) in the ED waiting room improves ED consumer satisfaction with regard to information exchange and general care received in the waiting room. METHODS: Multifactorial analytic cross sectional survey on convenience samples of ED consumers. Consumers were eligible if they had been in the waiting room for more than 30 min. Comparisons in responses using ordinal regression analysis were made between shifts on which there was no intervention present, where the CO, an educational video or both were present. RESULTS: When the CO was present the responses to seven of 10 questions regarding information exchange and general care were significantly improved (all P < 0.01) and three were not. When only the video was running, the responses to none of the 10 questions were significantly improved. The concurrent presence of the video neither enhanced nor detracted from the responses regarding the CO when both were present. Responses to all five questions regarding the CO were positive, and responses to two of three questions regarding the value of the video were positive. CONCLUSION: The presence of a designated CO in the ED waiting room enhances the communication with, and perceptions of general care by ED consumers. PMID- 17083636 TI - State of play: clearing the thoracolumbar spine in blunt trauma victims. AB - INTRODUCTION: The present article reviews the clinical and imaging clearance of the thoracic and lumbar spines of blunt trauma victims and the evolution of these strategies with the use of new imaging technologies. METHODS: A comprehensive literature search was performed, and articles identified were critically appraised RESULTS: Twenty papers were identified, 12 directly comparing computed tomography screening with plain X-rays. Evidence-based protocols are presented. The evolution of imaging strategies in response to new technology is described. CONCLUSIONS: Thoracolumbar spine screening is best done using reformatted images acquired when scanning the chest and abdomen of high-risk multi-trauma patients. If computed tomography is not clinically indicated for investigation of other injuries then plain films are the first line investigation. PMID- 17083637 TI - Randomized comparison of the SLIPA (Streamlined Liner of the Pharynx Airway) and the SS-LM (Soft Seal Laryngeal Mask) by medical students. AB - OBJECTIVE: The aim of the study was to compare the Streamlined Liner of the Pharynx Airway (SLIPA; Hudson RCI), a new supraglottic airway device, with the Soft Seal Laryngeal Mask (SS-LM; Portex) when used by novices. METHODS: Thirty six medical students with no previous airway experience, received manikin training in the use of the SLIPA and the SS-LM. Once proficient, the students inserted each device in randomized sequence, in two separate patients in the operating theatre. Only two insertion attempts per patient were allowed. Students were assessed in terms of: device preference; success or failure; success at first attempt and time to ventilation. RESULTS: Sixty-seven per cent of the students preferred to use the SLIPA (95% confidence interval 49-81%). The SLIPA was successfully inserted (one or two attempts) in 94% of patients (34/36) and the SS-LM in 89% (32/36) (P = 0.39). First attempt success rates were 83% (30/36) and 67% (24/36) in the SLIPA and SS-LM, respectively (P = 0.10). Median time to ventilation was shorter with the SLIPA (40.6 s) than with the SS-LM (66.9 s) when it was the first device used (P = 0.004), but times were similar when inserting the second device (43.8 s vs 42.9 s) (P = 0.75). CONCLUSIONS: In the present study novice users demonstrated high success rates with both devices. The SLIPA group achieved shorter times to ventilation when it was the first device they inserted, which might prove to be of clinical significance, particularly in resuscitation attempts. Although the Laryngeal Mask has gained wide recognition for use by both novice users and as a rescue airway in failed intubation, the data presented here suggest that the SLIPA might also prove useful in these areas. PMID- 17083638 TI - Randomized, single blinded control trial comparing haemolysis rate between two cannula aspiration techniques. AB - OBJECTIVE: Blood aspiration through i.v. cannulae is a common ED sampling method. It is, however, associated with increased haemolysis rates when compared with direct venipuncture. Our ED utilizes two common methods of sampling through cannulae. The aim of the present study was to examine the haemolysis rates for these two techniques. METHODS: A randomized, single blinded prospective study was performed. From a convenience sample patients were randomized to either 'Method one' (blood taken directly from cannula hub) or 'Method two' (blood taken using an Interlink device through cannula cap). For each group patient demographics, clinical information and haemolysis were analysed. chi(2)-analysis and a multivariate logistic regression model were applied to determine significant relationships. RESULTS: A total of 1390 patients were enrolled, 694 had blood collected by 'Method one' and 696 by 'Method two', with haemolysis rates of 6.5% and 7.2%, respectively. Univariate analysis showed ease of aspiration to be the only variable to have a strong association with haemolysis (P < 0.0001). After allowing for ease of aspiration the technique used did not significantly affect haemolysis rate (P = 0.7). CONCLUSIONS: The rate of haemolysis is unrelated to the technique of blood aspiration. However, haemolysis rates increase significantly with perceived difficulty of blood draw. PMID- 17083639 TI - Development and implementation of an education and credentialing programme to provide safe paediatric procedural sedation in emergency departments. AB - OBJECTIVE: In the conduct of paediatric procedural sedation (PPS) within the ED the combination of powerful drugs, variable competency levels and high staff turnover carry the potential for sedation-associated adverse events. Yet, currently, there is no set programme for education and accreditation of ED staff in PPS. We set out to develop such a programme. METHODS: We outline the development process of a comprehensive multidisciplinary PPS programme and present its key educational elements (sedation manual, lecture, treatment order form and checklist, parent information handout) and credentialing through multiple-choice questions and competency assessments. We describe issues associated with the implementation of the programme at a metropolitan mixed ED and the ED of a major tertiary paediatric centre. RESULTS: Since its inception a total of 294 emergency staff have either completed or have partially completed the programme. Staff feedback showed that the majority of staff scored the elements of the programme as very good to excellent, and felt that their sedation skills had improved and their practice was safer. The development and implementation of the PPS programme raised many issues and posed a number of challenges. We describe the strategies we used to overcome such challenges and barriers. CONCLUSION: We present the development and implementation of a comprehensive PPS programme for emergency staff. As a result of the multicentre development process involving a community and a tertiary paediatric ED the programme will likely have broad applicability in different types of ED caring for children. PMID- 17083640 TI - Evaluation of the impact of a paediatric procedural sedation credentialing programme on quality of care. AB - OBJECTIVES: The aim of the present study is to describe changes in documentation, risk assessment and patient care resulting from implementation of a credentialing process for medical and nursing staff in paediatric procedural sedation (PPS) in two EDs - one an urban mixed ED and the other a specialist paediatric ED. METHODS: Chart review of 100 patients undergoing PPS prior to and 100 patients following introduction of the PPS programme. Information was extracted from medical records and sedation checklists. Demographics, drugs used, procedure performed and elements of the pre-procedural, intra-procedural and post procedural care were compared pre- and post implementation of the PPS programme. RESULTS: Significant improvements in the post-implementation period compared with pre-implementation were seen in: frequency of documentation of informed consent (87 vs 15%, P < 0.0001); evidence of performance of a pre-procedural risk assessment (87 vs 1%, P < 0.0001); and appropriate recording of vital signs (58 vs 27%, P < 0.0001). Improvements were also noted in documentation of weight, allergies, fasting status and recording of drug orders. There were no adverse events recorded in the pre-programme period and 6 recorded in the post-programme period. CONCLUSION: The implementation of a PPS credentialing programme into these two EDs resulted in significant improvements in risk assessment, monitoring and documentation of important information related to safe PPS. These improvements should result in improved quality and safety of PPS. PMID- 17083641 TI - Levosimendan: a new option in acute cardiac failure. PMID- 17083642 TI - On burnout and other demons. AB - A personal perspective on the subject of burnout in emergency physicians. Causes, identification of and potential pitfalls as well as possible remedies are discussed. PMID- 17083643 TI - So close and yet so far: health care in our nearest neighbour. AB - Emergency medicine is reaching out from traditional westernized birth places to the uncharted territories of the developing world. With ever-increasing links with our near neighbours in Asia, a steady stream of ACEM fellows and trainees are giving their time and resources in aid of their health sector development. Papua New Guinea, being our nearest neighbour, deserves our attention not only for the stark contrast to our own health system experience, but also because of the rising specialty of emergency medicine in that country. I was lucky enough to be part of a concerted programme to train medical students and emergency medicine trainees in the fundamentals of our specialty. PMID- 17083645 TI - Fall about laughing: a case of laughter syncope. AB - Laughter syncope is an unusual but recognized form of situational syncope likely to have a similar pathophysiological origin to tussive syncope. There are few case reports of this syndrome in the literature. Patients, as in this case, might present initially to the ED, and laughter should be considered among the numerous differentials for syncope. PMID- 17083646 TI - Stroke thrombolysis: still no consensus. PMID- 17083647 TI - Trp207Gly in platelet glycoprotein Ibalpha is a novel mutation that disrupts the connection between the leucine-rich repeat domain and the disulfide loop structure and causes Bernard-Soulier syndrome. AB - BACKGROUND: Bernard-Soulier syndrome (BSS) is a severe inherited bleeding disorder that is caused by a defect in glycoprotein (GP)Ib-IX-V complex, the platelet membrane receptor for von Willebrand factor. PATIENTS: The diagnosis of BSS was made in two members of a Bukharian Jewish family who had life-long thrombocytopenia associated with mucocutaneous bleeding manifestations. METHODS AND RESULTS: Flow cytometry and Western blot analyses showed only trace amounts of GPIb and GPIX on the patients' platelets. Sequence analysis of the GPIbalpha gene revealed a homozygous T > G transversion at nucleotide 709 predicting Trp207Gly substitution in the mature protein. Introduction of the mutation into a mammalian expression construct abolished the surface expression of GPIbalpha in transfected baby hamster kidney cells. The crystal structure of the N-terminus of GPIbalpha (PDB: 1SQ0) indicates that Trp207 is completely buried and located in a disulfide loop structure that interacts with the leucine-rich repeat (LRR) domain. CONCLUSION: A novel mutation, Trp207Gly, causes BSS and predicts disruption of the interaction between a disulfide loop and the LRR domain that is essential for the integrity of GPIbalpha structure. PMID- 17083648 TI - Effects of exposure to air pollution on blood coagulation. AB - BACKGROUND: Consistent evidence has indicated that air pollution increases the risk of cardiovascular diseases. The underlying mechanisms linking air pollutants to increased cardiovascular risk are unclear. OBJECTIVES: We investigated the association between the pollution levels and changes in such global coagulation tests as the prothrombin time (PT) and the activated partial thromboplastin time (APTT) in 1218 normal subjects from the Lombardia Region, Italy. Plasma fibrinogen and naturally occurring anticoagulant proteins were also evaluated. METHODS: Hourly concentrations of particulate (PM10) and gaseous pollutants (CO, NO2, SO2, and O3) were obtained from 53 monitoring sites covering the study area. Generalized additive models were applied to compute standardized regression coefficients controlled for age, gender, body mass index, smoking, alcohol, hormone use, temperature, day of the year, and long-term trends. RESULTS: The PT became shorter with higher ambient air concentrations at the time of the study of PM10 (coefficient = -0.06; P < 0.05), CO (coefficient = -0.11; P < 0.001) and NO2 (coefficient =-0.06; P < 0.05). In the 30 days before blood sampling, the PT was also negatively associated with the average PM(10) (coefficient = -0.08; P < 0.05) and NO2 (coefficient = -0.08; P < 0.05). No association was found between the APTT and air pollutant levels. In addition, no consistent relations with air pollution were found for fibrinogen, antithrombin, protein C and protein S. CONCLUSIONS: This investigation shows that air pollution is associated with changes in the global coagulation function, suggesting a tendency towards hypercoagulability after short-term exposure to air pollution. Whether these changes contribute to trigger cardiovascular events remains to be established. PMID- 17083649 TI - IgG classification of anti-PF4/heparin antibodies to identify patients with heparin-induced thrombocytopenia during mechanical circulatory support. AB - Commercial immunoassays frequently detect anti-PF4/heparin antibodies during mechanical circulatory support (MCS), but only a small minority of patients develops heparin-induced thrombocytopenia (HIT). Whereas platelet functional tests can distinguish between platelet-activating and non-platelet-activating antibodies, commercial PF4-dependent immunoassays do not. Between 2003 and 2004, 113 patients were placed on MCS. Blood samples were obtained on postimplant day 5 7 for analyses by antibody assays and the functional heparin-induced platelet activation (HIPA) assay. Three distinct groups of patient sera were identified: platelet-activating anti-PF4/heparin antibodies (n = 10), non-platelet-activating anti-PF4/heparin antibodies (n = 53), and anti-PF4/heparin antibody negative (n = 50). Patients with platelet-activating antibodies had the highest risk for thromboembolic events (P < 0.005), whereas those with non-platelet-activating antibodies did not differ from antibody negative patients (P = 0.369). The enzyme immunoassay and column agglutination assays, which cover all immunoglobulin classes, demonstrated adequate sensitivity and negative predictive value; yet, both lacked specificity with respect to the platelet-activating antibodies. If all antibody positive patients were further classified by an IgG-specific anti PF4/heparin enzyme-immuno assay, specificity for platelet-activating antibodies increased. Whereas IgG-specific optical density (OD) values below 1.0 were likely for non-platelet-activating anti-PF4/heparin antibodies, higher values were progressively predictive for pathogenic platelet activation. The probability of the development of clinical HIT also increased steeply. In conclusion, platelet activating anti-PF4/heparin antibodies are relatively common (about 9%) in patients on MCS and are associated with significantly higher thrombotic event rates. Low IgG-specific OD values (< 1.0) in the enzyme-immunoassay indicate low likelihood for the presence of platelet-activating antibodies. These results justify further validation so that anticoagulation during MCS becomes safer and adequate. PMID- 17083650 TI - A new multiplex PCR and conformation-sensitive gel electrophoresis strategy for mutation detection in the platelet glycoprotein alphaIIb and beta3 genes. PMID- 17083651 TI - Maternal urinary tract infection and related drug treatments during pregnancy and risk of congenital abnormalities in the offspring. AB - OBJECTIVE: The association between urinary tract infection (UTI) of pregnant women and preterm birth/low birthweight is known, but the possible association between UTI and congenital abnormalities (CAs) was evaluated rarely. Only one study showed an association with atrial septal defect, thus we decided to check this possible association. DESIGN: The population-based large data set of the Hungarian Case-Control Surveillance of Congenital Abnormalities (HCCSCA); most maternal UTIs were based on medically recorded data. SETTING The HCCSCA, 1980 1996, contained 22 843 newborns or fetuses with CAs and 38 151 matched controls, i.e. newborn infants without any HCAs. POPULATION: Hungarian informative offspring: live births, stillbirths and prenatally diagnosed malformed fetuses. METHODS: Case-control pair analysis. MAIN OUTCOME MEASURES: Twenty-five CA groups. RESULTS: A total of 1542 (6.75%) mothers in the case group had UTI during entire pregnancy compared with 2188 (5.74%) mothers in the control group (adjusted prevalence odds ratios [POR] with 95% CI: 1.15, 1.06-1.24). We did not find a higher prevalence of UTI during the second and/or third months of pregnancy in total case group (adjusted POR with 95% CI: 1.1, 0.9-1.2) and in any group of CAs including atrial septal defect type II. CONCLUSIONS: No evidence for the teratogenic effect of maternal UTI and related drug treatments during early pregnancy. PMID- 17083652 TI - Sublingual compared with vaginal misoprostol for labour induction at term: a randomised controlled trial. AB - OBJECTIVE: To compare the efficacy and safety of 50 microg of sublingual misoprostol with 25 microg of vaginal misoprostol administered for labour induction at term. Design Double-blinded, randomised controlled trial. Setting University Hospital, Kaunas, Lithuania. Sample A total of 140 women at term with indications for labour induction. Methods Women were randomised to receive either 50 microg of sublingual misoprostol with vaginal placebo (n = 70) or sublingual placebo with 25 microg of vaginal misoprostol (n = 70) every 4 hours (maximum six doses). Main outcome measures The number of women delivering vaginally within 24 hours of labour induction. Results Fifty-eight women (83%) in the sublingual misoprostol group and 53 (76%) in the vaginal misoprostol group delivered vaginally within 24 hours [relative risk (RR) 1.1, 95% confidential interval (CI) 0.9-1.3]. However, the induction to vaginal delivery time was significantly shorter in the sublingual group (15.0 +/- 3.7 hours) compared with the vaginal group (16.7 +/- 4.1 hours, P = 0.03). The incidence of tachysystole was more than three-fold higher in the sublingual than in the vaginal group (14 versus 4.3%; RR 3.3, 95% CI 0.9-11.6), but this was not statistically significant. There were no significant differences in the incidence of hypertonus or hyperstimulation syndrome, mode of delivery, interventions for fetal distress or neonatal outcomes between the two groups. Conclusion A 50 microg of sublingual misoprostol 4 hourly for labour induction at term seems to have similar efficacy as 25 microg of vaginal misoprostol. Further studies on safety with larger numbers of women need to be conducted before routine sublingual misoprostol use in this setting. PMID- 17083653 TI - Precancerous changes in the cervix and risk of subsequent preterm birth. AB - OBJECTIVE: The aims of this study were (i) to examine whether women referred for assessment of precancerous changes in the cervix had higher rates of preterm birth compared with those in the general population and (ii) to compare preterm birth rates for treated and untreated women adjusting for possible confounding factors. DESIGN: Retrospective cohort design. SETTING: Teaching hospital. POPULATION: All women referred to the Royal Women's Hospital, Melbourne (1982 2000), who subsequently had a birth recorded on the Victorian Perinatal Data Collection system (n = 5548). METHODS: Record linkage of hospital dysplasia clinic records and population-based birth records. MAIN OUTCOME MEASURES: Total preterm delivery (<37 weeks of gestation) and subtypes. RESULTS: Both treated and untreated women were at a significantly increased risk for preterm birth compared with those in the general population: treated--standardised prevalence ratio (SPR) 2.0, 95% CI 1.8-2.3 and untreated--SPR 1.5, 95% CI 1.4-1.7. Within the cohort, the treated women were significantly more likely to give birth preterm (adjusted OR 1.23, 95% CI 1.01-1.51). An increased risk of preterm birth was also associated with a history of induced or spontaneous abortions, illicit drug use during pregnancy or a major maternal medical condition. Cone biopsy, loop electrosurgical excision procedure and diathermy were associated with preterm birth. After adjusting for possible confounding factors, only diathermy remained significant (adjusted OR 1.72, 95% CI 1.36-2.17). Women treated using laser ablation were not at an increased risk for preterm birth (adjusted OR 1.1, 95% CI 0.8-1.4). CONCLUSIONS: Diagnosis of precancerous changes in the cervix (regardless of the treatment) was associated with an increased risk of preterm birth. Consideration should be given to the preferential use of ablative treatments. PMID- 17083654 TI - Lower urinary tract injuries after transobturator tape insertion by different routes: a large retrospective study. AB - OBJECTIVES: To identify the rate of, and risk factors for, lower urinary tract (LUT) injuries associated with the transobturator tension-free vaginal tape (TOT) procedure. DESIGN: Retrospective cohort study. SETTING: Tertiary referral urogynaecology centre. POPULATION: 390 women who underwent transobturator suburethral tapes for management of urodynamic stress incontinence between July 2002 and January 2006. METHODS: Early cases were identified from theatre records and a case note review performed. From May 2005 (n = 94), data from an ongoing prospective audit were reviewed. Data for LUT injuries with TOT procedures were examined and routes of insertion were compared using Fischer's exact test. MAIN OUTCOME MEASURES: Rate of LUT injuries associated with the TOT procedures. Assessment of factors increasing risk of LUT injury, and comparison of the 'outside-in' and 'inside-out' techniques. RESULTS: 241 women underwent TOT outside-in technique and 148 of them underwent inside-out technique. Four LUT injuries occurred (1%): two urethral injuries (0.5%) and two bladder injuries (0.5%). All LUT injuries occurred in the outside-in group, although this difference did not reach significance (P = 0.146). Bladder injuries occurred in women who underwent concomitant vaginal surgery, while urethral injuries occurred in women undergoing secondary procedures. CONCLUSION: LUT injury is an uncommon complication of the TOT procedures, and in our hands only occurred with the outside-in technique. Intraoperative cystoscopy should be considered only in selected cases. PMID- 17083655 TI - Impact and risk factors for early-onset group B streptococcal morbidity: analysis of a national, population-based cohort in Sweden 1997-2001. AB - OBJECTIVES: To study early-onset group B streptococcal (EOGBS) morbidity, mortality, and maternal risk factors. DESIGN: Observational, population-based, retrospective. Data from national registers and medical records. SETTING: Sweden, 1997-2001. POPULATION: Cohort of 640 infants with a diagnosis of GBS infection out of 435 070 live births. METHOD: Infants with diagnoses GBS sepsis (P36.0) and/or pneumonia (P23.3) were analysed. In cases with P36.0, register data were validated against infant and maternal medical records. Odds ratios (OR) were estimated by multiple logistic regression. MAIN OUTCOME MEASURES: Incidence of EOGBS morbidity, mortality, frequency of maternal risk factors and administration of intrapartum antibiotics. RESULTS: There were 319 cases with EOGBS sepsis. Blood culture verified 174 cases. There were 145 with clinical sepsis and 180 with pneumonia only. The incidences were 0.40, 0.33, and 0.41 per 1000 live births, respectively. The mortality was 7.5, 0.7, and 2.2% in respective groups. The frequencies of established maternal risk factors were: membrane rupture > or =18 hours, 44%; prematurity, 26%; temperature during labour > or =38 degrees C, 22%. Novel maternal risk factors identified in verified cases were gestational age (GA) of 37 completed weeks (OR 3.5, 1.8-6.5) and gestational diabetes (OR 3.7, 1.8-8.5). When including clinical sepsis, also epidural anaesthesia, infant large for GA, postmaturity, and high maternal age were significant risk factors. CONCLUSION: The incidence of verified EOGBS disease was 0.4 per 1000 live births with a total burden of EOGBS morbidity approximately three times higher. GA of 37 completed weeks and gestational diabetes were identified as additional significant risk factors. PMID- 17083656 TI - Personality and self-regulation: trait and information-processing perspectives. AB - This article introduces the special issue of Journal of Personality on personality and self-regulation. The goal of the issue is to illustrate and inspire research that integrates personality and process-oriented accounts of self-regulation. The article begins by discussing the trait perspective on self regulation--distinguishing between temperament and personality accounts--and the information-processing perspective. Three approaches to integrating these perspectives are then presented. These range from methodological approaches, in which constructs representing the two perspectives are examined in integrated statistical models, to conceptual approaches, in which the two perspectives are unified in a holistic theoretical model of self-regulation. The article concludes with an overview of the special issue contributions, which are organized in four sections: broad, integrative models of personality and self-regulation; models that examine the developmental origins of self-regulation and self-regulatory styles; focused programs of research that concern specific aspects or applications of self-regulation; and strategies for increasing the efficiency and effectiveness of self-regulation. PMID- 17083657 TI - Personality reflected in a coherent idiosyncratic interplay of intra- and interpersonal self-regulatory processes. AB - This article discusses a framework that conceptualizes personality in terms of a unique pattern of interacting intra- and interpersonal self-regulatory mechanisms employed in the service of constructing and maintaining a desired self. These personal goals motivate the individuals' self-construction efforts and give direction, organization, and coherence to the self-regulatory dynamics--both within the person and in the social world in which they play out. The framework is illustrated through research on construct validation of the narcissistic personality type and extended by brief applications to dependency and rejection sensitivity to show how it may help us understand the complex signatures that are the expressions of a personality type. It offers a guide for where to look for and how to organize the unique features and idiosyncratic dynamics of different self-construction types and to make sense of their otherwise often seemingly paradoxical expressions. In so doing, the framework speaks to basic goals of personality psychology by providing an approach for capturing trait-like individual differences while simultaneously shedding light on the psychological mechanism that underlies them. PMID- 17083658 TI - Self-regulation and the problem of human autonomy: does psychology need choice, self-determination, and will? AB - The term autonomy literally refers to regulation by the self. Its opposite, heteronomy, refers to controlled regulation, or regulation that occurs without self-endorsement. At a time when philosophers and economists are increasingly detailing the nature of autonomy and recognizing its social and practical significance, many psychologists are questioning the reality and import of autonomy and closely related phenomena such as will, choice, and freedom. Using the framework of self-determination theory (Ryan & Deci, 2000), we review research concerning the benefits of autonomous versus controlled regulation for goal performance, persistence, affective experience, quality of relationships, and well-being across domains and cultures. We also address some of the controversies and terminological issues surrounding the construct of autonomy, including critiques of autonomy by biological reductionists, cultural relativists, and behaviorists. We conclude that there is a universal and cross developmental value to autonomous regulation when the construct is understood in an exacting way. PMID- 17083659 TI - Children's conscience and self-regulation. AB - We focus on children's conscience, an inner guiding system responsible for the gradual emergence and maintenance of self-regulation. Drawing from our research program that has encompassed three large longitudinal studies cumulatively covering the first 6 years of life, we discuss two major components of conscience: moral emotions (guilt, discomfort following transgressions) and moral conduct compatible with rules and standards. We discuss the organization of young children's conscience, focusing on relations between moral emotions and moral conduct, and the development of conscience, focusing on its early form: the child's eager, willing stance toward parental socialization. We also review research on two major sets of influences that predict individual differences in moral emotions and moral conduct: biologically based temperament and socialization in the family. We discuss two inhibitory systems of temperament fearfulness and effortful control-and several features of socialization, including the style of parental discipline and the quality of the parent-child relationship. Early conscience is an important early personality system, coherently organized, relatively stable over time, and subject to individual differences that emerge as a result of a complex interplay between children's temperamental individuality and socialization in the family. PMID- 17083660 TI - The development of children's ideal and ought self-guides: parenting, temperament, and individual differences in guide strength. AB - Regulatory focus theory (RFT; Higgins, 1997) predicts that individual differences in the strength of promotion (ideal) and prevention (ought) orientations emerge from patterns of parent/child interactions that emphasize making good things happen versus keeping bad things from happening. This article examines the development of individual differences in the strength of children's promotion and prevention goals and presents selected findings from three studies exploring the origins of regulatory focus. We found a three-factor structure for parenting behaviors that differentiated between the presence/absence of positive outcomes versus the presence/absence of negative outcomes in two different data sets and validated that factor structure by examining its associations with maternal temperament. In turn, the parenting factors predicted individual differences in children's orientations to ideal and ought guides, and those associations were moderated by individual differences in child temperament. PMID- 17083661 TI - Strength and safety in numbers: considering the social implications of regulatory focus. AB - The present article examines how individual and situational differences in individuals' regulatory focus on nurturance and gain (promotion) and on security and safety (prevention) may have significant, and distinct, social and interpersonal implications. We first review recent research examining how significant others affect goal pursuit and how individual differences in regulatory focus may moderate the various behavioral, evaluative, and experiential manifestations of social identification. We then consider how regulatory focus moderates the way in which people "size up" their social world in terms of the efficiency in which they identify and appraise motivationally relevant aspects of their social environment. Finally, we explore how regulatory focus moderates people's deliberate and automatic reactions to the beliefs, expectations, behavior, and emotions of other individuals and social groups. PMID- 17083662 TI - Future orientation in the self-system: possible selves, self-regulation, and behavior. AB - Possible selves are representations of the self in the future. Early theoretical accounts of the construct suggested that possible selves directly influence motivation and behavior. We propose an alternative view of possible selves as a component in self-regulatory processes through which motivation and behavior are influenced. We demonstrate the advantages of this conceptualization in two studies that test predictions generated from theoretical models of self regulation in which the possible selves construct could be embedded. In one study, we show how viewing possible selves as a source of behavioral standards in a control-process model of self-regulation yields support for a set of predictions about the influence of possible selves on current behavior. In the other study, we examine possible selves in the context of an interpersonal model of self-regulation, showing strong evidence of concern for relational value in freely generated hoped-for and feared selves. These findings suggest that the role of possible selves in motivation and behavior can be profitably studied in models that fully specify the process of self-regulation and that those models can be enriched by a consideration of future-oriented self-representations. We offer additional recommendations for strengthening research on possible selves and self-regulation. PMID- 17083663 TI - Maintaining activity engagement: individual differences in the process of self regulating motivation. AB - Typically, models of self-regulation include motivation in terms of goals. Motivation is proposed to differ among individuals as a consequence of the goals they hold as well as how much they value those goals and expect to attain them. We suggest that goal-defined motivation is only one source of motivation critical for sustained engagement. A second source is the motivation that arises from the degree of interest experienced in the process of goal pursuit. Our model integrates both sources of motivation within the goal-striving process and suggests that individuals may actively monitor and regulate them. Conceptualizing motivation in terms of a self-regulatory process provides an organizing framework for understanding how individuals might differ in whether they experience interest while working toward goals, whether they persist without interest, and whether and how they try to create interest. We first present the self-regulation of motivation model and then review research illustrating how the consideration of individual differences at different points in the process allows a better understanding of variability in people's choices, efforts, and persistence over time. PMID- 17083664 TI - Self-regulation processes and health: the importance of optimism and goal adjustment. AB - This article discusses how self-regulatory models can be used to understand people's response to health threats. The article begins with a general discussion of the principles and assumptions of self-regulatory models of behavior. Two distinct lines of research are then presented addressing two important processes of adaptive self-regulation. First, we provide a brief overview of the literature on optimism and adjustment to chronic disease and other health outcomes. Second, we present an overview of the process of disengagement from unattainable goals, focusing on recent research. We close by making recommendations for future research. PMID- 17083665 TI - The pursuit of self-esteem: contingencies of self-worth and self-regulation. AB - Successful self-regulation is defined as the willingness to exert effort toward one's most important goals, while taking setbacks and failures as opportunities to learn, identify weaknesses and address them, and develop new strategies toward achieving those goals. Contingencies of self-worth can facilitate self-regulation because people are highly motivated to succeed and avoid failure in domains of contingency. However, because boosts in self-esteem are pleasurable and drops in self-esteem are painful, protection, maintenance, and enhancement of self-esteem can become the overriding goal. Several pitfalls for self-regulation can result, especially when tasks are difficult and failure is likely. In this article, we describe a program of research examining these self-regulation pitfalls associated with contingent self-worth and suggest that learning orientations, particularly the willingness to embrace failure for the learning it affords, foster successful self-regulation even in people with highly contingent self esteem. PMID- 17083666 TI - Self-regulation and personality: how interventions increase regulatory success, and how depletion moderates the effects of traits on behavior. AB - Self-regulation is a highly adaptive, distinctively human trait that enables people to override and alter their responses, including changing themselves so as to live up to social and other standards. Recent evidence indicates that self regulation often consumes a limited resource, akin to energy or strength, thereby creating a temporary state of ego depletion. This article summarizes recent evidence indicating that regular exercises in self-regulation can produce broad improvements in self-regulation (like strengthening a muscle), making people less vulnerable to ego depletion. Furthermore, it shows that ego depletion moderates the effects of many traits on behavior, particularly such that wide differences in socially disapproved motivations produce greater differences in behavior when ego depletion weakens the customary inner restraints. PMID- 17083667 TI - Hypo-egoic self-regulation: exercising self-control by diminishing the influence of the self. AB - Theory and research dealing with self-regulation have focused primarily on instances of self-regulation that involve high levels of self-reflection and effortful self-control. However, intentionally trying to control one's behavior sometimes reduces the likelihood of achieving one's goals. This article examines the process of hypo-egoic self-regulation in which people relinquish deliberate, conscious control over their own behavior so that they will respond more naturally, spontaneously, or automatically. An examination of spontaneously occurring hypo-egoic states (such as flow, deindividuation, and transcendence) suggests that hypo-egoic states are characterized by lowered self-awareness and/or an increase in concrete and present-focused self-thoughts. In light of this, people may intentionally foster hypo-egoism via two pathways-(a) taking steps to reduce the proportion of time that they are self-aware (such as repeating a behavior until it is automatic or practicing meditation) or (b) increasing the concreteness of their self-thoughts (such as inducing a concrete mindset or practicing mindfulness). In this way, people may deliberately choose to regulate hypo-egoically when effortful control might be detrimental to their performance. PMID- 17083668 TI - Mitochondrial genes leave home. PMID- 17083669 TI - The sweet stench of decay. PMID- 17083670 TI - Ferns represent an untapped biodiversity for improving crops for environmental stress tolerance. PMID- 17083672 TI - Plant CO2 responses: an issue of definition, time and resource supply. AB - In this review I am drawing attention to some constraints and biases in CO2 enrichment experiments and the analysis of data in the literature. Conclusions drawn from experimental works differ when the data are grouped in a way such that the relative frequency of test conditions does not determine the emerging trends, for instance unrealistically strong CO2-'fertilization' effects, which are in conflict with some basic ecological principles. I suggest separating three test conditions: uncoupled systems (plants not depending in a natural nutrient cycle) (I); expanding systems, in which plants are given ample space and time to explore otherwise limited resources (II); and fully coupled systems in which the natural nutrient cycling governs growth at steady-state leaf area index (LAI) and fine root renewal (III). Data for 10 type III experiments yield rather moderate effects of elevated CO2 on plant biomass production, if any. In steady-state grassland, the effects are water-related; in closed tree stands, initial effects decline rapidly with time. Plant-soil coupling (soil conditions) deserves far greater attention than plant-atmosphere coupling (CO2 enrichment technology). PMID- 17083673 TI - The evolution of plant-insect mutualisms. AB - Mutualisms (cooperative interactions between species) have had a central role in the generation and maintenance of life on earth. Insects and plants are involved in diverse forms of mutualism. Here we review evolutionary features of three prominent insect-plant mutualisms: pollination, protection and seed dispersal. We focus on addressing five central phenomena: evolutionary origins and maintenance of mutualism; the evolution of mutualistic traits; the evolution of specialization and generalization; coevolutionary processes; and the existence of cheating. Several features uniting very diverse insect-plant mutualisms are identified and their evolutionary implications are discussed: the involvement of one mobile and one sedentary partner; natural selection on plant rewards; the existence of a continuum from specialization to generalization; and the ubiquity of cheating, particularly on the part of insects. Plant-insect mutualisms have apparently both arisen and been lost repeatedly. Many adaptive hypotheses have been proposed to explain these transitions, and it is unlikely that any one of them dominates across interactions differing so widely in natural history. Evolutionary theory has a potentially important, but as yet largely unfilled, role to play in explaining the origins, maintenance, breakdown and evolution of insect-plant mutualisms. PMID- 17083674 TI - Evolutionary transfers of mitochondrial genes to the nucleus in the Populus lineage and coexpression of nuclear and mitochondrial Sdh4 genes. AB - The transfer of mitochondrial genes to the nucleus is an ongoing evolutionary process in flowering plants. Evolutionarily recent gene transfers provide insights into the evolutionary dynamics of the process and the way in which transferred genes become functional in the nucleus. Genes that are present in the mitochondrion of some angiosperms but have been transferred to the nucleus in the Populus lineage were identified by searches of Populus sequence databases. Sequence analyses and expression experiments were used to characterize the transferred genes. Two succinate dehydrogenase genes and six mitochondrial ribosomal protein genes have been transferred to the nucleus in the Populus lineage and have become expressed. Three transferred genes have gained an N terminal mitochondrial targeting presequence from other pre-existing genes and two of the transferred genes do not contain an N-terminal targeting presequence. Intact copies of the succinate dehydrogenase gene Sdh4 are present in both the mitochondrion and the nucleus. Both copies of Sdh4 are expressed in multiple organs of two Populus species and RNA editing occurs in the mitochondrial copy. These results provide a genome-wide perspective on mitochondrial genes that were transferred to the nucleus and became expressed, functional genes during the evolutionary history of Populus. PMID- 17083675 TI - Regulation of oil accumulation in single glands of Eucalyptus polybractea. AB - The accumulation of terpenoid oil was examined in the leaves of Eucalyptus polybractea at scales ranging from individual oil glands to the whole plant. Variations in oil composition and concentration of oil were measured and related to both morphological and physiological parameters. Within a plant, all glands produced oil of broadly similar composition that was not regulated by leaf age or the position of the gland within the leaf. There were, however, distinct differences between plants, suggesting that composition is controlled primarily at the whole-plant level. Oil concentration, too, was regulated primarily at the whole-plant level and was limited by gland capacity. Gland capacity was linked to leaf area and thickness, the final products of leaf expansion. Leaf and plant oil composition is determined not by a mosaic of glands specializing in producing a single or a small group of compounds, but rather by glands with remarkably similar capacities for terpenoid biosynthesis, although oil concentration, limited by gland capacity, may be linked to leaf expansion rather than biosynthetic capacity. PMID- 17083676 TI - The chemical nature of fetid floral odours in stapeliads (Apocynaceae Asclepiadoideae-Ceropegieae). AB - By emitting strong fetid scents, sapromyiophilous flowers mimic brood and food sites of flies to attract them as pollinators. To date, intensive comparative scent analyses have been restricted to sapromyiophilous Araceae. Here, we analysed flower volatiles of fetid stapeliads to improve our understanding of the floral biology of fly pollinated species, and to learn whether mimicry types comparable to those found in Araceae exist. Floral volatiles of 15 species out of 11 genera within the Asclepiadoideae-Ceropegieae-Stapeliinae were collected via headspace adsorption and thermal desorption and analysed by gas chromatography mass spectometry (GC-MS). Data were analysed using CNESS-NMDS statistics. Sapromyiophilous stapeliads are highly diverse in their scent composition, in which sulphur compounds, benzenoids, fatty acid derivatives or nitrogen containing compounds dominate. Four groups are evident: species with high p cresol content but low amounts of polysulphides (herbivore faeces mimicry); species with mainly polysulphides and low amounts of p-cresol (carnivore/omnivore faeces or carcass mimicry); species with high amounts of heptanal and octanal (carnivore/omnivore faeces or carcass mimicry); and species with hexanoic acid (urine mimicry). Considering the findings in the unrelated Araceae, our results support the universality of different mimicry types that are obviously subsumed under the sapromyiophilous syndrome. PMID- 17083677 TI - Mercury hinders recovery of shoot hydraulic conductivity during grapevine rehydration: evidence from a whole-plant approach. AB - This experiment aimed to test whether recovery of shoot hydraulic conductivity after drought depends on cellular metabolism in addition to xylem hydraulics. We rehydrated droughted grapevines (Vitis vinifera) after treating intact plants through the root with 0.5 mm mercuric chloride (a metabolic inhibitor) at the end of the stress period, before rehydration. The contribution of mercury-inhibited water transport in both shoot and root, and the extent of shoot vessel embolization, were assessed. Drought stress decreased plant water potential and induced embolization of the shoot vessels. The rehydration in Hg-untreated plants re-established both shoot water potential and specific shoot hydraulic conductivity (Kss) at levels comparable with watered controls, and induced recovery of most of the embolisms formed in the shoot during the drought. In contrast, in plants treated with HgCl2, recovery of Kss and root hydraulic conductance were impaired. In rehydrated, Hg-treated plants, the effects of Hg on Kss were reversed when either the shoot or the root was treated with 60 mM beta mercaptoethanol as a mercuric scavenger. This work suggests that plant cellular metabolism, sensitive to mercuric chloride, affects the recovery of shoot hydraulic conductivity during grapevine rehydration by interfering with embolism removal, and that it involves either the root or the shoot level. PMID- 17083678 TI - Correlated evolution of stem and leaf hydraulic traits in Pereskia (Cactaceae). AB - Recent studies have demonstrated significant correlations between stem and leaf hydraulic properties when comparing across species within ecological communities. This implies that these traits are co-evolving, but there have been few studies addressing plant water relations within an explicitly evolutionary framework. This study tests for correlated evolution among a suite of plant water-use traits and environmental parameters in seven species of Pereskia (Cactaceae), using phylogenetically independent contrasts. There were significant evolutionary correlations between leaf-specific xylem hydraulic conductivity, Huber Value, leaf stomatal pore index, leaf venation density and leaf size, but none of these traits appeared to be correlated with environmental water availability; only two water relations traits - mid-day leaf water potentials and photosynthetic water use efficiency - correlated with estimates of moisture regime. In Pereskia, it appears that many stem and leaf hydraulic properties thought to be critical to whole-plant water use have not evolved in response to habitat shifts in water availability. This may be because of the extremely conservative stomatal behavior and particular rooting strategy demonstrated by all Pereskia species investigated. These results highlight the need for a lineage-based approach to understand the relative roles of functional traits in ecological adaptation. PMID- 17083679 TI - Hydrogen exchange during cellulose synthesis distinguishes climatic and biochemical isotope fractionations in tree rings. AB - The abundance of the hydrogen isotope deuterium (D) in tree rings is an attractive record of climate; however, use of this record has proved difficult so far, presumably because climatic and physiological influences on D abundance are difficult to distinguish. Using D labelling, we created a D gradient in trees. Leaf soluble sugars of relatively low D abundance entered cellulose synthesis in stems containing strongly D-labelled water. We used nuclear magnetic resonance (NMR) spectroscopy to quantify D in the C-H groups of leaf glucose and of tree ring cellulose. Ratios of D abundances of individual C-H groups of leaf glucose depended only weakly on leaf D labelling, indicating that the D abundance pattern was determined by physiological influences. The D abundance pattern of tree-ring cellulose revealed C-H groups that exchanged strongly (C(2)-H) or weakly (C(6) H2) with water during cellulose synthesis. We propose that strongly exchanging C H groups of tree-ring cellulose adopt a climate signal stemming from the D abundance of source water. C-H groups that exchange weakly retain their D abundance established in leaf glucose, which reflects physiological influences. Combining both types of groups may allow simultaneous reconstruction of climate and physiology from tree rings. PMID- 17083680 TI - Leaf carbohydrate controls over Arabidopsis growth and response to elevated CO2: an experimentally based model. AB - Transient starch production is thought to strongly control plant growth and response to elevated CO2. We tested this hypothesis with an experimentally based mechanistic model in Arabidopsis thaliana. Experiments were conducted on wild type (WT) A. thaliana, starch-excess (sex1) and starchless (pgm) mutants under ambient and elevated CO2 conditions to determine parameters and validate the model. The model correctly predicted that mutant growth is approx. 20% of that in WT, and the absolute response of both mutants to elevated CO2 is an order of magnitude lower than in WT. For sex1, direct starch unavailability explained the growth responses. For pgm, we demonstrated experimentally that maintenance respiration is proportional to leaf soluble sugar concentration, which gave the necessary feedback mechanism on modelled growth. Our study suggests that the effects of sugar-starch cycling on growth can be explained by simple allocation processes, and the maximum rate of leaf growth (sink capacity) exerts a strong control over the response to elevated CO2 of herbaceous plants such as A. thaliana. PMID- 17083681 TI - Differences in plant function in phosphorus- and nitrogen-limited mangrove ecosystems. AB - Mangrove ecosystems can be either nitrogen (N) or phosphorus (P) limited and are therefore vulnerable to nutrient pollution. Nutrient enrichment with either N or P may have differing effects on ecosystems because of underlying differences in plant physiological responses to these nutrients in either N- or P-limited settings. Using a common mangrove species, Avicennia germinans, in sites where growth was either N or P limited, we investigated differing physiological responses to N and P limitation and fertilization. We tested the hypothesis that water uptake and transport, and hydraulic architecture, were the main processes limiting productivity at the P-limited site, but that this was not the case at the N-limited site. We found that plants at the P-deficient site had lower leaf water potential, stomatal conductance and photosynthetic carbon-assimilation rates, and less conductive xylem, than those at the N-limited site. These differences were greatly reduced with P fertilization at the P-limited site. By contrast, fertilization with N at the N-limited site had little effect on either photosynthetic or hydraulic traits. We conclude that growth in N- and P-limited sites differentially affect the hydraulic pathways of mangroves. Plants experiencing P limitation appear to be water deficient and undergo more pronounced changes in structure and function with relief of nutrient deficiency than those in N-limited ecosystems. PMID- 17083682 TI - Fine-root turnover patterns and their relationship to root diameter and soil depth in a 14C-labeled hardwood forest. AB - Characterization of turnover times of fine roots is essential to understanding patterns of carbon allocation in plants and describing forest C cycling. We used the rate of decline in the ratio of 14C to 12C in a mature hardwood forest, enriched by an inadvertent 14C pulse, to investigate fine-root turnover and its relationship with fine-root diameter and soil depth. Biomass and Delta14C values were determined for fine roots collected during three consecutive winters from four sites, by depth, diameter size classes (< 0.5 or 0.5-2 mm), and live-or-dead status. Live-root pools retained significant 14C enrichment over 3 yr, demonstrating a mean turnover time on the order of years. However, elevated Delta14C values in dead-root pools within 18 months of the pulse indicated an additional component of live roots with short turnover times (months). Our results challenge assumptions of a single live fine-root pool with a unimodal and normal age distribution. Live fine roots < 0.5 mm and those near the surface, especially those in the O horizon, had more rapid turnover than 0.5-2 mm roots and deeper roots, respectively. PMID- 17083683 TI - Arbuscular mycorrhizal fungi contribute to phosphorus uptake by wheat grown in a phosphorus-fixing soil even in the absence of positive growth responses. AB - We used 32P to quantify the contribution of an arbuscular mycorrhizal (AM) fungus (Glomus intraradices) to phosphorus (P) uptake by wheat (Triticum aestivum), grown in compartmented pots. The soil was from a major cereal-growing area, the Eyre Peninsula, South Australia; it was highly calcareous and P-fixing. Fertilizer P was added to soil at 20 mg kg(-1), as solid or liquid. Two extraction methods were used to estimate plant-available P. Fungal colonization was well established at harvest (36 d). Application of P decreased both colonization and hyphal length density in soil, with small differences between different P fertilizers. Plants showed large positive responses in terms of growth or total P uptake to all P additions, and showed no positive (or even negative) responses to AM colonization, regardless of P application. 32P was detected only in AM plants, and we calculated that over 50% of P uptake by plants was absorbed via AM fungi, even when P was added. The results add to the growing body of knowledge that 'nonresponsive' AM plants have a functional AM pathway for P transfer to the plant; it should not be ignored in breeding plants for root traits designed to improve P uptake. PMID- 17083684 TI - Arbuscular mycorrhizal colonization on carbon economy in perennial ryegrass: quantification by 13CO2/12CO2 steady-state labelling and gas exchange. AB - Effects of the arbuscular mycorrhizal fungus (AMF) Glomus hoi on the carbon economy of perennial ryegrass (Lolium perenne) were investigated by comparing nonmycorrhizal and mycorrhizal plants of the same size, morphology and phosphorus status. Plants were grown in the presence of CO2 sources with different C isotope composition (delta13C -1 or -44). Relative respiration and gross photosynthesis rates, and belowground allocation of C assimilated during one light period ('new C'), as well as its contribution to respiration, were quantified by the concerted use of 13CO2/12CO2 steady-state labelling and 13CO2/12CO2 gas-exchange techniques. AMF (G. hoi) enhanced the relative respiration rate of the root + soil system by 16%, inducing an extra C flow amounting to 3% of daily gross photosynthesis. Total C flow into AMF growth and respiration was estimated at < 8% of daily gross photosynthesis. This was associated with a greater amount of new C allocated belowground and respired in mycorrhizal plants. AMF colonization affected the sources supplying belowground respiration, indicating a greater importance of plant C stores in supplying respiration and/or the participation of storage pools within fungal tissues. When ontogenetic and nutritional effects were accounted for, AMF increased belowground C costs, which were not compensated by increased photosynthesis rates. Therefore the instantaneous relative growth rate was lower in mycorrhizal plants. PMID- 17083685 TI - Mycorrhizal fungal identity and richness determine the diversity and productivity of a tallgrass prairie system. AB - We investigated the effects of arbuscular mycorrhizal fungal (AMF) species richness and composition on plant community productivity and diversity, and whether AMF mediate plant species coexistence by promoting niche differentiation in phosphorus use. Our experiment manipulated AMF species richness and identity across a range of P conditions in tallgrass prairie mesocosms. We showed that increasing AMF richness promoted plant diversity and productivity, but that this AMF richness effect was small relative to the effects of individual AMF species. We found little support for AMF-facilitated complementarity in P use. Rather, the AMF richness effect appeared to be caused by the inclusion of particular diversity- and productivity-promoting AMF (a sampling effect). Furthermore, the identity of the diversity-promoting fungi changed with P environment, as did the relationship between the diversity-promoting and productivity-promoting benefits of AMF. Our results suggest that plant diversity and productivity are more responsive to AMF identity than to AMF diversity per se, and that AMF identity and P environment can interact in complex ways to alter community-level properties. PMID- 17083686 TI - Multivesicular compartments proliferate in susceptible and resistant MLA12-barley leaves in response to infection by the biotrophic powdery mildew fungus. AB - There is growing evidence that multivesicular bodies and cell wall-associated paramural bodies participate in the enhanced vesicle trafficking induced by pathogen attack. Here, we performed transmission electron microscopy in combination with cytochemical localization of H2O2 to investigate multivesicular compartments during establishment of compatible interaction in susceptible barley (Hordeum vulgare) and during hypersensitive response in resistant MLA12-barley infected by the barley powdery mildew fungus (Blumeria graminis f. sp. hordei). Multivesicular bodies, intravacuolar vesicle aggregates and paramural bodies proliferated in the penetrated epidermal cell during development of the fungal haustorium. These vesicular structures also proliferated at the periphery of intact cells, which were adjacent to the hypersensitive dying cells and deposited cell wall appositions associated with H2O2 accumulation. All plasmodesmata between intact cells and hypersensitive cells were constricted or blocked by cell wall appositions. These results suggest that multivesicular compartments participate in secretion of building blocks for cell wall appositions not only to arrest fungal penetration but also to contain hypersensitive cell death through blocking plasmodesmata. They may also participate in internalization of damaged membranes, deleterious materials, nutrients, elicitors and elicitor receptors. PMID- 17083687 TI - Variability patterns and positively selected sites at the gametophytic self incompatibility pollen SFB gene in a wild self-incompatible Prunus spinosa (Rosaceae) population. AB - Current models for the generation of new gametophytic self-incompatibility specificities require that neutral variability segregates within specificity classes. Furthermore, one of the models predicts greater ratios of nonsynonymous to synonymous substitutions in pollen than in pistil specificity genes. All models assume that new specificities arise by mutation only. To test these models, 21 SFB (the pollen S-locus) alleles from a wild Prunus spinosa (Rosaceae) population were obtained. For seven of these, the corresponding S-haplotype was also characterized. The SFB data set was also used to identify positively selected sites. Those sites are likely to be the ones responsible for defining pollen specificities. Of the 23 sites identified as being positively selected, 21 are located in the variable (including a new region described here) and hypervariable regions. Little variability is found within specificity classes. There is no evidence for selective sweeps being more frequent in pollen than in pistil specificity genes. The S-RNase and the SFB genes have only partially correlated evolutionary histories. None of the models is compatible with the variability patterns found in the SFB and the S-haplotype data. PMID- 17083688 TI - Detection of clonal rearrangement of T-cell receptor genes in the diagnosis of primary cutaneous CD30 lymphoproliferative disorders. AB - BACKGROUND: Detection of clonality has been reported to be a helpful tool in the diagnosis of cutaneous lymphomas. Monoclonal rearrangement of T-cell receptor genes (TCR) was reported in fresh frozen tissue of lymphomatoid papulosis (LyP) and primary cutaneous anaplastic large-cell lymphoma (ALCL), but the diagnostic value of T-cell clonality in formalin-fixed, paraffin-embedded biopsies has so far not been assessed. METHODS: Detection of clonal rearrangement of TCRgamma genes by highly sensitive polymerase chain reaction-based automated high resolution fragment analysis (AHRFA) in archival LyP (n = 18) and ALCL (n = 17) tissue. RESULTS: Detection of clonality differed significantly among the histologic forms of LyP as well as between LyP and ALCL with clonality found in none of the 10 biopsies of LyP type A and B, in 4/8 (50%) of the LyP type C specimens, and in 11/17 (65%) of ALCL cases. CONCLUSIONS: T-cell clonality can only be found in a minority (four of 18; 22%) of archival LyP specimens, even when employing a highly sensitive detection method and is thus of limited diagnostic value. Final diagnosis of LyP has to be based mainly on clinical, histologic, and immunohistochemical findings rather than on results of clonality studies. PMID- 17083689 TI - Tenascin expression in actinic keratosis. AB - BACKGROUND: Tenascin is an extracellular matrix protein frequently expressed around neoplastic and non-neoplastic lesions of the skin. Actinic keratoses (AKs) are intraepidermal neoplastic lesions of the sun-exposed skin. They are classified according to the extension of dysplasia in four stages; they also present different histological varieties. METHODS: We performed an immunohistochemical study using tenascin monoclonal antibody diluted 1 : 50 on 150 cases of AKs classified, respectively, in histotypes (38 hypertrophic, 18 atrophic, 21 bowenoid, 19 acantolytic, and 40 mixed) and in stages (27 stage I, 46 stage II, 42 stage III, and 35 stage IV; 14 in tumoral progression). RESULTS: Tenascin positivity was observed in all cases at the dermal level close to the epithelial lesion. The intensity of reaction increased from stage I to stage IV and, of course, also in tumoral progression. Its expression was not related to the histotypes. In very few cases, the atypical keratinocytes were positive. CONCLUSIONS: Tenascin expression in AKs is related to the stages of dysplasia. In fact, the immunostaining intensity corresponds to the degree of the dysplasia rather than the thickness of the involved epidermis. Tenascin plays a role in neoplastic progression working as an anti-adhesive factor. PMID- 17083690 TI - Non-bullous neutrophilic dermatosis: an uncommon dermatologic manifestation in patients with lupus erythematosus. AB - BACKGROUND: Rare cases of a non-bullous neutrophilic dermatosis occurring in patients with lupus erythematosus (LE) have been reported, often as the presenting manifestation of the disease. METHODS: We reviewed hematoxylin and eosin-stained slides and obtained clinical information from four additional patients, two of whom had no prior history of LE. RESULTS: All patients were female, aged 16-59 (mean age 37). Clinically, the skin lesions were characterized by widely distributed pruritic papules and plaques. Three patients presented with systemic symptoms, including fever, arthritis and malaise. Histopathologic examination in all cases showed a superficial perivascular and interstitial neutrophilic infiltrate with leukocytoclasis. There was no evidence of vasculitis. Mild focal vacuolar change was a subtle feature seen only in the biopsies of the two patients with a prior history of LE. CONCLUSIONS: It is important to consider LE in the differential diagnosis of non-bullous neutrophilic dermatoses. PMID- 17083691 TI - Hidradenocarcinoma: a histological and immunohistochemical study. AB - The diagnosis of hidradenocarcinoma is difficult due to a combination of factors including inconsistent nomenclature/ classification, rarity of the neoplasm, and variable morphology of cells composing the neoplasm. Immunohistochemistry has not been previously performed on a series of hidradenocarcinomas. We evaluated six cases of hidradenocarcinoma histologically and immunohistochemically using antibodies to gross cystic disease fluid protein-15 (GCDFP-15), carcino-embryonic antigen (CEA), epithelial membrane antigen (EMA), S-100 protein, keratin AE1/3, cytokeratin 5/6, p53, bcl-1, bcl-2, and Ki67. Histology suggested concurrent eccrine and apocrine differentiation of the cases. Ki67 and p53 staining was strongly positive in five of six tumors. The neoplasms stained with antibodies to CEA, S-100 protein, GCDFP-15, EMA, bcl-1, and bcl-2 in no consistent pattern. All tumors studied stained positively for keratin AE1/3 and cytokeratin 5/6. In making the diagnosis of hidradenocarcinoma, it may be unnecessary to separate hidradenocarcinoma into eccrine and apocrine categories, and although Ki67 and p53 may be helpful, histological parameters remain paramount. PMID- 17083692 TI - Narrow-spectrum staining pattern of Pityrosporum. AB - BACKGROUND: Broad-spectrum fungal stains are used to detect fungal organisms, but narrow-spectrum stains can assist in fungal differential diagnosis. These stains include mucicarmine and Fontana-Masson (FM) for Cryptococcus, Alcian Blue for Cryptococcus and Blastomyces, Congo Red for Blastomyces and Coccidioides, and Ziehl-Neelsen for some examples of Blastomyces and Histoplasma. Pityrosporum is increasingly being recognized as a pathogen capable of significant cutaneous and systemic infections, but the narrow-spectrum staining pattern of Pityrosporum has not yet been systematically studied and reported. METHODS: Paraffin-embedded archival tissue from 10 skin biopsies containing Pityrosporum was stained with Alcian Blue, Congo Red, Ziehl-Neelsen acid-fast stain, and FM stain. In addition to the 60-min staining time, a modified FM stain with 15-, 30-, and 45-min staining times was also performed. RESULTS: All cases stained positive with Alcian Blue, Congo Red, and FM stains and negative with the Ziehl-Neelsen stain. The modified FM stain with the 15-min staining time showed significant staining in more than 50% of the cases. DISCUSSION: The narrow-spectrum staining pattern of Pityrosporum distinguishes it from Cryptococcus, Blastomyces, Candida, Histoplasma, and Coccidioides. PMID- 17083693 TI - Symplastic hemangioma: report of two cases. AB - Symplastic hemangioma is characterized by degenerative atypia of vascular smooth muscle and interstitial cells within a pre-existing vascular lesion with minimal endothelial cell atypia. We describe an additional two cases of this distinctive but poorly recognized entity. On histology, both lesions revealed a cirsoid aneurysm-type appearance with thick-walled and variably dilated blood vessels. The vascular endothelial cells showed mild nuclear hyperchromasia with no multilayering or mitoses. The atypical cells, either located within the vascular smooth muscle wall or within the interstitium, were spindle or epithelioid with varying degrees of hyperchromasia, nuclear enlargement, pleomorphism, and multinucleation. Perivascular hemorrhage, vascular thrombosis, and focal papillary endothelial hyperplasia were uniformly present. The variably fibrous to edematous stroma showed hemosiderin deposits and a mononuclear inflammatory infiltrate. Clusters of adipocytes were present within the superficial dermis. Rare atypical mitoses and occasional bizarre lipoblast-like stromal cells were identified in one tumor. Immunohistochemistry showed focal smooth muscle actin positivity in the pleomorphic cells of the vascular walls. CD68 and CD34 stained occasional stromal cells in the interstitial location. Both the cases showed no recurrence. The bizarre cytologic changes are interpreted as degenerative in nature and probably akin to that observed in ancient schwannoma and uterine symplastic leiomyoma. PMID- 17083694 TI - Female-patterned alopecia in teenage brothers with unusual histologic features. AB - BACKGROUND: Patterned hair loss, follicular miniaturization, and increased telogen hair counts characterize androgenic alopecia (AGA). Follicular inflammation in AGA has been associated with treatment resistance and progressive hair loss. CASE REPORT: Brothers, 15 and 18 years old, presented with frontal and mid-scalp hair loss with an intact frontal hairline noted over a 1-year period. The elder reported past use of androgenic steroids. Laboratory assessment for metabolic and hormonal abnormalities was unrevealing, and hair pull test was negative. Scalp biopsies revealed decreased terminal hairs, marked diameter variation of anagen hairs, decreased terminal to vellus hair ratios (3.7:1/3.4:1, older/younger), and increased telogen counts (23%/21%). Infrabulbar and peri isthmic (follicular bulge region) lymphocytic infiltrates were present. Hair loss has progressed, unabated by daily topical 0.5% clobetasol (for 6 months), daily 5% minoxidil (1 year), and latter, daily oral finasteride (2 years - older brother only). DISCUSSION: Based on patterned hair loss and miniaturized hairs, these brothers have AGA. The female pattern of hair loss (diffuse hair loss affecting the central scalp with preservation of frontal hair line) coupled with follicular isthmic lymphocytic inflammation represents an unusual presentation, possibly a treatment resistant, inflammatory variant of AGA. The differential diagnosis includes exogenous androgen-mediated hair loss, cicatricial pattern hair loss, or the superimposition of alopecia areata. PMID- 17083695 TI - Subcutaneous ossifying fibromyxoid tumor. AB - Ossifying fibromyxoid tumor (OFMT) is an uncommon neoplasm occurring predominantly in the deep soft tissues of the trunk and proximal extremities. The lineage of this rare tumor to date is still uncertain. We present a case of a subcutaneous OFMT that recurred 8 years after initial resection. The histological findings of the primary and recurrent lesions are compared along with the histologic features possibly associated with aggressiveness. Dermatopathologists should consider OFMT in the differential diagnosis of subcutaneous neoplasms with a myxoid matrix. PMID- 17083696 TI - Skin metastasis of 'nested type' of urothelial carcinoma of the urinary bladder. PMID- 17083697 TI - Can scleromyxedema be differentiated from nephrogenic fibrosing dermatopathy by the distribution of the infiltrate? PMID- 17083698 TI - Fibrolamellar nerve sheath tumor or sclerotic neurofibroma? PMID- 17083699 TI - Cellular digital fibromas: what about superficial acral fibromyxoma? PMID- 17083701 TI - Diagnosis of American foulbrood in honey bees: a synthesis and proposed analytical protocols. AB - Worldwide, American foulbrood (AFB) is the most devastating bacterial disease of the honey bee (Apis mellifera). Because the distinction between AFB and powdery scale disease is no longer considered valid, the pathogenic agent has recently been reclassified as one species Paenibacillus larvae, eliminating the subspecies designations Paenibacillus larvae subsp. larvae and Paenibacillus larvae subsp. pulvifaciens. The creamy or dark brown, glue-like larval remains of infected larvae continue to provide the most obvious clinical symptom of AFB, although it is not conclusive. Several sensitive and selective culture media are available for isolation of this spore-forming bacterium, with the type of samples that may be utilized for detection of the organism being further expanded. PCR methods for identification and genotyping of the pathogen have now been extensively developed. Nevertheless, biochemical profiling, bacteriophage sensitivity, immunotechniques and microscopy of suspect bacterial strains are entirely adequate for routine identification purposes. PMID- 17083702 TI - Campylobacter in chicken livers and their destruction by pan frying. AB - AIM: To enumerate Campylobacter spp. on the external surface and internal portions of chicken livers, and to assess the cooking required to inactivate naturally present cells. METHODS AND RESULTS: Of 30 livers tested all yielded Campylobacter spp. on their surfaces and 90% were found to contain the organism in internal tissue. Four (13%) livers contained >10(4) MPN campylobacters, and an additional seven (23%) contained >10(3) MPN campylobacters per liver. The internal temperature of pan-fried livers under the conditions used reached a maximum of 70-80 degrees C, and maintaining this temperature for 2-3 min was necessary to inactivate naturally occurring Campylobacter spp. All isolates identified were either C. jejuni or C. coli. CONCLUSIONS: Chicken livers represent a potential source of human campylobacteriosis as they contained >10(4) MPN per liver in 13% of the samples tested. Pan-frying can produce an acceptable product that is safe to eat. SIGNIFICANCE AND IMPACT OF THIS STUDY: The data provided can be used in exposure assessments of Campylobacter in poultry products in terms of both quantitative data and assessing pan-frying and its ability to destroy campylobacters. PMID- 17083703 TI - Growth temperature of four Campylobacter jejuni strains influences their subsequent survival in food and water. AB - AIM: To determine if Campylobacter jejuni grown at 37 and 42 degrees C have different abilities to survive on beef and chicken, and in water. METHODS AND RESULTS: Beef, chicken and water were separately inoculated with four Camp. jejuni (two poultry and two beef) strains grown at 37 or 42 degrees C. The matrices were stored at approximately 4 degrees C and Camp. jejuni numbers were monitored over time by plate counts. On beef there was a greater decrease in number for two strains (P < 0.05; approximately 0.7 and 1.3 log CFU cm(-2)) grown at 37 degrees C as compared with 42 degrees C. By contrast on chicken there was a decrease in numbers for two strains (P < 0.05; approximately 1.3 and 1 log CFU g( 1)) grown at 42 degrees C as compared with 37 degrees C. In water there was a greater decrease in numbers for all strains (P < 0.05; approximately 3-5.3 log CFU ml(-1)) grown at 42 degrees C as compared with 37 degrees C. CONCLUSIONS: Growth temperature influences the survival of Camp. jejuni on food and in water. SIGNIFICANCE AND IMPACT OF THE STUDY: Campylobacter jejuni survival studies need to consider growth temperature to avoid erroneous results. Campylobacter jejuni grown at 37 degrees C, the body temperature of humans and cattle, may represent a greater public health risk in water than those grown at 42 degrees C, the body temperature of poultry. PMID- 17083704 TI - Methicillin-resistant staphylococcal colonization in clinically normal dogs and horses in the community. AB - AIMS: To evaluate the prevalence of methicillin-resistant staphylococcal (MRS) colonization in clinically normal dogs and horses in the community. METHODS AND RESULTS: Three hundred clinically normal horses and 200 clinically normal dogs were enrolled. One nasal swab was collected from each horse. Two swabs were taken from each dog: (i) from an anterior nare, and (ii) a combination of the perineal area and 0.5 cm into the anus. Enrichment cultures were performed. Methicillin resistant Staphylococcus aureus (MRSA) was not identified. Methicillin-resistant Staphylococcus intermedius (MRSI) was isolated from the nasal swab from three dogs. Methicillin-resistant coagulase negative staphylococci (MRCoNS) were isolated from 126/300 (42%) horses and 26/200 (13%) dogs. CONCLUSIONS: At present MRSI is not considered to be a significant zoonotic concern; however, it may become an important pathogen in dogs. MRCoNS mostly cause disease in compromised human or animal hosts. However, these bacteria can serve as reservoirs of resistance determinants in the community, which could lead to the emergence of novel MRSA strains. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of the prevalence of MRS colonization in clinically normal dogs in a community setting. Continued surveillance is indicated to determine whether MRSA will emerge in the animal population and become a concern for animal disease and zoonotic infection. PMID- 17083705 TI - A study on medium chain length-polyhydroxyalkanoate accumulation in Escherichia coli harbouring phaC1 gene of indigenous Pseudomonas sp. LDC-5. AB - AIMS: This study is mainly focused on the heterologous expression and accumulation of polyhydroxyalkanoates (PHA) in Escherichia coli. METHODS AND RESULTS: PHA synthase gene (phaC1) from indigenous Pseudomonas sp. LDC-5 was amplified by PCR and cloned in E. coli (Qiagen EZ competent cells). The recombinant E. coli was analysed and confirmed for its expression of phaC1 gene by phase contrast microscopy, Western blot analysis and spectral studies (Fourier transform infrared spectroscopy). It was further evaluated for its accumulation in different carbon and nitrogen sources. The accumulation of PHA (3.4 g l(-1)) was enhanced in the medium supplemented with glycerol and fish peptone compared to the other carbon and nitrogen sources used in this study. CONCLUSIONS: This study would enable the reduction of cost of PHA production. SIGNIFICANCE AND IMPACT OF THE STUDY: An important part of this study is that E. coli harbouring partial phaC1 gene could accumulate medium chain length PHA significantly. The results demonstrated that the E. coli strain could be a potential candidate for the large-scale production of polymer. The conditions for the higher yield and productivity will be optimized in the next phase using fermentation studies. PMID- 17083706 TI - Evaluation of a new chromogenic agar medium for isolation and identification of Group B streptococci. AB - AIMS: To evaluate a new chromogenic agar as a screening medium for the isolation of Group B streptococci from high vaginal swabs from pregnant women. METHODS AND RESULTS: The medium was evaluated with 195 high vaginal swabs referred for antenatal screening and compared with blood agar and Granada medium. The new chromogenic medium showed 100% sensitivity for the detection of Group B streptococci, and also showed a positive predictive value of 100%. Granada medium also showed excellent sensitivity and specificity and both media were superior to blood agar. CONCLUSIONS: The new chromogenic medium showed excellent performance for the detection of Group B streptococci from clinical samples. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first chromogenic medium described for the detection of Group B streptococci. The medium offers an effective and convenient alternative to conventional media, currently used in clinical laboratories. PMID- 17083707 TI - Induced thermotolerance under nonisothermal treatments of a heat sensitive and a resistant strain of Staphylococcus aureus in media of different pH. AB - AIMS: The aim was to assess the induced thermotolerance under nonisothermal treatments of two strains of Staphylococcus aureus in media of different pH. METHODS AND RESULTS: Staphylococcus aureus ATCC 25923 was more heat resistant than S. aureus ATCC 13565 at any pH investigated under isothermal conditions. At pH 7.4, the D58 value of the resistant strain was approx. 30 times greater. Both strains showed a higher heat resistance at pH 4.0 than at pH 7.4. In contrast, under nonisothermal treatments (0.5-2 degrees C min(-1)), both strains were more heat resistant when treated at pH 7.4 than at pH 4.0 due to heat adaptation at the higher pH. At the slowest heating up rate tested at pH 7.4, the initially heat-sensitive strain nearly reached the thermotolerance of the heat-resistant strain. CONCLUSIONS: The induced thermotolerance under nonisothermal treatments depended on the treatment medium pH and the microbial strain tested. The induced thermotolerance in a sensitive strain can be greater than in a heat-resistant strain, showing similar resistance under nonisothermal conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: This work shows data of interest about mechanisms of microbial resistance and adaptation to heat. Moreover, it contributes to the development of more adequate combined processes for food preservation. PMID- 17083708 TI - Influence of growth conditions on the production of extracellular proteolytic enzymes in Paenibacillus peoriae NRRL BD-62 and Paenibacillus polymyxa SCE2. AB - AIMS: To analyse the extracellular protease profile of two Paenibacillus species, Paenibacillus peoriae and Paenibacillus polymyxa, as well as how different growth media influenced its expression. METHODS AND RESULTS: Both bacteria were cultured in five media [Luria-Bertani broth, glucose broth, thiamine/biotin/nitrogen broth (TBN), trypticase soy broth and a defined medium] for 48 h at 32 degrees C. Our results showed a heterogeneous protease secretion pattern whose expression was dependent on medium composition. However, TBN induced the most quantitative and qualitative protease production on both Paenibacillus. The proteases were detected in neutral-alkaline pH range, being totally inhibited by 1,10 phenanthroline, a zinc-metalloprotease inhibitor. We also analysed the protease expression during the growth and, at least to P. peoriae, the most elevated protease activity was measured at 96 h, in which the highest number of spores and a low concentration of viable cells were observed. CONCLUSIONS: The results presented add P. peoriae and P. polymyxa to the list of neutral-alkaline extracellular protease producers. SIGNIFICANCE AND IMPACT OF THE STUDY: Paenibacillus species are ubiquitous in nature, are capable to form resistant spores and to produce several hydrolytic enzymes, including proteases. However, only few data concerning the production of these enzymes are available. Proteases produced by Paenibacillus strains may represent new sources for biotechnological use. PMID- 17083709 TI - Simple detection of small amounts of Pseudomonas cells in milk by using a microfluidic device. AB - AIMS: Flow cytometry offers rapid and reliable analyses of bacteria in milk. However, a flow cytometer is relatively expensive and operation is rather complicated for an unskilled operator. We applied flow cytometry using a microfluidic device (on-chip flow cytometry) in detection of small amounts of milk-spoiling bacteria. METHODS AND RESULTS: Pseudomonas cells in milk were in situ hybridized with Cy5-labelled probe specific for Pseudomonas spp. under optimized condition. Numbers of Pseudomonas cells in the stationary phase and in the starved state determined by on-chip flow cytometry were compared with those determined by conventional plate counting, and on-chip flow cytometry detected targeted cells in milk that were undetectable as colony forming units(CFU) on Standards Methods Agar. CONCLUSIONS: The contamination in milk with fewer than 10 CFU ml(-1) of targeted cells in starved state was detectable with simple procedure (0.5 h milk-clearing, 1 h fixation, 2 h hybridization and 0.5 h on-chip flow cytometry following 12 h enrichment of cells). SIGNIFICANCE AND IMPACT OF THE STUDY: On-chip flow cytometry following fluorescence in situ hybridization could be applicable to simple detection of milk-spoiling bacteria. PMID- 17083710 TI - Discrepancies between the phenotypic and genotypic characterization of Lactococcus lactis cheese isolates. AB - AIMS: The use of randomly amplified polymorphic DNA (RAPD)-PCR fingerprinting and plasmid profiles to determine at the strain level, the similarity of Lactococcus lactis isolates obtained during sampling of traditional cheeses and to verify its correspondence to the selected phenotypic characteristics. METHODS AND RESULTS: A total of 45 L. lactis isolates were genotypically analysed by RAPD-PCR fingerprinting and plasmid patterns. Phenotypic traits used to compare strains were proteolytic, acidifying, aminotransferase (aromatic and branched chain aminotransferase) and alpha-ketoisovalerate decarboxylase (Kivd) activities. The results show that 23 isolates could be grouped in clusters that exhibited 100% identity in both their RAPD and plasmid patterns, indicating the probable isolation of dominant strains during the cheese sampling process. However, there were phenotypic differences between isolates within the same cluster that included the loss of relevant technological properties such as proteinase activity and acidifying capacity or high variation in their amino acid converting enzyme activities. Likewise, the analysis of a specific attribute, Kivd activity, indicated that 7 of 15 isolates showed no detectable activity despite the presence of the encoding (kivd) gene. CONCLUSION: Phenotypic differences found between genotypically similar strains of L. lactis strains could be linked to differences in enzymatic expression. SIGNIFICANCE AND IMPACT OF THE STUDY: Phenotypic analysis of L. lactis isolates should be considered when selecting strains with new cheese flavour forming capabilities. PMID- 17083711 TI - Norovirus binds to blood group A-like antigens in oyster gastrointestinal cells. AB - AIMS: To determine if histo-blood group antigens (HBGA) present in oyster gastrointestinal (GI) cells mediate accumulation of human noroviruses (NoV) in oyster GI cells. METHODS AND RESULTS: HBGA-specific monoclonal antibodies (MAbs) were used to determine the presence of the corresponding HBGA in oyster GI cells. All oyster samples tested contained type A-like HBGA in GI tissue as measured by ELISA. Recombinant Norwalk virus viral like particles (rNVLP) were bound to plates coated with oyster GI homogenate. The binding was inhibited when rNVLPs were pre-incubated with MAbs specific for type A HBGA, or samples of human saliva from type A individuals. Co-localization of rNVLP and type A-like HBGA, but not type B-like or type H-like HBGA, on GI epithelial cells was observed by immunofluorescent histochemical staining and three-channel confocal scanning laser microscopy. CONCLUSION: Type A-like HBGA is present in oyster GI cells and responsible for binding of rNVLP. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of the presence of type A-like HBGA in oyster GI cells and the specific binding of rNVLP to type A-like HBGA on oyster GI cells. The results of this study suggest that human NoV concentrate in oyster GI cells by specific binding to concentrated type A-like HBGA rather than by a nonmolecular entrapment within the tissues. PMID- 17083712 TI - Separation of yeast cells from aqueous solutions using magnetically stabilized fluidized beds. AB - AIMS: To separate Saccharomyces cerevisiae cells from aqueous solutions using magnetically stabilized fluidized beds (MSFB) that utilize a horizontal magnetic field, and to study the effect of some parameters, such as bed porosity and height, liquid flow rate and inlet concentration on cell removal efficiency and breakthrough curves. METHODS AND RESULTS: The separation process was conducted in an MSFB under the effect of horizontal magnetic field. The magnetic particles used consist of a ferromagnetic core of magnetite (Fe3O4) covered by a stable layer of activated carbon to adsorb the yeast cells from the suspension. The yeast cell concentration in the effluent was determined periodically by measuring the absorbance at 610 nm. The effect of the magnetic field intensity on the bed porosity and consequently the exit-normalized cell concentration from the bed was studied. It was found that bed porosity increased by 75%, and the normalized cell concentration in the bed effluent decreased by 30%, when the magnetic field intensity was increased from 0 to 110 mT. In addition, increasing the magnetic field intensity and bed height delayed the breakthrough point, and allowed efficient cell removal. These results demonstrate an improved method to separate cells of low concentration from cell suspension. CONCLUSIONS: This study allows the continuous separation of yeast cells from aqueous solutions in an MSFB. The removal efficiency is affected by different parameters including the bed height, flow rate and initial concentration. The removal efficiency reaches 82%, and could be improved by varying the operational parameters. SIGNIFICANCE AND IMPACT OF THE STUDY: The results obtained in this investigation show that the MSFB using horizontal fields represents a potential tool for the continuous separation of cell suspension from aqueous solution. This study will contribute to a better understanding of the hydrodynamic parameters on the separation efficiencies of the cell. PMID- 17083713 TI - The application of the herbicide bromoxynil to a model soil-derived bacterial community: impact on degradation and community structure. AB - AIMS: Bromoxynil degradation by soil micro-organisms has been shown to be co oxidative in character. In this study, we investigate both the impact of the application of increasing bromoxynil concentrations on soil-derived bacterial communities and how these changes are reflected in the degradation of the compound. Our aim was to test the hypothesis that the addition of bromoxynil to a soil-derived bacterial community, and the availability of a readily utilizable carbon source would have an impact on bromoxynil degradation, and that would be reflected in the bacteria present in the soil community. METHODS AND RESULTS: Degradation of bromoxynil was observed in soil-derived communities containing 15 mg l(-1), but not 50 mg l(-1) of the compound, unless glucose was added. This suggests that the addition of carbon stimulates co-oxidative bromoxynil degradation by the members of the bacterial community. Measurable changes in the bacterial community indicated that the addition of bromoxynil led to deterministic selection on the bacterial population, i.e. the communities observed arise through the selection of specific micro-organisms that are best adapted to the conditions in the soil. The addition of bromoxynil was also shown to have a negative impact on the presence of alpha and gamma-proteobacteria in the soil community. CONCLUSION: Bromoxynil degradation is significantly inhibited in bacterial soil communities in the absence of readily accessible carbon. The application of bromoxynil appears to exert deterministic selection on the bacterial community. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights the effects of increasing bromoxynil concentrations on a model bacterial population derived from soil. Soil communities show qualitative and quantitative differences to bromoxynil application depending on the availability of organic carbon. These findings might have implications for the persistence of bromoxynil in agricultural soils. PMID- 17083714 TI - Efficacy of electrolysed oxidizing water in inactivating Vibrio parahaemolyticus on kitchen cutting boards and food contact surfaces. AB - AIM: To determine the efficacy of electrolysed oxidizing (EO) water in inactivating Vibrio parahaemolyticus on kitchen cutting boards and food contact surfaces. METHODS AND RESULTS: Cutting boards (bamboo, wood and plastic) and food contact surfaces (stainless steel and glazed ceramic tile) were inoculated with V. parahaemolyticus. Viable cells of V. parahaemolyticus were detected on all cutting boards and food contact surfaces after 10 and 30 min, respectively, at room temperatures. Soaking inoculated food contact surfaces and cutting boards in distilled water for 1 and 3 min, respectively, resulted in various reductions of V. parahaemolyticus, but failed to remove the organism completely from surfaces. However, the treatment of EO water [pH 2.7, chlorine 40 ppm, oxidation-reduction potential 1151 mV] for 30, 45, and 60 s, completely inactivated V. parahaemolyticus on stainless steel, ceramic tile, and plastic cutting boards, respectively. CONCLUSIONS: EO water could be used as a disinfecting agent for inactivating V. parahaemolyticus on plastic and wood cutting boards and food contact surfaces. SIGNIFICANCE AND IMPACT OF THE STUDY: Rinsing the food contact surfaces with EO water or soaking cutting boards in EO water for up to 5 min could be a simple strategy to reduce cross-contamination of V. parahaemolyticus during food preparation. PMID- 17083715 TI - A novel multiplex PCR assay for the detection of Salmonella enterica serovar Enteritidis in human faeces. AB - AIMS: To develop a multiplex PCR assay for the detection of Salmonella enterica serovar Enteritidis in human faeces. METHODS AND RESULTS: A total of 54 Salmonella strains representing 19 serovars and non-Salmonella strains representing 11 different genera were used. Five primer pairs were employed in the assay. Three of them targeted to the genes hilA, spvA and invA that encode virulence-associated factors. A fourth primer pair amplified a fragment of a unique sequence within S. enterica serovar Enteritidis genomes. An internal amplification control (a fragment of a conservative sequence within the 16S rRNA genes) was targeted by a fifth primer pair. The assay produced two or three amplicons from the invA, hilA and 16S rRNA genes for 19 Salmonella serovars. All Salmonella and non-Salmonella strains yielded a band of an internal amplification control. For S. enterica serovar Typhimurium, four products (the fourth from the spvA gene), and for S. enterica serovar Enteritidis five amplicons (the fifth from the sdf gene) were observed. S. enterica serovar Enteritidis was cultured from three of 71 rectal swabs from diarrhoeal patients. Five specific amplicons were generated with the multiplex PCR assay only from culture-positive faecal samples. CONCLUSION: The multiplex PCR assay specifically detects S. enterica serovar Enteritidis. SIGNIFICANCE AND IMPACT OF THE STUDY: This is a novel multiplex PCR assay, which contains an internal amplification control and enables concurrent survey for Salmonella virulence genes. PMID- 17083717 TI - Peritoneal carcinomatosis from a small bowel carcinoid tumour. AB - BACKGROUND: Peritoneal carcinomatosis from a gastrointestinal carcinoid tumour is rare and the long-term management and prognosis have not been clearly defined. The natural history is different from gastrointestinal adenocarcinoma, although its capacity to invade regional lymph nodes and generate distal metastasis can make the management more complex. Whilst the development of carcinomatosis is uncommonly reported, it may be higher than expected. CASE PRESENTATION: A 63 years-old woman underwent emergency surgery in 1993 for right iliac fossa pain and a mass that was found to be an ileal carcinoid tumour. Over the next ten years, further surgery was required for disseminated disease with peritoneal carcinomatosis and liver metastasis. Systemic chemotherapy had little effect, although Somatostatin was used effectively to relieve symptoms caused by the disseminated disease (flushing and diarrhoea). CONCLUSION: Peritoneal carcinomatosis from carcinoid tumours is not well documented in the literature. Aggressive surgery must be performed in order to control the disease since chemotherapy has not been reported to be effective. With repeated surgery long term survival can be achieved in these patients. PMID- 17083718 TI - Target organ damage and cardiovascular complications in patients with hypertension and type 2 diabetes in Spain: a cross-sectional study. AB - BACKGROUND: Target organ damage (mainly cardiac and renal damage) is easy to evaluate in outpatient clinics and offers valuable information about patient's cardiovascular risk. The purpose of this study was to evaluate, using simple methods, the prevalence of cardiac and renal damage and its relationship to the presence of established cardiovascular disease (CVD), in patients with hypertension (HT) and type 2 diabetes mellitus (DM). METHODS: The RICARHD study is a multicentre, cross-sectional study made by 293 investigators in Nephrology and Internal Medicine Spanish outpatient clinics, and included patients aged 55 years or more with HT and type 2 DM with more than six months of diagnosis. Demographic, clinical and biochemical data, and CVD were collected from the clinical records. Cardiac damage was defined by the presence of electrocardiographic left ventricular hypertrophy (ECG-LVH), and renal damage by a calculated glomerular filtration rate (GFR) of <60 ml/min/1.73 m2, and/or the presence of an albumin/creatinine ratio > or = 30 mg/g; or an urinary albumin excretion (UAE) > or = 30 mg/24 hours. RESULTS: 2339 patients (mean age 68.9 years, 48.2% females, 51.3% with established CVD) were included. ECG-LVH was present in 22.9% of the sample, GFR <60 ml/min/1.73 m2 in 45.1%, and abnormal UAE in 58.7%. Compared with the reference patients (those without neither cardiac nor renal damage), patients with ECG-LVH alone (OR 2.20, [95%CI 1.43-3.38]), or kidney damage alone (OR 1.41, [1.13-1.75]) showed an increased prevalence of CVD. The presence of both ECG-LVH and renal damage was associated with the higher prevalence (OR 3.12, [2.33-4.19]). After stratifying by gender, this relationship was present for both, men and women. CONCLUSION: In patients with HT and type 2 DM, ECG-LVH or renal damage, evaluated using simple methods, are associated with an increased prevalence of established CVD. The simultaneous presence of both cardiac and renal damage was associated to the higher prevalence of CVD, affording complementary information. A systematic assessment of cardiac and renal damage complements the risk assessment of these patients with HT and type 2 DM. PMID- 17083719 TI - Estimating health-adjusted life expectancy conditional on risk factors: results for smoking and obesity. AB - BACKGROUND: Smoking and obesity are risk factors causing a large burden of disease. To help formulate and prioritize among smoking and obesity prevention activities, estimations of health-adjusted life expectancy (HALE) for cohorts that differ solely in their lifestyle (e.g. smoking vs. non smoking) can provide valuable information. Furthermore, in combination with estimates of life expectancy (LE), it can be tested whether prevention of obesity and smoking results in compression of morbidity. METHODS: Using a dynamic population model that calculates the incidence of chronic disease conditional on epidemiological risk factors, we estimated LE and HALE at age 20 for a cohort of smokers with a normal weight (BMI < 25), a cohort of non-smoking obese people (BMI>30) and a cohort of 'healthy living' people (i.e. non smoking with a BMI < 25). Health state valuations for the different cohorts were calculated using the estimated disease prevalence rates in combination with data from the Dutch Burden of Disease study. Health state valuations are multiplied with life years to estimate HALE. Absolute compression of morbidity is defined as a reduction in unhealthy life expectancy (LE-HALE) and relative compression as a reduction in the proportion of life lived in good health (LE-HALE)/LE. RESULTS: Estimates of HALE are highest for a 'healthy living' cohort (54.8 years for men and 55.4 years for women at age 20). Differences in HALE compared to 'healthy living' men at age 20 are 7.8 and 4.6 for respectively smoking and obese men. Differences in HALE compared to 'healthy living' women at age 20 are 6.0 and 4.5 for respectively smoking and obese women. Unhealthy life expectancy is about equal for all cohorts, meaning that successful prevention would not result in absolute compression of morbidity. Sensitivity analyses demonstrate that although estimates of LE and HALE are sensitive to changes in disease epidemiology, differences in LE and HALE between the different cohorts are fairly robust. In most cases, elimination of smoking or obesity does not result in absolute compression of morbidity but slightly increases the part of life lived in good health. CONCLUSION: Differences in HALE between smoking, obese and 'healthy living' cohorts are substantial and similar to differences in LE. However, our results do not indicate that substantial compression of morbidity is to be expected as a result of successful smoking or obesity prevention. PMID- 17083720 TI - The co-distribution of Plasmodium falciparum and hookworm among African schoolchildren. AB - BACKGROUND: Surprisingly little is known about the geographical overlap between malaria and other tropical diseases, including helminth infections. This is despite the potential public health importance of co-infection and synergistic opportunities for control. METHODS: Statistical models are presented that predict the large-scale distribution of hookworm in sub-Saharan Africa (SSA), based on the relationship between prevalence of infection among schoolchildren and remotely sensed environmental variables. Using a climate-based spatial model of the transmission potential for Plasmodium falciparum malaria, adjusted for urbanization, the spatial congruence of populations at coincident risk of infection is determined. RESULTS: The model of hookworm indicates that the infection is widespread throughout Africa and that, of the 179.3 million school aged children who live on the continent, 50.0 (95% CI: 48.9-51.1) million (27.9% of total population) are infected with hookworm and 45.1 (95% CI: 43.9-46) million are estimated to be at risk of coincident infection. CONCLUSION: Malaria and hookworm infection are widespread throughout SSA and over a quarter of school aged children in sub-Saharan Africa appear to be at risk of coincident infection and thus at enhanced risk of clinical disease. The results suggest that the control of parasitic helminths and of malaria in school children could be viewed as essential co-contributors to promoting the health of schoolchildren. PMID- 17083721 TI - The cell biology of HIV-1 and other retroviruses. AB - In recognition of the growing influence of cell biology in retrovirus research, we recently organized a Summer conference sponsored by the American Society for Cell Biology (ASCB) on the Cell Biology of HIV-1 and other Retroviruses (July 20 23, 2006, Emory University, Atlanta, Georgia). The meeting brought together a number of leading investigators interested in the interplay between cell biology and retrovirology with an emphasis on presentation of new and unpublished data. The conference was arranged from early to late events in the virus replication cycle, with sessions on viral fusion, entry, and transmission; post-entry restrictions to retroviral infection; nuclear import and integration; gene expression/regulation of retroviral Gag and genomic RNA; and assembly/release. In this review, we will attempt to touch briefly on some of the highlights of the conference, and will emphasize themes and trends that emerged at the meeting. MEETING REPORT: The conference began with a keynote address from W. Sundquist on the biochemistry of HIV-1 budding. This presentation will be described in the section on Assembly and Release of Retroviruses. PMID- 17083722 TI - GP-initiated preconception counselling in a randomised controlled trial does not induce anxiety. AB - BACKGROUND: Preconception counselling (PCC) can reduce adverse pregnancy outcome by addressing risk factors prior to pregnancy. This study explores whether anxiety is induced in women either by the offer of PCC or by participation with GP-initiated PCC. METHODS: Randomised trial of usual care versus GP-initiated PCC for women aged 18-40, in 54 GP practices in the Netherlands. Women completed the six-item Spielberger State Trait Anxiety Inventory (STAI) before PCC (STAI-1) and after (STAI-2). After pregnancy women completed a STAI focusing on the first trimester of pregnancy (STAI-3). RESULTS: The mean STAI-1-score (n = 466) was 36.4 (95% CI 35.4-37.3). Following PCC there was an average decrease of 3.6 points in anxiety-levels (95% CI, 2.4-4.8). Mean scores of the STAI-3 were 38.5 (95% CI 37.7-39.3) in the control group (n = 1090) and 38.7 (95% CI 37.9-39.5) in the intervention group (n = 1186). CONCLUSION: PCC from one's own GP reduced anxiety after participation, without leading to an increase in anxiety among the intervention group during pregnancy. We therefore conclude that GPs can offer PCC to the general population without fear of causing anxiety. PMID- 17083723 TI - Stromal derived factor-1 (SDF-1/CXCL12) and CXCR4 in renal cell carcinoma metastasis. AB - Renal cell carcinoma (RCC) is characterized by organ-specific metastases. The chemokine stromal derived factor-1 (SDF-1/CXCL12) and its receptor CXCR4 have been suggested to regulate organ-specific metastasis in various other cancers. On this basis, we hypothesized that the biological axis of CXCL12 via interaction with its receptor, CXCR4, is a major mechanism for RCC metastasis. We demonstrated that CXCR4 was significantly expressed on circulating cytokeratin+ RCC cells from patients with known metastatic RCC. We detected up-regulation of CXCR4 mRNA and protein levels on a human RCC cell line by either knockdown of the von Hippel-Lindau (VHL) tumor suppressor protein, or incubating the cells under hypoxic conditions. The enhanced CXCR4 expression was mediated through the interaction of the Hypoxia Inducible Factor-1alpha (HIF-1alpha) with the promoter region of the CXCR4 gene. Furthermore, the expression of CXCR4 on human RCC directly correlated with their metastatic ability in vivo in both heterotopic and orthotopic SCID mouse models of human RCC. Neutralization of CXCL12 in SCID mice abrogated metastasis of RCC to target organs expressing high levels of CXCL12; without altering tumor cell proliferation, apoptosis, or tumor-associated angiogenesis. Therefore, our data suggest that the CXCL12/CXCR4 biological axis plays an important role in regulating the organ-specific metastasis of RCC. PMID- 17083724 TI - Phosphorylation of HIV-1 Tat by CDK2 in HIV-1 transcription. AB - BACKGROUND: Transcription of HIV-1 genes is activated by HIV-1 Tat protein, which induces phosphorylation of RNA polymerase II (RNAPII) C-terminal domain (CTD) by CDK9/cyclin T1. Earlier we showed that CDK2/cyclin E phosphorylates HIV-1 Tat in vitro. We also showed that CDK2 induces HIV-1 transcription in vitro and that inhibition of CDK2 expression by RNA interference inhibits HIV-1 transcription and viral replication in cultured cells. In the present study, we analyzed whether Tat is phosphorylated in cultured cells by CDK2 and whether Tat phosphorylation has a regulatory effect on HIV-1 transcription. RESULTS: We analyzed HIV-1 Tat phosphorylation by CDK2 in vitro and identified Ser16 and Ser46 residues of Tat as potential phosphorylation sites. Tat was phosphorylated in HeLa cells infected with Tat-expressing adenovirus and metabolically labeled with 32P. CDK2-specific siRNA reduced the amount and the activity of cellular CDK2 and significantly decreased phosphorylation of Tat. Tat co-migrated with CDK2 on glycerol gradient and co-immunoprecipitated with CDK2 from the cellular extracts. Tat was phosphorylated on serine residues in vivo, and mutations of Ser16 and Ser46 residues of Tat reduced Tat phosphorylation in vivo. Mutation of Ser16 and Ser46 residues of Tat reduced HIV-1 transcription in transiently transfected cells. The mutations of Tat also inhibited HIV-1 viral replication and Tat phosphorylation in the context of the integrated HIV-1 provirus. Analysis of physiological importance of the S16QP(K/R)19 and S46YGR49 sequences of Tat showed that Ser16 and Ser46 and R49 residues are highly conserved whereas mutation of the (K/R)19 residue correlated with non-progression of HIV-1 disease. CONCLUSION: Our results indicate for the first time that Tat is phosphorylated in vivo; Tat phosphorylation is likely to be mediated by CDK2; and phosphorylation of Tat is important for HIV-1 transcription. PMID- 17083725 TI - Effect of Gabapentin on morphine demand and pain after laparoscopic sterilization using Filshie clips. A double blind randomized clinical trial. AB - BACKGROUND: A considerable number of patients require opioids during recovery after laparoscopic sterilization. This implies nausea, dizziness and sedation and increases the number of unplanned admissions. Gabapentin has shown excellent postoperative analgesic effect in a number of recent studies with few side effects. This study was designed to test whether gabapentin given preoperatively can reduce the number of patients needing morphine in the recovery period. METHODS: 80 females scheduled for laparoscopic sterilization using Filshie clips were randomized to two treatment groups (Gaba group and control group). All patients received lornoxicam 8 mg p.o. 30 min. before the procedure. Patients in the Gaba group received gabapentin 1200 mg p.o. and patients in the control group received placebo capsules prior to the procedure. All patients were anesthetized according to a protocol, using remifentanil and propofol. Postoperative analgesia was obtained with patient controlled infusion of morphine. Pain, nausea, dizziness and sedation were scored at 2 and 4 hours after end of anesthesia. The expenditure of morphine was the primary measure for the effect of analgesia and the number of patients demanding morphine was the primary endpoint. RESULTS: Three patients were excluded because of procedural errors and one because of conversion to open surgery. 38 patients completed the study in each group.32 (84%) patients in the gabapentin group and 37 (97%) patients in the control group did require morphine in the recovery period. (p = 0,049). There was no significant difference between mean morphine consumption, pain scores and frequency of adverse effects (nausea, dizziness, sedation and vomiting) CONCLUSION: The postoperative analgesic effect of gabapentin given preoperatively was confirmed in this study. For this procedure, with pain predominantly in the immediate recovery period, and of less intensity than after major surgical procedures, the effect demonstrated is much less pronounced than in similar studies of major surgery. General use of gabapentin as analgesic for laparoscopic sterilization is not supported by this study. TRIAL REGISTRATION: Current Controlled Trials ISCRTN39209275. PMID- 17083726 TI - IL-17 mRNA in sputum of asthmatic patients: linking T cell driven inflammation and granulocytic influx? AB - BACKGROUND: The role of Th2 cells (producing interleukin (IL-)4, IL-5 and IL-13) in allergic asthma is well-defined. A distinct proinflammatory T cell lineage has recently been identified, called Th17 cells, producing IL-17A, a cytokine that induces CXCL8 (IL-8) and recruits neutrophils. Neutrophilic infiltration in the airways is prominent in severe asthma exacerbations and may contribute to airway gland hypersecretion, bronchial hyper-reactivity and airway wall remodelling in asthma. AIM: to study the production of IL-17 in asthmatic airways at the mRNA level, and to correlate this with IL-8 mRNA, neutrophilic inflammation and asthma severity. METHODS: We obtained airway cells by sputum induction from healthy individuals (n = 15) and from asthmatic patients (n = 39). Neutrophils were counted on cytospins and IL-17A and IL-8 mRNA expression was quantified by real time RT-PCR (n = 11 controls and 33 asthmatics). RESULTS: Sputum IL-17A and IL-8 mRNA levels are significantly elevated in asthma patients compared to healthy controls. IL-17 mRNA levels are significantly correlated with CD3gamma mRNA levels in asthmatic patients and mRNA levels of IL-17A and IL-8 correlated with each other and with sputum neutrophil counts. High sputum IL-8 and IL-17A mRNA levels were also found in moderate-to-severe (persistent) asthmatics on inhaled steroid treatment. CONCLUSION: The data suggest that Th17 cell infiltration in asthmatic airways links T cell activity with neutrophilic inflammation in asthma. PMID- 17083727 TI - The cumate gene-switch: a system for regulated expression in mammalian cells. AB - BACKGROUND: A number of expression systems have been developed where transgene expression can be regulated. They all have specific characteristics making them more suitable for certain applications than for others. Since some applications require the regulation of several genes, there is a need for a variety of independent yet compatible systems. RESULTS: We have used the regulatory mechanisms of bacterial operons (cmt and cym) to regulate gene expression in mammalian cells using three different strategies. In the repressor configuration, regulation is mediated by the binding of the repressor (CymR) to the operator site (CuO), placed downstream of a strong constitutive promoter. Addition of cumate, a small molecule, relieves the repression. In the transactivator configuration, a chimaeric transactivator (cTA) protein, formed by the fusion of CymR with the activation domain of VP16, is able to activate transcription when bound to multiple copies of CuO, placed upstream of the CMV minimal promoter. Cumate addition abrogates DNA binding and therefore transactivation by cTA. Finally, an adenoviral library of cTA mutants was screened to identify a reverse cumate activator (rcTA), which activates transcription in the presence rather than the absence of cumate. CONCLUSION: We report the generation of a new versatile inducible expression system. PMID- 17083728 TI - Prevalence and trends of selected urologic conditions for VA healthcare users. AB - BACKGROUND: Conducted as part of the Urologic Diseases in America project whose aim was to quantify the burden of urologic diseases on the American public, this study focuses on Veterans Health Administration (VHA) users as a special population to supplement data on overall prevalence rates and trends in the United States. Veterans comprise 25% of the male population 18 years or older and contribute substantially to the overall burden of urologic conditions. The objective of this study is to describe the prevalence rates and trends of urologic cancers and selected benign conditions from 1999 to 2002 for VHA users. METHODS: VHA administrative files for 1999-2002 and Medicare claims files for the same years were used to identify those who had a diagnosis of qualifying urologic conditions. RESULTS: Among the conditions evaluated, prostate cancer was listed as a primary diagnosis for 5.4% of VHA users in 2002, followed in decreasing prevalence by erectile dysfunction (2.9%), renal mass (1.5%), interstitial cystitis (1.4%), and prostatitis (1.1%). Age-adjusted rates showed significant increases for renal mass (31%), interstitial cystitis (14%), and erectile dysfunction (8%) between 1999 and 2002. Systematic variations in prevalence rates and trends were observed by age, race/ethnicity, and region. Those in the Western region generally had lower age-adjusted prevalence rates and their increases were also slower than other regions. Addition of Medicare data resulted in large increases (21 to 489%) in prevalence among VHA users, suggesting substantial amount of non-VA urological care provided to VHA users. CONCLUSION: Prevalence rates for many urologic diseases increased between 1999 and 2002, which were not entirely attributable to the aging of veterans. This changing urologic disease burden has substantial implications for access to urologic care and treatment capacity, especially in light of the level of urologic care delivered to veterans by Medicare providers outside the VA. Further study on the factors associated with these increases and how they affect the patterns, cost, and quality of care in veterans is needed. PMID- 17083729 TI - Technical development of PubMed interact: an improved interface for MEDLINE/PubMed searches. AB - BACKGROUND: The project aims to create an alternative search interface for MEDLINE/PubMed that may provide assistance to the novice user and added convenience to the advanced user. An earlier version of the project was the 'Slider Interface for MEDLINE/PubMed searches' (SLIM) which provided JavaScript slider bars to control search parameters. In this new version, recent developments in Web-based technologies were implemented. These changes may prove to be even more valuable in enhancing user interactivity through client-side manipulation and management of results. RESULTS: PubMed Interact is a Web-based MEDLINE/PubMed search application built with HTML, JavaScript and PHP. It is implemented on a Windows Server 2003 with Apache 2.0.52, PHP 4.4.1 and MySQL 4.1.18. PHP scripts provide the backend engine that connects with E-Utilities and parses XML files. JavaScript manages client-side functionalities and converts Web pages into interactive platforms using dynamic HTML (DHTML), Document Object Model (DOM) tree manipulation and Ajax methods. With PubMed Interact, users can limit searches with JavaScript slider bars, preview result counts, delete citations from the list, display and add related articles and create relevance lists. Many interactive features occur at client-side, which allow instant feedback without reloading or refreshing the page resulting in a more efficient user experience. CONCLUSION: PubMed Interact is a highly interactive Web-based search application for MEDLINE/PubMed that explores recent trends in Web technologies like DOM tree manipulation and Ajax. It may become a valuable technical development for online medical search applications. PMID- 17083730 TI - Effects of brain polarization on reaction times and pinch force in chronic stroke. AB - BACKGROUND: Previous studies showed that anodal transcranial DC stimulation (tDCS) applied to the primary motor cortex of the affected hemisphere (M1affected hemisphere) after subcortical stroke transiently improves performance of complex tasks that mimic activities of daily living (ADL). It is not known if relatively simpler motor tasks are similarly affected. Here we tested the effects of tDCS on pinch force (PF) and simple reaction time (RT) tasks in patients with chronic stroke in a double-blind cross-over Sham-controlled experimental design. RESULTS: Anodal tDCS shortened reaction times and improved pinch force in the paretic hand relative to Sham stimulation, an effect present in patients with higher impairment. CONCLUSION: tDCS of M1affected hemisphere can modulate performance of motor tasks simpler than those previously studied, a finding that could potentially benefit patients with relatively higher impairment levels. PMID- 17083731 TI - Machine learning techniques in disease forecasting: a case study on rice blast prediction. AB - BACKGROUND: Diverse modeling approaches viz. neural networks and multiple regression have been followed to date for disease prediction in plant populations. However, due to their inability to predict value of unknown data points and longer training times, there is need for exploiting new prediction softwares for better understanding of plant-pathogen-environment relationships. Further, there is no online tool available which can help the plant researchers or farmers in timely application of control measures. This paper introduces a new prediction approach based on support vector machines for developing weather-based prediction models of plant diseases. RESULTS: Six significant weather variables were selected as predictor variables. Two series of models (cross-location and cross-year) were developed and validated using a five-fold cross validation procedure. For cross-year models, the conventional multiple regression (REG) approach achieved an average correlation coefficient (r) of 0.50, which increased to 0.60 and percent mean absolute error (%MAE) decreased from 65.42 to 52.24 when back-propagation neural network (BPNN) was used. With generalized regression neural network (GRNN), the r increased to 0.70 and %MAE also improved to 46.30, which further increased to r = 0.77 and %MAE = 36.66 when support vector machine (SVM) based method was used. Similarly, cross-location validation achieved r = 0.48, 0.56 and 0.66 using REG, BPNN and GRNN respectively, with their corresponding %MAE as 77.54, 66.11 and 58.26. The SVM-based method outperformed all the three approaches by further increasing r to 0.74 with improvement in %MAE to 44.12. Overall, this SVM-based prediction approach will open new vistas in the area of forecasting plant diseases of various crops. CONCLUSION: Our case study demonstrated that SVM is better than existing machine learning techniques and conventional REG approaches in forecasting plant diseases. In this direction, we have also developed a SVM-based web server for rice blast prediction, a first of its kind worldwide, which can help the plant science community and farmers in their decision making process. The server is freely available at http://www.imtech.res.in/raghava/rbpred/. PMID- 17083732 TI - In vivo activity of terpinen-4-ol, the main bioactive component of Melaleuca alternifolia Cheel (tea tree) oil against azole-susceptible and -resistant human pathogenic Candida species. AB - BACKGROUND: Recent investigations on the antifungal properties of essential oil of Melaleuca alternifolia Cheel (Tea Tree Oil, TTO) have been performed with reference to the treatment of vaginal candidiasis. However, there is a lack of in vivo data supporting in vitro results, especially regarding the antifungal properties of TTO constituents. Thus, the aim of our study was to investigate the in vitro and the in vivo anti-Candida activity of two critical bioactive constituents of TTO, terpinen-4-ol and 1,8-cineole. METHODS: Oophorectomized, pseudoestrus rats under estrogen treatment were used for experimental vaginal infection with azole (fluconazole, itraconazole) -susceptible or -resistant strains of C. albicans. All these strains were preliminarily tested for in vitro susceptibility to TTO, terpinen-4-ol and 1,8-cineole for their antifungal properties, using a modification of the CLSI (formerly NCCLS) reference M27-A2 broth micro-dilution method. RESULTS: In vitro minimal inhibitory concentrations (MIC90) values were 0.06% (volume/volume) for terpinen-4-ol and 4% (volume/volume) for 1,8-cineole, regardless of susceptibility or resistance of the strains to fluconazole and itraconazole. Fungicidal concentrations of terpinen-4-ol were equivalent to the candidastatic activity. In the rat vaginal infection model, terpinen-4-ol was as active as TTO in accelerating clearance from the vagina of all Candida strains examined. CONCLUSION: Our data suggest that terpinen-4-ol is a likely mediator of the in vitro and in vivo activity of TTO. This is the first in vivo demonstration that terpinen-4-ol could control C. albicans vaginal infections. The purified compound holds promise for the treatment of vaginal candidiasis, and particularly the azole-resistant forms. PMID- 17083733 TI - Complex host-pathogen coevolution in the Apterostigma fungus-growing ant-microbe symbiosis. AB - BACKGROUND: The fungus-growing ant-microbe symbiosis consists of coevolving microbial mutualists and pathogens. The diverse fungal lineages that these ants cultivate are attacked by parasitic microfungi of the genus Escovopsis. Previous molecular analyses have demonstrated strong phylogenetic congruence between the ants, the ants-cultivated fungi and the garden pathogen Escovopsis at ancient phylogenetic levels, suggesting coevolution of these symbionts. However, few studies have explored cophylogenetic patterns between these symbionts at the recent phylogenetic levels necessary to address whether these parasites are occasionally switching to novel hosts or whether they are diversifying with their hosts as a consequence of long-term host fidelity. RESULTS: Here, a more extensive phylogenetic analysis of Escovopsis lineages infecting the gardens of Apterostigma ants demonstrates that these pathogens display patterns of phylogenetic congruence with their fungal hosts. Particular clades of Escovopsis track particular clades of cultivated fungi, and closely-related Escovopsis generally infect closely-related hosts. Discordance between host and parasite phylogenies, however, provides the first evidence for occasional host-switches or acquisitions of novel infections from the environment. CONCLUSION: The fungus growing ant-microbe association has a complex coevolutionary history. Though there is clear evidence of host-specificity on the part of diverse Escovopsis lineages, these pathogens have switched occasionally to novel host fungi. Such switching is likely to have profound effects on how these host and parasites adapt to one another over evolutionary time scales and may impact how disease spreads over ecological time scales. PMID- 17083734 TI - A new shape for an old function: lasting effect of a physiologic surgical restoration of the left ventricle. AB - BACKGROUND: Long-term morphofunctional outcome may vary widely in surgical anterior left ventricular wall restoration, suggesting variability in post surgical remodeling similar to that observed following acute myocardial infarction. The aim of this pilot study was to demonstrate that surgical restoration obtained with a particular shape of endoventricular patch leads to steady morphofunctional ventricular improvement when geometry, volume and residual akinesia can be restored as normal as possible. METHODS: This study involved 12 consecutive patients with previous anterior myocardial infarction, dilated cardiomyopathy and no mitral procedures, who underwent left ventricular reconstruction and coronary revascularization between May 2002 and May 2003 using a small, narrow, oval patch aiming at a volume 18 years) male slum dwellers (mean age=34.6+/-14.4 years) of Midnapore town, West Bengal, India, was undertaken to study the inter-relationships of chronic energy deficiency (CED), monthly family income (MFI), self-reported morbidity and hospitalization due to severe illness. The mean height, weight and body mass index (BMI) of the subjects were 160.0 cm, 50.8 kg and 19.9 kg/m2, respectively. The overall frequencies of CED (BMI<18.5 kg/m2), morbidity and hospitalization were 38.2%, 34.4% and 13.7%, respectively. Based on the WHO classification, the prevalence of CED among this population was high (20-39%), indicating a serious situation. Overall, MFI was significantly (p<0.01) positively correlated with BMI (r=0.21). Linear regression analyses showed that MFI had a significant impact (t=3.08; p<0.002) on BMI. Overall, MFI explained 3.9% variation in BMI. Subjects belonging to the lowest family income group (FIG I) had the lowest mean BMI (19.1 kg/m2) and the highest rate of CED (46.3%) and morbidity (36.6%). Those in the highest family income group (FIG III) had the largest mean BMI (20.8 kg/m2) and lowest rate of CED (30.2%) and morbidity (30.2%). The highest rate (18.9%) of hospitalization was found in this group. There were significant family income group differences in mean BMI (F=3.134, p<0.05). The frequency of morbidity (24.6%) and hospitalization (11.9%) was lowest among normal BMI individuals. Morbidity was significantly higher (chi2=11.92, p=0.0026) among CED (48.2%) subjects compared with normal BMI individuals (OR=2.85; CI=1.49-5.46). Similarly, compared with normal BMI subjects, morbidity was higher (38.5%; OR=1.92; 95% CI=0.50-7.18) among overweight subjects. Hospitalization was more common among CED subjects (16.1%; OR=1.42; CI=0.58-3.45) compared with normal BMI subjects. Similarly, the frequency of hospitalization was more among overweight individuals (15.4%; OR=1.35; 95% CI=0.0-7.59). In conclusion, this study provides evidence that the frequency of CED among this population is high, indicating a serious situation. Moreover there exists strong inter-relationships between BMI, CED, MFI and morbidity. PMID- 17083745 TI - The role of myristoylation in the membrane association of the Lassa virus matrix protein Z. AB - The Z protein is the matrix protein of arenaviruses and has been identified as the main driving force for budding. Both LCMV and Lassa virus Z proteins bud from cells in the absence of other viral proteins as enveloped virus-like particles. Z accumulates near the inner surface of the plasma membrane where budding takes place. Furthermore, biochemical data have shown that Z is strongly membrane associated. The primary sequence of Z lacks a typical transmembrane domain and until now it is not understood by which mechanism Z is able to interact with cellular membranes. In this report, we analyzed the role of N-terminal myristoylation for the membrane binding of Lassa virus Z. We show that disruption of the N-terminal myristoylation signal by substituting the N-terminal glycine with alanine (Z-G2A mutant) resulted in a significant reduction of Z protein association with cellular membranes. Furthermore, removal of the myristoylation site resulted in a relocalization of Z from a punctuate distribution to a more diffuse cellular distribution pattern. Finally, treatment of Lassa virus-infected cells with various myristoylation inhibitors drastically reduced efficient Lassa virus replication. Our data indicate that myristoylation of Z is critical for its binding ability to lipid membranes and thus, for effective virus budding. PMID- 17083746 TI - Factors associated with intentions to adhere to colorectal cancer screening follow-up exams. AB - BACKGROUND: To increase adherence rate to recommendations for follow-up after abnormal colorectal cancer (CRC) screening results, factors that inhibit and facilitate follow-up must be identified. The purpose of this study was to identify the factors associated with intention to adhere to CRC screening follow up exams. METHODS: During a 4-week period in October 2003, this survey was conducted with 426 subjects participating in a community-based CRC screening program in Nagano, Japan. Study measures included intention to adhere to recommendation for clinical follow-up in the event of an abnormal fecal occult blood test (FOBT) result, perceived susceptibility and severity of CRC, perceived benefits and barriers related to undergoing follow-up examination, social support, knowledge of CRC risk factors, health status, previous CRC screening, personality and social demographic characteristics. Univariate and multivariate logistic regression analyses on intention to adhere to recommendations for follow up were performed. RESULTS: Among the 288 individuals analyzed, approximately 74.7% indicated that they would definitely adhere to recommendations for follow up. After controlling for age, gender, marital status, education, economic status, trait anxiety, bowel symptoms, family history of CRC, and previous screening FOBT, analyses revealed that lower levels of perceived barriers, higher levers of perceived benefits and knowledge of CRC risk factors were significantly associated with high intention respectively. CONCLUSION: The results of this study suggest that future interventions should focus on reducing modifiable barriers by clarifying misperceptions about follow-up, promoting the acceptance of complete diagnostic evaluations, addressing psychological distress, and making follow-up testing more convenient and accessible. Moreover, educating the public regarding the risk factors of CRC and increasing understanding of the benefits of follow-up is also important. PMID- 17083748 TI - The epidemiology of atypical mycobacterial diseases in northern England: a space time clustering and Generalized Linear Modelling approach. AB - The incidence of infection by mycobacteria, other than tubercle bacilli (MOTT) is increasing in the United Kingdom, Europe and the United States. These diseases increase morbidity and are an increasing public health concern. However, the epidemiology of disease due to these species is not well characterized. We used space-time clustering approaches and Generalized Linear Modelling to investigate the potential predictors of disease in cases of infection by organisms of the Mycobacterium avium complex (MAC) and M. malmoense recorded in the north of England during 2000-2005. There was significant spatial and temporal clustering in juvenile cases of infection by MAC but not for cases of infection in adults by either species. There were no significant predictors of infection by M. malmoense or juvenile cases of M. avium. Incidence of disease caused by M. avium in adults was significantly related to health deprivation and weakly related to rainfall. We consider possible reasons for the difference in epidemiology in infection by M. avium in adults and juveniles. PMID- 17083749 TI - Correlates of length of stay, cost of care, and mortality among patients hospitalized for necrotizing fasciitis. AB - Several previous studies of necrotizing fasciitis (NF) have been single institution investigations suffering from small samples sizes. This study of 216 NF patients hospitalized in Florida, USA, during 2001 was designed to identify risk factors for length of stay (LOS), total patient charges (TC), and mortality, using a statewide database. Robust gamma mixed regression was used to determine the predictors of LOS and TC while simultaneously accounting for outliers and the clustering of patients in 87 hospitals. Relative risks (RR) for hospital mortality were calculated using binomial regression. The NF hospitalization rate in Florida was 1.3/100,000. The median TC was US$54,533 and cumulative charges for all 216 patients were nearly US$20 million. Patients aged > or =44 years at the time of admission were five times as likely to expire in the hospital than patients who were aged < or =43 years (adjusted RR 5.08, P=0.03). Unexpectedly, diabetes was associated with a 61% reduction in the risk of hospital mortality (adjusted RR 0.39, P=0.04). Age > or =44 years was the most powerful predictor of prolonged LOS, elevated TC, and an increased risk of hospital mortality in patients suffering from NF. PMID- 17083750 TI - Survival rate of self-tapping implants for bone-anchored hearing aids. AB - OBJECTIVE: The purpose of this study was to investigate whether a new self tapping implant for a bone-anchored hearing aid (BAHA) had the same high frequency of osseointegration as previous implants requiring pre-tapping. METHOD: Over a three-year period, 144 consecutive implants were placed in the mastoid for BAHA and evaluated. RESULTS: Two implants were lost; both were of the self tapping type. One was in an 11-year-old boy, who lost his implant six weeks after surgery when the BAHA was fitted. The other was in an elderly man, a heavy smoker with diabetes. Using Fisher's exact test, there was no significant difference between the two groups (p>0.30). CONCLUSION: Self-tapping implants facilitate surgery and shorten operating time. Over a short follow up, we did not find any significant difference; however, it is important to follow these implants over a longer time period. PMID- 17083751 TI - Recurrent upper airway infections and bacterial biofilms. AB - BACKGROUND: Bacterial biofilms identified in various medical devices used in otorhinolaryngology, including tympanostomy tubes, voice prostheses, and cochlear implants, can directly colonise mucosal tissues. The upper airways seem to be at high risk for this type of colonisation. Chronic and/or recurrent upper airway infections may be related to the complex structural and biochemical (quorum sensing) organisation of the biofilm which interferes with the activity of antibiotics (including those with proven in vitro efficacy), thus promoting the establishment of a chronic infection eradicable only by surgical treatment. Biofilm formation plays a role in upper respiratory infections: it not only explains the resistance of these infections to antibiotic therapy but it also represents an important element that contributes to the maintenance of a chronic inflammatory reaction. OBJECTIVES: To document the presence of biofilms in surgical tissue specimens from patients with recurrent infection diseases, and identify their possible role in the chronicity of these infectious processes. METHOD: We examined 32 surgical specimens from the upper respiratory tract (tonsils, adenoids, mucosa from the ethmoid and maxillary sinuses) of 28 patients (20 adults, eight children) with upper airway infections that had persisted despite repeated treatment with anti-inflammatory agents and antibiotics with demonstrated in vitro efficacy. Tissues were cultured using conventional methods and subjected to scanning electron microscopy for detection of biofilm formation. RESULTS: Over 80 per cent (26/32; 81.3 per cent) of the tissue specimens were culture-positive. Bacterial biofilms (associated in most cases with coccoid bacteria) were observed in 65.6 per cent of the tissue samples. PMID- 17083752 TI - A prospective comparative study to examine the effects of oral diazepam on blood pressure and anxiety levels in patients with acute epistaxis. AB - OBJECTIVE: To examine the effects of oral diazepam on blood pressure and anxiety in patients with acute epistaxis. STUDY DESIGN AND SETTING: A prospective comparative study in an otorhinolaryngology tertiary referral centre. PARTICIPANTS: Patients with acute epistaxis requiring hospital admission. INTERVENTION: Oral diazepam. MAIN OUTCOME MEASURES: Anxiety and blood pressure levels. RESULTS: 32 patients received diazepam and 45 did not (control). On average, patients were hypertensive on admission (mean [standard deviation (SD)] systolic blood pressure diazepam group=157 mmHg [26], control=152 mmHg [23]; diastolic blood pressure diazepam group=87 mmHg [16], control=87 mmHg [18]). Both groups showed significant blood pressure reduction on discharge (p<0.0001) but the difference in mean blood pressure reduction between the two groups was insignificant (systolic blood pressure p=0.16, 95% confidence interval [CI]=-5 to +19 mmHg; diastolic blood pressure p=0.43, 95% CI=-8 to +10 mmHg). Anxiety was significantly lower on discharge (p<0.0001) but the difference in mean fall in anxiety scores between the two groups was insignificant (p=0.08, 95% CI=0 to +2). There was no significant correlation between total diazepam and changes in blood pressure (systolic blood pressure p=0.32; diastolic blood pressure p=0.65) or anxiety (p=0.73), nor between blood pressure and anxiety on admission (systolic blood pressure p=0.45; diastolic blood pressure p=0.72). CONCLUSIONS: Elevated blood pressure and anxiety in acute epistaxis patients reduced on epistaxis resolution irrespective of oral diazepam use. The elevated blood pressure does not appear to be directly related to anxiety. PMID- 17083753 TI - What are the illness perceptions of people with dysphonia: a pilot study. AB - BACKGROUND: Patients do not respond to treatment in a predictable manner. Individual preconceptions determine help seeking, compliance and treatment outcome, yet clinicians rarely explore these issues. The illness perception approach sees the patient as an active participant in the healthcare process. AIMS: The aim of this study was to investigate the illness perceptions of people with dysphonia. The subsidiary aims were to correlate the Illness Perception Questionnaire with any psychological distress identified and a self-report measure of dysphonia, and to consider any potential implications for patient management. DESIGN: Prospective, cross-sectional observation. SETTING: Primary and secondary care, two general and four community hospitals. PARTICIPANTS: Fifty adult patients with dysphonia due to benign disease completed three self administered questionnaires, which investigated their illness perceptions, psychological distress and perceptions of the impact of the presenting 'illness'. MEASURES: The dysphonia was categorised as being due to functional (n=40) or organic (n=10) causes. All the voices were rated by an expert listener according to the GRBAS (grade, roughness, breathiness, aesthenia, strain) scale. PARTICIPANTS completed the Illness Perception Questionnaire, the Vocal Performance Questionnaire and the Hospital Anxiety and Depression scale. RESULTS: Patients showed a wide variation in perception of causation. They had no strong perceptions about the causes, consequences or duration of the presenting dysphonia. Functional dysphonics reported greater consequences, lower perceived control and increased anxiety when compared to patients with organic dysphonia. In terms of cure/control, all patients expected treatment to be helpful but this expectancy reduced as time increased. Anxiety was more associated with functional dysphonia, however, only 17 per cent of the subjects in this group showed clinically significant levels of signs of psychological distress. CONCLUSIONS: Lay illness representations often diverge from the clinician's understanding of the presenting problem and strongly influence treatment behaviour. Early exploration of illness perceptions may enhance health behaviour and maximise the impact of intervention. PMID- 17083754 TI - Individual differences of acupuncture analgesia in humans using cDNA microarray. AB - A large amount of evidence suggests that acupuncture stimulation enhances the experimental pain threshold in various animal models. Acupuncture analgesia is mediated by the endogenous opioid system, and the analgesic response to acupuncture shows individual variation. This study identified and characterized the genes that differ between high responders (HR) and low responders (LR) on acupuncture stimulation, using a cDNA microarray. Fifteen participants were stimulated at the LI 4 acupuncture point, and the finger withdrawal latency (FWL) test was performed to classify the HR and LR groups. Total RNA was then extracted from blood samples from each group and used as a template to synthesize cDNA. The cDNA was applied to Code Link UniSet Human 20K microarray chips. The Symptom Checklist 90-Revised (SCL-90-R) was also analyzed as a measure of psychological variation. The FWL was significantly elevated in the HR group after acupuncture stimulation, whereas there was little increase in the LR group. The ratio of HR to LR subjects was 9:6. We found that 353 and 22 genes were up- and downregulated, respectively, in the HR group. However, the SCL-90-R profiles did not differ significantly between the two groups. These results suggest that the individual variation in acupuncture analgesia, verified by measuring the FWL in the HR and LR groups, resulted from genetic inheritance rather than differences in the psychological environment. PMID- 17083755 TI - Complexities in defining remission in rheumatic diseases. AB - The rheumatology community has devoted increasing attention to the subject of remission over the past 2 decades, on the basis of greater appreciation of the long-term severity of inflammatory rheumatic diseases and availability of new therapies and approaches to improve outcomes. Nonetheless, description of remission in rheumatic diseases is complex, compared to many nonrheumatic diseases. Recognition of remission requires a set of measures or an index rather than a single "gold standard." Spontaneous remission is not infrequent in people with early inflammatory arthritis, including some who may meet criteria for rheumatoid arthritis (RA) over less than a few months, and may be confused with a drug-induced remission. Remission may be transient in many patients over short periods, and the length of time required to maintain remission status varies in different reports. Maintenance of a state of remission in autoimmune diseases that result from dysregulatory processes, rather than invasion of foreign cells or toxins, generally requires ongoing therapy indefinitely. Patients who have organ damage or functional disability may be described as "in remission," although they are free of disease activity only, but not necessarily free of disease consequences. A status of "low disease activity" or "near remission" with 70% to 90% of the features of an ideal remission may be adequate for many people with rheumatic diseases to avoid risks that may be required to reach 100% remission status. Thus, the subject of remission remains under active discussion in the rheumatology community. PMID- 17083756 TI - What should be our treatment goal in rheumatoid arthritis today? AB - Remission should be the treatment aim in management of rheumatoid arthritis (RA) today because joint damage may progress in RA patients with low disease activity but presumably does not progress in patients in clinical remission. However, stringent criteria are needed to define remission status, as some criteria in current use allow for considerable residual disease activity. Even using stringent criteria, remission is achievable in a sizable proportion of patients in clinical trials and practice. Defining remission requires an additional consideration: Should a patient who is receiving medication be regarded as in remission if disease is absent, or must the patient be off treatment to be considered to be in remission? A case is made for aiming for a definition of remission that includes patients who continue medication therapy. PMID- 17083757 TI - ACR remission criteria and response criteria. AB - As additional DMARDs have been added to the armamentarium of rheumatologists over the last 60 years, the approach to the treatment of rheumatoid arthritis has changed. Many clinical studies now are geared toward evaluating the concept of eradicating inflammation as a method to seek the elusive goal of sustained remission in RA. One of the first descriptions of remission in 'RA' was by Short et al in 1948, when he documented the natural progression of the disease. Since that time, various criteria have been developed to define RA remission utilizing clinical, radiographic, and laboratory measures. The most stringent of criteria is the American College of Rheumatology Remission Criteria, developed in 1980, which consists of clinical symptoms and signs of inflammation including fatigue, joint pain, morning stiffness, joint tenderness, joint swelling, and erythrocyte sedimentation rate (ESR). Several reports have compared ACR remission criteria to Disease Activity Score (DAS) values to identify equivalent DAS remission values, and these have been extrapolated to modified versions of the DAS, the Simple Disease Activity Index (SDAI), and Clinical Disease Activity Index (CDAI). The ACR remission criteria and the response measures were not designed for use as the target or goal for the clinical management of individual RA patients in routine clinical practice. Nevertheless, rheumatologists yearn for the eradication of inflammation in all RA patients, and attaining remission may be achievable in the future. PMID- 17083758 TI - Definitions of remission for rheumatoid arthritis and review of selected clinical cohorts and randomised clinical trials for the rate of remission. AB - Various definitions of remission in rheumatoid arthritis (RA) have been proposed. The ACR (American College of Rheumatology--formerly ARA, American Rheumatism Association) remission criteria are strict and include nonspecific symptoms such as fatigue. More recently remission according to the Disease Activity Index (DAS) and DAS28 has been described. However, patients who meet the DAS28 remission cut point of < 2.6 may nonetheless have tender and/or swollen joints. The ACR remission criteria are more rigorous than the requirement of DAS28 <2.6. Newer tools for evaluation of RA activity include the Simplified Disease Activity Index (SDAI) and Clinical Disease Activity Index (CDAI), and cut points for remission according to these new indices have been defined. However, all available remission criteria may ignore important aspects of RA, including physical function and radiographic damage. PMID- 17083759 TI - DAS remission cut points. AB - The Disease Activity Score (DAS) and DAS28 are continuous measures of rheumatoid arthritis (RA) disease activity. Values of DAS %lt;1.6 and DAS28 %lt; 2.6 correspond with an increased likelihood of being in remission. This review presents development of the DAS and DAS28 remission cut points and their interpretation. PMID- 17083761 TI - Remission and radiographic progression in rheumatoid arthritis. AB - Complete remission, defined as the presence of clinical as well as radiographic remission, is the ultimate goal of treatment of rheumatoid arthritis (RA). Functional disability in patients with low disease activity is associated with joint inflammation and joint damage. Despite the methodologic problems of scoring radiographs, studies show that radiographic progression is an important outcome measure, and conventional radiography remains the best available method to assess it. Whether radiographic progression is entirely dependent on the presence of joint inflammation is a matter of debate; some evidence suggests that radiologic progression may continue in patients who appear clinically to be in remission. The potential availability of more effective drugs in the near future presents a need to further define and monitor progression of joint damage by more reliable methods. Better diagnosis of joint damage will assist in our quest to attain and document full remission in RA. Some newer techniques that provide direct assessments of metabolic activity in the inflamed joint appear to predict radiographic progression before it can be detected by conventional methods. Until these techniques are validated and assessed for predictive value, we would advocate that radiographic progression be added to existing criteria for clinical remission, in order to define remission in RA more comprehensively. PMID- 17083760 TI - Can remission be maintained with or without further drug therapy in rheumatoid arthritis? AB - Remission is now the accepted goal of management in rheumatoid arthritis (RA). This article highlights the controversies surrounding the definition of remission and reviews the potential of current treatment options to achieve remission. Defining "true" remission can be difficult based on current criteria, which do not consider structural and physical function. Nonetheless, considerable advances in recent years have made the concept of remission a realistic goal. In early RA, substantial and largely irreversible radiographic damage is seen in 60% of patients within the first 2 years of diagnosis. Early therapeutic intervention would ideally lead to reduction in long-term disability in RA and likelihood of inducing and maintaining remission.Long-term maintenance therapy with disease modifying antirheumatic drugs (DMARDs) has been shown to be effective in preventing flares of disease. Stopping therapy for short periods does not necessarily lead to flares, but the effect on long-term radiographic damage and potential to achieve similar levels of disease control following reinstatement of therapy is not established. Early use of tumour necrosis factor (TNF)-antagonist therapy (e.g. infliximab) has been shown to lead to significant improvement in disease activity measures (clinical and radiologic outcomes) when compared to monotherapy or combination DMARD and corticosteroid therapies. Response was shown to be sustained in 70% of patients receiving TNF-blocking therapy 1 year after stopping treatment. This suggests the significant role of TNF-blocking therapy in enabling sustainable remission without need for long-term administrations, which has important implications for favourable health economics. At present, little published evidence exists on the effects of withdrawal of TNF-blocking therapy in patients with established RA in remission. In conclusion, evidence indicates that remission is a realistic goal, but more evidence is required to establish optimal treatment strategies and define criteria for remission that include imaging and immunological as well as clinical assessment of the disease state. PMID- 17083762 TI - Is remission in rheumatoid arthritis associated with radiographic healing? AB - The precondition for joint damage in rheumatoid arthritis (RA) is inflammation, and the precondition for healing is absence of inflammation. A systematic search for healing phenomena in RA patients in remission has not yet been undertaken. In reports of patients in whom healing was observed, clinical and laboratory data have not been published in part due to space restrictions. However, this preliminary review of the existing literature about repair supports the thesis that a strong association may exist between remission and repair. Several reports indicate that patients in whom radiographic repair was seen were in clinical remission. In most reports clinical response to treatment was very good, and in groups of patients in which scoring was done, evidence of repair was seen in patients with strong inhibition or halt of radiographic progression. In contrast, healing is unlikely to be detected in patients with persistent clinically active disease and/or moderate or strong radiographic progression. In most reports clinical response to treatment was very good, and in groups of patients in which scoring was done, evidence of repair was seen in patients with strong inhibition or halt of radiographic progression. In contrast, healing is unlikely to be detected in patients with persistent clinically active disease and/or moderate or strong radiographic progression. PMID- 17083763 TI - Remission of rheumatoid arthritis: should we care about definitions? AB - A state of remission can be achieved in more and more rheumatoid arthritis (RA) patients. The combination of several RA disease activity measures seems to be important to provide an overall view of disease activity. Remission can be defined by two different approaches: one using a categorical model, requiring criteria for multiple variables to be fulfilled, each with its own threshold value (remission "criteria"); the other using a dimensional model, providing single measures of activity, which allow definition of remission by a single cut point (remission cut points for composite indices). The face validity of remission as defined by composite indices surpasses the one for the "criteria". Likewise, the ones that are not weighted seem to surpass the weighted ones, as can be seen by the significant proportion of patients that continues to have considerable swollen joint counts despite being in Disease Activity Score (DAS) 28 remission. All composite indices seem to perform similarly well as tests for remission using expert judgments as the gold standard. PMID- 17083764 TI - Low disease activity state in rheumatoid arthritis: concepts and derivation of minimal disease activity. AB - With recent advances in therapy, the proportion of patients achieving a satisfactory state of minimal disease activity (MDA) is becoming a more important measure with which to compare different treatment strategies. MDA is between high disease activity and remission and anyone in remission will also be in MDA. This paper summarizes the process of coming to a definition of minimal disease activity in rheumatoid arthritis. Two equivalent preliminary definitions of minimal disease activity for use as secondary outcome measures in clinical trials in RA are proposed: a core-set definition based on the WHO/ILAR core set and a DAS-based definition based on the DAS28. PMID- 17083765 TI - A proposed approach to recognise "near-remission" quantitatively without formal joint counts or laboratory tests: a patient self-report questionnaire routine assessment of patient index data (RAPID) score as a guide to a "continuous quality improvement" s. AB - A proposed approach is presented to recognise a status of "near-remission" in a patient with rheumatoid arthritis (RA) on the basis of patient self-report questionnaire data without formal joint counts or laboratory tests. Indices of patient-reported outcome (PRO) measures distinguish active from control treatments in RA clinical trials at levels similar to American College of Rheumatology (ACR) or disease activity score (DAS) 28 improvement levels. PRO measures on a multidimensional health assessment questionnaire (MDHAQ) can be compiled into a routine assessment of patient index data (RAPID) score. RAPID 3 includes the three PRO measures from the ACR Core Data Set - physical function, pain, and global estimate. RAPID 4 adds a self-report joint count from a rheumatoid arthritis disease activity index (RADAI). RAPID 5 adds a physician estimate of global status. RAPID cores may be classified into four preliminary proposed categories, as "near-remission" (0-1), "low severity" (1.01-2), "moderate severity" (2.01-4), and "high severity" (> 4), analogous to the four categories of the DAS28 of "remission" (< 2.6), as well as "low" (2.6-3.19), "moderate" (3.2-5.1), and "high" (> 5.1) disease activity. RAPID scores are correlated significantly with DAS28 (rho = 0.64-0.67, p < 0.001), and about 75% of patients with DAS < 2.6 have RAPID scores < 2, while about 75% of patients with DAS > 5.1 have RAPID scores > 4. RAPID data are available on one side of one page, and are feasible to collect in standard clinical care. RAPID 3 scores may be calculated in about 10 seconds, and RAPID 4 and RAPID 5 scores in 20 to 30 seconds. RAPID scores every 3 months or more on simple flowsheets can be a basis for a "continuous quality improvement" strategy in standard clinical care to recognise a need for aggressive therapy, an inadequate response to a therapy, and "near- remission" status. PMID- 17083766 TI - Remission as the treatment goal--the FIN-RACo trial. AB - The Finnish Rheumatoid Arthritis Combination Therapy (FIN-RACo) trial is the first rheumatoid arthritis (RA) clinical trial in which remission served as the primary outcome measure. This chapter reviews the philosophical background, study design, and results of the FIN-RACo trial. The study showed that a third of patients with active early RA may achieve remission with a combination of methotrexate (MTX), sulfasalazine (SSZ), hydroxychloroquine (HCQ), and prednisolone. PMID- 17083768 TI - Defining remission in psoriatic arthritis. AB - Driven in part by the introduction of highly effective agents, there has been growing interest in the overall therapeutic approach to patients with psoriatic arthritis (PsA). As with any form of arthritis, the goal of treatment for PsA would be to improve the outcome to the greatest extent possible. In other conditions, such as rheumatoid arthritis, recent discussions have centered on how best to define "remission." For patients with PsA, the heterogeneity among disease manifestations as well as the need to validate outcome measures make definition of remission challenging. In this paper we present a number of key principles and considerations critical to laying the groundwork for defining remission in PsA. PMID- 17083767 TI - Aiming at low disease activity in rheumatoid arthritis with initial combination therapy or initial monotherapy strategies: the BeSt study. AB - AIM: To evaluate the efficacy and safety of four different treatment strategies for patients with early rheumatoid arthritis (RA). METHODS: In the BeSt study, 508 patients with newly diagnosed (< 2 years) active RA were randomised to be treated according to four treatment strategies: 1. sequential monotherapy, 2. step up to combination therapy (both starting with methotrexate), 3. initial combination therapy with methotrexate, sulphasalazine, and a tapered high dose of prednisone, and 4. initial combination therapy with methotrexate and infliximab. Three-monthly therapy adjustments were dictated by calculation of the Disease Activity Score (DAS), with the goal to achieve and maintain a DAS or = 5 kg), child's type extracorporeal circuit vessel and heparin anticoagulation. The replacement solution was delivered pre-dilution after 3 to 4 hours' post-dilution. The blood gas, clinical biochemical items, medium molecule substance (MMS) concentration in blood as well as capillary refill time (CRT), BP, urine output, vasopressors dosage were examined at a set of time points from the beginning to the end of the CVVH. RESULTS: Of the 9 children, 6 had acute renal failure (ARF), 3 had acute respiratory distress syndrome (ARDS), 5 were blood culture positive and all the 9 needed vasopressors to keep BP before CVVH. The blood pH was 7.14 +/- 0.23, base excess (BE) was -11.3 +/- 4.25 mmol/L, MMS was 3532 +/- 519 U/L, PO2/FiO2 was 188 +/- 33, CRT > 5 s, urine output was 0.85 +/- 0.52 ml/(kg.hr) and the adrenalin dosage 1.36 +/- 0.48 microg/(kg.min), and dopamine 16.35 +/- 3.27 microg/(kg.min) before CVVH. The patients' condition was improved much as demonstrated by pH 7.38 +/- 0.16, BE -0.28 +/- 1.37 mmol/L, MMS 2576 +/- 375 U/L, PO2/FiO2 285 +/- 63, CRT < 2 s, and the adrenalin dosage 0.08 +/- 0.04 microg/(kg.min) and dopamine 8.53 +/- 6.72 microg/(kg.min), urine output 2.9 +/- 1.6 ml/(kg.hr) after 24 hour treatment with CVVH. Of the 9 children, 2 died of MODS (1 intussusception complicated with intestine necrosis, 1 severe scald) and 1 was given up because of severe intestinal fistula, the other 6 children recovered at the end. CONCLUSION: CBP was effective in treatment of pediatric septic shock by improving the oxygenation, correcting metabolic acidosis, stabilizing BP, increasing the tissue perfusion and eliminating the medium molecule substances. PMID- 17083782 TI - [Changes of serum neuron specific enolase in rats with septic shock]. AB - OBJECTIVE: To study the changes of serum neuron specific enolase in rats with septic shock. METHODS: The model of septic shock was set up by injection of lipopolysaccharide (LPS, from Escherichia coil O55: B5) at a dose of 25 mg/kg through femoral vein. Twenty male Wistar rats were randomly divided into 2 groups: normal control group (LPS was substituted by same volume of normal saline solution) and septic shock group. Six hours after the septic shock model formed, whole blood was taken for measuring the serum neuron specific enolase (NSE). The brains of the rats were taken for histopathological examination. RESULTS: The serum NSE of septic shock group was significantly higher than that of control group [(10.0781 +/- 0.526) microg/L vs. (3.7188 +/- 0.602) microg/L, P < 0.05]. Neurons were severely damaged 6 hours after injection of LPS. Neuronal necrosis and the damage of blood-brain barrier were seen by light and electron microscope in septic shock group but not in the control group. CONCLUSION: NSE in serum increased when septic encephalopathy occurred, which indicated that NSE might become a marker of neural damage in septic shock. PMID- 17083783 TI - [Unrelated umbilical cord blood stem cell transplantation in a patient with chronic myelocytic leukemia]. PMID- 17083785 TI - [Discussions on the therapeutic approaches to congenital myogenic torticollis]. PMID- 17083784 TI - [Effects of glutamine on matrix metalloproteinase-3 and tissue inhibitor of metalloproteinase-3 expressions in myocardium of rats with sepsis]. AB - OBJECTIVE: The underlying mechanisms for cardiac dysfunction in sepsis include the inhibitory effect of endotoxin and inflammatory factors on myocardium and the decrease in cardiac myocardial cells in number. However, whether there is ventricular remodeling resulted from the abnormalities of extracellular collagen metabolism and whether glutamine (Gln) can protect myocardium from LPS-induced damage as in reperfusion are unknown. The aim of the present study was to examine the effects of Gln on the expressions of matrix metalloproteinase-3 (MMP-3), tissue inhibitor of metalloproteinase-3 (TIMP-3) and their mRNA in myocardium of rats with sepsis. METHODS: Classical rat model of sepsis was established by intraperitoneal injection of lipopolysaccharide (LPS) (4 mg/kg, from Escherichia coli O(55): B(5), Sigma). from 121 Wistar rats aged 18 days were divided into three groups randomly, 0 h control group (normal saline: 1 ml/kg, n = 11), LPS group (LPS: 4 mg/kg, n = 55) and Gln group (LPS: 4 mg/kg and immediately 13.64% glutamine 1 ml/kg, Fresenus, n = 55). Furthermore, LPS and Gln groups were examined at 2 h, 4 h, 6 h, 24 h and 72 h time points (n = 11). On each time point, rats of LPS and Gln groups as well as control group were anesthetized with 1% chloral hydrate injected intraperitoneally at a dosage of 1 ml/kg. Then, rats were sacrificed, and the hearts were isolated. Eight of them were frozen at minus 80 degrees C to measure the expression of TIMP-3 mRNA by using RT-PCR. The expressions of MMP-3 and TIMP-3 were observed with immunohistochemistry and the expression of MMP-3 mRNA was observed by using in situ hybridization. RESULTS: (1) Compared to 0 h, the mRNA expressions of MMP-3 and TIMP-3 in LPS group significantly increased (P < 0.01) with the peak at 6 - 24 h. While, in Gln group, they were significantly higher than those in controls but significantly lower than those in LPS group with the peak at 24 h (P < 0.01). Even at 72 h, they were still higher than those at 0 h (P < 0.05 and P < 0.01). (2) Compared to 0 h, the expressions of MMP-3 and TIMP-3 in LPS group were significantly lower at any other time point with the lowest at 6 h (P < 0.01). In Gln group, these expressions were also significantly lower than those in controls, but significantly higher than those in LPS group with the lowest being postponed to 24 h (P < 0.01). (3) The ultra structure changed obviously. Z line was unclear and the ridge of mitochondrion disappeared. While, in Gln group, the myocardial injury was slight compared to that in LPS group. CONCLUSIONS: MMP-3 mRNA expression was increased and TIMP-3 mRNA expression was depressed in LPS-induced sepsis. Myocardial extracellular matrix was damaged in sepsis. Glutamine might decrease the effects of LPS on MMP-3 and TIMP-3 expressions and postpone the time of myocardial matrix injury. PMID- 17083786 TI - [Summary of consensus meeting on systemic inflammatory response syndrome]. PMID- 17083787 TI - [Summary of the symposium on children, health and law]. PMID- 17083788 TI - [Paroxysmal inspiratory dyspnea as first sign in a patient with nasopharyngeal meningocele]. PMID- 17083789 TI - [Recommended protocol for diagnosis and treatment of septic shock in children]. PMID- 17083790 TI - [Pathogenesis and clinical intervention of systemic inflammatory response syndrome]. PMID- 17083791 TI - [Expression of nuclear factor-kappaB and its inhibitor in alveolar macrophages of patients with neonatal hyaline membrane disease]. AB - OBJECTIVE: Inflammatory reaction and injury in immature lungs are associated with activation of nuclear factor-kappa B (NF-kappaB) to trigger proinflammatory cytokine release, but the mechanism thereof is not fully understood. The present study was conducted to understand possible relationship between expression of NF kappaB and its inhibitor and severity and outcome of neonates with hyaline membrane disease (HMD). METHODS: Serial samples of bronchoalveolar lavage fluid (BALF) were obtained during mechanical ventilation from 31 preterm infants with HMD. These infants were divided into two groups: survivors group [n = 22, birth weight (1500 +/- 320) g and gestational age (31.2 +/- 1.8) weeks] and nonsurvivors group [birth weight (1340 +/- 280) g, gestational age (30.8 +/- 2.1) weeks]. Nineteen preterm infants [birth weight (1470 +/- 280) g, gestational age (30.6 +/- 1.9) weeks] without respiratory disorders were enrolled as control subjects. Alveolar macrophages (AM) were isolated by differential adherence. AM was cultured and treated with lipopolysaccharide (LPS) for 1 hr. Then, nuclear extracts of AM were analyzed by electrophoretic mobility shift assay (EMSA) for NF-kappaB expression. NF-kappaB inhibitor (IkappaB-alpha protein) in cytoplasmic extracts was detected by using Western blotting and IL-1beta and IL-8 in BALF by enzyme-linked immunosorbent assay (ELISA). RESULTS: NF-kappaB complexes were observed by EMSA, they were characterized by competition with cold oligonucleotide and p65-specific antibodies. The addition of an excess of cold oligonucleotide, corresponding to the NF-kappaB binding site, turned off the signal of the band, showing that the band was specific. An excess of an irrelevant oligonucleotide (corresponding to the SP-1) did not show any effect. The addition of an anti-p65 antibody caused the supershift of the two upper bands. After EMSA, the NF-kappaB complexes were quantified by using a ImageQuant software. NF-kappaB expression in AM at 24 hrs was higher in all the patients with HMD as compared with control subjects (survives/control, 34.1 vs 11.4 RDU, P < 0.01; nonsurvivors/control, 55.2 vs 11.4 RDU, P < 0.01). The NF-kappaB expression in AM at 72 hrs was higher than that in control subjects but not for nonsurvivors (survivors/control, 47.8 vs 25.6 RDU, P < 0.01; nonsurvivors/control, 21.8 vs 25.6, P > 0.05). The NF-kappaB expression in AM from nonsurvivors was depressed at 72 hrs as compared to 24 hrs (21.8 vs 55.2, P < 0.01), whereas the NF-kappaB expression in AM from survivors was still higher at 72 hrs than that at 24 hrs (47.8 vs 34.1, t = 4.43, P < 0.01). CONCLUSION: Altered NF-kappaB activation in AM of BALF of neonates with HMD was observed, and it may be mediated by decreased IkappaB synthesis, increased IkappaB degradation, or both. In HMD nonsurvivors NF-kappaB translocation was hampered upon LPS activation. PMID- 17083792 TI - [Effects of different interventional modalities on vascular endothelial dysfunction of obese rats]. AB - OBJECTIVE: To observe the effect of diet-control only, diet-control with swimming training or with polysaccharide sulfate (PSS), a kind of blood lipid-lowering drug on the serum lipid level and vascular endothelial function in obese rats fed by fat-rich-diet. METHODS: Forty Wistar rats were randomly divided into the following 5 groups: group F (n = 8), group N (n = 8), group A (n = 8), group B (n = 8) and group C (n = 8), where the rats were given fat-rich-diet, basic-diet, 12 weeks of diet-control after 8 weeks of fat-rich-diet, 12 weeks of diet-control with swimming training after 8 weeks of fat-rich-diet and 12 weeks of diet control with PSS after 8 weeks of fat-rich-diet, respectively. All rats were sacrificed after 12 weeks of intervention. Then the levels of Lee index, serum total cholesterol (TC), triglyceride (TG), plasma endothelin (ET), nitric oxide (NO) and von Willebrand Factor (vWF) were measured. The protein expression of intercellular adhesion molecule-1 (ICAM-1) in artery endothelium was evaluated by immunohistochemistry (IHC) and the gene expression of ICAM-1 was examined by semi quantitative reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: After the interventions for 12 weeks, the levels of serum TC, TG and ET decreased in group A (P < 0.05). The levels of Lee index, TC, TG, ET, vWF, ICAM-1 protein and ICAM-1 mRNA decreased in group B and C (P < 0.05). Three interventions increased serum NO production (P < 0.05) in group B. CONCLUSIONS: Diet-control could a meliorate the hyperlipidemia and vascular function. Diet-control with swimming training and diet-control with PSS could result in weight loss of rats and meliorate the hyperlipidemia, vascular endothelial function, coagulatory activities and adhesive dysfunction. The effects of diet-control with swimming on vascular endothelial function were prominent. PMID- 17083793 TI - [Interventional approach to the treatment of aneurysms of the perimembranous ventricular septal defects]. AB - OBJECTIVES: To explore applicable protocol for the positioning of ventricular septal defect (VSD) occluder and the selection of the device by retrospective analysis of transcatheter closure approach to the aneurysms of the perimembranous VSD. METHODS: Thirty-five cases of perimembranous VSD with septal aneurysm (19 males and 16 females) from May, 2004 to May, 2005 were included, with a mean age of 5.3 y and mean weight of 17.6 kg. Their angiographic and ultrasound data, and interventional processes were analyzed. Seven segments of the aneurysms were assessed: the diameter of the defect on the left ventricle, the diameter of the defect on the right ventricle, the thickness of ventricular septum, the distance from the farthest end of the aneurysm to the defect, the diameter of the widest part of the aneurysm and the distance between the two farthest orifices on the aneurysm. RESULTS: Sixteen cystiform aneurysms and nineteen tubiform ones were identified with left ventricular angiography. The diameters of the orifices of aneurysms and the diameters of the VSDs ranged from 1.5 mm to 4.1 mm and 2.7 mm to 11.9 mm, separately, with the mean of 2.9 mm and 4.3 mm. From the echocardiography, the distances of the rim of defect to the aortic valve ranged from 2.0 mm to 7.0 mm, with the mean of 4.3 mm. All the interventions were successfully done with symmetrical devices from 4 mm to 14 mm. The left disc of the device was positioned at the defect surface from the left ventricle in 29 cases, and was released at the left side of the orifice in 3 cases. CONCLUSIONS: The positioning of the left disc is mostly determined by the condition for the correct formation of the right disc in the right ventricle after deploying. Generally the defect surface in the left ventricle is most ideal to release the left disc of the device. If the body of aneurysm was too long for the right disc to restore its configuration, the left disc should be released on the left side of the orifice. The selection of device size is determined by the placement of the left disc. When the left disc is to be released at the defect surface in the left ventricle, the device size should be equal to or 1 to 2 mm larger than the diameter of the defect on the left ventricle. When the left disc is to be deployed on the left side of an orifice, the device size should be equal to or 1 mm larger than the defect diameter on the left ventricle when there is a single orifice. In the case of multiple orifices, the minimal size of the device which can cover all the orifices should be selected. PMID- 17083794 TI - [Hypoxia responsive element regulated herpes simplex virus-thymidine kinase system enhances killing effect of gancyclovir on Ewing's sarcoma cell line under hypoxic condition]. AB - OBJECTIVE: To find out a possible approach to improve the effectiveness of radiotherapy and chemotherapy for Ewing's sarcoma by constructing a eukaryotic expression vector expressing herpes simplex virus-thymidine kinase (HSV-TK) regulated by hypoxia responsive element (HRE) under hypoxia and to evaluate the effects of this HRE regulated HSV-TK system on killing effect of gancyclovir (GCV) on Ewing's sarcoma cell line SK-ES under hypoxic condition. METHODS: The HRE was synthesized according to the literature and cloned into the enhancer site of pIRES(2)-EGFP vector to obtain the pHRE recombinant plasmid. The HSV-TK was amplified by PCR and cloned into the multiple clone site of pIRES(2)-EGFP and pHRE to obtain pTK and pHRE-TK recombinant plasmid. The human Ewing's sarcoma cell line SK-ES was transfected by pTK or pHRE-TK recombinant plasmid with liposome and then was exposed to normoxic (21% oxygen) or hypoxic (3% oxygen) condition. The expression of enhanced green fluorescent protein (EGFP) was monitored by fluorescent microscopy. The sensitivity of human Ewing's sarcoma cell line SK-ES transfected with pTK or pHRE-TK recombinant plasmid to the anti tumour drug GCV was determined with the method of tetrazolium (MTT) after treating with GCV for five days. RESULTS: (1) The result of sequencing showed that the recombinant plasmid pHRE contained HRE, and that the recombinant plasmid pTK and pHRE-TK contained HSV-TK gene in the sense direction. (2) Comparison of fluorescent optical density (FOD) showed that (1) the EGFP FOD value of pHRE and pHRE-TK group cells exposed to hypoxia was significantly higher than those exposed to normoxia (P < 0.01); (2) when the cells were exposed to hypoxia, the EGFP FOD value of pHRE and pHRE-TK group cells was significantly higher than that of pTK and empty vector group (P < 0.01); (3) there was no significant difference among the four groups of cells when they were exposed to normoxia (P > 0.05). (3) Comparison of the sensitivity of four groups of cells to GCV showed that (1) the cells in pHRE-TK and pTK groups were much more sensitive to GCV than the cells in pHRE group under hypoxia condition (P < 0.01), the higher the GCV concentration, the greater the difference; (2) the cells of pHRE-TK group were more sensitive to GCV than those in pTK group under hypoxic condition (P < 0.01), but was almost equally sensitive under normoxic condition (P > 0.05); (3) the pHRE-TK group cells had higher sensitivity to GCV under hypoxia than normoxia (P < 0.01) while the pTK group cells had almost the same sensitivity to GCV under hypoxia and normoxia (P > 0.05). CONCLUSION: (1) The eukaryotic expression vector expressing herpes simplex virus-thymidine kinase (HSV-TK) regulated by hypoxia responsive element (HRE) under hypoxia was constructed successfully. (2) HRE could up regulate expression of EGFP by SK-ES cells under hypoxia condition. (3) HRE could enhance the killing effect of HSV-TK/GCV system on human Ewing's sarcoma cell line SK-ES under hypoxic condition. PMID- 17083795 TI - [Expression and role of TLR and SOCS mRNA in newborn infants]. PMID- 17083796 TI - [Effect of noise on auditory system development of newborn infants treated in the neonatal intensive care unit]. PMID- 17083797 TI - [Review of prior research in China: child abuse and its impact on the victims]. PMID- 17083798 TI - [Antisense gene therapy of respiratory syncytial virus infection]. PMID- 17083799 TI - [Airway remodeling and asthma]. PMID- 17083800 TI - [Hydrops fetalis due to placental chorioangioma: report of a case]. PMID- 17083801 TI - [A brief description of the 6th Asian Regional Conference on Prevention of Child Abuse and Neglect]. PMID- 17083802 TI - [Upper gastrointestinal bleeding in a case with Dieulafoy lesion]. PMID- 17083803 TI - [Predictive value of serum soluble fms-like tyrosine kinase 1 concentration in preeclampsia at second trimester]. AB - OBJECTIVE: To explore the predictive value of serum soluble fms-like tyrosine kinase-1 (sFlt-1) and vascular endothelia growth factor (VEGF) levels in preeclampsia at second trimester. METHODS: Serum sFlt-1, VEGF concentrations were determined in 172 initial normal pregnant women at 26 - 28 gestation week. The outcomes of pregnancies were followed. In a cohort of 172 pregnant women, 16 cases of preeclampsia were developed (preeclampsia group), and 156 cases were with no complication (control group). RESULTS: The serum levels of sFlt-1 in preeclampsia group (11.4 +/- 6.2) microg/L were significantly higher than that in control group (4.5 +/- 2.1) microg/L (P < 0.01). The serum levels of sFlt-1 in precelampsia women with the onset before 32 gestation week and fetal growth retardation, (14.0 +/- 6.8) microg/L, (14.4 +/- 6.7) microg/L were significantly higher than that in women with the onset after 32 gestation week and with no fetal growth retardation (9.0 +/- 4.1) microg/L, (8.9 +/- 4.0) microg/L, respectively (P < 0.05). There was no significant difference in the serum levels of VEGF between preeclampsia group and control group. A sFlt-1 cutoff value of 8.75 microg/L at 26 - 28 gestation week yielded a sensitivity of 87.5%, specificity of 97.4%, positive predictive value of 80.0%, negative predictive value of 88.5%, respectively, for subsequent onset of preeclampsia. CONCLUSION: Maternal serum sFlt-1 concentration at second trimester can be used as an early predictive marker of preeclampsia. PMID- 17083804 TI - [Differential expression of extracellular matrix metalloproteinase inducer in normal placenta and preeclampsia placenta]. AB - OBJECTIVE: To study the expression of extracellular matrix metalloproteinase inducer (EMMPRIN) in preeclampsia placenta and the relation with preeclampsia attacks. METHODS: Forty-four samples from pregnant women with preeclampsia (preeclampsia group), 38 samples from pregnant women with eclampsia, and 49 samples from normal pregnancies (control group) were obtained. We detected the expression of EMMPRIN in placenta by immunohistochemistry and the expression of EMMPRIN mRNA by RT-PCR. RESULTS: (1) EMMPRIN positive expression: in preeclampsia group, the moderate expression rate was 18% (8/44) and the strong positive rate was 9% (4/44); in eclampsia group moderate positive rate was 21% (8/38) and strong positive rate 13% (5/38). The difference of the two groups was insignificant (P > 0.05). In control group the moderate positive rate was 12% (6/49) and strong positive rate 82% (40/49), the difference from the preeclampsia and the eclampsia groups was significant (P < 0.001). (2) EMMPRIN mRNA expression: in preeclampsia group EMMPRIN mRNA expression in term placenta (37 - 40 gestational weeks) was 0.342 +/- 0.002, and in eclampsia group 0.344 +/- 0.023; the difference between the two groups was insignificant (P > 0.05). In control group EMMPRIN mRNA expression in term placenta (37 - 40 gestational weeks) was 0.872 +/- 0.094, the differences between the control group and preeclampsia and eclampsia groups were both significant (P < 0.001). CONCLUSION: The decrease in the expression of EMMPRIN in placenta is an important cause of preeclampsia onset; expression rate of EMMPRIN may serve as an indicator in predicting preeclampsia. PMID- 17083805 TI - [Expression of hypoxia-inducible factor-1alpha, vascular endothelial growth factor and sFlt-1 in preeclampsia placenta]. AB - OBJECTIVE: To investigate the expression and correlation of hypoxia-inducible factor-1alpha (HIF-1alpha), vascular endothelial growth factor (VEGF) and sFlt-1 in the preeclampsia placenta, and discuss their significance in the pathogenesis of preeclampsia. METHODS: Placentas were collected from 20 pregnant women with preeclampsia as study group and 15 normal pregnant women as control group. The expressions of HIF-1alpha, VEGF and sFlt-1 protein were semi-quantitatively analyzed with immunohistochemical assay and mRNA level was determined using reverse transcription polymerase chain reaction (RT-PCR) technique. RESULTS: (1) the expression of HIF-1alpha and sFlt-1 protein in preeclampsia group obviously increased. Strong (+++) positive expression was observed in 9 and 11 cases respectively, significantly higher than in control group (2 and 3 cases) (P < 0.05), however, VEGF expression obviously reduced in preeclampsia group (P < 0.01). (2) the level of HIF-1alpha and sFlt-1 mRNA in preeclamptic placenta was 0.604 +/- 0.013, 0.898 +/- 0.041, significantly higher than 0.208 +/- 0.007 and 0.559 +/- 0.244 in normal placenta (P < 0.05). Although the level of VEGF mRNA increased in preeclampsia placenta, it was not significantly different from that in normal placenta (P > 0.05). The ratio of VEGF mRNA/sFlt-1mRNA obviously reduced in preeclampsia group and was significantly lower than in control group (P < 0.05). (3) in preeclampsia group, HIF-1alpha mRNA expression was positively correlated with the expression of sFlt-1 mRNA (r = 0.577, P < 0.05), and negatively correlated with the ratio of VEGF mRNA/sFlt-1 mRNA (r = -0.376, P < 0.05). CONCLUSION: Abnormal high HIF-1alpha expression in preeclampsia placenta indicates that HIF-1alpha might play an important role in the pathogenesis of preeclampsia, possibly through affecting the cytotrophoblastic invasion and placental vascular reconstruction via the modulation of VEGF and sFlt-1 gene transcription. PMID- 17083806 TI - [Study of fetal lymphocyte of intrahepatic cholestasis of pregnancy]. AB - OBJECTIVE: To explore effect of fetal lymphocyte on pathogenesis of intrahepatic cholestasis of pregnancy (ICP). METHODS: Twenty pregnant women with ICP and 20 normal pregnant women were enrolled in the study. The single mixed lymphocyte culture/reaction (MLC/MLR) was conducted using inactive lymphocyte obtained from maternal peripheral blood and lymphocyte of cord blood from fetus. Antigen induced-lymphocyte-proliferation-reaction was used for dermic soluble antigen and decidual soluble antigen obtained from maternal blood and cord blood from fetus. The intense of proliferation was calculated and compared between normal and ICP complicated pregnancies. RESULTS: (1) The level of intense of proliferation of fetal lymphocyte was significantly increased in ICP group 2.75 +/- 0.36 than those of normal control group 1.45 +/- 0.19 in single mixed lymphocyte culture (P < 0.05). (2) The level of intense of proliferation of fetal lymphocyte was significantly increased in ICP group 1.45 +/- 0.19 than those of normal control group 0.67 +/- 0.24 in decidual soluble antigen induced lymphocyte proliferation reaction (P < 0.05). (3) The level of intense of proliferation of fetal lymphocyte was significantly increased in ICP group (1.22 +/- 0.44) than those of normal control group (0.66 +/- 0.27) in dermic soluble antigen induced lymphocyte proliferation reaction. CONCLUSIONS: (1) The fetal lymphocyte may be one of the effector cells in pathogenesis of ICP. (2) The disturbance of fatal-maternal immune-tolerance is one of the important mechanisms underlying ICP. PMID- 17083808 TI - [Clinical analysis of fallopian tube prolapse after hysterectomy]. AB - OBJECTIVE: To investigate the clinical diagnosis, treatment and prevention of fallopian tube prolapse (FTP) after hysterectomy. METHODS: A total of 7949 patients received hysterectomy from 1983 to Aug 2005 in Peking Union Medical College Hospital, including 6229 cases of trans-abdominal hysterectomy (TAH), 780 cases of transvaginal hysterectomy (TVH), and 940 cases of laparoscopic assisted vaginal hysterectomy (LAVH). Nine cases (including 1 case from other hospital) of FTP after hysterectomy were analyzed retrospectively for their symptoms, diagnosis and treatment. All of them were diagnosed according to the results of histology and follow-up. RESULTS: The overall incidence of FTP after hysterectomy was 0.11% (9/7949). Incidence of FTP after trans-abdominal hysterectomy was 0.08% (5/6229), after vaginal hysterectomy 0.51% (4/780), and after laparoscopic assisted vaginal hysterectomy 0 (0/940). There were no symptoms in 3 cases, but the other 6 cases had symptoms. The pelvic examination revealed the typical prolapsed fimbrial end of a fallopian tube in 3 cases and red granulation tissue in the other 6 cases. All of them were excised vaginally and cauterized. The results were confirmed by histological examination. No recurrent cases were reported in follow up. CONCLUSIONS: FTP is a rare complication after hysterectomy. The prognosis is well after proper diagnosis and treatment. Salpingectomy or fixation of accessories into the pelvic wall and complete peritonealisation at the time of hysterectomy are important methods to prevent FTP after hysterectomy. PMID- 17083809 TI - [Study on causes and treatment of repeated vulvovaginitis in girlhood]. AB - OBJECTIVE: To explore the causes and treatment of repeated vulvovaginitis in girlhood in order to improve its prevention and treatment. METHODS: Fifty-one girls with repeated vulvovaginitis (age < or = 10 years) admitted to The Second Affiliated Hospital of Sun Yat-sen University from Jan. 1990 to Nov. 2004 were reviewed retrospectively. RESULTS: We found 28 girls (55%) suffering from non specific vulvovaginitis and 14 ones (27%) suffering from posterior recto-vaginal fistula with in 51 patients. Five girls (10%) were smitten with vulval ulcer and 3 ones (6%) had been were found with vaginal foreign bodies. One girl (2%) was smitten with adhesion of labia minora. The vaginal discharges taken from 21 girls were cultured. Seventeen cases found bacteria. The positive rate of bacteria culture in the 21 cases reached 81%, in which, E.coli accounted for 5 cases (24%), staphylococcus and streptococcus accounted for 3 cases (14%) respectively. Patients suffering from non-specific vulvovaginitis and vulval ulcer accepted external lotion, antibiotic ointment or combining with antibiotics. Patients suffering from posterior recto-vaginal fistula accepted fistulectomy. Three girls who found vaginal foreign bodies took out of foreign bodies by hysteroscope. Fifty-one girls all were cured after appropriate therapy. CONCLUSIONS: Vulvovaginitis is the most common gynecologic diagnosis in girlhood. The principal cause of repeated invasion is non-specific vulvovaginitis and the secondly one is posterior recto-vaginal fistula. It need overhaul during the diagnosis. It is very availability to use hysteroscopy and do bacteria culture + antibiotic sensitivity test for repeated pediatric vulvovaginitis. PMID- 17083811 TI - [Construction and utilization of the prognostic model of serous ovarian adenocarcinoma]. AB - OBJECTIVE: To analyze the related factors with prognosis in patients with serous ovarian adenocarcinoma and to set up a prognostic model of serous ovarian adenocarcinoma. METHODS: The clinical, pathological and follow-up data of 104 cases with serous ovarian adenocarcinoma were retrospectively analyzed. Kaplan meier univariate analysis was used to screen the prognostic factors; COX univariate and multivariate analyses were used to determine the risk coefficient of each factors and different layers in each factor. Pearson rank correlation was used to reject the influence of different factors with each other. And the prognostic model of serous ovarian adenocarcinoma was set up based on the result of the above study, which could be used to deduce the survival probability of patients with serous ovarian adenocarcinoma. RESULTS: International Federation of Gynecology and Obstetrics (FIGO) stage (P = 0.0029), histological grade (P = 0.0054), residual disease (P = 0.0000), metastasis of lymph nodes (P = 0.0000) and chemotherapy (P = 0.0000) were the related factors of prognosis in patients with serous ovarian adenocarcinoma, of which FIGO stage was the most important one, followed sequentially by histological grade, metastasis of lymph node, residual disease and chemotherapy (the independent risk coefficient of each factor was 1.3392, 0.9206, 0.7071, 0.6004, 0.4985 in sequence). We set up a prognosis model according to the prognostic index of each factors. The effect of chemotherapy and residual disease on prognosis could be quantified by this model, and the higher the score, the lower the survival probability of patients. CONCLUSIONS: FIGO stage, histological grade, residual disease, metastasis of lymph nodes and chemotherapy are important prognostic factors of serous ovarian adenocarcinoma. This model can be used to estimate the prognosis of patients with serous ovarian adenocarcinoma, and the effect of both chemotherapy and residual disease on the prognosis could be quantified by the model. PMID- 17083810 TI - [Detection and significance of phosphatidylinositol 3-kinase in adipose tissue of polycystic ovary syndrome patients with insulin resistance]. AB - OBJECTIVE: To investigate the expression of phosphatidylinositol 3-kinase (PI-3K) in adipose tissue of polycystic ovary syndrome patients (PCOS), and explore molecular mechanisms of insulin resistance (IR) in PCOS. METHODS: Samples from patients with PCOS with IR (n = 19), PCOS without IR (n = 10) and controls (n = 15) were collected. Serum fasting insulin (FIN) and fasting plasma glucose (FPG) were measured. Insulin resistance index was calculated using homeostasis model assessment (HOMA) to analyze the relationship between these markers and IR. Western blot technique was used to detect the PI-3K p85 subunit. Gene expression of PI-3K p85 subunit was detected by reverse transcription polymerase chain reaction (RT-PCR) method. Kinase activity was detected by immunoprecipitation, thin-layer chromatography and gamma scintillation counting. RESULTS: (1) The levels of FIN [(25.2 +/- 3.8) mU/L] and HOMA-IR (1.6 +/- 0.3) in PCOS with IR were significantly higher than those in PCOS without IR [(13.4 +/- 3.8) mU/L, 0.9 +/- 0.3] and controls [(9.5 +/- 2.6) mU/L, 0.5 +/- 0.3; all P < 0.05). (2) There was no significant difference in the protein (0.65 +/- 0.10) and gene expression (0.92 +/- 0.12) of PI-3K p85 subunit in PCOS with IR compared with PCOS without IR (0.72 +/- 0.10, 1.01 +/- 0.10) and control groups (0.73 +/- 0.14, 1.00 +/- 0.12; P > 0.05). (3) PI-3K activity in PCOS with IR (81%) and PCOS without IR (89%) was significantly decreased (P < 0.01, P < 0.05) and negatively correlated with HOMA-IR (r = -0.69, P < 0.01; r = -0.62, P < 0.05). CONCLUSIONS: No significant difference in the protein and gene expression of PI-3K p85 subunit in PCOS with IR is found. The decreased PI-3K activity may lead to IR of PCOS. PMID- 17083812 TI - [Feasibility study of laparoscopic complicate myomectomy: analysis of 67 cases]. AB - OBJECTIVE: To investigate the feasibility of complicate myomectomy. METHODS: Six seven patients with complicated uterine myomas undergoing laparoscopic myomectomy were retrospectively analyzed. The myomectomy was done using ureteral infravision imaging system or/and with self-made myoma segregate-stick. Among the total, there are 29 cases of multiple myomas (the number of myomas > or = 5) and 23 cases of single myoma (the diameter of myoma > or = 7cm, including 19 cases of intramural myoma, 4 cases of subserous myoma), 6 cases of myoma of broad ligament of uterus and 9 cases of cervical myoma. RESULTS: All cases were performed successfully laparoscopically. No intra-operative laparotomy or complications occurred. The average operating time and blood loss were (114 +/- 32) min and (114 +/- 78) ml respectively. The average time of hospital stay was 5.1 d. The average operating time and blood loss in the group (including multiple myoma group, intramural myoma group and cervical myoma) were significantly exceeded the other groups (including myoma of broad ligament of uterus group and subserous myoma group). CONCLUSIONS: Laparoscopic complicate myomectomy can be performed and the operation indication is enlarged using Ureteral Infravision Imaging System. Advancement in surgical instruments and expert operating skills are the key to operation success. PMID- 17083813 TI - [Levonorgestrel-releasing intrauterine system for treatment of dysmenorrhea associated with adenomyosis]. AB - OBJECTIVE: To evaluate the efficacy and safety of a levonorgestrel-releasing intrauterine system (LNG-IUS) for the treatment of dysmenorrhea associated with adenomyosis. METHODS: We recruited 48 women with moderate or severe dysmenorrhea associated with adenomyosis. All women were inserted of LNG-IUS into their uterine cavity from days 5 - 7 of their periods and maintained for 12 months. We compared the visual analogue scale (VAS) scores and verbal rating scale (VRS) scores of their dysmenorrhea and dyspareunia at baseline and 12 months follow-up. RESULTS: Forty-four women completed the study. There were significant differences between mean VAS and VRS scores changes of dysmenorrhea and dyspareunia at baseline and 12 months follow-up, those of dysmenorrhea dropping from 75 +/- 13 to 11 +/- 11 and 2.3 +/- 0.4 to 0.4 +/- 0.3, those of dyspareunia dropping from 54 +/- 19 to 4 +/- 4 and from 1.6 +/- 0.8 to 0.2 +/- 0.2 respectively. Overall 29 women (66%) were very satisfied or satisfied with the one-year treatment. CONCLUSION: Insertion of LNG-IUS alleviates moderate or severe dysmenorrhea associated with adenomyosis remarkably. PMID- 17083814 TI - [Preliminary study of membrane estrogen receptor expression in endometrial carcinoma cells]. AB - OBJECTIVE: To analyze if there are membrane estrogen receptors (ER) in endometrial carcinoma and if there is some relationship between membrane ER and nuclear ER. METHODS: The cell membrane and total cell ERalpha and ERbeta expressions of high and moderate differentiation endometrial carcinoma cells (Ishikawa and HEC-1A cells) were analyzed. Intermittent immunofluorescence dyeing and fluorescent microscopy were carried out with the cells treated with polyformaldehyde and Triton X-100. Intermittent immunofluorescence dyeing and flow cytometry were carried out with the live cells and the cells treated with Triton X-100 respectively. RESULTS: There were fluorescences on the membrane of the Ishikawa and HEC-1A cells which were treated with polyformaldehyde. When the cells were treated with Triton X-100, the fluorescences were also seen inside the cells. The fluorescence intensity of ERalpha and ERbeta in Ishikawa cell membrane (1.09 +/- 0.21, 1.27 +/- 0.33) was stronger than the control, but there were no significant differences (P > 0.05). When treated with Triton X-100, the total cell fluorescence intensity of ERalpha and ERbeta in Ishikawa cell (4.21 +/- 0.34, 4.69 +/- 1.96) was stronger than the membrane (P < 0.05). The ERalpha and ERbeta fluorescence intensity of HEC-1A cell membrane (1.58 +/- 0.13, 1.49 +/- 0.04) were stronger than the control (P < 0.05). The fluorescence intensity of ERalpha and ERbeta of the HEC-1A cell (2.34 +/- 0.33, 2.52 +/- 0.15) was stronger than the membrane also (P < 0.05). The membrane ERalpha fluorescence intensity of Ishikawa was lower than HEC-1A (P = 0.028). But the total cell ERalpha fluorescence intensity of Ishikawa was higher than HEC-1A (P = 0.002). CONCLUSIONS: There are membrane ER on endometrial carcinoma cells Ishikawa and HEC-1A. The membrane ER must have some similarity to the nuclear receptor. There is no direct correlation between the quantity of the membrane ER and nuclear ER. PMID- 17083830 TI - [Pathophysiology of preeclampsia-eclampsia]. PMID- 17083831 TI - [Expression of trophoblast invasion related genes mRNA and protein in human placenta in preeclampsia]. AB - OBJECTIVE: To study the mRNA and protein expression of matrix metalloproteinase (MMP) 9, 2, metastasis suppressor gene KiSS-1, tissue inhibitor of metalloproteinase-1 (TIMP-1) and TIMP-2 in placenta of preeclampsia patients and their relation to the pathogenesis of preeclampsia. METHODS: Expression of MMP-9, MMP-2, KiSS-1, TIMP-1, TIMP-2 mRNAs and proteins in placenta from 27 cases of preeclampsia, 10 cases of gestational hypertension and 30 cases of normal term pregnant women was detected by semi-quantitative RT-PCR, western blotting, and enzyme activity of MMP-9 and MMP-2 was measured by gelatin zymography. RESULTS: (1) Expression of MMP-9 and MMP-2 mRNA in placenta of preeclampsia (0.39 +/- 0.05 and 0.71 +/- 0.16) was significantly lower than that in normal term pregnancy (0.78 +/- 0.11 and 1.63 +/- 0.31, P < 0.05). Expression of KiSS-1 and TIMP-1 mRNAs in placenta of preeclampsia (1.97 +/- 0.21 and 1.11 +/- 0.18) was significantly higher than that in normal term pregnancy (0.69 +/- 0.27 and 0.65 +/- 0.19) (P < 0.05.). There was no significant difference in TIMP-2 mRNA level between preeclampsia and normal pregnancy (P > 0.05). (2) Expression of MMP-9 and MMP-2 proteins in preeclampsia (1.07 +/- 0.35 and 0.74 +/- 0.23) was significantly lower than that in normal term pregnancy (2.43 +/- 0.92 and 1.48 +/ 0.78) (P < 0.05). Expression of KiSS-1 and TIMP-1 proteins in placenta of preeclampsia (2.46 +/- 0.39 and 1.51 +/- 0.40) was significantly higher than that in normal pregnancy (0.91 +/- 0.35 and 0.93 +/- 0.56) (P < 0.05). There was no significant difference in TIMP-2 protein level between preeclampsia and normal pregnancy (P > 0.05). (3) Enzyme activity of MMP-9 and MMP-2 in placenta of preeclampsia [(2.67 +/- 0.53) gray level.g(-1).L(-1) and (1.13 +/- 0.28) gray level.g(-1).L(-1))] was significantly lower than that in placenta of normal pregnancy [(8.44 +/- 3.70) gray level.g(-1).L(-1) and (3.87 +/- 1.43) gray level.g(-1).L(-1)] (P < 0.05). CONCLUSION: Abnormal expression of MMP-9, MMP-2, KiSS-1 and TIMP-1 can cause insufficiency invasion of trophoblast in preeclampsia and superficial placentation, which may play an important role in the pathogenesis and development of preeclampsia. PMID- 17083832 TI - [Expression of transforming growth factor-beta 1, vascular cell adhesion molecule 1 and endothelium-selectin in placenta of patients with pre-eclampsia]. AB - OBJECTIVE: To study the change and significance of the expression of transforming growth factor-beta 1 (TGF-beta1), vascular cell adhesion molecule-1 (VCAM-1) and endothelium-selectin (E-selectin) in placenta of patients with pre-eclampsia. METHODS: Twenty normal pregnant women (control group) and 40 women with pre eclampsia (pre-eclampsia group, including 16 women with mild pre-eclampsia and 24 women with severe pre-eclampsia) were selected. The cellular distribution of TGF beta1, VCAM-1 and E-selectin in placenta in both groups was determined by immunohistochemistry, and the mean density was measured by computer image analysis system. RESULTS: (1) The level of TGF-beta1 in placental villous syncytiotrophoblast of pre-eclampsia group (70.7 +/- 0.5) was significantly higher than that of control group (70.3 +/- 0.6), while the level of VCAM-1 and E selectin in pre-eclampsia group (VCAM-1: 82.5 +/- 0.5, E-selectin: 53.5 +/- 0.5) was significantly lower than that of control group (VCAM-1: 82.8 +/- 0.3, E selectin: 53.8 +/- 0.4) (P < 0.05). However, there were no significant differences in women with mild pre-eclampsia (TGF-beta1: 70.6 +/- 0.6, VCAM-1: 82.4 +/- 0.6, E-selectin: 53.4 +/- 0.5) and severe ones (TGF-beta1: 70.8 +/- 0.4, VCAM-1: 82.6 +/- 0.5, E-selectin: 53.6 +/- 0.5) (P > 0.05); (2) The level of E selectin in placental villous capillary endothelial cells of pre-eclampsia group (63.0 +/- 0.5) was significantly higher than that of control group (62.6 +/- 0.4) (P < 0.05), while there was no significant difference in women with mild pre eclampsia (63.2 +/- 0.4) and severe pre-eclampsia (62.9 +/- 0.5) (P > 0.05). CONCLUSION: TGF-beta1, VCAM-1 and E-selectin are not only related to placenta shallow bed of pre-eclampsia, but also participate in pathogenic process of vascular endothelial damage of pre-eclampsia. PMID- 17083833 TI - [Changes of maternal plasma and umbilical cord plasma inhibin and epidermal growth factor in patients with hypertensive disorder complicating pregnancy]. AB - OBJECTIVE: To study the levels of inhibin (INH) and epidermal growth factor (EGF) in maternal plasma and umbilical cord plasma of patients with hypertensive disorder complicating pregnancy, and to explore their influence on the disease and fetal growth. METHODS: Enzyme linked immunosorbent assay (ELISA) was used to detect maternal and umbilical cord plasma INH and EGF levels in 65 patients with hypertensive disorder complicating pregnancy (test groups) and 21 normal pregnant women (control group). RESULTS: Plasma level of INH in test groups (499 +/- 52) ng/L was significantly higher than that (421 +/- 36) ng/L in control group (P < 0.01); however, the umbilical cord plasma level of INH had no significant difference (P > 0.05). Plasma level of EGF in test groups (408 +/- 60) ng/L was significantly lower than that (463 +/- 87) ng/L in control group (P < 0.05), also there was significant difference in umbilical cord plasma level of two groups (232 +/- 99) ng/L vs (380 +/- 97) ng/L (P < 0.01). The level of EGF in umbilical cord blood was positively correlated with newborn's body weight and placental weight. CONCLUSIONS: Plasma levels of INH and EGF in pregnancy women are related with hypertensive disorder complicating pregnancy. EGF level of umbilical cord blood affects the growth of fetus and placenta. PMID- 17083834 TI - [Expression and significance of soluble fms-like tyrosine kinase 1 in preeclampsia placenta]. AB - OBJECTIVE: To investigate the expression and significance of serum soluble fms like tyrosine kinase 1 (sFlt-1) in preeclampsia placenta. METHODS: Immunohistochemistry and semi-quantitative RT-PCR were used to investigate the expression of sFlt-1 and sFlt-1 mRNA in placenta of 30 women with preeclampsia (including mild and moderate preeclampsia 11 cases, severe preeclampsia 19 cases) and 45 normal pregnant women (including first trimester pregnancy 18 cases, 2nd trimester pregnancy 12 cases, and 3rd trimester pregnancy 15 cases). The levels of vascular endothelial growth factor (VEGF) and sFlt-1 in serum were detected by enzyme linked immunosorbent assay. RESULTS: (1) The expression of sFlt-1 mRNA was significantly higher in preeclampsia group (0.90 +/- 0.11) compared with the third trimester group (0.80 +/- 0.06; P < 0.01), and was higher in the severe preeclampsia group (0.93 +/- 0.12) than that in the mild group (0.85 +/- 0.05; P < 0.05). (2) The mean density of sFlt-1 in preeclampsia placenta (0.156 +/- 0.008) was significantly higher than that in the third trimester group (0.143 +/- 0.009, P < 0.01), and was higher in the severe group (0.159 +/- 0.008) than in the mild group (0.151 +/- 0.005; P < 0.05). (3) The level of VEGF in serum of the preeclampsia group (19.3 +/- 2.9) ng/L was significantly lower than that of the third trimester group [(30.2 +/- 3.1) ng/L, P < 0.01]. The level of sFlt-1 in serum of the preeclampsia group (30.2 +/- 13.7) microg/L was significantly higher than that of the third trimester group [(7.4 +/- 3.1) microg/L, P < 0.01]. (4) There was significant correlation between the serum level of sFlt-1 and the expression of sFlt-1 mRNA or sFlt-1 in placenta of normal pregnancy group (r = 0.314, P < 0.05; r = 0.439, P < 0.01) and preeclampsia groups (r = 0.372, r = 0.383; P < 0.05). CONCLUSION: Upregulation of sFlt-1mRNA and excess expression of sFlt-1 in placenta may induce higher level of sFlt-1 in serum, which may be involved in the pathophysiological processes of preeclampsia. PMID- 17083835 TI - [Effect of magnesium sulfate on fetal rats of fetal growth retardation and its relation with expression of caspase-3 on the placenta of maternal rat]. AB - OBJECTIVE: To investigate the effect of magnesium sulfate on the fetal rats of fetal growth retardation (FGR) and its relation with expression of caspase-3 on the placenta of maternal rat. METHODS: Model of FGR was constructed according to the method of passive smoking. The pregnant rats were divided into control group (n = 10), therapy group (n = 18) and FGR group (n = 10). The therapy group rats were given different doses of magnesium sulfate by subcutaneous injection: low dose group (300 mg/kg, n = 10), high dose group (600 mg/kg, n = 8). Serum concentration of magnesium sulfate was monitored. The expression of caspase-3 was measured by immunohistochemistry method and RT-PCR. RESULTS: Both of the concentration of magnesium sulfate in high and low dose groups (0.72 +/- 0.13), (0.61 +/- 0.03) mmol/L were higher than the FGR group (0.55 +/- 0.03) mmol/L (P < 0.01); the weight of placenta and fetal rat in high dose group [(0.80 +/- 0.16) and (3.58 +/- 0.10) g] were more than those of FGR group [(0.63 +/- 0.05) and (2.95 +/- 0.46) g] (P < 0.05, P < 0.01); the expression of mRNA and protein of caspase-3 in high dose group [(0.361 +/- 0.030), (183.0 +/- 3.3)] was lower than the FGR group [(0.626 +/- 0.036), (199.5 +/- 4.7)] (P < 0.05); the expression of mRNA in low dose group (0.525 +/- 0.029) was higher than the high dose group (P < 0.05); serum concentration of magnesium sulfate of maternal rat was correlated with the weight of fetal rat (r = 0.899, P = 0.038) and the expression of mRNA and protein of caspase-3 in placenta (r = -0.747, P = 0.033; r = -0.915, P = 0.001). CONCLUSION: It is suggested that magnesium sulfate increases the weight of fetal rat probably by depressing the expression of caspase-3 of placenta, which improves the placental function. PMID- 17083836 TI - [Thyroid function and thyroid autoimmunity at the late pregnancy: data from 664 pregnant women]. AB - OBJECTIVE: To study the prevalence of thyroid diseases, as well as characteristics of the disease spectrum and thyroid autoimmunity in women at the end of pregnancy. METHODS: Six hundred and sixty-four pregnant women (pregnancy group) and 276 non-pregnant women (control group) were enrolled in the study. Serum thyrotropin (TSH), thyroid peroxidase antibody (TPOAb), free T(3) (FT(3)) and free T(4) (FT(4)) were measured by high-sensitive immunochemiluminescent assay, and urinary iodine was also examined at the end of pregnancy. Overt hyperthyroidism was diagnosed when both TSH < 0.3 mU/L and FT(4) and/or FT(3) levels were elevated. Subclinical hyperthyroidism was diagnosed when TSH < 0.3 mU/L with normal FT(4) and FT(3) levels. The diagnostic criteria for overt hypothyroidism was TSH > 4.8 mU/L accompanied by decreased FT(4), and for subclinical hypothyroidism was TSH > 4.8 mU/L with normal FT(4) and FT(3) levels. RESULTS: (1) The median urinary iodine (MUI) of pregnancy group was 201.5 microg/L, and that of control group was 196.0 microg/L (P > 0.05). Women in the two groups were iodine-adequate. (2) The overall prevalence of thyroid diseases in pregnancy group and control group was 7.8% (52/664) and 6.9% (19/276), respectively (P > 0.05). (3) As for the diseases pattern, there were obvious differences between the two groups. In pregnancy group, the prevalence of hyperthyroidism was lower than that of hypothyroidism (1.1% vs 6.8%, P < 0.01). In control group, the prevalence of hyperthyroidism and hypothyroidism was 4.7% and 2.2%, respectively (P > 0.05). Compared with control group, the prevalence of hyperthyroidism in pregnancy group was much lower (1.1% vs 4.7%, P < 0.01), mainly due to the decrease of overt hyperthyroidism; whereas, the increment of subclinical hypothyroidism resulted in the higher prevalence of hypothyroidism in pregnancy group (6.8% vs 2.2%, P = 0.01). (4) The median TSH level of the healthy women in pregnancy group was significantly higher than that in control group (2.50 vs 1.54 mU/L, P < 0.01). The positive rate of TPOAb in pregnancy women was lower than that in non-pregnancy women (3.3% vs 9.4%, P < 0.01). CONCLUSION: At the end of pregnancy, hypothyroidism accounts for most thyroid diseases. Thyroid autoimmunity is suppressed. PMID- 17083837 TI - [Influence of the depth of embryo transfer on pregnancy outcome in in vitro fertilization-embryo transfer]. AB - OBJECTIVE: To detect whether the depth of embryo transfer has influence on pregnancy outcome in in vitro fertilization-embryo transfer (IVF-ET). METHODS: The distance between the high echogenic transfer dot and the fundal endometrium was measured under guidance of transabdominal ultrasound. The average distance 0.75 cm was used to divide patients into two groups. Group 1 included 44 patients with a distance > 0.75 cm, while group 2 had 48 patients with a distance or =18 mm Hg and < or =40 mm Hg on maximum tolerated medical therapy. INTERVENTIONS: 350 mm(2) Baerveldt glaucoma implant or trabeculectomy with mitomycin C (MMC). MAIN OUTCOME MEASURES: IOP, visual acuity, and reoperation for glaucoma. RESULTS: A total of 212 eyes of 212 patients were enrolled, including 107 in the tube group and 105 in the trabeculectomy group. At one year, IOP (mean +/- SD) was 12.4 +/- 3.9 mm Hg in the tube group and 12.7 +/- 5.8 mm Hg in the trabeculectomy group (P = .73). The number of glaucoma medications (mean +/- SD) was 1.3 +/- 1.3 in the tube group and 0.5 +/- 0.9 in the trabeculectomy group (P < .001). The cumulative probability of failure during the first year of follow-up was 3.9% in the tube group and 13.5% in the trabeculectomy group (P = .017). CONCLUSIONS: Nonvalved tube shunt surgery was more likely to maintain IOP control and avoid persistent hypotony or reoperation for glaucoma than trabeculectomy with MMC during the first year of follow-up in the TVT Study. Both surgical procedures produced similar IOP reduction at one year, but there was less need for supplemental medical therapy following trabeculectomy with MMC. PMID- 17083911 TI - In-gel activity staining of oxidized nicotinamide adenine dinucleotide kinase by blue native polyacrylamide gel electrophoresis. AB - Oxidized nicotinamide adenine dinucleotide (NAD(+)) kinase (NADK, E.C. 2.7.1.23) plays an instrumental role in cellular metabolism. Here we report on a blue native polyacrylamide gel electrophoretic technique that allows the facile detection of this enzyme. The product, oxidized nicotinamide adenine dinucleotide phosphate (NADP(+)), formed following the reaction of NADK with NAD(+) and adenosine 5'-triphosphate was detected with the aid of glucose-6-phosphate dehydrogenase or NADP(+)-isocitrate dehydrogenase, iodonitrotetrazolium chloride, and phenazine methosulfate. The bands at the respective activity sites were excised and subjected to native and denaturing two-dimensional electrophoresis for the determination of protein levels. Hence this novel electrophoretic method allows the easy detection of NADK, a critical enzyme involved in pyridine homeostasis. Furthermore, this technique allowed the monitoring of the activity and expression of this kinase in various biological systems. PMID- 17083912 TI - A chemiluminescent metalloimmunoassay based on silver deposition on colloidal gold labels. AB - A sensitive chemiluminescent (CL) immunoassay of human immunoglobulin (IgG) which combined the inherent high sensitivity of CL analysis with the dramatic signal amplification of silver precipitation on colloidal gold tags was developed. First, the sandwich-type complex was formed in this protocol by the primary antibody immobilized on the polystyrene wells, the analyte in the sample, and the secondary antibody labeled with colloidal gold. Second, the colloidal gold was treated by an Ag(+) reduction solution, which resulted in the catalytic precipitation of silver on the surface of colloidal gold. Third, a large number of Ag(+) were oxidatively released in HNO(3) solution from the silver metal anchored on the sandwich-type complexes and then the human IgG was indirectly determined by a sensitive combined CL reaction of Ag(+)-K(2)S(2)O(8)-Mn(2+)- H(3)PO(4)-luminol. The chemiluminescence intensity depends linearly on the logarithm of the concentration of human IgG over the range of 0.02-50ngml(-1) and detection limit (3sigma) is 0.005ngml(-1) (i.e., approximately 3x10(-14)M, 3amol in 100-mul sample). This assay has been successfully applied to the determination of human IgG in human serum samples and showed great potential for numerous applications in immunoassay. PMID- 17083913 TI - Identification of a surface protein on human brain microvascular endothelial cells as vimentin interacting with Escherichia coli invasion protein IbeA. AB - Escherichia coli K1 is the most common gram-negative bacteria that cause meningitis during the neonatal period. The ibeA gene product in E. coli K1 has been characterized as a virulence factor that contributes to the binding to and invasion of brain microvascular endothelial cells (BMEC). Here, we identified a surface protein on human BMEC, vimentin, that interacts with the E. coli invasion protein IbeA. The binding sites of the IbeA-vimentin interaction are located in the 271-370 residue region of IbeA and the vimentin head domain. The regulatory protease factor Xa is able to cleave IbeA between R297 and K298 residues, and this cleavage abolishes the IbeA-vimentin interaction. PMID- 17083914 TI - Cytoprotective responses in the Mediterranean mussel exposed to Hg2+ and CH3Hg+. AB - This study aimed at elucidating whether organic and inorganic forms of mercury contamination differently affect HSP and MXR cytoprotective responses in Mytilus galloprovincialis. Changes of relative transcript levels were assessed in digestive glands of mussels exposed to 0.75 microM CH(3)Hg(+) or Hg(2+) for a maximum of 6 days. CH(3)Hg(+) inhibited MgHSP70 and induced MgHSC70 expression. As previously reported Hg(2+) stimulated MgHSP70 expression and induced MgHSC70 transcripts only after 6 days. The effect of CH(3)Hg(+) or Hg(2+) on the MXR system was investigated after partial sequencing of Pgp, Mrp2, and Mvp cDNAs. The multiple alignments showed a high sequence homology with the human and the available bivalve MXR-related proteins. All transcripts were up-regulated by CH(3)Hg(+), while transiently up-regulated by Hg(2+) after 8 h of exposure, and down-regulated thereafter. Present results suggest that the HSP70 and MXR systems may be simultaneously modulated by the same stress stimulus, and that different chemical forms of contamination evoke different cytoprotective responses. PMID- 17083915 TI - Inhibition of homologous recombination repair in irradiated tumor cells pretreated with Hsp90 inhibitor 17-allylamino-17-demethoxygeldanamycin. AB - In order to investigate the mechanism of radio-sensitization by an Hsp90 inhibitor 17-allylamino-17-demethoxygeldanamycin (17-AAG), we studied repair of DNA double strand breaks (DSBs) in irradiated human cells pre-treated with 17 AAG. DSBs are thought to be the critical target for radiation-induced cell death. Two human tumor cell lines DU145 and SQ-5 which showed clear radio-sensitization by 17-AAG revealed a significant inhibition of DSB repair, while normal human cells which did not show radio-sensitization by the drug indicated no change in the DSB repair kinetics with 17-AAG. We further demonstrated that BRCA2 was a novel client protein for Hsp90, and 17-AAG caused the degradation of BRCA2 and in turn altered the behavior of Rad51, a critical protein for homologous recombination (HR) pathway of DSB repair. Our data demonstrate for the first time that 17-AAG inhibits the HR repair process and could provide a new therapeutic strategy to selectively result in higher tumor cell killing. PMID- 17083916 TI - Critical role of the N-loop and beta1-strand hydrophobic clusters of RANTES derived peptides in anti-HIV activity. AB - HIV initiates its infectious cycle by docking to CD4 and a chemokine receptor, most commonly CCR5. RANTES, a natural CCR5 ligand, is a potent inhibitor of HIV 1. Despite the lack of structural information on the RANTES-CCR5 complex, determinants of HIV blockade were previously identified within the RANTES N-loop and beta1-strand regions. A prototype N-loop/beta1-strand peptide, named R11-29, contains two terminal hydrophobic stretches separated by a central hydrophilic region. Here, the role of the terminal hydrophobic clusters was investigated by means of amino acid substitutions or deletions. Most hydrophobic residues in these clusters were shown to be fundamental for the anti-HIV activity. However, increasing the hydrophobicity of the two clusters using non-natural amino acids did not significantly improve the potency of the peptides. These results may provide instrumental knowledge for the rational design of RANTES-derivative molecules with increased anti-HIV activity. PMID- 17083917 TI - Crystal structure of Bacillus subtilis S-adenosylmethionine:tRNA ribosyltransferase-isomerase. AB - The enzyme S-adenosylmethionine:tRNA ribosyltransferase-isomerase (QueA) is involved in the biosynthesis of the hypermodified tRNA nucleoside queuosine. It is unprecedented in nature as it uses the cofactor S-adenosylmethionine as the donor of a ribosyl group. We have determined the crystal structure of Bacillus subtilis QueA at a resolution of 2.9A. The structure reveals two domains representing a 6-stranded beta-barrel and an alpha beta alpha-sandwich, respectively. All amino acid residues invariant in the QueA enzymes of known sequence cluster at the interface of the two domains indicating the localization of the substrate binding region and active center. Comparison of the B. subtilis QueA structure with the structure of QueA from Thermotoga maritima suggests a high domain flexibility of this enzyme. PMID- 17083918 TI - Detection of cholesterol-rich microdomains in the inner leaflet of the plasma membrane. AB - The C-terminal domain (D4) of perfringolysin O binds selectively to cholesterol in cholesterol-rich microdomains. To address the issue of whether cholesterol rich microdomains exist in the inner leaflet of the plasma membrane, we expressed D4 as a fusion protein with EGFP in MEF cells. More than half of the EGFP-D4 expressed in stable cell clones was bound to membranes in raft fractions. Depletion of membrane cholesterol with beta-cyclodextrin reduced the amount of EGFP-D4 localized in raft fractions, confirming EGFP-D4 binding to cholesterol rich microdomains. Subfractionation of the raft fractions showed most of the EGFP D4 bound to the plasma membrane rather than to intracellular membranes. Taken together, these results strongly suggest the existence of cholesterol-rich microdomains in the inner leaflet of the plasma membrane. PMID- 17083919 TI - LKB1, an upstream AMPK kinase, regulates glucose and lipid metabolism in cultured liver and muscle cells. AB - LKB1 is a 50 kDa serine/threonine kinase that phosphorylates and activates the catalytic subunit of AMPK at its T-loop residue Thr 172. We prepared adenoviruses expressing the constitutive active (wild-type) form (CA) or dominant negative (kinase inactive, D194A mutant) form (DN) of LKB1 and overexpressed these proteins in cultured myotubes (C2C12 cells) and rat hepatoma cells (FAO cells). When analyzed by immunoblotting with the antibody against Thr172-phosphorylated AMPK, the phosphorylation of AMPK was increased (2.5-fold) and decreased (0.4 fold) in cells expressing CA and DN LKB1, respectively, as compared with Lac-Z expressing control cells. Immunoprecipitation experiments, using isoform-specific antibody, revealed these alterations of AMPK phosphorylation to be attributable to altered phosphorylation of AMPK alpha2, but not alpha1 catalytic subunits, strongly suggesting the alpha2 catalytic subunit to be the major substrate for LKB1 in mammalian cells. In addition, adiponectin or AICAR-stimulated AMPK phosphorylation was inhibited by overexpression of DN LKB1, while phenformin stimulated phosphorylation was unaffected. These results may explain the difference in AMPK activation mechanisms between AMP and phenformin, and also indicate that AMPK phosphorylation by LKB1 is involved in AMP-stimulated AMPK activation. As a downstream target for AMPK, AICAR-induced glucose uptake and ACCbeta phosphorylation were found to be significantly reduced in DN LKB1 expressing C2C12 cells. The expression of key enzymes for gluconeogenesis, glucose-6-phosphatase and phosphoenolpyruvate carboxykinase, was also dependent on LKB1 activities in FAO cells. These results demonstrate that LKB1 is a crucial regulator of AMPK activation in muscle and liver cells and, therefore, that LKB1 activity is potentially of importance to our understanding of glucose and lipid metabolism. PMID- 17083920 TI - 5HTTLPR polymorphism and enlargement of the pulvinar: unlocking the backdoor to the limbic system. AB - BACKGROUND: The 5HTTLPR genetic variant of the serotonin transporter (SERT), which consists of a long (SERT-l) and short (SERT-s) allele, has emerged as a major factor influencing emotional behavior and brain anatomy. The pulvinar nucleus of the thalamus projects to important limbic nuclei including the amygdala and cingulate cortex, is involved in the processing of stimuli with emotional content, and contains an abundance of SERT. METHODS: Stereological methods were used to measure pulvinar neuron number in postmortem tissue from major depressive disorder (n = 11), bipolar disorder (n = 11), schizophrenia (n = 12), and control (n = 15) specimens from the Stanley Foundation Neuropathology Consortium. The effect of SERT genotype on pulvinar volume and neuron number was investigated by using analysis of covariance. RESULTS: Analysis of covariance with diagnosis, SERT genotype, age, hemisphere, postmortem interval, and time-in formalin covariates identified a 20% increase in pulvinar neuron number and volume in SERT-ss subjects. CONCLUSIONS: The elevated number of pulvinar neurons in subjects with a SERT-ss genotype may serve to enhance subcortical input of emotionally relevant stimuli to the limbic system, providing a mechanism for the 5HTTLPR genetic variant to affect predisposition to conditions such as major depression. PMID- 17083921 TI - Ischemic acidosis causes apoptosis in coronary endothelial cells through activation of caspase-12. AB - OBJECTIVE: Myocardial ischemia has been shown to induce apoptosis of endothelial cells (EC). However, the mechanism of this endothelial injury is still poorly understood. To analyse the signaling pathway of ischemia-induced EC apoptosis was the aim of the present study. METHODS: The primary culture of rat coronary EC was exposed to simulated ischemia (glucose-free anoxia at pH(o) 6.4). Apoptosis was defined by staining of nuclei with Hoechst-33342 and TUNEL. Cytosolic Ca2+ and pH were measured with Fura-2 and BCECF, respectively. RESULTS: Apoptosis (29.2+/ 1.7% of cells) induced by exposure to simulated ischemia for 2 h was accompanied by cytosolic Ca2+ overload (1090+/-52 nmol/l) and acidosis (pHi = 6.52+/-0.13). Simulated ischemia had no significant effect on caspase-8 cleavage, but induced cleavage of caspase-3 and caspase-12 and led to a slight release of cytochrome C. Prevention of cytosolic acidosis (anoxia at pH(o) 7.4) had no effect on cytochrome C release, but significantly reduced apoptosis, attenuated cytosolic Ca2+ overload, and prevented cleavage of caspase-12. A similar effect was achieved by inhibition of Ca2+ release channels in the endoplasmic reticulum with ryanodine and xestospongin C. Knock-down of caspase-12 with small interfering RNA suppressed caspase-3 activation and reduced apoptotic cell number by about 70%. CONCLUSION: Acidosis, rather than anoxia, is an important trigger of apoptosis in EC under simulated ischemia. The main pathway of the simulated ischemia-induced apoptosis consists of the Ca2+ leak from the ER followed by activation of caspase 12 and caspase-3. PMID- 17083923 TI - Immunology's first priority dispute--an account of the 17th-century Rudbeck Bartholin feud. AB - Modern immunology has been notably free of public disputes over credit for major discoveries in this discipline. But the early recognition of the lymphatic system witnessed two examples of heated priority feuds. The first in the 17th-century concerned the greater anatomical organization of the system, while the second in the 18th-century concerned its function. This essay reviews the earlier of the two disputes, in which a Swedish medical student (Ole Rudbeck) charged a respected Danish Professor (T. Bartholin) with plagiarism and antedating his observations. Thus ethical issues in immunology predate modern times. How this discipline reached this point in its history is another focus of this essay and also an excuse to review briefly the anatomy of the lymphatic system. The influence of nationalistic pride on priority disputes is also discussed. PMID- 17083924 TI - Early specific free radical-related cytotoxicity of gas phase cigarette smoke and its paradoxical temporary inhibition by tar: An electron paramagnetic resonance study with the spin trap DEPMPO. AB - Electron paramagnetic resonance (EPR) spin trapping studies demonstrated aqueous tar particulate matter (TPM) and gas phase cigarette smoke (GPCS) to behave as different sources of free radicals in cigarette smoke (CS) but their cytotoxic implications have been only assessed in CS due to its relevance to the natural smoking process. Using a sensitive spin trapping detection with 5 (diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide (DEPMPO), this study compared the respective roles of CS- and GPCS-derived free radicals on smoke-induced cytotoxicity and lipid peroxidation of filtered and unfiltered, machine-smoked experimental and reference cigarettes yielding a wide range of TPM yields. In buffer bubbled with CS the DEPMPO/superoxide spin adduct was the major detected nitroxide. Use of appropriate control experiments with nitric oxide radical (NO*) or carbonyl sulfide, and a computer analysis of spin adduct diastereoisomery showed that the hydroxyl radical (HO*) adduct of DEPMPO seen in GPCS-bubbled was rather related to metal-catalyzed nucleophilic synthesis than to direct HO* trapping. Unexpectedly a protective effect of TPM on murine 3T3 fibroblasts was observed in early (<3h) free radical-, GPCS-induced cell death, and carbon filtering decreased free radical formation, toxicity and lipid peroxidation in three cell lines (including human epithelial lung cells) challenged with GPCS. These results highlight an acute, free radical-dependent, harmful mechanism specific to the GPCS phase, possibly involving NO* chemistry, whose physical or chemical control may be of great interest with the aim of reducing the toxicity of smoke. PMID- 17083922 TI - Defective T cell receptor-mediated signal transduction in memory CD4 T lymphocytes exposed to superantigen or anti-T cell receptor antibodies. AB - Lymphocytes must promote protective immune responses while still maintaining self tolerance. Stimulation through the T cell receptor (TCR) can lead to distinct responses in naive and memory CD4 T cells. Whereas peptide antigen stimulates both naive and memory T cells, soluble anti-CD3 antibodies and bacterial superantigens stimulate only naive T cells to proliferate and secrete cytokines. Further, superantigens, like staphylococcal enterotoxin B (SEB), cause memory T cells to become anergic while soluble anti-CD3 does not. In the present report, we show that signal transduction through the TCR is impaired in memory cells exposed to either anti-CD3 or SEB. A block in signaling leads to impaired activation of the kinase ZAP-70 so that downstream signals and cell proliferation do not occur. We further show that the signaling defect is unique to each agent. In anti-CD3-treated memory T cells, the src kinase Lck is only transiently activated and does not phosphorylate and activate ZAP-70. In SEB-treated memory T cells, ZAP-70 does not interact with the TCR/CD3 complex to become accessible to Lck. Finally, we provide evidence that alternative signaling pathways are initiated in SEB-treated memory cells. Altered signaling, indicated by an elevation in activity of the src kinase Fyn, may be responsible for memory cell anergy caused by SEB. Thus, differentiation of naive T cells into memory cells is accompanied by alterations in TCR-mediated signaling that can promote heightened recall immunity or specific tolerance. PMID- 17083925 TI - Effect of 17beta-estradiol on apoptosis, IGF system components and gelatinases A and B in prostate cancer cells (PC-3). AB - BACKGROUND: Previous studies have indicated that estrogen administration in the advanced stage of prostate cancer provide some benefits to the patients. Estrogen action was thought to be mediated via the blockade of the pituitary-testicular axis that effectively lowered the circulating levels of androgen and, thus, results in tumor regression; however, the effect of estrogens on prostate epithelial cells is still unclear. We investigated the effects of estradiol on insulin-like growth factor type I receptor (IGF-IR), IGF-binding protein 3 (IGFBP 3), IGFBP-4, and matrix metalloproteinase 2 (MMP-2) and MMP-9 in androgen independent prostate cancer cells (PC-3). METHODS: The cells were treated with different concentrations of estradiol (1, 10 and 100 nmol/l) for different time periods (24, 48, 72 and 96 h). Cell proliferation was assessed using MTT assay, and IGFBP-3 and IGFBP-4 were assessed using immunoradiometric and enzyme immunoassays, respectively. MMP-2, MMP-9 and IGF-IR expression levels were analyzed using western-blot analysis, and MMP-2 and MMP-9 activities were analyzed using gelatin zymography. Apoptosis was confirmed by Annexin V-FITC and acridine orange and ethidium bromide staining methods. DNA fragmentation studies were also performed. RESULTS: Cell proliferation assay revealed that 10 and 100 nmol/l estradiol concentrations inhibit the proliferation of PC-3 cells when incubated for 48-96 h. The secretory levels of IGFBP-3 and IGFBP-4 were increased significantly. The western-blot results showed that estradiol is capable of decreasing the expression of MMP-2 and MMP-9 significantly. Gelatin zymography showed that activities of MMP-2 and MMP-9 are decreased in estradiol-treated cells. Estradiol-induced apoptosis was studied using annexin V-binding and propidium iodide influx. Estradiol also induced nuclear fragmentation in higher doses (100 nmol/l) in PC-3 cells. CONCLUSION: Inhibition of MMPs in cancer cells and increased levels of IGFBP-3 and IGFBP-4 associated with apoptosis may be one of the targets for anticancer function of estradiol. Estradiol inhibits the proliferation of prostate cancer cells by inducing apoptosis. PMID- 17083926 TI - Comparison and the development of knowledge. AB - This paper considers the role of comparison in the development of knowledge. Results show that comparing similar objects makes them appear more similar. Comparing dissimilar objects, on the other hand does not make them appear more similar, and in some circumstances may make them appear less similar. The effect of comparison on similar items was especially striking since participants judged items to be more similar after comparison even if the comparison task was to list differences between the two items. Further, this effect appears specific to comparison and does not appear to be simply due to a "fleshing out" of object representations (listing properties of two objects without comparing the objects themselves served to increase the objects' similarity regardless of whether the objects were similar or dissimilar to start). This suggests that comparison may play a special role in partitioning bits of experience into categories, sharpening categorical boundaries, and otherwise helping us create conceptual structure above and beyond that offered by the world. PMID- 17083927 TI - Rapid learning of syllable classes from a perceptually continuous speech stream. AB - To learn a language, speakers must learn its words and rules from fluent speech; in particular, they must learn dependencies among linguistic classes. We show that when familiarized with a short artificial, subliminally bracketed stream, participants can learn relations about the structure of its words, which specify the classes of syllables occurring in first and last word positions. By studying the effect of familiarization length, we compared the general predictions of associative theories of learning and those of models postulating separate mechanisms for quickly extracting the word structure and for tracking the syllable distribution in the stream. As predicted by the dual-mechanism model, the preference for structurally correct items was negatively correlated with the familiarization length. This result is difficult to explain by purely associative schemes; an extensive set of neural network simulations confirmed this difficulty. Still, we show that powerful statistical computations operating on the stream are available to our participants, as they are sensitive to co occurrence statistics among non-adjacent syllables. We suggest that different learning mechanisms analyze speech on-line: A rapid mechanism extracting structural information about the stream, and a slower mechanism detecting statistical regularities among the items occurring in it. PMID- 17083928 TI - Modularity beyond perception: evidence from single task interference paradigms. AB - We combine the Dimension-Action (DA) model with translational models to account for both the Stroop and the flanker effects. The basic assumption of the model is that there are distinct visual modules, each of which is endowed with both perception and response selection processes. We contrast this model with an alternative widespread view, the standard view, according to which the same response selection processes are shared by all tasks. The two views have different predictions concerning the flanker and Stroop tasks. Seven experiments test these predictions. The first five experiments show that there is a fundamental difference between the typical Stroop and flanker effects. Moreover, these experiments show that words denoting colors can affect print colors only when they are required for naming or when participants verbally mediate the print color task. Experiments 6 and 7 show that an analogous interaction between color and shape exists in the flanker task. These experiments as well as previous studies are consistent with the DA model and the modular view and pose serious difficulties for the standard view. Wider implications of a visual modular architecture are discussed as well. PMID- 17083929 TI - Fibrosin, a novel fibrogenic cytokine, modulates expression of myofibroblasts. AB - Granulation tissue fibroblasts, or myofibroblasts are characterized by the presence of alpha smooth muscle actin fibers (alpha SMA). These specialized cells are involved in wound contraction and in retractile phenomena observed during fibrotic disease. Myofibroblasts have also been shown to play a role in embryonic development. Growth factors such as Transforming growth factor beta TGFbeta and Nerve growth factor (NGF) can modulate the differentiation of myofibroblasts. In this report, we show that in vitro application of fibrosin, a novel fibrogenic cytokine, stimulates expression of alpha SMA-producing cells at least four-fold above that observed in control cultures. In addition, administration of fibrosin in a wound healing model in mice stimulates increased numbers of myofibroblasts 7 days after injury, when compared with untreated, or, control, wounded mice. These results suggest that fibrosin plays an important role in up regulating the appearance of myofibroblasts during wound healing, and possibly in fibrotic diseases. It may, therefore, be important in the process of scarring. PMID- 17083930 TI - Molecular evidence and functional expression of multidrug resistance associated protein (MRP) in rabbit corneal epithelial cells. AB - Multidrug resistance associated protein (MRP) is a major family of efflux transporters involved in drug efflux leading to drug resistance. The objective of this study was to explore physical barriers for ocular drug absorption and to verify if the role of efflux transporters. MRP-2 is a major homologue of MRP family and found to express on the apical side of cell membrane. Cultured Rabbit Corneal Epithelial Cells (rCEC) were selected as an in vitro model for corneal epithelium. [14C]-erythromycin which is a proven substrate for MRP-2 was selected as a model drug for functional expression studies. MK-571, a known specific and potent inhibitor for MRP-2 was added to inhibit MRP mediated efflux. Membrane fraction of rCEC was used for western blot analysis. Polarized transport of [14C] erythromycin was observed in rCEC and transport from B-->A was significantly high than from A-->B. Permeability's increased significantly from A-->B in the presence of MK-571 and ketoconozole. Uptake of [14C]-erythromycin in the presence of MK-571 was significantly higher than control in rCEC. RT-PCR analysis indicated a unique and distinct band at approximately 498 bp corresponding to MRP 2 in rCEC and MDCK11-MRP-2 cells. Immunoprecipitation followed by Western Blot analysis indicated a specific band at approximately 190 kDa in membrane fraction of rCEC and MDCK11-MRP-2 cells. For the first time we have demonstrated high expression of MRP-2 in rabbit corneal epithelium and its functional activity causing drug efflux. RT-PCR, immunoprecipitation followed by Western blot analysis further confirms the result. PMID- 17083932 TI - Risk factors for age-related maculopathy are associated with a relative lack of macular pigment. AB - Macular pigment (MP) is composed of the two dietary carotenoids lutein (L) and zeaxanthin (Z), and is believed to protect against age-related maculopathy (ARM). This study was undertaken to investigate MP optical density with respect to risk factors for ARM, in 828 healthy subjects from an Irish population. MP optical density was measured psychophysically using heterochromatic flicker photometry, serum L and Z were quantified by HPLC, and dietary intake of L and Z was assessed using a validated food-frequency questionnaire. Clinical and personal details were also recorded, with particular attention directed towards risk factors for ARM. We report a statistically significant age-related decline in MP optical density (r2=0.082, p<0.01). Current and past smokers had lower average MP optical density than never smokers and this difference was statistically significant (p<0.01). Subjects with a confirmed family history of ARM had significantly lower levels of MP optical density than subjects with no known family history of disease (p<0.01). For each of these established risk factors, their statistically significant negative association with MP persisted after controlling for the other two, and also after controlling for other potentially confounding variables such as sex, cholesterol, dietary and serum L (p<0.01). In the absence of retinal pathology, and in advance of disease onset, the relative lack of MP seen in association with increasing age, tobacco use and family history of ARM supports the hypothesis that the enhanced risk that these variables represent for ARM may be attributable, at least in part, to a parallel deficiency of macular carotenoids. PMID- 17083931 TI - Preservation of photoreceptor morphology and function in P23H rats using an allele independent ribozyme. AB - To develop an allele independent ribozyme for the treatment of autosomal dominant retinitis pigmentosa (ADRP) associated with mutations in the rhodopsin (RHO) gene, a ribozyme targeting dog, mouse, human but not rat rhodopsin (RHO) mRNA was designed and tested in vitro. Activity of this ribozyme was tested in tissue culture by co-transfection of HEK 293 cells with plasmids expressing opsin mRNA and ribozyme, followed by quantitative RT-PCR to evaluate the level of RHO mRNA. For experiments in vivo, Rz525 driven by the mouse opsin proximal promoter was inserted in plasmids with AAV 2 terminal repeats (TR) and packaged in AAV serotype 5 capsids. AAV-Rz525 was injected subretinally into the right eyes of P23H rat pups. Left eyes were injected with virus expressing GFP from the identical promoter. Animals were analyzed at 4, 8 and 12 weeks post-injection by full field scotopic electroretinography (ERG). After 12 weeks, animals were sacrificed and retinas were dissected, fixed and sectioned. Rz525 had high catalytic activity in vitro and led to a 50% reduction of RHO mRNA in cells. AAV Rz525 injection into P23H transgenic rats led to significant preservation (about 50%) of scotopic ERG a- and b-wave amplitudes. Histological analysis showed an increased number of ONL nuclei in the central and superior retina of treated eyes relative to control eyes. RT-PCR analysis revealed 46% reduction of transgenic (mouse) RHO mRNA in right eyes relative to left eyes and no change in rat RHO mRNA. AAV5 delivery of Rz525 resulted in a partial rescue of the light response and structural preservation of photoreceptors in transgenic rats. This ribozyme may be a useful component of an RNA replacement gene therapy for ADRP. PMID- 17083933 TI - Cytochalasin D abolishes the schistosomicidal activity of praziquantel. AB - To test the hypothesis that calcium channels of schistosomes are the targets for the action of praziquantel, we subjected schistosomes in vitro to pharmacological agents capable of interfering with the functioning of calcium channels. After 1-h exposure to these agents, praziquantel was added and incubation continued overnight. Worms were then washed, resuspended in drug-free medium and observed during the following 7-10 days. About 50% of schistosomes pre-exposed to the calcium channel blockers nicardipine and nifedipine were able to survive a praziquantel concentration (3 microM) that normally killed the majority of adult male worms. Since the organization of the actin cytoskeleton controls the activity of calcium channels in a number of different systems, we also pre exposed schistosomes to the actin depolymerizing agent cytochalasin D. This treatment rendered the parasites completely refractory to the effects of very high praziquantel levels (up to 36 microM). These results are consistent with the hypothesis that schistosome calcium channels are involved in the mechanism of action of praziquantel. PMID- 17083934 TI - Leishmania (Kinetoplastida): species typing with isoenzyme and PCR-RFLP from cutaneous leishmaniasis patients in Ethiopia. AB - Cutaneous leishmaniasis (CL) is an increasing public health problem in Ethiopia. There is a concern that it is spreading with increased incidence. In this study, we used isoenzyme electrophoresis and internal transcribed spacer one (ITS1) PCR RFLP techniques to identify Leishmania species from CL patients in Ethiopia. We obtained isolates from 55 localized cutaneous leishmaniasis (LCL), 3 diffused cutaneous leishmaniasis (DCL) and 36 biopsy samples from 34 LCL and 2 DCL cases from All Africa Leprosy and Tuberculosis Rehabilitation and Training Center (ALERT) and clinically diagnosed CL cases from Ochollo village. Both isoenzyme and ITS1 PCR-RFLP techniques showed that Leishmania aethiopica (L. aethiopica) was the aetiologic agent in all cases. Our study also showed that ITS1 PCR-RFLP could identify Leishmania species from biopsy samples and suggests the method could be used for epidemiological surveillance of leishmaniasis in Ethiopia and for species-specific diagnosis. PMID- 17083935 TI - Plasmodium yoelii: the effect of second blood meal and anti-sporozoite antibodies on development and gene expression in the mosquito vector, Anopheles stephensi. AB - The sporogonic development of the malaria parasite takes place in the mosquito and a wide range of factors modulates it. Among those, the contents of the blood meal can influence the parasite development directly or indirectly through the mosquito response to the infection. We have studied the effect of a second blood meal in previously infected mosquitoes and the effect of anti-sporozoite immune serum on parasite development and mosquito response to the infection. The prevalence and intensity of infection and gene expression of both Plasmodium yoelii and Anopheles stephensi was analyzed. We verified that a second blood meal and its immune status interfere with parasite development and with Plasmodium and mosquito gene expression. PMID- 17083936 TI - Leishmania aethiopica: identification and characterization of cathepsin L-like cysteine protease genes. AB - There is limited information on the biology and pathogenesis of Leishmania aethiopica, causative agent of cutaneous leishmaniasis (CL) in Ethiopia. In this study we have identified and characterized two cathepsin L-like cysteine protease genes, Laecpa and Laecpb, from L. aethiopica. The predicted amino acid sequence of Laecpa and Laecpb is more than 75% identical with homologous cathepsin L-like cysteine protease genes of other Leishmania species and less than 50% identical with human cathepsin L. Laecpa is expressed predominantly in the stationary, and to a lower level, during the amastigote stage while Laecpb is specifically expressed in the stationary stage of L. aethiopica development. Phylogenetic analysis showed that the two genes are grouped into separate clades which are the result of gene duplication. The isolation of these genes will be useful in developing Leishmania species specific diagnostics for molecular epidemiological studies and serves as a first step to study the role of cysteine proteases in L. aethiopica pathogenesis. PMID- 17083937 TI - (2Alpha,3beta)-2,3-dihydroxyolean-12-en-28-oic acid, a new natural triterpene from Olea europea, induces caspase dependent apoptosis selectively in colon adenocarcinoma cells. AB - Triterpenoids are known to induce apoptosis and to be anti-tumoural. Maslinic acid, a pentacyclic triterpene, is present in high concentrations in olive pomace. This study examines the response of HT29 and Caco-2 colon-cancer cell lines to maslinic-acid treatment. At concentrations inhibiting cell growth by 50 80% (IC50HT29=61+/-1 microM, IC80HT29=76+/-1 microM and IC50Caco-2=85+/-5 microM, IC80Caco-2=116+/-5 microM), maslinic acid induced strong G0/G1 cell-cycle arrest and DNA fragmentation, and increased caspase-3 activity. However, maslinic acid did not alter the cell cycle or induce apoptosis in the non-tumoural intestine cell lines IEC-6 and IEC-18. Moreover, maslinic acid induced cell differentiation in colon adenocarcinoma cells. These findings support a role for maslinic acid as a tumour suppressant and as a possible new therapeutic tool for aberrant cell proliferation in the colon. In this report, we demonstrate for the first time that, in tumoural cancer cells, maslinic acid exerts a significant anti proliferation effect by inducing an apoptotic process characterized by caspase-3 activation by a p53-independent mechanism, which occurs via mitochondrial disturbances and cytochrome c release. PMID- 17083938 TI - Interaction of peroxidized cardiolipin with rat-heart mitochondrial membranes: induction of permeability transition and cytochrome c release. AB - Cardiolipin peroxidation plays a critical role in mitochondrial cytochrome c release and subsequent apoptotic process. Mitochondrial pore transition (MPT) is considered as an important step in this process. In this work, the effect of peroxidized cardiolipin on MPT induction and cytochrome c release in rat heart mitochondria was investigated. Treatment of mitochondria with micromolar concentrations of cardiolipin hydroperoxide (CLOOH) resulted in a dose-dependent matrix swelling, DeltaPsi collapse, release of preaccumulated Ca2+ and release of cytochrome c. All these events were inhibited by cyclosporin A and bongkrekic acid, indicating that peroxidized cardiolipin behaves as an inducer of MPT. Ca2+ accumulation by mitochondria was required for this effect. ANT (ADP/ATP translocator) appears to be involved in the CLOOH-dependent MPT induction, as suggested by the modulation by ligands and inhibitors of adenine nucleotide translocator (ANT). Together, these results indicate that peroxidized cardiolipin lowers the threshold of Ca2+ for MPT induction and cytochrome c release. This synergistic effect of Ca2+ and peroxidized cardiolipin on MPT induction and cytochrome c release in mitochondria, might be important in regulating the initial phase of apoptosis and also may have important implications in those physiopathological situations, characterized by both Ca2+ and peroxidized cardiolipin accumulation in mitochondria, such as aging, ischemia/reperfusion and other degenerative diseases. PMID- 17083939 TI - Reversible skeletal neuromuscular paralysis induced by different lysophospholipids. AB - Lysophosphatidylcholine rapidly paralyses the neuromuscular junction (NMJ), similarly to snake phospholipase A2 neurotoxins, implicating a lipid hemifusion pore transition in neuroexocytosis. The mode and kinetics of NMJ paralysis of different lysophospholipids (lysoPLs) in high or low [Mg2+] was investigated. The following order of potency was found: lysophosphatidylcholine>lysophosphatidylethanolamine>lysophosphatidic acid>lysophosphatidylserine>lysophosphatidylglycerol. The latter two lysoPLs closely mimic the profile of paralysis caused by the toxins in high [Mg2+]. This paralysis is fully reversed by albumin washing. These findings provide novel insights on the mode of action of snake neurotoxins and qualify lysoPLs as novel agents to study neuroexocytosis. PMID- 17083940 TI - Temporal expression of G-protein-coupled receptor 54 (GPR54), gonadotropin releasing hormones (GnRH), and dopamine receptor D2 (drd2) in pubertal female grey mullet, Mugil cephalus. AB - The G-protein-coupled receptor 54 (muGPR54) cDNA was cloned from the brain of the grey mullet, and its expression level, as well as those of the gonadotropin releasing hormones (GnRH1, GnRH2, GnRH3) and dopamine receptor D2 (drd2), in the brain, pituitary and ovary of pubertal fish (early, intermediate, advanced) were determined by real-time quantitative RT-PCR (QPCR). The muGPR54 cDNA has an open reading frame of 1140 bp with a predicted 380 amino acid peptide, containing seven putative transmembrane domains and putative N-glycosylation and protein kinase C phosphorylation sites. QPCR results showed that the early stage of puberty in grey mullet is characterized by significantly high levels of expression of GPR54, GnRH and drd2 in the brain relative to the intermediate and advanced stages, except for GnRH1 that increased at the advanced stage of puberty. In the pituitary, drd2 expression declined significantly at the advanced stage relative to levels at the intermediate stage. Ovarian expression of GPR54 significantly increased from the intermediate stage of puberty relative to the early stage while that of GnRH1 acutely increased at the advanced stage of puberty. The ovarian expression of drd2 decreased as puberty progressed, but the changes were not significant. The results suggest the possible role of GPR54 and GnRH in positively regulating pubertal development in grey mullet and the dopaminergic inhibition of reproductive function mediated by drd2. PMID- 17083941 TI - Resuscitative emergency thoracotomy in a Scandinavian trauma hospital--is it justified? AB - OBJECTIVE: Resuscitative emergency thoracotomy (ET) is of value in selected (penetrating) trauma patients. Current survival-estimates and recommended guidelines are based on data from the United States. However, reports from European trauma centres are lacking. We report the current experience from a Scandinavian trauma hospital. METHODS: Identification of all consecutive ETs performed during a 5-year period. Data on demographics, and injury severity score (ISS), mechanism and location were recorded. Physiological status on admission (revised trauma score, RTS) and probability of survival (Ps) were calculated. Signs of life (SOL) and need for closed-chest cardiopulmonary resuscitation (CC CPR) were recorded through the post-injury phase. RESULTS: Ten patients underwent ET with no survivors. The annual incidence of ET was 0.7 per 100,000 inhabitants during the study period, with an increasing trend during the last years (r=0.74, p=0.014). ETs were performed in 0.7% of all trauma admissions, and in 2.5% of all severely injured patients (ISS>or=16). Blunt mechanism dominated; only three had penetrating injuries. Most frequent location of major injury was "multiple" (n=4) and "thoracic" (n=4). The male to female ratio was 7:3. Median age was 51 years (range 21-77). Median ISS was 34.5 (range 26-75), indicating severely injured patients, with seriously deranged physiology (median RTS of 0.0, range 0-6.1) with poor chance of survival (median Ps of 4.4%, range 0-89.5%). Males had significantly lower RTS and Ps (p=0.007 and 0.03, respectively) than females. Eight patients had signs of life at some time post-injury, but only four in the emergency room. Six patients had both pre- and in-hospital CC-CPR. Four patients had additional surgery to ET. Two possible preventable deaths were identified (Ps) of 51 and 89%), one in a third trimester pregnancy. CONCLUSION: Emergency thoracotomy is a rarely performed procedure in a rather busy Scandinavian trauma hospital, and outcome is dismal. Reevaluation of our decision-making process concerning the use of emergency thoracotomy is needed. How survival data and clinical experience in Europe compare to current figures from North America deserves further attention. PMID- 17083942 TI - Demographic, social and economic variables that affect lower extremity injury outcomes. AB - While behavioral, demographic and vocational factors are commonly know as risk factors for sustaining a traumatic injury, less is known about the social, demographic and economic determinants of outcome following injury. The Lower Extremity Assessment Project (LEAP) identified a prospective cohort of 601 patients who were admitted to eight level I trauma centers in the United States for treatment of severe lower extremity trauma. Prospective data was accumulated on these individuals throughout their initial hospitalization and at regular follow-up intervals including a cohort at seven years post-injury. The results of the LEAP investigations showed that even at five to seven years following injury, reconstruction for the treatment of injuries to the lower extremity typically result in functional outcomes equivalent to those of the amputation. Clearly, factors other than the traditional variables such as fracture healing, joint function, and ability to ambulate have a profound effect on the patient's estimation of improvement. The purpose of this report is to summarize the data from the LEAP study and other investigations related to demographic, social and behavioral factors which impact outcome following lower extremity injury. PMID- 17083943 TI - Alarming rise in fall-induced severe head injuries among elderly people. AB - This study assessed the current trend in the number and incidence (per 100,000 persons) of fall-induced severe head injuries among the very old adults in Finland, an EU-country with a well-defined white population of 5.2 million, by taking into account all persons 80 years of age or older who were admitted to our hospitals for primary treatment of such injury in 1970-2004. The number of Finns aged 80 years or older with a fall-induced severe head injury increased considerably between the years 1970 and 2004, from 60 (women) and 25 (men) in 1970 to 745 (women), and 350 (men) in 2004. The relative increases were 1142 and 1300%, respectively. Across the study period, the age-adjusted incidence of injury also showed a clear increase from 1970 to 2004, from 168 to 506 in women (201% increase), and from 172 to 609 in men (254% increase). A similar finding was observed in age-specific incidences. If the age-adjusted incidence of injury continues to rise at the same rate as in 1970-2004 and the size of the 80 year old or older population of Finland increases as predicted (approximately 2.2-fold increase during the coming 25 years), the number of fall-induced severe head injuries in this population will be about 3.4-fold higher in the year 2030 than it was in 2004. In Finnish persons 80 years of age or older, the number of fall induced severe head injuries shows an alarming rise with a rate that cannot be explained merely by the demographic changes of the population. The finding underscores an increasing influence of falls on well-being of our elderly persons, and therefore, effective fall-prevention actions should be initiated to control this development. PMID- 17083944 TI - Chromatographic resolution of a salt into its parent acid and base constituents. AB - Based on the results of the earlier proposed process of separation of mixtures of mineral electrolytes by size-exclusion chromatography (SEC), it has been suggested that a mineral salt must spontaneously resolve, at least partially, into its parent acid and base constituents, provided that the separating media discriminates the anion and cation of the salt according to their size. Indeed, migration of a zone of an aqueous salt solution through a bed of neutral nanoporous hypercrosslinked polystyrene-type packing was shown to result in the generation of acidic and alkaline effluent fractions. The principle of spontaneous salt resolution has been extended to other types of discriminating interactions between the stationary phase and the two ions of the salt. The idea was exemplified by the resolution of ammonium acetate, due to hydrophobic retention of the acetate, into fractions enriched in ammoniac and then acetic acid. PMID- 17083945 TI - Diffusive sampling of methylene chloride with solid phase microextraction. AB - This study examined the characteristics of a solid phase microextraction (SPME) assembly as a passive sampler to determine the short-term exposure level (STEL) of methylene chloride. Two types of SPME fibers and six sampling-related factors were chosen and nested in an L(18) Taguchi's orthogonal array. Samples were thermally desorpted and analyzed by gas chromatograph equipped with an electron capture detector (GC/ECD). The use of 85-mum Carboxen/polydimethylsiloxane (Car/PDMS) fibers resulted in greater adsorbed mass, which was highly correlated with the product of concentration and sampling time (r>0.99, p<0.0001), than 85 microm polyacrylate fibers. The sampling rate (SR) of the 85-microm Carboxen/polydimethylsiloxane fibers was not significantly affected by variations in relative humidity (0-80%) and coexistent toluene (none to 100 ppm). Variance of sampling rate was predominantly attributed to the diffusive path length (86.4%) and sampling time (5.7%). With diffusive paths of 3, 10 and 15 mm, the sampling rates of 85-microm Carboxen/polydimethylsiloxane fibers for methylene chloride were 1.4 x 10(-2), 7.7 x 10(-3) and 5.1 x1 0(-3)mL min(-1), respectively. The measured sampling rates were greater than the theoretical values, and decreased with increment of sampling time until they came to constant. PMID- 17083946 TI - A study of retention and overloading of basic compounds with mixed-mode reversed phase/cation-exchange columns in high performance liquid chromatography. AB - The retention and overload of bases were studied on two new mixed-mode, silica based phases possessing ionic carboxylate functionalities of different acidity embedded within a hydrophobic ligand (SiELC Primesep). At low pH, good peak shapes were obtained for small solute mass, suggesting that the mere presence of a mixed-mode hydrophobic/ionic retention mechanism is not responsible for the poor peak shape that can occur on conventional reversed-phases with ionised silanols. Somewhat inferior, but still acceptable peak shape for bases was obtained on a column containing a mixture of discrete ion exchange and reversed phase particles (Hypersil Duet). In both types of column, the ionic sites favourably increased the capacity for ionised bases, reducing considerably the deterioration of peak shape with load observed with conventional RP columns. The combined ionic and reversed-phase interaction can give strong retention of bases under certain conditions, necessitating careful choice of stationary and mobile phase. PMID- 17083947 TI - Determination of aflatoxins in Chinese medicinal herbs by high-performance liquid chromatography using immunoaffinity column cleanup Improvement of recovery. AB - Although analytical methods are available for the determination of aflatoxins in medicinal herbs, none of them can be applied satisfactorily to all sample matrices. The difficulty arises from the complex chemical composition of the herbs. Recovery is generally low by using immunoaffinity column cleanup due to the acidity of the water extractive leading to a weakened binding affinity. As a solvent for dilution and neutralization, phosphate buffer saline is useful for certain herbs but not for others that have high acidity. The problem can be solved by using 0.1 M phosphate buffer, which has a higher buffering capacity and eliminates sodium chloride. The modified method was validated by the analysis of a certified reference material and shown to be useful for the determination of aflatoxins in herbal samples of high acidity. PMID- 17083948 TI - Determination of stoichiometric coefficients and apparent formation constants for beta-cyclodextrin complexes of trans-resveratrol using reversed-phase liquid chromatography. AB - The complexation of trans-resveratrol with beta-cyclodextrin (beta-CD) was investigated using reversed-phase liquid chromatography and mobile phases to which beta-CD was added. The decrease in the retention times with increasing concentrations of beta-CD (0-2.5 mM) showed that trans-resveratrol forms a 1:1 complex with beta-CD, while the apparent formation constants (K(F)) were strongly dependent of the water-methanol proportion of the mobile phase employed. A slight decrease in K(F) was observed at 50-55% water in the mobile phase but a large increase from 55 to 70% water. The values of K(F) for the trans-resveratrol-beta CD interaction decreased when the temperature was raised from 20 to 37 degrees C. In order to gain information about the mechanism aspect of the trans-resveratrol affinity for beta-CD, the thermodynamic parameters of the complexation were obtained. Complex formation of trans-resveratrol with beta-CD (DeltaG degrees = 17.01 kJ/mol) is largely driven by enthalpy (DeltaH degrees = -30.62 kJ/mol) and slightly entropy changes (DeltaS degrees = -45.68 J/mol K). PMID- 17083949 TI - Neuropharmacological profile of FrPbAII, purified from the venom of the social spider Parawixia bistriata (Araneae, Araneidae), in Wistar rats. AB - The aims of the present study were to investigate the anticonvulsant activity and behavioral toxicity of FrPbAII using freely moving Wistar rats. Moreover, the effectiveness of this compound against chemical convulsants was compared to that of the inhibitor of the GABAergic uptake, nipecotic acid. Our results show that FrPbAII was effective against seizures induced by the i.c.v. injection of pilocarpine (ED(50) = 0.05 microg/animal), picrotoxin (ED(50) = 0.02 microg/animal), kainic acid (ED(50) = 0.2 microg/animal) and the systemic administration of PTZ (ED(50) = 0.03 microg/animal). The anticonvulsant effect of FrPbAII differed from that of nipecotic acid in potency, as the doses needed to block the seizures were more than 10 folds lower. Toxicity assays revealed that in the rotarod, the toxic dose of the FrPbAII is 1.33 microg/animal, and the therapeutic indexes were calculated for each convulsant. Furthermore, the spontaneous locomotor activity of treated animals was not altered when compared to control animals but differed from the animals treated with nipecotic acid. Still, FrPbAII did not induce changes in any of the behavioral parameters analyzed. Finally, when tested for cognitive impairments in the Morris water maze, the i.c.v. injection of FrPbAII did not alter escape latencies of treated animals. These findings indicate that the novel GABA uptake inhibitor is a potent anticonvulsant with mild side-effects when administered to Wistar rats. PMID- 17083950 TI - Effects of anthropogenic seawater acidification on acid-base balance in the sea urchin Psammechinus miliaris. AB - The purple-tipped sea urchin, Psammechinus miliaris, was exposed to artificially acidified seawater treatments (pH(w) 6.16, 6.63 or 7.44) over a period of 8 days. Urchin mortality of 100% was observed at pH(w) 6.16 after 7 days and coincided with a pronounced hypercapnia in the coelomic fluid producing an irrecoverable acidosis. Coelomic fluid acid-base measures showed that an accumulation of CO(2) and a significant reduction in pH occurred in all treatments compared with controls. Bicarbonate buffering was employed in each case, reducing the resultant acidosis, but compensation was incomplete even under moderate environmental hypercapnia. Significant test dissolution was inferred from observable increases in the Mg(2+) concentration of the coelomic fluid under all pH treatments. We show that a chronic reduction of surface water pH to below 7.5 would be severely detrimental to the acid-base balance of this predominantly intertidal species; despite its ability to tolerate fluctuations in pCO(2) and pH in the rock pool environment. The absence of respiratory pigment (or any substantial protein in the coelomic fluid), a poor capacity for ionic regulation and dependency on a magnesium calcite test, make echinoids particularly vulnerable to anthropogenic acidification. Geological sequestration leaks may result in dramatic localised pH reductions, e.g. pH 5.8. P. miliaris is intolerant of pH 6.16 seawater and significant mortality is seen at pH 6.63. PMID- 17083951 TI - Nitrogen transport in the ectomycorrhiza association: the Hebeloma cylindrosporum Pinus pinaster model. AB - The function of the ectomycorrhizal mutualism depends on the ability of the fungal symbionts to take up nutrients (particularly nitrogen) available in inorganic and/or organic form in the soil and to translocate them (or their metabolites) to the symbiotic roots. A better understanding of the molecular mechanisms underlying nutrient exchanges between fungus and plant at the symbiotic interface is necessary to fully understand the function of the mycorrhizal symbioses. The present review reports the characterization of several genes putatively involved in nitrogen uptake and transfer in the Hebeloma cylindrosporum-Pinus pinaster ectomycorrhizal association. Study of this model system will further clarify the symbiotic nutrient exchange which plays a major role in plant nutrition as well as in resistance of plants against pathogens, heavy metals, drought stress, etc. Ultimately, ecological balance is maintained and/or improved with the help of symbiotic associations, and therefore, warrant further understanding. PMID- 17083952 TI - Cloning and characterization of chalcone synthase from the moss, Physcomitrella patens. AB - Since the early evolution of land plants from primitive green algae, flavonoids have played an important role as UV protective pigments in plants. Flavonoids occur in liverworts and mosses, and the first committed step in the flavonoid biosynthesis is catalyzed by chalcone synthase (CHS). Although higher plant CHSs have been extensively studied, little information is available on the enzymes from bryophytes. Here we report the cloning and characterization of CHS from the moss, Physcomitrella patens. Taking advantage of the available P. patens EST sequences, a CHS (PpCHS) was cloned from the gametophores of P. patens, and heterologously expressed in Escherichia coli. PpCHS exhibited similar kinetic properties and substrate preference profile to those of higher plant CHS. p Coumaroyl-CoA was the most preferred substrate, suggesting that PpCHS is a naringenin chalcone producing CHS. Consistent with the evolutionary position of the moss, phylogenetic analysis placed PpCHS at the base of the plant CHS clade, next to the microorganism CHS-like gene products. Therefore, PpCHS likely represents a modern day version of one of the oldest CHSs that appeared on earth. Further, sequence analysis of the P. patens EST and genome databases revealed the presence of a CHS multigene family in the moss as well as the 3'-end heterogeneity of a CHS gene. Of the 19 putative CHS genes, 10 genes are expressed and have corresponding ESTs in the databases. A possibility of the functional divergence of the multiple CHS genes in the moss is discussed. PMID- 17083953 TI - Hyperforin and its analogues inhibit CYP3A4 enzyme activity. AB - Literature indicates that herb-drug interaction of St. John's wort is largely due to increased metabolism of the co-administered drugs that are the substrates of cytochrome P450 (CYP) 3A4 enzyme, alteration of the activity and/or expression of the enzyme. The major St. John's wort constituents, acylphloroglucinols, were evaluated for their effects on CYP3A4 enzyme activity to investigate their roles in herb-drug interaction. Hyperforin and four oxidized analogues were isolated from the plant and fully characterized by mass spectral and NMR analysis. These acylphloroglucinols inhibited activity of CYP3A4 enzyme potently in the fluorometric assay using the recombinant enzyme. Furoadhyperforin (IC(50) 0.072 microM) was found to be the most potent inhibitor of CYP3A4 enzyme activity, followed by furohyperforin isomer 1 (IC(50) 0.079 microM), furohyperforin isomer 2 (IC(50) 0.23 microM), hyperforin (IC(50) 0.63 microM) and furohyperforin (IC(50) 1.3 microM). As the acylphloroglucinols are potent inhibitors of the CYP3A4 enzyme, their modulation of the enzyme activity is unlikely to be involved in increased drug metabolism by St. John's wort. PMID- 17083954 TI - Sperm morphology in five species of cicadettine cicadas (Hemiptera: Cicadomorpha: Cicadidae). AB - Mature spermatozoa from five species of cicadas of the subfamily Cicadettinae (Quintilia wealei, Melampsalta leucoptera, Stagira simplex, Xosopsaltria thunbergi and Monomatapa matoposa) were examined by light and electron microscopy. In each species sperm are elongate, aggregated into organized bundles with their heads embedded in a homogenous matrix to form spermatodesmata, and exhibit polymegaly. The head of the sperm consist of an anteriorly positioned conical acrosome that has a tubular substructure and a deep, posterior invagination that forms the subacrosomal space (eccentrically positioned anteriorly). The acrosome is flattened anteriorly; posteriorly it extends along either side of the nucleus as two tubular processes that gradually decrease in diameter. The filiform nucleus tapers anteriorly and intrudes into the subscrosomal space. Posteriorly the nucleus has a lateral invagination that houses material of the so-called centriolar adjunct. Posterior to the centriolar adjuct and the nucleus are two crystalline mitochondrial derivatives and a centriole, respectively, the latter giving rise to the axoneme, which has a 9 + 9 + 2 arrangement of microtubules. In these respects the sperm are similar to those of platypleurine cicadas. However, some features seem unique to cicadettines, including the structural organization of an enlarged centriolar adjunct and the dimensions of the tails. The enlarged centriolar adjunct has a lamella-like substructure and can be considered a synapomorphic character in the Cicadettinae. It is, therefore, potentially useful in the separation of this subfamily from the Cicadinae. In addition, the great length of the sperm nucleus of long-headed sperm in M. matoposa could be a synapomorphy of this genus and related taphurine and cicadettine species. PMID- 17083955 TI - Effect of proteasome inhibition on toxicity and CYP3A23 induction in cultured rat hepatocytes: comparison with arsenite. AB - Previous work in our laboratory has shown that acute exposure of primary rat hepatocyte cultures to non-toxic concentrations of arsenite causes major decreases in the DEX-mediated induction of CYP3A23 protein, with minor decreases in CYP3A23 mRNA. To elucidate the mechanism for these effects of arsenite, the effects of arsenite and proteasome inhibition, separately and in combination, on induction of CYP3A23 protein were compared. The proteasome inhibitor, MG132, inhibited proteasome activity, but also decreased CYP3A23 mRNA and protein. Lactacystin, another proteasome inhibitor, decreased CYP3A23 protein without affecting CYP3A23 mRNA at a concentration that effectively inhibited proteasome activity. This result, suggesting that the action of lactacystin is similar to arsenite and was post-transcriptional, was confirmed by the finding that lactacystin decreased association of DEX-induced CYP3A23 mRNA with polyribosomes. Both MG132 and lactacystin inhibited total protein synthesis, but did not affect MTT reduction. Arsenite had no effect on ubiquitination of proteins, nor did arsenite significantly affect proteasomal activity. These results suggest that arsenite and lactacystin act by similar mechanisms to inhibit translation of CYP3A23. PMID- 17083956 TI - Stimulus flicker alters interocular grouping during binocular rivalry. AB - When the two eyes are presented with sufficiently different stimuli, the stimuli will engage in binocular rivalry. During binocular rivalry, a subject's perceptual state alternates between awareness of the stimulus presented to the right eye and that presented to the left eye. There are instances in which competition is not eye-based, but instead takes place between stimulus features, as is the case in flicker and switch rivalry (F&S). Here we investigate another such instance, interocular grouping, using a Diaz-Caneja type stimulus in conjunction with synchronous stimulus flicker. Our results indicate that stimulus flicker increases the total duration of interocularly bound percepts, and that this effect occurs for a range of temporal flicker frequencies. Furthermore, the use of contrast-inversion flicker causes a decrease of total dominance duration of the interocularly bound percepts. We argue that different flickering regimes can be used to differentially stimulate lower and higher levels of visual processing involved in binocular rivalry. We propose that the amount of interocularly combined pattern-completed percept can be regarded as a measure of the level at which binocular rivalry is resolved. PMID- 17083957 TI - The pursuit theory of motion parallax. AB - Although motion parallax is closely associated with observer head movement, the underlying neural mechanism appears to rely on a pursuit-like eye movement signal to disambiguate perceived depth sign from the ambiguous retinal motion information [Naji, J. J., & Freeman, T. C. A. (2004). Perceiving depth order during pursuit eye movement. Vision Research, 44, 3025-3034; Nawrot, M. (2003). Eye movements provide the extra-retinal signal required for the perception of depth from motion parallax. Vision Research, 43, 1553-1562]. Here, we outline the evidence for a pursuit signal in motion parallax and propose a simple neural network model for how the pursuit theory of motion parallax might function within the visual system. The first experiment demonstrates the crucial role that an extra-retinal pursuit signal plays in the unambiguous perception of depth from motion parallax. The second experiment demonstrates that identical head movements can generate opposite depth percepts, and even ambiguous percepts, when the pursuit signal is altered. The pursuit theory of motion parallax provides a parsimonious explanation for all of these observations. PMID- 17083958 TI - A single motion system suffices for global-motion perception. AB - Global-motion perception is the perception of coherent motion in a noisy motion stimulus. Thresholds for coherent motion perception were measured for different combinations of signal and noise speeds. Previous research [Edwards, M., Badcock, D. R., & Smith, A. T. (1998). Independent speed-tuned global-motion systems. Vision Research, 38 (11), 1573-1580; Khuu, S. K., & Badcock, D. R. (2002). Global speed processing: evidence for local averaging within, but not across two speed ranges. Vision Research, 42 (28), 3031-3042.] showed that thresholds were elevated when signal and noise speeds were similar, but not when they were different. The regions of increased threshold values for low and high signal speeds showed little overlap. On the basis of this evidence two independent speed tuned systems were proposed: one for slow and one for fast-motion. However, in those studies only two signal speeds were used. We expanded the results by measuring threshold-curves for four different signal speeds. Considerable overlap of the threshold-curves was found between conditions. These results speak against a bipartite global-motion system. Model simulations indicate that present and previous experimental results can be produced by a single motion system providing that the mechanisms within it are speed-tuned. PMID- 17083959 TI - Direct crystallographic observation of an acyl-enzyme intermediate in the elastase-catalyzed hydrolysis of a peptidyl ester substrate: Exploiting the "glass transition" in protein dynamics. AB - The crystal structure of the acyl complex of porcine pancreatic elastase with its peptidyl ester substrate N-acetyl-ala-ala-ala-methyl ester (Ac(Ala)3OMe) has been determined at 2.5 A resolution. The complex was stabilized by exploiting the "glass transition" in protein dynamics that occurs at around -53 degrees C (220 K). Substrate was flowed into the crystal in a cryoprotective solvent above this temperature, and then the crystal was rapidly cooled to a temperature below the transition to trap the species that formed. The use of a flow cell makes the experiment a kinetic one and means that the species prior to the rate determining transition state has a chance to accumulate. The resulting crystal structure shows an acyl-enzyme intermediate in which the leaving group is absent and the carbonyl carbon of the C-terminal alanine residue is covalently bound to the gamma oxygen of the active site serine. The ester carbonyl shows no significant distortion from planarity, with the carbonyl oxygen forming one hydrogen bond with the oxyanion hole. The tripeptide is bound in an extended antiparallel beta sheet with main chain residues of the enzyme. The geometry and interactions of this acyl-enzyme suggest that it represents a productive intermediate. To test this hypothesis, the same crystal was then warmed above the glass transition temperature and a second data set was collected. The resulting electron density map shows no sign of the substrate, indicating hydrolysis of the intermediate followed by product release. This experiment provides direct evidence for the importance of dynamic properties in catalysis and also provides a blueprint for the stabilization of other short-lived species for direct crystallographic observation. PMID- 17083960 TI - Structural and thermodynamic studies of simple aldose reductase-inhibitor complexes. AB - The competitive inhibition constants of series of inhibitors related to phenylacetic acid against both wild-type and the doubly mutanted C298A/W219Y aldose reductase have been measured. Van't Hoff analysis shows that these acids bind with an enthalpy near -6.8 kcal/mol derived from the electrostatic interactions, while the 100-fold differences in binding affinity appear to be largely due to entropic factors that result from differences in conformational freedom in the unbound state. These temperature studies also point out the difference between substrate and inhibitor binding. X-ray crystallographic analysis of a few of these inhibitor complexes both confirms the importance of a previously described anion binding site and reveals the hydrophobic nature of the primary binding site and its general plasticity. Based on these results, N glycylthiosuccinimides were synthesized to demonstrate their potential in studies that probe distal binding sites. Reduced alpha-lipoic acid, an anti-oxidant and therapeutic for diabetic complications, was shown to bind aldose reductase with a binding constant of 1 microM. PMID- 17083961 TI - Synthesis with good enantiomeric excess of both enantiomers of alpha-ketols and acetolactates by two thiamin diphosphate-dependent decarboxylases. AB - In addition to the decarboxylation of 2-oxo acids, thiamin diphosphate (ThDP) dependent decarboxylases/dehydrogenases can also carry out so-called carboligation reactions, where the central ThDP-bound enamine intermediate reacts with electrophilic substrates. For example, the enzyme yeast pyruvate decarboxylase (YPDC, from Saccharomyces cerevisiae) or the E1 subunit of the Escherichia coli pyruvate dehydrogenase complex (PDHc-E1) can produce acetoin and acetolactate, resulting from the reaction of the central thiamin diphosphate bound enamine with acetaldehyde and pyruvate, respectively. Earlier, we had shown that some active center variants indeed prefer such a carboligase pathway to the usual one [Sergienko, Jordan, Biochemistry 40 (2001) 7369-7381; Nemeria et al., J. Biol. Chem. 280 (2005) 21,473-21,482]. Herein is reported detailed analysis of the stereoselectivity for forming the carboligase products acetoin, acetolactate, and phenylacetylcarbinol by the E477Q and D28A YPDC, and the E636A and E636Q PDHc E1 active-center variants. Both pyruvate and beta-hydroxypyruvate were used as substrates and the enantiomeric excess was analyzed by a combination of NMR, circular dichroism and chiral-column gas chromatographic methods. Remarkably, the two enzymes produced a high enantiomeric excess of the opposite enantiomer of both acetoin-derived and acetolactate-derived products, strongly suggesting that the facial selectivity for the electrophile in the carboligation is different in the two enzymes. The different stereoselectivities exhibited by the two enzymes could be utilized in the chiral synthesis of important intermediates. PMID- 17083963 TI - Adsorption and oxidation of fluoroquinolone antibacterial agents and structurally related amines with goethite. AB - Seven members (ciprofloxacin, enrofloxacin, norfloxacin, ofloxacin, lomefloxacin, pipemidic acid, and flumequine) of the popular fluoroquinolone antibacterial agents (FQs) were found to adsorb strongly to goethite with 50-76% of the added FQ adsorbed under the experimental conditions. The adsorption isotherms fitted well to the Langmuir model. Adsorption was accompanied by slow oxidation of the FQs (except for flumequine) by goethite yielding a range of hydroxylated and dealkylated products. The oxidation kinetics showed different stages in reaction rate, mostly likely caused by accumulation of Fe(II) species on the oxide surface that slowed the reaction. Structurally related amines 1-phenylpiperazine, N phenylmorpholine, aniline, and N,N-dimethylaniline were found to be oxidized by goethite without significant adsorption. The results strongly indicate that the carboxylic group of FQs is critical for adsorption while the piperazine ring is susceptible to oxidation. A radical mechanism is proposed for the oxidation of FQs by goethite which involves formation of a surface complex between the FQ and surface-bound Fe(III) through adsorption, and initial oxidation at the piperazinyl N1 atom to form radical intermediates that ultimately lead to the final products. This study indicates that Fe oxides in aquatic sediments may well play an important role in the natural attenuation of fluoroquinolone antibacterial agents. PMID- 17083962 TI - Fate of sex hormones in two pilot-scale municipal wastewater treatment plants: conventional treatment. AB - The fate of seven sex hormones (estrone (E1), estradiol (E2), estriol (E3), ethinylestradiol (EE2), testosterone, androstenedione, and progesterone) was determined in two pilot-scale wastewater treatment plants operated under conventional loading conditions. The levels of hormones in both the liquid and the solid matrixes of the plants were determined. Each of the two 20-l/h pilot scale plants consisted of a primary clarifier followed by a three-stage aeration tank and a final clarifier. The primary sludge and the waste activated sludge (WAS) were digested anaerobically in one pilot plant and aerobically in the other. The pilot plants were fed a complex synthetic wastewater spiked with the hormones. Levels of testosterone, androstenedione and progesterone were close to method detection limit (MDL) concentrations in the final and digester effluents (both liquid and solid phases) and were considered as completely removed. Average mass flux removals from the liquid streams (plant influent minus secondary clarifier effluent) for the natural estrogens were 82% for E1, 99% for E2, and 89% for (E1+E2). An average overall removal of only 42% was achieved for EE2. These values reflect removals averaged for the two pilot plants. PMID- 17083964 TI - Effect of hydrocarbon pollution on the microbial properties of a sandy and a clay soil. AB - The aim of this work was to ascertain the effects of different types of hydrocarbon pollution on soil microbial properties and the influence of a soil's characteristics on these effects. For this, toxicity bioassays and microbiological and biochemical parameters were studied in two soils (one sandy and one clayey) contaminated at a loading rate of 5% and 10% with three types of hydrocarbon (diesel oil, gasoline and crude petroleum) differing in their volatilisation potential and toxic substance content. Soils were maintained under controlled conditions (50-70% water holding capacity, and room temperature) for six months and several microbiological and toxicity parameters were monitored 1, 60, 120 and 180 days after contamination. The toxic effects of hydrocarbon contamination were greater in the sandy soil. Hydrocarbons inhibited microbial biomass, the greatest negative effect being observed in the gasoline-polluted sandy soil. In both soils crude petroleum and diesel oil contamination increased microbial respiration, while gasoline had little effect on this parameter, especially in the sandy soil. In general, gasoline had the highest inhibitory effect on the hydrolase activities involved in N, P or C cycles in both soils. All contaminants inhibited hydrolase activities in the sandy soil, while in the clayey soil diesel oil stimulated enzyme activity, particularly at the higher concentration. In both soils, a phytotoxic effect on barley and ryegrass seed germination was observed in the contaminated soils, particularly in those contaminated with diesel or petroleum. PMID- 17083965 TI - Microbial degradation and impact of Bracken toxin ptaquiloside on microbial communities in soil. AB - The carcinogenic and toxic ptaquiloside (PTA) is a major secondary metabolite in Bracken fern (Pteridium aquilinum (L.) Kuhn) and was hypothesized to influence microbial communities in soil below Bracken stands. Soil and Bracken tissue were sampled at field sites in Denmark (DK) and New Zealand (NZ). PTA contents of 2.1 +/- 0.5 mg g(-1) and 37.0 +/- 8.7 mg g(-1) tissue were measured in Bracken fronds from DK and NZ, respectively. In the two soils the PTA levels were similar (0-5 microg g(-1) soil); a decrease with depth could be discerned in the deeper B and C horizons of the DK soil (weak acid sandy Spodosol), but not in the NZ soil (weak acid loamy Entisol). In the DK soil PTA turnover was predominantly due to microbial degradation (biodegradation); chemical hydrolysis was occurring mainly in the uppermost A horizon where pH was very low (3.4). Microbial activity (basal respiration) and growth ([3H]leucine incorporation assay) increased after PTA exposure, indicating that the Bracken toxin served as a C substrate for the organotrophic microorganisms. On the other hand, there was no apparent impact of PTA on community size as measured by substrate-induced respiration or composition as indicated by community-level physiological profiles. Our results demonstrate that PTA stimulates microbial activity and that microorganisms play a predominant role for rapid PTA degradation in Bracken-impacted soils. PMID- 17083966 TI - Development and validation of a new fluorescence-based bioassay for aquatic macrophyte species. AB - Bioassays with unicellular algae are frequently used as ecotoxicological test systems to evaluate the toxicity of contaminated environmental samples or chemicals. In contrast, aquatic macrophyte test systems are still rarely used as they are laborious to handle because species exhibit distinct ecological requirements. The aim of this study was to establish a fast and reproducible measuring system for aquatic macrophyte species to overcome those limitations for use. Thus, a newly developed pulse-amplitude modulated chlorophyll fluorometer (Imaging-PAM) was applied as an effect detection in short-term bioassays with aquatic macrophyte species. This multiwell-plate-based measuring device enables the incubation and measurement of up to 24 samples in parallel. The Imaging-PAM was used (i) to establish and validate the sensitivity of the test systems to three Photosystem II (PSII) inhibitors (atrazine, prometryn, isoproturon), (ii) to compare the test systems with established biotests for macrophytes and (iii) to define necessary time scales in aquatic macrophyte testing. The results showed that fluorescence-based measurements with the Imaging-PAM allow rapid and parallel analysis of large amounts of aquatic macrophyte samples and of toxicants effects of the PSII inhibitors tested on aquatic macrophytes. Measurements revealed a good correlation between obtained median effective concentrations (EC50s) for the new and the established biotest systems. Hence, the Imaging-PAM measuring device is a promising tool to allow fast chemical effect screening for high amounts of samples with little time and material and thus offers scope for high-throughput biotesting using aquatic macrophyte species. PMID- 17083967 TI - Both trivalent and hexavalent chromium strongly alter in vitro germination and ultrastructure of kiwifruit pollen. AB - Due to its widespread industrial use, chromium is considered a dangerous environmental pollutant. It is known to inhibit plant growth and development. The present study provides the first evidence of the toxicity of this metal on the male haploid generation of a higher plant. Both Cr(III) and Cr(VI) species, supplied as CrCl(3) and CrO(3), respectively, exerted a strong dose-dependent inhibitory effect on kiwifruit pollen tube emergence and growth. Cr(III) resulted more effective than Cr(VI) in the 16-75microM interval; moreover, complete inhibition of germination was attained at much lower doses than Cr(VI). Also tube morphology was affected. While the plasma membrane was still undamaged in the large majority of the treated pollen grains, dramatic ultrastructural alterations were induced by chromium including chromatin condensation, swelling of mitochondria, cytoplasmic vacuolization, and perturbed arrangement of endoplasmic reticulum cisternae. Thus, it seems that the impact of the two chromium species on kiwifruit pollen may result in severe compromission of both essential structures and functions of the male gametophyte. PMID- 17083968 TI - The orang-utan mating system and the unflanged male: A product of increased food stress during the late Miocene and Pliocene? AB - The orang-utan is unique among apes in having an unusually long male developmental period and two distinct adult male morphs (flanged and unflanged), which generally, but not exclusively, employ different reproductive strategies (call-and-wait vs. sneak-and-rape). Both morphs have recently been shown to have roughly similar levels of reproductive success in the one site where such a study has been conducted. This is in stark contrast to the unimale polygynous gorilla, in which dominant males sire almost all infants. Despite this, evidence on sexual dimorphism, life history, diet, and socioecology of extant and extinct apes, as well as the ontogeny, reproductive morphology, and physiology of extant apes, all indicate that the orang-utan's present-day mating system most likely evolved from a gorilla-like base, with one dominant male guarding a harem of females. The available evidence indicates that, due chiefly to the likely onset of the El Nino Southern Oscillation (generally regarded as the trigger for mast fruiting in dipterocarps) approximately 3-5Ma, southeast Asian forests would have begun to experience longer and more severe periods of low food availability. This change in food availability would have meant that full-time gregariousness was no longer energetically tolerable and, as a result, females dispersed more widely in search of food and adult/flanged males were no longer able to effectively guard a harem of females. A niche for a quiet, quick, opportunistic "sexual predator" (i.e., the unflanged male) then became available. This finding implies that, despite being anatomically quite chimpanzee-like, the ancestral hominoid probably had a social and mating system more similar to the gorilla than any other living ape. PMID- 17083969 TI - Metal-binding proteins and peptides in the aquatic fungi Fontanospora fusiramosa and Flagellospora curta exposed to severe metal stress. AB - The production of thiol-containing proteins/peptides and its role in metal binding was examined in the aquatic hyphomycetes Fontanospora fusiramosa and Flagellospora curta exposed to Cu, Cd, or Zn at concentrations inhibiting the biomass production in 80%. Heat-treated cell-free extracts were separated by size exclusion chromatography and the thiol and metal content in the fractions was determined. F. curta, the species tolerant to metals, showed higher absolute levels of thiol compounds, which bound higher amounts of Cu and Cd than F. fusiramosa. Peptides with very low molecular weight (<9 kDa), most likely glutathione and phytochelatins, were the major Cu- and Zn-binding components in both species of aquatic hyphomycetes. In most cases, proteins with high molecular weight (>26 kDa) were induced by metal ions and they were the major Cd-binding component in both species. Proteins with characteristics of metallothioneins were also induced by exposure to metals in both species, but they showed a minor role in metal-binding, suggesting they might have other functions in fungal cells. PMID- 17083970 TI - Uterine sarcoma occurring in a premenopausal patient after uterine artery embolization: a case report and review of the literature. AB - OBJECTIVE: We report a case of uterine leiomyosarcoma occurring after uterine artery embolization and review the previously published cases. We estimate the incidence of sarcoma after UAE, the mean diagnostic delay in young women undergoing this procedure and review the potential and limits of preoperative procedures in diagnosing uterine sarcomas. CASE REPORT: A 35-year-old woman had an early failure after UAE. She underwent surgery 13 months after the procedure. Final pathologic report was consistent with uterine leiomyosarcoma. CONCLUSION: Incidence of uterine sarcomas after UAE is low, probably similar to that of misdiagnosed leiomyosarcomas in women undergoing surgery for presumed symptomatic leiomyomas. Therefore a relation between the procedure and the malignancy seems to be very unlikely. Diagnostic delay in menstruated women younger than 50 undergoing UAE for presumed symptomatic leiomyoma ranges between 13 and 15 months. The safest procedure for women who fail the conservative management of leiomyoma with UAE is surgical, allowing for definitive pathologic exclusion of malignancy. PMID- 17083972 TI - Selective alexia and agraphia sparing numbers-a case study. AB - We report a patient (MT) with a highly specific alexia affecting the identification of letters and words but not numbers. He shows a corresponding deficit in writing: his letter writing is impaired while number writing and written calculation is spared. He has no aphasia, no visuo-perceptual or constructional difficulties, or other cognitive deficits. A similar pattern of performance has to our knowledge only been reported once before [Anderson, S. W., Damasio, A. R., & Damasio, H. (1990). Troubled letters but not numbers. Domain specific cognitive impairments following focal damage in frontal cortex. Brain, 113, 749-766]. This study shows that letter and number reading are dependent on dissociable processes. More interestingly, it points to a common mechanism subserving the perception and production of letters. We suggest that a deficit in a visuo-motor network containing knowledge of the physical shape of letters might explain the pattern of performance displayed by MT. PMID- 17083973 TI - Cancer testis/germline antigens (CT/CG-antigens) in leukemias. PMID- 17083975 TI - CD19-/CD3-bispecific antibody of the BiTE class is far superior to tandem diabody with respect to redirected tumor cell lysis. AB - Many kinds of bispecific antibodies recruiting T cells for cancer therapy have been developed. Side-by-side comparison has shown that CD19-/CD3-bispecific antibodies of the diabody, tandem diabody (Tandab) and quadroma format had similar cytotoxic activity, with Tandab being the most active format. Tandab has also been claimed to be superior to single-chain (sc) Fv-based bispecific constructs although data from a side-by-side comparison are not available. In this study, we compared side-by-side MT103 (bscCD19xCD3), a single-chain bispecific antibody of the BiTE class, with a CD19-/CD3-bispecific representative of the Tandab class. Based on literature data, we have constructed, produced and characterized the LL linker version of Tandab, which was reported to be the most active version of Tandab proteins. A dimeric protein of 114kDa was obtained that showed proper bispecific binding to CD3- and CD19-positive cells and could redirect both pre-stimulated and unstimulated human T cells for lysis of human B lymphoma lines Raji, MEC-1 and Nalm-6. Raji cells were lysed at a half-maximal concentration (EC50) of 10 nM Tandab using pre-stimulated T cells, which closely matched the published activity of LL-Tandab with this particular cell line. MT103 had between 700- and 8000-fold higher efficacy than Tandab for redirected lysis of the three human B lymphoma lines. These data demonstrate that under identical experimental conditions, the BiTE format has far superior activity compared to the Tandab format and is also superior to conventional diabody and quadroma formats. The extraordinary potency of the BiTE class and its representative MT103 may translate into improved anti-tumor activity, lower dosing and lower costs of production compared to other bispecific antibody formats. PMID- 17083974 TI - A qualitative analysis of the use of community treatment orders in Saskatchewan. AB - This study examined the opinions of patients who have been placed on a community treatment order (CTO), their relatives, mental health clinicians and representatives of community agencies about the use of CTOs in Saskatchewan. Patients were assessed using indepth interviews, while their relatives, mental health professionals and representatives of community agencies took part in facilitated focus groups. Patients had contradictory feelings about CTOs. Most experienced some degree of coercion while on the orders but many believed that CTOs provided necessary structure in their lives. Clinicians were more consistently positive but recognized the difficult choices in balancing the subject's right to self-determination with the benefits of a treatment order. Family members viewed CTOs as necessary to control a chaotic situation caused by the subject's limited insight. PMID- 17083976 TI - DNA-binding and molecular mechanics modelling studies of the bulky chiral platinum(II) complex [PtCl(2)(mepyrr)] (mepyrr=N-methyl-2 aminomethylpyrrolidine). AB - Detailed studies were carried out on the binding of the enantiomers of [PtCl(2)(mepyrr)] (mepyrr=N-methyl-2-aminomethylpyrrolidine) to dG, d(GpG) and a 52-mer oligonucleotide. The pyrrolidine ligand structure was found to be neither sufficiently rigid nor bulky to enforce a single chirality at the exocyclic amine site in this complex, resulting in the presence of diastereomers that complicated the binding studies. Reaction of the (GpG) dinucleotide with R- and S [PtCl(2)(mepyrr)] resulted in formation of four [Pt{d(GpG)}(mepyrr)] isomers for each enantiomer as a consequence of the existence of two orientational isomers and two diastereomers. These isomers formed in different amounts most likely as a consequence of the unequal formation of the diastereomers together with stereoselectivity induced by interactions between the dinucleotide and the mepyrr ligand. The [PtCl(2)(mepyrr)] complexes displayed stereoselectivity and enantioselectivity in their reactions with a 52-mer duplex designed to allow formation of only GpG intrastrand adducts. All four bifunctional adducts formed for each enantiomer, providing further evidence of the lack of directing ability of the ligand in formation of the 1,2-intrastrand adduct. Significant amounts of monofunctional species remained in these assays suggesting that the introduction of the methyl substituent to the exocyclic amine inhibited ring-closure to the bifunctional adduct. This was not sufficient to achieve enantiospecificity, but in the case of the R-enantiomer, one of the bifunctional adducts formed in only small amounts. PMID- 17083977 TI - Spectroscopic investigations of intermediates in the reaction of cytochrome P450(BM3)-F87G with surrogate oxygen atom donors. AB - Rapid mixing of substrate-free ferric cytochrome P450(BM3)-F87G with m chloroperoxybenzoic acid (mCPBA) resulted in the sequential formation of two high valent intermediates. The first was spectrally similar to compound I species reported previously for P450(CAM) and CYP 119 using mCPBA as an oxidant, and it featured a low intensity Soret absorption band characterized by shoulder at 370nm. This is the first direct observation of a P450 compound I intermediate in a type II P450 enzyme. The second intermediate, which was much more stable at pH values below 7.0, was characterized by an intense Soret absorption peak at 406nm, similar to that seen with P450(CAM) [T. Spolitak, J.H. Dawson, D.P. Ballou, J. Biol. Chem. 280 (2005) 20300-20309]. Double mixing experiments in which NADPH was added to the transient 406nm-absorbing intermediate resulted in rapid regeneration of the resting ferric state, with the flavins of the flavoprotein domain in their reduced state. EPR results were consistent with this stable intermediate species being a cytochrome c peroxidase compound ES-like species containing a protein-based radical, likely localized on a nearby Trp or Tyr residue in the active site. Iodosobenzene, peracetic acid, and sodium m-periodate also generated the intermediate at 406nm, but not the 370nm intermediate, indicating a probable kinetic barrier to accumulating compound I in reactions with these oxidants. The P450 ES intermediate has not been previously reported using iodosobenzene or m-periodate as the oxygen donor. PMID- 17083971 TI - Physiology, pharmacology, and therapeutic relevance of urocortins in mammals: ancient CRF paralogs. AB - Urocortins, three paralogs of the stress-related peptide corticotropin-releasing factor (CRF) found in bony fish, amphibians, birds, and mammals, have unique phylogenies, pharmacologies, and tissue distributions. As a result and despite a structural family resemblance, the natural functions of urocortins and CRF in mammalian homeostatic responses differ substantially. Endogenous urocortins are neither simply counterpoints nor mimics of endogenous CRF action. In their own right, urocortins may be clinically relevant molecules in the pathogenesis or management of many conditions, including congestive heart failure, hypertension, gastrointestinal and inflammatory disorders (irritable bowel syndrome, active gastritis, gastroparesis, and rheumatoid arthritis), atopic/allergic disorders (dermatitis, urticaria, and asthma), pregnancy and parturition (preeclampsia, spontaneous abortion, onset, and maintenance of effective labor), major depression and obesity. Safety trials for intravenous urocortin treatment have already begun for the treatment of congestive heart failure. Further understanding the unique functions of urocortin 1, urocortin 2, and urocortin 3 action may uncover other therapeutic opportunities. PMID- 17083979 TI - Comment on "Immune reconstitution syndrome associated with parvovirus B19-induced pure red cell aplasia during highly active antiretroviral therapy" by Intalapaporn et al. PMID- 17083978 TI - Three new asymmetric trans-amine(azole)dichloridoplatinum complexes that overcome cisplatin resistance and their reactions with 5'-GMP. AB - Three new asymmetric platinum(II) complexes comprising an isopropylamine ligand trans to an azole ligand were synthesized and fully characterized by (1)H NMR, (195)Pt NMR, IR and elemental analysis. In addition the X-ray crystal structure of all three complexes was determined. The reaction kinetics of the complexes with DNA model base guanosine-5'-monophosphate (GMP) was studied, revealing reaction kinetics comparable to cisplatin. To gain insight in the complexes as potential antitumor agents, cytotoxicity assays were performed on a variety of human tumor cell lines. These assays showed the complexes all to possess cytotoxicity profiles comparable to cisplatin. Furthermore, the complexes largely retain their activity in a human ovarian carcinoma cell line resistant to cisplatin, A2780R, compared to the cisplatin sensitive parent cell line A2780. These results are of fundamental importance, illustrating how platinum complexes of trans geometry can show improved activity compared to cisplatin in both cisplatin sensitive and cisplatin resistant cell lines. PMID- 17083980 TI - A high throughput fluorescent assay for measuring the activity of fatty acid amide hydrolase. AB - Fatty acid amide hydrolase (FAAH) is the enzyme responsible for the rapid degradation of fatty acid amides such as the endocannabinoid anandamide. Inhibition of FAAH activity has been suggested as a therapeutic approach for the treatment of chronic pain, depression and anxiety, through local activation of the cannabinoid receptor CB1. We have developed a high throughput screening assay for identification of FAAH inhibitors using a novel substrate, decanoyl 7-amino-4 methyl coumarin (D-AMC) that is cleaved by FAAH to release decanoic acid and the highly fluorescent molecule 7-amino-4-methyl coumarin (AMC). This assay gives an excellent signal window for measuring FAAH activity and, as a continuous assay, inherently offers improved sensitivity and accuracy over previously reported endpoint assays. The assay was validated using a panel of known FAAH inhibitors and purified recombinant human FAAH, then converted to a 384 well format and used to screen a large library of compounds (>600,000 compounds) to identify FAAH inhibitors. This screen identified numerous novel FAAH inhibitors of diverse chemotypes. These hits confirmed using a native FAAH substrate, anandamide, and had very similar rank order potency to that obtained using the D-AMC substrate. Collectively these data demonstrate that D-AMC can be successfully used to rapidly and effectively identify novel FAAH inhibitors for potential therapeutic use. PMID- 17083981 TI - A novel method for monitoring the cell surface expression of heteromeric protein complexes in dispersed neurons and acute hippocampal slices. AB - The subunit composition of multimeric protein complexes is critical in determining their trafficking and functional properties. Despite there being multiple techniques to investigate the trafficking events of individual subunits there are currently limited means to monitor the trafficking properties of heteromeric protein complexes. Here, we combine surface biotinylation with co immunoprecipitation to monitor the cell surface expression of native, heteromeric AMPA receptor complexes. Using this method, we demonstrate that the surface levels of GluR1/2 and GluR2/3 complexes are reduced following NMDA-evoked long term depression (NMDA-LTD) in acute hippocampal slices. Finally, we discuss how this method can be adapted to monitor the cell surface expression of other heteromeric protein complexes. PMID- 17083982 TI - Spike detection in human muscle sympathetic nerve activity using the kurtosis of stationary wavelet transform coefficients. AB - The accurate assessment of autonomic sympathetic function is important in the diagnosis and study of various autonomic and cardiovascular disorders. Sympathetic function in humans can be assessed by recording the muscle sympathetic nerve activity, which is characterized by synchronous neuronal discharges separated by periods of neural silence dominated by colored Gaussian noise. The raw nerve activity is generally rectified, integrated, and quantified using the integrated burst rate or area. We propose an alternative quantification involving spike detection using a two-stage stationary wavelet transform (SWT) de noising method. The SWT coefficients are first separated into noise-related and burst-related coefficients on the basis of their local kurtosis. The noise related coefficients are then used to establish a threshold to identify spikes within the bursts. This method demonstrated better detection performance than an unsupervised amplitude discriminator and similar wavelet-based methods when confronted with simulated data of varying burst rate and signal to noise ratio. Additional validation on data acquired during a graded head-up tilt protocol revealed a strong correlation between the mean spike rate and the mean integrate burst rate (r=0.85) and burst area rate (r=0.91). In conclusion, the kurtosis based wavelet de-noising technique is a potentially useful method of studying sympathetic nerve activity in humans. PMID- 17083983 TI - Low-speed subcellular fractionation method for determining noxious stimulus evoked spinal neurokinin-1 receptor internalization. AB - Substance P release from nociceptive primary afferents activates post-synaptic neurokinin-1 (NK-1) receptors causing subsequent NK-1 receptor internalization. Fluorescent immunohistochemistry is typically used to quantify NK-1 receptor internalization, an indirect measure of substance P (SP) release. However, this technique entails several limitations that restrict its application. Using simple subcellular fractionation and immunoblotting methods, we demonstrate that intrathecal SP invokes a rapid and dose-dependent increase in dorsal horn cytoplasmic NK-1 receptors. We also show that hind paw compression and noxious thermal stimulation increase cytoplasmic NK-1 receptor, when compared to sham stimulations. Fluorescent immunohistochemistry confirmed that increases in cytoplasmic NK-1 corresponded with increased NK-1 receptor internalization. Herein, we report that low-speed centrifugation and Western immunoblotting provide NK-1 internalization results consistent with those obtained by more traditional methods. These data support previous findings demonstrating a role for spinal NK-1 receptors in nociceptive processing. PMID- 17083984 TI - A rapid method to measure beta-amyloid induced neurotoxicity in vitro. AB - Beta-amyloid (Abeta) is the primary protein component of senile plaques in Alzheimer's disease (AD) and is believed to be associated with neurotoxicity in the disease. Abeta-induced neurotoxicity is strongly dependent on its structure. The aggregation of the peptide from monomeric form to fibrils is a function of many variables including time. It is because of this dynamic nature of Abeta structure that there is a necessity for an in vitro toxicity assay that is rapid enough to eliminate or at least lower the possibility of Abeta structural changes during the time required for the assay. Here we describe a fast and sensitive method with which to assess Abeta-induced neurotoxicity in SH-SY5Y neuroblastoma cells. The method employs two-color flow cytometry using annexin-phycoerythrin, a marker for cell surface phosphatidylserine that typically indicates early stages of apoptosis, and 7-amino-actinomycin D, a membrane impermeant nucleic acid dye. We compare results using the two-color assay to those obtained using the propidium iodide toxicity assay and demonstrate comparability of results, but in 2h or less with the two-color assay as opposed to 24-48 h with the propidium iodide assay. The assay described could be a useful tool in evaluating the role of Abeta structure in biological activity of the peptide. PMID- 17083985 TI - Psychopathology in the young offspring of parents with bipolar disorder: a controlled pilot study. AB - Studies have suggested that the offspring of parents with bipolar disorder are at risk for a spectrum of psychopathology, but few have focused on children in the youngest age ranges or examined the impact of comorbid parental disorders. We utilized a pre-existing sample of young (mean age: 6.8 years) offspring of parents with bipolar disorder (n=34), of parents with panic or major depression (n=179), and of parents with neither mood or anxiety disorder (n=95). Children were assessed blindly to parental diagnoses using the Schedule for Affective Disorders and Schizophrenia-Epidemiologic version (K-SADS-E). Offspring of bipolar parents had significantly higher rates of disruptive behavior and anxiety disorders than offspring from both of the comparison groups, accounted for by elevated rates of ADHD and overanxious disorder. These comparisons were significant even when lifetime histories of the corresponding categories of comorbid disorders in the parents (disruptive behavior disorders and anxiety disorders) were covaried. In addition, offspring of bipolar parents had increased rates of bipolar I disorder, compared with psychiatric controls. Results support the hypotheses of elevated behavior, anxiety, and mood disorders among offspring at risk for bipolar disorder, and suggest that this psychopathology is already evident in early childhood. PMID- 17083986 TI - Maternal body burdens of methylmercury impair survival skills of offspring in Atlantic croaker (Micropogonias undulatus). AB - Methylmercury (MeHg), the organic form of mercury, bioaccumulates easily through the food chain. Fish in high trophic levels can accumulate substantial levels of MeHg and transfer it to their developing eggs. Here, the effects of maternally derived MeHg on the planktonic larval stage of Atlantic croaker were investigated. Adult Atlantic croaker were fed MeHg-contaminated food at three levels: 0, 0.05, and 0.1 mg kg(-1) day(-1) for 1 month. Fish were then induced to spawn and MeHg levels in the eggs were measured (0.04-4.6 ng g(-1)). Behavioral performance of exposed and control larvae was measured at four developmental stages: end of yolk absorption (yolk), end of oil absorption (oil), and 4 and 11 days after oil absorption (oil+4 and oil+11). Behaviors analyzed included survival skills related to foraging and predator evasion: routine behavior (rate of travel, active swimming speed, net-to-gross displacement ratio, and activity) and startle response to a visual and a vibratory stimulus (responsiveness, reactive distance, response distance, response duration, average response speed, and maximum response speed). Maternally transferred MeHg induced concentration dependent effects on survival skills. Statistical and simulation models applied to predict the ecological consequences of the behavioral effects suggested that maternal transfer of MeHg may substantially lower survival of planktonic stage larvae compared to unexposed larvae. These results also imply that larvae of top predatory fish species, such as blue marlin, may suffer mortality through maternal transfer of MeHg. PMID- 17083987 TI - Using an integrated approach to link biomarker responses and physiological stress to growth impairment of cadmium-exposed larval topsmelt. AB - In this study, we used an integrated approach to determine whether key biochemical, cellular, and physiological responses were related to growth impairment of cadmium (Cd)-exposed larval topsmelt (Atherinops affinis). Food intake (Artemia franciscana nauplii), oxygen consumption rates, apoptotic DNA fragmentation (TUNEL assay), and metallothionein (MT)-like protein levels, were separately measured in relation to growth of larval topsmelt aqueously exposed to sublethal doses of Cd for 14 days. Cadmium accumulation and concentrations of abundant metals were also evaluated in a subset of fish. Fish in the highest Cd treatments (50 and 100 ppb Cd) were smaller in final mean weight and length, and consumed fewer A. franciscana nauplii than control fish. Food intake was positively correlated with final weight of larval topsmelt in Cd and control treatments; food intake increased as final weight of the fish increased. Oxygen consumption rates were positively correlated with Cd concentration and mean oxygen consumption rates were inversely correlated with final mean weight of topsmelt; the smallest fish were found in the highest Cd treatment and were respiring at higher rates than control fish. Apoptotic DNA fragmentation was concentration-dependent and was associated with diminished growth. Apoptotic DNA fragmentation was elevated in the gill of fish exposed to 50 ppb Cd, and in the gut, gill, and liver of fish exposed to 100 ppb Cd. Metallothionein (MT)-like protein levels in fish from 100 ppb Cd treatments were significantly higher than those in other treatments. Oxygen consumption rates may have increased as a compensatory response to Cd exposure. However, it is likely that the energy produced was allocated to an increased metabolic demand due to apoptosis, MT synthesis, and changes in ion regulation. This diversion of energy expenditures could contribute to growth impairment of Cd-exposed fish. PMID- 17083988 TI - An unusual cause of shock in a patient with liver abscess. PMID- 17083989 TI - Synaptic plasticity modulates the spontaneous recovery of locomotion after spinal cord hemisection. AB - Several evidences have demonstrated that adult mammals could achieve a wide range of spontaneous sensory-motor recovery after spinal cord injury by means of various forms of neuroplasticity. In this study we evaluated the possibility that after low-thoracic spinal cord hemisection in the adult rat, significant hindlimb locomotor recovery could occur, and that this recovery may be driven, at least in part, by mechanisms of synaptic plasticity. In order to address these issues, we measured the expression levels of synapsin-I and brain-derived neurotrophic factor by Western blotting, at various time points after hemisection and correlated them with the motor performance on a grid walk test. Regression analysis showed that the expression of synapsin-I was strongly correlated with the spontaneous recovery of hindlimb locomotion (R=0.78). Conversely, neither the expression levels of synapsin-I nor the locomotor recovery were associated with the expression of brain-derived neurotrophic factor. Overall results indicate that after spinal cord hemisection, substantial recovery of hindlimb locomotion could occur spontaneously, and that synaptic plasticity within spinal circuitries below the level of the lesion, could be an important mechanism involved in these processes. PMID- 17083990 TI - High level expression of proteins using sequences from the ferritin heavy chain gene locus. AB - An expression vector has been generated using a gene highly expressed under conditions found in a typical fed-batch bioreactor process. The ferritin heavy chain (HC) gene exhibits higher levels of expression in the late stages of a fed batch bioreactor than in the early stages. This property was considered advantageous for an expression vector, since the maximal cell density would coincide with maximal expression. The rat ferritin HC genomic region was isolated and converted into an expression vector where large segments of 5' and 3' flanking regions were included in an attempt to recreate the same high level of expression in stably transfected cells. Expression from the resulting ferritin HC vector was compared to vectors containing the commonly used strong promoters, CMV IE, and SV40 early promoter/enhancer, in the generation of stable transfectants. The ferritin HC vector was able to generate cell lines with significantly higher expression levels than those under the control of the viral promoters. PMID- 17083991 TI - A bidirectional Tet-dependent promotor construct regulating the expression of E1A for tight control of oncolytic adenovirus replication. AB - Tight regulation of oncolytic adenoviruses (oAdV) represents an important requirement for their safe application. Here we describe a new doxycycline (Dox) dependent oAdV with a bidirectional expression cassette, which drives the expression of the reverse tetracycline-controlled transactivator (rtTA(s)-M2) from a lung tumor-specific promoter and, in the opposite direction, the expression of the adenoviral E1A gene from a second generation TetO(7) sequence linked to an isolated TATA box. In H441 lung cancer cells, this oAdV showed a strictly Dox-dependent E1A expression, adenoviral replication, cell killing activity and a 450-fold induction of progeny virus production. The virus could be shut off again by withdrawal of Dox and, in contrast to a control oAdV expressing E1A directly from the SP-B promoter, did not replicate in non-target cells. However, the absolute values of virus production and the cell killing activity in the presence of the inducer were still reduced as compared to the control oAdV. The results demonstrate, for the first time, Dox-dependent oAdV replication from a single adenoviral vector genome. Future improvement of the Dox-dependent E1A regulation cassette should lead to the generation of an oAdV well suited to meet the demands for a highly regulated and efficient oncolytic virus for in vivo applications. PMID- 17083992 TI - Enhancement of gene transfer activity mediated by mannosylated dendrimer/alpha cyclodextrin conjugate (generation 3, G3). AB - To enhance gene transfer activity of dendrimers, we prepared its conjugate (generation 3, G3) with alpha-cyclodextrin bearing mannose (Man-alpha-CDE conjugates) with various degrees of substitution of the mannose moiety (DSM5, 10, 13, 20) and compared their cytotoxicity and gene transfer activity, and elucidated the enhancing mechanism for the activity. Of the various carriers used here, Man-alpha-CDE conjugate (G3, DSM10) provided the highest gene transfer activity in NR8383, A549, NIH3T3 and HepG2 cells, being independent of the expression of mannose receptors. Gene transfer activity of Man-alpha-CDE conjugate (G3, DSM10) was not decreased by the addition of 10% serum in A549 cells. Cytotoxicity of the polyplex with Man-alpha-CDE conjugates (G3, DSM10) was not observed in A549 and NIH3T3 cells up to the charge ratio of 200/1 (carrier/pDNA). The gel mobility and particle size of polyplex with Man-alpha-CDE conjugate (G3, DSM10) were relevant to those with alpha-CDE conjugate (G3), but zeta-potential, DNase I stability, pDNA condensation of the former polyplex were somewhat different from those of the latter one. Cellular association of polyplex with Man-alpha-CDE conjugate (G3, DSM10) was almost comparable to that with dendrimer (G3) complex and alpha-CDE conjugate (G3). The addition of mannan and mannose attenuated gene transfer activity of Man-alpha-CDE conjugate (G3, DSM10) in A549 cells. Alexa-pDNA complex with TRITC-Man-alpha-CDE conjugate (G3, DSM10), but not the complex with TRITC-alpha-CDE conjugate (G3), was found to translocate to nucleus at 24 h after incubation in A549 cells. HVJ-E vector including mannan, but neither the vector alone nor the vector including dextran, suppressed the nuclear localization of TRITC-Man-alpha-CDE conjugate (G3, DSM10) to a striking degree after 24 h incubation in A549 cells. These results suggest that Man-alpha CDE conjugate (G3, DSM10) has less cytotoxicity and prominent gene transfer activity through not only its serum resistant and endosome-escaping abilities but also nuclear localization ability. PMID- 17083993 TI - Do the American College of Surgeons' "major resuscitation" trauma triage criteria predict emergency operative management? AB - STUDY OBJECTIVE: We wish to assess whether individual or collective American College of Surgeons' "major resuscitation" criteria accurately identify injured patients who receive emergency operative treatment. METHODS: In this observational secondary registry analysis of 8,289 consecutive trauma team activations during a 7.5-year period, we evaluated the test performance of 5 American College of Surgeons' major criteria in predicting emergency (within 1 hour) operative management by general (for adults) or pediatric (for children) surgeons. RESULTS: In adults, the individual major resuscitation criteria each predicted emergency operative management as follows (sorted from highest to lowest test performance): gunshot wounds to the neck or torso (likelihood ratio positive [LR+] 7.5; 95% confidence interval [CI] 6.2 to 9.1); confirmed hypotension (LR+ 5.3; 95% CI 4.0 to 7.1); interhospital transfers requiring blood transfusions (LR+ 4.6; 95% CI 2.6 to 8.2); respiratory compromise (LR+ 2.9; 95% CI 2.2 to 3.7), and Glasgow Coma Scale score less than 8 (LR+ 2.1; 95% CI 1.6 to 2.7). The collective strategy of using any of these 5 criteria exhibited a LR+ of 3.5 (95% CI 3.2 to 3.8), sensitivity 82% (95% CI 75% to 87%), and specificity 76% (95% CI 75% to 77%). Our findings in children were similar, but their precision was limited by the low baseline prevalence of emergency operative intervention. CONCLUSION: These 5 American College of Surgeons-mandated major resuscitation criteria vary several-fold in their individual ability to predict emergency operative management and collectively exhibit modest test characteristics for this purpose. Selective use of these criteria or revisions thereof could result in more efficient secondary trauma triage. Our results do not support the existing obligatory use of these criteria to maintain American College of Surgeons trauma center certification. PMID- 17083994 TI - Replacing hindsight with insight: toward better understanding of diagnostic failures. PMID- 17083995 TI - Fasting and emergency department procedural sedation and analgesia: a consensus based clinical practice advisory. AB - Emergency physicians frequently administer procedural sedation and analgesia to nonfasted patients; however, they currently have no specific guidelines to aid them in preprocedural risk stratification. We assembled a committee of leading emergency physician sedation researchers to develop a consensus-based clinical practice advisory for this purpose. Our goal was to create a tool to permit emergency physicians to identify prudent limits of sedation depth and timing in light of fasting status and individual patient risk factors. The advisory is not intended to assert a legal standard of practice or absolute requirement. It is expected that emergency physicians will at times appropriately deviate from it according to individualized judgment and unique clinical circumstances. PMID- 17083996 TI - Large acute cerebral hemorrhage presenting with pure alexia. AB - Alexia (the acquired inability to read) is an uncommon presenting complaint in the emergency department (ED). It is usually associated with a lesion in the brain located within the dominant hemisphere near the parieto-occipital junction, with some involvement of the splenium of the corpus callosum. Our review of the literature revealed multiple distinct causes for the finding of alexia, and a majority of the cases uncovered also involved agraphia (the acquired inability to write) and frequently visual field defects. We present the case of an otherwise healthy 35-year-old white man who arrived at our ED with the chief complaint of having difficulty reading. He was, however, able to write, and he exhibited no defects in his visual fields on gross testing in our ED. The patient was found to have a large, acute, intraparenchymal hemorrhage in the right posterior/inferior parietal cortex, very near the occipital lobe. We present this case, followed by a brief discussion, to heighten awareness of the complaint of alexia with or without agraphia as a possible presenting symptom of intracranial hemorrhage, or ischemic cerebrovascular accident. PMID- 17083997 TI - Research advances in procedural sedation and analgesia. PMID- 17083998 TI - Is informed consent required for the administration of intravenous contrast and similar clinical procedures? PMID- 17083999 TI - A novel range based QSAR study of human neuropeptide Y (NPY) Y5 receptor inhibitors. AB - A conventional QSAR study has been carried out using thermodynamic and other descriptors, on a set of arylsulfonamidomethylcyclohexyl derivatives as antagonists of potential obesity drug target human neuropeptide Y Y5 receptor. In addition, a novel range based method was applied to obtain a QSAR model so that the information contained in the compounds for which an approximate value instead of exact value of inhibitory activity was available could be included in the model. Analysis of models suggests that range based model is better in screening biologically active compounds from chemical library. The conventional model is able to predict activity accurately only for active compounds whereas the range based method is better in discriminating active and inactive compounds. PMID- 17084000 TI - Synthesis and biological evaluation of dialkylsubstituted maleic anhydrides as novel inhibitors of Cdc25 dual specificity phosphatases. AB - An efficient synthesis of dialkylsubstituted maleic anhydrides 1a-j is described. The inhibitory potential of these original anhydride derivatives was tested toward the three human isoforms A, B and C of dual specific phosphatases Cdc25. A micromolar range inhibition of Cdc25s was observed with the maleic anhydrides bearing simple alkyl side chains longer than C(9), to reach the optimal activity with a C(17) chain length. PMID- 17084001 TI - Immunomodulation produced by a green propolis extract on humoral and cellular responses of mice immunized with SuHV-1. AB - Despite recent technological advances in vaccine production, most the vaccines depend on the association with adjuvant substances. This work evaluated the adjuvant capacity of an ethanol extract of green propolis associated to inactivated Suid herpesvirus type 1 (SuHV-1) vaccine preparations. Mice inoculated with SuHV-1 vaccine plus aluminum hydroxide and 5mg/dose of propolis extract presented higher levels of antibodies when compared to animals that received the same vaccine without propolis. The use of SuHV-1 vaccine with propolis extract alone did not induce significant levels of antibodies, however it was able to increase the cellular immune response, evidenced by the increase in the expression of mRNA to IFN-gamma. Besides, propolis increased the percentage of protected animals against challenge with a lethal dose of SuHV-1. The effect of green propolis extract on the humoral and cellular immune responses may be exploited for the development of effective vaccines. PMID- 17084002 TI - Ex vivo infiltration of fibroblasts into the tendon deteriorates the mechanical properties of tendon fascicles but not those of tendon bundles. AB - BACKGROUND: After ligament reconstruction, mechanical deterioration of the grafted tendon is observed with revascularization and cellular infiltration. However, the effect of cellular infiltration on the mechanical properties of the tendon matrix has not been fully understood. METHODS: Cultured fibroblasts derived from the rabbit patellar tendon were seeded around an acellular rabbit patellar tendon that had undergone freeze-thaw treatment. At time-0, 3, and 6 weeks after seeding the cells, we evaluated cellular distribution in the tendon using a confocal laser microscope and the mechanical evaluations of the tendon fascicles and the tendon bundles. FINDINGS: The confocal laser microscopic analysis showed fibroblast infiltration ex vivo into the acellular tendon matrix. We could not find significant effects of the cellular infiltration on the tangent modulus of the tendon bundle, although the ex vivo cellular infiltration significantly reduced the modulus of the tendon fascicle. In addition, the tangent modulus of the incubated tendon without fibroblasts significantly decreased with time, particularly in the tendon bundle levels. INTERPRETATION: The findings of this study suggested that the effects of ex vivo cellular infiltration on the mechanical properties of the tendon bundles are relatively small, compared with its striking effect on the tendon fascicles. PMID- 17084003 TI - Bioaccumulation of trace elements in pelagic fish from the Western Indian Ocean. AB - Trace elements were analyzed in fish of commercial interest to determine their importance in marine systems of the Western Indian Ocean and their bioaccumulation patterns. The results are equivalent or lower than levels reported in ichthyofauna worldwide. Certain values of muscular Cd, Hg, Pb and Zn were, however, above thresholds for human consumption. Levels varied among tissues, species and fish length, but were seldom influenced by the nutritional condition of the fish, its gender and its reproductive status. Correlations between hepatic Hg and Se levels in Swordfish (r2=0.747) and Yellowfin Tunas (r2=0.226), and among metallothionein linking metals imply the existence of detoxification processes in these species. Level differences between fish from the Mozambique Channel and Reunion Island reflect differences of diets rather than differences of elemental availability in both environments. PMID- 17084004 TI - Perspectives regarding 50 years of research on effects of tropospheric ozone air pollution on US forests. AB - Tropospheric ozone (O(3)) was first determined to be phytotoxic to grapes in southern California in the 1950s. Investigations followed that showed O(3) to be the cause of foliar symptoms on tobacco and eastern white pine. In the 1960s, "X" disease of ponderosa pines within the San Bernardino Mountains was likewise determined to be due to O(3). Nearly 50 years of research have followed. Foliar O(3) symptoms have been verified under controlled chamber conditions. Studies have demonstrated negative growth effects on forest tree seedlings due to season long O(3) exposures, but due to complex interactions within forest stands, evidence of similar losses within mature tree canopies remains elusive. Investigations on tree growth, O(3) flux, and stand productivity are being conducted along natural O(3) gradients and in open-air exposure systems to better understand O(3) effects on forest ecosystems. Given projected trends in demographics, economic output and climate, O(3) impacts on US forests will continue and are likely to increase. PMID- 17084005 TI - Impact of elevated CO(2) and nitrogen fertilization on foliar elemental composition in a short rotation poplar plantation. AB - The experiment was carried out on a short rotation coppice culture of poplars (POP-EUROFACE, Central Italy), growing in a free air carbon dioxide enriched atmosphere (FACE). The specific objective of this work was to study whether elevated CO(2) and fertilization (two CO(2) treatments, elevated CO(2) and control, two N fertilization treatments, fertilized and unfertilized), as well as the interaction between treatments caused an unbalanced nutritional status of leaves in three poplar species (P. x euramericana, P. nigra and P. alba). Finally, we discuss the ecological implications of a possible change in foliar nutrients concentration. CO(2) enrichment reduced foliar nitrogen and increased the concentration of magnesium; whereas nitrogen fertilization had opposite effects on leaf nitrogen and magnesium concentrations. Moreover, the interaction between elevated CO(2) and N fertilization amplified some element unbalances such as the K/N-ratio. PMID- 17084006 TI - Chromium and nickel mobilization from a contaminated soil using chelants. AB - The mobilization of chromium and nickel from an industrial soil was investigated using two biodegradable chelants (citric acid and histidine), compared with a persistent one (ethylenediaminetetraacetic acid). Successive metal mobilizations were carried out in batch experiments. The main reactions involved were estimated by modeling the system with MINEQL+. For a single mobilization, citric acid was the most effective for Cr mobilization and EDTA for Ni. Their effectiveness could be explained by their ability to solubilize the mineral matrix and by the competition for the surfaces sites to desorb Cr(VI). Before and after the mobilizations, the distribution of metals was determined by a sequential extraction procedure. Only slight modifications were observed due to the low percentage of solubilized metal. A concentration of 0.05 mol L(-1) (citric acid and EDTA) allows a good compromise between metal mobilization and preservation of the soil mineral integrity. PMID- 17084007 TI - Assessing airborne pollution effects on bryophytes: lessons learned through long term integrated monitoring in Austria. AB - The study uses measured and calculated data on airborne pollutants, particularly nitrogen (ranges between 28 to 43kgN*ha(-1)*yr(-1)) and sulphur (10 to 18kgSO(4) S*ha(-1)*yr(-1)), in order to assess their long-term (1992 to 2005) effects on bryophytes at the UN-ECE Integrated Monitoring site 'Zobelboden' in Austria. Bryophytes were used as reaction indicators on 20 epiphytic plots using the IM monitoring method and on 14 terrestrial plots using standardised photography. The plots were recorded in the years 1992, 1993, 1998, and 2004/2005. Most species remained stable in terms of their overall population size during the observed period, even though there were rapid turnover rates of a large percentage of species on all investigated plots. Only a few bryophytes (Hypnum cupressiforme, Leucodon sciuroides) responded unambiguously to N and S deposition. Nitrogen deposition had a weak but significant effect on the distribution of bryophyte communities. However, the time shifts in bryophyte communities did not depend on total deposition of N and S. PMID- 17084008 TI - Safety evaluation of pigment containing Aspergillus carbonarius biomass in albino rats. AB - Aspergillus carbonarius, an ascomycetes fungus, is known to produce pectinase in solid-state fermentation. A mutant strain of A. carbonarius UV-10046 selected for temperature tolerance over produced polygalacturonase and during growth accumulated an yellow pigment in its biomass. Since the colored fungus suggested its application for food use, the freeze-dried biomass was evaluated to assess its safety in experimental animals. Acute and sub-acute toxicity studies were conducted on both sexes of albino rats. Feeding acute doses of A. carbonarius freeze-dried biomass at 0.5-5.0g/kg body weight to adult rats did not show any symptoms of toxicity or mortality of the rats. Similarly, dietary feeding of A. carbonarius at 0.25-2.0% level (w/w) for 14 weeks did not produce any significant changes in food intake or gain in body weight of the experimental rats compared to control rats. There were no significant differences in the relative weight of vital organs, hematological parameters, macroscopic and microscopic changes in vital organs and serum enzyme levels between the experimental and control groups. The results clearly showed that acute and sub-acute oral feeding of freeze-dried whole cells of A. carbonarius mutant for 14 weeks did not produce any toxic effects in male and female rats. PMID- 17084009 TI - A temporal study on the histopathological, biochemical and molecular responses of CCl(4)-induced hepatotoxicity in Cyp2e1-null mice. AB - Previous study using Cyp2e1-null mice showed that Cyp2e1 is required in CCl(4) induced liver injury at 24h, what remains unclear are the temporal changes in liver damage and the spectrum of genes involved in this process. We investigated the time-dependent liver changes that occurred at morphological, histopathological, biochemical and molecular levels in both Cyp2e1(+/+) and Cyp2e1(-/-) mice after treating with either corn oil or CCl(4) (1 ml/kg) for 2, 6, 12, 24 and 48 h. A pale orange colored liver, indicative of fatty infiltration, was observed in Cyp2e1(+/+) mice treated with CCl(4) for 24 and 48 h, while the Cyp2e1(+/+) mice treated with corn oil and Cyp2e1(-/-) mice treated with either corn oil or CCl(4) showed normal reddish brown colored liver. Ballooned hepatocytes with multiple vacuoles in their cytoplasm were observed in the livers of Cyp2e1(+/+) mice 24 and 48 h after treating with CCl(4). The levels of serum alanine aminotransferase and aspartate aminotransferase, markers for liver injury, were significantly higher at 12h, peaked at 24h and gradually decreased at 48 h after CCl(4) intoxication. In contrast, this kind of damage was not apparent in the Cyp2e1(-/-) mice treated with CCl(4). Altered expressions of genes related to liver cirrhosis, apoptosis, oxidative stress, xenobiotic detoxification, lipid metabolism, chemsensory signaling or tumorigenesis, structural organization, regeneration and inflammatory response were identified, and the time-dependent changes in expression of these genes were varied. Overall, the present study provides insights into the mechanism of CCl(4)-induced hepatotoxicity in animal models. PMID- 17084010 TI - Preventive effect of naringin on lipid peroxides and antioxidants in isoproterenol-induced cardiotoxicity in Wistar rats: biochemical and histopathological evidences. AB - This study was designed to evaluate the cardioprotective potential of naringin on lipid peroxides, enzymatic and nonenzymatic antioxidants and histopathological findings in isoproterenol (ISO)-induced myocardial infarction (MI) in rats. Subcutaneous injection of ISO (85 mg/kg) to male Wistar rats showed a significant increase in the levels of thiobarbituric acid reactive substances and lipid hydroperoxides in plasma and the heart and a significant decrease in the activities of superoxide dismutase, catalase, glutathione peroxidase and glutathione-S-transferase in the heart and the levels of reduced glutathione, vitamin C and vitamin E in plasma and heart and ceruloplasmin in plasma. Oral administration of naringin (10, 20 and 40 mg/kg, respectively) to ISO-induced rats daily for a period of 56 days showed a significant decrease in the levels of lipid peroxidative products and improved the antioxidant status by increasing the activities of antioxidant enzymes and nonenzymatic antioxidants. Histopathological findings of the myocardial tissue showed the protective role of naringin in ISO-induced rats. The effect at a dose of 40 mg/kg of naringin was more pronounced than that of the other two doses, 10 and 20mg/kg. The results of our study show that naringin possess anti-lipoperoxidative and antioxidant activity in experimentally induced cardiac toxicity. PMID- 17084011 TI - Melatonin has dose-dependent effects on folliculogenesis, oocyte maturation capacity and steroidogenesis. AB - Chemo and/or radiotherapy applied to young cancer patients most often have severe effects upon female fertility. Today, few options are available to protect ovarian function in females. However, these options are either ineffective, belong to the field of experimental research or/and are not applicable to all patients. Drugs that could protect the oocyte and its surrounding feeder cells from damage can be of great importance. Melatonin, being an important indirect antioxidant and a powerful direct free radical scavenger could be such a reagent. This paper reports the direct effects of different melatonin concentrations (range: 1 nM to 2 mM) on folliculogenesis and oogenesis of in vitro cultured mouse ovarian follicles. Early secondary mouse follicles were cultured in vitro for 12 days under different melatonin regimes. Every fourth day, survival rates were scored, follicles were morphologically evaluated and medium was collected for steroid analyses. On day 12, in vitro ovulation was induced by hCG/EGF. Eighteen hours later, oocytes were measured, oocyte maturation was evaluated and normality of spindle and chromosomes ascertained. Results obtained in this study indicated that 2mM melatonin is toxic. One mM negatively influenced oocyte maturation capacity. In the presence of 100 microM melatonin, androstenedione and progesterone were increased whereas estradiol was not influenced. Lower melatonin concentrations had no effect on the evaluated parameters. These data indicate an effect of melatonin on theca cell steroidogenesis. For prophylactic use, a dose of 10 microM could be suitable to reduce oxidative stress in cultured follicles. PMID- 17084012 TI - Successful resuscitation with thrombolysis of a patient suffering fulminant pulmonary embolism after recent intracerebral haemorrhage. AB - We report the life-saving administration of thrombolysis during cardiopulmonary resuscitation in a patient with recent intracerebral haemorrhage. A 53-year-old male with intracerebral haemorrhage was admitted to the intensive care unit. On the 24th day of treatment he suffered cardiac arrest with pulseless electrical activity. Transoesophageal echocardiography was performed during ongoing cardiopulmonary resuscitation. Thrombi in the right heart cavities with excessive right ventricular dysfunction confirmed the diagnosis of fulminant pulmonary embolism. Permanent restoration of a spontaneous rhythm was feasible only after administration of systemic thrombolysis with recombinant tissue plasminogen activator. Neurological examination and a computed tomogram of the brain did not show rebleeding. We conclude that under extreme circumstances absolute contraindications to thrombolysis should be weighed against the potential benefit. PMID- 17084013 TI - A fishy cause of sudden near fatal hypotension. AB - Seafood-borne illnesses are a common but under recognised source of morbidity. We report the case of an 80-year-old woman who presented to hospital after collapsing in a restaurant following lunch consisting of mackerel fish. A detailed food history and clinical exclusion helped diagnose the condition as scombroid poisoning. The patient made a complete recovery following antihistamine therapy. PMID- 17084014 TI - Nerve growth factor in treatment and pathogenesis of Alzheimer's disease. AB - The etiology of Alzheimer's disease (AD) is still unknown. In addition, this terrible neurodegenerative disease will increase exponentially over the next two decades due to longer lifespan and an aging "baby-boomer" generation. All treatments currently approved for AD have moderate efficacy in slowing the rate of cognitive decline in patients, and no efficacy in halting progression of the disease. Hence, there is an urgent need for new drug targets and delivery methods to slow or reverse the progression of AD. One molecule that has received much attention in its potential therapeutic role in AD is nerve growth factor (NGF). This review will demonstrate data from humans and animals which promote NGF as a potential therapeutic target by (1) outlining the hypothesis behind using NGF for the treatment of AD, (2) reviewing both the normal and AD altered signaling pathways and effects of NGF in the central nervous system (CNS), and (3) examining the results of NGF treatment obtained from animal models of AD and AD patients. PMID- 17084015 TI - A new continuous within-participants latent inhibition task: examining associations with schizotypy dimensions, smoking status and gender. AB - A within-participants latent inhibition task (LI: the retardation in learning that occurs if a stimulus has first been presented without consequence) was developed that produced robust LI, using both reaction time and number of correct responses as the dependent measures. Furthermore, associations were found between the preexposed stimulus and the schizotypy dimension of unusual experiences and smoking status. There were no significant relationships with the non-preexposed stimulus. This pattern of results was found using both reaction time and number of correct responses as the dependent measures which indicates that individuals who smoke and those high in unusual experiences show reduced LI. These findings indicate that the task may have sufficient sensitivity to be used in patients with schizophrenia. PMID- 17084016 TI - Dynamics of the nucleated polymerization model of prion replication. AB - The disease process for transmissible spongiform encephalopathies (TSEs), in one way or another, involves the conversion of a predominantly alpha-helical normal host-coded prion protein (PrP(C)) to an abnormally folded (predominantly beta sheet) protease resistant isoform (PrP(Sc)). Several alternative mechanisms have been proposed for this auto-catalytic process. Here the dynamical behavior of one of these models, the nucleated polymerization model, is studied by Monte Carlo discrete-event simulation of the explicit conversion reactions. These simulations demonstrate the characteristic dynamical behavior of this model for prion replication. Using estimates for the reaction rates and concentrations, time courses are estimated for concentration of PrP(Sc), PrP(Sc) aggregates, and PrP(C) as well as size distributions for the aggregates. The implications of these dynamics on protein misfolding cyclic amplification (PMCA) is discussed. PMID- 17084017 TI - A chemosensitivity test for superficial bladder cancer based on three-dimensional culture of tumour spheroids. AB - OBJECTIVES: Three-dimensional (3D) spheroids are a good model for studying in vitro chemosensitivity because they reproduce unicellular and multicellular mechanisms of drug resistance. We aimed to develop a chemosensitivity test for intravesical drugs and to also verify the effects of verapamil (VPM) and ciprofloxacin (CIPRO). METHODS: Cold cup biopsies from 40 superficial bladder tumours were taken, fragmented, and left in culture. 3D-spheroids were obtained and transferred into a 24 multiwell dish containing (1) wells 1-3: 1 mg/ml epirubicin (EPI); (2) 4-6: 1 mg/ml EPI+0.5 mg/ml VPM; (3) 7-9: 1 mg/ml adriamycin (ADR); (4) 10-12: 1 mg/ml thiotepa (THIO); (5) 13-15: 1 mg/ml mitomycin C (MMC); (6) 16-18: 1mg/ml EPI+0.2 mg/ml CIPRO; (7) 19-21: 0.2 mg/ml CIPRO; (8) 22-24: controls. Sensitivity was calculated by using the trypan blue assay. RESULTS: Evaluability of clinically relevant tests (G1-G2 lesions) was 84% (21 of 25 patients). MMC was the best agent (p<0.001) with mean sensitivity being 50%, followed by THIO (37%), EPI (7%), and ADR (3%). We found no significant difference (p=0.370) between CIPRO and the control, or between EPI+CIPRO and EPI alone (p=0.550). VPM markedly enhanced sensitivity to EPI compared with EPI alone (97% vs. 7%, p<0.001). CONCLUSIONS: Our assay allows determining sensitivity to several drugs in superficial bladder tumours. It might be used in clinical practice to select the best drug for each patient. It also has experimental utility in investigating the effect of new drugs or combinations. VPM reverted resistance to EPI. CIPRO showed no effect on bladder tumour spheroids. PMID- 17084018 TI - Overexpression of the mTOR alpha4 phosphoprotein activates protein phosphatase 2A and increases Stat1alpha binding to PIAS1. AB - Alpha4 phosphoprotein in the mTOR pathway is a prolactin (PRL)-downregulated gene product that interacts with the catalytic subunit of serine/threonine protein phosphatase 2A (PP2Ac) in rat Nb2 lymphoma cells. Transient overexpression of alpha4 in COS-1 cells inhibited PRL-inducible interferon-regulatory-1 (IRF-1) promoter activity, but the mechanism underlying this inhibition was not known. The present study showed a stable alpha4-PP2Ac complex that was not dissociated by rapamycin in COS-1 cells. Transient overexpression of alpha4 in COS-1 cells had no effect on endogenous PP2Ac protein levels but significantly increased PP2Ac carboxymethylation and PP2A activity as compared to controls. The increased PP2A activity was accompanied by decreased phosphorylation of eukaryotic initiation factor 4E-binding protein (4E-BP1) but had no effect on Stat phosphorylation. However, overexpressed alpha4 decreased arginine methylation of Stat1alpha and increased Stat1alpha binding to the Stat1alpha-specific inhibitor, PIAS1. In summary, ectopic alpha4 increased PP2A activity in COS-1 cells and this was accompanied by Stat1alpha hypomethylation and increased Stat1alpha-PIAS1 association. These events would inhibit Stat action and ultimately inhibit PRL inducible IRF-1 promoter activity. PMID- 17084019 TI - Transcriptional regulation of the FSH receptor: new perspectives. AB - The cell-surface receptor for the gonadotropin follicle-stimulating hormone (FSH) is expressed exclusively on Sertoli cells of the testis and granulosa cells of the ovary. FSH signal transduction through its receptor (Fshr) is critical for the timing and maintenance of normal gametogenesis in the mammalian gonad. In the 13 years since the gene encoding Fshr was first cloned, the mechanisms controlling its transcription have been extensively examined, but a clear understanding of what drives its unique cell-specificity remains elusive. Current knowledge of basal Fshr transcription highlights the role of an E-box in the proximal promoter which is bound by the basic helix-loop-helix transcription factors upstream stimulatory factor 1 (Usf1) and Usf2. Recent studies utilizing knockout mice and chromatin immunoprecipitation validated the importance of Usf to Fshr transcription and demonstrated a sexually dimorphic requirement for the Usf proteins to maintain normal Fshr expression. Studies have also shown that the promoter region itself is insufficient for appropriate Fshr expression in transgenic mice, indicating Fshr transcription depends on regulatory elements that lie outside of the promoter. Identification of such elements has been propelled by recent availability of genome sequence data, which facilitated studies using comparative genomics, DNase I hypersensitivity mapping, and transgenic analysis with large fragments of DNA. This review will focus on the current understanding of transcriptional regulatory processes that control expression of rat Fshr, including recent advances from our laboratory. PMID- 17084021 TI - Carbohydrate complexity and proportions of serum FSH isoforms in the male: lectin based studies. AB - FSH is synthesized and secreted in multiple molecular forms with different oligosaccharide structures which are needed for full expression of biological activity. GnRH and sex steroids modulate oligosaccharide structure and composition. In the present study we have assessed the carbohydrate complexity and proportion of circulating FSH isoforms during puberty, aging and after androgen administration to pubertal anorchid boys. Preparative isoelectrofocusing and lectin chromatography were used to isolate FSH isoforms on the basis of charge and internal carbohydrate complexity. Differences in sialic acid content and a progressive increase of isoforms bearing highly branched oligosaccharides were found during puberty. Less acidic, more bioactive FSH isoforms, secreted at mid-puberty may modulate important maturational events in the Sertoli cell population. Androgen administration to pubertal anorchid boys favoured the secretion of this type of isoforms. In adult men, the predominance of FSH isoforms bearing complex type oligosaccharides remained unchanged until very advanced age. These results show that the predominance of FSH isoforms bearing fully processed oligosaccharides in circulation may contribute to the development and maintenance of seminiferous epithelium function in men. PMID- 17084020 TI - Patterns of LHbetacf among women in health and disease. AB - Urine based gonadotropin assays provide a practical means of analyzing hormone secretion patterns. While research protocols have revealed pulsatile patterns of gonadotropins such as LH in the blood, these assays are of limited clinical use since daily venipuncture sampling is not feasible outside of a research environment. However, collection of several urine samples provides a method to achieve the same visualization of gonadotropin patterns in patients using a convenient and generally applicable technique based on analysis of the highly stable hLHbetacf for monitoring LH and hCGbetacf for monitoring pituitary hCG. We demonstrated that two different sampling techniques for analyzing these gonadotropin metabolites yielded the same information on their excretory patterns, either sampling of spot urines or collecting first morning void urines for several days. Next, we studied the core excretory patterns in several populations: menstruating and postmenopausal women from the general population, and two populations of women from a fertility center, one of which had polycystic ovaries (PCO). The PCO population was also subdivided into those with and without insulin resistance (IR). It was found that our hLHbetacf assay did not measure the form of the LH core (v-hLHbetacf) produced in subjects who were homozygous for a variant form of LH (v-LH). None of our patients tested were homozygous for the variant form of LH. It was also found that in most non-PCO (NPCO) patients, the hLHbetacf peak lasted for 7-9 days while among the PCO patients this peak frequently lasted for less than 7 days and an erratic pattern tended to appear. The overall differences in patterns between the PCO and NPCO patients were confirmed by spectral statistical methods. The prevalence of certain characteristic hLHbetacf patterns may be higher among women with PCO with a more severe clinical presentation. Use of urinary analysis of gonadotropin metabolites, especially hLHbetacf, may supplement subjective ultrasound studies with more sensitive biochemical measurements. PMID- 17084022 TI - SKY analysis of childhood neural tumors and cell lines demonstrates a susceptibility of aberrant chromosomes to further rearrangements. AB - Malignant solid tumors are commonly characterized by a large number of complex structural and numerical chromosomal alterations, which often reflect the level of genomic instability and can be associated with disease progression. The aim of this study was to evaluate whether chromosomes that harbor primary aberrations have a higher susceptibility to accumulate further alterations. We used spectral karyotyping (SKY), to compare the individual chromosomal instability of two chromosome types: chromosomes that have a primary aberration and chromosomes without an aberration, in 13 primary childhood neural tumors and seven cell lines. We found that chromosomes that contain a primary aberration are significantly (p-value<0.001) more likely to gain further structural rearrangements or to undergo numerical changes (22.6%, 36 of 159 chromosomes) than chromosomes with no initial aberration (4.9%, 54 of 1099 chromosomes). These results are highly suggestive that aberrant chromosomes in solid tumors have a higher susceptibility to accumulate further rearrangements than "normal" chromosomes. PMID- 17084023 TI - Preparation and photocatalytic properties of YVO4 nanopowders. AB - YVO(4) photocatalysts with different grain sizes were obtained by annealing the YVO(4) nanopowders synthesized via microwave irradiation. The products were characterized by X-ray diffractometer (XRD), scanning electron microscope (SEM), transmission electron microscope (TEM), surface area and porosity analyzer, and ultraviolet-visible (UV-vis) spectrophotometer. Photocatalytic measurements showed that the YVO(4) nanopowders with grain size of about 5 nm possess superior photocatalytic properties in decolorization of methyl orange solution. PMID- 17084024 TI - The use of sulphuric acid-carbonization products of sugar beet pulp in Cr(VI) removal. AB - A carbon rich adsorbent prepared from the reaction of sugar beet pulp with sulphuric acid and gas formed during carbonization process have been studied for Cr(VI) removal from aqueous solutions. The SO(2) rich gas was shown to be an excellent Cr(VI) reductant. The equilibrium and kinetic studies were conducted by using the carbonaceous adsorbent derived from sugar beet pulp. The lower pH favoured Cr(VI) adsorption but substantial Cr(VI) reduction was observed. The Langmuir and Freundlich isotherm models were applied and the Langmuir model best fit the equilibrium isotherm data. The maximum adsorption capacity of chromium calculated from Langmuir isotherm is about 24 mgg(-1) for 25 degrees C. The adsorption of Cr(VI) is an endothermic process and follows the pseudo-second order rate kinetics. The sulphuric acid-carbonization is an economical method for particularly chromium removal because the gas generated during carbonization exhibits good Cr(VI) reduction properties and carbonaceous material obtained is an efficient Cr(VI) adsorbent. PMID- 17084025 TI - Effect of metal doping into Ce0.5Zr0.5O2 on photocatalytic activity of TiO2/Ce0.45Zr0.45M0.1OX (M=Y, La, Mn). AB - The paper demonstrates that the photocatalytic activity of TiO2 towards the decomposition of gaseous benzene in a batch reactor can be greatly improved by loading TiO2 on the surface of CeO(2)-ZrO(2). The research investigates the effects of three metals doping into Ce(0.5)Zr(0.5)O(2) on photocatalytic activity of TiO2/Ce(0.45)Zr(0.45)M(0.1)O(X) (M=Y, La, Mn). The prepared photocatalysts were characterized by BET, XRD, UV-vis diffuse reflectance and XPS analyses. BET surface area of TiO2/Ce(0.45)Zr(0.45)M(0.1)O(X) (M=Y, La, Mn) is smaller than that of Ce(0.5)Zr(0.5)O(2). XRD results reveal that the deposited titania is highly dispersed as in the CeO(2)-ZrO(2) matrix, doping M in the CeO(2)-ZrO(2) lattice causes the changing of lattice space and the diffraction peaks shift to higher 2theta position. Among these four catalysts, the band gap value of TiO(2)/Ce(0.45)Zr(0.45)La(0.1)O(X) is the lowest. The binding energy value of Ti 2p(3/2) of four catalysts transfers to a lower value. The order of photocatalytic activity is TiO2/Ce(0.45)Zr(0.45)La(0.1)O(X)>TiO2/Ce(0.45)Zr(0.45)Y(0.1)O(X)>TiO2/Ce(0.45)Zr( .45)Mn(0.1)O(X)>TiO2/Ce(0.5)Zr(0.5)O(2)>TiO2. The proposed mechanism is of electron transfer and the stronger absorption in the region 210-400 nm. PMID- 17084026 TI - Electroacupuncture stimulates the expression of prolactin-releasing peptide (PrRP) in the medulla oblongata of ovariectomized rats. AB - Electroacupuncture (EA) in reproductive medicine has become established in Western medicine as a therapy over the last decade. EA performs a variety of neuromodulatory functions in the central nervous system (CNS). Prolactin releasing peptide (PrRP) is a neuropeptide identified as an endogenous ligand for the orphan G protein-coupled receptor hGR3. PrRP can affect the function of hypothalamus-pituitary-ovary axis (HPOA) and hypothalamus-pituitary-adrenal axis (HPAA). The present study was undertaken to characterize the effect of EA on the expression of PrRP in the medulla oblongata in ovariectomized (OVX) rats by immunohistochemistry and reverse transcription-polymerase chain reaction (RT PCR). In addition, estrogen (E2) levels were detected by radioimmunoassay (RIA). The results suggest that EA significantly increase the blood level of E2 and the expression of PrRP in the medulla oblongata of OVX rats. The number of PrRP immunoreactive (ir) neurons was higher in the group ovariectomized with EA than that in the OVX group. The numbers of PrRP-ir neurons in intact (INT) and intact with EA (INT+EA) were not significantly different between the two groups. The expression of PrRP mRNA was increased in the OVX+EA group than that in the OVX group. These results suggest that the mechanism that EA improved reproductive disorders induced by ovariectomy in rats is related to the modulation of the blood E2 level and the expression of PrRP in the medulla oblongata. PMID- 17084027 TI - The sound of stress: blunted cortisol reactivity to psychosocial stress in tinnitus sufferers. AB - Clinical observations suggest that tinnitus is modulated by stress. However, there is little empirical data to support the link between stress and tinnitus. In this study, we measured the stress hormone cortisol to examine the reactivity of the hypothalamic-pituitary-adrenal (HPA) axis in tinnitus participants as well as in healthy controls without tinnitus. Eighteen participants with tinnitus and 18 controls without tinnitus were exposed to the Trier Social Stress Task and cortisol sampling and subjective ratings were obtained at regular intervals. Tinnitus participants displayed a blunted cortisol response to psychosocial stress, in comparison with healthy controls who had a typical cortisol release about 30 min after the beginning of the experiment. The blunted cortisol response displayed by the tinnitus participants suggests that they have an anomaly along the HPA axis. Their cortisol response is similar to that found in other bodily stress-related diseases and thus suggests that tinnitus is related to stress. However, tinnitus intensity might not be modulated by stress in a concurrent manner. PMID- 17084028 TI - Efficacy of biofeedback for migraine: a meta-analysis. AB - In this article, we meta-analytically examined the efficacy of biofeedback (BFB) in treating migraine. A computerized literature search of the databases Medline, PsycInfo, Psyndex and the Cochrane library, enhanced by a hand search, identified 86 outcome studies. A total of 55 studies, including randomized controlled trials as well as pre-post trials, met our inclusion criteria and were integrated. A medium effect size (d =0.58, 95% CI=0.52, 0.64) resulted for all BFB interventions and proved stable over an average follow-up phase of 17 months. Also, BFB was more effective than control conditions. Frequency of migraine attacks and perceived self-efficacy demonstrated the strongest improvements. Blood-volume-pulse feedback yielded higher effect sizes than peripheral skin temperature feedback and electromyography feedback. Moderator analyses revealed BFB in combination with home training to be more effective than therapies without home training. The influence of the meta-analytical methods on the effect sizes was systematically explored and the results proved to be robust across different methods of effect size calculation. Furthermore, there was no substantial relation between the validity of the integrated studies and the direct treatment effects. Finally, an intention-to-treat analysis showed that the treatment effects remained stable, even when drop-outs were considered as nonresponders. PMID- 17084029 TI - Occurrence of Babesia felis and Babesia leo in various wild felid species and domestic cats in Southern Africa, based on reverse line blot analysis. AB - Reverse line blot (RLB) is a hybridization assay that can be used to detect various blood parasites and differentiate between them. Results, using the RLB, showed that Babesia felis and Babesia leo occurred as single or mixed infections in various felid species, but most frequently in domestic cats and lions, respectively. Prevalence of infection in free-ranging cheetahs in Namibia was low (7, 5%), whereas 50% of free-ranging lions in South Africa and Swaziland were infected. A large number (52, 9%) of samples tested positive only for Babesia, neither B. felis nor B. leo. This could be an indication of at least one further, as yet undescribed, Babesia species in felids. PMID- 17084030 TI - Suprathel-an innovative, resorbable skin substitute for the treatment of burn victims. AB - Autologous split skin grafts are the most reliable method for closing third degree burns. Under this scheme, donor sites as well as second degree burns under conservative treatment, however, would benefit from rapid wound closure. For this treatment, biological as well as synthetic materials are available. For the improvement of these materials, primary goals are pain reduction and easy handling in the absence of biological risk. From a synthetic copolymer mainly based on DL-lactic acid a new skin substitute was developed, marketed as Suprathel. Within the scope of a bicentric study Suprathel was compared versus paraffin gauze intraindividually applied on split skin donor sites. Wound pain was measured on the Visual Pain Analog Scale over a period of 10 days as the critical criterion. Accordingly Suprathel versus Omiderm were compared on second degree burns (degree 2a, partial thickness burns). In both study parts, Suprathel significantly reduced pain. Its easy handling was superior compared to other materials. The Suprathel membrane adhered rapidly to the wound thus protecting against infections and promoting wound healing. No allergic reactions were observed. The ability of the material to resorb ensured pain-free removal after complete healing of the wound. PMID- 17084031 TI - Burns in Turkish children and adolescents: nine years of experience. AB - AIM: The aim of this study was to describe information about burns that occur in children and adolescents in Turkey. PATIENTS AND METHODS: The subjects were 362 patients whom were younger than 18 years who were treated at 3 burn centers in 2 different regions of Turkey between 1997 and 2005. The data collected for each case were age, gender, place of residence, cause and extent of burn, body sites affected, environment in which the injury occurred, interval from injury to arrival at a burn center, hospitalization status (inpatient versus outpatient), surgical treatment, and mortality. RESULTS: The 362 patients comprised 35.5% of all 1021 burn victims admitted during the study period. There were 183 boys and 179 girls (ratio 1:0.98) and the mean total body surface area burned was 17.7+/ 16.5%. The highest proportion of patients were in the 1-6 years age group. Non bath (not immersed) hot water scalding (216 cases, 59.7%) was the leading burn cause. The most common environment in which burn injury occurred was the home. The trunk was the body site most frequently affected (62.7%). 241 (66.6%) subjects lived in urban environments and 121 (33.4%) lived in rural areas. 171 patients (47.2%) were taken directly to the burn units, whereas the others (52.8%) were referred from other medical centers. 124 (34.3%) subjects were treated as outpatients and 238 (65.7%) were hospitalized. The overall mortality rate was 8.6% (31 deaths). Of the 238 inpatients, 92 (38.7%) were treated with daily dressings only, 128 (53.8%) required debridement, and 75 (31.5%) needed both debridement and grafting. CONCLUSION: Every country needs a nationwide public education system that is aimed at preventing burns and ensuring that burn victims receive proper first aid and age-appropriate, specialized burn care. PMID- 17084032 TI - Death following severe burns and claims of medical negligence clarified. PMID- 17084033 TI - Bisphosphonate-related jaw necrosis--severe complication in maxillofacial surgery. AB - Bisphosphonates are used as potent inhibitors in metastatic bone lesions. They can reduce skeletal burden and prevent bony metastases. They are integral in the treatment of some tumours like breast cancer, prostate cancer and multiple myeloma. As a side effect, these drugs also may cause severe jaw necrosis. Twenty four patients with bisphosphonate-related jaw necrosis were analyzed in a clinical study. These necroses mostly appeared after administration of aminobisphosphonates. Recurrent avascular necroses were found after changing from Pramidronate to Zoledronate. All patients were treated by resection of necrotic bone. Repeated surgical interventions were required with about 25% of the patients. The management of patients with bisphosphonate-related jaw necrosis remains extremely difficult and includes surgical procedures as well as the eradicating of the necrotic bone including antibiotic therapy. The prevention of such complications consists in a minimization of dental surgical interventions and an avoidance of ulcers by dental prosthesis. PMID- 17084034 TI - Thrombin induces striatal neurotoxicity depending on mitogen-activated protein kinase pathways in vivo. AB - Intracerebral hemorrhage represents stroke characterized by formation and expansion of hematoma within brain parenchyma. Blood-derived factors released from hematoma are considered to be involved in poor prognosis of this disorder. We previously reported that thrombin, a blood-derived serine protease, induced cytotoxicity in the cerebral cortex and the striatum in organotypic slice cultures, which depended on mitogen-activated protein kinase (MAPK) pathways. Here we investigated the mechanisms of thrombin cytotoxicity in the striatum in vivo. Thrombin microinjected into the striatum of adult rats induced neuronal death and microglial activation around the injection site. Neuronal loss without any sign of nuclear fragmentation was observed as early as 4 h after thrombin injection, which was followed by gradual neuronal death exhibiting nuclear fragmentation. Thrombin-induced damage assessed at 72 h after injection was partially but significantly reduced by concomitant administration of inhibitors of MAPK pathways. Activation of extracellular signal-regulated kinase (ERK) and p38 MAPK in response to thrombin was verified by Western blot analysis. Moreover, phosphorylated ERK and p38 MAPK were localized prominently in reactive microglia, and inhibition of microglial activation by minocycline attenuated thrombin induced damage, suggesting that reactive microglia were responsible for thrombin induced neuronal death. Thus, MAPK pathways and microglial activation may serve as therapeutic targets of pathogenic conditions associated with hemorrhagic stroke. PMID- 17084035 TI - Anti-apoptotic effect of granulocyte-colony stimulating factor after focal cerebral ischemia in the rat. AB - We investigated the molecular mechanisms of the anti-apoptotic properties of granulocyte-colony stimulating factor (G-CSF) on neurons and whether G-CSF affects glial cell survival following focal cerebral ischemia in rats. Sprague Dawley rats were subjected to a transient 90 min middle cerebral artery occlusion (MCAO) by the intraluminal occlusion technique. Rats were treated with either a single dose of G-CSF (50 microg/kg, s.c.) at the onset of reperfusion or G-CSF (50 microg/kg body weight, s.c.) was administered starting at the onset of reperfusion and followed by the administration of the same dose per day for an additional 2 days. Brains were harvested either 24 h, 72 h or 2 weeks after reperfusion for assays of infarct volume, immunohistological studies and Western blot analysis for phosphorylated signal transducer and activator of transcription 3 (pSTAT3), Pim-1, bcl-2, Bax, cytochrome c, cellular inhibitor of apoptosis protein 2 (cIAP2), and cleaved caspase-3 levels. G-CSF significantly reduced infarct volume and ameliorated the early neurological outcome. G-CSF treatment significantly up-regulated pSTAT3, Pim-1, bcl-2 expression, and down-regulated cytochrome c release to the cytosol, Bax translocation to the mitochondria, and cleaved caspase-3 levels in neurons. The activation of the STAT3 pathway was accompanied by increased cIAP2 expression in glial cells. After MCAO, G-CSF treatment increased both neuronal and glial survival by effecting different anti apoptotic pathways which reflects the multifactorial actions of this drug. These changes were associated with remarkable improvement in tissue preservation and behavioral outcome. PMID- 17084036 TI - The effect of amphetamine analogs on cleaved microtubule-associated protein-tau formation in the rat brain. AB - The present study quantified the cleaved form of the microtubule-associated protein tau (cleaved MAP-tau, C-tau), a previously demonstrated marker of CNS toxicity, following the administration of monoamine-depleting regimens of the psychostimulant drugs amphetamine (AMPH), methamphetamine (METH), +/-3,4 methylenedioxymethamphetamine (MDMA), or para-methoxyamphetamine (PMA) in an attempt to further characterize psychostimulant-induced toxicity. A dopamine (DA) depleting regimen of AMPH produced an increase in C-tau immunoreactivity in the striatum, while a DA- and serotonin (5-HT)-depleting regimen of METH produced an increase in the number of C-tau immunoreactive cells in the striatum and CA2/CA3 and dentate gyrus regions of the hippocampus. MDMA and PMA, two psychostimulant drugs that produce selective 5-HT depletion in the striatum, had no effect on C tau immunoreactivity in the striatum or hippocampus. Furthermore, 5,7 dihydroxytryptamine (5,7-DHT), an established 5-HT selective neurotoxin, did not produce an increase in C-tau immunoreactivity. Dual fluorescent immunocytochemistry with antibodies to glial fibrillary acidic protein (GFAP) and C-tau indicated that C-tau immunoreactivity was present in astrocytes, not neurons, suggesting that increased C-tau may be an alternative indicator of reactive gliosis. The present results are consistent with previous findings that the DA-depleting psychostimulants AMPH and METH produce reactive gliosis whereas the 5-HT-depleting drugs MDMA and PMA, as well as the known 5-HT selective neurotoxin 5,7-DHT, do not produce an appreciable glial response. PMID- 17084037 TI - The role of poly(ADP-ribose) polymerase-1 in CNS disease. AB - Poly(ADP-ribose) polymerase-1 (PARP-1) is a nuclear enzyme that contributes to both neuronal death and survival under stress conditions. PARP-1 is the most abundant of several PARP family members, accounting for more than 85% of nuclear PARP activity, and is present in all nucleated cells of multicellular animals. When activated by DNA damage, PARP-1 consumes nicotinamide adenine dinucleotide (NAD+) to form branched polymers of ADP-ribose on target proteins. This process can have at least three important consequences in the CNS, depending on the cell type and the extent of DNA damage: 1) Poly(ADP-ribose) formation on histones and on enzymes involved in DNA repair can prevent sister chromatid exchange and facilitate base-excision repair; 2) poly(ADP-ribose) formation can influence the action of transcription factors, notably nuclear factor kappaB, and thereby promote inflammation; and 3) extensive PARP-1 activation can promote neuronal death through mechanisms involving NAD+ depletion and release of apoptosis inducing factor from the mitochondria. PARP-1 activation is thereby a key mediator of neuronal death during excitotoxicity, ischemia, and oxidative stress, and PARP-1 gene deletion or pharmacological inhibition can markedly improve neuronal survival in these settings. PARP-1 activation has also been identified in Alzheimer's disease and in experimental allergic encephalitis, but the role of PARP-1 in these disorders remains to be established. PMID- 17084038 TI - A novel splice site mutation in the Cockayne syndrome group A gene in two siblings with Cockayne syndrome. AB - Cockayne syndrome (CS) is mainly caused by mutations in the Cockayne syndrome group A or B (CSA or CSB) genes which are required for a sub-pathway of nucleotide excision repair entitled transcription coupled repair. Approximately 20% of the CS patients have mutations in CSA, which encodes a 44 kDa tryptophane (Trp, W) and aspartic acid (Asp, D) amino acids (WD) repeat protein. Up to now, nine different CSA mutations have been identified. We examined two Somali siblings 9 and 12 years old with clinical features typical of CS including skin photosensitivity, progressive ataxia, spasticity, hearing loss, central and peripheral demyelination and intracranial calcifications. Molecular analysis showed a novel splice acceptor site mutation, a G to A transition in the -1 position of intervening sequence 6 (g.IVS6-1G>A), in the CSA (excision repair cross-complementing 8 (ERCC8)) gene. IVS6-1G>A results in a new 28 amino acid C terminus and premature termination of the CSA protein (G184DFs28X). A review of the CSA protein and the 10 known CSA mutations is also presented. PMID- 17084039 TI - Activation of protein kinase B/Akt in the periphery contributes to pain behavior induced by capsaicin in rats. AB - Protein kinase B (PKB/Akt) is a member of the second-messenger regulated subfamily of protein kinases. It is implicated in signaling downstream of growth factors, insulin receptor tyrosine kinases and phosphoinositide 3-kinase (PI3K). Current studies indicate that nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and PI3K help mediate inflammatory hyperalgesia. However, little is known about the role of PKB/Akt in the nociceptive system. In this study, we investigated whether PKB/Akt in primary sensory neurons is activated after noxious stimulation and contributes to pain behavior induced in rats by capsaicin. We demonstrated that phospho-PKB/Akt (p-PKB/Akt) is increased in dorsal root ganglia (DRG) at 5 min after intradermal injection of capsaicin. p PKB/Akt is distributed predominantly in small- and medium-sized DRG cells. After capsaicin injection, p-PKB/Akt (473) is colocalized with isotectin-B4 (IB4), tyrosine kinase A (TrkA), and calcitonin gene-related peptide (CGRP). Furthermore, most transient receptor potential vanilloid type 1 (TRPV1) positive DRG neurons double label for p-PKB/Akt. Behavioral experiments show that intradermal injection of a PI3K (upstream of PKB/Akt) inhibitor, wortmannin, dose dependently inhibits the changes in exploratory behavior evoked by capsaicin injection. The PKB/Akt inhibitor, Akt inhibitor IV, has the same effect. The results suggest that the PKB/Akt signaling pathway in the periphery is activated by noxious stimulation and contributes to pain behavior. PMID- 17084040 TI - Psychoneuroendocrinology of anorexia nervosa. AB - It is suggested that the symptoms of anorexia nervosa are physiological responses to starvation. There is no evidence of a neural or non-neural dysfunction that predisposes women for anorexia nervosa and the endocrine and psychological consequences of starvation are reversed once patients have re-learnt how to eat and regained a normal body weight. Because variability in the supply of food may be a common evolutionary condition, it is more likely that body weight is variable than constant in normal circumstances. The role of the neuroendocrine system in times of feast and famine is to allow the individual to adopt behavioral strategies as needed rather than maintaining body weight homeostasis. Treatment of anorexic patients should aim at reducing their high level of physical activity in order to facilitate eating. PMID- 17084041 TI - Sensory modality and time perception in children and adults. AB - This experiment investigated the effect of signal modality on time perception in 5- and 8-year-old children as well as young adults using a duration bisection task in which auditory and visual signals were presented in the same test session and shared common anchor durations. Durations were judged shorter for visual than for auditory signals by all age groups. However, the magnitude of this modality difference was larger in the children than in the adults. Sensitivity to time was also observed to increase with age for both modalities. Taken together, these two observations suggest that the greater modality effect on duration judgments for the children, for whom attentional abilities are considered limited, is the result of visual signals requiring more attentional resources than are needed for the processing of auditory signals. Within the framework of the information processing model of Scalar Timing Theory, these effects are consistent with a developmental difference in the operation of the "attentional switch" used to transfer pulses from the pacemaker into the accumulator. Specifically, although timing is more automatic for auditory than visual signals in both children and young adults, children have greater difficulty in keeping the switch in the closed state during the timing of visual signals. PMID- 17084042 TI - Flounder and fugu have a single lefty gene that covers the functions of lefty1 and lefty2 of zebrafish during L-R patterning. AB - The lefty gene encodes a member of the TGF-beta superfamily that regulates L-R axis formation during embryogenesis via antagonistic activity against Nodal, another TGF-beta superfamily member. Both mouse and zebrafish have two lefty genes, lefty1 and lefty2. Interestingly, the expression domains of mouse and zebrafish lefty are different from one another. At present, the orthology and functional diversity of the mouse and zebrafish lefty genes are not clear. Here, we report that flounder and two fugu species, Takifugu and Tetraodon, have a single lefty gene in their genomes. In addition, we provide evidence that the mouse lefty genes were duplicated on a single chromosome but the zebrafish lefty genes arose from a whole-genome duplication that occurred early in the divergence of ray-finned fishes. These independent origins likely explain the difference in the expression domains of the mouse and zebrafish lefty gene pairs. Furthermore, we found that the duplication corresponding to the zebrafish lefty2 gene was lost from the fugu genome, suggesting that loss of lefty2 in the fugu/flounder lineage occurred after its divergence from the zebrafish lineage. During L-R patterning, the single lefty gene of flounder covers two expression domains, the left side of the dorsal diencephalon and the left LPM, which are regulated separately by lefty1 and lefty2 in zebrafish. We infer that the lefty genes of the ray-finned fishes and mammals underwent independent gene duplication events that resulted in independent regulation of lefty expression. PMID- 17084043 TI - Comparison of two selective media for the recovery, isolation, enumeration and differentiation of Rhodococcus equi. AB - The use of selective media to facilitate the isolation of Rhodococcus equi from environmental and clinical samples has aided studies of the ecology of R. equi and the epidemiology of disease caused by R. equi. Here, we compared the efficacy of two selective media (NANAT and modified CAZ-NB) for the recovery of six defined strains of R. equi and for the isolation and enumeration of both avirulent and virulent R. equi from 60 paired soil samples from horse farms using colony blotting and DNA hybridisation. No difference was found between the two media in the recoverability of defined strains of R. equi or the proportion of soil cultures positive for R. equi or virulent R. equi. NANAT medium was significantly less inhibitory of bacterial growth from soil culture compared to mCAZ-NB (P = 0.001), but there was no difference between the media in the number of R. equi colonies recovered. Soil cultured on mCAZ-NB medium yielded a significantly greater number of virulent R. equi colonies than NANAT (P = 0.03). The proportion of R. equi that were virulent in soil cultures on mCAZ-NB (32%) was more than three times that seen in cultures on NANAT (9%). Thus modified CAZ NB appeared to be a better selective media for studies where the optimal recovery of virulent R. equi is required, such as in studies of the gastrointestinal carriage of virulent R. equi and of subclinically infected foals. PMID- 17084044 TI - Analysis of world strains of Anaplasma marginale using major surface protein 1a repeat sequences. AB - Anaplasma marginale is a tick-borne pathogen of cattle that causes the disease bovine anaplasmosis worldwide. Major surface proteins (MSPs) are involved in host pathogen and tick-pathogen interactions and have been used as markers for the genetic characterization of A. marginale strains and phylogenetic studies. MSP1a is involved in the adhesion and transmission of A. marginale by ticks and varies among geographic strains in the number and sequence of amino-terminal tandem repeats. The aim of this study was to characterize the genetic diversity of A. marginale strains collected from countries in North and South America, Europe, Asia, Africa and Australia, inclusive of all continents. In this study, we characterized 131 strains of A. marginale using 79 MSP1a repeat sequences. These results corroborated the genetic heterogeneity of A. marginale strains in endemic regions worldwide. The phylogenetic analyses of MSP1a repeat sequences did not result in clusters according to the geographic origin of A. marginale strains but provided phylogeographic information. Seventy-eight percent of the MSP1a repeat sequences were present in strains from a single geographic region. Strong (> or =80%) support was found for clusters containing sequences from Italian, Spanish, Chinese, Argentinean and South American strains. The phylogenetic analyses of MSP1a repeat sequences suggested tick-pathogen co-evolution and provided evidence of multiple introductions of A. marginale strains from various geographic locations worldwide. These results contribute to the understanding of the genetic diversity and evolution of A. marginale and tick-pathogen interactions. PMID- 17084045 TI - Fetal testosterone and sex differences. AB - Experiments in animals leave no doubt that androgens, including testosterone, produced by the testes in fetal and/or neonatal life act on the brain to induce sex differences in neural structure and function. In this article, we argue that prenatal and neonatal testosterone exposure are strong candidates for having a causal role in sexual dimorphism in human behaviour, including social development. PMID- 17084046 TI - Endocrine, luteal and follicular responses after the use of the short-term protocol to synchronize ovulation in goats. AB - The effect of the so-called Short-Term Protocol (5-day progesterone treatment+PGF(2)alpha) on ovarian activity and LH surge was studied in goats. The goats received 250IU eCG at the time of device withdrawal (eCG group; n=7), or 200microg of EB (estradiol benzoate) 24h after device withdrawal (EB group; n=8), or received neither eCG nor EB (control group; n=8). The Short-Term Protocol induced greater (4.1+/-1.1ng/ml) progesterone serum concentrations at 24h after start of the treatment, that declined to 0.2+/-0.1ng/ml at 12h after device withdrawal. In all of the groups, the maximum concentration of estradiol-17beta was reached at about 36h after device withdrawal. Maximum concentration was greater in the EB group (76.9+/-24.6pmol/l) than in the control group (41.8+/ 9.0pmol/l; P<0.01), with the eCG group showing intermediate concentration (70.3+/ 32.5pmol/l; P=NS). The LH peak occurred earlier in the eCG group (38.4+/-2.0h after device withdrawal) and in the EB group (41.0+/-4.1h), than in the control group (46.3+/-5.1h; P<0.05). Ovulation occurred earlier in the eCG group (5/7) and in the EB group (8/8) (58.8+/-2.7h and 63.0+/-5.6h, respectively), than in the control group (7/8) (70.2+/-8.3h; P<0.05). In summary, the Short-Term Protocol induced similar concentrations of progesterone among treated goats. In addition, eCG or EB resulted in a similar increase in estradiol-17beta and a similar LH surge, which induced ovulation in most females (86.7%) in a consistent interval (about 60h) after the end of progesterone exposure. PMID- 17084047 TI - Sexual and psychological symptoms in the climacteric years. AB - OBJECTIVES: To provide epidemiological data about psychological and sexual functioning during menopausal transition in a large Italian non-clinical sample, and to investigate their correlation with life events. METHODS: The study design was a cross-sectional postal survey of a menopausal sample of women recruited from the General Registry Office in Ferrara's province. The sample was composed of four thousand and seventy-three women; they were sent a questionnaire designed on the basis of the Women's Health Questionnaire (WHQ). Together with the WHQ, the subjects filled out a personal file to define social status, cultural level, family's characteristics, recent menstrual cycles, gynaecological history and operations, drug assumption, life events in the last year, and lifetime depression. RESULTS: One thousand three hundred and forty-five women provided usable questionnaires. Factor analysis resulted in eight clusters: somatic symptoms, depressive symptoms, depressed mood with anxiety symptoms, cognitive difficulties, anxiety, sexual functioning, vasomotor symptoms and sleep problems. Mood and sexual function were impaired through the menopausal transition, with depressive and sexual symptoms being higher in the post-menopausal group compared to the pre-menopausal one. Therefore, the correlation between the two was greater in the pre- and peri-menopausal period. CONCLUSION: Depressive and sexual symptoms presented greater severity in the post-menopausal group. Both clusters of symptoms were strongly associated with life events. The parallel course of the two clusters could be related with a common pathoplastic action of life events, both on sexual symptoms and on depressive symptoms, occurring right at the time that a woman has to face the transition into menopause. PMID- 17084048 TI - Arabinogalactan protein from Arachis hypogaea: role as carrier in drug formulations. AB - Arabinogalactan protein (AGP) a highly water-soluble glyco-conjugate from groundnut (Arachis hypogaea L.) seedling was isolated and purified by precipitation with beta-glucosyl Yariv reagent. Quantification of AGP was done by gel diffusion assay. Purified AGP was conjugated to amphotericin-B (AmB) by Schiff base reaction at pH 11.0, with aim to prepare a water-injectable lesser toxic AGP-AmB conjugate without affecting AmB antifungal potential. The AGP-AmB conjugate antifungal activity was assayed by serial broth dilution and disc method against several Candida albicans clinical isolates. Both AmB and AGP-AmB showed similar MICs and MFCs activities, indicating that AGP do not reduced the antifungal activity of AmB. However, the in vitro and in vivo toxicity assays revealed that AGP-AmB conjugate was lesser toxic than AmB, as high MTD (45 mg/kg body weight) was observed. It is suggested that AGP could be a potent carrier in AmB formulation, which may result in effective treatment of fungal infections. PMID- 17084049 TI - A novel in situ forming drug delivery system for controlled parenteral drug delivery. AB - The objective of this study was to investigate the in vitro drug (diltiazem hydrochloride and buserelin acetate) release from different in situ forming biodegradable drug delivery systems, namely polymer solutions (in situ implants) and in situ microparticle (ISM) systems. The drug release from ISM systems [poly(d,l-lactide) (PLA) or poly(d,l-lactide-co-glycolide) (PLGA)-solution dispersed into an external oil phase] was investigated as a function of the type of solvent and polymer, polymer concentration and internal polymer phase:external oil phase ratio and was compared to the drug release from in situ implant systems and microparticles prepared by conventional methods (solvent evaporation or film grinding). Upon contact with the release medium, the internal polymer phase of the ISM system solidified and formed microparticles. The initial drug release from ISM systems decreased with increasing polymer concentration and decreasing polymer phase:external oil phase ratio. The type of biocompatible solvent also affected the drug release. It decreased in the rank order DMSO>NMP>2-pyrrolidone. In contrast to the release of the low molecular weight diltiazem hydrochloride, the peptide release (buserelin acetate) was strongly dependent on the polymer degradation/erosion. One advantage of the ISM system when compared to in situ implant systems was the significantly reduced burst effect because of the presence of an external oil phase. ISM systems resulted in drug release profiles comparable to the drug release of microparticles prepared by the solvent evaporation method. Therefore, the ISM systems are an attractive alternative to existing complicated microencapsulation methods. PMID- 17084050 TI - Influence of ethanol on aspirin release from hypromellose matrices. AB - Release profiles of aspirin from hypromellose matrices in hydro-ethanolic media were studied. Percent aspirin released increased with increasing levels of ethanol in the dissolution media, correlating with the drug's solubility, however, dose dumping of aspirin did not occur. An initial rapid release was observed in media comprising 40% ethanol. Release in these conditions was considered to be both erosion and diffusion-mediated, in contrast to the release in 0, 10, 20 and 30% ethanol media, where erosion-controlled release dominated. Image analysis of matrix swelling indicated a slower initial interaction between ethanol and hypromellose accounting for the initial rapid release. Cloud point studies suggested that ethanol retarded hydration of the polymer. PMID- 17084051 TI - Current perspectives in dissolution testing of conventional and novel dosage forms. AB - The purpose of this article is to review USP and non-pharmacopeial dissolution testing methods for conventional and novel pharmaceutical dosage forms and give an insight to possible alternatives in drug dissolution study design and appropriate choices for dissolution media. For each dosage form first the USP method(s) for dissolution testing are reviewed followed by alternative methods used in research and development. PMID- 17084052 TI - Olanzapine-induced hyperglycemic ketoacidosis and corresponding acetone concentrations post-mortem: a forensic interpretation. AB - Olanzapine has been shown to cause or have a contributory role in the development of hyperglycemia and diabetes mellitus. Without careful monitoring for the development of these conditions and control of the resulting adverse effects, patients receiving olanzapine may be at risk of developing fatal ketoacidosis. A review of post-mortem toxicological reports has revealed an increase in the incidence of post-mortem findings of acetone in decedents who were taking olanzapine over the past decade. A review of the current literature and a comprehensive review of case histories and toxicological findings were conducted at the Centre of Forensic Sciences (Toronto, Ontario). Olanzapine concentrations ranging from <62.5 to 858 ng/mL and acetone concentrations as high as 95 mg/dL were detected concurrently. Due to the unstable nature of olanzapine, in several instances quantitation was not possible despite elevated responses during qualitative screening procedures. Five cases suggesting olanzapine-induced ketoacidosis were identified based on the case history and toxicological findings. These data have been compiled and examined with respect to acetone concentrations following olanzapine use and the forensic relevance of post-mortem olanzapine and acetone concentrations are discussed. PMID- 17084053 TI - Multiple myeloma involving the thyroid cartilage. AB - Multiple myeloma, solitary plasmacytoma, and extramedullary plasmacytoma constitute a continuous disease spectrum of plasma cell neoplasms. In the larynx, although extramedullary plasmacytoma in the supraglottic region has been sometimes reported, plasma cell neoplasm with involvement of the thyroid cartilage is extremely rare. We report a case of multiple myeloma involving the thyroid cartilage. A 72-year-old male patient presented with a pathological fracture of the cervical vertebrae. CT scan revealed low-density areas within the thyroid cartilage, but the laryngeal mucosa and submucosal soft tissue were intact. Multiple myeloma was diagnosed, since the tumors in the thyroid cartilage and cervical vertebrae revealed plasmacytoma. Because no other lesion was found, irradiation of the larynx and cervical vertebrae was performed. Neither aggressive change of these lesions nor new lesions have been found over 3-year follow-up following the initial treatment without adjuvant therapy. PMID- 17084054 TI - Telomeres. AB - The role that telomere biology may play in the human ageing process is of a significant interest to many laboratories around the world. In this, the first of a series of yearly reviews on telomeres and ageing, I review a small selection of papers published between July 2005 and June 2006 that maybe of direct relevance to the gerontology research community. PMID- 17084055 TI - 19F NMR as a powerful technique for the assay of anti-psychotic drug haloperidol in human serum and pharmaceutical formulations. AB - 19F nuclear magnetic resonance was used as a suitable analytical tool for the identification and selective determination of haloperidol in human serum and pharmaceutical preparations. The method is based on the integration of appropriate signals of haloperidol and trifluoroacetic acid as an internal standard. The proposed method is a rapid and facile, while without any sample pretreatment, manipulation of large samples and lengthy instrument time. The regression equation for haloperidol in human serum showed a good linearity in the range of 60-600 microg ml(-1) with a detection limit of 1.4 microg ml(-1). The mean recovery results on human serum samples ranged from about 96-103%, with relative standard deviations <8%. The method was also applied successfully to the determination of haloperidol in real pharmaceutical samples, and compared with the results obtained by a reference method. The drug's degradation was studied by the proposed method in hydrochloric acid media and main products were identified. PMID- 17084056 TI - Direct quantification in bioanalytical LC-MS/MS using internal calibration via analyte/stable isotope ratio. AB - The possibility to rationalize and simplify bioanalysis, without compromising the analytical quality, by omitting the calibration curves was studied. Using mass spectrometry (MS) and a stable isotope labeled internal standard it was possible to get equally good results by calculating the results directly from the analyte/internal standard area ratio and a predetermined response factor as by the traditional way, using a calibration curve run at the same occasion. To be able to use this simplified quantification method, that we call internal calibration, in its most simple form there are some prerequisites that must be considered: (1) The relative response should not be concentration dependent. (2) The relative response should be constant between batches/days. (3) The level of analyte in the internal standard should not be detectable. (4) There should be no influence from naturally occurring isotopes of the analyte on the internal standard peak area. A bioanalytical LC-MS/MS method for a research compound was validated both with and without calibration curves and no significant differences were found regarding precision and accuracy. It was shown that all four prerequisites above were fulfilled. Validation data were very good for the whole concentration range, 0.010-30 micromol/L. Long-term data for QC samples showed excellent precision and accuracy. PMID- 17084057 TI - Simultaneous determination of betulin and betulinic acid in white birch bark using RP-HPLC. AB - A simple procedure is described for the simultaneous extraction and determination of betulin and betulinic acid in white birch bark. The extraction was checked using different solvents: dichloromethane, ethyl acetate, acetone, chloroform, methanol and 95% ethanol (aqueous solution, v/v). It was found 95% ethanol was a good extraction solvent that allowed extraction of triterpenoid with a highest content. Separation was achieved on a reversed phase C(18) column with acetonitrile-water 86:14 (v/v). Detection was accomplished with UV detection at lambda=210 nm. Using this method, the bioactive triterpenoid in white birch bark were simultaneously determined. Significant variations in the content of betulin and betulinic acid in white birch bark growing in different locations of China were also observed. PMID- 17084058 TI - A systematic review of computer-based softwares for educating patients with coronary heart disease. AB - OBJECTIVE: To evaluate the use of computer-based softwares for educating patients with coronary heart disease. METHODS: A systematic electronic search for randomised controlled trials and comparison studies published from 1999 to the end of 2005 using the MEDLINE (1999-2005), EMBASE (1999-2005) and CINAHL (1999 2005) was carried out. Articles including the reference lists in the following journals were hand-searched: Patient Education and Counselling and Patient Counselling and Health Education. RESULTS: A total of 487 articles were identified. Based on a review of abstracts, five studies fulfilled the inclusion criteria of the review. A scoring sheet was used to assess the papers' quality. All studies reported significantly increased knowledge in patients using the educational software when compared to standard education. The difference in knowledge between the intervention and control groups remained high even at 6 months follow up. Furthermore, patients reported high satisfaction with the educational programs. CONCLUSION: Despite there only being five studies that met the inclusion criteria, this review supports the successful use of computer software to increase knowledge in patients with coronary heart disease. The reviewed articles reveal that computer-based education has an important role in increasing patients' knowledge about their condition. PRACTICAL IMPLICATIONS: It is commonly reported that patients want more information about their illness. This study shows that computer-based education can be a useful, acceptable to patients and effective way to deliver education about coronary heart disease. PMID- 17084059 TI - Patient empowerment in theory and practice: polysemy or cacophony? AB - OBJECTIVE: This paper examines how the term "empowerment" has been used in relation to the care and education of patients with chronic conditions over the past decade. METHODS: Fifty-five articles were analysed, using a qualitative method of thematic analysis. RESULTS: Empowerment is more often defined according to some of its anticipated outcomes rather than to its very nature. However, because they do not respect the principle of self-determination, most anticipated outcomes and most evaluation criteria are not specific to empowerment. Concerning the process of empowerment, our analysis shows that (i) the educational objectives of an empowerment-based approach are not disease-specific, but concern the reinforcement or development of general psychosocial skills instead; (ii) empowering methods of education are necessarily patient-centred and based on experiential learning; and (iii) the provider-patient relationship needs to be continuous and self-involving on both sides. CONCLUSION: Our analysis did not allow for the unfolding of a well-articulated theory on patient empowerment but revealed a number of guiding principles and values. PRACTICE IMPLICATIONS: The goals and outcomes of patient empowerment should neither be predefined by the health-care professionals, nor restricted to some disease and treatment-related outcomes, but should be discussed and negotiated with every patient, according to his/her own particular situation and life priorities. PMID- 17084060 TI - Variations in treatment of ductal carcinoma in situ of the breast: a population based study in the East Netherlands. AB - AIMS: Differences in treatment of ductal carcinoma in situ (DCIS) of the breast were analysed for a geographically defined population in the East Netherlands. METHODS: Data from the Cancer Registry of the Comprehensive Cancer Centre East Netherlands were analysed for treatment of DCIS in the period between January 1989 and December 2003. The study population consisted of 800 female patients with a first diagnosis of DCIS of whom 798 underwent surgical treatment. The distribution of tumour characteristics and treatment were compared for several time periods. RESULTS: Surgical treatment was specified for 648 patients: 51% underwent breast-conserving surgery. The proportion of patients treated with breast-conserving surgery increased: 43% in the period 1994-1998 and 55 after 1999 (p<0.01). An axillary staging procedure was performed in 149 patients (19%), of whom 2 (1%) had tumour-involved lymph nodes. Of patients treated with breast conserving surgery, 133 (40%) received radiation therapy: 7% in the period 1994 1998 compared to 62% after 1999 (p<0.01). Patients (60%) of 50 years or younger were treated with mastectomy compared to 44% in patients aged 50-69 years and 50% in patients of 70 years and older (p<0.01). The rate in use of radiation therapy after breast-conserving surgery was comparable to both age groups. CONCLUSION: This study shows variability in the treatment of DCIS in a geographically defined region. Approximately half of all patients were treated with mastectomy and 19% underwent an axillary staging procedure; this may represent aggressive, unwarranted treatment. In contrast, 38% of patients treated with breast conserving surgery were not treated with radiation therapy after 1999, which may represent under-treatment. PMID- 17084061 TI - 1,1,2-Tris-organoselenide alkene derivatives, but not 1,2-bis-organoselenide alkene derivatives, inhibited delta-aminolevulinate dehydratase activity from human erythrocytic cells in vitro. AB - Organochalcogens are important intermediates and useful reagents in organic synthesis. Recent data from our laboratory demonstrated that bis and tris selenide alkene derivatives are attractive synthetic targets because of their chemio-, regio- and stereo-selective reactions. Since the erythrocytic delta aminolevulinate dehydratase (delta-ALA-D) activity could be an important indicator of toxicity, this report investigated bis and tris-selenide alkene derivatives effects on blood delta-ALA-D in vitro. To investigate the mechanisms by which these compounds inhibit human blood delta-ALA-D activity, a thiol reducing agent or zinc chloride were used. 1,2-Bis-selenide alkene derivatives 1a (R=4-MeOC(6)H(4)), 1b (R=4-ClC(6)H(4)) and 1c (R=2,4,6-Me(3)C(6)H(2)) did not inhibit human blood delta-ALA-D activity. 1,1,2-Tris-selenide alkene derivative 2a (R=C(6)H(5)) was the most potent delta-ALA-D inhibitor. Compounds 2b (R=4 MeOC(6)H(4)) and 2c (R=4-ClC(6)H(4)) displayed similar inhibitory potency towards delta-ALA-D activity. Dithiothreitol, a hydrophobic SH-reducing agent, was able to restore and to protect delta-ALA-D activity inhibited by tris-selenide alkene derivatives. Conversely, ZnCl(2) did not alter the enzyme inhibition induced by tris-selenide alkene derivatives. From these findings we suggest that 1,1,2-tris selenide alkene derivatives inhibited delta-ALA-D activity by an interaction with essential sulfhydryl groups for the enzyme activity. PMID- 17084062 TI - Effect of enlarged glutathione on zinc-mediated toxicity in lung-derived cell lines. AB - Zinc-mediated toxicity has been linked to cellular glutathione content in isolated cells. In addition, treatment of alveolar epithelial type II cells with glucocorticoids diminishes cellular glutathione content, and this is followed by an increase in zinc-mediated toxicity. The question arises whether an increase in glutathione synthesis might decrease zinc-mediated toxicity. For this purpose an administration of 200 micromol/l N-acetyl-L-cysteine (NAC) was given to the cells, while cysteine was used up to 100 micromol/l. Zinc-mediated toxicity was assessed by measuring protein synthesis inhibition and glutathione dependent parameters. De novo synthesis of glutathione was assessed as compared to controls by N-acetyl-D-cysteine (NADC) treatment. Comparing NAC and NADC treatment no differences in zinc-mediated toxicity were found. Furthermore only in one (of three) cell line tested a significant increase in GSH content by NAC as compared to NADC treatment was achieved. But even in this cell line no changes by zinc mediated toxicity were found. It is concluded that the cell lines tested can use other sources of cys for glutathione synthesis. Furthermore the increased zinc mediated toxicity due to hydrocortisone was abolished in the alveolar epithelial cell lines by the NADC/NAC treatment. It is therefore discussed that additionally to glutathione some other antioxidative defence mechanisms can influence zinc mediated toxicity as well. PMID- 17084063 TI - Pseudo-PTSD. AB - Pseudo-posttraumatic stress disorder (pseudo-PTSD) refers to cases in which a patient's presentation is but a simulation of the actual clinical syndrome. The problem of pseudo-PTSD has been neglected by many clinicians and researchers, who often rely on the assumption that a patient's reported symptoms can be accepted as valid. The purpose of this article is to (a) consider the diverse causes of pseudo-PTSD, (b) emphasize the importance of the DSM-IV's guideline to rule out malingering, and (c) discuss the implications that pseudo-PTSD has for research and clinical practice. PMID- 17084064 TI - Posttraumatic stress syndromes: Useful or negative heuristics? AB - The articles in this special issue provide a wide range of challenges to current conceptions, nosology, and assessment procedures for posttraumatic stress disorders. At best, they overcome the negative heuristic posed by these disorders, reopening issues that have preemptively been closed about dissociation, the presumed causal connection between a life threatening event and the symptoms of posttraumatic stress disorders, and the adequacy of checklist assessments of symptoms. They note discontinuities between current thinking about these disorders and the dominant thinking of the past. We make recommendations for more studies that similarly challenge the validity of current conceptions of posttraumatic disorders and dissociation, and the adequacy of checklist assessments of symptoms. With this goal, we note the value of studying ersatz posttraumatic stress response. Finally, we call for greater transparency in this literature with author disclosure of activity as expert witnesses. PMID- 17084065 TI - Automatic arm removal in PET and CT images for deformable registration. AB - Positron emission tomography (PET) imaging is rapidly expanding its role in clinical practice for cancer management. The high sensitivity of PET for functional abnormalities associated with cancer can be confounded by the minimal anatomical information it provides for cancer localization. Computed tomography (CT) provides detailed anatomical information but is less sensitive to pathologies than PET. Thus, combining (i.e., registering) PET and CT images would enable both accurate and sensitive cancer localization with respect to detailed patient anatomy. An additional application area of registration is to align CT-CT scans from serial studies on a patient on a PET/CT scanner to facilitate accurate assessment of therapeutic response from the co-aligned PET images. To facilitate image fusion, we are developing a deformable registration software system using mutual information and a B-spline model of the deformation. When applying deformable registration to whole body images, one of the obstacles is that the arms are present in PET images but not in CT images or are in different positions in serial CT images. This feature mismatch requires a preprocessing step to remove the arms where present and thus adds a manual step in an otherwise automatic algorithm. In this paper, we present a simple yet effective method for automatic arm removal. We demonstrate the efficiency and robustness of this algorithm on both clinical PET and CT images. By streamlining the entire registration process, we expect that the fusion technology will soon find its way into clinics, greatly benefiting cancer diagnosis, staging, therapy planning and treatment monitoring. PMID- 17084066 TI - Amelioration of water maze performance deficits by topiramate applied during pilocarpine-induced status epilepticus is negatively dose-dependent. AB - Temporal lobe epilepsy is characterized by a progressive loss of memory capacities, due to sclerosis and functional impairment of mesiotemporal brain areas. We have shown recently that topiramate (TPM) dose-dependently protects hippocampal CA1 and CA3 neurons during initial status epilepticus in the rat pilocarpine model of temporal lobe epilepsy by inhibition of mitochondrial transition pore opening. In the present study, in order to evaluate possible positive effects of the treatment on learning and memory, we investigated water maze performance of rats receiving different dosages of TPM (20 and 100 mg/kg) after 40 min and 4 mg/kg diazepam after 160 min of pilocarpine-induced status epilepticus in relation to performance of animals receiving 4 mg/kg diazepam after 40 min of SE, and to performance of sham-treated control animals. Unexpectedly, 20 but not 100 mg/kg TPM significantly extenuated short-term memory deficits. While neuroprotective effects of TPM were observed in hippocampal CA subfields of animals treated with 100 mg/kg TPM, cell loss in rats treated with 20 mg/kg TPM was indistinguishable from animals receiving diazepam only. The present results indicate a negative dose-dependency of memory-saving effects of TPM applied during status epilepticus apparently dissociated from hippocampal neuroprotection. PMID- 17084067 TI - Optimal treatment of leptospirosis: queries and projections. AB - Although the global burden of leptospirosis remains enormous and new aspects of the disease are constantly recognised, little progress has been achieved in the field of leptospirosis therapeutics and queries regarding the utility of antibiotics in the late severe form of the disease remain. From the currently existing data, conclusions on the efficacy of antibiotic administration in severe or late disease cannot easily be drawn, since clinical trials have different selection criteria and may focus on Leptospira serovars with different virulence. However, as a rule the benefit of the doubt should apply. Moreover, new options, such as ceftriaxone, have a superior safety profile to penicillin. In vitro studies have outlined potential antimicrobial candidates such as macrolides and ketolides. Development of a globally accepted subunit vaccine for humans is warranted but is not expected in the near future. PMID- 17084068 TI - Dissociating episodic from semantic access mode by mutual information measures: evidence from aging and Alzheimer's disease. AB - Re-thinking the semantic vs. episodic distinction with new experimental paradigms, we have designed a simple classification task to assess episodic and semantic access modes to memory for famous faces. The task requires to post 54 cards into nine mail boxes arranged in a 3x3 (nationality by field of activity) array, allowing for a quantitative analysis of the distribution of their responses, in particular of their classification errors. By using an information theoretical approach, we have developed an index of the concentration of errors, i.e. the metric content index. High levels of metric content indicate strong dependence of the classification performance on perceived relations among the set of stimuli, and therefore a preferred semantic access mode. We have found (1) a significant effect of age on the metric content, indicative of a shift from episodic to semantic access in older subjects (Experiment 1); (2) a significant correlation between the metric content and relevant measures assessing episodic and semantic retrieval mode in the Remember (R)/Know (K) paradigm introduced by Tulving [Tulving, E. 1985. Memory and consciousness. Can. Psychol. 26, 1-12] (Experiment 2); (3) a significant increase in metric content in early Alzheimer's disease patients compared to normal controls, consistent with their specific impairment in episodic access (Experiment 3). PMID- 17084069 TI - In vitro release of a water-soluble agent from low viscosity biodegradable, injectable oligomers. AB - Low-molecular-weight poly(epsilon-caprolactone-co-1,3-trimethylene carbonate) and poly(1,3-trimethylene carbonate) are potential vehicles for the regio-specific delivery of water-soluble agents. In this paper, the characteristics and the mechanism governing the in vitro release of a model water-soluble drug, vitamin B12, from these polymer vehicles were determined. The loading of vitamin B12 was kept to 1 w/w%. The oligomers examined ranged from amorphous, high viscosity to crystalline but low viscosity. The oligomers did not degrade appreciably in vitro. The total fraction of vitamin B12 released increased as the crystallinity of the oligomers decreased, reaching nearly total release only for the completely amorphous oligomers. The rate of release was fastest for the amorphous oligomers and dependent on their viscosity. Inclusion of a more osmotically active agent, trehalose, into the vitamin B12 particles through co-lyophilization resulted in enhanced total fraction released and a faster release rate. The results are consistent with an osmotically driven release mechanism. PMID- 17084070 TI - Use of selected waste materials in concrete mixes. AB - A modern lifestyle, alongside the advancement of technology has led to an increase in the amount and type of waste being generated, leading to a waste disposal crisis. This study tackles the problem of the waste that is generated from construction fields, such as demolished concrete, glass, and plastic. In order to dispose of or at least reduce the accumulation of certain kinds of waste, it has been suggested to reuse some of these waste materials to substitute a percentage of the primary materials used in the ordinary portland cement concrete (OPC). The waste materials considered to be recycled in this study consist of glass, plastics, and demolished concrete. Such recycling not only helps conserve natural resources, but also helps solve a growing waste disposal crisis. Ground plastics and glass were used to replace up to 20% of fine aggregates in concrete mixes, while crushed concrete was used to replace up to 20% of coarse aggregates. To evaluate these replacements on the properties of the OPC mixes, a number of laboratory tests were carried out. These tests included workability, unit weight, compressive strength, flexural strength, and indirect tensile strength (splitting). The main findings of this investigation revealed that the three types of waste materials could be reused successfully as partial substitutes for sand or coarse aggregates in concrete mixtures. PMID- 17084071 TI - DNA microdevice for electrochemical detection of Escherichia coli 0157:H7 molecular markers. AB - An electrochemical DNA sensor based on the hybridization recognition of a single stranded DNA (ssDNA) probe immobilized onto a gold electrode to its complementary ssDNA is presented. The DNA probe is bound on gold surface electrode by using self-assembled monolayer (SAM) technology. An optimized mixed SAM with a blocking molecule preventing the nonspecific adsorption on the electrode surface has been prepared. In this paper, a DNA biosensor is designed by means of the immobilization of a single stranded DNA probe on an electrochemical transducer surface to recognize specifically Escherichia coli (E. coli) 0157:H7 complementary target DNA sequence via cyclic voltammetry experiments. The 21 mer DNA probe including a C6 alkanethiol group at the 5' phosphate end has been synthesized to form the SAM onto the gold surface through the gold sulfur bond. The goal of this paper has been to design, characterise and optimise an electrochemical DNA sensor. In order to investigate the oligonucleotide probe immobilization and the hybridization detection, experiments with different concentration of DNA and mismatch sequences have been performed. This microdevice has demonstrated the suitability of oligonucleotide Self-assembled monolayers (SAMs) on gold as immobilization method. The DNA probes deposited on gold surface have been functional and able to detect changes in bases sequence in a 21-mer oligonucleotide. PMID- 17084072 TI - An integrated and sensitive detection platform for magneto-resistive biosensors. AB - A compact biosensor platform with giant magneto-resistive (GMR) sensors suited for the detection of superparamagnetic nanoparticle labels is presented. The platform consist of disposable biosensor cartridges and an electronic reader, which enables quantitative detection with high analytical performance, combined with robustness, ease of use and at low cost. In order to optimise the signal-to noise ratio (SNR), magnetic labels are excited at high frequency. Wires, integrated in the silicon of the sensor chip are used to generate a well-defined magnetic field on the sensor surface, thus removing the need for mechanical alignment with external apparatus. A signal modulation scheme is applied to obtain optimal detection accuracy. The platform is scalable and can be adapted according to application-specific requirements. Experimental results indicate that three beads of 300 nm diameter can be detected on a sensor surface of 1500 microm2 for a measurement time of 1s. PMID- 17084073 TI - Lining the pockets of kinases and phosphatases. AB - The regulation of the activity of kinases and phosphatases is an essential aspect of intracellular signal transduction. Recently determined structures of AGC protein kinases, including isoforms of PKB, PKC, GRK and ROCK, indicate that occupancy of a hydrophobic pocket in the kinase N-lobe by a segment of the protein immediately C terminal to the kinase domain provides a mechanism for regulating kinase activity. In addition, crystal structures of Aurora-A and Aurora-B, which are closely related to AGC family kinases, in complex with their activators, TPX2 and INCENP, respectively, show how allosteric kinase activation is achieved by the binding of the activator protein to an equivalent hydrophobic pocket. Hence, regulation of kinase activity by analogous interactions is a shared regulatory mechanism of these kinases. Two crystal structures have explained the molecular basis of PKA anchoring through its regulatory subunits by members of the AKAP family of scaffold proteins. AKAPs can also interact directly with protein kinase and phosphatase catalytic domains. The crystal structure of the PP1 catalytic subunit in complex with the targeting subunit MYPT1 indicates that there is also scope for intimate phosphatase regulation by scaffold proteins. PMID- 17084074 TI - Targeting ubiquitin in cancers. AB - Ubiquitin (Ub) is a small protein modifier involved in cellular functions such as cell cycle, apoptosis, cell signalling, endocytosis, transcription and DNA repair. Ubiquitin operates as a reversible and highly versatile regulatory signal, which may be read and interpreted by an expanding number of Ub-binding domains (UBD). There is accumulating evidence that mutations or altered expression of ubiquitylating or de-ubiquitylating enzymes as well as of Ub binding proteins affect crucial mediators of such functions and are found in several malignancies. Here we discuss how oncogenic alterations in the Ub system can be targeted by anti-cancer therapies. PMID- 17084075 TI - Progression of localised Wilms' tumour during preoperative chemotherapy is an independent prognostic factor: a report from the SIOP 93-01 nephroblastoma trial and study. AB - The SIOP nephroblastoma clinical trials have previously demonstrated that preoperative chemotherapy is advantageous for patients with nephroblastoma (Wilms' tumour). However, some primary tumours increase in size during preoperative chemotherapy, and to investigate the clinical relevance of this progression we studied the patient cohort with increasing tumours included in the SIOP 93-01 study (June 1993 to June 2000). Patients were considered eligible if they had a confirmed localised Wilms' tumour that had been measured in at least two dimensions at diagnosis and before surgery. Tumour response to preoperative chemotherapy was defined according to criteria set by the World Health Organisation (WHO). Patient characteristics in the different response groups were compared and related to event-free survival and overall survival. Patient records were studied regarding compliance with protocol. Tumour progression during preoperative chemotherapy was observed in 57 of 1090 patients (5%) with localised Wilms' tumours. In those cases, the tumours were significantly smaller at diagnosis and were more often stage III (p=0.05) and associated with high risk histopathology (p=0.03). After adjustment for stage and risk group, progression was proved to be correlated with poorer event-free and overall survival (hazard ratio 1.9, p=0.026 and 3.2, p=0.002 respectively). In summary, progression of localised Wilms' tumours is rarely seen in patients during preoperative chemotherapy. However, independent of stage distribution and histopathological risk group, those whose tumours do increase in size have poorer event-free and overall survival. PMID- 17084076 TI - Effects of testosterone on hormonal content and calcium-dependent basal secretion in female rat pituitary cells. AB - In vivo and in vitro effects of elevated androgens on agonist-induced gonadotropin secretion have been addressed previously. Here we investigated the effects of testosterone on hormonal content and basal (in the absence of agonists) hormone release in pituitary lactotrophs, somatotrophs and gonadotrophs from female rats. Furthermore we tested the hypothesis that testosterone action is dependent on the pattern of spontaneous and Bay K 8644 (a L-type calcium channel agonist) -induced calcium signalling. Mixed anterior pituitary cells were cultured in steroid containing or depleted media, and testosterone (1pM to 10nM) was added for 48h. Cells were studied for their spontaneous and Bay K 8644 induced calcium signalling pattern and total hormone levels (release and hormonal content). In lactotrophs, somatotrophs and gonadotrophs testosterone did not affect the pattern of spontaneous calcium signalling. Bay K 8644-induced calcium signalling and hormone release were not affected by testosterone. In both steroid depleted and -containing medium, testosterone inhibited prolactin (PRL), luteinizing hormone (LH) and growth hormone (GH) cellular content and release in a dose-dependent manner, with IC(50)s in a sub-nanomolar concentration range. These results indicate that testosterone inhibits basal hormone release from lactotrophs, somatotrophs and gonadotrophs without affecting intracellular calcium signalling. This action of testosterone is not dependent on the presence of other steroid hormones. PMID- 17084077 TI - Removal of ammonia as struvite from anaerobic digester effluents and recycling of magnesium and phosphate. AB - A second order kinetic model was developed to predict the rate and extent of NH(4)(+) removal as struvite from anaerobic digester effluents. Alternative to this, NH(4)(+) can be recovered from struvite and the remaining Mg(2+) and PO(4)(3-) can be recycled back to the wastewater to fix more NH(4)(+). The NH(4)(+) solution was retained and the remaining Mg(2+) and PO(4)(3-) were returned back to be mixed with wastewater. In a five-step process, NH(4)(+) recovery was initially 92% and progressively decreased to 77% in the fifth stage, due to loss of Mg(2+) and PO(4)(3-) at each step in the supernatant. Finally, economic analysis of recycling nutrients was performed and compared to the one step process. The cost of NH(4)(+) recovery was calculated as $0.36/kgNH(4)-N which is lower than $7.7/kgNH(4)-N the cost of one step process without considering the market value of struvite obtained in one step process. PMID- 17084078 TI - Removal of Astrazon Yellow 7GL from aqueous solutions by adsorption onto wheat bran. AB - Adsorption kinetic and equilibrium of a basic dye (Astrazon Yellow 7GL) from aqueous solutions at various initial dye concentration (50-300 mg/l), pH (4-10), adsorbent dosage (2-8 g/l), particle size (354-846 microm) and temperature (30-50 degrees C) on wheat bran were studied in a batch mode operation. The result showed that the amount adsorbed of the dye increased with increasing initial dye concentration and contact time, whereas particle size and pH had no significant affect on the amount of dye adsorbed by the adsorbent. A comparison of kinetic models on the overall adsorption rate showed that dye/adsorbent system was best described by the pseudo second-order rate model. The removal rate was also dependent on both external mass transfer and intra-particle diffusion. The low value of the intraparticle diffusivity, 10(-11) cm2/s, indicated the significant influence of intraparticle diffusion on the kinetic control. The adsorption capacity (Q0) calculated from the Langmuir isotherm was 69.06 mg/g for at pH 5.6, 303 K for the particle size of 354 microm. The experimental data yielded excellent fits with Langmuir and Tempkin isotherm equations. Different thermodynamic parameters showed that the reaction was spontaneous and endothermic in nature. PMID- 17084079 TI - Starch industry wastewater as a substrate for antagonist, Trichoderma viride production. AB - Starch industry wastewater was investigated to assess and improve its potential as a raw material for the conidia production of biocontrol fungi, Trichoderma viride. The wastewater was tested with and without supplements of glucose, soluble starch, meat peptone and probable conidiation inducer chemicals in shake flask culture. Addition of complex carbon source (soluble starch, 1% and 2% w/v) produced maximum conidia ( approximately 3.02 and 4.2 x 10(10)CFU/mL, respectively). On the other hand, glucose addition as a simpler carbon source was either ineffective or, reduced conidia production (from 1.6 x 10(8) in control to 3.0 x 10(7)CFU/mL in 5% w/v glucose supplement). Supplement of nitrogen source showed a small increase of conidia concentration. Propionic, maleic and humic acids, EDTA, pyridine, glycerol and CaCO(3) were examined as probable conidiation inducers and showed effect only on initial rate of conidiation with no increase in final conidia concentration. Intra and extracellular ATP correlation with spore production showed dependence on growth media used and conidia concentration at the end of fermentation. Addition of carbon and nitrogen sources showed an increase in protease activity (from 0.4985 to 2.43 IU/mL) and entomotoxicity (from 10448 to 12335 spruce budworm unit (SBU)/microL). Entomotoxicity was improved by 11% in fermenter over shake flask when starch industry wastewater was supplemented with meat peptone. PMID- 17084080 TI - Studies towards the identification of a new generation of atypical antipsychotic agents. AB - A rational structure-activity relationship study around compound (1) is reported. The lead optimisation programme led to the identification of sulfonamide (25), a molecule combining dopamine D2/D3 receptor antagonism with serotonin 5-HT2A, 5 HT2C, 5-HT6 receptor antagonism for an effective treatment of schizophrenia. Compound (25) was shown to possess the required in vivo activity with no EPS liability. PMID- 17084081 TI - Antiviral 2,5-disubstituted imidazo[4,5-c]pyridines: from anti-pestivirus to anti hepatitis C virus activity. AB - A novel class of inhibitors of the hepatitis C virus [substituted 2-(2 fluorophenyl)-5H-imidazo[4,5-c]pyridines] is described. Introduction of a fluorine in position 2 of the 2-phenyl substituent of the lead anti-pestivirus compound 1 (5-[(4-bromophenyl)methyl]-2-phenyl-5H-imidazo[4,5-c]pyridine) resulted in an analogue with selective activity against HCV in the subgenomic replicon system. PMID- 17084082 TI - The discovery of 6-[2-(5-chloro-2-{[(2,4-difluorophenyl)methyl]oxy}phenyl)-1 cyclopenten-1-yl]-2-pyridinecarboxylic acid, GW848687X, a potent and selective prostaglandin EP1 receptor antagonist for the treatment of inflammatory pain. AB - The discovery of a series of selective EP1 receptor antagonists based on a 1,2 diarylcyclopentene template is described. After defining the structural requirements for EP1 potency and selectivity, heterocyclic rings were incorporated to reduce logD and improve in vitro pharmacokinetic properties. The 2,6-substituted pyridines and pyridazines gave an appropriate balance of potency, in vivo pharmacokinetic properties and a low potential for inhibiting a range of CYP450 enzymes. From this series, GW848687X was shown to have an excellent profile in models of inflammatory pain and was selected as a development candidate. PMID- 17084083 TI - Towards erythropoietin mimicking small molecules. AB - Small molecules potentially mimicking the hormone erythropoietin have been discovered by screening of a library of rationally designed multicomponent reaction molecules in a functional cell-based assay. PMID- 17084084 TI - Evaluation of a paediatric early warning tool--claims unsubstantiated. PMID- 17084086 TI - Unified QSAR approach to antimicrobials. Part 2: predicting activity against more than 90 different species in order to halt antibacterial resistance. AB - There are many different kinds of pathogenic bacteria species with very different susceptibility profiles to different antibacterial drugs. One limitation of QSAR models is that they consider the biological activity of drugs against only one species of bacteria. In a previous paper, we developed a unified Markov model to describe the biological activity of different drugs tested in the literature against some antimicrobial species. Consequently, predicting the probability with which a drug is active against different species of bacteria with a single unified model is a goal of major importance. The work described here develops the unified Markov model to describe the biological activity of more than 70 drugs from the literature tested against 96 species of bacteria. We applied linear discriminant analysis (LDA) to classify drugs as active or inactive against the different tested bacterial species. The model correctly classified 199 out of 237 active compounds (83.9%) and 168 out of 200 inactive compounds (84%). Overall training predictability was 84% (367 out of 437 cases). Validation of the model was carried out using an external predicting series, with the model classifying 202 out of 243 (i.e., 83.13%) of the compounds. In order to show how the model functions in practice, a virtual screening was carried out and the model recognized as active 84.5% (480 out of 568) antibacterial compounds not used in the training or predicting series. The current study is an attempt to calculate within a unified framework the probabilities of antibacterial action of drugs against many different species. PMID- 17084085 TI - Epac proteins: multi-purpose cAMP targets. AB - Epac1 and Epac2 are cAMP-dependent guanine-nucleotide-exchange factors for the small GTPases Rap1 and Rap2, and are known to be important mediators of cAMP signaling. The recent determination of the crystal structure of Epac2 has indicated a mechanism for the activation of the multi-domain Epac proteins. In addition, these proteins have been implicated in various cellular processes such as integrin-mediated cell adhesion and cell-cell junction formation, the control of insulin secretion and neurotransmitter release. In most of these processes, cAMP signaling through protein kinase A (PKA) is also involved, stressing the interconnectivity between Epac- and PKA-mediated signaling. PMID- 17084087 TI - Design and synthesis of rho kinase inhibitors (III). AB - The structure-activity relationship of Rho kinase inhibitors bearing an isoquinoline scaffold was studied. N-(1-Benzyl-3-pyrrolidyl)-N-(5 isoquinolyl)amine analogues were optimized with respect to their inhibitory potencies for the enzyme and for chemotaxis. The potent analogues were further evaluated by an ex vivo test in which the selected compounds were orally administered to rats, and the Rho kinase inhibitory potency observed in the rat serum was evaluated 3h after the administration. Compound 23g showed a high level of Rho kinase inhibitory activity in the rat serum and was stable in an in vitro metabolic test using a microsomal cytochrome preparation. The (R)-isomer of 23g displayed a higher level of inhibitory potency than the (S)-isomer in a cell-free kinase assay and in the cell migration assay (IC(50)(ENZ)=25 nM and IC(50)(MCP)=1 microM). The (R)-isomer successfully inhibited the phosphorylation of MBS (myosin binding subunit) in cells. PMID- 17084088 TI - Destination of titanium particles detached from titanium plasma sprayed implants. AB - Small titanium particles may detach from titanium plasma sprayed (TPS) implants during implant insertion, when no preliminary tapping is used, probably for the frictional force between titanium coating and host bone. Aim of this study was to investigate the destination of these titanium particles observed in the peri implant environment. Twenty-four TPS screws were implanted in tibiae of two sheep. Fourteen and 90 days after implantation the implants with the surrounding bone were removed and processed to be analyzed by light microscope and scanning electron microscope (secondary electron and back-scattered electron probes). Small titanium particles detached from the unloaded TPS implants were observed both in the newly-formed bone matrix and in marrow tissue. Histomorphometric analysis showed that both at 14 and 90 days after implantation the titanium particles appeared more concentrated in marrow tissue than in calcified bone matrix, decreasing by 66.4% over time. In particular, smaller particles (<250 microm(2)) decreased by 81.5%, whereas the larger ones (250-2000 microm(2)) did not show any significant variations over time, suggesting that most of the smaller particles may undergo to ionic dissolution, probably migrating into the peri-implant marrow lacunae. A slight migration of titanium particles from the implant surface towards the more distant peri-implant tissues was also demonstrated over time. PMID- 17084089 TI - Linkage and branch determination of N-linked oligosaccharides using sequential degradation/closed-ring chromophore labeling/negative ion trap mass spectrometry. AB - A method based on sequential degradation, p-aminobenzoic ethyl ester (ABEE) closed-ring labeling, and negative ion electrospray ionization tandem mass spectrometry is presented for the study of linkage and branch determination for N linked oligosaccharides. Closed-ring labeling provides greater linkage information than the more popular open-ring reductive amination approach. In addition, after high-performance liquid chromatography (HPLC) separation, closed ring labeling allows for regeneration of the underivatized oligosaccharide, a requirement for alkaline sequential degradation. The analytical scheme presented here uses HPLC separation of closed-ring labeled oligosaccharides to resolve the mixture into individual forms that undergo subsequent structural analysis by negative ion tandem mass spectrometry. To facilitate complete structural analysis, particularly for larger sugars, the closed-ring labels are removed and the sugars are sequentially degraded by controlled alkaline hydrolysis. It is noteworthy that for sugars containing sialic acid moieties, a protecting group must be used to stabilize sialic acid groups during sequential alkaline degradation. This described approach was applied to two high mannose oligosaccharides M5G2, M6G2 cleaved from the ribonuclease B and a complex oligosaccharide A2 cleaved from transferrin. PMID- 17084090 TI - Analysis of tear glucose concentration with electrospray ionization mass spectrometry. AB - We have developed a mass spectrometry-based method that allows one to accurately determine the glucose concentration of tear fluid. We used a 1 microL micro capillary to collect tear fluid from the tear meniscus with minimal irritation of the eye. We analyzed the 1 muL volume of collected tear fluid with liquid chromatography electrospray ionization mass spectrometry with the use of D glucose-6,6-d2 as an internal standard. Repeated measurements and a recovery experiment on pooled, onion-induced tears showed that the analysis of the glucose in tears was precise (4% relative standard deviation) and provided 100% recovery. We found the tear glucose concentration of one fasting nondiabetic subject to be 13 to 51 microM while the onion-induced tear glucose concentration of a different nondiabetic subject to be 211 to 256 microM. PMID- 17084091 TI - Resolution and structural transitions of elongated states of ubiquitin. AB - Electrospray ionization, combined with two-dimensional ion mobility spectrometry and mass spectrometry, is used to produce, select, and activate distributions of elongated ions, [M + 11H]11+ to [M + 13H]13+, of ubiquitin. The analysis makes it possible to examine state-to-state transitions for structural types, and transition diagrams associated with the efficiencies of structural changes are presented. The +11 and +12 charge states can form four resolvable states while only one state is formed for [M + 13H]13+. Some conformations, which appear to belong to the same family based on mobility analysis of different charge states, undergo similar transitions, others do not. Activation of ions that exist in low abundance conformations, having mobilities that fall in between sharp peaks associated with higher abundances species, shows that the low-abundance forms undergo efficient (approximately 90 to 100%) conversion into states associated with well-defined peaks. This efficiency is significantly higher than the approximately 10 to 60% efficiency of transitions of structures associated with well-defined peaks. The formation of sharp features from a range of low-intensity species with different cross sections indicates that large regions of conformation space must be unfavorable or inaccessible in the gas phase. These results are compared with several previous IMS measurements of this system as well as information about gas-phase structure provided by other techniques. PMID- 17084092 TI - Optimization of 25 kDa linear polyethylenimine for efficient gene delivery. AB - A 25-kDa linear polyethylenimine (25 kDa L-PEI) has proven to be efficient and versatile agent for gene delivery. Therefore, we determined the optimal transfection conditions of 25 kDa L-PEI and examined whether it has comparable transfection efficiency with other commercially available reagents, ExGen 500, LipofectAMINE 2000, and Effectene by using EGFP expression vector in different cell lines. Transfection efficiency and cytotoxicity were measured by flow cytometry. First of all, we determined the optimal ratio of nitrogen to phosphorous (N/P) and DNA concentration. With the increase of N/P ratio and DNA amounts, transfection efficiency increased with a slight variation in cell types. The optimal amounts of 25 kDa L-PEI were determined at N/P ratio 40 and DNA concentration varied among the cell types. In addition, 25 kDa L-PEI worked efficiently and was less toxic than other reagents. However, the efficiency and toxicity of all these reagents varied according to cell types as well as the ratio of DNA to reagents and the amounts of DNA. Our finding illustrates the importance of optimal transfection conditions of 25 kDa L-PEI to obtain maximal transgene expression with less cytotoxicity. Importantly, the optimization of those conditions may make possible to perform transfection cost-effectively and efficiently. PMID- 17084093 TI - A versatile system for site-specific enzymatic biotinylation and regulated expression of proteins in cultured mammalian cells. AB - We have developed a system for producing biotinylated recombinant proteins in mammalian cells. The expression construct consists of an inducible tetracycline response element (TRE) that drives expression of a bicistronic cassette comprising a biotin acceptor peptide (BioTag) fused to either terminus of the target protein, the gene for Escherichia coli biotin ligase (BirA), and an intervening internal ribosome entry site (IRES). By either transient or stable transfection of Chinese hamster ovary (CHO) Tet-On cells, we successfully expressed, detected, and immobilized biotinylated human Itch, a pleiotropic multi domain ubiquitin-protein ligase, as well as Gla-RTK, a putative vitamin K dependent receptor tyrosine kinase. The biotinylation of recombinant Itch in transiently transfected CHO Tet-On cells required biotin supplementation and coexpression of BirA, occurred quantitatively and specifically on the lysine residue of the BioTag, and enabled detection of Itch by Western blot in as little as 10ng of total lysate protein. Stably selected clones were rapidly pre-screened for doxycycline (dox)-inducible BirA expression by ELISA, and subsequently screened for dox-inducible expression of biotinylated Itch. Biotinylated Gla-RTK was detectable in as little as 5ng of total lysate protein from transiently transfected CHO Tet-On cells, and exhibited pronounced tyrosine phosphorylation. In stable clones however, constitutive phosphorylation was prevented by reducing the expression level of Gla-RTK through the titration of dox. These results demonstrate the utility of this system for the expression of 'difficult' proteins, particularly those that are cytotoxic or those that may require lower expression levels to ensure appropriate post-translational modification. PMID- 17084094 TI - The identification of catalytic pentad in the haloalkane dehalogenase DhmA from Mycobacterium avium N85: reaction mechanism and molecular evolution. AB - Haloalkane dehalogenase DhmA from Mycobacterium avium N85 showed poor expression and low stability when produced in Escherichia coli. Here, we present expression DhmA in newly constructed pK4RP rhodococcal expression system in a soluble and stable form. Site-directed mutagenesis was used for the identification of a catalytic pentad, which makes up the reaction machinery of all currently known haloalkane dehalogenases. The putative catalytic triad Asp123, His279, Asp250 and the first halide-stabilizing residue Trp124 were deduced from sequence comparisons. The second stabilizing residue Trp164 was predicted from a homology model. Five point mutants in the catalytic pentad were constructed, tested for activity and were found inactive. A two-step reaction mechanism was proposed for DhmA. Evolution of different types of catalytic pentads and molecular adaptation towards the synthetic substrate 1,2-dichloroethane within the protein family is discussed. PMID- 17084095 TI - Comparative evaluation of positive tests to Mycobacterium avium subsp. paratuberculosis in clinically healthy sheep and goats in south-west Greece using molecular techniques, serology, and culture. AB - Mycobacterium avium subsp. paratuberculosis (MAP) is the cause of paratuberculosis, which affects mainly ruminants although there is a growing concern about its possible implication in Crohn's disease in humans especially in connection with environmental spread and risks to the food chain. Retail cheese may represent a significant source of human exposure to MAP and the aim of this study was to assess MAP status in clinically healthy sheep and goats in Greece, comparing techniques routinely used in the positive diagnosis of the disease. From a total of 30 flocks, 632 sheep and goats had faecal, serum, and whole-blood samples examined by culture, complement fixation test (CFT), and polymerase chain reaction (PCR) targeted at IS900, IS1245, and IS6110. PCR produced positive results in 21% of the animals tested, with 5.6%, 3.9%, and 11.5% being identified as MAP, Mycobacterium avium subsp. avium, and Mycobacterium tuberculosis complex, respectively. CFT produced positive and suspicious results in 4.4% and 14.4% of the cases. Faecal cultures were negative in all but a single case that was identified as restriction fragment length polymorphism (RFLP)-type BC1. Agreement between results obtained by PCR and CFT was poor with isolated cases although an assessment of the MAP positive tests produced similar results for both methods. The findings indicate the need for additional measures of control, although the costs may be substantial if public health protection justifies elimination of MAP from livestock. PMID- 17084096 TI - Sensitivity and resolution enhancement in solid-state NMR spectroscopy of bicelles. AB - Magnetically aligned bicelles are becoming attractive model membranes to investigate the structure, dynamics, geometry, and interaction of membrane associated peptides and proteins using solution- and solid-state NMR experiments. Recent studies have shown that bicelles are more suitable than mechanically aligned bilayers for multidimensional solid-state NMR experiments. In this work, we describe experimental aspects of the natural abundance (13)C and (14)N NMR spectroscopy of DMPC/DHPC bicelles. In particular, approaches to enhance the sensitivity and resolution and to quantify radio-frequency heating effects are presented. Sensitivity of (13)C detection using single pulse excitation, conventional cross-polarization (CP), ramp-CP, and NOE techniques are compared. Our results suggest that the proton decoupling efficiency of the FLOPSY pulse sequence is better than that of continuous wave decoupling, TPPM, SPINAL, and WALTZ sequences. A simple method of monitoring the water proton chemical shift is demonstrated for the measurement of sample temperature and calibration of the radio-frequency-induced heating in the sample. The possibility of using (14)N experiments on bicelles is also discussed. PMID- 17084097 TI - Practical aspects of (1)H transverse paramagnetic relaxation enhancement measurements on macromolecules. AB - The use of (1)H transverse paramagnetic relaxation enhancement (PRE) has seen a resurgence in recent years as method for providing long-range distance information for structural studies and as a probe of large amplitude motions and lowly populated transient intermediates in macromolecular association. In this paper we discuss various practical aspects pertaining to accurate measurement of PRE (1)H transverse relaxation rates (Gamma(2)). We first show that accurate Gamma(2) rates can be obtained from a two time-point measurement without requiring any fitting procedures or complicated error estimations, and no additional accuracy is achieved from multiple time-point measurements recorded in the same experiment time. Optimal setting of the two time-points that minimize experimental errors is also discussed. Next we show that the simplistic single time-point measurement that has been commonly used in the literature, can substantially underestimate the true value of Gamma(2), unless a relatively long repetition delay is employed. We then examine the field dependence of Gamma(2), and show that Gamma(2) exhibits only a very weak field dependence at high magnetic fields typically employed in macromolecular studies. The theoretical basis for this observation is discussed. Finally, we investigate the impact of contamination of the paramagnetic sample by trace amounts (5%) of the corresponding diamagnetic species on the accuracy of Gamma(2) measurements. Errors in Gamma(2) introduced by such diamagnetic contamination are potentially sizeable, but can be significantly reduced by using a relatively short time interval for the two time-point Gamma(2) measurement. PMID- 17084098 TI - Optimization of electrostatics as a strategy for cold-adaptation: a case study of cold- and warm-active elastases. AB - Adaptation to both high and low temperatures requires proteins with special properties. While organisms living at or close to the boiling point of water need to have proteins with increased stability, other properties are required at temperatures close to the freezing point of water. Indeed, it has been shown that enzymes adapted to cold environments are less resistant to heat with a concomitant increased activity as compared to their warm-active counter-parts. Several recent studies have pointed in the direction that electrostatic interactions play a central role in temperature adaptation, and in this study we investigate the role such interactions have in adaptation of elastase from Atlantic salmon and pig. Molecular dynamics (MD) simulations have been used to generate structural ensembles at 283 and 310 K of the psychrophilic and mesophilic elastase, and a total of eight 12 ns simulations have been carried out. Even though the two homologues have a highly similar three-dimensional structure, the location and number of charged amino acids are very different. Based on the simulated structures we find that very few salt-bridges are stable throughout the simulations, and provide little stabilization/destabilization of the proteins as judged by continuum electrostatic calculations. However, the mesophilic elastase is characterized by a greater number of salt-bridges as well as a putative salt-bridge network close to the catalytic site, indicating a higher rigidity of the components involved in the catalytic cycle. In addition, subtle differences are also found in the electrostatic potentials in the vicinity of the catalytic residues, which may explain the increased catalytic efficiency of the cold-adapted elastase. PMID- 17084099 TI - Metabolic responses and arginine kinase expression under hypoxic stress of the kuruma prawn Marsupenaeus japonicus. AB - In response to hypoxia at PO(2) 1.3-1.7 mg/L for 6 h, the kuruma prawn Marsupenaeus (Penaeus) japonicus showed a dramatic decrease in phosphoarginine storage in muscle, with normal levels restored during 4-h post-hypoxic recovery. Large stores of muscle glycogen only decreased between 4 and 6 h during hypoxia, but greatly diminished during recovery. Muscle ATP levels and energy charge decreased only slightly under hypoxia. Lactate levels increased slightly during hypoxia and promptly returned to control levels during recovery. These data indicate that phosphoarginine works in muscle as an ATP buffer during hypoxia and glycogen is utilized as an energy source during recovery. Under hypoxia, up- and down-regulated proteins were identified after 2D electrophoresis and partial sequences were obtained after protease digestion. Fructose bisphosphate aldolase was down-regulated during hypoxia, suggesting the suppression of glycolysis under hypoxia. Several partial sequences from three protein spots up-regulated under hypoxia were all assigned to arginine kinase, suggesting the existence of several isoforms of arginine kinase in the muscle of M. japonicus. This arginine kinase up-regulation under hypoxia may indicate a provision for oxygen re-supply after anaerobiosis. This is consistent with the prompt replenishment of phosphoarginine stores during recovery from hypoxia. PMID- 17084100 TI - Role of growth hormone and insulin-like growth factor-1 in the protective effect of ghrelin in ischemia/reperfusion-induced acute pancreatitis. AB - Ghrelin, an endogenous ligand for the growth hormone secretagogue receptor, has been shown to exhibit gastroprotective properties. The aim of present study was to determine whether ghrelin administration protects the pancreas against ischemia/reperfusion-induced pancreatitis and, if so, what is the role of growth hormone (GH) and insulin-like growth factor-1 (IGF-1) in this effect. In sham operated or hypophysectomized rats, acute pancreatitis was induced by pancreatic ischemia followed by reperfusion. Ghrelin (4, 8 or 16 nmol/kg/dose) or IGF-1 (20 nmol/kg/dose) were administered intraperitoneally twice before and during induction of acute pancreatitis. In pituitary-intact rats, treatment with ghrelin attenuated the development of ischemia/reperfusion-induced pancreatitis and this effect was associated with partial reversion of the pancreatitis-evoked decrease in serum concentration of GH and IGF-1. Hypophysectomy eliminated GH from the serum, reduced serum IGF-1 concentration by 90% and increased in the severity of ischemia/reperfusion-induced pancreatitis. Administration of ghrelin was without any beneficial effect in this group of rats. In contrast, administration of IGF-1 in hypophysectomized rats reduced the severity of ischemia/reperfusion-induced pancreatitis in hypophysectomized rats. We conclude that administration of ghrelin inhibits the development of ischemia/reperfusion-induced pancreatitis and this effect is mediated by its influence on the release of GH and IGF-1. PMID- 17084101 TI - Formyl peptide receptors: a promiscuous subfamily of G protein-coupled receptors controlling immune responses. AB - The formyl peptide receptor (FPR) family is involved in host defence against pathogens, but also in sensing internal molecules that may constitute signals of cellular dysfunction. It includes three subtypes in human and other primates. FPR responds to formyl peptides derived from bacterial and mitochondrial proteins. FPRL1 displays a large array of exogenous and endogenous ligands, including the chemokine variant sCKbeta8-1, the neuroprotective peptide humanin, and lipoxin A4. Two high affinity agonists (F2L and humanin) were recently described for FPRL2. In mouse, eight FPR-related receptors have been described. Fpr1 is the ortholog of human FPR, while fpr2 appears to share many ligands with human FPRL1. Altogether, the physiological role of the FPR family is still incompletely understood, due in part to the large variety of ligands, the redundancy with other chemoattractant agents, and the lack of clear orthologs between human and mouse receptors. Newly developed tools will allow to study further this family of receptors. PMID- 17084102 TI - Clinical and laboratory testing of a silver-impregnated lens case. AB - PURPOSE: Lens case contamination is a well-documented occurrence for contact lens wearers despite the efficacy of current lens disinfectants. Several microorganisms have a propensity to attach to surfaces and may become more tolerant of disinfection upon attachment. Non-compliance with disinfection regimens occasionally occurs wherein patients store their lenses in saline or in tap water. A silver-impregnated lens case was developed in an attempt to decrease case contamination. These studies examine the performance of the case both in vitro and in vivo. METHODS: In vitro, lens cases were challenged with 10(3) microorganism suspensions of bacteria. After 24h incubation at room temperature, aliquots of inocula were removed and spread plated onto appropriate growth media. Surviving colonies were counted and microorganism log drop values from initial challenge inocula were determined. Two 40-subject 1-month contralateral clinical studies were conducted with SOLO-care Aqua solution using one bowl of a silver impregnated case and one bowl of a standard lens case for lens storage. Lens cases were collected and cultured for aerobic bacteria. RESULTS: In vitro efficacy data show significantly lower numbers of recovered microbes from silver impregnated cases than from control cases. In both clinical studies, silver impregnated cases had a statistically significantly lower proportion of bacterial contamination than control cases. The majority of microorganisms isolated from silver-impregnated cases were members of the normal skin flora. CONCLUSION: The performance of the silver-impregnated case in vitro and the observed lower proportion of contaminated silver-impregnated lens cases in a clinical setting demonstrate the case's ability to decrease bacterial contamination. PMID- 17084104 TI - Studies on some salicylaldehyde Schiff base derivatives and their complexes with Cr(III), Mn(II), Fe(III), Ni(II) and Cu(II). AB - The formation constants of some transition metal ions Cr(III), Mn(II), Fe(III), Ni(II) and Cu(II) binary complexes containing Schiff bases resulting from condensation of salicylaldehyde with aniline (I), 2-aminopyridine (II), 4 aminopyridine (III) and 2-aminopyrimidine (IV) were determined pH-metrically in ethanolic medium (80%, v/v). The formation constants were determined for all binary complexes. The important infrared (IR) spectral bands corresponding to the active groups in the four ligands and the solid complexes under investigation were studied. The solid complexes have been synthesized and studied by thermogravimetric analysis. The thermal dehydration and decomposition of these complexes were studied kinetically using the integral method applying the Coats Redfern equation. It was found that the thermal decomposition of the complexes follow second order kinetics. The thermodynamic parameters of the decomposition are also reported. The electronic absorption spectra of the investigated ligands were carried out to determine the pK(a) values spectrophotometrically. PMID- 17084105 TI - Specific heat and frequency shifts for the translational modes in ammonia solid I close to phase transition. AB - The specific heat C(p) is correlated to the frequency shifts (1/nu)( partial differentialnu/ partial differentialT)(p) in ammonia solid I close to the melting point. Our calculated Raman frequencies for the translational modes of nu(T) (100 cm(-1)) and nu(T) (130 cm(-1)), are used for this correlation for the pressures of 0, 1.93 and 3.07 kbar in this crystalline system. We obtain that the specific heat varied linearly with the frequency shifts for those pressures studied and values of the slope dP(m)/dT were extracted. The observed behaviour of the ammonia solid I near the melting point is explained on the basis of our spectroscopic modification of the first Pippard relation. PMID- 17084106 TI - Toll road for Toxoplasma gondii: the mystery continues. AB - Toll-like receptors (TLRs) are considered to be essential for the initiation of immune responses against pathogens. Although myeloid differentiation factor 88 an adaptor molecule for most TLRs, is important for protection against Toxoplasma gondii, the TLR responsible for eliciting an immune response against this obligate intracellular pathogen remains unknown. A recent article reports that mice lacking TLR9 cannot develop severe inflammatory responses to T. gondii infection. The implications of this finding are discussed here. PMID- 17084108 TI - The 'A-dip' of diastolic mitral regurgitation: an unusual Doppler flow pattern in a patient with severe aortic insufficiency and complete heart block. AB - This is an unusual case of diastolic mitral regurgitation (MR) with a high diastolic velocity jet and prolonged jet duration related to a combination of acute severe aortic insufficiency and high-degree atrioventricular block. This case illustrates an interesting hemodynamic phenomenon with multiple transient decreases in the pressure gradient between the left ventricle and left atrium during diastole related to a temporary increase in left atrial pressure associated with atrial contraction. PMID- 17084109 TI - Natural language processing and visualization in the molecular imaging domain. AB - Molecular imaging is at the crossroads of genomic sciences and medical imaging. Information within the molecular imaging literature could be used to link to genomic and imaging information resources and to organize and index images in a way that is potentially useful to researchers. A number of natural language processing (NLP) systems are available to automatically extract information from genomic literature. One existing NLP system, known as BioMedLEE, automatically extracts biological information consisting of biomolecular substances and phenotypic data. This paper focuses on the adaptation, evaluation, and application of BioMedLEE to the molecular imaging domain. In order to adapt BioMedLEE for this domain, we extend an existing molecular imaging terminology and incorporate it into BioMedLEE. BioMedLEE's performance is assessed with a formal evaluation study. The system's performance, measured as recall and precision, is 0.74 (95% CI: [.70-.76]) and 0.70 (95% CI [.63-.76]), respectively. We adapt a JAVA viewer known as PGviewer for the simultaneous visualization of images with NLP extracted information. PMID- 17084107 TI - A female preponderance for chemically induced lupus in SJL/J mice. AB - Both spontaneous and chemically induced rodent models of autoimmune nephritis and autoantibody production have been explored to understand mechanisms involved in human systemic lupus erythematosus (SLE). While it has been known for decades that women are more susceptible than men to SLE, mechanisms underlying this female preponderance remain unclear. One chemically induced model involves injection of hydrocarbon oils such as pristane into otherwise normal mouse strains, which results in the development of autoantibodies and inflammation in organs such as kidney and liver. It is unknown whether lupus-like disease induced by chemicals would exhibit a sex bias in disease susceptibility. Here, we show that SJL/J female mice injected with pristane display greater mortality, kidney disease, serum anti-nuclear and anti-dsDNA antibodies than their male siblings. This is the first evidence that a female sex bias exists in a chemically induced lupus model. PMID- 17084110 TI - Acute hyperglycemia in uterine arteries from pregnant, but not non-pregnant mice, enhances endothelium-dependent relaxation. AB - Poorly controlled diabetes in pregnancy, characterized by hypo- and hyperglycemia, is associated with adverse outcomes. We hypothesized that aberrant glucose levels affect vascular function in pregnancy. The effects of glucose concentration on constriction and endothelium-dependent relaxation in uterine arteries of normal C57BL/6 mice were examined. Ex-vivo arteries from 18 pregnant and 14 non-pregnant mice were mounted on a wire myograph, constricted with phenylephrine and relaxed with incremental doses of acetylcholine, in physiological saline solution containing 5 mmol/L glucose. Arteries were then exposed to solutions with 2, 5, 8 or 12 mmol/L glucose for 30 min and constriction/relaxation repeated. On altering glucose concentrations to 2, 8 or 12 mmol/L, maximal constriction was increased in arteries from pregnant but not from non-pregnant mice (paired t-test, p<0.05). Endothelium-dependent relaxation was enhanced at 12 mmol/L glucose in arteries from pregnant (two-way ANOVA, p<0.01), but not from non-pregnant mice. Endothelium-dependent relaxation in the uterine artery was pre-dominantly mediated by a non-nitric oxide/non-prostanoid mechanism, with a smaller contribution from nitric oxide, and no prostanoid mediated relaxation. In summary, acute changes in glucose concentration alter both constriction and endothelium-dependent relaxation in uterine arteries of normal pregnant mice; these effects are unique to pregnancy. PMID- 17084111 TI - Electrochemical detection of short sequences related to the hepatitis B virus using MB on chitosan-modified CPE. AB - A novel electrochemical DNA biosensor based on methylene blue (MB) and chitosan modified carbon paste electrode (CCPE) for short DNA sequences and polymerase chain reaction (PCR) amplified real samples related to the hepatitis B virus (HBV) hybridization detection is presented. Differential pulse voltammetry (DPV) was used to investigate the surface coverage and hybridization event. The decrease in the peak current of MB, an electroactive label, was observed upon hybridization of probe with the target. Numerous factors affecting the target hybridization and indicator binding reaction are optimized to maximize the sensitivity. PMID- 17084113 TI - Identification and characterization of major flavonoids and caffeoylquinic acids in three Compositae plants by LC/DAD-APCI/MS. AB - In this study, a liquid chromatography/diode array detector-atmospheric pressure chemical ionization/mass spectrometry (LC/DAD-APCI/MS) was successfully developed to identify and characterize the main flavonoids and caffeoylquinic acids (CQAs) of three common Compositae plants (Chrysanthemum morifolium Raman, Artemisia annua, and Chrysanthemum coronarium) which have been used as herbal medicine. Identifications were performed by comparing the retention time, UV and mass spectra of samples with standards or/and earlier publications. The crude methanolic extracts of these plants were assayed directly using LC/MS without any further pretreatment. The proposed method is rapid and reproducible and is useful for characterization and evaluation of different plant flavonoids and CQAs. A total of 41 different flavonoids and 6 CQAs were identified and confirmed by APCI MS. The main components of three Compositae plants were also compared. Although there exist some similarities in the flavonoidic content of the leaf and flower of C. morifolium, significant variations in their varieties and concentrations were observed. Artemisia annua processes substantial amount of alkylated derivatives of flavones and Chrysanthemum coronarium contains only CQAs. These findings suggest that although all the plants studied are from the same Compositae family, their flavonoids and phenolic compositions are markedly different. The proposed method is useful for further chromatographic fingerprinting of plant flavonoids. PMID- 17084112 TI - A molecular characterization of spontaneous frameshift mutagenesis within the trpA gene of Escherichia coli. AB - Spontaneous frameshift mutations are an important source of genetic variation in all species and cause a large number of genetic disorders in humans. To enhance our understanding of the molecular mechanisms of frameshift mutagenesis, 583 spontaneous Trp+ revertants of two trpA frameshift alleles in Escherichia coli were isolated and DNA sequenced. In order to measure the contribution of methyl directed mismatch repair to frameshift production, mutational spectra were constructed for both mismatch repair-proficient and repair-defective strains. The molecular origins of practically all of the frameshifts analyzed could be explained by one of six simple models based upon misalignment of the template or nascent DNA strands with or without misincorporation of primer nucleotides during DNA replication. Most frameshifts occurred within mononucleotide runs as has been shown often in previous studies but the location of the 76 frameshift sites was usually outside of runs. Mismatch repair generally was most effective in preventing the occurrence of frameshifts within runs but there was much variation from site to site. Most frameshift sites outside of runs appear to be refractory to mismatch repair although the small number of occurrences at most of these sites make firm conclusions impossible. There was a dense pattern of reversion sites within the trpA DNA region where reversion events could occur, suggesting that, in general, most DNA sequences are capable of undergoing spontaneous mutational events during replication that can lead to small deletions and insertions. Many of these errors are likely to occur at low frequencies and be tolerated as events too costly to prevent or repair. These studies also revealed an unpredicted flexibility in the primary amino acid sequence of the trpA product, the alpha subunit of tryptophan synthase. PMID- 17084114 TI - Comparison of the relative benefits of 2 versus 10 days of soft collar cervical immobilization after acute whiplash injury. AB - OBJECTIVE: To investigate the effects of 2-day and 10-day immobilization of the cervical spine on pain, range of motion (ROM), and disability of patients with Quebec Task Force (QTF) grade II whiplash injuries. DESIGN: Randomized controlled trial. SETTING: University hospital emergency department. PARTICIPANTS: Seventy patients with acute QTF grade II whiplash injuries. INTERVENTIONS: At the intake examination within 24 hours after the whiplash trauma, the patients were randomized to 2 therapy groups (2-d or 10-d immobilization with a soft cervical collar). All patients received pain drugs (nonsteroidal anti-inflammatory drugs) and after 7 days, all patients started a standardized physiotherapy program 2 to 3 times a week. MAIN OUTCOME MEASURES: Patients' pain and disability scores were assessed using visual analog scales and ROM was assessed using a goniometer. All parameters were measured within 24 hours after injury and after 2 and 6 months. RESULTS: After 2 months, the different periods of immobilization (2d or 10d) were associated with comparable improvements in pain symptoms (median, 4.60 vs 4.65), ROM (median, 100.0 degrees vs 117.5 degrees ), and disability score (median, 4.90 vs 5.15). No statistically significant differences could be identified between the 2 treatment groups. After 6 months, persistent pain was reported by 4 patients in each group (12.5%). CONCLUSIONS: In patients with QTF grade II whiplash injuries, there is no short- or long-term difference between 2-day and 10-day immobilization with a cervical collar in terms of pain, ROM, or disability. PMID- 17084115 TI - Outcomes of a weight-bearing rehabilitation program for patients diagnosed with patellofemoral pain syndrome. AB - OBJECTIVE: To determine the effects of a weight-bearing rehabilitation program on quadriceps and gluteus medius electromyographic activity, pain, and function in subjects diagnosed with patellofemoral pain syndrome (PFPS). DESIGN: Pretest and posttest 6-week intervention study. SETTING: Musculoskeletal research laboratory. PARTICIPANTS: Fourteen subjects diagnosed with PFPS and 14 healthy control subjects volunteered to participate in this study. No subjects withdrew from the study because of adverse effects. INTERVENTION: Subjects diagnosed with PFPS participated in a 6-week rehabilitation program. The rehabilitation program consisted of weight-bearing exercises that focused on strengthening the quadriceps and hip abductor musculature. MAIN OUTCOME MEASURES: Electromyographic onsets of the vastus medialis oblique (VMO) and vastus lateralis and onset and duration of the gluteus medius were collected during a stair-stepping task that was performed during the pretest and posttest. A visual analog scale (VAS) and Functional Index Questionnaire (FIQ) were administered at pretest and posttest and each week of the intervention. RESULTS: Vastus lateralis and VMO onset timing differences (vastus lateralis electromyographic onset minus VMO electromyographic onset) and VAS and FIQ scores significantly improved for patients diagnosed with PFPS. Vastus lateralis and VMO onset timing in the PFPS group were significantly different from those in the control group at baseline and were not significantly different from the control group after the intervention. We did not find differences in gluteus medius onsets or duration of activity. CONCLUSIONS: Subjects diagnosed with PFPS responded favorably and quickly to a therapeutic exercise program that incorporated quadriceps and hip musculature strengthening. The efficacy of the therapeutic exercise program used in this study should be further investigated in a larger subject population. PMID- 17084116 TI - Effects of disease severity on response to lateral wedged shoe insole for medial compartment knee osteoarthritis. AB - OBJECTIVE: To determine the effects of lateral wedged insoles on knee kinetics and kinematics during walking, according to radiographic severity of medial compartment knee osteoarthritis (OA). DESIGN: A prospective case control study of patients with medial compartment OA of the knee. SETTING: Gait analysis laboratory in a university hospital. PARTICIPANTS: Forty-six medial compartment knees with OA of 23 patients with bilateral disease and 38 knees of 19 age matched healthy subjects as controls. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: We measured the peak external adduction moment at the knee during the stance phase of gait and the first acceleration peak after heel strike at the lateral side of the femoral condyles. Kellgren and Lawrence grading system was used for radiographic assessment of OA severity. RESULTS: The mean value of peak external adduction moment of the knee was higher in OA knees than the control. Application of lateral wedged insoles significantly reduced the peak external adduction moment in Kellgren-Lawrence grades I and II knee OA patients. The first acceleration peak value after heel strike in these patients was relatively high compared with the control. Application of lateral wedged insoles significantly reduced the first acceleration peak in Kellgren-Lawrence grades I and II knee OA patients. CONCLUSIONS: The kinetic and kinematic effects of wearing of lateral wedged insoles were significant in Kellgren-Lawrence grades I and II knee OA. The results support the recommendation of use of lateral wedged insoles for patients with early and mild knee OA. PMID- 17084117 TI - Factors influencing involvement in research and career choice: a survey of graduating physical medicine and rehabilitation residents. AB - OBJECTIVE: To assess the extent to which physical medicine and rehabilitation (PM&R) residents' involvement in research and selection of postgraduate practice may be related to residency program research requirements and support for presentations. DESIGN: Internet-based survey. SETTING: Anonymous access to Web form. PARTICIPANTS: PM&R residents graduating in June 2004 and 2005. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Involvement in research, postgraduate practice setting and planned research effort, residency research requirement, and support for scientific presentations. RESULTS: One hundred sixty (24%) of the 657 graduating residents, representing 25 states, the District of Columbia, and Puerto Rico, responded. Eighty-five percent indicated research involvement, with 74% reporting a research requirement and 85% residency program financial support for presentations. On average, respondents planned to devote 7% (95% confidence interval, 5%-9%) of their time to research once in practice. There was a statistically significant association between the existence of a research requirement and involvement in research (P < .001). However, there was no evidence of a statistically significant association between either the existence of a research requirement or presentation support and the selection of an academic career. There were no significant regional disparities for the existence of a research requirement, travel support, or number of presentations. However, there was a statistically significant regional difference in the proportion of postgraduate practice time that the respondents planned to devote to research. CONCLUSIONS: A requirement for PM&R residents to be involved in research may influence research activity during residency but may not be associated with selection of an academic or research-oriented practice. PMID- 17084118 TI - The potential use of axial spinal unloading in the treatment of adolescent idiopathic scoliosis: a case series. AB - OBJECTIVE: To assess potential benefits of axial spinal unloading (LTX 3000 Lumbar Rehabilitation System) over a brief 3-month period. DESIGN: Before-after pilot study. SETTING: University research laboratory. PARTICIPANTS: Five adolescent girls with scoliosis. INTERVENTIONS: Three laboratory sessions: (1) initial baseline, (2) immediately after 3-month treatment period (axial unloading by using LTX 3000 for two 10-minute treatments daily), and (3) 1 month posttreatment. MAIN OUTCOME MEASURES: Initial baseline postural data were obtained from 2 sets of radiographs (standing anteroposterior [AP] and lateral, sitting AP and lateral), back range of motion measurements, and numeric pain scales. The following were assessed: static postural changes; potential functional benefits; and therapeutic compliance. RESULTS: All subjects elicited reductions in lumbar Cobb angles immediately after 3 months of treatment; initial average scoliotic curves of 13.7 degrees were reduced 42% to 8 degrees (alpha = .05, P = .004). Additionally, such reductions were evident 1 month posttreatment; average original curves were reduced by 27%. Subjects' range of motion and lumbar lengthening were not significantly altered by this therapeutic protocol. Reported subject compliance was high (95%). CONCLUSIONS: The LTX 3000 is a potential adjunct therapy for the treatment of adolescent scoliosis. This therapy resulted in curvature reductions and was widely accepted by subjects that were compliant with self-administration. PMID- 17084119 TI - Effects of lumbar stabilization using a pressure biofeedback unit on muscle activity and lateral pelvic tilt during hip abduction in sidelying. AB - OBJECTIVE: To assess the effects of lumbar spine stabilization using a pressure biofeedback unit on the electromyographic activity and angle of lateral pelvic tilt during hip abduction in a sidelying position. DESIGN: Comparative, repeated measures study. SETTING: University research laboratory. PARTICIPANTS: Eighteen able-bodied volunteers (9 men, 9 women) with no history of pathology. INTERVENTION: Subjects were instructed to perform hip abduction in a sidelying position in both the preferred hip abduction (PHA) and hip abduction with lumbar stabilization (HALS). A pressure biofeedback unit was used for lumbar stabilization. MAIN OUTCOME MEASURES: Surface electromyography was recorded from the quadratus lumborum, gluteus medius, internal oblique, external oblique, rectus abdominis, and multifidus muscles. Kinematic data for lateral pelvic tilt angle were measured using a motion analysis system. Dependent variables were examined with 2 (PHA vs HALS) x 2 (men vs women) analysis of variance. RESULTS: Significantly decreased electromyographic activity in the quadratus lumborum (PHA, 60.39% +/- 15.62% of maximum voluntary isometric contraction [MVIC]; HALS, 27.90% +/- 13.03% of MVIC) and significantly increased electromyographic activity in the gluteus medius (PHA, 25.03% +/- 10.25% of MVIC; HALS, 46.06% +/- 21.20% of MVIC) and internal oblique (PHA, 24.25% +/- 18.10% of MVIC; HALS, 44.22% +/- 20.89% of MVIC) were found when the lumbar spine was stabilized. Lateral pelvic tilt angle (PHA, 13.86 degrees +/- 4.66 degrees; HALS, 5.55 degrees +/- 4.16 degrees) was decreased significantly when the lumbar spine was stabilized. In women the electromyographic activity (percentage of MVIC) in gluteus medius, external oblique, and rectus abdominis was significantly higher than that observed in men. CONCLUSIONS: With lumbar stabilization, the gluteus medius and internal oblique activity was increased significantly, and the quadratus lumborum activity was decreased significantly, causing reduced lateral pelvic tilt in a sidelying position. These results suggest that hip abduction with lumbar stabilization is useful in excluding substitution by the quadratus lumborum. PMID- 17084120 TI - Bone mineral density in hip-fracture patients with Parkinson's disease: a case control study. AB - OBJECTIVE: To investigate bone mineral density (BMD) levels in patients with Parkinson's disease (PD) who sustained a hip fracture. DESIGN: Case-control study. SETTING: Rehabilitation hospital in Italy. PARTICIPANTS: We investigated 831 out of 887 white patients consecutively admitted to a rehabilitation hospital because of an original hip fracture resulting from a fall. Twenty-eight (3.37%) of the 831 patients were affected by PD. Twenty-eight controls matched for sex, age, and hip-fracture type (cervical or trochanteric) were found among the 803 non-PD patients. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: BMD was assessed by dual-energy x-ray absorptiometry (DXA) at the unfractured femur. Five sites were investigated in each subject: total proximal femur, femoral neck, trochanter, intertrochanteric area, and Ward's triangle. DXA scan was performed a mean +/- standard deviation of 22.2 +/- 7.8 days after fracture occurrence in the 28 patients and 22.0 +/- 5.3 days after fracture occurrence in the 28 controls. RESULTS: BMD expressed as a T score did not differ significantly between the 28 PD patients and the 28 controls, whereas z score in the PD patients was significantly lower than 0 +/- 1 in the age- and sex-matched general population at 4 of the 5 sites of BMD assessment. CONCLUSIONS: A sample of PD fallers who sustained a hip fracture had femoral BMD levels similar to those found in matched hip-fracture fallers who did not suffer from PD and significantly lower than those found in the matched reference population. PMID- 17084121 TI - Kinematic analysis of upper limbs and trunk movement during bilateral movement after stroke. AB - OBJECTIVE: To compare the kinematics of the upper limbs and trunk during unilateral and parallel bilateral tasks in subjects with hemiparesis and control subjects. DESIGN: Comparative study. SETTING: Geriatric center offering rehabilitation services. PARTICIPANTS: Convenience sample of 15 persons (age, 69.4 +/- 12.0 y; > or = 3 mo poststroke) recruited in a geriatric center with rehabilitation services, and 13 control persons (67.8 +/- 7.5 y) participated in the study. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Unilateral and bilateral movements toward 1 or 2 targets located beyond arm's length and positioned in 3 directions. Angular changes of both upper limbs and trunk were characterized in the sagittal, frontal, and horizontal planes. RESULTS: During the bilateral task, the deficits of the kinematic joints of the paretic upper limb persisted in subjects with hemiparesis as compared with the corresponding upper limb in the control subjects (abduction shoulder: subjects with hemiparesis, 5.7 degrees +/- 5.3 degrees; control subjects, 0.7 degrees +/- 4.8 degrees; extension elbow: subjects with hemiparesis, 38.2 degrees +/- 14.2 degrees; control subjects, 52.8 degrees +/- 12.5 degrees) with a marked flexion of the trunk (subjects with hemiparesis, 33.7 degrees +/- 8.7 degrees; control subjects, 26.8 degrees +/- 5.8 degrees). The elbow extension of the nonparetic upper limb was reduced (subjects with hemiparesis, 41.0 degrees +/- 13.6 degrees; control subjects, 52.8 degrees +/- 12.5 degrees). CONCLUSIONS: The use of parallel bilateral reaching tasks and placing movements of the upper extremities in the subjects with hemiparesis contributed an increase in the trunk flexion rather than improve the motor performance of the paretic upper limb, especially with regard to increasing elbow extension. PMID- 17084122 TI - Development of the Hand Active Sensation Test: reliability and validity. AB - OBJECTIVE: To develop and establish the reliability and validity of a new quantitative functional measure of haptic perception in the hand, the Hand Active Sensation Test (HASTe). DESIGN: Reliability was assessed by test-retest sessions. Validity was assessed via discriminant analysis, concurrent validity with 2-point discrimination and wrist position test, and receiver operating characteristic (ROC) curve construction. SETTING: Subject preference. PARTICIPANTS: Heterogeneous sample of 28 stroke survivors and 28 individually matched controls. INTERVENTION: Subjects used 1 hand to manipulate HASTe objects that vary by weight or texture to complete 18 match-to-sample trials. MAIN OUTCOME MEASURES: Two-point discrimination threshold, Wrist Position Sense Test (WPST) average error, and HASTe accuracy score. RESULTS: Test-retest reliability was strong (intraclass correlation coefficient model 3,1 = .77). The HASTe score significantly discriminated the groups (t = 8.3, P < .001) and correlated with 2 point discrimination (r = -.67, P < .001) and WPST (r = -.60, P < .001). ROC curve area was .94 for test 1 and .92 for the average of 2 tests. CONCLUSIONS: The HASTe is a reliable and valid functional measure of haptic perception, appears to detect impairment of haptic perception even in stroke survivors with no reported sensory deficits, and may provide valuable quantitative clinical data about complex sensory loss and hand function after stroke. PMID- 17084123 TI - Development of a multidimensional balance scale for use with functionally independent older adults. AB - OBJECTIVE: To develop and evaluate the validity and reliability of a multidimensional balance scale-the Fullerton Advanced Balance (FAB) scale suitable for use with functionally independent older adults. DESIGN: Psychometric evaluation of the scale's content and convergent validity, test-retest and intra- and interrater reliability, and internal rater consistency. SETTING: Urban community. PARTICIPANTS: Forty-six community-residing older adults (mean +/- standard deviation, 75 +/- 6.2 y), with (n = 31) and without identified balance problems (n = 15), participated in the study. Four physical therapists with expertise in the assessment and treatment of balance disorders in older adults also participated in the content validity and/or reliability phases of the study. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Spearman rank correlation coefficients for convergent validity, test-retest, intra- and interrater reliability, and homogeneity coefficient values for rater consistency. RESULTS: Test-retest reliability for the total balance scale score was high (rho = .96). Interrater reliability for total score ranged from .94 to .97 whereas intrarater reliability coefficients ranged from .97 to 1.00. Homogeneity (H) coefficients were greater than .90 for 6 of the 10 individual test items and all 10 test items had H coefficients of greater than .75 for both rating sessions. CONCLUSIONS: Preliminary results suggest that the FAB scale is a valid and reliable assessment tool that is suitable for use with functionally independent older adults residing in the community. PMID- 17084124 TI - Control of balance differs after knee or ankle fatigue in older women. AB - OBJECTIVE: To examine the effects of acute isokinetic knee or ankle fatigue on control of static and dynamic balance in older women. DESIGN: Pretest and posttest. SETTING: University research laboratory. PARTICIPANTS: Eighteen healthy, community-dwelling older women (age, 77 +/- 6 y) with no history of falling. INTERVENTIONS: Measurements of static and dynamic balance control before and after isokinetically fatiguing the ankle plantar- and dorsiflexors or knee extensors and flexors in separate sessions. MAIN OUTCOME MEASURES: Performance on 3 clinical assessments of control of balance: modified Functional Reach Test (mFRT), Lower-Extremity Reach Test (LERT), and Single-Limb Stance Time Test (SLSTT). RESULTS: Balance declined in the mFRT after fatigue to each joint, with no significant difference in the magnitude of change between joints. Control of balance during the LERT decreased significantly only after knee fatigue, and control of balance during the SLSTT was significantly reduced only after ankle fatigue. CONCLUSIONS: Balance performance after acute isokinetic muscular fatigue to the knee or ankle is specific to the muscle groups fatigued and the balance tests used. PMID- 17084125 TI - Development of a position-specific index of muscle strength to be used in stroke evaluation. AB - OBJECTIVE: To develop a position-specific index of muscle strength for individuals with stroke. DESIGN: Cross-sectional design. SETTING: A major teaching hospital in a Canadian urban city. PARTICIPANTS: Sixty-three patients with poststroke onset between 3 and 12 months. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: The muscle strength of the lower-extremity muscles was tested bilaterally in multiple positions using hand-held dynamometry. RESULTS: A principal components analysis resulted in grouping the muscles of the affected and unaffected sides of the gravity related and gravity eliminated positions into 5 indices. The 5 indices were moderately to highly correlated (r2 range, .59-.81) with each other and so were combined into 1 global index. The gravity related muscle strength on the affected side was, on average, 85% of the unaffected side (range, 37%-157%); the gravity eliminated muscle strength of the affected side was, on average, 92% of the unaffected side (range, 53%-121%). CONCLUSIONS: This study resolves the methodologic issue of how to summarize multiple data points that relate to one construct, namely, strength of different muscle groups assessed in several positions. PMID- 17084126 TI - Predicting recovery of upper-body dressing ability after stroke. AB - OBJECTIVE: To identify predictors of the recovery of independent dressing ability after stroke. DESIGN: Prospective cohort study. SETTING: Rehabilitation unit at a university hospital. PARTICIPANTS: Sixty-three consecutive stroke patients were enrolled in the study. Twelve patients were not able to complete the study because they were discharged or transferred to another hospital before study completion. INTERVENTION: Fifty-one patients underwent and completed 15 days of dressing training based on the time-delay method, which included the 10 component actions of upper-body dressing and 4 cues given by therapists. MAIN OUTCOME MEASURES: The dressing item of the FIM instrument, Brunnstrom motor recovery stages, presence or absence of deep and tactile sensation, Rey-Osterrieth complex figure test, Kohs block design test, body image test, Weintraub cancellation task, and presence or absence of the visual extinction phenomenon and the motor impersistence phenomenon. RESULTS: The FIM upper-body dressing item score and the cancellation task score at the start of training were significantly better in patients who achieved independence in dressing within 15 training days than in patients who did not (P < .05). The motor impersistence phenomenon was found less frequently among patients who achieved independence in upper-body dressing than among patients who did not (P < .05). However, logistic regression analysis showed that only the FIM score for upper-body dressing on the first day of training was a significant independent predictor of dressing ability at the end of training (odds ratio, 4.33; 95% confidence interval, 1.51-12.37). The receiver operating characteristic curve indicated that a cutoff score of 3 would provide the best balance between sensitivity and specificity for the FIM upper-body dressing item. The positive predictive value of this cutoff score was .90, and the negative predictive value was .70. CONCLUSIONS: Our findings indicate that the FIM upper-body dressing score on the first day of dressing training is an independent predictor of recovery of upper-body dressing ability after stroke. PMID- 17084127 TI - Weight gain in children with hypertonia of cerebral origin receiving intrathecal baclofen therapy. AB - OBJECTIVE: To identify the pattern of weight change in children receiving intrathecal baclofen (ITB) therapy. DESIGN: A retrospective medical chart review was conducted to identify weight status before and after ITB pump implantation. SETTING: Tertiary care children's hospital and academic medical center. PARTICIPANTS: All children and adolescents with hypertonia of cerebral origin who were younger than 19 years of age at the time of pump placement and followed in our pediatric baclofen pump program. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: A linear mixed model was used to examine the rate of change in weight (weight-gain velocity) before and after surgery. Weight change was adjusted for age, sex, functional level determined by the Gross Motor Functional Classification System, tube feeding, dystonia, and other comorbidities. RESULTS: The average weight-gain velocity was 2.32 kg/y presurgery and 2.93 kg/y postsurgery, adjusted for potential confounders. The 0.61 kg/y increase in weight gain velocity attained statistical significance (P = .028). CONCLUSIONS: Although excessive weight gain is not a common problem in children with cerebral origin spasticity, increased weight-gain velocity is prevalent in children receiving ITB therapy. Health care providers may anticipate a welcome weight gain in the underweight child. This consequence should be considered when managing children receiving ITB therapy, and health care providers must appropriately intervene to prevent excessive weight gain. Further studies exploring the reasons for the increased weight-gain velocity are warranted. PMID- 17084128 TI - Intrathecal baclofen management of poststroke spastic hypertonia: implications for function and quality of life. AB - OBJECTIVES: To evaluate the impact of intrathecal baclofen (ITB) on function and quality of life (QOL) and to obtain efficacy and safety data in poststroke spastic hypertonia. DESIGN: Prospective open-label multicenter trial with follow up at 3 and 12 months. SETTING: Twenty-four stroke treatment centers in the United States. PARTICIPANTS: Ninety-four stroke participants (age range, 24-82 y) with spastic hypertonia. Seventy-four participants underwent ITB pump implantation. INTERVENTION: Participants were implanted with an ITB pump. MAIN OUTCOME MEASURES: FIM instrument and QOL (Sickness Impact Profile [SIP]) changes, spastic hypertonia (Ashworth Scale), and safety. RESULTS: FIM scores improved overall in repeated-measures analysis of variance (ANOVA) (P = .005) and by 3.00 +/- 7.69 (P = .001) at 3 months and by 2.86 +/- 10.13 (P = .017) at 12 months. Significant improvements in SIP scores were noted overall (repeated-measures ANOVA, P < .001) and at 3 (P = .003) and 12 months (P < .001). The combined average Ashworth Scale score of the upper and lower limbs decreased by 1.27 +/- 0.76 (P < .001) at 3 months and by 1.39 +/- 0.73 (P < .001) at 12 months from baseline, which was significant overall (repeated-measures ANOVA, P<.001). Strength in the unaffected side did not change overall (repeated-measures ANOVA, P = .321) or at either 3 (P = .553) or 12 months (P = .462). Minimal adverse events and device complications were reported. CONCLUSIONS: There was significant improvement in function, QOL, and spastic hypertonia at 3 and 12 months after implant, without adversely affecting muscle strength of the unaffected limbs. Data suggest that ITB therapy is a safe and efficacious treatment for spastic hypertonia resulting from stroke. PMID- 17084129 TI - Data-based models of how pressure ulcers develop in daily-living contexts of adults with spinal cord injury. AB - OBJECTIVE: To examine the daily-lifestyle influences on the development of pressure ulcers in adults with spinal cord injury (SCI). DESIGN: Qualitative investigation using in-depth interviewing and participant observation. SETTING: Participants were studied in their homes and other naturalistic contexts. PARTICIPANTS: Twenty men and women of diverse ethnicities with paraplegia or tetraplegia who were recruited at a pressure ulcer management clinic in a large rehabilitation facility. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Detailed descriptive information pertaining to the development of recurring pressure ulcers in relation to participants' daily routine and activity, personal choices, motivating influences, lifestyle challenges, and prevention techniques and strategies. RESULTS: The daily-lifestyle influences on pressure ulcer development in adults with SCI can be described through various models that vary in complexity, depending on whether they incorporate individualization, interrelations among modeled elements, situational specificity, and/or temporal comprehensiveness. Ulcers are most likely to develop when a person with a relatively high-risk background profile is exposed to an equilibrium-disrupting change event that culminates in a specific pressure ulcer risk episode. CONCLUSIONS: The results underscore the significant degree of complexity and individualization that characterize the emergence of pressure ulcers in daily life contexts. Prevention efforts should therefore incorporate attention to the unique constellation of circumstances that comprise a person's everyday life. PMID- 17084130 TI - The use of the Gaussian curve fitting method for scintigraphic measurements of the swallowing process in healthy subjects: implications for evaluation of dysphagia. AB - OBJECTIVE: To present the results of scintigraphic evaluation, using the Gaussian curve fitting method, via 3 parameters of oropharyngeal swallow: (1) pharyngeal transit time, (2) premature pharyngeal entry, and (3) postswallow pharyngeal stasis while ingesting liquid. DESIGN: A descriptive study. SETTING: A rehabilitation hospital affiliated with a medical university. PARTICIPANTS: Eighteen healthy subjects. INTERVENTION: All 18 subjects received scintigraphic swallow examination to evaluate dynamic swallow process of 5 mL of liquid. MAIN OUTCOME MEASURES: The Gaussian curve fitting method was used to calculate the pharyngeal transit time, premature pharyngeal entry, and postswallow pharyngeal stasis. RESULTS: The mean pharyngeal transit time was .71 seconds. The maximal percentage of premature pharyngeal entry was 3%. The maximal percentage of postswallow pharyngeal stasis was 9%. CONCLUSIONS: The Gaussian curve fitting can be used as an objective and time-saving method to calculate the parameters in scintigraphic swallowing examination. Our results approximate other researchers' reports. PMID- 17084131 TI - Comparison of gastrostomy tube replacement verification using air insufflation versus gastrograffin. AB - OBJECTIVE: To compare the safety of air insufflation as a contrast medium for percutaneous endoscopic gastrostomy (PEG) tube replacement verification with that of the more traditional method using positive contrast material (gastrograffin) under fluoroscopic guidance. DESIGN: Retrospective case series comparing 2 radiographic procedures used for PEG tube verification at a rehabilitation hospital. The first series reviewed studies of patients over 1 year, during which the method of choice for PEG tube replacement verification was with air insufflation. The second series reviewed studies of patients over the subsequent year, during which the method of choice for PEG tube replacement verification was with gastrograffin. SETTING: An inpatient rehabilitation hospital. PARTICIPANTS: Nineteen patients and 29 PEG tube replacements were identified that used air insufflation and 17 patients and 19 PEG tube replacements were identified that used gastrograffin. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Medical complications in a 2-week follow-up period as a result of the verification procedures. RESULTS: There were no cases in either of the replacement procedures in which the PEG tube was found to not be in the gastrointestinal system. There was also no evidence at 2-week follow-up that any of the patients' complications were the result of either procedure. CONCLUSIONS: The air contrast procedure appears to be a relatively safe alternative procedure for PEG tube replacement verification. PMID- 17084132 TI - Gait performance in an original biologic reconstruction of proximal femur in a skeletally immature child: a case report. AB - Biologic reconstruction of the femoral head by a sophisticated autotransplantation of the proximal growing fibula associated with a massive bone allograft has been performed in a 4-year-old girl affected by Ewing's sarcoma. The child was treated in 1997 and then followed: clinical and functional tests were performed 2, 3, 4, 6, and 7 years after surgery. Gait and specific motor tasks were assessed by means of motion analysis instruments. The patient was followed by a specific rehabilitation program, aimed at controlling load on the treated limb, resuming good muscle function, and recovering a physiological pattern of movement during daily routine activities. The outstanding radiographic evolution 8 years after surgery with the peroneal head progressively remodeled as a femoral head, and the more than satisfactory gait pattern observed at last follow-up, makes this study a keystone of experience and knowledge to be applied to other patients. PMID- 17084133 TI - A novel instrument for quantifying grip activity during handwriting. AB - OBJECTIVE: To describe and exemplify a noninvasive instrumentation system that uniquely records grip forces exerted on the barrel of the writing utensil along with typical temporal and kinematic parameters. DESIGN: Exploratory case series. SETTING: A tertiary care, pediatric rehabilitation hospital. PARTICIPANTS: Convenience sample of 6 primary school children with spastic hemiplegic cerebral palsy (CP) with documented fine motor difficulties and 6 children with no known handwriting difficulties. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Grip and normal forces during handwriting. RESULTS: The instrumentation revealed nontrivial correlations between normal and grip forces (0.55 +/- 0.16), a temporal delay between normal and grip forces (97.7 +/- 16 ms), and a consistent grip-to-normal force ratio (4.3 +/- 1.5), across all participants. Grip force distributions agreed intuitively with qualitative observations of individual grasps of the writing utensil. Further, 5 new parameters derived from grip force measures statistically differentiated between able-bodied children and those with hemiplegic CP. CONCLUSIONS: The proposed instrumentation provides new insight into grip and normal force coordination and has the potential to uncover discriminatory grip force characteristics between writers with and without handwriting difficulties. Further study with larger populations is required to define functional ranges for the new grip measures. PMID- 17084134 TI - Understanding multiple sclerosis fatigue: a synthesis of biological and psychological factors. PMID- 17084135 TI - Fatigue and social impairment in multiple sclerosis: the role of patients' cognitive and behavioral responses to their symptoms. AB - OBJECTIVES: The aim of this study was to test whether the way in which multiple sclerosis (MS) patients interpret and respond to their symptoms may help to explain their experience of fatigue and social impairment. METHOD: In a cross sectional study, 149 patients with a definite diagnosis of MS completed validated questionnaires that measured their symptom interpretation and behavioral response, fatigue, mood, and social adjustment. A neurologist assessed their level of neurological impairment. RESULTS: Regression analyses revealed that patients' cognitive interpretations of their symptoms, such as a tendency to attribute a wide range of symptoms to their MS and feelings of embarrassment about symptoms, were significantly associated with fatigue and social adjustment over and above neurological impairment, remission status, and mood. The behavioral variables including an all-or-nothing response to symptoms and rest/avoidance behaviours were also strongly associated with fatigue and social impairment. The degree of neurological deficit was not associated with either physical or mental fatigue, although it was associated with social impairment. Depression, anxiety, and remission status were also related to fatigue, and depression to social impairment. CONCLUSION: These findings suggest that treatment approaches that challenge unhelpful cognitive interpretations and behavioural responses to symptoms, and address anxiety and depression, may be important in improving the overall management of fatigue in MS. PMID- 17084136 TI - Well-being in patients with chronic fatigue syndrome: the role of acceptance. AB - OBJECTIVE: Research in chronic pain patients has shown that accepting the chronic nature of their illness is positively related to quality of life. The aim of this study was to investigate whether acceptance is also associated with better well being in patients suffering from chronic fatigue syndrome (CFS). METHODS: Ninety seven patients completed a battery of questionnaires measuring fatigue, functional impairment, psychological distress, and acceptance. RESULTS: Results indicated that acceptance has a positive effect upon fatigue and psychological aspects of well-being. More specifically, acceptance was related to more emotional stability and less psychological distress, beyond the effects of demographic variables, and fatigue severity. CONCLUSION: We suggest that promoting acceptance in patients with CFS may often be more beneficial than trying to control largely uncontrollable symptoms. PMID- 17084137 TI - Long-term efficacy of cognitive-behavioral therapy by general practitioners for fatigue: a 4-year follow-up study. AB - OBJECTIVE: In an earlier study, we found that cognitive-behavioral therapy (CBT) delivered by general practitioners (GPs) for fatigue among employees on sick leave was not effective after 12 months. In this study we aim to assess the long term efficacy of CBT by GPs for fatigue. It was hypothesized that the intervention could prevent deterioration as well as relapse of fatigue complaints and relapse into absenteeism in the long term. METHODS: Patients who participated in the original randomized controlled trial were followed up 4 years later. Fatigue and absenteeism were the main outcomes. RESULTS: Fatigue and absenteeism were high in the intervention and control groups at the 4-year follow-up. There was no significant difference between the intervention group and the control group on fatigue and absenteeism. The intervention group however tended toward less-favorable outcomes as compared with the control group. CONCLUSIONS: Like that of chronic fatigue syndrome, the prognosis of less-advanced fatigue is rather poor. CBT delivered by GPs is not effective in the long term. PMID- 17084138 TI - Psychiatric manifestations and personality traits associated with compliance with glaucoma treatment. AB - OBJECTIVE: The aim of this study was to assess the impact of psychological distress and personality traits on self-rated compliance with glaucoma treatment. METHODS: One hundred patients with primary open-angle glaucoma participated in the study. General Health Questionnaire, Symptom Distress Checklist, Center for Epidemiological Studies Depression Scale, Defense Style Questionnaire, and Hostility and Direction of Hostility Questionnaire were used. RESULTS: Forty-two percent of patients with glaucoma classified as "noncompliers," those who omitted more than two doses per week. Noncompliers presented more severe symptoms of glaucoma. Depression was found to be associated with poor compliance, whereas adoption of immature defensive style further increased the risk for noncompliance with glaucoma treatment. CONCLUSIONS: Depression is associated with self-reported noncompliance with glaucoma treatment, whereas certain personality traits are involved in the increased risk for noncompliance. Further assessment of the depressive feelings by an ophthalmologist and treatment of depression as well as proper psychotherapeutic approaches for maladaptive personality features could be an essential strategy to diminish compliance problems. PMID- 17084139 TI - Socioeconomic differences in incident depression in older adults: the role of psychosocial factors, physical health status, and behavioral factors. AB - OBJECTIVE: The objective of this study was to examine the association between socioeconomic status (SES) and the onset of depression in older adults and to determine the relative contribution of psychosocial factors, physical health status, and behavioral factors in explaining this link. METHODS: Data were from 2593 men and women, aged 55-85 years, participating in the Longitudinal Aging Study Amsterdam. Two indicators of SES were used: education and income. The onset of depression was measured over 9 years of follow-up. RESULTS: Adjusted hazard ratios of incident depression were significantly higher in those with low education and low income. Psychosocial factors explained on average 16% of the SES differences in incident depression, physical health status on average 7%, and behavioral factors less than 5%. CONCLUSION: In older adults, low SES predicted the incidence of depression. Part of this association was explained by psychosocial factors and physical health status. PMID- 17084140 TI - Age is strongly associated with alexithymia in the general population. AB - OBJECTIVE: We studied the prevalence of alexithymia, its distribution in different age groups in a wide age range, its association with sociodemographic and health-related variables, and its co-occurrence with depression. METHODS: The study forms part of the Health 2000 Study. The original sample comprised 8028 subjects representing the general adult population of Finland. Alexithymia was measured with the 20-item Toronto Alexithymia Scale (TAS-20), and depression was measured with the 21-item Beck Depression Inventory. Altogether, 5454 participants filled in TAS-20 in their mother tongue. RESULTS: The prevalence of alexithymia was 9.9%. Men (11.9%) were more commonly alexithymic than women (8.1%). Alexithymia was associated with male gender, increasing age, low educational level, poor perceived health, and depression. CONCLUSIONS: The findings were in line with earlier population studies. For the first time, it was possible to analyze the prevalence of alexithymia in a wide age range (30-97 years). International comparative studies are needed. PMID- 17084141 TI - Posture, movement patterns, and body awareness in women with chronic pelvic pain. AB - OBJECTIVE: Chronic pelvic pain (CPP) is a common cause of infirmity but is still poorly understood. We studied the clinical characteristics, including body awareness, of 60 women with this diagnosis compared to those of healthy controls in an effort to understand its pathophysiology and to develop a more efficient treatment protocol. METHODS: After prior gynecologic and psychometric evaluation, the women were examined with the Standardized Mensendieck Test to evaluate posture and movement patterns. Pain history and pain score were obtained, and patterns of muscular density, elasticity, and tenderness were determined by palpation. The body awareness of patients was assessed through clinical evaluation. RESULTS: Seventy percent of the patients had a history of trauma or infection of the genitourinary region. The average pain score (+/-S.D.) on a scale from 0 to 10 was 6.01+/-1.60. Nearly all patients had a dissociative pattern, with a lack of contact and control of large body regions. All scores for posture and movement patterns were significantly worse in patients than in healthy women. CONCLUSION: A specific pattern of pain, posture, movement, muscle pathology, and reduced awareness of one's own body was found in women with CPP. These findings may increase our understanding of, and may point toward new treatment strategies for, this disease. PMID- 17084142 TI - Women's work stress and cortisol levels: a longitudinal study of the association between the psychosocial work environment and serum cortisol. AB - OBJECTIVE: The aim of the present study was to investigate whether there is an association between serum cortisol and work-related stress, as defined by the demand-control model in a longitudinal design. METHODS: One hundred ten women aged 47-53 years completed a health questionnaire, including the Swedish version of the Job Content Scale, and participated in a psychological interview at baseline and in a follow-up session 2 years later. Morning blood samples were drawn for analyses of cortisol. RESULTS: Multiple stepwise regression analyses and logistic regression analyses showed that work demands and lack of social support were significantly associated with cortisol. CONCLUSIONS: The results of this study showed that negative work characteristics in terms of high demands and low social support contributed significantly to the biological stress levels in middle-aged women. Participation in the study may have served as an intervention, increasing the women's awareness and thus improving their health profiles on follow-up. PMID- 17084143 TI - The role of work stress as a moderating variable in the chronic pain and depression association. AB - OBJECTIVE: This article aims to examine the role of work stress as a moderating variable in the chronic pain-depression association, as well as sex differences in this link. METHODS: The analyses were carried out using the Canadian Community Health Survey Cycle 1.1. Key variables were chronic pain conditions (fibromyalgia, arthritis/rheumatism, back problems, and migraine headaches), work stress, and depression. The total sample comprises 78,593 working individuals. RESULTS: In this working sample, 7.6% met criteria for major depression, but the prevalence increased to 12% in those also reporting chronic pain. Both depression and comorbid chronic pain and depression were twice as prevalent in women as in men. Having a chronic pain condition and overall work stress emerged as the strongest predictors of depression. Unexpectedly, however, none of the work stress domains moderated the chronic pain and depression association. CONCLUSION: The impact of work stress should be considered in the etiology and management of major depression. PMID- 17084144 TI - Chronic pain, depressive disorder, and the role of work. PMID- 17084145 TI - Posttraumatic stress disorder, tenderness, and fibromyalgia syndrome: are they different entities? AB - OBJECTIVES: Many features of fibromyalgia syndrome (FMS) resemble those of posttraumatic stress disorder (PTSD). The goal of this study was to investigate the comorbidity of FMS and PTSD in a cohort of men following an intensive, initial, defined traumatic event. METHODS: One hundred twenty-four males (55 patients with PTSD, 20 patients with major depression, and 49 controls) were evaluated for the presence of FMS. The major traumatic events in all PTSD patients were combat-related. Each individual completed questionnaires characterizing his disease, disabilities, and quality of life. RESULTS: Forty nine percent of PTSD patients, compared to 5% of major depression patients and none of normal controls, fulfilled the American College of Rheumatology criteria for FMS (P<.0001). Significant correlations were detected between tender points and measured parameters in the PTSD group. CONCLUSIONS: In male patients, PTSD is highly associated with FMS. The degree and impact of these disorders are also highly related. PMID- 17084146 TI - Psychophysiological responses in patients with fibromyalgia syndrome. AB - Physical and emotional stress and altered reactivity of the autonomic nervous system have been implicated in the development and maintenance of fibromyalgia syndrome (FMS). This study investigated blood pressure, heart rate (HR), skin conductance levels (SCL), and surface electromyograms (EMG) from the trapezius muscle in 30 FMS patients and 30 age- and sex-matched healthy controls (HCs). All measures were continuously recorded during baseline (BL), social conflict, mental arithmetic, and relaxation tasks. The FMS patients showed significantly higher stress ratings and self-reported stress responses. Baseline EMG levels were significantly lower, and BL HR was significantly elevated. During both stress tasks, HR reactivity was significantly lower, and SCL reactivity was significantly higher in the FMS group. This pattern of low BL muscle tension and high BL HR, along with low HR and high SCL reactivity to stress, is discrepant to other chronic pain syndromes and suggests unique psychophysiological features associated with FMS. Several potential mechanisms for these psychophysiological responses are discussed. PMID- 17084147 TI - Relationship between posttraumatic stress disorder symptoms and the course of whiplash complaints. AB - OBJECTIVE: This study investigates the relationship between posttraumatic stress disorder (PTSD) symptoms (avoidance, reexperiencing, and hyperarousal) and the presence, severity, and duration of neck complaints after motor vehicle accidents. METHODS: Individuals who had been involved in traffic accidents and had initiated compensation claim procedures with a Dutch insurance company were sent questionnaires (Q1) containing complaint-related questions and the Self Rating Scale for PTSD. Of the 997 questionnaires that were dispatched, 617 (62%) were returned. Only car accident victims were included in this study (n=240). Complaints were monitored using additional questionnaires that were administered 6 months (Q2) and 12 months (Q3) after the accident. RESULTS: PTSD was related to the presence and severity of concurrent post-whiplash syndrome. More specifically, the intensity of hyperarousal symptoms that were related to PTSD at Q1 was found to have predictive validity for the persistence and severity of post whiplash syndrome at 6 and 12 months follow-up. CONCLUSION: Results are consistent with the idea that PTSD hyperarousal symptoms have a detrimental influence on the recovery and severity of whiplash complaints following car accidents. PMID- 17084148 TI - Symptoms of PTSD according to individual and work environment characteristics of Korean railroad drivers with experience of person-under-train accidents. AB - OBJECTIVE: The purpose of this study is to collect basic data on the prevalence pattern of posttraumatic stress disorder (PTSD) in Korean railroad drivers exposed to person-under-train accidents according to the individual and working environmental characteristics. METHODS: Questionnaires were sent to all the railroad drivers accompanying active service now by company post. A total of 639 of the 800 questionnaires sent out were returned, showing a response rate of 79.9%. The Impact of Event Scale was utilized to address psychological status of railroad drivers after exposure to person-under-train accident. RESULTS: Score of PTSD, intrusion, and avoidance ranged from 9.6 to 13.2, from 4.2 to 6.1, and from 5.4 to 7.2 according to individual characteristics and ranged from 11.44 to 12.70, from 5.03 to 5.67, and from 6.41 to 7.04 according to working environmental characteristics, respectively. CONCLUSION: This study demonstrated that Korean railroad drivers who experienced person-under-train accident expressed adverse symptoms physically and psychologically more than those who did not experience it. Apart from effect of train driver's age, a wide range of individual and working environmental factors appear to have no association with the level of posttraumatic distress following a person-under-train accident. PMID- 17084149 TI - Positive and negative postdisaster psychological adjustment among adult survivors of the Southeast Asian earthquake-tsunami. AB - OBJECTIVE: This study explored psychosocial factors associated with psychological adjustment among survivors of a severe natural disaster. METHODS: Two hundred sixty-seven adult Thai survivors of the 2004 Southeast Asian earthquake-tsunami were surveyed. RESULTS: At 6 months following the disaster, percentages of survivors who reported positive and negative adjustment were 34% and 40%, respectively. For positive adjustment, best predictors were predisaster employment, increased arousal, and frequent support seeking. Support seeking also partially mediated the association between arousal and positive adjustment. For negative adjustment, infrequent support seeking and increased intrusion and arousal were the best predictors. Arousal was a partial mediator between intrusion and negative adjustment, and support seeking also partially mediated the influence of intrusion and arousal on negative adjustment. CONCLUSION: Mental health programs for survivors of natural disasters should focus on promoting active coping strategies such as support seeking, strengthening of support networks, and management of posttraumatic stress symptoms to facilitate adjustment. PMID- 17084150 TI - Psychological responses of pregnant women to an infectious outbreak: a case control study of the 2003 SARS outbreak in Hong Kong. AB - OBJECTIVE: The aim of the present study was to examine the behavioral and psychological responses of pregnant women during the 2003 severe acute respiratory syndrome (SARS) outbreak in Hong Kong. METHODS: Ethnographic interviews were first conducted to identify the common psychological and behavioral responses to the outbreak. This was followed by a case-control study of 235 consecutive pregnant women recruited during the SARS epidemic, and a historical cohort of 939 pregnant women recruited a year before the outbreak. Both cohorts completed standardized rating scales on depression, anxiety, and social support. RESULTS: Women in the SARS cohort adopted behavioral strategies to mitigate their risk of contracting infection. However, pregnant women tended to overestimate the risk of contracting SARS and nearly a third of the women were homebound. The anxiety level of the SARS cohort was slightly higher than that of the pre-SARS control. No statistical difference was found between the depression levels of the two cohorts. CONCLUSION: The improved social support experienced by pregnant women during SARS might have buffered the stress associated with an outbreak. However, clinicians should monitor for overestimation of infectious risk among pregnant women. PMID- 17084151 TI - Lower birth weight of Dutch neonates who were in utero at the time of the 9/11 attacks. AB - OBJECTIVE: Maternal stress during pregnancy has been reported to have an adverse influence on fetal growth. The terrorist attacks of September 11, 2001, on the United States have provoked feelings of insecurity and stress worldwide. Our aim was to test the hypothesis that maternal exposure to these acts of terrorism via the media had an unfavorable influence on mean birth weight in the Netherlands. METHODS: We compared birth weights of 1885 Dutch neonates who were in utero during the attacks with those of 1258 neonates who were in utero exactly 1 year later. RESULTS: In the exposed group, birth weight was lower than in the nonexposed group (difference, 48 g; 95% confidence interval=13.6, 82.9; P=.006). The difference in birth weight could not be explained by tobacco use, maternal age, parity, or other potential confounders or by shorter pregnancy durations. CONCLUSION: These results provide evidence supporting the hypothesis that exposure of Dutch pregnant women to the 9/11 events via the media has had an adverse effect on the birth weight of their offspring. PMID- 17084152 TI - Short-term psychological impact on family members of intensive care unit patients. AB - OBJECTIVE: This study aimed to evaluate the short-term psychological impact on family members of intensive care unit (ICU) patients during their stay in the unit. METHODS: Thirty-two first-degree relatives of patients treated in the ICU of two general hospitals were investigated for symptoms of early posttraumatic stress reaction, anxiety, and depression. Patients' and relatives' sociodemographic data and information pertaining to the patients' ICU treatment were collected. Family members were assessed at intake and before discharge through the Center for Epidemiological Studies Depression Scale, the State-Trait Anxiety Inventory, and the Impact of Event Scale. RESULTS: High rates of anxiety, depressive (97%), and posttraumatic stress (81%) symptoms were recorded at the initial assessment. Although symptoms remitted significantly, 87% and 59% of the sample fulfilled the criteria for a depressive and posttraumatic stress reaction, respectively, at the second assessment. Women exhibited higher levels of distress and more persisting symptoms than men did. Trait anxiety was the most significant predictor (P<.001) of the severity of depressive symptoms and the single predictor of the development of posttraumatic stress reaction (P<.000); also, state anxiety was a predictor of the development of posttraumatic stress symptoms, especially of the female gender. CONCLUSIONS: Family members of ICU patients exhibit high levels of distress that persist throughout their relatives' hospitalization. Women and individuals with high trait anxiety are at increased risk for developing such reactions. Severe early anxiety responses predicted the development of posttraumatic stress symptoms. Early case identification and intervention may prevent the full development of posttraumatic stress disorder. PMID- 17084153 TI - Denial of pregnancy as a reproductive dysfunction: a proposal for international classification systems. AB - OBJECTIVE: To propose a new Diagnostic and Statistical Manual (DSM) and International Classification of Diseases (ICD) category concerning reproductive dysfunctions not caused by organic disorder. METHODS: Forensic (n=93) and obstetrical cases (n=78) of denied and concealed pregnancies are compared and discussed. RESULTS: Denied pregnancies occur with a ratio of 1:475 births. An overwhelming phenomenological agreement is demonstrated between obstetrical and forensic cases. The findings were similar for denial and concealment of pregnancy, probably representing two different types of reproductive dysfunction that can be classified psychosocially together under the heading "negated pregnancy". CONCLUSIONS: Analogous to the phenomenology of "sexual dysfunctions", "reproductive dysfunctions" could be distinguished by the extraordinariness of both reproductive experience and behavior (as part of human sexuality) and psycho physiological changes, which characterize the reproductive course of events and cause obvious suffering and interpersonal complications. It is therefore suggested to include these entities as categories in their own right under the appropriate existing headings in DSM and ICD. PMID- 17084154 TI - Quality of life and depressive symptoms in patients with major depression and end stage renal disease: a matched-pair study. AB - OBJECTIVE: To compare the quality of life (QOL) and depressive symptoms of outpatients with major depression with that of nondepressed individuals undergoing hemodialysis (matched for age, gender, and ethnicity). METHODS: We used the WHOQOL BREF and the Beck Depression Inventory (BDI). RESULTS: After univariate analyses, depressed patients' QOL scores were significantly lower (P<.0001) in all the assessed domains (i.e., physical health, psychological, social relationships, environmental, and global QOL). Additionally, there was a significant difference in the severity of depression among the study groups (P<.0001). Finally, after multiple regression analysis, the QOL profile of depressed subjects remained significantly worse even when controlling for depression scores and diagnostic status. CONCLUSION: The subjective QOL of patients with major depression is significantly lower than that of subjects undergoing hemodialysis. PMID- 17084155 TI - Training model for special competence in general hospital psychiatry in Finland. PMID- 17084157 TI - Identifying men diagnosed with clinically localized prostate cancer who are at high risk for death from prostate cancer. AB - PURPOSE: We identified factors at diagnosis that are significantly associated with time to prostate cancer specific mortality following radical prostatectomy or external beam radiation therapy. MATERIALS AND METHODS: The study cohort included 1,453 men treated with radical prostatectomy (1,095) or external beam radiation therapy (358) for localized prostate cancer between 1989 and 2002. Cox regression multivariate analysis was used to evaluate whether prostate specific antigen, prostate specific antigen velocity, biopsy Gleason score and clinical tumor category at diagnosis were significantly associated with time to prostate cancer specific mortality following radical prostatectomy or external beam radiation therapy. RESULTS: In addition to increasing prostate specific antigen (p < or =0.04) and biopsy Gleason score 8 to 10 disease (p < or =0.02), prostate specific antigen velocity more than 2 ng/ml yearly was significantly associated with shorter time to prostate cancer specific mortality in patients treated with radical prostatectomy (adjusted HR 12, 95% CI 3 to 54) and external beam radiation therapy (adjusted HR 12, 95% CI 3 to 54) compared with that in men with prostate specific antigen velocity 2 ng/ml yearly or less (p < or =0.001). Despite low risk disease 7-year estimates of prostate cancer specific mortality were 5% to 19% in patients in whom prostate specific antigen increased by more than 2 ng/ml during the year before diagnosis compared with less than 1% in those with a prostate specific antigen increase of 2 ng/ml or less. CONCLUSIONS: Despite prostate specific antigen level less than 10 ng/ml and Gleason score 6 cancer a prostate specific antigen increase of more than 2 ng/ml during the year before diagnosis places a man at high risk for prostate cancer death following radical prostatectomy or external beam radiation therapy. PMID- 17084158 TI - Defining high risk prostate cancer with risk groups and nomograms: implications for designing clinical trials. AB - PURPOSE: Death from prostate cancer is usually preceded by metastases and it usually occurs in men with high risk disease who experienced biochemical failure with a short prostate specific antigen doubling time. We developed a model for determining disease specific survival in prostate cancer. MATERIALS AND METHODS: We used the model for defining high risk prostate cancer that was developed by the Radiation Therapy Oncology Group and combined it with the Kattan nomogram for predicting the risk of metastases. We selected 414 Radiation Therapy Oncology Group intermediate and high risk patients who were treated with external beam radiotherapy alone. Excluded were patients with low risk disease. The Kaplan Meier product limit method was used to estimate the probability of freedom from biochemical failure, overall survival and disease specific survival. RESULTS: A significant difference was observed in freedom from biochemical failure, disease specific survival and overall survival among the 3 tertiles created by the nomogram using the cutoff points less than 8.5%, 8.5% to 15% and greater than 15% (p <0.001, 0.0002 and 0.0003, respectively). Only the risk of metastases using the categorized nomogram score (less than 8.5% and 8.5% to 15% vs greater than 15%), not preradiotherapy prostate specific antigen or Radiation Therapy Oncology Group risk (Radiation Therapy Oncology Group 2 vs 3), was a significant predictor of disease specific and overall survival for intermediate/high risk patients and intermediate/high risk with 15% or less risk for metastases. CONCLUSIONS: We combined a risk group stratification scheme for disease specific survival with a nomogram predicting the risk of metastases and created a model that may be useful for designing phase III trials with metastases and disease specific survival as study end points. PMID- 17084160 TI - Defining high risk prostate cancer--where do we set the bar? A translational science approach to risk stratification. AB - PURPOSE: Risk stratification is commonly used in patients with prostate cancer but this effort has had no demonstrable effect on patient decision making for initial therapy. We propose new risk strata for clinically localized prostate cancer. MATERIALS AND METHODS: We examined current stratification methods for prostate cancer and their impact on prostate cancer therapy. RESULTS: Three risk strata for patients with clinically localized prostate cancer are proposed. Stratum 1 includes patients in whom active surveillance is associated with a low risk of disease progression. Stratum 2 includes patients in whom monotherapy, including external beam, interstitial radiotherapy or radical prostatectomy, is generally successful. Stratum 3 includes patients at high risk for recurrence with monotherapy in whom multimodal therapy may be superior. CONCLUSIONS: Risk stratification systems for prostate cancer should harmonize the needs of researchers to develop comparable groupings of patients, of patients who seek guidance on optimal therapy and of clinical trialists who seek to advance therapy for this disease. Our new stratification system provides such a structure. PMID- 17084162 TI - Initial therapy with radical prostatectomy for high risk localized prostate cancer. AB - PURPOSE: This study provides a perspective on initial treatment in select patients with high risk, localized prostate cancer. MATERIALS AND METHODS: A select literature review was done with commentary on the philosophy of initial surgery followed by adjuvant or salvage therapies. RESULTS: Early detection and associated stage migration identify a cadre of men with unfavorable but apparently localized prostate cancer who historically would not have been viewed as appropriate candidates for radical prostatectomy. Decreased morbidity from radical prostatectomy and data demonstrating improved outcomes in some patients treated with multimodal therapy protocols provide a rationale for including radical prostatectomy as part of an aggressive treatment plan to achieve optimal local elimination of cancer. Data suggest that radical prostatectomy and adjuvant or possibly even salvage radiation therapy may provide the best elimination of large local cancers. Whether such an approach provides results that are better than or even as good as those of the common standard of radiation therapy plus androgen deprivation therapy remains to be seen and, if so, at what cost to the patient in terms of adverse effects. However, it is likely that optimal elimination of local disease is needed to achieve the maximum benefit from adjuvant systemic endocrine, chemotherapy or targeted treatments. In other words optimal local therapy may be necessary but not sufficient. CONCLUSIONS: Initial radical prostatectomy may have a role for treating high risk localized prostate cancer. PMID- 17084163 TI - The case for adjuvant therapy for prostate cancer. AB - PURPOSE: The rationale for adjuvant therapy for prostate cancer was reviewed. MATERIALS AND METHODS: A selective review of the successful experience with adjuvant therapy in other epithelial neoplasms is presented along with discussion of the increasing success of chemotherapy to improve outcomes in advanced prostate cancer. RESULTS: Adjuvant therapy could be effective for prostate cancer if men with high risk features, such as Gleason score 8 or greater, prostate specific antigen more than 20 ng/ml, positive surgical margins with Gleason score 7 or greater, seminal vesicle invasion or positive lymph nodes, were selectively given additional therapy after primary surgery or radiation. These individuals are easily identified by routine pathological findings and precise recurrence risks can be calculated with available nomograms. For primary radiotherapy hormonal adjuvant treatment has now been shown to produce improved disease-free and overall survival. Adjuvant radiotherapy following prostatectomy decreases biochemical relapse and local disease recurrence. Adjuvant hormonal therapy after prostatectomy is also beneficial in select patients. Adjuvant chemotherapy now has a powerful rationale since it has been shown to improve survival in other common epithelial cancers, such as those of the breast, colon and lung. Newer agents produce significant response rates in patients with advanced metastatic prostate cancer, even prolonging overall survival. CONCLUSIONS: Only through dedication to a multimodality approach with all disciplines, including surgical urology, radiation therapy and medical oncology, will the promise of adjuvant therapy for prostate cancer be fully realized. PMID- 17084164 TI - Management of locally advanced prostate cancer: past, present, future. AB - PURPOSE: Historically advanced prostate cancer had been treated with androgen ablation. With the evolution of radiation therapy it was shown that some patients with advanced but nonmetastatic disease could be cured or at least have progression delayed. Subsequently a series of studies demonstrated that the combination of radiation and androgen ablation resulted in improved results over those of radiation therapy alone, although the failure rate was still high. This review explores the continued evolution in the treatment of high risk disease. MATERIALS AND METHODS: The published literature on treatment for high risk prostate cancer was reviewed. RESULTS: Adding androgen ablation to radiation decreased the failure rate from 79% to 67% in older studies and 55% to 25% in more recent studies. Most contemporary studies of higher radiation doses showed further improvement with a failure rate of 20% to 40%. The results of adding an implant boost appears to have decreased the failure rate further to 30% or less in most studies. There is now great interest in exploring chemotherapy or biological agents as adjuvant therapy to try to improve the results further. The role of surgery in these patients is also awaiting further clarification. CONCLUSIONS: Radiation therapy has been the primary mode of curative therapy for high risk prostate cancer for 3 decades. Much progress has been made. Evolving data suggest that radiation will continue to have the primary role in treatment in these patients in the future. PMID- 17084165 TI - The case for systemic therapy alone for prostate cancer. AB - PURPOSE: This review details clinical scenarios that may be appropriate for the use of primary androgen deprivation alone for organ confined prostate cancer. MATERIALS AND METHODS: The Medline and National Cancer Institute clinical trials databases were used. Searches were made regarding current and potential data to guide therapeutic decision making. RESULTS: Data integration that incorporates patient comorbidities and cancer histological features can identify patients at low vs high risk for death from prostate cancer. Observational databases have documented an increase in the use of primary androgen deprivation for low and high risk disease. Outcome data on androgen deprivation therapy alone in patients with metastatic as well as localized disease were reviewed. The potential of newer therapies, including chemotherapy, and therapies that target aberrant signaling pathways was also reviewed. The latter holds the potential to more effectively eradicate distant metastatic disease. CONCLUSIONS: Patients with high risk prostate cancer are those with a high chance of relapse with systemic disease despite treatment with definitive local therapy. Moreover, a patient with multiple comorbidities, and associated short life expectancy and high risk cancer may be a suitable candidate for systemic therapy alone with the goal of local and systemic disease control. In contrast, deferred systemic therapy alone until local progression and/or metastatic disease can be considered in a patient with low risk, indolent disease and a life expectancy of less than 10 years with the goal of avoiding over treating most patients, who often do not require any therapy during life. PMID- 17084166 TI - The case for androgen deprivation as primary therapy for early stage disease: results from J-CaP and CaPSURE. AB - PURPOSE: We analyzed the outcome of primary androgen depletion therapy, which has gained more attention as a potential therapeutic option in patients with localized or locally advanced prostate cancer as it has been increasingly implemented despite limited data on its therapeutic impact in Japan and the United States. MATERIALS AND METHODS: We analyzed data from CaPSURE and the Japanese Prostate Cancer study. RESULTS: In Japan primary androgen depletion therapy has long been the treatment of choice for localized and locally advanced prostate cancer. Based on CaPSURE data the frequency of primary androgen depletion therapy being chosen to treat localized and locally advanced disease is also increasing in clinical practice in the United States. A study of the outcomes of endocrine therapy is currently being performed in Japan by the Japanese Prostate Cancer Study Group. CONCLUSIONS: It is important to obtain such information about the role of primary androgen depletion therapy for localized and locally advanced prostate cancer from studies of natural history and clinical trials. It is also important to update practical treatment guidelines. PMID- 17084167 TI - Bone related events in high risk prostate cancer. AB - PURPOSE: We provide recommendations for defining and treating bone related events in high risk prostate cancer. MATERIALS AND METHODS: A focused literature review was done. RESULTS: Men with prostate cancer often have osteoporosis and osteopenia even before initiating androgen deprivation therapy. After starting androgen deprivation therapy they experience accelerated bone loss. Bone mineral density is the most common tool to assess the degree of bone loss, although the use of bone turnover markers for this purpose is being actively explored. Bisphosphonates are effective for increasing bone mineral density and treating osteoporosis. The benefits derived from bisphosphonates should be weighed against the adverse effects, including the risk of osteonecrosis of the jaw. Treatment is indicated in patients with prostate cancer with osteoporosis and it may be considered in patients with osteopenia and/or additional risk factors. The time of initiation of therapy and duration of treatment have not been conclusively established. CONCLUSIONS: Prolonged androgen deprivation therapy results in bone loss and it has a potential to impact quality of life. Additional research is needed to characterize patients who would benefit from therapy and optimize strategies to prevent osteoporosis. PMID- 17084169 TI - Management of high risk metastatic prostate cancer: defining risk at the time of initial treatment failure. AB - PURPOSE: Even as improvements in therapy success continue, initial treatment for prostate cancer fails in a substantial proportion of patients with apparently localized disease. However, the optimal approaches to defining biochemical failure and determining its clinical significance remain unclear. MATERIALS AND METHODS: A focused review and search of current literature were performed regarding biochemical recurrence and risk stratification after initial therapy for prostate cancer. RESULTS: Multiple definitions exist for prostate specific antigen failure after therapy for localized and metastatic prostate cancer. The timing of prostate specific antigen recurrence as well as prostate specific antigen kinetic characteristics, such as prostate specific antigen velocity and most significantly prostate specific antigen doubling time, impacts the prediction of posttreatment cancer specific and overall survival after biochemical recurrence is detected. In addition, as with determining the recurrence risk, Gleason score and pathological stage also can predict the survival likelihood. CONCLUSIONS: In most cases biochemical prostate specific antigen recurrence is the initial indicator of treatment failure and eventual progression but prostate specific antigen recurrence alone does not predict clinically significant events. As with determining the risk likelihood, pathological data and prostate specific antigen kinetic characteristics can help predict patient survival. PMID- 17084170 TI - Recommendations for defining and treating high risk localized prostate cancer. PMID- 17084171 TI - High risk biochemical relapse and the timing of androgen deprivation therapy. AB - PURPOSE: Although the timing of androgen deprivation therapy in patients with prostate specific antigen-only relapse remains controversial, a large amount of data exists supporting the use of androgen deprivation early in the clinical course in patients with high risk localized prostate cancer as well as in those with metastatic disease. We offer guidance on its use in patients with prostate specific antigen recurrence only. MATERIALS AND METHODS: We reviewed recent data on androgen deprivation for nonmetastatic prostate cancer, including updates from phase III, randomized studies of androgen deprivation in conjunction with radiation therapy or surgery in patients with high risk features. RESULTS: We commented on the extent to which these data on androgen deprivation for nonmetastatic prostate cancer could be extrapolated to the setting of recurrent disease. In addition, we reviewed retrospective analyses of androgen deprivation in patients with prostate specific antigen-only relapse and prognostic data regarding prostate specific antigen doubling time in recurrent disease. Furthermore, consideration was given to rapid prostate specific antigen doubling time as an indication for androgen deprivation. CONCLUSIONS: In patients with nonmetastatic prostate cancer with high risk features as well as in those with serological (prostate specific antigen-only) relapse androgen deprivation before the development of metastatic disease is indirectly supported by long-term outcomes in a series of clinical trials and retrospective data sets. Furthermore, prognostic data suggest that early androgen deprivation may be most beneficial in patients with serological relapse with prostate specific antigen doubling times less than 12 months. PMID- 17084172 TI - The case for secondary hormonal therapies in the chemotherapy age. AB - PURPOSE: Virtually all patients with high risk localized and metastatic prostate cancer who are treated with androgen deprivation therapy eventually have progressive clinical or biochemical disease despite this therapy. Despite this fact numerous therapies are available that target the interaction of androgen and androgen receptor in the castrate testosterone milieu and many clinical investigations are under way in this area. MATERIALS AND METHODS: This literature review focuses on the current clinical literature in support of secondary hormonal therapy. RESULTS: Despite low androgen levels the androgen receptor remains active through the amplification, mutation or alteration of coactivator function. These observations suggest that secondary hormonal therapies remain a reasonable clinical approach. Such approaches can be receptor or ligand directed. Receptor directed approaches to secondary hormonal therapy are antiandrogen withdrawal, sequential use of antiandrogens and estrogenic compounds. Ligand directed therapies are adrenal cortex inhibitors, such as ketoconazole and others in clinical development. Furthermore, in the context of androgen independent tumor growth in patients with metastatic disease clinicians are now faced with the choice of using chemotherapy or secondary hormonal manipulations. Appropriate patient selection is a critical component to the effective use of these agents. CONCLUSIONS: The modest activity of these secondary therapies challenges the notion that advancing prostate cancer uniformly becomes hormone refractory. It offers an alternative to the early use of chemotherapy in patients with androgen independent disease. PMID- 17084173 TI - The case for early chemotherapy for the treatment of metastatic disease. AB - PURPOSE: Several important questions are raised by the data from Southwest Oncology Group 99-16 and TAX 327. What is the optimal timing of chemotherapy for metastatic hormone refractory prostate cancer? Should asymptomatic patients be treated? Is there a theoretical survival advantage for treatment for hormone sensitive prostate cancer? We reviewed arguments for the early use of docetaxel in the hormone sensitive and hormone naive disease states. MATERIALS AND METHODS: Androgen independent prostate cancer was traditionally viewed as a chemoresistant disease. At best palliation of bone pain but not improved survival could be achieved with the combination of mitoxantrone and prednisone. Median survival rates for chemotherapy in this disease state were reported to be between 10 and 12 months. Phase II studies administering docetaxel weekly or every 3 weeks as a single agent or in combination with estramustine demonstrated median survival rates of 14 to 23 months, which appeared to be an improvement over standard therapy. Consequently the 2 randomized trials, Southwest Oncology Group 99-16 and TAX 327, were designed to confirm the preliminary observations of improved survival with docetaxel based therapy. RESULTS: TAX 327 and Southwest Oncology Group 99-16 treated asymptomatic as well as symptomatic patients. Thus, in contrast with mitoxantrone/prednisone, which was approved in symptomatic men with hormone refractory prostate cancer, the exact timing of the initiation of chemotherapy in docetaxel treated patients is still the subject of debate. CONCLUSIONS: Studies should be performed to investigate the optimal timing of chemotherapy as well as sequencing with androgen ablation in patients at high risk for progression and death from metastatic prostate cancer. Prognostic factors for death that have been identified in patients with androgen independent prostate cancer are hemoglobin, alkaline phosphatase, visceral disease and performance status. PMID- 17084174 TI - Management of high risk metastatic prostate cancer: the case for novel therapies. AB - PURPOSE: We reviewed the results of preliminary studies of select novel agents for metastatic prostate cancer and discuss the potential benefit of these agents for earlier stage disease, eg biochemically recurrent prostate cancer with high risk features. MATERIALS AND METHODS: Available data on select investigational immunotherapies as well as endothelin-A receptor antagonists and survivin inhibitors were obtained and reviewed through PubMed searches, conference proceedings and unpublished proprietary information, when available. RESULTS: A large number of promising agents are in varying stages of development. Phase III results have been reported for the endothelin-A receptor antagonist atrasentan. Several immunotherapies are currently in phase II/III trials, namely the GM-CSF transduced tumor cell vaccine GVAX, the prostatic acid phosphatase loaded dendritic cell vaccine Provenge and the prostate specific antigen expressing poxvirus vaccine PROSTVAC-VF. Another immunotherapy, the prostate specific membrane antigen immunoconjugate MLN2704 (Millennium Pharmaceuticals, Cambridge, Massachusetts), is in phase I/II study. The first clinical inhibitors of survivin are in early phase I studies. Several of these agents, including atrasentan, have shown statistically significant but modest effects in the advanced disease setting in which they have been studied. CONCLUSIONS: Clinical trial design with these novel therapies presents particular challenges since most of these agents may induce disease stabilization rather than disease regression. There is a risk of false-negative results and failure to recognize a potentially efficacious agent if these cytostatic agents are studied only in men with advanced, heavily pretreated disease in whom life expectancy is measured in months. We advocate the early referral and enrollment of men with high risk prostate cancer in clinical trials. PMID- 17084176 TI - Diastolic heart failure: perception of the syndrome and scope of the problem. AB - Diastolic Heart Failure: perception of the syndrome and scope of the problem. As a result of the recent publications of the CHARM-Preserved trial, the upcoming I PRESERVE and other still ongoing clinical trials in patients with Heart Failure and Normal (or Preserved) Left Ventricular Ejection Fraction, numerous questions have arisen as for the conceptual rationale and pathophysiological basis of such trials. The present symposium is a synopsis of the most important remaining controversies. We gratefully acknowledge all authors for their contribution as summarized in this Introduction Paper. PMID- 17084177 TI - Cardiac dysfunction in heart failure: the cardiologist's love affair with time. AB - Translating research into clinical practice has been a challenge throughout medical history. From the present review, it should be clear that this is particularly the case for heart failure. As a consequence, public awareness of this disease has been disillusionedly low, despite its prognosis being worse than that of most cancers and many other chronic diseases. We explore how over the past 150 years since Ludwig and Marey concepts about the evaluation of cardiac performance in patients with heart failure have emerged. From this historical physiologic perspective, we have seen how 3 increasingly reductionist approaches or schools of thought have evolved in parallel, that is, an input-output approach, a hemodynamic pump approach, and a muscular pump approach. Each one of these has provided complementary insights into the pathophysiology of heart failure and has resulted in measurements or derived indices, some of which still being in use in present-day cardiology. From the third, most reductionist muscular pump approach, we have learned that myocardial and ventricular relaxation properties as well as temporal and spatial nonuniformities have been largely overlooked in the 2 other, input-output and hemodynamic pump, approaches. A key message from the present review is that relaxation and nonuniformities can be fully understood only from within the time-space continuum of cardiac pumping. As cyclicity and rhythm are, in some way, the most basic aspects of cardiac function, considerations of time should dominate over any measurement of cardiac performance as a muscular pump. Any measurement that is blind for the arrow of cardiac time should therefore be interpreted with caution. We have seen how the escape from the time domain-as with the calculation of LV ejection fraction fascinating though as it may be, has undoubtedly served to hinder a rational scientific debate on the recent, so-called systolic-diastolic heart failure controversy. Lacking appreciation of early relaxation abnormalities and inappropriate degrees of nonuniformities has, indeed, led to some unfortunate misunderstandings about the pathophysiologic time progression of heart failure, in particular, heart failure with compensated hemodynamic pump function (ie, with normal or preserved LV ejection fraction). We have seen that with the introduction of newer powerful diagnostic techniques, as, for example, TDI and MRI, to evaluate ventricular "muscular pump" function, this debate can now be held in a more serene physiologic context. These aspects will be elaborated further in subsequent chapter papers of this symposium. With ongoing stem and other cell-based therapies and future reductionistic insights into cardiac cellular performance, we foresee the emergence of a fourth simple-parallel school of thought viewing the heart as a network of communicating different cell types, that is, cardiomyocytes, endothelial cells, fibroblasts, neurons. In this postgenomic age with the introduction of the rapidly evolving discipline of in vivo molecular imaging techniques, we anticipate that novel measurements of cardiac performance in patients with heart failure will soon become available and complement biopsy and other already available cardiac cellular biomarkers (cardiac troponin I; creatine kinase-MB; myoglobin; BNP). Through the use of these novel biomarkers as a fourth diagnostic track in the evaluation of cardiac performance in patients with heart failure, we will soon be able to increasingly understand the behavior of the heart as a complex biologic system-in other words, how these "low-level" biologic functions and signal transduction pathways at a cellular level contribute to the above "high-level" or system-level approach of cardiac performance at the muscular, the hemodynamic, and the input-output pump system levels and, hopefully, how they could contribute to an early diagnosis of chronic heart failure, in patients. PMID- 17084178 TI - Ventricular pump function in heart failure with normal ejection fraction: insights from pressure-volume measurements. AB - The syndrome of heart failure in the setting of normal ejection fraction (HFNEF) is manifest in a clinically heterogeneous group of patients with multiple and varied comorbid conditions. In this report, we review available data derived from pressure-volume (PV) analyses in patients with and in animal models of HFNEF. Pressure-volume analysis of ventricular function is challenging in the clinical setting but provides unique insights into the systolic, diastolic, and overall pumping characteristics of the heart. Results of such analyses have thus far been limited to small cohorts of patients but suggest that different cohorts of patients with HFNEF having PV relations that imply different pathophysiologic mechanisms exist. This emphasizes the need to take a view of this syndrome, which extends beyond diastolic dysfunction, particularly when it comes to proposing and investigating therapeutic targets. We therefore propose that progress can be made in advancing therapeutics for HFNEF if it is appreciated that different underlying pathophysiologic mechanisms may be important in different cohorts and if attention expands beyond diastolic dysfunction as the sole target. Similar to the success that was achieved in advancing therapeutics for systolic heart failure when attention shifted away from the heart to the neurohormonal and renal axes, our interpretation of data in human beings and in animal models suggests that addressing similar targets (perhaps not in exactly the same manner) may prove to be fruitful, at least for some patients with HFNEF as well. PMID- 17084179 TI - Systolic dysfunction in heart failure with a normal ejection fraction: echo Doppler measurements. AB - Heart failure with a normal ejection fraction, also called heart failure with preserved ejection fraction or diastolic heart failure, is thought to be characterized by normal systolic function and disturbed diastolic function only. However, studies using newer Doppler-echocardiographic techniques have shown that ventricular function is not normal particularly in the long axis. Ejection is relatively preserved because of increased radial function. Similar findings are seen with normal ageing and the typical precursors of heart failure with a normal ejection fraction such as hypertension, diabetes, and ischemia. There appears to be a spectrum of abnormalities of systolic function from the truly normal to systolic heart failure with heart failure with a normal ejection fraction occupying an intermediate position. The use of ejection fraction, which has a normal distribution, to dichotomize patients with heart failure is not supported on theoretical or experimental grounds, and any cutoff is arbitrary. Patients with heart failure have a mixture of systolic and diastolic abnormalities and variable degrees of remodeling. It is more important to correctly identify these in the individual patient. PMID- 17084180 TI - Systolic dysfunction in heart failure with normal ejection fraction: speckle tracking echocardiography. AB - Left ventricular (LV) ejection fraction (EF) and LV end-systolic volumes are important measures of LV global function. However, in many patients with symptoms of heart failure, these measures are within normal limits. This condition is classified as heart failure with normal ejection fraction (HFNEF). Reduced EF and increased end-systolic volume, however, require impaired function in a number of LV segments. Therefore, apparently normal systolic function in HFNEF may reflect limited sensitivity of global EF, and assessment of regional systolic function may provide important diagnostic information. The recently introduced method, speckle-tracking echocardiography (STE), represents a simplified, objective, and angle-independent modality for quantification of regional myocardial deformation. The software uses conventional gray-scale B-mode recordings and tracks myocardial speckles, which serve as natural acoustic markers. Radial and longitudinal myocardial deformation can be measured simultaneously from long-axis recordings, radial and circumferential deformation from short-axis recordings, and LV torsion from assessment of apical and basal short-axis rotation. Experimental and clinical studies have demonstrated that the STE method can assess myocardial function accurately in healthy subjects in the settings of acute and chronic ischemia, dyssynchrony, and cardiomyopathy. So far, no STE studies have been performed regarding HFNEF. The purpose of this article is therefore to illustrate the potential of this novel method. PMID- 17084181 TI - Cardiac dysfunction in heart failure with normal ejection fraction: MRI measurements. AB - Cardiovascular magnetic resonance is a non-invasive 3-dimensional imaging technique which can provide morphologic and functional information as well as tissue characterization without the use of ionizing radiation or nephrotoxic contrast agents. It has a high accuracy and reproducibility and is optimally suited to quantify structural and functional abnormalities and to follow a patient over time. In the setting of heart failure with normal ejection fraction it can be used as an alternative to echocardiography in those patients with suboptimal image quality but it can also provide unique information for the differential diagnosis and the underlying physiopathology of this syndrome. PMID- 17084184 TI - Comments about the prospective randomized trial by Aoyama et al. PMID- 17084185 TI - Radiosurgery plus or minus whole brain radiation therapy for the treatment of brain metastases. An editorial comment. PMID- 17084188 TI - A prospective evaluation of the role for intraoperative x-ray in lumbar discectomy. Predictors of incorrect level exposure. AB - BACKGROUND: Lumbar discectomy is among the most frequently performed procedures by spine surgeons. Among the potential difficulties encountered during this procedure, incorrect spinal level surgery remains a significant concern for surgeons and patients. Multiple groups have advocated the use of intraoperative x ray to reduce the incidence of incorrect level surgery; however, this technique has not been prospectively evaluated. METHODS: In an effort to determine the incidence of incorrect level exposure during lumbar discectomy and to define patient characteristics predictive of wrong level exposure, we examined 100 consecutive patients who underwent lumbar discectomy by a single surgeon. After exposure, the surgeon was asked to identify the level exposed, which was confirmed by intraoperative x-ray. Several patient characteristics were then examined by logistical regression to identify features predictive of a mismatch between level of exposure and level of pathology. RESULTS: The study population was composed of 48 men and 52 women who were aged 18 to 83 years. Patient weights ranged from 105 to 410 lb. There were 51 patients who had pathology at the L5-S1 level; 44 patients, L4-L5; 3 patients, L3-L4; and 1 patient, L2-L3. Four patients had transitional vertebrae. The intended level was initially exposed in 85% of cases. Age and level of pathology (P < .05) were identified as factors predictive of a mismatch between intraoperative level of exposure and preoperative level of pathology. CONCLUSIONS: Pathology above L5-S1 and patient age have been shown to reliably predict incorrect level exposure. Based upon the findings of this study, the routine use of intraoperative x-ray to confirm the level of exposure should be considered in all cases of lumbar discectomy. PMID- 17084186 TI - Controlled release of lipopolysaccharide in the subarachnoid space of rabbits induces chronic vasospasm in the absence of blood. AB - OBJECTIVE: Leukocyte-endothelial cell interactions appear to play a role in the development of vasospasm after SAH. Using a purely inflammatory protein, LPS, we evaluated the effect of inflammation on the development of chronic vasospasm in the absence of blood and compared it to SAH-induced vasospasm in rabbits. METHODS: Lipopolysaccharide was incorporated into EVAc polymers to produce 20% LPS/EVAc polymers (wt/wt). Rabbits (n = 23) were randomized to 4 experimental groups: (1) empty polymer (n = 6), (2) SAH (n = 5), (3) 0.7 mg/kg polymeric LPS dose (n = 6), and (4) 1.4 mg/kg polymeric LPS dose (n = 6). Blood and polymers were inserted into the cisterna magna. The rabbits were killed 3 days postoperatively, and the basilar arteries were harvested for morphometric analysis. Clinical response and lumen patencies were analyzed using ANOVA and a post hoc Newman-Keuls Multiple Comparisons test. RESULTS: Significant narrowing of the basilar artery was observed by insertion of 20% LPS/EVAc polymers into the subarachnoid space at a polymeric dose of 1.4 mg/kg (actual dose, 66 microg kg( 1) d(-1)) (75.4% +/- 4.2%; P < .01) and by SAH (80.3% +/- 8.1%; P < .01) as compared with the empty polymer group. A trend toward narrowing was observed in the 0.7 mg/kg polymeric LPS dose group (actual dose, 33 microg kg(-1) d(-1)) (85.2% +/- 2.6%; P > .05). Symptoms associated with SAH were noted in 50% of the rabbits in the 0.7 mg/kg LPS group and in 100% of rabbits in the 1.4 mg/kg LPS group. CONCLUSION: Controlled release of LPS into the subarachnoid space of rabbits produced chronic vasospasm in a dose-dependent manner. At a polymeric dose of 1.4 mg/kg, LPS-induced vasospasm was equivalent to that induced by SAH. This suggests that LPS and SAH may induce vasospasm through similar mechanisms and provides further evidence that inflammation plays a central role in the etiology of chronic vasospasm. PMID- 17084191 TI - Objective assessment of cervical spinal cord injury levels by transcranial magnetic motor-evoked potentials. AB - BACKGROUND: The neurologic examination serves as the optimal method to record the level of spinal cord injury (SCI). However, this test is subject to interexaminer variability. To address this shortcoming, we describe a technique that uses transcranial magnetic motor-evoked potentials (tcMMEPs) and dermatomal somatosensory-evoked potentials (d-SSEPs) to more accurately measure the precise level of SCI. METHODS: Two groups of subjects were studied: (1) complete cervical SCI (n = 10) and (2) neurologically intact volunteers (n = 10). Two additional patients were evaluated: one with a cervical central spinal cord syndrome and another with a head injury with a suspected cervical SCI. Each subject underwent upper extremity tcMMEPs and d-SSEPs. RESULTS: Transcranial magnetic motor-evoked potentials were elicited from all upper limb myotomes (C4-T1, bilaterally) in neurologically intact volunteers (20 sides). The level of injury was determined using tcMMEPs by observing the lowest level of measurable response. The level of injury obtained using tcMMEPs was the same as that determined by neurologic examination in 13 (65%) of the 20 sides. In 7 sides, tcMMEP responses were obtained 1 level lower than that assessed by physical examination. Dermatomal somatosensory-evoked potentials were obtained from all dermatomes of volunteers tested in the laboratory compared with only 5 of the 9 patients with SCI who underwent d-SSEP testing. CONCLUSION: Testing using tcMMEPs provides an objective supplement to the neurologic examination after acute cervical SCI. Dermatomal somatosensory-evoked potentials were of limited value in determining the level of cervical SCI. PMID- 17084194 TI - Stereotactic biopsy of brain stem masses: Decision analysis and literature review. AB - BACKGROUND: Adult brain stem tumors are rare, and diverse pathology can be found in this location. Stereotactic biopsy of lesions in the brain stem has been performed since the 1960s with high diagnostic and low complication rates. Advances in imaging technology have raised questions regarding the utility of biopsy. We perform decision analysis to aid clinicians in their approach to management of adult brain stem lesions. METHODS: A structured literature search revealed 20 publications with 457 patients who had undergone brain stem lesion biopsy. These publications were reviewed to determine diagnostic rates and the incidence of complications. Standard decision analytic techniques were applied to the case of a virtual adult patient with a lesion in the brain stem. RESULTS: A 1 way sensitivity analysis revealed the likelihood that the preoperative diagnosis was correct and the rate at which incorrect treatment was based on faulty empirical diagnosis as the 2 factors with the greatest effects on patient outcome. The diagnostic rate and complication rate of biopsy, within the ranges reported in the literature, had lesser effects. A threshold analysis was constructed to compare outcomes from stereotactic biopsy vs empiric therapy for a brain stem lesion. The probability that the preoperative diagnosis is correct is plotted vs the probability that empirical treatment based on an incorrect diagnosis will have adverse effect. CONCLUSIONS: Management of lesions in the adult brain stem requires careful consideration of multiple preoperative factors including clinical and radiographic diagnostic certainty, consequences of empiric therapy, and the surgeon's complication rate. PMID- 17084196 TI - Early surgical treatment vs conservative management for spontaneous supratentorial intracerebral hematomas: A prospective randomized study. AB - BACKGROUND: Treatment of primary SICH is still controversial. The aim of this study was to investigate the effectiveness of craniotomy and early hematoma evacuation vs nonoperative management in patients with SICH. METHODS: A prospective randomized study of craniotomy and early hematoma removal vs best medical management was performed in 108 patients with primary SICH. Surgical or medical treatment was initiated within 8 hours post ictus. Principal eligibility criterium was the presence of neurologic impairment associated with a spontaneous subcortical or putaminal hemorrhage bigger than 30 mL. Outcomes were assessed at 1 year post ictus. RESULTS: Analysis of outcome revealed a significantly higher percentage of GOS scores higher than 3 for the surgical patients, compared with those of the conservative group (33% and 9%, respectively; P < .05). By contrast, the mortality rates between operated and conservatively managed patients did not differ significantly. The main prognostic variables were the initial neurologic status, hematoma volume, and location. Stratifications of these parameters and analysis showed that the positive effect of surgery on the quality of survival was statistically not valid for patients with GCS scores lower than 8 or ICH volumes 80 mL or higher at the time of enrollment. CONCLUSIONS: The study demonstrates that surgical patients with subcortical or putaminal hematomas showed better functional results than their conservatively treated counterparts. However, early ICH evacuation failed to improve the survival rates, as compared with best medical management. PMID- 17084198 TI - Rising incidence of primary central nervous system lymphoma in Kumamoto, Japan. AB - BACKGROUND: Primary central nervous system lymphoma is an extranodal form of non Hodgkin lymphoma arising in the craniospinal axis. The incidence of PCNSL is reportedly on the increase in some parts of the world in the last 2 decades. METHODS: We surveyed 4007 patients diagnosed with PIT between 1989 and 2004, with histologic diagnosis being obtained in 70% of the patients. Of these, the PCNSL cases were reviewed. RESULTS: Of 4007, 136 (3.4%) carried a diagnosis of PCNSL. At 0.41 per 100,000 per year, the age-adjusted incidence rate for that period was higher than that for the period from 1989 to 1998 (0.29 per 100,000 per year). Moreover, the number of patients with PCNSL doubled from 45 (1989-1996) to 91 (1997-2004). CONCLUSIONS: Our findings point to an increase in the incidence of PCNSL among immunocompetent individuals in Kumamoto Prefecture. PMID- 17084199 TI - Long-term clinical and radiologic results of small cerebral aneurysms embolized with 1 or 2 detachable coils. AB - BACKGROUND: Small cerebral aneurysms embolized with only 1 or 2 detachable coils often seem unstable and unsatisfactory. We tried to assess the long-term results of such embolized aneurysms. METHODS: Among 231 intracranial saccular aneurysms embolized at the Asan Medical Center between July 1995 and July 2004, 27 small aneurysms were occluded with only 1 (n = 12) or 2 (n = 15) detachable coils. Clinical and radiologic results were evaluated retrospectively for the involved patients. RESULTS: For the 1- and 2-coiled aneurysms, respectively, the mean dome sizes were 3.0 and 4.6 mm and the mean neck sizes were 1.9 and 2.7 mm. Most of the aneurysms were in the posterior circulation (19/27, 70%) and were found after bleeding (17/27, 63%). At the end of the embolization, complete occlusion in 19 aneurysms, residual neck in 1 aneurysm, and residual aneurysm in 7 aneurysms were attained angiographically. The mean packing ratios were 17.9% and 20.7% for the 1 and 2-coiled aneurysms, respectively. The mean clinical follow-up period of the patients was 41 months. No patient showed evidence of rebleeding; in addition, 1 patient with 1 coil and 2 patients with 2 coils were not followed up after discharge. Among the 10 follow-up DSAs or MRAs acquired after more than 8 months, radiologic major recurrences were detected in 2 patients after 10 and 15 months and second embolizations were done. CONCLUSIONS: Although the 1- and 2-coiled small aneurysms with a relatively low packing ratio seemed unstable, they showed a relatively low incidence of rebleeding and recurrence. PMID- 17084200 TI - Percutaneous angioplasty of a chronic total occlusion of the intracranial internal carotid artery. Case report. AB - BACKGROUND: A CTO of the intracranial ICA is usually managed medically and rarely by EC-IC bypass in selected patients. Percutaneous transluminal angioplasty has not been used. CASE DESCRIPTION: A 73-year-old man presented with frequent temporary blindness of the left eye and dizziness due to thrombotic occlusion of the left intracranial ICA, causing hemodynamic compromise. This patient was successfully treated by percutaneous angioplasty (balloon angioplasty and stent placement) under proximal balloon protection at 7 weeks from the ictus. Ischemic symptoms had not recurred during the 6-month follow-up period. CONCLUSION: Percutaneous angioplasty for a CTO of the intracranial ICA is technically feasible and can be an alternative to EC-IC bypass in a selected group of patients with symptomatic hemodynamic compromise, which is refractory to the best medical treatment. PMID- 17084201 TI - Tentorial dural arteriovenous malformation manifesting as trigeminal neuralgia treated by stereotactic radiosurgery: A case report. AB - BACKGROUND: Tentorial dAVMs as a cause of trigeminal neuralgia are extremely rare. Consensus exists that radical treatment of such high-flow dAVMs should be considered because of their high risk of bleeding. The authors present a case of a 50-year-old man who presented with symptoms of tic douloureux caused by tentorial dAVM, which was treated successfully by SRS. CASE DESCRIPTION: The patient consulted us with a 1-month history of harboring right trigeminal neuralgia. The cerebral angiography revealed a dAVM in the petrotentorial region and the MRI demonstrated a dilated petrosal venous varix compressing the root entry zone of the right trigeminal nerve. Gamma knife surgery was performed with a marginal dose of 18 Gy and a maximum dose of 30 Gy in a volume of 0.3 mL. The target point of the radiosurgery for this patient was a fistula identified by superselective angiography, enhanced computed tomography, and MRI. Follow-up MRI, 1 year after GKS, showed a thrombosed lesion, and the patient was able to end medication. There was no evidence of recurrence or adverse effects in the 3-year follow-up. CONCLUSION: Stereotactic radiosurgery can play an effective role as a treatment modality for such unusual dAVMs. PMID- 17084203 TI - Venous loop mimicking middle cerebral artery bifurcation aneurysm on computed tomographic angiography--case report. AB - BACKGROUND: CT angiography has been widely used for the detection of cerebral aneurysm. However, there are still limitations despite improving CT angiographic techniques. We describe the unusual case of the detection of a false-positive aneurysm on CT angiography. CASE DESCRIPTION: A 64-year-old man presented with a 14-day history of severe headache and nuchal rigidity. Brain CT demonstrated no subarachnoid hemorrhage, but cerebrospinal fluid appeared xanthochromic in 3 successive tubes after lumbar puncture. Subsequent CT angiography suggested an aneurysm at the bifurcation of the left MCA. However, at operation, prominent sylvian vein superimposed on the bifurcation of the left MCA, and there was no aneurysm. Follow-up monitoring with conventional angiography at 2 weeks showed no abnormal findings. The patient recovered uneventfully. CONCLUSION: The case demonstrates that although CT angiography has a reportedly high specificity and sensitivity for the detection of intracranial aneurysm, careful interpretation is required to diagnose intracranial aneurysms. PMID- 17084206 TI - Benign osteoblastoma of the temporal bone: Case report and review of the literature. AB - BACKGROUND: Benign osteoblastoma arising from the temporal bone is extremely rare in elderly patients. We reviewed the literature on benign osteoblastoma of the temporal bone and now propose a new classification of this pathologic entity based on its anatomical location in the temporal bone. CASE DESCRIPTION: A 68 year-old woman presented with tinnitus and hearing disturbance with a duration of 1 year. Her neurologic examination revealed mixed hearing disturbance and hypogeusia. Preoperative computed tomography showed a bony destructive isodensity mass with calcified component involving the temporal bone. The FDG-PET scan showed high uptake on the lesion. After preoperative embolization on the day before surgery, the patient underwent tumor removal via a left infratemporal approach under neuronavigated guidance. The pathologic examination revealed a benign osteoblastoma. CONCLUSION: We present the first case of FDG-PET showing high uptake on the lesion. Preoperative embolization is useful to reduce the amount of bleeding during the surgery. PMID- 17084204 TI - Late development of intraventricular papillary pituitary carcinoma after irradiation of prolactinoma. AB - BACKGROUND: Intracranial dissemination of pituitary adenomas is a rare event that does not equate malignancy. Most of the cases have been reported as metastases from pituitary carcinoma. A case of papillary pituitary carcinoma developed 12 years after radiotherapy for prolactin-secreting hormone pituitary adenoma is presented. CASE DESCRIPTION: A 37-year-old woman was admitted for the second time with neurologic disturbance and hypertension. A gadolinium-enhanced magnetic resonance scan of the brain demonstrated a 50-mm enhanced mass (absent on previous studies) on the lateral ventricle, involving the left temporal lobe. The patient underwent a craniotomy and biopsy of the lesion that was consistent with pituitary carcinoma; it was immunoreactive to follicle-stimulating hormone, adrenocorticotropic hormone, pituitary tumor-transforming gene, and epithelial cell adhesion molecule. Transmission electron microscopy analysis confirmed the secretory pituitary tumor diagnosis. CONCLUSIONS: The tumor was considered a primary pituitary papillary carcinoma. The clinical course indicated that this tumor was the seedling of a pituitary tumor, although it could be interpreted as metastases from a pituitary carcinoma. PMID- 17084207 TI - Primary malignant B-cell-type dural lymphoma: Case report. AB - BACKGROUND: The primary malignant dural lymphoma of B-cell type is rare. Our review of the literature (24 cases) indicated that patients with this tumor had female predominance, immunocompetency, and longer survival compared with those with primary malignant intracerebral lymphomas. Based on its clinicopathological features, this dural lymphoma may be classified differently from other types of malignant lymphomas in the central nervous system. CASE DESCRIPTION: The authors report an example of a patient who had a favorable course of malignant dural lymphoma. A 59-year-old woman presented with primary malignant dural lymphoma in the frontal area. Histological diagnosis was a diffuse large B-cell-type lymphoma of intermediate malignancy, but MIB-1 index was extremely high. Subtotal tumor removal was followed by 4 courses of chemotherapy. Thirty months after surgery, the patient is asymptomatic with no MRI evidence of recurrence. A combination of subtotal resection and chemotherapy for a primary malignant dural lymphoma is an effective means to attempt cure of this tumor. CONCLUSION: These rare tumors may be managed successfully by subtotal resection and postoperative treatment. We believe that chemotherapy is the treatment of choice after subtotal resection of malignant dural lymphomas, as experienced in our case. PMID- 17084209 TI - Transcavum septum pellucidum interforniceal approach for the colloid cyst of the third ventricle Operative nuance. AB - BACKGROUND: Despite their unfavorable locations, lesions of the third ventricle can be successfully removed via an interhemispheric, transcallosal approach. In cases with normal ventricular anatomy, this approach requires unilateral or bilateral identification of the foramen of Monro. TECHNIQUE: However, in the presence of abnormal ventricular configuration such as cavum septum pellucidum (CSP), this basic knowledge needs to be modified. After routine callosotomy, there may be a confusion while entering the CSP due to the invisualization of ventricular landmarks such as the foramen of Monro, thalamostriate vein, and choroid plexus. The floor of the CSP is formed by the fornices, and a direct approach to the interforniceal area is easier via the CSP. But the interforniceal approach is not a routine way to reach the third ventricle, which has higher risks than other modalities. CONCLUSION: This approach should be planned and used in selected cases of the CSP. Opening of the walls of CSP is recommended both to expose both the foramen of Monro and to gain safe access to the third ventricle before manipulating the interforniceal area. PMID- 17084212 TI - Leadership vs. consensus. PMID- 17084213 TI - Re: Subarachnoid hemorrhage on computed tomography scanning and the development of cerebral vasospasm: the Fisher grade revisited (Smith ML et al. Surg Neurol 2005;63:229-35). PMID- 17084214 TI - Medtronic "fusiongate". PMID- 17084215 TI - Re: Why omega-3 fatty acids are important to neurosurgeons (Ausman JI, Surg Neurol 2006;65:325). Omega-3 fatty acids (fish oil) as an anti-inflammatory: an alternative to NSAIDs for discogenic pain (Maroon JC and Bost JW, Surg Neurol 2006;65:326-31). PMID- 17084218 TI - All along the CHD/CVD risk watchtower. PMID- 17084217 TI - Rhinosinusitis: developing guidance for clinical trials. AB - The Rhinosinusitis Initiative was developed by 5 national societies. The current guidance document is an expansion of the 2004 publication "Rhinosinusitis: Establishing definitions for clinical research and patient care" and provides templates for clinical trials in antimicrobial, anti-inflammatory, and symptom relieving therapies for the following: (1) acute presumed bacterial rhinosinusitis, (2) chronic rhinosinusitis (CRS) without nasal polyps, (3) CRS with nasal polyps, and (4) classic allergic fungal rhinosinusitis. In addition to the templates for clinical trials and proposed study designs, the Rhinosinusitis Initiative has developed 6 appendices, which address (1) health outcomes, (2) nasal endoscopy and staging of CRS, (3) radiologic imaging, (4) microbiology, (5) laboratory measures, and (6) biostatistical methods. PMID- 17084219 TI - Foreign bodies in the aerodigestive tract--a clinical study of cases in the coastal belt of South India. AB - PURPOSE: To investigate the cases of foreign body in the aerodigestive tract and to elucidate the characteristic problems in India. DESIGN AND SETTING: Three hundred patients presenting with a history of either aspirating or swallowing a foreign body were analyzed in a tertiary care center. Data were collected by retrospective chart review and statistically analyzed. RESULTS: Of the 300 patients studied, 90% (270 cases) did have a retained foreign body, with 233 (86.2%) cases found in the pharyngoesophagus and 37 (13.7%) cases in the tracheobronchial tree. With foreign bodies in the tracheobronchial passages, the most common sign and symptom were rhonchi (50%) and dyspnea (67%), respectively; in pharyngoesophagus, the most common symptom and sign were dysphagia (64%) and odynophagia (45%), respectively. The most common foreign body found was fish bone in 39% (106 cases) and the most common site of lodgment was the cervical esophagus 50.5% (136 cases). Rigid endoscopy with forceps removal under general anesthesia was the main treatment modality. Thirty-three (12.2%) cases had complications secondary to retained foreign body. CONCLUSION: Foreign bodies in the aerodigestive tract continue to be a common problem affecting adults and children alike. Rigid endoscopies with forceps removal under general anesthesia are the preferred management modality. From their experience, the authors recommend that no foreign body in the upper aerodigestive tract should be left alone with the hope that it will come out spontaneously. Delay in diagnosis and management can lead to life-threatening complications. PMID- 17084220 TI - Long-term voice and swallowing modifications after supracricoid laryngectomy: objective, subjective, and self-assessment data. AB - PURPOSE: The supracricoid laryngectomies (SCLs) are conservative surgical techniques for the treatment of selected laryngeal carcinomas. The advantage of SCL is that a permanent tracheostoma is not required, thus, maintaining the principal laryngeal functions. The aim of the study is to report objective, subjective, and self-assessment long-term results of voice and swallowing in a large group of patients who underwent SCL at least 2 years before this study was undertaken. METHODS: Twenty male subjects who underwent SCL with a mean age of 71 years (range, 51-82) were involved in a retrospective study on swallowing and vocal function. Videoendoscopic ratings were taken of neoglottic vibration patterns and bolus transit. The maximum phonation time and the syllable diadochokinesis were measured. Spectrograms were recorded. Voices were perceptually rated using the Grade, Instability, Roughness, Breathiness, Asthenicity, Strain (GIRBAS) scale. All of the subjects completed a self assessment questionnaire for both voice and swallowing. RESULTS: Videoendoscopic ratings showed moderate to severe impairment of neoglottal vibration, whereas bolus transit appeared only mildly impaired. The mean values of the GIRBAS scale were 2.4, 2.6, 2.4, 0.8, 0.5, and 0.8. The mean maximum phonation time was 7.5 seconds, whereas the mean value of the Yanagihara scale was 3.7 for voice spectrograms. Mean syllable diadochokinesis appeared as 3.3 syllables per second. Voice and swallowing quality of life questionnaires revealed satisfied patients. CONCLUSIONS: Swallowing after SCL was satisfactory; on the contrary, endoscopic, aerodynamic, perceptual, and acoustic data showed a highly dysphonic voice after SCL. However, self-assessment results revealed relatively satisfied speakers on the emotional, physical, and functional levels. PMID- 17084221 TI - Management of refractory chronic rhinosinusitis in children. AB - OBJECTIVE: Pediatric chronic rhinosinusitis (CRS) continues to be a difficult problem for the medical community. Traditionally, oral antibiotic therapy has been the cornerstone of treatment, but some patients, who are resistant to multiple trials of oral antibiotics, require alternative therapeutic modalities. One such option, the use of intravenous (IV) antibiotics, has shown a high success rate in a limited study. Our goal is to estimate the success of a therapy consisting of culture directed IV antibiotics, adenoidectomy, and sinus aspiration in ameliorating long-term (>12 months) clinical symptoms of CRS in pediatric patients who are resistant to oral antibiotic therapy. METHODS: We conducted a retrospective review of the medical records of 22 patients who received treatment, focusing on patient age, initial CRS presenting symptoms, computed tomography scan finding, length, duration, and type of prior oral antibiotic treatment, length and type of IV antibiotic treatment, and long-term follow-up of clinical symptom resolution. RESULTS: Initial clinical improvement after cessation of IV therapy was achieved in all 22 (100%) patients. Seventeen (77%) of the patients demonstrated long-term resolution of clinical symptoms of CRS. Excluding 4 immunocompromised patients, 16 of 18 (89%) of patients demonstrated long-term amelioration of CRS symptoms. CONCLUSIONS: The results suggest that IV antibiotics may prove beneficial for pediatric patients who demonstrate CRS not responsive to traditional oral therapy. Although other treatment options are currently available for this subpopulation of patients, this IV antibiotic therapy provides both a maximally effective outcome while using a relatively minimally invasive intervention. PMID- 17084222 TI - Otogenic intracranial complications. a 7-year retrospective review. AB - OBJECTIVES: The objectives of this study were to review our experience on intracranial complications secondary to otitis media (OM), and to compare the results to those occurring 10-15 years ago. We also reviewed the timing of both otologic and neurosurgical treatments. MATERIAL AND METHODS: All patients with intracranial complications secondary to OM over a 7-year period were identified. A retrospective chart review was undertaken at Beaumont University Hospital, Dublin, Ireland. Clinical presentation, radiological findings, microbiology, surgical management, and antibiotic use were studied. We compare our findings to those of other international investigators. RESULTS: Twelve cases were identified. Five had brain abscesses, 4 had lateral sinus thrombosis, and 3 had petrous apicitis. Eight of these cases were secondary to chronic OM and 4 were secondary to acute OM. Malodorous otorrhea, otalgia, headache, fever, and vertigo were the primary symptoms. Anaerobic bacteria were the most commonly isolated organisms followed by Staphylococcus aureus, Proteus mirabilis, and Pseudomonas aeruginosa. Patients with cholesteatoma underwent modified radical or radical mastoidectomy, and those who did not have cholesteatoma underwent cortical mastoidectomy. Two brain abscesses were drained before mastoid surgery; 2 were drained after mastoid surgery and 1 at the same time as otologic surgery. All patients received broad-spectrum intravenous antibiotics targeted at individual culture and sensitivity results. The mortality rate was zero. CONCLUSION: Clinical presentation and the frequency of occurrence of intracranial complications are similar to those occurring 10 years ago. The choice of antibiotics should include adequate anaerobic cover. We recommend that otologic surgery be performed at the same time as intracranial surgery for patients with mature brain abscesses. PMID- 17084223 TI - The preauricular sinus: factors contributing to recurrence after surgery. AB - PURPOSE: The objective of this study was to summarize clinical presentation, treatment, and recurrence of preauricular sinuses. MATERIALS AND METHODS: This retrospective, institutional review board-approved study reviewed the medical records of patients who underwent preauricular fistulectomy between January 1995 and June 2005 at university-based hospitals in South Korea. Only patients who underwent classic preauricular fistulectomy (not incision and drainage) and could be followed up for at least 3 months were included in the study. RESULTS: A total of 191 patients (206 ears) were enrolled. The right and left ears were involved in 79 and 97 patients, respectively. The most common location of the preauricular pit was the anterior margin of the ascending limb of the helix (93.2%). The most common indication for surgery was the recurrent exacerbation of acute infection (58.3%). The recurrence rate after surgery was 4.9%. Surgery under local anesthesia contributed to recurrence after the procedure (P = .009) and the cases that featured local infiltrative anesthesia had a higher rate of recurrence than the cases that had general anesthesia with an odds ratio of 6.875. CONCLUSIONS: Although this study showed that surgery under local anesthesia contributed to recurrence, it did not mean that it was only the anesthesia technique that influenced the recurrence. Surgeons should bear in mind that complete removal of the epithelial lining provides a lower recurrence rate, especially under local anesthesia. The main limitation of this study was that recurrent cases were too few to allow a statistical analysis. PMID- 17084224 TI - Modification of lymphocyte subsets in patients with rhinoscleroma. AB - PURPOSE: Rhinoscleroma is a rare, chronic, granulomatous disorder of the upper airways. This disease presents some etiopathogenetic aspects that are not yet clear. Infection by Klebsiella rhinoscleromatis is fundamental for the onset of the disease, but it is impossible to reproduce rhinoscleroma experimentally only via infection with the bacteria both in man and in animals. Furthermore, this disease mainly affects blood-related people and occurs in certain geographic areas. In this context, we present a study that brings to light some of the quantitative abnormalities of the lymphocyte subsets. MATERIALS AND METHODS: The study group consisted of 5 patients with rhinoscleroma. The following parameters were studied for each patient: clinical manifestations, histologic examinations, number of leukocytes, lymphocytes, and lymphocyte subsets. RESULTS: In all patients, we noted the following: There was a relative reduction of the CD4+ cells, an absolute increase of the CD8+ cells, and an inversion of the CD4+/CD8+ ratio. There was an absolute increase of the CD56+ cells and cytotoxic cells that coexpress CD8+CD56+ antigens. There was a relative reduction of the CD3+ cells, and the CD19+ cells tended to show an ambiguous behavioral pattern. CONCLUSION: We believe that K. rhinoscleromatis does not play a major role in the etiopathogenesis of rhinoscleroma. However, we do believe that the anomalous behavior of the immune system can favor rhinoscleroma. PMID- 17084225 TI - Sphenoid sinus chondromyxoid fibroma mimicking a mucocele. AB - We report the case of a 44-year-old man who presented with a chondromyxoid fibroma (CMF) of the sphenoid sinus, which filled the sinus and clinically and radiologically resembled a mucocele. Chondromyxoid fibromas are the least common cartilaginous neoplasms of bone, typically occurring in the metaphysis of long bones. They have occasionally been described in the facial bones, usually the mandible and maxilla, and rarely involve the paranasal sinuses. Chondromyxoid fibroma displays a relatively unique histological appearance, with characteristic lobulation, with spindle to stellate cells embedded in a myxoid or chondroid matrix. The periphery of the lobules tends to be more cellular than the center. Despite having been described nearly 60 years ago, the histogenesis of CMF remains controversial. Although CMFs are generally regarded as benign neoplasms, they may show an infiltrative pattern and may recur, particularly when they are in locations where complete surgical excision may be difficult or impossible. PMID- 17084226 TI - Recurrent bilateral dacryocystoceles in Wegener's granulomatosis: a rhinologic perspective. AB - Wegener's granulomatosis (WG) is a rare, idiopathic, systemic vasculitis of small vessels that manifests in multiple organ systems. Otorhinolaryngic manifestations of this disease include recurrent sinusitis and relapsing polychondritis. Periocular involvement is also a well-documented location of Wegener's disease. We present the case of a 13-year-old girl with severe WG who developed multiple recurrent orbital infections. She underwent multiple incision and drainage surgeries of each orbit and multiple courses of intravenous antibiotics. The patient persistently reaccumulated purulence in her nasolacrimal duct system and was referred to an oculoplastic surgeon for evaluation of these recurrent infections. The diagnosis of dacryocystitis as a complication of WG was made. This unique case represents a patient with severe WG developing bilateral dacryocystitis requiring bilateral dacryocystorhinostomies. PMID- 17084227 TI - Safe extraction of an impacted open safety pin from the esophagus: report of 9 cases. AB - PURPOSE: Foreign body ingestion is a common problem in children. An infrequent ingested item is an open safety pin (SP), which may become lodged in the esophagus and presents the endoscopist a challenge to his surgical judgment and technical skills. MATERIALS AND METHODS: We present 9 cases of infants with open SPs impacted in the esophagus and discuss the different options to treat these cases. RESULTS AND CONCLUSIONS: The management of open SP ingestion is discussed, along with a review of the literature regarding this rare condition. PMID- 17084228 TI - Penetrating vestibular injury due to a twig entering via the external auditory meatus. AB - We report a patient in whom a twig entered the vestibule via the external auditory meatus and was removed surgically. The patient was an 11-year-old boy who sustained a penetrating injury to his ear when he was rock climbing. Vertigo and hearing loss developed immediately. The vertigo improved over time. As it recurred, however, exploratory tympanotomy was performed 17 days after the accident. A piece of wood about 8 mm long was found to have penetrated the vestibule from in front of the stapes. This foreign body was removed and the vestibular fistula was closed. Although the patient's vertigo resolved after the operation, his hearing did not improve. PMID- 17084229 TI - Congenital transsphenoidal meningocele: case report and review of the literature. AB - Intrasphenoidal meningoencephalocele is a rare clinical entity. Its origin can be congenital, traumatic, tumoral, or spontaneous. Presenting as cerebrospinal fluid fistula with rhinorrhea, the diagnosis and treatment of this uncommon disease is a real challenge for the otorhinolaryngologist. We report a case of sphenoidal meningocele treated using an endoscopic procedure and review the literature regarding its congenital origin. PMID- 17084230 TI - Esophageal regurgitation as a cause of inspiratory distress after thyroplasty. AB - OBJECTIVES: Edema of the arytenoid and wound hematoma are principal causes of inspiratory distress after arytenoid adduction and type I thyroplasty. The purpose of the present study was demonstrate esophageal regurgitation one of the causes of inspiratory distress after thyroplastic surgeries. STUDY DESIGN: Two case reports. METHODS: We encountered 2 patients with unilateral vocal fold immobility who required emergent tracheostomy owing to sudden inspiratory distress 1 to 2 days after completion of arytenoid adduction combined with type I thyroplasty. RESULTS: Their dyspneic attacks occurred just after taking a meal. Both had a history of esophagectomy with reconstruction of the food passage using a gastric tube. They had a sufficiently wide glottis as indicated by laryngeal fiberscopy. Videofluorography showed a stricture at the junction between the duodenum and the gastric tube and barium pooling above the stricture. Regurgitation of barium was also seen. CONCLUSION: Based on these clinical courses and findings, laryngeal closure reflex triggered by esophageal regurgitation was considered to be the most possible cause of their dyspneic attacks. Phonosurgeons considering thyroplastic surgeries for postesophagectomy patients should be aware that esophageal regurgitation possibly causes laryngeal closure reflex resulting in inspiratory distress during the postoperative period. PMID- 17084231 TI - Tricholemmoma of the nose. AB - OBJECTIVES: The objectives of this case report are to (1) identify clinical presentations of tricholemmoma, (2) discuss the characteristics of Cowden syndrome and the relationship between this syndrome and tricholemmoma, and (3) differentiate tricholemmoma from other superficial cutaneous tumors such as basal cell carcinoma and granulomas. STUDY DESIGN: We present a literature review and a case report. METHODS: A literature review of tricholemmoma and a case report of an unusual manifestation of this neoplasm were presented. RESULTS: Tricholemmoma was first described as a cutaneous neoplasm in 1962. It is associated with the Cowden syndrome and can be misdiagnosed as a more aggressive cutaneous malignancy. We report an unusual case of tricholemmoma presenting as a nasal mass. CONCLUSIONS: Tricholemmoma is described as having a predilection for the head and neck, yet little information has been published about this disorder within the Otolaryngology literature. Otolaryngologist-head and neck surgeons must be familiar with this neoplasm because it can be frequently misdiagnosed as an aggressive cutaneous malignancy and hence can be incorrectly treated. We describe the presentation and diagnosis of tricholemmoma, describe the frequent association of this neoplasm with Cowden syndrome, and report an unusual presentation of tricholemmoma as an intranasal mass. PMID- 17084232 TI - HIV infection initially presenting as sinonasal Burkitt's lymphoma. AB - Burkitt's lymphoma is a common comorbidity of infection with the human immunodeficiency virus, but rarely the initial clinical manifestation of undiagnosed AIDS, especially for the otolaryngologist. We report the case of a patient with recalcitrant rhinosinusitis subsequently diagnosed with sinonasal Burkitt's lymphoma and HIV. PMID- 17084233 TI - Mandibular distraction osteogenesis to relieve Pierre Robin airway obstruction. AB - Children with Pierre Robin sequence can present challenging airway management issues. We describe a neonate with a life-threatening airway obstruction secondary to severe micrognathia and glossoptosis. When the patient was 9 days old, bilateral distraction osteogenesis was performed to lengthen the mandible by 17 mm. The patient was subsequently extubated without further airway or swallowing difficulties. PMID- 17084234 TI - Endoscopic transnasal approach for retrieval of foreign bodies from the pterygomaxillary fossa. PMID- 17084235 TI - Bilateral peritonsillar abscesses: case report and literature review. AB - Although unilateral peritonsillar abscesses (PTA) are a common complication of acute bacterial tonsillitis, bilateral PTA are quite rare. We present the case of a 14-year-old female teenager with a 1-week history of acute tonsillitis. Physical examination revealed significant trismus, symmetrically inflamed tonsils and soft palate, and a midline uvula. A contrast-enhanced CT scan of the neck demonstrated bilateral PTA. After bilateral needle aspiration and administration of antibiotics, complete disease resolution was rapidly achieved. Patients with bilateral PTA present a diagnostic dilemma because they do not demonstrate the classic asymmetric signs and symptoms seen in the more prevalent unilateral PTA. Contrast-enhanced CT imaging can help diagnose bilateral PTA and should be considered in the presence of marked trismus but with the absence of unilateral inflammatory findings. PMID- 17084236 TI - Sonographic assessment of breast calcifications. AB - With widespread use of high-frequency sonography, breast imagers are beginning to commonly identify sonographic calcifications. The sonographic approach in assessing breast calcifications depends on whether the calcifications are initially identified sonographically or mammographically. If the calcifications are initially identified sonographically, a breast imager should first consider if the calcifications fit the characteristics of the American College of Radiology Breast Imaging Reporting and Data System Category 2--benign. If the calcifications are not benign, then one should assess the calcifications by their mammographic appearance. If the calcifications are initially identified mammographically, then the sonologist should be aware that one may not always visualize mammographic calcifications with sonography. Instead, if the radiologist seeks to localize mammographic calcifications with sonography, the imager should search for sonographic findings of invasive malignancy or ductal carcinoma in situ. These abnormalities include hypoechoic solid mass, focally dilated ducts, intraductal mass, or, rarely, isolated clustered calcifications. PMID- 17084237 TI - High-resolution computed tomographic imaging and pathologic features of pulmonary veno-occlusive disease: a review of three patients. AB - Pulmonary veno-occlusive disease (PVOD) is a rare and severe form of pulmonary hypertension that is often difficult to differentiate from primary pulmonary hypertension. Differentiating these two entities before medical treatment is critical, as therapy commonly indicated for patients with primary pulmonary hypertension can be harmful and even fatal in patients with PVOD. In the setting of known pulmonary hypertension, computed tomography findings that are highly suggestive of PVOD include extensive, patchy centrilobular ground-glass opacities, ill-defined nodular densities, and interlobular septal thickening. Definitive diagnosis requires lung biopsy, demonstrating fibrous obliteration of the pulmonary venules and small veins of the lobular septa, with secondary medial hypertrophy of the pulmonary arteries. The purpose of this article is to review reported radiographic clues to the diagnosis of PVOD, as well as to illustrate these high-resolution computed tomography findings along with pathologic correlation. PMID- 17084238 TI - Imaging of back pain in children and adolescents. AB - PURPOSE: To present the imaging findings of the wide spectrum of musculoskeletal diseases causing back pain in children and adolescents. DISCUSSION: Back pain in children is a rare condition but may denote a serious health problem; hence, full clinical history, physical examination, and appropriate laboratory studies should be obtained. In this scientific exhibit, we present the imaging findings of the variable musculoskeletal diseases that are associated with back pain in children and adolescents. These disease processes include scoliosis of various causes; spondylolysis; spondylolisthesis; traumatic injuries; disc degeneration and herniation; Scheuermann's disease; spondylodiscitis; tumors (primary, secondary, hematogenous); and miscellaneous conditions (eg, metabolic disorders, sickle cell disease, osteoporosis). CONCLUSION: A wide spectrum of diseases causing back pain in children is presented. Radiologists should be aware of the imaging findings of this rather uncommon entity to help in reaching the appropriate diagnosis. PMID- 17084239 TI - Visualization techniques in computed tomographic coronary angiography. AB - Cardiac computed tomography (CT) scans can produce enormous datasets (>3,000 images), making traditional axial image interpretation un-workable. Previously existing and new techniques including multiplanar reformats, volume rendering, maximum intensity projections, curved multiplanar reformats, and "4D" (time resolved) methods have all been used to increase diagnostic accuracy and reduce interpretation times. The methods and applications of these techniques to cardiac anatomy will be reviewed and discussed with an eye toward practical film interpretation. PMID- 17084240 TI - Cerebral varix simulating a meningioma: use of 3D magnetic resonance venography for diagnosis. AB - A rare isolated cerebral varix of a superficial convexity cortical vein was described. On conventional magnetic resonance images, the varix was initially interpreted as a meningioma but a subsequent 3D magnetic resonance venography revealed the varix and the associated venous anatomy clearly. PMID- 17084241 TI - Carpal tunnel syndrome resulting from hydroxyapatite deposition. AB - Hydroxyapatite deposition disease (HADD) is a rare cause of carpal tunnel syndrome. This is a case report of a 55-year-old male who presented with symptoms of carpal tunnel syndrome and was subsequently found to have HADD. The diagnosis of HADD is made by the identification of poorly defined cloudy calcifications on radiography of the hand and wrist. Management is usually with nonsteroidal anti inflammatory drugs, but if the pain is refractory, more invasive methods are utilized. PMID- 17084242 TI - Cardiovascular proteomics: tools to develop novel biomarkers and potential applications. AB - Proteomics is the new systems biological approach to the study of proteins and protein variation on a large scale as a result of biological processes and perturbations. The field is undergoing a dramatic transformation, owing to the completion and annotation of the human genome as well as technological advances to study proteins on a large scale. The new science of proteomics can potentially yield novel biomarkers reflecting cardiovascular disease, establish earlier detection strategies, and monitor responses to therapy. Technological advances permit the unprecedented large-scale identification of peptide sequences in a biological sample with mass spectrometry, whereas gel-based techniques provide further refinement on the status of post-translational modification. The application of high throughput protein evaluation with a subset of predefined targets, identified through proteomics, microarray profiling, and pathway analysis in animal models and human tissues, is gaining momentum in research and clinical applications. Proteomic analysis has provided important insights into ischemic heart disease, heart failure, and cardiovascular pathophysiology. The combination of proteomic biomarkers with clinical phenotypes and genetic haplotype information can lead to a more precise diagnosis and therapy on an individual basis--the fundamental premise of "personalized medicine." PMID- 17084243 TI - Impact of the degree of peri-interventional platelet inhibition after loading with clopidogrel on early clinical outcome of elective coronary stent placement. AB - OBJECTIVES: Our prospective study tested the hypothesis that the 30-day clinical outcome of elective percutaneous catheter intervention (PCI) differs between strata defined by quartiles of platelet aggregation after loading with 600 mg clopidogrel. BACKGROUND: Platelet responses after loading with clopidogrel are highly variable. The impact of this variability on the peri-interventional risk of patients undergoing PCI has not been investigated prospectively. METHODS: Our study included 802 consecutive patients undergoing elective coronary stent placement. Before PCI, patients received a loading dose of 600 mg clopidogrel followed by 75 mg daily. Primary end point was the 30-day composite of death, myocardial infarction, and target lesion revascularization (major adverse cardiac events [MACE]). Platelet aggregation was assessed immediately before PCI by optical aggregometry (5 micromol/l adenosine diphosphate). RESULTS: During 30-day follow-up, 15 patients (1.9%) incurred MACE (3 deaths, 8 myocardial infarctions, 8 target lesion revascularizations). Quartiles of platelet aggregation were <4%, 4% to 14%, 15% to 32%, and >32%. Thirty-day MACE differed significantly (p = 0.034) between quartiles of platelet aggregation. It was 0.5% in the first quartile, 0.5% in the second, 3.1% in the third, and 3.5% in the fourth. Platelet aggregation above the median carried a 6.7-fold risk (95% confidence interval 1.52 to 29.41; p = 0.003) of 30-day MACE. Multivariable logistic regression analysis, including pertinent covariables, confirmed platelet aggregation as a significant independent predictor of 30-day MACE (adjusted odds ratio per 10% increase in platelet aggregation 1.32, 95% confidence interval 1.04 to 1.61; p = 0.026). CONCLUSIONS: The level of platelet aggregation immediately before elective coronary stenting in patients pre-treated with a high loading dose of clopidogrel is correlated with early outcome after the procedure. PMID- 17084244 TI - Platelet function assessment to predict outcomes after coronary interventions: hype or hope? PMID- 17084245 TI - A model for troponin I as a quantitative predictor of in-hospital mortality. AB - OBJECTIVES: We evaluated log-transformed troponin I as a predictor of mortality in 2 independent populations. BACKGROUND: The troponin I result is typically dichotomized by a single diagnostic cutoff. Its performance as a continuous prognostic variable has not previously been well-characterized. METHODS: We studied the first troponin I sent from the emergency department (ED) as a predictor of all-cause inpatient mortality, with retrospectively gathered data. We performed our study in 2 stages, deriving our model with data from a single medical center and validating it with data from another. Subjects included every patient who had a troponin I sent from the ED during the period from November 2002 to January 2005. We assessed prognostic independence by including other potential confounders in nested logistic regression models. The troponin assay was identical at both sites (Ortho-Clinical Diagnostics, Rochester, New York). RESULTS: There were a total of 34,227 patients (12,135 derivation and 22,092 validation). Odds ratio for mortality as a function of log10-troponin was 2.08 (95% confidence interval [CI] 1.85 to 2.32) in the derivation set and 2.07 (95% CI 1.92 to 2.24) for the validation set. Troponin I remained a strong predictor after inclusion of age, electrocardiogram normality, renal insufficiency, arrival mode, chief complaint, admission diagnosis, and abnormal vital signs into bivariate and nested multivariate models. CONCLUSIONS: The presence of any detectible troponin I at ED presentation is associated with increased inpatient mortality. In 2 distinct clinical populations, the odds of death approximately doubled with any 10-fold increase in troponin result. This held true at levels below current diagnostic cutoffs. The placement and utility of dichotomous cutoffs might merit reconsideration. PMID- 17084246 TI - Chasing troponin: how low can you go if you can see the rise? PMID- 17084247 TI - Isolated elevation in troponin T after percutaneous coronary intervention is associated with higher long-term mortality. AB - OBJECTIVES: The aim of this study was to evaluate whether, in patients with normal post-procedure CK-MB, an isolated elevation in cardiac troponin T (cTnT) predicts long-term survival. BACKGROUND: Cardiac troponin T is a sensitive and specific marker of myonecrosis. There is little known about the incidence and prognostic significance of an isolated elevation of cTnT without a rise in creatine kinase (CK)-MB following PCI. METHODS: We evaluated the outcomes of 1,949 patients from the Mayo Clinic registry who had normal pre-procedure cTnT and CK-MB, required nonemergency percutaneous coronary intervention (PCI), and had normal CK-MB after the procedure. RESULTS: An elevation in cTnT (cTnT+) was observed in 383 patients (19.6%) (median 0.04 ng/ml, interquartile range 0.03 to 0.06 ng/ml). The TnT+ status was associated with adverse clinical and angiographic characteristics, and multivessel PCI. Over the median follow-up duration of 26 months, mortality (p < 0.001) and the combined rate of death and myocardial infarction (p = 0.004) were significantly higher in cTnT+ patients. Estimated 3-year survival for those with and without cTnT elevation was 86.9% and 93.2%, respectively. By multivariate analysis, an elevation in cTnT after PCI was an independent predictor of increased long-term mortality. A doubling in the post PCI cTnT was associated with a partial hazard ratio of 1.20 (95% confidence interval 1.02 to 1.40; p = 0.023). CONCLUSIONS: An isolated minor elevation in cTnT after PCI provides long-term prognostic information regarding mortality and myocardial infarction. PMID- 17084248 TI - Measuring troponin elevation after percutaneous coronary intervention: ready for prime time? PMID- 17084249 TI - Efficacy and safety of torcetrapib, a novel cholesteryl ester transfer protein inhibitor, in individuals with below-average high-density lipoprotein cholesterol levels. AB - OBJECTIVES: This study was designed to evaluate the efficacy and safety of torcetrapib, a cholesteryl ester transfer protein (CETP) inhibitor, in subjects with low high-density lipoprotein cholesterol (HDL-C) levels. BACKGROUND: Evidence suggests HDL-C is atheroprotective. A proven mechanism for increasing the level of HDL-C is the inhibition of CETP. METHODS: A total of 162 subjects with below-average HDL-C (men <44 mg/dl; women <54 mg/dl) who were not taking lipid-modifying therapy were randomized to double-blind treatment with torcetrapib 10, 30, 60, or 90 mg/day or placebo ( approximately 30 subjects per group). RESULTS: The percent change from baseline to Week 8 with torcetrapib (least-squares mean difference from placebo) was dose-dependent and ranged from 9.0% to 54.5% for HDL-C (p < or = 0.0001 for 30 mg and higher doses) and from 3.0% to -16.5% for low-density lipoprotein cholesterol (LDL-C) (p < 0.01 for 90 mg dose). Low-density lipoprotein cholesterol lowering was less in subjects with higher (>150 mg/dl) versus lower levels of baseline triglycerides; at 60 mg, the change in LDL-C was 0.1% versus -22.2% (p < 0.0001), respectively. Particle size for both HDL and LDL increased with torcetrapib. There were no dose-related increases in the frequency of adverse events. Significant blood pressure increases were noted in 2 of 140 subjects. CONCLUSIONS: Torcetrapib resulted in substantial dose-dependent elevations in HDL-C, accompanied by moderate decreases in LDL-C at the higher doses. Torcetrapib was generally well tolerated. PMID- 17084250 TI - Efficacy and safety of torcetrapib, a novel cholesteryl ester transfer protein inhibitor, in individuals with below-average high-density lipoprotein cholesterol levels on a background of atorvastatin. AB - OBJECTIVES: This study sought to evaluate the efficacy and safety of torcetrapib in patients with low high-density lipoprotein cholesterol (HDL-C) levels receiving background atorvastatin. BACKGROUND: Elevating HDL-C levels may reduce the residual cardiovascular risk that is observed in patients treated with statin therapy. Torcetrapib (a cholesteryl ester transfer protein inhibitor) increases HDL-C and decreases low-density lipoprotein cholesterol (LDL-C). METHODS: This was a multicenter, double-blind, randomized trial. Patients with below-average HDL-C (men <44 mg/dl; women <54 mg/dl) who were eligible for statin therapy according to National Cholesterol Education Program Adult Treatment Panel III guidelines or who had LDL-C >130 mg/dl at screening entered an 8-week run-in period with atorvastatin 20 mg/day before randomization (n = 174) to torcetrapib 10, 30, 60, or 90 mg/day or placebo for 8 weeks. Atorvastatin was continued during treatment with torcetrapib. RESULTS: After 8 weeks, the percent change from baseline with torcetrapib (least-squares mean difference from placebo) ranged from 8.3% to 40.2% for HDL-C (p < or = 0.0001 for 30-mg and higher doses) and from 0.6% to -18.9% for LDL-C (p < 0.01 for 60-mg and 90-mg doses). Particle size for both HDL and LDL increased with torcetrapib. The incidence of all causality and treatment-related adverse events was similar across placebo and torcetrapib treatment groups with no evidence of a dose-related response. In some treatment groups, small increases in systolic and diastolic blood pressures were noted. CONCLUSIONS: In statin-eligible patients, torcetrapib plus background atorvastatin resulted in substantial, dose-dependent increases in HDL-C, accompanied by additional decreases in LDL-C beyond those seen with atorvastatin alone. Torcetrapib plus atorvastatin was generally well tolerated. PMID- 17084251 TI - Cholesteryl ester transfer protein inhibition: the next frontier in combating coronary artery disease? PMID- 17084252 TI - Effects of high-dose atorvastatin on cerebrovascular events in patients with stable coronary disease in the TNT (treating to new targets) study. AB - OBJECTIVE: We sought to assess the effects on cerebrovascular events of treating patients with stable coronary disease with low-density lipoprotein cholesterol (LDL-C) levels substantially below 100 mg/dl. BACKGROUND: Lowering LDL-C with statins has been shown to reduce the risk of stroke in patients with stable coronary disease. In observational studies, naturally low cholesterol levels have been associated with an increased risk of hemorrhagic stroke. The cerebrovascular benefits of treating patients with stable coronary disease to LDL-C levels substantially below 100 mg/dl have not been previously investigated. METHODS: We describe an analysis of cerebrovascular events in the Treating to New Targets study, a trial where 10,001 patients with documented coronary disease were randomized to treatment with atorvastatin at 10 mg/day or 80 mg/day and followed for a median of 4.9 years. RESULTS: Mean LDL-C levels were 101 mg/dl on 10 mg atorvastatin and 77 mg/dl on 80 mg. In addition to the reduction in major cardiovascular events (hazard ratio 0.78, 95% confidence interval [CI] 0.69 to 0.89; p = 0.0002), the primary end point of the trial, patients in the 80-mg arm experienced a reduction in cerebrovascular events (hazard ratio 0.77, 95% CI 0.64 to 0.93; p = 0.007) and stroke (hazard ratio 0.75, 95% CI 0.59 to 0.96; p = 0.02). Each 1-mg/dl reduction in LDL-C with treatment was associated with a 0.6% relative risk reduction in cerebrovascular events (p = 0.002) and a 0.5% relative risk reduction in stroke (p = 0.041). The incidence of hemorrhagic stroke was similar in the 80-mg and 10-mg groups, 16 and 18 respectively, and the hemorrhagic strokes were distributed evenly across quintiles of achieved LDL-C during treatment. CONCLUSIONS: Among patients with established coronary disease, treating to an LDL-cholesterol substantially below 100 mg/dl with 80 mg/day atorvastatin reduces both stroke and cerebrovascular events by an additional 20% to 25% compared with the 10 mg/day dose. An increase in hemorrhagic stroke was not seen at low LDL-C levels. (Treating to New Targets; http://www.clinicaltrials.gov; NCT00327691). PMID- 17084253 TI - Metabolic syndrome and early-onset coronary artery disease: is the whole greater than its parts? AB - OBJECTIVES: We sought to examine the association between the metabolic syndrome (MetS) (defined both by the 2001 National Cholesterol Educational Program Adult Treatment Panel III [ATP-III] definition and the American Heart Association/National Heart, Lung and Blood Institute [AHA/NHLBI] revision incorporating the lower threshold for impaired fasting glucose [IFG]) and early onset coronary artery disease (CAD). BACKGROUND: The impact of MetS on premature CAD has not been studied extensively. Lowering the threshold to define the IFG component (from 110 to 100 mg/dl) and the value of the syndrome as a whole versus its individual components are subjects of intense debate. METHODS: We performed a case-control study with 393 early-onset CAD subjects (acute myocardial infarction, angina with > or =50% stenosis, or coronary revascularization) in men under age 46 years or women under age 56 years and 393 control subjects individually matched for gender, age, and race/ethnicity. RESULTS: By conditional logistic regression, presence of ATP-III MetS without diabetes (adjusted odds ratio [adj-OR] 4.9; 95% confidence interval [CI] 3.4 to 8.0) and with diabetes (adj-OR 8.0, 95% CI 4.39 to 14.6) was a strong independent determinant of early onset CAD. Using the AHA/NHLBI revision, these ORs became slightly stronger. However, neither definition of MetS remained significantly associated with early onset CAD in multivariate models adjusting for individual components. CONCLUSIONS: The presence of MetS imparts a high risk of early-onset clinical CAD, but the prognostic information associated with the syndrome is not greater than the sum of its parts. PMID- 17084254 TI - Gender-specific risk stratification with B-type natriuretic peptide levels in patients with acute dyspnea: insights from the B-type natriuretic peptide for acute shortness of breath evaluation study. AB - OBJECTIVES: We examined whether B-type natriuretic peptide (BNP) levels allow gender-specific risk stratification in patients with acute dyspnea. BACKGROUND: B type natriuretic peptide levels determined in patients with heart failure correlate with the severity of disease and prognosis. Gender differences in risk prediction are poorly examined. METHODS: The BASEL (B-type natriuretic peptide for Acute Shortness of Breath Evaluation) Study enrolled 190 female and 262 male patients presenting with acute dyspnea. RESULTS: At 24 months, cumulative mortality was comparable in women and men (38% vs. 35%, p = 0.66). Cox regression analyses revealed that BNP levels >500 pg/ml indicated a 5.1-fold increase in mortality for women (95% confidence interval [CI] 3.0 to 8.5, p < 0.001) versus a 1.8-fold increase in men (95% CI 1.2 to 2.6; p = 0.007). The area under the receiver-operating characteristic curve (AUC) for BNP to predict death was significantly higher in female (AUC: 0.80, 95% CI 0.73 to 0.86) than in male patients (AUC: 0.64, 95% CI 0.57 to 0.71; p = 0.001 for the comparison of AUC(women) versus AUC(men)). Women with BNP >500 pg/ml displayed a higher mortality as compared with men with BNP >500 pg/ml (68% vs. 46%, p = 0.015). Interaction analysis showed that BNP is a stronger predictor of death in women than in men (p = 0.008). CONCLUSIONS: B-type natriuretic peptide plasma levels seem to be stronger predictors of death in women than in men. PMID- 17084255 TI - Ablation of left ventricular epicardial outflow tract tachycardia from the distal great cardiac vein. AB - OBJECTIVES: The purpose of this study was to examine the feasibility and safety of ablation of idiopathic outflow tract ventricular tachycardia (OTVT) from the distal ramifications of the coronary sinus (CS). BACKGROUND: A significant minority of patients presenting with idiopathic OTVT have an epicardial focus, the standard approach to which involves ablation from within one of the aortic valve cusps (AVCs). We describe the successful ablation of idiopathic epicardial OTVT from within the CS in the distal great cardiac vein (GCV). METHODS: Ablation from the distal GCV was performed in 5 patients with idiopathic OTVT who had unfavorable mapping, in some cases unsuccessful ablation from various endocardial and epicardial sites including the AVCs, and in 1 patient via the direct epicardial approach. An electroanatomic mapping system (Carto) was used in 3 patients, and conventional mapping was performed in 2 patients, and in 3 patients cryothermal ablation was performed. RESULTS: In all patients, the first ablation lesion in the GCV successfully eliminated the arrhythmia. All patients have remained free of VT after a mean follow-up of 24 (7 to 44) months. There were no immediate or long-term complications. CONCLUSIONS: Idiopathic epicardial OTVT can be successfully ablated from the distal GCV, and should be seen as an alternative to ablation from the aortic valve cusps. PMID- 17084256 TI - In vivo 18F-fluorodeoxyglucose positron emission tomography imaging provides a noninvasive measure of carotid plaque inflammation in patients. AB - OBJECTIVES: Given the importance of inflammation in atherosclerosis, we sought to determine if atherosclerotic plaque inflammation could be measured noninvasively in humans using positron emission tomography (PET). BACKGROUND: Earlier PET studies using fluorodeoxyglucose (FDG) demonstrated increased FDG uptake in atherosclerotic plaques. Here we tested the ability of FDG-PET to measure carotid plaque inflammation in patients who subsequently underwent carotid endarterectomy (CEA). METHODS: Seventeen patients with severe carotid stenoses underwent FDG-PET imaging 3 h after FDG administration (13 to 25 mCi), after which carotid plaque FDG uptake was determined as the ratio of plaque to blood activity (target to background ratio, TBR). Less than 1 month after imaging, subjects underwent CEA, after which carotid specimens were processed to identify macrophages (staining with anti-CD68 antibodies). RESULTS: There was a significant correlation between the PET signal from the carotid plaques and the macrophage staining from the corresponding histologic sections (r = 0.70; p < 0.0001). When mean FDG uptake (mean TBR) was compared with mean inflammation (mean percentage CD68 staining) for each of the 17 patients, the correlation was even stronger (r = 0.85; p < 0.0001). Fluorodeoxyglucose uptake did not correlate with plaque area, plaque thickness, or area of smooth muscle cell staining. CONCLUSIONS: We established that FDG-PET imaging can be used to assess the severity of inflammation in carotid plaques in patients. If subsequent natural history studies link increased FDG-PET activity in carotid arteries with clinical events, this noninvasive measure could be used to identify a subset of patients with carotid atherosclerosis in need of intensified medical therapy or carotid artery intervention to prevent stroke. PMID- 17084258 TI - Non-invasive visualization of the cardiac venous system in coronary artery disease patients using 64-slice computed tomography. AB - OBJECTIVES: This study was designed to evaluate the value of 64-slice computed tomography (CT) to visualize the cardiac veins and evaluate the relation between variations in venous anatomy and history of infarction. BACKGROUND: Cardiac resynchronization therapy (CRT) is an attractive treatment for selected heart failure patients. Knowledge of venous anatomy may help in identifying candidates for successful left ventricular lead implantation. METHODS: The 64-slice CT of 100 individuals (age 61 +/- 11 years, 68% men) was studied. Subjects were divided into 3 groups: 28 control patients, 38 patients with significant coronary artery disease (CAD), and 34 patients with a history of infarction. Presence of the following coronary sinus (CS) tributaries was evaluated: posterior interventricular vein (PIV), posterior vein of the left ventricle, and left marginal vein (LMV). Vessel diameters were also measured. RESULTS: Coronary sinus and PIV were identified in all individuals. Posterior vein of the left ventricle was observed in 96% of control patients, 84% of CAD patients, and 82% of infarction patients. In patients with a history of infarction, a LMV was significantly less observed as compared with control patients and CAD patients (27% vs. 71% and 61%, respectively, p < 0.001). None of the patients with lateral infarction and only 22% of patients with anterior infarction had a LMV. Regarding quantitative data, no significant differences were observed between the groups. CONCLUSIONS: Non-invasive evaluation of cardiac veins with 64-slice CT is feasible. There is considerable variation in venous anatomy. Patients with a history of infarction were less likely to have a LMV, which may hamper optimal left ventricular lead positioning in CRT implantation. PMID- 17084257 TI - Simvastatin attenuates plaque inflammation: evaluation by fluorodeoxyglucose positron emission tomography. AB - OBJECTIVES: We investigated whether simvastatin attenuates plaque inflammation by using 18F-fluorodeoxyglucose positron emission tomography (18FDG-PET) co registered with computerized tomography. BACKGROUND: Inflammation plays a key role in progression and destabilization of atherosclerotic plaque. 18F fluorodeoxyglucose PET is a promising tool for visualizing inflammation of atherosclerotic plaque. Antiinflammatory action is one of the pleiotropic effects of statins. METHODS: Forty-three consecutive subjects, who underwent 18FDG-PET for cancer screening and had 18FDG uptakes in the thoracic aorta and/or the carotid arteries, were randomized to either statin group receiving simvastatin (n = 21) or diet group receiving dietary management only (n = 22). The maximum standardized uptake values (SUVs) were measured in individual plaques, and were averaged for analysis of the subjectwise results. The responses were assessed after 3-month treatments. RESULTS: Positron emission tomography revealed 117 and 123 18FDG-positive plaques in the statin and diet groups, respectively. Simvastatin, but not diet alone, attenuated plaque (18)FDG uptakes and decreased the SUVs (p < 0.01). Simvastatin reduced low-density lipoprotein cholesterol (LDL C) by 30% (p < 0.01) and increased high-density lipoprotein cholesterol (HDL-C) by 15% (p < 0.01), whereas LDL-C and HDL-C levels were not changed in the diet group. In the statin group, the decrease in the SUV was well correlated with the HDL-C elevation (p < 0.01) but not with the LDL-C reduction. CONCLUSIONS: 18F fluorodeoxyglucose PET visualized plaque inflammation and simvastatin attenuated it. The LDL-C-independent effects of simvastatin may participate in the beneficial effect. 18F-fluorodeoxyglucose PET has a potential for visually monitoring plaque inflammation and the therapeutic effectiveness of statins. PMID- 17084259 TI - Magnetically targeted endothelial cell localization in stented vessels. AB - OBJECTIVES: A novel method to magnetically localize endothelial cells at the site of a stented vessel wall was developed. The application of this strategy in a large animal model is described. BACKGROUND: Local delivery of blood-derived endothelial cells has been shown to facilitate vascular healing in animal models. Therapeutic utilization has been limited by an inability to retain cells in the presence of blood flow. We hypothesized that a magnetized stent would facilitate local retention of superparamagnetically labeled cells. METHODS: Cultured porcine endothelial cells were labeled with endocytosed superparamagnetic iron oxide microspheres. A 500:1 microsphere-to-cell ratio was selected for in vivo experiments based on bromo-deoxyuridine incorporation and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling assays. Stents were magnetized and implanted in porcine coronary and femoral arteries using standard interventional equipment. Labeled endothelial cells were delivered locally during transient occlusion of blood flow. RESULTS: The delivered cells were found attached to the stent struts and were also distributed within the adjacent denuded vessel wall at 24 h. CONCLUSIONS: Magnetic forces can be used to rapidly place endothelial cells at the site of a magnetized intravascular stent. The delivered cells are retained in the presence of blood flow and also spread to the adjacent injured vessel wall. Potential applications include delivering a cell based therapeutic effect to the local vessel wall as well as downstream tissue. PMID- 17084260 TI - Non-invasive assessment of endothelial function: which technique? AB - OBJECTIVES: The purpose of this study was to compare 3 non-invasive techniques for assessment of endothelial function in adults and children and evaluate their utility in acute inflammation. BACKGROUND: Endothelial dysfunction is a key early event in pre-clinical atherosclerosis. Flow-mediated dilation (FMD), although the established technique, is expensive and technically demanding. Measurements of vascular responses to inhaled salbutamol by pulse wave analysis (PWA) or pulse contour analysis (PCA) are potential alternatives. METHODS: Sixteen adults (mean age 28 years, range 18 to 39) and 16 children (mean age 13 years, range 7 to 17) underwent concurrent vascular function testing on 2 occasions with ultrasound, PWA, and PCA. Eighteen men were also studied before and after typhoid vaccination. RESULTS: Reproducibility of FMD was high in adults and children (coefficient of variation [CV] = 7.1 and 6.3, respectively). Salbutamol responses were more variable with PWA (adults CV = 11.5, children CV = 17.1) and PCA particularly in children (adults CV = 18.2, children CV = 36.3). Flow-mediated dilation (p < 0.001) and PWA with salbutamol (p = 0.03) responses fell after typhoid vaccination, and PCA (p = 0.7) was unchanged. CONCLUSIONS: Vascular dysfunction during acute inflammation can be measured by FMD and by PWA with salbutamol. Flow-mediated dilation is less variable than PWA. Variability of PCA makes this technique currently unsuited to serial measures of endothelial function in children. Flow-mediated dilation remains the most reproducible method. PMID- 17084261 TI - Assessment of renal artery stenosis severity by pressure gradient measurements. AB - OBJECTIVES: The purpose of this study was to define "significant" renal artery stenosis (i.e., a stenosis able to induce arterial hypertension). BACKGROUND: The degree of renal artery stenosis that justifies an attempt at revascularization is unknown. METHODS: In 15 patients, transstenotic pressure measurements were obtained before and after unilateral stenting. After stenting, graded stenoses were created in the stented segment by progressive inflation of a balloon catheter. Stenosis severity was expressed as the ratio of distal pressure (P(d)) corrected for aortic pressure (P(a)). Balloon inflation pressure was adjusted to create 6 degrees of stenosis (P(d)/P(a) from 1.0 to 0.5, each step during 10 min). Plasma renin concentration was measured at the end of each step in the aorta and in both renal veins. RESULTS: For a P(d)/P(a) ratio >0.90, no significant change in plasma renin concentration was observed. However, when P(d)/P(a) became <0.90, a significant increase in renin was observed in the renal vein of the stenotic kidney, finally reaching a maximal increase of 346 +/- 145% for P(d)/P(a) of 0.50 (p = 0.006). These values returned to baseline when the stenosis was relieved. In addition, plasma renin concentration increased significantly in the vein from the non-stenotic kidney (p = 0.02). CONCLUSIONS: In renal artery stenoses, a P(d)/P(a) ratio of 0.90 can be considered a threshold value below which the stenosis is likely responsible for an up-regulation of renin production and, thus, for renovascular hypertension. These findings might contribute to better patient selection for renal angioplasty. PMID- 17084262 TI - Physiologic assessment of renal artery stenosis: will history repeat itself? PMID- 17084263 TI - Adverse effects of dopamine on systemic hemodynamic status and oxygen transport in neonates after the Norwood procedure. AB - OBJECTIVES: The purpose of this study was to evaluate the effects of dopamine on hemodynamic status and oxygen transport in neonates after the Norwood procedure. BACKGROUND: Dopamine is widely used to augment cardiac performance and increase oxygen delivery (DO2) in patients after cardiopulmonary bypass (CPB). This might be at the expense of increased myocardial and systemic oxygen consumption (VO2), thus offsetting the improved DO2. This balance is particularly fragile in critically ill neonates. METHODS: Systemic oxygen consumption was continuously measured with respiratory mass spectrometry in 13 sedated, paralyzed, and mechanically ventilated neonates for 72 h after the Norwood procedure. Arterial, superior vena caval, and pulmonary venous blood gases were measured to calculate pulmonary blood flow (Q(p)) and systemic blood flow (Q(s)), DO2, and oxygen extraction ratio (ERO2). Rate-pressure product was calculated. Dopamine at a dose of 5 microg/kg/min was routinely administered at cessation of CPB and terminated within the first 48 h. Hemodynamic and oxygen transport measures were obtained before and at 100 min after the termination of dopamine. RESULTS: Terminating dopamine was not associated with significant changes in arterial pressure, Q(p), Q(s), or DO2 but was associated with a significant decrease in heart rate (p = 0.003), rate-pressure product (p = 0.03), and VO2 (-20 +/- 11%, p < 0.0001), resulting in a significant decrease in ERO2 (p = 0.01). CONCLUSIONS: Dopamine induces a significant increase in VO2 in neonates after the Norwood procedure, and termination is associated with an improved balance of VO2-DO2. These data further emphasize the importance of understanding changes in VO2 as well as DO2 in infants after cardiac surgery. PMID- 17084264 TI - Improvement of early vascular changes and cardiovascular risk factors in obese children after a six-month exercise program. AB - OBJECTIVES: The present study aimed to assess the effect of a 6-month exercise program in obese children on flow-mediated vasodilation (FMD) carotid intima media thickness (IMT) and cardiovascular risk factors (RF). BACKGROUND: Childhood obesity contributes to adult obesity and subsequent cardiovascular disease. Physical inactivity is a major RF for obesity, endothelial dysfunction, and elevated carotid IMT, culminating in early atherosclerotic disease. METHODS: Sixty-seven obese subjects (age 14.7 +/- 2.2 years) were randomly assigned to 6 months' exercise or non-exercise protocol. We examined the influence of exercises (1 h, 3 times/week) on FMD, IMT, and cardiovascular risk profile. RESULTS: Compared with lean control subjects, obese children demonstrated at baseline significantly impaired FMD (4.09 +/- 1.76% vs. 10.65 +/- 1.95%, p < 0.001), increased IMT (0.48 +/- 0.08 mm vs. 0.37 +/- 0.05 mm, p < 0.001), and a number of obesity-related cardiovascular RF. Significant improvements were observed in the exercise group for IMT (0.44 +/- 0.08 mm, p = 0.012, -6.3%) and FMD (7.71 +/- 2.53%, p < 0.001, +127%). This improvement correlated with reduced RF, such as body mass index standard deviation scores, body fat mass, waist/hip ratio, ambulatory systolic blood pressure, fasting insulin, triglycerides, low-density lipoprotein/high-density lipoprotein ratio, and low-degree inflammation (C reactive protein, fibrinogen). CONCLUSIONS: The present study documented increased IMT, impaired endothelial function, and various elevated cardiovascular RF in young obese subjects. Regular exercise over 6 months restores endothelial function and improves carotid IMT associated with an improved cardiovascular risk profile in obese children. PMID- 17084265 TI - A novel inhibitory effect of naloxone on macrophage activation and atherosclerosis formation in mice. AB - OBJECTIVES: We investigated whether naloxone could reduce macrophage activation and influence atherosclerotic lesion formation in mice. BACKGROUND: Macrophages play an important role in the inflammatory process in atherosclerosis. Naloxone could inhibit activation of microglia, the resident macrophage in the nervous system. METHODS: The anti-inflammatory effect of naloxone was evaluated by stimulating the macrophage cell culture and FVB mice with lipopolysaccharide or oxidized low-density lipoprotein with and without naloxone pretreatment. Apolipoprotein-E (apoE)-deficient mice received naloxone injection for 10 weeks, and the severity of aortic atherosclerosis was measured. The left common carotid arteries of C57BL/6 mice were ligated near the carotid bifurcation. The mice then received naloxone injection for 4 weeks after ligation, and the severity of neointima formation was evaluated. RESULTS: Naloxone pretreatment significantly suppressed the production of tumor necrosis factor-alpha (TNF-alpha), interleukin 6, monocyte chemoattractant protein-1, and superoxide in macrophages after stimulation. In FVB mice, naloxone reduced the TNF-alpha level in circulation, inflammatory cell infiltration in lungs, and superoxide production in aorta. Naloxone injection significantly decreased the severity of aortic atherosclerosis in the apoE-deficient mice and carotid neointima formation in the C57BL/6 mice after ligation. CONCLUSIONS: Naloxone, with its novel anti-inflammatory effect, significantly reduces atherosclerosis and neointima formation in mice. PMID- 17084266 TI - Meeting highlights of the 17th annual scientific sessions of the American Society of Echocardiography. Baltimore, Maryland, June 3-7, 2006. PMID- 17084267 TI - The year in epidemiology, health services research, and outcomes research. PMID- 17084268 TI - Diagnostic performance of multislice spiral computed tomography of coronary arteries as compared with conventional invasive coronary angiography: a meta analysis. AB - OBJECTIVES: This study was designed to define the current role of multislice spiral computed tomography (MSCT) for the diagnosis of coronary artery disease (CAD) using a meta-analytic process. BACKGROUND: Multislice spiral computed tomography has recently been proposed as an alternative to conventional coronary angiography (CA) for the diagnosis of CAD. METHODS: Using Medline, we identified 29 studies (2,024 patients) evaluating CAD by means of both MSCT (> or =16 slices) and conventional CA before July 2006. After data extraction the analysis was performed according to a random-effects model. RESULTS: The per-segment analysis pooled the results from 27 studies corresponding to a cumulative number of 22,798 segments. Among unassessable segments, 4.2% were excluded from the analysis and 6.4% were classified at the discretion of the investigators, underscoring the shortcomings of MSCT. With this major limitation, the per segment sensitivity and specificity were 81% (95% confidence interval [CI] 72% to 89%) and 93% (95% CI 90% to 97%), respectively, with positive and negative likelihood ratios of 21.5 (95% CI 13.1 to 35.5) and 0.11 (95% CI 0.06 to 0.21), respectively, and positive and negative predictive values of 67.8% (95% CI 57.6% to 78.0%) and 96.5% (95% CI 94.7% to 98.3%), respectively. As expected, the per patient analysis has shown an increased sensitivity of 96% (95% CI 94% to 98%) but a decreased specificity of 74% (95% CI 65% to 84%). CONCLUSIONS: Multislice spiral computed tomography has shortcomings difficult to overcome in daily practice and, at the more clinically relevant per-patient analysis, continues to have moderate specificity in patients with high prevalence of CAD. Studies evaluating the diagnostic performance of the newest generation of MSCT, including patients with low to moderate CAD prevalence, will be critical in establishing the clinical role of this emerging technology as an alternative to CA. PMID- 17084269 TI - President's page: GAP-D2B: an alliance for quality. PMID- 17084270 TI - Spontaneous late closure of patent foramen ovale. PMID- 17084271 TI - Patent foramen ovale and the risk of cryptogenic stroke. PMID- 17084273 TI - Coronary plaque burden and cardiovascular risk factors: single-point versus serial assessment. PMID- 17084276 TI - Most asymptomatic diabetic patients will not benefit from coronary revascularization. PMID- 17084280 TI - Reversal of calcium cycling defects in advanced heart failure toward molecular therapy. AB - Heart failure is a growing major cause of human morbidity and mortality worldwide. A wave of new insights from diverse laboratories has begun to uncover new therapeutic strategies that affect the molecular pathways within cardiomyocytes that drive heart failure progression. Using an integrative approach that employs insights from genetic-based studies in mouse and humans and in vivo somatic gene transfer studies, we have uncovered a new link between stress signals mediated by mechanical stretch and defects in sarcoplasmic reticulum (SR) calcium cycling. An intrinsic mechanical stress sensing system is embedded in the Z disc of cardiomyocytes, and defects in stretch responses can lead to heart failure progression and associated increases in wall stress. Reversal of the chronic increases in wall stress by promoting SR calcium cycling can prevent and partially reverse heart failure progression in multiple genetic and acquired model systems of heart failure in both small and large animals. We propose that reversal of advanced heart failure is possible by targeting the defects in SR calcium cycling, which may be a final common pathway for the progression of many forms of heart failure. PMID- 17084284 TI - Protein kinase C beta/early growth response-1 pathway: a key player in ischemia, atherosclerosis, and restenosis. AB - Atherosclerosis, restenosis, and the consequences of ischemia are the major causes of morbidity and mortality worldwide. Elucidation of key contributing pathways in animal models of ischemia-reperfusion injury, atherosclerosis, and restenosis consequent to vascular injury may lead to great interest in determining if blocking these pathways could prevent vascular disease in human subjects. This review details the evidence that the protein kinase C (PKC) beta/early growth response-1 axis plays a central role in the response to both acute and chronic vascular stresses in animal models and also indicates the clinical implications of a specific inhibitor of PKCbeta, ruboxistaurin (LY333531). PMID- 17084288 TI - Mechanical and handling properties of braided polyblend polyethylene sutures in comparison to braided polyester and monofilament polydioxanone sutures. AB - PURPOSE: This study was designed to comprehensively compare the mechanical properties of 4 types of braided polyblend sutures with widely used braided polyester and monofilament polydioxanone sutures. METHODS: Polyblend polyethylene sutures (FiberWire [Arthrex, Naples, FL], Herculine [Linvatec, Largo, FL], Orthocord [DePuy Mitek, Raynham, MA], and Ultrabraid [Smith & Nephew Endoscopy, Andover, MA]), a braided polyester suture (Ethibond; Ethicon, Somerville, NJ), and an absorbable monofilament polydioxanone suture (PDS II; Ethicon), all USP No. 2, were mechanically tested. Fraying resistance was tested on eyelets of metallic and absorbable suture anchors. Cartilage abrasion caused by an intra articularly placed suture knot was simulated by fraying on distal porcine femora. RESULTS: All polyblend sutures were stronger than Ethibond or PDS II sutures by at least a factor of 2, with or without a knot. When knotted, Herculine (261 +/- 44 N) was strongest, followed by Ultrabraid (244 +/- 3 N). FiberWire was most resistant against fraying on metallic anchors. Orthocord was by far least abrasive with absorbable anchors. Resistance to fraying was 100- to 500-fold (absorbable anchors) and 6- to 30-fold (metallic anchor) better for all polyblend sutures than for Ethibond. All braided sutures caused a similar amount of abrasion of joint cartilage, but they caused significantly more abrasion (>20 fold) than the monofilament degradable suture. CONCLUSIONS: The ultimate strength of polyblend suture material was 2- to 2.5-fold greater than that of polyester or polydioxanone sutures, but the resistance to fraying was up to 500-fold greater than that of polyester or polydioxanone sutures. With regard to strength, this makes polyblend sutures particularly advantageous for use with metallic edges of anchors or prostheses or with absorbable anchor eyelets. CLINICAL RELEVANCE: With a high resistance to fraying against metallic edges or a decrease in cutting of absorbable suture eyelets being up to 500-fold greater than with polyester or polydioxanone sutures, the new polyblend sutures appear to fill a void in the armamentarium of the surgeon, provided that at least 2 throws more than with conventional sutures are used for knot tying. PMID- 17084289 TI - Triple-loaded single-anchor stitch configurations: an analysis of cyclically loaded suture-tendon interface security. AB - PURPOSE: This study evaluated the strength and suture-tendon interface security of different suture configurations from triple-suture-loaded anchors. METHODS: A juvenile bovine infraspinatus tendon was detached and repaired by use of 4 different suture combinations from 2 suture anchors: 3 simple sutures in each anchor (ThreeVo anchor; Linvatec, Largo, FL); 2 peripheral simple stitches and 1 central horizontal mattress suture passed deeper into the tendon, creating a larger footprint (bigfoot-print anchor); 2 peripheral simple stitches with 1 central horizontal mattress stitch passed through the same holes as the simple sutures (stitch-of-Burns); and 2 simple stitches (TwoVo anchor; Linvatec). The constructs were cyclically loaded between 10 N and 180 N for 3,500 cycles and then destructively tested. The number of cycles required to create a 5-mm gap and a 10-mm gap and the ultimate load to failure and failure mode were recorded. RESULTS: The ThreeVo anchor was strongest and most resistant to cyclic loading (P < .01). The TwoVo anchor was least resistant to cyclic loading. The stitch-of Burns anchor was more resistant to cyclic loading than both the bigfoot-print anchor and the TwoVo anchor (P < .03). The ThreeVo, stitch-of-Burns, and TwoVo anchors were stronger than the bigfoot-print anchor (P < .05). CONCLUSIONS: Three simple sutures in an anchor hold better than two simple sutures. Three simple sutures provide superior suture-tendon security than combinations of one mattress and two simple stitches subjected to cyclic loading. A central mattress stitch placed more medially than two peripheral simple stitches (bigfoot-print anchor) configured to enlarge the tendon-suture footprint was not as resistant to cyclic loading or destructive testing as three simple stitches (ThreeVo anchor). CLINICAL RELEVANCE: Placing a central mattress stitch more medially than 2 peripheral simple stitches to enlarge the tendon-suture footprint was not as resistant to cyclic loading or destructive testing as 3 simple stitches. PMID- 17084290 TI - An ex vivo thermal chondroplasty model: the association of a char-like layer and underlying cell death. AB - PURPOSE: The purpose of this study was to evaluate the relation between the char like layer observed during radiofrequency energy (RFE) treatment of cartilage and the depth of underlying cell death. METHODS: Healthy adult bovine patellae were treated with a monopolar RFE probe ex vivo at generator settings of 20, 30, 40, 50, 60, 80, and 110 in cut mode. The presence or absence of a char-like layer and visual electrical discharge was noted. Treated tissue was incubated with cell viability stain, and the depth of cell death and matrix debridement was measured from confocal laser microscopy images. RESULTS: At generator settings of 60 and above, a char-like layer, electrical discharge, and matrix debridement were consistently observed, and the depth of cell death was significantly less (P < .05) than when these features were not observed (< or =30). Paradoxically, the least depth of cell death did not occur at the lowest generator setting in cut mode. It occurred at a generator setting of 60. An increase in impedance of the system and a decrease in current were also associated with reduced cell death. CONCLUSIONS: In this controlled ex vivo study formation of a char-like layer, visual electrical discharge, increased impedance, and reduced current were associated with less depth of cell death when cartilage was treated with monopolar RFE. CLINICAL RELEVANCE: This study suggests that a char-like layer and electrical discharge during RFE treatment of cartilage may be advantageous because, potentially, these features are associated with less depth of cell death (safety) and greater matrix debridement (efficacy). PMID- 17084291 TI - Quantifying the extent of a type II SLAP lesion required to cause peel-back of the glenoid labrum--a cadaveric study. AB - PURPOSE: To quantify the extent of labral disruption required to cause it to peel back when the peel-back test is performed. METHODS: Ten cadaveric shoulders were prepared by removal of the deltoid and rotator cuff muscles. The glenohumeral joint was concentrically reduced and brought into 90 degrees abduction and maximal external rotation. The peel-back of the labrum was graded 0, 1, or 2. The labrum was sequentially detached from the glenoid in the following order: biceps anchor only, 1 o'clock, 2 o'clock, 11 o'clock, and 3 o'clock positions. After each labral cut, the peel-back test was performed. Labral repair was performed with a single suture anchor placed at the 12:30 o'clock position; labral peel back was reassessed. RESULTS: A progressive increase was noted in peel-back grade with sequential cutting of the labrum posteriorly. However, disruption of the anchor alone did not lead to a positive peel-back sign. Disruption to the 2 o'clock position resulted in a positive peel-back sign overall in 9 of 10 shoulders (5 were grade 1, and 4 were grade 2). No increase was seen in peel-back grade with anterior extension of the labral detachment. Labral repair with a single anchor placed at the 12:30 o'clock position eliminated labral peel-back in 100% of shoulders. CONCLUSIONS: Detachment of the biceps anchor alone does not cause peel-back. The labrum must be disrupted to at least the 2 o'clock position before overt (grade 2) peel-back is observed. A single suture anchor placed at 12:30 o'clock eliminated peel-back of the labrum. CLINICAL RELEVANCE: Validation of the peel-back test as an important diagnostic tool during shoulder arthroscopy. PMID- 17084292 TI - The effect of double-row fixation on initial repair strength in rotator cuff repair: a biomechanical study. AB - PURPOSE: The purpose of this study was to compare the initial mechanical strength of 3 rotator cuff repair techniques. METHODS: A total of 30 fresh-frozen cadaveric shoulders were prepared, and full-thickness supraspinatus tears were created. Specimens were randomized and placed into 3 groups: (1) transosseous suture technique (group I: TOS, n = 10, 6F/4M), (2) single-row suture anchor fixation (group II: SRSA, n = 10, 6F/4M), and (3) double-row suture anchor fixation (group III: DRSA, n = 10, 6F/4M). Each specimen underwent cyclic load testing from 5 N to 180 N at a rate of 33 mm/sec. The test was stopped when complete failure (repair site gap of 10 mm) or a total of 5,000 cycles was attained. RESULTS: Group I (TOS) failed at an average of 75.3 +/- 22.49 cycles, and group II (SRSA) at an average of 798.3 +/- 73.28 cycles; group III (DRSA) had no failures because all samples were stopped when 5,000 cycles had been completed. Fixation strength of the DRSA technique proved to be significantly greater than that of SRSA (P < .001), and both suture anchor groups were significantly stronger than the TOS group (P < .001). CONCLUSIONS: Suture anchor repairs were significantly stronger than transosseous repairs. Furthermore, double-row suture anchor fixation was significantly stronger than was single-row repair. Therefore, double-row fixation may be superior to other techniques in that it provides a substantially stronger repair that could lead to improved biologic healing. CLINICAL RELEVANCE: A high incidence of incomplete healing occurs in rotator cuff repair. Use of double-row fixation may help the clinician to address some deficiencies in current methods by increasing the strength of the repair, potentially leading to improved healing rates. PMID- 17084293 TI - A clinical composite score accurately detects meniscal pathology. AB - PURPOSE: Five common tests were used to diagnose meniscal tears when used as a composite score. We evaluated how effectively the composite examination, when performed in the presence of an anterior cruciate ligament (ACL) injury or degenerative joint disease (DJD), determined the presence of meniscal tears. METHODS: Data were collected prospectively on all patients at our clinic with a primary knee complaint. Independent variables included the presence or absence of the following: (1) a history of "catching" or "locking" as reported by the patient, (2) pain with forced hyperextension, (3) pain with maximum flexion, (4) pain or an audible click with McMurray's maneuver, and (5) joint line tenderness to palpation. Comprehensive patient demographic data were collected including ligamentous examinations and other intra-articular pathologies found at arthroscopy. Composite examination findings were correlated with the presence or absence of meniscal pathology. RESULTS: We evaluated 635 knees in 576 patients for historical and physical findings. Of the knees, 209 underwent arthroscopic surgery and 426 did not. Chi(2) Analysis showed a significant relation between the number of positive diagnostic tests and the presence of meniscal tears (P = .001). Five positive findings on composite examination yielded a positive predictive value of 92.3%. Positive predictive values remained greater than 75% with composite scores of at least 3 in the absence of ACL and DJD pathologies. The presence of an ACL injury decreased the positive predictive value of 5 composite findings to 67%, whereas the presence of DJD increased predictability to 100%. CONCLUSIONS: When all 5 symptoms and signs were positive, there was a 92.3% positive predictive value of finding a meniscal tear. Although positive predictive values decreased with a concomitant ACL injury and increased with DJD, there was a higher rate of false-positive findings (ACL) and false-negative findings (DJD). LEVEL OF EVIDENCE: Level II, development of diagnostic criteria with consecutive patients and gold standard. PMID- 17084294 TI - Is microfracture of chondral defects in the knee associated with different results in patients aged 40 years or younger? AB - PURPOSE: Age-dependent studies about the clinical result after microfracture of cartilage lesions in the knee are still missing. This prospective study was performed to discover age-dependent differences in the results after microfracture over a period of 36 months. METHODS: Between 1999 and 2002, 85 patients (mean age, 39 years) with full-thickness chondral lesions underwent the microfracture procedure and were evaluated preoperatively and at 6, 18, and 36 months after surgery. Depending on the patients' age (< or =40 years or >40 years) and the localization of the defects (femoral condyles, tibia, and patellofemoral joint), the patients were assigned to 6 different groups. Exclusion criteria were meniscal pathologic conditions, tibiofemoral malalignment, and ligament instabilities. Baseline clinical scores were compared with follow-up data by use of paired Wilcoxon tests for the modified Cincinnati knee score and the International Cartilage Repair Society (ICRS) score. RESULTS: The scores improved in all groups over the whole study period (P < .05). Patients aged 40 years or younger had significantly better results (P < .01) for both scores compared with older patients. Between 18 and 36 months after microfracture, the ICRS score deteriorated significantly (P < .05) in patients aged over 40 years whereas younger patients with defects on the femoral condyles and on the tibia showed neither a significant improvement nor a significant deterioration in the ICRS score (P > .1). Magnetic resonance imaging 36 months after surgery revealed better defect filling and a better overall score in younger patients (P < .05). The Spearman coefficient of correlation between clinical and magnetic resonance imaging scores was 0.84. CONCLUSIONS: The clinical results after microfracture of full-thickness cartilage lesions in the knee are age-dependent. Deterioration begins 18 months after surgery and is significantly pronounced in patients aged older than 40 years. The best prognostic factor was found to be a patient age of 40 or younger with defects on the femoral condyles. LEVEL OF EVIDENCE: Level IV, prognostic case series. PMID- 17084295 TI - Ulnar collateral ligament reconstruction in elite throwing athletes. AB - PURPOSE: Ulnar collateral ligament (UCL) injuries may result in disabling valgus instability in throwing athletes. We evaluated the docking technique for UCL reconstruction and describe a modification to the technique. METHODS: UCL surgery was indicated in 20 high-level baseball players (13 professional and 7 collegiate) based on medial elbow pain preventing effective throwing, clinically apparent medial elbow laxity, and magnetic resonance arthrogram consistent with UCL injury. The mean age was 21.7 years (range, 17.9 to 25.3 years). One patient had previous UCL reconstruction. One had previous arthroscopic elbow debridement. The mean time between injury and treatment was 73 days. Reconstruction was performed via a muscle-splitting approach and the docking technique with palmaris or gracilis graft. For the initial 12 patients, a 2-strand construct was used; however, during the study period, we developed and began using a 3-strand construct with a double anterior bundle and a single posterior bundle, which was used in the next 8 patients. The ulnar nerve was not routinely transposed unless there were preoperative ulnar nerve symptoms (1 patient). Two patients had osteophyte debridement. One had removal of a loose body. RESULTS: Patients were followed up for a mean of 41.9 months (range, 6.4 to 67.1 months). One player was lost to follow-up and could not be identified on a professional roster. Of the remaining 19 patients, 18 returned to their previous level of participation or higher. Two were occasional pitchers who did not wish to return to pitching but continued to play other positions. They were clinically and functionally asymptomatic. The mean time to return to play was 13.1 months (range, 6.3 to 21.3 months). By use of the Timmerman-Andrews 100-point subjective scoring system, the mean preoperative score was 77.0 (range, 65 to 80) and the mean postoperative score was 98.2 (range, 85 to 100). By use of the Conway-Jobe scoring system, the outcome was rated as excellent in 17 patients and good in 2. One patient underwent subsequent ulnar nerve transposition and returned to the previous level of professional play. CONCLUSIONS: UCL reconstruction with the docking technique is a reproducible and safe operation that can reliably return athletes to a high level of participation with limited adverse effects. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 17084296 TI - Arthroscopic medial retinacular repair after patellar dislocation with and without underlying trochlear dysplasia: a preliminary report. AB - PURPOSE: This study was undertaken to evaluate the influence of underlying trochlear dysplasia (TD) on clinical outcomes of arthroscopic medial retinacular repair. METHODS: Between January 2000 and October 2004, a total of 91 patients underwent arthroscopic medial retinacular repair. Inclusion criteria for this study included an arthroscopic medial retinacular repair, a follow-up time of 12 months, and trochlear grading based on axial computed tomography (CT) scans (n = 48). TD, if present, was graded as types A through D, and patients were separated into group I (no or low-grade type A TD) and group II (types B through D TD). Redislocation was recorded, and Tegner, Lysholm, and International Knee Documentation Committee (IKDC) scores were determined. RESULTS: No or grade A TD was detected in 26 knees (group I), and TD of grade B or C was found in 22 knees (group II). Type D TD was not observed. At a follow-up time of 12 months, 4 redislocations had been noted, all belonging to group II. Neither the Tegner nor the Lysholm score reached preinjury levels, but in a comparison with preoperative status, we could find a significant increase in all scores in both groups. Group I reached a significantly better postoperative outcome than was attained by group II. CONCLUSIONS: Arthroscopic repair of the medial retinaculum is an effective technique by which patellofemoral instability can be addressed when normal or nearly normal trochlear geometry is present. In patients with underlying TD, patellofemoral stability cannot be completely restored, and clinical results are less successful. Precise preoperative radiologic determination of trochlear geometry may help the clinician to predict short-term outcomes in patients with patellofemoral instability. LEVEL OF EVIDENCE: Level III, retrospective comparative therapeutic study. PMID- 17084297 TI - Intra-articular injection of hyaluronic acid is not effective for the treatment of post-traumatic osteoarthritis of the elbow. AB - PURPOSE: In this study we evaluated the effect and safety of intra-articular injection of hyaluronic acid in 19 consecutive elbows with post-traumatic osteoarthritis. METHODS: In 18 patients (10 male and 8 female patients; mean age, 45.6 years [SD, 15.0 years]), 3 injections of sodium hyaluronate were given within 4 weeks at regular intervals. Evaluation took place just before the first injection, as well as after 3 and 6 months, and consisted of the Elbow Function Assessment Score, the Functional Rating Index of Broberg and Morrey, and the Modified Andrews Elbow Scoring System. Pain was also assessed by means of visual analog scales. RESULTS: Viscosupplementation resulted in slight, short-term pain relief and a very limited decrease in activity impairment at evaluation after 3 months. After 6 months, no beneficial effects were noticed in any of the 19 injected elbows. Other parameters were not influenced by treatment with viscosupplementation at any time. Systemic or local adverse effects did not occur. CONCLUSIONS: Because the use of viscosupplementation for the treatment of post-traumatic osteoarthritis of the elbow provides only slight, short-term pain relief and a very limited decrease in activity impairment and the other parameters were not modified, we believe that viscosupplementation is not suitable for this indication. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 17084298 TI - Graft laceration and pullout strength of soft-tissue anterior cruciate ligament reconstruction: in vitro study comparing titanium, poly-d,l-lactide, and poly-d,l lactide-tricalcium phosphate screws. AB - PURPOSE: The aim of part 1 of this study was to compare the structural properties of tibia-graft-interference screw constructs of soft-tissue graft fixation via the anterior cruciate ligament (ACL) by use of different interference screws. In part 2 the influence of graft laceration as a result of 1 single interference screw insertion on the mechanical properties of the graft was evaluated. METHODS: In part 1 of the study, in a bovine knee model, soft-tissue graft fixation with titanium, biodegradable poly-D,L-lactide (PLDLA), and PLDLA-tricalcium phosphate (TCP) interference screws was performed, and the structural properties were determined by use of a materials testing machine at a rate of 12.5 mm/s. In part 2 the bone was cut after 1 single insertion of titanium, PLDLA, or PLDLA-TCP interference screws, and the mechanical properties of the graft were determined. RESULTS: Soft-tissue graft fixation with a biodegradable PLDLA interference screw showed a statistically significantly higher maximum load when compared with PLDLA TCP and titanium interference screws. PLDLA-TCP screw fixation revealed a significantly higher ultimate load than titanium screw fixation. The mechanical properties of the grafts after 1 single insertion of PLDLA-TCP interference screws showed a statistically significantly higher stiffness, yield load, and maximum load when compared with the PLDLA and titanium interference screw groups. CONCLUSIONS: The results of this study suggest that the use of a titanium screw to fix a soft-tissue graft in cruciate ligament reconstruction surgery may cause damage to the graft. Soft-tissue ACL reconstruction fixed with a PLDLA interference screw resulted in a significantly higher pullout strength than ACL reconstructions fixed with a PLDLA-TCP or titanium interference screw. CLINICAL RELEVANCE: In contrast to fixation with a biodegradable interference screw, fixation with a titanium interference screw may damage the integrity of a soft tissue graft in cruciate ligament reconstruction and should therefore be avoided. PMID- 17084299 TI - Intraoperative incidents and complications in primary arthroscopic anterior cruciate ligament reconstruction. AB - PURPOSE: To analyze intraoperative incidents and complications in primary arthroscopic anterior cruciate ligament reconstruction (ACLR) with bone-patellar tendon-bone (BPTB) and quadruple hamstring semitendinosus and gracilis (STG) grafts. METHODS: From January 1999 to June 2004, we performed 330 BPTB and 96 STG procedures. Intraoperative incidents and complications were recorded for each case. RESULTS: Our overall intraoperative incident and complication rate was 9.6% and was higher in the STG group (13.5%) than in the BPTB group (8.7%). In the BPTB group, combined incidents and complications according to stage of the procedure consisted of graft harvesting, 4 cases (1.21%); tunnel placement, 9 cases (2.7%); and graft fixation, 15 cases (4.5%). In the STG group, combined incidents and complications included graft harvesting, 8 cases (8.3%); tunnel placement, 1 case (1%); graft fixation, 3 cases (3.1%); and others, 1 case (1%). In 4 patients (0.9%), we had to change the technique from BPTB to STG and vice versa. In 2 STG cases, conversion of the technique was required because of graft related problems; in the third case, it was necessary because of fixation. One BPTB was converted to STG because the graft was completely transected as a result of improper screw fixation technique. CONCLUSIONS: None of the encountered incidents and complications had an adverse effect on final stability or on range of motion in operated knees. All incidents and complications were due to technical errors that occurred during graft harvesting, tunnel placement, or graft fixation. It is known that technical errors are preventable if careful surgical technique is followed. The surgeon must be well versed in various reconstruction techniques to be able to detect and resolve forthcoming incidents or complications. Backup implants are mandatory. LEVEL OF EVIDENCE: IV, therapeutic case series. PMID- 17084300 TI - Hybrid femoral fixation of soft-tissue grafts in anterior cruciate ligament reconstruction using the EndoButton CL and bioabsorbable interference screws: a biomechanical study. AB - PURPOSE: The purpose of this study was to evaluate the effect of hybrid femoral fixation with bioabsorbable interference screws (BioRCI; Smith & Nephew Endoscopy, Andover, MA) and EndoButton CL (Smith & Nephew Endoscopy) fixation. METHODS: Biomechanical testing of 3 different fixation techniques was performed by use of porcine hind-limb distal femurs and mature bovine extremity common extensor tendons. Two independent testing sessions were examined. The first testing session (group A) compared femoral fixation via the EndoButton CL device (n = 6) with femoral fixation via the EndoButton CL device with the addition of a BioRCI screw (n = 6). The second testing session (group B) compared femoral fixation via BioRCI screws alone (n = 6) with femoral fixation via the EndoButton CL device with the addition of a BioRCI screw (n = 6). The femur-graft complex was cyclically loaded between 50 and 250 N at 1 Hz for 1,000 cycles. After cycling, the amount of graft slippage was determined by measuring the change in grip-to-grip distance. The complex was then loaded to failure at 1 mm/s, and the ultimate tensile strength, stiffness, and mode of failure were determined. RESULTS: In group A the addition of an interference screw to the EndoButton CL fixation increased the ultimate tensile strength (1,364.7 +/- 102.4 N for EndoButton CL alone v 1,449.3 +/- 94.4 N for combined technique, P = .035) and stiffness (195.5 +/- 12.1 N/mm for EndoButton CL alone v 307.3 +/- 54.9 N/mm for combined technique, P = .004) and decreased the amount of graft slippage (2.6 +/- 0.5 mm for EndoButton CL alone v 2.0 +/- 0.3 mm for combined technique, P = .017). In group B the addition of the EndoButton CL device to interference screw fixation significantly increased the ultimate tensile strength (643.5 +/- 148.4 N for BioRCI screws alone v 1,290.3 +/- 254.4 N for combined technique, P = .004) but had no effect on stiffness (315.7 +/- 38.9 N/mm for BioRCI screws alone v 341.5 +/- 64.0 N/mm for combined technique, P = .267) or graft slippage (2.7 +/- 1.0 mm for BioRCI screws alone v 2.0 +/- 0.6 mm for combined technique, P = .087). CONCLUSIONS: Our study shows that hybrid femoral fixation of double-looped gracilis-semitendinosus grafts via the EndoButton CL device and a bioabsorbable interference screw is stronger than interference or EndoButton CL fixation alone with respect to ultimate tensile strength, stiffness, and slippage. The addition of an interference screw to suspensory fixation via the EndoButton CL device increased the ultimate tensile strength from 1,360 N to 1,450 N, improved reconstruction stiffness from 200 N/mm to 300 N/mm, and decreased the amount of graft slippage resulting from cyclic loading from 2.6 mm to 2.0 mm. CLINICAL RELEVANCE: The hybrid fixation of the EndoButton CL device and an interference screw is a stronger and stiffer construct than either device alone and allows for aperture fixation, which may translate into better clinical results. PMID- 17084301 TI - Self-reported outcomes after anterior cruciate ligament reconstruction: an internal health locus of control score comparison. AB - PURPOSE: The purpose of this study was to evaluate the influence of high or low internal health locus of control (HLOC) scores on knee function and sports activity self-reports. METHODS: The Multidimensional HLOC Scale, the Knee Outcome Survey (KOS) Activities of Daily Living Scale (ADLS) and Sports Activity Scale (SAS), and the 2000 International Knee Documentation Committee (IKDC) Subjective Knee Evaluation and Current Health Assessment surveys were mailed to 335 subjects at a minimum of 2 years after anterior cruciate ligament reconstruction. RESULTS: Of the subjects, 198 returned completed surveys at 5.1 +/- 2.9 years after surgery. Compared with the group with low internal HLOC scores, the group with high internal HLOC scores had better KOS-ADLS scores (89.6 +/- 13.7 v 77.4 +/- 23.9, P < .0001), KOS-SAS scores (85.2 +/- 18 v 70 +/- 29, P < .0001), global ADLS ratings (90.7 +/- 12.8 v 77.4 +/- 23.9, P < .0001), global SAS ratings (81.8 +/- 20.6 v 70 +/- 29.4, P < .0001), 2000 IKDC Subjective Knee Evaluation scores (80.9 +/- 17.7 v 68.3 +/- 25.2, P < .0001), and 2000 IKDC Current Health Assessment scores for physical function (90.2 +/- 14 v 80.5 +/- 24.6, P = .001), social function (92.6 +/- 14.8 v 85.8 +/- 21.1, P = .014), bodily pain (80.7 +/- 17.7 v 68.7 +/- 24.6, P < .001), mental health (77.9 +/- 14.2 v 70.6 +/- 19.2, P = .005), and vitality (65.1 +/- 19 v 58.8 +/- 21, P = .04). Current knee function scores (8.7 +/- 1.8 v 7.1 +/- 2.9, P < .0001) and KOS-SAS sports activity level scores (chi(2) = 11.2, P = .01) were also higher in the group with high internal HLOC scores compared with the group with low scores. CONCLUSIONS: Patients with high internal HLOC scores were more satisfied with knee function. LEVEL OF EVIDENCE: Level IV, prognostic case series. PMID- 17084302 TI - Factors affecting the outcome of arthroscopy in medial-compartment osteoarthritis of the knee. AB - PURPOSE: This study was designed to obtain information on factors affecting the medium-term efficacy of arthroscopy (debridement or microfracturing of chondral defects) in patients with unicompartmental osteoarthritis (OA) of the knee. METHODS: A total of 156 patients (71 men and 85 women; mean age, 51.6 +/- 8.7 years [range, 37 to 69 years]) with isolated Kellgren-Lawrence grade 2 medial compartment knee OA underwent arthroscopy and were followed up. Patients with patellofemoral or lateral-compartment OA were excluded. The Knee Injury and Osteoarthritis Outcome Score was determined in all patients. The outcome was rated poor if this score was less than 114 points or if further surgery was required. Odds ratios (ORs) were calculated by use of multiple logistic regression adjusted for significantly associated factors. RESULTS: Follow-up was performed in 92.9% of the patients, at a mean of 49.2 +/- 2.1 months (range, 47 to 54 months). The outcome was poor in 104 patients (71.7%). There were no gender differences. Factors significantly associated with a poor outcome were a history of OA for greater than 24 months (OR, 3.6), obesity (OR, 8.8), smoking (OR, 3.1), medial tibial osteophytes (OR, 5.4), medial joint space width on standing radiographs of less than 5 mm (OR, 7.3), absence of effusion (OR, 6.5), absence of synovitis (OR, 6.1), presence of crystal deposits (OR, 4.3), deep tibial cartilage defect (OR, 12.5), and need for subtotal or total meniscectomy (OR, 2.2). Patients with more than 4 of these factors had significantly poorer outcomes. CONCLUSIONS: The medium-term outcome of arthroscopy in unicompartmental OA of the knee is poor in about 71.7% of the cases. In this study the outcome did not depend exclusively on the articular findings. Patient age was not associated with a poor outcome. However, a history of OA for more than 2 years, obesity, smoking, tibial osteophytes, and joint space narrowing of less 5 mm were associated with a poor outcome. Patients with 4 or more of these factors should be managed with treatment other than arthroscopy. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 17084303 TI - The posteromedial rim of the talar dome as the site for harvesting cartilage in the ankle: an anatomic study. AB - PURPOSE: The goal of this study was to determine whether a biopsy specimen of a fixed size can be harvested reliably and consistently by arthroscopy at the posteromedial rim of the talar dome. METHODS: A cartilage biopsy specimen was taken post mortem arthroscopically from the posteromedial rim of the talar dome in 20 ankles. We aimed to take a full-thickness biopsy specimen of 10 x 5 mm in size. The shape, length, width, position, and depth of the created defect were determined. Subsequently, 2 observers analyzed the biopsy sites twice. The same set of parameters and the surface area of the lesion were determined. Differences between aimed and observed sizes were studied. RESULTS: In all ankles the aimed biopsy site could be seen and reached. The observed mean size of the biopsy specimens, when compared with the aimed size, was only significantly different for the mediolateral size. A wide variation of surface area was found. Nearly all biopsy specimens started exactly at the posterior border of the dome, but they were somewhat more lateral than intended. In only half of the biopsy specimens was the shape linear or oval, the others being rounded or irregular. In case a pre-existing posteromedial cartilage lesion was present, its position did not coincide with that of the biopsy specimen. CONCLUSIONS: A limited cartilage biopsy specimen could be harvested reliably and consistently by arthroscopy at the posteromedial rim of the talar dome, although the size tended to be somewhat larger than we intended. Containment of a pre-existing posteromedial lesion was never jeopardized. CLINICAL RELEVANCE: A safe biopsy site for cartilage in the ankle eliminates donor-site morbidity in the knee and may improve the suitability and biology of the obtained chondrocytes for successful repair of symptomatic joint surface defects of the ankle. PMID- 17084304 TI - A new technique to improve tissue grip: "the lasso-loop stitch". AB - In the chain of reattachment of the rotator cuff, links extend from the tendon to the anchor within the bone. Passage of the suture through the tendon is noted at the start of the chain, but as shown before, this is most vulnerable, depending on the quality of the tendon. We present a new technique of suture placement designed to improve tissue grip: "The lasso-loop stitch." One side of the wire is passed through the edge of the tendon, which is ruptured by the wire in the middle-not at the end. The wire is not pulled through completely. Through this process, a loop is created at the upper side of the tendon. The end of the wire, at the same side used to make the loop, is passed through the loop with a grasp. The wire is passed through the loop, and the end of the wire is brought extra articularly. Through this technique, the reduction force of a suture is augmented, and a constricting factor is added. We have not changed the location of the anchors. So far, we have used the technique in rotator cuff repair, biceps tenodesis, and the Bankart procedure. PMID- 17084305 TI - Arthroscopic fixation of bursal-sided rotator cuff tears. AB - Subacromial decompression and debridement of partial-thickness bursal-sided rotator cuff tears are often reported with a high rate of unsatisfactory outcomes. We describe an arthroscopic procedure to repair partial-thickness bursal-sided rotator cuff tears without converting to a full-thickness tear in patients with a normal articular-sided rotator cuff and an A0B2 or A0B3 pattern of tear (minimum thickness of 25% to 75%). The articular side of the rotator cuff experiences greater tension than the bursal side of the cuff. As such, by leaving the articular footprint intact, we accomplish 3 goals: the intact articular fibers act as an internal splint to protect the bursal-sided repair, a wide and anatomic footprint is recreated, and we are able to minimize any length-tension mismatch because the tissue is not excessively lateralized with repair. After bursectomy and acromioplasty, the frayed edges of the bursal flap are gently debrided and the tuberosity is excoriated to bleeding bone. One or two bioabsorbable anchors are placed, and both sutures are placed through the full thickness of the rotator cuff (one anterior and one posterior) by use of a percutaneous suture lasso in this manner: (1) the lasso is passed through the full thickness of the cuff, and the nitinol wire is shuttled out of a single cannula along with the more medial of the suture limbs; (2) the nitinol wire is then pulled back out of the percutaneous portal along with the suture limb, with the suture being passed through the full thickness of the cuff; and (3) the procedure is repeated for the posterior limb of the suture after a lasso is again passed through the full thickness of the rotator cuff in a more posterior position. This subset of patients is treated with an aggressive rehabilitation protocol because the intact articular rotator cuff fibers act as an internal splint to protect the bursal repair. PMID- 17084306 TI - Arthroscopic placement of a modified Mason-Allen stitch. AB - Recent studies have shown all-arthroscopic rotator cuff repairs to have comparable clinical results to mini-open or open repairs. Previous drawbacks to arthroscopic repair have included not being able to place a modified Mason-Allen stitch with a suture anchor technique. We present a technique using the Arthrex Scorpion device (Arthrex, Naples, FL) to place a modified Mason-Allen stitch arthroscopically via a double-loaded FiberWire metal suture anchor (Arthrex). The Scorpion suture passer places a stitch from inferior to superior through the torn rotator cuff. The suture is grabbed and reloaded into the Scorpion device. The device is then turned upside down, and a horizontal stitch is placed from superior to inferior. The stitch is retrieved again and reloaded a third time. The last pass is placed so that the final stitch passes anterior to the inferior suture but beyond the previously placed horizontal mattress stitch. This effectively reproduces the described biomechanically superior modified Mason Allen stitch. PMID- 17084307 TI - Anatomic double bundle: a new concept in anterior cruciate ligament reconstruction using the quadriceps tendon. AB - Surgical procedures for double-bundle anterior cruciate ligament reconstruction, which currently use hamstring graft, have been described, but some concerns remain regarding graft fixation and the ability to obtain adequate bundle size. We report an original double-bundle anterior cruciate ligament reconstruction technique using a quadriceps tendon graft and a simplified outside-in femoral tunnel-drilling process. The graft consists of a patellar bone block with its attached tendon split into superior and inferior portions, which yields 2 bundles. The anteromedial tunnel is drilled from the outside through a small lateral incision by use of a guide. The posterolateral tunnel is made through the same incision with a specific guide engaged in the anteromedial tunnel. A single tibial tunnel is created. The graft is routed from the tibia to the femur with the bone block in the tibial tunnel and the 2 bundles in their respective femoral tunnels. After fixation of the bone block in the tibia, the 2 bundles are tensioned and secured separately in their femoral tunnels. PMID- 17084308 TI - Aperture fixation in arthroscopic anterior cruciate ligament double-bundle reconstruction. AB - The native anterior cruciate ligament (ACL) consists of 2 bundles, which have distinct biomechanical yet synergistic functions with respect to anterior tibial translation and combined rotatory loads. Traditionally, most ACL reconstruction techniques have primarily addressed the restoration of the anteromedial bundle, and less consideration was given to the posterolateral bundle. Recently, various ACL double-bundle reconstruction techniques have been described. With most of these techniques, however, an indirect extra-anatomic fixation far from the articular surface was performed. Because extra-anatomic fixation techniques, rather than aperture fixation techniques, are associated with graft tunnel motion, windshield wiper action, and suture stretch-out, concerns may arise regarding delayed biological incorporation, tunnel enlargement, and secondary rotational and anterior instability. We, therefore, present a novel arthroscopic technique that reapproximates the footprints of native ACL with the use of double strand semitendinosus and gracilis autografts for reconstruction of the anteromedial and posterolateral bundles, respectively. A separate femoral and tibial tunnel is drilled for each double-strand autograft. The femoral tunnel for the anteromedial bundle is drilled primarily through a transtibial technique, and the femoral tunnel for the posterolateral bundle is drilled via an accessory anteromedial portal with the use of a 4-mm offset drill guide in the anteroinferior aspect of the femoral tunnel for the anteromedial bundle. Bioabsorbable interference screws are used in aperture fixation for anatomic fixation of each bundle. This technique attempts to reproduce closely the native ligament and its biomechanical function. PMID- 17084309 TI - Femoral interference screw placement through the tibial tunnel: a novel method without graft damage. AB - A frequently encountered problem in endoscopic 1-incision anterior cruciate ligament (ACL) reconstruction is the difficulty involved in accurately inserting the femoral interference screw without significant and undesirable divergence between the screw and the graft when a femoral interference screw is fixed through the anteromedial portal. To minimize divergence, the authors demonstrated a modified, easy, and reproducible procedure that can be performed without causing graft injury or requiring special instrumentation. A ligament reconstruction route is created with the use of an ACL guide system. Lead graft sutures are pulled through the anteromedial portal by way of the femoral tunnel and out the anterolateral thigh first. The tendon graft is then inserted through the anteromedial portal and up into the femoral tunnel. A guidewire is introduced through the tibial tunnel into the femoral tunnel. An appropriately sized BioScrew (Linvatec, Largo, FL) is inserted, with the use of a guidewire inside the screw, through the tibial tunnel into the femoral tunnel. The graft is then retrieved through the anteromedial portal and is inserted through the tibial tunnel. Finally, the tendon graft in the tibial tunnel is similarly fixed with a BioScrew of the same size. Moreover, this novel approach is feasible for all tendon grafts (bone-patellar tendon-bone grafts, quadriceps tendon-patellar bone grafts, and hamstring tendon grafts). PMID- 17084310 TI - Arthroscopic reduction and repair of a locked posterior shoulder dislocation. AB - Locked posterior shoulder dislocations are an uncommon but difficult problem for the orthopaedic clinician. Furthermore, they are often missed on initial presentation, resulting in significant delays in treatment. Traditional treatment has involved formal open reduction, most commonly from an anterior approach, followed by transfer of the lesser tuberosity or subscapularis tendon into the anterior humeral head defect. We present the case of a patient with locked posterior shoulder dislocation, who was treated with arthroscopically assisted reduction followed by arthroscopic posterior stabilization. Use of this technique allows the surgeon to reduce the dislocation without performing an open arthrotomy, thereby decreasing the patient's overall morbidity. Furthermore, an arthroscopic technique used for stabilization allows visualization of the entire glenohumeral joint and enables the surgeon to directly address posterior disease, rather than compensating for the defect with an anteriorly based transfer. PMID- 17084311 TI - An unusual cause of knee pain 10 years after arthroscopy. AB - Arthroscopic surgery, although relatively minimally invasive, is associated with specific complications. Cases of instruments retained in the knee joint after arthroscopy have been reported. There have been reports of scalpel blades detaching into the joint during the approach that were immediately retrieved, with or without arthrotomy. We report the case of a scalpel blade retained in the knee joint for 10 years after arthroscopy. This case demonstrates the need for the surgeon and the whole operating room staff to be aware of the instrumentation and to systematically check the instruments at the beginning, middle, and end of the procedure. The significance of reporting this case is to make the orthopaedic community aware of possible complications that can occur even with relatively "low-risk" procedures. PMID- 17084312 TI - Protection of posterior structures during transtibial tunnel creation for posterior cruciate ligament reconstruction. PMID- 17084318 TI - Molecular diagnostics in sepsis: from bedside to bench. AB - BACKGROUND: Based on recent in vitro data, we tested the hypothesis that microarray expression profiles can be used to diagnose sepsis, distinguishing in vivo between sterile and infectious causes of systemic inflammation. STUDY DESIGN: Exploratory studies were conducted using spleens from septic patients and from mice with abdominal sepsis. Seven patients with sepsis after injury were identified retrospectively and compared with six injured patients. C57BL/6 male mice were subjected to cecal ligation and puncture, or to IP lipopolysaccharide. Control mice had sham laparotomy or injection of IP saline, respectively. A sepsis classification model was created and tested on blood samples from septic mice. RESULTS: Accuracy of sepsis prediction was obtained using cross-validation of gene expression data from 12 human spleen samples and from 16 mouse spleen samples. For blood studies, classifiers were constructed using data from a training data set of 26 microarrays. The error rate of the classifiers was estimated on seven de-identified microarrays, and then on a subsequent cross validation for all 33 blood microarrays. Estimates of classification accuracy of sepsis in human spleen were 67.1%; in mouse spleen, 96%; and in mouse blood, 94.4% (all estimates were based on nested cross-validation). Lists of genes with substantial changes in expression between study and control groups were used to identify nine mouse common inflammatory response genes, six of which were mapped into a single pathway using contemporary pathway analysis tools. CONCLUSIONS: Sepsis induces changes in mouse leukocyte gene expression that can be used to diagnose sepsis apart from systemic inflammation. PMID- 17084319 TI - Small rural hospitals and high-risk operations: how would regionalization affect surgical volume and hospital revenue? AB - BACKGROUND: Because higher hospital procedure volume is associated with better outcomes for many high-risk procedures, regionalization to higher-volume hospitals has been proposed as a way to improve quality of surgical care. The potential impact of such policies on small rural hospital volume and revenue is unknown. STUDY DESIGN: We identified all hospitalizations in small rural hospitals (less than 50 beds) in New York State from 1998 to 2001 that included an ICD-9 procedure code for 1 of 9 procedures for which there is a documented volume-outcomes association: abdominal aortic aneurysm repair, aortic-valve replacement, carotid endarterectomy, colectomy, coronary artery bypass, cystectomy, esophagectomy, pancreatectomy, or pulmonary resection. Revenue from these procedures was estimated using gross charges and payor-specific reimbursement rates. We then compared these estimates with total hospital inpatient revenue for each rural hospital. RESULTS: We identified 14 small rural hospitals where at least one of the nine procedures was performed. All included hospitalizations for colectomy. Aortic aneurysm repairs, cystectomies, and pancreatectomies were performed in three hospitals; carotid endarterectomy in two; and esophagectomy in one. In no hospitals were cardiac procedures or pulmonary resections performed. Estimated average contribution to hospital net revenue for all 9 procedures was approximately 2%, nearly all attributable to colectomy. CONCLUSIONS: If all aortic aneurysm repairs, major cardiothoracic procedures, carotid endarterectomies, cystectomies, and pancreatectomies in New York State were regionalized to higher-volume hospitals, no small rural hospitals would experience substantial impact in terms of rural hospital procedure volume and revenue. Even regionalization of colectomy would have a small impact on inpatient volume and revenue. PMID- 17084320 TI - Falls from height: spine, spine, spine! AB - BACKGROUND: Falls from a height are a major cause of morbidity and hospital cost. Spinal injury is frequently associated with falls from height, but reliable predictive factors have not been identified. Diagnostic evaluation of the spine is complex and debated. Our objective was to characterize spinal injury after falls from height and identify predictive factors of spinal injuries. STUDY DESIGN: Medical records of patients with falls from height>10 feet admitted in a Level I trauma center during a period of 66 months were reviewed. Univariate and multivariate analyses were performed to identify independent risk factors of spinal injuries. RESULTS: Of 414 patients, 127 (31%) suffered 277 spinal injuries. Multiple spinal injuries at different levels were found in 62 (49%) patients; in 19 (15%) spinal injuries were at noncontinuous levels. The only independent predictor of spinal injury was alcohol intoxication (odds ratio=3.305; 95% CI, 1.75-6.242; p<0.001) but the number of intoxicated patients was low and the predictive ability weak. Level of falls from height did not correlate with likelihood of spinal injury. Twenty-four of 107 (22%) patients with spinal injuries and a reliable clinical examination had no symptoms related to the spine; all but 2 had distracting injuries. CONCLUSIONS: Spinal injury is frequent among survivors of falls from height>10 feet. Because of the absence of reliable predictors of spinal injury, the possibility of multiple noncontinuous fractures, and the presence of distracting injuries clouding the clinical presentation, aggressive evaluation of the entire spine is warranted. PMID- 17084321 TI - Management and hospital outcomes of blunt renal artery injuries: analysis of 517 patients from the National Trauma Data Bank. AB - BACKGROUND: Blunt renal artery injuries are rare and no single trauma center can accumulate substantial experience for meaningful conclusions about optimal therapeutic strategies. The purpose of this study was to assess the incidence of renal artery injuries after different types of blunt trauma, and evaluate the current therapeutic approaches practiced by American trauma surgeons and the effect of various therapeutic modalities on hospital outcomes. STUDY DESIGN: This was a National Trauma Data Bank study including all blunt trauma admissions with renal artery injuries. Demographics, mechanism of injury, Injury Severity Score, Abbreviated Injury Score for each body area (head, chest, abdomen, extremities) injuries, type of management (nephrectomy, arterial reconstruction, or observation), time from admission to definitive treatment, and hospital outcomes (mortality, ICU, and hospital stay) were analyzed. Multiple and logistic regression analyses were used to examine the relationship between type of management and hospital outcomes. RESULTS: Of a total of 945,326 blunt trauma admissions, 517 patients (0.05%) had injuries to the renal artery. Of the 517 patients, the kidney was not explored in 376 (73%), 95 (18%) patients had immediate nephrectomy, and 45 (9%) patients underwent surgical revascularization. In 87 of 517 (17%) patients, renal artery injury was the only intraabdominal injury. Of the 87 patients with isolated renal artery injuries, 73 (84%) were observed, 7 (8%) underwent surgical revascularization, and 7 (8%) had early nephrectomy. Multiple regression analysis demonstrated that patients who had surgical revascularization had a considerably longer ICU and hospital stay than observed patients. Patients who had nephrectomy had a considerably longer hospital stay than observed patients. CONCLUSIONS: Blunt renal artery injury is rare. Nonoperative management should be considered as an acceptable therapeutic option. PMID- 17084322 TI - Identification of surgical complications and deaths: an assessment of the traditional surgical morbidity and mortality conference compared with the American College of Surgeons-National Surgical Quality Improvement Program. AB - BACKGROUND: Despite advances by surgeons in assessing quality and safety, the traditional surgical morbidity and mortality (M&M) conference has mostly remained unchallenged and unchanged. The goal of this study was to compare data as reported in a traditional M&M conference to data collected using the American College of Surgeons-National Surgical Quality Improvement Program (ACS-NSQIP) techniques. STUDY DESIGN: A retrospective study was performed comparing data from the M&M conference in a general surgery division, in which complications and deaths were identified by residents or attendings, to data compiled by a nationally audited nurse reviewer from the ACS-NSQIP from July 1, 2002, to June 30, 2003. RESULTS: Mortality rates calculated by traditional M&M conference (53 deaths in 5,905 patients), compared with the ACS-NSQIP nurse reviewer (28 deaths in 1,439 patients; 24% sample), were 0.9% versus 1.9%, respectively (p=0.001). Complication rates reported in M&M were 6.4% versus 28.9% ACS-NSQIP (p<0.0001). Subgroup analyses showed that mortality rates, as reported in conference, were substantially lower for both in-hospital and postdischarge patients, when compared with ACS-NSQIP. All subclassifications of complications, as presented in conference, were also lower, compared with ACS-NSQIP. CONCLUSIONS: Traditional surgical M&M reporting considerably underreports both in-hospital and postdischarge complications and deaths as compared with ACS-NSQIP. Approximately one of two deaths and three of four complications were not reported in the M&M conference at our institution. A Web-based reporting system based on an ACS-NSQIP platform was created to automate, facilitate, and standardize data on surgical morbidity and mortality. PMID- 17084323 TI - National Surgical Quality Improvement Program analysis of bariatric operations: modifiable risk factors contribute to bariatric surgical adverse outcomes. AB - BACKGROUND: The increase in obesity coupled with greater acceptance of the field of bariatric surgery has resulted in a substantial rise in the number of weight loss operations. Because obese individuals are at high risk for surgical complications, concern about the safety of bariatric procedures exists. Earlier investigations of the clinical features associated with surgical complications have produced conflicting results. We sought to identify risk factors for surgical complications in a large, nationally representative population of US veterans. STUDY DESIGN: We analyzed data on bariatric procedures performed at 12 Veterans' Affairs medical centers approved to perform weight-loss operations between 1998 and 2004. Detailed pre-, intra-, and postoperative information and longterm mortality data were prospectively collected using the National Surgical Quality Improvement Program methodology. We used multivariable logistic regression to identify clinical features associated with postoperative complications. RESULTS: Among 575 bariatric patients assessed between 1998 and 2004, 74% were men with a mean age of 51 years. Thirty-day mortality was 1.4%. Overall complication rate was 19.7%. Of those with complications, one-half were of considerable clinical importance, as they were associated with prolonged length of stay. Clinical features that were predictive of adverse events in our multivariable analyses were superobesity, weight>350 pounds, and smoking. A more than 20 pack-year history of smoking was also associated with difficulty in weaning from a ventilator postoperatively. CONCLUSIONS: We identified smoking and superobesity as preoperative risk factors associated with postoperative complications. Future studies should examine the effect of preoperative weight loss and smoking cessation on bariatric procedure outcomes. PMID- 17084324 TI - An institutional approach to maintenance of excellent outcomes for carotid endarterectomy in a setting with moderate-to-low procedure volume. AB - BACKGROUND: Institutions and surgeons with high procedure volumes have been reported to have lower morbidity and mortality rates for patients undergoing carotid endarterectomy. Demonstrating comparable results is essential for centers with moderate or low volume. If comparable results cannot be demonstrated, a low- to moderate-volume center should not perform the procedure. STUDY DESIGN: A prospective study of a program to achieve and sustain excellent outcomes after carotid endarterectomy was conducted at a single institution with low-to-moderate volumes. Results of this effort from January 1997 through December 2005 are reported. Key features of our approach include institutional control over which and how many surgeons can perform carotid endarterectomy. Surgeons must be experienced carotid surgeons and consistently perform more than 12 procedures annually and be continuously monitored. Surgical outcomes were independently audited through a mandated institutional carotid endarterectomy data registry. Surgeons with poor outcomes are barred from doing carotid endarterectomies; and annually updated outcomes data are posted on the Internet. RESULTS: This approach was used for 555 carotid endarterectomies in 503 patients. Our outcomes-with total death and disabling stroke rate of 1.6%-compare favorably with, and are not statistically different from, published benchmarks, despite volumes at our institution ranging from 44 to 81 patients annually and the participation of 8 surgeons during the study period. CONCLUSIONS: Surgeons should perform carotid endarterectomies only if excellent outcomes can be demonstrated. We conclude that by using an approach like ours, even institutions with moderate-to-low carotid endarterectomy volumes can achieve excellent outcomes. We propose that all institutions should assume responsibility for ensuring excellent carotid endarterectomy outcomes using a comprehensive outcomes-based approach with independent auditing similar to that presented here. PMID- 17084325 TI - Can induced anxiety from a negative earlier experience influence vascular surgeons' statistical decision-making? A randomized field experiment with an abdominal aortic aneurysm analog. AB - BACKGROUND: Increasing detection, new screening recommendations, and popular press attention contribute to the rising prevalence of asymptomatic abdominal aortic aneurysms (AAA). Evidence-based guidelines recommend the optimal time to operate is when the aneurysm is 5.5 cm in diameter. Smaller AAAs are periodically monitored with imaging. Recent events and emotional reactions to risk and uncertainty, including anxiety, can cause decision-making to diverge from cognitively based assessments. It is not known whether this applies to vascular surgeons making statistically-optimal, risky decisions. We tested whether an unexpected, recent negative event might influence vascular surgeons' decisions about a computer-simulation AAA-analog that includes statistical information. STUDY DESIGN: We performed a randomized, computer-based field experiment with evidenced-based statistical information readily available on bursting probabilities. Participants included vascular surgeons with AAA operative experience attending two vascular surgery conferences held in 2005 (n=81). The intervention was a randomly assigned, anxiety-inducing, bursting balloon versus a nonbursting balloon before a statistical decision-making computer simulation. The main outcomes measure was real-time prospective choice to opt out of expanding AAA simulation. A Cox proportional hazard model was used to assess the likelihood of opting out, while controlling for important covariates. RESULTS: The experimental group was more likely to opt out (hazard ratio: 3.32; 95% CI: 1.25 to 8.81), even after controlling for initial anxiety levels, risk attitudes, uncertainty attitudes, use of statistical information, surgical experience, and demographics. CONCLUSIONS: Experiencing a negative, potentially anxiety provoking, preceding event can influence decision-making, even among experienced vascular surgeons who have ready access to statistical risk information. PMID- 17084326 TI - Intraabdominal complications after lung transplantation. AB - BACKGROUND: Because more lung transplant recipients survive the perioperative period, nonpulmonary complications become a major source of morbidity and mortality. Of these, intraabdominal complications are of particular concern because of the potential need for surgical intervention. So appropriate management of these complications becomes paramount. STUDY DESIGN: We retrospectively reviewed 229 lung transplant recipients in a university medical center, between January 1997 and December 2004 developed in forty-seven patients. Abdominal complications. Detailed reviews of these patients' hospital charts were performed. Complications were categorized as early or late depending on if they occurred within 30 days of transplantation or later. The primary outcomes variable studied was mortality. RESULTS: Fifty-three surgical consultations for abdominal symptoms were requested in these 47 patients. Twenty-two of the 47 patients (47%) with intraabdominal complications required 24 operative interventions. Overall 5-year survival was substantially worse in patients with intraabdominal complications (34%) than in those without (62%, p=0.01). There was no marked difference in the 30-day mortality for patients experiencing early (27%, 4 of 15) versus late (24%, 9 of 38) complications. Mortality in patients with intraabdominal complications was lower among those treated operatively (n=2, 9%) compared with those treated nonoperatively (n=11, 44%, p=0.02). CONCLUSIONS: Mortality for patients with intraabdominal complications is high after lung transplantation. Operative intervention is well tolerated and associated with lower mortality. A high index of suspicion and timely operative intervention are necessary for the treatment of intraabdominal complications in lung transplant recipients. PMID- 17084327 TI - Surgical complications do not affect longterm survival after esophagectomy for carcinoma of the thoracic esophagus and cardia. AB - BACKGROUND: Surgical resection is the only real chance of cure for carcinoma of the esophagus and esophagogastric junction, although it carries considerable postoperative morbidity and mortality. The longterm prognosis for patients undergoing operation depends largely on the pathologic stage of the disease. The real impact of postoperative complications on survival is still under evaluation. STUDY DESIGN: A retrospective analysis was performed on patients with squamous cell carcinoma and adenocarcinoma of the thoracic esophagus and esophagogastric junction, undergoing surgical resection between January 1992 and December 2002. For the 522 patients considered for esophagogastroplasty, we analyzed comorbidities, preoperative staging, neoadjuvant treatments, surgical data, histopathology, postoperative surgical or medical complications, and survival. RESULTS: Surgical complications occurred in 85 of 522 patients (16.3%); their survival rate was entirely similar to that of the group of patients without surgical complications (p=0.9). The survival rate was worse for patients with concurrent surgical and medical complications. Analysis of the 99 patients (19%) who had only medical complications postoperatively revealed a survival rate comparable (p=0.9) with that of the 338 patients (63.7%) with an uneventful postoperative course. The median postoperative hospital stay was 14 days for all 522 patients, and 18 days for patients with medical or surgical postoperative complications. Multivariate analysis of the predictive factors showed that surgical complications do not affect longterm prognosis. CONCLUSIONS: Surgical complications have no negative impact on survival rates, which seem to depend exclusively on the pathologic stage of the tumor. PMID- 17084328 TI - Recovery from liver failure after hepatectomy for hepatocellular carcinoma in cirrhosis: meaning of the model for end-stage liver disease. AB - BACKGROUND: Hepatectomy for hepatocellular carcinoma in cirrhosis is followed by an impairment of liver function that can lead to patient death. The model for end stage liver disease (MELD) is considered an index of hepatic functional reserve, and its assessment on postoperative course may properly identify individuals at risk of liver failure. STUDY DESIGN: Two hundred hepatectomies for hepatocellular carcinoma in cirrhosis were reviewed. Irreversible postoperative liver failure was defined as an impairment of liver function after hepatectomy that led to patient death or required transplantation. The MELD scores at postoperative days (POD) 1, 3, 5, and 7 were calculated and kinetics of changes investigated with t test; logistic regression was applied to identify predictive variables of postoperative liver failure. RESULTS: Kinetics of postoperative MELD score showed an impairment of liver function between PODs 1 and 3; 185 patients in whom postoperative liver failure did not develop showed a considerable decrease in MELD score between PODs 3 and 5 (11.9+/-2.8 and 10.6+/-2.4, respectively, p<0.001). On the contrary, 15 patients, who experienced the event, showed an increase in MELD score between PODs 3 and 5 (18.2+/-3.9 and 18.3+/-3.6, respectively; p=0.845). Multivariate analysis showed preoperative MELD score (p<0.001), major hepatectomy (p=0.028), and MELD score increase between PODs 3 and 5 (p=0.011) as independent predictors of irreversible postoperative liver failure. Scores are reported as mean+/-SD. CONCLUSIONS: Recovery from liver impairment after hepatectomy for hepatocellular carcinoma in cirrhosis starts from POD 3; MELD scores increasing between PODs 3 and 5 may identify patients at risk of liver failure and represents the trigger for beginning intensive treatment or evaluating salvage transplantation. PMID- 17084329 TI - Outcomes after major hepatectomy in elderly patients. AB - BACKGROUND: We aimed to study the early and longterm outcomes of patients 70 years and older undergoing major liver resections, and compare the results with patients below the age of 70 years. STUDY DESIGN: All patients undergoing major liver resection (defined as three segments or more) from January 1993 to June 2004 were included. Patients were studied in two groups: 70 years of age and older (group E, elderly) and less than 70 years old (group Y, young). Early outcomes and longterm survival were analyzed. RESULTS: A total of 517 patients underwent major liver resection: group E, n=127; group Y, n=390 patients. There was no difference in operative mortality (group E, 7.9%; group Y, 5.4%; p=0.32) or postoperative morbidity (p=0.22) between the groups. Overall and disease-free survivals were not notably different for all patients (59% versus 57%, p=0.89; 60% versus 55%, p=0.28, respectively) or for a subgroup of patients with colorectal liver metastases (61% versus 55%, p=0.76; 60% versus 47%, p=0.07) in groups E versus Y, respectively. In multivariable analysis, American Society of Anesthesiologists grade 3 (p=0.024, hazard ratio [HR]=1.59, versus grade 1, 95% CI=1.06 to 2.39) and intraoperative transfusion>3 U (p<0.0005, HR=2.56, 95% CI=1.84 to 3.56) were predictors for overall survival. More than three tumors (p=0.025, HR=1.41, 95% CI=1.04 to 1.90) and redo resection (p=0.001, HR=2.80, 95% CI=1.51 to 5.19) were predictors of disease-free survival. CONCLUSIONS: Major liver resections can be safely performed in patients 70 years of age or older, with early results and survival similar to those in the younger than 70 age group. American Society of Anesthesiologists grade 3 and intraoperative transfusions>3 U were predictors for overall survival, and more than three tumors and redo resection were predictors for disease-free survival. PMID- 17084330 TI - Efforts to improve local control in rectal cancer compromise survival by the potential morbidity of optimal mesorectal excision. AB - BACKGROUND: The technique of total mesorectal excision (TME) increases the risk of anastomotic leakage. The impact of postoperative morbidity of TME on longterm survival has never been described. We retrospectively analyzed factors that might influence survival after TME for rectal cancer, including postoperative morbidity. STUDY DESIGN: From 1994 to 2001, 300 patients (192 men and 108 women; mean age, 64 years) had TME for rectal cancer. Preoperative radiotherapy was given in 202 patients. Age, gender, tumor height, size and circular invasion of the tumor, pathologic tumor and nodal status, distal and circumferential margins, number of lymph nodes analyzed, type of surgery, postoperative pelvic sepsis, preoperative radiotherapy, and adjuvant chemotherapy were examined; their association with overall and disease-free survival was evaluated by the log-rank test in univariate analysis and by multivariable Cox proportional hazards analysis. RESULTS: Postoperative morbidity was 38% (113 of 300 patients) and included 18% (54 of 300 patients) pelvic sepsis. The local recurrence rate was 6% (18 of 300 patients), and the distant metastasis rate was 24% (73 of 300 patients). Recurrence was three times more frequent distally than locally, including patients with pelvic sepsis The 5-year overall and disease-free survival rates were 72% and 60%, respectively. Independent predictors of overall survival were age older than 64 years (odds ration [OR]=2.19, 95% CI 1.32 to 4.17), pelvic sepsis (OR=2.06, 95% CI 1.10 to 3.87), circumferential surgical margin (OR=3.19, 95% CI 1.67 to 6.09), pathologic tumor (OR=2.69, 95% CI1.23 to 5.88), and nodal status (OR=3.18, 95% CI 1.79 to 5.64). Independent predictors of disease-free survival were pelvic sepsis (OR=2.17, 95% CI 1.31 to 3.58), circumferential surgical margin (OR=2.61, 95 CI 1.52 to 4.49), pathologic tumor (OR=1.82, 95% CI 1.04 to 3.20), and nodal status (OR=2.67, 95% CI 1.68 to 4.23). Patients with pelvic sepsis had a 5-year disease-free survival of 39% compared with 65% without pelvic sepsis (p<0.001). CONCLUSIONS: After TME for rectal cancer, pelvic sepsis is a common complication that is associated with increased risk of distant recurrence and decreased longterm survival. Efforts are necessary to decrease postoperative morbidity in surgical treatment of rectal cancer. PMID- 17084331 TI - Short-term impact of a laparoscopic "mini-residency" experience on postgraduate urologists' practice patterns. AB - BACKGROUND: To assist practicing urologists incorporate laparoscopic urology into their practice, a 5-day mini-residency (M-R) program with a mentor, preceptor, and proctor experience was established at the University of California, Irvine, and we report the initial results. STUDY DESIGN: Thirty-two urologists underwent laparoscopic ablative (n=17) or laparoscopic reconstructive (n=15) training, including inanimate model skills training, animal laboratory, and operating room observation. A questionnaire was mailed 1 to 15 months (mean, 8 months) after their M-R program, and responses were reviewed. RESULTS: A 100% response rate was achieved. The mean M-R participant age was 49 years (range 31 to 70 years). The majority of the participants (72%) had laparoscopic experience during residency training and had performed between 5 and 15 laparoscopic cases before attending the M-R program. Within 8 months after M-R, 26 participants (81%) were practicing laparoscopic surgery. Participants were performing laparoscopic radical nephrectomy (p=0.008), nephroureterectomy (p<0.0005), and pyeloplasty (p=0.008) at substantially higher rates after training. At the same time, fewer of the M-R participants were performing hand-assisted laparoscopic surgery after training (p=0.008) compared with before the M-R. Ninety-two percent of the participants indicated that they would recommend this training program to a colleague. CONCLUSIONS: A 5-day intensive laparoscopic ablative and reconstructive surgery course seems to encourage postgraduate urologists, already familiar with laparoscopy, to successfully expand the scope of their procedures to include more complex laparoscopic techniques such as nephrectomy, nephroureterectomy, and pyeloplasty into their clinical practice. PMID- 17084332 TI - Resection of neurogenic tumors in children: is thoracoscopy superior to thoracotomy? AB - BACKGROUND: Minimally invasive resection of solid tumors is controversial because of concerns of inadequate resection and local recurrence. Thoracoscopy has been used in the diagnosis of mediastinal tumors in children, but its role in resection is unproved. The purpose of this study was to compare thoracoscopic and open approaches to the resection of thoracic neurogenic tumors in children. STUDY DESIGN: The tumor registry of a regional children's hospital was queried to identify patients who underwent resection of neurogenic tumors over a 6-year period. Thoracoscopic and open groups were compared for demographic, operative, oncologic, and outcomes characteristics. RESULTS: Seventeen children underwent resection of mediastinal neurogenic tumors (10 thoracoscopic resections, 7 open resections). Mean age was 4.7 years (range 6 months to 12 years). The thoracoscopic and open groups showed no difference in operative time or blood loss. Tumors in the two groups were comparable in size (5.2+/-2.2 cm versus 5.7+/ 2.6 cm), histology, surgical margin, and stage. Hospital stay was shorter after thoracoscopic resection (1.9+/-0.7 days versus 4.1+/-2.5 days, p<0.05). There were no regional recurrences. Distant metastases developed in one patient in each group. Eight of 10 children with malignant tumors remain disease-free at an average of 25 months of followup (range 3 to 80 months). CONCLUSIONS: Thoracoscopic resection of neurogenic tumors achieved similar local control and disease-free survival when compared with open resection in this preliminary series. These results were accompanied by a shorter hospital stay. These findings suggest that thoracoscopic resection of neurogenic tumors in children may offer advantages to open resection and should be studied in the context of a large, cooperative trial. PMID- 17084333 TI - Nipple-sparing mastectomy for breast cancer and risk reduction: oncologic or technical problem? AB - BACKGROUND: We evaluated the risks and benefits of nipple-sparing mastectomy in a multiinstitutional experience in the settings of risk-reducing surgery and breast cancer treatment. STUDY DESIGN: We analyzed data on 123 patients who had undergone nipple-sparing mastectomy with breast reconstruction for prophylaxis (n=55), treatment of breast cancer (n=41), or both (n=27) at four large centers. RESULTS: Median patient age was 45 years (range 22 to 70 years). There were 192 procedures (69 bilateral, 54 unilateral). Forty-four patients had invasive cancer; 20 had ductal carcinoma in situ (DCIS); 4 had phyllodes tumor. In all of these patients, the nipple tissue was cancer free on pathologic review. Median followup was 24.6 months (range 2.0 to 570.4 months). Local recurrence developed in two patients: one had DCIS in the upper-outer quadrant, with 71.8 months of followup; the other's cancer was invasive, in the upper-outer quadrant, with 6 months of followup. Distant metastasis developed in a third patient, who died 50 months after the procedure. Breast cancer developed in two patients after prophylactic mastectomy: one in the upper-outer quadrant at 61.8 months; one in the axillary tail at 24.4 months. No patients had recurrences in the nipple areolar complex. Necrosis of the nipple was reported in 22 of 192 patients (11%) and it was judged minimal (less than one-third total skin of nipple) in 13 of 22 patients (59%). Overall cosmesis was judged by the patient and surgeon as good to excellent in the majority of patients. Level of satisfaction with cosmetic results was similar between prophylactic and treatment patients. CONCLUSIONS: The risk of local relapse was very low in our series of nipple-sparing mastectomies performed for DCIS or invasive cancer. Nipple-sparing mastectomy in the risk reducing and breast cancer-treatment settings may be feasible in selected patients and should be the subject of additional prospective clinical trials. PMID- 17084334 TI - Insular thyroid carcinoma: collective analysis of clinicohistologic prognostic factors and treatment effect with radioiodine or radiation therapy. AB - BACKGROUND: Insular thyroid carcinoma is a distinctive clinicopathologic entity with aggressive behavior and frequent lethal disease. But because it is rare, no single institute has sufficient experience with the clinicopathologic features and treatment effect of radioiodine. A literature review and collective analysis might facilitate patient management. METHODS: From January 1991 to December 2005, 9 patients with insular thyroid carcinoma were diagnosed in Taipei Veterans General Hospital. Clinicopathologic data were collected for analysis. The English literature about insular carcinoma was searched in Medline. Patients with detailed descriptions of treatment and followup were enrolled. Clinicopathologic features, rate of lymph node and distant metastasis, and prognostic factors were analyzed. RESULTS: Seventy-three of 486 patients collected from 23 articles with detailed description of treatment and followup were combined with our 9 patients for analysis, making a total of 82 patients in this article. The mean period of patient followup was 84.5 months. There were 26 (31.7%) men and 56 (68.3%) women. The rate of lymph node metastasis was 49%, and distant metastasis was 57.5%. Disease-specific death rate was 37.8%. The 5- and 10-year survival rates of insular carcinoma were 72.2% and 52%, respectively. Patient age older than 45 years (p=0.0004) and distant metastasis (p<0.0001) were poor prognostic factors. Use of radioiodine therapy (p=0.9789) and radiation therapy (p=0.2172) were not associated with prolonged survival. CONCLUSIONS: Patients with insular thyroid carcinoma have unfavorable outcomes. Postoperative radioiodine therapy or radiotherapy seemed not to be strongly associated with prolonged survival. PMID- 17084335 TI - Risk factors for perioperative complications in patients undergoing laparoscopic cholecystectomy: analysis of 22,953 consecutive cases from the Swiss Association of Laparoscopic and Thoracoscopic Surgery database. AB - BACKGROUND: Reliable risk factors for perioperative complications in patients undergoing laparoscopic cholecystectomy would be extremely useful to optimize the clinical management. This study aimed to determine risk factors that can be used for predicting perioperative complications. STUDY DESIGN: Possible risk factors for perioperative complications in patients undergoing laparoscopic cholecystectomy for acute and chronic cholecystitis were analyzed by a stepwise logistic regression model using data from the Swiss Association of Laparoscopic and Thoracoscopic Surgery (SALTS) database. RESULTS: A total of 22,953 patients with a mean (+/-SD) age of 54.5+/-16.1 years (range 17 to 89 years) and a male-to female ratio of 1:2, underwent elective (85%) and emergency (15%) laparoscopic cholecystectomy. Multivariable analysis showed that male gender (odds ratio [OR]=1.16; p<0.0001), duration of intervention (OR=1.68 per 30 minutes; p<0.0001), body weight (>90 kg versus<60 kg; OR=1.34; p<0.0001), and the surgeon's own experience (>100 versus 11 to 100 interventions; OR=1.36; p<0.0002) were independently associated with an increased intraoperative local complication rate. In addition, male gender (OR=1.21; p<0.02), age (OR=1.12 per 10 years; p<0.0001), intraoperative complications (OR=2.1; p<0.0001), conversion to open surgery (OR=1.25; p<0.01), American Society of Anesthesiologists risk score (ASA score III/IV versus I/II: OR=1.28; p<0.0005), body weight (<60 kg versus>90 kg; OR=1.53; p<0.007), emergency surgery (OR=1.36; p<0.003), and duration of surgery (OR=1.28 per 30 minutes; p<0.0001) were found to be associated with a higher incidence of postoperative local complications. Higher postoperative systemic complications were encountered with conversion (OR=1.5; p<0.0002), ASA score (III/IV versus I/II: OR=1.54; p<0.0001), emergency surgery (OR=1.41; p<0.001), and a prolonged intervention time (OR=1.16 per 30 minutes; p<0.0001). CONCLUSIONS: For patients undergoing laparoscopic cholecystectomy (LC), the risk of possible perioperative complications can be estimated based on patient characteristics (gender, age, ASA score, body weight), clinical findings (acute versus chronic cholecystitis), and the surgeon's own clinical practice with LC. So in the likelihood of a case being a "difficult cholecystectomy," an experienced surgeon should be involved both in the decision-making process and during the operation. If LC lasts longer than 2 hours, the cumulative risk for perioperative complications is four times higher compared with an intervention that lasts between 30 and 60 minutes, independent of the surgeon's personal skills with LC. PMID- 17084336 TI - Antibiotic prophylaxis for preventing wound infection after breast surgery: a systematic review and metaanalysis. PMID- 17084337 TI - Strategies to minimize postoperative deconditioning in elderly surgical patients. PMID- 17084338 TI - Medical considerations in older surgical patients. PMID- 17084339 TI - The elderly surgical patient and postoperative delirium. PMID- 17084340 TI - Parathyroid histopathology: is it of any value today? PMID- 17084341 TI - Application of rehabilitation ethics to a selected burn patient population's perspective. AB - BACKGROUND: Flame injury confers significant physical and psychologic stress on burn patients. Because most patients and their surrogates lack an understanding of burn injury, and these injuries preclude the meaningful exercise of patient autonomy, informed consent is a challenge for physicians. To better promote patient autonomy, this project collected patient perspectives on the proper duties of patients and physicians after severe burn injury and throughout the recovery phases. STUDY DESIGN: Ten survivors of severe burn injury were prospectively identified to represent different causes of injury, support systems, and socioeconomic backgrounds. Six persons participated (4 men, 2 women). Personal interviews with these individuals discussed their perspective and experience regarding physician and patient duties after severe burn injury as they relate to patient autonomy. RESULTS: All participants thought that informed consent was unrealistic at the time of their injury, but that this capacity developed over time as their understanding and level of functioning improved. In addition, all believed that the burn physicians' role was to do whatever was medically best for their patients in an emergency situation, but that this duty included the physician educating the patient or surrogates about what these treatments entail. CONCLUSIONS: Patient autonomy may be an unrealistic goal acutely for patients with severe burn injuries. Educational approaches to consent may facilitate patient autonomy, participation in decision making, and adherence to care plan over time. The ethical framework for this approach has been accepted in rehabilitation literature, but this is the first demonstration that relevant patient populations agree with this approach. PMID- 17084342 TI - Advanced Trauma Operative Management course: site and instructor selection and evaluation. PMID- 17084343 TI - Aortocaval fistula. PMID- 17084344 TI - Migratory gallstone causing duodenal obstruction. PMID- 17084345 TI - Spinal cord injuries. PMID- 17084347 TI - Triple-staple technique for low rectal anastomosis. PMID- 17084348 TI - Tourniquet use in the Civil War. PMID- 17084349 TI - Resolving polarized views: septin filaments do the twist. AB - Septins are a highly conserved family of filamentous GTPases required for cell division in a wide range of eukaryotic organisms. In a recent issue of Nature, Vrabioiu and Mitchison use polarized fluorescence microscopy to show that septin filaments undergo a highly orchestrated rotation, from a longitudinal to circumferential orientation, coincident with splitting of the septin ring during cytokinesis (Vrabioiu and Mitchison, 2006). PMID- 17084350 TI - Dissecting mitochondrial fusion. AB - Because mitochondria have outer and inner membranes, the fusion of two mitochondria requires the coordinated fusion of four lipid bilayers. Fusion of the outer membranes requires mitofusins. A new study published in Cell (Meeusen et al., 2006) shows that inner membrane fusion requires the dynamin-related protein Mgm1/OPA1. PMID- 17084351 TI - Myosin I and actin dynamics: the frogs weigh in. AB - In this issue of Developmental Cell, Sokac et al. (2006) describe an intriguing new role for an actin-based motor protein in restraining actin polymerization during endocytosis in Xenopus oocytes. PMID- 17084352 TI - SUMO: the glue that binds. AB - Two articles in a recent issue of Molecular Cell (Shen et al., 2006; Lin et al., 2006) demonstrate that noncovalent interactions between the SUMO moieties of SUMO modified PML, and SUMO binding motifs on PML and other PML nuclear-body associated proteins, affect the assembly of PML nuclear bodies and the recruitment of proteins in and out of these subnuclear structures. PMID- 17084353 TI - Imprinting weaves its web. AB - It has long been recognized that imprinted genes often act in common developmental or physiological pathways. A new knockout of the gene Zac1 reveals just how extensive the transcriptional network of imprinted genes may be. PMID- 17084354 TI - Catenins: keeping cells from getting their signals crossed. AB - Adherens junctions have been traditionally viewed as building blocks of tissue architecture. The foundations for this view began to change with the discovery that a central component of AJs, beta-catenin, can also function as a transcriptional cofactor in Wnt signaling. In recent years, conventional views have similarly been shaken about the other two major AJ catenins, alpha-catenin and p120-catenin. Catenins have emerged as molecular sensors that integrate cell cell junctions and cytoskeletal dynamics with signaling pathways that govern morphogenesis, tissue homeostasis, and even intercellular communication between different cell types within a tissue. These findings reveal novel aspects of AJ function in normal tissues and offer insights into how changes in AJs and their associated proteins and cytoskeletal dynamics impact wound-repair and cancer. PMID- 17084355 TI - Migration of zebrafish primordial germ cells: a role for myosin contraction and cytoplasmic flow. AB - The molecular and cellular mechanisms governing cell motility and directed migration in response to the chemokine SDF-1 are largely unknown. Here, we demonstrate that zebrafish primordial germ cells whose migration is guided by SDF 1 generate bleb-like protrusions that are powered by cytoplasmic flow. Protrusions are formed at sites of higher levels of free calcium where activation of myosin contraction occurs. Separation of the acto-myosin cortex from the plasma membrane at these sites is followed by a flow of cytoplasm into the forming bleb. We propose that polarized activation of the receptor CXCR4 leads to a rise in free calcium that in turn activates myosin contraction in the part of the cell responding to higher levels of the ligand SDF-1. The biased formation of new protrusions in a particular region of the cell in response to SDF-1 defines the leading edge and the direction of cell migration. PMID- 17084356 TI - Myosin-1c couples assembling actin to membranes to drive compensatory endocytosis. AB - Compensatory endocytosis follows regulated exocytosis in cells ranging from eggs to neurons, but the means by which it is accomplished are unclear. In Xenopus eggs, compensatory endocytosis is driven by dynamic coats of assembling actin that surround and compress exocytosing cortical granules (CGs). We have identified Xenopus laevis myosin-1c (XlMyo1c) as a myosin that is upregulated by polyadenylation during meiotic maturation, the developmental interval that prepares eggs for fertilization and regulated CG exocytosis. Upon calcium-induced exocytosis, XlMyo1c is recruited to exocytosing CG membranes where actin coats then assemble. When XlMyo1c function is disrupted, actin coats assemble, but dynamic actin filaments are uncoupled from the exocytosing CG membranes such that coats do not compress, and compensatory endocytosis fails. Remarkably, there is also an increase in polymerized actin at membranes throughout the cell. We conclude that XlMyo1c couples polymerizing actin to membranes and so mediates force production during compensatory endocytosis. PMID- 17084357 TI - The conserved c2 domain protein lethal (2) giant discs regulates protein trafficking in Drosophila. AB - Drosophila sensory organ precursor (SOP) cells undergo several rounds of asymmetric cell division to generate the four different cell types that make up external sensory organs. Establishment of different fates among daughter cells of the SOP relies on differential regulation of the Notch pathway. Here, we identify the protein Lethal (2) giant discs (Lgd) as a critical regulator of Notch signaling in the SOP lineage. We show that lgd encodes a conserved C2 domain protein that binds to phospholipids present on early endosomes. When Lgd function is compromised, Notch and other transmembrane proteins accumulate in enlarged early endosomal compartments. These enlarged endosomes are positive for Rab5 and Hrs, a protein involved in trafficking into the degradative pathway. Our experiments suggest that Lgd is a critical regulator of endocytosis that is not present in yeast and acts in the degradative pathway after Hrs. PMID- 17084358 TI - The Drosophila Notch inhibitor and tumor suppressor gene lethal (2) giant discs encodes a conserved regulator of endosomal trafficking. AB - Notch signaling is involved in many developmental and pathological processes, and its activity must be precisely controlled in order to prevent aberrant development and disease. We have previously shown that the tumor suppressor gene lethal (2) giant discs (lgd) is required to prevent ectopic activation of Notch in developmental processes in Drosophila. Here we show that lgd is required in all imaginal disc cells to suppress the activity of the Notch pathway. lgd encodes a member of a poorly characterized protein family present in all animals, which includes a member that is involved in an inheritable form of mental retardation in humans. Our analysis reveals that Lgd is required for endosomal trafficking of Notch and other proteins. In the absence of Lgd, Notch is activated in a ligand-independent manner in probably all imaginal disc cells in an endosomal compartment downstream of the block in hrs mutants. PMID- 17084359 TI - Phosphatidylinositol 4-phosphate formation at ER exit sites regulates ER export. AB - The mechanisms that regulate endoplasmic reticulum (ER) exit-site (ERES) assembly and COPII-mediated ER export are currently unknown. We analyzed the role of phosphatidylinositols (PtdIns) in regulating ER export. Utilizing pleckstrin homology domains and a PtdIns phosphatase to specifically sequester or reduce phosphorylated PtdIns levels, we found that PtdIns 4-phosphate (PtsIns4P) is required to promote COPII-mediated ER export. Biochemical and morphological in vitro analysis revealed dynamic and localized PtsIns4P formation at ERES. PtdIns4P was utilized to support Sar1-induced proliferation and constriction of ERES membranes. PtdIns4P also assisted in Sar1-induced COPII nucleation at ERES. Therefore, localized dynamic remodeling of PtdIns marks ERES membranes to regulate COPII-mediated ER export. PMID- 17084360 TI - Frodo links Dishevelled to the p120-catenin/Kaiso pathway: distinct catenin subfamilies promote Wnt signals. AB - p120-catenin is an Arm repeat protein that interacts with varied components such as cadherin, small G proteins, kinases, and the Kaiso transcriptional repressor. Despite recent advances in understanding the roles that p120-catenin and Kaiso play in downstream modulation of Wnt/beta-catenin signaling, the identity of the upstream regulators of the p120-catenin/Kaiso pathway have remained unclear. Here, we find that p120-catenin binds Frodo, which itself interacts with the Wnt pathway protein Dishevelled (Dsh). In Xenopus laevis, we demonstrate that Wnt signals result in Frodo-mediated stabilization of p120-catenin, which, in turn, promotes Kaiso sequestration or removal from the nucleus. Our results point to Dsh and Frodo as upstream regulators of the p120-catenin/Kaiso signaling pathway. Importantly, this suggests that Wnt signals acting through Dsh regulate the stability of p120-catenin in addition to that of beta-catenin, and that each catenin promotes its respective signal in parallel to regulate distinct, as well as shared, direct downstream gene targets. PMID- 17084361 TI - SoxD proteins influence multiple stages of oligodendrocyte development and modulate SoxE protein function. AB - The myelin-forming oligodendrocytes are an excellent model to study transcriptional regulation of specification events, lineage progression, and terminal differentiation in the central nervous system. Here, we show that the group D Sox transcription factors Sox5 and Sox6 jointly and cell-autonomously regulate several stages of oligodendrocyte development in the mouse spinal cord. They repress specification and terminal differentiation and influence migration patterns. As a consequence, oligodendrocyte precursors and terminally differentiating oligodendrocytes appear precociously in spinal cords deficient for both Sox proteins. Sox5 and Sox6 have opposite functions than the group E Sox proteins Sox9 and Sox10, which promote oligodendrocyte specification and terminal differentiation. Both genetic as well as molecular evidence suggests that Sox5 and Sox6 directly interfere with the function of group E Sox proteins. Our studies reveal a complex regulatory network between different groups of Sox proteins that is essential for proper progression of oligodendrocyte development. PMID- 17084362 TI - Zac1 regulates an imprinted gene network critically involved in the control of embryonic growth. AB - Genomic imprinting is an epigenetic mechanism of regulation that restrains the expression of a small subset of mammalian genes to one parental allele. The reason for the targeting of these approximately 80 genes by imprinting remains uncertain. We show that inactivation of the maternally repressed Zac1 transcription factor results in intrauterine growth restriction, altered bone formation, and neonatal lethality. A meta-analysis of microarray data reveals that Zac1 is a member of a network of coregulated genes comprising other imprinted genes involved in the control of embryonic growth. Zac1 alters the expression of several of these imprinted genes, including Igf2, H19, Cdkn1c, and Dlk1, and it directly regulates the Igf2/H19 locus through binding to a shared enhancer. Accordingly, these data identify a network of imprinted genes, including Zac1, which controls embryonic growth and which may be the basis for the implementation of a common mechanism of gene regulation during mammalian evolution. PMID- 17084363 TI - Multipotent flk-1+ cardiovascular progenitor cells give rise to the cardiomyocyte, endothelial, and vascular smooth muscle lineages. AB - Cell-tracing studies in the mouse indicate that the cardiac lineage arises from a population that expresses the vascular endothelial growth factor receptor 2 (VEGFR2, Flk-1), suggesting that it may develop from a progenitor with vascular potential. Using the embryonic stem (ES) cell differentiation model, we have identified a cardiovascular progenitor based on the temporal expression of the primitive streak (PS) marker brachyury and Flk-1. Comparable progenitors could also be isolated from head-fold stage embryos. When cultured with cytokines known to function during cardiogenesis, individual cardiovascular progenitors generated colonies that displayed cardiomyocyte, endothelial, and vascular smooth muscle (VSM) potential. Isolation and characterization of this previously unidentified population suggests that the mammalian cardiovascular system develops from multipotential progenitors. PMID- 17084364 TI - The C. elegans sex-determining GLI protein TRA-1A is regulated by sex-specific proteolysis. AB - TRA-1A is the sole representative in Caenorhabditis elegans of the Gli transcription factor family. Its activity is required to specify all somatic female cell fates in XX hermaphrodites. We have found that TRA-1 protein levels are much higher in hermaphrodites than in males, and that the difference is attributable to the predominance in hermaphrodites of C-terminally truncated isoforms that are nearly undetectable in males. Our results support a model in which TRA-1A is negatively regulated by male-specific proteolysis that depends upon specific TRA-1A protein sequences and upon the activity of the fem genes. C terminally truncated TRA-1 isoforms are stable and can inappropriately feminize XO males, suggesting that they escape this negative regulation. Thus, although C. elegans appears to lack a Hedgehog-signaling pathway, our results indicate that proteolytic processing and degradation of Gli family transcription factors, commonly seen during Hedgehog signaling in other organisms, also control C. elegans sex determination. PMID- 17084365 TI - Regulation of mitotic chromosome cohesion by Haspin and Aurora B. AB - In vertebrate mitosis, cohesion between sister chromatids is lost in two stages. In prophase and prometaphase, cohesin release from chromosome arms occurs under the control of Polo-like kinase 1 and Aurora B, while Shugoshin is thought to prevent removal of centromeric cohesin until anaphase. The regulatory enzymes that act to sustain centromeric cohesion are incompletely described, however. Haspin/Gsg2 is a histone H3 threonine-3 kinase required for normal mitosis. We report here that both H3 threonine-3 phosphorylation and cohesin are located at inner centromeres. Haspin depletion disrupts cohesin binding and sister chromatid association in mitosis, preventing normal chromosome alignment and activating the spindle assembly checkpoint, leading to arrest in a prometaphase-like state. Overexpression of Haspin hinders cohesin release and stabilizes arm cohesion. We conclude that Haspin is required to maintain centromeric cohesion during mitosis. We also suggest that Aurora B regulates cohesin removal through its effect on the localization of Shugoshin. PMID- 17084366 TI - Role of naturally arising regulatory T cells in hematopoietic cell transplantation. AB - Naturally arising CD4(+)CD25(+) regulatory T cells (Tregs) have the potential to suppress aberrant immune responses and to regulate peripheral T-cell homeostasis. In murine models of bone marrow transplantation, Tregs promote donor bone marrow engraftment and decrease the incidence and severity of graft-versus-host-disease without abrogating the beneficial graft-versus-tumor immunologic effect. These findings, in concert with observations that Tregs in mice and humans share phenotypic and functional characteristics, have led to active investigations into the use of these cells to decrease complications associated with human hematopoietic cell transplantation. Early human studies suggest that an imbalance of Tregs and effector T cells may contribute to the development of graft-versus host-disease. However, the mechanisms of immunoregulation, in particular the allorecognition properties of Tregs, their effects on and interaction with other immune cells, and their sites of suppressive activity, are not well understood. In this review, we discuss the current knowledge of Treg biology and the potential therapeutic strategies and barriers of Treg immunotherapy in human hematopoietic cell transplantation. PMID- 17084367 TI - Early vaccination with tumor-lysate-pulsed dendritic cells after allogeneic bone marrow transplantation has antitumor effects. AB - Allogeneic bone marrow transplantation (BMT) remains the primary treatment for many hematologic malignancies but has had limited success against solid tumors. The antitumor activity of this treatment approach involves the tumoricidal activity of chemoradiation and the additive graft-versus-tumor activity of donor T cells. However, even with current protocols, some tumors develop resistance and become unresponsive to current therapeutic regimens. To address the problem of resistance and lack of solid tumor activity in allogeneic BMT, we undertook experiments to determine whether the graft-versus-tumor activity of donor T cells could be enhanced in the period immediately after allogeneic BMT with tumor lysate-pulsed dendritic cell (DC) vaccines. Using the B16 melanoma model, we found that the treatment of 6-day tumors with allogeneic BMT and 3 weekly vaccinations of tumor lysate-pulsed DCs starting 3 days after BMT had a significant effect on the growth of murine flank melanomas. This effect was tumor specific and occurred in the absence of full immune reconstitution as measured by donor T cell engraftment and cytotoxic T lymphocyte activity. In addition, DC vaccinations did not appear to exacerbate graft-versus-host disease. These experiments support the feasibility of DC vaccine strategies in the setting of allogeneic BMT. PMID- 17084368 TI - Clinically relevant expansion of hematopoietic stem cells with conserved function in a single-use, closed-system bioprocess. AB - The clinical potential of umbilical cord blood-derived stem and progenitor cells has been demonstrated in various animal and human transplantation studies. However, the need for increased numbers of appropriate umbilical cord blood derived cells continues to limit the development and success of these therapies. Ex vivo expansion has been widely studied as a method to overcome this limitation. We describe the use of a clinically relevant single-use, closed system bioprocess capable of generating greater numbers of hematopoietic stem and progenitor cells that maintain in vivo and in vitro developmental potential. In addition to expanded numbers of CD34+ cells, CD34(+)CD38(-) cells, colony-forming cells, and long-term culture-initiating cells, the bioprocess generated > or =3.3 fold more long-term nonobese diabetic/severe combined immunodeficient repopulating cells (quantitatively determined using limiting dilution analysis) than present at input. Interestingly, these cells were also capable of multilineage engraftment and were shown to maintain their engraftment potency on a per long-term nonobese diabetic/severe combined immunodeficient repopulating cell basis compared with input noncultured cells. The developmental capacity of bioprocess-generated cells was further demonstrated by their ability to repopulate secondary nonobese diabetic/severe combined immunodeficient recipients. In vitro lineage analysis confirmed that bioprocess-generated cells could differentiate into myeloid and natural killer, B, and T cell lymphoid lineages. This in-depth analysis describes a bioprocess that generates human hematopoietic stem and progenitor cells with conserved hematopoietic activity, establishes analysis criteria for in vitro hematopoietic stem cell expansion studies, and serves as a foundation to test the therapeutic utility of cultured hematopoietic stem cells in large animals and humans. PMID- 17084369 TI - Predictors of improved progression-free survival after nonmyeloablative allogeneic stem cell transplantation for advanced chronic lymphocytic leukemia. AB - Although chronic lymphocytic leukemia (CLL) remains an incurable disease with standard chemotherapy, the appropriate role and timing of transplantation are unclear. In this analysis, we report the outcomes of 46 patients with advanced CLL who underwent nonmyeloablative stem cell transplantation (NST) from HLA matched unrelated (67%) or related (33%) donors. Fludarabine (30 mg/m2 x 4) and low-dose intravenous busulfan (0.8 mg/kg/day x 4) were used for conditioning. The 2-year overall survival (OS) and progression-free survival (PFS) rates in this refractory patient population were 54% and 34%, respectively, with a median follow-up of 20 months. The primary cause of treatment failure was relapse, with a 2-year cumulative incidence of 48%. High hematopoietic donor chimerism > or = 75% at day +30 was a significant predictor of 2-year PFS (47% vs 11%; P = .03). In multivariate analysis, chemotherapy-refractory disease at transplantation was associated with a 3.2-fold risk of progression (P = .01) and a 4.6-fold risk of death (P = .02). Increasing number of previous therapies and increasing bone marrow involvement were also associated with decreased PFS and OS. These results suggest that NST using fludarabine and low-dose intravenous busulfan is a reasonable treatment option for patients with advanced CLL, but that NST earlier in the disease course will likely be needed to achieve long-term disease control in a high proportion of patients. PMID- 17084370 TI - Long-term results of autologous stem cell transplantation for primary refractory or relapsed Hodgkin's lymphoma. AB - Autologous hematopoietic stem cell transplantation (ASCT) has become standard therapy for primary refractory (PR REF) or relapsed (REL) Hodgkin's lymphoma (HL); however, more than half of these patients eventually relapse and die of their disease. We studied long-term outcomes and evaluated factors influencing progression-free survival (PFS) in 141 patients with PR REF or REL HL who underwent ASCT between 1985 and 2003. Median age at ASCT was 30 years (range, 7 60 years); 21 patients had PR REF, and 120 had REL HL. With a median follow-up of 6.3 years (range, 1-20 years), the probability of PFS at 5 and 10 years was 48% (95% confidence interval [CI], 39%-57%) and 45% (95% CI, 36%-54%) and that of overall survival (OS) was 53% (95% CI, 44%-62%) and 47% (95% CI, 37%-57%), respectively. Transplant-related mortality at 100 days was 1.4%. Among 45 5- to 20-year survivors, no late relapses of HL were observed. In multivariate analysis, 3 factors were independently predictive of poor PFS: chemoresistant disease (relative risk [RR], 2.9; 95% CI, 1.7-5.0), B-symptoms at pretransplantation relapse (RR, 2.1; 95% CI, 1.3-3.4), and presence of residual disease at the time of transplantation (RR, 2.3; 95% CI, 1.1-4.8). Patients with 0 or 1 of these 3 adverse factors (low-risk disease) had a 5-year PFS of 67% (95% CI, 55%-79%) compared with 37% (95% CI, 22%-52%) in those with 2 factors (intermediate-risk group) and 9% (95% CI, 0-20%) in those with all 3 factors (high-risk group) (P < .001). The rates of OS at 5 years were 71% (95% CI, 60% 82%), 49% (95% CI, 33%-65%) and 13% (95% CI, 0-27%) in the 3 groups, respectively (P < .001). ASCT is associated with durable PFS in appropriately selected patients with PR REF or REL HL. Using a simple prognostic model, we can identify patients with high-risk disease who have predictably unfavorable outcome after ASCT and require novel therapeutic approaches. A risk-adapted approach should be followed in determining treatment options for patients with PR REF and REL HL. PMID- 17084371 TI - Unmanipulated HLA 2-3 antigen-mismatched (haploidentical) stem cell transplantation using nonmyeloablative conditioning. AB - The major problems in human leukocyte antigen (HLA)-mismatched stem cell transplantation (SCT) are graft failure and graft-versus-host disease (GVHD). Less-intensive regimens should be associated with a lower release of inflammatory cytokines and possibly less GVHD. The objective of this study was to investigate whether HLA-haploidentical SCT can be performed using nonmyeloablative conditioning and pharmacologic GVHD prophylaxis, including glucocorticoids. Using conditioning consisting of fludarabine, busulfan, and anti-T-lymphocyte globulin and GVHD prophylaxis consisting of tacrolimus and methylprednisolone (1 mg/kg/day), 26 patients who had hematologic malignancies in an advanced stage or with a poor prognosis underwent transplantation using peripheral blood stem cells from an HLA-haploidentical donor (2-3 antigen mismatches in the graft-versus-host [GVH] direction) without T-cell depletion. All patients except for 1 achieved donor-type engraftment. Rapid hematologic engraftment was achieved (neutrophils > 0.5 x 10(9)/L on day 12 and platelets > 20 x 10(9)/L on day 12), with full donor chimerism achieved by day 14. Fifteen patients did not develop acute GVHD clinically, and only 5 patients developed grade II GVHD. The recovery of CD4+ T cells was delayed compared with that of CD8+ T cells. Sixteen of the 26 patients are alive in complete remission. Four died of transplantation-related causes, and 6 died of progressive disease. These data suggest that nonmyeloablative conditioning, GVHD prophylaxis consisting of tacrolimus and methylprednisolone, and early therapeutic intervention for the GVH reaction allow stable engraftment and effectively suppress GVHD in HLA 2-3 antigen-mismatched SCT. PMID- 17084372 TI - Is high-dose chemotherapy after primary chemotherapy a therapeutic option for patients with primary mediastinal nonseminomatous germ cell tumor? AB - Patients with primary mediastinal nonseminomatous germ cell tumors have a poor prognosis, with a 5-year overall survival of nearly 50%. We investigated the feasibility and activity of early high-dose chemotherapy (HDCT) in these patients. After conventional induction chemotherapy, patients underwent a single shot of HDCT consisting of carboplatin, etoposide, and cyclophosphamide, followed by peripheral blood progenitor cell support. Twenty-one patients were considered for treatment with HDCT. Median age was 29 years (range, 19-55 years). Eight (38%) patients had lung metastases. After primary chemotherapy, 7 patients achieved complete remission, 4 achieved partial remission with negative marker, 1 achieved partial remission with positive marker, 2 had stable disease, and 7 progressive disease. Twelve patients were not treated with HDCT due to progressive disease and poor physical conditions. No HDCT-related deaths or irreversible organ toxicities were observed. Residual surgery after HDCT was performed in 4 patients and resulted in 3 pathologic complete remissions. With a median follow-up of 52 months (range, 15-71 months) in 9 patients treated with HDCT, 8 have been continuously free of disease. Of 12 patients who did not receive HDCT, 0 was alive at 2 years from diagnosis. A single course of HDCT after induction chemotherapy appeared to be inapplicable in most of our patients, mainly due to early progressive disease. These data should be considered in the analysis of retrospective series and in the design of new prospective trials with HDCT in these patients. Earlier HDCT administration followed by residual surgery should be considered for further investigation. PMID- 17084373 TI - Reconstitution of T cell subset repertoire diversity following multiple antigen mismatched bone marrow transplantation. PMID- 17084374 TI - Postexposure prophylaxis against varicella zoster virus infection among hematopoietic stem cell transplant recipients. PMID- 17084375 TI - Postcolumn nucleic acid intercalation for the fluorescent detection of nucleic acids using ion pair reverse phase high-performance liquid chromatography. AB - Here we report a simple and effective procedure enabling the fluorescent detection of nucleic acids following the rapid, high-resolution separation using ion pair reverse phase chromatography. This approach uses postcolumn nucleic acid intercalation of fluorescent dyes with subsequent fluorescent detection, demonstrating more than a 1000-fold increase in sensitivity in the detection of nucleic acids when compared with traditional UV detection. Moreover, a wide range of intercalating dyes can be incorporated, including those known to disrupt the structure of the nucleic acids, thereby enabling the sensitive detection of DNA and RNA with no adverse effect on resolution of the nucleic acids during ion pair reverse phase chromatography. In addition, such approaches allow one to readily distinguish single-stranded DNA from double-stranded DNA following their separation using ion pair reverse phase high-performance liquid chromatography. PMID- 17084376 TI - Fatty acid transport in articular cartilage. AB - Articular cartilage extracellular matrix imposes a significant transport barrier to albumin, the principal carrier of fatty acids. It has not been previously established whether it also influences the transport of fatty acids important for chondrocyte metabolism. Albumin was labelled with rhodamine-maleimide and bound to NBD-labelled lauric acid. Plugs of fresh equine metacarpal-phalangeal cartilage and subchondral bone were incubated with the complex at 4 degrees C for 2-160 h. The fluorophore distribution was quantified using quantitative microscopy in histological sections. The fluorescence intensity of both fluorophores fell steeply over 300 microm below the articular surface and remained relatively uniform through the mid zone but the ratio of lauric acid to albumin was higher than in the incubation medium. The effective diffusivity of lauric acid in the mid zone was (2.2+/-0.7) x 10(-12) m2 s(-1) (n = 33), higher than that of the carrier albumin, suggesting dissociation in the surface layer. Lauric acid accumulated reversibly at the tidemark. PMID- 17084377 TI - Folding and assembly pathways of co-chaperonin proteins 10: Origin of bacterial thermostability. AB - To compare folding/assembly processes of heptameric co-chaperonin proteins 10 (cpn10) from different species and search for the origin of thermostability in hyper-thermostable Aquifex aeolicus cpn10 (Aacpn10), we have studied two bacterial variants-Aacpn10 and Escherichia coli cpn10 (GroES)-and compared the results to data on Homo sapiens cpn10 (hmcpn10). Equilibrium denaturation of GroES by urea, guanidine hydrochloride (GuHCl) and temperature results in coupled heptamer-to-monomer transitions in all cases. This is similar to the behavior of Aacpn10 but differs from hmcpn10 denaturation in urea. Time-resolved experiments reveal that GroES unfolds before heptamer dissociation, whereas refolding/reassembly begins with folding of individual monomers; these assemble in a slower step. The sequential folding/assembly mechanism for GroES is rather similar to that observed for Aacpn10 but contradicts the parallel paths of hmcpn10. We reveal that Aacpn10's stability profile is shifted upwards, broadened, and also moved horizontally to higher temperatures, as compared to that of GroES. PMID- 17084378 TI - Characterization of the calpain/calpastatin system in human hemopoietic cell lines. AB - As previously suggested by PCR analysis [R. DeTullio, R. Stifanese, F. Salamino, S. Pontremoli, E. Melloni, Characterization of a new p94-like calpain form in human lymphocytes, Biochem. J. 375 (2003) 689-696], a p94-like calpain was now established to be present in six different human cells resembling the various peripheral blood cell types. This protease resulted to be the predominant calpain isoforms whereas the conventional mu- and m-calpains are also expressed although at lower or almost undetectable amounts. The p94-like calpain has been identified by a specific mAb and displays unique features such as: Ca2+ requirement for half maximum activity around 30 microM; no autolytic conversion to a low Ca2+ requiring form and lower sensitivity to calpastatin inhibition. Following cell stimulation, the p94-like calpain undergoes inactivation, a process indicating that the protease is activated and participates in the cell responses to stimuli. The involvement of this protease isoform in immunocompetent cell activation is further supported by its partial recruitment on plasma membranes, the site of action of the conventional calpain forms. The amount of calpain translocated to the membranes correlates to the level of calpastatin which has been shown to control this process through the formation of a complex with calpain, which maintains the protease in the cytosol. These results provide new information on the calpain/calpastatin system expressed in immunocompetent cells and on the functional relationship between the p94-like calpain and the biological function of these cells. PMID- 17084379 TI - Down-regulation of PPARgamma2-induced adipogenesis by PEGylated conjugated linoleic acid as the pro-drug: Attenuation of lipid accumulation and reduction of apoptosis. AB - This study is designed to evaluate whether the PEGylated conjugated linoleic acid (PCLA) as the pro-drug can have favorable stability, bioavailability, and anti adipogenic activity in 3T3-L1 cells for anti-obesity when compared with conjugated linoleic acid (CLA) itself. The CLA was simply coupled to poly(ethylene glycol) (PEG) at the melting state without solvents or catalysts through ester linkages between the carboxylic group of CLA and the hydroxyl group of PEG. To confirm of PCLA as the pro-drug, CLA release from PCLA was investigated by using high-performance liquid chromatographic (HPLC), showing that CLA release from PCLA was almost 90% in a nearly continuous fashion over the next 75h. Apoptosis was promoted by both CLA- and PCLA-treatments with increasing concentrations. However, the level of cell apoptosis induced by PCLA was lower than that induced by CLA owing to the biocompatible and hydrophilic properties of PEG. Moreover, the PCLA decreased glycerol-3-phosphate dehydrogenase (GPDH) activity in 3T3-L1 cells by acting upon major adipocyte marker proteins such as PPARgamma2, C/EBPalpha, and aP2 modulators. Furthermore, either CLA or PCLA stimulated basal, but not isoproterenol-sensitive, lipolysis in our cell model, suggesting that both CLA and PCLA may stimulate lipolysis via hormone sensitive lipase (HSL)-independent mechanisms. These results suggest that the PCLA may prove to be a stable pro-drug to control the deposition of fat in the human body, and that the anti-adipogenic effect of the PCLA on 3T3-L1 cells will offer a challenging approach for anti-obesity. PMID- 17084380 TI - Anxiety sensitivity within the anxiety disorders: disorder-specific sensitivities and depression comorbidity. AB - The tendency to perceive anxious states as aversive and harmful is hypothesized to confer vulnerability to the development of anxiety disorders. The most commonly used measure of anxiety sensitivity, the Anxiety Sensitivity Index [ASI; Reiss, S., Peterson, R.A., Gursky, D.M., & McNally R.J. (1986). Anxiety sensitivity, anxiety frequency, and the prediction of fearfulness. Behavior Research and Therapy, 24, 1-8], is composed of multiple lower-order factors, assessing fear of physical symptoms, fear of publicly observable anxious symptoms, and fear of cognitive dyscontrol. This study examined the convergent validity of the lower-order anxiety sensitivity dimensions in DSM-IV diagnosed anxiety disorders. Participants with primary diagnoses of panic disorder with agoraphobia, social phobia, and generalized anxiety disorder (GAD) completed the ASI and measures of anxiety and depression severity. Support was found for the convergent validity of all ASI dimensions in reference to thematically related anxiety disorders and in the identification of patients presenting with and without secondary major depressive disorder (MDD). The ASI-fear of cognitive dyscontrol dimension displayed strong and nonredundant associations with GAD, dimensional depression scores, and secondary diagnoses of MDD. The conceptual implications of the shared importance of fear of cognitive dyscontrol in GAD and MDD are discussed. PMID- 17084381 TI - Regulatory role of neuron-restrictive silencing factor in expression of TRPC1. AB - Neuron-restrictive silencer factor (NRSF) binds its consensus element to repress the transcription of various genes. The dominant-negative form (dnNRSF) has a hypertrophic effect on cardiogenesis through an unidentified mechanism. We examined the involvement of transient receptor potential (TRP) channel proteins, using transgenic mice overexpressing dnNRSF (dnNRSF mice). Electrophoretic mobility-shift assays revealed an interaction between NRSF and a neuron restrictive silencer element-like sequence in intron 4 of TRPC1 genomic DNA. According to RT-PCR and Western analyses, TRPC1 was up-regulated in dnNRSF mouse heart. Transient overexpression of TRPC1 in HEK 293T cells increased the activity of the nuclear factor in activated T cells (NFAT) promoter and stimulated store operated Ca(2+) channel (SOCC)-mediated Ca(2+) entry. Transfection of TRPC1 into primary cardiomyocytes increased NFAT activity, indicating a major role for TRPC1 in NFAT activation. Our findings strongly suggest that NRSF regulates TRP1 gene expression and causes changes in the levels of calcium entry through SOCCs. PMID- 17084382 TI - Cholesteryl ester hydroperoxides increase macrophage CD36 gene expression via PPARalpha. AB - The uptake of oxidized LDL by macrophages is a key event in the development of atherosclerosis. The scavenger receptor CD36 is one major receptor that internalizes oxidized LDL. In differentiated human macrophages, we compared the regulation of CD36 expression by copper-oxidized LDL or their products. Only oxidized derivatives of cholesteryl ester (CEOOH) increased the amount of CD36 mRNA (2.5-fold). Both oxidized LDL and CEOOH treatment increased two to fourfold the transcription of promoters containing peroxisome-proliferator-activated receptor responsive elements (PPRE) in the presence of PPARalpha or gamma. Electrophoretic-mobility-shift-assays with nuclear extracts prepared from macrophages treated by either oxidized LDL or CEOOH showed increased binding of PPARalpha to the CD36 gene promoter PPRE. In conclusion, CEOOH present in oxidized LDL increase CD36 gene expression in a pathway involving PPARalpha. PMID- 17084383 TI - CASK associates with glutamate receptor interacting protein and signaling molecules. AB - CASK has been implicated in synaptic protein targeting, synaptic organization, and transcriptional regulation. Here, three more CASK associated proteins, GRIP1, PKCepsilon, and RGS4, were initially identified by immunoprecipitation and mass analysis, and confirmed by immunoprecipitation-immunoblotting assay using rat brain extract. Via the interaction with GRIP1, GluR2/3 was also co immunoprecipitated by CASK antibody from rat brain. The PDZ and SH3-GK domains of CASK were demonstrated as the associated domains for GRIP1 and PKCepsilon, respectively. The associations between CASK, PKCepsilon, and RGS4 were up regulated in the adult brain compared with postnatal day 11 rat brain. In contrast, the associations of CASK with Mint1, GRIP1, and GluR2/3 were down regulated in the adult brain. These results suggest that CASK protein complex is developmentally regulated by unknown signals. In conclusion, our study suggests that the CASK protein complex not only functions as a scaffold but also recruits signaling molecules and may contribute to signal transduction. PMID- 17084384 TI - Allele-specific chromatin remodeling of the tumor necrosis factor-alpha promoter. AB - The -863 C/A polymorphism in the tumor necrosis factor-alpha (TNF-alpha) promoter has been suggested to influence TNF-alpha expression. Here we elucidated the molecular mechanisms underlying the allele-specific regulation of TNF-alpha gene expression under basal and LPS-stimulated conditions in THP-1 cells and in human primary macrophages. We show that the binding of two NF-kappaB complexes, the p50/p50 homodimer and the p50/p65 heterodimer, was upregulated upon LPS stimulation. Both complexes bound to the C-allele whereas the A-allele only bound the p50/p65 complex. Two DNase I hypersensitive sites appeared in the TNF-alpha promoter after LPS stimulation of THP-1 cells. DNase I hypersensitivity of the TNF-alpha promoter was also analyzed in human monocytes prepared from individuals of different -863C/A genotype. Hypersensitivity was increased in the promoter harboring the mutant A-allele, particularly after LPS stimulation. In summary, binding of transcription factor NF-kappaB to the TNF-alpha promoter is associated with allele-specific remodeling of chromatin structure. PMID- 17084385 TI - Pioglitazone acutely influences glucose-sensitive insulin secretion in normal and diabetic human islets. AB - We have studied acute effects of the PPARgamma agonist pioglitazone in vitro on human islets from both non-diabetic and type 2 diabetic subjects. In 5 mM glucose, pioglitazone caused a transient increase in insulin secretion in non diabetic, but not diabetic, islets. Continuous presence of the drug suppressed insulin release in both non-diabetic and diabetic islets. In islets from non diabetic subjects, both high glucose and tolbutamide-stimulated insulin secretion was inhibited by pioglitazone. When islets were continuously perifused with 5 mM glucose, short-term pretreatment with pioglitazone caused approximately 2-fold increase in insulin secretion after drug withdrawal. Pioglitazone pretreatment of diabetic islets restored their glucose sensitivity. Examination of cytosolic free Ca(2+) concentration ([Ca(2+)](i)) in non-diabetic islets revealed slight Ca(2+) transient by pioglitazone at 3 mM glucose with no significant changes at high glucose. Our data suggest that short-term pretreatment with pioglitazone primes both healthy and diabetic human islets for enhanced glucose-sensitive insulin secretion. PMID- 17084386 TI - Characterization of pericentrin isoforms in vivo. AB - Pericentrin was first identified as a mouse centrosomal protein and is now referred to as pericentrin A. A larger homologous protein in humans with a C terminal calmodulin-binding domain was later identified as pericentrin B. Pericentrin has been shown to be one of the key components in ciliogenesis, but in vivo pericentrin products have remained ambiguous. Here we characterized pericentrin isoforms in mice. Two pericentrin transcripts of 9.5 and 6.9 kb were recognized on the mouse tissue Northern blots, while a cRNA probe for a 5' terminal sequence shared by pericentrin A and B failed to hybridize to the 6.9-kb message. Two pericentrin cDNAs were identified, which encoded pericentrin B and a novel isoform, pericentrin S, sharing with pericentrin B a C-terminal calmodulin binding motif. Three pericentrin proteins of 360, 255, and 250 kDa revealed by immunoprecipitation analysis were thought to correspond to pericentrin B, pericentrin S, and an unknown N-terminal product. PMID- 17084387 TI - Differentiation stage-specific analysis of gene function with inducible short hair-pin RNA in differentiating embryonic stem cells. AB - Cell differentiation is regulated by spatial and temporal coordination of gene expressions. Previously, we have established an embryonic stem (ES) cell differentiation system that can trace early cardiovascular developmental process in vitro. Here we show that tetracycline-induced short hair-pin RNA (shRNA) expression in differentiating ES cells successfully suppressed stage-specific genes for differentiation and modified cell fates. We established ES cell lines carrying shRNA gene driven by tRNA(val) promoter with tetracycline operator sequences (tet-ON system). When expression of vascular endothelial growth factor receptor-2 (VEGFR2) gene, a vascular progenitor and mesoderm marker and an essential gene for endothelial cell (EC) differentiation, was suppressed by shRNA in early ES cell differentiation, appearance of VEGFR2(+) mesoderm cells was substantially reduced. Suppression of VEGFR2 expression at mesoderm stage almost completely inhibited EC differentiation from VEGFR2(+) mesoderm cells. This novel experimental system, thus, can selectively determine stage-specific roles of genes in differentiation in vitro. PMID- 17084388 TI - Osmotic tolerance limits and effects of cryoprotectants on the motility, plasma membrane integrity and acrosomal integrity of rat sperm. AB - Osmotic stress is an important factor that can result in cell damage during cryopreservation. The objectives of this study were to determine: (1) isosmotic sperm cell volume; (2) osmotically inactive volume; (3) osmotic tolerance limits of rat sperm; and (4) the effects of addition and removal of glycerol (Gly), ethylene glycol (EG), propylene glycol (PG) or dimethyl sulfoxide (Me(2)SO) on rat sperm function. Sperm from Fischer 344 and Sprague-Dawley rats were used in this study. An electronic particle counter was used to measure the cell volume of rat sperm. Computer-assisted sperm motility analysis and flow-cytometric analysis were used to assess sperm motility, plasma membrane and acrosomal integrity. The isosmotic sperm cell volumes of the two strains were 37.0+/-0.1 and 36.2+/-0.2 microm(3), respectively. Rat sperm behaved as linear osmometers from 260 to 450 mOsm, and the osmotically inactive sperm volumes of the two strains were 79.8+/ 1.5% and 81.4+/-2.2%, respectively. Rat sperm have very limited osmotic tolerances. The sperm motility and the sperm plasma membranes of both strains were sensitive to anisosmotic treatments, but the acrosomes of both strains were more sensitive to hyposmotic than hyperosmotic conditions. The one-step addition and removal of Me(2)SO showed the most deleterious effect on rat sperm motility, plasma membrane integrity, and acrosomal integrity among the four cryoprotectants. These data characterizing rat sperm osmotic behavior, osmotic and cryoprotectant tolerance will be used to design cryopreservation protocols for rat sperm. PMID- 17084389 TI - Catalytic competence of the Ras-GEF domain of hSos1 requires intra-REM domain interactions mediated by phenylalanine 577. AB - The Ras-specific guanine nucleotide exchange region of hSos1 consists of two consecutive domains: the catalytic core (residues 742-1024) contains all residues binding to Ras, including the catalytic hairpin, and an upstream REM domain (residues 553-741), so called because it contains an evolutionary conserved Ras Exchange Motif (REM). We functionally define the boundaries of the REM domain through a combination of in vivo and in vitro assays. We show that an intra-REM domain interaction, mediated by phenylalanine 577, is required to allow interaction of the REM domain with the catalytic core, constraining it in the active conformation. PMID- 17084390 TI - Localization and in vitro binding studies suggest that the cytoplasmic/nuclear tobacco lectin can interact in situ with high-mannose and complex N-glycans. AB - The possible in vivo interaction of the Nicotiana tabacum agglutinin (Nictaba) with endogenous glycoproteins was corroborated using a combination of confocal/electron microscopy of an EGFP-Nictaba fusion protein expressed in tobacco Bright Yellow-2 (BY-2) cells and biochemical analyses. In vitro binding studies demonstrated that the expressed EGFP-Nictaba possesses carbohydrate binding activity. Microscopic analyses confirmed the previously reported cytoplasmic/nuclear location of Nictaba in jasmonate-treated tobacco leaves and provided evidence for the involvement of a nuclear localization signal-dependent transport mechanism. In addition, it became evident that the lectin is not uniformly distributed over the nucleus and the cytoplasm of BY-2 cells. Far Western blot analysis of extracts from whole BY-2 cells and purified nuclei revealed that Nictaba interacts in a glycan inhibitable way with numerous proteins including many nuclear proteins. Enzymatic deglycosylation with PNGase F indicated that the observed interaction depends on the presence of N-glycans. Glycan array screening, which showed that Nictaba exhibits a strong affinity for high-mannose and complex N-glycans, provided a reasonable explanation for this observation. The cytoplasmic/nuclear localization of a plant lectin that has a high affinity for high-mannose and complex N-glycans and specifically interacts with conspecific glycoproteins suggests that N-glycosylated proteins might be more important in the cytoplasm and nucleus than is currently believed. PMID- 17084391 TI - Protein dolichylation in Plasmodium falciparum. AB - We performed reverse-phase thin-layer chromatography and reverse-phase high performance liquid chromatography (RP-HPLC) analysis of polyisoprenoids released by sulfonium-salt cleavage with methyl iodide from Plasmodium falciparum proteins labeled with [3H]FPP or [3H]GGPP and showed that a dolichol of 11 isoprene units is bound to 21-28-kDa protein clusters from trophozoite and schizont stages. The dolichol structure was confirmed by electrospray-ionization mass spectrometry analysis. Treatment with protein synthesis inhibitors and RP-HPLC analysis of the proteolytic digestion products from parasite proteins labeled with [35S]cysteine and [3H]FPP showed that the attachment of dolichol to protein is a post translational event and probably occurs via a covalent bond to cysteine residues. PMID- 17084392 TI - The evolution of putative starch-binding domains. AB - The present bioinformatics analysis was focused on the starch-binding domains (SBDs) and SBD-like motifs sequentially related to carbohydrate-binding module (CBM) families CBM20 and CBM21. Originally, these SBDs were known from microbial amylases only. At present homologous starch- and glycogen-binding domains (or putative SBD sequences) have been recognised in various plant and animal proteins. The sequence comparison clearly showed that the SBD-like sequences in genethonin-1, starch synthase III and glucan branching enzyme should possess the real SBD function since the two tryptophans (or at least two aromatics) of the typical starch-binding site 1 are conserved in their sequences. The same should apply also for the sequences corresponding with the so-called KIS-domain of plant AKINbetagamma protein that is a homologue of the animal AMP-activated protein kinase (AMPK). The evolutionary tree classified the compared SBDs into three distinct groups: (i) the family CBM20 (the motifs from genethonins, laforins, starch excess 4 protein, beta-subunits of the animal AMPK and all plant and yeast homologues, and eventually from amylopullulanases); (ii) the family CBM21 (the motifs from regulatory subunits of protein phosphatase 1 together with those from starch synthase III); and (iii) the (CBM20+CBM21)-related group (the motifs from the pullulanase subfamily consisting of pullulanase, branching enzyme, isoamylase and maltooligosyl trehalohydrolase). PMID- 17084393 TI - Effects of gonadotropin-releasing hormone agonist/recombinant follicle stimulating hormone versus gonadotropin-releasing hormone antagonist/recombinant follicle-stimulating hormone on follicular fluid levels of adhesion molecules during in vitro fertilization. AB - OBJECTIVE: To compare the effects of GnRH-agonist/recombinant rFSH versus GnRH antagonist/recombinant FSH stimulation on follicular fluid levels of soluble intercellular adhesion molecule (sICAM)-1 and vascular cell adhesion molecule-1 (sVCAM-1) during in vitro fertilization (IVF). DESIGN: Prospective, randomized study. SETTING: University hospital. PATIENT(S): Seventy-three women underwent IVF. INTERVENTION(S): GnRH-agonist/rFSH or GnRH-antagonist/rFSH administration and collection of follicular fluid from 3 small (11-14 mm in diameter) and 3 large (18-21 mm in diameter) follicles on the day of oocyte retrieval. MAIN OUTCOME MEASURE(S): Follicular fluid levels of sICAM-1 and sVCAM-1 and intrafollicular estradiol and progesterone were also measured. RESULT(S): Women who underwent GnRH-agonist/rFSH showed higher concentrations of sICAM-1 in both small and large follicles were compared with patients who received GnRH antagonist/rFSH treatment; follicular fluid levels of sVCAM-1 were similar between the 2 stimulation protocols. Content of sICAM-1 in small and large follicles positively correlated with the number of follicles of > or =15 mm and the number of oocytes that were retrieved in both study groups. Concentrations of follicular fluid sVCAM-1 and progesterone were higher in large than in small follicles and were correlated positively to each other in both follicular classes. CONCLUSION(S): In IVF, GnRH-agonist/rFSH is associated with higher follicular fluid levels of sICAM-1 compared with GnRH-antagonist/rFSH regimen. Intrafollicular sICAM-1 content may predict ovarian response, and sVCAM-1 appears as an indicator of the degree of follicular luteinization. PMID- 17084394 TI - Mutational analysis of betaglycan/TGF-betaRIII in premature ovarian failure. AB - Targeted mutational screening of the inhibin-binding domain of betaglycan (exons 11 and 12) demonstrated an increased occurrence of the exon 12 194969T-->C variant in POF patients (27/54; 50%) compared to controls (8/41; 19.5%; P=.002); however, no gene variations were identified in exon 11. PMID- 17084395 TI - Measuring estradiol levels after human chorionic gonadotropin administration for in vitro fertilization is not clinically useful. AB - A retrospective analysis of 844 IVF-ET cycles demonstrated that changes in E(2) levels after administration of hCG do not influence fertilization, implantation, pregnancy, or live-birth rates. In vitro fertilization cycles with a declining E(2) level have comparable success to those with no change or increasing E(2) levels and should proceed to oocyte retrieval. PMID- 17084396 TI - Use of a gestational carrier for a patient with recurrent adverse pregnancy outcomes from early onset severe pre-eclampsia. AB - OBJECTIVE: To describe the first reported case of gestational carrier treatment to prevent severe early onset pre-eclampsia. DESIGN: Case report. SETTING: A university-based reproductive endocrinology and infertility clinic and a tertiary care hospital. PATIENT(S): A 29-year-old woman and her husband with three consecutive pregnancies complicated by early onset severe pre-eclampsia causing fetal demises at 22 and 24 weeks gestation; a neonatal death at 25 weeks gestation; and life-threatening maternal hemolysis, elevated liver enzymes, and low platelets. INTERVENTION(S): An IVF procedure in the patient using her husband's sperm with the transfer of two embryos to a friend who offered to be a gestational carrier. MAIN OUTCOME MEASURE(S): Successful IVF cycle in the patient and uncomplicated pregnancy and delivery in the gestational carrier. RESULT(S): The gestational carrier achieved a pregnancy and progressed without complications to delivery of a healthy, 3.2-kg infant at 39 weeks gestation. CONCLUSION(S): The use of a gestational carrier deserves consideration as a treatment option in patients with poor reproductive histories because of early onset severe pre eclampsia and hemolysis, elevated liver enzymes, and low platelets syndrome. This experience also suggests that development of pre-eclampsia may be in large part maternally rather than embryologically or paternally driven. PMID- 17084397 TI - Increased levels of air pollution and a decrease in the human and mouse male-to female ratio in Sao Paulo, Brazil. AB - A significant negative association (R(2) = 0.7642; P=.013) between particulate matter and secondary sex ratio was found when evaluating people in the city of Sao Paulo, Brazil. An animal model with male mice raised in nonfiltered open-top chambers showed a significant reduction in the secondary sex ratio (P=.041), suggesting that ambient air pollution may interfere with sex distribution by altering the X:Y sperm proportion in pollution-exposed males. PMID- 17084398 TI - Conjoined twins in a trichorionic quadruplet pregnancy after intracytoplasmic sperm injection and quarter laser-assisted zona thinning. AB - OBJECTIVE: To describe the occurrence and management of conjoined twins (CTs) in a trichorionic quadruplet pregnancy after the transfer of three embryos obtained by intracytoplasmic sperm injection (ICSI) that had undergone assisted zona hatching (AZH) using a noncontact laser. DESIGN: Case report. SETTING: Private and public IVF centers. PATIENT(S): A 38-year-old nulliparous woman. INTERVENTION(S): IVF, diagnosis by two-dimensional, three-dimensional, and four dimensional ultrasound, selective termination at 12 weeks gestation. MAIN OUTCOME MEASURE(S): Clinical follow-up and delivery. RESULT(S): After diagnosis, counselling, and selective termination, the subsequent prenatal course was uneventful, and two healthy boys were delivered by elective cesarean section at 38 weeks gestation. CONCLUSION(S): Monozygotic twinning (MZT) and CTs could complicate multifetal pregnancies obtained by assisted reproduction techniques. The relationship between CTs, IVF, and AZH should be better studied to fully understand the mechanisms that lead to MZT (with the possible role of AZH) and CTs in MZT, in particular. Three-dimensional and four-dimensional ultrasound improve prenatal assessment of the condition. Selective embryo reduction is a safe procedure for the management of these pregnancies. PMID- 17084399 TI - Ovarian cryopreservation with transposition of a contralateral ovary: a combined approach for fertility preservation in women receiving pelvic radiation. AB - OBJECTIVE: To describe a technique of fertility preservation by ovarian transposition combined with ovarian cryopreservation in the setting of oncologic pelvic radiation for a reproductive age woman. DESIGN: Case report. SETTING: IVF center at a tertiary care hospital. PATIENT(S): Thirty-year-old nulligravid women who previously underwent lumbar spinal cord tumor debulking requested fertility preservation before pelvic radiation. INTERVENTION(S): Laparoscopic ovarian transposition with cryopreservation of the contralateral ovary. MAIN OUTCOME MEASURE(S): New technique in fertility preservation. RESULT(S): Laparoscopic surgery was used to evaluate, select, and remove a single ovary that then underwent cryopreservation. Transposition of the remaining ovary was subsequently successfully performed, placing it of out of the pelvis. CONCLUSION(S): The combination of ovarian cryopreservation and ovarian transposition may maximize future fertility options for women facing pelvic irradiation. This combined approach should be included among the options offered to reproductive age women before pelvic radiation. PMID- 17084400 TI - Tumor necrosis factor-alpha regulates vascular endothelial growth factor secretion by human oviductal epithelial cells and stromal fibroblasts. AB - Tumor necrosis factor-alpha stimulates the secretion of vascular endothelial growth factor by cultured human oviductal epithelial cells and stromal fibroblasts. Tumor necrosis factor-alpha-stimulated vascular endothelial growth factor secretion by these cells may control oviductal fluid secretion by regulating vascular permeability. PMID- 17084401 TI - Copeptin, derived from isotocin precursor, is a probable prolactin releasing factor in carp. AB - Control of prolactin (PRL) release is of crucial importance for the multiple functions exerted by PRL in vertebrates. Recently identified hypothalamic PRL releasing peptides displayed additional neuromodulatory activities and in fish only few could be detected close to lactotrophs. Here we describe the C-terminal peptide processed from the carp isotocin precursor as probable physiologically relevant regulator of PRL release in carp. The amino acid sequence derived from the complete isotocin precursor gene of Cyprinus carpio, predicted a C-terminal peptide uncleaved between the neurophysin (Np) and copeptin (Cp) domain. Accordingly, antibodies against synthetic Np- and Cp-specific oligopeptides both immunodetected a 13kDa protein (cNpCp) in total pituitary proteins and showed abundant immunoreaction in hypothalamic axons in direct contact with lactotrophs in the rostral pars distalis of carp pituitary gland sections. Finally, incubation of cultured carp pituitary explants with purified carp cNpCp resulted in a potent stimulation of PRL release. PMID- 17084402 TI - Adiponectin is associated with improvement of endothelial function after rosiglitazone treatment in non-diabetic individuals with metabolic syndrome. AB - OBJECTIVE: The risks of metabolic syndrome (MetSyn) rely on the interaction between insulin resistance, metabolic abnormalities, inflammation and vascular dysfunction. Insulin sensitizers counteract some of these abnormalities. The objective of this study was to evaluate the effects of rosiglitazone (ROSI) on vascular reactivity, adipokines and inflammatory markers in a group of non diabetic subjects with MetSyn. METHODS AND RESULTS: Thirty subjects with NCEP ATPIII criteria for MetSyn and eight healthy subjects were studied at baseline and 18 subjects with MetSyn were treated with ROSI 8 mg/day for 24 weeks. Venous occlusion plethysmography exams before and during intra-arterial infusions of acetylcholine and sodium nitroprusside were performed to assess endothelial dependent and independent vasodilation. Forearm blood flow (FBF) and vascular resistance (VR) responses were analyzed. Treatment with ROSI improved endothelial function (235% increment in FBF; p<0.01 and 56% decrement in VR; p=0.01), adiponectin (7.3[3.6-17.9] versus 37.9[19.3-42.4]; p<0.01), HOMA-IR (3.5+/-1.2 versus 2.7+/-1.6; p<0.05), C-reactive protein (CRP) (1.05[0.57-2.07] versus 0.3[0.2-0.8]; p<0.01) and fibrinogen (3.1+/-0.73 versus 2.62+/-0.64; p<0.05) levels. The difference between groups on endothelial-dependent vasodilation, adiponectin and CRP levels disappeared after treatment and the improvement of endothelial function was associated with the increment of adiponectin levels. CONCLUSION: ROSI treatment restored endothelial function in MetSyn subjects, probably through an adiponectin-mediated process. PMID- 17084403 TI - Comments on "Stair climbing is more critical than walking in pre-clinical assessment of primary stability in cementless THA in vitro" by Jean-Pierre Kassi, Markus O. Heller, Ulrich Stoeckle, Carsten Perka, Georg N. Duda, Published on J. Biomechanics 2005; 38: 1143-1154. PMID- 17084404 TI - Chondroitin sulfate reduces the friction coefficient of articular cartilage. AB - The objective of this study was to investigate the effect of chondroitin sulfate (CS)-C on the frictional response of bovine articular cartilage. The main hypothesis is that CS decreases the friction coefficient of articular cartilage. Corollary hypotheses are that viscosity and osmotic pressure are not the mechanisms that mediate the reduction in the friction coefficient by CS. In Experiment 1, bovine articular cartilage samples (n=29) were tested in either phosphate buffered saline (PBS) or in PBS containing 100mg/ml of CS following 48h incubation in PBS or in PBS+100mg/ml CS (control specimens were not subjected to any incubation). In Experiment 2, samples (n=23) were tested in four different solutions: PBS, PBS+100mg/ml CS, and PBS+polyethylene glycol (PEG) (133 or 170mg/ml). In Experiment 3, samples (n=18) were tested in three solutions of CS (0, 10 and 100mg/ml). Frictional tests (cartilage-on-glass) were performed under constant stress (0.5MPa) for 3600s and the time-dependent friction coefficient was measured. Samples incubated or tested in a 100mg/ml CS solution exhibited a significantly lower equilibrium friction coefficient than the respective PBS control. PEG solutions delayed the rise in the friction coefficient relative to the PBS control, but did not reduce the equilibrium value. Testing in PBS+10mg/ml of CS did not cause any significant decrease in the friction coefficient. In conclusion, CS at a concentration of 100mg/ml significantly reduces the friction coefficient of bovine articular cartilage and this mechanism is neither mediated by viscosity nor osmolarity. These results suggest that direct injection of CS into the joint may provide beneficial tribological effects. PMID- 17084405 TI - Comparison of long-term numerical and experimental total knee replacement wear during simulated gait loading. AB - Pre-clinical experimental wear testing of total knee replacement (TKR) components is an invaluable tool for evaluating new implant designs and materials. However, wear testing can be a lengthy and expensive process, and hence parametric studies evaluating the effects of geometric, loading, or alignment perturbations may at times be cost-prohibitive. The objectives of this study were to develop an adaptive FE method capable of simulating wear of a polyethylene tibial insert and to compare predicted kinematics, weight loss due to wear, and wear depth contours to results from a force-controlled experimental knee simulator. Finite element based computational wear predictions were performed to 5 million gait cycles using both force- and displacement-controlled inputs. The displacement-controlled inputs, by accurately matching the experimental tibiofemoral motion, provided an evaluation of the simple wear theory. The force-controlled inputs provided an evaluation of the overall numerical method by simultaneously predicting both kinematics and wear. Analysis of the predicted wear convergence behavior indicated that 10 iterations, each representing 500,000 gait cycles, were required to achieve numerical accuracy. Using a wear factor estimated from the literature, the predicted kinematics, polyethylene wear contours, and weight loss were in reasonable agreement with the experimental data, particularly for the stance phase of gait. Although further development of the simplified wear theory is important, the initial predictions are encouraging for future use in design phase implant evaluation. In contrast to the experimental testing which occurred over approximately 2 months, computational wear predictions required only 2h. PMID- 17084407 TI - Verification of results to improve the quality of analytical data: a proposal. AB - After some two decades insisting on validation and normalization of methods, the reliability of the results in analytical chemistry continues to be unsatisfactory. Further, the processes involved are too slow, with the effect that frequently the most relevant results are obtained with methods not (yet) validated according to norms. It is time to rethink about how the reliability of the results could be improved. It is proposed to shift some of the control from the validation and normalization of method to the verification of the results, using verification tools built into the procedure: additional standards may enable the calculation of the yield of an extraction or chemical transformation, control the column performance or ensure that the fraction from a preseparation is correctly cut. Such verification tools provide control for every sample. If the correctness of results is verified for every sample, this confirms the adequate performance of the method and could replace a corresponding part of the validation. Examples are shown for a number of methods. PMID- 17084406 TI - Impact of pore structural parameters on column performance and resolution of reversed-phase monolithic silica columns for peptides and proteins. AB - In this work, monolithic silica columns with the C4, C8, and C18 chemistry and having various macropore diameters and two different mesopore diameters are studied to access the differences in the column efficiency under isocratic elution conditions and the resolution of selected peptide pairs under reversed phase gradient elution conditions for the separation of peptides and proteins. The columns with the pore structural characteristics that provided the most efficient separations are then employed to optimize the conditions of a gradient separation of a model mixture of peptides and proteins based on surface chemistry, gradient time, volumetric flow rate, and acetonitrile concentration. Both the mesopore and macropore diameters of the monolithic column are decisive for the column efficiency. As the diameter of the through-pores decreases, the column efficiency increases. The large set of mesopores studied with a nominal diameter of approximately 25 nm provided the most efficient column performance. The efficiency of the monolithic silica columns increase with decreasing n-alkyl chain length in the sequence of C183 (dpq)>2 (phen)>>1 (bpy). The intrinsic binding constants, obtained from UV visible spectroscopic studies, are 1.3x10(4) and 2.15 x 10(4) M(-1) for 3 and 4, respectively. Control DNA cleavage experiments using pUC19 supercoiled (SC) DNA and minor groove binder distamycin suggest major groove binding propensity for the dppz complex, while the phen and dpq complexes bind at the minor groove of DNA. Complexes 2-4 show DNA cleavage activity in dark in the presence of a reducing agent 3-mercaptopropionic acid (MPA) via a mechanistic pathway involving formation of hydroxyl radical as the reactive species. The complexes also show efficient photo-induced DNA cleavage activity on irradiation with a monochromatic UV light of 365 nm in absence of any external reagent. The cleavage efficiency follows the order: 3>4>2. The complexes exhibit significant DNA cleavage activity on irradiation with visible light of 633 nm. Control experiments show inhibition of cleavage in presence of singlet oxygen quenchers like sodium azide, histidine and enhancement of cleavage in D(2)O, suggesting formation of singlet oxygen as a reactive species in a type-II process. The photosensitizing effect of the thiomethyl group of the amino acid is evidenced from the observation of significant DNA photocleavage activity of the phen complex 2 as the phen ligand itself is not a photosensitizer. PMID- 17084457 TI - Estradiol activates mast cells via a non-genomic estrogen receptor-alpha and calcium influx. AB - BACKGROUND: Allergic airway diseases are more common in females than in males during early adulthood. A relationship between female hormones and asthma prevalence and severity has been suggested, but the cellular and molecular mechanisms are not understood. OBJECTIVE: To elucidate the mechanism(s) by which estrogens enhance the synthesis and release of mediators of acute hypersensitivity. METHODS: Two mast cell/basophil cell lines (RBL-2H3 and HMC-1) and primary cultures of bone marrow derived mast cells, all of which naturally express estrogen receptor-alpha, were examined. Cells were incubated with physiological concentrations of 17-beta-estradiol with and without IgE and allergens. Intracellular Ca(2+) concentrations and the release of beta hexosaminidase and leukotriene C(4) were quantified. RESULTS: Estradiol alone induced partial release of the preformed, granular protein beta-hexosaminidase from RBL-2H3, BMMC and HMC-1, but not from BMMC derived from estrogen receptor alpha knock-out mice. The newly synthesized LTC(4) was also released from RBL 2H3. Estradiol also enhanced IgE-induced degranulation and potentiated LTC(4) production. Intracellular Ca(2+) concentration increased prior to and in parallel with mediator release. Estrogen receptor antagonists or Ca(2+) chelation inhibited these estrogenic effects. CONCLUSION: Binding of physiological concentrations of estradiol to a membrane estrogen receptor-alpha initiates a rapid onset and progressive influx of extracellular Ca(2+), which supports the synthesis and release of allergic mediators. Estradiol also enhances IgE dependent mast cell activation, resulting in a shift of the allergen dose response. PMID- 17084460 TI - Inhibitiory effects of gold(III) ions on ribonuclease and deoxyribonuclease. AB - Inhibitory effects of gold(III) ions (Au(III)) on ribonuclease A (RNase A) and deoxyribonuclease I (DNase I) were investigated at neutral pH. RNase A was completely inhibited by 3 molar equivalents of Au(III) ions. DNase I was inhibited by 10 molar equivalents of Au(III) ions. Stoichiometric analyses suggest that Au(III) ions were coordinated to RNase A molecules. The Au(III) inhibited RNase A and DNase I were renatured to exhibit 80% and 60% of their intrinsic activity, when the bound Au(III) ions were eliminated from the nucleases by addition of thiourea, which forms a strong complex with gold ions. This suggests that RNase A and DNase I were not oxidized to lose their activity, but reversibly complexed with Au(III) ions to lose their activity. Au(III) ions were probably considered to be bound to histidine and methionine residues in the nucleases, resulting in the inhibition of their activity. CD spectra revealed that the Au(III)-induced inhibition caused a conformational change in RNase A molecules and that the addition of thiourea induced refolding of the Au(III) inhibited RNase A. PMID- 17084461 TI - Spatiotemporal pH dynamics following insertion of neural microelectrode arrays. AB - Insertion trauma is a critical issue when assessing intracortical electrophysiological and neurochemical recordings. Previous reports document a wide variety of insertion techniques with speeds ranging from 10 microm/s to 10 m/s. We hypothesize that insertion speed has an effect on tissue trauma induced by implantation of a neural probe. In order to monitor the neural interface during and after probe insertion, we have developed a silicon-substrate array with hydrous iridium oxide microelectrodes for potentiometric recording of extracellular pH (pH(e)), a measure of brain homeostasis. Microelectrode sites were sensitive to pH in the super-Nernstian range (-85.9 mV/pH unit) and selective over other analytes including ascorbic acid, Na(+), K(+), Ca(2+), and Mg(2+). Following insertion, arrays recorded either triphasic or biphasic pH(e) responses, with a greater degree of prolonged acidosis for insertions at 50 microm/s than at 0.5 mm/s or 1.0 mm/s (p<0.05). Spatiotemporal analysis of the recordings also revealed micro-scale variability in the pH(e) response along the array, even when using the same insertion technique. Implants with more intense acidosis were often associated histologically with blood along the probe tract. The potentiometric microsensor array has implications not only as a useful tool to measure extracellular pH, but also as a feedback tool for delivery of pharmacological agents to treat surgical brain trauma. PMID- 17084462 TI - Estimating health utilities and quality adjusted life years in seasonal affective disorder research. AB - BACKGROUND: Seasonal affective disorder (SAD) episodes will recur annually without effective intervention. Effectiveness of such interventions is traditionally measured with depression-specific tools (e.g., Beck Depression Inventory 2nd Edition; BDI-II). In a climate of potentially scarce resources, generic outcomes, such as Quality Adjusted Life Years (QALYs), are recommended for cost-effectiveness research. For treatments to be deemed cost-effective, they must show effectiveness relative to each other and relative to interventions across other disorders. To date, QALYs have not been used to determine effectiveness of SAD treatments. Given the recurrent nature of SAD, QALYs, which weight quality of life with time, are an ideal SAD treatment outcome. METHOD: A method to assess QALYs for SAD was developed using pilot clinical trial data. The method estimated health utilities, a measure of quality of life for a QALY, by anchoring pilot BDI-II data from the SAD clinical trial with previously derived health utilities for nonseasonal depression. RESULTS: Relative to no treatment, median QALYs gained ranged from 0.11-0.18 over 1 year, depending on the intervention assessed. DISCUSSION: Any treatment for SAD must compete with spontaneous spring remission, as illness severity attenuates in the spring. LIMITATIONS: Health utilities were estimated from the depression literature, and potential side effects from SAD treatments were not included in the estimates. The clinical trial time horizon was limited to 1-year. CONCLUSIONS: The proposed method offers researchers a tool to transform SAD efficacy data into a generic outcome for use in cost-effectiveness analysis of SAD treatments. PMID- 17084463 TI - Expressions of cyclooxygenase 1 and 2 in endotoxin-induced otitis media with effusion in the rat. AB - OBJECTIVE: Recently, a selective COX-2 inhibitor was developed and used for reducing the levels of inflammation-inducing prostaglandin (PG) whilst not inhibiting the release of protective PG by COX-1. COX-1 may be the critical isoform required for the production of PG with a homeostatic function, whereas COX-2 may be the main contributor to PG production in inflammation. The purpose of this study was to investigate COX-1 and 2 expressions in experimental endotoxin-induced OME in rats and to quantify their temporal expressions. METHODS: In a rat model, lipopolysaccharides (LPS) were inoculated into the middle ear cavity. Middle ear mucosa and temporal bone were samples at 0, 1, 3, 6, and 12h, and on days 1, 3 and 7 after instilling either LPS or sterile PBS. RT PCR, Western blotting and immunohistochemical staining were performed to determine the expressions of COX-1 and COX-2. RESULTS: COX-1 mRNA and protein were detected in normal middle ear mucosa but their levels did not change after endotoxin instillation. However, COX-2 was not identified in normal middle ear mucosa, but COX-2 mRNA was maximally increased at 6h after endotoxin instillation and COX-2 protein was maximally increased at 12h. COX-2 expression, by immunohistochemical staining, was identified only at 12h after endotoxin injection. CONCLUSIONS: In this study, the basal expressions of COX-1 and COX-2 mRNA and protein in middle ear mucosa, as well as their regulations by endotoxin were investigated. COX-1 was not induced in middle ear mucosa by endotoxin whereas COX-2 was induced within 12h of stimulation. Our findings indicate that COX-2 inhibitor administration for the relief of inflammation should be considered within 12h of the initiation of an inflammatory process. These findings may provide an understanding of the mechanisms regulating PG formation in infection of the middle ear cavity. PMID- 17084464 TI - Comparison of ELISA and immunoassays for measurement of IgG and IgM antibody to West Nile virus in human sera against virus neutralisation. AB - Two commercial assays for the detection of IgG antibody to West Nile virus (WNV), an indirect enzyme-linked immunosorbent assay (I-ELISA) and indirect fluorescent antibody test (IFAT), were evaluated against the virus neutralisation test. Excellent agreement with the virus neutralisation was obtained with both tests, i.e., 99.5% by I-ELISA and 100% by IFAT. The well-known serological cross reactivity within the family of the Flaviviridae was analysed using sera with known antibodies against dengue virus, tick borne encephalitis virus and yellow fever virus. IgM and/or IgG positive sera were examined for reactivity by WNV ELISA and WNV-IFAT. While cross-reactivity between 0 and 18.2% was recorded with IgM positive sera, there was extensive cross-reactivity of 15.7-100% with IgG positive sera. PMID- 17084465 TI - Protein serine/threonine phosphatases in neuronal plasticity and disorders of learning and memory. AB - Phosphorylation and dephosphorylation of cellular proteins by protein kinases and phosphatases represent important mechanisms for controlling major biological events. In the nervous system, protein phosphatases are contained in highly dynamic complexes localized within specialized subcellular compartments and they ensure timely dephosphorylation of multiple neuronal phosphoproteins. This modulates the responsiveness of individual synapses to neural activity and controls synaptic plasticity. These enzymes in turn play a key role in many forms of learning and memory, and their dysfunction contributes to cognitive deficits associated with aging and dementias or neurodegenerative diseases. Here, we review key modes of regulation of neuronal protein serine/threonine phosphatases and their contribution to disorders of learning and memory. PMID- 17084466 TI - The acute effects of microcystin LR on the transcription of nine glutathione S transferase genes in common carp Cyprinus carpio L. AB - The glutathione S-transferases play important roles in the detoxification of microcystin. In this experiment, nine glutathione S-transferase genes including cytosolic GSTs (rho, mu, theta, alpha and pi), mitochondrial GST (kappa) and microsomal GSTs (mGST1, mGST2 and mGST3) were cloned from common carp Cyprinus carpio. The mRNA abundance of each carp GST isoform in liver was analyzed by real time PCR. The relative changes after stimulation with microcystin LR were also analyzed: increased levels of transcription of GST alpha, rho and mGST3 isoforms were detected at 6h post stimulation; the transcription of mu, theta and mGST2 isoforms were relatively stable; and all the GST isoforms except GST kappa and rho recovered to original levels compared with controls at 72 h. It is suggested that MC-LR showed different effects on the transcription of nine carp GST isoforms. PMID- 17084467 TI - Benefit of direct ambulance to coronary care unit admission of acute myocardial infarction patients undergoing primary percutaneous intervention. AB - BACKGROUND: Early reperfusion therapy in ST-segment elevation myocardial infarction (STEMI) patients improves left ventricular (LV) function and survival. However, emergency room (ER) triage may unnecessarily delay this time-dependent treatment. We sought to determine whether direct admission of STEMI patients from the mobile intensive care units to the intensive coronary care unit (ICCU), bypassing the ER, can shorten the time intervals for primary PCI (PPCI) and improve prognosis. METHODS: All STEMI patients who underwent PPCI between Jan 2002 to Nov-2005 were included. Baseline, clinical and time interval parameters were compared between groups. Mortality rates were obtained through the population register. RESULTS: Of 533 admissions, 115 (21%) were admitted directly to the ICCU. These patients were younger (mean (+S.D.) age of 58+13 years) than patients admitted via the ER (62+/-13 years, P<0.01) and had a lower proportion of women (9% vs. 22%, P<0.01), hypertension (45% vs. 62%, P<0.01) and diabetes (15% vs. 27%, P=0.01). Directly admitted patients had a substantially shorter median pain-to-balloon time (210 vs. 247 min, P=0.02) as well as s significantly shorter door-to-balloon time (70 vs. 94 min, P<0.01), a difference that was particularly pronounced during daytime (55 min vs. 90 min, P<0.01). There were no significant differences in LV function at 24 h as assessed by echocardiography or infarct size as determined by peak creatine kinase levels. We observed a trend towards reduced 30-day (5.2% vs. 9.8%, P=0.12) and 1-year (11.1% vs. 16.1%, P=0.25) mortality in directly admitted patients. CONCLUSIONS: Directly admitted STEMI patients differ from patients admitted via the ER; Direct ICCU admission, based on a pre-hospital ECG, can substantially shorten time to treatment. PMID- 17084468 TI - A patient with tricho-rhino-phalangeal syndrome and mitral valve disease. AB - We describe the case of a 33 year old man with Tricho-Rhino-Phalangeal Syndrome (TRPS) and mitral valve disease. Tricho-Rhino-Phalangeal Syndrome is a rare multisystem disorder. The English literature has no record of any association with mitral valve disease which was a feature of our patient and his father both of whom were suffering from the syndrome. Doctors looking after patients with that condition should be alert to this possible association. PMID- 17084469 TI - Atrial fibrillation induced by carotid sinus massage. AB - A case of atrial fibrillation paroxysm induced by carotid sinus massage is reported in a patient with recurrent episodes of syncope. Despite extensive investigations, no cause for the syncopal episodes was determined. PMID- 17084470 TI - Comparison of results and complications of surgical and Amplatzer device closure of perimembranous ventricular septal defects. AB - BACKGROUND: Surgery for perimembranous ventricular septal defects (VSD) is widely accepted procedure with minimal operative mortality. Recent publications have reported the feasible, safe, and effective with the new Amplatzer VSD occluder. This study was done to compare the effectiveness, cost, and complications of both the techniques. METHODS: One hundred twenty-one consecutive patients from 2 to <18 years of age underwent VSD closure: 48 patients were treated surgically and 73 patients were treated with percutaneous Amplatzer occluder. Success rate, complications, cost, hospital stay, and home convalescent times were measured. RESULTS: The closure rate was similar in the 2 groups: 48/48 patients (100%) in the surgical group versus 71/73 patients in the Amplatzer group (97%). Procedure complications affecting management occurred in four patients of the Amplatzer group (5.5%) and four patients of surgical patients (8.3%) (p=NS). The complications that did not need treatment were observed 25/48 patients (52%) in the surgical group versus 14/73 patients (19%) in the Amplatzer group (p<0.01). Both hospital stay and home convalescent times were significantly shorter after Amplatzer closure (median hospital stay: Amplatzer three days and surgery eleven days; median convalescent time: Amplatzer two weeks and surgery six weeks). Median cost was similar for both groups. CONCLUSIONS: The closure rate was similar in the Amplatzer VSD closure and surgical closure. There were more complications in the surgical group but the majority of these was minor and did not require any change in management. Hospital stay and home convalescent times were significantly shorter after Amplatzer closure. The cost of both techniques was similar. Nevertheless, the surgeon's ability to close any VSD, regardless of anatomy, remains an important advantage of surgery. PMID- 17084471 TI - Dilemma of drug-eluting stent implantation in a patient with systemic lupus erythematosus. PMID- 17084472 TI - Evaluation of eight RNA isolation methods for transcriptional analysis in Campylobacter jejuni. AB - DNA-free RNA samples are essential to investigate gene regulation using real-time RT-PCR. This study evaluated eight RNA isolation methods in combination with TURBO DNase treatment and acid phenol extractions for their ability to produce DNA-free RNA from Campylobacter jejuni strains. PMID- 17084473 TI - A novel and in situ technique for the quantitative detection of MTBE and benzene degrading bacteria in contaminated matrices. AB - A novel and in situ technique is presented here as a better alternative to culture-dependent and PCR-based techniques for the quantitative detection of predominant bacterial species involved in the bioremediation of contaminants. It allowed rapid, specific and in situ identification of Biosep-immobilized eubacteria from MTBE- and benzene-contaminated matrices. PMID- 17084474 TI - Objective criteria to assess representativity of soil fungal community profiles. AB - Soil fungal community structures are often highly heterogeneous even among samples taken from small field plots. Sample pooling is widely used in order to overcome this heterogeneity, however, no objective criteria have yet been defined on how to determine the number of samples to be pooled for representatively profiling a field plot. In the present study PCR/RFLP and T-RFLP analysis of fungal 18S rDNA in ten soil samples obtained from a grassland plot of 400 m(2) also revealed this known heterogeneity in fungal community structures. Based on these data a three-step approach to assess representativity of fungal community profiles was established. First, soil DNA quantities needed for robust community profiling were determined. Second, profiles of single or multiple samples were theoretically averaged to test for statistically significant clustering in order to determine the minimal number of samples to be pooled to achieve representativity. Third, DNA extracts of single or multiple samples were pooled prior to profiling in order to test for statistically significant clustering. Analyses revealed robust profiles for 50 ng soil DNA but not for 5 ng. Averaged T RFLP profiles from five or more soil samples and experimental T-RFLP profiles from pools of seven or more samples formed one significant branch. Theoretical averaging and experimental pooling revealed that five to seven samples have to be pooled for robustly representing the field plot. Our results demonstrate that representativity of soil fungal community profiles can objectively be determined for a field plot with only little deviation between theoretical and experimental approaches. This three-step approach will be of assistance for designing sampling and pooling strategies for comparative analyses of soil fungal communities in ecological studies. PMID- 17084475 TI - Cell therapy and the safety of embryonic stem cell-derived grafts. AB - Recent developments in the identification, in vitro culture and differentiation of stem cells point to the unprecedented potential of these cells, or their derivatives, to cure degenerative disorders. Human embryonic stem cells (hESC) offer the particular advantage of prolonged proliferative capacity and great versatility in the lineages that can be formed in culture. Translating these advantages into clinical benefits faces many challenges, including efficient differentiation into the desired cell type(s), maintaining genetic stability during long-term culture and, finally, ensuring the absence of potentially tumorigenic hESC from the final product. It is this final safety issue that will form the focus of this review. PMID- 17084476 TI - Migration and nucleogenesis of mouse precerebellar neurons visualized by in utero electroporation of a green fluorescent protein gene. AB - Neural migration is a critical step for accurate CNS development, but the molecular mechanisms that regulate migration, settlement and nucleogenesis remain largely unknown. The precerebellar neurons (PCNs), generated in the lower rhombic lip (LRL), migrate towards their destinations: some neurons form the pontine gray nucleus (PGN) and reticulotegmental nucleus (RTN) in the ipsilateral pons, while others form the lateral reticular and external cuneate nuclei in the contralateral medulla after crossing the midline. Here, by introducing an EGFP gene into a unilateral LRL of mouse embryos by in utero electroporation, we specifically labeled and tracked the PCNs in vivo. We found that a substantial number of the labeled neurons crossed the midline and formed PGN/RTN on the contralateral side. In addition, we found that a subpopulation of the interpolar subnucleus of the spinal trigeminal nucleus, which projects the axons to the cerebellum, was one of the PCNs derived from the LRL. Furthermore, because the electroporated mice were born and grew up healthy, we could visualize the PCNs and their mossy fibers in the adult brain. Therefore, the EGFP labeling of PCNs can be applied to studying the physiology of the mossy fiber system as well as PCN development in embryos. PMID- 17084477 TI - Vitamin K1 accumulation in tobacco plants overexpressing bacterial genes involved in the biosynthesis of salicylic acid. AB - Phylloquinone (Vitamin K(1)) is an essential component of the photosynthetic electron transfer. As isochorismate is required for the biosynthesis of Vitamin K(1), isochorismate synthase (ICS) activity is expected to be present in all green plants. In bacteria salicylic acid (SA) is synthesized via a two step pathway involving ICS and isochorismate pyruvate lyase (IPL). The effect of the introduction in tobacco plants of the bacterial ICS and IPL genes on the endogenous isochorismate pathway was investigated. Transgenic tobacco plants in which IPL was targeted to the chloroplast suffered severe growth retardation and had low Vitamin K(1) content. Probably because isochorismate was channeled towards SA production, the plants were no longer able to produce normal levels of Vitamin K(1). Transgenic tobacco plants in which the bacterial ICS was present in the chloroplast showed higher Vitamin K(1) contents than wild type plants. PMID- 17084478 TI - Results of the pilot study of DIADEM: a comprehensive disease management programme for type 2 diabetes. AB - AIMS: The purpose of the study was to evaluate the feasibility and preliminary clinical effects of the DIADEM disease management programme for type 2 diabetic patients. METHODS: The study was performed at two test sites (Cardiff, UK: Aachen, Germany) including 137 and 166 patients, respectively. In 16 study centres any patients with type 2 diabetes capable of communicating by phone and able to perform blood pressure, blood glucose or urine glucose self-measurements were included. The maximum programme duration was 6 months at Cardiff and 4 months at Aachen, during which patients were assessed for glycaemic control, cardiovascular risk profile and the presence of complications of diabetes. Data were entered via the internet to a central server. RESULTS: At entry into the programme the patient group in Cardiff had significantly lower mean age (60.3+/ 9.4 years versus 64.9+/-8.7 years, p<0.001) and duration of diabetes (6.1+/-5.7 years versus 7.4+/-7.0 years, p<0.05) than in Aachen, however body mass index (31.6+/-5.2 kg/m(2) versus 29.5+/-4.9 kg/m(2), p<0.01), HbA1c (7.7+/-1.2% versus 7.1+/-1.2%, p<0.001) and systolic blood pressure (138.4+/-15.1 mmHg versus 133.5+/-11.5 mmHg, p<0.001) were significantly higher. In contrast, total cholesterol (4.7+/-1.0 mmol/l versus 5.5+/-1.1 mmol/l, p<0.001) was significantly lower in Cardiff compared to Aachen. Following entry into the programme highly significant improvements in HbA1c (Cardiff from 7.7% to 7.1%, p<0.001; Aachen from 7.2% to 6.8%, p<0.05) and total cholesterol concentrations (Cardiff: 4.66 4.46 mmol/l; Aachen: 5.33-5.15 mmol/l; both p<0.05) were observed. There were no significant changes in blood pressure at either site. CONCLUSIONS: Intensive diabetes care was delivered to DIADEM patients and relevant and significant improvements in diabetes care were achieved demonstrating that an IT-based diabetes disease management service can improve care for patients with type 2 diabetes. PMID- 17084479 TI - Glycated Collagen I (GC) impairs angiogenesis in vitro: a study using an innovative chamber for cell research. AB - Studies of cell-matrix, cell-cell interaction or angiogenesis on different matrices require simultaneous comparison of read-out parameters from differently treated companion cells. The culture conditions (cell number, temperature and volume of culture medium) in different chambers are not completely equalized using conventional methods. It has been reported that cells growing in different environmental conditions may exhibit different proliferation patterns [P. Tracqui, J.W. Liu, O. Collin, J. Clement-Lacroix, E. Planus, Global analysis of endothelial cell line proliferation patterns based on nutrient-depletion models: implications for a standardization of cell proliferation assays, Cell Proliferat. 38(June (3)) (2005) 119-135]. Herein we describe an innovative chamber, which could resolve this problem by significantly improving the standardization of experimental conditions. The chamber was manufactured from a standard cell culture well by its division with a septum into two sections. We utilized the chamber and recently developed topological analysis to examine the effects of glycated matrices on the capillary-like network formation by endothelial cells. Glycated Collagen I resulted in dose-dependent changes to all measured topological characteristics of the capillary-like network, such as the number of branching points, number of meshes and total capillary length. These differences were observed only in neighbored compartments coated with different matrices, but not in the compartments coated with the same matrix. The novel chamber brings an opportunity for better standardization of experimental conditions and simultaneous observation of different experimental groups, reducing the possible effect of any systematic error. PMID- 17084480 TI - Willingness to pay for drug abuse treatment: results from a contingent valuation study in Taiwan. AB - OBJECTIVE: In this study, we attempted to describe and justify the use of a contingent valuation (CV) method to elicit the willingness to pay (WTP) for a drug abuse treatment program by the general public in Taiwan. METHOD: In total, 1817 CV survey questionnaires were conducted through telephone interviews from randomly dialed numbers. Subjects were members of the general public aged between 20 and 65 years, with full-time jobs, and residing in the three major Taiwanese cities of Taipei, Taichung, and Kaohsiung. Respondents' WTP for drug treatment programs was elicited for two different financing mechanisms: payment through 'compulsory' payroll tax/health insurance premiums, and through 'voluntary' donations. The WTP was modeled as a function of scenarios and policies of the treatment program, respondents' socio-demographic information, and their responses to knowledge and attitudes questions. RESULTS: The general public in Taiwan was estimated to be willing to pay between NT$81.00 and NT$95.00 per month for a drug abuse treatment program, while the benefits of drug abuse treatment were estimated to range between NT$12.8 billion and NT$15.0 billion in 2004 (US$1=NT$31.9 in 2004), which was equal to around 0.15% of Taiwan's GDP for that year. The general public in Taiwan was more willing to pay for drug abuse treatment via increases in NHI premiums than via donations. Preferences for the drug abuse treatment program were also found to be sensitive to the target treated population as well as the sequence in which the WTP questions were asked. CONCLUSIONS: Results of this study provide policymakers with important evidence on the monetary value of a substance abuse treatment program, allocation of healthcare resources, and a possible financing mechanism of the treatment program, which may be justified by knowledge of the WTP of the general public. This study has also advanced the knowledge of the methodological issues with regard to CV questionnaire design, and it provides a base case for further studies on drug abuse in Taiwan. PMID- 17084481 TI - Inspiratory flow shape clustering: an automated method to monitor upper airway performance during sleep. AB - We describe an automated method for monitoring airflow dynamics in the upper airway of a sleeping subject. Its main task is to determine a set of inspiratory flow shape representatives and their relative incidence in a given respiratory airflow material. The flow shape clustering aims at reducing redundant information in the data, and thereby decreases the time needed to score overnight sleep recordings. Compared with previous computer-assisted systems, built on a pre-defined classification of prototype shapes, we require no a priori assumptions of the flow shape clusters to be discovered. The intrinsic flow shape clustering is performed with a modification of the Isodata algorithm, and the K means clustering is used as a reference in comparison studies. The operation of the method is demonstrated on clinical sleep recordings both from patients with nocturnal breathing disorders and from non-symptomatic individuals. The feasible results obtained in the practical research design suggest that application of clustering algorithms to respiratory airflow measurements could give important insights into the subtle flow shape abnormalities underlying obstructive sleep disordered breathing. PMID- 17084482 TI - Women and lung cancer: epidemiology, tumor biology, and emerging trends in clinical research. AB - Lung cancer is the leading cause of cancer-related death in both men and women. Environmental carcinogens, particularly tobacco smoke, play a dominant role in the development of lung cancer, although 10-15% of all patients diagnosed are non smokers. In addition, emerging data demonstrate sex-specific differences in lung cancer susceptibility and prognosis. This implies that the development of lung cancer is modulated by complex interactions between genetic, hormonal, behavioral, and environmental factors. A better understanding of the differences between men and women and their impact on the prevention, diagnosis, and treatment of lung cancer requires continued basic and clinical research. Recent data on the epidemiological aspects of lung cancer in women, lung tumor biology, and emerging trends in clinical research were presented at a thought leaders' roundtable hosted by the Society for Women's Health Research. The panel concluded that as the patient population in lung cancer is changing from mostly male smokers to include women and non-smokers, an urgent need exists to increase awareness and research funding to improve lung cancer care, particularly in women. To further improve survival in this disease, both clinical characteristics and tumor biology should be considered in the development of new treatment options. PMID- 17084483 TI - Distress screening in a multidisciplinary lung cancer clinic: prevalence and predictors of clinically significant distress. AB - Screening for distress in cancer patients is recommended by the National Comprehensive Cancer Network, and a Distress Thermometer has previously been developed and empirically validated for this purpose. The present study sought to determine the rates and predictors of distress in a sample of patients being seen in a multidisciplinary lung cancer clinic. Consecutive patients (N=333) were recruited from an outpatient multidisciplinary lung cancer clinic to complete the Distress Thermometer, an associated Problem Symptom List, and two questions about interest in receiving help for symptoms. Over half (61.6%) of patients reported distress at a clinically significant level, and 22.5% of patients indicated interest in receiving help with their distress and/or symptoms. Problems in the areas of family relationships, emotional functioning, lack of information about diagnosis/treatment, physical functioning, and cognitive functioning were associated with higher reports of distress. Specific symptoms of depression, anxiety, pain and fatigue were most predictive of distress. Younger age was also associated with higher levels of distress. Distress was not associated with other clinical variables, including stage of illness or medical treatment approach. Similar results were obtained when individuals who had not yet received a definitive diagnosis of lung cancer (n=134) were excluded from analyses; however, family problems and anxiety were no longer predictive of distress. Screening for distress in a multidisciplinary lung cancer clinic is feasible and a significant number of patients can be expected to meet clinical criteria for distress. Results also highlight younger age and specific physical and psychosocial symptoms as predictive of clinically significant distress. Identification of the presence and predictors of distress are the first steps toward appropriate referral and treatment of symptoms and problems that contribute to cancer patients' distress. PMID- 17084484 TI - Once-weekly epoetin beta (30,000 IU) in anemic patients with lung cancer receiving chemotherapy. AB - Anemia occurs frequently in patients with lung cancer receiving chemotherapy and has a negative impact on quality of life (QoL). Erythropoietic proteins effectively increase hemoglobin (Hb) levels, reduce transfusion requirements and improve QoL in anemic patients with a range of malignancies. This prospective, observational study evaluated epoetin beta 30,000 IU once weekly in patients with lung cancer in a real-life, clinical-practice setting. Forty patients (72.5% with NSCLC and 27.5% with SCLC) were treated with epoetin beta during any cycle of chemotherapy when Hb decreased to <12 g/dL. Hb levels were assessed at regular intervals and transfusion needs were monitored throughout the study. In total, 72.5% of patients required epoetin treatment by the second cycle of chemotherapy. Epoetin beta treatment duration ranged from 1 to >9 (median 4) weeks. Mean (+/ S.D.) baseline Hb was 10.4+/-1.2 g/dL. Epoetin beta was associated with a rapid increase in Hb levels, with a mean increase of 1.3 g/dL by week 4. Most patients (95%) remained transfusion-free throughout the study. Epoetin beta was well tolerated. This early intervention strategy with epoetin beta 30,000 IU once weekly is an effective and well-tolerated therapy for anemia in patients with lung cancer. PMID- 17084485 TI - FDG-PET maximum standardised uptake value is associated with variation in survival: analysis of 498 lung cancer patients. AB - We sought to establish the extent to which tumour uptake of [18F]-fluoro2-deoxy glucose is associated with survival in patients with primary lung cancer. From our analysis of data concerning 498 lung cancer patients, including surgical and non-surgical cases, we conclude that there is a clear association between higher tumour uptake of glucose and worse survival. PMID- 17084486 TI - Induction of cyclooxygenase-2 expression by manganese in cultured astrocytes. AB - Inflammatory and oxidative events are present in neurodegenerative disorders and appear to contribute to initiation and/or progression of the disease. Within the brain, redox-active metals, such as manganese, play an important role as components of proteins essential for neural function. However, increasing evidence implies its participation in neurodegenerative diseases involving immune modulation. Prostaglandins (PGs) are lipid mediators that participate in the regulation of physiological and pathophysiological processes, particularly during brain inflammation. In this study, we investigated whether the immune modulating action of manganese involved regulation of PGE2 production in cortical astrocytes. Within non-toxic concentrations, manganese caused an elevation in the expression of cyclooxygenase-2 (COX-2) mRNA and protein and increased PGE2 release. Manganese potentiated COX-2 expression and PGE2 generation by lipopolysaccharide/interferon-gamma-activated astrocytes. The inductive action of manganese was accompanied by generation of oxidative stress, activation of mitogen-activated protein kinases (MAPKs), AKT, and protein kinase C-alpha (PKC alpha), and increased NF-kappaB and AP-1 DNA binding activities. The generation of reactive oxygen species (ROS) was critical to manganese-induced changes in astrocytes, including MAPKs, PKC-alpha, NF-kappaB, AP-1, and COX-2 expression but not AKT. Collectively, these data indicate that manganese might cause changes in neural activity through the modulation of oxidative and inflammatory events in astrocytes. PMID- 17084487 TI - Synthesis, cytotoxicity and DNA-binding levels of ammine/propylamine platinum(II) complexes with carboxylates. AB - Seven new mixed ammine/propylamine platinum(II) complexes with carboxylates (a-g) have been synthesized and characterized by elemental analysis, conductivity, IR, UV, and (1)H NMR spectra techniques. The cytotoxicity of these complexes was tested by MTT assay. The levels of total platinum bound to DNA were measured by ICP-MS. The results indicate that the complexes (a-g) have better cytotoxicity against EJ and HL-60 than other carcinoma cell lines. The cytotoxicity increases in the sequence: g8)-catechin (procyanidin B(1)), epicatechin-(4beta-->8)-epicatechin (procyanidin B(2)), an (epi)catechin trimer, 3-O-methyl ellagic acid 4-O-beta-d-glucopyranoside, (-) epicatechin 3-O-vanillate, 3,4-methylenedioxo ellagic acid 4'-O- beta-d glucopyranoside, and 3,3'-di-O-methyl ellagic acid 4-O-beta-d-xylopyranoside. Fractionation of the raw extract by column chromatography on silicic acid yielded 10 fractions. In the hypoxanthine/xanthine oxidase antioxidant assay system, CC-9 which contained a range of polyphenols dominated by (-)-epicatechin-O-gallate proved to be the most potent antioxidant fraction (IC(50)=52 micro g/mL) in terms of ROS scavenging. In terms of XO inhibition CC-8, dominated by (epi)catechin trimer and which also contained appreciable amounts of 3'-O-methyl ellagic acid 4'-O-beta-d-xylopyranoside, as well as the catechins (+)-catechin-3-O-beta-d glucopyranoside, epicatechin-(4beta-->8)-catechin (procyanidin B(1)), and (-) epicatechin 3-O-gallate, proved to be the most potent (IC(50)=36 micro g/mL). PMID- 17084500 TI - Phospholipid fatty acid pattern and D-glucose metabolism in muscles from omega3 fatty acid-depleted rats. AB - A depletion in long-chain polyunsaturated omega3 fatty acids may affect fuel homeostasis. In such a perspective, the present study deals mainly with the in vitro fate of D-[U-(14)C]glucose in hemidiaphragms, stretched soleus and plantaris muscle pieces obtained from normal and omega3-depleted rats (second generation) and incubated in the absence or presence of insulin. When so required, the omega3-depleted rats were injected 120 min before sacrifice with either a omega3 fatty acid-rich medium-chain triglyceride:fish oil emulsion (FO) or a control medium-chain triglyceride:olive oil emulsion (OO). The content of the soleus muscle in long-chain polyunsaturated omega3 fatty acids was severely decreased in the omega3-depleted rats, and modestly albeit significantly increased after injection of FO to these animals. In stretched soleus muscles from OO-injected omega3-depleted rats, the absolute values for glycogen synthesis measured in the absence or presence of insulin were about twice higher than in normal animals. In the OO-injected omega3-depleted rats, insulin augmented the output of (14)C-labelled amino acids, whilst such was not the case in normal animals. These and other findings suggest a lower catabolism of D-glucose relative to the anabolic process of glycogen synthesis and a lower availability of endogenous amino acids in the muscles of omega3-depleted rats, as compared to those of control animals. The prior injection of FO to the omega3-depleted rats restored a normal value for the paired ratio between the output of (14)C-labelled amino acids and acidic metabolites, but further increased glycogen net synthesis. It is proposed, therefore, that the perturbation of d-glucose metabolism in muscles from omega3-depleted rats involves a multifactorial determinism, only some of the concerned factors being susceptible to rapid correction after enrichment of cell phospholipids in long-chain polyunsaturated omega3 fatty acids. PMID- 17084501 TI - Genetic analysis of polynucleotide phosphorylase structure and functions. AB - Polynucleotide phosphorylase (PNPase) is a phosphate-dependent 3' to 5' exonuclease widely diffused among bacteria and eukaryotes. The enzyme, a homotrimer, can also be found associated with the endonuclease RNase E and other proteins in a heteromultimeric complex, the RNA degradosome. PNPase negatively controls its own gene (pnp) expression by destabilizing pnp mRNA. A current model of autoregulation maintains that PNPase and a short duplex at the 5'-end of pnp mRNA are the only determinants of mRNA stability. During the cold acclimation phase autoregulation is transiently relieved and cellular pnp mRNA abundance increases significantly. Although PNPase has been extensively studied and widely employed in molecular biology for about 50 years, several aspects of structure function relationships of such a complex protein are still elusive. In this work, we performed a systematic PCR mutagenesis of discrete pnp regions and screened the mutants for diverse phenotypic traits affected by PNPase. Overall our results support previous proposals that both first and second core domains are involved in the catalysis of the phosphorolytic reaction, and that both phosphorolytic activity and RNA binding are required for autogenous regulation and growth in the cold, and give new insights on PNPase structure-function relationships by implicating the alpha-helical domain in PNPase enzymatic activity. PMID- 17084502 TI - Initiatives and outcomes of green supply chain management implementation by Chinese manufacturers. AB - This paper aims to explore the green supply chain management (GSCM) initiatives (implementation) of various manufacturing industrial sectors in China and examine the links between GSCM initiatives and performance outcomes. We conducted a survey to collect data from four typical manufacturing industrial sectors in China, namely, power generating, chemical/petroleum, electrical/electronic and automobile, and received 171 valid organizational responses for data analysis. Analysis of variance (ANOVA) was used to analyze the data. The results are consistent with our prediction that the different manufacturing industry types display different levels of GSCM implementation and outcomes. We specifically found that the electrical/electronic industry has relatively higher levels of GSCM implementation and achieves better performance outcomes than the other three manufacturer types. Implications of the results are discussed and suggestions for further research on the implementation of GSCM are offered. PMID- 17084503 TI - Properties of steel foundry electric arc furnace dust solidified/stabilized with Portland cement. AB - Electric arc furnace dust from steel production is generated in considerable amounts worldwide and needs to be treated as hazardous waste. The aim of this study was to investigate the properties of electric arc furnace dust solidified/stabilized by using Portland cement. Mortar and paste samples were prepared with varying waste-to-binder ratios between 0% and 90%. A comprehensive experimental program was designed including XRF characterization, setting time, unconfined compressive strength, and toxicity characteristics leaching procedure (TCLP), synthetic precipitation leaching procedure (SPLP), and acid neutralization capacity (ANC) tests. The results were evaluated in order to determine if the solidified /stabilized product can be disposed of at a landfill site with domestic waste or at a segregated landfill. The effect of using sand on S/S performance was also investigated. The results indicated that the solidification /stabilization process using PC helps the heavy metals to be bound in the cement matrix, but the TCLP leaching results exceeded the EPA landfilling limits. The SPLP leaching results conformed to the limits implying that the waste or S/S products can be disposed of at a segregated landfill; however the low ANC of the S/S products reveals that there may be leaching in the long-term. The sand used in the mortar samples adversely affected the S/S performance, causing higher heavy metal leaching levels, and lower pH levels in the leachate after the TCLP extraction than those measured in the leachate of the paste samples. PMID- 17084504 TI - ISO 14 001 at the farm level: analysis of five methods for evaluating the environmental impact of agricultural practices. AB - Faced with society's increasing expectations, the Common Agricultural Policy (CAP) review considers environmental management to be an ever more critical criterion in the allocation of farm subsidies. With the goal of evaluating the environmental friendliness of farm practices, France's agricultural research and extension services have built a range of agricultural/environmental diagnostic tools over recent years. The objective of the present paper is to compare the five tools most frequently used in France: IDEA, DIAGE, DIALECTE, DIALOGUE and INDIGO. All the tools have the same purpose: evaluation of the impact of farm practices on the environment via indicators and monitoring of farm management practices. When tested on a sample of large-scale farms in Picardie, the five tools sometimes produced completely different results: for a given farm, the most supposedly significant environmental impacts depend on the tool used. These results lead to differing environmental management plans and raise the question of the methods' pertinence. An analysis grid of diagnostic tools aimed at specifying their field of validity, limits and relevance was drawn up. The resulting comparative analysis enables to define each tool's domain of validity and allows to suggest lines of thought for developing more relevant tools for (i) evaluating a farm's environmental performance and (ii) helping farmers to develop a plan for improving practices within the framework of an environmental management system. PMID- 17084505 TI - Prostatic fascial anatomy and positive surgical margins in laparoscopic radical prostatectomy. PMID- 17084506 TI - The anatomic radical perineal prostatectomy: an outcomes-based evolution. AB - OBJECTIVE: Radical prostatectomy is the most effective treatment for localized prostate cancer. With increasing use of minimally invasive treatment methods, clinical outcomes are becoming important assessment tools to compare one option to another. Perineal prostatectomy is modified to incorporate contemporary surgical ideas, including preservation of cavernosal nerve bundles, sphincteric urethra at the prostatic apex, and the bladder neck. METHODS: Objective parameters and physician-reported clinical outcomes are collected prospectively on 704 consecutive patients undergoing radical perineal prostatectomy (RPP) by one surgeon. The technique described herein is the current state of evolution of RPP. The enclosed digital video is edited from two recent nerve-sparing RPPs. RESULTS: Freedom from prostate-specific antigen (PSA) detectability by stage is 94.5%, 80.0%, and 81.5% for organ-confined, specimen-confined, and margin positive disease with actual 5-yr follow-up. Margins are positive in 18% of cases. By 1, 3, 6 mo and 1 yr, 52%, 71%, 85%, and 94% of the men are free from using pads. Although >97% of nerve-spared patients have spontaneous erections, >80% can penetrate to complete intercourse. CONCLUSIONS: This method of prostatectomy is able to achieve complete cancer resection while preserving urinary and sexual function in the majority of men presenting with clinically localized prostate cancer. The simplicity and minimally invasive nature of this procedure contribute to a short recovery and low overall cost of therapy. The anatomic RPP is a cost-efficient, outcome-effective minimally invasive method of treating men with localized prostate cancer. PMID- 17084507 TI - Radical perineal prostatectomy and simultaneous extended pelvic lymph node dissection via the same incision. AB - OBJECTIVES: Assess the feasibility of extended bilateral pelvic lymph node dissection (ePLND) in radical perineal prostatectomy (RPP) via the same incision under direct vision. METHODS: In 90 consecutive patients with prostate cancer and a prostate-specific antigen level >10 ng/ml or a Gleason score >5 or more than two positive biopsies, RPP and ePLND via the same incision were performed in a prospective trial. After removing the prostate, the endopelvic fascia was opened with scissors and the bladder pushed medially. We performed an extended dissection along the obturator nerve, the external iliac vessels up to the ureter and along the internal iliac artery. Complications, number of nodes removed, and number of patients with tumour-positive nodes were recorded. Recovery of urinary continence and erectile function were assessed by a patient-reported questionnaire and the International Index of Erectile Function 5 questionnaire, respectively, administered preoperatively and at 1, 3, 6, and 12 mo. RESULTS: We removed a mean and median number of 19 and 18.7 lymph nodes, respectively. Twelve patients had lymph node metastasis. Mean operation time was 149 min, including the complete learning curves of three surgeons. Seven lymphoceles but no major complications occurred. After 1, 3, 6, and 12 mo, 32 (36%), 50 (56%), 74 (82%), and 84 (93%) patients were completely dry, using no pads. CONCLUSION: ePLND and RPP under direct vision via the same incision are feasible, efficient, and associated with a fast recovery of urinary continence and a low complication rate. Because lymphadenectomy needs no second access, the major disadvantage of RPP is resolved. PMID- 17084508 TI - Penile length alterations following penile prosthesis surgery. AB - OBJECTIVE: Determine the impact of penile prosthetic surgery on penile length. METHODS: Stretched flaccid penile length was measured in men undergoing first time penile implant surgery. Measurements were done before implantation and at 1 and 6 mo postoperatively. Patients were evaluated by the International Index of Erectile Function (IIEF) preoperatively and the IIEF and Erectile Dysfunction Inventory of Treatment Satisfaction (EDITS) at 6 mo postoperatively. Patients also provided subjective assessment of penile changes at 6 mo postoperatively. Preoperative and postoperative IIEF and EDITS scores were compared as were the patients who complained of penile length loss with those who did not. RESULTS: Of the 56 patients, 50% were diabetic and 28.5% had previous radical prostatectomy; 78% of the implants were three-piece (Alpha-1, Mentor) and 22% were two-piece (Ambicor, American Medical Service). There were no statistically significant differences in penile length after the surgery compared to preoperative measurements. Forty of 56 patients (72%) reported a decrease in penile length, 10 of 50 (19%) reported no change, and 6 of 56 (9%) had a slight increase. Subjective penile length loss was more common in patients who had undergone radical prostatectomy before prosthesis implantation (32%). No statistical difference in EF domain scores occurred between patients who complained of penile length loss and those who did not; however, men complaining of length loss had lower IIEF satisfaction domain and EDITS scores. CONCLUSION: Penile prostheses do not have a negative impact on measured stretched flaccid penile length. Treatment satisfaction scores do not depend on subjective penile length loss. PMID- 17084509 TI - Are we entering the era of truly minimally invasive destructive techniques for the treatment of urologic tumours and will these therapies stay in urologists' hands? PMID- 17084510 TI - Acetylcholine and the overactive bladder. PMID- 17084511 TI - UroVysion compared with cytology and quantitative cytology in the surveillance of non-muscle-invasive bladder cancer. AB - OBJECTIVES: The multitarget fluorescence in situ hybridization probe set Vysis UroVysion, consisting of probes for chromosomes 3, 7, and 17 and for the 9p21 band, was studied to evaluate its value in the follow-up of patients with bladder cancer. The results were compared with conventional cytology and quantitative cytology (Quanticyt). The aim of this study was to evaluate whether UroVysion is a better adjunct to urethrocystoscopy than cytology and quantitative cytology. METHODS: UroVysion, cytology, and quantitative cytology were performed on 113 voided urinary samples of 105 patients under surveillance for non-muscle-invasive bladder cancer. Before urethrocystoscopy or transurethral resection of the bladder, a voided urinary sample was obtained. Results of all tests were compared to evaluate the value of UroVysion. RESULTS: Sixty-four patients had biopsy proven urothelial cell carcinoma. Sensitivity and specificity were, respectively, 39.1% and 89.7% for UroVysion, 40.6% and 89.7% for cytology, and 42.1% and 67.9% for quantitative cytology. When the UroVysion test and cytology were combined, sensitivity increased to 53.1%, but specificity decreased to 79.5%. Detection of Ta tumours was equal for cytology and UroVysion (26.7%), detection of T1 and T2 T4 samples by UroVysion was 60% and 50%, respectively. Detection of grade 1, 2, and 3 tumours by UroVysion was 21.4%, 36.8%, and 66.7%, respectively. In four cases the UroVysion test was positive, but no abnormalities were seen at cystoscopy. CONCLUSIONS: Our data suggest that the use of UroVysion provides no improvement over cytology or quantitative cytology in the diagnosis of recurrent non-muscle-invasive bladder tumours. PMID- 17084512 TI - Failure of high-dose cyclophosphamide and etoposide combined with double-dose cisplatin and bone marrow support in patients with high-volume metastatic nonseminomatous germ-cell tumours: mature results of a randomised trial. AB - OBJECTIVES: To assess the impact on survival of high-dose chemotherapy with haematopoietic support in patients with high-volume, metastatic nonseminomatous germ cell tumours. METHODS: One hundred fifteen patients were randomised to receive either four cycles every 21 d of vinblastine (0.2 mg/kg on day 1), etoposide (100 mg/m2/d on days 1 through 5), cisplatin (40 mg/m2/d on days 1 through 5), and bleomycin (30 mg on days 1, 8, and 15) (arm A), or a slightly modified regimen followed by a high-dose chemotherapy including etoposide (350 mg/m2/d on days 1 through 5), cisplatin (40 mg/m2/d on days 1 through 5), and cyclophosphamide (1600 mg/m2/d on days 2 through 5) (arm B). RESULTS: In an intent-to-treat analysis, there were 32 (56%) and 24 (42%) complete responses in arms A and B, respectively (p=0.099). After a median follow-up of 9.7 yr, 31 and 27 patients have continuously shown no evidence of disease in arms A and B, respectively. There was no significant difference between the overall survival curves (p=0.167). According to the International Germ Cell Cancer Collaborative Group prognostic classification, the 5-yr survival rates were 88% and 82% in the intermediate group and 69% and 44% in the poor group (p=0.045) in arms A and B, respectively. CONCLUSIONS: This trial failed to demonstrate an impact on response and survival of high-dose chemotherapy with haematopoietic support in first-line treatment of patients with high-volume, metastatic nonseminomatous germ cell tumours. PMID- 17084513 TI - Minimally invasive nephron-sparing surgery (MINSS) for renal tumours. Part II: probe ablative therapy. AB - OBJECTIVES: To review the evolution and current status of probe ablative methods of minimally invasive nephron-sparing surgery (MINSS) for renal tumours. METHODS: The English language literature of the past 10 yr was reviewed by using the National Library of Medicine database and the following keywords: chemoablation, cryoablation, high-intensity focused ultrasound, kidney, laser interstitial thermotherapy, microwave thermotherapy, nephron-sparing surgery, radiofrequency ablation, radiosurgery, renal, and tumour. Over 300 papers were identified, 50 of which were selected for this review on the basis of their contribution in advancing the field with regards to (1) evolution of concepts, (2) development and refinement of techniques, and (3) intermediate- and long-term clinical outcomes. RESULTS: Open partial nephrectomy is the reference standard for nephron sparing surgery against which all MINSS techniques should be measured. Although the initial outcomes of cryoablation and radiofrequency ablation (RFA) are encouraging, long-term studies are necessary to confirm lasting efficacy. The optimal modality for tumour targeting, monitoring therapy, and follow-up remains to be determined. These ablative techniques should be reserved for carefully selected patients, the data should be prospectively accrued, and the long-term cancer cure rates should be compared with the reference standard. CONCLUSIONS: Promising long-term data are available for cryoablation. RFA is still considered developmental, and instances of incomplete cell kill, despite nonenhancement, are concerning. Other modalities are still experimental. PMID- 17084515 TI - Intermediate-term results of retroperitoneoscopy-assisted cryotherapy for small renal tumours using multiple ultrathin cryoprobes. AB - OBJECTIVES: To evaluate outcome after retroperitoneoscopic cryotherapy for small renal tumours. METHODS: Fourteen patients underwent cryoablation performed with six ultrathin 1.5-mm cryoprobes. Retroperitoneoscopic access was used for any tumour location. A double freeze-thaw cycle was performed under simultaneous retroperitoneoscopic visual control and real-time ultrasound monitoring. RESULTS: Mean tumour size was 2.8 cm (range: 2.0-4.0), mean patient age was 68 yr (range: 49-83), and six left and eight right kidneys were treated. The mean operative time was 167 min (range: 120-200); mean blood loss was 93 ml (range: 0-300). Cryosurgery was successfully performed in all 14 patients, with 13 patients undergoing assisted retroperitoneoscopy and one patient, after previous open nephropexy, undergoing an open approach. Intraoperative biopsy specimens revealed renal cell cancer (RCC) in 10 (71%) patients. The only intraoperative complication was bleeding after removal of the cryoprobes in four (29%) patients, necessitating one intracorporeal stitch in each. Two (14%) of the first four patients presented postoperatively with superficial skin frostbite, which healed with conservative treatment. During mean follow-up of 21 mo (range: 2-42), 2 (14%) patients died from unrelated disease, and 12 patients remained without evidence of local recurrence. One patient with previous contralateral nephrectomy for RCC developed retroperitoneal lymph node metastasis on that side without recurrence in the cryoablated kidney. CONCLUSIONS: Retroperitoneoscopic cryotherapy using multiple ultrathin cryoprobes is safe and effective with encouraging oncologic results on intermediate-term follow-up. Prospective clinical trials and long-term oncologic data are needed to define its definitive role in treatment of renal tumours. PMID- 17084516 TI - Are abstracts presented at the EAU meeting followed by publication in peer reviewed journals? A critical analysis. AB - PURPOSE: Our goal was to assess the rate and time-course of peer-reviewed publication of abstracts presented at the European Association of Urology (EAU) Annual Meeting and to identify factors predictive of publication. METHODS: All abstracts accepted for presentation at the 2000 and 2001 EAU annual meetings were identified from the related published supplements in European Urology. The subsequent publication rate was estimated for the corresponding studies based on a scan of Medline covering a 5-year period following the meetings. We examined whether the following factors were associated with publication rate: research type, study subject, and country of origin. The analysis was performed using logistic regression of the dichotomous variable of publication versus non publication and the candidate factors. RESULTS: Overall, 47.3% of 1406 abstracts presented at the EAU meetings were followed by publication in peer-reviewed journals. Pre-clinical research studies were more likely to be published than clinical studies (53.3% vs 45%, p<0.05). Prospective series were more likely to be published than retrospective ones (46.5% vs 32.2%, p<0.05). Studies presented at the meetings were mostly from Europe (74.2%). Mean time to publication was 8.6 months, and in most cases, the reports were published in The Journal of Urology and European Urology. The mean IF of journals where papers were published was 1.95. CONCLUSIONS: Almost half of the abstracts presented at the EAU are ultimately published in peer-reviewed journals, usually within 2 years after presentation. The publication rate differs significantly according to country of origin, study subject, and research type. PMID- 17084517 TI - A sealed laparoscopic nephroureterectomy: a new technique. AB - OBJECTIVES: To describe a purely laparoscopic nephroureterectomy approach that avoids the disadvantages of transurethral bladder cuff excision and open/laparoscopic distal ureterectomy using the EndoGIA. METHODS: A standard transperitoneal laparoscopic nephrectomy is carried out through three or four ports in the flank. The ureter is dissected caudally into the pelvis. Two additional (5- and 10-mm) trocars are placed in the ipsilateral lower abdomen. Caudal ureteral dissection continues until the detrusor muscle fibers at the ureterovesical junction are identified. A 1-cm area of bladder adventitia around the ureterovesical junction is cleared. The ureter is retracted upward and laterally, tenting up the bladder wall. The bladder cuff is excised using a 10-mm LigaSure Atlas and detached from the bladder. A 6-cm lower-quadrant incision is used to remove the specimen in an Endocatch bag. An indwelling 16F Foley catheter is then placed. RESULTS: Thirteen adult patients with suspected upper-tract transitional cell carcinoma underwent this surgical technique (operative time: 170-270 min): none had local recurrence, and two had recurrence remote from the bladder cuff scar (follow-up: 1-23 months). CONCLUSIONS: The described procedure adheres strictly to oncologic principles (removal of the affected renal unit without opening the urinary tract), and circumvents the need for transurethral/intraureteral instrumentation and patient repositioning. PMID- 17084518 TI - Therapeutic effectiveness and patient satisfaction after 6 months of treatment with tadalafil, sildenafil, and vardenafil: results from the erectile dysfunction observational study (EDOS). AB - OBJECTIVE: This observational study was conducted across Europe to assess health outcomes in men with erectile dysfunction (ED) who took tadalafil, sildenafil citrate (sildenafil), or vardenafil HCl (vardenafil) for 6 mo. METHODS: Therapy effectiveness and patient satisfaction were evaluated using established and new questions on erectile function. Behavioural, psychological, and relationship outcomes were assessed using the short form of the Psychological and Interpersonal Relationship Scales (SF-PAIRS). RESULTS: In nine European countries at 904 sites, 8047 patients were enrolled and 94% (7560) selected either tadalafil (5315), sildenafil (1252), or vardenafil (993) for treatment at baseline. Of the 7560, 3998 (52.9%) took the same drug for 6 mo. Baseline characteristics across the three treatment groups were comparable: mean age approximately 56 yr, moderate or severe ED, and mean International Index of Erectile Function-Erectile Function domain score about 13. Tadalafil, sildenafil, and vardenafil were therapeutically effective and improved patient satisfaction in the 40-58% of men who completed 6 mo of a single therapy. Patients taking tadalafil consistently had numerically higher levels of therapeutic effectiveness and satisfaction compared with patients who took sildenafil or vardenafil. The three cohorts had statistically significant changes from baseline in response to SF-PAIRS and there were significant differences, in favour of tadalafil, among cohorts in the Time Concerns domain. CONCLUSION: In a large observational study that mimics a routine clinical setting, most patients selected an inhibitor of phosphodiesterase 5 to treat ED, which resulted in a high level of therapeutic effectiveness and patient satisfaction. PMID- 17084519 TI - Acetylcholine and molecular components of its synthesis and release machinery in the urothelium. AB - OBJECTIVES: Previous studies provided indirect evidence for urothelial synthesis and release of acetylcholine (ACh). We aimed to determine directly the ACh content in the urothelium and to characterize the molecular components of its synthesis and release machinery. METHODS: The study was performed on mouse bladder and abraded urothelium, and human mucosal bladder biopsies. ACh content was measured by high-performance liquid chromatography-electrochemical. Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry served to investigate expression of ACh-synthesizing enzymes-choline acetyltransferase (ChAT) and carnitine acetyltransferase (CarAT)-vesicular ACh transporter (VAChT), and polyspecific organic cation transporters (OCTs; isoforms 1-3). Transfected cells served to investigate whether the anticholinergic drug trospium chloride interferes with ACh-transporting OCTs. RESULTS: ACh is present in the urothelium in a nanomolar range per gram of wet weight. RT-PCR data support the presence of CarAT but not ChAT. VAChT, used by neurons to shuffle ACh into synaptic vesicles, is detected in subepithelial cholinergic nerve fibres, but not by RT-PCR or immunohistochemistry in the urothelium. OCT1 and OCT3 are expressed by the urothelium. The quarternary ammonium base trospium chloride inhibits cation transport by OCTs with a potency rank order of OCT2 (IC(50)=0.67+/ 0.42micromol/l)>OCT1 (IC(50)=6.2+/-2.1micromol/l)>OCT3 (IC(50)=871+/ 177micromol/l). CONCLUSIONS: This study demonstrates a urothelial non-neuronal cholinergic system that differs widely from that of neurons with respect to molecular components of the ACh synthesis and release machinery. Consequently, these two systems might be differentially targeted by pharmacologic approaches. PMID- 17084520 TI - Robotic-assisted laparoscopic prostatectomy: functional and pathologic outcomes with interfascial nerve preservation. AB - INTRODUCTION: Robotic-assisted laparoscopic radical prostatectomy (RLRP) is increasingly becoming an alternative to open and laparoscopic radical prostatectomy in the treatment of localized prostate cancer. RLRP has been associated with low morbidity, short convalescence and comparable oncologic and functional outcomes. We report our initial experience of 300 consecutive cases with selective use of interfascial nerve preservation (IFNP). METHODS: Between February 2003 and September 2005, 300 consecutive men underwent RLRP at our institution. Patients were followed prospectively with validated questionnaires. RESULTS: Mean operative time was 282 minutes with an estimated blood loss of 273 ml. The intra-operative complication rate was 2.3% with no mortality. Return to baseline (RTB) urinary function and subjective continence at 12 months were 71% and 90.2%, respectively. RTB sexual function and subjective potency at 12 months were 53% and 80.4%, respectively. Overall, the positive surgical margin (PSM) rate was 20.9%: 15.1% for pT2 and 52.1% for pT3 disease and 93.1% had an undetectable PSA (<0.1 ng/mL) with a mean follow-up of 17.3 months. Fifty-four percent of PSMs occured in a poster-lateral (PL) location. Retrospectively, IFNP was performed in 86.5% and 62.5% of pT2 and pT3 PSMs, respectively. Pathologic-T3 PSMs were found to occur significantly more often in a PL location when ipsilateral IFNP was performed when compared to non-IFNP (73% vs 33%, p=0.05). CONCLUSIONS: IFNP appears to result in favorable return of potency, however, postero-lateral PSMs appear to occur more frequently with this technique. Proper patient selection for robotic surgery and nerve-preservation appears to be critical in order to reduce PSM and optimize the oncologic efficacy of this technology. PMID- 17084521 TI - Onconase induces caspase-independent cell death in chemoresistant neuroblastoma cells. AB - The efficacy of Onconase on the growth of a panel of chemosensitive and chemoresistant neuroblastoma cell lines was investigated. Onconase decreased cell viability of chemosensitive (IMR-32, UKF-NB-3) and chemoresistant neuroblastoma cell lines characterised by high expression of P-glycoprotein (P-gp) (UKF-NB 3(r)DOX(20)) or by high P-gp expression in combination with mutated p53 (UKF-NB 3(r)VCR(10), Be(2)-C), in a similar manner. Moreover, Onconase caused cell cycle block in G1 phase and induced caspase-independent cell death. Transmission electron microscope investigations suggested that Onconase-induced autophagy contributes to Onconase-induced cell death. Antitumour activity of Onconase against naive and drug-resistant neuroblastoma xenografts was confirmed in animals. PMID- 17084522 TI - Decreased aquaporin expression leads to increased resistance to apoptosis in hepatocellular carcinoma. AB - Cells undergoing apoptosis are characterized by decreased cell size due to changes in intracellular ion concentration and rapid, aquaporin (AQP)-dependent water movement out of the cell, events required for the activation of pro apoptotic enzymes. The current study demonstrates AQP 8 and 9 expression is significantly decreased in hepatocellular carcinoma (HCC) versus normal liver. Isolation of hepatic tumor cells (H4IIE) and hepatocytes confirmed a lack of water movement across the H4IIE cell membrane via AQPs and identified an inherent resistance of H4IIE cells to apoptotic stimuli. In contrast, hepatocytes rapidly responded to osmotic challenge through AQP-dependent water movement and underwent cell death following apoptotic stimulation. PMID- 17084523 TI - Recovery of metals from Cuban nickel tailings by leaching with organic acids followed by precipitation and magnetic separation. AB - The percolation leaching of the Cuban nickel tailings containing 0.34% Ni, 0.08% Co and 44.2% Fe was investigated by using tartaric and oxalic acids at different concentrations. About 70% Ni, 80% Co and 30% Fe were extracted after 5 days of leaching with the mixture of 0.15 mol/L tartaric acid and 0.05 mol/L oxalic acid at ambient temperature and normal pressure. Nickel and cobalt extraction of 80% as well as iron extraction of 50% were achieved from the pregnant solution by means of precipitation at 80 degrees C for 2h. The precipitation at ambient temperature led to a similar result after 16 days. Cobalt, nickel and iron oxalates were found in the precipitate by using the X-ray diffraction method. The regeneration of acids during the precipitation step made possible the reuse of the raffinate at the leaching step. Heating of the precipitate at 200 degrees C increased the metal concentration to 1.22% Ni and 0.33% Co, which can be fed in the existing nickel plant in Moa, Cuba. The magnetic processing of the leaching residues led to a non-magnetic product containing less than 20% Fe and a magnetic product containing more than 50% Fe. PMID- 17084524 TI - Using UV pretreatment to enhance biofiltration of mixtures of aromatic VOCs. AB - Mixtures of airborne toluene and o-xylene, two relatively recalcitrant volatile organic compounds (VOCs), were treated effectively using integrated UV biofiltration. The set-up consisted of a biofilter receiving UV-pretreated stream and a reference biofilter receiving no pretreatment. Experimental conditions included UV fluences of 6 and 12 mJcm(-2) as well as air flow rates of 6.3 and 9.4 Lmin(-1), corresponding to biofilter empty bed retention times (EBRTs) of 45 and 30s, respectively. The inlet concentration of organics (toluene and o-xylene) ranged between 70 and 650 mg(carbon)m(-3). The UV-biofilter consistently provided removal efficiencies of greater than 95% over the range of toluene and o-xylene inlet concentrations. Also, the coupled UV-biofiltration system provided up to 60% additional contaminant removal compared to the sum of that offered by UV and reference biofilter, demonstrating the synergistic effect of UV on biofilter performance. The UV photooxidation partially oxidized a fraction of toluene and o xylene into water soluble and more biodegradable intermediates, such as acetaldehyde and formaldehyde, which were readily removed in the downstream biofilter. These intermediates along with up to 20ppmv ozone, formed through the photolysis of oxygen by 185 nm UV, contributed to the enhanced degradation of parent VOCs in the biofilter as well as the absence of any inhibitory effects of the VOCs on one another. Also, the presence of ozone helped control the growth of excess biofilm in the UV-coupled biofilter. While the standalone biofilter showed significant pressure drop increase (of up to 14 mm H(2)Om(-1) of the bed) over the course of experiment, the UV-coupled biofilter maintained a relatively low pressure drop of less than 3 mmH(2)Om(-1) of the bed. PMID- 17084525 TI - Study of imidaclopride removal from aqueous solution by adsorption onto granular activated carbon using an on-line spectrophotometric analysis system. AB - The removal of imidaclopride as a pesticide by granular activated carbon (GAC) and its adsorption kinetics were studied at different pH values and temperatures. In all experiments, the amount of GAC and initial concentration of imidaclopride were 2g and 25 ppm, respectively. The adsorption process was followed by an on line spectrophotometric analysis system, which consisted of UV-spectrophotometer, a designed absorption cell, peristaltic pump and special glassy reactor. The effect of pH and temperature on adsorption was studied over 90 min adsorption periods. The obtained data were treated according to various kinetic models. The results showed that second order model was the most suitable one on the overall. The our results also showed that the adsorption rate constants for first order, second order and intraparticle diffusion models followed decreasing order: pH=7>4>10>1, T=25>35>45>55 degrees C. PMID- 17084526 TI - Application of response surface methodology for the biosorption of copper using Rhizopus arrhizus. AB - Response surface methodology was used to study the cumulative effect of the various parameters namely, initial copper ion concentration, pH, temperature, biomass loading and to optimize the process conditions for the maximum removal of copper. For obtaining the mutual interaction between the variables and optimizing these variables, a 2(4) full factorial central composite design using response surface methodology was employed. The analysis of variance (ANOVA) of the quadratic model demonstrates that the model was highly significant. The model was statistically tested and verified by experimentation. A maximum copper removal of 98% was obtained using the biosorption kinetics of copper under optimum conditions. PMID- 17084527 TI - Effect of temperature on the release of hexadecane from soil by thermal treatment. AB - A natural organic soil (2.5% of total organic carbon) was artificially contaminated with hexadecane, and thermally treated under an inert medium up to different final temperatures (150-800 degrees C) for 30 min to simulate ex situ thermal process conditions. The experiments were conducted using a complete organic soil, instead of the clays or isolated soil fractions that are commonly used. Neat and contaminated samples were separately heated to understand the impact of the soil itself and the contaminant in the release of volatiles. The soil quality as well as the quality and amount of volatile compounds generated during the process were monitored. More than 80-88% of the initial hexadecane content in the soil matrix was recovered in liquids traps after the thermal treatment, therefore the contaminant could be recovered for further recycling. The high amount of hexadecane collected without suffering chemical transformations indicated that the main mechanism for the hexadecane removal was evaporation. The analysis of the light gases released from contaminated samples indicated negligible or null hexadecane pyrolysis reaction rates, confirming that the evaporation/desorption of the contaminant are the processes that governed the removal of the contaminant from the soil. For the soil tested, of a relatively low surface area, good removal efficiencies (higher than 99.9%) were detected at about 300 degrees C, being higher temperatures not necessary to significantly improve the contamination removal. PMID- 17084528 TI - Peroxisome proliferator-activated receptor-gamma2 Pro12Ala polymorphism is associated with reduced risk for ischemic stroke with type 2 diabetes. AB - Peroxisome proliferator-activated receptor (PPAR)-gamma2 has important effects to insulin sensitivity, atherosclerosis, inflammation and endothelial cell function. Through these effects, PPAR-gamma2 might be involved with the ischemic stroke in type 2 diabetes. To determine the role of PPAR-gamma2 in genetic susceptibility to ischemic stroke in type 2 diabetes, we genotyped 302 ischemic stroke patients, 283 healthy controls and 141 type 2 diabetic patients without ischemic stroke (diabetes duration >10 years) for PPAR-gamma2 Pro12Ala polymorphism by polymerase chain reaction-restriction fragment length polymorphism methods. PPAR-gamma2 Pro/Ala genotype were lower in ischemic stroke patients than those observed in the control group (4.0% vs. 9.9%, OR=0.38, P=0.0046), and it were associated with the incidence of ischemic stroke in the multivariate analysis (OR=0.43, P=0.025). Genotypic analysis revealed that ischemic stroke patients with type 2 diabetes displayed a great lower prevalence of the Pro/Ala genotype (2.3%) than controls (9.9%) (OR=0.21, P=0.0047). And Pro/Ala genotype of type 2 diabetes patients with ischemic stroke were lower than type 2 diabetes patients without ischemic stroke (2.3% vs. 8.5%, OR=0.25, P=0.0321), however the significant association with ischemic stroke was not detected in the multivariate analysis (OR=0.27, P=0.051). These results suggest that the Pro/Ala genotype of PPAR-gamma2 Pro12Ala polymorphism may be associated with reduced risk for ischemic stroke, and the possibility that it might have a protective effect for ischemic stroke with type 2 diabetes. PMID- 17084529 TI - Early dysregulation of cripto-1 and immunomodulatory genes in the cerebral cortex in a macaque model of neuroAIDS. AB - Human immunodeficiency virus type 1 (HIV-1) and related primate lentiviruses are known to enter the central nervous system (CNS) during the primary phase of infection. Neuroinvasion by simian immunodeficiency virus and simian human immunodeficiency virus (SHIV) is characterized by transient meningitis and astrocytosis. In this report, we used targeted cytokine cDNA arrays to analyze cortical brain tissue from four pig-tailed macaques inoculated for 2 weeks with pathogenic SHIV(50OLNV) and a normal age-matched pig-tailed macaque. Our results revealed that eight genes were significantly upregulated in all four macaques. These included: leukocyte interferon inducible peptide, corticotrophin releasing factor receptor 1, interleukin 6, CDW40 antigen, cysteine-rich fibroblast growth factor, neurotrophin 3, ciliary neurotrophin factor receptor and cripto-1. The upregulation of three of these genes was confirmed by reverse transcriptase PCR (RT-PCR). Since cripto-1 had not been previously identified within specific cell types within the primate central nervous system, we performed immunohistochemical studies, which revealed the presence of cripto-1 in neurons. RT-PCR studies demonstrated that cripto-1 mRNA was widely expressed in the CNS. These results indicate that immunomodulatory genes are upregulated during the primary phase of infection of the central nervous system. Cripto-1, which acts as a survival factor in tumor cells and may be neuroprotective, is expressed in neurons within the CNS and is upregulated during viral invasion. PMID- 17084530 TI - GABA-induced intracellular Ca2+ elevation in the embryonic chick brainstem slice. AB - We examined the intracellular Ca2+ ([Ca2+]i) elevation evoked by GABA in an 8-day embryonic chick brainstem slice using a Ca imaging technique with Ca green-1 AM. When small quantities of GABA were pressure-ejected on the surface of the slice, the [Ca2+]i elevation was clearly detected. The GABA-induced [Ca2+]i elevation was eliminated in a Ca2+-free solution, whereas the previously reported GABA induced light-scattering change was independent of extracellular Ca2+. Although, micro-application of glycine or glutamate also induced [Ca2+]i elevation, these changes were smaller than that by GABA. These results suggest that the GABA induced [Ca2+]i elevation is due to Ca2+ entry resulting from membrane depolarization and may play an important role in the development of the central nervous system (CNS). PMID- 17084531 TI - An artificial three-dimensional matrix promotes ramification in the microglial cell-line, BV-2. AB - Three-dimensional (3D) cell culture yields strikingly different cell phenotypes compared to two-dimensional (2D) cell culture. Since microglia, monocyte derived immune cells in the brain, exist in a variety of cell shapes ranging from amoeboid to ramified, we evaluated the impact of 2D versus 3D culture conditions on cell shape. The microglial cell-line, BV-2, was either cultured on poly-D lysine coated dishes (2D culture conditions) or in a BD Pura Matrix Peptide Hydrogel (3D culture conditions) in the absence or presence of the extracellular matrix proteins, fibronectin and collagen type I, respectively. We identified five distinct morphological phenotypes (amoeboid, bipolar, tripolar, multipolar, ramified) and compared the frequency distribution of these phenotypes under different culture conditions using a chi(2) test. Culture of BV-2 cells in an inert 3D matrix shifted the frequency distribution from an amoeboid dominated population, which is typical for BV-2 cells cultured under conventional 2D conditions, to a population dominated by multipolar phenotypes. Fibronectin or collagen type I significantly suppressed matrix-induced ramification. These cell culture experiments illustrate the dependency of cell shape on spatial distribution of potential adhesion sites. PMID- 17084532 TI - Perinatal priming of calves born to Schistosoma mattheei-infected dams. AB - The objective of this study was to elucidate whether calves born to infected dams had been primed against Schistosoma mattheei antigens. Infection-confirmed, pregnant cows were randomly selected for monitoring their offspring. Pre colostral serum was collected from the neonates for the detection of specific antibodies at birth, as they indicate a transplacental transfer of schistosome specific antibodies and antigen. At the age of approximately 2 months, peripheral blood mononuclear cells (PBMC) of calves were analysed for specific memory by antigen-specific stimulation in vitro. Twenty-six of the 30 calves demonstrated S. mattheei-specific proliferation. All 12 seropositive-born, as well as 14 of the 18 seronegative-born (before colostrum uptake) calves displayed mattheei specific proliferation. The results indicate that the calves were primed against S. mattheei and might explain why seropositive-born calves from infected dams are better protected against S. mattheei, and query the impermeability of the damaged ruminant placenta with consequences for antigen transfer. PMID- 17084533 TI - Mosquito-net burns and the prevention hexagon. PMID- 17084534 TI - Molecular mechanisms of resistance and toxicity associated with platinating agents. AB - Platinating agents, including cisplatin, carboplatin, and oxaliplatin, have been used clinically for nearly 30years as part of the treatment of many types of cancers, including head and neck, testicular, ovarian, cervical, lung, colorectal and relapsed lymphoma. The cytotoxic lesion of platinating agents is thought to be the platinum intrastrand crosslink that forms on DNA, although treatment activates a number of signal transduction pathways. Treatment with these agents is characterized by resistance, both acquired and intrinsic. This resistance can be caused by a number of cellular adaptations, including reduced uptake, inactivation by glutathione and other anti-oxidants, and increased levels of DNA repair or DNA tolerance. Here we investigate the pathways that treatment with platinating agents activate, the mechanisms of resistance, potential candidate genes involved in the development of resistance, and associated clinical toxicities. Although the purpose of this review is to provide an overview of cisplatin, carboplatin, and oxaliplatin, we have focused primarily on preclinical data that has clinical relevance generated over the past five years. PMID- 17084536 TI - Organization of telencephalotectal projections in pigeons: Impact for lateralized top-down control. AB - Birds display hemispheric specific modes of visual processing with a dominance of the right eye/left hemisphere for detailed visual object analysis. In pigeons, this behavioral lateralization is accompanied by morphological left-right differences in the ascending tectofugal pathway. This system is also asymmetrically modulated by descending telencephalotectal input whereby the left forebrain displays a much more pronounced physiological control over ipsilateral left and contralateral right visual thalamic processes. In the present study we aimed to answer the question if this top-down asymmetry that up to now had been demonstrated in single cell recording studies is due to anatomical asymmetries in the size of the fiber systems descending from the telencephalon to the tectum. We approached this question by means of a quantitative retrograde tracing study. Cholera toxin subunit B (CtB) was injected unilaterally into either the left or right optic tectum of adult pigeons. After immunohistochemical detection of CtB positive cells, the number of ipsi- and contralaterally projecting neurons was estimated. Retrogradely labeled cells were located within the arcopallium, the hyperpallium apicale (HA) and the temporo-parieto-occipital area (TPO). Descending projections from HA, arcopallium, and TPO were mainly or exclusively ipsilateral with the contralateral projection being extremely small. Moreover, there was no difference between left and right hemispheric projections. These anatomical data sharply contrast with behavioral and electrophysiological ones which reveal an asymmetric and bilateral top down control. Therefore, contralateral and lateralized forebrain influences onto tectofugal processing are possibly not the direct result of asymmetrical descending axon numbers. Those influences emerge by a lateralized intra- and/or interhemispheric integration of ascending and descending input onto the rotundus. PMID- 17084535 TI - Chemotherapy-evoked painful peripheral neuropathy: analgesic effects of gabapentin and effects on expression of the alpha-2-delta type-1 calcium channel subunit. AB - Chemotherapeutics in the taxane and vinca-alkaloid classes sometimes produce a painful peripheral neuropathy for which there is no validated treatment. Experiments with rat models of paclitaxel- and vincristine-evoked pain suggest that these conditions may not respond to all of the analgesics that have efficacy in other models of painful peripheral neuropathy. We tested gabapentin as a potential analgesic for paclitaxel- and vincristine-evoked pain. We used a repeated dosing paradigm because there are precedents showing that repeated drug exposure may be necessary to demonstrate analgesia in neuropathic pain models. Gabapentin is believed to work via binding to voltage-gated calcium channels that contain the alpha-2-delta type-1 (alpha(2)delta-1) subunit, and the expression of this subunit is known to be increased in some painful peripheral neuropathy models. Thus we also examined whether the paclitaxel-evoked pain syndrome was accompanied by an alpha(2)delta-1 increase, and whether gabapentin had any effect on subunit expression. We found that the paclitaxel- and vincristine-evoked mechano-allodynia and mechano-hyperalgesia were significantly reduced by gabapentin, but only with repeated dosing. Paclitaxel-evoked painful peripheral neuropathy was associated with an increased expression of the alpha(2)delta-1 subunit in the spinal dorsal horn, but not in the dorsal root ganglia. The spinal cord increase was normalized by repeated gabapentin injections. Together, these findings suggest that repeated dosing with gabapentin may be beneficial in patients with chemotherapy-evoked painful peripheral neuropathy and that gabapentin's mechanisms of action may include normalization of the nerve injury evoked increase in calcium channel alpha(2)delta-1 subunit expression. PMID- 17084537 TI - Delayed riluzole treatment is able to rescue injured rat spinal motoneurons. AB - The effect of delayed 2-amino-6-trifluoromethoxy-benzothiazole (riluzole) treatment on injured motoneurons was studied. The L4 ventral root of adult rats was avulsed and reimplanted into the spinal cord. Immediately after the operation or with a delay of 5, 10, 14 or 16 days animals were treated with riluzole (n=5 in each group) while another four animals remained untreated. Three months after the operation the fluorescent dye Fast Blue was applied to the proximal end of the cut ventral ramus of the L4 spinal nerve to retrogradely label reinnervating neurons. Three days later the spinal cords were processed for counting the retrogradely labeled cells and choline acetyltransferase immunohistochemistry was performed to reveal the cholinergic cells in the spinal cords. In untreated animals there were 20.4+/-1.6 (+/-S.E.M.) retrogradely labeled neurons while in animals treated with riluzole immediately or 5 and 10 days after ventral root avulsion the number of labeled motoneurons ranged between 763+/-36 and 815+/-50 (S.E.M.). Riluzole treatment starting at 14 and 16 days after injury resulted in significantly lower number of reinnervating motoneurons (67+/-4 and 52+/-3 S.E.M., respectively). Thus, riluzole dramatically enhanced the survival and reinnervating capacity of injured motoneurons not only when treatment started immediately after injury but also in cases when riluzole treatment was delayed for up to 10 days. These results suggest that motoneurons destined to die after ventral root avulsion are programmed to survive for some time after injury and riluzole is able to rescue them during this period of time. PMID- 17084539 TI - Functional outcome and survival after radiotherapy of metastatic spinal cord compression in patients with cancer of unknown primary. AB - PURPOSE: Patients with cancer of unknown primary (CUP) account for about 10% of patients with metastatic spinal cord compression (MSCC). This study aims to define the appropriate radiation regimen for these patients. METHODS AND MATERIALS: Data of 143 CUP patients irradiated for MSCC were retrospectively evaluated. Short-course radiotherapy (RT) (1x8 Gy, 5x4 Gy, n = 68) and long course RT (10x3 Gy, 15x2.5 Gy, 20x2 Gy, n = 75) plus 8 further potential prognostic factors (age, gender, performance status, visceral metastases, other bone metastases, number of involved vertebrae, ambulatory status, time of developing motor deficits before RT) were compared for functional outcome and survival. RESULTS: Improvement of motor function occurred in 10% of patients, no further progression of motor deficits in 57%, and deterioration in 33%. On multivariate analysis, functional outcome was positively associated with slower development of motor deficits (p < 0.001), absence of visceral metastases (p = 0.008) and other bone metastases (p = 0.027), and ambulatory status (p = 0.054), not with the radiation regimen (p = 0.74). Recurrence of MSCC in the irradiated region occurred in 7 patients after median 6 months. Median survival was 4 months. On multivariate analysis, better survival was significantly associated with absence of visceral metastases (p < 0.001), absence of other bone metastases (p = 0.005), ambulatory status (p = 0.001), and slower development of motor deficits (p = 0.030). CONCLUSIONS: For MSCC treatment in patients with CUP, no significant difference was observed between short-course and long-course RT regarding functional outcome and survival. Short-course RT appears preferable, at least for patients with a poor predicted survival, as it is more patient convenient and more cost-effective. PMID- 17084538 TI - L-carnitine protects neurons from 1-methyl-4-phenylpyridinium-induced neuronal apoptosis in rat forebrain culture. AB - 1-Methyl-4-phenylpyridinium ion (MPP+), an inhibitor of mitochondrial complex I, has been widely used as a neurotoxin because it elicits a severe Parkinson's disease-like syndrome with an elevation of intracellular reactive oxygen species (ROS) and apoptosis. L-carnitine plays an integral role in attenuating the brain injury associated with mitochondrial neurodegenerative disorders. The present study investigates the effects of L-carnitine against the toxicity of MPP+ in rat forebrain primary cultures. Cells in culture were treated for 24 h with 100, 250, 500 and 1000 microM MPP+ alone or co-incubated with L-carnitine. MPP+ produced a dose-related increase in DNA fragmentation as measured by cell death ELISA (enzyme-linked immunosorbent assay), an increase in the number of TUNEL (terminal dUTP nick-end labeling)-positive cells and a reduction in the mitochondrial metabolism of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). No significant effect was observed with the release of lactate dehydrogenase (LDH), indicating that cell death presumably occurred via apoptotic mechanisms. Co-incubation of MPP+ with L-carnitine significantly reduced MPP+-induced apoptosis. Western blot analyses showed that neurotoxic concentrations of MPP+ decreased the ratio of BCL-X(L) to Bax and decreased the protein levels of polysialic acid neural cell adhesion molecules (PSA-NCAM), a neuron specific marker. L-carnitine blocked these effects of MPP+ suggesting its potential therapeutic utility in degenerative disorders such as Parkinson's disease, Alzheimer's disease, ornithine transcarbamylase deficiency and other mitochondrial diseases. PMID- 17084540 TI - Extracranial chordoma: Outcome in patients treated with function-preserving surgery followed by spot-scanning proton beam irradiation. AB - PURPOSE: To evaluate the use of postoperative proton therapy (PT) in extracranial chordoma. PATIENTS AND METHODS: Twenty-six patients were treated. Gross total resection was achieved in 18 patients. Nine patients had cervical, 2 had thoracic, 8 had lumbar, and 7 had sacro-coccygeal chordomas. Thirteen patients had implants. PT was administered after function-preserving surgery, using a gantry and spot scanning, without or with intensity modulation (IMPT; 6 patients), and/or photon-based radiotherapy (RT, 6 patients). Median total dose was 72 cobalt Gray equivalent (CGE; range, 59.4-74.4), with means of 70.5 and 73.2 CGE for patients with and without implants. Median follow-up time was 35 months (range, 13-73 months). Adverse events were scored using the Common Terminology Criteria for Adverse Events grading system (version 3.0). RESULTS: At 3 years, actuarial overall survival (OS) and progression-free survival (PFS) rates were 84% and 77%, respectively. One patient each died of local failure (LF), distant failure (DF), suicide, and secondary tumor. We observed 5 LFs and 3 DFs; 3-year LF-free and DF-free survival rates were 86%. We observed four radiation-induced late adverse events (Grade 2 sensory neuropathy; Grade 3 subcutaneous necrosis, and osteonecrosis; and Grade 5 secondary cancer). In univariate analysis, implants were associated with LF (p = 0.034). Gross residual tumor above 30 mL was negatively associated with OS (p = 0.013) and PFS (p = 0.025). CONCLUSIONS: Postoperative PT for extracranial chordomas delivered with spot scanning offers high local control rates. Toxicity was acceptable. Implants were significantly associated with LF. Residual tumor above 30 mL impacted negatively on OS and PFS. PMID- 17084541 TI - Intensity-modulated radiotherapy for pituitary adenomas: the preliminary report of the Cleveland Clinic experience. AB - PURPOSE: Intensity-modulated radiotherapy (IMRT) is being increasingly used for the treatment of pituitary adenomas. However, there have been few published data on the short- and long-term outcomes of this treatment. This is the initial report of the Cleveland Clinic's experience. METHODS AND MATERIALS: Between February 1998 and December 2003, 34 patients with pituitary adenomas were treated with IMRT. A retrospective chart review was conducted for data analysis. RESULTS: With a median follow-up of 42.5 months, the treatment has proven to be well tolerated, with performance status remaining stable in 90% of patients. Radiographic local control was 89%, and among patients with secretory tumors, 100% had a biochemical response. Only 1 patient required salvage surgery for progressive disease, giving a clinical progression free survival of 97%. The only patient who received more than 46 Gy experienced optic neuropathy 8 months after radiation. Smaller tumor volume significantly correlated with subjective improvements in nonvisual neurologic complaints (p = 0.03), and larger tumor volume significantly correlated with subjective worsening of visual symptoms (p = 0.05). New hormonal supplementation was required for 40% of patients. Younger patients were significantly more likely to require hormonal supplementation (p = 0.03). CONCLUSIONS: Intensity-modulated radiation therapy is a safe and effective treatment for pituitary adenomas over the short term. Longer follow-up is necessary to determine if IMRT confers any advantage with respect to either tumor control or toxicity over conventional radiation modalities. PMID- 17084542 TI - Adjuvant paclitaxel and carboplatin chemotherapy with involved field radiation in advanced endometrial cancer: a sequential approach. AB - PURPOSE: To determine the feasibility of adjuvant paclitaxel and carboplatin chemotherapy interposed with involved field radiotherapy for women with advanced endometrial cancer. METHODS AND MATERIALS: This was a prospective cohort study of women with Stage III and IV endometrial cancer. Adjuvant therapy consisted of 4 cycles of paclitaxel (175 mg/m(2)) and carboplatin (350 mg/m(2)) every 3 weeks, followed sequentially by external beam radiotherapy (RT) to the pelvis (45 Gy), followed by an additional two cycles of chemotherapy. Para-aortic RT and/or HDR vault brachytherapy (BT) were added at the discretion of the treating physician. RESULTS: Thirty-three patients (median age, 63 years) received treatment between April 2002 and June 2005. Median follow-up was 21 months. Stage distribution was as follows: IIIA (21%), IIIC (70%), IVB (9%). Combination chemotherapy was successfully administered to 30 patients (91%) and 25 patients (76%), before and after RT respectively. Nine patients (27%) experienced acute Grade 3 or 4 chemotherapy toxicities. All patients completed pelvic RT; 19 (58%) received standard 4-field RT and 14 (42%) received intensity-modulated radiotherapy. Ten (30%) received extended field radiation. Four patients (12%) experienced acute Grade 3 or 4 RT toxicities. Six (18%) patients developed chronic RT toxicity. There were no treatment-related deaths. Two-year disease-free and overall survival rates were both 55%. There was only one pelvic relapse (3%). CONCLUSIONS: Adjuvant treatment with combination chemotherapy interposed with involved field radiation in advanced endometrial cancer was well tolerated. This protocol may be suitable for further evaluation in a clinical trial. PMID- 17084543 TI - Intraoperative radiation therapy for recurrent head-and-neck cancer: the UCSF experience. AB - PURPOSE: To review a single-institutional experience with the use of intraoperative radiation therapy (IORT) for recurrent head-and-neck cancer. METHODS AND MATERIALS: Between 1991 and 2004, 137 patients were treated with gross total resection and IORT for recurrence or persistence of locoregional cancer of the head and neck. One hundred and thirteen patients (83%) had previously received external beam radiation as a component of definitive therapy. Ninety-four patients (69%) had squamous cell histology. Final surgical margins were microscopically positive in 56 patients (41%). IORT was delivered using either a modified linear accelerator or a mobile electron unit and was administered as a single fraction to a median dose of 15 Gy (range, 10-18 Gy). Median follow-up among surviving patients was 41 months (range, 3-122 months). RESULTS: The 1-year, 2-year, and 3-year estimates of in-field control after salvage surgery and IORT were 70%, 64%, and 61%, respectively. Positive margins at the time of IORT predicted for in-field failure (p = 0.001). The 3-year rates of locoregional control, distant metastasis-free survival, and overall survival were 51%, 46%, and 36%, respectively. There were no perioperative fatalities. Complications included wound infection (4 patients), orocutaneous fistula (2 patients), flap necrosis (1 patient), trismus (1 patient), and neuropathy (1 patient). CONCLUSIONS: Intraoperative RT results in effective disease control with acceptable toxicity and should be considered for selected patients with recurrent or persistent cancers of the head and neck. PMID- 17084544 TI - 15-Year biochemical relapse free survival in clinical Stage T1-T3 prostate cancer following combined external beam radiotherapy and brachytherapy; Seattle experience. AB - PURPOSE: Long-term biochemical relapse-free survival (BRFS) rates in patients with clinical Stages T1-T3 prostate cancer continue to be scrutinized after treatment with external beam radiation therapy and brachytherapy. METHODS AND MATERIALS: We report 15-year BRFS rates on 223 patients with clinically localized prostate cancer that were consecutively treated with I(125) or Pd (103) brachytherapy after 45-Gy neoadjuvant EBRT. Multivariate regression analysis was used to create a pretreatment clinical prognostic risk model using a modified American Society for Therapeutic Radiology and Oncology consensus definition (two consecutive serum prostate-specific antigen rises) as the outcome. Gleason scoring was performed by the pathologists at a community hospital. Time to biochemical failure was calculated and compared by using Kaplan-Meier plots. RESULTS: Fifteen-year BRFS for the entire treatment group was 74%. BRFS using the Memorial Sloan-Kettering risk cohort analysis (95% confidence interval): low risk, 88%, intermediate risk 80%, and high risk 53%. Grouping by the risk classification described by D'Amico, the BRFS was: low risk 85.8%, intermediate risk 80.3%, and high risk 67.8% (p = 0.002). CONCLUSIONS: I(125) or Pd(103) brachytherapy combined with supplemental EBRT results in excellent 15-year biochemical control. Different risk group classification schemes lead to different BRFS results in the high-risk group cohorts. PMID- 17084545 TI - Retroperitoneal soft tissue sarcoma: an analysis of radiation and surgical treatment. AB - PURPOSE: To evaluate the clinical outcomes of patients with localized retroperitoneal soft tissue sarcoma (STS) treated with complete surgical resection and radiation. METHODS AND MATERIALS: The medical records of 83 patients were reviewed retrospectively. Sixty patients presented with primary disease and the remaining 23 had recurrence after previous surgical resection. RESULTS: With a median follow-up of 47 months, the actuarial overall disease specific survival (DSS), distant metastasis-free survival, and local control (LC) rates were 44%, 67%, and 40%, respectively. Of the 38 patients dying of disease, local disease progression was the sole site of recurrence for 16 patients and was a component of progression for another 11 patients. Multivariate analysis indicated that histologic grade was associated with the 5-year rates of DSS (low grade, 92%; intermediate-grade, 51%; and high-grade, 41%, p = 0.006). Multivariate analysis also indicated an inferior 5-year LC rate for patients presenting with recurrent disease, positive or uncertain resection margins, and age greater than 65 years. The data did not suggest an improved local control with higher doses of external-beam radiation (EBRT) or with the specific use of intraoperative radiotherapy (IORT). Radiation-related complications (10% at 5 years) developed in 5 patients; all had received their EBRT postoperatively. CONCLUSIONS: Although preoperative radiation therapy and aggressive surgical resection is well tolerated in patients, local disease progression continues to be a significant component of disease death. In this small cohort of patients, the use of higher doses of EBRT or IORT did not result in clinically apparent improvements in outcomes. PMID- 17084546 TI - A magnetic resonance imaging study of prostate deformation relative to implanted gold fiducial markers. AB - PURPOSE: To describe prostate deformation during radiotherapy and determine the margins required to account for prostate deformation after setup to intraprostatic fiducial markers (FM). METHODS AND MATERIALS: Twenty-five patients with T1c-T2c prostate cancer had three gold FMs implanted. The patients presented with a full bladder and empty rectum for two axial magnetic resonance imaging (MRI) scans using a gradient recalled echo (GRE) sequence capable of imaging the FMs. The MRIs were done at the time of radiotherapy (RT) planning and a randomly assigned fraction. A single observer contoured the prostate surfaces. They were entered into a finite element model and aligned using the centroid of the three FMs. RESULTS: During RT, the prostate volume decreased by 0.5%/fraction (p = 0.03) and the FMs in-migrated by 0.05 mm/fraction (p < 0.05). Prostate deformation was unrelated to differential bladder and bowel filling, but was related to a transurethral resection of the prostate (TURP) (p = 0.003). The standard deviation for systematic uncertainty of prostate surface contouring was 0.8 mm and for FM centroid localization was 0.4 mm. The standard deviation of random interfraction prostate deformation was 1.5 mm and for FM centroid variability was 1.1 mm. These uncertainties from prostate deformation can be incorporated into a margin recipe to determine the total margins required for RT. CONCLUSIONS: During RT, the prostate exhibited: volume decrease, deformation, and in-migration of FMs. Patients with TURPs were prone to prostate deformation. PMID- 17084547 TI - Long-term results of endobronchial brachytherapy: A curative treatment? AB - PURPOSE: To evaluate outcomes after high-dose-rate endobronchial brachytherapy (HDR-EBBT) for limited lung carcinoma. METHODS: A total of 106 patients with endobronchial lung cancer and not eligible for surgery or external beam radiotherapy, without nodal or visceral metastases, were treated with HDR-EBBT. They had developed disease relapse after surgery (n = 43) or external beam radiotherapy (n = 27) or had early lung cancer with respiratory insufficiency (n = 36). Treatment consisted of six fractions of 5 or 7 Gy, usually delivered 1 cm from the source. RESULTS: The complete histologic response rate, evaluated at 3 months after HDR-EBBT, was 59.4%. At 3 and 5 years, the local control, overall survival, and cause-specific survival rates were 60.3% and 51.6%, 47.4 and 24%, and 67.9 and 48.5%, respectively. Factors significantly associated with local failure were high tumor volume (tumor length >2 cm, bronchial obstruction >25%, tumor visibility on CT scan) and previous endoscopic treatment. Cause-specific survival, but not overall survival, was significantly associated with local control, probably because of the high rate of deaths not related to lung cancer. Five deaths were attributed to the HDR-EBBT procedure (two from fatal hemoptysis and three from bronchial necrosis). CONCLUSION: High-dose-rate-EBBT achieved a long-term cause-specific survival rate of 50% of the patients with localized endobronchial carcinoma and could be considered curative. PMID- 17084548 TI - Dosimetric and toxicity comparison between prone and supine position IMRT for endometrial cancer. AB - PURPOSE: To determine the dosimetric and toxicity differences between prone and supine position intensity-modulate radiotherapy in endometrial cancer patients treated with adjuvant radiotherapy. METHODS: Forty-seven consecutive endometrial cancer patients treated with adjuvant RT were analyzed. Of these, 21 were treated in prone position and 26 in the supine position. Dose-volume histograms for normal tissue structures and targets were compared between the two groups. Acute and chronic toxicity were also compared between the cohorts. RESULTS: The percentage of volume receiving 10, 20, 30, 40, 45, and 50 Gy for small bowel was 89.5%, 69%, 33%, 12.2%, 5%, and 0% in the prone group and 87.5%, 62.7%, 26.4%, 8%, 4.3%, and 0% in the supine group, respectively. The difference was not statistically significant. The dose-volume histograms for bladder and rectum were also comparable, except for a slightly greater percentage of volume receiving 10 Gy (1.5%) and 20 Gy (5%) for the rectum in the prone group. Acute small bowel toxicities were Grade 1 in 7 patients and Grade 2 in 14 patients in the prone group vs. Grade 1 in 6 patients and Grade 2 in 19 patients in the supine group. Chronic toxicity was Grade 1 in 7 patients and Grade 3 in 1 patient in the prone group and Grade 1 in 5 patients in the supine group. CONCLUSION: These preliminary results suggest that no difference exists in the dose to the normal tissue and toxicity between prone and supine intensity-modulated radiotherapy for endometrial cancer. Longer follow-up and more outcome studies are needed to determine whether any differences exist between the two approaches. PMID- 17084549 TI - A Phase II study of acute toxicity for Celebrex (celecoxib) and chemoradiation in patients with locally advanced cervical cancer: primary endpoint analysis of RTOG 0128. AB - PURPOSE: To determine treatment-related acute toxicity rates in patients with locally advanced cervical cancer treated by oral celecoxib, i.v. cisplatin and 5 FU, and concurrent pelvic radiation therapy. METHODS AND MATERIALS: Eligible patients on this RTOG Phase I-II study for advanced cervix cancer included FIGO Stage IIB-IVA or patients with FIGO Stage IB through IIA with biopsy proven pelvic node metastases or tumor size > or =5 cm. Patients were treated with pelvic radiotherapy and brachytherapy. Celecoxib was prescribed at 400 mg twice daily beginning on day 1 for 1 year. Cisplatin (75 mg/m2) and 5-FU (1g/m2 for 4 days) were administered every 3 weeks times 3. The primary end point of the study was treatment related toxicity. RESULTS: Between August 2001 and March 2004, 84 patients were accrued to the study and 77 patients were evaluable for toxicity. Regarding the primary end point, toxicities were observed in the following areas: blood/bone marrow (16), gastrointestinal (14), pain (7), renal/genitourinary (6), cardiovascular (3), hemorrhage (1), and neurologic (1). For the first 75 evaluable patients, a toxicity failure was identified in 36 patients for a rate of 48%. CONCLUSIONS: Celecoxib at 400 mg twice daily together with concurrent cisplatin and 5-FU and pelvic radiotherapy has a high incidence of acute toxicities. The most frequent toxicities were hematologic. Albeit, the toxicity was deemed excessive in this trial, the rate of toxicities was not too different compared to other recent experiences with concurrent chemoradiation for advanced cervix cancer. PMID- 17084550 TI - Combination ibandronate and radiotherapy for the treatment of bone metastases: clinical evaluation and radiologic assessment. AB - PURPOSE: Ibandronate is a single-nitrogen, noncyclic bisphosphonate with proven efficacy for reducing metastatic bone pain. In this study, we assessed the palliative effects of combined ibandronate and radiotherapy. METHODS AND MATERIALS: Forty-five patients with bone metastases from various solid tumors received external-beam radiotherapy, 30-40 Gy over 3-4.5, weeks combined with 10 cycles of monthly intravenous ibandronate, 6 mg. RESULTS: After combined therapy, mean bone pain scores (graded from 0 to 10) were reduced from 6.3 at baseline to 0.8 after 3 months, with further reductions at later time points (all p < 0.001). Opioid use decreased from 84% of patients at baseline (38/45) to 24% (11/45) at 3 months, with further subsequent reductions (all p < 0.001). Mean performance status and functioning scores also significantly improved. Bone density (assessed by computed tomography scan) increased by 20% vs. baseline at 3 months, 46% at 6 months, and 73% at 10 months (all p < 0.001). Lesion improvement was also demonstrated by magnetic resonance imaging. Treatment was well tolerated with no renal toxicity. CONCLUSIONS: In this pilot study, combined radiotherapy and ibandronate provided substantial bone pain relief and increased bone density. Computed tomography-based or magnetic resonance imaging-based evaluations offer objective methods for assessing therapeutic outcomes. PMID- 17084551 TI - Results of a phase II trial of transrectal ultrasound-guided permanent radioactive implantation of the prostate for definitive management of localized adenocarcinoma of the prostate (radiation therapy oncology group 98-05). AB - PURPOSE: To evaluate the effectiveness of transrectal ultrasound-guided permanent radioactive (125)I implantation of the prostate for organ-confined adenocarcinoma of the prostate compared with historical data of prostatectomy and external beam radiotherapy within a cooperative group setting. METHODS AND MATERIALS: Patients accrued to this study had histologically confirmed, locally confined, adenocarcinoma of the prostate with clinical Stage T1b, T1c, or T2a, no nodal or metastatic disease, prostate-specific antigen level of < or =10 ng/mL, and Gleason score of < or =6. All patients underwent transrectal ultrasound-guided radioactive (125)I permanent seed implantation into the prostate. The prescribed dose was 145 Gy to the prostate planning target volume. RESULTS: A total of 27 institutions accrued a total of 101 patients to this protocol, with no institution accruing >8 patients. Six patients were ineligible, leaving 95 properly entered as eligible in the study. The median follow-up was 5.3 years (range, 0.4-6.5 years). At 5 years, 5 patients had local failure, 1 had evidence of distant failure, and 6 (6%) had biochemical failure. The overall survival rate at 5 years was 96.7%. At last follow-up, no patient had died of prostate cancer or related toxicities. Eight patients had a maximal acute toxicity level of 3, and no patient had Grade 4 or 5 acute toxicity. During follow-up, 2 patients had maximal Grade 3 toxicity, both related to bladder issues, and no patient experienced Grade 4 or 5 toxicity. CONCLUSION: The results of this clinical protocol (a multi-institutional trial of brachytherapy for localized adenocarcinoma of the prostate) have demonstrated that this type of trial can be successfully completed through the Radiation Therapy Oncology Group. Biochemical disease-free survival was comparable with other brachytherapy published series and with the results after surgery and external beam radiotherapy. PMID- 17084552 TI - Daoy medulloblastoma cells that express CD133 are radioresistant relative to CD133- cells, and the CD133+ sector is enlarged by hypoxia. AB - PURPOSE: Primary medulloblastoma and glioblastoma multiforme tumor cells that express the surface marker CD133 are believed to be enriched for brain tumor stem cells because of their unique ability to initiate or reconstitute tumors in immunodeficient mice. This study sought to characterize the radiobiological properties and marker expression changes of CD133+ vs. CD133- cells of an established medulloblastoma cell line. METHODS AND MATERIALS: Daoy and D283 Med cell lines were stained with fluorescently labeled anti-CD133 antibody and sorted into CD133+ and CD133- populations. The effect of oxygen (2% vs. 20%) on CD133 expression was measured. Both populations were analyzed for marker stability, cell cycle distribution, and radiosensitivity. RESULTS: CD133+ Daoy cells restored nearly native CD133+ and CD133- populations within 18 days, whereas CD133- cells remained overwhelmingly CD133-. Culturing Daoy cells in 2% oxygen rather than the standard 20% oxygen increased their CD133 expression 1.6-fold. CD133+ Daoy cells were radioresistant via the beta-parameter of the linear quadratic model relative to CD133- Daoy cells, although their alpha-parameters and cell cycle distributions were identical. CONCLUSIONS: Restoration of the original CD133+ and CD133- populations from CD133+ Daoy cells in serum is further evidence that CD133+ cells are functionally distinct from CD133- cells. The radioresistance of CD133+ compared with CD133- Daoy cells is consistent with better repair of sublethal damage. Enlargement of the CD133+ sector is a new feature of the hypoxic response. PMID- 17084553 TI - Permanent prostate implant using high activity seeds and inverse planning with fast simulated annealing algorithm: A 12-year Canadian experience. AB - PURPOSE: To report outcomes and toxicity of the first Canadian permanent prostate implant program. METHODS AND MATERIALS: 396 consecutive patients (Gleason < or =6, initial prostate specific antigen (PSA) < or =10 and stage T1-T2a disease) were implanted between June 1994 and December 2001. The median follow-up is of 60 months (maximum, 136 months). All patients were planned with fast-simulated annealing inverse planning algorithm with high activity seeds ([gt] 0.76 U). Acute and late toxicity is reported for the first 213 patients using a modified RTOG toxicity scale. The Kaplan-Meier biochemical failure-free survival (bFFS) is reported according to the ASTRO and Houston definitions. RESULTS: The bFFS at 60 months was of 88.5% (90.5%) according to the ASTRO (Houston) definition and, of 91.4% (94.6%) in the low risk group (initial PSA < or =10 and Gleason < or =6 and Stage < or =T2a). Risk factors statistically associated with bFFS were: initial PSA >10, a Gleason score of 7-8, and stage T2b-T3. The mean D90 was of 151 +/- 36.1 Gy. The mean V100 was of 85.4 +/- 8.5% with a mean V150 of 60.1 +/- 12.3%. Overall, the implants were well tolerated. In the first 6 months, 31.5% of the patients were free of genitourinary symptoms (GUs), 12.7% had Grade 3 GUs; 91.6% were free of gastrointestinal symptoms (GIs). After 6 months, 54.0% were GUs free, 1.4% had Grade 3 GUs; 95.8% were GIs free. CONCLUSION: The inverse planning with fast simulated annealing and high activity seeds gives a 5-year bFFS, which is comparable with the best published series with a low toxicity profile. PMID- 17084554 TI - Thyroid disorders in patients treated with radiotherapy for head-and-neck cancer: a retrospective analysis of seventy-three patients. AB - PURPOSE: To evaluate the incidence of thyroid disorders and dose distribution to the thyroid in patients treated with radiotherapy for head-and-neck carcinomas. METHODS AND MATERIALS: A retrospective evaluation of data from 73 patients treated for head-and-neck cancers in our department was performed. Thyroid function was evaluated mainly by the measurement of thyrotropin (thyroid stimulating hormone [TSH]). A retrospective analysis of treatment plans was performed for 57 patients. Percentages of thyroid glandular volume absorbing 10, 30, and 50 Gy (V10, V30, and V50 respectively) were considered for statistical analysis. RESULTS: A majority of patients (61%) had a normal thyroid function whereas 19 patients (26%) had hypothyroidism. Mean thyroid volume was 30.39 cc. Point 3 (located at isthmus) absorbed lower doses compared with other points (p < 0.0001). Median values of V10, V30, and V50 were 92% (range, 57-100%), 75% (range, 28.5-100%), and 35% (range, 3-83%) respectively. Gender was associated with toxicity (presence of any kind of thyroid disorders) (p < 0.05), with females displaying higher levels of TSHr (relative TSH = patient's value/maximum value of the laboratory range) (p = 0.0005) and smaller thyroid volume (p = 0.0012) compared with male population. TSHr values were associated with thyroid volume, and the presence of midline shielding block in the anterior field was associated with relative free thyroxine (FT4r = patient's value/maximum value of the laboratory range) values. CONCLUSIONS: Gender and thyroid volume seem to play an important role in the occurrence of thyroid toxicity, but further studies on dose-effect relationship for radiotherapy-induced thyroid toxicity are needed. PMID- 17084555 TI - A comparative study of volumetric analysis, histopathologic downstaging, and tumor regression grade in evaluating tumor response in locally advanced rectal cancer following preoperative chemoradiation. AB - PURPOSE: To compare tumor volume reduction rate, histopathologic downstaging, and tumor regression grade (TRG) among tumor responses in rectal cancer after preoperative chemoradiotherapy (CRT). PATIENTS AND METHODS: Between 2002 and 2004, 30 patients with locally advanced rectal cancer underwent preoperative CRT, followed by surgical resection. Magnetic resonance volumetry was performed before and after CRT. Histopathologic tumor staging and tumor regression were reviewed. We compared pre- and post-CRT tumor volume and percent of volume reduction, according to histopathologic downstaging and TRG. RESULTS: The tumor volume reduction rates ranged from 14.6% to 100%. Mean pre- and post-CRT tumor volumes were significantly smaller in patients who showed T downstaging than in those who did not (p = 0.040, 0.014). The mean tumor volume reduction was 66.4% vs. 55.2% (p = 0.361). However, the mean pre- and post-CRT tumor volume and mean tumor volume reduction rate between patients who showed N downstaging and those who did not were not statistically different (p = 0.176, 0.767, and 0.899). With respect to TRG, the mean pre- and post-CRT tumor volumes were not statistically significant (p = 0.108, 0.708, and 0.120). CONCLUSION: Tumor volume reduction rate does not correlate with histopathologic downstaging and TRG. It might be hazardous to evaluate tumor response with respect to volume reduction and to select the surgical method on this basis. PMID- 17084556 TI - Intensity modulated radiation-therapy for preoperative posterior abdominal wall irradiation of retroperitoneal liposarcomas. AB - PURPOSE: Preoperative external-beam radiation therapy (preop RT) in the management of Retroperitoneal Liposarcomas (RPLS) typically involves the delivery of radiation to the entire tumor mass: yet this may not be necessary. The purpose of this study is to evaluate a new strategy of preop RT for RPLS in which the target volume is limited to the contact area between the tumoral mass and the posterior abdominal wall. METHODS AND MATERIALS: Between June 2000 and Jan 2005, 18 patients with the diagnosis of RPLS have been treated following a pilot protocol of pre-op RT, 50 Gy in 25 fractions of 2 Gy/day. The Clinical Target Volume (CTV) has been limited to the posterior abdominal wall, region at higher risk for local relapse. A Three-Dimensional conformal (3D-CRT) and an Intensity Modulated (IMRT) plan were generated and compared; toxicity was reported following the National Cancer Institute (NCI) Common Terminology Criteria for Adverse Events v3.0. RESULTS: All patients completed the planned treatment and the acute toxicity was tolerable: 2 patients experienced Grade 3 and 1 Grade 2 anorexia while 2 patients developed Grade 2 nausea. IMRT allows a better sparing of the ipsilateral and the contralateral kidney. All tumors were successfully resected without major complications. At a median follow-up of 27 months 2 patients developed a local relapse and 1 lung metastasis. CONCLUSIONS: Our strategy of preop RT is feasible and well tolerated: the rate of resectability is not compromised by limiting the preop CTV to the posterior abdominal wall and a better critical-structures sparing is obtained with IMRT. PMID- 17084557 TI - Spot-scanning proton therapy for malignant soft tissue tumors in childhood: First experiences at the Paul Scherrer Institute. AB - PURPOSE: Radiotherapy plays a major role in the treatment strategy of childhood sarcomas. Consequences of treatment are likely to affect the survivor's quality of life significantly. We investigated the feasibility of spot-scanning proton therapy (PT) for soft tissue tumors in childhood. METHODS AND MATERIALS: Sixteen children with soft tissue sarcomas were included. Median age at PT was 3.3 years. In 10 children the tumor histology was embryonal rhabdomyosarcoma. All tumors were located in the head or neck, parameningeal, or paraspinal, or pelvic region. In the majority of children, the tumor was initially unresectable (Intergroup Rhabdomyosarcoma Study [IRS] Group III in 75%). In 50% of children the tumors exceeded 5 cm. Fourteen children had chemotherapy before and during PT. Median total dose of radiotherapy was 50 cobalt Gray equivalent (CGE). All 16 children were treated with spot-scanning proton therapy at the Paul Scherrer Institute, and in 3 children the PT was intensity-modulated (IMPT). RESULTS: After median follow-up of 1.5 years, local control was achieved in 12 children. Four children failed locally, 1 at the border of the radiation field and 3 within the field. All 4 children died of tumor recurrence. All 4 showed unfavorable characteristic either of site or histopathology of the tumor. Acute toxicity was low, with Grade 3 or 4 side effects according to Radiation Therapy Oncology Group/European Organization for Research and Treatment of Cancer (RTOG/EORTC) criteria occurring in the bone marrow only. CONCLUSIONS: Proton therapy was feasible and well tolerated. Early local control rates are comparable to those being achieved after conventional radiotherapy. For investigations on late effect, longer follow-up is needed. PMID- 17084558 TI - Multi-institutional analysis of long-term outcome for stages T1-T2 prostate cancer treated with permanent seed implantation. AB - PURPOSE: To assess long-term prostate-specific antigen (PSA) outcome after permanent prostate brachytherapy (BT) and identify predictors of improved disease free survival. METHODS AND MATERIALS: Eleven institutions combined data on 2,693 patients treated with permanent interstitial BT monotherapy for T1-T2 prostate cancer. Of these patients, 1,831 (68%) were treated with I-125 (median dose, 144 Gy) and 862 (32%) were treated with Pd-103 (median dose, 130 Gy). Criteria for inclusion were: available pre-BT PSA, BT > or =5 years before data submission, BT between 1988-1998, and no androgen deprivation before failure. The median follow up was 63 months. RESULTS: Among patients where the I-125 dose to 90% of the prostate (D90) was > or =130 Gy, the 8-year PSA relapse-free survival (PRFS) was 93% compared with 76% for those with lower D90 dose levels (p < 0.001). A multivariable analysis identified tumor stage (p = 0.002), Gleason score (p < 0.001), pretreatment PSA level (p < 0.001), treatment year (p = 0.001), and the isotope used (p = 0.004) as pretreatment and treatment variables associated with PRFS. When restricted to patients with available postimplantation dosimetric information, D90 emerged as a significant predictor of biochemical outcome (p = 0.01), and isotope was not significant. The 8-year PRFS was 92%, 86%, 79%, and 67%, respectively, for patients with PSA nadir values of 0-0.49, 0.5-0.99, 1.0 1.99, and >2.0 ng/mL (p < 0.001). Among patients free of biochemical relapse at 8 years, the median nadir level was 0.1 ng/mL, and 90% of these patients achieved a nadir PSA level <0.6 ng/mL. CONCLUSIONS: Outcome after permanent BT for prostatic cancer relates to tumor stage, Gleason score, pretreatment PSA, BT year, and post BT dosimetric quality. PSA nadir < or =0.5 ng/mL was particularly associated with durable long-term PSA disease-free survival. The only controllable factor to impact on long-term outcome was the D90 which is a reflection of implant quality. PMID- 17084559 TI - p53 status is a major determinant of effects of decreasing peroxiredoxin I expression on tumor growth and response of lung cancer cells to treatment. AB - PURPOSE: The potential roles of peroxiredoxin (Prx) I in carcinogenesis and treatment have been explored. Our previous study revealed differences between A549 (functional p53) and H1299 (null p53) Prx I antisense transfectants. The discrepancy might have resulted from the p53 status. In this study, we further investigated the role of Prx I and p53 on lung cancer growth and the response to treatment in vitro and in vivo. METHODS: We established stable A549 and H1299 transfectants with Prx I antisense and p53, respectively. We then examined their characteristics in vitro and used nude mice xenografts of these cell lines to compare their capacity for tumor invasion and spontaneous metastasis and their sensitivity to radiotherapy. RESULTS: Increased reactive oxygen species caused by lower Prx I activity induced p53 expression. In lethal stress, the augmentation of reactive oxygen species was partially reversed by blocking p53 in A549 with Prx I antisense. We demonstrated the potential contribution of p53-dependent mechanisms to inhibit lung tumor growth and increase radiosensitization using H1299 transfected with p53 in vitro and in vivo. An increased p53 level attenuated the capacity of the cells for metastasis by decreasing vascular endothelial growth factor and induced radiosensitization by increased apoptosis and cell senescence and by regulating intracellular reactive oxygen species. CONCLUSION: These results suggest that p53 status has an important role in the tumor-inhibiting and radiosensitizing effects of decreasing Prx I. Both Prx I and p53 may be powerful prognosticators for lung cancer. PMID- 17084560 TI - Update of human spinal cord reirradiation tolerance based on additional data from 38 patients. AB - PURPOSE: To update a combined analysis of all published clinical data. METHODS AND MATERIALS: We collected data from 38 additional patients treated in our department or published in four different reports and calculated the biologically effective dose (BED) according to the linear-quadratic model using an alpha/beta value of 2 Gy for cervical and thoracic cord and 4 Gy for lumbar cord. In this model, a dose of 50 Gy given in single daily fractions of 2 Gy is equivalent to a BED of 100 Gy(2) or 75 Gy(4). RESULTS: The 2005 risk score based on three variables (cumulative BED, highest BED of all treatment series in a particular individual, and interval), which discriminate three different risk groups, does not require modification. The low-risk group now contains 1 case of radiation myelopathy (RM) after hypofractionated stereotactic reirradiation. Therefore, the rate increased from 0% to 3%. Intermediate-risk patients developed RM in 25%, and high-risk patients in 90%. When the interval between the two treatment courses is not shorter than 6 months and the dose of each course is < or =98 Gy(2), the cumulative BED where no case of RM has yet been reported is 120 Gy(2). CONCLUSIONS: Based on these updated results, the risk of RM appears small after < or =135.5 Gy(2) when the interval is not shorter than 6 months and the dose of each course is < or =98 Gy(2). We would recommend limiting the dose to the lowest feasible level. The influence of very steep dose gradients from stereotactic and intensity-modulated approaches (i.e., a more complex volume-effect) requires further evaluation. PMID- 17084561 TI - Relation between acute and late irradiation impairment of four basic tastes and irradiated tongue volume in patients with head-and-neck cancer. AB - PURPOSE: Taste loss is a major cause of morbidity in patients undergoing head-and neck irradiation. The relationship between the time course and the degree of taste disorder was studied in both acute and late phases. METHODS AND MATERIALS: Taste ability was measured by the taste threshold for the four basic tastes using a filter paper disc method in patients before, during, and after radiotherapy. The subjects were divided into two groups. In Group A, the radiation fields included most of the tongue (n = 100), and in Group B the radiation fields did not include the tip of the tongue (n = 18). RESULTS: In Group A, there was a significant impairment of the threshold of all four basic tastes at 3 weeks after starting radiotherapy (RT), and this impairment remained at 8 weeks (p < 0.05). This was not seen in Group B. In Group A, there was no significant difference in the patterns of taste sensitivity change between the high-dose (>20 Gy) and low dose (< or =20 Gy) groups. In the late phase, recovery of taste loss was seen in both groups since 4 months after completing RT. CONCLUSIONS: Unless the anterior part of the tongue was irradiated, taste loss was not observed during RT. When the anterior part of the tongue was irradiated, a difference by radiation dose was not observed in the taste loss pattern. Additionally, radiation-induced taste dysfunction appears to be a temporal effect. PMID- 17084562 TI - Targeted intraoperative radiotherapy (TARGIT) yields very low recurrence rates when given as a boost. AB - PURPOSE: Patients undergoing breast-conserving surgery were offered boost radiotherapy with targeted intraoperative radiotherapy (TARGIT) using the Intrabeam system to test the feasibility, safety, and efficacy of the new approach. METHODS AND MATERIALS: We treated 302 cancers in 301 unselected patients. This was not a low-risk group. One-third of patients (98/301) were younger than 51 years of age. More than half of the tumors (172, 57%) were between 1 cm and 2 cm, and one-fifth (62, 21%) were >2 cm; 29% (86) had a Grade 3 tumor and, in 29% (87), axillary lymph nodes contained metastasis. After primary surgery, 20 Gy was delivered intraoperatively to the surface of the tumor bed, followed by external-beam radiotherapy (EBRT), but excluding the usual boost. RESULTS: The treatment was well tolerated. The follow-up ranged from 3 to 80 months (164 and 90 patients completed 2 and 3 years follow-up, respectively). Four patients (1.3%) had local recurrence. The Kaplan-Meier estimate of local recurrence is 2.6% (SE = 1.7) at 5 years. This compares favorably with the 4.3% recurrence rate in boosted patients from the EORTC boost study, in which only 8.1% patients were node-positive, as opposed to 29% in our series. CONCLUSION: Targeted intraoperative radiotherapy combined with EBRT results in a low local recurrence rate. This could be attributed to both accurate targeting and timeliness of the treatment. These data support the need for a randomized trial to test whether the TARGIT boost is superior to conventional external boost, especially in high-risk women. PMID- 17084563 TI - Does gender contribute to heterogeneity in criteria for cannabis abuse and dependence? Results from the national epidemiological survey on alcohol and related conditions. AB - Previous research has noted that a unidimensional latent construct underlies criteria for cannabis abuse and dependence. However, no study to date has explored whether gender contributes to heterogeneity in the latent abuse and dependence construct and furthermore, whether after accounting for differences in the mean scores of abuse and dependence across genders, there is any evidence for heterogeneity in the individual abuse and dependence criteria. The present study utilizes data on criteria for cannabis abuse and dependence from a large, nationally representative sample (National Epidemiological Survey on Alcohol and Related Conditions) of 8172 lifetime cannabis users to investigate whether gender contributes to heterogeneity in the underlying construct of cannabis abuse and dependence, and in each individual criterion as well. Analyses, all of which were conducted in MPlus, included factor analysis, as well as MIMIC and multiple-group models for an examination of dimensionality and gender heterogeneity, respectively. Results favor a unidimensional construct for cannabis abuse/dependence, as seen in prior research. We also identify two abuse (legal and hazard) and two dependence (quit and problems) criteria, which show significant gender heterogeneity with the abuse criteria exhibiting higher thresholds in women and the dependence criteria in men. We conclude that the criteria that serve as indicators of DSM-IV cannabis abuse and dependence do not function identically in men and women and that certain criteria (e.g. hazardous use) require further refinement. PMID- 17084564 TI - Monoclonal antibodies reveal a weak interaction between the F18 fimbrial adhesin FedF and the major subunit FedA. AB - F18+ Escherichia coli can cause post-weaning diarrhoea and oedema disease in pigs. These diseases are responsible for substantial economic losses, but a vaccine is not available. A good knowledge of the characteristic of the fimbriae is useful for the development of a vaccine composed of the fimbrial virulence factor. F18 fimbriae are composed of the major subunit FedA and the minor subunits FedE and the adhesin FedF. In the present study monoclonal antibodies (mAbs) against FedA and FedF were produced. In addition to their diagnostic value, these mAbs revealed a weaker interaction between FedA and FedF compared to the subunit-subunit interactions in other fimbriae, like type 1 and P pili. Further experiments are needed to investigate if this weak interaction could be one of the reasons for the slow colonisation of the small intestinal mucosa by F18+ E. coli. PMID- 17084565 TI - Etiology of respiratory disease in non-vaccinated, non-medicated calves in rearing herds. AB - The aim of this study was to examine the occurrence of bacterial, mycoplasmal and viral pathogens in the lower respiratory tract of calves in all-in all-out calf rearing units. According to clinical status, non-medicated calves with and without respiratory disease signs were selected of the 40 herds investigated to analyse the micro-organisms present in healthy and diseased calves. Tracheobronchial lavage (TBL) and paired serum samples were analysed for bacteria, mycoplasmas, respiratory syncytial virus (RSV), parainfluenza virus 3 (PIV3), bovine corona virus (BCV) and bovine adenovirus (BAV). Pasteurella multocida was the most common bacterial pathogen. It was isolated from 34% of the TBL samples in 28 herds and was associated with clinical respiratory disease (p < 0.05) when other pathogenic bacteria or mycoplasma were present in the sample. Mannheimia spp. and Histophilus somni were rarely found. Mycoplasma bovis was not detected at all. Ureaplasma diversum was associated with clinical respiratory disease (p < 0.05). TBL samples from healthy or suspect calves were more often negative in bacterial culture than samples from diseased calves (p < 0.05). No viral infections were detected in six herds, while 16-21 herds had RSV, BCV, BAV or PIV3. In the herds that had calves seroconverted to BCV, respiratory shedding of BCV was more frequently observed than faecal shedding. This study showed that the microbial combinations behind BRD were diverse between herds. M. bovis, an emerging pathogen in many countries, was not detected. PMID- 17084566 TI - Effects of soy protein and resistance exercise on body composition and blood lipids in postmenopausal women. AB - OBJECTIVE: To assess the effect of soy protein and progressive resistance training on body composition and lipids in postmenopausal women. DESIGN: In a controlled trial, 46 postmenopausal women were randomized to one of four groups: 25 g of soy protein (SP, n=10), 25 g of soy protein plus resistance exercise (SPE, n=14), 25 g of maltodextrine (placebo) (PL, n=11), or placebo plus resistance exercise (PLE, n=11). Progressive resistance training was held three times a week for 16 weeks and included 8 exercises (3 series of 8-12 repetitions). At baseline and after 16 weeks, body mass index, waist circumference (WC), body fat, muscle mass and serum lipid levels were measured. To confirm isoflavone absorption, urinary concentrations were determined. The t test of Student and ANOVA were used in the statistical analysis. RESULTS: Subjects were classified as overweight and showed android fat distribution. Urinary isoflavone excretion indicated compliance to soy protein treatment. After 16 weeks of intervention, both SPE and PLE groups showed a significant increase of 1.3 kg in muscle mass and reduction in WC of -1.4 and -2.1cm, respectively (p<0.05). Significant decreases in the mean values of total cholesterol and LDL ( 29.0 and -24.0 mg/dL, p<0.001 and p<0.006, respectively) were observed in the users of soy protein alone (SP). CONCLUSIONS: Soy protein supplementation did not influence the indicators of body composition. However, it exerted possible favorable effects on lipid profile in postmenopausal women. The increase in muscle mass and reduction in abdominal fat were correlated with resistance training. PMID- 17084567 TI - Dry powder aerosol delivery of large hollow nanoparticulate aggregates as prospective carriers of nanoparticulate drugs: effects of phospholipids. AB - The present work details the effects of incorporating phospholipids, a major component of lung surfactants, in the formulation of large hollow nanoparticulate aggregates, which are specifically designed to serve as potential carrier particles in inhaled delivery of nanoparticulate drugs. The large hollow aerosol particles (d(g) approximately 10 microm), whose shells are composed of nanoparticulate aggregates, are manufactured via the spray drying of nanoparticulate suspensions under a predetermined operating condition. Polyacrylate and silica nanoparticles of various sizes (20-170 nm), without loaded drugs, are employed as the model nanoparticles. The effects of increasing the phospholipids concentration in the presence of the nanoparticles, and vice versa, on the degree of hollowness and morphology of the spray-dried particles are investigated. Varying the phospholipids concentration in the presence of a constant amount of nanoparticles is found to influence the degree of hollowness, without significantly affecting the particle size distribution and respirable fine particle fraction, of the aerosol particles. The effects of increasing the phospholipids concentration on the degree of hollowness of the spray-dried particles are found to depend on the size and chemical nature of the nanoparticles. PMID- 17084568 TI - Anti-inflammatory and anti-allergic properties of the essential oil and active compounds from Cordia verbenacea. AB - The anti-inflammatory and anti-allergic effects of the essential oil of Cordia verbenacea (Boraginaceae) and some of its active compounds were evaluated. Systemic treatment with the essential oil of Cordia verbenacea (300-600mg/kg, p.o.) reduced carrageenan-induced rat paw oedema, myeloperoxidase activity and the mouse oedema elicited by carrageenan, bradykinin, substance P, histamine and platelet-activating factor. It also prevented carrageenan-evoked exudation and the neutrophil influx to the rat pleura and the neutrophil migration into carrageenan-stimulated mouse air pouches. Moreover, Cordia verbenacea oil inhibited the oedema caused by Apis mellifera venom or ovalbumin in sensitized rats and ovalbumin-evoked allergic pleurisy. The essential oil significantly decreased TNFalpha, without affecting IL-1beta production, in carrageenan injected rat paws. Neither the PGE(2) formation after intrapleural injection of carrageenan nor the COX-1 or COX-2 activities in vitro were affected by the essential oil. Of high interest, the paw edema induced by carrageenan in mice was markedly inhibited by both sesquiterpenic compounds obtained from the essential oil: alpha-humulene and trans-caryophyllene (50mg/kg, p.o.). Collectively, the present results showed marked anti-inflammatory effects for the essential oil of Cordia verbenacea and some active compounds, probably by interfering with TNFalpha production. Cordia verbenacea essential oil or its constituents might represent new therapeutic options for the treatment of inflammatory diseases. PMID- 17084569 TI - Volumetric evaluation of the paranasal sinuses in normal subjects using computer tomography images: a stereological study. AB - OBJECTIVE: The size and shape of paranasal sinuses are especially relevant when considering endoscopic sinus surgery. For this reason, the size of the paranasal sinuses has been the subject of many studies, none of which has used stereological methods to estimate the volume. In the present stereological study, we estimated the volume of paranasal sinuses of normal males and females. MATERIAL AND METHODS: We used a combination of the Cavalieri principle and computer tomography scans taken from 39 male and 38 female patients to estimate the volumes of frontal, maxillary, ethmoidal and sphenoidal sinuses. RESULTS: The mean volumes of frontal, maxillary, ethmoidal and sphenoidal sinuses were estimated bilaterally, producing mean volumes of 11.6+/-0.8, 35.9+/-1.3, 11.8+/ 0.4 and 13.6+/-0.7 cm(3), respectively. When the correlations between estimated volumes were analyzed statistically a positive relation was found for the paranasal sinuses. The size of the sinuses tends to decrease with age. CONCLUSION: The findings of the present study using the stereological methods could provide data for the evaluation of normal and pathological volumes of the paranasal sinuses. PMID- 17084570 TI - Low significance of MECP2 mutations as a cause of mental retardation in Brazilian males. AB - MeCP2 is a protein that selectively binds to methylated cytosines through its methyl-CpG-binding domain (MBD) and connects DNA methylation to transcriptional repression. Mutations in MECP2 gene, located in Xq28, have been reported as being the major cause of Rett syndrome and are also associated with some cases of X linked mental retardation in both males and females. In this study, we present the first screening in the MECP2 gene in a Brazilian cohort of 239 unrelated males with idiopathic mental retardation. Eight sequence variations were observed in 10 patients: one novel putative pathogenic variant, two never described variants of unknown pathogenic value and five non-pathogenic variations. We conclude that in mentally retarded Brazilian males, non-pathogenic variants in the MECP2 gene are more common than actual pathogenic mutations, and therefore alterations in this gene have a weak relationship with mental retardation in males. PMID- 17084571 TI - [Critical evaluation and predictive value of clinical presentation in out patients with acute community-acquired pneumonia]. AB - Diagnostic probability of community-acquired pneumonia (CAP) depends on data related to age and clinical and radiological findings. The critical evaluation of data in the literature leads to the following conclusions: 1) the prevalence of CAP in a given population with acute respiratory disease is 5% in outpatients and 10% in an emergency care unit. This could be as low as 2% in young people and even higher than 40% in hospitalized elderly patients; 2) the collection of clinical data is linked to the way the patient is examined and to the expertise of the clinician. The absolute lack of "vital signs" has a good negative predictive value in CAP; presence of unilateral crackles has a good positive predictive value; 3) there is a wide range of X-ray abnormalities: localized alveolar opacities; interstitial opacities, limited of diffused. The greatest radiological difficulties are encountered in old people with disorders including chronic respiratory or cardiac opacities and as a consequence of the high prevalence of bronchopneumonia episodes at this age; 4) among patients with lower respiratory tract (LRT) infections, the blood levels of leukocytes, CRP and procalcitonine are higher in CAP patients, mainly when their disease has a bacterial origin. Since you have not a threshold value reliably demonstrated in large populations with LRT infections or acute respiratory disease, presence or absence of these parameters could only be taken as a slight hint for a CAP diagnosis. PMID- 17084573 TI - Aging affects executive functions and memory in mouse lemur primates. AB - The decline of cognitive capacities with age in mouse lemur primates (Microcebus murinus) was assessed. Eight young adults (2-4 years) and nine aged adults (7-11 years) were examined on tasks designed to measure executive functions, procedural and declarative memory. The mouse lemurs were tested on the go-no go successive discrimination task, set shifting tasks (including extra-dimensional shift and reversal discrimination) and a spatial rule-guided discrimination task. There were four major findings. First, the deficits observed were not global but only on specific tasks indicating that only specific cognitive abilities are impaired with aging. Second, there were variations among aged subjects suggesting different patterns of cognitive aging. Third, alterations in cognitive abilities with aging in mouse lemurs seemed to be comparable to those described in aged monkeys and humans. Indeed, executive functions and declarative memory were affected in subpopulations of aged subjects whereas procedural memory remained intact in all the tested aged subjects. Finally, two forms of executive dysfunctions were distinguished among the aged subjects. The ultimate goal is to correlate age-related cognitive deficit with brain alterations and this study has helped to select candidate regions to be thoroughly scrutinized in aged mouse lemurs. PMID- 17084574 TI - Cigarette smoke--an aging accelerator? AB - Cigarette smoking reduces life span by an average of 7 years, and tobacco consumption accounts for a shortening of disease free life by 14 years. The exact mechanisms by which smoking causes disease and death are generally not well understood, but evidence continues to mount that cigarette smoking exhausts cellular defense and repair functions, leading to an accumulation of damage e.g. mutations and malfunctioning proteins. In this review, we make an attempt to ascribe many of the deleterious effects of smoking on human health to a general principle, namely the acceleration of aging processes by cigarette smoke chemicals. PMID- 17084575 TI - Stability of artesunate in pharmaceutical solvents. AB - Stability of artesunate (ART) was established in three pharmaceutical solvents. The chromatographic conditions developed for this study were acetonitrile:potassium phosphate buffer 10 mM (40:60, v:v; pH 2.9) at 0.7 mL min( 1) with UV detection at 220 nm using a short X-Terra RP C18 column (50 mm x 3 mm, 3.5 microm). This isocratic condition led to the separation between ART and its main degradation products (i.e. alpha-DHA and beta-DHA) with analysis time of less than 4 min. The retention factors are 1.49, 2.26 and 2.79 min for alpha-DHA, beta-DHA and ART, respectively. This method was proved linear (r(2)=0.9995), accurate (R.S.D.=0.20), precise (R.S.D.=0.74) and robust. The system performance remained unaffected by pH variation from 2.6 to 3.2 and variation of acetonitrile percentage from 38 to 42. Stability of ART was assessed in ethanol, propylene glycol (PG) and polyethylene glycol 400 (PEG 400). Unfortunately none of these solvents prevented ART from degradation longer than 3 months. In ethanol, significant degradation of ART occurred after 3 months at room temperature and this degradation was characterised by numerous degradation products. In PEG 400, significant degradation was observed after only 1 month, however DHA was the unique degradation product, which is also an efficient anti-malarial drug. PMID- 17084576 TI - Determination of oxcarbazepine by Square Wave Adsorptive Stripping Voltammetry in pharmaceutical preparations. AB - A procedure for the determination of oxcarbazepine (OXC) by Square Wave Adsorptive Stripping Voltammetry (SWAdSV) has been optimized. Selection of the experimental parameters was made using experimental design methodology. The detection limit was 1.74 x 10(-7) mol dm(-3). This method was used to determine oxcarbazepine in pharmaceutical preparations. PMID- 17084577 TI - Simultaneous determination of eight components in Radix Tinosporae by high performance liquid chromatography coupled with diode array detector and electrospray tandem mass spectrometry. AB - High-performance liquid chromatography (HPLC) coupled with electrospray tandem mass spectrometry (ESI-MS-MS) and diode array detection (DAD) was used to identify and simultaneously determine eight major ingredients in Radix Tinosporae. The assay was performed on a Diamonsil C(18) analytical column with a gradient solvent system of A (water containing 0.2% formic acid, 20mM ammonium acetate) and B (methanol/acetonitrile=1/1, v/v). The 217, 248, 270 and 347 nm, respectively, were chosen as the monitoring wavelengths to determine four structural types of components, say columbin, phytoecdysteroids (including 20 hydroxyecdysone, 2-deoxy-20-hydroxyecdysone 3-O-beta-d-glucopyranoside and 2 deoxy-20-hydroxyecdysone), menisperine and protoberberine alkaloids (including columbamine, jatrorrhizine and palmatine). This method was validated in respect to precision, repeatability and accuracy, and was successfully applied to quantify the eight components in 39 batches of R. Tinosporae for quality control purpose. The results indicated that the proposed method could be readily utilized as a quality control method for traditional Chinese medicine (TCM). PMID- 17084578 TI - Field cancerization in oral lichen planus. AB - BACKGROUND: The concept of field cancerization describes the tendency of patients with premalignant and malignant lesions of head and neck mucosal sites to develop multiple carcinomas of the upper aerodigestive tract. Here we address whether this concept should be extended also to patients affected by oral lichen planus (OLP), an inflammatory disorder associated with an increased risk of cancer development. METHODS: Data from a cohort of 45 patients with OLP who subsequently developed severe dysplastic changes and/or oral squamous cell carcinoma were retrospectively reviewed. Patients who presented more than one oral neoplastic event were considered for further data analysis as regards incidence, localization, management and prognosis. RESULTS: Twenty (44.4%) patients were affected by one single neoplastic event while 25 (55.6%) developed multiple and often multifocal oral dysplastic and/or malignant events. In most cases, a careful surveillance programme led to diagnosis and effective treatment of oral neoplasias at an early intraepithelial and microinvasive stage, leading to long term survival. In some patients, however, additional primary tumours occurred suddenly with rapid invasion, leading to advanced stage diagnosis and poor prognosis. Overall, three patients (12%) died due to malignant oral disease. CONCLUSIONS: Patients with OLP and subsequent development of dysplasia/ oral squamous cell carcinoma are at risk of having multiple and multifocal neoplastic events of the oral cavity, a phenomenon which parallels the concept of field cancerization of traditional head and neck cancers. If detected at an early stage, these neoplasias can be managed with superficial and complete resection. However a small number of patients have loco-regional tumour spread despite a standard surveillance protocol. PMID- 17084579 TI - Assessment of clinical palpation of the axilla as a criterion for performing the sentinel node procedure in breast cancer. AB - AIMS: Clinically palpable lymph nodes (LNs) are regarded as a contraindication for performing the sentinel node (SN) procedure. Many studies have shown, however, that clinical assessment of axillary LNs is inaccurate. This study evaluated the reliability of clinical axillary LN assessment by experts and assessed whether inaccuracy can be related to LN size. METHODS: Three hundred and one consecutive breast cancer patients undergoing either axillary dissection or SN were studied prospectively. RESULTS: The risk of having metastasis to the LN was 40.4% if the preoperative clinical assessment was "non-palpable LN", 61.5% if the assessment was "palpable but benign LN" and 84.4% if it was "suspicious LN". There were no clinically significant differences in mean size either when the LN was palpable versus non-palpable, or when the LN had metastasis or not. CONCLUSIONS: The clinical assessment of axillary LNs as a criterion for offering the SN procedure is of little value. PMID- 17084580 TI - Techniques for the placement of hepatic artery catheters for regional chemotherapy in unresectable liver metastases. AB - AIMS: For patients with unresectable liver metastases from colorectal cancer (CRC), hepatic artery infusion chemotherapy (HAIC) can produce higher response rates as compared to systemic chemotherapy. However, the added morbidity and early dysfunction after catheter placement have negatively influenced the outcomes of randomized studies. METHODS: Between July 1998 and March 2006, 29 patients with unresectable liver metastases from CRC were included in three different HAIC protocols. A catheter was laparoscopically placed in the gastroduodenal artery, retrograde to the common hepatic artery and was attached to a subcutaneous access port. Perioperative parameters and catheter-associated adverse events during chemotherapy were studied. RESULTS: Mean operating time was 106+/-29 min and median duration of hospitalization was 2 days (range: 1-13). No major perioperative complications occurred but there was 1 death (3.6%) 1 week after surgery due to hemorrhagic shock after necrosis and rupture of the hepatic artery. During a median follow-up of 10 months (range: 0-35) we observed 5 (17%) catheter-associated adverse events, but in only 2 patients (7%) this resulted in a loss of catheter function before the end of chemotherapy. Overall median duration of HAIC was 8 months (range: 1-28). CONCLUSION: Laparoscopic placement of a hepatic artery catheter is associated with a low operative morbidity. The observed functionality of the hepatic artery ports in our series is encouraging for the use in future studies of HAIC. PMID- 17084581 TI - Neither the presence of ica locus, nor in vitro-biofilm formation ability is a crucial parameter for some Staphylococcus epidermidis strains to maintain an infection in a guinea pig tissue cage model. AB - The pathogenesis of Staphylococcus epidermidis is thought to be based on its capacity to colonize medical devices by forming a biofilm. Biofilm formation is in part mediated by the polysaccharide intercellular adhesin (PIA), which is encoded by the icaADBC operon. We have previously investigated in vitro the correlation existing between biofilm formation (B+/-), presence of ica locus (I+/ ) and PIA production (P+/-) in some clinical isolates of coagulase-negative staphylococci (CoNS). Here, we used a guinea pig model of subcutaneous implanted tissue cages to assess the implication of B, I and P parameters in the capacity of nine S. epidermidis and one S. carnosus strains to develop and maintain an infection in vivo. Using clinical isolates and a model strain of S. epidermidis, we showed that the "B+, I+, P+" type confers the ability to maintain an infection in vivo. Surprisingly, the opposite type "B-, I-, P-" tested with clinical and commensal isolates, presented infection rates ranging from 25% to 60%. Other clinical isolates having a "B+, I+, P-" type, were not able to cause an infection in the present model. These results showed that, depending on the strains the capacity to colonize the tissue cage might be independent of the ability to form biofilm. PMID- 17084582 TI - In vitro models for the study of the effect of butyrate on human colon adenocarcinoma cells. AB - The effect of butyrate has been analyzed on human colon adenocarcinoma cell lines with different properties regarding tumorigenicity, differentiation and resistance to apoptosis induced by this agent. Butyrate reduces cell proliferation, induces differentiation (according to alkaline phosphatase activity) and apoptosis, being these effects time- and concentration-dependent. The susceptibility to the cytotoxic effects of butyrate depends on the cell line considered and it is not directly related to tumorigenicity or differentiation. We show that 2mM butyrate treatment of non-tumorigenic BCS-TC2 cells for four days strongly influences the transcriptional activity, causing extensive modification in gene expression patterns (69 up-regulated and 109 down-regulated genes). Some of these genes are involved in the modulation of cell cycle progression, apoptosis and differentiation. We have analyzed the effect of butyrate in spontaneous or induced multicellular spheroids. The more stable spheroids (spontaneous or induced from butyrate-resistant cells) increase the resistance of cells to the effects of butyrate probably due to an impaired accessibility. This in vitro model could be useful to study the resistance of tumors to the effect of natural regulators (i.e. butyrate) as well as to develop and test new therapeutic approaches. PMID- 17084583 TI - Induction of apoptosis in Jurkat cells by photoexcited psoralen derivatives: Implication of mitochondrial dysfunctions and caspases activation. AB - The prevailing form of cell death in lymphocytes exposed to psoralen plus UVA light (PUVA), was investigated. We studied the well known drug 8-methoxypsoralen (8-MOP) and an angular derivatives: angelicin (ANG). We evaluated the induction of apoptosis in a human tumor T-cell line (Jurkat). Both compounds provoke a significant induction of apoptosis at 24h from irradiation as demonstrated by a remarkable percentage of cells Annexin-V positive. We investigated the effects of the psoralen derivatives upon UVA irradiation on the cell cycle. The flow cytometric analysis of propidium labeled cells indicates that treatment induces, in a dose dependent manner, a massive accumulation of cells, for both compounds, in G2-S phase after 24h from the irradiation. We have focused our attention on the mitochondrial functionality after irradiation in the presence of psoralen derivatives. We evaluated, by flow cytometry, (i) the mitochondrial potential (Deltapsi(mt)), (ii) the production of reactive oxygen species (ROS) and (iii) the oxidation of cardiolipin, a phospholipid restricted to the inner mitochondrial membrane. Furthermore the activation of caspases -3, -8 and -9 was also investigated. The obtained data indicated that, upon UVA irradiation, the two compounds induce a strong decrease in mitochondrial functions and activate caspase-3, -8 and -9. PMID- 17084584 TI - Effect of different plant extracts and natural substances (PENS) against membrane damage induced by enterotoxigenic Escherichia coli K88 in pig intestinal cells. AB - Pig weaning period is frequently associated with infectious disease, mainly caused by enterotoxigenic Escherichia coli (ETEC) K88. Plant extracts exert different beneficial effects and may represent antibiotic alternatives to reduce piglet infection. In this study, plant extracts and other natural substances (PENS) have been evaluated on the pig intestinal IPEC-1 cells, for potential protection against ETEC K88 induced membrane damage. Several PENS have been considered: yeast extract, yeast nucleotides, unsaturated oligo-mannuronic acid, ulvan, bromelain and three fractions of bovine colostrums, as anti-inflammatory and immunomodulatory compounds; daidzein and Chlorella vulgaris extract, as anti oxidant compounds; allicin, cinnamaldehyde and carvacrol, as anti-bacterial compounds. First, possible toxic effect of PENS on cell membrane permeability was verified by assessing the transepithelial electrical resistance (TEER) and paracellular flux of the extracellular marker phenol red. The highest non-toxic PENS concentration was added to ETEC infected cells to test the protection against membrane damage. The results showed that yeast extract, daidzein, bovine colostrum, bromelain and allicin protected the cells against the increased membrane permeability caused by ETEC, whereas the other PENS did not show this ability. Allicin protection was not due to its anti-bacterial activity, since ETEC growth was unaffected by the presence of allicin. PMID- 17084585 TI - Antioxidative and oxidative changes in the digestive gland cells of freshwater mussels Unio tumidus caused by selected phenolic compounds in the presence of H(2)O(2) or Cu(2+) ions. AB - Research on biomarkers as early bioindicators of perturbation in populations and individuals has received increasing interest during recent decades. These ecotoxicity studies allow us to measure the impact of environmental stressors and to monitor and evaluate the degradation or restoration of systems. In the present study we used bivalve molluscs (mussels), which are sensitive biomarkers of aquatic ecosystem pollution, to assess the effects of three polyphenols: tannic acid, ellagic acid and gallic acid. These compounds were used in the 1-60 microM concentration range, alone and in the presence of H(2)O(2) (40 and 100 microM) or Cu(2+) ions (50 microM). The fluorescence probe dichlorofluorescein-diacetate (DCFH-DA) was used to measure reactive oxygen species (ROS). The oxidation of DCFH-DA to the fluorescent DCF (dichlorofluorescein) by the phenolic compounds was investigated spectrofluorimetrically. The results showed that the polyphenols tested can act as antioxidants against the ROS present in the digestive gland cells, but their activity is decreased after incubation with hydrogen peroxide or copper ions. SH-groups were determined spectrophotometrically using Ellman's reagent. The results showed that oxidative modification of proteins increased in a concentration-dependent manner in cells incubated with polyphenols (above 15 microM) alone. Incubation of the cells with phenolic acids and H(2)O(2) or Cu(2+) ions revealed that the phenolic acids had prooxidant properties in all concentrations used except for 1 microM tannic and ellagic acid and 40 microM H(2)O(2). DNA fragmentation was estimated by a fluorescence method using Hoechst 33258/propidium iodine binding. The data showed that the phenolic acids alone and in the presence of hydrogen peroxide or copper ions can induce apoptosis and necrosis. The methods used and results obtained indicate that the polyphenols selected can act not only as antioxidants but also as prooxidants in digestive gland cells of Unio tumidus. PMID- 17084586 TI - Interleukin-1beta expression in murine J774A.1 macrophages exposed to platinum compounds: the role of p38 and ERK 1/2 mitogen-activated protein kinases. AB - Although skin and respiratory sensitizing properties of platinum compounds have been proved in humans and mice, little is known about signal transduction pathways leading to cytokine production in the induction phase. It is generally assumed that induction of skin sensitization, but not skin irritation, is associated with a rapid increase in the IL-1beta mRNA expression. In this study, IL-1beta expression and a role of mitogen-activated protein kinases (MAPKs) in this process were investigated in murine macrophages J774A.1 exposed to four platinum compounds. Potassium tetrachloroplatinate (K(2)PtCl(4); TCPP), ammonium tetrachloroplatinate ((NH(4))(2)PtCl(4); TCPA), ammonium hexachloroplatinate ((NH(4))(2)PtCl(6); HCPA) showed a very similar range of cytotoxic concentrations (IC(50) values: 238 microM+/-30; 269 microM+/-39 and 245 microM+/-31, respectively) as assessed in the 24-h MTT reduction test. Cytotoxicity of cis diammineplatinum dichloride (cisplatin) was considerably higher (IC(50) of 23 microM+/-4). While increased expression of IL-1beta mRNA was observed in the macrophages exposed to each test compound, IL-1beta protein production was detected in cell lysates after treatment with TCPP, TCPA and HCPA for 24h (concentration range of 150-350 microM) as well as for 2h (450-650 microM). The treatment with each compound resulted in the phosphorylation of both p38 MAPK and ERK 1/2 (p44/42). Blocking the activation of p38 MAPK as well as ERK 1/2 with specific inhibitors (SB203580 and U0126, respectively) down-regulated the IL 1beta expression. Interestingly, the skin irritant sodium dodecyl sulfate did not trigger phosphorylation of these kinases, nor induced IL-1beta production. These data suggest that p38 MAPK and ERK 1/2 play an important role in induction of IL 1beta expression in J774A.1 macrophages exposed to test platinum compounds. PMID- 17084587 TI - Acquisition of resistance to butyrate induces resistance to luminal components and other types of stress in human colon adenocarcinoma cells. AB - Butyrate, naturally produced by anaerobic fermentation of diet-fiber, is the major nutrient of colonocytes and also an important regulator of colonic epithelium renewal and physiology. Other luminal components, such as bile acids or bacterial products, influence these processes. The model system we used to analyze the influence of several luminal stressors is composed of a previously established cell line resistant to the apoptotic effects of butyrate and their parental butyrate-sensitive cells. Viability of butyrate-resistant cells is unaffected by mild heat-shock (2h, 42 degrees C) and only slightly reduced by severe heat-shock (2h, 45 degrees C) in contrast to their butyrate-sensitive counterparts. The higher constitutive expression of HSP70 and HSP60 could contribute to this resistance. In addition, expression of HSP70 follows a different pattern after heat-shock in both cell lines. Butyrate-resistant cells are quite unaffected by treatment with deoxycholic acid but apoptosis is induced by chenodeoxycholic acid although to a lower extent than in butyrate-sensitive cells. These resistant cells are also less sensitive to lipopolysaccharide and show differences regarding the activation of ERK following osmotic stress. Thus, the cell model herein reported is a useful tool for investigating the molecular mechanisms of resistance to apoptosis, as well as to analyze specific targets for butyrate-resistant tumors. PMID- 17084588 TI - Screening of potentially toxic chalcogens in erythrocytes. AB - Previous literature reports have demonstrated that a number of human diseases, including inflammation and cancer, can be caused by environmental and occupational exposure to toxic compounds, via DNA damage, protein modifications, or lipid peroxidation. The present study was undertaken to screen the toxicity of a variety of chalcogens using erythrocytes as a model of cell injury. The toxicity of these compounds was evaluated via quantification of hemolysis and lipid peroxidation. The present investigation shows that diphenyl ditelluride and phenyl tellurides are toxic to erythrocytes. The organoselenium compounds were not toxic to erythrocytes even when tested at high concentrations and with a hematocrit of 45%. The hemolytic effect of tellurides was not positively correlated with thiobarbituric acid-reactive substance (TBARS) production suggesting that lipid peroxidation is not involved in the hemolysis provoked by organotellurium compounds. The results suggest that chalcogen compounds may be toxic to human erythrocytes, depending on their structure. PMID- 17084589 TI - Psychometric properties of the Social Interaction Anxiety Scale and separation criterion between Spanish youths with and without subtypes of social anxiety. AB - In this study, we analyzed the reliability and validity of the Social Interaction Anxiety Scale (SIAS) and propose a separation criterion between youths with specific and generalized social anxiety and youths without social anxiety. A sample of 1012 Spanish youths attending school completed the SIAS, the Liebowitz Social Anxiety Scale, the Social Avoidance and Distress Scale, the Fear of Negative Evaluation Scale, the Youth Self-Report for Ages 11-18 and the Minnesota Multiphasic Personality Inventory-Adolescent. The factor analysis suggests the existence of three factors in the SIAS, the first two of which explain most of the variance of the construct assessed. Internal consistency is adequate in the first two factors. The SIAS features an adequate theoretical validity with the scores of different variables related to social interaction. Analysis of the criterion scores yields three groups pertaining to three clearly differentiated clusters. In the third cluster, two of social anxiety groups - specific and generalized - have been identified by means of a quantitative separation criterion. PMID- 17084590 TI - Optimized linkage and quenching strategies for quantum dot molecular beacons. AB - Quantum dot (QD) molecular beacons were explored for sequence-specific DNA detection. The effectiveness of multiple linkage strategies and fluorescence quenchers were compared in hybridization-based assays. To compare linkage strategies, covalent amide linkage and streptavidin-biotin binding were used to link semiconductor QDs to molecular beacon DNA. Amide-linked beacons showed a 57% greater fluorescence increase than streptavidin-linked beacons when hybridized to 200 pmol of target DNA. The specificity of the molecular beacons, however, was similar for both linkage methods. Hybridization of both QD molecular beacons with non-complementary target DNA resulted in approximately 50% lower fluorescence intensity than hybridization with complementary DNA. The effectiveness of different quencher moieties was also evaluated. Iowa Black and 1.4 nm Nanogold quenched molecular beacons exhibited approximately 2-fold greater fluorescence increases than dabcyl-quenched beacons when hybridized to complementary target. Specificity for target DNA was also confirmed through hybridization assays with non-complementary DNA. To provide insight into differences between the QD molecular beacons and the linkage strategies used, the hydrodynamic radius of each was measured. These measurements indicated that the larger radius of the streptavidin QDs (13.5 nm) than the carboxyl QDs (7 nm) could have a negative effect on FRET-based quenching for QD molecular beacons. These data outline the importance of choosing proper linkage methods and quencher moieties for creating high-quality QD molecular beacons. PMID- 17084591 TI - The effects of frequency range, vowel, dynamic loudness level, and gender on nasalance in amateur and classically trained singers. AB - This study addresses two questions: (1) How much nasality is present in classical Western singing? (2) What are the effects of frequency range, vowel, dynamic level, and gender on nasality in amateur and classically trained singers? The Nasometer II 6400 by KayPENTAX (Lincoln Park, NJ) was used to obtain nasalance values from 21 amateur singers and 25 classically trained singers while singing an ascending five-tone scalar passage in low, mid, and high frequency ranges. Each subject sang the scalar passage at both piano and mezzo-forte dynamic loudness levels on each of the five cardinal vowels (/a/, /e/, /i/, /o/, /u/). A repeated mixed-model analysis indicated a significant main effect for the amateur/classically trained distinction, dynamic loudness level, and vowel, but not for frequency range or gender. The amateur singers had significantly higher nasalance scores than classically trained singers in all ranges and on all vowels except /o/. Dynamic loudness level had a significant effect on nasalance for all subject groups except for female majors in the mid- and high-frequency ranges. The vowel, /i/, received significantly higher nasalance than all of the other vowels. Although results of this study show that dynamic loudness level, vowel, and level of training in classical singing have a significant effect on nasality, nasalance scores for most subjects were relatively low. Only six of the subjects, all of whom were amateur singers, had average nasalance scores that could be considered hypernasal (ie, a nasalance average of 22 or above). PMID- 17084593 TI - Antidiabetic effect of probiotic dahi containing Lactobacillus acidophilus and Lactobacillus casei in high fructose fed rats. AB - OBJECTIVE: We investigated the effect of low-fat (2.5%) dahi containing probiotic Lactobacillus acidophilus and Lactobacillus casei on progression of high fructose induced type 2 diabetes in rats. METHODS: Diabetes was induced in male albino Wistar rats by feeding 21% fructose in water. The body weight, food and water intakes, fasting blood glucose, glycosylated hemoglobin, oral glucose tolerance test, plasma insulin, liver glycogen content, and blood lipid profile were recorded. The oxidative status in terms of thiobarbituric acid-reactive substances and reduced glutathione contents in liver and pancreatic tissues were also measured. RESULTS: Values for blood glucose, glycosylated hemoglobin, glucose intolerance, plasma insulin, liver glycogen, plasma total cholesterol, triacylglycerol, low-density lipoprotein cholesterol, very low-density lipoprotein cholesterol, and blood free fatty acids were increased significantly after 8 wk of high fructose feeding; however, the dahi-supplemented diet restricted the elevation of these parameters in comparison with the high fructose fed control group. In contrast, high-density lipoprotein cholesterol decreased slightly and was retained in the dahi-fed group. The dahi-fed group also exhibited lower values of thiobarbituric acid-reactive substances and higher values of reduced glutathione in liver and pancreatic tissues compared with the high fructose-fed control group. CONCLUSION: The probiotic dahi-supplemented diet significantly delayed the onset of glucose intolerance, hyperglycemia, hyperinsulinemia, dyslipidemia, and oxidative stress in high fructose-induced diabetic rats, indicating a lower risk of diabetes and its complications. PMID- 17084592 TI - Cell biology of polycystin-2. AB - Naturally occurring mutations in two separate, but interacting loci, pkd1 and pkd2 are responsible for almost all cases of autosomal dominant polycystic kidney disease (ADPKD). ADPKD is one of the most common genetic diseases resulting primarily in the formation of large kidney, liver, and pancreatic cysts. Homozygous deletion of either pkd1 or pkd2 results in embryonic lethality in mice due to kidney and heart defects illustrating their indispensable roles in mammalian development. However, the mechanism by which mutations in these genes cause ADPKD and other developmental defects are unknown. Research in the past several years has revealed that PKD2 has multiple functions depending on its subcellular localization. It forms a receptor-operated, non-selective cation channel in the plasma membrane, a novel intracellular Ca2+ release channel in the endoplasmic reticulum (ER), and a mechanosensitive channel in the primary cilium. This review focuses on the functional compartmentalization of PKD2, its modes of activation, and PKD2-mediated signal transduction. PMID- 17084594 TI - Dietary pectin stimulates protein metabolism in the digestive tract. AB - OBJECTIVE: The aim of this study was to determine if protein metabolism was altered in small and large intestines by feeding pectin, a soluble fiber known to stimulate cecal production of short-chain fatty acids (SCFAs) and to have a trophic effect in these tissues. METHODS: Twenty-four weanling male Sprague Dawley rats were fed ad libitum for 14 d with a balanced control diet or an isoproteic, isocaloric pectin (citrus) diet (80 g/kg). SCFA production, intestinal histomorphometry, and protein synthesis were determined in the proximal and distal parts of the small intestine, the cecum, and the colon. Protein synthesis rates were determined by measuring the (13)C valine incorporation rate in tissue proteins. RESULTS: Pectin feeding slightly decreased food intake and growth rate. It increased the acetate, propionate, and butyrate pools in the cecum. Pectin feeding resulted in heavier intestinal tissues corresponding to higher villus height in the small intestine and crypt depth in the small and large intestines compared with feeding of the control diet. Compared with the control group, the rats fed the pectin diet had significantly higher protein synthesis rates in all the parts of their intestines. CONCLUSION: Supplementation of pectin, as a soluble fiber, in the diets, stimulated SCFA production, had a trophic effect on the different parts of the intestines, and greatly stimulated protein synthesis in those tissues. PMID- 17084595 TI - Intralipid minimizes hepatocytes injury after anoxia-reoxygenation in an ex vivo rat liver model. AB - OBJECTIVE: Ischemia-reperfusion injury is a determinant in liver injury occurring during surgical procedures, ischemic states, and multiple organ failure. The pre existing nutritional status of the liver, i.e., fasting, might contribute to the extent of tissue injury. This study investigated whether Intralipid, a solution containing soybean oil, egg phospholipids, and glycerol, could protect ex vivo perfused livers of fasting rats from anoxia-reoxygenation injury. METHODS: The portal vein was cannulated, and the liver was removed and perfused in a closed ex vivo system. Isolated livers were perfused with glucose 5.5 and 15 mM, and two different concentrations of Intralipid, i.e., 0.5:100 and 1:100 (v/v) Intralipid 10%:medium (n = 5 in each group). The experiment consisted of perfusion for 15 min, warm anoxia for 60 min, and reoxygenation during 60 min. Hepatic enzymes, potassium, glucose, lactate, bilirubin, dienes, trienes, and cytochrome-c were analyzed in perfusate samples. The proportion of glycogen in hepatocytes was determined in biopsies. RESULTS: Intralipid attenuated transaminases, lactate dehydrogenase, potassium, diene, and triene release in the perfusate (dose dependant) during the reoxygenation phase when compared with glucose-treated groups. The concentration of cytochrome-c in the medium was the highest in the 5.5-mM glucose group. The glycogen content was low in all livers at the start of the experiment. CONCLUSION: Intralipid presents, under the present experimental conditions, a better protective effect than glucose in anoxia-reoxygenation injury of the rat liver. PMID- 17084596 TI - Violence and schizophrenia. PMID- 17084597 TI - On the microstructures of ethyl acetate/monoolein/water. AB - A phase diagram, describing the behavior of the polar lipid monoolein (MO), water, and ethyl acetate (EtAc), is here presented as well as results from small angle X-ray scattering. MO is found to have a solubility of 60wt.% in EtAc at 20 degrees C. No macroscopic aggregation of MO can, initially, be detected in the binary MO/EtAc solution even though MO forms solid crystals in concentrated samples when times goes by. In case of the ternary system small amounts of water, mainly bound to the lipid head groups, can be incorporated in the liquid EtAc/MO phase as water has a limited solubility in EtAc. For EtAc/water mass ratios below 2/3 EtAc is present into the reversed bicontinuous cubic and lamellar phases present in the binary MO/water system. To conclude, EtAc is mainly partitioned to the lipid membranes, with minor effects on spontaneous curvature. Hence, simple EtAc-addition has an effect similar to dehydration. For EtAc/water ratios above 2/3 the liquid crystalline phases dissolve. The phase behavior is here discussed in view of related phase behaviors for water-miscible solvent/MO/water systems. For instance, an interpretation of the swelling behavior of the sponge phase (L(3)), present in the water-miscible solvent(s)/MO/water systems, shows that solvents partitioned to the polar domains strongly increases the spontaneous curvature of the MO-films. The reason is probably weaker hydrophobic interactions in interfacial regions. As expected, in case of water-miscible solvents, the ternary phase behaviors can be understood by consider water and water-miscible solvent as one "mixed solvent". PMID- 17084598 TI - Subcellular topological effect of particle monolayers on cell shapes and functions. AB - We studied topological effects of subcellular roughness displayed by a closely packed particle monolayer on adhesion and growth of endothelial cells. Poly(styrene-co-acrylamide) (SA) particles were prepared by soap-free emulsion copolymerization. Particle monolayers were prepared by Langmuir-Blodgett deposition using particles, which were 527 (SA053) and 1270 nm (SA127) in diameter. After 24-h incubation, cells tightly adhered on a tissue culture polystyrene dish and randomly spread. On the other hand, cells attached on particle monolayers were stretched into a narrow stalk-like shape. Lamellipodia spread from the leading edge of cells attached on SA053 monolayer to the top of the particles and gradually gathered to form clusters. This shows that cell-cell adhesion became stronger than cell-substrate interaction. Cells attached to SA127 monolayer extended to the reverse side of a particle monolayer and engulfed particles. They remained immobile without migration 24h after incubation. This shows that the inhibition of extensions on SA127 monolayer could inhibit cell migration and cell proliferation. Cell growth on the particle monolayers was suppressed compared with a flat TCPS dish. The number of cells on SA053 gradually increased, whereas that on SA127 decreased with time. When the cell seeding density was increased to 200,000 cells cm(-2), some adherent cells gradually became into contact with adjacent cells. F-actin condensations were formed at the frame of adherent cells and the thin filaments grew from the edges to connect each other with time. For the cell culture on SA053 monolayer, elongated cells showed a little alignment. Cells showed not arrangement of actin stress fibers but F-actin condensation at the contact regions with neighboring cells. Interestingly, the formed cell monolayer could be readily peeled from the particle monolayer. These results indicate that endothelial cells could recognize the surface roughness displayed by particle monolayers and the response was dependent on the pitch of particle monolayers. PMID- 17084599 TI - Estimation of synaptic conductances. AB - In order to identify and understand mechanistically the cortical circuitry of sensory information processing estimates are needed of synaptic input fields that drive neurons. From intracellular in vivo recordings one would like to estimate net synaptic conductance time courses for excitation and inhibition, g(E)(t) and g(I)(t), during time-varying stimulus presentations. However, the intrinsic conductance transients associated with neuronal spiking can confound such estimates, and thereby jeopardize functional interpretations. Here, using a conductance-based pyramidal neuron model we illustrate errors in estimates when the influence of spike-generating conductances are not reduced or avoided. A typical estimation procedure involves approximating the current-voltage relation at each time point during repeated stimuli. The repeated presentations are done in a few sets, each with a different steady bias current. From the trial-averaged smoothed membrane potential one estimates total membrane conductance and then dissects out estimates for g(E)(t) and g(I)(t). Simulations show that estimates obtained during phases without spikes are good but those obtained from phases with spiking should be viewed with skeptism. For the simulations, we consider two different synaptic input scenarios, each corresponding to computational network models of orientation tuning in visual cortex. One input scenario mimics a push pull arrangement for g(E)(t) and g(I)(t) and idealized as specified smooth time courses. The other is taken directly from a large-scale network simulation of stochastically spiking neurons in a slab of cortex with recurrent excitation and inhibition. For both, we show that spike-generating conductances cause serious errors in the estimates of g(E) and g(I). In some phases for the push-pull examples even the polarity of g(I) is mis-estimated, indicating significant increase when g(I) is actually decreased. Our primary message is to be cautious about forming interpretations based on estimates developed during spiking phases. PMID- 17084600 TI - The role of predictive models in the formation of auditory streams. AB - Sounds provide us with useful information about our environment which complements that provided by other senses, but also poses specific processing problems. How does the auditory system distentangle sounds from different sound sources? And what is it that allows intermittent sound events from the same source to be associated with each other? Here we review findings from a wide range of studies using the auditory streaming paradigm in order to formulate a unified account of the processes underlying auditory perceptual organization. We present new computational modelling results which replicate responses in primary auditory cortex [Fishman, Y.I., Arezzo, J.C., Steinschneider, M., 2004. Auditory stream segregation in monkey auditory cortex: effects of frequency separation, presentation rate, and tone duration. J. Acoust. Soc. Am. 116, 1656-1670; Fishman, Y. I., Reser, D. H., Arezzo, J.C., Steinschneider, M., 2001. Neural correlates of auditory stream segregation in primary auditory cortex of the awake monkey. Hear. Res. 151, 167-187] to tone sequences. We also present the results of a perceptual experiment which confirm the bi-stable nature of auditory streaming, and the proposal that the gradual build-up of streaming may be an artefact of averaging across many subjects [Pressnitzer, D., Hupe, J. M., 2006. Temporal dynamics of auditory and visual bi-stability reveal common principles of perceptual organization. Curr. Biol. 16(13), 1351-1357.]. Finally we argue that in order to account for all of the experimental findings, computational models of auditory stream segregation require four basic processing elements; segregation, predictive modelling, competition and adaptation, and that it is the formation of effective predictive models which allows the system to keep track of different sound sources in a complex auditory environment. PMID- 17084601 TI - Naturally occurring neuronal NO-synthase inactivators found in Nicotiana tabacum (Solanaceae) and other plants. AB - NO-synthase (NOS) is a heme-containing enzyme that catalyzes the oxidation of L arginine to nitric oxide, an important cellular signaling molecule. Recently, it was found that aqueous extracts of tobacco cigarettes cause the inactivation of the neuronal isoform of NOS (nNOS) and that this may explain some of the toxicological effects of smoking. Although the exact identity of the chemical inactivator(s) is not known, we wondered if extracts prepared from other plants, including those closely related to tobacco, Nicotiana tabacum (Solanaceae), would similarly inactivate nNOS. We examined 33 plants, representing diverse members of the plant kingdom ranging from whisk fern, Psilotum nudum (Psilotaceae) to tobacco and discovered 18 plants that contain a chemical inactivator(s) of nNOS. Of these plants, 16 are members of the core asterids flowering plant group. Of these asterids, 6 are members of the Solanaceae family, of which tobacco is a member. Based on the phylogenetic relationship of the plants, it is possible that the same chemical or related chemical inactivator(s) exist. This, in turn, may help elucidate the structure of the chemical(s), as well as provide a source of a potentially novel inactivator of nNOS. The alkaloid nicotine can be excluded as putative nNOS inhibitor. PMID- 17084602 TI - Immunomodulatory activity of Sulforaphane, a naturally occurring isothiocyanate from broccoli (Brassica oleracea). AB - The effect of Sulforaphane on the immune system was studied using BALB/c mice. Intraperitoneal administration of five doses of Sulforaphane (500 microg/dose/animal/day) was found to enhance the total WBC count (12,950 cells/mm3) on 9th day. Bone marrow cellularity (23 x 10(6) cells/femur) and number of alpha-esterase positive cells (1346.66/4000 cells) were also increased by the administration of Sulforaphane. Treatment with Sulforaphane along with the antigen, sheep red blood cells (SRBC), produced an enhancement in the circulating antibody titre and the number of plaque forming cells (PFC) in the spleen. Maximum number of PFC (315.83 PFC/10(6) spleen cells) was obtained on the 6th day. Administration of Sulforaphane also showed an enhancement in the phagocytic activity of peritoneal macrophages. Moreover administration of Sulforaphane significantly reduced the elevated level of TNF-alpha production by LPS stimulated macrophages. These results indicate the immunomodulatory activity of Sulforaphane. PMID- 17084603 TI - Inhibitors of aldose reductase and advanced glycation end-products formation from the leaves of Stelechocarpus cauliflorus R.E. Fr. AB - Two dihydroflavonol glycosides, engeletin and astilbin, were isolated from an EtOAc extract of the leaves of Stelechocarpus cauliflorus R.E. Fr. (Annonaceae). The inhibitory activity of engeletin against a recombinant human aldose reductase (IC50 value=1.16 microM) was twice that of quercetin as a positive control (2.48 microM), and 23 times greater than that of astilbin (26.7 microM). Engeletin inhibited the enzyme uncompetitively. Astilbin was about as potent as the positive control, quercetin, in its inhibition of advanced glycation end-products formation. These flavonoids displayed therapeutic potential in the prevention and treatment of diabetic complications. PMID- 17084604 TI - Beta-secretase (BACE1)-inhibiting stilbenoids from Smilax Rhizoma. AB - In the course of searching for BACE1 (beta-secretase) inhibitors from natural products, the ethyl acetate soluble fraction of Smilax Rhizoma (the dried rhizomes of Smilax china L.) showed potent inhibitory activity. The active compounds were identified as a trans/cis-resveratrol mixture, oxyresveratrol, veraphenol, and cis-scirpusin A. They were shown to non-competitively inhibit BACE1 with the Ki values of 5.4 x 10(-6), 5.4 x 10(-6), 3.4 x 10(-6), and 5.4 x 10(-6)M and IC(50) values of 1.5 x 10(-5), 7.6 x 10(-6), 4.2 x 10(-6), and 1.0 x 10(-5)M, respectively. The active compounds were less inhibitory to alpha secretase (TACE) and other serine proteases such as chymotrypsin, trypsin, and elastase, suggesting that they were relatively specific inhibitors of BACE1. PMID- 17084605 TI - Stimulating effects on mouse splenocytes of glycoproteins from the herbal medicine Atractylodes macrocephala Koidz. AB - A traditional herbal medicine, Atractylodes macrocephala Koidz. (AMK), has long been used as a digestive and tonic. Recent investigations have suggested its potential ability in stimulating immune responses, although a scientific basis for this activity has not yet been elucidated. Based on previous results showing that the activity might be due to proteins, we purified protein samples from an original sample preparation of AMK and examined the stimulating ability of the protein samples on mouse splenocytes. The sample treatment markedly stimulated lymphocyte proliferation, antibody production, and cytokine secretion in mouse splenocytes. In particular, the samples showed the ability to induce the preferential stimulation of Th1 type, rather than Th2 type T lymphocytes. Stimulating activity of the samples was associated closely with glycoprotein(s) with molecular weights of around 30 kDa, especially with carbohydrate moiety rather than with protein residues of the glycoprotein(s). Our findings suggest that the glycoprotein(s) might play critical roles in modulating immune-response induction, and could potentially be used as medicinal and pharmacological agents. PMID- 17084606 TI - Lipid peroxidation, cyclooxygenase enzyme and tumor cell proliferation inhibitory compounds in Cornus kousa fruits. AB - The genus Cornus is well known for its medicinal properties. Bioassay-guided isolation and characterization of C. kousa fruits afforded kaempferol 3-O rhamnoside (1), myricetin 3-O-rhamnoside (2), kaempferol 3-O-glucoside (3), cornin (4) and stenophyllin (5) in addition to ursolic acid and beta-sitosterol. These compounds are isolated for the first time from C. kousa. Compounds 1-5 inhibited Fe(2+) catalyzed lipid peroxidation by 63%, 57%, 61%, 53%, and 51%, at 23, 22, 23, 129, and 108 microM, respectively. Similarly, they inhibited COX-1 and -2 enzymes activities by 24% and 47%, 40% and 37%, 20% and 37%, 52% and 63%, and 48% and 55% respectively, at 231, 215, 226, 258, and 217 microM, respectively. At 129 microM, compound 4 displayed growth inhibition of HCT-116 (colon), MCF-7 (breast), NCI-H460 (lung), SF-268 (central nervous system CNS), and AGS (stomach) human tumor cell lines by 31%, 29%, 40%, 9%, and 28%, respectively. Similarly, compound 5 inhibited the growth of colon, breast, lung, CNS, and stomach tumor cell lines by 0%, 27%, 35%, 16%, and 27%, respectively, at 108 microM. PMID- 17084607 TI - Female gender is associated with higher incidence and more stable respiratory symptoms during adolescence. AB - Childhood asthma and wheeze is more common among boys than girls, while the opposite is found in adults. The main objective was to study the incidence and the course of wheeze and asthma during adolescence with focus on gender differences. In addition, we explored associations between lifestyle factors at baseline and wheeze at follow-up. A total of 2399 adolescents answered validated questionnaires on respiratory symptoms and lifestyle in 1995-1997 (13-15 years) and at follow-up in 2000-2001 (17-19 years). The risk of reporting wheeze and asthma at follow-up was greater in girls compared to boys among subjects reporting no respiratory symptoms at baseline; Relative risk: 1.4 and 2.4, respectively. More girls than boys reported current wheeze at follow-up, both among those with current wheeze (girls 60%, boys 48%) and previous wheeze (girls 33%, boys 28%) at baseline. In girls, development of current wheeze was significantly associated with current smoking (OR=2.8) and stable current wheeze was significantly associated with overweight (OR=2.4). Similar associations were not significant in boys. More girls than boys developed wheeze, had stable wheeze or had relapse of previous symptoms during the four year follow-up. The impact of smoking and overweight may put girls at a higher risk of respiratory symptoms than boys. Awareness of the gender difference in respiratory symptoms is important for diagnosis and preventive strategies during adolescence. PMID- 17084608 TI - Waste catalysts for waste polymer. AB - Catalytic cracking of high-density polyethylene (HDPE) over fluid catalytic cracking (FCC) catalysts (1:6 ratio) was carried out using a laboratory fluidized bed reactor operating at 450 degrees C. Two fresh and two steam deactivated commercial FCC catalysts with different levels of rare earth oxide (REO) were compared as well as two used FCC catalysts (E-Cats) with different levels of metal poisoning. Also, inert microspheres (MS3) were used as a fluidizing agent to compare with thermal cracking process at BP pilot plant at Grangemouth, Scotland, which used sand as its fluidizing agent. The results of HDPE degradation in terms of yield of volatile hydrocarbon product are fresh FCC catalysts>>steamed FCC catalysts approximately used FCC catalysts. The thermal cracking process using MS3 showed that at 450 degrees C, the product distribution gave 46 wt% wax, 14% hydrocarbon gases, 8% gasoline, 0.1% coke and 32% nonvolatile product. In general, the product yields from HDPE cracking showed that the level of metal contamination (nickel and vanadium) did not affect the product stream generated from polymer cracking. This study gives promising results as an alternative technique for the cracking and recycling of polymer waste. PMID- 17084609 TI - Microscale bioprocess optimisation. AB - Microscale processing techniques offer the potential to speed up the delivery of new drugs to the market, reducing development costs and increasing patient benefit. These techniques have application across both the chemical and biopharmaceutical sectors. The approach involves the study of individual bioprocess operations at the microlitre scale using either microwell or microfluidic formats. In both cases the aim is to generate quantitative bioprocess information early on, so as to inform bioprocess design and speed translation to the manufacturing scale. Automation can enhance experimental throughput and will facilitate the parallel evaluation of competing biocatalyst and process options. PMID- 17084610 TI - Electrical contacting of redox proteins by nanotechnological means. AB - Redox enzymes in bioelectronic devices usually lack direct electrical contact with electrodes, owing to the spatial separation of their redox centers from the conductive surfaces by the protein shells. The reconstitution of apo-enzymes on cofactor-functionalized nanostructures associated with electrodes provides a means to align the biocatalysts on the conductive surface and to electrically contact redox enzymes with electrodes. The reconstitution of apo-enzymes on cofactor-functionalized gold nanoparticles or carbon nanotubes has led to effective electrical communication between the redox proteins and the electrodes. Alternatively, the reconstitution of redox enzymes on molecular wires that enable electron tunneling or dynamic charge shuttling represent supramolecular biocatalytic nanostructures exhibiting electrical contact. The bioelectrocatalytic activities of the electrically wired reconstituted enzymes on electrodes have allowed the development of amperometric biosensors and biofuel cell elements. PMID- 17084611 TI - Nanoscale biocatalyst systems. AB - Since the large-scale application of immobilized enzymes in the 1960s, substantial research efforts have aimed to optimize the structure of carrier materials for better catalytic efficiency. In this regard, nanoscale materials provide the upper limits in balancing the key factors that determine the efficiency of biocatalysts, including surface area, mass transfer resistance, and effective enzyme loading. Various nanomaterials, such as nanoparticles, nanofibers, nanotubes and nanoporous matrices, have shown potential for revolutionizing the preparation and use of biocatalysts. Beyond their high surface area:volume ratios, nanoscale biocatalyst systems exhibit unique behaviors that distinguish them from traditional immobilized systems. The Brownian motion of nanoparticles, confining effect of nanopores and self assembling behaviors of discrete nanostructures are providing exciting opportunities in this field. The development of catalyst systems that are highly stable and efficient, capable of self-targeting or that function as molecular machines to catalyze multiple reactions is rapidly reshaping our vision of biocatalysts. PMID- 17084612 TI - Fragment-based lead discovery: a chemical update. AB - Fragment-based lead discovery constructs drug leads from small molecular fragments. In theory, this is a highly efficient method for drug discovery, and the technique has become enormously popular in the past few years. In this review, I describe how a variety of approaches in fragment-based lead discovery- including NMR, X-ray crystallography, mass spectrometry, functional screening, and in silico screening--have produced drug leads. Although the examples show that the technique can reliably generate potent molecules, there is still much work to be done to maintain the efficiency of molecules' binding affinities as fragments are linked, expanded, and otherwise improved. PMID- 17084613 TI - Insect olfactory memory in time and space. AB - Recent studies using functional optical imaging have revealed that cellular memory traces form in different areas of the insect brain after olfactory classical conditioning. These traces are revealed as increased calcium signals or synaptic release from defined neurons, and include a short-lived trace that forms immediately after conditioning in antennal lobe projection neurons, an early trace in dopaminergic neurons, and a medium-term trace in dorsal paired medial neurons. New molecular genetic tools have revealed that for normal behavioral memory performance, synaptic transmission from the mushroom body neurons is required only during retrieval, whereas synaptic transmission from dopaminergic neurons is required at the time of acquisition and synaptic transmission from dorsal paired medial neurons is required during the consolidation period. Such experimental results are helping to identify the types of neurons that participate in olfactory learning and when their participation is required. Olfactory learning often occurs alongside crossmodal interactions of sensory information from other modalities. Recent studies have revealed complex interactions between the olfactory and the visual senses that can occur during olfactory learning, including the facilitation of learning about subthreshold olfactory stimuli due to training with concurrent visual stimuli. PMID- 17084614 TI - Mechanisms for recovery of motor function following cortical damage. AB - Recent studies of focal injury to the cerebral cortex have demonstrated that the remaining, intact tissue undergoes structural and functional changes that could play a substantial role in neurological recovery. New information regarding the molecular and cellular environment in the adjacent, intact tissue has suggested that waves of growth promotion and inhibition modulate the self-repair processes of the brain. Furthermore, recent studies have documented widespread neurophysiological and neuroanatomical changes in regions remote from a focal cortical injury, suggesting that entire cortical networks participate in the recovery process. PMID- 17084615 TI - Basal ganglia oscillations and pathophysiology of movement disorders. AB - Low frequency rest tremor is one of the cardinal signs of Parkinson's disease and some of its animal models. Current physiological studies and models of the basal ganglia differ as to which aspects of neuronal activity are crucial to the pathophysiology of Parkinson's disease. There is evidence that neural oscillations and synchronization play a central role in the generation of the disease. However, parkinsonian tremor is not strictly correlated with the synchronous oscillations in the basal ganglia networks. Rather, abnormal basal ganglia output enforces abnormal thalamo-cortical processing leading to akinesia, the main negative symptom of Parkinson's disease. Parkinsonian tremor has probably evolved as a downstream compensatory mechanism. PMID- 17084616 TI - Representing episodes in the mammalian brain. AB - Memory lets the past inform the present so that we can attain future goals. In many species, these abilities require the hippocampus. Recent experiments, in which memory demand was varied while overt behavior and the environment were kept constant, have revealed firing patterns of hippocampal neurons that corresponded with memory demands and predicted performance. Although the active population appeared to be 'place cells' that signalled location, it actually included cells the activity patterns of which distinguished the recent or pending history of behavior during identical actions that occurred in the same place. Different populations of hippocampal cells fired as a rat walked along the same spatial path on the way to different goals, and coded past, present and pending events. Other experiments provide converging data that neuronal activity is modulated by goal-directed behavioral episodes. Together, these firing patterns suggest a testable mechanism of episodic memory coding: that hippocampal dynamics encode a temporally extended, hierarchically organized representation of goal-directed behavior. PMID- 17084617 TI - Prefrontal involvement in the regulation of emotion: convergence of rat and human studies. AB - Emotion regulation is a process by which we control when and where emotions are expressed. Paradigms used to study the regulation of emotion in humans examine controlled responses to emotional stimuli and/or the inhibition of emotional influences on subsequent behavior. These processes of regulation of emotion trigger activation of the ventromedial prefrontal cortex and inhibition of the amygdala. A similar pattern of activation is seen in rodents during recall of fear extinction, an example of emotional regulation. The overlap in circuitry is consistent with a common mechanism, and points toward future experiments designed to bridge human and rodent models of emotion regulation. PMID- 17084618 TI - Cellular principles underlying normal and pathological activity in the subthalamic nucleus. AB - The motor symptoms of Parkinson's disease are associated with abnormal, correlated, low frequency, rhythmic burst activity in the subthalamic nucleus and connected nuclei. Research into the mechanisms controlling the pattern of subthalamic activity has intensified because therapies that manipulate the pattern of subthalamic activity, such as deep brain stimulation and levodopa administration, improve motor function in Parkinson's disease. Recent findings suggest that dopamine denervation of the striatum and extrastriatal basal ganglia profoundly alters the transmission and integration of glutamatergic cortical and GABAergic pallidal inputs to subthalamic neurons, leading to pathological activity that resonates throughout the basal ganglia and wider motor system. PMID- 17084619 TI - Control strategies in object manipulation tasks. AB - The remarkable manipulative skill of the human hand is not the result of rapid sensorimotor processes, nor of fast or powerful effector mechanisms. Rather, the secret lies in the way manual tasks are organized and controlled by the nervous system. At the heart of this organization is prediction. Successful manipulation requires the ability both to predict the motor commands required to grasp, lift, and move objects and to predict the sensory events that arise as a consequence of these commands. PMID- 17084620 TI - Phase I study of continuous weekly dosing of dimethylamino benzoylphenylurea (BPU) in patients with solid tumours. AB - A phase I study of dimethylamino benzoylphenylurea (BPU), a tubulin inhibitor, was performed using a weekly continuous schedule. Patients with refractory solid tumours received oral BPU once weekly without interruption at doses ranging from 5 to 320mg using an accelerated titration design. Nineteen subjects received 54 cycles of BPU. Early pharmacokinetic findings of decreased clearance with increasing dose and plasma accumulation led to the expansion of the 320mg dose level. Two subjects then developed late haematologic dose-limiting toxicities (DLTs) that were associated with the highest plasma exposure to BPU and metabolites. Study enrollment resumed at dose 150mg with real-time pharmacokinetic monitoring. Seven additional subjects (6 evaluable) were treated for a median of 2 cycles (range 1.5-4) without further myelotoxicity. A long half life and accumulation of BPU and active metabolites were observed, recommending against a continuous administration. Weekly oral BPU therapy should be further tested using an interrupted schedule. PMID- 17084621 TI - Randomised trial of sequential versus concurrent chemo-radiotherapy in patients with inoperable non-small cell lung cancer (EORTC 08972-22973). AB - A randomised phase III study was performed comparing sequential (S) and concurrent (C) chemo-radiotherapy (CRT) in non-small cell lung cancer (NSCLC) patients. METHODS: One hundred and fifty-eight patients were randomised to receive two courses of Gemcitabine (1250mg/m(2) days 1, 8) and Cisplatin (75mg/m(2) day 2) prior to, or daily low-dose Cisplatin (6mg/m(2)) concurrent with radiotherapy, consisting of 24 fractions of 2.75Gy in 32 days, with a total dose of 66Gy. RESULTS: Acute haematological toxicity grade 3/4 was more pronounced in the sequential (S) (30% versus 6%), oesophagitis grade 3/4 more frequent in the concurrent (C) arm (5% versus 14%). Late oesophagitis grade 3 was 4% (S and C), pneumonitis grade 3/4 14% (S) and 18% (C). Because of the poor power of the study no significant differences in median survival (MS), overall survival (OS) and progression-free survival (PFS) could be detected. MS was 16.2 (S) and 16.5 (C) months, 2-year OS was 34% (S) and 39% (C), 3-year OS was 22% (S) and 34% (C). CONCLUSION: Radiotherapy 66Gy given concurrently with daily low-dose Cisplatin or after two courses of Gemcitabine/Cisplatin was well tolerated. Due to early closure no conclusions can be reached on the relative merits; both arms showed good OS. PMID- 17084622 TI - Survival of cancer patients in France: a population-based study from The Association of the French Cancer Registries (FRANCIM). AB - We present the main results of the first population-based cancers survival study gathering all French registry data. Survival data on 205,562 cancer cases diagnosed between 01/01/1989 and 31/12/1997 were analysed. Relative survival was estimated using an excess rate model. The evolution of the excess mortality rate over the follow-up period was graphed. The analysis emphasised the effect of age at diagnosis and its variation with time after diagnosis. For breast and prostate cancers, the age-standardised five-year relative survivals were 84% and 77%, respectively. The corresponding results in men and women were 56% versus 58% for colorectal cancer and 12% versus 16% for lung cancer. For some cancer sites, the excess mortality rate decreased to low values by five years after diagnosis. For most cancer sites, age at diagnosis was a negative prognostic factor but this effect was often limited to the first year after diagnosis. PMID- 17084623 TI - The role of glutathione-S-transferase polymorphisms in ovarian cancer survival. AB - Resistance to chemotherapy represents one of the most important causes of treatment failure in patients with ovarian cancer. Common polymorphisms in the glutathione-S-transferase (GSTM1, GSTP1 and GSTT1) family have been implicated in chemoresistence and ovarian cancer survival. In this study, we have analysed Australian women diagnosed with primary invasive epithelial ovarian cancer between 1985 and 1997, using DNA extracted from peripheral blood and archival uninvolved (normal) tissues. GSTP1 genotypes were determined using ABI Prism 7700 Sequence Detection System methodology (n=448) and GSTT1 and GSTM1 genotypes using PCR-agarose methodology (n=239). We observed a significant survival advantage among carriers of GSTP1 Ile105Val GG/GA genotype (HR 0.77, 95% confidence interval (CI) 0.61-0.99,p=0.04) and a non-significant survival advantage among women who were homozygous for the GSTM1 and GSTT1 deletion variants. There was also evidence of an additive effect, with a stronger survival benefit in women carrying three low function GST genotypes (GSTM1 null, GSTT1 null and GSTP1 GA/GG) (HR 0.47, 95% CI 0.22-1.02). The results of this study, the largest to date, are consistent with a number of previous smaller studies which have also observed that reduced GST function was associated with better survival outcomes in patients with ovarian cancer. PMID- 17084624 TI - Rapid endocrine disruption: environmental estrogen actions triggered outside the nucleus. AB - An exogenous substance is defined as an endocrine disrupter chemical (EDC) if it alters the function of the endocrine system provoking adverse health effects. Environmental estrogens are the most studied EDCs. They follow the same mechanisms of action as the gonadal hormone 17beta-estradiol. Up to now, the estrogenicity of environmental estrogenic pollutants has been based on the property of these compounds to bind to estrogen receptors (ERs), either ERalpha or ERbeta, and to act subsequently as transcription factors when binding to the estrogen response element (ERE) in the DNA. All the estrogenic bioassays currently used are based on this mechanism of action. New evidence indicates that the definition of estrogenicity for a chemical should take into account other estrogen receptors as well as new signaling pathways. These include the activation of additional transcription factors as well as the action of xenoestrogens through estrogen receptors located outside the nucleus: in the plasma membrane, mitochondria and probably the cytosol. Therefore, new estrogenic bioassays should be developed to include the novel concept of rapid endocrine disruption. PMID- 17084625 TI - Metabolism of DHEA in postmenopausal women following percutaneous administration. AB - The marked decline in serum dehydroepiandrosterone (DHEA) with age is believed to play a role in health problems associated with aging, these health issues being potentially preventable or reversible by the exogenous administration of DHEA. In the present study, liquid chromatography/mass spectrometry/mass spectrometry (LC/MS/MS) and gas chromatrography/mass spectrometry (GC/MS) were used to measure the serum levels of DHEA and 11 of its metabolites in seventy-five 60-65-year-old Caucasian women who received 3g of 0.1%, 0.3%, 1.0% or 2.0% DHEA cream or placebo applied twice daily on the face, upper chest, arms and legs. The serum levels of DHEA increased 574% over control at the 2.0% DHEA dose while the sum of the androgen metabolites androsterone glucuronide (ADT-G), 3alpha-androstenediol-3G (3alpha-diol-3G) and 3alpha-diol-17G increased by only 231%. On the other hand, serum testosterone and dihydrosterone were increased by 192% and 275%, respectively, above basal levels compared to 139% and 158% for estrone and estradiol. Such data show that the transformation of exogenous DHEA in postmenopausal women is preferentially into androgens rather than into estrogens. On the other hand, the present data indicate that serum DHEA measurements following DHEA supplementation in postmenopausal women are an overestimate of the formation of active androgens and estrogens and suggest a decreased efficiency of transformation of DHEA into androgens and estrogens with aging. PMID- 17084626 TI - A full conformational characterization of 13-ethylprogestogens through theoretical calculations and nuclear magnetic resonance spectroscopy. AB - The conformational preferences of a group of 13-ethylsteroids and related estranes have been determined through theoretical calculations at the B3LYP/6 31G(*) level in order to ascertain differences and similarities in the conformational behavior which might, in principle, influence the activity. Attention was focussed on two geometrical features usually related to the progestational activity of this class of compounds, namely, the inversion of the A ring and the orientation of the 13-ethyl group. The present calculations show that the absence of the C10 methyl group, like in levonorgestrel, 3 ketodesogesterel, and gestodene, makes the inversion of the A ring easier than in norethisterone and ethisterone even if in any case the 1alpha,2beta-half-chair conformation remains preferred. The preference of the ethyl group for the trans orientation is maintained by all the 13-ethylprogestogens though in gestodene also the g(+)gauche conformation is significantly populated. This preference was experimentally supported through a high field NMR spectroscopy study of the ethylsteroids. PMID- 17084627 TI - Design and synthesis of substrate-mimic inhibitors of mycothiol-S-conjugate amidase from Mycobacterium tuberculosis. AB - The Staudinger reaction between a polymer-supported triphenylphosphine reagent and pseudo-disaccharide azides is successfully applied to synthesize a variety of substrate-mimic mycothiol analogs. Screening of this new group of analogs against the mycobacterial detoxification enzyme mycothiol-S-conjugate amidase (MCA) yielded several modest inhibitors (IC50 values around 50 microM) and provided additional structure-activity relationships for future optimization of inhibitors of MCA and its homologs. PMID- 17084628 TI - Coping with stress: cellular relaxation techniques. AB - Proteins damaged by stressors such as heat, oxidizing conditions or toxic agents are deleterious to cells and must be properly taken care of. Accordingly, misfolded proteins trigger a cellular stress response that aims to either repair defective polypeptides or eliminate faulty elements when salvage is not possible. This stress response provides time for additional stressor-specific pathways that adapt the cell to the changed environment if necessary. Recent studies have investigated how proteins that frustrate the folding machinery are recognized and cleared from the cell. Surprisingly, these clearance mechanisms are not restricted to the protein level. The stress response can also eliminate the mRNA of polypeptides that are refractory to folding. PMID- 17084630 TI - The influence of artificially increased trunk stiffness on the balance recovery after a trip. AB - Falls occur frequently in the growing population of elderly. Since trunk control is critical for maintaining balance, the higher trunk stiffness in elderly people compared to the general population has been associated with their increased fall risk. Theoretically, trunk stiffness may be beneficial for balance recovery in walking, i.e. after a trip. A stiff joint may provide a torque that restricts the perturbation effects and thereby reduces the probability of a fall. The aim of this study was to test whether trunk stiffness impaired or assisted balance recovery after a trip during walking. An orthopedic corset was used to simulate trunk stiffness in 11 young male adults. Subjects walked over a platform, with or without the corset on, and were occasionally tripped over a hidden obstacle. Kinematics of the tripping reaction were measured. Initial trunk accelerations were significantly attenuated by the corset, which indicates a positive effect of the stiffening corset. However, no subsequent effects on peak trunk inclination and on the peak moment arm of gravity on the trunk were found. The pattern of trunk motion allowed ample time for triggered or voluntary muscle responses to be generated, before a substantial inclination occurred. It appears that such active responses were sufficient in the young subjects tested to obtain a similar net effect with or without the increased trunk stiffness induced by the corset. PMID- 17084631 TI - Identifying interaction motifs in CK2beta--a ubiquitous kinase regulatory subunit. AB - Casein kinase 2 (CK2) is probably the most ubiquitous serine/threonine kinase found in eukaryotes: it phosphorylates >300 cellular proteins, ranging from transcription factors to proteins involved in chromatin structure and cell division. CK2 is a heterotetrameric enzyme that induces neoplastic growth when overexpressed. The beta subunit of CK2 (CK2beta) functions as the regulator of the catalytic CK2alpha and CK2alpha' subunits, enhancing their stability, activity and specificity. However, CK2beta also functions as a multisubstrate docking platform for several other binding partners. Here, we discuss the organization and roles of interaction motifs of CK2beta, postulate new protein interaction sites and map these to the known interaction motifs, and show how the resulting complexity of interactions mediated by CK2 gives rise to the versatile functions of this pleiotropic protein kinase. PMID- 17084632 TI - Pathways for mRNA localization in the cytoplasm. AB - Studies of the intracellular localization of mRNA have clearly demonstrated that certain subsets of mRNA are concentrated in discrete locations within the cytoplasm. Localization is one aspect of the post-transcriptional control of gene expression, and is intertwined with the translation and turnover of mRNA to achieve the goal of local protein production. Different mechanisms have been identified that enable localized mRNAs to target different subcellular compartments, and recent advances in understanding these pathways is reviewed here. PMID- 17084633 TI - Synthesis, antimalarial, antileishmanial, antimicrobial, cytotoxicity, and methemoglobin (MetHB) formation activities of new 8-quinolinamines. AB - We report the synthesis, in vitro antiprotozoal (against Plasmodium and Leishmania), antimicrobial, cytotoxicity (Vero and MetHb-producing properties), and in vivo antimalarial activities of two series of 8-quinolinamines. N1-{4-[2 (tert-Butyl)-6-methoxy-8-quinolylamino]pentyl}-(2S/2R)-2-aminosubstitutedamides (21-33) and N1-[4-(4-ethyl-6-methoxy-5-pentyloxy-8-quinolylamino)pentyl]-(2S/2R) 2-aminosubstitutedamides (51-63) were synthesized in six steps from 6-methoxy-8 nitroquinoline and 4-methoxy-2-nitro-5-pentyloxyaniline, respectively. Several analogs displayed promising antimalarial activity in vitro against Plasmodium falciparum D6 (chloroquine-sensitive) and W2 (chloroquine-resistant) clones with high selectivity indices versus mammalian cells. The most promising analogs (21 24) also displayed potent antimalarial activity in vivo in a Plasmodium berghei infected mouse model. Most interestingly, many analogs exhibited promising in vitro antileishmanial activity against Leishmania donovani promastigotes, and antimicrobial activities against a panel of pathogenic bacteria and fungi. Several analogs, notably 21-24, 26-32, and 60, showed less MetHb formation compared to primaquine indicating the potential of these compounds in 8 quinolinamine-based antimalarial drug development. PMID- 17084634 TI - Dioxane and oxathiane nuclei: suitable substructures for muscarinic agonists. AB - Muscarinic agonists, bearing 1,4-dioxane and 1,4-oxathiane nuclei, were synthesized and tested to evaluate their potency at M(1)-M(4) muscarinic receptor subtypes. The stereochemical relationship between the 2-side chain and the 6 methyl group plays an important role in drug-receptor interaction, since the cis isomers are more potent than the corresponding trans isomers. However, the latter are able to discriminate between the muscarinic receptor subtypes. Among them compound 5b proves particularly interesting, since it selectively activates the ileal M(3) receptor subtype and is devoid of agonist activity at the others. PMID- 17084635 TI - Inhibition of free radical-mediated oxidation of cellular biomolecules by carboxylated chitooligosaccharides. AB - The objective of this study was to identify the cellular antioxidant effects of carboxylated chitooligosaccharides (CCOS), a chemically modified derivative of chitooligosaccharides (COS), by assessing oxidation inhibition potential on cellular biomolecules such as lipids, proteins, and direct scavenging of reactive oxygen species (ROS). Radical-mediated oxidation of cell membrane lipids and proteins was dose-dependently inhibited by CCOS, assessed by amount of lipid hydroperoxides and carbonyl carbon content in mouse macrophages, RAW264.7 cells. Further, CCOS inhibited myeloperoxidase (MPO) activity in human myeloid cells (HL60) suggesting indirect possibility of inhibiting generation of reactive oxygen species (ROS) such as superoxide radicals, H(2)O(2) and HOCl. Direct radical scavenging studies carried out with DCFH-DA fluorescence probe concluded that CCOS can act as a potent radical scavenger in cells. PMID- 17084636 TI - Measurement of the spatial response of a detector pixel. AB - In order to measure the internal spatial response of a pixel in a detector, it is scanned by a beam smaller than its size. This becomes difficult as the wave length grows and becomes comparable to the pixel size, such as in the infra red. To overcome this difficulty, a special phase mask which makes the beam narrower was designed, constructed, and tested successfully. The mask was made from five alternating transparent rings, where the rings had half a wave phase difference between them. The beam was scanned with and without the mask in two dimensions in fine steps by a much smaller detector and its response was taken. The spot width dropped by 19% at half its height and by 42% at tenth its height, a significant narrowing. The scan was repeated with the full detector pixel. That beam scan served as a deconvolution kernel and allowed us to find the pixel point spread function (spatial response), the pixel modulation transfer function and the optical cross talk between the pixels. PMID- 17084637 TI - Cloning and characterization of a cowpea seed lipid transfer protein cDNA: expression analysis during seed development and under fungal and cold stresses in seedlings' tissues. AB - Lipid transfer proteins (LTPs) are antimicrobial peptides (AMPs) involved in the defense of plants against pathogens. Our group has previously characterized and purified a LTP from cowpea (Vigna unguiculata (L.) Walp.) seeds which caused the inhibition of growth of fungal pathogens in vitro. The aim of this work was to obtain the cDNA encoding the cowpea LTP and after cloning, to use the cDNA as a probe for studying its expression profile during the development of cowpea seeds. In this work, the N-terminal sequence of the mature LTP peptide from cowpea was used to produce a degenerated oligonucleotide. This primer allowed the amplification of the LTP cDNA by RT-PCR from mRNA of cowpea seeds. The sequence analysis of the cloned cDNA, named VULTP, showed 494 bp which encoded a polypeptide of 91 amino acids. The deduced peptide presented high homology of similarity to plant LTPs of Vigna radiata var. radiate (94%), Prunus domestica (82%) and Zea mays (72%). The expression profile of the VULTP gene in cowpea was analyzed by Northern blot and revealed that the transcript is not accumulated in adult tissues. Conversely, VULTP mRNA is early and strongly accumulated during seed development. The results obtained to seedling of cowpea demonstrate that the VULTP gene presents differential expression in response to different stress. Further studies will be conducted to try to gain better understanding about the physiological role of this gene in cowpea. PMID- 17084638 TI - Resolution of the excitation-emission spectra of FMN in rigid poly(vinyl alcohol) matrices. AB - The excitation-emission spectra of flavin mononucleotide (FMN) were measured in rigid PVA films for concentrations ranging from 6.92 x 10(-4)M to 1.03 M. The theoretical three-linear decomposition of the excitation-emission spectra indicated the presence of two absorption and emission centers corresponding to FMN monomer and dimer, respectively. The component of the fluorescence profile corresponding to the FMN monomer has a large negative part which is the mirror image of the emission band profile of the dimer. The elimination of this part by taking a linear combination of the emission components of the monomer and of the dimer resulted in emission spectrum, which is in a very good agreement with the monomer spectrum measured directly. The appearance of a negative part of the monomer emission profile obtained by trilinear decomposition of the emission absorption spectra of FMN can be explained in terms of the non-radiative reverse energy transfer from the FMN dimers to the FMN monomers. The presented results confirm that the FMN molecules in rigid PVA form dimers but not higher order aggregates. Moreover, they enable to obtain fluorescence spectra of dimers and suggest that FMN dimers may take part in the process of non-radiative energy transfer occurring in photoreception phenomena. PMID- 17084639 TI - The adipocyte-myocyte axis in insulin resistance. AB - Insulin resistance in skeletal muscle is linked to an elevated adipose tissue mass, as is found in obesity, but can also be observed in lipodystrophy, in which adipose tissue is greatly reduced. Adipose tissue releases endocrine and metabolic mediators and is actively involved in crosstalk with skeletal muscle, a process that precedes and underlies the development of insulin resistance in muscles. Adipokines including tumor necrosis factor alpha, interleukin-6, leptin and adiponectin influence insulin signaling in skeletal muscle. Free fatty acids, their metabolites and ectopic fat in muscle also contribute to insulin resistance. Recent research indicates inflammation, endoplasmic reticulum stress and oxidative stress could be underlying mechanisms at the center of the development of insulin resistance. Insights into the role of macrophages in adipose tissue add to the complicated interplay between adipose tissue and skeletal muscle. PMID- 17084640 TI - Mechanisms of peptide hormone secretion. AB - According to the classical view, peptide hormones are stored in large dense-core vesicles that release all of their cargo rapidly and completely when they fuse with and flatten into the plasma membrane. However, recent imaging studies suggest that this view is too simple. Even after vesicles fuse with the plasma membrane, cells might control the rate of dispersal of vesicle cargo - either by modulating the properties of the fusion pore that connects the vesicle lumen to the extracellular solution or by storing cargo in states that disperse slowly in the extracellular space. Understanding these mechanisms is important, owing to the increasing prevalence of diseases, such as type 2 diabetes mellitus, which arise from insufficient secretion of peptide hormones. PMID- 17084641 TI - Protein phosphatase 5 as a negative key regulator of Raf-1 activation. AB - Growth factors such as the epidermal growth factor cause sequential activation of receptor tyrosine kinases, adaptor molecules and the Raf-MEK-ERK pathway. The kinetics and intensity of these signals are dependent on the balance between phosphorylation and dephosphorylation of these molecules by numerous kinases and phosphatases, respectively. Recently, protein phosphatase 5 has been characterized as a key dephosphorylation regulator of Raf-1 activation in growth factor-mediated signaling, leading to attenuation of the MEK-ERK cascade. PMID- 17084642 TI - Isolation, purification and characterization of a novel glucose oxidase from Penicillium sp. CBS 120262 optimally active at neutral pH. AB - A novel glucose oxidase (GOX), a flavoenzyme, from Penicillium sp. was isolated, purified and partially characterised. Maximum activities of 1.08U mg(-1)dry weight intracellular and 6.9U ml(-1) extracellular GOX were obtained. Isoelectric focussing revealed two isoenzymes present in both intra- and extracellular fractions, having pI's of 4.30 and 4.67. GOX from Penicillium sp. was shown to be dimeric with a molecular weight of 148kDa, consisting of two equal subunits with molecular weight of 70k Da. The enzyme displayed a temperature optimum between 25 and 30 degrees C, and an optimum pH range of 6-8 for the oxidation of beta-d glucose. The enzyme was stable at 25 degrees C for a minimum of 10h, with a half life of approximately 30 min at 37 degrees C without any prior stabilisation. The lyophilized enzyme was stable at -20 degrees C for a minimum of 6 months. GOX from Penicillium sp. Tt42 displayed the following kinetic characteristics: Vmax, 240.5U mg(-1); Km, 18.4mM; kcat, 741 s(-1) and kcat/Km, 40 s(-1)mM(-1). Stability at room temperature, good shelf-life without stabilisation and the neutral range for the pH optimum of this GOX contribute to its usefulness in current GOX-based biosensor applications. PMID- 17084644 TI - An improved molecular phylogeny of the Nematoda with special emphasis on marine taxa. AB - Phylogenetic reconstructions of relations within the phylum Nematoda are inherently difficult but have been advanced with the introduction of large-scale molecular-based techniques. However, the most recent revisions were heavily biased towards terrestrial and parasitic species and greater representation of clades containing marine species (e.g. Araeolaimida, Chromadorida, Desmodorida, Desmoscolecida, Enoplida, and Monhysterida) is needed for accurate coverage of known taxonomic diversity. We now add small subunit ribosomal DNA (SSU rDNA) sequences for 100 previously un-sequenced species of nematodes, including 46 marine taxa. SSU rDNA sequences for >200 taxa have been analysed based on Bayesian inference and LogDet-transformed distances. The resulting phylogenies provide support for (i) the re-classification of the Secernentea as the order Rhabditida that derived from a common ancestor of chromadorean orders Araeolaimida, Chromadorida, Desmodorida, Desmoscolecida, and Monhysterida and (ii) the position of Bunonema close to the Diplogasteroidea in the Rhabditina. Other, previously controversial relationships can now be resolved more clearly: (a) Alaimus, Campydora, and Trischistoma belong in the Enoplida, (b) Isolaimium is placed basally to a big clade containing the Axonolaimidae, Plectidae, and Rhabditida, (c) Xyzzors belongs in the Desmodoridae, (d) Comesomatidae and Cyartonema belongs in the Monhysterida, (e) Globodera belongs in the Hoplolaimidae and (f) Paratylenchus dianeae belongs in the Criconematoidea. However, the SSU gene did not provide significant support for the class Chromadoria or clear evidence for the relationship between the three classes, Enoplia, Dorylaimia, and Chromadoria. Furthermore, across the whole phylum, the phylogenetically informative characters of the SSU gene are not informative in a parsimony analysis, highlighting the short-comings of the parsimony method for large-scale phylogenetic modelling. PMID- 17084643 TI - Phosphorylation and activation of a transducible recombinant form of human HSP20 in Escherichia coli. AB - Protein-based cellular therapeutics have been limited by getting molecules into cells and the fact that many proteins require post-translational modifications for activation. Protein transduction domains (PTDs), including that from the HIV TAT protein (TAT), are small arginine rich peptides that carry molecules across the cell membrane. We have shown that the heat shock-related protein, HSP20 is a downstream-mediator of cyclic nucleotide-dependent relaxation of vascular smooth muscle and is activated by phosphorylation. In this study, we co-expressed in Escherichia coli the cDNAs encoding the catalytic subunit of protein kinase G and a TAT-HSP20 fusion protein composed of the TAT PTD (-YGRKKRRQRRR-) fused to the N terminus of human HSP20. Immunoblot and HPLC-ESI-MS/MS analysis of the purified TAT-HSP20 demonstrated that it was phosphorylated at serine 40 (equivalent to serine 16 in wild-type human HSP20). This phosphorylated TAT-HSP20 was physiologically active in intact smooth muscles in that it inhibited 5 hydroxytryptamine-induced contractions by 57%+/-4.5. The recombinant phosphorylated protein also led to changes in actin cytoskeletal morphology in 3T3 cells. These results delineate strategies for the expression and activation of therapeutic molecules for intracellular protein based therapeutics. PMID- 17084645 TI - Distinguishing gorilla mitochondrial sequences from nuclear integrations and PCR recombinants: guidelines for their diagnosis in complex sequence databases. AB - Nuclear integrations of mitochondrial DNA (Numts) are widespread in many taxa and if left undetected can confound phylogeny interpretation and bias estimates of mitochondrial DNA (mtDNA) diversity. This is particularly true in gorillas, where recent studies suggest multiple integrations of the first hypervariable (HV1) domain of the mitochondrial control region. Problems can also arise through the inadvertent incorporation of artifacts produced by in vitro recombination between sequence types during polymerase chain reaction amplification. This issue has attracted little attention yet could potentially exacerbate errors in databases already contaminated by Numts. Using a set of existing diagnostic tools, this study set out to systematically inventory Numts and PCR recombinants in a gorilla HV1 sequence database and address the degree to which existing public databases are contaminated. Phylogenetic analysis revealed three distinct gorilla HV1 Numt groups (I, II, and III) that could be readily differentiated from mtDNA sequences by Numt-specific diagnostic sites and sequence-based motifs. Several instances of genuine recombination were also identified by a suite of detection methods. The location of putative breakpoints was identified by eye and by likelihood analysis. Findings from this study reveal widespread nuclear contamination of gorilla HV1 GenBank databases and underline the importance of recognizing not only Numts but also PCR recombinant artifacts as potential sources of data contamination. Guidelines for the routine identification of Numts and in vitro recombinants are presented and should prove useful in the detection of similar artifacts in other species mtDNA databases. PMID- 17084646 TI - Immune responses in mice to arecoline mediated by lymphocyte muscarinic acetylcholine receptor. AB - There is evidence that lymphocytes possess all the components of the cholinergic system independent of neuronal innervations. Thus, potential therapeutic applications of drugs targeting the neuronal cholinergic system might have side effects on the immune system. This study investigated whether arecoline could affect immunological functions in mice and explored the mechanism of the effect of arecoline on the immune system. To investigate this, arecoline at the dose of 2mg/kg was administered subcutaneously in BALB/c mice for 4 weeks to evaluate changes in immunological function both in vivo and in vitro. Several indices were used to assess immunological activation, including the spleen index, serum hemolysin levels, interleukin (IL)-2 and splenocyte proliferation. Our results showed a significant reduction in treated animals with respect to the control group in the following tests: the spleen index (86%), hemolysin against sheep red blood cells (68%), IL-2 production (73%), and splenocyte proliferation induced by concanavalin A or lipopolysaccharide (76% and 74%, respectively). The muscarinic receptor antagonist atropine (1mg/kg) reversed the inhibition of the four immune related parameters mentioned above. Chronic atropine alone did not significantly affect the immune response. To our knowledge, this is the first study to demonstrate that arecoline interferes with the immune system by targeting the muscarinic acetylcholine receptors of the non-neuronal cholinergic system. PMID- 17084647 TI - Learned preferences induced by electrical stimulation of a food-related area of the parabrachial complex: effects of naloxone. AB - Electrical stimulation of the External Lateral Parabrachial Subnucleus (LPBe), a food-related area, induced behavioral preferences for associated stimuli in a taste discrimination learning task. Although this stimulation appeared to be ineffective to elicit standard lever press self-stimulation, it induced place preference for one of two training compartments of a rectangular maze in which animals (adult male Wistar rats) received concurrent electrical brain stimulation. In subjects that consistently showed a preference behavior in different trials, administration of the opioid antagonist naloxone (4 mg/ml/kg) blocked concurrent learning when the test was made in a new maze but not in the same maze in which animals had learned the task. These results are discussed in terms of the possible participation of the LPBe subnucleus in different natural and artificial brain reward systems. PMID- 17084648 TI - Sleep and sleep disorders in adults with attention deficit/hyperactivity disorder. AB - Attention deficit/hyperactivity disorder (ADHD) is a frequent disorder that often persists in adulthood. Persistent ADHD is known to be a serious risk factor for other disorders in adulthood, and adults with ADHD often report on sleep disorders. Despite this, only few studies have investigated the subjective and objective quality of sleep in adults suffering from ADHD. Previous studies have revealed seriously impaired subjective sleep quality and increased nocturnal motor activity in spite of essentially normal standard polysomnographic parameters in this patient group. However, primary sleep disorders such as sleep apnea syndrome or restless legs syndrome (RLS) may be misdiagnosed as ADHD. Moreover, ADHD and primary sleep disorders may occur as comorbidities. In particular, RLS was suggested to be highly associated with ADHD, indicating a probable common central nervous dopaminergic dysfunction. To date, larger studies with adequate sample sizes that compare sleep in adult patients with ADHD, healthy control groups and patients with other primary sleep disorders are still lacking. PMID- 17084649 TI - Computed tomography of the vertebral column and coelomic structures in the normal loggerhead sea turtle (Caretta caretta). AB - The aim of this study was to determine the normal computed tomography (CT) appearance of the vertebral column and coelomic structures of the loggerhead sea turtle (Caretta caretta) and to use three-dimensional (3D) and multiplanar reconstructions to indicate the position of each organ in relation to the vertebrae and carapace. Transverse sections of 1mm thickness were performed in seven clinically healthy and in five dead loggerhead sea turtles using multi detector CT equipment. A computer workstation was used for multiplanar and 3D reconstructions. Dead turtles were frozen and sectioned in the transverse, dorsal and sagittal planes to compare the anatomical structures' appearance with CT images. Clinically relevant organs including the oesophagus, stomach, trachea, bronchi, lungs, liver, gallbladder, heart, spleen, kidneys and vertebral canal were identified in CT images. Computed tomography provides detailed information on the respiratory system and skeleton; the location of the coelomic structures with respect to the carapace and the vertebrae that is provided in this work will facilitate the use of other ancillary diagnostic techniques such as ultrasound, radiography and biopsy, thereby improving safety of access in surgical procedures. PMID- 17084650 TI - Trehalose-protected lipid membranes for determining membrane protein structure and insertion. AB - Trehalose preserves lipid bilayers during dehydration and rehydration by replacing water to form hydrogen bonds between its own OH groups and lipid headgroups. We compare the lipid conformation and dynamics between trehalose protected lyophilized membranes and hydrated membranes, to assess the suitability of the trehalose-containing membrane as a matrix for membrane protein structure determination. (31)P spectra indicate that the lipid headgroup of trehalose protected dry POPC membrane (TRE-POPC) have an effective phase transition temperature that is approximately 50K higher than that of the hydrated POPC membrane. In contrast, the acyl chains have similar transition temperatures in the two membranes. Intramolecular lipid (13)C'-(31)P distances are the same in TRE-POPC and crystalline POPC, indicating that the lipid headgroup and glycerol backbone conformation is unaffected by trehalose incorporation. Intermolecular (13)C-(31)P distances between a membrane peptide and the lipid headgroups are 10% longer in the hydrated membrane at 226 K than in the trehalose-protected dry membrane at 253 K. This is attributed to residual motions in the hydrated membrane, manifested by the reduced (31)P chemical shift anisotropy, even at the low temperature of 226 K. Thus, trehalose lyoprotection facilitates the study of membrane protein structure by allowing experiments to be conducted at higher temperatures than possible with the hydrated membranes. PMID- 17084651 TI - 2D separated-local-field spectra from projections of 1D experiments. AB - A novel procedure for reconstruction of 2D separated-local-field (SLF) NMR spectra from projections of 1D NMR data is presented. The technique, dubbed SLF projection reconstruction from one-dimensional spectra (SLF-PRODI), is particularly useful for uniaxially oriented membrane protein samples and represents a fast and robust alternative to the popular PISEMA experiment which correlates (1)H-(15)N dipole-dipole couplings with (15)N chemical shifts. The different 1D projections in the SLF-PRODI experiment are obtained from 1D spectra recorded under influence of homonuclear decoupling sequences with different scaling factors for the heteronuclear dipolar couplings. We demonstrate experimentally and numerically that as few as 2-4 1D projections will normally be sufficient to reconstruct a 2D SLF-PRODI spectrum with a quality resembling typical PISEMA spectra, leading to significant reduction of the acquisition time. PMID- 17084652 TI - Bloch-Redfield-Wangsness theory engine implementation using symbolic processing software. AB - We describe a general method for the automated symbolic processing of Bloch Redfield-Wangsness relaxation theory equations for liquid-phase spin dynamics in the algebraically challenging case of rotationally modulated interactions. The processing typically takes no more than a few seconds (on a contemporary single processor workstation) and yields relaxation rate expressions that are completely general with respect to the spectral density functions, relative orientations, and magnitudes of the interaction tensors, with all cross-correlations accounted for. The algorithm easily deals with fully rhombic interaction tensors, and is able, with little if any modification, to treat a large variety of the relaxation mechanisms encountered in NMR, EPR, and spin dynamics in general. PMID- 17084653 TI - Therapy for Gaucher disease: don't stop thinking about tomorrow. AB - While enzyme replacement therapy for Gaucher disease has been widely used and appears to be an efficacious and safe treatment, this success should not be a reason for complacency. Other treatment strategies currently under consideration for patients with Gaucher disease include gene therapy, substrate reduction therapy and chaperone therapy. Furthermore, improvements in enzyme therapy could also have a significant clinical impact. Individuals with Gaucher disease and other lysosomal disorders will greatly benefit from continual refinement and optimization of the current therapy, as well as from the development of new treatment modalities that offer improvements in efficacy, cost, safety and availability. PMID- 17084654 TI - Absorption kinetics of ozone in water with ultrasonic radiation. AB - A mathematical model was proposed to depict classical unsteady state method that was used to determine volumetric mass transfer coefficient of ozone from gaseous phase to aqueous phase during sonolysis. The rate constant of ozone self decomposition with ultrasonic radiation, which was one of the parameters in the model, was determined with separate experiments. The results showed that self decomposition rate constants of ozone were enhanced by ultrasound. The self decomposition rate constant of ozone is linearly dependent on ultrasonic power, but the increase of the decomposition rate could not enhance ozone mass transfer coefficient. The volumetric mass transfer coefficients of ozone were also enhanced by ultrasonic radiation, while ultrasonic power had little effect on volumetric mass transfer coefficient of ozone. The degassing effect of ozone due to ultrasonic radiation was insignificant in the sparged system when ozone was bubbled during sonolysis. PMID- 17084655 TI - Neighborhood social capital and adult health: an empirical test of a Bourdieu based model. AB - Drawing upon Bourdieu's [1986. The forms of capital. In: Richardson, J.G. (Ed.), Handbook of Theory and Research for the Sociology of Education. Greenwood, New York, pp. 241-258.] social capital theory, I test a conceptual model of neighborhood conditions and social capital - considering relationships between neighborhood social capital forms (social support, social leverage, informal social control, and neighborhood organization participation) and adult health behaviors (smoking, binge drinking) and perceived health, as well as interactions between neighborhood social capital and individuals' access to that social capital. Analyzing Los Angeles Family and Neighborhood Survey data linked with tract level census data, results suggest that specific social capital forms were directly associated with both positive and negative health outcomes. Additionally, residents' neighborhood attachment moderated relationships between various social capital forms and health. Future studies should consider social capital resources and the role of differential access to such resources for promoting or compromising health. PMID- 17084656 TI - Identification of hydrophobic proteins as biomarker candidates for colorectal cancer. AB - Nowadays, colorectal cancer is one of the major causes of cancer death in Western countries. Due to the lack of biomarkers with clinical utility for this pathology, and considering that membrane and hydrophobic proteins have not been studied in depth, we performed a prefractionation of colorectal tissues prior to two-dimensional gel electrophoresis in order to identify hydrophobic proteins differentially expressed in colorectal cancer patients. Fractions enriched in hydrophobic proteins were obtained from healthy mucosa and tumor tissue by a specific extraction method based on temperature-dependent phase partitioning with Triton X-114. Proteins were separated by two-dimensional gel electrophoresis and gels were silver-stained, scanned and compared using the PDQuest software. Those spots presenting significantly different abundance were submitted to mass spectrometry for protein identification. Alterations in the expression of cytoskeletal proteins, including a decrease of vimentin and the absence of desmin, were found. We also detected alterations in antioxidant and transport proteins, chaperones, and in two isoforms of the calcium-binding protein S100A6. On the other hand, vimentin was chosen to corroborate the electrophoretic results by specific immunodetection. Most of the altered proteins have been related to cellular membranes, many of them to lipid rafts microdomains in the plasma membrane, and they have also been implicated in the control of cell proliferation, apoptosis, or metastasis. In conclusion, all the proteins found altered in colorectal tumor samples could be considered as candidates for future studies focused on their utility as markers for colorectal diagnosis and prognosis, or as targets for colorectal cancer therapy. PMID- 17084657 TI - Identification and expression of a novel class of glutathione-S-transferase from amphioxus Branchiostoma belcheri with implications to the origin of vertebrate liver. AB - Glutathione-S-transferases have been identified in all the living species examined so far, yet little is known to date about them in amphioxus, a model organism for insights into the origin and evolution of vertebrates. We have isolated a cDNA encoding an amphioxus (Branchiostoma belcheri) glutathione-S transferase with a predicted molecular mass of approximately 26 kDa, from the gut cDNA library. The glutathione-S-transferase had 43.7-51.8% identity to most glutathione-S-transferases identified from aquatic organisms including fish and green alga, but it was much less identical (<27%) to other cytosolic glutathione S-transferase classes. The phylogenetic analysis revealed that the glutathione-S transferase was grouped together with most piscine and algal glutathione-S transferases, separating from other cytosolic glutathione-S-transferase classes. Moreover, the glutathione-S-transferase had an exon-intron organization typical of zebrafish putative GST, red sea bream GSTR1 and plaice GSTA1 genes. The recombinant glutathione-S-transferase has been successfully expressed and purified, which showed a relatively high catalytic activity (3.37+/-0.1 unit/mg) toward 1-chloro-2, 4-dinitrobenzene and a moderate activity toward ethacrynic acid (0.41+/-0.01 unit/mg), although it had no detectable activity toward 1, 2 dichloro-4-nitrobenzene, 4-hydroxynonenal, 4-nitrobenzyl chloride and cumene hydroperoxide. In addition, we have revealed a tissue-specific expression pattern of the glutathione-S-transferase gene in B. belcheri, with the most abundant expression in the hepatic caecum. All these indicate that the amphioxus glutathione-S-transferase belongs to a novel rho-class of glutathione-S transferases with a tissue-specific expression pattern. The relation between the glutathione-S-transferase expression in amphioxus hepatic caecum and the origin of vertebrate liver is also discussed. PMID- 17084658 TI - Current concepts: contact lens related Pseudomonas keratitis. AB - Despite the development of silicone hydrogel lenses, Pseudomonas aeruginosa (PA) continues to be the leading cause of contact lens related microbial keratitis. Understanding the pathogenesis of PA-mediated corneal infection is critical to the development of new prevention and treatment strategies. Recently intracellular invasion of surface corneal epithelial cells by PA has been revisited as an important element in the infection process. This review identifies the mechanisms involved, and examines the roles of the lens, hypoxia alone, PA stain, cystic fibrosis transmembrane receptor protein (CFTR), and membrane lipid rafts in mediating intracellular invasion in both in vitro and in vivo conditions. Non-toxic blockade of raft formation in vitro or in vivo effectively abrogates PA internalization and may represent a unique, new strategy to prevent or ameliorate lens-related PA microbial keratitis. PMID- 17084659 TI - Biosynthesis of gold nanoparticles using Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa were used for extra-cellular biosynthesis of gold nanoparticles (Au NPs). Consequently, Au NPs were formed due to reduction of gold ion by bacterial cell supernatant of P. aeruginosa ATCC 90271, P. aeruginosa (2) and P. aeruginosa (1). The UV-vis and fluorescence spectra of the bacterial as well as chemical prepared Au NPs were recorded. Transmission electron microscopy (TEM) micrograph showed the formation of well-dispersed gold nanoparticles in the range of 15-30 nm. The process of reduction being extra-cellular and may lead to the development of an easy bioprocess for synthesis of Au NPs. PMID- 17084660 TI - Theoretical investigations of the specific heat functions for the orthorhombic Nd+3 centers in some crystals. AB - Garnets doped with Nd+3 have attracted extensive interests because of their applications in solid-state lasers, as well as upconversion luminescence and magneto-optical and magnetic properties. Usually, these properties are closely related to the local structures and properties of the Nd+3 impurities in the hosts. Since information about electronic states and local structures of the impurity Nd+3 in the garnets would be helpful to the understanding of the properties of these materials, theoretical investigations on the specific heat functions and the local structures for these Nd+3 centers are of significance. PMID- 17084661 TI - Optical absorption of methoxy and carboethoxy derivatives of 1,3-diphenyl-1H pyrazolo[3,4-b]quinoline. AB - Paper deals with experimental investigations and quantum chemical calculations of the optical absorption spectra of methoxy and carboethoxy 1,3-diphenyl derivatives of the pyrazoloquinoline ([PQ]): 6-methoxy-1,3-dyphenil-[PQ], 6 methoxy-1,3-(p-methoxyphenyl)-[PQ], 6-methoxy-1-(p-methoxyphenyl)-[PQ] and 6 carboethoxy-1,3-diphenyl-[PQ]. The quantum chemical calculations are performed by means of the semiempirical quantum chemical methods (AM1 or PM3) applied to: (a) the equilibrium molecular conformation in vacuo (T=0 K); (b) the molecular dynamic (MD) trajectory (T=300 K) which includes the dynamics of a certain molecular fragment (moiety) only (fragmental MD simulations); or (c) the MD trajectory obtained for most general case within the total MD simulations at T=300 K. The results of these calculations are compared with the measured spectra of the optical absorption. The quantum chemical simulations show that the dynamics of the methoxy or carboethoxy groups practically does not influence the absorption spectrum whereas the strongest its modification (3001:80), determined by constant potential amperometry at 830 mV (vs. Ag/AgCl). Selectivity for ascorbate and dopamine as well as detection limit for NO (1.5 nM at S/N=3) is 5-10 times better than parameters usually reported for Nafion coated porphyrinic sensors. Nafion coating can further enhance selectivity properties as well as aids to the stability of the sensors' responses. PMID- 17084680 TI - Validation of XP-C pathogenic variations in archival material from a live XP patient. AB - Xeroderma pigmentosum (XP) genetic complementation group C (XP-C) is the most common form of the disease worldwide. Thirty-four distinct genetic defects have been identified in 45 XP-C patients. Further identification of such defects and the frequency of their occurrence offers the potential of generating diagnostic and prognostic molecular screening panels. Archival material (such as formalin fixed paraffin embedded skin) may be useful for the identification of novel genetic variations and for documenting the frequency of individual genetic defects in patients who are no longer available for study. However, the use of archival material precludes direct analysis of changes in the mRNA resulting from genomic changes. The serendipitous reacquisition of an XP individual in whom genetic defects were previously characterized in archival material allowed confirmation of the defects as well as a direct analysis of the consequences of these defects on mRNA, mRNA expression and on cellular phenotypes. PMID- 17084681 TI - A HPLC-mass spectrometric method suitable for the therapeutic drug monitoring of everolimus. AB - We report here the validation of an HPLC-electrospray-tandem mass spectrometry method for the quantification of everolimus, an immunosuppressant drug. Whole blood samples (100 microl) were extracted by protein precipitation which involved sample pre-treatment with zinc sulphate followed by acetonitrile (containing internal standard, 40-O-(3'-hydroxy)propyl-rapamycin). HPLC was performed using a step-gradient at a flow rate of 0.6 ml/min on a Waters TDM C18 column (10 mm x 2.1mm I.D.) with a resultant chromatographic analysis time of 2 min. Mass spectrometric detection by selected reaction monitoring (everolimus m/z 975.5- >908.3; internal standard m/z 989.5-->922.3). The assay was linear from 0.5 to 40 microg/l (r2>0.994, n=11). The inter- and intra-day analytical recovery and imprecision for quality control samples (1.25, 12.5 and 30 microg/l) were 93.4 98.2% and <10.7%, respectively (n=10). At the lower limit of quantification (0.5 microg/l) the inter- and intra-day analytical recovery was 94.4-95.8% with imprecision of <14.1% (n=10). The absolute recovery of everolimus (6.5 microg/l) and internal standard (12.5 microg/l) was 96.5 and 88.3%, respectively (n=3). A comparison of our method against the mean of all HPLC methods for a series of samples from an external proficiency testing scheme revealed good correlation as shown by the regression analysis: y=0.973x+0.301 (r2=0.986, n=71). In conclusion, the method described is suited to the current requirements for therapeutic drug monitoring of everolimus. PMID- 17084682 TI - Congenital amusia. PMID- 17084683 TI - Alzheimer's discovery. PMID- 17084684 TI - Evolution of the neocortex. PMID- 17084685 TI - Chronic jet-lag increases mortality in aged mice. PMID- 17084686 TI - The importance of procedural knowledge in desert-ant navigation. PMID- 17084687 TI - Developmental biology: holding pattern for histones. AB - New research on lipid droplets in Drosophila embryos has led to the surprising conclusion that these poorly understood organelles have a novel role as a regulated storage depot of maternally supplied proteins, particularly histones. PMID- 17084688 TI - Homing behavior: decisions, dominance and democracy. AB - When flying in pairs, pigeons with different preferred paths back to the loft often agree on a joint route, and in doing so get home faster than either would have done on its own. PMID- 17084689 TI - Speech production: the force of your words. AB - Research on speech production has traditionally focused on how acoustic goals are met. A recent study shows that talking also involves somatosensory goals that do not have any acoustic consequences. PMID- 17084690 TI - Meiotic spindle: sculpted by severing. AB - Katanin is a conserved AAA ATPase with the ability to sever microtubules, but its biological function in animal cells has been obscure. A recent study using electron tomography has found that katanin stimulates the production of microtubules in the meiotic spindles of Caenorhabditis elegans oocytes. PMID- 17084691 TI - Spider silk: thousands of nano-filaments and dollops of sticky glue. AB - Some spiders use glues while others deploy strands of fine filaments for fixing flies. Recent work has provided new insights into the mechanical properties of these nano-scale ropes. PMID- 17084692 TI - Bacterial flagellum: visualizing the complete machine in situ. AB - Electron tomography of frozen-hydrated bacteria, combined with single particle averaging, has produced stunning images of the intact bacterial flagellum, revealing features of the rotor, stator and export apparatus. PMID- 17084693 TI - Language: specifying the site of modality-independent meaning. AB - Language processing can be triggered by auditory, visual or somatosensory input. A recent study has provided new insight into a fundamental issue raised by this observation: how is knowledge of language implemented in the human brain such that speakers can use any type of sensory-motor input-output system for comprehension and production? PMID- 17084694 TI - DNA-damage control: Claspin destruction turns off the checkpoint. AB - The ATR-Claspin-Chk1 pathway is critical for turning on the cellular response to DNA damage and replication stress. Five recent reports uncover new mechanisms controlling the recovery phase of the checkpoint response, and introduce crucial roles for Claspin, Rad17 phosphorylation and the ubiquitin proteasome pathway in Chk1 signaling. PMID- 17084695 TI - Spindle oscillations during asymmetric cell division require a threshold number of active cortical force generators. AB - BACKGROUND: Asymmetric division of the C. elegans zygote is due to the posterior directed movement of the mitotic spindle during metaphase and anaphase. During this movement along the anterior-posterior axis, the spindle oscillates transversely. These motions are thought to be driven by a force-generating complex-possibly containing the motor protein cytoplasmic dynein-that is located at the cell cortex and pulls on microtubules growing out from the spindle poles. A theoretical analysis indicates that the oscillations might arise from mechanical coordination of the force-generating motors, and this coordination is mediated by the load dependence of the motors' detachment from the microtubules. The model predicts that the motor activity must exceed a threshold for oscillations to occur. RESULTS: We have tested the existence of a threshold by using RNA interference to gradually reduce the levels of dynein light intermediate chain as well as GPR-1 and GPR-2 that are involved in the G protein mediated regulation of the force generators. We found an abrupt cessation of oscillations as expected if the motor activity dropped below a threshold. Furthermore, we can account for the complex choreography of the mitotic spindle the precise temporal coordination of the buildup and die-down of the transverse oscillations with the posterior displacement-by a gradual increase in the processivity of a single type of motor machinery during metaphase and anaphase. CONCLUSIONS: The agreement between our results and modeling suggests that the force generators themselves have the intrinsic capability of generating oscillations when opposing forces exceed a threshold. PMID- 17084696 TI - From compromise to leadership in pigeon homing. AB - A central problem faced by animals traveling in groups is how navigational decisions by group members are integrated, especially when members cannot assess which individuals are best informed or have conflicting information or interests . Pigeons are now known to recapitulate faithfully their individually distinct habitual routes home , and this provides a novel paradigm for investigating collective decisions during flight under varying levels of interindividual conflict. Using high-precision GPS tracking of pairs of pigeons, we found that if conflict between two birds' directional preferences was small, individuals averaged their routes, whereas if conflict rose over a critical threshold, either the pair split or one of the birds became the leader. Modeling such paired decision-making showed that both outcomes-compromise and leadership-could emerge from the same set of simple behavioral rules. Pairs also navigated more efficiently than did the individuals of which they were composed, even though leadership was not necessarily assumed by the more efficient bird. In the context of mass migration of birds and other animals, our results imply that simple self organizing rules can produce behaviors that improve accuracy in decision-making and thus benefit individuals traveling in groups . PMID- 17084697 TI - Involvement of human basal ganglia in offline feedback control of voluntary movement. AB - Practice makes perfect, but the neural substrates of trial-to-trial learning in motor tasks remain unclear. There is some evidence that the basal ganglia process feedback-related information to modify learning in essentially cognitive tasks , but the evidence that these key motor structures are involved in offline feedback related improvement of performance in motor tasks is paradoxically limited. Lesion studies in adult zebra finches suggest that the avian basal ganglia are involved in the transmission or production of an error signal during song . However, patients with Huntington's disease, in which there is prominent basal ganglia dysfunction, are not impaired in error-dependent modulation of future trial performance . By directly recording from the subthalamic nucleus in patients with Parkinson's disease, we demonstrate that this nucleus processes error in trial performance at short latency. Local evoked activity is greatest in response to smallest errors and influences the programming of subsequent movements. Accordingly, motor parameters are least likely to change after the greatest evoked responses so that accurately performed trials tend to precede other accurate trials. This relationship is disrupted by electrical stimulation of the nucleus at high frequency. Thus, the human subthalamic nucleus is involved in feedback-based learning. PMID- 17084698 TI - Differential regulation of germline mRNAs in soma and germ cells by zebrafish miR 430. AB - Early in development, primordial germ cells (PGCs) are set aside from somatic cells and acquire a unique gene-expression program . The mechanisms underlying germline-specific gene expression are largely unknown. Nanos expression is required during germline development and is posttranscriptionally restricted to PGCs . Here we report that the microRNA miR-430 targets the 3' untranslated region (UTR) of nanos1 during zebrafish embryogenesis. A miR-430 target site within the nanos1 3' UTR reduces poly(A) tail length, mRNA stability, and translation. Repression is disrupted in maternal-zygotic dicer mutants (MZdicer), which lack mature miRNAs , and is restored by injection of processed miR-430. Although miR-430 represses other genes equally in germline and soma, specific regions in the nanos1 3' UTR compensate for microRNA-mediated repression in PGCs and allow germline-specific expression. We show that the 3' UTR of an additional PGC-specific gene, TDRD7, is also targeted by miR-430. These results indicate that miR-430 targets the 3' UTRs of germline genes and suggest that differential susceptibility to microRNAs contributes to tissue-specific gene expression. PMID- 17084699 TI - Vectorial information for Arabidopsis planar polarity is mediated by combined AUX1, EIN2, and GNOM activity. AB - Cell polarity is commonly coordinated within the plane of a single tissue layer (planar polarity), and hair positioning has been exploited as a simple marker for planar polarization of animal epithelia . The root epidermis of the plant Arabidopsis similarly reveals planar polarity of hair localization close to root tip-oriented (basal) ends of hair-forming cells . Hair position is directed toward a concentration maximum of the hormone auxin in the root tip , but mechanisms driving this plant-specific planar polarity remain elusive. Here, we report that combinatorial action of the auxin influx carrier AUX1, ETHYLENE INSENSITIVE2 (EIN2) , and GNOM genes mediates the vector for coordinate hair positioning. In aux1;ein2;gnom eb triple mutant roots, hairs display axial (apical or basal) instead of coordinate polar (basal) position, and recruitment of Rho-of-Plant (ROP) GTPases to the hair initiation site reveals the same polar to-axial switch. The auxin concentration gradient is virtually abolished in aux1;ein2;gnom eb roots, where locally applied auxin can coordinate hair positioning. Moreover, auxin overproduction in sectors of wild-type roots enhances planar ROP and hair polarity over long and short distances. Hence, auxin may provide vectorial information for planar polarity that requires combinatorial AUX1, EIN2, and GNOM activity upstream of ROP positioning. PMID- 17084700 TI - An entropic mechanism to generate highly cooperative and specific binding from protein phosphorylations. AB - Cooperative interactions are essential to the operation of many biochemical networks. Such networks then respond ultrasensitively in a nonlinear manner to linear changes in network input, and network output, for example, levels of a phosphorylated protein or of gene expression, becomes a sigmoidal function of concentrations of input molecules. We present a novel, entropic ultrasensitivity mechanism that generates highly cooperative and specific binding between two proteins. We consider a disordered protein with multiple phosphorylation sites that binds to a single binding site on an interacting protein. We assume that each phosphorylation locally orders the protein. Such local order affects protein conformational entropy nonlinearly and generates binding that is a highly cooperative function of the number of protein phosphorylations (with Hill coefficients well above 10). Substantial binding may only occur once the disordered protein is phosphorylated a critical number of times or more. Cooperativity is determined by the size of the disordered region of the protein, the binding affinity, and unusually the concentration of the interacting protein. Given the widespread occurrence of disordered, multiply phosphorylated proteins, its highly ultrasensitive character, and the ease of its control, entropic, phosphorylation-driven cooperativity may be extensively exploited intracellularly. PMID- 17084701 TI - Expulsion of live pathogenic yeast by macrophages. AB - Phagocytic cells, such as neutrophils and macrophages, perform a critical role in protecting organisms from infection by engulfing and destroying invading microbes . Although some bacteria and fungi have evolved strategies to survive within a phagocyte after uptake, most of these pathogens must eventually kill the host cell if they are to escape and infect other tissues . However, we now demonstrate that the human fungal pathogen Cryptococcus neoformans is able to escape from within macrophages without killing the host cell by a novel expulsive mechanism. This process occurs in both murine J774 cells and primary human macrophages. It is extremely rapid and yet can occur many hours after phagocytosis of the pathogen. Expulsion occurs independently of the initial route of phagocytic uptake and does not require phagosome maturation . After the expulsive event, both the host macrophage and the expelled C. neoformans appear morphologically normal and continue to proliferate, suggesting that this process may represent an important mechanism by which pathogens are able to escape from phagocytic cells without triggering host cell death and thus inflammation . PMID- 17084702 TI - Phagosome extrusion and host-cell survival after Cryptococcus neoformans phagocytosis by macrophages. AB - Cryptococcus neoformans (Cn) is an encapsulated yeast that is a facultative intracellular pathogen and a frequent cause of human disease. The interaction of Cn with alveolar macrophages is critical for containing the infection , but Cn can also replicate intracellularly and lyse macrophages . Cn has a unique intracellular pathogenic strategy that involves cytoplasmic accumulation of polysaccharide-containing vesicles and intracellular replication leading to the formation of spacious phagosomes in which multiple cryptococcal cells are present . The Cn intracellular pathogenic strategy in macrophages and amoebas is similar, leading to the proposal that it originated as a mechanism for survival against phagocytic predators in the environment . Here, we report that under certain conditions, including phagosomal maturation, possible actin depolymerization, and homotypic phagosome fusion, Cn can exit the macrophage host through an extrusion of the phagosome, while both the released pathogen and host remain alive and able to propagate. The phenomenon of "phagosomal extrusion" indicates the existence of a previously unrecognized mechanism whereby a fungal pathogen can escape the intracellular confines of mammalian macrophages to continue propagation and, possibly, dissemination. PMID- 17084703 TI - Microtubule acetylation promotes kinesin-1 binding and transport. AB - Long-distance intracellular delivery is driven by kinesin and dynein motor proteins that ferry cargoes along microtubule tracks . Current models postulate that directional trafficking is governed by known biophysical properties of these motors-kinesins generally move to the plus ends of microtubules in the cell periphery, whereas cytoplasmic dynein moves to the minus ends in the cell center. However, these models are insufficient to explain how polarized protein trafficking to subcellular domains is accomplished. We show that the kinesin-1 cargo protein JNK-interacting protein 1 (JIP1) is localized to only a subset of neurites in cultured neuronal cells. The mechanism of polarized trafficking appears to involve the preferential recognition of microtubules containing specific posttranslational modifications (PTMs) by the kinesin-1 motor domain. Using a genetic approach to eliminate specific PTMs, we show that the loss of a single modification, alpha-tubulin acetylation at Lys-40, influences the binding and motility of kinesin-1 in vitro. In addition, pharmacological treatments that increase microtubule acetylation cause a redirection of kinesin-1 transport of JIP1 to nearly all neurite tips in vivo. These results suggest that microtubule PTMs are important markers of distinct microtubule populations and that they act to control motor-protein trafficking. PMID- 17084704 TI - Regulation of ras signaling dynamics by Sos-mediated positive feedback. AB - The RTK-Ras-ERK cascade is a central signaling module implicated in the control of diverse biological processes including cell proliferation, differentiation, and survival. The coupling of RTK to Ras is mediated by the Ras-specific nucleotide-exchange factor Son of Sevenless (Sos), which activates Ras by inducing the exchange of GDP for GTP . Considerable evidence indicates that the duration and amplitude of Ras signals are important determinants in controlling the biological outcome . However, the mechanisms that regulate the quantitative output of Ras signaling remain poorly understood. We define a previously unrecognized regulatory component of the machinery that specifies the kinetic properties of signals propagated through the RTK-Ras-ERK cascade. We demonstrate that the establishment of a positive feedback loop involving Ras.GTP and Sos leads to an increase in the amplitude and duration of Ras activation in response to EGF stimulation. This effect is propagated to downstream elements of the pathway as reflected by sustained EGF-induced ERK phosphorylation and enhanced SRE-dependent transcription. As a consequence, the physiological endpoint of EGF action is switched from proliferation to differentiation. We propose that the engagement of Ras/Sos positive feedback loop may contribute to the mechanism by which ligand stimulation is coupled to discrete biological responses. PMID- 17084705 TI - EXtENDINg beta cell survival by UPRegulating ATF4 translation. AB - In this issue of Cell Metabolism, Daniel Drucker and colleagues (Yusta et al., 2006) explore how the incretin mimetic exendin-4 improves beta cell function and survival during ER stress. Their findings suggest that protein kinase A signaling elicited by GLP-1 receptor activation differentially modulates one arm of the unfolded protein response (UPR). Regulation of this UPR pathway leads to enhanced translational expression of ATF4, a transcription factor central for stress remedy and cell survival. PMID- 17084706 TI - Life and death of the distal nephron: WNK4 and NCC as major players. AB - Missense mutations in the WNK4 gene lead to the development of familial hyperkalemic hypertension, a rare form of human hypertension. It was shown in vitro that WNK4 regulates the surface expression and activity of a number of ion channels and transporters. The in vivo analysis of wild-type and mutant WNK4 overexpression in transgenic mice models demonstrated that this serine-threonine kinase controls ion handling in the kidney mainly, and probably exclusively, through the regulation of the Na-Cl contransporter NCC activity. PMID- 17084707 TI - Banking on ATM as a new target in metabolic syndrome. AB - In this issue of Cell Metabolism, Semenkovich and his colleagues show that ATM, a protein well known for its roles in the cellular response to DNA breaks, may also be linked to metabolic and cardiovascular diseases (Schneider et al., 2006). ATM seemingly does this by inhibiting JNK, a stress kinase involved in inflammation with related effects in insulin resistance and atherosclerosis. In an interesting twist, the authors show that chloroquine, an antimalarial drug, also activates ATM, which inhibits JNK, and improves insulin sensitivity and cardiovascular effects. These findings provide potential new insights into the pathogenesis and treatment of metabolic syndrome. PMID- 17084708 TI - Discovering orphans' sweet secret: NR4A receptors and hepatic glucose production. AB - Aberrant hepatic gluconeogenesis contributes importantly to hyperglycemia in type II diabetic patients. A study by Pei et al. (2006b) identifies NR4A orphan nuclear receptors as a novel branch of cAMP-dependent regulators of hepatic glucose production under healthy and diabetic conditions. PMID- 17084709 TI - Convergence between bone and energy homeostases: leptin regulation of bone mass. AB - The observation that obesity protects from osteoporosis suggested that energy metabolism and bone mass could be regulated by the same hormones. Testing this hypothesis revealed that leptin regulates bone mass through a hypothalamic relay and using two neural mediators, the sympathetic tone and CART, both acting on one cell type the osteoblast. This review summarizes the genetic and molecular bases of this regulation and discusses its potential clinical implications. PMID- 17084710 TI - Beneficial metabolic effects of M3 muscarinic acetylcholine receptor deficiency. AB - Most animal models of obesity and hyperinsulinemia are associated with increased vagal cholinergic activity. The M3 muscarinic acetylcholine receptor subtype is widely expressed in the brain and peripheral tissues and plays a key role in mediating the physiological effects of vagal activation. Here, we tested the hypothesis that the absence of M3 receptors in mice might protect against various forms of experimentally or genetically induced obesity and obesity-associated metabolic deficits. In all cases, the lack of M3 receptors greatly ameliorated impairments in glucose homeostasis and insulin sensitivity but had less robust effects on overall adiposity. Under all experimental conditions tested, M3 receptor-deficient mice showed a significant elevation in basal and total energy expenditure, most likely due to enhanced central sympathetic outflow and increased rate of fatty-acid oxidation. These findings suggest that the M3 receptor may represent a potential pharmacologic target for the treatment of obesity and associated metabolic disorders. PMID- 17084711 TI - ATM-dependent suppression of stress signaling reduces vascular disease in metabolic syndrome. AB - Metabolic syndrome is associated with insulin resistance and atherosclerosis. Here, we show that deficiency of one or two alleles of ATM, the protein mutated in the cancer-prone disease ataxia telangiectasia, worsens features of the metabolic syndrome, increases insulin resistance, and accelerates atherosclerosis in apoE-/- mice. Transplantation with ATM-/- as compared to ATM+/+ bone marrow increased vascular disease. Jun N-terminal kinase (JNK) activity was increased in ATM-deficient cells. Treatment of ATM+/+apoE-/- mice with low-dose chloroquine, an ATM activator, decreased atherosclerosis. In an ATM-dependent manner, chloroquine decreased macrophage JNK activity, decreased macrophage lipoprotein lipase activity (a proatherogenic consequence of JNK activation), decreased blood pressure, and improved glucose tolerance. Chloroquine also improved metabolic abnormalities in ob/ob and db/db mice. These results suggest that ATM-dependent stress pathways mediate susceptibility to the metabolic syndrome and that chloroquine or related agents promoting ATM activity could modulate insulin resistance and decrease vascular disease. PMID- 17084712 TI - GLP-1 receptor activation improves beta cell function and survival following induction of endoplasmic reticulum stress. AB - Perturbation of endoplasmic reticulum (ER) homeostasis impairs insulin biosynthesis, beta cell survival, and glucose homeostasis. We show that a murine model of diabetes is associated with the development of ER stress in beta cells and that treatment with the GLP-1R agonist exendin-4 significantly reduced biochemical markers of islet ER stress in vivo. Exendin-4 attenuated translational downregulation of insulin and improved cell survival in purified rat beta cells and in INS-1 cells following induction of ER stress in vitro. GLP 1R agonists significantly potentiated the induction of ATF-4 by ER stress and accelerated recovery from ER stress-mediated translational repression in INS-1 beta cells in a PKA-dependent manner. The effects of exendin-4 on the induction of ATF-4 were mediated via enhancement of ER stress-stimulated ATF-4 translation. Moreover, exendin-4 reduced ER stress-associated beta cell death in a PKA dependent manner. These findings demonstrate that GLP-1R signaling directly modulates the ER stress response leading to promotion of beta cell adaptation and survival. PMID- 17084713 TI - PGC1alpha expression is controlled in skeletal muscles by PPARbeta, whose ablation results in fiber-type switching, obesity, and type 2 diabetes. AB - Mice in which peroxisome proliferator-activated receptor beta (PPARbeta) is selectively ablated in skeletal muscle myocytes were generated to elucidate the role played by PPARbeta signaling in these myocytes. These somatic mutant mice exhibited a muscle fiber-type switching toward lower oxidative capacity that preceded the development of obesity and diabetes, thus demonstrating that PPARbeta is instrumental in myocytes to the maintenance of oxidative fibers and that fiber-type switching is likely to be the cause and not the consequence of these metabolic disorders. We also show that PPARbeta stimulates in myocytes the expression of PGC1alpha, a coactivator of various transcription factors, known to play an important role in slow muscle fiber formation. Moreover, as the PGC1alpha promoter contains a PPAR response element, the effect of PPARbeta on the formation and/or maintenance of slow muscle fibers can be ascribed, at least in part, to a stimulation of PGC1alpha expression at the transcriptional level. PMID- 17084714 TI - RNA interference: mechanisms of action and therapeutic consideration. PMID- 17084715 TI - International integrated database for the evaluation of severe sepsis and drotrecogin alfa (activated) therapy: analysis of efficacy and safety data in a large surgical cohort. AB - BACKGROUND: The International Integrated Database for the Evaluation of Severe Sepsis and Drotrecogin alfa (activated) Therapy includes an extensive cohort of surgical patients (1659/4459; 37%). This database broadens the experience reported on a comparatively small set of surgical patients from the pivotal Protein C Worldwide Evaluation in Severe Sepsis trial to examine issues of safety and efficacy in a much larger cohort. METHODS: We conducted a retrospective analysis of prospectively defined outcomes from 5 integrated clinical studies of severe sepsis. Multivariable analyses incorporated propensity scores, treatment, and significant baseline risk factors as independent variables in logistic regression models for 2 outcomes: serious adverse events that were observed during infusion and 28-day, all-cause mortality rates. Adjusted odds ratios were calculated for clinically important strata. Multiple subcategories of serious bleeding-event rates are presented. RESULTS: Although surgical patients who were treated with drotrecogin alfa [activated] (DrotAA) experienced a greater proportion of serious bleeding events during the infusion period, most of the patients were treated without fatal consequence. A 10.7% absolute all cause mortality risk reduction (adjusted odds ratio, 0.66; 95% CI, 0.45-0.97) was observed for DrotAA-treated, high-risk (Acute Physiology and Chronic Health Evaluation II, >or=25) surgical patients. We could not demonstrate a survival benefit in DrotAA-treated, low-risk (Acute Physiology and Chronic Health Evaluation II, <25) surgical patients. When surgical patients were stratified by number of organ dysfunctions, absolute risk reductions were observed in both categories: multiorgan (4.3%) and single (4.5%). CONCLUSION: International Integrated Database for the Evaluation of Severe Sepsis and Drotrecogin alfa (activated) Therapy analyses affirmed the favorable benefit/risk profile of DrotAA for surgical patients. The serious adverse event rate that was experienced by surgical patients during the study drug infusion period was 7.5% in the DrotAA treated group versus 6.3% in the placebo-treated group (odds ratio, 1.41; 95% CI, 0.89-2.25). The clinical benefit of DrotAA therapy paralleled baseline risk of death and substantiated findings from the Protein C Worldwide Evaluation in Severe Sepsis study. Future analyses are needed to evaluate the special relationships among sepsis severity, bleeding management, and the postoperative timing of DrotAA administration. PMID- 17084716 TI - Informed consent, capitation, and conflicts of interest in clinical trials: views from the field. AB - BACKGROUND: The beliefs of research team members about informed consent practices are not well studied, especially in regard to financial aspects of research such as capitation fees. Sponsors of research offer capitation fees for the enrollment of subjects, and although this money is intended to cover the costs of the research it can sometimes be used in other ways. There is no clear consensus about whether this financial aspect of trials should be part of the informed consent process or whether it represents a potential conflict of interest. METHODS: We presented several hypothetical cases to members of the American College of Surgeons Oncology Group (ACOSOG) at a semi-annual meeting. ACOSOG is a federally funded clinical trials group evaluating surgical and adjuvant methods of cancer treatment. The meeting included surgeons, nurses, case managers, and others involved in clinical surgical trials. RESULTS: We found that most respondents believed that it would be a violation of the protocol to enroll a subject who showed lack of recall about the study. Most respondents also favored disclosure of capitation fees, especially if those fees went into discretionary accounts, but there was some disagreement about the benefit of that disclosure to potential participants. CONCLUSIONS: This study shows the need for greater work in ensuring that research team members share common understandings about informed consent. It also shows that most of these research team participants are prepared to make more disclosure about financial aspects of research than current standards in fact require. PMID- 17084717 TI - Small remnant liver volume after right lobe living donor hepatectomy. AB - BACKGROUND: Right lobe living donor liver transplantation has become a viable option for adult patients with end-stage liver disease, however, the safety of the donor is of paramount importance. One of the key factors in donor safety is ensuring adequate donor remnant liver volume. METHODS: We retrospectively examined donors who had less than 30% remnant liver volume after right graft procurement. Eighty-six right lobe living donor transplants were carried out in Chang Gung Memorial Hospital, Kaohsiung Medical Center, from January 1999 to December 2004. RESULTS: Eight donors had less than 30% remnant liver volume (Group 1) after graft procurement and 78 donors had remnant liver volume greater than 30% (Group 2). There were no differences in donor characteristics, types of graft, operative parameters, and post-operative liver and renal function as well as liver volume at 6 months post-donation between the 2 groups. The graft weight obtained in Group 1 donors was significantly greater compared with that from Group 2 (P<.005). The overall donor complication rate was 6.98%, and all the complications occurred among group 2 donors. CONCLUSIONS: The judicious use of donors with less than 30% remnant liver volume is safe as a last resort. PMID- 17084718 TI - Differential diagnosis of proximal biliary obstruction. AB - BACKGROUND: Obstruction at the hepatic duct confluence is generally due to hilar cholangiocarcinoma (HCCA). However, in up to 15% of patients, hilar obstruction could be due to alternative diagnoses other than HCCA. The aim of this study was to determine preoperative criteria that could differentiate HCCA from the alternative diagnoses. METHODS: All patients with hilar obstruction presumed to represent HCCA were included (1997-2001). The extent of disease was assessed preoperatively with computed tomography, magnetic resonance cholangiopancreatography, and Duplex ultrasonography, and these findings were correlated to the final histopathology. RESULTS: A total of 171 patients were included in the study, with HCCA being the most common diagnosis (141 patients [82.4%], group I). Alternative diagnoses other than HCCA were encountered in 30 patients (17.5%, group II) and included benign stricture (9 patients [5.2%]) and other malignancy (21 patients [12%]). There was a higher incidence of involvement of the second-order bile ducts in group I (26% vs 3% in group II, P<.01). Vascular involvement and lobar atrophy were more common in group I (58% and 41%) when compared with group II (16% and 6%, P<.005 and P<.002). The combination of these 2 findings (vascular invasion+lobar atrophy) was reliable for discriminating patients with HCCA from the alternative diagnoses. (38% in group I and 3.3% in group II, P<.001). CONCLUSIONS: Involvement of second-order bile ducts, vascular invasion, and lobar atrophy are more likely in patients with HCCA. The combination of vascular invasion and lobar atrophy significantly increases the diagnostic likelihood of HCCA. The absence of these findings should raise awareness of the possibility of an alternative diagnosis. PMID- 17084719 TI - Resected periampullary adenocarcinoma: 5-year survivors and their 6- to 10-year follow-up. AB - BACKGROUND: Many studies have reported 5-year survival data after pancreaticoduodenectomy for periampullary adenocarcinoma. This study evaluates 10 year survival in patients surviving 5 years after initial surgery. METHODS: We reviewed all patients undergoing pancreaticoduodenectomy for periampullary adenocarcinoma from April 1970 to July 1999 at a single institution. All 5-year survivors were identified, and their subsequent 5-year survival was compared with the actuarial survival of the general population starting at 70 years of age. RESULTS: Nine hundred fifteen patients underwent pancreaticoduodenectomy for periampullary adenocarcinoma. Follow-up was complete on 890 patients. There were 201 (23%) 5-year survivors with a median age of 65 years at initial surgery; 51% were male and 92% were Caucasian. For the 5-year survivors, the carcinoma origin was pancreatic in 46%, ampullary in 25%, distal bile duct in 17%, and duodenal in 12%. For all 5-year survivors, the subsequent 5-year actuarial survival rate was 65%, with a median survival after achieving the 5-year landmark of 7.9 additional years. The subsequent 5-year survival by site of tumor origin was 55% for pancreatic, 66% for ampullary, 74% for bile duct, and 85% for duodenal cancer. For the age-matched population, the 5-year survival rate was 87% (P<.001 when compared with those with all periampullary cancers). CONCLUSIONS: While the 5 year survival rate for all patients with resected periampullary adenocarcinoma is only 23%, these data imply that attainment of the 5-year survival landmark carries with it an improved survival for the subsequent 5 years. While the survival rate was less than that of the age-matched population, 65% of 5-year survivors survived 5 more years, bringing them to the 10-year postresection landmark. PMID- 17084720 TI - Evaluation and management of thoracopancreatic fistula. AB - BACKGROUND: Thoracopancreatic fistula is a rare clinical entity but a serious complication of inflammatory pancreatic diseases, caused by a disruption of the pancreatic ductal system. Its diagnosis is frequently misleading, however, and thus is often delayed. METHODS: Seven patients with thoracopancreatic fistula who presented at our department between March 2002 and July 2005 were investigated, focusing on the diagnostic work-up as well as the treatment strategies, the response to therapy, and the outcome. RESULTS: Thoracopancreatic fistulas developed secondary to alcohol-related chronic pancreatitis in 6 patients and acute severe pancreatitis in 1. The disruption sites of the pancreatic ductal system were the head of the pancreas in 2 patients, the pancreatic body in 2 patients, and the pancreatic tail in 3 patients. All patients, except 1, were complicated with stricture of the main pancreatic duct, with ductal disruptions developing distal to the pancreatic strictures. The precise demonstration of the pancreatic ductal anatomy with ultrasonography, computed tomography (CT), conventional magnetic resonance imaging (MRI), and endoscopic retrograde cholangiopancreatography was limited. In contrast, MR-cholangiopancreatography (MRCP) provided excellent mapping of the pancreatic ductal stricture, disruption, and fistula in 6 patients. Various medical therapies failed to close the fistula in all patients. Subsequent treatments, based on the assessment of pancreatic ductal anatomy with MRCP, included endoscopic transpapillary implantation of a pancreatic stent, a longitudinal pancreaticojejunostomy, distal pancreatectomy, and peritoneal drainage. All patient outcomes were favorable. CONCLUSIONS: MRCP is an essential diagnostic modality in all suspected cases of thoracopancreatic fistula. The goal of treatment should be directed toward a sufficient decompression of the obstructed pancreas. If severe pancreatic stricture is present, then surgical decompression may be required in accordance with the individual pancreatic ductal anatomy. PMID- 17084721 TI - Needle core biopsy in the diagnosis of phyllodes neoplasm. AB - BACKGROUND: Accurate identification of phyllodes neoplasms without surgical intervention is difficult, reducing the ability to manage "benign" lumps non operatively and impacting on the open benign biopsy rate. Needle core biopsy is considered to be a highly accurate technique in the diagnosis of breast carcinoma. Its accuracy in the diagnosis of phyllodes neoplasm has not been established. METHODS: A series of 3729 core biopsies performed between January 1999 and July 2005 were examined. All core biopsies followed by surgical excision were identified. Histologic concordance between core biopsy and excision specimen was analyzed. RESULTS: Twenty-three patients had phyllodes neoplasm on excisional biopsy with prior core biopsy findings as follows: phyllodes neoplasm (n=2), "equivocal" for phyllodes neoplasm (n=12), fibroadenoma (n=3), benign (n=6). The false negative rate for phyllodes neoplasm was therefore 39% (n=9/23). Of the total biopsy series, 35 patients had a core biopsy suggesting the possibility of phyllodes neoplasm. Of these, 32% (n=11) were found to be phyllodes neoplasm on excision, 3% (n=1) phyllodes neoplasm with breast carcinoma, 6% (n=2) breast carcinoma, and 3% (n=1) sarcoma. When a preference for phyllodes neoplasm (n=4) was stated on the equivocal core biopsies, excision correlated with the stated preference; this correlation also occurred in 90% (n=9/10) of core biopsies where fibroadenoma was favored. CONCLUSIONS: Needle core biopsy rarely produces a definite preoperative diagnosis of phyllodes neoplasm. A diagnosis of fibroadenoma or equivocal phyllodes neoplasm on core biopsy should not prevent excision if clinical suspicion remains. PMID- 17084722 TI - Radiolabeled and near-infrared fluorescent fibrinogen derivatives create a system for the identification and repair of obscure gastrointestinal bleeding. AB - BACKGROUND: Gastrointesintal hemorrhage is often difficult to localize. A test that does not depend on active bleeding might prove clinically useful. We tested 2 novel fibrinogen (FBG)-based contrast agents for their ability to localize gastrointestinal (GI) hemorrhage after bleeding stopped. 125I-FBG permits gamma ray-based preoperative or intraoperative scanning, and near-infrared (NIR) flourescent FBG (FBG800) permits real-time intraoperative visualization of active clot. METHODS: Bovine FBG was radiolabeled with 125I or conjugated to the NIR fluorophore CW800. Sites of bleeding were created by gastrotomy, mucosal resection of the stomach, or laceration of a mesenteric vessel; then 1.7 mg/kg FBG800 or 15 microCi/kg 125I-FBG was injected intravenously into mice, rabbits, or pigs 30 minutes before or after injury. Sites of active clot were quantified by using gamma counting and were also imaged by using invisible NIR light intraoperatively, for up to 3 hours postinjection. RESULTS: After an injection of either 125I-FBG or FBG800, sites of prior bleeding could be identified in the absence of active bleeding. Blood clearance was such that a signal-to-background ratio of 2.0 or greater could be achieved within 20 minutes after injection. A similarly labeled human serum albumin did not accumulate at any site, with an SBR of 1.0 or less. CONCLUSIONS: Both radiolabeled (preoperative gamma scanning) and NIR fluorescent (intraoperative real-time imaging) FBG can be used in experimental situations to identify the location of prior bleeding in the absence of active bleeding. Taken together, these contrast agents create a system for the identification and control of obscure GI bleeding. PMID- 17084723 TI - Interleukin-6 and STAT3 protect the liver from hepatic ischemia and reperfusion injury during ischemic preconditioning. AB - BACKGROUND: Ischemic preconditioning has been shown to protect the liver from ischemia/reperfusion injury. We hypothesized that IL-6 directly modulates the protective effects of ischemic preconditioning. METHODS: Three weeks after undergoing splenic transposition, wild-type C57BL/6 and IL-6 null mice underwent 75 minutes of total hepatic ischemia with or without prior ischemic preconditioning (10 minutes of ischemia followed by 15 minutes of reperfusion). After reperfusion, serum ALT, serum IL-6, hepatic IL-6 mRNA, hepatic pSTAT3, and liver histology were evaluated. RESULTS: In wild-type mice, survival at 24 hours was greater in the preconditioned group compared with the non-preconditioned group (75% vs 40%, P<.05). In IL-6 null mice, however, ischemic preconditioning did not improve survival when compared with the non-preconditioned group. Preconditioning significantly reduced hepatocellular injury in wild-type mice (P<.05) when compared with IL-6 null animals. This protection was associated with significant increases in serum IL-6, hepatic IL-6 mRNA, and hepatic pSTAT3 levels (P<.05). The protective effects of ischemic preconditioning that correlated with significant increases in systemic IL-6, hepatic IL-6 mRNA abundance, and pSTAT3 levels, were not observed in IL-6 null mice. CONCLUSIONS: The protective effects of ischemic preconditioning during total hepatic ischemia/reperfusion injury are dependent on IL-6 signaling and are associated with increased phosphorylation of hepatic STAT3. PMID- 17084724 TI - Pancreatic intraepithelial neoplasia and ductal adenocarcinoma induced by DMBA in mice. AB - BACKGROUND: Pancreatic ductal adenocarcinoma has a poor long-term prognosis. Experimental models are necessary to understand not only its biologic behavior, but also the early pancreatic lesions known as pancreatic intraepithelial neoplasia (PanIN) and to develop new treatments. The aim of this study was to evaluate pancreatic carcinogenesis induced by 7,12-dimethyl-1,2-benzanthracene (DMBA) implantation in mice according to the PanIN classification system. METHODS: Ninety male, Mus musculus, CF-1 mice underwent a median laparotomy and 1 mg of DMBA was implanted into the proximal pancreas held in place by a purse string suture. Mice were killed after 30 and 60 days after which the excised pancreata were fixed in formalin, embedded in paraffin, and stained with hematoxylin-eosin for histologic analysis. The specimens were evaluated blind by 2 pathologists for the presence of the following histology: normal ducts, reactive hyperplasia, PanIN-1A, PanIN-1B, PanIN-2, and PanIN-3, and adenocarcinoma. RESULTS: In the 30-day group, pathologic evaluation showed 4 (17%) reactive hyperplasia, 16 (67%) PanIN lesions, and 4 (17%) adenocarcinomas. In the 60-day group, there were 10 (27%) specimens with reactive hyperplasia, 13 (35%) with PanIN lesions, and 14 (38%) with adenocarcinomas. The difference between groups was statistically significant (P<.05). All pancreata with adenocarcinoma had concomitant PanIN lesions. CONCLUSIONS: The DMBA experimental model in mice induces PanIN lesions and ductal adenocarcinoma that have similar histology to that of human pancreatic cancer. This model may be useful for study of pancreatic carcinogenesis, particularly the molecular progression of early pancreatic ductal lesions. PMID- 17084725 TI - Amiloride moderates increased gut permeability and diminishes mesenteric lymph mediated priming of neutrophils in trauma/hemorrhagic shock. AB - BACKGROUND: Amiloride, an inhibitor of Na+/H+ exchangers and Na+ channels has been shown recently to ameliorate both gut and lung injury in rats subjected to a combined insult of trauma and hemorrhagic shock (T/HS). We have shown previously that mesenteric lymph duct ligation prevents T/HS-induced lung endothelial injury and neutrophil activation, suggesting that toxic inflammatory factors originating from the gut and carried in the lymph are responsible for the lung injury observed after T/HS. This study investigates whether the protective effect of amiloride against T/HS-induced lung injury was associated with decreased lymph toxicity and gut permeability. METHODS: Male rats subjected to trauma (laparotomy) plus hemorrhagic shock (mean arterial pressure, 30 mm Hgx90 min) (T/HS) or trauma plus sham shock (T/SS) and treated with amiloride or its vehicle had their mesenteric lymph duct catheterized. Mesenteric lymph collected before and after shock was assayed for biologic activity on endothelial cells (cytotoxicity and permeability) and neutrophils (respiratory burst activity). Gut permeability was assessed by monitoring plasma concentrations of the fluorescent dye FITC-dextran after its injection into the ileum. RESULTS: Amiloride administration reduced the capacity of post-shock mesenteric lymph to prime neutrophils for an increased respiratory burst. Amiloride failed to decrease the ability of mesenteric lymph to kill endothelial cells or increase their permeability. Amiloride decreased gut permeability. CONCLUSIONS: The mechanisms of the lung protective effect of amiloride in rats undergoing T/HS may be secondary to decreased neutrophil activation, diminished gut permeability, or an effect on the end organ. PMID- 17084726 TI - Familial adenomatous polyposis predisposes to pathologic exposure of the stomach to bilirubin. AB - BACKGROUND: The role of duodenogastric reflux in the genesis of gastric polyps in familial adenomatous polyposis (FAP), although suggested by scintigraphy scanning studies, remains unclear. METHODS: Twenty-four hour intragastric bilirubin monitoring with the Bilitec optoelectronic device was carried out in 25 FAP patients, of whom 19 had gastric polyps (fundic gland in 13, adenomatous in 2, and both histologic types in 4) on endoscopic examination. Gastric exposure to bilirubin was expressed as the percentage of total recording time that absorbance exceeded the threshold of 0.25 and was calculated in reference to values obtained from 25 healthy volunteers. Helicobacter pylori status of the stomach was checked as well. RESULTS: Gastric exposure to bilirubin was pathologic in 14 (56%) patients. Gastric exposure to bilirubin was of longer duration in FAP patients than in healthy volunteers (mean+/-SEM: 19%+/-4% vs 6%+/-2%) (P<.005). It increased from healthy volunteers (6%+/-2%) to FAP patients without gastric polyps (10%+/-3%), and to FAP patients with gastric polyps (22%+/-5%) (P<.004). Bilirubin exposure times were similar in FAP patients with fundic gland polyps only and in those having either adenomatous polyps only or both types of polyps (24%+/-7% vs 17%+/-4%). No patient with pathologic gastric exposure to bilirubin as well as none having gastric polyps, had H. pylori in the antrum. CONCLUSIONS: This study shows that gastric exposure to bilirubin is of longer duration in FAP patients than in healthy volunteers, and in FAP patients with gastric polyps than in those without polyps. This study supports the existence of a direct correlation between pathologic duodenogastric reflux (DGR), the absence of H. pylori in the antrum, and the presence of gastric polyps in FAP patients. PMID- 17084727 TI - Anatomical and technical aspects of hepatic artery reconstruction in living donor liver transplantation. AB - BACKGROUND: We describe our experience with arterial reconstruction in living donor liver transplantation (LDLT) focusing on anatomic and technical aspects. METHODS: From June 1994 to February 2003, 132 grafts were implanted in 130 LDLT recipients including 1 re-transplant and 1 dual graft transplantation. Donor and recipient records were retrospectively reviewed. Anatomical variations in graft arteries were classified as: Type I, single pedicle with (Ia) or without (Ib) aberrant artery (left hepatic artery (HA) from left gastric artery or right HA from superior mesenteric artery); Type II, double pedicles with (IIa) or without (IIb) aberrant artery; Type III, equal to or greater than 3 pedicles. Statistical analyses were carried out using Mann-Whitney U-test. RESULTS: There were 72 male and 58 female recipients. The median age at transplantation was 3 years (range, 0.5 to 61). In left grafts, there were 34 Type Ia, 6 Type Ib, 33 Type IIa, 13 Type IIb, and 3 Type III; whereas in right grafts, there were 35 Type Ia, 6 Type Ib, 1 Type IIa, and 1 Type IIb. Two-in-one (2-in-1) segmental resection technique in graft HA harvest was carried out whenever there were tiny arteries supplying the donor graft. All HA reconstructions were done under microvascular techniques. There was no donor mortality and 1 recipient in-hospital mortality. There was no graft or patient loss due to HA occlusion. Donor complications included 3 biloma, 1 bile leak, 1 biliary stricture, and 1 late intestinal obstruction secondary to postoperative adhesions that were all successfully managed by non-operative interventions, except the biliary stricture that needed a revision to Roux-en-Y hepatico-jejunostomy. The 1-year and 5-year recipient survivals were 98% and 94%, respectively. CONCLUSIONS: Successful HA reconstruction can be safely carried out in LDLT recipients and live donors with multiple graft arteries using the 2-in-1 segmental resection of donor HA under microvascular techniques. PMID- 17084729 TI - Notes from an Air Force Hospital in Iraq. PMID- 17084730 TI - Malignant gastrointestinal stromal neoplasm. PMID- 17084731 TI - A new reconstruction of the accessory donor right hepatic artery with interposition of the SMA in liver transplantation. PMID- 17084732 TI - Ovarian mucinous cystadenoma as an etiology of the Zollinger-Ellison Syndrome. PMID- 17084733 TI - Using an internal thoracic artery holder in pancreaticojejunostomy. PMID- 17084734 TI - Decisive presurgical role of MIBI SPECT/CT in identifying within a calcific thyroid nodule the parathyroid responsible for primary hyperparathyroidism. PMID- 17084735 TI - US AIDS coordinator shuns collaboration on neglected disease. PMID- 17084736 TI - Neuroprotection: the end of an era? PMID- 17084737 TI - Dear WHO Executive Board: please keep your nerve. PMID- 17084738 TI - Reviving reproductive health. PMID- 17084739 TI - Putting sexual and reproductive health on the agenda. PMID- 17084740 TI - Cairo after 12 years: successes, setbacks, and challenges. PMID- 17084741 TI - XDR tuberculosis: an indicator of public-health negligence. PMID- 17084742 TI - Antiretroviral programme in rural Uganda. PMID- 17084743 TI - Physicians' assessment and competence: USA and UK. PMID- 17084744 TI - The future for specialists and the Medical Royal Colleges? PMID- 17084745 TI - Mabel Bianco: blazing a trail for women's reproductive rights. PMID- 17084746 TI - Lessons from TGN1412. PMID- 17084747 TI - How clean was the KLEAN trial? PMID- 17084750 TI - ACE inhibitors and aortic rupture. PMID- 17084751 TI - ACE inhibitors and aortic rupture. PMID- 17084754 TI - Premature closure of the Dutch Stent-in I study. PMID- 17084755 TI - Family history in necrotising fasciitis. PMID- 17084756 TI - Intravenous drug use and HIV. PMID- 17084757 TI - Extensively drug-resistant tuberculosis as a cause of death in patients co infected with tuberculosis and HIV in a rural area of South Africa. AB - BACKGROUND: The epidemics of HIV-1 and tuberculosis in South Africa are closely related. High mortality rates in co-infected patients have improved with antiretroviral therapy, but drug-resistant tuberculosis has emerged as a major cause of death. We assessed the prevalence and consequences of multidrug resistant (MDR) and extensively drug-resistant (XDR) tuberculosis in a rural area in KwaZulu Natal, South Africa. METHODS: We undertook enhanced surveillance for drug-resistant tuberculosis with sputum culture and drug susceptibility testing in patients with known or suspected tuberculosis. Genotyping was done for isolates resistant to first-line and second-line drugs. RESULTS: From January, 2005, to March, 2006, sputum was obtained from 1539 patients. We detected MDR tuberculosis in 221 patients, of whom 53 had XDR tuberculosis. Prevalence among 475 patients with culture-confirmed tuberculosis was 39% (185 patients) for MDR and 6% (30) for XDR tuberculosis. Only 55% (26 of 47) of patients with XDR tuberculosis had never been previously treated for tuberculosis; 67% (28 of 42) had a recent hospital admission. All 44 patients with XDR tuberculosis who were tested for HIV were co-infected. 52 of 53 patients with XDR tuberculosis died, with median survival of 16 days from time of diagnosis (IQR 6-37) among the 42 patients with confirmed dates of death. Genotyping of isolates showed that 39 of 46 (85%, 95% CI 74-95) patients with XDR tuberculosis had similar strains. CONCLUSIONS: MDR tuberculosis is more prevalent than previously realised in this setting. XDR tuberculosis has been transmitted to HIV co-infected patients and is associated with high mortality. These observations warrant urgent intervention and threaten the success of treatment programmes for tuberculosis and HIV. PMID- 17084758 TI - Factors that shape young people's sexual behaviour: a systematic review. AB - BACKGROUND: Since nearly half of new HIV infections worldwide occur among young people aged 15-24 years, changing sexual behaviour in this group will be crucial in tackling the pandemic. Qualitative research is starting to reveal how social and cultural forces shape young people's sexual behaviour and can help explain why information campaigns and condom distribution programmes alone are often not enough to change it. We undertook a systematic review to identify key themes emerging from such research, to help inform policymakers developing sexual health programmes, and guide future research. METHODS: We reviewed 268 qualitative studies of young people's sexual behaviour published between 1990 and 2004. We developed a method of comparative thematic analysis in which we coded each document according to themes they contained. We then identified relations between codes, grouping them accordingly into broader overall themes. Documents were classified as either primary or secondary depending on their quality and whether they contained empirical data. From the 5452 reports identified, we selected 246 journal articles and 22 books for analysis. FINDINGS: Seven key themes emerged: young people assess potential sexual partners as "clean" or "unclean"; sexual partners have an important influence on behaviour in general; condoms are stigmatising and associated with lack of trust; gender stereotypes are crucial in determining social expectations and, in turn, behaviour; there are penalties and rewards for sex from society; reputations and social displays of sexual activity or inactivity are important; and social expectations hamper communication about sex. The themes do not seem to be exclusive to any particular country or cultural background, and all themes were present, in varying degrees, in all countries assessed. INTERPRETATION: This study summarises key qualitative findings that help in understanding young people's sexual behaviour and why they might have unsafe sex; policymakers must take these into account when designing HIV programmes. Considerable overlap exists between current studies, which indicates the need to broaden the scope of future work. PMID- 17084759 TI - Adherence to antiretroviral therapy in a home-based AIDS care programme in rural Uganda. AB - BACKGROUND: Poverty and limited health services in rural Africa present barriers to adherence to antiretroviral therapy that necessitate innovative options other than facility-based methods for delivery and monitoring of such therapy. We assessed adherence to antiretroviral therapy in a cohort of HIV-infected people in a home-based AIDS care programme that provides the therapy and other AIDS care, prevention, and support services in rural Uganda. METHODS: HIV-infected individuals with advanced HIV disease or a CD4-cell count of less than 250 cells per muL were eligible for antiretroviral therapy. Adherence interventions included group education, personal adherence plans developed with trained counsellors, a medicine companion, and weekly home delivery of antiretroviral therapy by trained lay field officers. We analysed factors associated with pill count adherence (PCA) of less than 95%, medication possession ratio (MPR) of less than 95%, and HIV viral load of 1000 copies per mL or more at 6 months (second quarter) and 12 months (fourth quarter) of follow-up. FINDINGS: 987 adults who had received no previous antiretroviral therapy (median CD4-cell count 124 cells per muL, median viral load 217,000 copies per mL) were enrolled between July, 2003, and May, 2004. PCA of less than 95% was calculated for 0.7-2.6% of participants in any quarter and MPR of less than 95% for 3.3-11.1%. Viral load was below 1000 copies per mL for 894 (98%) of 913 participants in the second quarter and for 860 (96%) of 894 of participants in the fourth quarter. In separate multivariate models, viral load of at least 1000 copies per mL was associated with both PCA below 95% (second quarter odds ratio 10.6 [95% CI 2.45 45.7]; fourth quarter 14.5 [2.51-83.6]) and MPR less than 95% (second quarter 9.44 [3.40-26.2]; fourth quarter 10.5 [4.22-25.9]). INTERPRETATION: Good adherence and response to antiretroviral therapy can be achieved in a home-based AIDS care programme in a resource-limited rural African setting. Health-care systems must continue to implement, evaluate, and modify interventions to overcome barriers to comprehensive AIDS care programmes, especially the barriers to adherence with antiretroviral therapy. PMID- 17084760 TI - Sexual and reproductive health: a matter of life and death. AB - Despite the call for universal access to reproductive health at the 4th International Conference on Population and Development in Cairo in 1994, sexual and reproductive health was omitted from the Millennium Development Goals and remains neglected (panel 1). Unsafe sex is the second most important risk factor for disability and death in the world's poorest communities and the ninth most important in developed countries. Cheap effective interventions are available to prevent unintended pregnancy, provide safe abortions, help women safely through pregnancy and child birth, and prevent and treat sexually transmitted infections. Yet every year, more than 120 million couples have an unmet need for contraception, 80 million women have unintended pregnancies (45 million of which end in abortion), more than half a million women die from complications associated with pregnancy, childbirth, and the postpartum period, and 340 million people acquire new gonorrhoea, syphilis, chlamydia, or trichomonas infections. Sexual and reproductive ill-health mostly affects women and adolescents. Women are disempowered in much of the developing world and adolescents, arguably, are disempowered everywhere. Sexual and reproductive health services are absent or of poor quality and underused in many countries because discussion of issues such as sexual intercourse and sexuality make people feel uncomfortable. The increasing influence of conservative political, religious, and cultural forces around the world threatens to undermine progress made since 1994, and arguably provides the best example of the detrimental intrusion of politics into public health. PMID- 17084761 TI - Priority setting for health interventions in Mexico's System of Social Protection in Health. AB - Explicit priority setting presents Mexico with the opportunity to match the pressure and complexity of an advancing epidemiological transition with evidence based policies driven by a fundamental concern for how to make the best use of scarce resources to improve population health. The Mexican priority-setting experience describes how standardised analytical approaches to decision making, mainly burden of disease and cost-effectiveness analyses, combine with other criteria--eg, being responsive to the legitimate non-health expectations of patients and ensuring fair financing across households--to design and implement a set of three differentiated health intervention packages. This process is a key element of a wider set of reform components aimed at extending health insurance, especially to the poor. The most relevant policy implications include lessons on the use of available and proven analytical tools to set national health priorities, the usefulness of priority-setting results to guide long-term capacity development, the importance of favouring an institutionalised approach to cost-effectiveness analysis, and the need for local technical capacity strengthening as an essential step to balance health-maximising arguments and other non-health criteria in a transparent and systematic process. PMID- 17084762 TI - Has the management of Alzheimer's disease changed over the past 100 years? PMID- 17084763 TI - Abdominal aortic aneurysm presenting as an epigastric mass. PMID- 17084764 TI - Does academic intervention impact ABS qualifying examination results? AB - OBJECTIVES: To assess the impact of a focused academic support program on American Board of Surgery In-Training Examination (ABSITE) scores and Qualifying Examination (QE) outcomes. METHODS: A mandatory intervention program was begun in April 2001 for residents with ABSITE Total Test (TT) percentiles <31. Program elements included: 1) individual faculty mentoring and personal learning plan 2) QE videotape review sessions 3) Surgical Education and Self-Assessment Program (SESAP) 4) monthly rotation evaluations, and 5) quarterly status feedback. A free medical evaluation was offered. Mock orals participation, educational psychologist consultation, and voluntary followup mentoring were added later. Study data were reviewed for 2003-2005 Chief Residents including ABSITE scores, QE results, conference attendance, rotation Overall Performance ratings, and resident surgeon case volumes. Results were compared for the academic intervention (AI) and no intervention (NI) groups. RESULTS: Fifteen residents graduated during the study period. Eight residents completed nine interventions; seven returned to TT percentiles >30 (7/8, 88%). First post-intervention ABSITE gains were large compared to NI and national peer groups. Standard Score (SS) TT gains were maintained until residency completion by four AI residents. Median AI PGY-5 TT percentile was 32 and three scores were 2 month of age are at risk for postoperative myocardial contractile failure. Myocardial ischemia is associated with a reduction in mitochondrial enzyme activity and have impaired metabolism resulting in decreased postoperative myocardial adenosine triphosphate (ATP) concentrations and increased lactate levels. With this in view, we measured the mitochondrial energy system (respiration and OXPHOS) and to study morphological changes from the right ventricular outflow tract (RVOT) muscle of patients with TOF. METHODS: 30 infants with TOF were studied with age-matched control group consisted of 12 normal patients who died due to extracardiac causes. Mitochondrial respiratory chain complexes, OXPHOS, cytochrome content and ATPase activity were measured by documented standard procedure. Morphological changes examined with a transmission electron microscope. RESULTS: In the presence of glutamate and succinate as substrates, the rate of mitochondrial oxygen consumption was significantly lower in RVOT muscles (p<0.001) by using with and without addition of ADP. The ADP/O ratio indices for glutamate and succinate were not significantly affected. The activities of rotenone-sensitive NADH cytochrome c reductase (complexes I+III), cytochrome c oxidase (complex IV) and the ratio of I and III to II and III complexes (complex I) were significantly lower in TOF (p<0.001). A significant reduction of total cytochrome content and ATPase activity (p<0.001) was noted in study group. Morphological changes were also seen in study group as compared with control. CONCLUSIONS: OXPHOS, mitochondrial respiratory chain complex I, I+III and IV, cytochrome content and ATPase activity are more impaired in RVOT muscles in patients with TOF. PMID- 17084833 TI - Regulation of the Drosophila distal antennal determinant spineless. AB - The transformation of antenna to leg is a classical model for understanding segmental fate decisions in Drosophila. The spineless (ss) gene encodes a bHLH PAS transcription factor that plays a key role in specifying the identity of distal antennal segments. In this report, we identify the antennal disc enhancer of ss and then use enhancer-lacZ reporters to work out how ss antennal expression is regulated. The antennal determinants Distal-less (Dll) and homothorax (hth) are key activators of the antennal enhancer. Dll is required continuously and, when present at elevated levels, can activate the enhancer in regions devoid of hth expression. In contrast, homothorax (hth) is required only transiently both for activation of the enhancer and for specification of the aristal portion of the antenna. The antennal enhancer is repressed by cut, which determines its proximal limit of expression, and by ectopic Antennapedia (Antp). Repression by Antp is not mediated by hth, suggesting that ss may be a direct target of Antp. Finally, we show that ss+ is not a purely passive target of its regulators: ss+ partially represses hth in the third antennal segment and lies upstream of Dll in the development of the maxillary palp primordia. PMID- 17084834 TI - RTK and TGF-beta signaling pathways genes in the sea urchin genome. AB - The Receptor Tyrosine kinase (RTK) and TGF-beta signaling pathways play essential roles during development in many organisms and regulate a plethora of cellular responses. From the genome sequence of Strongylocentrotus purpuratus, we have made an inventory of the genes encoding receptor tyrosine kinases and their ligands, and of the genes encoding cytokines of the TGF-beta superfamily and their downstream components. The sea urchin genome contains at least 20 genes coding for canonical receptor tyrosine kinases. Seventeen of the nineteen vertebrate RTK families are represented in the sea urchin. Fourteen of these RTK among which ALK, CCK4/PTK7, DDR, EGFR, EPH, LMR, MET/RON, MUSK, RET, ROR, ROS, RYK, TIE and TRK are present as single copy genes while pairs of related genes are present for VEGFR, FGFR and INSR. Similarly, nearly all the subfamilies of TGF-beta ligands identified in vertebrates are present in the sea urchin genome including the BMP, ADMP, GDF, Activin, Myostatin, Nodal and Lefty, as well as the TGF-beta sensu stricto that had not been characterized in invertebrates so far. Expression analysis indicates that the early expression of nodal, BMP2/4 and lefty is restricted to the oral ectoderm reflecting their role in providing positional information along the oral-aboral axis of the embryo. The coincidence between the emergence of TGF-beta-related factors such as Nodal and Lefty and the emergence of the deuterostome lineage strongly suggests that the ancestral function of Nodal could have been related to the secondary opening of the mouth which characterizes this clade, a hypothesis supported by functional data in the extant species. The sea urchin genome contains 6 genes encoding TGF-beta receptors and 4 genes encoding prototypical Smad proteins. Furthermore, most of the transcriptional activators and repressors shown to interact with Smads in vertebrates have orthologues in echinoderms. Finally, the sea urchin genome contains an almost complete repertoire of genes encoding extracellular modulators of BMP signaling including Chordin, Noggin, Sclerotin, SFRP, Gremlin, DAN and Twisted gastrulation. Taken together, these findings indicate that the sea urchin complement of genes of the RTK and TGF-beta signaling pathways is qualitatively very similar to the repertoire present in vertebrates, and that these genes are part of the common genetool kit for intercellular signaling of deuterostomes. PMID- 17084835 TI - Transcriptional control of Notch signaling by a HOX and a PBX/EXD protein during vulval development in C. elegans. AB - The Notch signaling pathway controls growth, differentiation and patterning in divergent animal phyla; in humans, defective Notch signaling has been implicated in cancer, stroke and neurodegenerative disorders. Despite its developmental and medical significance, little is known about the factors that render cells to become competent for Notch signaling. Here we show that during vulval development in the nematode Caenorhabditis elegans the HOX protein LIN-39 and its EXD/PBX like cofactor CEH-20 are required for LIN-12/Notch-mediated lateral signaling that specifies the 2 degrees vulval cell fate. Inactivation of either lin-39 or ceh-20 resulted in the misspecification of 2 degrees vulval cells and suppressed the multivulva phenotype of lin-12(n137) gain-of-function mutant animals. Furthermore, both LIN-39 and CEH-20 are required for the expression of basal levels of the genes encoding the LIN-12/Notch receptor and one of its ligands in the vulval precursor cells, LAG-2/Delta/Serrate, rendering them competent for the subsequent lin-12/Notch induction events. Our results suggest that the transcription factors LIN-39 and CEH-20, which function at the bottom of the RTK/Ras and Wnt pathways in vulval induction, serve as major integration sites in coordinating and transmitting signals to the LIN-12/Notch cascade to regulate vulval cell fates. PMID- 17084836 TI - In utero pesticide exposure and childhood morbidity. AB - In humans, immune development begins early in embryonic life and continues throughout the early postnatal period. Although a number of pesticides have been observed to induce developmental immunotoxicity in mice, few human studies have examined the long term effects of in utero pesticide exposure on childhood morbidity. Empirical evidence suggests that the vulnerable period for toxic insults to the developing immune system extends from early gestation to adolescence in humans and animals. Using data from the Ontario Farm Family Health Study, we examined the relationship between farm couple exposures to pesticides during pregnancy and subsequent health outcomes in their offspring, including: persistent cough or bronchitis, asthma, and allergies or hayfever. No strong associations between pesticide exposures during pregnancy and persistent cough or bronchitis, or asthma were found. There was suggestive evidence that allergies and hayfever appeared to be more common in offspring, especially male offspring, exposed to certain specific pesticides during the period of pregnancy. Nevertheless, given the indirect indicators of pesticide exposure used in this study, and the scarcity of human studies on in utero exposure to pesticides and the development of allergies and other child health outcomes, these findings serve primarily to generate hypotheses for future research. PMID- 17084837 TI - The relationships between blood lead levels and serum follicle stimulating hormone and luteinizing hormone in the third National Health and Nutrition Examination Survey. AB - The relationships between blood lead levels and serum follicle stimulating hormone and luteinizing hormone were assessed in a nationally representative sample of women, 35-60 years old, from the third National Health and Nutrition Examination Survey. The blood lead levels of the women ranged from 0.7 to 31.1 microg/dl. The estimated geometric mean was 2.2 microg/dl, and the estimated arithmetic mean was 2.8 microg/dl. As the blood lead level increased across women, the concentration of serum follicle stimulating hormone increased in post menopausal women, women who had both ovaries removed, and pre-menopausal women. The concentration of follicle stimulating hormone decreased in pre-menopausal women who were taking birth control pills. The concentration of luteinizing hormone increased as blood lead level increased in post-menopausal women and women who had both ovaries removed. The lowest concentrations of blood lead at which a relationship was detected were 1.7 microg/dl for follicle stimulating hormone and 2.8 microg/dl for luteinizing hormone. The increase in follicle stimulating hormone and luteinizing hormone in women with no ovaries indicates that lead may act at a non-ovarian site in the female reproductive system, along with a possible effect on the ovaries. PMID- 17084838 TI - Social sexual inequality and sex difference in cancer incidence. AB - Socioeconomic factors play many roles in influencing health including overall health status, lifestyle and occupational exposures, and access to preventive, diagnostic and treatment services. This paper reviews evidence on the geographical distribution of the sex differences in cancer incidence and life expectancy. The analyses reported are at the regional (Italy), continental (Europe), and world-wide scales. In agreement with other contributions on the social epidemiology of cancer, these results indicate that there is a close link between the health of the populations, and socioeconomic and cultural factors, and support the notion that environment contributes strongly to total cancer incidence. Thus, the emphasis for reducing cancer incidence needs to focus more on reducing environmental contributions. In order to improve the health status of the populations, not only applications of the present etiologic knowledge are necessary (for example, it is estimated that around up to 50% of cancers are nowadays technically preventable), but also further research on environmental topics should be stimulated. Within this perspective the indicators of health differences between genders-which are demonstrated to be very sensitive to socioeconomic and cultural factors--can play a very useful role for monitoring environmental factors, and for health planning by agencies and governments. PMID- 17084839 TI - Lead exposure in an urban community: investigation of risk factors and assessment of the impact of lead abatement measures. AB - INTRODUCTION: A battery recycling plant located in an urbanized area contaminated the environment with lead oxides. The Secretary of Environment of the State of Sao Paulo demanded an evaluation of lead exposure among the population in the vicinity of the plant. OBJECTIVES: To assess the lead exposure of children, to propose control measures and evaluate the impact of these measures. METHODS: Cross-sectional study of all children<13 years old in a radius of 1km from the plant responsible for the contamination. Blood lead levels (BLL) were determined for each child and questionnaires were applied to their parents. Mean BLL were compared before and after control measures were implemented. Logistic regression identified risk factors of lead exposure. RESULTS: Of the 850 investigated children, 311 presented BLL above the action limit established by the World Health Organization. Overall, the median BLL was 7.3 micro g/dL and it varied according to age of children (higher among 1-5 years old) and distance of the residence from the plant. Risk factors identified for BLL>10 micro g/dL were: to live in unpaved areas, parent working in the plant, distance from the plant, to play on the ground, pica, and to drink locally produced milk. After control measures were implemented (closing the plant, soil removal, dust vacuum-cleaning in the households, etc.), a reduction of 46% in BLL was observed considering the 241 re-evaluated children with levels >10 micro g/dL. CONCLUSIONS: This study showed that combined abatement measures were effective in reducing BLL in children living close to a contaminating source. These results informed the decision-making process regarding management of contaminated areas in Brazil. PMID- 17084840 TI - The fibroblast growth factor receptor 4 (FGFR4) Arg388 allele correlates with survival in head and neck squamous cell carcinoma. AB - BACKGROUND: The increased expression of the fibroblast growth factor receptor 4 (FGFR4) has been identified in many human cancers. Recently, a single nucleotide polymorphism changing the sense codon 388 from glycine to arginine was identified in the FGFR4 gene. The FGFR4 Arg(388) allele was found to be associated with a poor prognosis for positive node breast cancer, high-grade soft-tissue sarcoma, colon carcinoma, and head and neck squamous cell carcinoma (HNSCC). METHODS: We decided to verify the impact of the FGFR4 Arg(388) allele on survival as well as its association with histoclinical data in 75 cases of HNSCC. The FGFR4 Arg(388) allele was detected by PCR-RFLP and DNA sequencing. RESULTS: The FGFR4 Arg(388) allele was detected in 42.5% of the tumors (37% heterozygous Gly/Arg and 5.5% homozygous Arg/Arg). The presence of at least one Arg allele was significantly correlated with reduced overall survival after 24 months of follow-up. The cases involving the Arg allele presented an increased mortality risk of 2.2 if compared to the non-carrier cases. CONCLUSION: The FGFR4 Arg(388) allele is associated with a shortened survival. PMID- 17084841 TI - RANK ligand signaling modulates the matrix metalloproteinase-9 gene expression during osteoclast differentiation. AB - Osteoclast differentiation is tightly regulated by receptor activator of NF kappaB ligand (RANKL) signaling. Matrix metalloproteinase-9 (MMP-9), a type IV collagenase is highly expressed in osteoclast cells and plays an important role in degradation of extracellular matrix; however, the molecular mechanisms that regulate MMP-9 gene expression are unknown. In this study, we demonstrate that RANKL signaling induces MMP-9 gene expression in osteoclast precursor cells. We further show that RANKL regulates MMP-9 gene expression through TRAF6 but not TRAF2. Interestingly, blockade of p38 MAPK activity by pharmacological inhibitor, SB203580 increases MMP-9 activity whereas ERK1/2 inhibitor, PD98059 decreases RANKL induced MMP-9 activity in RAW264.7 cells. These data suggest that RANKL differentially regulates MMP-9 expression through p38 and ERK signaling pathways during osteoclast differentiation. Transient expression of MMP-9 gene (+1 to 1174 bp relative to ATG start codon) promoter-luciferase reporter plasmids in RAW264.7 cells and RANKL stimulation showed significant increase (20-fold) of MMP 9 gene promoter activity; however, there is no significant change with respect to +1 bp to -446 bp promoter region and empty vector transfected cells. These results indicated that MMP-9 promoter sequence from -446 bp to -1174 bp relative to start codon is responsive to RANKL stimulation. Sequence analysis of the mouse MMP-9 gene promoter region further identified the presence of binding motif ( 1123 bp to -1153 bp) for the nuclear factor of activated T cells 1 (NFATc1) transcription factor. Inhibition of NFATc1 using siRNA and VIVIT peptide inhibitor significantly decreased RANKL stimulation of MMP-9 activity. We further confirm by oligonucleotide pull-down assay that RANKL stimuli enhanced NFATc1 binding to MMP-9 gene promoter element. In addition, over-expression of constitutively active NFAT in RAW264.7 cells markedly increased (5-fold) MMP-9 gene promoter activity in the absence of RANKL. Taken together, our results suggest that RANKL signals through TRAF6 and that NFATc1 is a downstream effector of RANKL signaling to modulate MMP-9 gene expression during osteoclast differentiation. PMID- 17084842 TI - Plasmodium yoelii: distinct CD4(+)CD25(+) regulatory T cell responses during the early stages of infection in susceptible and resistant mice. AB - The outcome of experimental murine infection with different strains of malaria parasites, ranging from spontaneous cure to death, depends largely on the establishment of effective Th1 responses during the early stages of infection. Here we describe the disparity in CD4(+)CD25(+) regulatory T cell (Treg) responses during the early stages of infection with the highly virulent Plasmodium yoelii 17XL strain in susceptible (BALB/c) and resistant (DBA/2) mice. An increased proportion of Tregs 3-4 days post inoculation, co-occurring with elevated IL-10 levels, is observed in BALB/c but not in DBA/2 mice. These findings suggest that Treg proliferation might be causally associated with the suppression of Th1 responses during early malaria infection, leading to increase parasitemia and mortality in BALB/c mice, possibly in an IL-10-dependent manner. PMID- 17084843 TI - Children in Sweden admitted to intensive care after trauma. AB - OBJECTIVE: The aims of this study were to describe the demographics, injuries, mechanisms and severity of injury, prehospital and hospital care during the first 24h, and patient outcome, in the most severely injured children cared for following trauma at a paediatric intensive care unit in Sweden. METHODS: The medical records of 131 traumatised children (0-16 years of age), admitted to the paediatric intensive care unit in Gothenburg from January 1990 to October 2000, were retrospectively examined. Nine internationally recognised scoring systems were used to calculate severity of injury, in order to predict the chances of patient survival. RESULTS: Paediatric trauma was more common in boys (68%). The mean age at injury was 7.9 years (S.D. 4.7 years). Traffic-related accidents (40%) and falls (34%) were the leading causes of injury. Injuries to the head were the most frequent, forming 24% of all injuries. Severity of injury was recorded as an Injury Severity Score median of 14, Trauma Score Injury Severity Score median of 99% and Paediatric Risk of Mortality Score median of 0.69%. The mortality rate was 3%. CONCLUSION: Trauma with admission to a paediatric intensive care unit is rare in a Swedish paediatric population. When cared for at a centre with the necessary facilities and trained personnel, these children have a good chance of survival. PMID- 17084844 TI - Effects of intranasal versus oral hormone therapy on asymmetric dimethylarginine in healthy postmenopausal women: a randomized study. AB - OBJECTIVE: Oral estrogens reduce asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase, and an independent risk factor for cardiovascular disease. This study was conducted to compare the effect on ADMA between intranasal and oral 17beta-estradiol (E2) combined with norethisterone (acetate) (NET(A)) administration in postmenopausal women. METHODS: In a two center, randomized, double-blind, comparative study 90 healthy postmenopausal women (age 56.6+/-4.7 years) received daily continuous combined intranasal E2/NET 175 microg/275 microg (n=47) or oral E2/NETA 1 mg/0.5 mg (n=43) for one year. At baseline, week 12 and 52, plasma concentrations of ADMA, arginine and symmetric dimethylarginine (SDMA) were measured by high-performance liquid chromatography. RESULTS: Oral E2/NETA reduced ADMA concentrations (-7.4%; 95% confidence interval (CI) -10.4 to -4.4%), while intranasal E2/NET had no effect (-0.8%; 95% CI -3.7 to 2.1%) after 52 weeks. In both groups, arginine was transiently decreased compared to baseline at week 12 (intranasal: -6.1%; 95% CI -9.1 to -3.0%; oral: 6.5%; 95% CI -10.9 to -2.1%). Only oral E2/NETA reduced SDMA concentrations. CONCLUSIONS: Oral administration of E2/NETA reduced ADMA and SDMA concentrations, whereas intranasal administration did not. Both treatments transiently reduced arginine. The decrease in ADMA by oral estrogens could be a key phenomenon in the modulation of nitric oxide synthesis by postmenopausal hormone therapy. PMID- 17084845 TI - Knee biomechanics of moderate OA patients measured during gait at a self-selected and fast walking speed. AB - Osteoarthritis (OA) is a chronic disorder resulting in degenerative changes to the knee joint. Three-dimensional gait analysis provides a unique method of measuring knee dynamics during activities of daily living such as walking. The purpose of this study was to identify biomechanical features characterizing the gait of patients with mild-to-moderate knee OA and to determine if the biomechanical differences become more pronounced as the locomotor system is stressed by walking faster. Principal component analysis was used to compare the gait patterns of a moderate knee OA group (n=41) and a control group (n=43). The subjects walked at their self-selected speed as well as at 150% of that speed. The two subject groups did not differ in knee joint angles, stride length, and stride time or walking speed. Differences in the magnitude and shape of the knee joint moment waveforms were found between the two groups. The OA group had larger adduction moment magnitudes during stance and this higher magnitude was sustained for a longer portion of the gait cycle. The OA group also had a reduced flexion moment and a reduced external rotation moment during early stance. Increasing speed was associated with an increase in the magnitude of all joint moments. The fast walks did not, however, increase or bring out any biomechanical differences between the OA and control groups that did not exist at the self-selected walks. PMID- 17084846 TI - Multidimensional liquid chromatography system with an innovative solvent evaporation interface. AB - An orthogonal two-dimensional liquid chromatographic (2D-LC) system was developed by using a vacuum-evaporation loop-type valve interface. Normal-phase liquid chromatography (NPLC) with a bonded CN phase column was used as the first dimension, and reversed-phase liquid chromatography (RPLC) with a C(18) column was used as the second dimension. All the solvents in the loop of the interface were evaporated at 90 degrees C under vacuum conditions, leaving the analytes on the inner wall of the loop. The mobile phase of the second dimension dissolved the analytes in the loop and injected them onto the secondary column, allowing an on-line solvent exchange of a selected fraction from the first dimension to the second dimension. The chromatographic resolution of analytes on the two dimensions was maintained at their optimal condition. Sample loss due to evaporation in the interface was observed that depended on the boiling point of the compound. Separation of sixteen polycyclic aromatic hydrocarbon mixtures and a traditional Chinese medicine Angelica dahurica was demonstrated. PMID- 17084847 TI - Preparation and characterization of poly(methyltetradecylsiloxane) stationary phases immobilized by gamma radiation onto zirconized silica. AB - The preparation of stationary phases with enhanced chemical stability in alkaline eluents has been the principal objective of many chromatographers. New and improved silica substrates and advanced chemical modification methods are among the possibilities being investigated to reach this objective. The present work has evaluated these two possibilities for new stationary phases. First, the silica surface was modified by reaction with zirconium tetrabutoxide to produce zirconized silica particles having about 21% (w/w) of zirconium. Then poly(methyltetradecylsiloxane) (PMTDS) was immobilized onto this surface using different doses (50-120 kGy) of gamma radiation. These new phases were characterized using elemental analysis and infrared and solid-state (29)Si nuclear magnetic resonance (NMR) spectroscopies. These new stationary phases presented column efficiencies of about 68,000 plates m(-1), symmetric peaks for apolar compounds and retention factors that depend on the irradiation dose and show improved stability in high pH mobile phases. The separation of several pharmaceuticals at pH 11 is presented. PMID- 17084848 TI - Determination of endocrine-disrupting compounds in cereals by pressurized liquid extraction and liquid chromatography-mass spectrometry. Study of background contamination. AB - A sensitive method based on pressurized liquid extraction (PLE) and liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) has been developed for the determination in cereal samples of seven endocrine-disrupting compounds: bisphenol A (BPA), 4-tert-butylbenzoic acid (BBA), 4-nonylphenol (NP), 4-tert-butylphenol (t-BP), 2,4-dichlorophenol (DCP), 2,4,5-trichlorophenol (TCP) and pentachlorophenol (PCP). For the PLE procedure, methanol was selected as the extraction solvent. An experimental design approach was applied to optimize other PLE parameters. The recoveries achieved for the all seven compounds were in the 81-104% range, with relative standard deviations of 4-9%. An additional preconcentration step, based on solid-phase extraction (SPE), after the PLE step proved to be a successful way for obtaining a more sensitive method. The detection limits achieved in corn breakfast cereals were in the 0.003-0.013 microg g(-1) range, except for BPA, with a detection limit of 0.043 microg g(-1), for a sample size of 2.5 g. These values are similar to or even lower than currently legislated limits for pesticides in cereals and cereal-based foodstuffs. We also investigated possible contamination during the experimental process by the target compounds released from purified water, plastics, syringes, peristaltic pump tubes, glassware and other laboratory materials in contact with the samples along the analytical process. PMID- 17084849 TI - Effect of the residual silanol group protection on the liquid chromatographic resolution of racemic primary amino compounds on a chiral stationary phase based on optically active (3,3'-diphenyl-1,1'-binaphthyl)-20-crown-6. AB - A liquid chromatographic chiral stationary phase (CSP) based on (3,3'-diphenyl 1,1'-binaphthyl)-20-crown-6, which has been utilized in the resolution of alpha amino acids, amines and amino alcohols, was treated with excess of n octyltriethoxysilane to prepare a new improved CSP. The residual silanol groups of the original CSP were protected by n-octyl groups in the new CSP. The chiral recognition ability of the new CSP was superior to that of the original CSP in the resolution of alpha-amino acids, amines and amino alcohols. Retention factors (k1) for the resolution of alpha-amino acids were lower on the new CSP than on the original CSP while those for the resolution of amines and amino alcohols were higher on the new CSP than on the original CSP. The improved chiral recognition ability of the new CSP and the retention behaviors of the two enantiomers on the new CSP have been rationalized to stem from the removal of the non enantioselective interactions between the analytes and the residual silanol groups of the original CSP and the improved lipophilicity of the CSP. PMID- 17084850 TI - Quantification of estrogenic mycotoxins at the ng/L level in aqueous environmental samples using deuterated internal standards. AB - Because of their pronounced estrogenicity, resorcyclic acid lactones (RALs) are of concern in aqueous environments even at the low ng/L level. Therefore, we developed an accurate, precise and sensitive HPLC-MS/MS method to detect these mycotoxins in different aqueous environmental samples. The compounds investigated included zearalenone (ZON), alpha- and beta-zearalenol, zearalanone as well as alpha- and beta-zearalanol. The use of isotope labelled internal standards (in this case deuterated RAL-analogues) ensured an accurate quantification of the target analytes, independent of matrix compounds interfering with the analytes during ionisation and analyte losses occurring during sample preparation. Sample enrichment was carried out by solid-phase extraction (SPE) using Supelclean Envi 18 cartridges. Absolute method recoveries for all analytes ranged from 95 to 108%, 70 to 102%, and 76 to 109%, method detection limits from 0.5 to 2.1 ng/L, 0.4 to 1.1 ng/L, and 0.8 to 12.4 ng/L and precision from 3 to 14%, 2 to 13% and 4 to 16% in drainage water, river water and wastewater treatment plant (WWTP) effluent, respectively. The method was applied to verify the emission of RALs from a Fusarium graminearum infested crop field into the drainage system. Zearalenone was present in drainage water in concentrations up to 30 ng/L. So far, none of the other five investigated compounds have been detected. PMID- 17084851 TI - Determination of vanillin and related flavor compounds in cocoa drink by capillary electrophoresis. AB - A simple, rapid, and reliable capillary electrophoresis (CE) method using a photodiode array detector determined four flavor components (vanillin, ethylvanillin, 2-methoxyphenol, and 2-ethoxyphenol) in cocoa drink. Simple and rapid sample preparation required only dilution. Separation used 50 mM phosphate buffer and 2 mM cetyltrimethylammonium hydroxide (CTAH) at pH 10 with 10% acetonitrile. Sorbic acid was the internal standard (I.S.). Vanillin and related compounds were determined in 7 min, with the limits of detection at 1.6 microg/ml with S/N > 3 and a quantitation range of 5-500 microg/ml. Recoveries were investigated in cocoa drink samples with four flavor components. Mean recoveries were 96.3-103.8%. Using this method, the four flavor components were determined from cocoa drink, in which vanillin and ethylvanillin were originally contained as a flavoring and into which Bacillus firmus was added. PMID- 17084852 TI - Stabilization of hydroxyl-group-terminated SERS-marker molecules on microAg particles by silanization. AB - Micrometer-sized Ag (microAg) powders are very efficient substrates for both the infrared and Raman spectroscopic characterization of molecular adsorbates assembled on silver. In particular, the Raman spectrum of organic monolayers on microAg powders is a surface-enhanced Raman scattering (SERS) spectrum. To use microAg powders as a core material for constructing molecular sensing/recognition units operating via SERS, it is first necessary to stabilize the SERS-marker molecules that are directly in contact with the microAg powders. One promising strategy is the fabrication of silica shells onto SERS-marker molecules, and herein we demonstrate its feasibility by choosing 4-mercaptophenol (4-MPH) as a model SERS-marker molecule. Due to the presence of the hydroxyl group of 4-MPH, silica was readily deposited onto microAg particles by the base-catalyzed hydrolysis of tetraethyl orthosilicate, and its subsequent condensation, to form a cagelike structure. The formation of silica shells was confirmed with infrared, Raman, and X-ray photoelectron spectroscopy, coupled with field emission scanning electron microscopy. We were able to tune the thickness of silica shells simply by varying the silanization reaction time. PMID- 17084853 TI - PEG400 novel phase description in water. AB - The behavior of hydroxyl-terminated PEG400 in water was investigated by surface tension measurements and 13C NMR as a function of concentration and temperature. PEG400 exhibited a critical aggregative concentration (cac) that evidenced both its amphiphilic character and its aggregation capacity. Moreover, the chemical shifts of the different carbons of the PEG were followed by NMR versus concentration at various temperatures. We observed a plateau between 20 and 35 degrees C at concentrations above 0.2 mol L(-1) and ascribed it to the aggregation process. A good correlation was found between the NMR spectra in the region of aggregation and the cac region in the phase diagram. Our investigations were also focused on the solid-liquid region of the phase diagram at lower temperatures. These experimental data, together with conclusions available in the literature, led us to propose explanations for the conformation/hydration/aggregation in the PEG400-water solutions phenomena. PMID- 17084854 TI - Corrosion of copper in aerated acidic pickling solutions and its inhibition by 3 amino-1,2,4-triazole-5-thiol. AB - Corrosion of copper in aerated acidic chloride pickling (0.5 M HCl) solutions and its inhibition by 3-amino-1,2,4-triazole-5-thiol (ATT) have been investigated using electrochemical techniques and weight-loss measurements, along with Raman spectroscopy. Electrochemical measurements for copper after varied immersion periods of 0, 24, and 48 h showed that the presence of ATT and the increase of its concentration significantly decrease cathodic, anodic, corrosion (j(Corr)) currents and corrosion rates (K(Corr)), as well as the dissolution currents at 300 mV vs Ag/AgCl, while increasing polarization resistance (Rp), degree of surface coverage (theta) and inhibition efficiency (IE%) to a great extent. Weight-loss measurements after different immersion periods of 6 to 48 h revealed that the dissolution of copper decreased to a minimum and the corresponding IE% increased with increasing ATT concentration. The detection of ATT molecules on the copper surface by Raman spectroscopy indicated that inhibition of copper corrosion is achieved by strong adsorption of ATT molecules onto the copper surface. PMID- 17084855 TI - Inorganic metallodielectric materials fabricated using two single-step methods based on the Tollen's process. AB - Two methods for preparing polycrystalline silver shells on colloidal silica spheres are reported. These do not include the use of organic ligands or metal seeding steps and are based on the Tollen's process for silvering glass. Reaction parameters such as temperature and reactant concentrations are adjusted to slow the reaction kinetics, which we find leads to preferential silver growth on the spheres. The resulting shells are polycrystalline and granular, showing highly uniform sphere coverage. Surface morphologies range from sparsely interconnected grains for shells approximately 20 nm thick, to complete (yet porous) shells of interconnected silver clusters which are up to approximately 140 nm in thickness. The extinction spectra of the core-shell materials are markedly different from those of smooth continuous shells, showing clear evidence that the granular shell geometry influences the plasmon resonance of the composite system. Spheres coated with shells 20-40 nm thick are also suitable for colloidal crystallization. Monolayers of self-assembled spheres with long-range ordering are demonstrated. PMID- 17084856 TI - Facile synthesis of superparamagnetic magnetite nanoparticles in liquid polyols. AB - Magnetite nanoparticles have been successfully synthesized in liquid polyols at elevated temperature. Polyol solvent plays a crucial role in determining the morphology and colloidal stability of the resulting particles. The structure and morphology of the nanoparticles were studied using XRD, TEM, SAED, TGA and FTIR. The magnetic properties of the samples were measured using physical properties measurement system (PPMS) of Quantum Design. The results show that as-prepared magnetite nanoparticles are monodisperse, highly crystalline and superparamagnetic at room temperature. The nanoparticles can be easily dispersed in aqueous media and other polar solvents due to coated by a layer of hydrophilic polyol ligands in situ. This approach provides a facile route to prepare magnetite nanoparticles. PMID- 17084857 TI - Dynamics of unfolded polypeptide chains in crowded environment studied by fluorescence correlation spectroscopy. AB - Proteins have evolved to fold and function within a cellular environment that is characterized by high macromolecular content. The earliest step of protein folding represents intrachain contact formation of amino acid residues within an unfolded polypeptide chain. It has been proposed that macromolecular crowding can have significant effects on rates and equilibria of biomolecular processes. However, the kinetic consequences on intrachain diffusion of polypeptides have not been tested experimentally, yet. Here, we demonstrate that selective fluorescence quenching of the oxazine fluorophore MR121 by the amino acid tryptophan (Trp) in combination with fast fluorescence correlation spectroscopy (FCS) can be used to monitor end-to-end contact formation rates of unfolded polypeptide chains. MR121 and Trp were incorporated at the terminal ends of polypeptides consisting of repetitive units of glycine (G) and serine (S) residues. End-to-end contact formation and dissociation result in "off" and "on" switching of MR121 fluorescence and underlying kinetics can be revealed in FCS experiments with nanosecond time resolution. We revisit previous experimental studies concerning the dependence of end-to-end contact formation rates on polypeptide chain length, showing that kinetics can be described by Gaussian chain theory. We further investigate effects of solvent viscosity and temperature on contact formation rates demonstrating that intrachain diffusion represents a purely diffusive, entropy-controlled process. Finally, we study the influence of macromolecular crowding on polypeptide chain dynamics. The data presented demonstrate that intrachain diffusion is fast in spite of hindered diffusion caused by repulsive interactions with macromolecules. Findings can be explained by effects of excluded volume reducing chain entropy and therefore accelerating the loop search process. Our results suggest that within a cellular environment the early formation of structural elements in unfolded proteins can still proceed quite efficiently in spite of hindered diffusion caused by high macromolecular content. PMID- 17084858 TI - Identification and characterization of single chain anti-cocaine catalytic antibodies. AB - Cocaine is a powerful and addictive stimulant whose abuse remains a prevalent health and societal crisis. Unfortunately, no pharmacological therapies exist and therefore alternative protein-based therapies have been examined. One such approach is immunopharmacotherapy, wherein antibodies are utilized to either bind or hydrolyze cocaine thereby blocking it from exerting its euphoric effect. Towards this end, antibodies capable of binding and hydrolyzing cocaine were identified by phage display from a biased single chain antibody library generated from the spleens of mice previously immunized with a cocaine phosphonate transition state analog hapten. Two classes of antibodies emerged based on sequence homology and mode of action. Alanine scanning mutagenesis and kinetic analysis revealed that residues H97, H99, and L96 are crucial for antibodies 3F5 and 3H9 to accelerate the hydrolysis of cocaine. Antibodies 3F1 through 3F4, which are similar to our previously identified 3A6 class of antibodies, catalyze hydrolysis through transition state stabilization by tyrosine or histidine residues H50 and L94. Mutation of either one or both tyrosine residues to histidine conferred hydrolytic activity on previously inactive antibody 3F4. Mutational analysis of residue H50 of antibody 3F3 resulted in a glutamine mutant with a rate enhancement three times greater than wild-type. A double mutant, containing glutamineH50 and lysineH52, showed a tenfold rate enhancement over wild-type. These results indicate the power of initial selection of catalytic antibodies from a biased antibody library in both rapid generation and screening of mutants for improved catalysis. PMID- 17084859 TI - Role of divalent metal cations in ATP hydrolysis catalyzed by the hepatitis C virus NS3 helicase: magnesium provides a bridge for ATP to fuel unwinding. AB - This study investigates the role of magnesium ions in coupling ATP hydrolysis to the nucleic acid unwinding catalyzed by the NS3 protein encoded by the hepatitis C virus (HCV). Analyses of steady-state ATP hydrolysis rates at various RNA and magnesium concentrations were used to determine values for the 15 dissociation constants describing the formation of a productive enzyme-metal-ATP-RNA complex and the four rate constants describing hydrolysis of ATP by the possible enzyme ATP complexes. These values coupled with direct binding studies, specificity studies and analyses of site-directed mutants reveal only one ATP binding site on HCV helicase centered on the catalytic base Glu291. An adjacent residue, Asp290, binds a magnesium ion that forms a bridge to ATP, reorienting the nucleotide in the active site. RNA stimulates hydrolysis while decreasing the affinity of the enzyme for ATP, magnesium, and MgATP. The binding scheme described here explains the unusual regulation of the enzyme by ATP that has been reported previously. Binding of either free magnesium or free ATP to HCV helicase competes with MgATP, the true fuel for helicase movements, and leads to slower hydrolysis and nucleic acid unwinding. PMID- 17084860 TI - Role of a single amino acid in the evolution of glycans of invertebrates and vertebrates. AB - Structures of glycoconjugate N-glycans and glycolipids of invertebrates show significant differences from those of vertebrates. These differences are due largely to the vertebrate beta1,4-galactosyltransferase-1 (beta4Gal-T1), which is found as a beta1,4-N-acetylgalactosaminyltransferase (beta4GalNAc-T1) in invertebrates. Mutation of Tyr285 to Ile or Leu in human beta4Gal-T1 converts the enzyme into an equally efficient beta4GalNAc-T1. A comparison of all the human beta4Gal-T1 ortholog enzymes shows that this Tyr285 residue in human beta4Gal-T1 is conserved either as Tyr or Phe in all vertebrate enzymes, while in all invertebrate enzymes it is conserved as an Ile or Leu. We find that mutation of the corresponding Ile residue to Tyr in Drosophila beta4GalNAc-T1 converts the enzyme to a beta4Gal-T1 by reducing its N-acetylgalactosaminyltransferase activity by nearly 1000-fold, while enhancing its galactosyltransferase activity by 80-fold. Furthermore, we find that, similar to the vertebrate/mammalian beta4Gal-T1 enzymes, the wild-type Drosophila beta4GalNAc-T1 enzyme binds to a mammary gland-specific protein, alpha-lactalbumin (alpha-LA). Thus, it would seem that, during the evolution of vertebrates from invertebrates over 500 million years ago, beta4Gal-T1 appeared as a result of the single amino acid substitution of Tyr or Phe for Leu or Ile in the invertebrate beta4GalNAc-T1. Subsequently, the pre-existing alpha-LA-binding site was utilized during mammalian evolution to synthesize lactose in the mammary gland during lactation. PMID- 17084861 TI - Low resolution crystal structure of Arenicola erythrocruorin: influence of coiled coils on the architecture of a megadalton respiratory protein. AB - Annelid erythrocruorins are extracellular respiratory complexes assembled from 180 subunits into hexagonal bilayers. Cryo-electron microscopic experiments have identified two different architectural classes. In one, designated type I, the vertices of the two hexagonal layers are partially staggered, with one hexagonal layer rotated by about 16 degrees relative to the other layer, whereas in the other class, termed type II, the vertices are essentially eclipsed. We report here the first crystal structure of a type II erythrocruorin, that from Arenicola marina, at 6.2 A resolution. The structure reveals the presence of long continuous triple-stranded coiled-coil "spokes" projecting towards the molecular center from each one-twelfth unit; interdigitation of these spokes provides the only contacts between the two hexagonal layers of the complex. This arrangement contrasts with that of a type I erythrocruorin from Lumbricus terrestris in which the spokes are broken into two triple-stranded coiled coils with a disjointed connection. The disjointed connection allows formation of a more compact structure in the type I architecture, with the two hexagonal layers closer together and additional extensive contacts between the layers. Comparison of sequences of the coiled-coil regions of various linker subunits shows that the linker subunits from type II erythrocruorins possess continuous heptad repeats, whereas a sequence gap places these repeats out of register in the type I linker subunits, consistent with a disjointed coiled-coil arrangement. PMID- 17084863 TI - A mathematical model of the treatment and survival of patients with high-grade brain tumours. AB - More years of life per patient are lost as the result of primary brain tumours than any other form of cancer. The most aggressive of these is known as glioblastoma (GBM). The median survival time of patients with GBM is under 10 months and the outlook has hardly improved over the past 20 years. Generally, these tumours are remarkably resistant to radiotherapy and yet about 2-3% of all GBMs appear to be cured. The objectives of this study were to formulate a mathematical and phenomenological model of tumour growth in a population of patients with GBM to predict survival, and to use the model to extract biological information from clinical data. The model describes the growth of the tumour and the resulting damage to the normal brain using simple concepts borrowed from chemical reaction engineering. Death is assumed to result when the amount of surviving normal brain falls to a critical level. Radiotherapy is assumed to destroy tumour but not healthy brain. Simple rules are included to represent approximately the clinician's decisions about what type of treatment to offer each patient. A population of patients is constructed by assuming that key parameters can be sampled from statistical distributions. Following Monte Carlo simulation, the model can be fitted to data from clinical trials. The model reproduces clinical data extremely accurately. This suggests that the long-term survivors are not a separate sub-population but are the 'lucky tail' of a unimodal distribution. The estimated values of radiation sensitivity (represented as SF2, the survival fraction after 2Gy) suggest the presence of severe hypoxia, which renders cells less sensitive to radiation. The model can predict the probable age distribution of tumours at presentation. The model shows the complicated effects of waiting times for treatment on the survival outcomes, and is used to predict the effects of escalation of radiotherapy dose. The model may aid the design of clinical trials using radiotherapy for patients with GBM, especially in helping to estimate the size of trial required. It is also designed in a generic form, and might be applicable to other tumour types. PMID- 17084862 TI - Selective photoaffinity labeling identifies the signal peptide binding domain on SecA. AB - SecA, an ATPase crucial to the Sec-dependent translocation machinery in Escherichia coli, recognizes and directly binds the N-terminal signal peptide of an exported preprotein. This interaction plays a central role in the targeting and transport of preproteins via the SecYEG channel. Here we identify the signal peptide binding groove (SPBG) on SecA addressing a key issue regarding the SecA preprotein interaction. We employ a synthetic signal peptide containing the photoreactive benzoylphenylalanine to efficiently and specifically label SecA containing a unique Factor Xa site. Comparison of the photolabeled fragment from the subsequent proteolysis of several SecAs, which vary only in the location of the Factor Xa site, reveals one 53 residue segment in common with the entire series. The covalently modified SecA segment produced is the same in aqueous solution and in lipid vesicles. This spans amino acid residues 269 to 322 of the E. coli protein, which is distinct from a previously proposed signal peptide binding site, and contributes to a hydrophobic peptide binding groove evident in molecular models of SecA. PMID- 17084864 TI - Auto-modulation of neuroactive steroids on GABA A receptors: a novel pharmacological effect. AB - GABA(A) receptor function is modulated by various important drugs including neuroactive steroids that act on allosteric modulatory sites and can directly activate GABA(A) receptor channels at high concentrations. We used whole cell patch-clamp recordings and rapid applications of the neuroactive steroid alphaxalone to investigate repetitive steroid effects. Alphaxalone potentiation of submaximal GABA-evoked currents was enhanced significantly by repetitive coapplications at all investigated recombinant isoforms (alpha1beta3delta, alpha1beta3gamma2L, alpha6beta3delta, alpha6beta3gamma2L) and at GABA(A) receptors of differentiated human NT2 neurons. A similar increase of current amplitudes was induced by repetitive applications of a high steroid concentration without GABA. We refer to these reversible effects as auto-modulation because repeated interactions of steroids enhanced their own pharmacological impact at the receptor sites in a time and concentration dependent manner without affecting GABA controls. Pronounced auto-modulatory actions were also measured using the neurosteroid 5alpha-THDOC in contrast to indiplon, THIP, and pentobarbital indicating a steroid specificity. Protein kinase A inhibition significantly reduced alphaxalone auto-modulation at alpha1beta3gamma2L, alpha6beta3gamma2L, and alpha6beta3delta subtypes while it enhanced potentiation at alpha1beta3delta isoforms suggesting a crucial influence of receptor subunit composition and phosphorylation for steroid actions. Especially at extrasynaptic GABA(A) receptor sites containing the delta subunit steroid auto-modulation may have a critical role in enhancing potentiation of GABA-induced currents. PMID- 17084865 TI - Effects of the NMDA-receptor antagonist ketamine on perceptual correlates of long term potentiation within the nociceptive system. AB - We recently reported perceptual correlates of long-term potentiation (LTP) of synaptic strength within the nociceptive system demonstrating the functional relevance of LTP for human pain sensation. LTP is generally classified as NMDA receptor dependent or independent. Here we show that low doses of the NMDA receptor antagonist ketamine (0.25 mg/kg) prevented the long-term increase in perceived pain to electrical test stimuli, which was induced by high-frequency electrical stimulation (HFS) of nociceptive afferents. Whereas in a control experiment HFS led to a stable increase in perceived pain by 51% for the entire observation period of 1h HFS given 4 min after i.v. ketamine was ineffective. In contrast, HFS induced a two-fold increase of pinprick-evoked pain surrounding the HFS site (secondary neurogenic hyperalgesia) in both experiments. Pain evoked by light tactile stimuli (allodynia) was also unaffected by ketamine. These data support the concept that homotopic hyperalgesia to electrical stimulation of the conditioned pathway is a perceptual correlate of NMDA-receptor sensitive homosynaptic LTP in the nociceptive system, e.g. in the spinal cord. Although secondary neurogenic hyperalgesia and allodynia are induced by the same HFS protocol, they involve additional NMDA-receptor insensitive mechanisms of heterosynaptic facilitation. PMID- 17084866 TI - 5-HT3 receptor-mediated photic-like responses of the circadian clock in the rat. AB - Serotonin (5-HT) and 5-HT agonists have various resetting effects on the master clock, located in the suprachiasmatic nucleus (SCN), depending on the species. In rats, they induce photic-like effects on both locomotor activity rhythms and gene expression in the SCN. The 5-HT receptor(s) mediating these effects at circadian time 22 are localized in the SCN, most likely at a presynaptic level, on the retinohypothalamic terminals (RHT) known to convey photic information by releasing glutamate. Indeed, RHT degeneration blocks photic-like effects of a non specific 5-HT agonist, quipazine. However, the 5-HT receptor subtype(s) involved is still unknown, although 5-HT(3) receptor activation is known to induce glutamate release. We thus analyzed the effects of selective 5-HT(3) agonist and antagonist, as well as a specific NMDA receptor antagonist, on different parameters of the clock. This study shows that the 5-HT(3) receptor mediates the resetting effects of quipazine on locomotor activity rhythms. The 5-HT(3) receptor is only partially implicated in quipazine-induced expression of c-FOS, while NMDA receptor inhibition blocks quipazine photic-like effects on both parameters. Taken together, photic-like responses produced by 5-HT stimulation in rats are likely mediated by (presynaptic?) 5-HT(3) receptor activation followed by NMDA receptor activation. PMID- 17084867 TI - Effects of intracerebroventricular infusions of arginine vasopressin in sheep. AB - In sheep, neither the in vivo effect of vasopressin administered by a method other than systemic infusion nor the central effects on behavior from the perspective of stress regulation has been fully elucidated in an intact animal. We examined changes in behavioral, adrenocorticotropic, and autonomic nervous functions after intracerebroventricular infusions of arginine vasopressin (AVP) to elucidate its central role. Intracerebroventricular infusions of AVP (0, 0.12, 1.2 and 12 microg/500 microl/30 min) evoked a dose-related increase in plasma cortisol concentration. There were significant treatment-related effects on the total duration of sham-chewing (Friedman's test, X2=12.75, p=.0052), on the total duration of bar-biting (Friedman's test, X2=15.0, p=.0018), and on the total duration of rubbing (Friedman's test, X2=12.0, p=.0074). AVP 12 microg treatment induced a greater degree of sham-chewing and bar-biting than the other three treatments did (Nemenyi multiple comparisons: p<0.1). These findings indicate, together with our previous findings, that AVP has the same corticotropic potential as corticotropin-releasing hormone infused intracerebroventricularly in equal molar concentrations. Although the degree to which central stress signaling pathways are involved in these responses remains speculative, the relationships between stereotypies and central AVP are of particular interest. PMID- 17084868 TI - Photic and non-photic effects on the daily activity pattern of Mongolian gerbils. AB - The paper analyses the daily activity pattern of Mongolian gerbils with and without access to a running wheel. To evaluate the synchronizing and the masking effects of light, experiments were performed under different photoperiods (L:D=14:10 h and 10:14 h), and light and dark pulses were applied at different phases of the day-night cycle. In order to get a more direct estimate of the central pacemaker of the circadian system, the body temperature rhythm was investigated via implanted transmitters. Without access to a running wheel, the daily activity pattern was bimodal. One peak occurred in the first half of the light time, the other one around the light-dark transition. Also, the gerbils were more active during the light phase as compared to the dark phase. After unlocking the running wheel, the gerbils were active mainly during the dark time. The activity peak in the first half of the light phase remained, the second one shifted by a phase delay into the dark time. These results were found under both LD-regimens. Light during the night nearly completely suppressed running wheel activity, switching off the light during the day time induced wheel running. Whereas wheel running was clearly affected by light and dark pulses, the general activity was not. The body temperature rhythm also shows two peaks, with the second one being bigger and coinciding with the endogenous component of the circadian body temperature rhythm. It was found around light-off. After unlocking the running wheel, the maximum of the body temperature rhythm shifted to the night. This was not primarily a consequence of the changed activity pattern as shown by means of purification analysis. Removing the direct effects of motor activity led to a body temperature curve that could be described by a cosine function, and the delay shift was found also for the purified data, a better estimate of the endogenous circadian component. The wheel-associated increase in nocturnality is not only due to masking effects of wheel-running activity on the body temperature and activity rhythms. It also involves clock-related processes. Changes in the phase preference may serve as an adaptation mechanism to the changes in the animal's natural environment. PMID- 17084869 TI - Apocarotenoid biosynthesis in arbuscular mycorrhizal roots: contributions from methylerythritol phosphate pathway isogenes and tools for its manipulation. AB - During colonization by arbuscular mycorrhizal (AM) fungi plant roots frequently accumulate two types of apocarotenoids (carotenoid cleavage products). Both compounds, C(14) mycorradicin and C(13) cyclohexenone derivatives, are predicted to originate from a common C(40) carotenoid precursor. Mycorradicin is the chromophore of the "yellow pigment" responsible for the long-known yellow discoloration of colonized roots. The biosynthesis of apocarotenoids has been investigated with a focus on the two first steps of the methylerythritol phosphate (MEP) pathway catalyzed by 1-deoxy-D-xylulose 5-phosphate synthase (DXS) and 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR). In Medicago truncatula and other plants the DXS2 isogene appears to be specifically involved in the AM-mediated accumulation of apocarotenoids, whereas in the case of DXR a single gene contributes to both housekeeping and mycorrhizal (apo)carotenoid biosynthesis. Immunolocalization of DXR in mycorrhizal maize roots indicated an arbuscule-associated protein deposition, which occurs late in arbuscule development and accompanies arbuscule degeneration and breakdown. The DXS2 isogene is being developed as a tool to knock-down apocarotenoid biosynthesis in mycorrhizal roots by an RNAi strategy. Preliminary results from this approach provide starting points to suggest a new kind of function for apocarotenoids in mycorrhizal roots. PMID- 17084870 TI - Identification and functional expression of a type 2 acyl-CoA:diacylglycerol acyltransferase (DGAT2) in developing castor bean seeds which has high homology to the major triglyceride biosynthetic enzyme of fungi and animals. AB - Seed oil from castor bean (Ricinus communis) contains high amounts of hydroxy fatty acid rich triacylglycerols (TAGs) that can serve as raw material for production of bio-based products such as nylon, cosmetics, lubricants, foams, and surfactants. Diacylglycerol acyltransferase (DGAT) catalyses the terminal reaction in the acyl-CoA dependent Kennedy pathway of triglyceride biosynthesis. There is still some debate whether there are three or four enzymes in yeast that have DGAT activity and catalyse the synthesis of TAG but of these the DGAT2 homologue Dga1 contributes in a major way to TAG biosynthesis. Here we report on the cloning of a cDNA for DGAT2 from castor bean and prove its biological activity following expression in yeast and enzymatic assays using diricinolein as the acceptor and ricinoleoyl-CoA as the donor. Previous reports of DGAT in castor have focussed on DGAT1 which has little amino acid sequence homology to DGAT2. Expressional studies demonstrate that DGAT2 is 18-fold more highly expressed in seeds than in leaves and shows temporal specific expression during seed development. In contrast, DGAT1 shows little difference in expression in seeds versus leaves. We conclude that in castor bean DGAT2 is more likely to play a major role in seed TAG biosynthesis than DGAT1. PMID- 17084871 TI - Criteria to distinguish between natural situations and illegal use of boldenone, boldenone esters and boldione in cattle 1. Metabolite profiles of boldenone, boldenone esters and boldione in cattle urine. AB - Boldenone is an androgenic steroid that improves the growth and food conversion in food producing animals. In most countries worldwide, this anabolic steroid is forbidden for meat production. Until recently, the control of its illegal use was based either on 17beta-boldenone or 17alpha-boldenone (its main metabolite in cattle) identification in edible tissues, hair, faeces or urine. Recent observations and data tend to demonstrate the natural occurrence (but not ubiquitous) in cattle of these steroids, making the analytical strategy of the control more complicated. We investigated the metabolism of boldenone in cattle after intramuscular and oral treatment of boldenone, boldenone esters and boldione. The central objective was to elucidate the structures of the main metabolites (phase I and phase II) in urine, with main objective to be further in position to compare boldenone urinary profiles of treated and non-treated animals. Nine metabolites have been identified, only four were present whatever the treatment and the administered boldenone source. Nevertheless, all of them have been detected at least once in non-treated animals which did not permit us to use them as biomarkers of an illegal treatment. At last, but not at least, all metabolites were found mainly glucuro-conjugated, and rarely sulfo-conjugated, with the only exception of 17beta-boldenone. Current investigations are showing the absence of 17beta-boldenone sulfoconjugate in non-treated animals; that would permit to distinguish non-treated from treated animals with boldione, boldenone and boldenone esters. PMID- 17084872 TI - Spatially extended host-parasite interactions: the role of recovery and immunity. AB - Techniques for determining the long-term dynamics of host-parasite systems are well established for mixed populations. The field of spatial modelling in ecology is more recent but a number of key advances have been made. In this paper, we use state-of-the-art approximation techniques, supported by simulations, in order to investigate the role of recovery and immunity in spatially structured populations. Our approach is to use correlation models, namely pair-wise models, to capture the spatial relationships of contacts and interactions between individuals. We use the pair-wise framework to address a number of key ecological questions; including, the persistence of endemic limit cycles and regions of parasite-driven extinction--features which differentiate spatial from non-spatial models--and the effects on invasion fitness. We demonstrate a loss of limit cycle behaviour, in addition to an increase in the critical transmissibility and extinction thresholds, when recovery is included. This approach allows for a better analytical understanding of the dynamics of host-parasite interactions and demonstrates the importance of recovery and immunity in local interactions. PMID- 17084873 TI - In vitro screening of 200 pesticides for agonistic activity via mouse peroxisome proliferator-activated receptor (PPAR)alpha and PPARgamma and quantitative analysis of in vivo induction pathway. AB - Peroxisome proliferator-activated receptors (PPARs) are ligand-dependent transcription factors and key regulators of lipid metabolism and cell differentiation. However, there have been few studies reporting on a variety of environmental chemicals, which may interact with these receptors. In the present study, we characterized mouse PPARalpha and PPARgamma agonistic activities of 200 pesticides (29 organochlorines, 11 diphenyl ethers, 56 organophosphorus pesticides, 12 pyrethroids, 22 carbamates, 11 acid amides, 7 triazines, 8 ureas and 44 others) by in vitro reporter gene assays using CV-1 monkey kidney cells. Three of the 200 pesticides, diclofop-methyl, pyrethrins and imazalil, which have different chemical structures, showed PPARalpha-mediated transcriptional activities in a dose-dependent manner. On the other hand, none of the 200 pesticides showed PPARgamma agonistic activity at concentrations 90% phylogenetic sequence conservation), that IRP binding and signal responses vary quantitatively. The structural specificity of each IRE-RNA provides an opportunity for finding small molecule regulators in vitro, and possibly in vivo. The potential of manipulating mRNA function with small molecules targeted to specific RNA regulatory structures, e.g., ferritin mRNA in iron overload, or viral mRNA control structures for replication, is high. PMID- 17084902 TI - Fluconazole susceptibility testing of Candida species by flow cytometry. AB - OBJECTIVE: Currently, antifungal drug susceptibility testing is labor-intensive, limited by delays in obtaining results and high costs. The purpose of this study was to determine the usefulness of flow cytometry (FCM) antifungal drug susceptibility testing as a routine laboratory procedure. METHODS: A total of 24 clinical isolates of Candida spp. and reference strains were tested for susceptibility to fluconazole by FCM using propidium iodide (PI) as an indicator of viability. The minimum inhibitory concentration (MIC) was defined as the lowest concentration of fluconazole that resulted in an increase of 30% in mean channel fluorescence (MCF), compared to the growth control. FCM results were compared with MIC results as determined by the Clinical and Laboratory Standards Institute (CLSI) method. RESULTS: An 8h incubation was sufficient for determination of the MICs. The results by FCM at 8h and the NCCLS methods at 24h showed 87.5% agreement to within two drug dilutions. However, the FCM method is labor-intensive in proportion to the larger number of samples. For Candida lusitaniae, MICs by the FCM method showed poor correlation with the CLSI method. CONCLUSIONS: Further evaluation is necessary to assess the usefulness of FCM as a technique for routine antifungal MIC testing in the clinical laboratory. PMID- 17084904 TI - Aortic depressor nerve function examined in diabetic rats by means of two different approaches. AB - The present study examined in anesthetized rats, 5 or 120 days after the onset of streptozotocin-induced diabetes, the aortic depressor nerve (ADN) function by means of pressure-nerve activity curve (fitted by sigmoidal regression) and cross spectral analysis between mean arterial pressure (MAP) and ADN activity. From the sigmoidal regression curve it was calculated the upper and lower ADN activity plateau, range, average gain and MAP halfway between the lower and upper plateau (MAP50). By means of spectral analysis it was calculated the transfer function magnitude (ratio of ADN activity/MAP) as an index of ADN sensitivity (gain) during induced (withdrawal and reinfusion of blood) slow (0.35 Hz) oscillations of MAP simulating Mayer's waves and spontaneous oscillations (approximately 1.5 Hz) caused by respiratory movement. Diabetic rats exhibited, at 5 or 120 days, lower MAP and heart rate. The parameters calculated by means of the sigmoidal regression curve, as well as the ADN activity gain during slow or spontaneous oscillations of MAP, were similar in diabetic and control rats. In conclusion, it was demonstrated that ADN activity was not altered after 5 or 120 days of experimental diabetes, even though the literature documents, at this time frame of diabetes, a conspicuous derangement of the baroreflex. PMID- 17084903 TI - Manganese neurotoxicity: a focus on the neonate. AB - Manganese (Mn) is an essential trace metal found in all tissues, and it is required for normal amino acid, lipid, protein, and carbohydrate metabolism. While Mn deficiency is extremely rare in humans, toxicity due to overexposure of Mn is more prevalent. The brain appears to be especially vulnerable. Mn neurotoxicity is most commonly associated with occupational exposure to aerosols or dusts that contain extremely high levels (>1-5 mg Mn/m(3)) of Mn, consumption of contaminated well water, or parenteral nutrition therapy in patients with liver disease or immature hepatic functioning such as the neonate. This review will focus primarily on the neurotoxicity of Mn in the neonate. We will discuss putative transporters of the metal in the neonatal brain and then focus on the implications of high Mn exposure to the neonate focusing on typical exposure modes (e.g., dietary and parenteral). Although Mn exposure via parenteral nutrition is uncommon in adults, in premature infants, it is more prevalent, so this mode of exposure becomes salient in this population. We will briefly review some of the mechanisms of Mn neurotoxicity and conclude with a discussion of ripe areas for research in this underreported area of neurotoxicity. PMID- 17084905 TI - Life stress and the course of early-onset bipolar disorder. AB - BACKGROUND: Studies of adult bipolar patients and adolescents with major depression indicate that life stress and mood symptoms are temporally and causally related to one another. This study examined whether levels of life stress predict levels of mood symptoms among bipolar adolescents participating in a treatment development study of family-focused psychoeducation and pharmacotherapy. METHODS: Bipolar adolescents (n=38) who reported a period of acute mood symptoms within the prior 3 months were recruited for a 1-year study of life stress. Clinician-administered evaluations were completed with adolescents and parents at 3-month intervals for up to 12 months, using the UCLA Life Stress Interview and the K-SADS Mania and Depression Rating Scales. RESULTS: Chronic stress in family, romantic and peer relationships was associated with less improvement in mood symptoms over the study year. The frequency of severe, independent life events also predicted less improvement in mood symptoms. Higher levels of chronic stress in family and romantic relationships, and higher severity of independent events, were more strongly associated with mood symptoms among older adolescents. Results were independent of adolescents' psychosocial treatment regimens. LIMITATIONS: The majority of adolescents received family focused psychoeducational treatment and all were being treated with psychotropic medication. The influence of life stress on mood symptoms may have been attenuated by intensive intervention. CONCLUSIONS: Stress is linked to changes in mood symptoms among bipolar adolescents, although correlations between life events and symptoms vary with age. Chronic stress in family, romantic, and peer relationships are important targets for psychosocial intervention. PMID- 17084906 TI - Patterns of alcohol use between genders: a cross-cultural evaluation. AB - INTRODUCTION: Alcohol use by men and women is very much influenced by social habits and customs. Cultural peculiarities and biological differences between the sexes require more focused and standardized studies. The objective was to systematize information on patterns of alcohol use between the sexes. METHOD: A literary review (1972-2004) identified 96 publications (Lilacs, Scielo, Medline) and some related books. RESULTS AND CONCLUSIONS: Men drank more and presented more problems (legal, family, social, clinical, traumas and mortality) associated with alcohol use; the consequences of alcohol use in developing countries with low death rates is even higher. Women can face more discrimination by using alcohol as well as worse health problems when they abuse drinking (liver, pancreas, and central and peripheral nervous system problems, psychiatric comorbidity, etc.); sexual abuse is more commonly associated with women than discussing the different responses to treatment. As for social roles/responsibilities exercised by women, there are indications that marriage, employment, and children have a good influence, discouraging alcohol use, while divorce, unemployment, and no children contribute to higher consumption. For both sexes, religion was a protective factor for alcohol use; acculturation was a strong influence in the pattern of alcohol use, and alcohol worsened the evolution of existing psychiatric disorders. PMID- 17084907 TI - Validation of the Thai Edinburgh Postnatal Depression Scale for screening postpartum depression. AB - This study aimed to validate and determine an appropriate cut-off score on the Thai Edinburgh Postnatal Depression Scale (EPDS) as a screen for postpartum depression. A prospective cohort of postpartum women at 6-8 weeks were tested using the EPDS and clinically interviewed by psychiatrists to establish a DSM-IV diagnosis of major or minor depressions in a university hospital in Southern Thailand. Of 351 postpartum women interviewed, 38 postpartum women met the criteria for depressive disorders, major depression in four women (1%) and minor depressive disorder in 34 women (10%). The area under the curve was 0.84 (95% confidence interval 0.76-0.91). Using an EPDS cut-off sum score of 6/7, major and/or minor depression was detected with a sensitivity of 74%, specificity of 74%, positive predictive value of 26% and negative predictive value of 95%. When the cut-off score was higher, the sensitivity was lower but the specificity was higher. The Thai version of the EPDS is a valid self-report instrument and is useful in Thailand where no other screening instrument for postpartum depression is available. PMID- 17084908 TI - The role of BTB domain-containing zinc finger proteins in T cell development and function. AB - Cell fate specifications during T lymphocyte differentiation result from the orchestrated expression of developmentally regulated genes. Furthermore, epigenetic processes that result in a heritable chromatin structure are required for the maintenance of gene expression programs within cells. More and more is known about the basic mechanisms of T cell development and their diversification into various peripheral T cell subsets. Recent research has begun to provide insight into the interactive network of transcription factors as critical regulators of T lymphocyte differentiation. In the past years several members of the BTB domain-containing family of zinc finger proteins (BTB-ZF) have been described to be important for the development and function of hematopoietic cells, and also to contribute to malignant hematopoiesis. This review will provide a brief overview about the role of BTB-ZF proteins during thymocyte development and T cell function. PMID- 17084909 TI - Glatiramer acetate-specific human CD8(+) T cells: increased IL-4 production in multiple sclerosis is reduced by glatiramer acetate treatment. AB - Glatiramer acetate (GA) is an approved drug for therapy of relapsing remitting MS that acts as a T cell antigen. Here, we report the cloning of HLA restricted, GA specific human CD8(+) T cells. In addition, we analyzed the cytokine profile of GA-reactive CD8(+) T cell lines. Unexpectedly, IL-4 was increased in untreated MS patients as compared to healthy individuals (p<0.001). In GA-treated patients, however, IL-4 (p<0.001), IL-10 (p<0.001) and TNF-alpha (p<0.001) were decreased. Thus, while GA is known to induce a TH2 bias in CD4(+) T cells, we detected a distinct pattern in GA-reactive CD8(+) T cells. PMID- 17084910 TI - Increase of CD8+ T-effector memory cells in peripheral blood of patients with relapsing-remitting multiple sclerosis compared to healthy controls. AB - CCR7 and CD45RA expression on CD4+ and CD8+ T-cells in blood (PB) of 16 patients with multiple sclerosis (MS) and 16 healthy controls and cerebrospinal fluid (CSF) of 10 patients suffering from MS were analysed by flow cytometric measurements. T-cells were divided by their distinct homing potentials and effector-functions in three groups: naive T-cells (CCR7+, CD45RA+), central memory T-cells (TCM) (CCR7+, CD45RA-) and effector memory T-cells (TEM) (CCR7-, CD45RA-). There was a significant increase of CD8+ TEM-cells in PB of MS patients compared to healthy controls, indicating systemic immune activation. Further we found a relative depletion of CD8+ TEM-cells in CSF of MS patients compared to matching blood samples, suggesting that these cells represent the effector arm of the immune response and infiltrate the brain tissue at the sites of inflammation. PMID- 17084911 TI - Role of ERK in cocaine addiction. AB - Cocaine addiction is characterized by compulsive drug-taking behavior and high rates of relapse. According to recent theories, this addiction is due to drug induced adaptations in the cellular mechanisms that underlie normal learning and memory. Such mechanisms involve signaling by extracellular signal-regulated kinase (ERK). As we review here, evidence from rodent studies also implicates ERK in cocaine psychomotor sensitization, cocaine reward, consolidation and reconsolidation of memories for cocaine cues, and time-dependent increases in cocaine seeking after withdrawal (incubation of cocaine craving). The role of ERK in these behaviors involves long-term stable alterations in synaptic plasticity that result from repeated cocaine exposure, and also rapidly induced alterations in synaptic transmission events that acutely control cocaine-seeking behaviors. Pharmacological manipulations that decrease the extent to which cocaine and cocaine cues induce ERK activity might therefore be considered as potential treatments for cocaine addiction. PMID- 17084912 TI - Social attachment in juvenile monkeys with neonatal lesion of the hippocampus, amygdala and orbital frontal cortex. AB - Non-human primates, like humans, develop and maintain social relationships and attachments throughout their life. The first and most crucial relationship in a primate life is that with its mother. Yet, in absence of their biological mother, infant primates form attachment to surrogate mothers. Although, this early attachment is critical for the development of normal species-typical social and emotional skills, the neural substrates underlying the formation of social relationships in primates are still unclear. The present study assessed, in infant rhesus monkeys (Macaca mulatta) reared by human caregivers and social interactions with peers, the effects of bilateral neonatal (1-2 weeks of age) ibotenic acid lesions of the amygdala and hippocampus (N=6 in each group), aspiration lesions of the orbital frontal cortex (N=6) or sham lesions (N=5) on the development of a social attachment with the principal human caregiver. A specific preference for the later was assessed at 11 months of age, in a two choice discrimination task, opposing the principal human caregiver to another familiar human, in a familiar environment. None of the lesions impaired the expression of preferential responses toward the principal human caregiver. Nevertheless, lesions of the orbital frontal cortex led to a weaker preference, suggesting that this structure may play a role in the quality and/or strength of the infant/mother relationships. The present non-human primate findings are discussed in terms of their relevance for autism. PMID- 17084913 TI - Contribution of cutaneous inputs from the hindpaw to the control of locomotion in rats. AB - The influence of reduced feedback from the cutaneous receptors in the hindpaw in rat locomotion is still unclear. To evaluate this question, we conducted a detailed hindlimb kinematic analysis in animals, which suffered complete loss of thermal sensation. Two-dimensional hindlimb kinematics, temporal and spatial measurements, and walking track analysis were performed in rats before and during hypothermic anesthesia. The walking velocity, duration of the step cycle and stance phase, and stride length between the two testing conditions were statistically indistinguishable. Swing phase duration was significantly decreased during sensory loss. Analysis of angular motion revealed an increased hip and knee extension and an ankle joint with increased flexion during the step cycle under plantar anesthesia. Also after plantar cooling, the hip and knee angular velocity was significantly affected along the step cycle. The remarkably geometric similarity of the angle-angle plots obtained in our experiments reflected an interjoint coordination; however, the interpretation of the cyclogram perimeter revealed a larger excursion by the ankle and hip in their respective joint spaces in rats deprived of sensation. Examination of the horizontal position of the ankle with respect to the hip and the extension before toe-off revealed no major changes, whereas, there was a slight decrease in distance of the hip to the ground during sensory loss. Also, the walking tracks revealed a significant functional deficit following reduced cutaneous information of the plantar aspect of the hindpaw. We therefore conclude that sensory feedback from the hindpaw is important in the maintenance of normal rat locomotion. PMID- 17084914 TI - Variations of maternal care alter offspring levels of behavioural defensiveness in adulthood: evidence for a threshold model. AB - Natural variations of maternal care in the rat influence the development of neuronal systems that regulate defensive responses to threat. Thus, as adults, rats that received higher levels of maternal licking/grooming (LG) in infancy display dramatic reductions in burying in the shock-probe test, relative to offspring of low LG mothers. We sought to replicate that finding and determine whether maternal care similarly influences offspring responses to social threat, using the resident-intruder test. We also examined whether maternal LG influences offspring behaviour along a continuum by comparing defensive responses of offspring of mid LG mothers to those of offspring of high LG and low LG mothers. A final goal was to assess whether the reductions in adult offspring reactivity to threat that typically follow corticosterone (CORT) administration to dams across lactation are mediated through CORT-induced changes in maternal care. Adult offspring of high LG mothers spent less time burying the shock-probe, relative to offspring of mid and low LG mothers, whereas offspring of CORT treated mothers did not differ from any group. Similarly, offspring of high LG (but not CORT-treated) mothers displayed fewer defensive responses in the resident-intruder test. Thus, only natural variations of maternal care were associated with individual differences in offspring reactivity to threat. Furthermore, because offspring of mid and low LG mothers displayed equivalent levels of defensive responding in both tests, it appears that a critical threshold of maternal LG is necessary to alter the developmental trajectory of neural systems mediating defensive behaviours. PMID- 17084915 TI - Influence of biotransformation on trophic transfer of the PAH, fluoranthene. AB - The persistence of polycyclic aromatic hydrocarbons (PAHs) in marine sediments may be influenced by benthic invertebrate bioturbation. Through processes such as deposit-feeding and enhancement of microbial metabolic activity PAHs may be remobilized from the sediment compartment, and either transferred to organisms at higher trophic levels or to the overlying water column, both processes inevitably changing the bioavailability of the PAH. Accumulation of contaminants from one level in the food chain to the next depends on feeding rate and assimilation efficiency, two factors that basically vary with food quality and contaminant type. Though it is generally believed that pre-consumptive biotransformation will reduce bioavailability due to the more polar nature of the metabolites compared to the unchanged parent compound, theoretically the decrease in lipophilicity will increase the sediment/food desorption rate in the intestine, and some metabolites will still be lipophilic enough to be absorbed by passive diffusion. We examined the trophic transfer of the PAH, fluoranthene from two closely related polychaete species (i.e., Capitella sp. I and Capitella sp. S), differing in their biotransformation ability, to the predatory polychaete, Nereis virens. We found that N. virens fed the biotransforming species, Capitella sp. I, accumulated significantly more Flu equivalents compared to worms fed Capitella sp. S, which have a very limited biotransformation ability. The dose-specific increase in N. virens intestinal Flu concentration was approximately twice as high in worms fed Capitella sp. I (equation: gut content=7.3 x dose-3.9) compared to worms fed Capitella sp. S (equation: gut content=3.2 x dose+0.6). In addition, we measured DNA damage, using the comet assay, in N. virens intestinal cells after feeding with the two prey species. We did not detect DNA damage above 'background' levels for worms fed either of the two Capitella species, possibly due to relatively low intestinal Flu concentrations in N. virens. Our results indicate that accumulation of PAHs by infaunal organisms may play an important role in the transfer of this type of contaminant to higher trophic levels. Moreover, we observed differences in transfer potential between parent compounds and their respective metabolites, which may influence the fate of these compounds in marine ecosystems. However, from the present study it cannot be concluded whether differences in biotransformation ability among prey species can lead to different effects in their predators. PMID- 17084916 TI - Both the basal transcriptional activity of the GADD45A gene and its enhancement after ionizing irradiation are mediated by AP-1 element. AB - The growth arrest and DNA damage-inducible gene 45A (GADD45A) is involved in the DNA repair, maintenance of genomic stability, cell cycle control and apoptosis, and thus plays an important role in cellular response to DNA damage. The GADD45A gene is responsive to a variety of DNA-damaging agents, including ionizing radiation (IR), methyl methanesulfonate (MMS), and ultraviolet (UV) radiation. It is generally thought that induction of the GADD45A gene after IR exposure is principally p53-dependent, requiring binding of the p53 protein to the p53 recognition sequence in the third intron. However, the involvement of factors other than p53 in transcriptional regulation of the GADD45A gene after IR exposure has not been elucidated. In the present study, we show that the 5' flanking region containing two OCT sites and a CCAAT box, as well as p53 and AP-1 sites in the third intron, are required for the basal transcriptional activity of the reporter gene. In addition, AP-1 recognition element was shown to be involved in the transcriptional enhancement of the GADD45A gene after X-ray irradiation. Electrophoretic mobility shift analysis (EMSA) and Chromatin immunoprecipitation (ChIP) assay revealed that JunD binds to the third intron of the GADD45A gene. These observations suggest that AP-1 complexes containing JunD, in addition to p53, play an important role not only in transcriptional enhancement by IR but also in basal expression of the GADD45A gene via binding to the AP-1 site in the third intron. PMID- 17084918 TI - Calcium signals mediated by STIM and Orai proteins--a new paradigm in inter organelle communication. AB - In all cells Ca2+ signals are key to controlling a spectrum of cellular responses. Ca2+ signals activated by phospholipase C-coupled receptors have two components-rapid Ca2+ release from ER stores followed by slower Ca2+ entry from outside the cell. The coupling process between ER and PM to mediate this "store operated" Ca2+ entry process has remained a molecular and mechanistic mystery. Through a combination of high throughput screening and molecular physiological approaches, the machinery and mechanism of this process have been elucidated. Two proteins are key to the coupling process. STIM1, a single spanning membrane protein with an unpaired Ca2+ binding EF-hand functions as the sensor of ER luminal Ca2+ and through redistribution in the ER transduces information directly to the PM. Orai1, a tetra-spanning PM protein, functions as the highly Ca2+ selective channel in the PM that is gated through interactions with the store activated ER Ca2+ sensor. This molecular pas-de-deux between ER and PM components represents not only a crucial signaling pathway, but also a new paradigm in inter organelle communication. PMID- 17084917 TI - CFTR surface expression and chloride currents are decreased by inhibitors of N WASP and actin polymerization. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) undergoes rapid turnover at the plasma membrane in various cell types. The ubiquitously expressed N-WASP promotes actin polymerization and regulates endocytic trafficking of other proteins in response to signaling molecules such as Rho-GTPases. In the present study we investigated the effects of wiskostatin, an N-WASP inhibitor, on the surface expression and activity of CFTR. We demonstrate, using surface biotinylation methods, that the steady-state surface CFTR pool in stably transfected BHK cells was dramatically decreased following wiskostatin treatment with a corresponding increase in the amount of intracellular CFTR. Similar effects were observed for latrunculin B, a specific actin-disrupting reagent. Both reagents strongly inhibited macroscopic CFTR-mediated Cl(-) currents in two cell types including HT29-Cl19A colonic epithelial cells. As previously reported, CFTR internalization from the cell surface was strongly inhibited by a cyclic-AMP cocktail. This effect of cyclic-AMP was only partially blunted in the presence of wiskostatin, which raises the possibility that these two factors modulate different steps in CFTR traffic. In kinetic studies wiskostatin appeared to accelerate the initial rate of CFTR endocytosis as well as inhibit its recycling back to the cell surface over longer time periods. Our studies implicate a role for N-WASP-mediated actin polymerization in regulating CFTR surface expression and channel activity. PMID- 17084919 TI - 'Full-house' rheumatic heart disease. AB - We report the case and cardiac imaging of a 27 year old African female with every clinical feature of advanced rheumatic heart disease. The case illustrates the continuing life-threatening impact of rheumatic heart disease in young adults. Appropriate diagnosis and management of rheumatic heart disease in the UK remains relevant with increased immigration of at-risk populations. PMID- 17084920 TI - Positive cardiac troponin I and T and chest pain in a patient with iatrogenic hypothyroidism and no coronary artery disease. AB - We report about a 41-year old male patient who presented to the emergency room with acute chest pain, exertion dyspnoea, muscle stiffness, myalgia and adynamia. There was no history of coronary artery disease but known arterial hypertension and insulin dependent diabetes mellitus. Four weeks before submission the patient had been thyroidectomized after he had been diagnosed with papillary thyroid carcinoma and was now awaiting further radioiodine therapy. The thyroid stimulating hormone level was markedly elevated to 67 mU/l (normal range 0.27 4.20 mU/l) and fT4 significantly reduced to 0.19 ng/ml (normal range 0.9-1.9 ng/ml). CK was elevated to 328 U/l, cardiac Troponin I (Stratus CS) above the threshold with 0.13 microg/l and Elecsys third generation troponin T above the threshold with 0.04 microg/l. The electrocardiogram showed a normal sinus rhythm and did not reveal any signs of ST-elevation or -depression. During follow-up a cardiac MRI was performed, showing normal dimensions and function but a very small area of diffuse myocardial damage, atypical of ischemic injury. In coronary angiography normal coronary arteries were found. We conclude that cardiac troponins I and T may be elevated in severe hypothyroidism without coronary artery disease due to diffuse myocardial injury which can be imaged by MRI. PMID- 17084921 TI - Increased aortic stiffness in glycogenosis type 2 (Pompe's disease). AB - BACKGROUND: Pompe's disease, also known as acid maltase deficiency or glycogen storage disease type II, is an autosomal recessive disorder in which deficient activity of the enzyme acid alpha-glucosidase causes intra-lysosomal accumulation of glycogen in muscle and other tissues. The current study was designed to assess aortic stiffness index (beta), as a characteristic of aortic elasticity during transthoracic echocardiography in patients with Pompe's disease. METHODS: A total of 17 patients (age 44+/-8 years, 5 males) with Pompe's disease were studied. Their results were compared to 17 age- and gender-matched controls. In all patients, the ascending aorta was recorded with M-mode echocardiography. Beta was calculated as ln(SBP/DBP)/[(SD-DD)/DD], where SBP and DBP are the systolic and diastolic blood pressures, SD and DD are the systolic and diastolic aortic diameters, and 'ln' is the natural logarithm. RESULTS: Diastolic aortic diameter was 27.4+/-2.4 mm in Pompe patients and 25.6+/-2.7 mm in controls (P<0.05). Systolic aortic diameters did not differ between the groups (29.4+/-2.5 mm vs 28.3+/-2.4 mm, P=ns). Aortic stiffness index (beta) was increased in Pompe patients compared to controls (14.6+/-10.1 vs 5.1+/-2.6, P<0.001). CONCLUSIONS: The results of this study indicate that aortic stiffness is increased in patients with Pompe's disease. This may be due to glycogen storage in the vessel wall causing reduced vascular elasticity. PMID- 17084922 TI - Women with left ventricular ejection fraction < or = 20% have better prognosis than men. AB - Women with advanced heart failure have better prognosis than men (men versus women HR 2.5, 95% CI 1.1-5.5, p=0.03). Although interaction ischemic heart disease-gender was not significant, HR was 1.6 (95% CI 0.5-5.6, p=0.43) for patients with coronary artery disease and 3.4 (95% CI 1.1-10.5, p=0.03) for patients without. PMID- 17084923 TI - Anomalous origin of circumflex artery from right sinus of Valsalva: a rare cause of non-ST elevation syndrome. AB - We present a case with anomalous origin of the left circumflex artery from the right coronary artery ostium, which caused a non-ST elevation coronary syndrome. A review of the literature indicates this to be an extremely rare case. PMID- 17084924 TI - Coronary stent fracture and restenosis in the drug-eluting stent era: do we have clues of management? AB - Despite the fact that the drug-eluting stents (DES) have markedly reduced neointimal proliferation, restenosis is still an open subject. Although DES fracture is very rare, it has been considered as one of the reasons for DES restenosis, however, the mechanism and proper management are not clearly elucidated yet. We describe two cases of sirolimus-eluting stent fracture combined with significant restenosis and suggest the possible revascularization strategies for successful management. PMID- 17084925 TI - Utility of 256-slice cone beam tomography for real four-dimensional volumetric analysis without electrocardiogram gated acquisition. AB - PURPOSE: Current ECG-gated multislice CT can reveal any cardiac phase data, including four-dimensional (4D) images, but cannot acquire the whole heart in one scanning, and arrhythmias impair the quality of the images. We used a prototype 256-slice cone beam CT (Athena, Sony Toshiba), with which the whole heart can be acquired in one scanning. MATERIALS AND METHODS: A pulsating device with contrast material (300 mgI/dl) diluted 10x with saline was moved at 5, 40, 60, 70, and 90 to-and-fro movements/min. Non-ECG-gated 256-slice cone beam CT with 256 x 0.5 mm slice thickness was performed during one to-and-fro motion at each rate. After acquisition, each to-and-fro motion was divided into 20 frames, and each volume was measured and volumetric curves were constructed. RESULTS: Even without ECG gated acquisition, the configuration of the pulsating device at any rate continued to the through plane without any gaps, and the 4D movie of the pulsating device could be observed at any rate except 60 movements/min. Volumes of end-diastole (ED) and end-systole (ES), and ejection fraction (EF) at static state were 81.7, 17.5 ml, and 79% respectively. ED volumes were 81.7, 81.7, 70.3, 63.7, 68.3 and 65.9 ml, ES volumes were 17.5, 30.9, 39.8, 62.7, 55.0 and 43.2 ml, and EF was 79, 62, 43, 2, 19 and 34% at 0, 5, 40, 60, 70, and 90 to-and-fro movements/min, respectively. The ratios of ED volume, using the static state as the reference, were 100, 86, 78, 84 and 81%, those of ES volume were 177, 227, 358, 314 and 247%, and those of EF were 78, 54, 3, 24 and 43% at 5, 40, 60, 70, and 90 to-and-fro movements/min, respectively. From the configuration of volumetric curves, only 5/min could be evaluated. At 60 movements/min, the same device images without any motion were observed in 4D images during one to-and-fro motion. This may be because one to-and-fro time and one scanning time were the same (1 s). CONCLUSION: Even without ECG-gated acquisition, this new 256-slice cone beam CT achieved real 4D analysis of the pulsating device. The ED volume and the configuration of volumetric curve could only be evaluated up to 5 movements/min, but because of poor spatial resolution (1 s/rotation), even at 5 movements/min ES volume tended to be overestimated. As a result, EF tended to be underestimated, which may be improved in the next generation (0.5 s/rotation). PMID- 17084926 TI - Pulmonary venous drainage through a highly vascularized left atrial tumor. AB - We present a patient with symptomatic congestive heart failure due to a left atrial sarcoma infiltrating and apparently occluding the left pulmonary veins. The tumor was highly vascularized and enabled attenuated blood drainage from the left upper and lower pulmonary veins despite intra-operative appearance as completely obliterative, thus avoiding persistent left lung pulmonary edema. The tumor was partially removed and the pathologic findings showed advanced angiogenesis. Malignant tumors may occlude large blood vessels and thus may result in the development of a collateral flow. However, we suggest that highly vascularized tumors (macroscopic totally occlusive) may serve as a sponge and enable attenuated blood flow through the tumor and hence may avoid a complete cut off in blood supply or drainage. PMID- 17084927 TI - Home-based cardiac rehabilitation versus hospital-based rehabilitation: a cost effectiveness analysis. AB - BACKGROUND: Home-based cardiac rehabilitation offers an alternative to traditional, hospital-based cardiac rehabilitation. AIM: To compare the cost effectiveness of home-based cardiac rehabilitation and hospital-based cardiac rehabilitation. METHODS: 104 patients with an uncomplicated acute myocardial infarction and without major comorbidity were randomized to receive home-based rehabilitation (n=60) i.e. nurse facilitated, self-help package of 6 weeks' duration (the Heart Manual) or hospital-based rehabilitation for 8-10 weeks (n=44). Complete economic data were available in 80 patients (48 who received home-based rehabilitation and 32 who received hospital-based rehabilitation). Healthcare costs, patient costs, and quality of life (EQ-5D4.13) were assessed over the 9 months of the study. RESULTS: The cost of running the home-based rehabilitation programme was slightly lower than that of the hospital-based programme (mean (95% confidence interval) difference - 30 pounds sterling (- 45 pounds sterling to - 12 pounds sterling) [-44 euro, -67 euro to -18 euro] per patient. The cost difference was largely the result of reduced personnel costs. Over the 9 months of the study, no significant difference was seen between the two groups in overall healthcare costs (78 pounds sterling, - 1102 pounds sterling to 1191 pounds sterling [-115 euro, -1631 euro to -1763 euro] per patient) or quality adjusted life-years (-0.06 (-0.15 to 0.02)). The lack of significant difference between home-based rehabilitation and hospital-based rehabilitation did not alter when different costs and different methods of analysis were used. CONCLUSIONS: The health gain and total healthcare costs of the present hospital-based and home-based cardiac rehabilitation programmes for patients after myocardial infarction appear to be similar. These initial results require affirmation by further economic evaluations of cardiac rehabilitation in different settings. PMID- 17084928 TI - The effects of multi-hospital systems on hospital prices. AB - US hospital prices are rising again after years of limited growth. We analyze trends in hospital prices during a period of significant price growth (1999-2003) to assess whether hospitals that are part of multi-hospital systems were able to increase their prices faster than non-system hospitals. We find hospitals that were members of multi-hospital systems were able to increase their prices substantially more than comparable non-systems hospitals (34% for large systems and 17% for small systems). Further, we find that the systems effect is not confined to hospitals that have other system member hospitals in their local markets. One possible explanation is that hospitals belonging to non-local multi hospital systems have improved their bargaining position vis-a-vis health plans. PMID- 17084929 TI - Use of molecular techniques to elucidate the mechanisms of action of fungal biocontrol agents: a review. AB - Biological control of fungal plant pathogens appears as an attractive and realistic approach, and numerous microorganisms have been identified as biocontrol agents. There have been many efforts to understand the mechanisms of action of fungal biocontrol agents. Microbiological, microscopic, and biochemical techniques applied over many years have shed light on these mechanisms without fully demonstrating them. More recently, the development of molecular techniques has yielded innovative alternative tools for understanding and demonstrating the mechanisms underlying biocontrol properties. To date, more than 70 publications describe the use of molecular techniques for this purpose. They describe work exploiting targeted or non-targeted gene isolation, gene expression profiling, gene inactivation and/or overexpression, the study of regulatory factors. This work has shed considerable light on mechanisms underlying biocontrol properties. It has also fully demonstrated a number of targeted action mechanisms of some biocontrol agents. This review describes the techniques used in such studies, with their potential and limitations. It should provide a guide for researchers wanting to study the molecular basis of the biocontrol in diverse biocontrol agents. PMID- 17084930 TI - Rapid and efficient screening of a Representational Difference Analysis library using reverse Southern hybridisation: identification of genetic differences between Haemophilus parasuis isolates. AB - Representational Difference Analysis (RDA) is an established technique used for isolation of specific genetic differences between or within bacterial species. This method was used to investigate the genetic basis of serovar-specificity and the relationship between serovar and virulence in Haemophilus parasuis. An RDA clone library of 96 isolates was constructed using H. parasuis strains H425(P) (serovar 12) and HS1967 (serovar 4). To screen such a large clone library to determine which clones are strain-specific would typically involved separately labelling each clone for use in Southern hybridisation against genomic DNA from each of the strains. In this study, a novel application of reverse Southern hybridisation was used to screen the RDA library: genomic DNA from each strain was labelled and used to probe the library to identify strain-specific clones. This novel approach represents a significant improvement in methodology that is rapid and efficient. PMID- 17084931 TI - The urokinase plasminogen activator receptor as a gene therapy target for cancer. AB - Urokinase plasminogen activator (uPA) and/or its receptor (uPAR) are essential for metastasis, and overexpression of these molecules is strongly correlated with poor prognosis in a variety of malignant tumours. Impairment of uPA and/or uPAR function, or inhibition of the expression of these components, impedes the metastatic potential of many tumours. Several approaches have been employed to target uPAR with the aim of disrupting its ligand-independent action or interaction with uPA, including the more recent antigene technology. This review discusses the in vivo use of antigene approaches for downregulating uPAR as a potential therapy for cancer. Preclinical studies are advancing towards the translational phase, provided that established orthotopic tumours, which mimic clinical progression and presentation, are treated using clinically acceptable modes of nucleic acid delivery. PMID- 17084932 TI - Cell age, suspending medium and metal ion influence the susceptibility of Escherichia coli O157:H7 to water-soluble maltose chitosan derivative. AB - The effects of cell age, suspending medium and metal ions on the susceptibility of Escherichia coli O157:H7 to the water-soluble maltose chitosan derivative were investigated. In addition, the leakage of glucose, protein (absorbance at 280 nm) and lactate dehydrogenase (LDH) induced by maltose chitosan derivative in saline solution and deionized water, was examined. Cells of E. coli O157:H7 in the mid exponential phase (6 h) were most susceptible to the chitosan derivative followed by cells in the late-exponential phase (12 h) and stationary phase (24 h). In addition, it was found that the susceptibility of the test organism to the maltose chitosan derivative was less in saline solution than in deionized water. The viable population of E. coli O157:H7 in deionized water containing the maltose chitosan derivative (500 ppm), was reduced from ca 7.6 log cfu/ml to a non-detectable level after 10 h of incubation at 37 degrees C compared to a viable population of ca 6.2 log cfu/ml noted in the chitosan derivative containing-saline solution. After cells of E. coli O157:H7 were exposed to the chitosan derivative in deionized water, a marked increase in the levels of glucose, protein and LDH activity was observed in the supernatant of cell suspension compared to cells of test organism exposed to the saline solution containing chitosan derivative. Metal ions were also found to reduce the antibacterial activity of chitosan derivative. Their effectiveness increased at greater concentrations and varied with the kinds of metal ions with Ba(2+) the most effective and Mg(2+) the least effective. PMID- 17084933 TI - The influence of modified atmospheres and their interaction with water activity on the radial growth and fumonisin B(1) production of Fusarium verticillioides and F. proliferatum on corn. Part I: the effect of initial headspace carbon dioxide concentration. AB - The effect of modified atmospheres on the growth and fumonisin B(1) production of Fusarium verticillioides and Fusarium proliferatum on corn is presented in a series of two papers. In this, the first part, the effect of initial headspace (IH) carbon dioxide concentration and its interaction with water activity (a(w)) on growth and fumonisin B(1) production was evaluated. It was observed that at all a(w) values studied, increase in the IH CO(2) concentration generally resulted in a decrease in the colony growth rate (g, mm day(-1)) and maximum colony diameter (D(max), mm) and an increase in the lag phase duration (lambda, day). Although both a(w) and IH CO(2) concentration had significant and synergistic effects on g, a(w) had the largest effect. As little as 10% IH CO(2) completely inhibited the production of fumonisin B(1) by F. verticillioides. F. proliferatum was more resistant and required 40, 30 and 10% IH CO(2) at a(w) 0.984, 0.951 and 0.930, respectively, to completely inhibit fumonisin B(1) production. These results demonstrate that modified atmospheres containing high CO(2) levels could potentially be employed for the protection of corn from fungal spoilage and mycotoxin contamination during the post-harvest period. PMID- 17084934 TI - The influence of modified atmospheres and their interaction with water activity on the radial growth and fumonisin B(1) production of Fusarium verticillioides and F. proliferatum on corn. Part II: The effect of initial headspace oxygen concentration. AB - This paper is the second in a series of two that describe the effect of modified atmospheres on the growth and mycotoxin production of Fusarium verticillioides and Fusarium proliferatum on corn. In this part, the effect of initial headspace (IH) oxygen concentration and its interaction with water activity (a(w)) on growth and fumonisin B(1) production was investigated. In addition, the impact of vacuum packaging and in-cooperation of O(2) scrubbing sachets was also studied. It was observed that at all a(w) values studied, reduction of IH O(2) concentration from 20 to 2% had no significant effect on the colony growth rate (g, mm d(-1)) and lag phase duration (lambda, d). However, g and lambda were positively and negatively correlated to a(w). The IH O(2) concentration was determined to have a a(w) dependent effect on the oxygen consumption rate. Although the maximum colony diameter (D(max), mm) decreased with the reduction of the IH O(2) level, the greatest mycelial density occurred at 10% IH O(2) for both isolates. This observation was accompanied by a trend of a decrease in the value of the IH O(2) level at which the most fumonisin B(1) was produced from 15 to 5% when the a(w) was decreased from 0.976 to 0.930 for F. verticillioides. For F. proliferatum the optimum conditions for fumonisin B(1) production shifted from 20% at a(w) 0.976 to 10% at both 0.951 and 0.930. Vacuum packaging and the in cooperation of O(2) absorbing sachets completely inhibited the growth of both isolates. These results together with those reported in Part I of the study indicate that O(2) should preferably be completely excluded from modified atmospheres that are employed to protect stored corn from fungal growth and mycotoxin production. PMID- 17084935 TI - Studies on Aspergillus section Flavi isolated from maize in northern Italy. AB - In 2003, for the first time in Italy, significant problems arose with colonization and contamination of maize destined for animal feed with Aspergillus section Flavi and aflatoxins (AFs). This resulted in milk and derived products being contaminated with AFM(1) at levels above the legislative limit. There was little knowledge and experience of this problem in Italy. The objectives of this research were thus to study the populations of Aspergillus section Flavi in six northern Italian regions and obtain information on the relative role of the key species, ability to produce sclerotia, production of the main toxic secondary metabolites, aflatoxins and cyclopiazonic acid, and tolerance of key environmental parameters. A total of 70 strains were isolated and they included the toxigenic species A. flavus and A. parasiticus. A. flavus was dominant in the populations studied, representing 93% of the strains. Seventy percent of strains of Aspergillus section Flavi produced AFs, with 50% of strains also producing cyclopiazonic acid. Sixty-two percent of A. flavus strains and 80% of A. parasiticus were able to produce sclerotia at 30 degrees C. Using 5/2 agar, only 1 strain developed S sclerotia and 19 L sclerotia. With regard to ecological studies, growth of Aspergillus section Flavi was optimal at between 25 and 30 degrees C, while AFB(1) production was optimal at 25 degrees C. Regarding water availability (water activity, a(w)), 0.99 a(w) was optimal for both growth and AFs production, while the only aflatoxin produced in the driest condition tested (0.83 a(w)) was AFB(1). This information will be very useful in identifying regions at risk in northern Italy by linking climatic regional information to levels of fungal contamination present and potential for aflatoxin production in maize destined for animal feed. This would be beneficial as part of a prevention strategy for minimising AFs in this product. PMID- 17084936 TI - Striptease on glass: validation of an improved stripping procedure for in situ microarrays. AB - Microarrays have rapidly become an indispensable tool for gene analysis. Microarray experiments can be cost prohibitive, however, largely due to the price of the arrays themselves. Whilst different methods for stripping filter arrays on membranes have been established, only very few protocols are published for thermal and chemical stripping of microarrays on glass. Most of these protocols for stripping microarrays on glass were developed in combination with specific surface chemistry and different coatings for covalently immobilizing presynthesized DNA in a deposition process. We have developed a method for stripping commercial in situ microarrays using a multi-step procedure. We present a method that uses mild chemical degradation complemented by enzymatic treatment. We took advantage of the differences in biochemical properties of covalently linked DNA oligonucleotides on in situ synthesized microarrays and the antisense cRNA hybridization probes. The success of stripping protocols for microarrays on glass was critically dependent on the type of arrays, the nature of sample used for hybridization, as well as hybridization and washing conditions. The protocol employs alkali hydrolysis of the cRNA, several enzymatic degradation steps using RNAses and Proteinase K, combined with appropriate washing steps. Stripped arrays were rehybridized using the same protocols as for new microarrays. The stripping method was validated with microarrays from different suppliers and rehybridization of stripped in situ arrays yielded comparable results to hybridizations done on unused, new arrays with no significant loss in precision or accuracy. We show that stripping of commercial in situ arrays is feasible and that reuse of stripped arrays gave similar results compared to unused ones. This was true even for biological samples that show only slight differences in their expression profiles. Our analyses indicate that the stripping procedure does not significantly influence data quality derived from post-primary hybridizations. The method is robust, easy to perform, inexpensive, and results after reuse are of comparable accuracy to new arrays. PMID- 17084937 TI - Finite element-based probabilistic analysis tool for orthopaedic applications. AB - Orthopaedic implants, as well as other physical systems, contain inherent variability in geometry, material properties, component alignment, and loading conditions. While complex, deterministic finite element (FE) models do not account for the potential impact of variability on performance, probabilistic studies have typically predicted behavior from simplified FE models to achieve practical solution times. The objective of this research was to develop an efficient and versatile probabilistic FE tool to quantify the effect of uncertainty in the design variables on the performance of orthopaedic components under relevant conditions. Key aspects of the computational tool developed include parametric and automated FE model creation for changes in dimensional variables, efficient solution using the advanced mean-value (AMV) reliability method, and identification of the most significant design variables. Two orthopaedic applications are presented to demonstrate the ability of the computational tool to efficiently and accurately represent component performance. PMID- 17084938 TI - Comparison of the effectiveness of trauma services provided by secondary and tertiary hospitals in Malaysia. AB - STUDY OBJECTIVE: The trauma services provided by 6 hospitals operating at 2 levels of care (4 secondary or district general hospitals and 2 tertiary care hospitals) in Malaysia are compared in terms of mortality and disability for direct admissions to emergency departments to test the hypothesis that care at a tertiary care hospital is better than at a district general hospital. METHODS: All cases were recruited prospectively for 1 year. The hospitals were purposefully selected as typical for Malaysia. There are 3 primary outcome measures: death, musculoskeletal impairment, and disability at discharge. Adjustment was made for potential covariates and within-hospital clustering by using multivariable random-effects logistic regression analysis. RESULTS: For direct admissions, logistic-regression-identified odds of dying were associated with older age (>55 years), odds ratio (OR) 1.9 (95% confidence interval [CI] 1.3 to 2.8); head injury, OR 2.7 (95% CI 1.9 to 3.9); arrival by means other than ambulance, OR 0.6 (95% CI 0.4 to 0.8); severe injuries (Injury Severity Score >15) at a district general hospital, OR 45.2 (95% CI 27.0 to 75.7); severe injuries at a tertiary care hospital, OR 11.2 (95% CI 7.3 to 17.3); and admission to a tertiary care hospital compared to a district general hospital if severely injured (Injury Severity Score >15), OR 0.2 (95% CI 0.1 to 0.4). Admission to a tertiary care hospital was associated with increased odds of disability (OR 1.9; 95% CI 1.5 to 2.3) and musculoskeletal impairment (OR 3.5; 95% CI 2.7 to 4.4) at discharge. CONCLUSION: Care at a tertiary care hospital was associated with reduced mortality (by 83% in severe injuries), but with a higher likelihood of disability and impairment, which has implications for improving access to trauma services for the severely injured in Malaysia and other low- and middle-income settings. PMID- 17084939 TI - Acute hemolytic anemia with acanthocytosis associated with high-dose misoprostol for medical abortion. AB - We report a case of acute hemolytic anemia in a 21-year-old Nigerian woman after high-dose misoprostol (4 mg), used for medical abortion. The major causes of inherited or immune hemolytic anemia were excluded. The patient's peripheral blood smear showed acanthocytes and anisopoikilocytosis, which progressively disappeared in the days postingestion. We evaluated RBC features, and we observed reduced RBC Na+ and K+ content and abnormalities in membrane cation transport pathways and in Ca2+ activated K+ channel (Gardos channel), suggesting possible direct effects of misoprostol on RBCs. Although further studies need to be carried out, the present case suggests that high-dose misoprostol, a prostaglandin E1 analogue, severely affects RBC features and causes an acquired acute hemolytic anemia, which is self-limited when misoprostol is withdrawn. PMID- 17084940 TI - Trauma care and limited resources. PMID- 17084941 TI - The impact of B-type natriuretic peptide in addition to troponin I, creatine kinase-MB, and myoglobin on the risk stratification of emergency department chest pain patients with potential acute coronary syndrome. AB - STUDY OBJECTIVE: The emergency department (ED) evaluation of chest pain patients with potential acute coronary syndrome is limited by the initial sensitivity of cell injury markers. BNP is increased during myocardial ischemia and is associated with adverse outcomes. We determine whether the addition of B-type natriuretic peptide (BNP) to troponin I, creatine kinase-MB (CK-MB), and myoglobin increases the sensitivity and negative predictive value (NPV) for acute myocardial infarction, acute coronary syndrome, and 30-day adverse events among chest pain patients with potential acute coronary syndrome. METHODS: A convenience sample of patients aged 30 years or older and presenting to an urban academic ED with nontraumatic chest pain, thus prompting an ECG, was enrolled, and consent was obtained. Blood samples were drawn at 0 and 90 minutes. Thirty day follow-up was performed for all enrolled patients. Main outcomes were acute myocardial infarction, acute coronary syndrome, and 30-day events (death, acute myocardial infarction, or revascularization). BNP cutoffs were derived from receiver operator characteristics curves. The sensitivity, specificity, positive predictive value (PPV), and NPV with 95% confidence intervals (CIs) were calculated with and without BNP. Differences in sensitivity and specificity with the addition of BNP were calculated with 95% CIs, and McNemar's test was performed to compare sensitivities and specificities. RESULTS: Four hundred twenty-six patients were enrolled and analyzed. The cohort was 54.7+/-13.9 years old, 47.7% men, and 63.5% black. The outcomes were acute myocardial infarction, 39 (9.2%), acute coronary syndrome, 101 (23.7%), and 30-day adverse cardiovascular events 52 (12.2%). BNP cutoffs derived were 51, 31, and 31 pg/mL for acute myocardial infarction, acute coronary syndrome, and 30-day events, respectively. The addition of BNP showed increased sensitivity at the cost of decreased specificity for all 3 outcomes, as follows: (1) acute myocardial infarction: sensitivity: 87.2% (95% CI 72.6% to 95.7%) to 97.4% (95% CI 86.5% to 100%), difference 10.3% (95% CI-0.2% to 24.6%), P=.125; specificity: 62.3% (95% CI 57.2% to 67.1%) to 47.8% (95% CI 42.7% to 52.9%), difference 14.5% (95% CI 11.1% to %18.4), P<.0001; (2) acute coronary syndrome: sensitivity: 75.2% (95% CI 65.7% to 83.3%) to 88.1% (95% CI 80.2% to 93.7%), difference 12.9% (95% CI 7.0% to 21.0%), P=.0002; specificity: 68.0% (95% CI 62.6% to 73.0%) to 48.6% (95% CI 43.1% to 54.2%), difference 19.4% (95% CI 15.2% to 24.1%), P<.0001; (3) 30-day events: sensitivity: 71.2% (95% CI 56.9% to 82.9%) to 88.5% (95% CI 76.6% to 95.7%), difference 17.3% (95% CI 7.7% to 30.3%), P=.004; specificity: 61.8% (95% CI 56.6% to 66.7%) to 43.9% (95% CI 38.8% to 49.0%), difference 17.9% (95% CI 14.2% to 22.2%), P<.0001. There were trends toward increased NPV and decreased PPV for all outcomes, and the addition of BNP achieved a NPV of 99.5% (95% CI 97.0% to 100%) compared with 98.0% (95% CI 95.3% to 99.3%) for acute myocardial infarction. CONCLUSION: The addition of BNP as a dichotomous test to troponin I, CK-MB, and myoglobin produces increased sensitivity at a cost of decreased specificity for acute myocardial infarction, acute coronary syndrome, and 30-day adverse events. Because of this tradeoff, BNP cannot be recommended for use among all ED chest pain patients. However, the improved sensitivity may make this test useful in selected cohorts when the decreased specificity is less important. PMID- 17084942 TI - Patterns of ecstasy-associated hyponatremia in California. AB - STUDY OBJECTIVE: We describe the clinical characteristics of patients with ecstasy- (3,4-methylenedioxymethamphetamine [MDMA]) associated hyponatremia (serum sodium level <130 mmol/L) reported to the California Poison Control System during a 5-year period and determine whether a sex difference exists among patients with ecstasy-associated hyponatremia and hyponatremia-associated adverse outcomes. METHODS: We performed a retrospective review of cases involving ecstasy intoxication reported to the California Poison Control System and recorded in its computerized database from January 1, 2000, through October 9, 2005. We excluded cases that did not involve MDMA exposure or in which there were no symptoms or were minimal effects only. Confirmation of exposure to MDMA was based on history of use and, when available, urine toxicology testing results positive for MDMA or amphetamine derivatives. Hyponatremia was defined as a measured serum sodium level less than 130 mmol/L. RESULTS: A total of 1,436 cases potentially involving ecstasy were reported to the California Poison Control System during the 5-year study period, of which 891 were excluded according to the criteria described above. Of the 545 cases that met inclusion criteria, 296 (54.3%) were women and 249 (45.7%) were men. There were 188 cases (34.5%) with a documented serum sodium level, of which 73 (38.8%) reported hyponatremia (Na <130 mmol/L). Of the 73 subjects with hyponatremia, 55 (75.3%) were women and 18 (24.7%) men; of the 115 nonhyponatremic subjects, 50 (43.5%) were women and 65 (56.5%) were men. Among patients with a documented serum sodium level, female sex was associated with increased odds of hyponatremia (odds ratio [OR] 4.0; 95% confidence interval [CI] 2.1 to 7.6). Among women, those with hyponatremia demonstrated increased odds of coma (OR 3.9; 95% CI 1.2 to 12.9), whereas among men, no increased odds of hyponatremia-associated coma were observed (OR 0.8; 95% CI 0.15 to 4.0). CONCLUSION: Female sex was associated with increased odds of hyponatremia and increased odds of hyponatremia-associated coma among persons with ecstasy intoxication and a documented serum sodium level reported to the California Poison Control System from 2000 to 2005. Multiple potential confounders, including spectrum bias, incomplete laboratory data, and individual differences in study subject characteristics, prevent determination of causality about sex differences in the incidence of ecstasy-associated hyponatremia and its complications. PMID- 17084943 TI - Knowledge translation: closing the evidence-to-practice gap. AB - Knowledge translation describes any activity or process that facilitates the transfer of high-quality evidence from research into effective changes in health policy, clinical practice, or products. This increasingly important discipline attempts to conceptually combine elements of research, education, quality improvement, and electronic systems development to create a seamless linkage between interventions that improve patient care and their routine implementation in daily clinical practice. We outline the gap between research and practice and present a case study of an emergency medicine example of validated evidence that has failed to achieve widespread implementation. The authors describe a model of organization of evidence and its relationship with the process that links research from the scientific endeavor to changes in practice that affect patient outcomes. Obstacles to evidence uptake are explored, as well as the limitations of current educational strategies. Innovative strategies in realms such as computerized decision support systems designed to enhance evidence uptake are also described. The potential interface between knowledge translation and continuous quality improvement, as well as the role for bedside tools, is also presented. Research in knowledge translation includes studies that attempt to quantify and understand the discrepancies between what is known and what is done, as well as those that examine the impact and acceptability of interventions designed to narrow or close these gaps. Sentinel examples in this line of research conducted in the emergency department setting are described. PMID- 17084944 TI - Aspiration pneumonitis requiring intubation after procedural sedation and analgesia: a case report. AB - Emergency department (ED) procedural sedation and analgesia is widely and routinely performed; serious complications are rare. We describe the first reported case of aspiration during procedural sedation in the ED. Although our patient required endotracheal intubation and critical care admission, there was no adverse long-term outcome. Given that there were no apparent predisposing factors, we believe it is crucial for emergency physicians to routinely anticipate the possibility of such a complication during each sedation event. PMID- 17084945 TI - Synthesis, radiolabeling and receptor binding of [3H][(1S,2R)ACPC2]endomorphin-2. AB - Previously, we have shown that substitution of Pro(2) for cis-2 aminocyclopentanecarboxylic acid, ACPC in endomorphin-2 results in an analogue with greatly augmented proteolytic stability, high mu-opioid receptor affinity and selectivity. We now report the synthesis and biochemical characterization of [(3)H][(1S,2R)ACPC(2)]endomorphin-2 with a specific activity of 1.41 TBq/mmol (38.17 Ci/mmol). Specific binding of [(3)H][(1S,2R)ACPC(2)]endomorphin-2 was saturable and of high affinity with an equilibrium dissociation constant, K(d)=1.80+/-0.21nM and receptor density, B(max)=345+/-27 fmol x mg protein(-1) at 25 degrees C in rat brain membranes. Similar affinity values were obtained in kinetic and displacement assays. Both Na(+) and Gpp(NH)p decreased the affinity proving the agonist character of the radioligand. [(3)H][(1S,2R)ACPC(2)]endomorphin-2 retained the mu-specificity of the parent peptide. The new radioligand will be a useful tool to map the topographical requirements of mu-opioid peptide binding due to its high affinity, selectivity and enzymatic stability. PMID- 17084946 TI - How do glial-neuronal interactions fit into current neurotransmitter hypotheses of schizophrenia? AB - Evidence is accumulating that the exclusive dopamine hypothesis of schizophrenia has to be abandoned. Instead, a more integrative approach combines different neurotransmitter systems, in which glutamatergic, GABAergic and dopaminergic pathways interact. This paradigm shift coincides with the recognition that, while typical and modern atypical antipsychotic drugs have efficiently controlled the dramatic psychotic symptoms of schizophrenia, their impact on negative and cognitive symptoms is negligible. Indeed, cognitive decline is now believed to represent the core of schizophrenic morbidity and in this context, impairment of glutamate and more specifically NMDA function is of major importance. Given that astrocytes are important in controlling glutamate homeostasis, it is necessary to assign a significant role to glial-neuronal interactions in the pathophysiology of schizophrenia. Indeed, recent data from several animal and human studies corroborate this notion. This review outlines current neurotransmitter hypotheses and evidence for glial impairment in schizophrenia. Furthermore, findings from recent studies of (13)C nuclear magnetic resonance spectroscopy in experimental models of schizophrenia and NMDA hypofunction are presented and their implications for future research on glial-neuronal interactions discussed. PMID- 17084947 TI - Synthesis, in vitro antibacterial and antifungal evaluations of 2-amino-4-(1 naphthyl)-6-arylpyrimidines. AB - A series of 2-amino-4-(1-naphthyl)-6-arylpyrimidines have been synthesized and characterized by IR, NMR, MS, elemental analyses and evaluated for in vitro antibacterial and antifungal activities. Some of the compounds were found to be active against a limited panel of bacteria and fungi. In particular, compounds 4b and 4e were found to be the most effective analogs against the tested bacterial and fungal strains. PMID- 17084948 TI - Integrated scholarship in nursing: An individual responsibility or collective undertaking. PMID- 17084949 TI - Genetic diversity of pneumococcal surface protein A of Streptococcus pneumoniae meningitis in German children. AB - Pneumococcal surface protein A (PspA) is a possible candidate for the development of a pneumococcal vaccine that has the potential to offer a broad range of protection. PspA genes of pneumococcal meningitis isolates (n=40) isolated as part of an ongoing population-based nation-wide surveillance program on invasive pneumococcal disease in children in Germany were analyzed to expand our knowledge on the distribution of PspA families of this important vaccine candidate in Germany. The serotype distribution of the strains was as follows: serotype 4 (n=3), 6B (5), 9V (2), 14 (8), 18C (6), 19F (5), 23F (6), and 7F (5). The pspA genes of these strains could be assigned to 2 families containing 20 pneumococcal strains each. Family I could be subdivided into 2 clades with 17 strains in clade 1 and 3 strains in clade 2, and family II could be subdivided into 3 subgroups (clades 3-5) containing 16, 3, and 1 strain, respectively. Pneumococcal serotypes were distributed evenly over all clades and families. Interestingly, the distribution of the PspA gene families in Germany was seen to differ slightly that found in other countries. PMID- 17084950 TI - Hand transplantation: the state-of-the-art. AB - The feasibility of hand transplantation has been demonstrated, both surgically and immunologically. Levels of immunosuppression comparable to regimens used in solid organ transplantation are proving sufficient to prevent graft loss. Many patients have achieved discriminative sensibility and recovery of intrinsic muscle function. In addition to restoration of function, hand transplantation offers considerable psychological benefits. The recipient's pre-operative psychological status, his motivation and his compliance with the intense rehabilitation programme are key issues. While the induction of graft specific tolerance represents a hope for the future, immunosuppression currently remains necessary and carries significant risks. Hand transplantation should, therefore, only be considered a therapeutic option for a carefully selected group of patients. PMID- 17084951 TI - Deconstructing sickle cell disease: reappraisal of the role of hemolysis in the development of clinical subphenotypes. AB - Hemolysis, long discounted as a critical measure of sickle cell disease severity when compared with sickle vaso-occlusion, may be the proximate cause of some disease complications. New mechanistic information about hemolysis and its effects on nitric oxide (NO) biology and further examination of the subphenotypes of disease requires a reappraisal and deconstruction of the clinical features of sickle cell disease. The biology underlying clinical phenotypes linked to hemolysis may increase our understanding of the pathogenesis of other chronic hemolytic diseases while providing new insights into treating sickle cell disease. The pathophysiological roles of dysregulated NO homeostasis and sickle reticulocyte adherence have linked hemolysis and hemolytic rate to sickle vasculopathy. Nitric oxide binds soluble guanylate cyclase which converts GTP to cGMP, relaxing vascular smooth muscle and causing vasodilatation. When plasma hemoglobin liberated from intravascularly hemolyzed sickle erythrocytes consumes NO, the normal balance of vasoconstriction:vasodilation is skewed toward vasoconstriction. Pulmonary hypertension, priapism, leg ulceration and stroke, all subphenotypes of sickle cell disease, can be linked to the intensity of hemolysis. Hemolysis plays less of a role in the vaso-occlusive-viscosity complications of disease like the acute painful episode, osteonecrosis of bone and the acute chest syndrome. Agents that decrease hemolysis or restore NO bioavailability or responsiveness may have potential to reduce the incidence and severity of the hemolytic subphenotypes of sickle cell disease. Some of these drugs are now being studied in clinical trials. PMID- 17084952 TI - Integrating conventional science and aboriginal perspectives on diabetes using fuzzy cognitive maps. AB - There is concern among Aboriginal communities in Canada that conventional approaches to the treatment of diabetes are ineffective in part because they fail to recognize the local Aboriginal perspective on the causal determinants of diabetes. While this shortcoming has been recognized, there have been no explicit attempts to practically define these perspectives and prescribe how conventional health management might be altered to incorporate them. In part, this may be due to difficulties in communicating Aboriginal perspectives in a manner which permits incorporation into conventional science-based frameworks. Here we use fuzzy cognitive mapping (FCM) as a technique to represent and compare Canadian Aboriginal and conventional science perspectives on the determinants of diabetes as contained in published articles drawn from a search of Medline and PubMed (1966-2005). The FCM allows for a detailed description of the complex system of culture, spirituality and balance at the root of the Aboriginal view. It also highlights how, for these less tangible factors, it is possible to identify more concrete stressors and outcomes which are amenable to management and monitoring. This preliminary comparison of conventional and Aboriginal views also demonstrates the potential for FCM as a technique to extract, compare and integrate perspectives of different knowledge systems into health management and policy development. PMID- 17084953 TI - Family context of mental health risk in Tsunami-exposed adolescents: findings from a pilot study in Sri Lanka. AB - Using survey data from 325 Tsunami-exposed adolescents and mothers from two villages in southern Sri Lanka, this pilot study investigated influences of Tsunami exposure and subsequent psychosocial losses on adolescent depressive and post-traumatic stress disorder (PTSD) symptoms. Findings generally support the study hypotheses: disaster exposure (for example experiences of property destruction and deaths of close others) contributes to depressive and PTSD symptoms in adolescents. Findings also show that psychosocial losses associated with Tsunami exposure, such as prolonged displacement, social losses, family losses, and mental health impairment among mothers, contribute to depressive and PTSD symptoms in adolescents. Results suggest that the influence of Tsunami exposure on adolescent mental health operates partially through Tsunami-related psychosocial losses. As expected, positive mother-child relationships provide a compensatory influence on both depressive and PTSD symptoms of adolescents. In addition, high levels of depressive symptoms among mothers increases the detrimental influence of other Tsunami-related psychosocial losses on adolescent mental health. These preliminary findings suggest ways to improve ongoing recovery and reconstruction programs and assist in formulating new programs for families exposed to both the Tsunami and other natural disasters. More importantly, findings from this pilot study emphasize the urgent need for larger systematic studies focusing on mental health following disaster exposure. PMID- 17084954 TI - Drinking of Salvia officinalis tea increases CCl(4)-induced hepatotoxicity in mice. AB - In a previous study, the drinking of a Salvia officinalis tea (prepared as an infusion) for 14 days improved liver antioxidant status in mice and rats where, among other factors, an enhancement of glutathione-S-transferase (GST) activity was observed. Taking in consideration these effects, in the present study the potential protective effects of sage tea drinking against a situation of hepatotoxicity due to free radical formation, such as that caused by carbon tetrachloride (CCl(4)), were evaluated in mice of both genders. Contrary to what was expected, sage tea drinking significantly increased the CCl(4)-induced liver injury, as seen by increased plasma transaminase levels and histology liver damage. In accordance with the previous study, sage tea drinking enhanced significantly GST activity. Additionally, glutathione peroxidase was also significantly increased by sage tea drinking. Since CCl(4) toxicity results from its bioactivation mainly by cytochrome P450 (CYP) 2E1, the expression level of this protein was measured by Western Blot. An increase in CYP 2E1 protein was observed which may explain, at least in part, the potentiation of CCl(4)-induced hepatotoxicity conferred by sage tea drinking. The CCl(4)-induced hepatotoxicity was higher in females than males. In conclusion, our results indicate that, although sage tea did not have toxic effects of its own, herb-drug interactions are possible and may affect the efficacy and safety of concurrent medical therapy with drugs that are metabolized by phase I enzymes. PMID- 17084955 TI - Lack of toxic effect of technical azadirachtin during postnatal development of rats. AB - Azadirachtin, a biopesticide has been evaluated for its possible toxic effects during postnatal development of rats over two generations. Rats were fed 100, 500 and 1000ppm technical azadirachtin through diet which is equivalent to 5, 25 and 50mg/kg body weight of rats. Technical azadirachtin has not produced any adverse effects on reproductive function and data were comparable to control animals over two generations. There were no toxicological effect in parent rats as evidenced by clinical signs of toxicity, enzymatic parameters like AST, ALT, ALP, S. bilirubin, S. cholesterol, total protein and histopathology of liver, brain, kidney and testes/ovary. The litters of F(1B) and F(2B) generations were devoid of any morphological, visceral and teratological changes. The percent cumulative loss and growth index of pups were also comparable to respective controls in successive growth period of 0, 4, 7, 14 and 21 days in two generations. There were no major malformations in fetuses while some insignificant minor skeletal variations like missing 5th sternebrae and bipartite thoracic centre found were not compound or dose related. No significant pathomorphological changes were observed in liver, kidney, brain and gonads of F(2B) pups. In conclusion rats fed technical azadirachtin showed no evidence of cumulative effects on postnatal development and reproductive performance over two generations. Absence of any major adverse reproductive effects in adults as well as in 21 days old pups of F(2B) generation suggest the safe use of technical azadirachtin as a biopesticide. PMID- 17084956 TI - Protective effects of quercetin against phenylhydrazine-induced vascular dysfunction and oxidative stress in rats. AB - Oxidative stress is a major contributor to the development of vascular dysfunction found in various pathological conditions. Quercetin, one of the potent antioxidant bioflavonoid compounds, has been shown to alleviate oxidative injury by modulation of gene expression leading to suppression of production of reactive oxygen and nitrogen species and conferring an antiapoptotic activity. The aim of the present study was to investigate the protective effects of quercetin in a model of phenylhydrazine (PHZ)-induced oxidant stress, vascular dysfunction and hemodynamic disturbance in rats. Male Sprague-Dawley rats were administered quercetin orally (25 or 50mg/kg/day) for 6 days. On day four, all animals except those in the normal control group, were administered PHZ intraperitoneally. The results showed that PHZ induced severe hemolysis. The mean arterial pressure and hindlimb vascular resistance of PHZ-control rats were markedly decreased compared to normal controls. Treatment with quercetin significantly improved arterial blood pressure and peripheral vascular resistance. Vascular responsiveness to bradykinin, acetylcholine, and phenylephrine in PHZ-control rats was dramatically suppressed and quercetin restored these responses in a dose-dependent manner. Quercetin partially protected blood glutathione, suppressed plasma malondialdehyde levels, and largely suppressed nitric oxide metabolites and superoxide anion production. These results provide the first evidence for the role of the flavonoid, quercetin, in the alleviation of vascular dysfunction in an animal model of PHZ induced oxidant stress. PMID- 17084957 TI - Dose and Hg species determine the T-helper cell activation in murine autoimmunity. AB - Inorganic mercury (mercuric chloride--HgCl(2)) induces in mice an autoimmune syndrome (HgIA) with T cell-dependent polyclonal B cell activation and hypergammaglobulinemia, dose- and H-2-dependent production of autoantibodies targeting the 34 kDa nucleolar protein fibrillarin (AFA), and systemic immune complex deposits. The organic mercury species methylmercury (MeHg) and ethylmercury (EtHg--in the form of thimerosal) induce AFA, while the other manifestations of HgIA seen after treatment with HgCl(2) are present to varying extent. Since these organic Hg species are converted to the autoimmunogen Hg(2+) in the body, their primary autoimmunogen potential is uncertain and the subject of this study. A moderate dose of HgCl(2) (8 mg/L drinking water--internal dose 148 micro gHg/kg body weight [bw]/day) caused the fastest AFA response, while the induction was delayed after higher (25 mg/L) and lower (1.5 and 3 mg/L) doses. The lowest dose of HgCl(2) inducing AFA was 1.5 mg/L drinking water which corresponded to a renal Hg(2+) concentration of 0.53 micro g/g. Using a dose of 8 mg HgCl(2)/L this threshold concentration was reached within 24 h, and a consistent AFA response developed after 8-10 days. The time lag for the immunological part of the reaction leading to a consistent AFA response was therefore 7-9 days. A dose of thimerosal close to the threshold dose for induction of AFA (2 mg/L drinking water--internal dose 118 micro gHg/kg bw per day), caused a renal Hg(2+) concentration of 1.8 micro g/g. The autoimmunogen effect of EtHg might therefore be entirely due to Hg(2+) formed from EtHg in the body. The effect of organic and inorganic Hg species on T-helper type 1 and type 2 cells during induction of AFA was assessed as the presence and titre of AFA of the IgG1 and IgG2a isotype, respectively. EtHg induced a persistent Th1-skewed response irrespectively of the dose and time used. A low daily dose of HgCl(2) (1.5-3 mg/L) caused a Th1-skewed AFA response, while a moderate dose (8 mg/L) after 2 weeks resulted in a balanced or even Th2-skewed response. Higher daily doses of HgCl(2) (25 mg/L) caused a balanced Th2-Th1 response already from onset. In conclusion, while metabolically formed Hg(2+) might be the main AFA-inducing factor also after treatment with EtHg, the quality of the Hg-induced AFA response is modified by the species of Hg as well as the dose. PMID- 17084958 TI - Environmental whole farm management plans: their character, diversity, and use as agri-environmental indicators in New Zealand. AB - This paper reports on a nationwide survey of environmental farm plan programmes and plan characteristics, towards determining New Zealand's (NZ's) capability and eligibility for reporting on the Organisation for Economic Cooperation and Development's (OECD) environmental whole farm management plan agri-environmental indicator. Considerations regarding the validity of the indicator are discussed. Environmental farm planning in NZ is widespread, fragmented, and diverse. Nine of NZ's 16 regional authorities have environmental farm plan programmes involving 20 different types of environmental farm plans. Potentially half of NZ's total area of farmland is included, but this is reduced by limited and selective use of environmental farm plans as policy instruments. Three authorities are capable of readily reporting the number and coverage of their environmental farm plans; six would need to manually compile the information. Compliance monitoring is mostly informal, but necessary for checking the implementation of plan works and actions. Eligibility of NZ environmental farm plans cannot be determined until the OECD define acceptance standards. A contemporary definition of environmental farm plans is proposed, along with additional criteria necessary to ensure such plans are credible indicators of issue ownership, adoption of best management practice, and landscape changes intended to improve environmental quality. PMID- 17084959 TI - Seeking answers on the quest for effective nonsurgical treatment of Peyronie's disease. PMID- 17084960 TI - A placebo-controlled comparison of the efficiency of triple- and monotherapy in category III B chronic pelvic pain syndrome (CPPS). AB - OBJECTIVES: To perform a prospective, placebo-controlled study to examine the efficacy of alpha-blocker compared with triple therapy (alpha-blocker, anti inflammatory, and muscle relaxant) in the treatment of Category IIIB chronic pelvic pain syndrome (Category IIIB CPPS). MATERIALS AND METHODS: The study was conducted between September 2004 and December 2005, and included 90 treatment naive patients, aged 22-42 yr (mean age: 29.1+/-5.2) with Category IIIB CPPS, who were randomized into three groups: group 1, alpha-blocker; group 2, combination of alpha-blocker, anti-inflammatory, and muscle relaxant; group 3, placebo once daily. The patients were treated for 6 mo and were followed up for a further 6 mo. Changes from baseline in the total and domain scores of the NIH Chronic Prostatitis Symptom Index (NIH-CPSI) were evaluated. The primary criterion for response was scoring 50% reduction in NIH-CPSI pain score. RESULTS: The NIH-CPSI initial and sixth-month total scores were 23.1 and 10.7, respectively, in group 1, and 21.9 and 9.2, respectively, in group 2. The initial and sixth month scores remained stable in group 3 (22.9 and 21.9, respectively). There was no statistically significant difference between two treatment arms with respect to efficiency of treatment (p>0.05). The responses in groups 1 and 2 were found durable at the end of 12 mo. CONCLUSIONS: We found that alpha-blocker monotherapy was as effective and safe as triple therapy in the treatment of Category IIIB CPPS. PMID- 17084961 TI - Declarative modeling of a neurulation-like process. AB - MGS is an experimental programming language dedicated to the modeling and the simulation of a special kind of discrete dy- namical systems. Dynamical systems with a dynamical structure (or (DS)(2)) arise when the state space is not fixed a priori but is jointly computed with the current state during the simulation. In this case the evolution function is often given through local rules that drive the interaction between some system components. MGS offers a new kind of data structure, topological collections, to describe the state of a dynamical system, and a new kind of control structure, transformations, to express local and discrete evolution laws. These two notions permit an easy specification of (DS)(2). We propose in this paper a presentation of the MGS language and its main contributions. We show that various topological collections can be unified using concepts developed in combinatorial algebraic topology: cellular complexes and topological chains. Then we apply the notions brought by MGS to model and simulate the first step towards the simulation of the neurulation process in developmental biology where a sheet of cells evolves to a neural tube. It is a direct description of the modification of the topology of an arbitrary structure expressed in terms of local discrete evolution laws. PMID- 17084962 TI - Control of neural chaos by synaptic noise. AB - We study neural automata - or neurobiologically inspired cellular automata - which exhibits chaotic itinerancy among the different stored patterns or memories. This is a consequence of activity-dependent synaptic fluctuations, which continuously destabilize the attractor and induce irregular hopping to other possible attractors. The nature of these irregularities depends on the dynamic details, namely, on the intensity of the synaptic noise and the number of sites of the network, which are synchronously updated at each time step. Varying these factors, different regimes occur, ranging from regular to chaotic dynamics. As a result, and in absence of external agents, the chaotic behavior may turn regular after tuning the noise intensity. It is argued that a similar mechanism might be on the basis of self-controlling chaos in natural systems. PMID- 17084963 TI - Granulosa cell expression of G1/S phase cyclins and cyclin-dependent kinases in PMSG-induced follicle growth. AB - Follicular development involves a complex orchestration of granulosa cell proliferation and differentiation. It is becoming increasingly apparent that the rate of granulosa cell proliferation declines as follicles reach the large antral status, prior to an ovulatory gonadotropin stimulus, although a precise time course and mechanism for this decline has not been described. The goal of the present study was to characterize granulosa cell proliferation following the onset of antral follicle growth in PMSG-primed immature rats, with emphasis on G1/S phase cyclins and cyclin-dependent kinases. Flow cytometric analysis demonstrated that the percentage of granulosa cells in S phase peaked 24-30 h post-PMSG and declined to control levels 48 h after PMSG administration. Expression of both Cyclin D2 and Cdk 4 was highest 12h post-PMSG and decreased to control levels by 48 h. In addition, Cdk 2 protein increased transiently 12-24h after PMSG. Cyclin E expression increased significantly by 12h but remained elevated through 48 h, and multiple isoforms of Cyclin E were observed with increased proliferation. Both Cdk 4 and Cdk 2 activity parallel protein expression, although, changes in Cdk 2 were more marked. Levels of mRNA for the cell cycle inhibitors p21CIP1 and p27KIP1 increased significantly by 48 h post PMSG. These results demonstrate that PMSG-stimulated movement of granulosa cells across the G1/S boundary during follicle growth is transient. In addition, the control of granulosa cell proliferation may reside through the regulation of both Cdk 2 and Cdk 4. PMID- 17084964 TI - Family history of hypertension is not an independent genetic factor predisposing to ischemic stroke subtypes. AB - OBJECTIVES: The effect of family history of hypertension (FHx(HT)), as a genetic factor predisposing to some ischemic stroke subtypes and independently of hypertension as risk factor, has never been studied. METHODS: A positive FHx(HT) was searched in 472 patients (312 men, 160 women; mean [S.D.] age, 67.9 [11.4] years) consecutively admitted for a first-ever ischemic stroke (348) or transient ischemic attack (TIA) (124) due to small vessel disease (SVD, 180), large vessel disease (156), or cardioembolism (136). In this population and in three age bands, the genetically transmitted vascular risk factors, age, gender and the ischemic stroke subtypes were compared between those with and without a positive FHx(HT). RESULTS: None of the risk factors was associated with a positive FHx(HT), except for hypertension in the whole population (68.9% vs. 48.7%; p=0.001) and in the <65 year-old patients (72.5% vs. 39.5%; p<0.001). Regarding the stroke subtypes, a positive FHx(HT) was only associated with SVD in the subgroup of <65 year-old patients (odd ratios (OR), 2.07; 95% confidence interval (CI), 1.01 to 4.25; p=0.045). However, this finding disappeared in a logistic regression analysis, which only retained hypertension as independently associated with SVD. CONCLUSIONS: FHx(HT) is not an independent genetic factor predisposing to some aetiological stroke subtypes. PMID- 17084965 TI - Frequent epigenetic inactivation of chromosome 3p candidate tumor suppressor genes in gallbladder carcinoma. AB - Gallbladder carcinoma (GBC) is a highly malignant neoplasm that represents the leading cause of death for cancer in Chilean females. There is limited information about the molecular abnormalities involved in its pathogenesis. We have identified a number of molecular changes in GBC, including frequent allelic losses at chromosome 3p regions. Four distinct 3p sites (3p12, 3p14.2, 3p21.3 and 3p22-24) with frequent and early allelic losses in the sequential pathogenesis of this neoplasm have been detected. We investigated epigenetic and genetic abnormalities in GBC affecting 6 candidate tumor suppressor genes (TSG) located in chromosome 3p, including DUTT1 (3p12), FHIT (3p14.2), BLU, RASSF1A, SEMA3B and hMLH1 (3p21.3). DNA extracted from frozen tissue obtained from 50 surgical resected GBCs was examined for gene promoter methylation using MSP (methylation specific PCR) technique after bisulfite treatment in all 6 genes. In addition, we performed PCR-based mutation examination using SSCP in FHIT and RASSF1A genes and loss of heterozygosity (LOH) analysis using microdissected tissue in a subset of tumors for the 3p21.3 region with 8 microsatellite markers. A very high frequency of GBC methylation was detected in SEMA3B (46/50, 92%) and FHIT (33/50, 66%), intermediate incidences in BLU (13/50, 26%) and DUTT1 (11/50, 22%) and very low frequencies in RASSF1A (4/50, 8%) and hMLH1 (2/50, 4%). Allelic loss at 3p21.3 was found in nearly half of the GBCs examined. We conclude that epigenetic inactivation by abnormal promoter methylation is a frequent event in chromosome 3p candidate TSGs in GBC pathogenesis, especially affecting genes SEMA3B (3p21.3) and FHIT (3p14.2). PMID- 17084966 TI - Nuclear localization of Survivin renders HeLa tumor cells more sensitive to apoptosis by induction of p53 and Bax. AB - Clinical studies have shown that nuclear expression of the inhibitor of apoptosis protein Survivin in tumor cells predicted a favorable prognosis whereas cytosolic localized protein caused a decreased overall survival. Therefore Survivin's subcellular localization may be important for its anti-apoptotic capacity. To address this question, we investigated localization and function of Survivin in normal human lung fibroblasts (NHLFs) and HeLa tumor cells. NHLFs of early passages expressed Survivin in the nucleus and were highly sensitive to C2 ceramide, which induces the mitochondrial apoptotic pathway. In contrast, NHLFs at higher passages relocated Survivin to the cytosol and became more resistant to C2 ceramide. Blocking nuclear export of Survivin by leptomycin B in HeLa cells increased susceptibility to C2 ceramide. In addition, transduction of HeLa cells with Survivin fused to a nuclear localization signal augmented basal expression levels of p53 and Bax and enhanced sensitivity for intrinsic apoptosis. Those findings suggest that a predominant nuclear localization of Survivin increases the sensitivity for pro-apoptotic stimuli, whereas nuclear export enables Survivin to fulfill its inhibitor of apoptosis function. A therapeutic intervention which holds Survivin in the nucleus of tumor cells might improve cancer therapy. PMID- 17084967 TI - A correlation to estimate the velocity of convective currents in boilover. AB - The mathematical model proposed by Kozanoglu et al. [B. Kozanoglu, F. Ferrero, M. Munoz, J. Arnaldos, J. Casal, Velocity of the convective currents in boilover, Chem. Eng. Sci. 61 (8) (2006) 2550-2556] for simulating heat transfer in hydrocarbon mixtures in the process that leads to boilover requires the initial value of the convective current's velocity through the fuel layer as an adjustable parameter. Here, a correlation for predicting this parameter based on the properties of the fuel (average ebullition temperature) and the initial thickness of the fuel layer is proposed. PMID- 17084968 TI - An empirical model for parameters affecting energy consumption in boron removal from boron-containing wastewaters by electrocoagulation. AB - In this study, it was investigated parameters affecting energy consumption in boron removal from boron containing wastewaters prepared synthetically, via electrocoagulation method. The solution pH, initial boron concentration, dose of supporting electrolyte, current density and temperature of solution were selected as experimental parameters affecting energy consumption. The obtained experimental results showed that boron removal efficiency reached up to 99% under optimum conditions, in which solution pH was 8.0, current density 6.0 mA/cm(2), initial boron concentration 100mg/L and solution temperature 293 K. The current density was an important parameter affecting energy consumption too. High current density applied to electrocoagulation cell increased energy consumption. Increasing solution temperature caused to decrease energy consumption that high temperature decreased potential applied under constant current density. That increasing initial boron concentration and dose of supporting electrolyte caused to increase specific conductivity of solution decreased energy consumption. As a result, it was seen that energy consumption for boron removal via electrocoagulation method could be minimized at optimum conditions. An empirical model was predicted by statistically. Experimentally obtained values were fitted with values predicted from empirical model being as following; [formula in text]. Unfortunately, the conditions obtained for optimum boron removal were not the conditions obtained for minimum energy consumption. It was determined that support electrolyte must be used for increase boron removal and decrease electrical energy consumption. PMID- 17084969 TI - Central nitric oxide inhibition modifies metabolic adjustments induced by exercise in rats. AB - The influence of the central nervous system on metabolic function is of interest in situations deviating from basal states, such as during exercise. Our previous study in rats demonstrated that central nitric oxide (NO) blockade increases metabolic rate, reducing mechanical efficiency during exercise. To assess the role of brain nitric oxide in the plasma glucose, lactate and free fatty acids (FFAs) concentrations of rats submitted to an incremental exercise protocol on a treadmill until fatigue, 1.43 micromol (2 microl) of N(omega)-nitro-l-arginine methyl ester (L-NAME, n=6), a NO synthase inhibitor, or 2 microl of 0.15M NaCl (SAL, n=6) was injected into the lateral cerebral ventricle (icv) of male Wistar rats immediately before exercise (starting at 10 m/min, with increments of 1m/min every 3 min until fatigue, 10% inclination). Blood samples were collected through a chronic jugular catheter at rest and during exercise until fatigue. During exercise, the L-NAME-treated animals had the following metabolic response compared to controls: (1) an increased hyperglycemic response during the first 60% of time to fatigue; (2) higher plasma lactate levels; and (3) a significant transitory increase in plasma free fatty acids during the dynamic phase of exercise that returned to basal levels earlier than controls during the steady state phase of exercise. In addition L-NAME-treated rats fatigued earlier than controls. The data indicate that the inhibition of the brain nitrergic system induced by icv L-NAME treatment disrupted the accuracy of the neural mechanism that regulates plasma glucose and free fatty acids mobilization during exercise in rats. PMID- 17084970 TI - The anticonvulsant effects of allopregnanolone against amygdala-kindled seizures in female rats. AB - It has long been known that the steroid hormone progesterone has anticonvulsant actions. These have been documented both in animals and humans. In 2003, we reported that progesterone's first metabolite, 5alpha-dihydroprogesterone (5alpha DHP), has strong anticonvulsant effects in amygdala-kindled female rats. These occur without sedation, and involve suppression of the kindled amygdala focus, as well as the secondarily generalized kindled seizure. The purpose of this study was to investigate the anticonvulsant actions of progesterone's secondary metabolite, allopregnanolone, in the amygdala kindling model. Adult female Wistar rats were implanted with chronic indwelling electrodes in the right amygdala, and kindled to 30 stage 5 seizures. Varying doses of allopregnanolone were then administered to each subject in randomized order, and the effects on the kindled amygdala focus and the secondarily generalized kindled seizure were observed. Immediately before each drug trial, ataxia was rated using the Loscher scale. Complete suppression of the generalized kindled convulsion was seen in all subjects, with an ED(50) of 1.1 mg/kg. Ataxia--scored as Loscher stage 2 or higher--was seen at higher doses, with a TD(50) of 8.6 mg/kg. The therapeutic index for suppression of the generalized convulsion was 7.8. Even at the highest doses tested, however, there was no suppression of the kindled amygdala focus. Allopregnanolone has anticonvulsant effects--and a good therapeutic index- against the secondarily generalized component of amygdala-kindled seizures. PMID- 17084971 TI - Embryonic zebrafish neuronal growth is not affected by an applied electric field in vitro. AB - Naturally occurring electric fields (EFs) have been implicated in cell guidance during embryonic development and adult wound healing. Embryonic Xenopus laevis neurons sprout preferentially towards the cathode, turn towards the cathode, and migrate faster towards the cathode in the presence of an external EF in vitro. A recent Phase 1 clinical trial has investigated the effects of oscillating EFs on human spinal cord regeneration. The purpose of this study was to investigate whether embryonic zebrafish neurons respond to an applied EF, and thus extend this research into another vertebrate system. Neural tubes of zebrafish embryos (16-17 somites) were dissected and dissociated neuroblasts were plated onto laminin-coated glass. A 100 mV/mm EF was applied to cell cultures for 4 or 20 h and the responses of neurons to the applied EFs were investigated. After 4h in an EF neurites were significantly shorter than control neurites. No other statistically significant effects were observed. After 20 h, control and EF exposed neurites were no different in length. No length difference was seen between cathodally- and anodally-sprouted neurites. Application of an EF did not affect the average number of neurons in a chamber. Growth cones did not migrate preferentially towards either pole of the EF and no asymmetry was seen in neurite sprout sites. We conclude that zebrafish neurons do not respond to a 100 mV/mm applied EF in vitro. This suggests that neurons of other vertebrate species may not respond to applied EFs in the same ways as Xenopus laevis neurons. PMID- 17084972 TI - A common A340T variant in PINK1 gene associated with late-onset Parkinson's disease in Chinese. AB - Parkinson's disease (PD) is a complex neurodegenerative disease with genetic risk factors. Common variants in genes implicated in hereditary forms of parkinsonism may be predisposing factors for sporadic PD. Recent studies have demonstrated that mutations in PINK1 (PARK6 locus) gene, encoding PTEN-induced kinase 1, are associated with both familial recessive and sporadic early onset parkinsonism. In order to assess whether the coding variant A340T contributes to the risk of late onset PD, we performed an association study of 539 PD patients with an onset age at or older than 50 and 525 controls in Chinese Han. Genotyping was performed by denaturing high performance liquid chromatography (DHPLC) combined with sequencing analyses. The A-allele frequency was 6.2% in PD and 4.2% in controls (p=0.0404), while G/A genotype frequencies were 12.4% in PD and 8.4% in the controls (p=0.0350). Our results yielded significant evidence for disease association between PINK1 A340T and PD with later onset (OR 1.55, 95% CI 1.04 2.32, p=0.0393), thus suggesting that PINK1 A340T variant may contribute to the risk for late-onset PD in Chinese. PMID- 17084973 TI - Atorvastatin slows down the deterioration of inner ear function with age in mice. AB - Statins have revolutionized the treatment of hypercholesterolemia due to their ability to inhibit cholesterol biosynthesis. Their immunomodulatory and anti inflammatory effects and positive effects on the treatment of atherosclerosis and its complications are well known. Here, we describe the effects of statins on the treatment of presbycusis in C57BL/6J mice. In this strain with accelerated aging, we demonstrate that animals treated with atorvastatin (10mg/kg per day in chow diet) for 2 months showed larger amplitudes of distortion product otoacoustic emissions (DPOAE) than did the non-treated control group. This finding indicates a better survival of outer hair cell function in the inner ear of C57BL/6J mice. The observed decreased expression of intercellular and vascular adhesion molecules in the aortic wall of atorvastatin-treated animals suggests that reducing endothelial inflammatory effects may contribute to the positive effect of atorvastatin on the amplitudes of DPOAE by influencing the blood supply to the inner ear. No such beneficial effect of statins was found in apoE(-/-) mice treated with atorvastatin under the same conditions. Our results suggest that statins could also slow down the age-related deterioration of hearing in man. PMID- 17084974 TI - Group I mGluR signaling in BSE-infected bovine-PrP transgenic mice. AB - Abnormalities of synapses and impaired synaptic transmission appear to be crucial in the pathogenesis of prion diseases. Excitotoxic mechanisms have been postulated as a major cause of neurodegeneration in these conditions. In this line, previous studies have shown abnormal group 1 metabotropic glutamate receptor signaling in Creutzfeldt-Jakob disease (CJD). In the present study, we have examined this pathway by western blotting in the cerebral cortex of bovine spongiform encephalopathy (BSE)-infected bovine-PrP transgenic mice at different days post-inoculation (dpi). Activation of post-synaptic metabotropic glutamate receptor 1 (mGluR1) promotes phospholipase Cbeta1 (PLCbeta1) activation which may activate, in turn, protein kinase C (PKC), which regulates gene expression. Densitometric analysis of the western blot bands revealed no differences in the protein levels of (mGluR1) through time, but demonstrated decreased levels of PLCbeta1 and protein kinase C delta (nPKCdelta) at 270dpi, at the time when mice showed neurological deficits accompanied by neuropathological changes and PrPres deposition in the brain. The present results show, for the first time impairment of the mGluR1/PLCbeta1/PKCdelta pathway signaling with disease-progression in a murine model of BSE. PMID- 17084975 TI - Behavioral and pharmacological description of oxaliplatin-induced painful neuropathy in rat. AB - We describe an animal model of nociceptive sensory neuropathy induced by repeat intravenous administration of oxaliplatin in which treated animals partly reproduce the characteristic pain symptoms in oxaliplatin-treated patients. We tested the ability of 1, 2 and 4 mg/kg oxaliplatin doses injected twice-weekly for four-and-a-half consecutive weeks to induce a nociceptive peripheral neuropathy in male Sprague-Dawley rats. The behavioral assessment revealed cold allodynia (10 degrees C) and hyperalgesia (4 degrees C) symptoms associated with a mechanical allodynia. The rats maintained a good general clinical status without motor dysfunction. The 2mg/kg oxaliplatin dose and the tail-immersion test in cold water (10 degrees C) were selected to compare pharmacological sensitivity between single administered drugs as morphine, lidocaine, carbamazepine, gabapentin and repeated administration of drugs as clomipramine, venlafaxine, calcium and magnesium solutions. Magnesium solution (90 mg/kg) and venlafaxine (7.5 mg/kg) administration induced an antinociceptive effect whereas gabapentin (300 mg/kg), clomipramine (2.5 mg/kg) and lidocaine (3 and 6 mg/kg) only induced an antiallodynic effect. PMID- 17084977 TI - The incidence of complex regional pain syndrome: a population-based study. AB - The complex regional pain syndrome (CRPS) is a painful disorder that can occur in an extremity after any type of injury, or even spontaneously. Data on the incidence of CRPS are scarce and mostly hospital based. Therefore the size of the problem and its burden on health care and society are unknown. The objective of the present study was to estimate the incidence of CRPS in the general population. A retrospective cohort study was conducted during 1996-2005 in the Integrated Primary Care Information (IPCI) project, a general practice research database with electronic patient record data from 600,000 patients throughout The Netherlands. Potential CRPS cases were identified by a sensitive search algorithm including synonyms and abbreviations for CRPS. Subsequently, cases were validated by electronic record review, supplemented with original specialist letters and information from an enquiry of general practitioners. The estimated overall incidence rate of CRPS was 26.2 per 100,000 person years (95% CI: 23.0-29.7). Females were affected at least three times more often than males (ratio: 3.4). The highest incidence occurred in females in the age category of 61-70 years. The upper extremity was affected more frequently than the lower extremity and a fracture was the most common precipitating event (44%). The observed incidence rate of CRPS is more as four times higher than the incidence rate observed in the only other population-based study, performed in Olmsted County, USA. Postmenopausal woman appeared to be at the highest risk for the development of CRPS. PMID- 17084978 TI - Catechol-O-methyltransferase inhibition increases pain sensitivity through activation of both beta2- and beta3-adrenergic receptors. AB - Catechol-O-methyltransferase (COMT), an enzyme that metabolizes catecholamines, has recently been implicated in the modulation of pain. Our group demonstrated that human genetic variants of COMT are predictive for the development of Temporomandibular Joint Disorder (TMJD) and are associated with heightened experimental pain sensitivity [Diatchenko, L, Slade, GD, Nackley, AG, Bhalang, K, Sigurdsson, A, Belfer, I, et al., Genetic basis for individual variations in pain perception and the development of a chronic pain condition, Hum Mol Genet 2005;14:135-43.]. Variants associated with heightened pain sensitivity produce lower COMT activity. Here we report the mechanisms underlying COMT-dependent pain sensitivity. To characterize the means whereby elevated catecholamine levels, resulting from reduced COMT activity, modulate heightened pain sensitivity, we administered a COMT inhibitor to rats and measured behavioral responsiveness to mechanical and thermal stimuli. We show that depressed COMT activity results in enhanced mechanical and thermal pain sensitivity. This phenomenon is completely blocked by the nonselective beta-adrenergic antagonist propranolol or by the combined administration of selective beta(2)- and beta(3)-adrenergic antagonists, while administration of beta(1)-adrenergic, alpha-adrenergic, or dopaminergic receptor antagonists fail to alter COMT-dependent pain sensitivity. These data provide the first direct evidence that low COMT activity leads to increased pain sensitivity via a beta(2/3)-adrenergic mechanism. These findings are of considerable clinical importance, suggesting that pain conditions resulting from low COMT activity and/or elevated catecholamine levels can be treated with pharmacological agents that block both beta(2)- and beta(3)-adrenergic receptors. PMID- 17084979 TI - Voltage-gated calcium channels and pain. PMID- 17084980 TI - The effect of age on the onset of pain interference in a general population of older adults: prospective findings from the North Staffordshire Osteoarthritis Project (NorStOP). AB - Pain that interferes with daily life appears to be strongly age-related in cross sectional studies, although the nature of this relationship over time has not been established. We have investigated the onset and persistence of pain and pain interference over a 3-year period to determine their association with age in older people. A 3-year follow-up postal survey was conducted of adults aged 50 years and over (n=5366) who had previously been recruited as part of the North Staffordshire Osteoarthritis Project. Four thousand two-hundred and thirty-four completed questionnaires were received (adjusted response 84.7%). The occurrence of pain interference at 3 years was 19.7% in persons free of such pain at baseline, higher in females than males (6.0% difference; 95% CI: 2.6%, 9.3%), and showed a clear age-related trend with a more than twofold increase from 50 to 59 years (16.0%) to the 80+ years (35%). Any pain at follow-up was reported by 48% of those pain-free at recruitment, and this figure was similar for males and females, and across 10-year age-groups. Persistence of pain interference (72.1%) at 3 years was high. In adults aged 50 years and over, the onset of pain that interferes with life shows a clear gender difference and a consistent rise with age into the oldest age-group. This was in strong contrast to the onset of pain which showed no gender or age-related trends. The implications for public health, as for the treatment of the individual, are twofold, relating to efforts to prevent disabling pain from occurring and to understand the factors that accelerate the impact which pain has on everyday life when people reach the oldest ages. PMID- 17084981 TI - Signalling via integrins: implications for cell survival and anticancer strategies. AB - Integrin-associated signalling renders cells more resistant to genotoxic anti cancer agents like ionizing radiation and chemotherapeutic substances, a phenomenon termed cell adhesion-mediated radioresistance/drug resistance (CAM-RR, CAM-DR). Integrins are heterodimeric cell-surface molecules that on one side link the actin cytoskeleton to the cell membrane and on the other side mediate cell matrix interactions. In addition to their structural functions, integrins mediate signalling from the extracellular space into the cell through integrin-associated signalling and adaptor molecules such as FAK (focal adhesion kinase), ILK (integrin-linked kinase), PINCH (particularly interesting new cysteine-histidine rich protein) and Nck2 (non-catalytic (region of) tyrosine kinase adaptor protein 2). Via these molecules, integrin signalling tightly and cooperatively interacts with receptor tyrosine kinase signalling to regulate survival, proliferation and cell shape as well as polarity, adhesion, migration and differentiation. In tumour cells of diverse origin like breast, colon or skin, the function and regulation of these molecules is partly disturbed and thus might contribute to the malignant phenotype and pre-existent and acquired multidrug resistance. These issues as well as a variety of therapeutic options envisioned to influence tumour cell growth, metastasis and resistance, including kinase inhibitors, anti integrin antibodies or RNA interference, will be summarized and discussed in this review. PMID- 17084982 TI - Aeromonas hydrophila and Aeromonas sobria bacteremia: rare pathogens of infection in a burn patient. PMID- 17084983 TI - Cocaine exposure in vitro induces apoptosis in fetal locus coeruleus neurons by altering the Bax/Bcl-2 ratio and through caspase-3 apoptotic signaling. AB - Cocaine inhibits survival and growth of rat locus coeruleus (LC) neurons, which may mediate alterations in attention, following in utero exposure to cocaine. These effects are most severe in early gestation during peak neuritogenesis. Prenatal cocaine exposure may specifically decrease LC survival through an apoptotic pathway involving caspases. Dissociated fetal LC neurons or substantia nigra (SN) neurons (control) were exposed in vitro to a pharmacologically active dose of cocaine hydrochloride (500 ng/ml) and assayed for apoptosis using terminal deoxynucleotidyl transferase mediated DNA nick end labeling and Hoechst methodologies. Cocaine exposure decreased survival and induced apoptosis in LC neurons, with no changes in survival of SN neurons. Activation of apoptotic signal transduction proteins was determined using enzyme assays and immunoblotting at 30 min, 1 h, 4 h and 24 h. In LC neurons, Bax levels were induced at 30 min and 1 h, following cocaine treatment, and Bcl-2 levels remained unchanged at all time points, altering the Bax/Bcl-2 ratio. The ratio was reversed for SN neurons (elevated Bcl-2 levels and transient reduction of Bax levels). Further, cocaine exposure significantly increased caspase-9 and caspase 3 activities at all time points, without changes in caspase-8 activity in LC neurons. In addition, cleavage of caspase-3 target proteins, alpha-fodrin and poly (ADP-ribose) polymerase (PARP) were observed following cocaine treatment. In contrast, SN neurons showed either significant reductions, or no significant changes, in caspase-3, -8 or -9 activities or caspase-3 target proteins, alpha fodrin and PARP. Thus, cocaine exposure in vitro may preferentially induce apoptosis in fetal LC neurons putatively regulated by Bax, via activation of caspases and their downstream target proteins. PMID- 17084984 TI - Stereological estimates of the basal forebrain cell population in the rat, including neurons containing choline acetyltransferase, glutamic acid decarboxylase or phosphate-activated glutaminase and colocalizing vesicular glutamate transporters. AB - The basal forebrain (BF) plays an important role in modulating cortical activity and influencing attention, learning and memory. These activities are fulfilled importantly yet not entirely by cholinergic neurons. Noncholinergic neurons also contribute and comprise GABAergic neurons and other possibly glutamatergic neurons. The aim of the present study was to estimate the total number of cells in the BF of the rat and the proportions of that total represented by cholinergic, GABAergic and glutamatergic neurons. For this purpose, cells were counted using unbiased stereological methods within the medial septum, diagonal band, magnocellular preoptic nucleus, substantia innominata and globus pallidus in sections stained for Nissl substance and/or the neurotransmitter enzymes, choline acetyltransferase (ChAT), glutamic acid decarboxylase (GAD) or phosphate activated glutaminase (PAG). In Nissl-stained sections, the total number of neurons in the BF was estimated as approximately 355,000 and the numbers of ChAT immuno-positive (+) as approximately 22,000, GAD+ approximately 119,000 and PAG+ approximately 316,000, corresponding to approximately 5%, approximately 35% and approximately 90% of the total. Thus, of the large population of BF neurons, only a small proportion has the capacity to synthesize acetylcholine (ACh), one third to synthesize GABA and the vast majority to synthesize glutamate (Glu). Moreover, through the presence of PAG, a proportion of ACh- and GABA-synthesizing neurons also has the capacity to synthesize Glu. In sections dual fluorescent immunostained for vesicular transporters, vesicular glutamate transporter (VGluT) 3 and not VGluT2 was present in the cell bodies of most PAG+ and ChAT+ and half the GAD+ cells. Given previous results showing that VGluT2 and not VGluT3 was present in BF axon terminals and not colocalized with VAChT or VGAT, we conclude that the BF cell population influences cortical and subcortical regions through neurons which release ACh, GABA or Glu from their terminals but which in part can also synthesize and release Glu from their soma or dendrites. PMID- 17084985 TI - Effects of histone deacetylation inhibition on neuronal differentiation of embryonic mouse neural stem cells. AB - Neural stem cells (NSCs) are multipotent cells that have the capacity for self renewal and for differentiation into the major cell types of the nervous system, i.e. neurons, astrocytes and oligodendrocytes. The molecular mechanisms regulating gene transcription resulting in NSC differentiation and cell lineage specification are slowly being unraveled. An important mechanism in transcriptional regulation is modulation of chromatin by histone acetylation and deacetylation, allowing or blocking the access of transcriptional factors to DNA sequences. The precise involvement of histone acetyltransferases and histone deacetylases (HDACs) in the differentiation of NSCs into mature functional neurons is still to be revealed. In this in vitro study we have investigated the effects of the HDAC inhibitor trichostatin A (TSA) on the differentiation pattern of embryonic mouse NSCs during culture in a minimal, serum-free medium, lacking any induction or growth factor. We demonstrated that under these basic conditions TSA treatment increased neuronal differentiation of the NSCs and decreased astrocyte differentiation. Most strikingly, electrophysiological recordings revealed that in our minimal culture system only TSA-treated NSC-derived neurons developed normal electrophysiological membrane properties characteristic for functional, i.e. excitable and firing, neurons. Furthermore, TSA-treated NSC derived neurons were characterized by an increased elongation and arborization of the dendrites. Our study shows that chromatin structure modulation by HDACs plays an important role in the transcriptional regulation of the neuronal differentiation of embryonic NSCs particularly as far as the development of functional properties are concerned. Manipulation of HDAC activity may be an important tool to generate specific neuronal populations from NSCs for transplantation purposes. PMID- 17084986 TI - Improvement of mouse brain mitochondrial function after deprenyl treatment. AB - Deprenyl is a selective monoamine oxidase (MAO) B inhibitor, widely used for treatment of Parkinson's disease. The present study shows that deprenyl treatment was able to improve mitochondrial function. Fourteen month old mice were injected i.p. with deprenyl (20 mg/kg) and killed 1.5 h after the administration. Different brain subcellular fractions were isolated from control and deprenyl treated animals to evaluate the effect of deprenyl on nitric oxide synthase (NOS) activity. Oxygen consumption, hydrogen peroxide (H(2)O(2)) production, mitochondrial membrane potential and calcium-induced permeability transition (MPT) were studied in intact mitochondria. In addition, the effect of deprenyl on respiratory complexes and MAO activities were evaluated in submitochondrial particles (SMP). Monoamine oxidase activity was found to be decreased by 55% in mitochondria from deprenyl-treated animals and as a consequence, H(2)O(2) production was significantly decreased. Deprenyl inhibited NOS activity in cytosolic fractions and SMP by 40% and 55%, respectively. In similar conditions, SMP from deprenyl-treated animals showed increased cytochrome oxidase activity. A 51% increase in the oxygen uptake in state 3 (active respiration state) was found after deprenyl treatment, but no significant changes were observed in state 4 (resting respiration state). Deprenyl treatment protected against calcium-induced depolarization and was able to inhibit calcium-induced MPT. This work provides evidence that deprenyl treatment exerts an improvement of brain mitochondrial function, through a reduction of free radical production, prevention of calcium induced MPT and maintaining a mitochondrial transmembrane potential. PMID- 17084987 TI - Immunocytochemical visualization of D-glutamate in the rat brain. AB - Using highly specific antisera directed against conjugated d-amino acids, the distribution of d-glutamate-, d-tryptophan-, d-cysteine-, d-tyrosine- and d methionine-immunoreactive structures in the rat brain was studied. Cell bodies containing d-glutamate, but not d-glutamate-immunoreactive fibers, were found. Perikarya containing this d-amino acid were only found in the mesencephalon and thalamus of the rat CNS. Thus, the highest density of cell bodies containing d glutamate was observed in the dorsal raphe nucleus, the ventral part of the mesencephalic central gray, the superior colliculus, above the posterior commissure, and in the subparafascicular thalamic nucleus. A moderate density of immunoreactive cell bodies was observed in the dorsal part of the mesencephalic central gray, above the rostral linear nucleus of the raphe, the nucleus of Darkschewitsch, and in the medial habenular nucleus, whereas a low density was found below the medial forebrain bundle and in the posterior thalamic nuclear group. Moreover, no immunoreactive fibers or cell bodies were visualized containing d-tryptophan, d-cysteine, d-tyrosine or d-methionine in the rat brain. The distribution of d-glutamate-immunoreactive cell bodies in the rat brain suggests that this d-amino acid could be involved in several physiological mechanisms. This work reports the first visualization and the morphological characteristics of conjugated d-glutamate-immunoreactive cell bodies in the rat CNS using an indirect immunoperoxidase technique. Our results suggest that the immunoreactive neurons observed have an uptake mechanism for d-glutamate. PMID- 17084988 TI - Human TLR11 gene is repressed due to its probable interaction with profilin expressed in human. PMID- 17084989 TI - Chronic dehydration may be a preventable risk factors for Alzheimer's disease. PMID- 17084990 TI - Squeezing the glans penis: a possible maneuver for improving the defecation process and preventing constipation. PMID- 17084991 TI - Dioscorea opposita reverses dexamethasone induced insulin resistance. AB - The effects of Dioscorea opposita (huai shan yao, HSY) on dexamethasone-induced insulin resistance were investigated in vitro and in vivo. D. opposita extract reduced significantly the blood insulin and glucose levels in dexamethasone induced diabetic rats. In vitro, HSY significantly enhanced insulin-stimulated glucose uptake in 3T3-L1 adipocytes. Moreover, HSY increase the mRNA expression of GLUT4 glucose transporter in 3T3-L1 adipocytes. These data suggest that D. opposita has insulin sensitivity that is associated with the regulation of GLUT4 expression. PMID- 17084992 TI - Tiliroside and gnaphaliin inhibit human low density lipoprotein oxidation. AB - Two flavonoids, gnaphaliin and tiliroside, isolated from Helichrysum italicum, were studied in vitro for their capacity to inhibit Cu(2+)-induced human low density lipoprotein (LDL) and diluted plasma oxidation. LDL oxidation was monitored by conjugated diene, thiobarbituric acid-reactive substances (TBARS) formation and electrophoretic mobility on agarose gel. Gnaphaliin and tiliroside increased the lag-phase for diene conjugate production in a dose-dependent manner. The reduction of TBARS production confirmed the antioxidant activity of gnaphaliin and tiliroside with 50% inhibitory concentration (IC(50)) values of 8.0+/-3.9 microM and 7.0+/-2.6 microM respectively. Furthermore, the flavonoids negated the Cu(2+)-induced increase in electrophoretic mobility of LDL. Antioxidant activity of gnaphaliin and tiliroside was significantly different when diluted plasma was oxidised by adding 1 mM CuSO(4). Although both flavonoids again reduced the TBARS production, tiliroside showed higher activity than gnaphaliin (IC(50)=10.6+/-2.5 microM vs. IC(50)>50 microM). In conclusion, tiliroside and gnaphaliin are antioxidants against in vitro Cu(2+)-induced LDL oxidation in the same order of magnitude compared to that of the reference drug, probucol. PMID- 17084993 TI - Contribution to the knowledge of the folk plant medicine in Calabria region (Southern Italy). AB - A series of research on plants used in Calabria (Southern Italy) in the folk plant medicine was carried out in the last twenty years. The use of 104 taxa distributed into 42 families are described. Among the major findings: Ceterach officinarum as hypotensive; Heliotropium europaeum in the case of urinary inflammations; Silybum marianum for haemorrhoids; Arundo donax for pertussis and cystitis; Allium cepa for sties and flu; Laurus nobilis as galactagogue; Passiflora incarnata for burns, haemorrhoids and skin inflammations; Scrophularia canina for rhagas and wounds; Urtica membranacea for tussis and tonsillitis. PMID- 17084994 TI - Psychometric properties of brief indexes designed to measure social-cognitive predictors of smoking initiation. AB - BACKGROUND: Constructing indexes which measure factors that may predict smoking initiation is essential for planning prevention programs. Our aim was to examine the criterion-related construct validity of brief psychological indexes of attitude, social influence and self-efficacy to be used in future studies for predicting smoking initiation in adolescents. METHODS: Five indexes were evaluated using cross-sectional data on 4819 adolescents age 13 in a random sample of schools in six Danish counties. Item analyses were performed with the graphical log-linear Rasch model, a modification of the Rasch model that allows for differential item functioning and positive local dependence. RESULTS: The three indexes social influence-norms, social influence-pressure and attitude showed acceptable deviations from the Rasch model, while the two indexes self efficacy and social influence-behavior were invalidated by negative local dependence. CONCLUSION: Of the five tested indexes, three (social influence norms, social influence-pressure and attitude) showed acceptable criterion related construct validity and may be considered unbiased representations of the theory-based factors if statistical analyses are appropriately adjusted. The results of this population-based study show that the graphical log-linear Rasch model is useful for evaluating health-related behavior indexes and identifying problems, which can be dealt with when using the indexes in future studies. PMID- 17084995 TI - Relationship between N-acetyl-aspartate in gray and white matter of abstinent methamphetamine abusers and their history of drug abuse: a proton magnetic resonance spectroscopy study. AB - OBJECTIVE: Altered concentrations of the brain metabolites, including N-acetyl aspartate (NAA) and myo-inositol (MI), may indicate neurotoxicity associated with drug abuse. In this study, the authors explored differences in brain metabolites between abstinent methamphetamine (MA) abusers and healthy comparison subjects and the associations between metabolite concentrations and clinical characteristics. METHOD: Proton magnetic resonance spectroscopy (MRS) was performed on 30 abstinent MA abusers and 20 healthy comparison subjects. Two sets of MA user subgroups were defined depending on abstinence duration (greater or less than 6 months) or the total cumulative MA dose (greater or less than 100 g lifetime). NAA and other metabolites were measured in the frontal gray and white matter and compared between MA abuser groups and healthy comparison subjects. RESULTS: MI concentrations were higher for the MA abusers relative to healthy comparison subjects. NAA concentration was lower in frontal white matter of MA abusers with a 'large' cumulative dose relative to those with a 'small' cumulative dose and to healthy comparison subjects. Additionally, in MA abusers NAA concentrations in frontal white matter correlated inversely with the cumulative MA dose. In contrast, there was no significant difference in frontal gray matter NAA concentration among the three groups. However, frontal gray matter NAA concentrations for MA abusers correlated negatively with the total cumulative MA dose and positively with the duration of abstinence. There were no differences between the different MA user groups for MI. CONCLUSIONS: The current findings suggest that MA-induced metabolic alterations of frontal gray and white matter are dose-dependent, for primarily male subjects. Additionally, these findings potentially suggest that the MA-related abnormalities may, in part, recover with abstinence in gray matter, but not in the white matter regions. PMID- 17084996 TI - Finding Nemo: molecular phylogeny and evolution of the unusual life style of anemonefish. AB - Anemonefish are a group of 28 species of coral reef fish belonging to the family Pomacentridae, subfamily Amphiprioninae, all characterized by living in symbiosis with sea anemones of several genera. Some anemonefish are specialized to cooperate with a single or few species of sea anemone, being immune to their poisonous tentacles but sensible to those of other species of sea anemones, while other anemonefish are more generalist and able to live together with a number of different species of sea anemone hosts. Despite the common life style, anemonefish species occur in a variety of colors, body shapes and degree of dependence from the host. To understand the evolutionary mechanisms responsible for the anemonefish diversification, we studied 23 out of 28 species of anemonefish by analyzing three mitochondrial regions: the cytochrome b gene, the 16S ribosomal RNA gene and the first half of the D-loop, a non-coding, regulatory region to reconstruct their molecular phylogeny through Bayesian and maximum parsimony approaches. The evolution of specialization was studied by means of character reconstruction methods. This work includes the highest number of anemonefish so far analyzed and particularly some species that had never been studied before. The results support a monophyletic origin for the subfamily Amphiprioninae, in contrast to the current taxonomy, based on morphological characters, that divides anemonefish into two separate genera. Moreover, we formulate some hypotheses concerning the life style and origin of the ancestral anemonefish. PMID- 17084997 TI - Ethanol oxidation into acetaldehyde by 16 recombinant human cytochrome P450 isoforms: role of CYP2C isoforms in human liver microsomes. AB - The involvement of cytochromes P450 (CYPs) in the oxidation of ethanol into acetaldehyde was investigated by using 16 recombinant human CYP isoforms. Apparent K(m) and V(m) were determined for CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2B6, CYP2C8, CYP2C9*1, CYP2C9*2, CYP2C9*3, CYP2C18, CYP2C19, CYP2D6, CYP2E1, CYP2J2, CYP3A4 and CYP4A11. All of the tested CYPs, except CYP2A6 and CYP2C18, metabolized ethanol into significant amounts of acetaldehyde and displayed K(m) values around 10mM. The significant correlation found between ethanol oxidation and CYP2E1, CYP3A4 and CYP1A2 catalytic activities in a panel of human liver microsomes confirmed the strong implication of these CYPs in ethanol metabolism. The contribution of CYP2C isoforms which are the most abundant in the liver after CYP3A4, was studied using selective inhibitors either with recombinant CYP2C isoforms or in human liver microsomes. Tienilic acid (100 microM) and ticlopidine (20 microM), mechanism-based inhibitors of CYP2C9 and CYP2C19, respectively, decreased ethanol oxidation by 8+/-1.2% and 7.6+/-1.6% in human liver microsomal samples while selective inhibitors of CYP2E1 (DEDTC 100 microM), CYP3A4 (TAO 50 microM) and CYP1A2 (furafylline 25 microM) decreased it by 11.9+/-2.1%, 19.8+/ 1.9% and 16.3+/-3.9%, respectively. As ethanol can be metabolized by most of CYPs, it helps to explain or predict alcohol-xenobiotics interactions which are of high importance in medical prescription. PMID- 17084998 TI - Intractable reflex audiogenic seizures in Aicardi syndrome. AB - Aicardi syndrome (AS) is a rare disorder which includes the triad of total or partial agenesis of the corpus callosum, infantile spasms, and chorioretinal anomalies. Seizures and electroencephalogram findings observed in AS are polymorphic with both focal and generalized seizures. We first report on a patient affected by AS who presented with reflex audiogenic seizures specifically triggered by the starting tune of a popular television news. No other type of stimuli, either simple or complex, were able to precipitate the seizures in the patient. The severe cortical-subcortical lesions commonly observed in AS are associated with hyperexcitability of the cortices and may well account for the broad electroclinical patterns noted in this group of patients. From our report, the context of these patterns should be extended to include reflex audiogenic seizures. PMID- 17084999 TI - Childhood autism and associated comorbidities. AB - Autism is a heterogeneous neurodevelopmental disorder with a variety of different etiologies, but with a heritability estimate of more than 90%. Although the strong correlation between autism and genetic factors has been long established, the exact genetic background of autism is still unclear. This review refers to all the genetic syndromes that have been described in children with pervasive developmental disorders (tuberous sclerosis, fragile X, Down, neurofibromatosis, Angelman, Prader-Willi, Gilles de la Tourette, Williams, etc.). Issues covered include prevalence and main characteristics of each syndrome, as well as the possible base of its association with autism in terms of contribution to the current knowledge on the etiology and genetic base of pervasive developmental disorders. PMID- 17085000 TI - Neurological presentation of Griscelli syndrome: obstructive hydrocephalus without haematological abnormalities or organomegaly. AB - Griscelli syndrome is a rare autosomal-recessive disorder characterised by partial albinism, immunodeficiency, organomegaly and accelerated phases. During accelerated phases, pancytopenia, haemophagocytosis, hypoproteinemeia occur which may be accompanied by neurological deterioration. Primary neurological presentation is rare and we report a case that presented with obstructive hydrocephalus and infiltrative lesions in the brain unaccompanied by other features of accelerated phase. Biopsy of these lesions demonstrated sinus histiocytosis. Electron microscopy of hair shaft and genetic studies established the diagnosis of Griscelli disease with RAB 27A mutation. PMID- 17085001 TI - Increased production of inflammatory cytokines in mild cognitive impairment. AB - Recent studies indicate that chronic inflammation plays a pathogenic role in both the central nervous system (CNS) and periphery in Alzheimer's disease (AD). We have screened for cytokines differentially produced by peripheral blood mononuclear cells (PBMCs) isolated from subjects with mild cognitive impairment (MCI) and mild AD subjects who had progressed from MCI using a commercially available cytokine array. Following determination of expressed cytokines, we quantified levels of the proinflammatory cytokines TNF-alpha, IL-6, and IL-8, and the anti-inflammatory cytokine IL-10 using flow cytometry. We have found a significant increase in the levels of IL-6, IL-8, and IL-10 produced by PBMCs stimulated for 24 h with phytohemagglutinin (PHA) in MCI subjects compared to healthy elderly controls. However, in PBMCs stimulated for 48 h with lipopolysaccharide (LPS), lower TNF-alpha/IL-10, IL-6/IL-10, and IL-8/IL-10 ratios were seen in MCI subjects. There were no differences in plasma levels of IL-8 between aged controls, MCI, and mild AD, and the levels of circulating IL-6 and IL-10 were below detection limits. Our data indicate that changes in cytokine production by PBMCs may be detected early in MCI, and an alteration of the immune response may precede clinical AD. PMID- 17085002 TI - Prostate cancer detection with 3-T MRI: comparison of diffusion-weighted and T2 weighted imaging. AB - OBJECTIVE: To compare the clinical value of diffusion-weighted (DW) and T2 weighted (T2W) imaging in detecting prostate cancer using a 3-Tesla (3T) magnetic resonance (MR) system. MATERIALS AND METHODS: Thirty-seven patients with suspected prostate cancer underwent T2W and DW imaging at 3T using an 8-channel phased-array coil. These images and apparent diffusion coefficient (ADC) maps were read retrospectively and blindly. The results were compared with histopathologic findings, and receiver operating characteristic (ROC) analysis was used to compare the cancer detection performance of T2W and DW imaging. RESULTS: The areas under the ROC curves for DW imaging and T2W imaging were 0.89 and 0.82, respectively. The performance of DW imaging in prostate cancer detection was significantly better than that of T2W imaging (P=0.0371). CONCLUSION: With a 3T MR system, the performance of DW imaging in detecting prostate cancer was better than that of T2W imaging. DW imaging appears to be a robust and reliable method to examine the whole prostate within an acceptable scan time in clinical settings. PMID- 17085003 TI - Self-expandable stent loaded with 125I seeds: feasibility and safety in a rabbit model. AB - OBJECTIVE: To evaluate technical feasibility and acute and subacute radiotolerance of a self-expandable stent loaded with (125)I seeds in the rabbit esophagus. METHODS: A self-expandable stent designed for esophageal application was made of 0.16mm nitinol wire and loaded with (125)I seeds (CIAE-6711). Twenty seven stents with three different radioactive dosages (n=9 in each dosage group) were implanted in the esophagus of healthy rabbits, while nine stents alone were used as controls. The stents were perorally deployed into the esophagus under fluoroscopic guidance. Radiological follow-up included plain chest film, CT scan, and barium esophagography which were undertaken in all rabbits of each group at 2, 4, and 8 weeks, respectively, which were correlated to histopathological findings. The stented esophageal segments along with their adjacent tissues were harvested for histopathological examinations. RESULTS: The stent was successfully deployed into the targeted esophageal segment in all rabbits. Neither (125)I seeds dislodged from the stent during the deployment, nor they did during the follow-up period. The greatest (16.2Gy) absorbed dose was found in the tissue 10mm from (125)I seeds at 8 weeks. Slight epithelial hyperplasia on the stent surface and submucosal inflammatory process developed at 2 weeks, which reached the peak at 8 weeks after the procedure. Significant thickness of the esophageal muscular layer was found at 8 weeks only in the groups with (125)I seeds. On radiologic follow-up, moderate strictures on both ends of the stents developed at 4 weeks and became severe at 8 weeks after the procedure in all groups. CONCLUSION: Deployment of a self-expandable stent loaded with (125)I seeds is technically feasible and safe within the first 8 weeks. Acute and subacute radiotolerance of the treated esophagus and its adjacent tissues by (125)I seeds is well preserved in a healthy rabbit model. PMID- 17085004 TI - Quantifying the usefulness of CT in evaluating seniors with abdominal pain. AB - OBJECTIVES: (1) Determine if older patients with abdominal pain who receive emergency department (ED) abdominal CT have changes in diagnosis and/or disposition more often than similar patients without CT; (2) compare physician confidence in diagnosis and disposition for patients with versus without CT; (3) document factors that most influence physician's decision to order abdominal CT in this population. METHODS: ED patients 60 years of age or older, with acute non traumatic abdominal pain were enrolled over a 6-week period. Physicians documented a preliminary and final ED diagnosis and disposition, along with pre- and post-evaluation confidence levels. Decision to order CT, along with clinical information most influencing that decision, was noted. Physician confidence levels and percent change in diagnosis and disposition were compared for patients with versus without CT. RESULTS: One hundred and twenty-six patients comprised study sample. Abdominal CT rate was 59% (95%CI, 50-67%). CT was associated with an increased change in diagnosis (46%; 95%CI, 4-58% versus 29%; 95%CI, 16-42%), but no change in disposition between patients with versus without CT. Preliminary diagnostic confidence was lower for EPs who ordered a CT than for those who did not (p<0.001). Patient history most influenced ordering CT, whereas prior lab/imaging results most influenced not ordering CT. CONCLUSION: Patients with CT had a change in diagnosis more often than those without. Preliminary diagnostic confidence was lower in CT group. Percent change in disposition did not differ between groups. Physicians most often ordered CT based on history and did not order CT when other diagnostic evaluation supported a specific diagnosis. PMID- 17085005 TI - Proteomic analysis identifies candidate proteins associated with distant recurrences in breast cancer after adjuvant chemotherapy. AB - Breast cancer is a heterogeneous disease and it is of importance to select patients with regard to different prognosis and treatment sensitivity to individualize treatment regimes. In this study we successfully adapted a protein extraction protocol from mRNA extracted tumor samples enabling two-dimensional gel electrophoresis (2-DE) analysis of samples previously analyzed by cDNA microarray. The aim was to find candidate proteins that distinguish breast cancer patients with or without recurrences after adjuvant CMF (cyclophosphamide, methotrexate and 5-FU) treatment within four years to follow-up. We identified several proteins distinguishing the recurrence group from the non-recurrence group, especially in the ER and PgR positive subgroup (n=7). The induced proteins were involved in translation/folding, iron ion binding, and protease inhibition, whereas proteins involved in signaling, ubiquitination, and splicing were decreased in expression. These results show that it is possible to use 2-DE to separate high abundant proteins in breast cancer tissue and to find discriminating proteins to identify patients with different prognosis after adjuvant CMF treatment. PMID- 17085006 TI - Patient participation in the medical specialist encounter: does physicians' patient-centred communication matter? AB - OBJECTIVE: Physicians' patient-centred communication is assumed to stimulate patients' active participation, thus leading to more effective and humane exchange in the medical consultation. We investigated the relationship between physicians' patient-centred communication and patient participation in a medical specialist setting. METHODS: Participants were 30 residents and specialists in internal medicine, and 323 of their patients. Participants completed a questionnaire prior to a (videotaped) follow-up consultation. Physicians' patient centred communication was assessed by coding behaviours that facilitate or rather inhibit patients to express their perspective. Patient participation was determined by assessing (a) their relative contribution to the conversation, and (b) their active participation behaviour. Analyses accounted for relevant background characteristics. RESULTS: Physicians' facilitating behaviour was found to be positively associated with patients' relative contribution to the conversation as well as patients' active participation behaviour. Physicians' inhibiting behaviour was not related to patients' relative contribution, and was, unexpectedly, positively associated with patients' active participation behaviour. Physicians' behaviour was particularly associated with patients' expression of concerns and cues. CONCLUSIONS: Physicians in internal specialist medicine appear to be able to facilitate patients' active participation in the visit. The findings indicate that inhibiting behaviour may not have the expected blocking effect on patient participation: patients voiced their perspectives just the same and expressed even more concerns. Showing inhibiting behaviour may, alternatively, be a physician's response to the patient's increased participation in the encounter. PRACTICE IMPLICATIONS: The results may give directions for future medical education and specialist training. PMID- 17085007 TI - Recurrence rates after DCE-MRI image guided planning for breast-conserving surgery following neoadjuvant chemotherapy for locally advanced breast cancer patients. AB - BACKGROUND: Neoadjuvant therapy results in a significant increase in breast conserving surgery. However, traditional imaging methods are unable to accurately predict the extent of viable residual disease leading to uncertainty in surgical planning and some previous studies have shown a disproportionately high incidence of locoregional recurrence. Dynamic contrast enhanced-MRI (DCE-MRI) has been shown to provide a potentially more accurate prediction of residual disease. RESULTS: Patients undergoing neoadjuvant chemotherapy for breast cancer in our unit are staged with the DCE-MRI of the breast performed at 1.5 T before, during and after treatment and the final result was used to plan surgery. Two hundred and four patients with breast cancer were treated with neoadjuvant chemotherapy between 1996 and April 2005. Eighteen of these patients had distant metastases at the time of initial diagnosis and so were excluded from the present study. Following neoadjuvant chemotherapy, 186 patients underwent surgical treatment. Of these, 68 patients had breast-conserving surgery. At a median follow up of 30 months (range: 5.6-72 months) 21 patients in this group developed subsequent recurrence (21/68 - 30%) of whom 9 (9/68 - 13%) had locoregional recurrence, 7 had local recurrence (7/68 - 10%), and 17 (17/68 - 25%) had distant recurrence. Logistic regression analysis revealed only vascular invasion (p=0.006) of the tumour to be significantly associated with overall recurrence. None of the pathological factors (ER, PR status, vascular invasion, lymph node metastases, pathological complete response to neoadjuvant chemotherapy) showed a significant association with locoregional recurrence. CONCLUSION: Breast-conserving surgery with DCE-MRI planning after neoadjuvant chemotherapy provides an acceptable level of local recurrence without the need for mastectomy. PMID- 17085008 TI - Carcinoma showing thymus-like element (CASTLE) of thyroid: combined modality treatment in 3 patients with locally advanced disease. AB - OBJECTIVES: To examine the value of chemotherapy, radiotherapy and surgery for treatment of locally advanced carcinoma showing thymus-like element (CASTLE) of the thyroid. METHODS: Retrospective analysis of 3 Chinese patients in a tertiary referral center in Hong Kong. RESULTS: CASTLE is a rare thyroid malignancy with a frequency of only 0.15% (3/2033 patients) in our center. Three patients (M:F=2:1) aged 43, 49 and 62 years were studied. All 3 patients had advanced T4 disease with extensive tracheal infiltration and airway compression. None had lymph node or distant metastasis. Total thyroidectomy, combined with chemotherapy and radiotherapy, was effective in local control and symptom relief. Etoposide and carboplatin were tried in 2 patients with positive response. Neoadjuvant chemotherapy shrank the tumor rapidly and relieved symptoms of airway compression. All 3 patients had external radiotherapy resulting in good local control. In a patient with inoperable disease, chemotherapy and radiotherapy rendered the disease operable. All 3 patients were symptom-free and alive at 6, 2.5 and 1.8 years after diagnosis. CONCLUSIONS: CASTLE is locally infiltrative and presents at advanced T stage in this small series. Chemotherapy and radiotherapy, apart from surgery, are effective treatment modalities. In cases of inoperable disease or advanced local disease, they can be employed in combination with surgery. Organ preservation of larynx and trachea may be achieved. Chemotherapy can be very useful for rapid relief of symptoms, especially in shrinking tumor to prevent airway obstruction. PMID- 17085009 TI - [Effects of music therapy in intensive care unit without sedation in weaning patients versus non-ventilated patients]. AB - OBJECTIVES: Music has been found to be an effective nonpharmacologic adjunct for managing anxiety and promoting relaxation in limited trials of critically ill patients. However, its effects have not been compared in intubated patients during weaning from mechanical ventilation with non-intubated patients spontaneously breathing. STUDY DESIGN: A cross-over randomized experimental design. PATIENTS AND METHODS: Thirty patients were studied (intubated group n = 15, non-intubated group n = 15). Patients were randomized to receive either 20 minutes of uninterrupted rest or then 20 minutes of music therapy or the music therapy first and then the uninterrupted rest period. Patients selected a relaxing music of their choice from a selection including different types of music. Heart rate (HR), systolic blood pressure (SAP), respiratory rate (RR) and bispectral index (BIS score) were recorded each 5-min intervals throughout both periods (rest and music). Agitation/sedation state and pain were evaluated by the Richmond-Agitation-Sedation-Scale (RASS) and the Numerical-Rating-Scale (NRS) respectively, before and after each studied periods. Music have not been performed in five patients (5/35 = 14%). RESULTS: Music significantly decreased HR (88+/-15 vs 82+/-15, P<0.05), SAP (137+/-17 vs 128+/-14, P<0.05), RR (25+/-3 vs 22+/-4, P<0.05), BIS (94+/-5 vs 81+/-10, P<0.01), RASS (+0.1+/-0.7 vs -0.7+/ 0.9, P<0.05) and NRS (4.4+/-1.7 vs 1.9+/-1.3, P<0.01) in both intubated and non intubated groups whereas no significant change was observed during the rest period. The variations level studied parameters induced by music were comparable for the two groups. CONCLUSION: A single music therapy session was found to be effective for decreasing anxiety and promoting relaxation, as indicated by decreases in heart rate, blood pressure, BIS and respiratory rate over the intervention period in intubated patients during weaning phase. PMID- 17085010 TI - Development and quality assurance of computer-based assessment batteries. AB - The purpose of this article is to outline critical elements in the development and quality assurance (QA) assessment of a computer-based assessment battery (CAB). The first section of the article provides an overview of the life cycle of a representative CAB, typical evolutionary stages, and many of the essential considerations for designing and developing a CAB. The second section of the article presents a model for conducting a quality assurance assessment of a CAB. A general narrative of several steps in the QA process is supported by a table of recommended QA assessment elements. Although this QA process model may not be definitive for all cases, it provides a general framework within which a systematic assessment of any CAB can be conducted. PMID- 17085011 TI - PTSD: a problematic diagnostic category. AB - Since the publication of Diagnostic and Statistical Manual of Mental Disorders (DSM)-III, posttraumatic stress disorder (PTSD) has become a remarkably dominant theme in mental health discourse and diagnostic practice. This development has been encouraged by the diagnosis being officially presumed to exist in acute, chronic, delayed, complex, subdromal, and even "masked" forms. Here, we present an historical and clinical review that indicates how, since 1980, the term PTSD (along with its dubious embellishments) replaced established views on mental responses to trauma to the detriment of patient care and psychiatric investigation. From this historical perspective, we review and evaluate the natural course of emotional and behavioral reactions to traumatic experiences, and as well their assessment, formulation, and therapeutic management in both civilian and military situations. From this we conclude that the concept of PTSD has moved the mental health field away from, rather than towards a better understanding of the natural psychological responses to trauma. A return to prior standards of diagnostic practice and therapeutic planning would greatly benefit patient care, rehabilitative services to veterans, and epidemiologic research. PMID- 17085012 TI - An integrated map of p53-binding sites and histone modification in the human ENCODE regions. AB - TP53 (tumor protein p53; p53) regulates its target genes under various cellular stresses. By combining chromatin immunoprecipitation with oligonucleotide microarrays, we have mapped binding sites of p53 (p53-BS) in the genome of HCT116 human colon carcinoma cells, along with those of acetylated H3, acetylated H4, and methylated H3-K4. We analyzed a 30-Mb portion of the human genome selected as a representative model by the ENCODE Consortium. In the region, we found 37 p53 BS, of which the p53-binding motif was present in 32 (86%). Acetylated histone H3 and H4 were detected at 14 (38%) and 33 (89%) of the p53-BS, respectively. A significant portion (58%) of H4 acetylation in the p53-BS was not accompanied by H3 acetylation. Acetyl H3 were preferentially located at the 5' and 3' ends of genes, whereas acetyl H4 were distributed widely across the genome. These results provide novel insights into how p53 binding coordinates with histone modification in human. PMID- 17085013 TI - 3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitor Atorvastatin mediated effects depend on the activation status of target cells in PLP-EAE. AB - The effect of Atorvastatin on transcriptional activity in murine experimental autoimmune encephalomyelitis (EAE) induced by PLP peptide 139-151 was analyzed by DNA microarray technique in lymph nodes and spinal cord at onset (10 days), height (20 days) and first remission (30 days) of disease. Fourteen genes were selectively influenced by Atorvastatin in EAE mice. They are mainly related to immune cell functions and regulation of cell-to-cell interaction. Interestingly, seven genes were also differentially regulated in CFA-injected control mice. But qualitative and quantitative differences to EAE mice argue for a dependency of statin effects on the activation status of target cells. Differential regulation of the newly detected candidate genes of statin effects COX-1 and HSP-105 and the previously known statin-responsive genes ICAM-1 and CD86 was confirmed by Western blot and immunohistochemistry. Flow cytometric analysis of lymph node cells revealed that the effect of Atorvastatin treatment in non-immunized healthy animals resembled the effect of immunization with PLP peptide regarding changes of T helper cells, activated B cells and macrophages. In EAE mice, these effects were partially reversed by Atorvastatin treatment. Monitoring of expression of the newly identified candidate genes and patterns of lymphocyte subpopulations might predict the responsiveness of multiple sclerosis patients to statin treatment. PMID- 17085014 TI - STAT1 as a key modulator of cell death. AB - Signal transducers and activators of transcription (STATs) are latent cytoplasmic transcription factors that mediate various biological responses, including cell proliferation, survival, apoptosis, and differentiation. Among the members of the STAT family, accumulating evidence now indicates an important role for STAT1 in various forms of cell death. Depending upon stimuli or cell types, STAT1 can modulate a broad spectrum of cell death, comprising both apoptotic and non apoptotic pathways. STAT1-dependent regulation of cell death is largely dependent on a transcriptional mechanism such as the activation of death-promoting genes. However, non-transcriptional mechanisms such as STAT1 interaction with TRADD, p53, or HDAC have been implicated in the regulation of cell death by STAT1. Furthermore, STAT1 itself is also subject to complex forms of regulation such as post-translational protein modification, which can critically affect STAT1 signaling and STAT1-dependent cell death. Given the reports showing that dysregulation of STAT1 signaling is associated with various pathological conditions, including the development of cancer, a better understanding of the mechanism underlying STAT1 regulation of cell death may lead to successful strategies for targeting STAT1 in such pathological settings. PMID- 17085015 TI - The involvement of nitric oxide in the anticonvulsant effects of alpha-tocopherol on penicillin-induced epileptiform activity in rats. AB - A variety of animal seizure models exist which help to document the effects of alpha-tocopherol (Vitamin E) and specify its action. In the present study, we provide further evidence for the functional involvement of NO in the anticonvulsant effects of alpha-tocopherol on penicillin-induced epileptiform electrocorticographical (ECoG) activity in rats. The epileptiform ECoG activity was induced by microinjection of penicillin into the left sensorimotor cortex. Thirty minutes after penicillin injection, the most effective dose of alpha tocopherol (500 mg/kg) was administrated intramuscularly (i.m.). Alpha-tocopherol decreased the frequency of penicillin-induced epileptiform ECoG activity without changing the amplitude. The effect of systemic administration of nitric oxide synthase (NOS) inhibitor, NG-nitro-L-arginine methyl ester (L-NAME) and NO substrates, L-arginine and sodium nitro prusside (SNP) on anticonvulsive effects of alpha-tocopherol was investigated. The administration of L-NAME (60 mg/kg, i.p.) did not influence the frequency of epileptiform ECoG activity while administration of L-arginine (500 mg/kg, i.p.) and SNP (6 mg/kg, i.p.) significantly decreased in the penicillin-treated group. The administration of L NAME (60 mg/kg, i.p.) 10 min after alpha-tocopherol (500 mg/kg, i.m.) application reversed the anticonvulsant effects of alpha-tocopherol. The administration of L arginine (500 mg/kg, i.p.) and SNP (6 mg/kg, i.p.) did not affect the frequency of epileptiform ECoG activity in alpha-tocopherol supplemented group. L-arginine and SNP did not provide an additional anticonvulsant effect in alpha-tocopherol supplemented group. These results support the involvement of the nitric oxide pathway in the anticonvulsant effect of alpha-tocopherol on the penicillin induced epileptiform ECoG activity. PMID- 17085016 TI - Seizure prediction: the impact of long prediction horizons. AB - Several procedures have been proposed to be capable of predicting the occurrence of epileptic seizures. Up to now, all proposed algorithms are far from being sufficient for a clinical application. This is, however, often not obvious when results of seizure prediction performance are reported. Here, we discuss impacts of long prediction horizons with respect to clinical needs and the strain on patients by analyzing long-term continuous intracranial electroencephalography data. PMID- 17085017 TI - Administration of levetiracetam after prolonged status epilepticus does not protect from mitochondrial dysfunction in a rodent model. AB - Neuronal death and dysfunction occur after status epilepticus (SE), and is associated with mitochondrial enzyme damage. We previously showed, using the rat perforant pathway stimulation model, that levetiracetam administration (LEV; 1000 mg/kg intraperitoneal) during established SE reduces seizure severity and prevents mitochondrial dysfunction. We now show that administration of the same dose of LEV after 5h SE, does not protect from mitochondrial dysfunction. PMID- 17085018 TI - White matter abnormalities and brain activation in schizophrenia: a combined DTI and fMRI study. AB - Diffusion tensor imaging (DTI) studies of schizophrenia have revealed white matter abnormalities in several areas of the brain. The functional impact on either psychopathology or cognition remains, however, poorly understood. Here we analysed both functional MRI (during a working memory task) and DTI data sets in 18 patients with schizophrenia and 18 controls. Firstly, DTI analyses revealed reductions of fractional anisotropy (FA) in the right medial temporal lobe adjacent to the right parahippocampal gyrus, likely to contain fibres of the inferior cingulum bundle, and in the right frontal lobe. Secondly, functional MRI revealed prefrontal, superior parietal and occipital relative hypoactivation in patients with the main effect of task. This was accounted for by reduced prefrontal activation during the encoding phase of the task, but not during maintenance or retrieval phases. Thirdly, we found a direct correlation in patients between the frontal FA reduction (but not medial temporal reductions) and fMRI activation in regions in the prefrontal and occipital cortex. Our study combining fMRI and DTI thus demonstrates altered structure-function relationships in schizophrenia. It highlights a potential relationship between anatomical changes in a frontal-temporal anatomical circuit and functional alterations in the prefrontal cortex. PMID- 17085019 TI - Evidence of excessive concentrations of 5-flucytosine in children aged below 12 years: a 12-year review of serum concentrations from a UK clinical assay reference laboratory. AB - 5-flucytosine (5-FC) is an antifungal drug used for the treatment of serious infections caused by Candida or Cryptococcus spp. In the UK, the recommended pre- and post-dose serum therapeutic ranges are 30-40 mg/L and 70-80 mg/L, respectively. A 12-year retrospective review of serum concentrations of 5-FC in three groups of children aged 1-30 days (n=167), 31-60 days (n=102) and 91 days to 12 years (n=122) was conducted. In these three age groups, 65.1%, 44.4% and 21.3% of pre-dose samples and 39.3%, 29.2% and 19.7% of post-dose samples were above the recommended ranges. Both the mean concentration and the percentage of concentrations above the recommended ranges were significantly higher in the youngest age group (1-30 days old), suggesting that the standard dose of 100 mg/kg daily may not be an appropriate dose in this age group. PMID- 17085020 TI - Regional brain serotonin and dopamine transporter binding capacity in suicide attempters relate to impulsiveness and mental energy. AB - To study different aspects of regional pre-synaptic brain (123)I-beta-CIT uptake on serotonin and dopamine re-uptake sites in drug-free suicide attempters in comparison with age- and sex matched control subjects, single photon emission computed tomography (SPECT) measurements were analysed for regional serotonin re uptake (5HTT) and dopamine re-uptake (DAT) capacity (binding potential, BP()) after i.v. (123)I-beta-CIT administration. All suicide attempters were examined concerning seriousness of the attempt, and DSM-IV diagnosis. Both suicide attempters and control subjects were tested for psychotropic drugs, and completed the Marke-Nyman Temperament (MNT) test, including solidity (level of impulsiveness/initiative) and validity (level of mental energy). We found no significant difference between suicide attempters and control subjects concerning the regional levels of 5HTT BP() or DAT BP(). However, in suicide attempters, but not controls, we found significant regional correlations between MNT variables and SPECT results. We interpret the discrepant findings in suicide attempters and control subjects to be due to a disability of the suicide attempters to regulate their serotonin and dopamine levels, e.g. in response to external stress. PMID- 17085021 TI - Emotional memory in depersonalization disorder: a functional MRI study. AB - This study examines emotional memory effects in primary depersonalization disorder (DPD). A core complaint of DPD sufferers is the dulling of emotional responses, and previous work has shown that, in response to aversive stimuli, DPD patients do not show activation of brain regions involved in normal emotional processing. We hypothesized that DPD sufferers would not show the normal emotional enhancement of memory, and that they would not show activation of brain regions concerned with emotional processing during encoding and recognition of emotional verbal material. Using fMRI, 10 DPD patients were compared with an age matched healthy control group while performing a test of emotional verbal memory, comprising one encoding and two recognition memory tasks. DPD patients showed significantly enhanced recognition for overtly emotive words, but did not show enhancement of memory for neutral words encoded in an emotive context. In addition, patients did not show activation of emotional processing areas during encoding, and exhibited no substantial difference in their neural responses to emotional and neutral material in the encoding and emotional word recognition tasks. This study provides further evidence that patients with DPD do not process emotionally salient material in the same way as healthy controls, in accordance with their subjective descriptions of reduced or absent emotional responses. PMID- 17085022 TI - A PET study of 5-HT1A receptors at different phases of the menstrual cycle in women with premenstrual dysphoria. AB - The cause of premenstrual dysphoric disorder (PMDD) is largely unknown. It has been hypothesized that normal ovarian function triggers PMDD-related biochemical events within the brain and that serotonin plays an important role. In the present study, positron emission tomography (PET) and [carbonyl-(11)C]WAY-100635 were used to examine serotonin 5-HT(1A) receptors in a control group of women and in a group of women with PMDD. Two PET examinations were performed in each subject, one before (follicular phase) and one after ovulation (luteal phase). Each subject's menstrual cycle was confirmed by ultrasonography of the ovaries as well as with hormone levels in blood and urine. The 5-HT(1A) binding potential was measured in six regions of interest and calculated according to the simplified reference tissue model. In the raphe nuclei, the 5-HT(1A) binding potential changed from the follicular to the luteal phase of the menstrual cycle in asymptomatic controls. In women with PMDD, the observed change between phases was significantly smaller. The results are in concordance with previously reported challenge studies of 5-HT(1A) receptor-mediated effects indicating different serotonergic responses between women with PMDD and controls. The study principally provides new support, in vivo, for a serotonergic dysregulation in women with PMDD. PMID- 17085023 TI - Temporal and spatial dynamics of blocking and ripening effects on bacterial transport through a porous system: a possible explanation for CFT deviation. AB - The studies on transport of particles across porous systems are based on the Colloid Filtration Theory (CFT). According to CFT, the collision efficiency is constant along the system length [J.N. Ryan, M. Elimelech, Colloids Surf. A: Physicochem. Eng. Aspects 107 (1996) 1-56]. Decreasing values of collision efficiency have been reported, a phenomenon that has been interpreted as a deviation from the CFT [X. Li, T.D. Scheibe, W.P. Johnson, Environ. Sci. Technol. 38 (2004) 5616-5625; N. Tufenkji, J.A. Redman, M. Elimelech, Environ. Sci. Technol. 37 (2003) 616-623; N. Tufenkji, M. Elimelech, Langmuir 20 (2004) 10818 10828; N. Tufenkji, M. Elimelech, Langmuir 21 (2005) 841-852]. This paper presents data on transport of Bacillus megaterium spores through quartz sand columns. The occurrence of consecutive phases of increase and decrease of the values of C/C(0), the effluent spore concentration expressed as a fraction of the influent spore concentration, is reported. These patterns of change in C/C(0) were interpreted as the result of the concomitant occurrence of blocking and ripening, the prevalence of these phenomena in different moments of the experiment, and the spatial distribution of the prevalence of blocking and ripening effects along the porous system. It is argued that this spatial distribution in the predominance of blocking and ripening, what leads to the intensification of ripening at the entrance of the porous system, might be a possible explanation for the reported deviation from the CFT for experimental conditions where ripening and blocking take place. PMID- 17085024 TI - A novel dissolution method relevant to intestinal release behaviour and its application in the evaluation of modified release mesalazine products. AB - Mesalazine (5-ASA) is a compound being used in the therapy of inflammatory bowel disease (IBD). Considering the fact that 5-ASA is locally active and that the location of inflammation in IBD may vary, it is recognized that the release profile of 5-ASA drugs is the dominant factor for adequate local bioavailability. Furthermore, it is hypothesized that systemic absorption of 5-ASA (mainly in the upper intestinal segments) increases the risk of side effects. These facts relate to the conclusion that a method determining the dissolution profile under biorelevant conditions is a valuable tool for evaluation and comparison of 5-ASA products. We tested several commercially available products (Salofalk tablets, Salofalk granules, Asacol tablets, Pentasa tablets and granules) in a gastro intestinal simulation system (GISS). The GISS is based on the pharmacopeial dissolution test. The release profiles of all products are in agreement with their technological concepts. The percentage of the dose released in the simulated colon is small in all products. The GISS is a robust system able to discriminate between products which apply different modified-release technologies. Colon-selectivity of modified-release 5-ASA products might further be improved. The commercially available 5-ASA containing oral dosage forms exhibit different release profiles, which suggests that the optimal product may differ per patient. PMID- 17085025 TI - Cellulose acetate butyrate-pH/thermosensitive polymer microcapsules containing aminated poly(vinyl alcohol) microspheres for oral administration of DNA. AB - The aim of this work is to safely transport bioadhesive microspheres loaded with DNA to intestine and to test their bioadhesive properties. Poly(vinyl alcohol) (PVA) microspheres were prepared by dispersion reticulation with glutaraldehyde and further aminated. These microspheres were firstly loaded with plasmid DNA by electrostatic interactions and then entrapped in cellulose acetate butyrate (CAB) microcapsules for gastric protection. The entrapped PVA microspheres do not have enough force by swelling to produce the rupture of CAB shell, therefore the resistance of microcapsules was weakened by incorporating different amount of the pH/thermosensitive polymer (SP) based on poly(N-isopropylacrylamide-co-methyl methacrylate-co-methacrylic acid) (NIPAAm-co-MM-co-MA). This polymer is insoluble in gastric juice at pH 1.2 and 37 degrees C, but quickly solubilized in intestinal fluids (pH 6.8 and pH 7.4). Therefore, DNA loaded PVA microspheres were not expelled in acidic media but were almost entirely discharged in small intestine or colon. The integrity of DNA after entrapment was tested by agarose gel electrophoresis indicating that no DNA degradation occurs during encapsulation. The percentage of adhered microspheres on the mucus surface of everted intestinal tissue was 65+/-18% for aminated PVA microspheres without DNA and almost 50+/-15% for those loaded with DNA. Non-aminated PVA microspheres display the lowest adhesive properties (33+/-12%). In conclusion DNA loaded microspheres were progressively discharged in intestine. The integrity of DNA was not modified after entrapment and release, as proved by agarose gel electrophoresis. Both loaded and un-loaded aminated microspheres display good bioadhesive properties. PMID- 17085026 TI - A novel keratinase from Clostridium sporogenes bv. pennavorans bv. nov., a thermotolerant organism isolated from solfataric muds. AB - A bacterium which can grow on chicken feathers and which exhibits keratinolytic activity was isolated from solfataric muds. It was classified as belonging to the genus Clostridium and closely related to C. sporogenes. Based on its unique capability to degrade chicken feathers, it was designated as Clostridium sporogenes bv. pennavorans bv. nov. The keratinase purified from the culture supernatant is a monomer of 28.7kDa molecular mass. The enzyme is relatively thermostable and is active over a broad range of temperature and pH. Specific enzymatic assays demonstrate that keratinase can act on a large variety of soluble and insoluble protein substrates. PMID- 17085027 TI - Behavioral and biochemical studies of total furocoumarins from seeds of Psoralea corylifolia in the chronic mild stress model of depression in mice. AB - Depression is related to alterations of the monoamine oxidase (MAO), hypothalamic pituitary-adrenal (HPA) axis, and oxidative systems, and some antidepressants achieve their therapeutic effects through alteration of following biochemical markers of depression: MAO-A and MAO-B activities, cortisol levels, superoxide dismutase (SOD) activity and malondialdehyde (MDA) levels. The seeds of Psoralea corylifolia, otherwise known as Buguzhi, have long been used for treatments of various symptoms associated with aging in China. Furocoumarins are the most widespread secondary metabolites in this species. The present study was designed to evaluate the potential antidepressant-like activity of total furocoumarins of P. corylifolia (TFPC) in the chronic mild stress (CMS) model of depression. Mice subjected to CMS exhibited a reduction in sucrose intake. Conversely, brain MAO-A and MAO-B activities, plasma cortisol levels, and liver SOD activity and MDA levels were increased following CMS exposures. The time-course for reversal of CMS-induced deficits in sucrose consumption by TFPC was dose-dependent. Thus, the statistically significant effect of the higher dose of TFPC (50 mg/kg body wt.) was observed after 3 days of treatment, while 6 days of treatment were required in the group receiving a lower dose (30 mg/kg body wt.) of TFPC. TFPC reversed these biochemical changes. These results suggest that TFPC may possess potent and rapid antidepressant properties that are mediated via MAO, the HPA axis and oxidative systems and these antidepressant actions could make TFPC a potentially valuable drug for the treatment of depression in the elderly. PMID- 17085028 TI - Putrescine-1,4-dicinnamide from Pholiota spumosa (Basidiomycetes) inhibits cell growth of human prostate cancer cells. AB - Previously, it was isolated from the fruiting bodies of the gilled mushroom Pholiota spumosa (Basidiomycetes, Strophariaceae), putrescine-1,4-dicinnamide, a phenylpropanoid derivative conjugated with polyamine putrescine never isolated before as a natural compound. Recently, polyamine analogs that are similar in structure to the natural polyamines but that cannot mimic their functions that are essential for cellular growth and differentiation, have shown antitumor activity in several types of human cancer cells. Therefore, we have now investigated the response of DU-145 cells, a well characterized androgen independent human prostate cancer (PCA) cell line, to this phenylpropanoid derivative. The results presented here demonstrate that putrescine-1,4 dicinnamide, as suggested for polyamine analogs synthesized artificially, inhibits the cell growth of cancer cells inducing apoptosis cell death, mediated, at least in part, by the activation of caspase cascades, that at higher doses shift to necrosis, through the increase of reactive oxygen species (ROS) generation. PMID- 17085029 TI - Down-regulation of prostate specific antigen in LNCaP cells by flavonoids from the pollen of Brassica napus L. AB - The pollen of Brassica napus L. has been used in China to treat benign prostatic hyperplasia (BPH) for over decades. In this study, the pollen of Brassica napus L. was extracted successively with chloroform, ethyl acetate and ethanol. The ethyl acetate extract showed strong activity in decreasing the secretion of prostate specific antigen (PSA) in LNCaP cells as compared to two other extracts, measured by ELISA with finasteride as positive control in the assay. Five flavonoids were subsequently isolated from the active extract using bioassay guided fractionation. They were Naringenin (1); Luteolin (2); Kaempferol (3); Kaempferol 3-(3-E-p-coumaroyl-alpha-L-rhamnopyranoside) (4); and Kaempferol 3 (2,3-di-E-p-coumaroyl-alpha-L-rhamnopyranoside) (5). All these compounds inhibited PSA secretion significantly, with IC50 values in the range of 5-50 microM. Compounds 2, 4 and 5 showed moderate cytotoxicity to LNCaP cells within the active concentration range, while compounds 1 and 3 showed no cytotoxicity. Further studies on the mechanism action of these compounds were performed by evaluating their activation of estrogen receptor (ER) and antagonistic activities on androgen receptor (AR) in cell-based reporter gene assays. All compounds described here were first isolated from the pollen of Brassica napus L. PMID- 17085030 TI - Walking distance is a predictor of exacerbations in patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Patients with chronic obstructive pulmonary disease (COPD) are responsible for a high utilisation of the health care resources, and the cost is expected to increase. Physiological measures of lung function often fail to describe the impact the symptoms have on exacerbations, days of hospitalisation, and on a patient's health. METHODS: Twenty-one patients (14 female) with COPD (65 years, 40-79 years) admitted to the Department of Respiratory Medicine in Uppsala, performed a pulmonary function test (FEV(1)% predicted=37) and health status measurement (St. Georges Respiratory Questionnaire, SGRQ) at discharge. Four to six weeks after discharge, when they were in a stable clinical condition, they performed an exercise test (Incremental Shuttle Walk Test, ISWT) to measure their exercise capacity. RESULTS: Nine of 21 patients (43%) were rehospitalised within 12 month. The mean distance walked in the ISWT was 174m in patients who were hospitalised and 358m in non-hospitalised patients (P<0.001). Oxygen saturation 88% after the ISWT was found in 73% of hospitalised patients in contrast to only 22% in non-hospitalised patients (P<0.05). Activity related health status (SGRQ-activity) was higher (worse) in hospitalised patients than in non-hospitalised patients (75 vs. 50) (P<0.05). The association between walking distance and the risk of rehospitalisation was significant after adjusting for oxygen saturation and health status (hazard risk ratio 0.8 (0.67-0.97) per 10m). This study has shown that walking distance is a good and reliable predictor of rehospitalisations in moderately and severely disabled patients with COPD. PMID- 17085031 TI - PCDD/F in source-sorted waste fractions and emissions from their co-combustion with reed canary-grass. AB - The dry combustible fraction of source-sorted household waste, including material that would otherwise be recycled, was mixed with the energy crop reed canary grass (Phalaris arundinacea L.), and combusted as briquettes in 150 and 600 kW biofuel-boilers without advanced cleaning systems. The source-sorted waste was further sorted and characterized according to its material and chemical contents. The bulk of the waste's chlorine content came from the non-package plastic fraction, whereas 90-95% of summation operator PCDD/F (74-90% of WHO-TEQ) originated from the textile fraction. The sources of the dioxins in the waste fractions are discussed. The balance of dioxin levels was negative, i.e., the amounts of dioxins output in the flue gas were lower than those input in the fuel, except when there were operational disturbances in the combustion. In one of the combustion trials the total levels of dioxins in the flue-gas and ashes were also lower than the input levels. The use of additional cleaning equipment will be needed to ensure that emissions of dioxins and hydrochloric acid will be below legal limits. PMID- 17085032 TI - Studies on sildenafil citrate (Viagra) interaction with DNA using electrochemical DNA biosensor. AB - The interaction of sildenafil citrate (Viagra) with DNA was studied by using an electrochemical DNA biosensor. The binding mechanism of sildenafil citrate was elucidated by using constant current potentiometry and differential pulse voltammetry at DNA-modified glassy carbon electrode. The decrease in the guanine oxidation peak area or peak current was used as an indicator for the interaction in 0.2M acetate buffer (pH 5). The binding constant (K) values obtained were 2.01+/-0.05 x 10(5) and 1.97+/-0.01 x 10(5)M(-1) with constant current potentiometry and differential pulse voltammetry, respectively. A linear dependence of the guanine peak area or peak current was observed within the range of 1-40 microM sildenafil citrate with slope=-2.74 x 10(-4)s/microM, r=0.989 and slope=-2.78 x 10(-3)microA/microM, r=0.995 by using constant current potentiometry and differential pulse voltammetry, respectively. Additionally, binding constant values for sildenafil citrate-DNA interaction were determined for the pH range of 4-8 and in biological fluids (serum and urine) at pH 5. The influence of sodium and calcium ions was also studied to elucidate the mechanism of sildenafil citrate-DNA interaction under different solution conditions. The present study may prove to be helpful in extending our understanding of the anticancer activity of sildenafil citrate from cellular to DNA level. PMID- 17085033 TI - Electrochemical biosensing of pesticide residues based on affinity biocomposite platforms. AB - A novel and very sensitive electrochemical immunosensing strategy for the detection of atrazine based on affinity biocomposite transducers is presented. Firstly, the graphite-epoxy composite transducer was bulk-modified with different universal affinity biomolecules, such as avidin and Protein A. Two strategies for the immobilization of the anti-atrazine antibodies on both biocomposite transducers were evaluated: 'wet-affinity' and 'dry-assisted affinity' immobilization. Finally, the performance of a novel anti-atrazine immunocomposite bulk-modified with anti-atrazine antibodies was also evaluated. The better immobilization performance of the anti-atrazine antibodies was achieved by 'dry assisted affinity' immobilization on Protein A (2%) graphite-epoxy biocomposite (ProtA(2%)-GEB) as a transducer. The immunological reaction for the detection of atrazine performed on the ProtA(2%)-GEB biosensors is based on a direct competitive assay using atrazine-HRP tracer as the enzymatic label. The electrochemical detection is thus achieved through a suitable substrate and a mediator for the enzyme HRP. This novel strategy was successfully evaluated using spiked orange juice samples. The detection limit for atrazine in orange juices using the competitive electrochemical immunosensing assay was found to be 6 x 10( 3) microgL-1 (0.03 nmolL-1) thus this biosensing method accomplishes by far the LODs required for the European Community directives for potable water and food samples (0.1 microgL-1). This strategy offers great promise for rapid, simple, cost effective, and on-site biosensing of biological, food, and environmental samples. PMID- 17085034 TI - Improvement of yeast biochip sensitivity using multilayer inorganic sol-gel substrates. AB - Today, microarray fluorescence detection is still limited because a great proportion of hybrids remain undetectable. In this paper we describe sol-gel optical multilayers (stacks of low- and high-index layers) deposited on glass slides which increase the fluorescence of DNA microarrays and favour the detection of fluorescent targets. An alternative to the expensive and time consuming physical vapour deposition technology is proposed. It is a low-cost sol gel coating of glass slides, each layer being made by "dipping" (alternatively in SiO2 or TiO2 solutions), "draining and drying". After the selection of the best surface layer of the substrates, the multilayer mirrors modelled for one (Cy3) or two (Cy3 and Cy5) fluorophores are spotted with a series of Yeast probes and compared to similar microarrays on standard glass slides through hybridisation experiments. The fluorescence images of the mirrors show increased signals for all the probes. The enhancement factors determined for Cy3 and for Cy3/Cy5 mirrors (10-12 and 4-5, respectively) are consistent with the initial modelling. This allows the assessment of the basal expression levels of Yeast low-expressed genes. Moreover, these substrates show a noticeable increase in sensitivity for induction/repression ratio measurements in differential gene expression experiments. So, they could be considered as promising tools for the analysis of small biological samples. PMID- 17085035 TI - Targeting the human genome. AB - In recent years, some useful nucleic-acid-based tools including antisense oligonucleotides, aptamers, ribozymes, and small interfering RNA have been developed to alter the expression of a given gene. To date, however, these methods have proven to be generally insufficient for many applications and typically have not demonstrated high delivery efficiency or high target specificity in vivo. Emerging technologies that employ artificially designed transcription factors could offer an alternative solution, as they can recognize target DNA sequences with high specificity. In addition, these artificial proteins can be used not only as transcriptional regulators but also as genome modifiers that cleave and stimulate mutations at desired positions in the genome. These nucleotide-targeting molecules must be delivered efficiently to the target cells to promote their therapeutic activity and several delivery technologies have been developed for this purpose. PMID- 17085038 TI - The role of sleep in declarative memory consolidation: passive, permissive, active or none? AB - Those inclined to relish in scientific controversy will not be disappointed by the literature on the effects of sleep on memory. Opinions abound. Yet refinements in the experimental study of these complex processes of sleep and memory are bringing this fascinating relationship into sharper focus. A longstanding position contends that sleep passively protects memories by temporarily sheltering them from interference, thus providing precious little benefit for memory. But recent evidence is unmasking a more substantial and long lasting benefit of sleep for declarative memories. Although the precise causal mechanisms within sleep that result in memory consolidation remain elusive, recent evidence leads us to conclude that unique neurobiological processes within sleep actively enhance declarative memories. PMID- 17085039 TI - The neural origins and implications of imitation, mirror neurons and tool use. AB - Several recent studies report how laboratory-raised, non-human primates exposed to tool use can exhibit intelligent behaviors, such as imitation and reference vocal control, that are never seen in their wild counterparts. Tool-use training appears to forge a novel cortico-cortical connection that underlies this boost in capacity, which normally exists only as latent potential in lower primates. Although tool-use training is patently non-naturalistic, its marked effects on brain organization and behavior could shed light on the evolution of higher intelligence in humans. PMID- 17085040 TI - Neuromodulation of central pattern generators in invertebrates and vertebrates. AB - Central pattern generators are subject to extensive modulation that generates flexibility in the rhythmic outputs of these neural networks. The effects of neuromodulators interact with one another, and modulatory neurons are themselves often subject to modulation, enabling both higher order control and indirect interactions among central pattern generators. In addition, modulators often directly mediate the interactions between functionally related central pattern generators. In systems such as the vertebrate respiratory central pattern generator, multiple pacemaker types interact to produce rhythmic output. Modulators can then alter the relative contributions of the different pacemakers, leading to substantial changes in motor output and hence to different behaviors. Surprisingly, substantial changes in some aspects of the circuitry of a central pattern generator, such as a several-fold increase in synaptic strength, can sometimes have little effect on the output of the CPG, whereas other changes have profound effects. PMID- 17085042 TI - Viral enzymes. AB - Viral genomes show unequalled diversity, ranging from single-stranded DNA to double-stranded RNA. Moreover, viruses can quickly adapt to the host's immune response and drug treatment. Although they tend to make optimal use of the host cell's reservoir of proteins, viruses need to carry some enzymatic functions with them, as they may not be available or accessible in the infected cell. Recently, progress has been made in our structural understanding of viral enzymes involved in all stages of the viral life cycle, which includes entry, hijack, replication and exit stages. PMID- 17085043 TI - Protein tyrosine kinase-substrate interactions. AB - Protein tyrosine kinases (PTKs) are enzymes that catalyze the phosphorylation of tyrosyl residues. They are important in physiological and pathophysiological processes. Protein substrates of PTKs are often difficult to discern, but recently reported methods have helped to identify targets and characterize their structural interactions with kinases. A mechanism-based bisubstrate analog strategy has given X-ray crystallographic insights into how several topical PTKs, including the insulin receptor, Abl and epidermal growth factor receptor, interact with tyrosine-containing peptide substrates. These PTK co-crystal structures reveal both conserved and specialized features of recognition that probably contribute to substrate selection and the individual functions of these key enzymes. PMID- 17085044 TI - Networks for the allosteric control of protein kinases. AB - The allosteric regulation of protein kinases serves as an efficient strategy for molecular communication, event coupling and interconversion between catalytic states. Recent co-crystal structures have revealed novel ways in which kinases control activity and substrate specificity following phosphorylation, dimerization, or binding to regulatory proteins, substrates and scaffolds. In addition, hydrogen exchange coupled with mass spectrometry is emerging as a complementary strategy to probe the solution behavior of kinases; recent results have shown that allosteric regulation may involve transitions in protein motions as well as structural rearrangements. PMID- 17085045 TI - Guidelines for the adjuvant treatment of postmenopausal women with endocrine responsive breast cancer: past, present and future recommendations. AB - Treatment guidelines are useful tools that enable physicians to integrate the latest clinical research into their practices. The large volume of rapidly evolving clinical data in breast cancer has been summarised and incorporated into treatment recommendations by well-known and reliable institutions, including the National Comprehensive Cancer Network, the American Society for Clinical Oncology, the European Society for Medical Oncology and the St. Gallen International Consensus Panel. Adjuvant therapy is a key component of breast cancer treatment, and many of the current consensus guidelines now recognise the important role of the aromatase inhibitors as an alternative to or in sequence after tamoxifen, hitherto the standard adjuvant treatment of choice for receptor positive women. Data from ongoing trials such as the Breast International Group 1 98 trial and those still in the accrual phase will be forthcoming and will likely result in a further refinement of treatment recommendations over the course of the next few years. Despite the availability of such guidelines, however, there is evidence that adherence to and implementation of treatment recommendations is less than optimal. Further research is needed to determine more effective means of disseminating those clinical recommendations that can have a significant impact on treatment strategies and ultimately improve outcomes in breast cancer. PMID- 17085046 TI - Functional genome analysis indicates loss of 17beta-hydroxysteroid dehydrogenase type 2 enzyme in the zebrafish. AB - Among the family of 17beta-hydroxysteroid dehydrogenases, the type 2 (17beta-HSD 2) is the main enzyme responsible for inactivation of estrogens and androgens, catalyzing the oxidation of the C17 hydroxyl group. 17beta-HSD 2 has been studied only in mammals, its occurrence and function in other vertebrates hardly known. We investigated the presence of homologs in non-mammalian species and found sequences of 17beta-HSD 2 and its closest homolog 11beta-HSD 2 in zebrafish (Danio rerio), Takifugu rubripes, Tetraodon nigroviridis, Xenopus tropicalis and chicken databases. Furthermore, we cloned zebrafish 17beta-HSD 2 from ovarian tissue and found high expression also in the testis of adult fish and throughout embryogenesis. The enzyme, though, is inactive likely due to a non-sense N terminal region including a dysfunctional cofactor binding motif. Replacement of the affected part by the corresponding human 17beta-HSD 2 sequence fully restored enzymatic activity. Comparison of all retrieved 17beta-HSD 2 sequences indicates that this functional loss may have occurred only in zebrafish, where steroid inactivation at position C17 seems to pursue without the protein studied. The closely related 11beta-HSD 2 is unlikely to substitute for 17beta-HSD 2 since in our hands it did not catalyze the respective oxidation of testosterone or estradiol. PMID- 17085047 TI - Emerging principles for the development of resistance to antihormonal therapy: implications for the clinical utility of fulvestrant. AB - We seek to evaluate the clinical consequences of resistance to antihormonal therapy by studying analogous animal xenograft models. Two approaches were taken: (1) MCF-7 tumors were serially transplanted into selective estrogen receptor modulator (SERM)-treated immunocompromised mice to mimic 5 years of SERM treatment. The studies in vivo were designed to replicate the development of acquired resistance to SERMs over years of clinical exposure. (2) MCF-7 cells were cultured long-term under SERM-treated or estrogen withdrawn conditions (to mimic aromatase inhibitors), and then injected into mice to generate endocrine resistant xenografts. These tumor models have allowed us to define Phase I and Phase II antihormonal resistance according to their responses to E(2) and fulvestrant. Phase I SERM-resistant tumors were growth stimulated in response to estradiol (E(2)), but paradoxically, Phase II SERM and estrogen withdrawn resistant tumors were growth inhibited by E(2). Fulvestrant did not support growth of Phases I and II SERM-resistant tumors, but did allow growth of Phase II estrogen withdrawn-resistant tumors. Importantly, fulvestrant plus E(2) in Phase II antihormone-resistant tumors reversed the E(2)-induced inhibition and instead resulted in growth stimulation. These data have important clinical implications. Based on these and prior laboratory findings, we propose a clinical strategy for optimal third-line therapy: patients who have responded to and then failed at least two antihormonal treatments may respond favorably to short-term low-dose estrogen due to E(2)-induced apoptosis, followed by treatment with fulvestrant plus an aromatase inhibitor to maintain low tumor burden and avoid a negative interaction between physiologic E(2) and fulvestrant. PMID- 17085048 TI - Nitrogen-containing flavonoids as CDK1/Cyclin B inhibitors: design, synthesis, and biological evaluation. AB - A novel series of nitrogen-containing flavonoids 5a-l, 6a,b, and 7a,b were designed and synthesized as cyclin-dependent kinases (CDKs) inhibitors. The representative compounds 5a, 5b, 5e, and 5g showed potent CDK1/Cyclin B inhibitory activities. All compounds displayed a significant growth inhibitory action in vitro against Bel-7402, PC-3, ECA-109, A-549, HL-60, and MCF-7 cancer cell lines. Flow cytometry analysis showed that 5b induced apoptosis in PC-3 cells. PMID- 17085049 TI - Phospholipase C epsilon: linking second messengers and small GTPases. AB - Although several observations over the years suggested a link between the Ras superfamily of GTPases and second messengers generated by the isoforms of phospholipase C, such links had not been substantiated at the molecular level until recently. In particular, identification of a novel phospholipase C isoform, PLC epsilon, which also incorporates domains for guanine nucleotide exchange and Ras binding, have prompted an interest in the interplay between small GTPases and phospholipase C and possible significance of these interconnectivities. Research that followed suggests that activation of each of the major classes of phospholipase C by small GTPases could have a different mechanism and different function, and also that phospholipase C enzymes in turn control Ras GTPases through regulatory proteins that respond directly to second messengers. PMID- 17085050 TI - ErbB receptors: new insights on mechanisms and biology. AB - The ErbB family of four receptor tyrosine kinases occupies a central role in a wide variety of biological processes from neuronal development to breast cancer. New information continues to expand their biologic significance and to unravel the molecular mechanisms that underlie the signaling capacity of these receptors. Here, we review several aspects of ErbB receptor physiology for which new and significant information is available. These include ligand-dependent receptor dimerization and kinase activation, which is a prerequisite for all subsequent growth factor-dependent cell responses. We also address novel roles of receptor fragments in signaling, trafficking to intracellular sites, such as the nucleus, and ErbB roles in non-cancer disease processes, including schizophrenia, chronic renal disease, hypertension, and the cellular entry of infectious pathogens. PMID- 17085051 TI - Preparation of secolycorines against acetylcholinesterase. AB - 5,6-Secolycorines possessing a 5,6-dihydrophenanthridine skeleton were facilely prepared from lycorine through chemical transformations, mainly including N alkylation, Hofmann degradation type reaction, reductive cleavage of trichloroethylcarbonyl moiety, and hydrogenation. Several secolycorine derivatives showed potent inhibitory activity against acetylcholinesterase with the IC(50) value at micromolar range and are more potent than galanthamine. PMID- 17085052 TI - Molecular probes of DNA bulges: functional assay and spectroscopic analysis. AB - Bulged structures in DNA and RNA have been linked to biomolecular processes involved in numerous diseases, thus probes with affinity for these nucleic acid targets would be of considerable utility to chemical biologists. Herein, we report guided chemical synthesis of small molecules capable of binding to DNA bulges by virtue of their unique (spirocyclic) geometry. The agents, modeled on a natural product congener, show pronounced selectivity for specific bulged motifs and are able to enhance slipped DNA synthesis, a hallmark functional assay of bulge binding. Significantly, bulge-agent complexes demonstrate characteristic fluorescent signatures depending on bulge and flanking sequence in the oligo. It is anticipated that these signature patterns can be harnessed as molecular probes of bulged hotspots in DNA and RNA. PMID- 17085053 TI - Enantioselective synthesis and antiviral activity of purine and pyrimidine cyclopentenyl C-nucleosides. AB - The enantiomerically pure carbocyclic purine and pyrimidine C-nucleosides 1-4 were synthesized via the key intermediate, 2,3-(isopropylidenedioxy)-4 (trityloxymethyl)-4-cyclopenten-1-ol (5), which was prepared from d-ribose in eight steps. Synthesized compounds were evaluated as potential antiviral agents against HIV, SARSCoV, Punta Toro, West Nile, and Cowpox viruses. However, only 9 deazaneplanocin A (1) exhibited moderate anti-HIV activity. PMID- 17085054 TI - Synthesis and antitumor activities of a series of novel quinoxalinhydrazides. AB - Recently, we discovered a novel class of anticancer compounds with remarkable potency in a panel of cancer cell lines. A prototype compound, SC144, showed significant in vivo efficacy in mice xenograft models of human breast cancer cells. Herein, we report on a new synthetic route to SC144 and the synthesis of several of its analogues in order to understand required features for activity. A one-step coupling of 7-fluoro-4-chloropyrrolo[1,2-a]quinoxaline with pyrazin-2 carbohydrazide improved the yield significantly. Although several of the analogues showed significant activities, modification of the heteroacyl moiety had a dramatic effect on potency. PMID- 17085055 TI - Comparison of the TL responses of two different preparations of LiF:Mg,Cu,P irradiated by photons of various energies. AB - The aim of this work is to present the results concerning the photon irradiation of a new phosphor, the LiF:Mg,Cu,P+PTFE, produced at the Instituto Nacional de Investigaciones Nucleares (ININ-Mexico). The photon irradiations were performed using X-rays of 16, 24, 34.5, 42, 100 and 145keV, and gamma rays from (137)Cs (662keV) and from (60)Co (1 252keV). The results obtained are normalized to the (60)Co response. The experimental data are then compared to those obtained using the commercial dosimeters TLD-100 and GR-200A. PMID- 17085056 TI - Anal carcinoma of the elderly treated with radiotherapy alone or with concomitant radio-chemotherapy. AB - PURPOSE: To analyse the results achieved with radio-chemotherapy (RTCT) or radiotherapy alone (RT) in elderly patients (pts) affected with squamous cell anal cancer. METHODS AND MATERIALS: From 1990 to 2002, 62 pts aged > or =70 years were treated with RT (14) or RTCT (48). There were 9 stage I, 29 stage II, 11 stage IIIa and 13 stage IIIb. MMC+5FU was given concomitantly with RT in an early period, later replaced by Cddp+5FU. In the RTCT group, 36Gy were delivered to pelvic+inguinal lymph nodes, with a tumor boost (18Gy). RESULTS: Stage II fared significantly better than stage III in terms of locoregional control (LRC) but not overall survival (OS). Pts treated with RTCT had improved LRC, but not OS. LRC was 81% at 3 and 5 years for the RTCT group; the RT group had a LRC of 61% at 3 years. There were more locoregional relapses in the MMC group (29%) versus the Cddp group (19%) and in pts treated with a split (32%) versus no split (19%). No G3 acute toxicity was observed in the RT group; in the RTCT group 15 pts (31%) developed a G3+ acute toxicity. G3+ late damage occurred in 2 pts in the RT only group and in 3 pts in the RTCT group. CONCLUSIONS: Elderly people considered fit for RTCT should undergo the same schedules used for younger people. MMC or Cddp+5FU are feasible in the elderly, even without a planned split. PMID- 17085057 TI - Broad-spectrum L-amino acid sensing by class 3 G-protein-coupled receptors. AB - The sensing of nutrients is essential to the control of growth and metabolism. Although the sensing mechanisms responsible for the detection and coordination of metabolic responses to some nutrients, most notably glucose, are well understood, the molecular basis of amino acid sensing by cells and tissues is only now emerging. In this article, we consider evidence that some members of G-protein coupled receptor class 3 are broad-spectrum amino acid sensors that couple changes in extracellular amino acid levels to the activation of intracellular signaling pathways. In particular, we consider both the molecular basis of specific and broad-spectrum amino acid sensing by different members of class 3 and the physiological significance of broad spectrum amino acid sensing by the extracellular calcium-sensing receptor, heterodimeric taste receptors and the recently "deorphanized" receptor GPRC6A and its goldfish homolog, the 5.24 chemoreceptor. PMID- 17085058 TI - Wide range of susceptibility to rhabdoviruses in homozygous clones of rainbow trout. AB - Inbred lines differentially susceptible to diseases are a powerful tool to get insights into the mechanisms of genetic resistance to pathogens. In fish, chromosome manipulation techniques allow a quick production of such homozygous lines. Using gynogenesis, we produced nine homozygous clones of rainbow trout from a domestic population (INRA Sy strain). We examined the variability between clones for resistance to two rhabdoviruses, the viral haemorrhagic septicaemia virus (VHSV) and the infectious haematopoietic necrosis virus (IHNV). Intraperitoneal injections and waterborne infections were performed in parallel for both viruses. No survival was recorded after intraperitoneal injection of VHSV or IHNV, indicating that fish from all clones were fully susceptible to both viruses by this route of infection. In contrast, the different clones showed a wide range of survival frequency after waterborne infection. The resistance levels to VHSV ranged from 0 to 99% and resistance was not abrogated when resistant and sensitive animals were mixed and subjected to waterborne infection. VHSV was recovered from 10% of resistant fish after waterborne infection, confirming that virus replication was possible in this context but effective only in a low proportion of the population. The different clones also exhibited a wide range of survival (0-68%) after a waterborne infection with IHNV. Although VHSV resistant clones were not fully resistant to IHNV, the susceptibility to IHNV and VHSV tended to be correlated, suggesting that non-specific mechanisms common to both viruses were involved. PMID- 17085059 TI - Is energy expenditure taken into account in human sub-maximal jumping?--A simulation study. AB - This paper presents a simulation study that was conducted to investigate whether the stereotyped motion pattern observed in human sub-maximal jumping can be interpreted from the perspective of energy expenditure. Human sub-maximal vertical countermovement jumps were compared to jumps simulated with a forward dynamic musculo-skeletal model. This model consisted of four interconnected rigid segments, actuated by six Hill-type muscle actuators. The only independent input of the model was the stimulation of muscles as a function of time. This input was optimized using an objective function, in which targeting a specific sub-maximal height value was combined with minimizing the amount of muscle work produced. The characteristic changes in motion pattern observed in humans jumping to different target heights were reproduced by the model. As the target height was lowered, two major changes occurred in the motion pattern. First, the countermovement amplitude was reduced; this helped to save energy because of reduced dissipation and regeneration of energy in the contractile elements. Second, the contribution of rotation of the heavy proximal segments of the lower limbs to the vertical velocity of the centre of gravity at take-off was less; this helped to save energy because of reduced ineffective rotational energies at take-off. The simulations also revealed that, with the observed movement adaptations, muscle work was reduced through improved relative use of the muscle's elastic properties in sub-maximal jumping. According to the results of the simulations, the stereotyped motion pattern observed in sub-maximal jumping is consistent with the idea that in sub-maximal jumping, subjects are trying to achieve the targeted jump height with minimal energy expenditure. PMID- 17085060 TI - vasa-related genes and their expression in stem cells of colonial parasitic rhizocephalan barnacle Polyascus polygenea (Arthropoda: Crustacea: Cirripedia: Rhizocephala). AB - vasa (vas)-related genes are members of the DEAD-box protein family and are expressed in the germ cells of many Metazoa. We cloned vasa-related genes (PpVLG, CpVLG) and other DEAD-box family related genes (PpDRH1, PpDRH2, CpDRH, AtDRHr) from the colonial parasitic rhizocephalan barnacle Polyascus polygenea, the non colonial Clistosaccus paguri (Crustacea: Cirripedia: Rhizocephala), and the parasitic isopodan Athelgis takanoshimensis (Crustacea: Isopoda). The colonial Polyascus polygenea, a parasite of the coastal crabs Hemigrapsus sanguineus and Hemigrapsus longitarsis was used as a model object for further detailed investigations. Phylogenetic analysis suggested that PpVLG and CpVLG are closely related to vasa-like genes of other Arthropoda. The rest of the studied genes form their own separate branch on the phylogenetic tree and have a common ancestry with the p68 and PL10 subfamilies. We suppose this group may be a new subfamily of the DEAD-box RNA helicases that is specific for parasitic Crustacea. We found PpVLG and PpDRH1 expression products in stem cells from stolons and buds of internae, during asexual reproduction of colonial P. polygenea, and in germ cells from sexually reproducing externae, including male spermatogenic cells and female oogenic cells. PMID- 17085061 TI - Differential expression of phospholipases D1 and D2 in mouse tissues. AB - The differential expression of phospholipase D (PLD) isozymes, which include PLD1 and PLD2, was examined in various murine tissues, including the cerebrum, cerebellum, heart, lung, liver, spleen, stomach, pancreas, ileum, colon, adrenal gland, kidneys, testes, ovaries, and uterus. In Western blot analysis, only PLD1 was detected in the heart and ovary, while only PLD2 was detected in the pancreas and ileum. Both PLD1 and PLD2 were strongly expressed in the cerebrum, cerebellum, and lung, and both were also expressed in the liver, spleen, stomach, colon, kidney, testes, and uterus. Immunohistochemistry showed intense PLD immunostaining in the cerebrum, cerebellum, lungs, intestines, and testis, and weak PLD immunostaining in the liver, kidneys, spleen, and heart. These findings suggest that PLD1 and PLD2 are differentially expressed in the various organs of mice, and that each PLD isozyme plays a distinct role in each organ. PMID- 17085062 TI - Inherited metabolic disease in companion animals: searching for nature's mistakes. AB - Inborn errors of metabolism are caused by genetic defects in intermediary metabolic pathways. Although long considered to be the domain of human paediatric medicine, they are also recognised with increasing frequency in companion animals. The diagnosis of diseased animals can be achieved by searching for abnormal metabolites in body fluids, although such screening programmes have, until now, not been widely available to the small animal clinician. A comprehensive battery of analytical tools exists for screening for inborn metabolic diseases in humans which can be applied to animals and serve not only for the diagnosis of affected patients but also to detect asymptomatic carriers and further our understanding of metabolic pathways in dogs and cats. Moreover, naturally occurring animal models of inherited metabolic diseases provide a unique opportunity to study the biochemical and molecular pathogenesis of these disorders and to investigate possible therapeutic options. PMID- 17085063 TI - A benign congenital myopathy in an inbred Samaritan family. PMID- 17085064 TI - [Is there still a place for extemporaneous exam in breast cancer?]]. PMID- 17085066 TI - The use of the Reichert ocular response analyser to establish the relationship between ocular hysteresis, corneal resistance factor and central corneal thickness in normal eyes. AB - PURPOSE: The aim of this study was to measure ocular hysteresis and corneal resistance factor (CRF), novel methods of analysing ocular rigidity/elasticity and to determine the relationship between central corneal thickness (CCT), hysteresis and CRF in normal subjects. DESIGN: Prospective, cross-sectional, clinical trial. PARTICIPANTS: The study included 207 normal eyes. METHODS: Hysteresis and CRF were measured by the ocular response analyser. The CCT was measured using a hand held ultrasonic pachymeter. MAIN OUTCOME MEASURES: Ocular hysteresis and CRF in normal patients and their relationship with CCT. RESULTS: The mean hysteresis was 10.7+/-2.0 mmHg standard deviation (S.D.) (range 6.1-17.6 mmHg); the mean CRF was 10.3+/-2.0 (range 5.7-17.1 mmHg). The mean CCT was 545.0+/-36.4 microm (471-650 microm). The relationship between hysteresis and CCT; CRF and CCT; CRF and hysteresis were significant (p<0.0001). CONCLUSION: This study demonstrated that corneal hysteresis increased with increasing CCT, however, the correlation was moderate. It would appear that CCT, hysteresis and CRF may measure different biomechanical aspects of ocular rigidity and are likely to be useful additional measurement to CCT in the assessment of ocular rigidity when measuring intraocular pressure (IOP). This may be of particular importance when trying to correct IOP measurements for increased or decreased ocular rigidity. PMID- 17085067 TI - Polynuclear transition metal complexes with thiocarbohydrazide and dithiocarbamates. AB - Sn(tch)2{MCl2}2 was prepared from the precursor Sn(tch)2 and MCl2. It was subsequently allowed to react with diethyldithiocarbamate which yielded the trinuclear complexes of the type Sn(tch)2{M2(dtc)4}, where tch=thiocarbohydrazide, M=Mn(II), Fe(II), Co(II), Ni(II), Cu(II) and dtc=diethyldithiocarbamate. They were characterized on the basis of microanalytical, thermal (TGA/DSC), spectral (IR, UV-vis, EPR, (1)H NMR) studies, conductivity measurement and magnetic moment data. On the basis of spectral data a tetrahedral geometry has been proposed for the halide complexes, Sn(tch)2{MCl2}2 except for Cu(II) which exhibits a square planar coordination although the transition metal ion in Sn(tch)2{M2(dtc)4} achieves an octahedral geometry where the dithiocarbamato moiety acts as a symmetrical bidentate ligand. The bidentate nature has been established by the appearance of a sharp single nu(C-S) around 1000 cm(-1). A downfield shift observed in NH(a) and NH(b) protons on moving from Sn(tch)2 to Sn(tch)2{MCl2}2 is due to the drift of electrons toward metal atoms. A two-step pyrolysis has been observed in the Sn(tch)2{MCl2}2 complexes while their dithiocarbamato derivatives exhibit a three-stage degradation pattern. Finally, the in vitro antibacterial activity of Sn(tch)2{M2(dtc)4} and the mononuclear Sn(tch)2 has been carried out on bacterial strains Escherichia coli and Salmonella typhi. The compounds were found to be active against the test organisms. The activity of the complexes is enhanced with increasing concentration. The maximum activity in both the strains was achieved by cobalt(II) dithiocarbamate complex. Minimum activity was found for Sn(tch)2 which generally increases with the introduction of transition metal ion in the complex. PMID- 17085068 TI - Mono-, di-, and tri-tert-butyl ethers of glycerol. A molecular spectroscopic study. AB - MS, NMR, IR and Raman molecular spectroscopy techniques were applied to characterize 3-tert-butoxy-propane-1,2-diol, 1,3-di-tert-butoxy-propan-2-ol, and 1,2,3-tri-tert-butoxy-propane. These ethers are the main products of glycerol etherification reaction and are excellent oxygen additives for diesel fuel. Computational DFT/ B3LYP/6-31G** studies were performed to support and rationalize both vibrational spectroscopy analysis and the isomer ratio. PMID- 17085069 TI - Unique patient concept: a key choice for European epidemiology. AB - The French Ministry of Health is setting up the personal medical record (PMR or DMP for Dossier Medical Personnel in French). This innovating tool is highly expected and will be extremely useful for the therapeutic follow-up as well as for epidemiological studies on which public health policies are based. Therefore the currently planned identifying process should prevent any epidemiological use of these data. Numerous scientific organisations have alerted government powers about the threat that this impairment represents, and they wish to promote some secure procedures that exist, which have already proved their efficiency at the national and international level. PMID- 17085070 TI - Human metapneumovirus infections among children with acute respiratory infections seen in a large referral hospital in India. AB - BACKGROUND: Acute respiratory infections (ARI) are the leading cause of morbidity and mortality among children <5 years of age in developing countries. Human metapneumovirus (hMPV), a newly described respiratory pathogen, has been identified as an important cause of ARI in young children. OBJECTIVES: The objective was to describe the prevalence of hMPV in children who presented with ARI to a large referral hospital in Delhi, India and to genotype circulating strains on the basis of F gene nucleotide sequence analysis. STUDY DESIGN: We analyzed 97 samples from children <5 years of age with ARI seen at the All India Institute of Medical Sciences from June 2004 to March 2005. RT-PCR was performed for the N and F genes and partial F gene nucleotide sequences were used to characterize the viruses. RESULTS: hMPV was identified in 12% of children with ARI, including 13% of the children hospitalized with ARI. Most virus identification occurred in the winter. The Indian strains clustered in the A2 genetic sublineage. CONCLUSIONS: This report establishes hMPV as an important cause of ARI in children in India. PMID- 17085071 TI - Analysis of biomolecular data: a post-genomic reflection. PMID- 17085072 TI - Effects of separate and combined exposure to the pesticides methamidophos and chlorothalonil on the development of suckling rats. AB - It has been suggested that, in some cases, exposure to environmental contaminants affects children more profoundly than adults. It is important to evaluate adverse health outcomes in children, a population susceptible to toxic chemicals and mixtures. We examined the effects that maternal exposure to two pesticides had on maturational aspects of offspring development during the nursing period. Nursing female rats were exposed to 1-4mg/kg of intraperitoneal methamidophos, 200 800mg/kg of chlorothalonil, or both. The higher doses of methamidophos affected pup viability by day 21 of life. Both pesticides, alone or together, affected body weight gain of dams and offspring. Developmental milestones evaluated in the pups were incisor eruption, ear unfolding, eye opening and testis descent. Although no clear dose-response relationship was established between these milestones and exposure to methamidophos or chlorothalonil, incisor eruption was accelerated in many groups, and the majority of rat offspring exposed to methamidophos presented later ear unfolding and eye opening than did the control group offspring. Sexual maturation (testis descent) was significantly delayed in some groups. For dams and pups alike, simultaneous exposure to both pesticides was not found to have a greater toxic effect than that resulting from exposure to only one of the two. Taken together, these results demonstrate exposure-related influences on several developmental measures. Detection of more subtle effects may be improved through the use of the developmental temporal response protocols utilized in this study. PMID- 17085073 TI - p11 (S100A10)--an inducible adaptor protein that modulates neuronal functions. AB - p11 (S100A10) is a member of the S100 protein family and forms a heterotetrameric complex with annexin 2. p11 has also been found to interact with a diverse set of proteins that includes several ion channels and the serotonin 5-HT1B receptor. Several factors such as dexamethasone, growth factors, nitric oxide and antidepressant therapies regulate the expression of p11. Furthermore, studies using mutant mouse models, RNA interference and antisense constructs have implicated p11 in several biological processes; in particular, there is evidence that p11 is involved in the pathophysiology underlying nociception and depression like states. PMID- 17085074 TI - ERK2: a logical AND gate critical for drug-induced plasticity? AB - Drug addiction results in part from the distortion of dopamine-controlled plasticity, and extracellular signal-regulated kinase (ERK) plays an important role in the underlying molecular mechanisms of this process. ERK is activated by drugs of abuse in a subset of neurons in reward-related brain regions. This activation, necessary for the expression of immediate early genes, depends upon dopamine D1 and glutamate receptors. Blockade of ERK activation prevents long lasting behavioral changes, including psychomotor sensitization and conditioned place preference. It also interferes with drug craving and drug-associated memory reconsolidation. By contrast, ERK1 mutation enhances the effects of morphine and cocaine. We suggest that the ERK2 pathway acts as a logical AND gate, permissive for plasticity, in neurons on which dopamine-mediated reward signals and glutamate-mediated contextual information converge. PMID- 17085075 TI - Role of anti-nucleosome antibody in the diagnosis of systemic lupus erythematosus. AB - OBJECTIVE: To investigate the role of anti-nucleosome antibodies (AnuA) in systemic lupus erythematosus (SLE). METHODS: IgG anti-nucleosome antibodies were detected by enzyme-linked immunosorbent assay (ELISA) in the sera of 233 SLE and 220 other rheumatic diseases' patients as well as 31 health controls. The patients were also evaluated for clinical and biological parameters. RESULTS: (1) Out of 233 SLE patients, 144 (61.8%) were seropositive for AnuA, which was significantly higher than that of patients with other rheumatic diseases [2.7% (6/220), P < 0.001]; the sensitivities and specificities of AnuA in SLE were 61.8% and 97.6%, respectively. (2) The positive rated of AnuA in SLE lacking of anti-DNP, anti-cmDNA, anti-Sm and anti-dsDNA antibodies were 57.1%, 55.9%, 62.4% and 51.2%, respectively. (3) The frequency of the fever, skin rash, and arthralgia were significantly higher in 144 positive AnuA SLE than those in AnuA negative SLE (P < 0.05). The frequency of leukopenia, elevations of ESR and CRP, lower C3/C4 levels and proteinuria in AnuA positive groups was significantly higher than that of AnuA negative groups (P < 0.05). (4) Level of AnuA was strongly correlated with the SLEDAI scores (r = 0.385, P < 0.001). Patients with active SLE showed significantly higher positive rate of AnuA (66.1%) than those with inactive disease (45.7%) (chi2 = 6.568, P = 0.010). CONCLUSIONS: AnuA is one of the most valuable markers in the diagnosis of SLE lacking of anti-dsDNA, anti Sm, anti-DNP and anti-cmDNA antibodies. The level of AnuA is associated with the disease activity of SLE. PMID- 17085076 TI - Quality management systems in ART: are they really needed? An Australian clinic's experience. AB - As assisted reproductive technology (ART) expanded globally, several countries introduced prescribed requirements for treatment and monitoring of outcomes, as well as a licensing or accreditation requirement. While it is common for ART laboratories to be required to have an effective quality control system, the remainder of the clinic is often under less stringent regulation. Furthermore, when treatment conditions are prescribed, the standards tend to be conservative and clinics may choose to establish their own standards. Total quality management systems are now being used by an increasing number of ART clinics. In Australia and New Zealand, it is now a requirement to have a quality management system in order to be accredited and to help meet customer demand for improved delivery of ART services in these two countries. PMID- 17085077 TI - Donor gametes: anonymous or identified? AB - The practice of gamete donation has, until recently, been shrouded in secrecy. The stigma associated with infertility and, in particular, donor insemination has been the main factor contributing to this secrecy. Over the last 20 years, this secrecy and the anonymity of the gamete donors has been challenged. In the first instance, the challenge came from governments in some countries legislating to abolish donor anonymity. Counsellors, social workers and psychologists advocating for the interests and needs of children and their families, as well as parents who did not wish to keep gamete donation secret from their children, were also instrumental in the change of policies and practice. Those offspring who know that they were conceived as a result of gamete donation are also calling for an end to the secrecy. This chapter reviews the changes that have occurred and which are still occurring, and reviews the research associated with these changes. PMID- 17085078 TI - Zebrafish Angiotensin II Receptor-like 1a (agtrl1a) is expressed in migrating hypoblast, vasculature, and in multiple embryonic epithelia. AB - The human gene AGTRL1 is an angiotensin II receptor-like gene expressed in vasculature, which acts as the receptor for the small peptide APELIN, and a co receptor for Human Immunodeficiency Virus. Mammalian AGTRL1 has been shown to modulate cardiac contractility, venous and arterial dilation, and endothelial cell migration in vitro, but no role in the development of the vasculature, or other tissues, has been described. We report the identification and expression of the zebrafish ortholog of the human gene AGTRL1. Zebrafish agtrl1a is first expressed before epiboly in dorsal precursors. During epiboly it is expressed in the enveloping layer, yolk syncytial layer and migrating mesendoderm. During segmentation stages, expression is observed in epithelial structures such as adaxial cells, border cells of the newly formed somites, developing lens, otic vesicles and venous vasculature. PMID- 17085079 TI - Novel transcript nort is a downstream target gene of the Notch signaling pathway in zebrafish. AB - The Notch signaling pathway plays important roles in the regulation of diverse developmental processes. Although many Notch-signal target genes with different specificities have been identified, their regulation and functions are not fully understood. Here, we conducted a microarray screen to search for novel downstream target genes of the Notch pathway in zebrafish. From the screen, we isolated nort (Notch-regulated transcript) as a transcript whose expression was reduced by the inhibition of Notch signaling. The expression level of nort increased when Notch signaling was activated. nort was expressed in hypoblast cells and the developing nervous system. We found its expression pattern to be similar to that of her4, but it showed some differences, at least in the anterior and posterior neural plate at the 3-somite stage. The nort transcript did not contain any long open reading frame (ORF) of more than 300 nt, and its ORF-encoded sequence showed no significant homology with the proteins in databases. However, nort has one SPS (suppressor of hairless paired binding site) in its 5'-flanking region. These data suggest that nort is a putative noncoding RNA regulated by Notch signaling. PMID- 17085080 TI - Three tandem HRDC domains have synergistic effect on the RecQ functions in Deinococcus radiodurans. AB - The RecQ family of DNA helicases performs essential functions in the maintenance of genomic stability in all organisms. In Deinococcus radiodurans, DR1289 is a special member of RecQ family with unique arrangement of three tandem HRDC domains in the C-terminus. A dr1289 mutant is hypersensitive to gamma irradiation, UV, H2O2 and mitomycin C. By complementing the dr1289 mutant with various domains of Dr1289 in vivo, we have determined that the helicase and all three HRDC domains are indispensable for complete DNA damage resistance. Using a continuous fluorescent dye-displacement assay, we investigated the optimal conditions for Dr1289 unwinding function at various concentrations of ATP and metal ions to show that the helicase activity is comparable to what observed in Escherichia coli RecQ. We also found that the helicase domain is necessary for the unwinding and ATPase activity and that the three tandem HRDC domains increase the efficiency of these activities. Based on these data, we propose that the C terminus of Dr1289 has evolved in D. radiodurans to confront the types and amounts of DNA damage. PMID- 17085081 TI - Studies on neurosteroids XIX. Development and validation of liquid chromatography tandem mass spectrometric method for determination of 5alpha-reduced pregnane type neurosteroids in rat brain and serum. AB - A sensitive liquid chromatography-electrospray ionization-tandem mass spectrometric (LC-ESI-MS-MS) method for the simultaneous determination of 5alpha reduced pregnan-type neurosteroids, allopregnanolone (AP), epiallopregnanolone and 5alpha-dihydroprogesterone, in rat brain and serum has been developed and validated. The brain and serum steroids were extracted with methanol-acetic acid, purified using a Strata-X cartridge, derivatized with the permanently charged reagent, 2-hydrazino-1-methylpyridine (HMP), and subjected to LC-positive ESI-MS MS. The limits of quantitation (LOQ) for brain (0.25 ng/g tissue) and serum (0.25 ng/ml) assays using the derivatization-ESI-MS-MS method are 60-150-fold lower than the LOQs for their atmospheric pressure chemical ionization-MS method without derivatization. [17Alpha,21,21,21-2H4]-AP was used as an internal standard. This method allowed the reproducible and accurate quantification of the brain or serum neurosteroids using a 20 mg or 20 microl sample, respectively. That is, the intra- and inter-assay coefficients of variation were below 8.2 and 6.0%, respectively, and the % accuracy values were 98.5-103.0% for all the steroids in both the brain and serum. The application of the developed method to the analysis of changes in the brain and serum neurosteroid levels by immobilization stress and ethanol administration is also presented. PMID- 17085082 TI - Response surface methodology for the evaluation of glucose-6-phosphate dehydrogenase enrichment process by soybean lecithin reversed micelles. AB - Glucose-6-phosphate dehydrogenase (G6PD) present in Saccahromyces cerevisiae is an enzyme of the pentose pathway. An effective enrichment of this intracellular enzyme can be achieved with the reversed micellar methodology. In this work, this methodology was employed with soybean lecithin, a biocompatible surfactant. A factorial design was used to evaluate the influence of pH (A) and extraction runs (B) on the G6PD purification factor. After statistical analysis and process optimization, a mathematical model representing G6PD enrichment was obtained: Y=4.89-0.83A+0.092B+0.27AB-1.37B2 with an enzyme purification factor of about 5.2. PMID- 17085083 TI - A developed determination of midazolam and 1'-hydroxymidazolam in plasma by liquid chromatography-mass spectrometry: application of human pharmacokinetic study for measurement of CYP3A activity. AB - This paper describes sensitive and reliable determination of midazolam (MDZ) and its major metabolite 1'-hydroxymidazolam (1-OHMDZ) in human plasma by liquid chromatography-mass spectrometry (LC-MS) with a sonic spray ionization (SSI) interface. MDZ, 1-OHMDZ and diazepam as an internal standard were extracted from 1ml of alkalinized plasma using n-hexane-chloroform (70:30, v/v). The extract was injected into an analytical column (YMC-Pak Pro C(18), 50mmx2.0mmi.d.). The mobile phase for separation consisted of 10mM ammonium acetate and methanol (50:50, v/v) and was delivered at a flow-rate of 0.2ml/min. The drift voltage was 100V. The sampling aperture was heated at 120 degrees C and the shield temperature was 260 degrees C. The total time for chromatographic separation was less than 16min. The validated concentration ranges of this method were 0.25 50ng/ml for both MDZ and 1-OHMDZ. Mean recoveries were 93.6% for MDZ and 86.6% for 1-OHMDZ. Intra- and inter-day coefficient variations were less than 6.5 and 5.5% for MDZ, and 6.1 and 5.7% for 1-OHMDZ at 0.3, 4, 20 and 40ng/ml. The limits of quantification were 0.25ng/ml for both MDZ and 1-OHMDZ. This method was sensitive and reliable enough for pharmacokinetic studies on healthy volunteers, and was applied for the measurement of CYP3A activity in humans after an intravenous (1mg) and a single-oral administration (2mg) of subtherapeutic MDZ dose. PMID- 17085084 TI - Determination by liquid chromatography-mass spectrometry of clomiphene isomers in the plasma of patients undergoing treatment for the induction of ovulation. AB - A rapid, sensitive and selective LC-MS method is described for the simultaneous determination of zuclomiphene and enclomiphene in plasma from patients undergoing treatment with clomiphene citrate for the induction of ovulation. Samples spiked with N-didesmethyltamoxifen, the internal standard, were extracted into methyl tertiary butyl ether. The compounds were separated on a Luna C(18) analytical column, and a mobile phase of methanol-water (70:30 v/v) containing 0.05% trifluoroacetic acid at a flow rate of 1ml/min. The limits of determination were 35pg/ml and 7pg/ml for zu- and enclomiphene, respectively. Within-day coefficients of variation ranged from 2.1% to 7.2%. PMID- 17085085 TI - Analysis of hydroxyproline isomers and hydroxylysine by reversed-phase HPLC and mass spectrometry. AB - Collagens, the most abundant mammalian proteins, contain a high content of hydroxylated amino acids, such as, 3- and 4-cis-/trans-hydroxyproline (Hyp) and 5 hydroxylysine (Hyl). Whereas the global content of 4-Hyp was studied by amino acid analysis, no technique to determine all five hydroxyamino acids simultaneously in collagens has been reported. Here, we report the separation of all five hydroxyamino acids as well as two Hyp epimers from all other proteinogenic amino acids after derivatization with N(2)-(5-fluoro-2,4 dinitrophenyl)-l-valine amide (l-FDVA) by RPC-UV-ESI-MS. The general applicability of this method is shown for three Hyp-containing peptides as well as collagen type I. PMID- 17085086 TI - Proteomic analysis of isolated membrane fractions from superinvasive cancer cells. AB - The superinvasive phenotype exhibited by paclitaxel-selected variants of an in vitro invasive clonal population of the human cancer cell line, MDA-MB-435S were examined using DIGE (Fluorescence 2-D Difference Gel Electrophoresis) and mass spectrometry. Isolation of membrane proteins from the MDA-MB-435S-F/Taxol-10p4p and parental populations was performed by temperature-dependent phase partitioning using the detergent Triton X-114. Subsequent DIGE-generated data analysed using Decyder software showed many differentially-expressed proteins in the membrane fraction. 16 proteins showing statistically significant upregulation in the superinvasive cells were identified by MALDI-ToF. Proteins upregulated in the superinvasive population include Galectin-3, Cofilin, ATP synthase beta subunit, voltage-dependent anion channel 1, voltage dependent anion channel 2, ER 60 protein, MHC class II antigen DR52, Beta actin, TOMM40 protein, Enolase 1, Prohibitin, Guanine nucleotide-binding protein, Annexin II, Heat shock 70 kDa protein, Stomatin-like protein 2 and Chaperonin. Many of these proteins are associated with inhibition of apoptosis, the progression of cancer, tumourigenicity, metastasis, actin remodelling at the leading edge of cells, polarized cell growth, endocytosis, phagocytosis, cellular activation, cytokinesis, and pathogen intracellular motility. These results suggest a correlation between the increased abundance of these proteins with the superinvasive phenotype of the paclitaxel-selected MDA-MB-435S-F/Taxol-10p4p population. PMID- 17085087 TI - Bacterially expressed and optimized recombinant Phl p 1 is immunobiochemically equivalent to natural Phl p 1. AB - Recombinant production in bacteria of soluble and monomeric Phl p 1, a major allergen of Timothy grass pollen, has proved to be very problematic. In order to facilitate expression and purification of this allergen, a recombinant variant was designed with a single amino acid substitution. Several comparative analyses with natural counterparts using electrophoretic and HPLC separations, together with immunological assays, demonstrated high equivalence. This is the first description of an approach aiming at an improvement of a natural like recombinant allergen. PMID- 17085088 TI - Effect of nitinol wire surface properties on albumin adsorption. AB - The superelastic, shape memory alloy nitinol ( approximately 50% nickel and approximately 50% titanium) is an important medical device material used for stent applications. However, the role specific surfaces properties have in protein adsorption remain controversial. In this study the effects of nitinol wire surface roughness, hydrophobicity and elemental composition upon albumin adsorption are investigated. In particular, we demonstrate that the technique of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in the so-called surface mode can be used for the direct detection of albumin on the wire surfaces. In addition, albumin adsorbing to the wires was determined by using (125)I-labelled albumin. Albumin was detected on all wire samples. Surface roughness and hydrophobicity appeared to have no effect on albumin adsorption. There was however a clear correlation between the surface nickel and oxygen concentration and the amount of albumin adsorbed. Samples with higher levels of nickel and less oxygen in the surface oxide layer of the wires showed increased albumin adsorption, which could lead to improved biocompatibility. However, nickel is a toxic substance and can cause many adverse effects on humans, and thus nitinol with a slightly enriched surface nickel concentration that does not exhibit nickel release may have potential as a medical device material. PMID- 17085089 TI - Influence of thrombin concentration on the mechanical and morphological properties of cell-seeded fibrin hydrogels. AB - Fibrin is a biopolymer that has been used in a variety of biomaterial, cell delivery and tissue engineering applications. The enzyme thrombin catalyzes the formation of fibrin microfibrils, which form a three-dimensional mesh in which cells can be directly embedded at the time of gel formation. In this study, fibrin hydrogels containing vascular smooth muscle cells were created using varying concentrations of thrombin. Over 7 days in culture, all gels decreased in volume as the fibrin matrix compacted, and the degree of gel compaction increased as thrombin concentration decreased. The material modulus and ultimate tensile stress of the gels also increased with decreasing thrombin concentration. Addition of thrombin to similar constructs made using collagen Type I did not show an effect on gel compaction or mechanical properties, suggesting that these effects were a result of thrombin's action on fibrin polymerization, and not cellular functions. Cell proliferation in fibrin hydrogels was not significantly affected by thrombin addition. Matrix examination using scanning electron microscopy showed increasing fibrin fiber diameters as thrombin concentration decreased. Confocal microscopic imaging of the actin cytoskeleton showed that cell morphology on two-dimensional substrates of fibrin showed marked changes, with higher thrombin concentrations producing cells with longer cellular projections. However, these morphological changes were not as apparent in cells embedded in three-dimensional (3-D) matrices, in which cells exhibited a similar morphology independent of thrombin concentration. These results relate features of the matrix and cellular components of 3-D fibrin constructs to mechanical properties, and contribute to the understanding of structure-function relationships in cell-seeded, 3-D protein hydrogels. PMID- 17085090 TI - Development of glass-ceramic scaffolds for bone tissue engineering: characterisation, proliferation of human osteoblasts and nodule formation. AB - Glass-ceramic macroporous scaffolds for tissue engineering have been developed using a polyurethane sponge template and bioactive glass powders. The starting glass (CEL2) belongs to the system SiO(2)-P(2)O(5)-CaO-MgO-Na(2)O-K(2)O and has been synthesised by a conventional melting-quenching route. A slurry of CEL2 powder, polyvinyl alcohol and water has been prepared in order to coat, by impregnation, the polymeric template. An optimised thermal treatment was then use to remove the sponge and to sinter the glass powders, leading to a glass-ceramic replica of the template. Morphological observations, image analyses, mechanical tests and in vitro tests showed that the obtained devices are good candidates as scaffolds for bone-tissue engineering, in terms of pore-size distribution, pore interconnection, surface roughness, and both bioactivity and biocompatibility. In particular, a human osteoblast cell line (MG-63) seeded onto the scaffold after a standardised preconditioning route in simulated body fluid showed a high degree of cell proliferation and a good ability to produce calcium nodules. The obtained results were enhanced by the addition of bone morphogenetic proteins after cell seeding. PMID- 17085091 TI - VEGF and VEGF receptors are differentially expressed in chondrocytes. AB - During long bone development, cartilage replacement by bone is governed in part by angiogenesis. Although it has been demonstrated that vascular endothelial growth factor (VEGF-A) is crucial during endochondral ossification, little is known about the involvement of the other VEGF family members. Thus, we examined the expression and production of these members on primary chondrocytes and ATDC5 chondrogenic cells. VEGF-A, VEGF-B, VEGF-C and VEGF-D were shown to be expressed and synthesized demonstrating that numerous angiogenic factors can be produced by chondrocytes. In ATDC5 VEGF-A, VEGF-B and VEGF-C were over-expressed in the presence of chondrogenic and bone morphogenetic protein (BMP)-2 treatment suggesting that these factors play an important role during chondrogenesis. In addition, neuropilin-1, VEGF receptor-2 and VEGF receptor-3 gene expression were observed with an increase in VEGF-R2 expression under chondrogenic and BMP-2 treatment, suggesting that VEGF proteins could act in an autocrine/paracrine manner in addition to their angiogenic function. In conclusion, we demonstrated for the first time that chondrocytes secreted the four members of the VEGF family. We also showed that VEGF-B, VEGF-C and VEGF-D were secreted as processed proteins. The up-regulation of VEGF-B and VEGF-C at the mRNA and protein levels under chondrogenic stimulation strongly suggests a major role for these proteins in growth plate physiology. PMID- 17085092 TI - Neurotrophin-directed differentiation of human adult marrow stromal cells to dopaminergic-like neurons. AB - Marrow-isolated adult multilineage inducible (MIAMI) cells were differentiated in vitro to neuronal cells in a neurotrophin-dependent fashion. After induction, the cells revealed electrophysiological features similar to those observed in mature neurons. Primary early passage human MIAMI cells without any type of co-cultures with other cell types were used. The developmental program involved a multi-step process requiring the concerted action of brain-derived neurotrophic factor, nerve growth factor and depended on neurotrophin-3, after basic fibroblast growth factor withdrawal. MIAMI-derived neuron-like cells sequentially expressed the neuronal markers, developed a complex neurite outgrowth and arborization, and acquired electrophysiological characteristics similar to those observed in mature neurons. The young and old MIAMI-derived neuronal cells developed both inward and outward currents upon depolarization, similar to those observed in normal neurons. These results represent the earliest evidence that neurotrophin-3 can direct the differentiation of non-neural stem cells from human adult bone marrow stroma to neuron-like cells in vitro. Supplementing the aforementioned multi-step process with sonic hedgehog, fibroblast growth factor 8, and retinoic acid increased the expression of molecules involved in dopaminergic differentiation and of tyrosine hydroxylase, the rate limiting enzyme of dopamine synthesis. MIAMI cells from young and old individuals represent autologous human cell populations for the treatment of disorders of the skeletal and nervous systems and for applications in cell therapy and reparative medicine approaches. PMID- 17085096 TI - Choices. PMID- 17085097 TI - What's the trouble with cytology? PMID- 17085098 TI - Are there molecular signatures for predicting bladder cancer prognosis? PMID- 17085099 TI - Individualizing the approach to urological trauma. PMID- 17085100 TI - Male pelvic health: a urological call to arms. PMID- 17085101 TI - The epidemiology of renal cell carcinoma. AB - PURPOSE: We identified and examined risk factors for renal cell cancer, some of which may explain in part the trends of steadily increasing incidence rates, particularly in black Americans. MATERIALS AND METHODS: Epidemiological studies were identified through a MEDLINE(R) search of the literature through February 2006. A qualitative summary of the results of individual studies is presented. RESULTS: Cigarette smoking and obesity are the most consistently established causal risk factors, accounting for about 20% and 30% of renal cell cancers, respectively. Hypertension appears to independently influence renal cell cancer risk. Neither acetaminophen nor other analgesics have been convincingly linked with renal cell cancer. With respect to diet a general protective effect of fruit and vegetable consumption is the only consistently reported finding. For occupational factors the weight of the evidence provides no consistent support for the hypotheses that renal cell cancer may be caused by asbestos, gasoline or trichloroethylene exposure. Self-reported family history is associated with 2 to 3-fold increases in risk and the major inherited forms of renal cell cancer together account for about 2% of this malignancy. CONCLUSIONS: A further reduction in cigarette smoking, and a decrease in the rates of obesity and hypertension would likely moderate the increasing incidence of renal cell cancer. Epidemiological studies, including evaluation of gene-environment interactions, are needed to specifically identify reasons for the increasing incidence, particularly for assessing the roles of obesity and hypertension. Special attention should be focused on black Americans since their incidence rate recently increased to significantly surpass that in white Americans. PMID- 17085102 TI - Primary carcinoid tumors of the kidney. AB - PURPOSE: We describe in detail the features of carcinoid tumors of the kidney. We evaluated possible prognostic factors. MATERIALS AND METHODS: An extensive search was performed in the medical literature regarding primary carcinoid tumors of the kidney. Epidemiological, clinical, diagnostic, histopathological, therapeutic and prognostic data were evaluated. Several potential risk factors were compared with the incidence of metastases and clinical outcome of the patients. RESULTS: A total of 56 case reports were reviewed. Median patient age was 49 years. Horseshoe kidneys were present in 17.8% of cases. Incidental diagnosis was made in 28.6% of patients. The most common symptom was abdominal or flank pain and neuroendocrine syndromes occurred with only 12.7% of primary renal carcinoid tumors. Of the patients 73.6% presented with tumors larger than 4 cm. Metastases were present in 45.6% of patients at initial diagnosis and almost 60% with tumors greater than 4 cm had metastases. CONCLUSIONS: Renal carcinoid is the second most prevalent genitourinary carcinoid in each sex, following testicular carcinoids in men and ovarian tumors in women. Significant adverse prognostic factors include age greater than 40 years, tumor size greater than 4 cm, purely solid tumors on the cut surface, mitotic rate higher than 1/10 high power fields, metastasis at initial diagnosis and tumors extending throughout the renal capsule. PMID- 17085103 TI - The use of hemostatic agents and sealants in urology. AB - PURPOSE: While hemostatic agents and sealants have long been used in the fields of surgery and urology, confusion persists about their indications for use and the optimal agent choice. We comprehensively defined and evaluated the scientific basis for hemostatic agent and sealant use in urology, and provide a conceptual framework for future research and discussion. MATERIALS AND METHODS: A MEDLINE search of all available literature concerning hemostatic agents in urology was performed, including topical hemostats, anti-fibrinolytics, fibrin sealants and matrix hemostats. Select references were also chosen from the broader surgical literature. Animal studies, case reports, retrospective and prospective studies, and opinion articles were reviewed. RESULTS: Hemostatic agents include a wide range of components. Recent literature has focused on fibrin sealants and matrix agents. Two main indications exist for hemostatic agents, including 1) hemostasis and 2) sealant. The best evidence for efficacy and safety exists for hemostasis, especially for nephrectomy and trauma. Newer data highlight urinary tract reconstruction, fistula and percutaneous tract closure, suture line strengthening and infertility as potential uses. Novel drug delivery and tissue engineering are areas with large clinical potential. CONCLUSIONS: Hemostatic agent use is promising and yet unproven for most conditions currently treated in urology. Hemostasis continues to be the main indication, which is well established. Few trials have examined comparative efficacy among hemostatic agents and further prospective studies are needed to justify additional indications as well as determine the optimal mode of use. Minimally invasive surgery will further drive the use of hemostatic agents and sealants. Cost-effective, evidence based hemostatic agent use will continue to challenge all urologists. PMID- 17085104 TI - The application of botulinum toxin in the prostate. AB - PURPOSE: Botulinum neurotoxin is the most potent naturally occurring toxin known to inhibit various neurotransmitters. Injection of botulinum neurotoxin into the bladder and urethral sphincter has been used to treat bladder hyperactivity and sphincter dyssynergia. Recently botulinum neurotoxin application was extended to prostate disorders. Why would a urologist want to target the prostate? MATERIALS AND METHODS: We reviewed the literature on the mechanisms of action and clinical efficacy of botulinum neurotoxin treatment of the prostate. In addition to our personal clinical experience and basic research, information was gathered from MEDLINE and published abstracts from international meetings. We also present basic research and discuss the potential mechanism of action of botulinum neurotoxin on the prostate. RESULTS: There are 8 current peer reviewed publications on the injection of botulinum neurotoxin in the prostate. Cystoscopic transurethral or transperineal/transrectal ultrasound guided techniques have been used. Outcome improvement reported includes decreases in prostate size, prostate specific antigen and residual urine volume, and improvement in the flow rate and symptom score lasting 6 months or longer. CONCLUSIONS: Botulinum toxin has demonstrated exciting and promising preliminary results for male lower urinary tract symptoms. Translational research suggests novel mechanism of action of botulinum toxin in the prostate for benign prostatic hyperplasia and chronic nonbacterial prostatitis. It may even be considered as adjuvant treatment for prostate cancer. The use of botulinum neurotoxin in the prostate is currently Food and Drug Administration off label and in support of evidence based medicine practices caution should be applied until larger, randomized clinical studies are completed. More basic research is needed to identify the mechanisms by which botulinum toxin affects the prostate. PMID- 17085105 TI - The discovery of prostate specific antigen as a biomarker for the early detection of adenocarcinoma of the prostate. AB - PURPOSE: Prostate specific antigen is the most widely used oncological biomarker in medicine today. Before its implementation as an early diagnostic marker, urologists were limited to prostatic acid phosphatase, digital rectal examination and transrectal ultrasound for the detection of prostate cancer. We review the history of the discovery of prostate specific antigen as a biomarker for the early detection of adenocarcinoma of the prostate. MATERIALS AND METHODS: We performed a structured literature review, searching PubMed for papers on the subject of prostate specific antigen limited to humans between the years 1970 to 2005. We found a total of 8,365 articles. RESULTS: While the use of prostate specific antigen in evaluating newly diagnosed prostate disease, and followup of men after treatment for prostate disease is accepted practice, prostate specific antigen screening for prostate cancer remains controversial. CONCLUSIONS: In the next decade the results of randomized trials of screening may answer some of the questions posed at the beginning of the prostate specific antigen era. To what extent does prostate specific antigen screening affect prostate cancer mortality and at what cost? PMID- 17085106 TI - Variability in size measurement of renal masses smaller than 4 cm on computerized tomography. AB - PURPOSE: We assessed the degree of interobserver and intra-observer variability in the size measurement of small renal tumors with computerized tomography. MATERIALS AND METHODS: A total of 29 renal masses from 21 patients were reviewed independently by 3 radiologists on computerized tomography performed with 5 mm collimation and a reconstruction interval of 2.5 mm. Measurements were made of the largest axial dimension (X), perpendicular axial dimension (Y) and slice direction (Z) with double reads performed in a subset of patients. The predicted 95% CI was calculated for each measure, assuming different readers and the same reader. RESULTS: The predicted error bounds for a single renal mass measurement in the X axis were +/-3.1 mm when considering multiple readers and +/-2.3 mm for a single reader. In the X axis 78 of the 87 measurements (90%) were within 2 mm of the average measure. Smaller variances in measurements were seen with single reader repeat measurements than with multiple readers. Highest variances were seen in the Z axis. Measurements of volume and cross-sectional area showed a higher degree of variance. CONCLUSIONS: When comparing independent computerized tomography readings done with 5 mm collimation and a 2.5 mm reconstruction interval, a size change in the largest axial dimension of a renal mass of less than 3.1 mm between different radiologists and less than 2.3 mm for the same radiologist should be viewed with caution because it is within the range of measurement variability. PMID- 17085108 TI - Incidence of benign lesions for clinically localized renal masses smaller than 7 cm in radiological diameter: influence of sex. AB - PURPOSE: We determined the incidence of benign renal lesions in patients undergoing definitive surgery for localized renal masses 7 cm or less in maximum radiological diameter, and assessed preoperative and clinical parameters associated with benign histology. MATERIALS AND METHODS: The records of 1,184 patients who underwent consecutive partial or radical nephrectomies between January 2000 and January 2005 were retrospectively reviewed. We excluded 208 patients with lesions more than 7 cm in maximum radiological diameter, 17 with evidence of renal vein or vena caval invasion, 75 with suspected or documented metastatic disease, 28 with a history of renal cell carcinoma and 41 with no available imaging. Logistic regression was done to determine clinical factors associated with benign renal masses, including radiological tumor size, cystic vs solid appearance, patient sex, age, presenting symptoms and race. RESULTS: Of 815 nephrectomies in our data set 134 (16.4%) were associated with benign lesions, including oncocytoma in 87 (10.7%), angiomyolipoma in 17 (2%), simple cysts in 10 (1.2%), metanephric adenoma in 8 (1%), cystic nephroma in 5 (0.6%) and other in 7. On multivariate logistic regression analysis only sex was significantly associated with benign histology with females having an OR of 1.8 (95% CI 1.2 to 2.6, p = 0.002). Tumor size was not independently associated with benign histology (p = 0.13). CONCLUSIONS: A significant number (16.4%) of benign lesions less than 7 cm in radiological diameter were operated on based on suspicious preoperative imaging. Women had almost twice the likelihood of having a benign lesion. PMID- 17085111 TI - The evolving presentation of renal carcinoma in the United States: trends from the Surveillance, Epidemiology, and End Results program. AB - PURPOSE: The incidence of renal cancer is increasing, while cases series suggest that tumor size is decreasing. This has important implications for treatment planning. We evaluated national trends in renal cancer size and observed survival in patients diagnosed in the 3 periods 1988 to 1992, 1993 to 1997 and 1998 to 2002. MATERIALS AND METHODS: From the Surveillance, Epidemiology, and End Results database we identified 29,053 patients diagnosed with primary renal cancer. Patients were stratified into size categories and 5-year time cohorts. Size distribution was compared across cohorts. Kaplan-Meier survival curves and Cox proportional hazards modeling were used to examine trends in overall and stage specific survival. RESULTS: From 1988 through 2002 renal tumor size decreased from 66.8 to 58.6 mm, while the age adjusted incidence of renal cancer increased from 8.6 to 11.2 cases per 100,000 individuals. Kaplan-Meier analysis showed steadily deteriorating survival with increased cancer size above 4 cm with a median survival of 105 months for 4 to 7 cm vs 46 months for more than 7 cm. Cox modeling demonstrated significantly improved survival in patients diagnosed in the latter cohorts. With adjustment for size the latter cohorts remained significantly improved compared to the earliest cohort, although the 1998 to 2002 cohort was no longer significantly different than the 1993 to 1997 cohort. CONCLUSIONS: Nationally renal tumor size at presentation has steadily and consistently decreased. Patients more recently diagnosed had improved survival, which could be attributable to decreased tumor size in the latter cohorts. Patients more recently diagnosed also demonstrated a relative survival advantage independent of size compared to the earliest patients studied. PMID- 17085113 TI - Positive surgical parenchymal margin after laparoscopic partial nephrectomy for renal cell carcinoma: oncological outcomes. AB - PURPOSE: The oncological efficacy of partial nephrectomy is related to obtaining a negative surgical margin intraoperatively. This study assesses the oncological outcomes of patients undergoing laparoscopic partial nephrectomy for a renal tumor who had positive surgical margin on final pathology. MATERIALS AND METHODS: The experiences of 2 surgeons with 511 patients with a pathological diagnosis of renal cell carcinoma treated with laparoscopic partial nephrectomy were reviewed. Patients with a positive surgical margin were identified retrospectively. Oncological outcomes were assessed by followup with chest x-ray and computerized tomography every 6 to 12 months for 5 years. RESULTS: There were 9 patients (1.8%) with a positive margin on final pathology. Mean tumor size was 2.8 cm (range 1.7 to 4.0). Two patients underwent secondary completion radical nephrectomy, one at 4 days and the other at 2 months following laparoscopic partial nephrectomy. No residual tumor was identified in the nephrectomy specimen in either patient. Of the remaining 7 patients who elected surveillance, 1 with von Hippel-Lindau disease died of metastatic renal cell carcinoma to pancreas 10 months after laparoscopic partial nephrectomy. The remaining patients were disease-free for a median followup of 32 months (range 6 to 76). No patient in the series had port site seeding. CONCLUSIONS: A positive margin following laparoscopic partial nephrectomy does not necessarily indicate residual disease. However, vigilant monitoring is mandatory. While midterm outcomes parallel those of patients with a negative margin, longer followup is necessary to determine the ultimate oncological outcomes in this subgroup of patients. PMID- 17085114 TI - Are patients aware of the association between smoking and bladder cancer? AB - PURPOSE: Smoking is the single greatest risk factor for bladder cancer. Since few studies have demonstrated the efficacy of screening for bladder cancer, primary prevention by decreasing the modifiable risk factors is the best defense. An aspect of modifying a behavioral risk factor is awareness of the association between behavior and disease. While many anti-smoking campaigns specifically focus on lung cancer, few mention bladder cancer. We evaluated the awareness of smoking as a risk factor for bladder cancer. MATERIALS AND METHODS: Between February and May 2005 we prospectively surveyed patients presenting to a urology clinic regarding their knowledge of risk factors for bladder cancer and other cancers. The questionnaire also captured data regarding patient smoking habits. RESULTS: A total of 280 patients completed the survey, including 34% who were younger than 50 years, 63% who were male, 89% who were white and 57% who were college graduates. Only 36% vs 98% of the sample reported that smoking was a risk factor for bladder vs lung cancer. Patients with a higher level of education and females were statistically more likely to be aware of the association between smoking and bladder cancer. CONCLUSIONS: Patients at a urology clinic had low overall knowledge regarding bladder cancer risk factors. Most patients queried had no idea regarding the relationship between bladder cancer and tobacco use regardless of smoking status. Our study suggests the need for the American public to be better educated to help combat smoking related cancers. PMID- 17085117 TI - Is there an indication for frozen section examination of the ureteral margins during cystectomy for transitional cell carcinoma of the bladder? AB - PURPOSE: We evaluated the incidence of pathological findings of the ureter at cystectomy for transitional cell carcinoma of the bladder and assessed the usefulness of intraoperative frozen section examination of the ureter. MATERIALS AND METHODS: Histopathological findings of ureteral frozen section examination were compared to the corresponding permanent sections and the diagnostic accuracy of frozen section examination was evaluated. These segments were then compared to the more proximal ureteral segments resected at the level where they cross over the common iliac arteries. The histopathological findings of the ureteral segments were then correlated for upper urinary tract recurrence and overall survival. RESULTS: Transitional cell carcinoma or carcinoma in situ was found on frozen section examination of the distal ureter in 39 of 805 patients (4.8%) and on permanent sections in 29 (3.6%). In 755 patients the false-negative rate of frozen section examination of the ureters was 0.8%. Of the patients with carcinoma in situ diagnosed on the first frozen section examination 80% also had carcinoma in situ in the bladder. Transitional cell carcinoma or carcinoma in situ in the most proximally resected ureteral segments was found in 1.2% of patients. After radical cystectomy there was tumor recurrence in the upper urinary tract in 3% of patients with negative ureteral frozen section examination and in 17% with carcinoma in situ on frozen section examination. CONCLUSIONS: Routine frozen section examination of the ureters at radical cystectomy is only recommended for patients with carcinoma in situ of the bladder, provided the ureters are resected where they cross the common iliac arteries. PMID- 17085118 TI - Outcomes of radical cystectomy for transitional cell carcinoma of the bladder: a contemporary series from the Bladder Cancer Research Consortium. AB - PURPOSE: We present the characteristics and outcomes of a large, contemporary, consecutive series of patients treated with radical cystectomy and pelvic lymphadenectomy for transitional cell carcinoma of the bladder. MATERIALS AND METHODS: We developed a multi-institutional database and collected retrospective and prospective data on 888 consecutive patients with bladder transitional cell carcinoma who were treated with radical cystectomy and pelvic lymphadenectomy at 3 academic centers in the United States between 1984 and 2003. RESULTS: Of the patients 25% had extravesical tumor extension with negative lymph nodes and 23% had lymph node metastasis. The rate of lymph node involvement increased with advancing pathological stage. Mean recurrence-free and bladder cancer specific survival +/- SE was 58% +/- 2% and 66% +/- 2% at 5 years, respectively. On preoperative multivariate analysis clinical tumor stage and neoadjuvant systemic chemotherapy were associated with cancer recurrence, while more advanced age, clinical tumor stage and preoperative carcinoma in situ were associated with bladder cancer specific mortality. On postoperative multivariate analysis pathological tumor stage, lymph node metastasis, lymphovascular invasion, adjuvant radiotherapy and adjuvant chemotherapy were associated with cancer recurrence, while higher pathological tumor stage, more advanced age, lymph node metastasis, lymphovascular invasion and adjuvant radiotherapy were associated with disease specific survival. Patients with metastasis to regional lymph nodes (pT any N1-3) were at significantly higher risk for bladder cancer recurrence and death than patients with extravesical tumor extension (pT3N0), who in turn were at significantly higher risk than patients with organ confined disease (pT2 N0 or less). CONCLUSIONS: The results of this large, contemporary, multi-institutional series show that radical cystectomy and pelvic lymphadenectomy provide durable local control and disease specific survival in patients with localized invasive transitional cell carcinoma. PMID- 17085120 TI - Comparison of methods for calculating prostate specific antigen velocity. AB - PURPOSE: Prostate specific antigen velocity is frequently calculated using regression analysis of multiple prostate specific antigen measurements during an interval of 18 to 24 months. It has been reported that the prostate specific antigen velocity in the year before prostate cancer diagnosis is associated with the cancer specific mortality rate following radical prostatectomy and radiation therapy. There are limited data comparing alternate methods of calculating prostate specific antigen velocity. In this study we compared simple arithmetic prostate specific antigen velocity calculations to the more complicated regression analysis using prostate specific antigen measurements from varying intervals to determine whether the methods were interchangeable. MATERIALS AND METHODS: From 2003 to 2005 a total of 540 men underwent radical retropubic prostatectomy for localized prostate cancer. Preoperative prostate specific antigen velocity was calculated using arithmetic and linear regression methods during 12 and 18-month intervals. The correlation, mean and median prostate specific antigen velocity were compared among methods. RESULTS: Prostate specific antigen velocity calculations using simple arithmetic and linear regression had a strong correlation when restricted to prostate specific antigen values from 12 months before diagnosis (r = 0.92). When prostate specific antigen measurements beyond 1 year were included in the calculation, prostate specific antigen velocity was significantly lower than in the 12 months before cancer diagnosis. CONCLUSIONS: Using a simple arithmetic prostate specific antigen velocity calculation yields results comparable to those of the more complicated regression analysis when restricted to prostate specific antigen values from within the year before diagnosis in patients with clinically localized prostate cancer. In this manner arithmetic prostate specific antigen velocity calculations may be used in daily clinical practice to help predict the outcome following definitive therapy. Prostate specific antigen velocity is significantly lower when calculated during a longer interval before prostate cancer diagnosis. PMID- 17085122 TI - Dynamic contrast enhanced, pelvic phased array magnetic resonance imaging of localized prostate cancer for predicting tumor volume: correlation with radical prostatectomy findings. AB - PURPOSE: We assessed the value of pelvic phased array dynamic contrast enhanced magnetic resonance imaging for predicting the intraprostatic location and volume of clinically localized prostate cancers. MATERIALS AND METHODS: Suspicious areas on prospective pre-biopsy magnetic resonance imaging in 24 patients were assigned a magnetic resonance imaging malignancy score and located with respect to anatomical features, gland side, and transition and peripheral zone boundaries. The largest surface area and volume were measured. These magnetic resonance imaging findings were compared with radical prostatectomy specimen histopathology findings. RESULTS: Histopathology maps detected 56 separate cancer foci. The largest tumor focus was located in the peripheral zone in 14 patients and in the transition zone in 10. T1-weighted dynamic contrast enhanced magnetic resonance imaging identified 30 of the 39 tumor foci greater than 0.2 cc and 27 of the 30 greater than 0.5 cc. T2-weighted sequences were suspicious in 22 of 30 foci greater than 0.2 cc that were identified by T1-weighted dynamic contrast enhanced magnetic resonance imaging sequences. Sensitivity, specificity, and positive and negative predictive values for cancer detection by magnetic resonance imaging were 77%, 91%, 86% and 85% for foci greater than 0.2 cc, and 90%, 88%, 77% and 95% for foci greater than 0.5 cc, respectively. Median focus volume was 1.37 cc (range 0.338 to 6.32) for foci greater than 0.2 cc detected by magnetic resonance imaging in the peripheral zone and 0.503 cc (range 0.337 to 1.345) for those not detected by magnetic resonance imaging (p <0.05). Corresponding median values for transition zone foci were 2.54 (range 0.75 to 16.87) and 0.435 (range 0.26 to 0.58). CONCLUSIONS: Pre-biopsy pelvic phased array dynamic contrast enhanced magnetic resonance imaging is an accurate technique for detecting and quantifying intracapsular transition or peripheral zone tumor foci greater than 0.2 cc. It has promising implications for cancer detection, prognosis and treatment. PMID- 17085123 TI - Characteristics of patients with familial versus sporadic prostate cancer. AB - PURPOSE: There are conflicting reports concerning whether prostate cancer in families with multiple affected members has different clinical and pathological features than sporadic cases. In our study we compared the clinical characteristics, pathological outcomes and the 7-year biochemical progression free rate in patients with apparent sporadic prostate cancer, affected sibling pairs, families with multiple affected members and families meeting the Johns Hopkins criteria for hereditary prostate cancer. MATERIALS AND METHODS: From 1983 to 2003, 3,478 men underwent radical retropubic prostatectomy by a single surgeon (WJC). Of these men 1,186 reported family history status. We compared age at surgery, prostate specific antigen at diagnosis, pathological tumor stage, Gleason score, tumor characteristics and 7-year biochemical progression-free survival rates in the groups using chi-square, 1-way ANOVA or Cox proportional hazards regression analysis. RESULTS: The 7-year biochemical progression-free survival rates were 81% for sporadic cases, 71% for sibling pairs, 72% for hereditary cases and 81% for high density family members (p = 0.3). Of the clinical and pathological features examined only age (p <0.0001) and positive surgical margin rate (p = 0.03) were significantly different among groups. CONCLUSIONS: In our study population clinicopathological features and progression free survival are similar between sporadic and familial prostate cancer cases. The sibling pairs had a trend toward less favorable tumor features and progression-free survival, but the difference was not statistically significant. PMID- 17085125 TI - Impact of androgen deprivation therapy on physical and cognitive function, as well as quality of life of patients with nonmetastatic prostate cancer. AB - PURPOSE: Many patients with prostate cancer receive androgen deprivation therapy for long periods. We compared physical and cognitive function, and quality of life in a cross-sectional study of 57 patients receiving androgen deprivation therapy for nonmetastatic prostate cancer and 51 healthy age matched controls. MATERIALS AND METHODS: Physical and daily function were measured by the 6-minute walk test, grip strength, the timed up and go test, and activities of daily living measures. Cognitive function was evaluated by the High Sensitivity Cognitive Screen and by a self-report prototype Functional Assessment of Cancer Therapy cognitive function subscale. Quality of life was assessed by the Functional Assessment of Cancer Therapy-General with the subscale for fatigue and by the Patient-Oriented Prostate Utility Scale. RESULTS: Androgen deprivation therapy was given for a median of 1.8 years (range 0.4 to 7.4). Patients had lower median hemoglobin than controls (134 vs 148 gm/l, p <0.0001). Performance on physical tests was similar in the 2 groups. Moderate or severe cognitive impairment by the High Sensitivity Cognitive Screen was similar for patients (23%) and controls (35%, p = 0.2). Self-reported cognitive dysfunction was also similar. Scores for the Functional Assessment of Cancer Therapy-General were similar but the Patient Oriented Prostate Utility Scale summary score was worse for patients (median 71 vs 86, p <0.001). More patients reported severe fatigue (Functional Assessment of Cancer Therapy-Fatigue score less than 35, p = 0.03). Low energy, poor bladder control and loss of sexual function were reported in 36%, 47% and 95% of patients, and in 16%, 34% and 33% of controls, respectively. CONCLUSIONS: Patients treated with androgen deprivation therapy experience more symptoms and have worse fatigue than controls, but this study did not detect any effect on physical or cognitive function. PMID- 17085126 TI - Age stratified functional outcomes after laparoscopic radical prostatectomy. AB - PURPOSE: We assessed health related quality of life in men treated for prostate cancer with laparoscopic radical prostatectomy. MATERIALS AND METHODS: Urinary continence and potency were prospectively assessed in 369 men stratified into age groups, including group 1-younger than 50, group 2-50 to 59 and group 3-60 years or older, by analyzing answers to the Expanded Prostate Cancer Index Composite questionnaire collected preoperatively, and 3, 6 and 12 months after laparoscopic radical prostatectomy. RESULTS: Younger men were more likely to achieve urinary continence (1 pad or less daily) 1 year after laparoscopic radical prostatectomy (groups 1 to 3 100%, 91% and 81%, respectively, p <0.01). Younger men were also more likely to be potent and engaging in intercourse 1 year after bilateral nerve sparing laparoscopic radical prostatectomy (groups 1 to 3 70%, 67% and 46%, respectively, p <0.01). The mean percent return to baseline Expanded Prostate Cancer Index Composite urinary continence subscale at 1 year in groups 1 to 3 was 80%, 79% and 74%, respectively (p = 0.49). The mean percent return to baseline Expanded Prostate Cancer Index Composite sexual function subscale at 1 year in groups 1 to 3 was 68%, 65% and 58%, respectively (p = 0.56). Binary logistic regression modeling using the variables age group, prostate weight, International Prostate Symptom Score and nerve sparing status demonstrated that only younger age group was associated with return to continence at all postoperative time points (p <0.05). Younger age group was associated with return to early potency at 3 and 6 months (p = 0.02 and <0.01, respectively). However, only nerve sparing status was associated with recovery of potency at all time points (p <0.05). CONCLUSIONS: Younger men treated with nerve sparing laparoscopic radical prostatectomy regain urinary control and potency earlier than older men. However, validated questionnaire subscale analyses demonstrated that the return to preoperative baseline urinary continence and sexual function is similar in all age groups by the end of postoperative year 1. PMID- 17085127 TI - Is biopsy Gleason score independently associated with biochemical progression following radical prostatectomy after adjusting for pathological Gleason score? AB - PURPOSE: Biopsy Gleason score is known to be associated with prostate specific antigen failure following radical prostatectomy. However, it is unclear whether it remains associated with outcome after surgery when the pathological Gleason score is known. MATERIALS AND METHODS: We determined the association between biopsy Gleason score and biochemical progression after correcting for preoperative and postoperative characteristics, including pathological Gleason score, in 1,931 men treated with radical prostatectomy between 1988 and 2005 in the Shared Equal Access Regional Cancer Hospital Database Study Group database. Gleason score was examined as a categorical variable of 2 to 6, 3 + 4 and 4 + 3 or greater. RESULTS: Higher biopsy Gleason scores were positively associated with extracapsular extension (p <0.001), positive surgical margins (p <0.001), seminal vesicle invasion (p <0.001), positive lymph nodes (p <0.001) and biochemical progression (log rank p <0.001). After adjusting for only preoperative characteristics biopsy Gleason 3 + 4 and 4 + 3 or greater were associated with increased risk of biochemical progression compared to biopsy Gleason 6 or less (p = 0.001 and <0.001, respectively). After further adjusting for multiple pathological characteristics, including pathological Gleason score, the association between higher biopsy Gleason score and progression was little changed, in that men with biopsy Gleason 3 + 4 and 4 + 3 or greater were significantly more likely to experience progression (p = 0.001 and <0.001, respectively). Furthermore, when stratified by pathological Gleason score, higher biopsy Gleason scores were associated with an increased risk of biochemical progression in each pathological Gleason score category (log rank p 0.05). CONCLUSIONS: Our data indicate that bilateral varicocelectomy (for clinically palpable bilateral varicoceles) is associated with a significantly greater improvement in sperm quality (motility) and male fertility potential than unilateral varicocelectomy. The greater improvement in semen parameters and pregnancy outcome associated with bilateral varicocelectomy support the concept of a detrimental dose-effect of varicocele on male fertility potential. PMID- 17085172 TI - Discharge practice patterns following cystectomy for bladder cancer: evidence for the shifting of the burden of care. AB - PURPOSE: Between 1985 and 2000 the incidence of bladder cancer in this country increased by 33%. Radical cystectomy is the primary treatment modality in patients with invasive disease. We characterized trends in cystectomy use and discharge practice patterns following cystectomy during a recent 13-year period. MATERIALS AND METHODS: The Nationwide Inpatient Sample comprises a 20% probability sample of hospital discharges in the United States each year. Patients who underwent cystectomy for bladder cancer from 1988 to 2000 were identified using International Classification of Disease, 9th Revision, Clinical Modification codes. Outcome variables included in-hospital mortality, length of stay and hospital discharge status. RESULTS: From 1988 to 2000 an estimated 119,491 patients underwent cystectomy for bladder cancer. Cystectomy rates decreased by 17% from 3.91/100,000 (1988 to 1990) to 3.25/100,000 (1997 to 2000), largely due to a decreasing number of partial cystectomies. In-hospital mortality decreased from 3.3% (1988 to 1990) to 2.5% (1997 to 2000) (p = 0.027). Similarly median length of stay decreased from 13 days (1988 to 1990) to 9 (1997 to 2000) (p <0.001). During the same period the percent of patients being discharged to subacute care facilities increased from 5.3% to 13.2% (p <0.001), as did the percent of patients requiring home health care services (24.1% to 38.7%, p <0.001). CONCLUSIONS: From 1988 to 2000 there was a decrease in the rate of cystectomies being performed for bladder cancer with a substantial decrease in partial cystectomy use. During this period short-term outcomes following cystectomy improved, while the use of subacute care facilities and home health services increased dramatically, underscoring a shift in the burden of care in this patient population. PMID- 17085174 TI - A multi-institution, minimally invasive urological oncology fellowship: a critical assessment of the clinical training and academic benefits. AB - PURPOSE: We evaluated the clinical training and academic productivity of a unique minimally invasive urological oncology fellowship performed in 3-month rotations at 4 institutions. MATERIAL AND METHODS: With Georgia Cancer Coalition grant funding and institutional support a faculty urologist (JAB) completed 3-month fellowships at Thomas Jefferson University, Philadelphia in 2002, Indiana University, Indianapolis in 2003, Massachusetts General Hospital, Boston in 2003 and Henry Ford Hospital, Detroit in 2004. RESULTS: The trainee operated under the direction of 8 surgeons and assisted/observed another 5. Total operative experience was 355 cases, including 53 standard laparoscopic radical prostatectomies, 100 robotic assisted laparoscopic radical prostatectomies, 30 standard (including 13 donor) and 22 hand assisted laparoscopic nephrectomies, 6 nephroureterectomies, 14 partial nephrectomies, 3 renal cyst decortications, 12 pyeloplasties, 5 adrenalectomies, 2 hand assisted laparoscopic ureterolysis procedures, 1 laparoscopic partial and 1 radical cystectomy, hand assisted laparoscopic cystectomy, robotic cystectomy, 26 open and 2 laparoscopic retroperitoneal lymph node dissections, 5 complex open bladder surgeries, 6 complex open renal surgeries and approximately 24 endoscopic laser upper tract tumor cases. Post-fellowship sequential initiation of laparoscopic renal cancer (April 2002), prostatectomy (July 2003) and donor nephrectomy (November 2003) programs was accomplished at the home institution. Academic projects were completed during each fellowship phase with 43 presented abstracts and 2 book chapters, 2 non-peer reviewed articles and 12 peer reviewed articles published to date. CONCLUSIONS: A multi-institution fellowship allows serial acquisition and incorporation of a wide variety of cutting edge, minimally invasive and oncological procedures into an academic practice. It allows greater exposure to more high volume experts in varying oncological subspecialties. Clinical research and academic productivity are possible. PMID- 17085176 TI - Abstracts presented at the American Urological Association Annual Meeting: determinants of subsequent peer reviewed publication. AB - PURPOSE: Abstracts submitted to medical meetings do not undergo the same critical peer review process as published manuscripts. Despite this limited scrutiny presented abstracts often influence clinical thinking and practice. Consequently the peer reviewed publication rate of abstracts becomes critical in judging the quality of this research. We determined this publication rate and factors influencing it. MATERIALS AND METHODS: All 1,584 abstracts presented at the 2000 American Urological Association Annual Meeting were reviewed and assessed for subsequent publication with a fixed MEDLINE search protocol. We searched for publications from January 1, 1999 to May 31, 2005. Abstracts were deemed published if 1) at least 1 author of the presented abstract was a manuscript author and 2) at least 1 conclusion in the presented abstract was included in the final publication conclusions. Publication rates according to mode and topic of presentation, country or state of origin and time to publication were calculated. Journal impact factors for publications were compared according to these variables. RESULTS: Of presented abstracts from the 2000 American Urological Association meeting 55% went on to successful publication, including 59% of podium, 55% of poster, 55% of unmoderated poster and 42% of video presentations. Mean time from presentation to publication was 17 months. The average journal impact factor was 3.2. CONCLUSIONS: A significant proportion of presentations at the American Urological Association Annual Meeting is never subjected to or fails the critical peer review process. The overall journal impact factor for published manuscripts is modest. Meeting attendees should consider these observations when deciding whether to incorporate the findings of presentations into their clinical practice. PMID- 17085178 TI - Vaginal discharge and bleeding in girls younger than 6 years. AB - PURPOSE: Persistent unexplained vaginal discharge or bleeding in the pediatric population may be the only manifestation of a serious underlying medical or social problem. Therefore, these symptoms require careful and complete evaluation to identify the primary pathology accurately. We retrospectively reviewed charts of patients who presented for evaluation of persistent vaginal discharge or bleeding to determine if noninvasive imaging was a sensitive means of screening for gynecological pathology. MATERIALS AND METHODS: The records of 24 girls younger than 6 years who presented with vaginal discharge or bleeding were reviewed retrospectively. All patients were evaluated with noninvasive imaging, a pelvic examination while under anesthesia, vaginoscopy and cystoscopy. RESULTS: Noninvasive imaging was useful in identifying 5 of 7 vaginal foreign bodies. However, noninvasive imaging identified only 2 of 6 malignancies. These malignancies consisted of rhabdomyosarcoma (3 patients) and endodermal sinus tumor (3). Two girls also had benign vaginal mullerian papillomas that were not identified by noninvasive imaging. Noninvasive imaging did not aid in the diagnosis of sexual abuse. CONCLUSIONS: Based on these data, we recommend that all girls younger than 6 years who present with persistent vaginal discharge or bleeding be evaluated with pelvic examination while under anesthesia, to be followed by vaginoscopy and cystoscopy if no readily identifiable pathology is found by simple genital examination alone, regardless of the results of noninvasive imaging studies. PMID- 17085179 TI - Transperitoneal and retroperitoneal laparoscopic heminephrectomy--what approach for which patient? AB - PURPOSE: We report our experience with laparoscopic heminephrectomy using the transperitoneal and retroperitoneal approaches in 48 pediatric patients. MATERIALS AND METHODS: A total of 48 laparoscopic heminephrectomies were performed in 35 girls and 13 boys 45 days to 17 years old (mean 4.08 years) between September 1998 and March 2005. The procedures consisted of 44 upper pole heminephrectomies with partial or total ureterectomies and 4 lower pole heminephroureterectomies. Surgeries were performed using a transperitoneal approach in 32 patients (67%) and a retroperitoneal approach in 16 (33%). RESULTS: Followup ranged from 0.75 to 7.25 years (mean 3.53). In the retroperitoneal group 2 procedures required conversion, 1 to open heminephrectomy and 1 to a transperitoneal laparoscopic approach. Complications were seen in 5 of 48 patients (10%). Complications in the retroperitoneal group were seen in 2 patients. One patient had a postoperative urinary leak that resolved spontaneously. Another patient had development of a urinoma that was treated conservatively. Complications in the transperitoneal group were seen in 3 patients. One patient required an intraoperative chest tube due to pneumothorax, 1 had recurrent urinary tract infection that required excision of a short ureteral remnant and 1, 6-month-old boy had development of postoperative hypertension. Four of the 5 complications (80%) were seen in patients younger than 1 year. CONCLUSIONS: Transperitoneal and retroperitoneal laparoscopic heminephrectomy can be performed for benign disease in children with minimal morbidity, improved cosmesis and short hospital stay. Complication rate does not depend on the surgical approach, but rather on the age of the patient. PMID- 17085181 TI - Laparoscopic ureteral reimplantation with extracorporeal tailoring for megaureter: a simple technical nuance. AB - PURPOSE: We describe a novel technique of extracorporeal tailoring for megaureter to perform laparoscopic extravesical transperitoneal ureteral reimplantation. We present our initial results with this procedure. MATERIALS AND METHODS: Three cases of megaureter were operated on laparoscopically. The 3-port technique was used (all 5 mm in 2 cases, and 2, 5 mm and 1, 10 mm in 1). The colon was reflected medially to expose the retroperitoneal course of the ureter. The ureter was gently dissected circumferentially down to the bladder. It was ligated close to the bladder and divided proximal to it. The free ureteral end was delivered out through the ipsilateral 5 mm port. The lower end was tailored over an 8Fr feeding tube. A 6Fr Double-J stent was placed. Later the whole assembly was carefully replaced in the abdomen. A Lich-Gregoir type extravesical reimplantation was done. RESULTS: Mean operating time was 220 minutes (range 210 to 240), which included 15 to 20 minutes to exteriorize the ureter, complete extracorporeal tailoring and replace the stented ureter in the abdomen. Mean blood loss was 40 ml. Cystourethrogram at 3 months did not demonstrate any reflux. Renal scan showed preserved renal function in all 3 cases, with prompt drainage in 2 and delayed drainage in 1. At a mean followup of 12 months all 3 patients were asymptomatic. CONCLUSIONS: Extracorporeal tailoring for obstructing megaureter is an easy and safe procedure that makes laparoscopic ureteral reimplantation simpler and quicker to perform. PMID- 17085182 TI - Can a complete primary repair approach be applied to cloacal exstrophy? AB - PURPOSE: Surgical reconstruction for children with cloacal exstrophy remains challenging. The operative approach to cloacal exstrophy has expanded with the addition of the complete primary exstrophy repair. We assessed the safety and efficacy of complete primary exstrophy repair for this complex condition. MATERIALS AND METHODS: We performed a retrospective review of children treated from birth for cloacal exstrophy between March 1, 1994 and January 1, 2003 at our institution. We evaluated associated anomalies, method of closure, complications and urinary continence. RESULTS: Seven patients with cloacal exstrophy were initially treated at our institution. One mortality occurred before complete primary exstrophy repair was attempted. Six patients were converted to a classic exstrophy appearance and underwent closure using complete primary exstrophy repair principles within 7 to 182 days (mean 68, median 32) postoperatively. Postoperative development of moderate hydronephrosis was seen in 1 patient and severe hydronephrosis in 1. Three of six patients had vesicoureteral reflux. Six patients had dry intervals and spontaneous voids before toilet training. Two patients had stress urinary incontinence. Two patients have been treated with bladder neck injections. One has undergone bladder neck reconstruction and construction of a nonorthotopic channel for clean intermittent catheterization (Mitrofanoff). One patient reported complete dryness after toilet training. One child has undergone bladder augmentation. CONCLUSIONS: This series represents our initial efforts to use complete primary exstrophy repair for cloacal exstrophy. The application of the principles of complete primary exstrophy repair in a sequential fashion appears to be a viable and safe addition to the surgical armamentarium in this challenging patient population. PMID- 17085183 TI - The cost-effectiveness of dextranomer/hyaluronic acid copolymer for the management of vesicoureteral reflux. 2. Reflux correction at the time of diagnosis as a substitute for traditional management. AB - PURPOSE: The use of dextranomer/hyaluronic acid copolymer has become increasingly popular as an alternative to ureteral reimplantation in the treatment of vesicoureteral reflux. We compared the cost-effectiveness of performing dextranomer/hyaluronic acid injection at the time of diagnosis of reflux to that of traditional management. MATERIALS AND METHODS: A model to estimate the costs of managing vesicoureteral reflux has previously been created. We updated the model to compare the costs of managing vesicoureteral reflux by traditional methods with the costs of managing reflux if dextranomer/hyaluronic acid injection is performed at the time of diagnosis. The success rate required for dextranomer/hyaluronic acid injection at diagnosis to be as cost effective as traditional management was estimated. We used 2 models of dextranomer/hyaluronic acid injection at diagnosis-injection at diagnosis proceeding to traditional management if injection failed (scenario 1), and injection at diagnosis proceeding to ureteral reimplantation if injection failed (scenario 2). RESULTS: If reflux is stratified by grade in scenario 1, for grades III, IV and V respective success rates of 88.5%, 66.6%, and 55.6% for unilateral reflux and 97.5%, 89.7% and 91.4% for bilateral reflux must be achieved to have equal cost effectiveness to traditional management, while grades I and II reflux can never achieve equal cost-effectiveness. Stratified by grade for scenario 2, for grades III, IV and V respective success rates of 86.9%, 70.8% and 55.8% for unilateral reflux, and 97.6%, 89.8% and 89.8% for bilateral reflux must be achieved to attain equal cost-effectiveness compared to traditional management. In scenario 2 dextranomer/hyaluronic acid injection at diagnosis for grades I and II unilateral and bilateral reflux can never achieve equal cost-effectiveness compared to traditional management. CONCLUSIONS: Based on the results of this study, in most clinical situations dextranomer/hyaluronic acid injection at the time of diagnosis is unlikely to be as cost effective as traditional management of vesicoureteral reflux. PMID- 17085186 TI - Lack of usefulness of positioned instillation of contrast cystogram after injection of dextranomer/hyaluronic acid. AB - PURPOSE: Positioned instillation of contrast cystograms have been touted as possibly being more sensitive than standard cystograms for evaluation of vesicoureteral reflux. We performed positioned instillation of contrast cystograms intraoperatively, immediately after the injection of dextranomer/hyaluronic acid to treat vesicoureteral reflux, to determine whether they might be predictive of operative success and obviate the need for the standard postoperative voiding cystourethrogram, which is usually performed at 3 months. MATERIALS AND METHODS: Patients with vesicoureteral reflux and no confounding conditions were treated with dextranomer/hyaluronic acid and subsequent positioned instillation of contrast cystogram while under the same anesthesia between November 2003 and March 2005. The results of this intraoperative cystogram were compared to the results of the postoperative voiding cystourethrogram performed 3 to 4 months later. RESULTS: A total of 61 patients met the inclusion criteria and underwent positioned instillation of contrast cystogram after dextranomer/hyaluronic acid injection. Only 53 patients (86 ureters) completed the necessary postoperative evaluation. Positioned instillation of contrast cystogram added 4 minutes to the procedure and required about 4 seconds of fluoroscopy per ureter evaluated. The overall success rate for correcting reflux was 84% (72 of 86 ureters cured). None of the 14 ureters with persistent postoperative reflux was identified by intraoperative cystogram, and 3 patients were misidentified as having reflux despite cure confirmed postoperatively. Intraoperative positioned instillation of contrast cystogram was predictive of treatment failure 0% of the time (sensitivity 0%). There were no complications. CONCLUSIONS: Positioned instillation of contrast cystogram performed immediately after injection of dextranomer/hyaluronic acid was not useful in predicting which patients would have persistent reflux postoperatively. Patients are best served with the extant protocol of conventional cystography 3 to 4 months postoperatively. PMID- 17085187 TI - Endoscopic treatment of vesicovasal and vesicoureteral reflux in infants with persisting mesonephric duct. AB - PURPOSE: We present a minimally invasive endoscopic approach for the treatment of persisting mesonephric duct in male infants. MATERIALS AND METHODS: Five male infants 2 to 8 months old were referred with a history of recurrent sepsis, epididymo-orchitis and anorectal malformation that was treated elsewhere with initial colostomy. All infants had edematous unilateral testes, fever and poor feeding. Urine and blood cultures yielded the same microorganisms. All infections occurred while the infants were on regimens of prophylactic antibiotics. Comprehensive urological evaluations confirmed ipsilateral renal agenesis, sacral hypoplasia (3 patients), high anorectal malformation, and vesicovasal and vesicoureteral reflux. RESULTS: All 5 infants underwent urethrocystoscopy under general anesthesia. The ectopic persisting mesonephric duct entered the bladder neck and proximal prostatic urethra. Injection of 0.4 to 0.7 ml Urocol, used as a bulking agent, was administered submucosally at the opening of the persisting mesonephric duct. In 1 patient the orifice of the anomalous duct was not found during urethrocystoscopy, and ipsilateral vas ligation by titanium clips was performed. The results in the remaining 4 patients were excellent, and no further episodes of epididymitis were observed during a mean followup of 30 months. CONCLUSIONS: The diagnosis of persisting mesonephric duct should be considered in a male child with anorectal anomalies and recurrent epididymo-orchitis, and can be confirmed by radiological studies and cystoscopy. Endoscopic treatment of this anomaly should be considered first line therapy in these patients. However, surgical intervention is mandatory for children not responding to the procedure. PMID- 17085188 TI - Miniature intravesical urethral lengthening procedure for treatment of pediatric neurogenic urinary incontinence. AB - PURPOSE: Resistance to flow in a fluid conduit is proportional to tube length divided by the radius to the fourth power (Poiseuille's law). We report the results of a miniature intravesical urethral lengthening procedure where outlet resistance is increased by minimizing the diameter of the intravesical urethral tube. MATERIALS AND METHODS: Nine pediatric patients with preoperative intractable incontinence underwent the miniature intravesical urethral lengthening procedure along with continent catheterizable stoma (9 patients) and bladder augmentation (8). The intravesical portion of the urethral lengthening was 3 cm (traditionally 6 cm), and the urethra was tubularized around an 8Fr feeding tube (traditionally a 16Fr catheter). After the tubularized caudal portion was tunneled under the trigone the cephalad part of the urethra was placed as an onlay to the posterior bladder wall without ureteral reimplantation. RESULTS: At a mean followup of 31 months (range 10 to 47) 8 patients reported dry intervals of 3 hours or more, with minor leak per urethra only if they were overdue on the catheterization schedule. Mean postoperative abdominal leak point pressure was 71 cm H(2)O (range 28 to 116). Upper tracts were well preserved in all patients. One patient required bladder neck closure for intractable incontinence. CONCLUSIONS: The miniature intravesical urethral lengthening procedure requires minimal bladder tissue and is easy to perform. It appears to be an effective alternative in bladder neck reconstructive techniques, avoiding the need for ureteral reimplantation due to its small size, while functioning as a pop-off valve when the bladder is overly full. This procedure should be avoided in patients who lack a trigonal bar. PMID- 17085190 TI - Clinical and molecular markers of chronic interstitial nephropathy in congenital unilateral ureteropelvic junction obstruction. AB - PURPOSE: We evaluated clinical and biological variables, and their meaning as reliable markers of chronic interstitial nephropathy in a selected group of children with prenatally detected hydronephrosis who underwent pyeloplasty because of congenital unilateral ureteropelvic junction obstruction. MATERIALS AND METHODS: We reviewed the clinical, prenatal and postnatal ultrasonographic, and scintigraphic records of children for whom intraoperative biopsy records were available. We performed histological analysis, and evaluated tubulointerstitial immunostaining for vimentin and alpha-smooth muscle actin, and the immunohistochemical and mRNA expression of the renin-angiotensin system peptides and transforming growth factor-beta1. RESULTS: The children were divided in 2 groups according to the absence (group 1) or presence (group 2) of chronic interstitial nephropathy in the biopsy. Patients in group 2 were significantly younger at prenatal diagnosis (p = 0.031), and had decreased split renal function (p = 0.005) and worse drainage (p = 0.035) on preoperative diuretic renography. No differences were found in terms of degree of hydronephrosis, or its prenatal and postnatal variation. Group 2 biopsies exhibited greater immunostaining for alpha-smooth muscle actin and vimentin (p = 0.004 and p = 0.047, respectively), and transforming growth factor-beta1 mRNA levels (p = 0.06). Vimentin and alpha smooth muscle actin positivity correlated with renin, angiotensin II receptors 1 and 2, and transforming growth factor-beta1 mRNA levels, and all correlated with preoperative split renal function and post-void washout. CONCLUSIONS: In congenital unilateral ureteropelvic junction obstruction chronic interstitial nephropathy and poor postoperative recovery seem to be associated with an earlier diagnosis of hydronephrosis, functional loss greater than 10% and worse scintigraphic drainage. Moreover, there is a strong correlation between molecular fibrogenic markers and histologically and scintigraphically demonstrated renal damage. PMID- 17085194 TI - Testicular vasculitis. PMID- 17085193 TI - Gender specific chronological and morphometric assessment of fetal bladder wall development. AB - PURPOSE: To enhance our understanding of sonographically visible alterations in bladder wall thickness, we delineated phenotypic changes occurring in developing smooth muscle cells of the fetal and postnatal bladder with respect to gender specific differences. MATERIALS AND METHODS: Bladders of 30 male and 18 female fetuses and 4 stillborn infants were immunostained with an alpha-smooth muscle actin antibody. Morphological and morphometric assessment was performed with the assistance of an image analysis system. RESULTS: Alpha-smooth muscle actin expression in fetal bladder wall was detectable at 9 weeks of gestation. Bladder wall thickness and mean profile area of smooth muscle bundles increased significantly with advancing gestation, mediated by linear growth patterns. Fetal bladder wall development occurred uniformly, unrelated to gender. CONCLUSIONS: Although the lower urinary tract emerges in a gender specific way, our results suggest that in normal fetal growth detrusor muscle formation proceeds independent of genital sex. PMID- 17085195 TI - Computerized tomography criteria for the diagnosis of high grade papillary urothelial carcinoma. PMID- 17085196 TI - Molecular profiling of bladder tumors based on the detection of FGFR3 and TP53 mutations. AB - PURPOSE: On a routine basis we performed systematic molecular screening for FGFR3 and TP53 mutations in 121 bladder tumors. We then specifically analyzed the predictive value of the recurrence of FGFR3 and TP53 genotypes in superficial lesions. MATERIALS AND METHODS: The FGFR3 gene was analyzed by direct sequencing of exons 7, 10 and 15, whereas TP53 status was determined using the p53 functional assay in yeast. RESULTS: We identified a missense FGFR3 mutation in 66% of pTa, 26% of pT1 and 12% of pT2 tumors. Of activating FGFR3 mutations 54% and 85% were found in low G1 and intermediate G2 grade tumors, respectively, but in only 20% of high grade G3 tumors. We detected inactivating TP53 mutations in 10% of pTa, 42% of pT1 and 58% of pT2 tumors. Moreover, TP53 mutations were found only in 23% of grade G1 and 3% of grade G2 tumors but in 44% of high grade G3 tumors. When the 2 genotypes were combined, we observed that 58% of pTa tumors had the (mutant FGFR3, WT TP53) genotype, whereas 58% of invasive lesions harbored the inverse genotype (WT FGFR3, mutant TP53). The (mutant FGFR3, WT TP53) genotype and the (WT FGFR3, mutant TP53) genotype were detected in 23% and 38% of pT1G3 tumors, respectively. In the subgroup of 92 patients with superficial pTa-T1 bladder tumors we did not find that the TP53 or FGFR3 genotype alone or combined had a predictive value for tumor recurrence. CONCLUSIONS: Our data again represent solid proof for the pivotal role of FGFR3 and TP53 mutations in superficial and invasive bladder tumors, respectively. However, other molecular markers should be identified for borderline pT1G3 bladder tumors, which are probably at the crossroads of these 2 distinct molecular pathways. PMID- 17085197 TI - Immunotherapeutic effect of Concholepas hemocyanin in the murine bladder cancer model: evidence for conserved antitumor properties among hemocyanins. AB - PURPOSE: We determined the antitumor properties of a newly available hemocyanin obtained from the Chilean gastropod Concholepas concholepas (Biosonda Corp., Santiago, Chile) in a syngeneic heterotopic mouse bladder carcinoma model. Since keyhole limpet hemocyanin (Pierce, Rockford, Illinois) is used increasingly in biomedicine as a carrier for vaccines and an immunotherapeutic agent for bladder transitional cell carcinoma, there is a growing interest in finding new substances that share its potent immunomodulatory properties. Considering that keyhole limpet hemocyanin and Concholepas concholepas hemocyanin differ significantly, it was not possible to predict a priori the antitumor properties of Concholepas concholepas hemocyanin. MATERIALS AND METHODS: C3H/He mice were primed with Concholepas concholepas hemocyanin before subcutaneous implantation of mouse bladder tumor-2 cells. Treatment consisted of a subcutaneous dose of Concholepas concholepas hemocyanin (1 mg or 100 mug) at different intervals after implantation. Keyhole limpet hemocyanin and phosphate buffered saline served as positive and negative controls, respectively. In addition, experiments were designed to determine which elements of the immune response were involved in its adjuvant immunostimulatory effect. RESULTS: Mice treated with Concholepas concholepas hemocyanin showed a significant antitumor effect, as demonstrated by decreased tumor growth and incidence, prolonged survival and lack of toxic effects. These effects were similar to those achieved with keyhole limpet hemocyanin. We found that each hemocyanin increased natural killer cell activity but the effect of Concholepas concholepas hemocyanin was stronger. Analysis of serum from treated mice showed an increased interferon-gamma and low interleukin 4, which correlated with antibody isotypes, confirming that hemocyanins induce a T helper type 1 cytokine profile. CONCLUSIONS: To our knowledge our results are the first demonstration of the antitumor effect of a hemocyanin other than keyhole limpet hemocyanin. They suggest that this is an ancient conserved immunogenic mechanism shared by those hemocyanins that is able to enhance T helper type 1 immunity and lead to antitumor activity. Therefore, Concholepas concholepas hemocyanin may be an alternative candidate for providing safe and effective immunotherapy for human superficial bladder cancer. PMID- 17085198 TI - Radiation induced late damage to the barrier function of small blood vessels in mouse bladder. AB - PURPOSE: We identified changes in vascular barrier function in relation to collagen deposition during the late radiation response of mouse bladders. In this study albumin leakage was assessed as a marker of blood vessel barrier disruption. MATERIALS AND METHODS: Female C3H/Neu mice were irradiated with a single dose of 20 Gy and sacrificed for (immuno) histological studies after 90, 120, 180, 240 and 360 days. The fractional area of subepithelial tissue with extravascular albumin signal was defined. Moreover, the amount of collagen was determined after Masson's trichrome staining on an arbitrary score of 0 to 3. RESULTS: Subepithelial albumin infiltration due to leakage from the blood vessels was seen in irradiated animals during the entire late phase. It was most pronounced at days 120 and 180 after treatment (vs control p <0.0001). Similarly an increase in collagen deposition in the bladder wall was seen during the whole late radiation phase but with increasing values toward the end of the experiment (day 360 vs control p <0.0001). CONCLUSIONS: Irradiation of the bladder results in late damage to the barrier function of small subepithelial blood vessels, causing pronounced albumin leakage at the early steps of the chronic radiation reaction, clearly preceding excessive collagen deposition in the bladder wall. Current data suggest a vascular component in the development of late radiation induced changes in the bladder. PMID- 17085199 TI - Prognostic value of DNA ploidy, bcl-2 and p53 in localized prostate adenocarcinoma incidentally discovered at transurethral prostatectomy. AB - PURPOSE: Discovery of prostatic adenocarcinoma limited to transurethral resection material generates a treatment dilemma. We investigated the usefulness of parameters shown to be associated with prognosis in prostate cancer (p53 and bcl 2 immuno-expression, DNA cell cycle analysis and Gleason score) to stratify these incidentally identified tumors to guide clinical decision making. MATERIALS AND METHODS: Paraffin embedded tissues from transurethral prostate resection specimens containing T1a prostate adenocarcinoma from 44 patients who underwent resection between 1980 and 1990 were immunostained for p53 and bcl-2, and subjected to flow cytometry to determine DNA ploidy. Gleason score was determined by 2 pathologists independently. Statistical relationships among these 4 variables, tumor progression and cancer specific survival were analyzed. RESULTS: Six of 44 patients in the study population had cancer progression. Time to clinical progression was 4.5 years (range 7 months to 11 years). Most tumors stained negative for p53 and bcl-2. Only 2 tumors studied were aneuploid and neither of these 2 patients had cancer progression. Only Gleason score was a significant predictor of cancer progression on univariate and multivariate Cox regression analysis (p = 0.045 and 0.046, respectively). No tumor characteristics correlated with time to disease progression, including p53 and bcl-2 immuno expression, and Gleason score (p = 0.182, 0.563 and 0.346, respectively). Positive immunostaining for p53 and bcl-2 did not occur together in the same tumor in significant fashion (p = 0.334), nor did either significantly occur more with aneuploidy (p = 0.237 and 0.307 respectively). CONCLUSIONS: For T1a prostate cancer incidentally detected on transurethral prostate resection p53 and bcl-2 immuno-expression, and DNA ploidy do not predict survival or disease progression. PMID- 17085200 TI - Air pockets trapped during routine coupling in dry head lithotripsy can significantly decrease the delivery of shock wave energy. AB - PURPOSE: Current lithotriptors use a dry treatment head that must be coupled to the patient with gel or oil. We determined how the quality of coupling affects stone breakage under conditions that simulated patient treatment. MATERIALS AND METHODS: Experiments were performed with a Dornier (DoLi-50 electromagnetic lithotriptor. The test tank had a clear Mylar membrane for coupling with the treatment head water cushion. Thus, air pockets trapped at the coupling interface could be photographed for quantitation. Coupling efficiency was assessed using a fiberoptic hydrophone and different coupling regimes were tested for the effect on gypsum stone breakage. RESULTS: The quality of coupling was variable with air pockets covering 1.5% to 19% of the coupling area, resulting in a mean decrease in shock wave amplitude of approximately 20%. Breaking and reestablishing contact, as when a patient is repositioned during treatment, decreased acoustic pressure almost 32%, representing a 57% decrease in acoustic energy transmission. Stone breakage was also decreased when air was trapped in coupling and only 2% coverage by air pockets decreased stone breakage by 20% to 40%. CONCLUSIONS: These in vitro results suggest that coupling in lithotripsy can pose a significant barrier to the transmission of shock wave energy to the patient. Stone breakage was sensitive to air pockets at the coupling interface. Recoupling was particularly disruptive, suggesting that repositioning the patient could substantially degrade coupling quality. It seems reasonable that variability in the quality of coupling could contribute to variability in clinical outcomes. PMID- 17085201 TI - Functional analysis of active urethral closure mechanisms under sneeze induced stress condition in a rat model of birth trauma. AB - PURPOSE: We evaluated changes in the urethral closure mechanism under a sneeze induced stress condition in a rat model of birth trauma. MATERIALS AND METHODS: Four days after vaginal distention induced by balloon catheter inflation in the vagina sneezing was induced while recording intravesical pressure with the rat under urethane anesthesia to evaluate sneeze induced leak point pressure, defined as the lowest pressure inducing fluid leakage from the urethral meatus during sneezing. Sneeze induced responses in the bladder and proximal or mid urethra were also measured using microtip transducer catheters. RESULTS: In 5 sham operated rats no leakage was observed from the urethral meatus during sneezing, which produced an increase in intravesical pressure of up to 34 cm H(2)O. However, in 5 of 6 rats with vaginal distention leakage during sneezing was observed with a sneeze leak point pressure of 26.2 cm H(2)O. In the mid urethra microtip transducer catheters revealed that pressure increases during sneezing were greater than those in the bladder but they were significantly decreased in the 5 incontinent vaginal distention rats. However, sneeze induced responses at the proximal urethra, which were similar to those in the bladder, were not different in sham operated and incontinent vaginal distention rats. CONCLUSIONS: Sneeze induced stress urinary incontinence in a rat model of birth trauma was caused by decreased active closure mechanisms at the mid urethra without affecting the passive transmission of abdominal pressure in the proximal urethra. PMID- 17085203 TI - Detrusor quantitative morphometry in obstructed males and controls. AB - PURPOSE: We studied the usefulness of computer assisted morphometry for measuring detrusor muscle cell diameter and the connective tissue-to-smooth muscle ratio in patients with bladder outlet obstruction, acute urinary retention and a nonobstructed control group. MATERIALS AND METHODS: A prospective study was done in patients with bladder outlet obstruction undergoing transurethral prostate resection. Patients were divided into 33 with obstruction and 14 in acute urinary retention. A total of 15 males without obstruction undergoing transurethral prostate resection for bladder tumor formed the control group. Detrusor specimens were obtained during transurethral prostate resection. Detrusor muscle cell diameter was measured using light microscopy and a semiautomatic image analysis system. The connective tissue-to-smooth muscle ratio was automatically determined with computer assisted image analysis. Symptoms and urodynamic assessment were performed preoperatively and 6 months postoperatively. RESULTS: A total of 62 patients were included. The obstruction and acute urinary retention groups had a statistically higher detrusor muscle cell diameter and more fibrosis than the control group. Patients in acute urinary retention had more intrafascicular fibrosis (higher connective tissue-to-smooth muscle ratio at 40x magnification) than patients with obstruction. There were no differences in detrusor muscle cell diameter or interfascicular fibrosis (connective tissue-to-smooth muscle ratio at 10x magnification) between the obstruction and acute urinary retention groups. Detrusor muscle cell diameter correlated with symptom duration and functional recovery after transurethral prostate resection. Detrusor fibrosis correlated with preoperative detrusor pressure at maximum flow and postoperative compliance. Patients in acute urinary retention had fewer symptoms and higher residual volume. Other urodynamic parameters and their improvement after surgery were similar in the acute urinary retention and obstruction groups. CONCLUSIONS: Morphometric differences in detrusor muscle cell diameter and the connective tissue-to-smooth muscle ratio were observed between controls and patients with obstruction. There is an increase in detrusor muscle cell diameter and fibrosis in bladder outlet obstruction and more intense intrafascicular collagen deposition in patients in acute urinary retention. PMID- 17085202 TI - Partial bladder outlet obstruction selectively abolishes protein kinase C induced contraction of rabbit detrusor smooth muscle. AB - PURPOSE: Despite the acute onset, partial bladder outlet obstruction in the rabbit induces detrusor remodeling similar to that in men with benign prostatic hyperplasia in terms of its impact on structural and functional alterations in smooth muscle. We determined if partial bladder outlet obstruction induced remodeling alters the protein kinase C signaling pathway that leads to contraction. MATERIALS AND METHODS: Smooth muscle from control animals and those subjected to 2 weeks of partial bladder outlet obstruction were mounted for isometric force recording, measurement of myosin light chain phosphorylation and levels of adducin phosphorylation. Bladder muscle strips were stimulated by phorbol dibutyrate or carbachol in the presence and absence of bisindolylmaleimide-1. RESULTS: Smooth muscle strips from animals subjected to partial bladder outlet obstruction showed little to no increase in stress in response to phorbol dibutyrate and no increase in myosin light chain phosphorylation levels. Muscle strips from control animals produced a robust contraction with concomitant increases in myosin light chain phosphorylation. Inhibition of protein kinase C by bisindolylmaleimide-1 significantly depressed carbachol induced contractions of muscle strips from control animals but it had no effect on carbachol induced contractions of muscle strips from outlet obstructed animals. Phorbol dibutyrate increased phospho-adducin levels in muscle strips from the 2 animal sources, suggesting that protein kinase C could be activated. CONCLUSIONS: We propose that partial bladder outlet obstruction does not alter protein kinase C activation, but rather abolishes or uncouples the pathway(s) downstream of protein kinase C, leading to contraction. Loss of this pathway may contribute to the loss of normal voiding behavior and the resultant decompensated state. PMID- 17085204 TI - The striated urethral sphincter of the pig shows morphological and functional characteristics essential for the evaluation of treatments for sphincter insufficiency. AB - PURPOSE: New treatments are currently under investigation for intrinsic sphincter insufficiency. However, animal models in which to study the resting urethral tone generated by the striated urethral sphincter are still lacking. We describe the striated urethral sphincter in the pig. We investigated its participation in resting urethral tone with the aim of developing new tools for evaluating therapies for sphincter insufficiency. MATERIALS AND METHODS: A total of 15 female pigs were used in this study. Anatomy of the striated urethral sphincter was described via transpubic and endoscopic approaches. Participation of the striated urethral sphincter in resting urethral tone was assessed by analysis of urethral pressure profilometry (maximum urethral closure pressure, functional urethral length and area under the curve) before and after curare injection or by destruction of the striated urethral sphincter by endoscopic electrocautery. Serial urethral cross sections were immunostained for slow/fast myosin and digitalized for 3-dimensional reconstructions to determine striated urethral sphincter volume. RESULTS: The striated urethral sphincter was Omega-shaped and it encircled the distal third of the urethra. A mean peak intraurethral pressure +/- SEM of 58.9 +/- 13.4 cm H(2)O was noted in front of the striated urethral sphincter. Curare injection and endoscopic injury decreased maximum urethral closure pressure by 48.4% and 51.1%, functional urethral length by 10.3% and 15.3%, and area under the curve by 47% and 64%, respectively. The striated urethral sphincter consisted of 52% slow and 48% fast myofibers. Its mean volume was 0.87 cm. CONCLUSIONS: The striated urethral sphincter of the female pig shows the morphological and functional features of a tonic muscle. Methods of measuring resting urethral tone generated by the striated urethral sphincter represent original tools for evaluating therapies for intrinsic sphincter insufficiency. PMID- 17085205 TI - A P300 event related potential technique for assessment of sexually oriented interest. AB - PURPOSE: Despite all of the modern, sophisticated tests that exist for diagnosing and assessing male and female sexual disorders, to our knowledge there is no objective psychophysiological test to evaluate sexual arousal and interest. We provide preliminary data showing a decrease in auditory P300 wave amplitude during exposure to sexually explicit video clips and a significant correlation between the auditory P300 amplitude decrease and self-reported scores of sexual arousal and interest in the clips. MATERIALS AND METHODS: A total of 30 healthy subjects were exposed to several blocks of auditory stimuli administered using an oddball paradigm. Baseline auditory P300 amplitudes were obtained and auditory stimuli were then delivered while viewing visual clips with 3 types of content, including sport, scenery and sex. RESULTS: Auditory P300 amplitude significantly decreased during viewing clips of all contents. Viewing sexual content clips caused a maximal decrease in P300 amplitude (p <0.0001). In addition, a high correlation was found between the amplitude decrease and scores on the sexual arousal questionnaire regarding the viewed clips (r = 0.61, p <0.001). In addition, the P300 amplitude decrease was significantly related to the sexual interest score (r = 0.37, p = 0.042) but not to interest in clips of nonsexual content. CONCLUSIONS: The change in auditory P300 amplitude during exposure to visual stimuli with sexual context seems to be an objective measure of subject sexual interest. This method might be applied to assess therapeutic intervention and as a diagnostic tool for assessing disorders of impaired libido or psychogenic sexual dysfunction. PMID- 17085206 TI - Re: The Prostate Cancer Prevention Trial: design, biases and interpretation of study results. P. J. Goodman, I. M. Thompson, Jr., C. M. Tangen, J. J. Crowley, L. G. Ford and C. A. Coltman, Jr., J Urol, 175: 2234-2242, 2006. PMID- 17085207 TI - Re: Cystectomy delay more than 3 months from initial bladder cancer diagnosis results in decreased disease specific and overall survival. C. T. Lee, R. Madii, S. Daignault, R. L. Dunn, Y. Zhang, J. E. Montie and D. P. Wood, Jr., J Urol, 175: 1262-1267, 2006. PMID- 17085209 TI - Re: Diagnostic value of systematic biopsy methods in the investigation of prostate cancer: a systematic review. K. Eichler, S. Hempel, J. Wilby, L. Myers, L. M. Bachmann and J. Kleijnen, J Urol, 175: 1605-1612, 2006. PMID- 17085210 TI - Re: Single center experience with endoscopic management of vesicoureteral reflux in children. J. C. Routh, D. R. Vandersteen, H. Pfefferle, J. J. Wolpert and Y. Reinberg, J Urol, 175: 1889-1893, 2006. PMID- 17085212 TI - Re: Low age adjusted free testosterone levels correlate with poorly differentiated prostate cancer. I. F. San Francisco, M. M. Regan, W. C. DeWolf and A. F. Olumi, J Urol, 175: 1341-1346, 2006. PMID- 17085214 TI - Re: The impact of late presentation of posterior urethral valves on bladder and renal function. O. Ziylan, T. Oktar, H. Ander, E. Korgali, H. Rodoplu and T. Kocak, J Urol, 175: 1894-1897, 2006. PMID- 17085216 TI - Re: A new classification is needed for pelvic pain syndromes--are existing terminologies of spurious diagnostic authority bad for patients? P. Abrams, A. Baranowski, R. E. Berger, M. Fall, P. Hanno and U. Wesselmann, J Urol, 175: 1989 1990, 2006. PMID- 17085217 TI - Language processing across the life span: new methodologies to study old questions. PMID- 17085219 TI - Ethics, legal issues, and consent for fillers. AB - The huge patient demand for filler rejuvenation exposes plastic surgeons to legal issues. Plastic surgery statistics confirm a trend toward nonsurgical procedures, with a slight increase in surgical procedures overall from 2004 to 2005. Nonsurgical procedures, especially fillers, exhibited a 35% increase. Patients are seeding a quick-recovery, minimally invasive procedure that provides them maximal rejuvenation. This demand and media-driven exposure may create concerns for plastic surgeons as to patient selection and ethical, regulatory, and legal issues relating to filler choices and their use. This article addresses these concerns. PMID- 17085220 TI - Calcium hydroxylapatite facial filler (Radiesse): indications, technique, and results. AB - This article discusses the use of Radiesse; a semisolid, cohesive subdermal injectable implant. Through direct and invaluable experience gained over time, plastic surgeons will decide which areas of the face may best be suited for Radiesse and which may be appropriate for other fillers. PMID- 17085221 TI - Hyaluronic acids. AB - Hyaluronic acids (HAs) are the most versatile fillers on the market. Since their approval in Canada in 1998 and the United States in 2003, applications for facial contouring have been varied, with predictable and outstanding results. Numerous publications support their safety, durability, longevity, and application to reconstructive and cosmetic procedures for both surgical and nonsurgical applications. The incidence of complications is low, and the treatment of complications is relatively straightforward with the use of hyaluronidase. This article discusses the uses of the HAs in general, with regard to the various particles sizes (Restylane, Restylane Fine Lines, Perlane, Juvederm 18, 24, 30), which are available in Canada and will be available in the United States in the fall of 2006. PMID- 17085222 TI - Sculptra: the new three-dimensional filler. AB - Sculptra, the synthetic injectable poly-l-lactic acid (PLLA), is a revolutionary three-dimensional filler lasting 18 to 24 months. This unique volumizing agent is best used to globally restore volume to the lower two thirds of the face in patients who have lipoatrophy. Sculptra is a biocompatible, biodegradable, and nonimmunogenic derivative of the alpha-hydroxy-acid family. The size and the slow degradation kinetics of PLLA microparticles act as a stimulus for collagen production, providing lasting volume enhancement in lipoatrophy patients. PMID- 17085223 TI - ArteFill: a third-generation permanent dermal filler and tissue stimulator. AB - The technical and physiologic shortcomings associated with prior generations of polymethylmethacrylate (PMMA) dermal fillers have been reduced significantly since their original market introduction in 1989. For example, the PMMA microspheres in the third-generation product ArteFill meet the new limit set by the FDA of less than 1 particle below 20 micrometers per 100 microspheres (< 1% by the number). Therefore, it is expected that the already low granuloma rate following injections with second-generation product Artecoll (1:5000) will be reduced further with injections of ArteFill. In the authors' experience, both Artecoll and ArteFill have proven to be safe, predictable, and effective dermal fillers. It is anticipated that Artefill will receive FDA approval in late 2006 and will be marketed internationally. PMID- 17085224 TI - Facial augmentation with structural fat grafting. AB - Fat grafting through a blunt cannula has been used by plastic surgeons for altering facial contours for 100 years. Autologous tissue is completely biocompatible and is usually the safest choice for altering facial volume or contours. Fat grafts can be placed in such a fashion that they are long lasting, completely integrated, and natural appearing. Only in the past 20 years have advances in techniques and instrumentation allowed us to obtain predictable results that make fat grafting a viable option for soft tissue augmentation. Our understanding of aging and methods of rejuvenation have developed also. We now approach rejuvenation and adjustment of facial proportion with a better understanding of the need for the restoration or adjustment of facial volume. PMID- 17085225 TI - Vascularized fat transfer. AB - The use of free tissue transfers has dramatically improved the outcomes of soft tissue reconstructions of the face. This is particularly true when large volumes of soft tissue are needed or when the recipient bed has a compromised local blood supply. Although traditional fat grafting procedures often allow for satisfactory results in some patients, the use of scapular and parascapular free tissue transfers has markedly improved the predictability of these reconstructions, often in a single stage. As perforator flap reconstructions evolve over time, these types of flaps may eventually supplant the available fasciocutaneous free flaps as the flap of choice in these reconstructions primarily because of improved donor site scarring and morbidity. PMID- 17085226 TI - Collagen-containing fillers: alone and in combination. AB - Soft tissue fillers may be injected into facial skin to ameliorate the appearance of fine lines and wrinkles, to augment lips and hollow cheeks, and to correct other imperfections, ultimately producing a more youthful appearance. Animal- and human-derived collagen implants have been used successfully for soft tissue augmentation for several years. Bovine collagen fillers were the most popular injectable implants for nearly 2 decades in the United States. Although these are still used, human bioengineered collagen products and fillers that contain hyaluronic acid have become available and are now more commonly used than the bovine-derived products. This article discusses the rationale for using the various types of fillers in combination and describes techniques for combining these fillers. PMID- 17085227 TI - Legal ramifications of off-label filler use. AB - Dermal fillers represent one of the most commonly used noninvasive cosmetic treatments. They are often used in an off-label manner. Most off-label use is not only legal, but represents an appropriate physician standard of care. This article will first describe the meaning of on- and off-label. Then the article will explore manufacturer promotion of off-label use of both drugs and devices. Finally, the legal ramifications of off-label dermal filler use will be discussed. PMID- 17085229 TI - Antioxidants in veterinary nutrition. AB - Nutritional antioxidants have experienced a surge in research and interest in the past 20 years. this surge may be attributed to the improved methodology for investigation as well as the focus on diseases and aging processes related to oxidative stress that lend themselves to opportunistic outcomes. As such, the field of veterinary nutritional antioxidant research is also beginning to yield some interesting results, albeit, small in number compared with laboratory animals and human beings. Nonetheless, this article updates the practitioner on recent advances in research involving nutritional antioxidant applications in companion animals. PMID- 17085230 TI - Nutrition and immune function. AB - The complexity of the immune system allows for a multitude of potential avenues for nutrient modulation, but this also increase the challenge of producing a predictable in vivo response. Because the immune response is a cascade of biologic events, development of nutritional support paradigms cannot and should not be made in a vacuum or with the expectation of a singular response. It is absolutely imperative that the clinician/nutritionist understand the differences in metabolIc and physiologic responses to disease states (ie, shock, trauma, organ-specific dysfunction) so as to maximize immunocompetence through specialized feeding practices. This level of understanding is invaluable, especially when considering the possible benefit of nutrient combinations for immunomodualtion. PMID- 17085231 TI - Nutrition in critical illness. AB - Malnutrition associated with critical illness has been unequivocally associated with increased morbidity and mortality in humans. Because malnutrition may similarly affect veterinary patients, the nutritional requirements of hospitalized critically ill animals must be properly addressed. Proper nutritional support is increasingly being recognized as an important therapeutic intervention in the care of critically ill patients. The current focus of veterinary critical care nutrition, and the major focus of this article, is on carefully selecting the patients most likely to benefit from nutritional support, deciding when to intervene, and optimizing nutritional support to individual patients. PMID- 17085232 TI - Management of anorexia in dogs and cats. AB - The management of anorexia should center first on the urgent and emergent medical management of the patient and be followed by feeding of a highly palatable food in a low-stress environment and manner. Diet palatability can potentially be improved by increasing dietary moisture, fat, or protein, and, in the dog, by adding sugar or salt as well as by using a variety of fresh, pleasantly aromatic, and uncommon foods. Caution should be used when increasing or adding nutrients that may be harmful to patients with specific diseases. Concurrent drug therapy that may reduce appetite should be minimized, and physical barriers to eating should be removed. Patients that consume less than resting energy requirement of longer than 3 to 5 days with no trend toward improving should receive parenteral or enteral nutrition. PMID- 17085233 TI - Hydrolyzed protein diets for dogs and cats. AB - The primary aim of a hydrolyzed protein diet is to disrupt the proteins within the diet sufficiently to remove existing allergens. Published assessment of hydrolyzed protein diets includes physiochemical and immunologic assays as well as nutritional and clinical feeding trials. Potential problems include poor palatability, hypoosmotic diarrhea, and a reduced nutritional value, although persistent allergenicity is the most significant. The primary indications for a hydrolyzed protein diet are use in elimination trials for the diagnosis of adverse food reactions, and the initial management of inflammatory bowel disease. Initial studies of hydrolyzed diet efficacy are encouraging. Consideration of the source ingredients should be given when using hydrolyzed protein diets in elimination feeding trials because antigenic sites may not be fully destroyed. PMID- 17085234 TI - Unconventional diets for dogs and cats. AB - Food plays a far more complex role in daily life than simply serving as sustenance. Social and cultural factors along with individual beliefs govern people's eating behaviors, and it is likely that these same factors influence their choice of diet and feeding practices for their pets. Some people seek alternatives to conventional commercial pet foods, including commercially available "natural" diets, raw food diets, and vegetarian diets, in addition to a variety of home-prepared diets. Exploring a person's knowledge and beliefs about feeding pets can aid in understanding her or his motives for seeking alternative and may help in changing those practices when it is in the best interest of the pet to do so. PMID- 17085235 TI - Understanding and managing obesity in dogs and cats. AB - Treatment of diabetes mellitus (DM) is a common disease, and nutrition has become an integral part of management. This article focuses on the role of nutrition and dietary ingredients in OA, evaluating current evidence for obesity management, omega-3 fatty acids, and chondromodulating agents. Additionally, Keeping an animal in optimal to slightly lean body condition has been shown to decrease the risk of development of OA and to aid in management of dogs with OA. PMID- 17085236 TI - Feline diabetes mellitus: low carbohydrates versus high fiber? AB - Treatment of diabetes mellitus (DM) in the cat relies primarily on the adequate insulin therapy and controlled dietary intake. The goals of managing DM in the cat have changed from attaining glycemic control to achieving diabetic remission (transient diabetes) in a large proportion of cases. Remission rates of up to 68% have been published. The used of low-carbohydrate foods for cats improves the odds of achieving diabetic remission by fourfold. Nonetheless, some cats show an improved response to high-fiber food. Clinical judgement, trial, and personal preference to currently dictate which diet to offer an individual animal. PMID- 17085237 TI - Nutrition and osteoarthritis in dogs: does it help? AB - Osteoarthritis (OA) is a common disease, and nutrition has become an integral part of management. This article focuses on the role and dietary ingredients in OA, evaluating current evidence for obesity management, omega-3 fatty acids, and chondromodulating agents. Additionally, keeping an animal in optimal to slightly lean body condition has been shown to decrease the risk of development of OA and to aid management of dogs with OA. PMID- 17085238 TI - Taurine and carnitine in canine cardiomyopathy. AB - Some newer more promising therapies for dogs with dilated cardiomyopathy (DCM) are taurine and carnitine. Deficiencies of these nutrients have been shown to cause DCM in dogs, and some breeds of dogs have shown dramatic improvement in myocardial function after supplementation with one or both nutrients. Although most dogs diagnosed with DCM do not have a documented taurine or carnitine deficiency, they may still be benefit from supplementation. These nutrients are safe to administer to dogs. For some owners, the high cost of carnitine is the only deterrent to giving their dogs supplements of both nutrients. PMID- 17085239 TI - Nutritional risks to large-breed dogs: from weaning to the geriatric years. AB - Distinctive risk factors for disease are identifiable throughout life stages of large- and giant-breed dogs. From weaning to maturity, improper nutrition is linked to developmental skeletal diseases. As large dogs mature, skeletal diseases and obesity can lead to osteoarthritis. These dogs are difficult to manage when orthopedic or osteoarthritic disease affects mobility and quality of life, thereby increasing the risk of early death. Gastric dilatation and volvulus is another disease that is leading cause of death in large- and giant-breed dogs. Management of health, including proper nutrition, exercise, and weight control, provides the best opportunity for successful aging of large- and giant-breed dogs. PMID- 17085240 TI - Nutrition and lower urinary tract disease in cats. AB - Lower urinary tract disease occurs commonly in cats and is often associated with crystal-related disease. Dietary modification is beneficial in managing some of these diseases, including idiopathic cystitis, urolithiasis, and urethral matrix crystalline plugs. Altering dietary formulation may result in decreasing urinary concentrations of crystallogenic compounds, increasing urinary concentrations fo crystallogenic inhibitors, and diluting urine composition. PMID- 17085241 TI - Nutritional management of chronic renal disease in dogs and cats. AB - Chronic renal disease is a leading cause of death in dogs and cats. Recent clinical studies show that nutrition plays a key role in improving quality of life and life expectancy of these patients. Typical nutritional interventions include modifying the protein, phosphorus, and lipid concentrations. Nutritional therapy, however, does not simply mean changing the diet; consideration must also be given to ensuring adequate caloric intake and to the method of feeding. Monitoring the effects of the dietary therapy is also crucial to ensure that the patients are responding appropriately to the selected nutritional modifications. Nutritional management must be coordinated with medical management for long term successful treatment. PMID- 17085242 TI - Dietary influences on periodontal health in dogs and cats. AB - A pet cannot be healthy without oral health. Periodontal is a significant disease that has local and systemic ramifications. It has been stated earlier that effective plaque control prevents gingivitis. In human beings, 90% of periodontitis occurs as the result of progression gingivitis, and this type of periodontitis can be completely prevented by plaque control. It is reasonable that dogs and cats react similarly and that effective plaque control could prevent a large percentage of periodontitis cases. Proper nutrition and effective oral hygiene are necessary components of oral health and should be jointly promoted in the management of oral disease in dogs and cats. PMID- 17085244 TI - Clinical epidemiology of acute lung injury and acute respiratory distress syndrome: incidence, diagnosis, and outcomes. AB - Acute respiratory distress syndrome (ARDS) and acute lung injury (ALI) are clinical syndromes characterized by the sudden onset of severe hypoxemia and diffuse bilateral pulmonary infiltrates in the absence of left atrial hypertension. Although advances have been made in the understanding of the etiology, pathophysiology, and epidemiology of both entities, many questions remain regarding their incidence, diagnosis, and outcomes. This article reviews the currently used definition of ARDS and ALI, different studies that have advanced the understanding of the epidemiology and outcomes of these entities, and several diagnostic issues that are important for both clinicians and researchers. PMID- 17085245 TI - The role of CT-scan studies for the diagnosis and therapy of acute respiratory distress syndrome. AB - CT has provided new insights on the pathophysiology of acute respiratory distress syndrome (ARDS), demonstrating that ARDS does not affect the lung parenchyma homogeneously. These findings suggest that lung edema, as assessed by CT scan, should be included in the definition. Lung CT findings may provide a firm rationale for tailoring tidal volume during mechanical ventilation. Ideally, tidal volume should be proportional to the portion of the lung open to ventilation, as assessed by CT scan, rather than to the body weight. CT assessment of lung recruitability seems to be a prerequisite for a rational setting of positive end-expiratory pressure. PMID- 17085246 TI - Pathology of acute lung injury and acute respiratory distress syndrome: a clinical-pathological correlation. AB - Clinical criteria cannot identify all patients who develop acute respiratory distress syndrome, which might indicate the need to develop new tools that improve reliability and diagnostic accuracy. Nevertheless, being conscious of the limitations of the American-European consensus conference criteria continues to be useful in clinical daily practice because these patients would benefit from treatments. This and other management strategies have been evaluated in patients who, fulfilling the clinical criteria of acute respiratory distress syndrome, could not have diffuse alveolar damage. PMID- 17085247 TI - Mechanisms of acute lung injury/acute respiratory distress syndrome. AB - Acute lung injury/acute respiratory distress syndrome is a clinical syndrome that describes a single common end point: severe injury to the alveolar capillary membrane and the development of proteinaceous edema. Forty years of clinical and basic science research have elaborated many of the pathophysiologic mechanisms that link initiating insults to the development of alveolar membrane failure. A combination of genetics, comorbid conditions, inciting events, and factors yet to be discovered may uniquely alter the expression of these mechanisms in the individual patient. PMID- 17085248 TI - Newly recognized causes of acute lung injury: transfusion of blood products, severe acute respiratory syndrome, and avian influenza. AB - Acute lung injury/acute respiratory distress syndrome (ALI/ARDS) is a clinical syndrome that has an ever-growing list of potential causes. The transfusion of blood products is often a life-saving therapy, but it can be associated with the development of ALI/ARDS. Transfusion-related ALI is now the leading cause of transfusion-associated fatalities in the United States. Two infectious causes of ALI/ARDS, severe acute respiratory syndrome and H5N1 influenza, have recently emerged and have the potential for pandemic spread. This article discusses the clinical importance, pathogenesis, diagnosis, management, and prevention of these newly recognized causes of respiratory failure. PMID- 17085249 TI - Conventional mechanical ventilation in acute lung injury and acute respiratory distress syndrome. AB - Acute lung injury and acute respiratory distress syndrome are inflammatory conditions involving a broad spectrum of lung injury from mild respiratory abnormality to severe respiratory derangement. Regardless of cause (direct or indirect lung injury), pulmonary physiology and mechanics are altered, leading to hypoxemic respiratory failure. the use of positive pressure ventilation itself may cause lung injury (ventilator-induced lung injury, or VILI). VILI may amplify preexisting injury, delay lung recovery, and result in adverse outcomes. This article examines the evidence supporting lung-protective ventilation strategies and addresses the methods, outcomes, and potential obstacles to implementation of such approaches. PMID- 17085250 TI - New modalities of mechanical ventilation: high-frequency oscillatory ventilation and airway pressure release ventilation. AB - Management of acute lung injury (ALI) and the acute respiratory distress syndrome (ARDS) is largely supportive, with the use of mechanical ventilation being a central feature. Recent advances in the understanding of ALI/ARDS and mechanical ventilation have revealed that lung-protective ventilation strategies may attenuate ventilator-associated lung injury and improve patient morbidity/mortality. High-frequency oscillatory ventilation and airway pressure release ventilation are two novel alternative modes of ventilation that theoretically fulfill the principles of lung protection and may offer an advantage over conventional ventilation for ALI/ARDS. PMID- 17085251 TI - Pulmonary artery catheter and fluid management in acute lung injury and the acute respiratory distress syndrome. AB - The PAC provides a wealth of information about circulatory and respiratory systems and intravascular fluid volume over time. Specifically, the PAC allows measurement of central venous and pulmonary arterial pressure, pulmonary artery occlusion pressure, mixed venous blood gases, and indicator-dilution cardiac output. Based on these quantitative date, systemic and pulmonary vascular resistance can be derived. The PAC is frequently used in patients with ALI and ARDS, both to confirm the diagnosis and to optimize hemodynamic management. In this article, we review the evidence on the use of the PAC in patients with ALI/ARDS, paying particular attention to the recently published fluid and catheter treatment trial by the ARDS Clinical Trials Network. PMID- 17085252 TI - Adjunctive therapy to mechanical ventilation: surfactant therapy, liquid ventilation, and prone position. AB - Acute lung injury and acute respiratory distress syndrome are associated with significant morbidity and mortality in critically ill patients. Although lung protective mechanical ventilation is the only therapy shown to reduce mortality and development of organ failure, several biologic pathways have been identified and provided an opportunity for therapeutic interventions. No pharmacologic or adjunctive treatments are available. Clinical studies demonstrated that prone position results in significant and clinically relevant improvement in oxygenation and ventilation, which persist when patients are returned to supine position; the beneficial response is not limited to patients turned early in disease course. Few complications are associated with prone ventilation. Clinical experience suggests that prone ventilation may protect the lung from potential detrimental effects of mechanical ventilation. Further studies are needed. PMID- 17085253 TI - The alveolar-epithelial barrier: a target for potential therapy. AB - During acute lung injury (ALI), the alveolar-capillary barrier is damaged, resulting in the accumulation of fluid and protein in the alveolar space characteristic of the acute respiratory distress syndrome (ARDS). Disordered epithelial repair may contribute to the development of fibrosis and worsen outcomes in patients who have lung injury. This article discusses novel emerging therapies based on these mechanisms that are designed to preserve the function and promote the repair of the alveolar epithelium in patients who have ALI/ARDS. PMID- 17085254 TI - Therapy for late-phase acute respiratory distress syndrome. AB - Prolonged hypoxemic respiratory failure and evidence of lung organization and fibrosis are features of an ARDS subgroup that is variably identified as "late," "persistent," or "fibroproliferative" ARDS. Early reports suggested that patients with late ARDS had a high mortality unless treated with corticosteroids. A large recent study with improved methodology has demonstrated that despite improvements of pulmonary physiology, corticosteroids do not change mortality of patients who continue to meet ARDS criteria 7 to 28 days after onset of acute lung injury. Additionally, there is no compelling evidence that persistent ARDS confers a higher mortality than that of ALI/ARDS. Observational and interventional studies are needed to increase understanding of the incidence, best management, and outcomes of patients with persistent ARDS. PMID- 17085255 TI - Quality of life, emotional abnormalities, and cognitive dysfunction in survivors of acute lung injury/acute respiratory distress syndrome. AB - This article provides a brief discussion of the recent literature linking quality of-life impairment to physical dysfunction after acute lung injury/acute respiratory distress syndrome. Its main focus is a review of the current knowledge concerning cognitive and emotional outcomes after lung injury and their impact on long-term quality of life. PMID- 17085256 TI - Neuromuscular sequelae in survivors of acute lung injury. AB - Pulmonary function improves rapidly in most survivors of ALI, yet most have profound and life-altering physical impairment. critical illness polyneuropathy and myopathy (CIPM) are complications that may account for most long-term disability. Short-term morbidity includes difficulty weaning from mechanical ventilation, return to assisted breathing after weaning, and delayed return home after hospital discharge. Therapies that reduce sepsis and prolonged mechanical ventilation may minimize CIPM, as in intensive insulin treatment of ICU hyperglycemia. CIPM identification is important for ALI patients' care and prognostication. Observational studies are needed to understand the epidemiology and natural history of CIPM; interventional studies with CIPM and functional outcomes as major endpoints are essential for improvement of ALI patients' health and quality of life. PMID- 17085257 TI - Genetic epidemiology of acute respiratory distress syndrome: implications for future prevention and treatment. AB - The genetic susceptibility to the development of and variable outcomes in acute lung injury/acute respiratory distress syndrome (ALI/ARDS) has become a topic of great interest in the pulmonary and critical care community. Published studies of variable genetic susceptibility to ALI/ARDS already have identified some important candidate genes and potential gene-environment interactions. This article reviews these recent studies, features of the current approach, and implications for future prevention and treatment in ALI. The challenges and potential contributions of genetic epidemiology to the future prevention and treatment in ALI are discussed. PMID- 17085258 TI - Why do patients who have acute lung injury/acute respiratory distress syndrome die from multiple organ dysfunction syndrome? Implications for management. AB - Acute respiratory distress syndrome (ARDS) affects some 10% to 15% of ICU patients and is associated with mortality rates of 40% to 50%. Although ARDS is the most severe form of acute respiratory failure, refractory hypoxia is an uncommon cause of death in these patients. The majority of patients who have ARDS die from multiple-organ dysfunction syndrome (MODS), and ARDS should, therefore, be seen as a systemic disease. Improved understanding of the systemic factors involved in the development and evolution of ARDS and MODS should facilitate the development of new therapeutic agents that will improve outcomes in these patients. PMID- 17085259 TI - Acute lung injury and the acute respiratory distress syndrome: challenges in clinical trial design. AB - Despite significant advances in the understanding of the complex pathophysiology, only a limited number of new treatments for acute lung injury (ALI) have emerged in the last 2 decades. This article discusses some of the challenges that remain in conducting clinical research in patients who have ALI and acute respiratory distress syndrome. New definitions that incorporate prognostic measures and reduce patient heterogeneity will allow more efficient enrollment of patients. Delineating outcomes attributable to the lung injury will improve the power of studies to detect significant treatment effects. Future collaborative studies will be needed to investigate longer-term clinical outcomes. PMID- 17085260 TI - Spiritual healing: more than meets the eye. PMID- 17085261 TI - Exacerbation of paraneoplastic syndrome of inappropriate antidiuretic hormone by parenteral nutrition in a patient affected by a large-cell neuroendocrine pancreatic cancer. PMID- 17085262 TI - Efficacy of pregabalin in the management of cetuximab-related itch. PMID- 17085263 TI - A case of visceral post-stroke pain. PMID- 17085264 TI - Central post-stroke syndrome treated with parenteral lidocaine. PMID- 17085265 TI - Topiramate resolves headache from pseudotumor cerebri. PMID- 17085266 TI - Symptom experience in the last year of life among individuals with cancer. AB - Individuals with cancer often experience many symptoms that impair their quality of life at the end of life. This study examines symptom experience at end of life among individuals with cancer, and determines if symptom experience changes with proximity to death, or differs by depressive symptomatology, sex, site of cancer, or age. A secondary analysis of data from three prospective, descriptive, longitudinal studies (n=174) was performed, using a three-level hierarchical linear model. Fatigue, weakness, pain, shortness of breath, and cough were the five most prevalent symptoms in the last year of life. The symptom experience in the last year of life was significantly associated with site of cancer, depressive symptomatology, dependencies in activities of daily living, and independent activities of daily living at the start of the study. These findings shed light on the symptom experience in the last year of life for individuals with cancer. With greater understanding of the symptom experience, intervention strategies can be targeted to achieve the desired outcome of increased quality of life at the end of life. PMID- 17085267 TI - Opioid escalation in patients with cancer pain: the effect of age. AB - Elderly people are commonly considered more susceptible to opioid effects. However, no data regarding the need for opioid escalation in patients already receiving opioids for the management of chronic pain are available. The purpose of this study was to evaluate the differences between younger and older patients during the crucial phase of opioid titration. One hundred consecutive patients with cancer pain requiring further opioid dose refinement were recruited for this cohort study. Pain intensity, dose of opioids, number of opioids used (need to switch), routes of administration used, and opioid-related symptoms were measured from admission until dose stabilization. Opioid escalation indexes (OEIs) were calculated. For the purpose of analysis, patients were divided into three age groups (<65, 65-74, 75 or over). Despite differences in opioid doses at admission (lower in older patients), no differences were found in routes, need to switch, OEI, or other parameters between younger and older patients. Similarly, adverse effects did not significantly differ between the three groups, although an overall distress score worsened in older patients during acute titration and then improved at stabilization time. These data contradict the assumption that older patients who already receive opioids are more susceptible than younger adults to opioid effects during opioid titration. Although the elderly require lower doses, opioid effects do not appear to vary with age in this population. However, the group of patients over 75 was relatively small and data should be interpreted with caution. Careful titration based on the individual response seems appropriate irrespective of age. PMID- 17085268 TI - Measuring fatigue in Parkinson's disease: a psychometric study of two brief generic fatigue questionnaires. AB - This study evaluated and compared the measurement properties of the 13-item Functional Assessment of Chronic Illness Therapy-Fatigue Scale (FACIT-F) and the 9-item Fatigue Severity Scale (FSS) in 118 consecutive Parkinson's disease (PD) patients, using traditional and Rasch measurement methodologies. Both questionnaires exhibited excellent data quality and reliability (coefficient alpha>or=0.9), and acceptable rating scale functionality, and both discriminated between fatigued and nonfatigued patients. Factor and Rasch analyses provided general support for unidimensionality of both FACIT-F and FSS, although they do not appear to measure identical aspects of fatigue. No signs of differential item functioning (DIF) were found for the FACIT-F, whereas potential age DIF was detected for two FSS items. These results support the measurement validity of both questionnaires in PD, although the FACIT-F displayed better measurement precision and modest psychometric advantages over the FSS. Availability of psychometrically sound fatigue measures that are applicable across disorders provides a sound basis for advancing the understanding of this common and distressing complaint. PMID- 17085269 TI - The impact of providing symptom management assistance on caregiver reaction: results of a randomized trial. AB - This research evaluates the impact of a randomized clinical trial for lowering family members' reactions and increasing their involvement in assisting cancer patients undergoing chemotherapy to better manage their symptoms. The cognitive behavioral intervention was directed toward both the patient and family caregivers. At 10 weeks, caregivers who received the experimental intervention had significantly lower total reactions to assisting with symptoms and assisted with significantly fewer symptoms (linked to patient improvement in symptom severity) than caregivers receiving conventional care alone. Female caregivers receiving the intervention reported significantly lower reaction per symptom where assistance was provided compared to female caregivers in the control group. Thus, the intervention did not increase caregiver involvement in symptom management, quite possibly due to reduced symptom severity among patients in the intervention group. This research offers a model for evaluating trials directed toward both members of a dyad. PMID- 17085270 TI - Multidisciplinary management of cancer pain: a longitudinal retrospective study on a cohort of end-stage cancer patients. AB - The epidemiology of cancer pain and the outcomes associated with pain treatments were investigated through a retrospective survey of 772 patients with advanced cancer. The cumulative prevalence of pain was 87%, including all types of cancer. Mean duration of pain was 6.9+/-8.1 months. The prevalence of pain was 28%, 46%, 67%, 75%, and 79% at 6 months, 3 months, 1 month, 1 week, and 1 day before the time of death, respectively. The so-called "strong" opioids had been used in 85% of the 669 patients with pain. Seventy-nine percent of patients with pain received nonsurgical antineoplastic treatment for pain control. No more than 11% of patients ultimately experienced substantial pain in the last 6 months of life (defined as pain score 5-10 on a 0-10 numeric rating scale). We conclude that the application of a multidisciplinary approach to pain management offers effective pain control for most patients with advanced cancer. PMID- 17085271 TI - The utility of videoconferencing to provide innovative delivery of psychological treatment for rural cancer patients: results of a pilot study. AB - The unmet psychological needs of rural cancer patients are numerous. Telepsychology is a novel and feasible option that may provide cost-savings and help overcome inequalities in access to specialists. This is the first known study of psychological treatment for people with cancer delivered entirely via videoconferencing. We hypothesized that a telepsychology service would improve rural cancer patients' anxiety and depression levels and quality of life, and would be an acceptable, satisfactory, and practical mode of service delivery. Twenty-five cancer patients attended an average of three sessions with a clinical psychologist providing brief cognitive-behavioral therapy. Questionnaires were completed at pre-, post-, and 1-month follow-up. Patients benefited in terms of anxiety (P=0.01) and quality of life (P=0.04). The service was both practical and acceptable. These preliminary positive results provide a firm basis to conduct a randomized controlled trial of face-to-face interaction vs. videoconferencing. PMID- 17085272 TI - A survey of pain in idiopathic Parkinson's disease. AB - The aim of this study was to use both quantitative and qualitative data to assess pain in idiopathic Parkinson's disease (IPD) and classify it according to cause. In this descriptive cross-sectional study, 123 patients were interviewed and pain was assessed using patient descriptions, the Brief Pain Inventory, and Visual Analogue Scales. Patients reported 285 pains (median 2 per patient), and 22.8% had 4 or more pains. Pain was reported as a problem in 85% and was IPD-related in 62.6% of patients, unrelated to IPD in 64.2%, indirectly related to IPD in 8.1%, related to multiple causes in 4.1%, and treatment related in 0.8%. Pain unrelated to IPD was more common, more constant, and more severe than IPD-related pain. Overall, analgesic use was low. This study demonstrates the benefits of adopting a systematic and logical approach to the assessment of pain in IPD and the merits of a simple new classification system. PMID- 17085273 TI - An urban community's preferences for hypothetical outcomes of analgesic pain treatment. AB - We assessed preferences of urban residents regarding hypothetical treatment outcomes related to analgesic use to determine how well subjects understood the severity of the outcomes, describe community preferences for these outcomes, and identify predictors of preferences. In a cross-sectional telephone survey, we obtained mean ratings for hypothetical outcomes that included two dimensions of clinical pain (pain severity and potential side effects): A=moderate pain, three side effects; B=mild pain, three side effects; C=moderate pain, one side effect. We focused on 111 respondents who rated Outcome A, moderate pain with three side effects, as the worst condition (the logical choice). Being Spanish speaking predicted preferences across treatment Outcomes A and B. Spanish-speaking subjects and those in fair to poor health tended to view all three outcomes more negatively than other respondents. Knowledge of public preferences can help clinicians better understand factors that influence treatment choices and may help them motivate their patients to adhere to analgesic regimens. PMID- 17085274 TI - Hospice care in the nursing home: changes in visit volume from enrollment to discharge among longer-stay residents. AB - Our objective was to describe the pattern of visits made to longer-stay hospice patients in nursing homes with different diagnoses over the course of their hospice stay. This was a retrospective cohort study in which we used administrative data from a large hospice provider. Longer-stay was defined as hospice enrollment for 12-24 weeks. Residents were categorized by primary diagnosis (cancer, dementia, debility, and other). Three periods of care were isolated: the first week, the penultimate month to death/discharge, and the final week. Visit volume was measured by the number of visits per 100 resident-days. Visits by four provider types were examined: physicians, nurses, aides, and ancillary staff (social workers and chaplains). Visit volume rates were calculated for each time period and were stratified by diagnostic group and provider type. We found that nurses made the most visits and physicians the least. Except for aides, all providers had more activity in the first and final days. Aides' activity increased following the first 2 days and was stable through the remainder of the hospice stay. During the penultimate month, aides had the highest visit rates. Visit rates were similar for all diagnostic groups until the final week of care, when residents with cancer received more visits than others. These patterns of visit volume and configuration confirm that care is most expensive in the earliest and the latest days. PMID- 17085275 TI - You won't know if you're improving unless you measure: recommendations for evaluating Hospice-Veteran Partnerships. AB - Growing evidence indicates that there are abundant opportunities to improve the care that patients receive near the end of life. Hospice care has been associated with improvements in these and other outcomes, but hospice is underused by most patient populations. Therefore, the Department of Veterans Affairs (VA) has made hospice access a priority in its plan to improve end-of-life care for all veterans. In addition to committing funding for hospice care, the VA has also established a national network of Hospice-Veteran Partnerships (HVPs) whose goal is to improve access to hospice for veterans. This article describes the results of a nationwide consensus project to develop measures of the success of HVPs and recommends key measures that should be used to track improvements and to identify opportunities for highly successful collaborative strategies. PMID- 17085276 TI - A case series of patients using medicinal marihuana for management of chronic pain under the Canadian Marihuana Medical Access Regulations. AB - The Canadian Marihuana Medical Access Regulations (MMAR) program allows Health Canada to grant access to marihuana for medical use to those who are suffering from grave and debilitating illnesses. This is a report on a case series of 30 patients followed at a tertiary care pain management center in Nova Scotia who have used medicinal marihuana for 1-5 years under the MMAR program. Patients completed a follow-up questionnaire containing demographic and dosing information, a series of 11-point numerical symptom relief rating scales, a side effect checklist, and a subjective measure of improvement in function. Doses of marihuana ranged from less than 1 to 5g per day via the smoked or oral route of administration. Ninety-three percent of patients reported moderate or greater pain relief. Side effects were reported by 76% of patients, the most common of which were increased appetite and a sense of well-being, weight gain, and slowed thoughts. Limitations of the study include self-selection bias, small size, and lack of a control group. The need for further study using controlled trials is discussed along with an overview of the MMAR program. PMID- 17085277 TI - Metastatic acetabular fractures: evaluation and approach to management. AB - Although bone metastasis to the acetabulum can cause significant disability from pain and immobility, little has been written about the diagnosis and management of a pathologic acetabular fracture. We present three patients with metastatic acetabular fractures and discuss an approach to evaluation and management. When a high index of suspicion of fracture exists, further radiographic workup is warranted. Management requires a multidisciplinary approach. Factors such as age, associated comorbidities, natural history of the underlying primary cancer, general health status, prognosis, acetabular fracture characteristics, and quality of bone should be considered. We briefly discuss the options available to nonoperative candidates. PMID- 17085280 TI - Differential diagnosis of angioedema. AB - There are many conditions that may present with swelling that mimics angioedema. When swelling persists for greater than a few days or is unresponsive to treatment for urticaria/angioedema, other etiologies should be considered. In most instances, a thorough history and physical examination will define other etiologies. However, for more persistent conditions, further laboratory evaluation and a biopsy may be required to define the diagnosis. Rarely is a more aggressive approach required to make the diagnosis. Clinicians should remember that if the swelling does not act like angioedema, it more than likely is not angioedema. PMID- 17085281 TI - Structure and function of C1-inhibitor. AB - C1-INH belongs to the family of serpins. Structural studies have yielded a clear understanding of the biochemical principle underlying the functional activities of these proteins. Although the crystal structure of C1-INH has yet to be revealed, homology modeling has provided a three-dimensional model of the serpin part of C1-INH. This model has helped us understand the biochemical consequences of mutations of the C1-INH gene as they occur in patients who have HAE. The structure of the N-terminal domain of C1-INH remains unknown; however, this part of the molecule is unlikely to be important in the inhibitory activity of C1-INH toward its target proteases. Mutations in this part have not been described in patients who have HAE, except for a deletion containing two cysteine residues involved in the stabilization of the serpin domain. Recent studies suggest some anti-inflammatory functions for this N-terminal part, possibly explaining the effects of C1-INH in diseases other than HAE. PMID- 17085282 TI - Mechanism of angioedema in first complement component inhibitor deficiency. AB - Since shortly after the discovery that hereditary angioedema resulted from deficiency of first complement component (C1) inhibitor, the characterization of the mediator of angioedema has been a major goal. However, because C1 inhibitor regulates activation of both the contract and complement systems, identification of the mediator was not immediately accomplished. For a number of years, some studies appeared to indicate involvement of one system, whereas other studies suggested involvement of the other. However, the vast majority of the evidence accumulated over the past years indicates quite clearly that the major mediator is bradykinin. Therefore, unregulated contact system activation is the defect that leads directly to the development of angioedema. PMID- 17085283 TI - Hereditary angioedema: the clinical syndrome and its management in the United States. AB - There have been important breakthroughs in the understanding and treatment of hereditary angioedema (HAE). An associated abnormality of the serum protein C1 inhibitor led to purified protein use to end attacks. Consideration of the endocrine functions led to rediscovery of impeded androgen use in disease prophylaxis. Considerations of pathophysiology led to introduction of epsilon aminocaproic and tranexemic acids in prophylaxis and to a resurgence in trials of new therapeutic agents. We have gone from a situation where it was not uncommon for patients to have a severe attack sometime in their lives that led to airway compromise and possible death to a situation where death from disease is highly unusual. Thus HAE is in many ways a success story of modern medicine. PMID- 17085284 TI - Acquired deficiency of the inhibitor of the first complement component: presentation, diagnosis, course, and conventional management. AB - Acquired deficiency of the inhibitor of the first complement component (C1-INH) is a rare, potentially life-threatening disease whose cause, course, and management are not completely defined. This article analyzes the etiopathogenetic mechanism, the clinical presentation, and the relationship between acquired C1 INH deficiency and lymphoproliferative disorders. Moreover, the authors give an overview of the outcome of the disease and the different therapies proposed to cure it. PMID- 17085285 TI - Novel therapies for hereditary angioedema. AB - Advances in our understanding of the molecular mechanisms underlying hereditary angioedema (HAE) have led to the development of new treatment modalities. Five new drugs for the treatment of HAE are currently undergoing clinical testing in the United States. These novel therapeutics can be divided into two groups: drugs that replace C1 inhibitor (C1INH) functional activity and drugs that abrogate the bradykinin-mediated increase in vascular permeability associated with HAE attacks. The first group includes two plasma-derived C1INH concentrates as well as a recombinant transgenic human C1INH protein, and the second group includes an engineered plasma kallikrein inhibitor as well as a B2 bradykinin receptor antagonist. This article reviews the rationale, development, and potential use of these novel therapeutics. PMID- 17085286 TI - Hereditary angioedema with normal C1 inhibitor activity including hereditary angioedema with coagulation factor XII gene mutations. AB - Until recently it was assumed that hereditary angioedema is a disease that results exclusively from a deficiency of the C1 inhibitor. In 2000, families with hereditary angioedema, normal C1 inhibitor activity, and protein in plasma were described; all patients were women. In many of the affected women, oral contraceptives, hormone replacement therapy containing estrogens, and pregnancies triggered the clinical symptoms. Recently, in some families mutations in the coagulation factor XII (Hageman factor) gene were detected in the affected women. PMID- 17085287 TI - Angiotensin-converting enzyme inhibitor-associated angioedema. AB - Angioedema, characterized by swelling of the lips, face, and tongue, occurs in anywhere from 0.1% to 6% of angiotensin-converting enzyme (ACE) inhibitor users. The incidence is more common in black Americans than in white Americans, in women than in men, and in smokers than in nonsmokers. The remitting and relapsing nature of ACE inhibitor-associated angioedema can confound clinical recognition of the adverse event but also provides clues to its causes. Defective degradation of vasoactive peptide substrates of ACE, such as bradykinin or substance P, may contribute via non-ACE pathways to the pathogenesis of ACE inhibitor-associated angioedema. PMID- 17085288 TI - Idiopathic recurrent angioedema. AB - The treatment of idiopathic, chronic, recurrent angioedema with or without urticaria is difficult, both for patients and their physicians, because treatment often is only partially effective and is labor-intensive, expensive, and lengthy. New medications for urticaria and angioedema that currently are being tested clinically may prove effective. PMID- 17085289 TI - Anaphylactic and anaphylactoid causes of angioedema. AB - Anaphylactic and anaphylactoid (pseudoallergic) reactions can be expected to occur with greater frequency as the number of immunomodulators are employed. The immune system will become sensitized to these new therapeutic agents or there may be first-dose reactions depending on the pathogenetic mechanism involved. Physicians should review their office or procedure room emergency preparedness protocols and medications. The lack of penicillin major and minor determinants for penicillin testing has made management of penicillin and cephalosporin allergic patients more complicated. In the absence of skin-testing materials, test-challenges will be necessary and performed with less comfort because of not knowing the current level of immunologic sensitization to penicillin. The indication for readministration of any incriminated medication/therapeutic agent should be reviewed. Often, there are not suitable alternatives. Various approaches have been presented to permit safer readministration of essential medications or diagnostic agents to prevent episodes of anaphylaxis or upper airway angioedema. PMID- 17085290 TI - Cytokine-associated angioedema syndromes including episodic angioedema with eosinophilia (Gleich's Syndrome). AB - Angioedema can be associated with many disorders and the presentation can be variable. Subsets of the angioedema syndromes are thought to be cytokine mediated (Table 1). Of these, the best described are the episodic angioedema with eosinophilia syndrome (Gleich's syndrome) and non-episodic angioedema with eosinophilia, which share some common features, but appear to have differences in pathophysiology. NERDS (nodules, eosinophilia, rheumatism, dermatitis and swelling), Clarkson syndrome (idiopathic capillary leak syndrome), and angioedema associated with aldesleukin (human recombinant IL-2) and IFN-alpha have also been reported in the literature, and have been discussed in this review. There is still much to be learned about the pathophysiology, diagnosis, and treatment of patients with these disorders. Our hope is that this review will be of help to those readers who care for patients with these disorders, and will stimulate interest in further research into the pathophysiology of these conditions. PMID- 17085291 TI - Antidiabetic effect of garlic (Allium sativum L.) in normal and streptozotocin induced diabetic rats. AB - OBJECTIVE: The antidiabetic effect of garlic ethanolic extract (Allium sativum L.) was investigated in normal and streptozotocin-induced diabetic rats. RESEARCH METHODS AND PROCEDURE: In the present study, oral administration of garlic extract (0.1, 0.25 and 0.5 g/kg body wt.) for 14 days on the level of serum glucose, total cholesterol, triglycerides, urea, uric acid, creatinine, aspartate amino transferase (AST) and alanine amino transferase (ALT) in normal and streptozotocin-induced diabetic rats were evaluated. RESULTS: Oral administrations of the garlic extract significantly decreased serum glucose, total cholesterol, triglycerides, urea, uric acid, creatinine, AST and ALT levels, while increased serum insulin in diabetic rats but not in normal rats (p<0.05). A comparison was made between the action of garlic extract and glibenclamide (600 microg/kg), the known antidiabetic drug. The antidiabetic effect of the extract was more effective than that observed with glibenclamide. CONCLUSION: It is concluded that the plant must be considered as excellent candidate for future studies on diabetes mellitus. PMID- 17085292 TI - Immunomodulatory effects of arabinogalactan-proteins from Baptisia and Echinacea. AB - The influences of different arabinogalactan-proteins (AGPs) on proliferation and IgM-production of mouse lymphocytes as well as nitrite- and IL6-production of mouse macrophages were investigated in vitro. AGPs have been isolated and purified from roots of Baptisia tinctoria and Echinacea pallida and suspension culture of Echinacea purpurea. Comparing the AGPs, there are differences with regard to fine structure as well as to activities. AGPs from roots of B. tinctoria and E. pallida show high activity in all test systems. AGP from cell culture of E. purpurea shows no influence on proliferation of mouse lymphocytes, only weak influence on the IgM-production of mouse lymphocytes and weak stimulation of nitrite- and IL6-production in alveolar mouse macrophage culture. PMID- 17085293 TI - The effect of erythrodiol-3-acetate on the expressions of matrix metalloproteinase-1 and type-1 procollagen caused by ultraviolet irradiated cultured primary old aged human skin fibroblasts. AB - Methanol and aqueous extracts of Styrax japonica used traditionally for the treatments of skin elastic materials were screened in vitro for the matrix metalloproteinase (MMP)-1 inhibitor actions. The methylene chloride soluble fraction of methanol extract from the stems of S. japonica showed significant MMP 1 inhibition in primary old aged human skin fibroblasts caused by ultraviolet (UV) irradiation. Main triterpenoids were isolated by repeated column chromatography. Among them, the triterpenoid erythrodiol-3-acetate reduced the expression of MMP-1 and induced the expression of type-1 procollagen at the protein levels in a dose-dependent manner caused by UV irradiated cultured old aged human skin fibroblasts. Taken together, our results suggest that erythrodiol 3-acetate plays an important role in the skin aging process caused by UV irradiation. PMID- 17085294 TI - Pharmacological studies of the large-scaled purified genistein from Huaijiao (Sophora japonica-Leguminosae) on anti-osteoporosis. AB - In this report, we used genistein that was extracted from a Chinese herbal medicine Huaijiao (Sophora japonica-Leguminosae) to evaluate its pharmacological function on anti-osteoporosis. This genistein is purified in a large-scale production from Huaijiao by a state-of-art method as described by Tian et al. [2004. The preparation of genistein and LC-MS/MS on-line analysis. Drug Devel. Res. 61, 6-12]. Chemical structure of the isolated genistein was examined by using various techniques including nuclear magnetic resonant spectrum, infrared absorption spectrum, ultraviolet absorption spectrum and mass spectrum, and was proved to be identical to those purified from soybean in a small scale as previously reported. We randomly divided female SD rats into 6 groups, including control, ovariectomized model, Nilestriol-treated, and three level of dosages of genistein-treated. We evaluated the pharmacological effects of genistein against osteoporosis by measuring the bone density of femur and bone mineral group including calcium, phosphorous, and magnesium. The consequences of genistein treatment on bone histology and morphology were also determined by measuring the trabcular area, thickness and number. Our results indicated that treatment with a 4.5 or 9 mg/kg dosage of genistein could also prevent osteoporosis significantly at the 4th week after treatment. In comparison with the anti-osteoporosis effects of soybean genistein, the genistein extracted from Huaijiao has the same beneficial effect on anti-osteoporosis. PMID- 17085295 TI - Antimicrobial activity of aqueous extracts of Larrea divaricata Cav (jarilla) against Helicobacter pylori. AB - We studied here the effect of aqueous extracts of Larrea divaricata Cav on the growth of Helicobacter pylori. Results show that cold extract, infusion, decoction and simulated digestion had inhibitory activity at 0.04-0.1 mg/l against clarithromycin and metronidazole susceptible and resistant H. pylori strains. These results support the popular use of L. divaricata Cav in gastric disturbances and prompt further research to characterize these compounds with a therapeutic potential against gastric ulcers and gastric cancer associated with H. pylori. PMID- 17085296 TI - Effects of Leontice smirnowii tuber monodesmosides and crude extract in carrageenan- and histamine-induced acute inflammation model of rats. AB - Leontice smirnowii is a member of the Berberidaceae family. We have recently reported the 1,1-diphenyl-2-picryl-hydrazyl radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, reducing power and metal chelating activities of L. smirnowii products. In the current study we investigated the possible effects of the crude extracts of L. smirnowii (CELS) and the monodesmoside's purified extract (MPE) of L. smirnowii in the carrageenan and histamine-induced acute inflammation models in rats. The experiment revealed that CELS and MPE have anti-inflammatory effects, dose dependently in carrageenan induced acute inflammation. On the other hand, their proinflammatory effects were surprisingly observed, especially in low doses, in the histamine-induced acute inflammation model. Summarizing these data, we may state that CELS and MPE exert their anti-inflammatory effects via non-histaminergic pathways. PMID- 17085297 TI - In vitro antiprotozoal activity of the lipophilic extracts of different parts of Turkish Pistacia vera L. AB - Thirteen lipophilic extracts prepared with n-hexane from various parts of Pistacia vera L. tree (Anacardiaceae) growing in Turkey were screened for their in vitro activity against four parasitic protozoa, Trypanosoma brucei rhodesiense, Trypanosoma cruzi, Leishmania donovani and Plasmodium falciparum. Melarsoprol, benznidazole, miltefosine, artemisinin and chloroquine were used as reference drugs. The cytotoxic potentials of the extracts on rat skeletal myoblast (L6) cells were also assessed and compared to that of podophyllotoxin. The screening method employed was medium-throughput, where the extracts were tested at two concentrations, at 0.8 and 4.8 microg/ml (T. brucei rhodesiense, L. donovani and Plasmodium falciparum), or at 1.6 and 9.7 microg/ml (T. cruzi and L6 cells). At 4.8 microg/ml concentration, the branch extract of Pistacia vera (PV BR) significantly inhibited (77.3%) the growth of L. donovani, whereas the dry leaf extract (PV-DL) was active against Plasmodium falciparum (60.6% inhibition). The IC50 values of these extracts were determined as 2.3 microg/ml (PV-BR, L. donovani) and 3.65 microg/ml (PV-DL, Plasmodium falciparum). None of the extracts possessed cytotoxicity on mammalian cells. PMID- 17085298 TI - Interpretation of visual field defects respecting the vertical meridian and not related to distinct chiasmal or postchiasmal lesions. AB - Hemianopia respecting the vertical meridian generally results from a disturbance in the optic chiasm or the postchiasmal visual pathway. We present five unusual patients with visual field defects respecting the vertical meridian that were not related to distinct chiasmal or postchiasmal lesions, as determined by magnetic resonance imaging (MRI). Optic neuritis, plus the influence of the testing algorithm for perimetry, was a possible cause in two cases. The symptoms of one patient with homonymous hemianopia were a functional deficit. In the two other cases, the visual field defects may have been caused by optic disc abnormalities. Although visual field defects respecting the vertical meridian, and without any evidence of distinct disease, are uncommon, neurologists and neuro ophthalmologists should consider the differential diagnosis if MRI is negative for distinct lesions in the optic chiasm or the postchiasmal visual pathway. PMID- 17085299 TI - Enhanced hippocampal neurodegeneration after traumatic or kainate excitotoxicity in GFAP-null mice. AB - Astrocytes perform a variety of functions in the adult central nervous system. Recent evidence suggests that the upregulation of glial fibrillary acidic protein (GFAP), an astrocyte-specific intermediate filament component, is a biological marker of neurotoxicity after cerebral injury. We herein compared the response to traumatic brain injury or kainic acid (KA)-induced neurotoxicity in GFAP knockout (GFAP-KO) and wild-type (WT) mice. Seventy-two hours after injury, all GFAP-KO mice showed hippocampal CA3 neurodegeneration, whereas WT mice did not show neurodegeneration. Seventy-two hours after KA administration, GFAP-KO mice were more susceptible to KA-induced seizures and had an increased number of pyknotic damaged CA3 neurons than did WT mice. These results indicate that GFAP plays a crucial role in pyramidal neuronal survival after injury or KA-induced neurotoxicity. PMID- 17085300 TI - Oro-bucco-lingual dyskinesia associated with nonketotic hyperglycaemia. AB - Nonketotic hyperglycaemia is typically associated with a hemiballism-hemichorea pattern of involuntary movement and is characterized by hyperintense signal within the contralateral striatum on T(1)-weighted magnetic resonance imaging (MRI). An oro-bucco-lingual syndrome and location of the lesions has not been previously described. Herein, we report a patient with oro-bucco-lingual dyskinesia associated with nonketotic hyperglycaemia. MRI revealed characteristic T(1) hyperintense lesions in the bilateral putamen. PMID- 17085301 TI - Metastatic adenocarcinoma mimicking 'target sign' of cerebral tuberculosis. AB - We present a 74-year-old male ex-smoker presenting with a 6-week history of personality change, confusion and headache. Magnetic resonance imaging revealed multiple supratentorial and infratentorial parenchymal masses, predominately in the frontal and parietal lobe white matter. A thin enhancing halo was demonstrated with central low signal intensity on T(1)-and T(2)-weighted imaging compatible with calcification. A tiny extra-axial lesion was also noted near the right cerebellopontine angle. Computed tomography (CT) scan confirmed the finding of a 'target' lesion with a central core of calcification and a ring of enhancement. The 'target sign' of intracerebral tuberculomata was first described in 1979 and reported to be pathognomic for this diagnosis in 1988. However, cerebral tuberculosis was considered unlikely clinically because the patient had recently completed a 12-month course of therapy for Mycobacterium avium complex respiratory infection with agents also active against Mycobacterium tuberculosis. He was afebrile and blood tests did not support an inflammatory process. Subsequent histopathology demonstrated metastatic papillary adenocarcinoma and immunohistochemical studies revealed the origin to be that of primary lung carcinoma. A spiculated pulmonary nodule was seen on CT scan but previous bronchoscopy failed to demonstrate malignant cells. In summary, the 'target sign' is a non-specific radiologic finding but most commonly indicates cerebral tuberculoma or metastatic adenocarcinoma in the appropriate clinical context. PMID- 17085302 TI - Clinical applications of high-density surface EMG: a systematic review. AB - High density-surface EMG (HD-sEMG) is a non-invasive technique to measure electrical muscle activity with multiple (more than two) closely spaced electrodes overlying a restricted area of the skin. Besides temporal activity HD sEMG also allows spatial EMG activity to be recorded, thus expanding the possibilities to detect new muscle characteristics. Especially muscle fiber conduction velocity (MFCV) measurements and the evaluation of single motor unit (MU) characteristics come into view. This systematic review of the literature evaluates the clinical applications of HD-sEMG. Although beyond the scope of the present review, the search yielded a large number of "non-clinical" papers demonstrating that a considerable amount of work has been done and that significant technical progress has been made concerning the feasibility and optimization of HD-sEMG techniques. Twenty-nine clinical studies and four reviews of clinical applications of HD-sEMG were considered. The clinical studies concerned muscle fatigue, motor neuron diseases (MND), neuropathies, myopathies (mainly in patients with channelopathies), spontaneous muscle activity and MU firing rates. In principle, HD-sEMG allows pathological changes at the MU level to be detected, especially changes in neurogenic disorders and channelopathies. We additionally discuss several bioengineering aspects and future clinical applications of the technique and provide recommendations for further development and implementation of HD-sEMG as a clinical diagnostic tool. PMID- 17085303 TI - Cutaneous manifestations of chronic graft-versus-host disease. AB - Cutaneous chronic graft versus host disease has traditionally been classified into lichenoid and scleroderma-like forms. However, the initial presentation is sometimes subtle and a variety of less common cutaneous manifestation may be prevalent. This clinical review focuses on the lesional morphology of chronic graft versus host disease, and presents a classification system that may prove useful in early diagnosis. In addition, this approach may help to facilitate the correlation of different morphologic entities with outcome and response to therapy. PMID- 17085304 TI - Minor histocompatibility antigen DDX3Y induces HLA-DQ5-restricted T cell responses with limited TCR-Vbeta usage both in vivo and in vitro. AB - In vitro stimulation of human female T cells with male HLA-identical dendritic cells resulted in the generation of HLA-DQB1*0501/0502-restricted minor histocompatibility H-Y antigen-specific CD4(+) T cell clones. Two clones generated from different HLA-identical pairs were analyzed. Use of HLA-DQ5 expressing female Epstein-Barr virus transformed B lymphoblastoid cell lines transfected with various H-Y genes and loaded with overlapping peptides demonstrated that both T cell clones are specific for a peptide encoded by DDX3Y. Previously, an HLA-DQ5-restricted T cell clone specific for the same peptide was isolated from a patient with graft-versus-host disease. Thus, we compared the T cell receptor (TCR) rearrangements of the 2 in vitro generated T cell clones and the ex vivo isolated T cell clone. All 3 clones shared the same TCRBV5-4* gene segment and 2 of 3 clones also used similar TCR-Valpha segments. Our results suggest that T cells recognizing the HLA-DQ5/DDX3Y T cell epitope might be characterized by a relatively limited TCR-beta repertoire. The differences in the junctional TCR-beta region had no effect on the antigen specificity, but altered the capacity of the TCR to distinguish the HLA-DQ5/DDX3Y complex from its allelic counterpart. The results also demonstrate that in vitro stimulation of T cells with allogeneic HLA-identical dendritic cells may facilitate the characterization of in vivo, potentially relevant HLA class II-restricted minor H epitopes. PMID- 17085305 TI - Gammadelta T cells do not require fully functional cytotoxic pathways or the ability to recognize recipient alloantigens to prevent graft rejection. AB - Gammadelta T cells are a unique and minor T-cell subset that differs from conventional alphabeta T cells by virtue of their tissue localization and antigen processing requirements. We have previously shown that ex vivo-activated gammadelta T cells are able to prevent graft rejection without causing clinically significant graft-versus-host disease (GVHD). In the present study, we examined how gammadelta T cells facilitate alloengraftment and to what extent mechanisms used by conventional alphabeta T cells are also used by gammadelta T cells. We observed that, unlike alphabeta T cells, for which CD8(+) T cells are primarily responsible for facilitating engraftment, purified CD8(+)gammadelta(+) T cells administered at the same fractional dose as for the unseparated activated gammadelta T-cell population were insufficient to prevent graft rejection. Furthermore, the ability to prevent graft rejection was not affected by the absence of fully functional fas ligand or perforin cytotoxic pathways, nor was it contingent on the ability of gammadelta T cells to recognize recipient major histocompatibility process alloantigens. Repetitive infusions of a suboptimal dose of gammadelta T cells however were able to rescue mice from graft rejection, suggesting that the persistence of these cells in vivo was critical in facilitating alloengraftment. These studies demonstrate that gammadelta T cells do not use mechanisms used by conventional nontolerant alphabeta T cells to prevent graft rejection. The ability of these cells to promote engraftment without causing GVHD further distinguishes these cells from alphabeta T cells and may be an attribute that can be exploited in the clinical transplantation setting. PMID- 17085306 TI - Encouraging results with inolimomab (anti-IL-2 receptor) as treatment for refractory acute graft-versus-host disease. AB - Inolimomab [corrected] an anti-interleukin-2 receptor (anti-IL-2R) monoclonal antibody, may be useful in the treatment of steroid-refractory acute graft-versus host disease (aGVHD) by inhibiting 1 of its putative immunopathogenic pathways. We retrospectively analyzed 40 consecutive patients who received inolimomab [corrected] as salvage treatment for steroid refractory aGVHD at a single institution between June 1999 and December 2004. Inolimomab [corrected] was given intravenously at a dose of 11 mg/d for 3 consecutive days, followed by 5.5 mg/d for 7 consecutive days and then 5.5 mg every other day for 5 doses. No infusion related side effects were noted. Twenty-three patients (58%) responded, including 15 (38%) complete and 8 (20%) partial responses. Median overall survival was 294 days (58-996 days) for responders versus 14 days for nonresponders (P < .001), with a 1 year probability of 59% vs 0% for overall survival (P < .0001). Patients without gastrointestinal (GI) involvement showed a higher response rate (100% versus 50% for those without versus with GI involvement, P = .03) In addition, patients who showed some response by day 15 had a higher overall survival (73 +/- 12% vs 24 +/- 12%, respectively, P = .02). The results of this study suggest that inolimomab [corrected] may be an effective salvage therapy for patients with steroid-refractory aGVHD, particularly for those without GI disease, and supports further studies with this agent in prospective controlled trials. PMID- 17085307 TI - Microbial contamination of hematopoietic stem cell products: incidence and clinical sequelae. AB - Microbial contamination of hematopoietic stem cell products is a rare but potentially fatal complication of hematopoietic stem cell transplantation. We report the incidence of contaminated products and describe the clinical outcomes for 35 patients at the University of Minnesota who received contaminated products from January 1990 to December 2004. In total, 2935 products were infused for 2863 transplants during this time, 36 of which 36 (1.2%) were contaminated. Coagulase negative Staphylococcus was the predominant species isolated on culture of the hematopoietic stem cell products. Patients received prophylactic antibiotics before infusion of the contaminated product based on the organism identified from culture and antibiotic sensitivities, if known. After transplantation, blood cultures from 2 patients grew the same pathogen as in the infused contaminated product, including 1 patient who had blood cultures positive for Pseudomonas cepacia. All patients who received contaminated products had benign post transplantation courses except for the patient with Pseudomonas bacteremia, who ultimately died from complications. These results suggest that, although rare, microbial contamination of stem cell products does occur and there must be ongoing efforts by physicians and laboratory personnel to minimize the risk for introduction of contaminants. Prophylactic antibiotics are useful for certain contaminants; however, caution must be exercised when gram-negative contaminated products are administered. PMID- 17085308 TI - Phase I clinical trial of costimulated, IL-4 polarized donor CD4+ T cells as augmentation of allogeneic hematopoietic cell transplantation. AB - The primary objective of this clinical trial was to evaluate the safety, feasibility, and biologic effects of administering costimulated, interleukin (IL) 4 polarized donor CD4(+) T cells in the setting of HLA-matched sibling, T cell replete allogeneic hematopoietic cell transplantation (HCT). Forty-seven subjects with hematologic malignancy received granulocyte colony-stimulating factor mobilized allogeneic hematopoietic cell transplants and cyclosporine graft-versus host disease (GVHD) prophylaxis after reduced intensity conditioning. Initial subjects received no additional cells (n = 19); subsequent subjects received additional donor CD4(+) T cells generated ex vivo by CD3/CD28 costimulation in medium containing IL-4 and IL-2 (administered day 1 after HCT at 5, 25, or 125 x 10(6) cells/kg). Studies after HCT included measurement of monocyte IL-1alpha and tumor necrosis factor alpha, detection of T cells with antitumor specificity, and characterization of T cell cytokine phenotype. The culture method generated donor CD4(+) T cells that secreted increased T helper 2 (Th2) cytokines and decreased T helper 1 (Th1) cytokines. Such Th2-like cells were administered without infusional or dose-limiting toxicity. The Th2 cohort had accelerated lymphocyte reconstitution; both cohorts had rapid hematopoietic recovery and alloengraftment. Acute GVHD and overall survival were similar in the Th2 and non Th2 cohorts. Th2 cell recipients tended to have increased monocyte IL-1alpha and had increased tumor necrosis factor alpha secretion. CD8(+) T cells with antitumor specificity were observed in Th2 and non-Th2 cohorts. Post transplantation T cells from Th2 cell recipients secreted IL-4 and IL-10 (Th2 cytokines) and IL-2 and interferon gamma (Th1 cytokines). Allograft augmentation with costimulated, IL-4-polarized donor CD4(+) T cells resulted in activated Th1, Th2, and inflammatory cytokine pathways without an apparent increase in GVHD. PMID- 17085309 TI - Allogeneic hematopoietic cell transplantation following reduced intensity conditioning for treatment of myelofibrosis. AB - This report describes our experience with reduced intensity conditioning (RIC) allogeneic hematopoietic cell transplantation (HCT) using matched sibling and unrelated donors (MUDs) for treatment of myelofibrosis (MF). Nine patients with MF (median age, 54 years) were treated with RIC allogeneic HCT using MUDs for 7 of the 9 patients and sibling donors for 2 patients. By the Lille classification, 4 patients were characterized as having high risk, 4 as having intermediate risk, and 1 as having low risk. The RIC regimen consisted of fludarabine and a single dose of total body irradiation for the first patient and fludarabine/melphalan for the remaining 8 patients. Granulocyte colony-stimulating factor-primed peripheral blood stem cells (PBSCs) were used for all but 1 patient who received a total of 3 products because of graft failure, of which 2 were bone marrow cells and the third was PBSCs. Prophylaxis against graft-versus-host disease consisted of cyclosporin/mycophenolate with or without methotrexate. Seven patients were successfully engrafted with white blood cells, with an absolute neutrophil count > or =500 by a median of day +15 (range, 10-21 days). At the time of final fluorescence in situ hybridization and/or short tandem repeat analysis, 8 of 9 patients were chimeric, with 96%-100% donor cells and/or DNA. Five of the 9 patients were alive at the time of final contact, with a median follow-up of 32.2 months for the living patients. Overall survival probability at 1 year was 55.6% (95% confidence interval, 31.3%-77.4%). These results suggest that RIC MUD HCT using PBSCs can be an effective treatment for older patients with MF. PMID- 17085310 TI - Human leukocyte antigen DR15 is associated with reduced relapse rate and improved survival after human leukocyte antigen-identical sibling hematopoietic stem cell transplantation. AB - Human leukocyte antigen (HLA) DR15 is associated with autoimmune cytopenia in patients with aplastic anemia, myelodysplastic syndrome, and paroxysmal nocturnal hemoglobinuria. Presence of this antigen also predicts response to immunosuppressive treatment. If DR15 expression on hematopoietic cells also favors induction of immune responses in an allogeneic setting, a lower relapse rate after hematopoietic stem cell transplantation (HSCT) might result through an enhanced graft-versus-leukemia effect. We retrospectively analyzed outcome of HLA identical sibling HSCT in 192 consecutive patients with acute or chronic leukemia or non-Hodgkin lymphoma. Patients carrying the DR15 antigen had a higher estimated 5-year overall survival (76%) than did DR15-negative patients (55%; P = .04). Improved survival for DR15 patients was due to a significant decrease in death from relapse (5% for DR15(+) versus 24% for DR15(-); P = .02), whereas no difference was seen for rates of transplant-related mortality (19% and 21%, respectively; P = .76). Findings were confirmed by multivariate analyses. Our results show an association of DR15 with a decreased risk of disease relapse and improved survival after HSCT for leukemia or non-Hodgkin lymphoma. This adds to the growing list of links between DR15 and immune reactions in hematopoiesis. PMID- 17085311 TI - Unrelated donor status and high donor age independently affect immunologic recovery after nonmyeloablative conditioning. AB - The risk of cytomegalovirus (CMV) infection is higher after HLA-matched unrelated donor (URD) than after HLA-matched related donor (MRD) nonmyeloablative hematopoietic cell transplantation (HCT). We therefore investigated factors affecting immune recovery in 94 patients given HCT from MRDs (n = 51) and URDs (n = 43) after 2-Gy total body irradiation with or without fludarabine and postgrafting immunosuppression with mycophenolate mofetil and cyclosporine. CD4 T cells counts remained below normal values during the first year after HCT in both patient groups. This included abnormally low counts each of naive CD4 T cells and memory CD4 T cells. Conversely, CD8 T cell counts reached the 10th percentile of normal 6 months after HCT in MRD and URD recipients. On day 30 after HCT, URD recipients had lower counts of B cells (P = .02), naive CD4 T cells (P = .04), memory CD4 T cells (P = .005), memory CD8 T cells (P = .005), and CMV-specific T helper cells (P = .007) than had MRD recipients. This delay in CMV-specific immune reconstitution translated into increased frequency of CMV antigenemia among URD recipients during the first 100 days after HCT. Older donor age was associated with low counts of naive CD4 T cells on days 180-365 after HCT (P = .003). Further, low numbers of T cells and CD34(+) cells in the graft and development of acute graft-versus-host disease were associated with impaired immune recovery of naive CD4 T cells and B cells. In summary, immunologic recovery was poor the first year after nonmyeloablative conditioning and was delayed among URD recipients in comparison with MRD recipients. Other factors significantly associated with delayed immune recovery were advanced donor age, low numbers of CD34 and T cells in the graft, and development of graft-versus host disease. PMID- 17085312 TI - A low incidence of nontuberculous mycobacterial infections in pediatric hematopoietic stem cell transplantation recipients. AB - Hematopoietic stem cell transplantation (HSCT) is being used to treat a wide spectrum of clinical disorders but opportunistic infection remains an important factor determining outcomes for these patients. Nontuberculous mycobacterial (NTM) infections are being reported more frequently in HSCT recipients and the incidence of NTM infections in adult recipients is reported to be 0.4%-4.9%. However, the incidence and severity of NTM infections are less well described in pediatric HSCT recipients. Centers for Disease Control and Prevention guidelines were used to define definite and probable NTM infection among 132 children undergoing 169 HSCT between January 2000 and December 2004 at our institution. NTM infection was diagnosed in 5 of 132 pediatric recipients (3.8%). There were no NTM infections diagnosed in the autologous HSCT recipients and the incidence of NTM in allogeneic HSCT recipients was 6.4% (95% confidence interval, 0.8 11.9). The mean age of the HSCT recipients who developed NTM infections was 8 years (range, 2-19 years); 3 were male and 2 were female. Four conditioning regimens included alemtuzumab and 3 had antithymocyte globulin. Of the 5 patients with NTM infections, 2 met the criteria for definite infection and 3 for probable infection. Of the 2 patients with definite NTM infection, 1 had disseminated disease with Mycobacterium avium complex and the other had Mycobacterium chelonae catheter-related bloodstream infection. The probable NTM infections were 1 skin infection with Mycobacterium kansasii and 2 lower respiratory tract infections with M avium complex. Median time to NTM infection was 115 days (range, 14-269 days) after HSCT. Two patients had graft-versus-host disease at the time of NTM infection. All 5 patients received 3-4 antimycobacterial drugs and all NTM infections resolved. In summary, the incidence of NTM infection in pediatric HSCT recipients appears similar to that described in adult HSCT recipients and the outcome appears to be excellent with the proper antibiotic therapy. The increased use of anti-T cell antibodies appears to be associated with an increased risk of NTM infections in pediatric HSCT recipients. Multicenter studies are needed to identify the risk factors, early diagnostic criteria, and optimal therapy. PMID- 17085313 TI - Pegylated TNF-alpha receptor does not prevent acute graft-versus-host disease in the dog leukocyte antigen-nonidentical unrelated canine model. PMID- 17085314 TI - Rituximab responsive refractory acute graft-versus-host disease. PMID- 17085315 TI - Post-transplant lymphoproliferative disorder mimicking a thrombotic microangiopathy. PMID- 17085316 TI - Abstracts and summary of the 4th Annual International Umbilical Cord Blood Transplantation Symposium, Los Angeles, California, May 19-20, 2006. PMID- 17085317 TI - Preparing for the unthinkable: emergency preparedness for the hematopoietic cell transplant program. AB - Emergencies come at the most inopportune times. Some are totally unexpected, others come with short warnings. Although rare, an emergency comes with the potential to wreak disaster and pose serious risk to the services provided to hematopoietic cell transplant (HCT) patients. Yet, the consequences of an emergency can be mitigated by thinking the unthinkable and having a plan for emergency preparedness in place. Each HCT center should develop a plan containing steps of mitigation, preparedness, response and recovery. This report provides the framework for a HCT-specific emergency preparedness plan that can be used by individual centers to develop customized guidelines on preparing for, responding to, and recovering from an emergency. PMID- 17085318 TI - Endogenous prostaglandins regulate spontaneous contractile activity of uterine strips isolated from non-pregnant pigs. AB - Myometrial strips isolated from non-pregnant pigs show spontaneous contractile activity. In the present study, the involvement of endogenous prostaglandins in regulation of uterine spontaneous contraction was investigated using mechanical, immunohistochemical and biochemical approaches. Immunohistochemical study and Western blot analysis for immunoreactive cyclooxygenase (COX) indicated that COX 1 but not COX-2 was expressed predominantly in the myometrium of non-pregnant pigs in a muscle layer-dependent manner (longitudinal muscle>circular muscle). Pretreatment of uterine strips with indomethacin and selective COX-1 inhibitors (SC-560 and FR122047) significantly reduced both the amplitude and frequency of spontaneous contraction in the longitudinal muscle, but inhibition by COX inhibitors was negligible in the circular muscle. On the other hand, CAY10404, a COX-2 inhibitor, did not change the spontaneous contraction in either of the muscle layers. Pretreatment with SC-560 reduced myometrial PGF(2alpha) and PGE(2) levels. Contractile FP and EP(3) receptors were expressed in a muscle layer dependent manner (longitudinal muscle>circular muscle), similar to the expression pattern of COX-1. In conclusion, endogenous prostaglandins produced by COX-1 regulate spontaneous contractile activity of non-pregnant porcine uterine longitudinal muscle selectively due to the heterogeneous expression of contractile prostanoid receptors and COX-1. PMID- 17085319 TI - Quantification of endocannabinoids in rat biological samples by GC/MS: technical and theoretical considerations. AB - In the last several years, interest has increased significantly about the endocannabinoids anandamide and 2-arachidonylglycerol, two lipid messengers that activate cannabinoid receptors. Quantification of these compounds in biological samples presents numerous technical challenges. Because of their low abundance, endocannabinoids are usually quantified by isotope dilution assays using mass spectrometry coupled to either gas chromatography or high-performance liquid chromatography. Although endocannabinoid levels in biological fluids, such as plasma and cerebrospinal fluid, can be directly determined by these techniques, the complex lipid profile of brain tissue samples mandates purification of lipid extracts before GC/MS analysis; this step is not necessary when using HPLC/MS. We have found that when silica gel chromatography is used for endocannabinoid purification, poor recovery and loss of deuterium from the internal standards lead to inaccurate estimation of endocannabinoid levels. By contrast, purification strategies using C(18) solid-phase extraction permits precise and reproducible GC/MS quantification of endocannabinoids in tissue samples. PMID- 17085320 TI - Role of phosphatidylinositol 4,5-bisphosphate in the activation of cytosolic phospholipase A2-alpha. AB - Cytosolic phospholipase A(2)-alpha (cPLA(2)) plays an important role in the release of arachidonic acid and in cell injury. Activation of cPLA(2) is dependent on a rise in cytosolic Ca(2+) concentration, membrane association via the Ca(2+)-dependent lipid binding (CaLB) domain, and phosphorylation. This study addresses the activation of cPLA(2) via potential association with membrane phosphatidylinositol 4,5-bisphosphate (PIP(2)), including the role of a "pleckstrin homology (PH)-like" region of cPLA(2) (amino acids 263-354). In cells incubated with complement, phorbol myristate acetate+the Ca(2+) ionophore, A23187, or epidermal growth factor+A23187, expression of the PH domain of phospholipase C-delta1 (which sequesters membrane PIP(2)) attenuated cPLA(2) activity. Stimulated cPLA(2) activity was also attenuated by the expression of cPLA(2) 135-366, or cPLA(2) 2-366, and expression of a PIP(2)-specific 5' phosphatase. However, in a yeast-based assay that tests the ability of proteins to bind to membrane lipids, including PIP(2), with high affinity, only cPLA(2) 1 200 (CaLB domain) was able to interact with membrane lipids, whereas cPLA(2)s 135 366, 2-366, 201-648, and 1-648 were unable to do so. Therefore, cPLA(2) activity can be modulated by sequestration or depletion of cellular PIP(2), although the interaction of cPLA(2) with membrane PIP(2) appears to be indirect, or of weak affinity. PMID- 17085321 TI - Concomitant activation of extracellular signal-regulated kinase and induction of COX-2 stimulates maximum prostaglandin E2 synthesis in human airway epithelial cells. AB - The intracellular regulation and kinetics of prostaglandin (PG)E(2) synthesis in human airway epithelial (NCI-H292) cells was investigated. Interleukin (IL) 1beta, tumor necrosis factor (TNF)-alpha and lipopolysaccharide (LPS) all induced PGE(2) synthesis (p<0.001) and transient (5-15 min) phosphorylation of extracellular signal-regulated kinase (ERK). Phorbol myristate acetate (PMA) and calcium ionophore, A23187 further enhanced PGE(2) synthesis (p<0.001) and caused phosphorylation of ERK that was sustained for up to 16 h. COX-2 protein expression and PGE(2) synthesis were increased following exposure to combinations of stimuli that increased intracellular Ca(2+), and activated protein kinase C as well as ERK. Inhibition of ERK almost completely abrogated PGE(2) synthesis in response to all stimuli. Sustained, maximum PGE(2) synthesis was observed when cells were stimulated such that ERK phosphorylation was concomitant with increased COX-2 protein expression. These results argue against redundancy in pathways for PGE(2) synthesis, and suggest that at various stages of inflammation different stimuli may influence ERK activation and COX-2 expression, so as to tightly regulate the kinetics and amount of PGE(2) produced by airway epithelial cells in response to lung inflammation. PMID- 17085322 TI - In vivo evidence that N-oleoylglycine acts independently of its conversion to oleamide. AB - Oleamide (cis-9-octadecenamide) is a member of an emerging class of lipid signaling molecules, the primary fatty acid amides. A growing body of evidence indicates that oleamide mediates fundamental neurochemical processes including sleep, thermoregulation, and nociception. Nevertheless, the mechanism for oleamide biosynthesis remains unknown. The leading hypothesis holds that oleamide is synthesized from oleoylglycine via the actions of the peptide amidating enzyme, peptidylglycine alpha-amidating monooxygenase (PAM). The present study investigated this hypothesis using pharmacologic treatments, physiologic assessments, and measurements of serum oleamide levels using a newly developed enzyme-linked immunosorbant assay (ELISA). Oleamide and oleoylglycine both induced profound hypothermia and decreased locomotion, over equivalent dose ranges and time courses, whereas, closely related compounds, stearamide and oleic acid, were essentially without effect. While the biologic actions of oleamide and oleoylglycine were equivalent, the two compounds differed dramatically with respect to their effects on serum levels of oleamide. Oleamide administration (80mg/kg) elevated blood-borne oleamide by eight-fold, whereas, the same dose of oleoylglycine had no effect on circulating oleamide levels. In addition, pretreatment with the established PAM inhibitor, disulfiram, produced modest reductions in the hypothermic responses to both oleoylglycine and oleamide, suggesting that the effects of disulfiram were not mediated through inhibition of PAM and a resulting decrease in the formation of oleamide from oleoylglycine. Collectively, these findings raise the possibilities that: (1) oleoylglycine possesses biologic activity that is independent of its conversion to oleamide and (2) the increased availability of oleoylglycine as a potential substrate does not drive the biosynthesis of oleamide. PMID- 17085323 TI - Effects of selective PGE2 receptor antagonists in esophageal adenocarcinoma cells derived from Barrett's esophagus. AB - Accumulating evidence suggests that COX-2-derived prostaglandin E(2) (PGE(2)) plays an important role in esophageal adenocarcinogenesis. Recently, PGE(2) receptors (EP) have been shown to be involved in colon cancer development. Since it is not known which receptors regulate PGE(2) signals in esophageal adenocarcinoma, we investigated the role of EP receptors using a human Barrett's derived esophageal adenocarcinoma cell line (OE33). OE33 cells expressed COX-1, COX-2, EP(1), EP(2) and EP(4) but not EP(3) receptors as determined by real time RT-PCR and Western-blot. Treatment with 5-aza-dC restored expression, suggesting that hypermethylation is involved in EP(3) downregulation. Endogenous PGE(2) production was mainly due to COX-2, since this was significantly suppressed with COX-2 inhibitors (NS-398 and SC-58125), but not COX-1 inhibitors (SC-560). Cell proliferation ((3)H-thymidine uptake) was significantly inhibited by NS-398 and SC-58125, the EP(1) antagonist SC-51322, AH6809 (EP(1)/EP(2) antagonist), and the EP(4) antagonist AH23848B, but was not affected by exogenous PGE(2). However, treatment with the selective EP(2) agonist Butaprost or 16,16-dimethylPGE(2) significantly inhibited butyrate-induced apoptosis and stimulated OE33 cell migration. The effect of exogenous PGE(2) on migration was attenuated when cells were first treated with EP(1) and EP(4) antagonists. These findings suggest a potential role for EP selective antagonists in the treatment of esophageal adenocarcinoma. PMID- 17085324 TI - LC-MS/MS-analysis of sphingosine-1-phosphate and related compounds in plasma samples. AB - Sphingosine-1-phosphate (S1P) and related compounds are important signaling molecules and are normal constituents of human plasma. So far, only a few methods exist for their determination specifically in plasma demanding radioactive agents, more or less time consuming extraction or derivatization procedures. Here, we describe a very simple, reliable, sensitive standard-addition method for the simultaneous determination of S1P, sphingosine (SPH), sphinganine (SAPH) and sphinganine-1-phosphate (SA1P) in human and rat plasma samples. After methanol precipitation of plasma samples the supernatants were directly assessed by liquid chromatography-electrospray ionisation-tandem mass spectrometry (LC-ESI-MS/MS). HPLC analysis was done under gradient conditions using a C18 reversed phase column. The lower limit of quantification (LLOQ) was <10.2, <4.6, <1.9 and 0.57ng/ml for S1P, SPH, SAPH and SA1P, respectively. Variations in accuracy and intraday and interday precision were <15% over the range of calibration. All analytes were normal constituents both in human and rat plasma although the SA1P concentrations in a few rat plasma samples were below the lower limit of quantification. This validated method is suitable to generate new pharmacological findings by monitoring plasma concentrations of S1P and related compounds especially when low amounts of plasma samples are present (e.g. plasma samples from rodents). PMID- 17085325 TI - Platelet-activating factor (PAF) is the effector of IFN gamma-stimulated invasiveness and motility in a B16 melanoma line. AB - In this study, we investigated whether PAF synthesized by F10-M3 cells (a clone of B16-F10 melanoma line) mediates the increased capacity of these cells to penetrate into Matrigel upon stimulation with IFN gamma. The determination of PAF synthesized by IFN gamma-stimulated tumor cells revealed that 70% of newly synthesized PAF was released into growth media, while the remaining 30% was associated with the cell bodies. An experimental protocol based on the use of WEB 2086, a PAF receptorial antagonist, was designed to explore which of the two fractions of PAF synthesized by IFN gamma-stimulated F10-M3 cells (released into the growth medium or associated with the cell bodies) is essential to their capacity to migrate through Matrigel. We found that the PAF secreted into growth medium is the fraction responsible for the enhanced invasiveness of melanoma cells stimulated with IFN gamma. We also investigated whether motility of melanoma cells is stimulated by IFN gamma, and, if so, whether PAF is involved in this effect. We found that WEB 2086 prevented the remodeling of stress fibers, examined as an index of cell motility, that we observed in F10-M3 cells stimulated with IFN gamma. Furthermore, the observation that PAF receptor is expressed in IFN gamma-stimulated melanoma cells suggests that the invasive phenotype (e.g. migration through a reconstituted basement membrane and motility) promoted by PAF is based on an autocrine mechanism. On the whole, these results might indicate that PAF contributes to the expression of properties typical of an invasive phenotype in tumor cells stimulated with cytokines. PMID- 17085326 TI - Effects of selective prostaglandins E2 receptor agonists on cultured calvarial murine osteoblastic cells. AB - We compared the direct effects of selective EP4 and EP2 receptor agonists (EP4A and EP2A) with prostaglandin E(2) (PGE(2)) on the differentiation of cultured murine calvarial osteoblastic cells. EP4A increased alkaline phosphatase activity and osteocalcin mRNA levels in these cultures similar to PGE(2). This effect was seen with both "direct plating" immediately after isolating the cells, or "indirect plating" in which the cells were grown to confluence and replated. EP2A had a smaller effect, significant only in "indirect plating" experiments. All three agents decreased the DNA and protein content in indirect plating experiments, but not in direct plating experiments. We conclude that the anabolic effect of PGE(2) in calvarial osteoblastic cell cultures is largely mediated by activation of the EP4 receptor, while activation of the EP2 receptor is less effective. PMID- 17085330 TI - Current status and perspective of liver preservation solutions. AB - BACKGROUND: A safe and effective preservation solution is a precondition for liver transplantation, which is accepted as the radical treatment for patients with end-stage liver disease. The increasing use of marginal donors and non-heart beating donors as well as the establishment of a national organ allocation network call for better preservation. New preservation solutions like histidine tryptophan-ketoglutarate (HTK) solution and Celsior solution have been introduced to liver preservation, and protective gene intervention and other modifications have also been investigated. In this article, we review recent advances in liver preservation solutions. DATA SOURCES: An English-language literature search was conducted using MEDLINE (1990-2005) on liver preservation solution, biliary complication, protective gene and other related subjects. RESULTS: Although the high viscosity of the University of Wisconsin (UW) solution proved harmful to the hepatic microcirculation, three solutions showed equivalent preservation effects. When the cold ischemia time was short, there were no significant differences among the three solutions in the incidence of biliary complications. So far, modifications of preservation solutions have achieved great success. Several types of protective genes like A20, Bcl-2, Bcl-X(L) and HO-1 were reported to have definite liver protective effects. The addition of other substrates like TNF alpha antibody, tacrolimus (FK506) and fructose-1,6-bisphosphate (FBP) can also improve the preservation effect. However, addition of insulin to UW solution is harmful to the graft. CONCLUSIONS: In centers with highly-developed transplantation techniques, HTK and Celsior solutions are acceptable in liver preservation. Protective gene modification and addition of substrates like TNF alpha antibody, FK506 and FBP are prominent approaches to improve liver preservation. PMID- 17085331 TI - Interventional treatments for hepatocellular carcinoma. AB - BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most frequent primary malignant tumors in the world. Hepatic resection and liver transplantation are considered optimal for potential treatment of HCC. However, only 20% of HCCs can be surgically treated. And most of surgically-noneligible patients have to receive interventional managements including local ablation and transarterial chemoembolization (TACE). In this paper, we review the interventional treatments of HCC. DATA SOURCES: A literature search of PubMed database was conducted and research articles were reviewed. RESULTS: Percutaneous ethanol injection (PEI) is usually applied to small HCC for a complete necrosis. Radiofrequency ablation, an alternative to PEI, also causes tumor necrosis and needs fewer times of ablation. Other methods such as acetic acid injection, laser, microwave, etc have enriched local ablation for HCC. High intensity focus ultrasound (HIFU) is thought to be promising. TACE, another common modality, can improve the survival rate of patients with HCC. The newly developed embolic agents and adjuvant rAd-p53 gene therapy are well reported. CONCLUSIONS: Surgically-noneligible HCC can be treated with interventional procedures. Each method has its advantages and disadvantages. However, it is still pressing to develop ablative methods as well as new embolic agents for a better prognosis of HCC. PMID- 17085332 TI - Progress in research into the mechanism of Radix salviae miltiorrhizae in treatment of acute pancreatitis. AB - BACKGROUND: Acute pancreatitis (AP) is a common acute abdomen with high mortality, multiple complications and complicated causes. There is no effective therapy for AP. Radix salviae miltiorrhizae (Danshen), a traditional herbal medicine, has a low price and a wide range of clinical applications. It is effective to promote blood flow, eliminate stagnancy, and relieve pain. It is also found to be effective in treating AP. We reviewed the progress in research into the mechanism of Radix salviae miltiorrhizae in treatment of AP. DATA SOURCES: An English-language literature search was conducted using MEDLINE (1988 2005) on Radix salviae miltiorrhizae (Danshen) and acute pancreatitis. RESULTS: The mechanisms of Danshen in the treatment of AP include improvement of microcirculatory disturbances; elimination of oxygen free radicals; modulation of the metabolism of lipid inflammatory mediator; and blocking of calcium inflow and prevention of calcium overload. CONCLUSION: Danshen can effectively reduce the mortality and complications of AP. PMID- 17085333 TI - Utility of adenovirus-mediated Fas ligand and bcl-2 gene transfer to modulate rat liver allograft survival. AB - BACKGROUND: Expression of Fas ligand (FasL) on the graft by gene transduction is expected to introduce apoptosis to lymphocytes to protect rejection, but the FasL expressing graft cells may also induce apoptosis as the graft usually expresses Fas antigens. In this study, a strong antiapoptotic gene, bcl-2, was cotransfected with the FasL gene in rat liver graft to protect against Fas mediated cell death and to prolong recipient survival. METHODS: Orthotopic liver transplantation was done in a strain combination of DA to LEW rats. After donor vascular isolation, adenovirus-mediated FasL and bcl-2 genes were cotransfected in the liver graft. RESULTS: Intragraft expression of FasL mRNA was constitutively expressed after adenovirus-mediated transduction, although expression of FasL increased mildly in control grafts. Bcl-2 mRNA was highly expressed at 2 days after reperfusion. In contrast, lower expression of bcl-2 was observed in the control group. The average survival of the gene transferred allografts increased from (9.8+1.3) days to (18.5+8.7) days compared with the control group. CONCLUSION: Our results indicate that rat liver allografts can be protected against host immune responses by adenovirus-mediated FasL and bcl-2 transfection, and that bcl-2 expression prevents the graft from Fas-mediated apoptosis. PMID- 17085334 TI - Do the lungs contribute to propofol elimination in patients during orthotopic liver transplantation without veno-venous bypass? AB - BACKGROUND: The clearance of propofol is very rapid, and its transformation takes place mainly in the liver. Some reports indicated extrahepatic clearance of the drug and that the lungs are the likely place where the process occurs. This study was undertaken to compare the plasma concentrations of propofol both in the pulmonary and radial arteries after constant infusion during the dissection, anhepatic and reperfusion phases of orthotopic liver transplantation (OLT) without veno-venous bypass, attempting to investigate extrahepatic clearance and to determine whether the human lungs take part in the elimination of propofol. METHODS: Fifteen patients undergoing OLT without veno-venous bypass were enrolled in the study, and propofol was infused via a forearm vein at a rate of 2 mg x kg 1 x h-1. Blood samples were simultaneously collected from pulmonary and radial arteries at the end of the first hepatic portal dissection (T0), at the clamping of the portal vein (T1), 30, and 60 minutes after the beginning of the anhepatic phase (T2, T3), and 30, 60, and 120 minutes after the unclamping of the new liver (T4, T5, T6). Plasma propofol concentrations were measured using a reversed phase, high-performance liquid chromatographic method with fluorescence detection. RESULTS: The concentrations of plasma propofol in the pulmonary and radial arteries at T2 and T3 rose significantly compared with T0 and T1 (P<0.01) respectively. After reperfusion, the drug concentrations at T4, T5 and T6 decreased significantly compared with T2, T3 (P<0.01) respectively. There were no significant differences in plasma propofol concentrations between the pulmonary and radial arteries at any time points. CONCLUSIONS: Propofol is eliminated mainly by the liver, and also by extrahepatic organs. The lungs seem to be not a major site contributing to the extrahepatic metabolism of propofol in humans. PMID- 17085335 TI - Prolonging warm ischemia reduces the cold preservation limits of liver grafts in swine. AB - BACKGROUND: The critical shortage of transplantable organs necessitates utilization of unconventional donors. But the safe time limits of cold preservation of liver grafts subjected to warm ischemia (WI) for up to 30 minutes from non-heart-beating-donors (NHBDs) has not been delineated. In this study, we investigated how the limits of cold ischemia (CI) in University of Wisconsin (UW) solution are changed in liver grafts subjected to WI from 10 to 30 minutes. METHODS: A simple porcine NHBD liver transplantation (LT) model was developed. In donors, livers were subjected to 10, 20 or 30 minutes of WI and subsequent different times of CI in UW solution. Animals were divided into three groups (WI 10 min, WI 20 min, WI 30 min, n=13 in each group) and nine subgroups (from CI 6 h to CI 28 h). One-week survival rates of recipients, hepatic function, liver energy metabolism, grafted liver microcirculation and pathological observations of the liver were compared. RESULTS: In the WI 10 min group, the one-week survival rate of the CI 20 h subgroup was significantly higher than in the other two subgroups (CI 24 h and CI 28 h) (P<0.05). Furthermore, the CI 20 h subgroup had a lower level of alanine aminotransferase (ALT), less pathological damage, a higher concentration of adenosine triphosphate (ATP) and microcirculatory blood flow in the grafted livers at 1 hour after reperfusion than the other two subgroups. The same trends were also found in the other two groups (WI 20 min and WI 30 min) and their subgroups. CONCLUSIONS: The cold preservation limits of the liver grafts shortened from 20 to 12 to 6 hours when WI time was prolonged from 10 to 20 to 30 minutes. Only the liver grafts within these time limits could be safely transplanted. PMID- 17085336 TI - Energy metabolism and survival of liver grafts from non-heart-beating donor rats with warm ischemia injury. AB - BACKGROUND: The shortage of donor livers is a critical limiting factor for the use of liver transplantation in treatment of end-stage liver diseases. Organs from non-heart-beating donors seem to be an effective option to alleviate this problem. Warm ischemia injury, however, directly influences the grafts' activity and functional recovery after operation. We investigated the energy metabolism and post-transplant survival of liver grafts after different warm ischemia times (WITs) in rats and determined the maximum limit for liver grafts with warm ischemia. METHODS: Rats were randomized into 7 groups with WITs of 0 (control), 10, 15, 20, 30, 45 or 60 minutes. The indices of energy metabolism were measured by reversed-phase high performance liquid chromatograpy and all liver graft specimens were subjected to ultrastructural observation. After orthotopic liver transplantation (OLT), the recovery of energy metabolism in liver grafts after 24 and 48 hours and the survival of the rats were assessed. RESULTS: The levels of adenosine triphosphate (ATP) and energy charge (EC) decreased gradually after different WITs in a time-dependent manner, and this was especially significant within 30 minutes. The levels of ATP and EC in liver grafts with 30 minutes of warm ischemia largely recovered 24 hours after OLT, with 45 minutes of warm ischemia partially recovered 48 hours after OLT, and with 60 minutes of warm ischemia, hardly recovered even 48 hours after OLT. The survival time after OLT did not significantly change with up to 30 minutes of WIT, while long-term survival was reduced with 45 and 60 minutes of WIT. CONCLUSIONS: The levels of ATP and EC after OLT may be important criteria for evaluating the quality of a liver graft. The WIT of a liver graft is closely related to the recovery of hepatic energy metabolism and the graft survival. PMID- 17085337 TI - Resective surgery for liver tumor: a multivariate analysis of causes and risk factors linked to postoperative complications. AB - BACKGROUND: In spite of accurate selection of patients eligible for resection, and although advances in surgical techniques and perioperative management have greatly contributed to reducing the rate of perioperative deaths, stress must be placed on reducing the postoperative complication rates reported to be still as high as 50%. This study was designed to analyze the causes and foreseeable risk factors linked to postoperative morbidity on the grounds of data derived from a single-center surgical population. METHODS: From September 1989 to March 2005, 287 consecutive patients, affected either with HCC or liver metastasis, had liver resection at our department. Among the HCC series we recorded 98 patients (73.2%) in Child-Pugh class A, 32 (23.8%) in class B and 4 in class C (3%). In 104 colorectal metastases, 71% were due to colon cancer, 25% rectal, 3% sigmoid, and 1% anorectal. In 49 non-colorectal metastases, 22.4% were derived from breast cancer, 63.2% gastrointestinal tumors (excluding colon) and 14.4% other cancers. We performed 80 wedge resections, 77 bisegmentectomies and/or left lobectomies, 74 segmentectomies, 22 major hepatectomies, 20 left hepatectomies, and 14 trisegmentectomies. RESULTS: The in-hospital mortality rate in this series was 4.5%, and the morbidity rate was 47.7%, because of pleural effusion (30%), hepatic abscess (25%), hepatic insufficiency (19%), ascites (10%), hemoperitoneum (10%), or biliary fistula (6%). The variables associated with the technical aspects of the surgical procedure that were responsible for the complications were: a Pringle maneuver length more than 20 minutes (P=0.001); the type of liver resection procedure, including major hepatectomy (P=0.02), left hepatectomy (P=0.04), trisegmentectomy (P=0.04), bisegmentectomy and/or left lobectomy (P=0.04); and a blood transfusion of more than 600 ml (P=0.04). CONCLUSION: The evaluation of causes and foreseeable risk factors linked to postoperative morbidity during the planning of surgical treatment should play the same role as other factors weighed in the selection of patients eligible for liver resection. PMID- 17085338 TI - Cisapride decreasing orocecal transit time in patients with nonalcoholic steatohepatitis. AB - BACKGROUND: Altered small-intestine motility, lengthening of orocecal transit time (OCTT), and small-intestinal bacterial overgrowth (SIBO) have been detected in patients with nonalcoholic steatohepatitis (NASH). These changes might be related to the progressive course and poor prognosis of the disease. This study was undertaken to investigate the effect of 4-week treatment with cisapride on OCTT. METHODS: Ten NASH patients without diabetes were included. Ten healthy individuals served as controls. OCTT was measured by lactulose breath test (LBT). Anti-endotoxin core antibodies (EndoCAb) IgG were also examined. The effect of cisapride (10 mg TID during 4 weeks) on LBT and serum EndoCAb IgG levels in NASH patients was investigated. RESULTS: The NASH patients had more significantly prolonged OCTT (95 +/- 17 min) than the controls (59 +/- 18 min, P=0.00032). Cisapride administration decreased OCTT (from 95 +/- 17 min to 83 +/- 19 min, P=0.037), basal breathed H2 (from 9.87 +/- 1.60 ppm to 8.61 +/- 1.63 ppm, P=0.046) and EndoCAb IgG titers (from 5.24 +/- 0.68 GMU/ml to 4.20 +/- 0.72 GMU/ml, P=0.013) in NASH patients. CONCLUSIONS: The present data suggest the existence of deranged intestinal motility and endotoxemia in NASH patients. Cisapride administration during 4 weeks possibly restore intestinal motility and ameliorate endotoxemia in NASH patients. PMID- 17085339 TI - Combined serum hepatoma-specific alpha-fetoprotein and circulating alpha fetoprotein-mRNA in diagnosis of hepatocellular carcinoma. AB - BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide, its prognosis is poor, and early detection is of utmost importance. Although alpha-fetoprotein (AFP) is a useful marker for detecting and monitoring HCC development, the false-negative or false-positive rates with AFP alone may be as high as 30%-40% for patients with small HCCs. To enhance the specificity and accuracy of AFP measurements for HCC, we analyzed AFP expression states in livers, detected the hepatoma-specific AFP (HS-AFP) fraction and AFP mRNA from peripheral blood mononuclear cells, and explored their clinical implications for HCC diagnosis. METHODS: AFP expression and hepatocyte distributions in liver specimens were investigated by an immunohistochemical assay. Total RNAs were extracted from circulating blood, synthesized to cDNA through random primers and reverse transcriptase, and fragments of the AFP gene were amplified by a nested-PCR assay. The HS-AFP fraction was separated by lectin affinity chromatography and its level was detected by radioimmunoassay. RESULTS: The incidence of AFP was 73.3% in HCC tissues and its expression in HCCs with moderate or low differentiation was significantly stronger than that of HCCs with high differentiation (P<0.05). The incidence of AFP gene fragments was 100% in HCCs, and 60% in paracancerous tissues (P<0.01). In the HCC and liver cirrhosis groups, the incidence of HS-AFP was 91.7% and 18% (P<0.01), and of AFP-mRNA was 56.7% and 16% (P<0.01), respectively, and neither was found in controls. HS-AFP or AFP-mRNA was not significantly related to size or number of HCC, but to its differentiation, metastasis, and relapse (P<0.05). CONCLUSIONS: Different AFP expression is present in different parts of HCC tissues. HS-AFP and AFP-mRNA fragments improve sensitivity and specificity, and both are useful markers to diagnose HCC or monitor metastasis and relapse. PMID- 17085340 TI - Therapeutic efficacy and bone marrow protection of the mdr1 gene and over-dose chemotherapy with doxorubicin for rabbits with VX2 hepatocarcinoma. AB - BACKGROUND: Malignant tumors are common diseases threatening to the health and life of human being. Clinically, the multidrug resistance of tumor cells and bone marrow depression caused by chemotherapeutic agents are the main obstacles to the treatment of tumors, and both are related to the mdr1 gene. The over expression of the mdr1 gene in tumor cells contributes to the multidrug resistance of malignant tumor cells. With little expression of the mdr1 gene, bone marrow cells particularly susceptible to multidrug resistance-sensitive agents, which cause serious toxicity in bone marrow. This study was undertaken to assess therapeutic efficacy of transplantation of bone marrow mononuclear cells transferred with the mdr1 gene and over-dose chemotherapy with doxorubicin for VX2 hepatocarcinoma of rabbits. METHODS: The mdr1 gene was transferred into the bone marrow mononuclear cells of rabbits, which was co-cultured with retroviral vector-containing supernatant, and the cells were autotransplanted into a rabbit model with VX2 hepatocarcinoma. After chemotherapy with doxorubicin, the protective effects of the mdr1 gene and therapeutic efficacy of over-dose chemotherapy were observed. RESULTS: The mdr1 gene was transferred successfully into the bone marrow mononuclear cells, with a transduction efficiency of 35%. After autotransplantation, the mdr1 gene was expressed functionally in bone marrow with a positive rate of 8%, indicating that the gene played an important role in bone marrow protection. The rabbits with VX2 hepatocarcinoma, which had received the mdr1 gene-transduced cells, survived after chemotherapy with a 3-fold dose of adriamycin, and their white blood cell counts were (4.26 +/- 1.03) x 10(4)/L. Since hepatocarcinoma cells were eradicated, the survival time (97.00 +/- 46.75 d) of the rabbits was extended (P<0.05) and the healing rate of the tumor was increased (P<0.05). CONCLUSIONS: The transferring of the mdr1 gene into bone marrow mononuclear cells could confer chemoprotection to bone marrow, and over dose chemotherapy could be prescribed for the treatment of malignant tumors. PMID- 17085341 TI - Effects of multidrug resistance, antisense RNA on the chemosensitivity of hepatocellular carcinoma cells. AB - BACKGROUND: Multidrug resistance is a major obstacle in cancer chemotherapy. We examined whether the antisense RNA of multidrug resistance gene 1 (mdr1) could reverse multidrug resistance in the human hepatocellular carcinoma (HCC) cell line SMMC7721/ADM. METHODS: The recombinant adenoviruses pAdEasy-GFP-ASmdr1 product was produced by the adenoviral vector AdEasy system, which can express antisense RNA against the mdr1 gene. Following that, the recombinant adenovirus was transfected into the P-glycoprotein-producing multidrug resistance cell line, SMMC7721/ADM human HCC cells resistant to adriamycin (ADM) and daunorubicin (DNR). In order to investigate the reversal of multidrug resistance phenotype, we measured the expression of mdr1 mRNA by RT-PCR and the production of P glycoprotein by flow cytometry. The sensitivities for ADM and DNR SMMC7721/ADM cells were examined by [3-(4, 5-dimethylthi-azol-2-yl)-2,5 diphenyl-terazolium bromide] (MTT) analysis. RESULTS: The low-level expression of mdr1 mRNA and P glycoprotein production were observed in parental sensitive cells SMMC/7721 in addition to the overexpression of mdr1 mRNA and P-glycoprotein in SMMC7721/ADM cells. The transfection of antisense-RNA into SMMC7721/ADM cells resulted in decreases of mdr1 mRNA and P-glycoprotein, but increase of drug sensitivities. The sensitivities of transfected SMMC7721/ADM cells to ADM and DNR in IC50 reduced by 31.25% and 62.96% respectively. CONCLUSIONS: Mdr1 antisense RNA can increase the sensitivities of SMMC7721/ADM cells to anticancer drug by decreasing the expression of the mdr1 gene and inhibiting P-glycoprotein expression. This strategy may be applicable to cancer patients with P-glycoprotein mediated multidrug resistance. PMID- 17085342 TI - Is the hypoxia-inducible factor-1 alpha mRNA expression activated by ethanol induced injury, the mechanism underlying alcoholic liver disease? AB - BACKGROUND: Excessive alcohol consumption can result in multiple organ injury, of which alcoholic liver disease (ALD) is the most common. With economic development and improvement of living standards, the incidence of diseases caused by alcohol abuse has been increasing in China, although its pathogenesis remains obscure. The aim of this study was to investigate the role of hypoxia in chronic ALD. METHODS: Twenty-eight male Sprague-Dawley rats were randomized into a control group (n=12) with a normal history and an experimental group (n=16) fed with 10 ml/kg of 56% (vol/vol) ethanol once per day by gastric lavage for 24 weeks. At 24 weeks, blood samples were collected and then the rats were killed. Liver samples were frozen at -80 degrees C and used for RT-PCR; other liver samples were obtained for immunohistochemical staining. RESULTS: When the period of alcohol consumption increased, the positive rate of expression of hypoxia-inducible factor-1 alpha (HIF-1alpha) mRNA was more significantly elevated in the liver of the alcohol group than in the control group (P < or = 0.05). The HIF-1alpha protein located in the cytoplasm was seldom expressed in the control group, but significantly in the alcohol group (P < or = 0.01). CONCLUSION: HIF-1alpha mRNA expression was activated by ethanol-induced injury in this study, suggesting that hypoxia is involved in the underlying mechanism of ALD. PMID- 17085343 TI - Influence of nitric oxide synthase and cyclooxygenase blockade on expression of cyclooxygenase and hemodynamics in rats with portal hypertension. AB - BACKGROUND: The importance of nitric oxide (NO) in the pathogenesis of portal hypertension (PHT) has been extensively studied, but whether or not prostacyclin (PGI(2)) plays a role in formation and development of hyperdynamic circutatory state in PHT has not been verified. The present study was undertaken to investigate the possible interaction between prostacyclin (PGI(2)) and nitric oxide (NO) in the hyperdynamic circulatory state of rats with chronic portal hypertension (PHT), by measuring the hemodynamic changes and expression of cyclooxygenase (COX) mRNA in vessels and small intestine after administration of N(omega)-nitro-L-arginine (L-NNA) or indomethacin (INDO) either in the short-term (7 days) or long-term (15 days). METHODS: Ninety-seven male Sprague-Dawley rats were divided into three groups: intrahepatic portal hypertension (IHPH) induced by injection of CCl(4), prehepatic portal hypertension (PHPH) induced by partial stenosis of the portal vein, and sham-operated controls (SO). Animals of each group received L-NNA or INDO either for 7 or 15 days, with saline as control. Splanchnic hemodynamics was measured by the radioactive microsphere technique. The concentration of NO in serum was determined as the nitrate; nitrite ratio (NO(2)(-)/NO(3)(-), micromol/L) by a colorometric method, and that of PGI(2) was measured by specific radioimmunoassay for its stable hydrolysis product 6-keto PGF(1alpha) (pg/ml). The reverse transcription-polymerase chain reaction measured the levels of COX-1 mRNA in the superior mesenteric artery, thoracic aorta, and small intestine of these rats. RESULTS: Compared with SO rats, COX-1 mRNA expression and the concentrations of plasma 6-keto-PGF(1alpha) and serum NO(2)( )/NO(3)(-) were enhanced in both IHPH and PHPH rats; splanchnic vascular resistance (SVR) decreased, but portal venous inflow (PVI) markedly increased (P<0.05). Seven or 15 days of L-NNA treatment reduced COX-1 mRNA expression in these vessels and the small intestine, concomitant with a significant decrease in the concentration of plasma PGI(2) and serum NO in IHPH and PHPH rats (P<0.05). At the same time, PVI decreased but SVR increased significantly (P<0.05). In both IHPH and PHPH rats, the COX-1 mRNA expression and the concentration of plasma PGI(2) after No synthase (NOS) blockade for 15 days were higher than those for 7 days, whereas the hyperdynamic circulatory state was improved after NOS blockade for 15 days compared with 7 days. The concentration of PGI(2) treated by INDO for 15 days was not significantly different from that after 7-day COX blockade, and hemodynamics restored hyperdynamic circulatory state. CONCLUSIONS: The hyperdynamic circulatory state in rats with PHT is correlated with the concentration of serum NO. There is a possible interaction between PGI(2) and NO in the hyperhemodynamics of PHT. PGI(2) is probably not the mediator in the formation and development of the hyperdynamic circulatory state in rats with chronic PHT. PMID- 17085344 TI - Feasibility and effectiveness of chemical bile duct embolization for chemical hepatectomy: a preliminary study. AB - BACKGROUND: The high operative risk of hepatectomy for specially located intrahepatic stones is still a problem to be solved. This study was undertaken to investigate the feasibility and effectiveness of chemical bile duct embolization for chemical hepatectomy. METHODS: Oxybenzene or absolute ethanol plus N-butyl cyanoacrylate was employed for embolization. The feasibility, effectiveness and mechanism of chemical hepatectomy were preliminarily analyzed histologically or by Fas, TIMP-1, TGF-beta(1), and collagen I. RESULTS: Oxybenzene plus cyanonacrylate can preferably destroy and embolize the intrahepatic biliary duct, leading to the disappearance of hepatocytes in the periphery of embolized lobe and the achievement of effective chemical hepatectomy. The expressions of Fas, TIMP-1 and TGF-beta(1) in oxybenzene embolism group (88.90 +/- 38.10, 619.43 +/- 183.42, 185.22 +/- 70.39) and ethanol embolism group (72.39 +/- 29.51, 407.55 +/- 134.74, 163.56 +/- 51.75) were higher than those of biliary duct-ligated group (26.31 +/- 12.07, 195.31 +/- 107.67, 74.84 +/- 40.73) (P<0.05). The collagen I positive percentage in the oxybenzene embolism group was also greater than that of the ethanol embolism group (33.97 +/- 12.51% vs. 20.67 +/- 8.09%, P<0.05). CONCLUSION: The effect of chemical hepatectomy may be achieved by chemical bile duct embolization. PMID- 17085345 TI - Melatonin abates liver ischemia/reperfusion injury by improving the balance between nitric oxide and endothelin. AB - BACKGROUND: Melatonin exerts complex physiological and pharmacological effects on multiple systems and organs. We hypothesized that melatonin might abate ischemia/reperfusion (I/R) injury in the liver by inhibiting excessive oxidative stress and keeping nitric oxide (NO) from being scavenged by free radicals. The aim of the present study was to investigate whether melatonin protects the liver from I/R injury and, if so, by what underlying mechanism. METHODS: Under anesthesia, Wistar rats were intraperitoneally injected with 20 mg/kg melatonin (dissolved in physiological saline containing 4% ethanol, Mel group), 4% alcohol (Alc group), or physiological saline (NS group). The artery, portal vein and bile duct of the left lobe of the liver were clamped for 60 minutes and then released. At different time points after I/R, the rats were sacrificed and blood samples were collected to measure the levels of serum alanine aminotransferase (ALT), lactic dehydrogenase (LDH), and NO. Hepatic tissue samples were collected for measuring endothelin expression by immunohistochemical staining and for routine morphological and histological examination. RESULTS: The levels of both ALT and LDH in the Mel group were significantly reduced for up to 24 hours after I/R compared with the Alc and NS groups (P<0.05). The levels of NO in the Mel group were significantly elevated for up to 12 hours after I/R relative to the NS group (P<0.05). The NO levels were also elevated at 0.5 and 6 hours after I/R in the Alc group (P<0.05). The immunohistochemical staining of hepatic tissue showed that endothelin-positive cells were significantly fewer in the Mel group than in the Alc and NS groups at 6 hours after I/R (P<0.01). The necrosis of hepatocytes and the destruction of hepatic cords in the Alc and NS groups were greatly improved in Mel-treated rats, which is in concert with our functional data. CONCLUSIONS: Pretreatment with melatonin increased NO bioavailability and decreased endothelin expression, and consequently played a protective role in preserving both liver function and structure during ischemia and reperfusion injury. PMID- 17085346 TI - Relationship between expression of Smac and Survivin and apoptosis of primary hepatocellular carcinoma. AB - BACKGROUND: The second mitochondria-derived activator of caspase/direct inhibitor of apoptosis-binding protein with low pI (Smac/DIABLO) was recently identified as a protein that is released from mitochondria in response to apoptotic stimuli and promotes apoptosis by antagonizing inhibitor of apoptosis proteins. Furthermore, Smac/DIABLO plays an important regulatory role in the sensitization of cancer cells to both immune- and drug-induced apoptosis. However, little is known about the clinical significance of Smac/DIABLO in various cancers including hepatocellular carcinoma (HCC). This study was undertaken to investigate the expression of Smac and Survivin and their relationship with the apoptosis in primary HCC. METHODS: The expression of Smac and Survivin proteins was evaluated by immunohistochemistry. The mRNA expression of Smac and Survivin was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) in HCC tissues of 50 patients, para-carcinoma tissues of 20 patients, and normal liver tissues of 15 patients. RESULTS: Smac mRNA was detected by RT-PCR in HCC tissues of 21 (42.0%) of the 50 patients, para-carcinoma tissues of 19 (95.0%) of the 20 patients, and normal liver tissues of 15 (100%) of the 15 patients. Survivin mRNA was found in HCC tissues of 46 of the 50 patients, para-carcinoma tissues of 2 of the 20 patients, and normal liver tissues of 0 of 15 patients. Immunohistochemistry revealed Smac protein in HCC tissues of 20 patients (40.0%), in para-carcinoma tissues of 18 patients (90.0%), and normal liver tissues of 15 patients (100.0%). The expression of Smac was significantly different in HCC tissues and non-HCC tissues. Survivin protein was found in HCC tissues in 45 patients, para-carcinoma tissues in 2 patients, and normal liver tissues in none of the patients. The expression of Survivin was significantly different in HCC tissues and non-HCC tissues. CONCLUSION: Smac inhibits apoptosis of HCC cells by suppression of Survivin, and the two genes probably form an important link in the signal pathway of HCC cells. PMID- 17085347 TI - Role of laparoscopic subtotal cholecystectomy in the treatment of complicated cholecystitis. AB - BACKGROUND: Laparoscopic cholecystectomy (LC) has become the "gold standard" in treating benign gallbladder diseases. Increasing laparoscopic experience and techniques have made laparoscopic subtotal cholecystectomy (LSC) a feasible option in more complex procedures. In recent years, few studies with a few cases of LSC have reported good results in patients with various types of cholecystitis. This study was designed to evaluate the feasibility, indications, characteristics and benefits of LSC in patients with complicated cholecystitis. METHODS: Altogether, 3485 patients were scheduled to receive LC during the past 4 years at our institute. Among them, 168 patients with various complicated forms of cholecystitis were treated by LSC. Meanwhile, the other 3317 patients who received standard LC were enrolled as the control group. Perioperative data from the two groups were collected and retrospectively analyzed. RESULTS: In the LSC group, 135 patients suffered from acute calculic cholecystitis, 18 from chronic calculic cholecystitis with cirrhotic portal hypertention, and 15 from chronic calculic atrophy cholecystitis with severe fibrosis. These patients constituted 4.8% of the total patients who underwent LC (168/3485) in the same period at our institute. In 122 patients, the cystic duct and artery were clipped before division. In another 46 patients, the gallbladder was initially incised at Hartmann's pouch. Five patients (3.0%) were converted to open subtotal cholecystectomy. The median operation time for LSC was 65.5 +/- 15.2 minutes, estimated operative blood loss was 71.5+/-15.5 ml, and the time to resume diet was 20.4 +/- 6.3 hours. Thirteen patients (7.7%) had local complications. The mean postoperative hospital stay was 4.2 +/- 2.6 days. In the LC group, 2887 had chronic calculic cholecystitis, 312 had acute calculic cholecystitis, 47 had chronic calculic atrophy cholecystitis, and 71 had polypus. Seventeen patients (0.5%) were converted to open cholecystectomy. The median operation time was 32.6 +/- 10.2 minutes, the estimated operative blood loss was 24.5 +/- 8.5 ml, and the time to resume diet was 18.3 +/- 4.5 hours. Thirty-nine patients (1.2%) had local complications. Mean postoperative hospital stay was 3.8 +/- 1.4 days. There was no bile duct injury or mortality in either group. CONCLUSIONS: LSC for patients with complicated cholecystitis is difficult, with a longer operation time, more operative blood loss and higher conversion and complication rates than LC. However, it is feasible and relatively safe. LSC is advantageous over open surgery, but it remains a non-routine choice. It is important to know the technical characteristics of LSC, and pay attention to perioperative bleeding and bile leak. PMID- 17085348 TI - Application of needle-knife in difficult biliary cannulation for endoscopic retrograde cholangiopancreatography. AB - BACKGROUND: Getting directly into the common bile duct (CBD) is the most important step for successful therapeutic biliary endoscopy. In 5%-10% of cases, the CBD remains inaccessible, necessitating pre-cut papillotomy or fistulotomy with a needle-knife. The aim of this study was to assess the value of early application of the needle-knife in difficult biliary cannulation for endoscopic retrograde cholangiopancreatography (ERCP). METHODS: Patients with failed biliary cannulation after 10 minutes or guide wire entering the pancreatic tube 3 times were randomly divided into group of needle-knife cut and group of persistent cannulation by standard techniques. The cannulation times, success rates and complication rates were compared between the two groups. RESULTS: A total of 948 therapeutic biliary ERCP procedures were performed between October 2004 and February 2006. Of 91 patients with difficult biliary cannulation, 43 patients underwent needle-knife cut: the cannulation success rate was 90.7%, the mean cannulation time was 5.6 minutes, and the complication rate was 9.3%. The other 48 patients underwent persistent cannulation by standard techniques: the cannulation success rate was 75%, the mean cannulation time was 10.2 minutes, and the complication rate was 14.6%. Significant differences were observed in cannulation success rate and cannulation time but in complication rate between the two groups. CONCLUSION: The early application of the needle-knife in difficult biliary cannulation is time-saving, safe and effective, with no increase in complication rate. PMID- 17085349 TI - 103Pd radioactive stent inhibits biliary duct restenosis and reduces smooth muscle actin expression during duct healing in dogs. AB - BACKGROUND: This study was designed to assess the expression of smooth muscle actin (SMA) in the healing process after implanting a (103)Pd radioactive stent in the biliary duct, and to discuss the function and significance of this stent in preventing biliary stricture formation. METHODS: A model of biliary injury in dogs was made and then a (103)Pd radioactive stent was positioned in the biliary duct. The expression and distribution of SMA were assessed in the anastomotic tissue 30 days after implantation of the stent. RESULTS: SMA expression was less in the (103)Pd stent group than in the ordinary stent group. The (103)Pd stent inhibited scar contracture and anastomotic stenosis. CONCLUSION: The (103)Pd stent can reduce the expression of SMA in the healing process and inhibit scar contracture and anastomotic stenosis in the dog biliary duct. PMID- 17085350 TI - Outcome of severe acute pancreatitis: is there a role for conservative management of infected pancreatic necrosis? AB - BACKGROUND: Infected pancreatic necrosis is associated with high morbidity and mortality and is mandatory for surgical or radiological intervention. A selected group of patients with CT evidence of infected pancreatic necrosis and a comparatively lower APACHE score may be clinically stable throughout the course of their illness. METHODS: Case records of 52 patients with severe acute pancreatitis admitted from October 2000 to September 2005 were retrospectively analysed to assess the feasibility of conservative management of infected pancreatic necrosis. CT evidence of retroperitoneal air pockets, deteriorated clinical condition, sepsis and positive blood culture were used to diagnose infected pancreatic necrosis. RESULTS: In the 52 male patients reviewed, 24 patients had infected pancreatic necrosis. Eighteen patients who had progressively deteriorated clinical conditions required surgical intervention; five patients of whom (27.8%) died. Six patients with transient end organ dysfunction and stable clinical conditions were treated with prolonged administration of antibiotics and ICU support. All these patients recovered and discharged from the hospital, and no symptoms or readmission happened during follow-up of 6-44 months. CONCLUSIONS: Selected patients with infected pancreatic necrosis who are clinically stable with transient end organ dysfunction can be treated conservatively with a favourable outcome. Necrosectomy associated with high morbidity and mortality in these patients can be avoided. The need for intervention should be individualized and based on clinical conditions of the patients. PMID- 17085351 TI - Ligustrazine alleviates acute lung injury in a rat model of acute necrotizing pancreatitis. AB - BACKGROUND: Acute necrotizing pancreatitis leads to a systemic inflammatory response characterized by widespread leukocyte activation and, as a consequence, distant lung injury. The aim of this study was to evaluate the effect of ligustrazine, extracted from Ligusticum wallichii a traditional Chinese medicine, on lung injury in a rat model of acute necrotizing pancreatitis (ANP). METHODS: A total of 192 rats were randomly divided into three groups: control (C group); ANP without treatment (P group); and ANP treated with ligustrazine (T group). Each group was further divided into 0.5, 2, 6 and 12 hours subgroups. All rats were anesthetized with an intraperitoneal injection of sodium pentobarbital. Sodium taurocholate was infused through the pancreatic membrane to induce ANP. For the T group, sodium taurocholate was infused as above, then 0.6% ligustrazine was administered via the femoral vein. The effects of ligustrazine on the severity of lung injury were assessed by lung wet/dry weight ratio, myeloperoxidase (MPO) activity and histopathological changes. Pulmonary blood flow was determined by the radioactive microsphere technique (RMT). RESULTS: The blood flow in the P group was significantly lower than that of the C group, while the blood flow in the T group was significantly higher than that of the P group but showed no significant difference from the C group. Compared with C group, the lung wet/dry ratios in both the P and T groups were significantly increased, but there was no significant difference between them. The MPO activity in the P group was greatly increased over that of the C group. In the T group, although the MPO activity was also higher than in the C group, it much less increased than in the P group. Moreover, the difference between P and T groups was significant after 0.5 to 12 hours. After induction of the ANP model, the pancreas showed mild edema and congestion; the longer the time, the more severe this became. The pulmonary pathological changes were aggravated significantly in the P group. Histopathological scores were higher in the P group than in the C group throughout the experimental course. Histopathological scores in the T group were lower than those in the P group at 6 and 12 hours. CONCLUSIONS: Microcirculatory disorder is an important factor of lung injury in ANP. Ligustrazine can ameliorate microcirculatory disorder and alleviate the damage to the lung. PMID- 17085352 TI - Surgical resection of a solitary liver metastasis from nasopharyngeal carcinoma: a case report. AB - BACKGROUND: Nasopharyngeal carcinoma (NPC) has a propensity to develop distant metastases at a high rate and with poor prognosis. Metastatic sites are usually multifocal and involve bones, lungs, liver and distant lymph nodes. Management of metastatic disease is essentially palliative and is based on chemotherapy. METHODS: A 50-year-old man with a solitary liver metastasis from a newly diagnosed NPC was treated by segmentectomy. Prior to surgery, neoadjuvant chemotherapy followed by concurrent chemoradiotherapy was administered. RESULTS: Complete remission of the primary disease was achieved, although the size of the hepatic lesion was increased. After resection of the liver metastasis, no signs of local or distant recurrence was noted during the 6-month follow up. CONCLUSION: Although surgical treatment has a limited role in metastatic NPC, there are rare cases of localized disease with a reasonable outcome after resection. PMID- 17085353 TI - Non-composite combined liver and intestinal allotransplantation. AB - BACKGROUND: Patients with short bowel syndrome may require combined liver and intestinal transplantation due to total parenteral nutrition(TPN)-related liver damage. We report combined liver and intestinal allotransplantation as a non composite technique in a patient in China. METHODS: During the operation, a 380 cm long intestine was transplanted with systemic drainage and aortic inflow, while the liver graft was placed in a piggyback fashion. Warm ischemic time of the donor graft was 2 minutes and 30 seconds, and cold ischemic time for intestinal and the liver graft was 6 hours and 40 minutes and 8 hours and 7 minutes, respectively. Immunosuppressants used after operation included tacrolimus, methylprednisolone, mycophenolate mofetil and Zenapax. RESULTS: The recipient recovered with no evidence of rejection and was kept well on tube feeding. Eventually, he died of massive hemorrhage of the thoracic cavity on day 210 after transplantation. CONCLUSION: The non-composite combined liver and intestinal allotransplantation is superior to composite technique in adult patients, particularly those who have had abdominal infection or repeated abdominal operations. PMID- 17085354 TI - Dubin-Johnson syndrome with systemic lupus erythematosus: a case report. AB - BACKGROUND: Dubin-Johnson syndrome (DJS) is a rare clinical entity. We describe a case of DJS complicated by systemic lupus erythematosus (SLE). METHODS: A case of congenital hyperbilirubinemia with SLE was evaluated systematically including review of history, physical examination for the stigmata of chronic liver disease, and other investigations. RESULT: Liver biopsy revealed a black liver with preserved architecture suggestive of DJS. CONCLUSIONS: SLE may develop in DJS. The relationship between DJS and SLE in this case is most likely a chance occurrence. PMID- 17085355 TI - Primary hepatic malignant fibrous histiocytoma mimicking cystadenocarcinoma: a case report. AB - BACKGROUND: Malignant fibrous histiocytoma is the most common sarcoma of soft tissue, which occurs usually in the extremities, but less common in the retroperitoneal space, abdominal cavity or other sites. Primary malignant fibrous histiocytoma of the liver is extremely rare; only 28 cases have been reported to date in the English literature. METHODS: In this report, a case of primary malignant fibrous histiocytoma of the liver was described in terms of clinical presentations, diagnosis and treatment and outcome. RESULTS: A 50-year-old man had a large multicystic-mass lesion in the left lobe of the liver, which was inoperable by laparotomy. Pathological examination of biopsy specimen after operation confirmed a malignant fibrous histiocytoma of storiform-pleomorphic type. The tumor developed rapidly, and the patient died of hepatic failure 2 months after the surgery. CONCLUSIONS: Primary malignant fibrous histiocytoma of the liver is diagnosed in late stage because of its rarity and non-specific presentations. Surgical resection, if feasible, is the first step treatment. The prognosis of primary malignant fibrous histiocytoma of the liver is grim with a median survival of 3 months as reported. Surgeons should be alert to the existence of this type of soft tissue tumor in the liver. PMID- 17085356 TI - Combined use of choledochoscope and duodenoscope in treatment of bile peritonitis after removal of T-tube. AB - BACKGROUND: Biliary leakage after removal of a T-tube has significant morbidity and mortality. Its etiology is multifactorial. The treatment and outcome of this complication vary. In the present study we evaluated the procedures and efficacy of combined use of choledochoscope and duodenoscope in the treatment of bile peritonitis after T-tube removal. METHODS: The procedures and results of 11 cases of biliary leakage after removal of T-tube who had been treated from January 1998 to June 2004 by combined use of choledochoscope and duodenoscope were analyzed retrospectively. RESULT: After the treatment, 9 patients were cured, and 2 were reoperated on and cured. CONCLUSIONS: Biliary leakage after removal of T-tube can be cured successfully by combined use of choledochoscope and duodenoscope. Importantly, the method is simple, effective and safe, and mostly reoperation can be avoided. PMID- 17085358 TI - The cerebrovascular disease basis of psychiatric syndromes: a model for the power of disease reasoning in psychiatry. PMID- 17085359 TI - Compendium of cerebrovascular diseases. AB - Stroke is characterized by the sudden loss of circulation to an area of the brain, resulting in a corresponding loss of neurological function. In this article, we will discuss the epidemiology, etiopathophysiology, and symptomatology of different types of cerebrovascular diseases. Specific stroke syndromes will be reviewed to convey the myriad presentations of cerebrovascular disease. PMID- 17085360 TI - Methodology for measuring cerebrovascular disease burden. AB - Biological aging of the brain is partly attributable to aging of the cerebrovascular circulation and the effects of these vascular changes on the brain. A variety of techniques ranging from simple, clinical scores to complex radiological techniques have been used in an attempt to understand, describe and quantify this process. Simultaneously attempts have been made to relate these changes to cognitive and physical changes and the risk of dementia and stroke associated with brain aging. The most frequently used clinical scores are the Framingham Stroke Risk Profile and the Hachinski Ischemic Score for vascular dementia. Radiological techniques to estimate cerebrovascular burden include many varieties of ultrasonographic, computerized tomography (CT), magnetic resonance imaging (MRI) and nuclear medicine techniques. The radiological techniques evaluate the nature and extent of disease in the vessels supplying the brain and the pattern and extent of radiological evidence of damage to the brain both on static and dynamic imaging and are briefly outlined in this review. There are several studies using these techniques to study 'normal' aging populations, and the techniques used in the most widely known of these studies are briefly highlighted in this review. PMID- 17085361 TI - Cerebrovascular disease and dementia. AB - Cerebrovascular disease (CVD) is an important cause of psychiatric disability in the elderly. Much of this disability can be attributed to dementia and lesser degrees of cognitive impairment, which result from strokes and other forms of cerebrovascular pathology. While vascular dementia is common, estimates of its frequency vary due to its clinical and pathologic heterogeneity, the challenges involved in its measurement and its frequent co-occurrence with Alzheimer's disease. Nevertheless the clinical features and natural histories of vascular dementia can be described, and risk factors have been identified and include hypertension, diabetes mellitus, hyperlipidaemia, other conditions that promote atherosclerosis, and rare genetic mutations. While vascular dementia is not curable, treatments are available. For example, a few recent clinical trials suggest that cholinesterase inhibitors have some efficacy. Our knowledge of the risk factors has also provided opportunities for the primary and secondary prevention of vascular dementia, and indicates promising avenues for research. PMID- 17085362 TI - Cerebrovascular disease basis of depression: post-stroke depression and vascular depression. AB - The close association between cerebrovascular disease and depression has been known for more than a century, yet much of the progress in understanding the cerebrovascular basis of depression in late life has been spurred by development of two concepts: 'post-stroke depression' and 'vascular depression'. The purpose of this review is to examine the epidemiology, diagnostic features, course, pathophysiology and prognosis of post-stroke depression and vascular depression, to highlight their common features, and to contrast the distinct aspects of these two subtypes of geriatric depression. PMID- 17085363 TI - Neuroimaging in late-life depression. AB - Late-life depression may be associated with vasculopathy. Neuroimaging has been a critical tool in exploring the relationship between this form of depression and vascular factors. Magnetic resonance imaging has been the most widely used tool, but there is potential to use other structural imaging techniques as well as functional neuroimaging methodologies. Neuroimaging may potentially be utilized at some point as a biomarker for late-life depression, thus helping with diagnosis and guiding treatment. PMID- 17085364 TI - Treatment of psychiatric syndromes due to cerebrovascular disease. AB - Neuropsychiatric syndromes are common in the setting of cerebrovascular disease. The most frequent psychiatric syndrome after stroke is depression. Emotionalism and apathy after stroke are also frequent and under-detected symptoms. Treatment principles are broadly similar to those currently used to treat non-organically ill patients. The evidence for pharmacological and psychological treatment for depression after stroke is scant, and of variable quality. Currently there is evidence of efficacy for both tricyclic antidepressants and SSRIs in the management of depression but the latter are better tolerated. Randomized controlled trials of antidepressants for post-stroke emotionalism are positive and this is encouraging. The current evidence base for psychological interventions either as first line or augmentative strategies is too limited and inconclusive to permit definite recommendations. Future studies might include multi-modal interventions using the principles of active case management and pharmacological studies which target both specific neuropsychiatric symptoms and underlying cerebrovascular disorder. PMID- 17085365 TI - Preventing depression after stroke. AB - Mental health prevention research is an emerging and intriguing field. Preventing mental disorders is attractive as successful prevention, as it could potentially avert both emotional suffering and illness related morbidity and mortality. Several studies have looked at preventing post-stroke depression, a common complication of stroke. In this review article, we will first provide a conceptual overview of mental health prevention research. Then, we will discuss recent research supporting the prevention of depression after stroke and the likely positive effect successful prevention may have on non-psychiatric outcomes. PMID- 17085371 TI - Not all placebos are equally pleasing. PMID- 17085372 TI - Is HRT justified for symptom management in women at higher risk of developing breast cancer? AB - Hormone replacement therapy (HRT) is the most efficacious intervention for the treatment of estrogen-deficiency symptoms. Prescriptions for HRT have fallen over the last 3 years due to anxiety provoked about breast cancer risk and recurrence that has been generated by recent clinical trials. In women at population risk of breast cancer, these trials have not shown risks greater than estimates from clinical trial evidence that predated them. For women at increased breast cancer risk due to a family history or high-risk benign breast conditions, clinical trial data are limited but suggest a lack of an additive effect of HRT on risk. In symptomatic breast cancer survivors, observational data suggest no increase in recurrence but these data are open to bias. Interim analyses of large, randomized trials have shown contradictory outcomes and, as a result, three large HRT randomized trials have now been closed. The randomized LIBERATE trial evaluating tibolone in breast cancer survivors is fully recruited and continuing. The current clinical climate is 'HRT adverse' but, due to a lack of effective alternatives for symptom relief, women at higher breast cancer risk and breast cancer survivors are still requesting information about HRT. In this situation, discussion of the current clinical uncertainty surrounding the use of HRT must be undertaken to ensure that women are adequately informed. PMID- 17085373 TI - Risk of cardiovascular disease in relation to the use of combined postmenopausal hormone therapy: detection bias and resolution of discrepant findings in two Women's Health Initiative studies. AB - CONTEXT: In a recent analysis of data in the Women's Health Initiative (WHI) clinical trial and observational studies, the overall risks of coronary heart disease, stroke and venous thromboembolism, in postmenopausal women who used combined therapy with conjugated estrogens and medroxyprogesterone acetate, were lower in the observational data. However, the risks became more similar within similar duration categories of follow-up. In both studies, there was initial elevation, followed by a duration-dependent reduction, in the risks. The investigators suggest that the discrepancies in the overall risks may be due to weighting by short-duration exposure in the clinical trial, and long-duration exposure in the observational study. CRITIQUE: In the clinical trial, 44% of the hormone therapy recipients were unblinded, mainly because of persistent vaginal bleeding, and awareness of exposure status was therefore common; in the observational study, all exposed women were aware. When follow-up in the WHI studies commenced, extensive publicity was given to evidence to suggest that, among women who had already sustained a myocardial infarction, combined hormone therapy may not decrease the risk of a second myocardial infarct, as hypothesized, but instead increase it. From the commencement of follow-up onward, detection bias was therefore possible in both WHI studies. That possibility was further reinforced after about 3 years of follow-up, and again a year later, when the clinical trial participants were explicitly informed that supplemental hormones were associated with increased cardiovascular risks. That information was also given extensive publicity. CONCLUSIONS: Detection bias could not be ruled out in either WHI study, and there may have been systematic overestimation of the risks of cardiovascular outcomes associated with the use of combined hormone therapy. To a limited extent, an analysis of the clinical trial data according to blinding status might be informative in determining whether detection bias could have given rise to overestimation of the overall and duration-specific risks in both WHI studies. If numbers permit, comparisons within strata of severity might also be informative. If the risks were overestimated in the WHI studies, a duration-related protective effect of combined hormone therapy on the risk of coronary heart disease and stroke may have gone undetected. PMID- 17085374 TI - Menopause in Egypt: past and present perspectives. AB - The mean age of the menopause in Egypt is 46.7 years, which is low compared to many countries, but this age has been rising recently. The incidence of menopause associated symptoms in Egyptian women is higher than in the West, probably because of the different 'sociocultural attitudes' towards the menopause in different communities. Bone mineral density charts have been constructed for Egyptian women and show that, in general, they have a lower bone mineral density compared to their Western counterparts. After the menopause, they suffer from osteoporosis, particularly at the femoral neck. Egyptian women do not know much about the menopause, except that the incidence of osteoporosis is increased. Their attitude towards the menopause is generally positive and about one-third of them regard the menopause as 'a normal physiological change'. Nevertheless, there exists a need for an awareness campaign in order to educate them about this important stage of their lives. PMID- 17085375 TI - Comparison between phytoestrogens and estradiol in the prevention of atheroma in ovariectomized cholesterol-fed rabbits. AB - OBJECTIVES: There is increasing interest in the role of complementary and alternative medicines for the treatment of menopause-related problems. This study compared the preventive effect on atheroma formation of a commercially available mixed phytoestrogen concentrate with that of estradiol. METHODS: An ovariectomized cholesterol-fed rabbit model of atheroma formation was used. Rabbits were ovariectomized before the commencement of the 12-week treatment period. There were two control groups. Control Group 1 received isoflavone-free rabbit chow whilst Control Group 2 received 1% cholesterol-enriched isoflavone free rabbit chow. Rabbits in Group 3 received 1% cholesterol-enriched isoflavone free rabbit chow plus a 500 mg tablet containing a concentrated extract of Trifolium pretense (red clover). Rabbits in Group 4 received 1% cholesterol enriched isoflavone-free rabbit chow plus a 0.5 mg tablet of oral estradiol. Atheroma formation was measured by, first, calculation of the area of atheroma on the intimal surface, and, second, measuring the cholesterol content in the aorta. RESULTS: There were no significant differences in serum cholesterol between the cholesterol-fed control Group 2 and the treatment Groups 3 and 4. However, there was significantly less staining for atheroma and significantly less cholesterol accumulation in the aorta in Group 4 (estradiol-treated) rabbits compared with either control Group 2 or Group 3 (phytoestrogen-treated) rabbits. CONCLUSION: In this study, only estradiol was shown to have a significant protective effect against atheroma formation. PMID- 17085376 TI - The long-term effects of low-dose 17beta-estradiol and dydrogesterone hormone replacement therapy on 24-h ambulatory blood pressure in hypertensive postmenopausal women: a 1-year randomized, prospective study. AB - OBJECTIVE: The aim of this study was to assess the long-term effects of low-dose oral hormone replacement therapy (HRT) on 24-h blood pressure in hypertensive postmenopausal women. STUDY DESIGN: In this 12-month, prospective study, 66 postmenopausal women with mild or moderate hypertension were randomly assigned to receive either HRT with 1 mg/day micronized 17beta-estradiol sequentially combined with 10 mg/day dydrogesterone for 14 days of each 28-day cycle, or no therapy. Ambulatory blood pressure measurements were recorded for a 24-h period at baseline and after 12 months of treatment or follow-up. RESULTS: Blood pressure did not differ significantly between the groups at baseline. After 12 months, there were falls in 24-h systolic, diastolic and mean arterial blood pressure in both the HRT and control groups; only the fall in mean arterial blood pressure in the HRT group achieved statistical significance (-2.0 +/- 0.8 mmHg, p < 0.01). While there was no significant decrease in daytime systolic or mean arterial blood pressure in either group, a significant decrease in diastolic blood pressure (-1.8 +/- 10 mmHg, p < 0.001) was observed in the HRT group. Night time systolic and mean arterial blood pressure also decreased significantly (p < 0.001) in the HRT group (-3.0 +/- 1.5 mmHg and -2.2 +/- 0.6 mmHg, respectively), but no significant change was observed in the control group. CONCLUSION: Low-dose oral HRT caused significant falls in both daytime and night-time ambulatory blood pressure in postmenopausal women with mild or moderate hypertension. PMID- 17085377 TI - Quantitative ultrasonometry of the phalanges during pregnancy: a longitudinal study. AB - OBJECTIVE: The aim of this study was to determine prospectively the change in results of bone ultrasonometry measurement during pregnancy in healthy German women. Study design Quantitative ultrasonometry (QUS) of the phalanges was performed in 60 healthy, pregnant women. Measurements of amplitude-dependent bone propagation velocity (speed of sound; AdSOS) and the bone transmission time (BTT) were performed during the three trimesters of pregnancy in 60 patients. RESULTS: During pregnancy, a significant increase in body weight and body mass index (p < or = 0.001) were observed. In accordance with bone ultrasonometry, a significant reduction in AdSOS was found in each trimester; AdSOS was significantly lower in the second and third trimesters compared with the first (p < or = 0.001). The BTT values also decreased significantly in the second and third trimesters compared with the first (p < or = 0.001). No significant influence was found of possible risk factors such as family risk of osteoporosis, previous pregnancies, age at menarche and prior use of oral contraceptives on QUS measurement results. CONCLUSIONS: During normal pregnancy, we found a significant reduction of quantitative ultrasonometry variables AdSOS and BTT in healthy pregnant women. This decrease had a large influence on the t score and Z score of QUS in our study and demonstrates therefore a possible clinical relevance. The decrease was independent of osteoporosis-related risk factors and the increase in body weight. More large-scale, prospective studies are needed to increase our knowledge about the mechanism of bone turnover during pregnancy and lactation. PMID- 17085378 TI - Vasoactive biomarkers and oxidative stress in healthy recently postmenopausal women treated with hormone replacement therapy. AB - BACKGROUND: Despite biologically plausible mechanisms for cardiac protection from estrogen therapy, recent clinical trials have suggested possible cardiovascular risk rather than benefit. However, it has been speculated that cardioprotective benefits from hormone replacement therapy (HRT) may be more evident in the early postmenopausal period. We have previously reported early beneficial effects on biochemical markers of endothelial function in healthy women after short-term estradiol replacement therapy. In this study we aimed to evaluate the effect of long-term HRT on different vasoactive factors and oxidative stress in healthy recently postmenopausal women. METHODS: Fifteen women (age 50 +/- 1 years, time since menopause 1.6 +/- 0.1 years) were randomized to a sequential oral and transdermal estradiol regimen (2 mg oral micronized 17beta-estradiol/day or 1.5 mg 17beta-estradiol gel/day). Oral dydrogesterone (10 mg/day, 12 days/month) was then cyclically combined with either of the estrogen therapies for 1 year. Blood samples were collected at baseline and after 1, 2, 6 and 12 months of therapy to evaluate levels of follicle stimulating hormone (FSH), estradiol, 6-keto PGF1alpha (prostacyclin metabolite), nitrite/nitrate, epinephrine, norepinephrine, 8-isoprostane (8-epi PGF2alpha) and lipid profile values. RESULTS: FSH levels decreased (p < 0.001) while estradiol levels increased (p < 0.001) during HRT. Levels of epinephrine (p < 0.001), norepinephrine (p < 0.01), mean blood pressure (p < 0.01) and low density lipoprotein (LDL) cholesterol (p < 0.01) decreased, and nitrite/nitrate levels increased (p < 0.01) during HRT, which did not significantly affect 8-epi PGF2alpha levels. CONCLUSIONS: One-year HRT significantly reduced the levels of catecholamines, mean blood pressure and LDL cholesterol while it increased levels of nitrite/nitrate, indicating cardiovascular benefit in healthy recent postmenopausal women. Levels of 8-epi PGF2alpha did not change, suggesting no evident relationship between HRT and oxidative stress. PMID- 17085379 TI - A study of women on long-term hormone replacement therapy and their attitude to suggested cessation. AB - OBJECTIVE: The study was undertaken to determine the effect of advice to discontinue hormone replacement therapy (HRT) on 100 women who were well established on treatment without side-effects. METHOD: The study was retrospective from November 2003 to April 2004, in a single gynecological practice in London, UK. One hundred consecutive long-term estrogen and testosterone hormone implant users were assessed as to their knowledge of recent studies regarding risks of long-term HRT and whether they wished to discontinue hormones. RESULTS: All women receiving estrogen and testosterone implants, for a mean duration of 17.65 years (range 10-28 years), felt well informed concerning the Women's Health Initiative Study and the Million Women Study but only three women were happy to discontinue. The reasons given for the continuation of therapy were that they felt well and their quality of life had greatly improved. The mean estradiol and testosterone levels were 921 pmol/l and 1.91 nmol/l, respectively. CONCLUSIONS: The high rate of continuation of hormone treatment indicates that, despite the recent adverse publicity, these women feel well informed and were not willing to discontinue with their hormone therapy if they felt well. A regular discussion of the risks and benefits of HRT remains mandatory. PMID- 17085380 TI - Effect of different preparations of hormone therapy on sexual dysfunction in naturally postmenopausal women. AB - OBJECTIVE: To compare the effect of different formulations of continuous combined hormone therapy on sexual performance in naturally postmenopausal women. MATERIAL AND METHODS: A total of 158 postmenopausal women were enrolled and prospectively randomized to the single-blind study. Fifty-four women received tibolone 2.5 mg, 53 received 2 mg estradiol and 2 mg dienogest (E2/dienogest), and 51 did not receive any menopausal therapy. The patients were monitored after 6 months. Attitudes of sexuality were evaluated by using the Rosen's female sexual function index. RESULTS: Compared with E2/dienogest and the control group, tibolone treatment was associated with more improvement of sexual performance, including sexual desire, sexual arousal and satisfaction. Both of the hormone therapies decreased frequencies of vaginal dryness and painful intercourse. CONCLUSIONS: Tibolone has more positive effects on the sexual dysfunction of postmenopausal women and may be an alternative to the E2/dienogest preparation in postmenopausal women with sexual dysfunction. PMID- 17085381 TI - ERK1/2 MAP kinases in cell survival and apoptosis. AB - ERK1/2 is an important subfamily of mitogen-activated protein kinases that control a broad range of cellular activities and physiological processes. ERK1/2 can be activated transiently or persistently by MEK1/2 and upstream MAP3Ks in conjunction with regulation and involvement of scaffolding proteins and phosphatases. Activation of ERK1/2 generally promotes cell survival; but under certain conditions, ERK1/2 can have pro-apoptotic functions. PMID- 17085382 TI - Chaperone-like activity and hydrophobicity of alpha-crystallin. AB - alpha-Crystallin, a prominent member of small heat shock protein (sHsp) family and a major structural protein of the eye lens is a large polydisperse oligomer of two isoforms, alphaA- and alphaB-crystallins. Numerous studies have demonstrated that alpha-crystallin functions like a molecular chaperone in preventing the aggregation of various proteins under a wide range of stress conditions. The molecular chaperone function of alpha-crystallin is thus considered to be vital in the maintenance of lens transparency and in cataract prevention. alpha-Crystallin selectively interacts with non-native proteins thereby preventing them from aggregation and helps maintain them in a folding competent state. It has been proposed and generally accepted that alpha crystallin suppresses the aggregation of other proteins through the interaction between hydrophobic patches on its surface and exposed hydrophobic sites of partially unfolded substrate protein. However, a quantifiable relationship between hydrophobicity and chaperone-like activity remains a matter to be concerned about. On an attentive review of studies on alpha-crystallin chaperone like activity, particularly the studies that have direct or indirect implications to hydrophobicity and chaperone-like activity, we found several instances wherein the correlation between hydrophobicity and its chaperone-like activity is paradoxical. We thus attempted to provide an overview on the role of hydrophobicity in chaperone-like activity of alpha-crystallin, the kind of evaluation done for the first time. PMID- 17085383 TI - Linking physiological mechanisms of coherent cellular behaviour with more general physical approaches towards the coherence of life. AB - Schrodinger pointed out that one of the most fundamental properties of life is its coherent behaviour. This property has been approached from a physiological point of view by Ling in his 'association-induction hypothesis' and extended by Pollack (gel-sol theory), by Chaplin and by Kaivarainen (detailed studies of cellular water). The question of coherence has also been attacked from general physics in three independent approaches: from non-linear thermodynamics (Frohlich), from quantum field theory (Del Giudice and his group) and from quantum mechanics (Davydov). In this paper all these approaches are unified. The emerging picture constitutes a new paradigm of life. PMID- 17085384 TI - Construction, expression, and characterization of a new targeted bifunctional fusion protein: tumstatin45-132-TNF. AB - The anti-angiogenic activity of tumstatin45-132 is mediated by binding to alphaVbeta3 on endothelial cells and tumor vascular endothelium showing increased expression of alphaVbeta3. Tumor necrosis factor alpha (TNF-alpha) is known to not only possess direct cytotoxicity against tumor cells, but also induces tumor vessel disruption, however, clinical use of TNF-alpha as an anticancer drug is hampered by severe systemic toxicity. In this study, we explore the possibility of fusing tumstatin45-132 with human TNF-alpha in the hope of generating a targeting, bi-functional protein in tumor treatment. Tumstatin45-132-TNF was constructed and expressed in E. coli. The recombinant fusion protein was shown to be insoluble and in an inclusion body form. An effective strategy for refolding and purification of tumstatin45-132-TNF resulted in final purified yields of 3 mg purified fusion protein recovered from 1 liter of E. coli culture. The refolded tumstatin45-132-TNF with a purity of 98% assessed by denaturing SDS - PAGE showed a single band on gels. Endothelial cell proliferation assay and standard cytolytic assays against L929 indicated that the fusion protein maintains tumstatin45-132 and TNF-alpha activity. More importantly, tumstatin45-132-TNF inhibits endothelial cell proliferation more than tumstatin45-132 alone. Cell adhesion assays and competitive binding experiments with anti-integrin antibodies showed that the tumstatin45-132 moiety specifically interacts with alphaVbeta3 integrin. These results lay the solid foundation for further investigation of antitumor activity of tumstatin45-132-TNF in vivo. PMID- 17085385 TI - Structure-based cross-linking of NF-kappaB p50 homodimer and decoy bearing a novel 2'-disulfide trapping site. AB - Double-stranded oligodeoxyribonucleotides with engineered disulfide units were successfully used for covalent trapping of cysteine containing proteins. In particular, an efficient cross-linking of NF-kappaB p50 homodimer to a sequence specific decoy was demonstrated. The results suggest that the synthetic oligonucleotides bearing a novel 2'-disulfide trapping site can be used as new tools to study and manipulate biological systems. PMID- 17085386 TI - Sequestration shapes the response of signal transduction cascades. AB - Many signal transduction cascades are composed of covalent modification cycles such as kinase/phosphatase cycles. In the 1980s Goldbeter and Koshland showed that such cycles can exhibit non-linear input-output relations when the enzymes are saturated by their substrates, which may facilitate signal processing. Recent papers show that this mechanism is unlikely to cause non-linearity in mammalian signal transduction cascades as sequestration of the target due to enzyme concentrations present in these cascades will hamper this mechanism. However, sequestration due to high-affinity enzymes can shape the dynamics and steady state behaviour of signal transduction cascades in different ways, some of which are discussed in this review. PMID- 17085387 TI - Expression of active Akt protects against tamoxifen-induced apoptosis in MCF-7 Cells. AB - We investigated the effect of constitutive active Akt expression on anti proliferative and apoptotic effect of tamoxifen in MCF-7 human breast cancer cells. Forced expression of AktDD (T308D, S473D) resulted in increased phosphorylation of GSK3beta, a physiological substrate of Akt. When estrogen receptor (ER) mediated transcription was determined by luciferase assays, there was more than 2-fold increase in estradiol-dependent transcription in MCF-7 cells overexpressing AktDD (MCF-7 AktDD) compared to vector control cells (MCF-7 vec). MCF-7 AktDD cells showed increased proliferation in a medium containing charcoal stripped serum supplemented with estradiol. When the cell cycle profiles were examined, there was an increase in S-phase and a reduction in G1 phase in MCF-7 AktDD cells as compared to MCF-7 vec cells. Overexpression of AktDD also attenuated tamoxifen-mediated apoptosis. These results suggest that Akt could confer resistance to anti-estrogen mediated cell death and inhibition of proliferation. PMID- 17085388 TI - Are the properties of mitochondrial membranes redox regulated? PMID- 17085389 TI - The coherence of life and possible mistakes in classical cell physiology: do we need a paradigm change? PMID- 17085390 TI - Challenges in the conduct of emergency research in children: a workshop report. PMID- 17085393 TI - Blastomere blasphemy. PMID- 17085394 TI - Ethics and public health emergencies: rationing vaccines. AB - There are three broad ethical issues related to handling public health emergencies. They are the three R's-rationing, restrictions and responsibilities. Recently, a severe shortage of annual influenza vaccine in the US, combined with the threat of pandemic flu, has provided an opportunity for policy makers to think about rationing in very concrete terms. Some lessons from annual flu vaccination likely will apply to pandemic vaccine distribution, but many preparatory decisions must be based on very rough estimates. What ethical principles should guide rationing decisions, what data should inform these decisions, how to revise decisions as new data emerge, and how to implement rationing decisions on the ground are all important considerations. In addition, ethicists might be able to help policy makers think through the importance of international cooperation in surmounting global rationing dilemmas and to accept the inevitable responsibilities of government in making and implementing rationing decisions. PMID- 17085395 TI - Disclosing individual genetic results to research participants. AB - Investigators and institutional review boards should integrate plans about the appropriate disclosure of individual genetic results when designing research studies. The ethical principles of beneficence, respect, reciprocity, and justice provide justification for routinely offering certain results to research participants. We propose a result-evaluation approach that assesses the expected information and the context of the study in order to decide whether results should be offered. According to this approach, the analytic validity and the clinical utility of a specific result determine whether it should be offered routinely. Different results may therefore require different decisions even within the same study. We argue that the threshold of clinical utility for disclosing a result in a research study should be lower than the threshold used for clinical use of the same result. The personal meaning of a result provides additional criteria for evaluation. Finally, the context of the study allows for a more nuanced analysis by addressing the investigators' capabilities for appropriate disclosure, participants' alternative access to the result, and their relationship with the investigators. This analysis shows that the same result may require different decisions in different contexts. PMID- 17085396 TI - Thresholds and boundaries in the disclosure of individual genetic research results. PMID- 17085397 TI - Tiered disclosure options promote the autonomy and well-being of research subjects. PMID- 17085398 TI - Best laid plans for offering results go awry. PMID- 17085399 TI - Letting the gene out of the bottle: a comment on returning individual research results to participants. PMID- 17085400 TI - When do genetic researchers have a duty to recontact study participants? PMID- 17085401 TI - Undesirable implications of disclosing individual genetic results to research participants. PMID- 17085402 TI - Disclosing genetic research results: examples from practice. PMID- 17085403 TI - Taking our obligations to research participants seriously: disclosing individual results of genetic research. PMID- 17085404 TI - Questions, complexities, and limitations in disclosing individual genetic results. PMID- 17085405 TI - Relationships with test-tubes: where's the reciprocity? PMID- 17085406 TI - Considering the nature of individual research results. PMID- 17085407 TI - One size does not fit all. PMID- 17085408 TI - Clinical utility and full disclosure of genetic results to research participants. PMID- 17085409 TI - Obligations in offering to disclose genetic research results. PMID- 17085410 TI - Ethical considerations in the communication of unexpected information with clinical implications. PMID- 17085411 TI - Analogical reasoning in handling emerging technologies: the case of umbilical cord blood biobanking. AB - How are we individually and as a society to handle new and emerging technologies? This challenging question underlies much of the bioethical debates of modern times. To address this question we need suitable conceptions of the new technology and ways of identifying its proper management and regulation. To establish conceptions and to find ways to handle emerging technologies we tend to use analogies extensively. The aim of this article is to investigate the role that analogies play or may play in the processes of understanding and managing new technology. More precisely we aim to unveil the role of analogies as analytical devices in exploring the "being" of the new technology as well the normative function of analogies in conceptualizing the characteristics and applications of new technology. Umbilical cord blood biobanking will be used as a case to investigate these roles and functions. PMID- 17085412 TI - Mixing metaphors in umbilical cord blood transplantation. PMID- 17085413 TI - Limitations and justifications for analogical reasoning. PMID- 17085414 TI - Mapping metaphors and analogies. PMID- 17085415 TI - Banking on the value of analogies in bioethics. PMID- 17085416 TI - It's not what we say, exactly...or is it? PMID- 17085417 TI - Analogical power and Aristotle's model of persuasion. PMID- 17085418 TI - Analogical trends in umbilical cord blood legislation in the United States. PMID- 17085422 TI - Special topics issue of microcirculation: ion channels and pulmonary vascular function. AB - Unique features of the pulmonary circulation impact its function in health and disease, not the least of which is the existence of developmentally distinct, functionally heterogeneous extra-alveolar and septal capillary networks. The impact of ion channel expression and regulation in lung vascular smooth muscle or endothelium in these vascular compartments provides a focus for this special topics issue. Reviews and original contributions from experts in the field discuss two broad groups of ion channels, drawing on studies utilizing biophysical and molecular approaches in heterologous expression systems, in vitro approaches in pulmonary vascular smooth muscle and endothelial cells, and physiologic studies in animal models of chronic pulmonary hypertension. First, channels involved in membrane depolarization and related alterations in vascular tone, and shear sensing or exocytosis by endothelium are discussed: voltage-gated potassium channels, ATP-regulated potassium channels and L- and T- type voltage gated calcium channels. The second series of reviews discusses the role of calcium influx pathways provided by transient receptor potential channels in regulation of pulmonary vascular tone or vascular remodeling, and endothelial barrier function. Understanding the role of ion channels in pulmonary vascular pathophysiology may be critical to development of new therapeutic strategies. PMID- 17085423 TI - The role of k+ channels in determining pulmonary vascular tone, oxygen sensing, cell proliferation, and apoptosis: implications in hypoxic pulmonary vasoconstriction and pulmonary arterial hypertension. AB - Potassium channels are tetrameric, membrane-spanning proteins that selectively conduct K+ at near diffusion-limited rates. Their remarkable ionic selectivity results from a highly-conserved K+ recognition sequence in the pore. The classical function of K+ channels is regulation of membrane potential (EM) and thence vascular tone. In pulmonary artery smooth muscle cells (PASMC), tonic K+ egress, driven by a 145/5 mM intracellular/extracellular concentration gradient, contributes to a EM of about -60 mV. It has been recently discovered that K+ channels also participate in vascular remodeling by regulating cell proliferation and apoptosis. PASMC express voltage-gated (Kv), inward rectifier (Kir), calcium sensitive (KCa), and two-pore (K2P) channels. Certain K+ channels are subject to rapid redox regulation by reactive oxygen species (ROS) derived from the PASMC's oxygen-sensor (mitochondria and/or NADPH oxidase). Acute hypoxic inhibition of ROS production inhibits Kv1.5, which depolarizes EM, opens voltage-sensitive, L type calcium channels, elevates cytosolic calcium, and initiates hypoxic pulmonary vasoconstriction (HPV). Hypoxia-inhibited K+ currents are not seen in systemic arterial SMCs. Kv expression is also transcriptionally regulated by HIF 1alpha and NFAT. Loss of PASMC Kv1.5 and Kv2.1 contributes to the pathogenesis of pulmonary arterial hypertension (PAH) by causing a sustained depolarization, which increases intracellular calcium and K+, thereby stimulating cell proliferation and inhibiting apoptosis, respectively. Restoring Kv expression (via Kv1.5 gene therapy, dichloroacetate, or anti-survivin therapy) reduces experimental PAH. Electrophysiological diversity exists within the pulmonary circulation. Resistance PASMC have a homogeneous Kv current (including an oxygen sensitive component), whereas conduit PASMC current is a Kv/KCa mosaic. This reflects regional differences in expression of channel isoforms, heterotetramers, splice variants, and regulatory subunits as well as mitochondrial diversity. In conclusion, K+ channels regulate pulmonary vascular tone and remodeling and constitute potential therapeutic targets in the regression of PAH. PMID- 17085424 TI - KATP channels are an important component of the shear-sensing mechanism in the pulmonary microvasculature. AB - OBJECTIVE: To investigate the role of a KATP channel in sensing shear, specifically its cessation, in the endothelial cells of the pulmonary microvasculature. METHODS: Endothelial cells isolated from the pulmonary microvasculature of wild-type and KATP channel knockout (KIR6.2-/-) mice were either statically cultured (non-flow-adapted) or kept under flow (flow-adapted) and the KIR currents in these cells were monitored by whole-cell patch-clamp technique during flow and its cessation. Membrane potential changes, generation of reactive oxygen species (ROS), and Ca2+ influx with flow cessation were evaluated by the use of fluorescent dyes. Lungs isolated from wild-type mice were imaged to visualize ROS generation in the subpleural endothelium. RESULTS: By patch-clamp analysis, reduction in the KIR current with cessation of flow occurred only in wild-type cells that were flow-adapted and not in flow-adapted KIR6.2-/- cells. Similar observations were made using changes in bisoxonol fluorescence as an index of cell membrane potential. Generation of ROS and Ca2+ influx that follow membrane depolarization were significantly lower in statically cultured and in KIR6.2-/- cells as compared to flow-adapted wild-type cells. Imaging of subpleural endothelial cells of the whole lung showed that the KATP antagonist glyburide caused the production of ROS in the absence of flow cessation. CONCLUSIONS: The responses to stop of flow (viz. membrane depolarization, KIR currents, ROS, Ca2+) were significantly altered with knockout of KATP channels, which indicates that this channel is an important component of the pulmonary endothelial response to abrupt loss of shear stress. PMID- 17085425 TI - T-type calcium channels in pulmonary vascular endothelium. AB - The expression of low-voltage-gated T-type calcium channel alpha1G-subunit (Cav3.1) has recently been revealed in pulmonary microvascular endothelial cells (PMVECs). In endothelial cells, changes in membrane potential may be caused by a variety of physiological or pathophysiological vascular stimuli. Thus, the characteristic low-voltage range of activation, transient kinetics of inactivation, and conducting continuous Ca2+ entry in a range of low membrane potentials of this channel may confer its temporal and spatial control of cytosolic Ca2 + important for a variety of cellular processes in vascular endothelial cells. The goal of this review is to provide a succinct description of the endothelial cell T-type calcium channel, including its molecular characteristics, biophysical and pharmacological profiles, and involvement in certain endothelial cell functions. PMID- 17085426 TI - Ca2+ channels and chronic hypoxia. AB - Many chronic lung diseases are associated with prolonged exposure to alveolar hypoxia, resulting in the development of pulmonary hypertension. While the exact mechanisms underlying the pathogenesis of hypoxic pulmonary hypertension remain poorly understood, a key role for changes in Ca2+ homeostasis has emerged. Intracellular Ca2+ concentration controls a variety of pulmonary vascular cell functions, including contraction, gene expression, growth, barrier function and synthesis of vasoactive substances. Several studies indicate that prolonged exposure to hypoxia causes alterations in the expression and activity of several Ca2+ handling pathways in pulmonary arterial smooth muscle cells. In contrast, the effect of chronic hypoxia on Ca2+ homeostasis in pulmonary arterial endothelial cells is relatively unexplored. In this review, we discuss data from our laboratory and others describing the effects of prolonged hypoxia on pulmonary vascular smooth muscle and endothelial cell Ca2+ homeostasis and the various Ca2+ channels and handling pathways involved in these responses. We will also highlight future directions of investigation that might improve our understanding of the response of pulmonary vascular cells to chronic hypoxia. PMID- 17085427 TI - TRP channels, CCE, and the pulmonary vascular smooth muscle. AB - Transient receptor potential (TRP) genes represent a novel class of genes that are generally believed to encode for nonselective cation channels. A subfamily of TRP channels, canonical TRP (TRPC), which are highly permeable to Ca2+ (and Na+), co-assembles with each other to form functional store- and receptor-operated Ca2+ channels. TRPC mRNA and protein have been identified in pulmonary arterial smooth muscle and endothelial cells. The currents generated by Ca2+ influx through store and receptor-operated Ca2+ channels have also been extensively characterized in these cells. More recently, the attention has shifted to identify the TRP subunits that underlie the function of native channels in the pulmonary vasculature, with the understanding that TRP channels assemble as either homo-or heterotetramers in vivo. This work in progress has yielded exciting information regarding the involvement of TRP channels in the control of smooth muscle contraction, and cell proliferation and migration. In this review, the authors focus on describing the function and transcriptional regulation of TRP proteins, and the store- and receptor-operated Ca2+ channels for which they are responsible, in pulmonary artery smooth muscle cells (PASMC). They also identify some key TRP proteins whose role in the pulmonary vasculature has been established, as well as some more novel subunits whose role, although intriguing, can only be inferred from other vascular studies. Finally, they describe the involvement of TRP channels in regulating pulmonary vasoconstriction, PASMC proliferation, and pulmonary endothelial barrier function. PMID- 17085428 TI - Ca2+ signaling, TRP channels, and endothelial permeability. AB - Increased endothelial permeability is the hallmark of inflammatory vascular edema. Inflammatory mediators that bind to heptahelical G protein-coupled receptors trigger increased endothelial permeability by increasing the intracellular Ca2+ concentration ([Ca2+]i). The rise in [Ca2+]i activates key signaling pathways that mediate cytoskeletal reorganization (through myosin-light chain-dependent contraction) and the disassembly of VE-cadherin at the adherens junctions. The Ca2+-dependent protein kinase C (PKC) isoform PKCalpha plays a crucial role in initiating endothelial cell contraction and disassembly of VE cadherin junctions. The increase in [Ca2+]i induced by inflammatory agonists such as thrombin and histamine is achieved by the generation of inositol 1,4,5 trisphosphate (IP3), activation of IP3-receptors, release of stored intracellular Ca2+, and Ca2+ entry through plasma membrane channels. IP3-sensitive Ca2+-store depletion activates plasma membrane cation channels (i.e., store-operated cation channels [SOCs] or Ca2+ release-activated channels [CRACs]) to cause Ca2+ influx into endothelial cells. Recent studies have identified members of Drosophila transient receptor potential (TRP) gene family of channels that encode functional SOCs in endothelial cells. These studies also suggest that the canonical TRPC homologue TRPC1 is the predominant isoform expressed in human vascular endothelial cells, and is the essential component of the SOC in this cell type. Further, evidence suggests that the inflammatory cytokine tumor necrosis factor alpha can induce the expression of TRPC1 in human vascular endothelial cells signaling via the nuclear factor-kappaB pathway. Increased expression of TRPC1 augments Ca2+ influx via SOCs and potentiates the thrombin-induced increase in permeability in human vascular endothelial cells. Deletion of the canonical TRPC homologue in mouse, TRPC4, caused impairment in store-operated Ca2+ current and Ca2+-store release-activated Ca2+ influx in aortic and lung endothelial cells. In TRPC4 knockout (TRPC4-/-) mice, acetylcholine-induced endothelium-dependent smooth muscle relaxation was drastically reduced. In addition, TRPC4-/- mouse lung endothelial cells exhibited lack of actin-stress fiber formation and cell retraction in response to thrombin activation of protease-activated receptor-1 (PAR-1) in endothelial cells. The increase in lung microvascular permeability in response to PAR-1 activation was inhibited in TRPC4-/- mice. These results indicate that endothelial TRP channels such as TRPC1 and TRPC4 play an important role in signaling agonist-induced increases in endothelial permeability. PMID- 17085429 TI - Regulation of endothelial cell barrier function by store-operated calcium entry. AB - Transient receptor potential (TRP) channels fulfill important and diverse signaling functions, and are generally conserved among species. The canonical subfamily of TRP proteins, TRPC channels, possesses 7 isoforms that combine in various ways to form heteromultimers. In endothelium, TRPC1 and TRPC4 form subunits of a channel that selectively conducts calcium. This channel is activated by calcium depletion in the endoplasmic reticulum, and thus TRPC1/TRPC4 forms the molecular basis of a store operated calcium entry pathway. TRPC4 interacts with protein 4.1, which tethers the channel to the membrane skeleton and represents a gating mechanism required for calcium permeation. In response to inflammatory agonists such as thrombin and bradykinin, the generation of inositol 1,4,5-trisphosphate transiently depletes endoplasmic reticulum calcium and activates the TRPC1/TRPC4 channel. Calcium permeation through this channel triggers cytoskeletal reorganization that is necessary to disrupt the endothelial cell barrier and increase permeability. Thus, inhibition of the TRPC1/TRPC4 channel provides a putative anti-inflammatory strategy. PMID- 17085430 TI - Ca2+ channels and pulmonary endothelial permeability: insights from study of intact lung and chronic pulmonary hypertension. AB - Phenotypic heterogeneity in pulmonary vascular endothelial cells extends to regulation of endothelial permeability, a process which often depends upon Ca2 + entry from the extracellular space. Scanning electron microscopy of vascular corrosion casts has documented distinct patterns of barrier disruption. Store depletion and activation of Ca2 + entry via canonical transient potential channels (TRPC1 and TRPC4) disrupts the barrier in extraalveolar vessels. In contrast, numerous other models of acute lung injury, including high vascular pressure- or epoxyeicosatrienoic acid-induced injury, specifically disrupt the alveolar septal barrier. This review discusses Ca2 + permeant channels which potentially could be involved in regulation of barrier integrity in the alveolar septal compartment: transient receptor potential channels, cyclic nucleotide gated channels, purinergic (P2X) channels, and T-type voltage gated channels. The evidence for the vanilloid transient receptor potential channel TRPV4 in regulating septal barrier function is discussed. Adaptations in barrier function in chronic pulmonary hypertension are reviewed, notably the loss of a store depletion-dependent permeability response in the intact lung. Finally, the authors propose that since specific disruption of the alveolar septal barrier will have deleterious functional consequences, such as alveolar flooding and impairment of gas exchange, identification of specific molecular targets for Ca2 + entry-dependent regulation of barrier function in this compartment is needed. PMID- 17085431 TI - Dividing the large glycoside hydrolase family 13 into subfamilies: towards improved functional annotations of alpha-amylase-related proteins. AB - Family GH13, also known as the alpha-amylase family, is the largest sequence based family of glycoside hydrolases and groups together a number of different enzyme activities and substrate specificities acting on alpha-glycosidic bonds. This polyspecificity results in the fact that the simple membership of this family cannot be used for the prediction of gene function based on sequence alone. In order to establish robust groups that show an improved correlation between sequence and enzymatic specificity, we have performed a large-scale analysis of 1691 family GH13 sequences by combining clustering, similarity search and phylogenetic methods. About 80% of the sequences could be reliably classified into 35 subfamilies. Most subfamilies appear monofunctional (i.e. contain enzymes with the same substrate and the same product). The close examination of the other, apparently polyspecific, subfamilies revealed that they actually group together enzymes with strongly related (or even sometimes virtually identical) activities. Overall our subfamily assignment allows to set the limits for genomic function prediction on this large family of biologically and industrially important enzymes. PMID- 17085432 TI - Role for neutral sphingomyelinase-2 in tumor necrosis factor alpha-stimulated expression of vascular cell adhesion molecule-1 (VCAM) and intercellular adhesion molecule-1 (ICAM) in lung epithelial cells: p38 MAPK is an upstream regulator of nSMase2. AB - Neutral sphingomyelinases (N-SMases) are major candidates for stress-induced ceramide production. However, there is little information on the physiological regulation and roles of the cloned N-SMase enzyme, nSMase2. In this study, nSMase2 was found to translocate acutely to the plasma membrane of A549 epithelial cells in response to tumor necrosis factor alpha (TNF-alpha) in a time and dose-dependent manner. Additionally, TNF-alpha increased N-SMase activity rapidly and transiently both endogenously and in cells overexpressing nSMase2. Furthermore, the translocation of nSMase2 was regulated by p38-alpha MAPK, but not ERK or JNK, and the increase in endogenous N-SMase activity was abrogated by p38 MAPK inhibition. In addition, both p38-alpha MAPK and nSMase2 were implicated in the TNF-alpha-stimulated up-regulation of the adhesion proteins vascular cell adhesion molecule-1 (VCAM) and intercellular adhesion molecule-1 (ICAM), but this was largely independent of NF-kappaB activation. These data reveal p38 MAPK as an upstream regulator of nSMase2 and indicate a role for nSMase2 in pro-inflammatory responses induced by TNF-alpha as a regulator of adhesion proteins. PMID- 17085433 TI - Increased functional diversity of plant K+ channels by preferential heteromerization of the shaker-like subunits AKT2 and KAT2. AB - Assembly of plant Shaker subunits as heterotetramers, increasing channel functional diversity, has been reported. Here we focus on a new interaction, between AKT2 and KAT2 subunits. The assembly as AKT2/KAT2 heterotetramers is demonstrated by (i) a strong signal in two-hybrid tests with intracytoplasmic C terminal regions, (ii) the effect of KAT2 on AKT2 subunit targeting in tobacco cells, (iii) the complete inhibition of AKT2 currents by co-expression with a dominant-negative KAT2 subunit in Xenopus oocytes, and reciprocally, and (iv) the appearance, upon co-expression of wild-type AKT2 and KAT2 subunits, of new channel functional properties that cannot be explained by the co-existence of two kinds of homotetrameric channels. In particular, the instantaneous current, characteristic of AKT2, displayed new functional features when compared with those of AKT2 homotetramers: activation by external acidification (instead of inhibition) and weak inhibition by calcium. Single channel current measurements in oocytes co-expressing AKT2 and KAT2 revealed a strong preference for incorporation of subunits into heteromultimers and a diversity of individual channels. In planta, these new channels, which may undergo specific regulations, are likely to be formed in guard cells and in the phloem, where they could participate in the control of membrane potential and potassium fluxes. PMID- 17085434 TI - Dephosphorylation of the linker regions of Smad1 and Smad2/3 by small C-terminal domain phosphatases has distinct outcomes for bone morphogenetic protein and transforming growth factor-beta pathways. AB - Smad proteins transduce bone morphogenetic protein (BMP) and transforming growth factor-beta (TGFbeta) signals upon phosphorylation of their C-terminal SXS motif by receptor kinases. The activity of Smad1 in the BMP pathway and Smad2/3 in the TGFbeta pathway is restricted by pathway cross-talk and feedback through protein kinases, including MAPK, CDK2/4, p38MAPK, JNK, and others. These kinases phosphorylate Smads 1-3 at the region that links the N-terminal DNA-binding domain and the C-terminal transcriptional domain. Phosphatases that dephosphorylate the linker region are therefore likely to play an integral part in the regulation of Smad activity. We reported previously that small C-terminal domain phosphatases 1, 2, and 3 (SCP1-3) dephosphorylate Smad1 C-terminal tail, thereby attenuating BMP signaling. Here we provide evidence that SCP1-3 also dephosphorylate the linker regions of Smad1 and Smad2/3 in vitro, in mammalian cells and in Xenopus embryos. Overexpression of SCP 1, 2, or 3 decreased linker phosphorylation of Smads 1, 2 and 3. Moreover, RNA interference-mediated knockdown of SCP1/2 increased the BMP-dependent phosphorylation of the Smad1 linker region as well as the C terminus. In contrast, SCP1/2 knockdown increased the TGFbeta-dependent linker phosphorylation of Smad2/3 but not the C-terminal phosphorylation. Consequently, SCP1/2 knockdown inhibited TGFbeta transcriptional responses, but it enhanced BMP transcriptional responses. Thus, by dephosphorylating Smad2/3 at the linker (inhibitory) but not the C-terminal (activating) site, the SCPs enhance TGFbeta signaling, and by dephosphorylating Smad1 at both sites, the SCPs reset Smad1 to the basal unphosphorylated state. PMID- 17085435 TI - Ligand-induced structural changes of the CD44 hyaluronan-binding domain revealed by NMR. AB - CD44, a major cell surface receptor for hyaluronan (HA), contains a functional domain responsible for HA binding at its N terminus (residues 21-178). Accumulating evidence indicates that proteolytic cleavage of CD44 in its extracellular region (residues 21-268) leads to enhanced tumor cell migration and invasion. Hence, understanding the mechanisms underlying the CD44 proteolytic cleavage is important for understanding the mechanism of CD44-mediated tumor progression. Here we present the NMR structure of the HA-binding domain of CD44 in its HA-bound state. The structure is composed of the Link module (residues 32 124) and an extended lobe (residues 21-31 and 125-152). Interestingly, a comparison of its unbound and HA-bound structures revealed that rearrangement of the beta-strands in the extended lobe (residues 143-148) and disorder of the structure in the following C-terminal region (residues 153-169) occurred upon HA binding, which is consistent with the results of trypsin proteolysis studies of the CD44 HA-binding domain. The order-to-disorder transition of the C-terminal region by HA binding may be involved in the CD44-mediated cell migration. PMID- 17085436 TI - IQGAP1 stimulates actin assembly through the N-WASP-Arp2/3 pathway. AB - IQGAP1 is a conserved modular protein overexpressed in cancer and involved in organizing actin and microtubules in motile processes such as adhesion, migration, and cytokinesis. A variety of proteins have been shown to interact with IQGAP1, including the small G proteins Rac1 and Cdc42, actin, calmodulin, beta-catenin, the microtubule plus end-binding proteins CLIP170 (cytoplasmic linker protein) and adenomatous polyposis coli. However, the molecular mechanism by which IQGAP1 controls actin dynamics in cell motility is not understood. Quantitative co-localization analysis and down-regulation of IQGAP1 revealed that IQGAP1 controls the co-localization of N-WASP with the Arp2/3 complex in lamellipodia. Co-immunoprecipitation supports an in vivo link between IQGAP1 and N-WASP. Pull-down experiments and kinetic assays of branched actin polymerization with N-WASP and Arp2/3 complex demonstrated that the C-terminal half of IQGAP1 activates N-WASP by interacting with its BR-CRIB domain in a Cdc42-like manner, whereas the N-terminal half of IQGAP1 antagonizes this activation by association with a C-terminal region of IQGAP1. We propose that signal-induced relief of the autoinhibited fold of IQGAP1 allows activation of N-WASP to stimulate Arp2/3 dependent actin assembly. PMID- 17085437 TI - Integrin regulation by vascular endothelial growth factor in human brain microvascular endothelial cells: role of alpha6beta1 integrin in angiogenesis. AB - The precise role of vascular endothelial growth factor (VEGF) in regulating integrins in brain microvascular endothelial cells is unknown. Here, we analyzed VEGF effects on integrin expression and activation in human brain microvascular endothelial cells (HBMECs). Using human cDNA arrays and ribonuclease (RNase) protection assays, we observed that VEGF up-regulated the mRNA expression of alpha(6) integrin in HBMECs. VEGF significantly increased alpha(6)beta(1) integrin expression, but not alpha(6)beta(4) integrin expression in these cells. Specific down-regulation of alpha(6) integrin expression by small interfering RNA (siRNA) oligonucleotides inhibited both the capillary morphogenesis of HBMECs and their adhesion and migration. Additionally, VEGF treatment resulted in activation of alpha(6)beta(1) integrins in HBMECs. Functional blocking of alpha(6) integrin with its specific antibody inhibited the VEGF-induced adhesion and migration as well as in vivo angiogenesis, and markedly suppressed tumor angiogenesis and breast carcinoma growth in vivo. Thus, VEGF can modulate angiogenesis via increased expression and activation of alpha(6)beta(1) integrins, which may promote VEGF-driven tumor angiogenesis in vivo. PMID- 17085438 TI - Differential expression of the B'beta regulatory subunit of protein phosphatase 2A modulates tyrosine hydroxylase phosphorylation and catecholamine synthesis. AB - Tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine synthesis, is stimulated by N-terminal phosphorylation by several kinases and inhibited by protein serine/threonine phosphatase 2A (PP2A). PP2A is a family of heterotrimeric holoenzymes containing one of more than a dozen different regulatory subunits. In comparison with rat forebrain extracts, adrenal gland extracts exhibited TH hyperphosphorylation at Ser(19), Ser(31), and Ser(40), as well as reduced phosphatase activity selectively toward phosphorylated TH. Because the B'beta regulatory subunit of PP2A is expressed in brain but not in adrenal glands, we tested the hypothesis that PP2A/B'beta is a specific TH phosphatase. In catecholamine-secreting PC12 cells, inducible expression of B'beta decreased both N-terminal Ser phosphorylation and in situ TH activity, whereas inducible silencing of endogenous B'beta had the opposite effect. Furthermore, PP2A/B'beta directly dephosphorylated TH in vitro. As to specificity, other PP2A regulatory subunits had negligible effects on TH activity and phosphorylation in situ and in vitro. Whereas B'beta was highly expressed in dopaminergic cell bodies in the substantia nigra, the PP2A regulatory subunit was excluded from TH-positive terminal fields in the striatum and failed to colocalize with presynaptic markers in general. Consistent with a model in which B'beta enrichment in neuronal cell bodies helps confine catecholamine synthesis to axon terminals, TH phosphorylation was higher in processes than in somata of dopaminergic neurons. In summary, we show that B'beta recruits PP2A to modulate TH activity in a tissue- and cell compartment specific fashion. PMID- 17085439 TI - Structural basis for the platelet-collagen interaction: the smallest motif within collagen that recognizes and activates platelet Glycoprotein VI contains two glycine-proline-hydroxyproline triplets. AB - Collagen-related peptide is a selective agonist for the platelet collagen receptor Glycoprotein VI. The triple helical peptide contains ten GPO triplets/strand (single letter amino acid nomenclature, where O is hydroxyproline) and so over-represents GPO compared with native collagen sequence. To investigate the ability of Glycoprotein VI to recognize GPO triplets in a setting more representative of the collagens, we synthesized a set of triple helical peptides containing fewer GPO triplets, varying their number and spacing within an inert (GPP)n backbone. The adhesion of recombinant human Glycoprotein VI ectodo-main, like that of human platelets, to these peptides increased with their GPO content, and platelet adhesion was abolished by the specific anti Glycoprotein VI-blocking antibody, 10B12. Platelet aggregation and protein tyrosine phosphorylation were induced only by cross-linked peptides and only those that contained two or more GPO triplets. Such peptides were less potent than cross-linked collagen-related peptide. Our data suggest that both the sequences GPOGPO and GPO.........GPO represent functional Glycoprotein VI recognition motifs within collagen. Furthermore, we propose that the (GPO)4 motif can support simultaneous binding of two glycoprotein VI molecules, in either a parallel or anti-parallel stacking arrangement, which could play an important role in activation of signaling. PMID- 17085440 TI - Deregulation of AP-1 proteins in collagen gel-induced epithelial cell apoptosis mediated by low substratum rigidity. AB - In this study, we established that collagen gel, but not collagen gel coating, induced apoptosis exclusively in epithelial cell lines, which indicated that low substratum rigidity might trigger cell apoptosis. To confirm this, we used collagen gels with different rigidities due to cross-linking or physical disruption of collagen fibrils caused by sonication. We found that collagen gel induced apoptosis was inversely correlated with substratum rigidity. Low substratum rigidity collagen gel-induced apoptosis was neither prevented by Bcl-2 overexpression nor preceded by mitochondrial release of cytochrome c. This suggested that the mitochondrial pathway was not involved in low substratum rigidity-induced apoptosis. Low substratum rigidity activated c-Jun N-terminal kinase (JNK) within 4 h, but it also rapidly down-regulated c-Jun within 1 h and triggered persistent aberrant expression of c-Fos for at least 24 h. Either reduced c-Jun expression or c-Fos overexpression induced apoptosis in several epithelial cells. Inhibiting low substratum rigidity-induced JNK activation prevented aberrant c-Fos expression but only partially blocked low substratum rigidity-induced apoptosis. Taking these results together, we conclude that low substratum rigidity collagen gel induced apoptosis in epithelial cells and that deregulated AP-1 proteins mediated that apoptosis, at least in part. PMID- 17085441 TI - Substrate recognition by RNA 5-methyluridine methyltransferases and pseudouridine synthases: a structural perspective. PMID- 17085442 TI - Feeding and insulin increase leptin translation. Importance of the leptin mRNA untranslated regions. AB - The post-transcriptional mechanisms by which feeding and insulin increase leptin production are poorly understood. Starvation of 6-7-week-old rats for 14 h decreased leptin mRNA level by only 22% but decreased plasma levels, adipose tissue leptin content, and release by over 75%. The decreased leptin with starvation was explained by >85% decrease in relative rates of leptin biosynthesis measured by metabolic labeling and immunoprecipitation. In vitro insulin treatment of adipose tissue from fed or starved rats for 2 h increased relative rates of leptin biosynthesis by 2-3-fold, and the effect was blocked by inhibition of phosphatidylinositol 3-kinase or mammalian target of rapamycin. Consistent with the hypothesis that feeding/insulin increases leptin translation, more leptin mRNA was associated with polysomes in adipose tissue of fed than starved rats, and in vitro incubation of adipose tissue of starved rats with insulin shifted leptin mRNA into polysomes. To assess the mechanisms regulating leptin translation, chimeric human leptin untranslated region (UTR) reporter constructs were transiently transfected into differentiated 3T3-L1 adipocytes. The 5'-UTR of leptin mRNA increased luciferase reporter activity 2-3-fold, whereas the full-length 3'-UTR (nucleotides 1-2804) was inhibitory (-65%). Sequences between nucleotides 462 and 1130 of the leptin 3'-UTR conferred most of the inhibitory effect. Insulin stimulated the expression of constructs that included both the full-length 5'-UTR and the inhibitory 3'-UTR, and the effect was blocked by inhibition of phosphatidylinositol 3-kinase or mammalian target of rapamycin. Our data suggest that insulin derepresses leptin translation by a mechanism that requires both the 5'-UTR and the 3'-UTR and may contribute to the increase in leptin production with feeding. PMID- 17085443 TI - VMD2 promoter requires two proximal E-box sites for its activity in vivo and is regulated by the MITF-TFE family. AB - The retinal pigment epithelium (RPE) is crucial for the function and survival of retinal photoreceptors. VMD2 encodes bestrophin, an oligomeric chloride channel that is preferentially expressed in the RPE and, when mutated, causes Best macular dystrophy. Previously, we defined the VMD2 upstream region from -253 to +38 bp as being sufficient to direct RPE-specific expression in the eye, and we suggested microphthalmia-associated transcription factor (MITF) as a possible positive regulator. Here we show that in transgenic mice the -154 to +38 bp region is sufficient for RPE expression, and mutation of two E-boxes, 1 and 2, within this region leads to loss of promoter activity. A yeast one-hybrid screen using bait containing E-box 1 identified clones encoding MITF, TFE3, and TFEB, and chromatin immunoprecipitation with antibodies against these proteins enriched the VMD2 proximal promoter. Analysis using in vivo electroporation with constructs containing mutation of each E-box indicated that expression in native RPE requires both E-boxes, yet in vitro DNA binding studies suggested that MITF binds well to E-box 1 but only minimally to E-box 2. MITF knockdown by small interfering RNA (siRNA) in cell culture revealed a strong correlation between MITF and VMD2 mRNA levels. Sequential transfection of a luciferase construct with expression vectors following MITF siRNA revealed that TFE3 and TFEB can also transactivate the VMD2 promoter. Taken together, we suggest that VMD2 is regulated by the MITF-TFE family through two E-boxes, with E-box 1 required for a direct interaction of MITF-TFE factors and E-box 2 for binding of the as yet unidentified factor(s). PMID- 17085444 TI - Inner nuclear membrane proteins Asi1, Asi2, and Asi3 function in concert to maintain the latent properties of transcription factors Stp1 and Stp2. AB - In yeast the homologous transcription factors Stp1 and Stp2 are synthesized as latent cytoplasmic precursors with N-terminal regulatory domains. In response to extracellular amino acids the regulatory domains are endoproteolytically excised by the plasma membrane-localized SPS sensor. The processed forms of Stp1 and Stp2 efficiently enter the nucleus and induce expression of amino acid permease genes. We recently reported that the inner nuclear membrane protein Asi1 is required to prevent unprocessed forms of Stp1 and Stp2, which ectopically enter the nucleus, from binding SPS sensor-regulated promoters. Here we show that Asi3, an Asi1 homolog, and Asi2 are integral proteins of the inner nuclear membrane that function in concert with Asi1. In cells lacking any of the three Asi proteins, unprocessed full-length forms of Stp1 and Stp2 constitutively induce SPS sensor regulated genes. Our results demonstrate that the Asi proteins ensure the fidelity of SPS sensor signaling by maintaining the dormant, or repressed state, of gene expression in the absence of inducing signals. This study documents additional components of a novel mechanism controlling transcription in eukaryotic cells. PMID- 17085445 TI - Characterization of the Interleukin (IL)-6 Inhibitor IL-6-RFP: fused receptor domains act as high affinity cytokine-binding proteins. AB - Although fusion proteins of the extracellular parts of receptor subunits termed cytokine traps turned out to be promising cytokine inhibitors for anti-cytokine therapies, their mode of action has not been analyzed. We developed a fusion protein consisting of the ligand binding domains of the IL-6 receptor subunits IL 6Ralpha and gp130 that acts as a highly potent IL-6 inhibitor. Gp130 is a shared cytokine receptor also used by the IL-6-related cytokines oncostatin M and leukemia inhibitory factor. In this study, we have shown that the IL-6 receptor fusion protein (IL-6-RFP) is a specific IL-6 inhibitor that does not block oncostatin M or leukemia inhibitory factor. We characterized the complex of IL-6 RFP and fluorescently labeled IL-6 (YFPIL-6) by blue native PAGE and gel filtration. A 2-fold molar excess of IL-6-RFP over IL-6 was sufficient to entirely bind IL-6 in a complex with IL-6-RFP. As shown by treatment with urea and binding competition experiments, the complex of IL-6 and IL-6-RFP is more stable than the complex of IL-6, soluble IL-6Ralpha, and soluble gp130. By live cell imaging, we have demonstrated that YFP-IL-6 bound to the surface of cells expressing gp130-CFP is removed from the plasma membrane upon the addition of IL 6-RFP. The apparent molecular mass of the IL-6.IL-6-RFP complex determined by blue native PAGE and gel filtration suggests that IL-6 is trapped in a structure analogous to the native hexameric IL-6 receptor complex. Thus, fusion of the ligand binding domains of heteromeric receptors leads to highly specific cytokine inhibitors with superior activity compared with the separate soluble receptors. PMID- 17085446 TI - Calcium-mediated transient phosphorylation of tau and amyloid precursor protein followed by intraneuronal amyloid-beta accumulation. AB - Intraneuronal accumulation of hyperphosphorylated protein tau in paired helical filaments together with amyloid-beta peptide (Abeta) deposits confirm the clinical diagnosis of Alzheimer disease. A common cellular mechanism leading to the production of these potent toxins remains elusive. Here we show that, in cultured neurons, membrane depolarization induced a calcium-mediated transient phosphorylation of both microtubule-associated protein tau and amyloid precursor protein (APP), followed by a dephosphorylation of these proteins. Phosphorylation was mediated by glycogen synthase kinase 3 and cyclin-dependent kinase 5 protein kinases, while calcineurin was responsible for dephosphorylation. Following the transient phosphorylation of APP, intraneuronal Abeta accumulated and induced neurotoxicity. Phosphorylation of APP on Thr-668 was indispensable for intraneuronal accumulation of Abeta. Our data demonstrate that an increase in cytosolic calcium concentration induces modifications of neuronal metabolism of APP and tau, similar to those found in Alzheimer disease. PMID- 17085447 TI - Structural basis for p53 binding-induced DNA bending. AB - Specific p53 binding-induced DNA bending has important biological implications such as transcription activation. However, the detailed structures of the bent DNA and the p53-DNA complex are still unavailable, hampering our understanding of the mechanism for p53-induced DNA bending and its consequent biological significance. To gain insight into the p53 binding-induced DNA bending, we performed molecular dynamics simulations on DNA segments with the consensus sequence for p53-specific binding, half site DNA-p53 complexes, and full site DNA p53 complexes. We show that each DNA-bound p53 core domain caused a local DNA conformational change within the quarter site; upon the binding of the p53 dimer, there was an apparent DNA bending at the center of the half site; when bound with two p53 dimers, the full site DNAs with two different sequences bent 20 and 35 degrees, respectively. These results are in agreement with experimental observations. Our simulations demonstrate that the two p53 dimers favored a staggered conformation in which they make favorable interactions at the interface. This dimer-dimer interface organization necessitated conformational changes in the DNA, leading to the bending at the center of the full site, which in turn is dependent on the DNA sequence. Overall, our results provide the detailed atomic model for the DNA-p53 tetramer complex and delineate the roles of DNA-p53, p53 dimer-dimer interactions, and DNA sequence in specific p53 binding induced DNA conformational changes. PMID- 17085448 TI - Residues His-15 and Arg-17 of HPr participate differently in catabolite signal processing via CcpA. AB - The carbon catabolite control protein A (CcpA) senses the physiological state of the cell by binding several effectors and responds with differential regulation of many genes in Bacilli. HPr-Ser46-P or Crh-Ser46-P interact with CcpA and stimulate binding to catabolite responsive elements. In addition, the glycolytic intermediates fructose 1,6-bisphosphate (FBP) and glucose 6-phosphate (Glc-6-P) stimulate HPr-Ser46-P but not Crh-Ser46-P binding to CcpA. The mechanisms by which coeffector binding to CcpA is linked to differential gene expression are unclear. To address this question we mutated residues participating in the interaction between HPr-Ser46-P or Crh-Ser46-P and CcpA and analyzed their effects on CcpA binding and stimulation of cre binding by surface plasmon resonance. The HPrH15A and CcpAD297A mutations do not affect complex formation but abolish FBP and Glc-6-P stimulation. Likewise, the CrhQ15H mutant becomes sensitive to these glycolytic intermediates. Hence, the contact of HPrHis-15 to Asp-297 in CcpA is a determinant for HPr specific FBP and Glc-6-P stimulation. The HPrR17A and -K mutants are both strongly impaired in stimulation of CcpA binding to cre, but only HPrR17A is defect in binding to CcpA indicating that these residues affect allostery of CcpA. Mutations of the residues of CcpA, which contact Arg-17 of HPr, exhibit differential effects on regulation of catabolic genes. Taken together, His-15 of HPr processes sensing information, while Arg-17 is involved in determining the genetic output. PMID- 17085449 TI - The role of the ubiquitin ligase E6-AP in human papillomavirus E6-mediated degradation of PDZ domain-containing proteins. AB - The E6 oncoprotein of human papillomaviruses associated with cervical cancer targets the tumor suppressor p53 and several other cellular proteins including the human homologs of Dlg and Scribble for degradation via the ubiquitin proteasome system. Similar to p53 degradation, E6-induced degradation of Scribble is mediated by the ubiquitin ligase E6-AP. In contrast, degradation of Dlg in vitro and within cells has been reported to be independent of E6-AP, suggesting that the E6 oncoprotein has the ability to interact with ubiquitin ligases other than E6-AP. Furthermore, the ability of the E6 oncoprotein to interact with these yet unidentified ubiquitin ligases may be shared by the E6 protein of so-called low risk human papillomaviruses that are not associated with cervical cancer. In this study, we used the RNA interference technology and mouse embryo fibroblasts derived from E6-AP-deficient mice to obtain information about the identity of the ubiquitin ligase(s) involved in E6-mediated degradation of Dlg. We report that, within cells, E6-mediated degradation of Dlg depends on the presence of functional E6-AP and provide evidence that the E6 protein of low risk human papillomaviruses functionally interacts with E6-AP. Based on these data, we propose that, in general, the proteolytic properties of human papillomavirus E6 proteins are mediated by interaction with E6-AP. PMID- 17085450 TI - The anti-rheumatic gold salt aurothiomalate suppresses interleukin-1beta-induced hyaluronan accumulation by blocking HAS1 transcription and by acting as a COX-2 transcriptional repressor. AB - Gold compounds are among the oldest disease-modifying drugs and are still widely used today for treating rheumatoid arthritis. Despite decades of use, little is known about the mode of action of this class of drugs. Here we have demonstrated that aurothiomalate (AuTM) suppresses hyaluronan accumulation by blocking interleukin (IL)-1beta-induced hyaluronan synthase-1 transcription. We have further demonstrated that, in fibroblast-like synoviocytes (FLSs), AuTM acts as a specific COX-2 transcriptional repressor in that IL-1beta-induced COX-2 transcription is blocked, whereas COX-1 transcription and translation is unaffected. As a consequence, PGE2 levels released by FLS are dose-dependently reduced in cells exposed to AuTM. Of similar importance is the demonstration that AuTM does block NFkappaB-DNA interaction. In addition, two other transcription factors implicated in inflammatory events, namely AP-1 and STAT3, are blocked as well. The effect on NFkappaB likely explains the inhibition of COX-2 as well as that of HAS1, as both are genes that depend on the activation of NFkappaB. Interestingly, AuTM does not interfere with IL-1beta-induced IkappaB alpha degradation, in most cases a prerequisite for subsequent NFkappaB activation. Furthermore, evidence is presented that, in FLS, AuTM blocks NFkappaB-DNA interaction neither by binding to NFkappaB binding sites nor by interacting with activated NFkappaB proteins. Taken together, AuTM treatment of FLS blocks two of the most important proinflammatory events that are associated with rheumatoid arthritis. AuTM blocks the release of PGE2 and prevents the activation of NFkappaB, therefore blocking IL-1beta-induced hyaluronan accumulation and likely a series of other pro-inflammatory NFkappaB-dependent genes. PMID- 17085451 TI - Structural basis for the adherence of Plasmodium falciparum-infected erythrocytes to chondroitin 4-sulfate and design of novel photoactivable reagents for the identification of parasite adhesive proteins. AB - A dodecasaccharide motif of the low-sulfated chondroitin 4-sulfate (C4S) mediate the binding of Plasmodium falciparum-infected red blood cells (IRBCs) in human placenta. Here we studied the detailed C4S structural requirements by assessing the ability of chemically modified C4S to inhibit IRBC binding to the placental chondroitin sulfate proteoglycan. Replacement of the N-acetyl groups with bulky N acyl or N-benzoyl substituents had no effect on the inhibitory activity of C4S, whereas reduction of the carboxyl groups abrogated the activity. Dermatan sulfates showed approximately 50% inhibitory activity when compared with C4Ss with similar sulfate contents. These data demonstrate that the C4S carboxyl groups and their equatorial orientation but not the N-acetyl groups are critical for IRBC binding. Conjugation of bulky substituents to the reducing end N acetylgalactosamine residues of C4S dodecasaccharide had no effect on its inhibitory activity. Based on these results, we prepared photoaffinity reagents for the identification of the parasite proteins involved in C4S binding. Cross linking of the IRBCs with a radioiodinated photoactivable C4S dodecasaccharide labeled a approximately 22-kDa novel parasite protein, suggesting strongly for the first time that a low molecular weight IRBC surface protein rather than a 200 400-kDa PfEMP1 is involved in C4S binding. Conjugation of biotin to the C4S dodecasaccharide photoaffinity probe afforded a strategy for the isolation of the labeled protein by avidin affinity precipitation, facilitating efforts to identify the C4S-adherent IRBC protein(s). Our results also have broader implications for designing oligosaccharide-based photoaffinity probes for the identification of proteins involved in glycosaminoglycan-dependent attachment of microbes to hosts. PMID- 17085452 TI - Beta-catenin signaling pathway is crucial for bone morphogenetic protein 2 to induce new bone formation. AB - Endochondral ossification is recapitulated during bone morphogenetic protein (BMP)-induced ectopic bone formation. Although BMP and beta-catenin have been investigated in bone development and in mesenchymal cells, how they interact in this process is not clear. We implanted recombinant BMP-2 into the muscle of mice to investigate the effect of beta-catenin signaling on BMP-induced in vivo endochondral bone formation. BMP-2 induced expression of several Wnt ligands and their receptors and also activated beta-catenin-mediated T cell factor-dependent transcriptional activity. An adenovirus expressing Dickkopf-1 (Dkk-1, an inhibitor of canonical Wnt pathway) inhibited beta-catenin signaling and endochondral bone formation. Interestingly, Dkk-1 inhibited both chondrogenesis and osteogenesis. Likewise, mice expressing conditional beta-catenin null alleles also displayed an inhibition of BMP-induced chondrogenesis and osteogenesis. This is in contrast to studies of embryonic skeletogenesis, which demonstrate that beta-catenin is required for osteogenesis but is dispensable for chondrogenesis. These findings suggest that embryonic development pathways are not always recapitulated during post-natal regenerative processes, and the biochemical pathways utilized to regulate cell differentiation may be different. During in vivo ectopic bone formation, BMP-2 induces beta-catenin-mediated signaling through Wnt ligands, and beta-catenin is required for both chondrogenesis and osteogenesis. PMID- 17085453 TI - AlphaCP1 mediates stabilization of hTERT mRNA by autocrine human growth hormone. AB - We herein demonstrate that autocrine human growth hormone production in human mammary carcinoma cells results in increased telomerase activity as a result of specific up-regulation of telomerase catalytic subunit (human telomerase reverse transcriptase (hTERT)) mRNA and protein. This increase in hTERT gene expression is not due to increased transcriptional activation of the hTERT promoter but is the result of increased stability of hTERT mRNA exerted by CU-rich cis-regulatory sequences present in the 3'-untranslated region of TERT mRNA. Autocrine human growth hormone up-regulates two poly(C)-binding proteins, alphaCP1 and alphaCP2, which bind to these cis-regulatory elements and stabilize hTERT mRNA. We have therefore demonstrated that post-transcriptional modulation of the level of hTERT mRNA is one mechanism for regulation of cellular telomerase activity. PMID- 17085454 TI - Commentary: the health crisis in the USSR: looking behind the facade. PMID- 17085455 TI - Effect of education on overall and cause-specific mortality in injecting drug users, according to HIV and introduction of HAART. AB - BACKGROUND: We assessed the impact of education on long-term overall and cause specific mortality among 6575 injecting drug users (IDUs) according to HIV status and introduction of highly active antiretroviral therapy (HAART). METHODS: Community-based cohort study of IDUs recruited in three AIDS prevention centres (1987-1996). Causes of death were ascertained in clinical centres and Mortality Registry and classified as AIDS, drug use related, injuries, or liver diseases. Poisson regression models including education and calendar period interaction and adjusted by sex, age, and HIV were used. RESULTS: In 73 901 person-years of follow-up, there were 1493 deaths (20.2/1000 person-years): 761 related to AIDS, 234 to drug use, 179 to injuries, and 93 to liver diseases. IDUs with university studies had a lower risk of death (RR 0.52; 95% CI 0.36-0.77) than those without studies: this difference was higher after (RR 0.45; 95% CI 0.25-0.80) than before 1997 (RR 0.68; 95% CI 0.41-1.13). Compared to before 1997, while decreases in the risk of AIDS mortality were seen during 1997-2004 for both lower (RR 0.49; 95% CI 0.41-0.58) and higher (RR 0.33; 95% CI 0.23-0.48) educated, only those higher educated experienced a reduction in drug-use mortality (RR 0.54; 95% CI 0.28 1.05) and death from injuries (RR 0.52; 95% CI 0.23-1.21). CONCLUSIONS: Independently of HIV status, lower education predicts a higher risk of death in IDUs and its impact is stronger after 1997. Education has a protective effect on most causes of death and it cannot be entirely attributable to the access or use of HAART. PMID- 17085456 TI - Does biofuel smoke contribute to anaemia and stunting in early childhood? AB - BACKGROUND: Reliance on biomass fuels for cooking and heating exposes many women and young children in developing countries to high levels of air pollution indoors. Exposure to biomass smoke has been linked to reduced birth weight, acute respiratory infections, and childhood mortality. This study examines the association between household use of biofuels (wood, dung, and crop residues) for cooking and heating and prevalence of anaemia and stunting in children. METHODS: Data are from a 1998-99 national family health survey in India, which measured height, weight, and blood haemoglobin of 29 768 children aged 0-35 months in 92 486 households. Multinomial logistic regression is used to estimate the effects of biofuel use on prevalence of anaemia and stunting, controlling for exposure to tobacco smoke, recent episodes of illness, maternal education and nutrition, and other potentially confounding factors. RESULTS: Analysis shows that prevalence of moderate-to-severe anaemia was significantly higher among children in households using biofuels than among children in households using cleaner fuels (RRR = 1.58; 95% CI: 1.28, 1.94), independent of other factors. Prevalence of severe stunting was also significantly higher among children in biofuel-using households (RRR = 1.84; 95% CI: 1.44, 2.36). Thirty-one per cent of moderate-to-severe anaemia and 37% of severe stunting among children aged 6-35 months in India may be attributable to exposure to biofuel smoke. Effects on mild anaemia and moderate stunting were smaller, but positive and statistically significant. Effects of exposure to tobacco smoke on anaemia and stunting were small and not significant. CONCLUSIONS: The study provides a first evidence of the strong association between biofuel use and risks of anaemia and stunting in children, suggesting that exposure to biofuel smoke may contribute to chronic nutritional deficiencies in young children. PMID- 17085457 TI - Socioeconomic differentials in cause-specific mortality among South Korean adolescents. AB - BACKGROUND: There is inconsistent evidence regarding the presence of a socioeconomic differential in adolescent all-cause and cause-specific mortality. This study examines possible socioeconomic mortality differentials in Korean adolescents. Method A total of 330 321 boys and 311 830 girls aged 10-19, who are health insurance beneficiaries for civil servants and private school teachers of Korean Health Insurance Cooperation, were followed for 9 years (1995-2003). Parental income information was linked to national death certificate data. RESULTS: For boys, all-cause mortality showed a graded inverse relationship with income level in both 10-14 year olds (RR = 1.64, 95% CI: 1.40-1.91) and 15-19 year olds (RR = 1.68, 95% CI: 1.40-1.91). The major contributor was mortality differentials from external causes, with differentials of transport accident death the most important. Mortality from circulatory disease was higher in the lowest income groups in 15-19 year olds (RR = 2.21, 95% CI: 1.09-4.50). A significant socioeconomic gradient of non-external cause mortality was found in 15-19 year olds. For girls, socioeconomic differentials were less evident than boys. The all-cause mortality gradient for girls was smaller than for boys and only significant between the lowest and the highest tertile in both 10-14 year olds and 15-19 year olds (RR = 1.33, 95% CI: 1.02-1.72, RR = 1.38, 95% CI: 1.11 1.72, respectively). There were significant socioeconomic mortality differentials in all external causes and transport accidents and a marginally significant difference in suicide mortality for 10-19 year olds. Mortality from non-external causes showed no social gradient in girls. CONCLUSIONS: Socioeconomic differentials in all-cause mortality were observed in adolescents, even in early youth. This pattern might also apply to mortality from non-external causes, especially cardiovascular disease in 15-19 year old males. PMID- 17085458 TI - Endurance training and detraining in mitochondrial myopathies due to single large scale mtDNA deletions. AB - At present there are limited therapeutic interventions for patients with mitochondrial myopathies. Exercise training has been suggested as an approach to improve physical capacity and quality of life but it is uncertain whether it offers a safe and effective treatment for patients with heteroplasmic mitochondrial DNA (mtDNA) mutations. The objectives of this study were to assess the effects of exercise training and detraining in eight patients with single, large-scale mtDNA deletions to determine: (i) the efficacy and safety of endurance training (14 weeks) in this patient population; (ii) to determine the effect of more prolonged (total of 28 weeks) exercise training upon muscle and cardiovascular function and (iii) to evaluate the effect of discontinued training (14 weeks) upon muscle and cardiovascular function. Our results show that: (i) 14 weeks of exercise training significantly improved tolerance of submaximal exercise and peak capacity for work, oxygen utilization and skeletal muscle oxygen extraction with no change in the level of deleted mtDNA; (ii) continued training for an additional 14 weeks maintained these beneficial adaptations; (iii) the cessation of training (detraining) resulted in loss of physiological adaptation to baseline capacity with no overall change in mutation load. Patients' self assessment of quality of life as measured by the SF-36 questionnaire improved with training and declined with detraining. Whilst our findings of beneficial effects of training on physiological outcome and quality of life without increases in the percentage of deleted mtDNA are encouraging, we did not observe changes in mtDNA copy number. Therefore there remains a need for longer term studies to confirm that endurance exercise is a safe and effective treatment for patients with mitochondrial myopathies. The effects of detraining clearly implicate physical inactivity as an important mechanism in reducing exercise capacity and quality of life in patients with mitochondrial myopathy. PMID- 17085459 TI - Effect of isoniazid prophylaxis on mortality and incidence of tuberculosis in children with HIV: randomised controlled trial. AB - OBJECTIVES: To investigate the impact of isoniazid prophylaxis on mortality and incidence of tuberculosis in children with HIV. DESIGN: Two centre prospective double blind placebo controlled trial. PARTICIPANTS: Children aged > or =8 weeks with HIV. INTERVENTIONS: Isoniazid or placebo given with co-trimoxazole either daily or three times a week. SETTING: Two tertiary healthcare centres in South Africa. MAIN OUTCOME MEASURES: Mortality, incidence of tuberculosis, and adverse events. RESULTS: Data on 263 children (median age 24.7 months) were available when the data safety monitoring board recommended discontinuing the placebo arm; 132 (50%) were taking isoniazid. Median follow-up was 5.7 (interquartile range 2.0-9.7) months. Mortality was lower in the isoniazid group than in the placebo group (11 (8%) v 21 (16%), hazard ratio 0.46, 95% confidence interval 0.22 to 0.95, P=0.015) by intention to treat analysis. The benefit applied across Centers for Disease Control clinical categories and in all ages. The reduction in mortality was similar in children on three times a week or daily isoniazid. The incidence of tuberculosis was lower in the isoniazid group (5 cases, 3.8%) than in the placebo group (13 cases, 9.9%) (hazard ratio 0.28, 0.10 to 0.78, P=0.005). All cases of tuberculosis confirmed by culture were in children in the placebo group. CONCLUSIONS: Prophylaxis with isoniazid has an early survival benefit and reduces incidence of tuberculosis in children with HIV. Prophylaxis may offer an effective public health intervention to reduce mortality in such children in settings with a high prevalence of tuberculosis. TRIAL REGISTRATION: Clinical Trials NCT00330304. PMID- 17085460 TI - Parent and family factors associated with child adjustment to pediatric cancer. AB - OBJECTIVE: To identify factors that influence the association between parent and child distress among families of children with cancer and comparison peers. METHODS: Parent and child distress, social support, and family environment were assessed among families of 95 children with cancer (94 mothers, 67 fathers) and 98 comparison peers (97 mothers, 77 fathers). RESULTS: Significant associations were found between parent and child distress. For models examining the impact of fathers' distress on children, several moderators were identified (i.e., family environment, child age and gender, a cancer diagnosis, and treatment severity). Family environment also partially mediated father and child distress. CONCLUSIONS: Children whose parents were distressed were more likely to be distressed themselves. Subgroups of children were particularly vulnerable, indicating a need to identify further mechanisms of risk and resilience and to develop family-based interventions. Support was found for including fathers as independent sources of information in pediatric psychology research and clinical practice. PMID- 17085461 TI - Candesartan reduces cardiovascular events in patients on chronic haemodialysis. PMID- 17085462 TI - The nurse, mother of two and four transplants--Nancy Spaeth tells her story. PMID- 17085463 TI - Renoprotective effect of COMP-angiopoietin-1 in db/db mice with type 2 diabetes. AB - BACKGROUND: Inflammatory processes have been recently seen as underlying the pathogenesis of diabetic nephropathy. Angiopoietin-1 (Ang1) plays essential roles in regulating vascular growth, development, maturation, permeability and inflammation. We have developed a soluble, stable and potent Ang1 variant, cartilage oligomeric matrix protein (COMP)-Ang1. METHODS: In this study, db/db mice were treated with recombinant adenovirus expressing either COMP-Ang1 or LacZ. Histology, inflammatory, metabolic, and fibrotic parameters and signalling pathway were evaluated. RESULTS: COMP-Ang1 reduced albuminuria and decreased mesangial expansion, thickening of the glomerular basement membrane and podocyte foot process broadening and effacement. COMP-Ang1 suppressed both renal expression of intercellular adhesion molecule-1 and monocyte chemoattractant protein-1 and monocyte/macrophage infiltration in diabetic db/db mice. COMP-Ang1 also reduced renal tissue levels of transforming growth factor-beta1 (TGF-beta1), alpha-smooth muscle actin, fibronectin, as well as Smad 2/3 expression, but increased Smad 7 expression. In human umbilical vein endothelial cells (HUVECs) grown in high glucose concentrations of glucose, recombinant COMP-Ang1 protein decreased nuclear factor-kappaB (NF-kappaB) expression. COMP-Ang1-mediated inhibition of increased NF-kappaB-DNA binding in nuclear extracts from HUVECs grown in high glucose was significantly blocked by soluble Tie2 receptor-Fc. In addition, COMP-Ang1 significantly decreased fasting blood glucose level, epididymal fat weight to body weight ratio, and epididymal adipocyte size in diabetic db/db mice. After intraperitoneal glucose challenge, COMP-Ang1 significantly lowered plasma glucose levels. However, there was no difference in serum insulin levels. CONCLUSION: We conclude that COMP-Ang1 delayed the fibrotic changes in the kidney of diabetic db/db mice through its anti-inflammatory or metabolic effects. PMID- 17085464 TI - NADPH oxidase subunits (NOX-1, p22phox, Rac-1) and tacrolimus-induced nephrotoxicity in a rat renal transplant model. AB - BACKGROUND: TGF-beta and oxidative stress are known mediators of renal injury. However, the precise mechanisms by which TGF-beta and oxidative stress may be involved in the development of nephrotoxicity are not known. We examined whether anti-TGF-beta antibody limits nephrotoxicity produced by tacrolimus (TAC) and whether this altered genes that regulate oxidative stress. METHODS: Renal transplants were performed in Wistar-Furth and Lewis rat strains. Groups included: isograft controls; untreated allografts; allografts treated with 0.25 mg/kg TAC till 90 days with or without 1.0 mg/kg anti-TGF-beta antibody or control antibody. Serum creatinine and BUN levels and renal histology were determined. Real time PCR and western analysis were used to quantify mRNA and protein expression. RESULTS: BUN and creatinine were elevated in TAC-treated rats. TAC increased expression of TGF-beta (37-fold) and NADPH oxidase subunits, NOX-1 (18-fold), p22(phox) (31-fold) and Rac-1 mRNA (20-fold), respectively. Contrariwise, expression of antioxidant genes, superoxide dismutase (SOD) and thioredoxin (TRX) was decreased. Anti-TGF-beta antibody but not control antibody reversed the TAC-induced changes in gene expression, renal histology and function. CONCLUSIONS: Our findings suggest a potential for anti-TGF-beta antibody as a novel adjunct therapeutic tool to prevent TAC-induced nephrotoxicity in transplant recipients. The mechanism of protection involves suppression of TGF-beta and the expression of genes that regulate oxidative stress. Moreover, the specific up-regulation of NOX-1, a non-phagocytic NADPH oxidase subunit and its reversal by anti-TGF-beta antibody strongly implicates for the first time the up-regulation of renal parenchymal cell NADPH oxidase in the aetiology of immunosuppression-induced nephrotoxicity. PMID- 17085465 TI - The expression of matrix metalloproteinase-11 protein in various types of glomerulonephritis. AB - BACKGROUND: Matrix metalloproteinases (MMPs) have been implicated to play important roles in a number of pathological processes such as inflammation. In human glomeruli, the mesangial matrix turnover is controlled by a dynamic equilibrium between synthesis and degradation to which metalloproteinases are known to contribute. Metalloproteinase-11 (MMP-11) was originally discovered as a gene whose expression was associated with tissue remodelling. The aim of this study was to investigate whether MMP-11 protein is expressed in various types of glomerulonephritis and to elucidate the role of this expression. METHODS: Using standard immunohistochemistry, we analysed MMP-11 expression in renal biopsies from 95 patients with primary glomerulonephritis (n = 44) and secondary, either lupus-associated glomerulonephritis (n = 22) or pauci-immune, ANCA-associated glomerulonephritis due to small vessel vasculitis (n = 23) or Wegener's granulomatosis (n = 6). The examined cases were divided into two groups (proliferative and non-proliferative). Anti-Ki67 and -CD68 immunostaining was also performed in order to estimate cell proliferation and number of macrophages, respectively. RESULTS: MMP-11 immunopositivity was detected in the glomeruli of the majority of pathological samples. The highest incidence of MMP-11 immunopositivity (26.3%) was noticed in glomerulonephritides associated with microscopic polyangiitis and Wegener's granulomatosis. Generally, MMP-11 was often expressed in segmental areas of sclerosis, microadhesions, cellular and fibrocellular crescents. Fibrotic crescents and fibrotic glomeruli were constantly MMP-11-immunonegative. In MMP-11 immunoreactive glomeruli, increased numbers of macrophages were often detected in the mesangium (P = 0.001), while no such observation could be made with regard to proliferating cells (P = 0.170). CONCLUSIONS: MMP-11, like an inflammatory mediator, may exert a chemotactic influence on macrophages which aggregate in the mesangium; MMP-11 is not likely to have a parallel mitogenic or antifibrotic effect in diseased glomeruli. PMID- 17085466 TI - Vitamin E and selenium co-supplementation attenuates oxidative stress in haemodialysis patients receiving intra-dialysis iron infusion. PMID- 17085467 TI - Distinct expression pattern of IFN-alpha and TNF-alpha in juvenile idiopathic arthritis synovial tissue. AB - OBJECTIVES: Recent laboratory and clinical data suggest that two prototype autoimmune diseases, systemic lupus erythematosus and rheumatoid arthritis are mainly driven by distinct cytokines, interferon (IFN)-alpha and tumour necrosis factor (TNF)-alpha, respectively. We here investigated the presence and characteristics of natural type I IFN-producing cells (IPCs), as well as IFN alpha and TNF-alpha expression at sites of inflammation in juvenile idiopathic arthritis (JIA). METHODS: Peripheral blood (PB) and synovial fluid (SF) mononuclear cells (MNCs) (n = 25 each) from JIA patients with active disease were studied. IPCs were identified as BCDA-2(+)CD123(+)HLA-DR(+)CD45RA(+) cells, and dendritic cells (DCs) as CD11c(+)CD14(-/low)lin(-) cells by flow cytometry. IPCs and DCs were analysed for Toll-like receptor-7 and -9 mRNA expression by real time polymerase chain reaction. IFN-alpha was measured by enzyme-linked immunosorbent assay in serum, SF and in supernatants of influenza virus-infected, cultured IPCs. Synovial tissues of n = 6 additional JIA patients were analysed by immunohistochemistry using mAbs against CD123, IFN-alpha, TNF-alpha, CD3, CD19 and CD138. RESULTS: IPCs were enriched in SF MNCs compared with PB MNCs in all JIA patients. Influenza-induced, but no spontaneous IFN-alpha release was detected from SF IPCs, and serum and SF IFN-alpha levels were not elevated. Nonetheless, in synovial tissue IFN-alpha producing cells accumulated at inflammatory lymph-follicular-like structures, while TNF-alpha producing cells were mostly found at the lining and sublining layers. CONCLUSIONS: These data suggest that besides TNF-alpha-expressing cells, IFN-alpha-producing IPCs are involved in initiation, maintenance or regulation of the inflammatory response in JIA. PMID- 17085468 TI - Patient preferences in choosing anti-TNF therapies-R1. PMID- 17085469 TI - Unilateral polymyalgia rheumatica with controlateral sympathetic dystrophy syndrome. A case of asymmetrical involvement due to pre-existing peripheral palsy. PMID- 17085470 TI - Assessing the effects of land-use change on plant traits, communities and ecosystem functioning in grasslands: a standardized methodology and lessons from an application to 11 European sites. AB - BACKGROUND AND AIMS: A standardized methodology to assess the impacts of land-use changes on vegetation and ecosystem functioning is presented. It assumes that species traits are central to these impacts, and is designed to be applicable in different historical, climatic contexts and local settings. Preliminary results are presented to show its applicability. METHODS: Eleven sites, representative of various types of land-use changes occurring in marginal agro-ecosystems across Europe and Israel, were selected. Climatic data were obtained at the site level; soil data, disturbance and nutrition indices were described at the plot level within sites. Sixteen traits describing plant stature, leaf characteristics and reproductive phase were recorded on the most abundant species of each treatment. These data were combined with species abundance to calculate trait values weighed by the abundance of species in the communities. The ecosystem properties selected were components of above-ground net primary productivity and decomposition of litter. KEY RESULTS: The wide variety of land-use systems that characterize marginal landscapes across Europe was reflected by the different disturbance indices, and were also reflected in soil and/or nutrient availability gradients. The trait toolkit allowed us to describe adequately the functional response of vegetation to land-use changes, but we suggest that some traits (vegetative plant height, stem dry matter content) should be omitted in studies involving mainly herbaceous species. Using the example of the relationship between leaf dry matter content and above-ground dead material, we demonstrate how the data collected may be used to analyse direct effects of climate and land use on ecosystem properties vs. indirect effects via changes in plant traits. CONCLUSIONS: This work shows the applicability of a set of protocols that can be widely applied to assess the impacts of global change drivers on species, communities and ecosystems. PMID- 17085471 TI - Stomatal patchiness and task-performing networks. AB - BACKGROUND: Patchy stomatal conductance is a poorly understood and little-studied phenomenon. It is relatively common, yet it appears to be detrimental to water use efficiency under some conditions and has no immediately obvious physiological function of any kind. Much of the difficulty in studying patchy stomatal conductance is tied to its unpredictability, both in occurrence and in characteristics. SCOPE AND CONCLUSIONS: Statistical analyses of the variability of stomatal patchiness reveal remarkable similarities to structures and behaviours found in locally connected networks of dynamic units that perform tasks. Such systems solve problems that reside at the level of the entire network despite the absence of a central processor or a mechanism for directly sharing information over the entire system. Frequently, task performance is emergent, in the sense that no unit independently performs the task. Because each unit in the network can communicate with only its immediate neighbours, problem solving is accomplished by the states of the individual units self-organizing into synchronized, collective patterns. In some cases, patches of states form and move coherently over the network, thus providing a means for distantly separated parts of the network to communicate. Often, exactly what form these patches take and how they move as the units synchronize is highly unpredictable. In analogy with such networks, it is suggested that stomatal patchiness may be a signature that plants optimize gas exchange in a more sophisticated and adaptive manner than if performed by their individual stomata independently. PMID- 17085472 TI - Floral anatomy of Paepalanthoideae (Eriocaulaceae, Poales) and their Nectariferous structures. AB - BACKGROUND AND AIMS: Eriocaulaceae (Poales) is currently divided in two subfamilies: Eriocauloideae, which comprises two genera and Paepalanthoideae, with nine genera. The floral anatomy of Actinocephalus polyanthus, Leiothrix fluitans, Paepalanthus chlorocephalus, P. flaccidus and Rondonanthus roraimae was studied here. The flowers of these species of Paepalanthoideae are unisexual, and form capitulum-type inflorescences. Staminate and pistillate flowers are randomly distributed in the capitulum and develop centripetally. This work aims to establish a floral nomenclature for the Eriocaulaceae to provide more information about the taxonomy and phylogeny of the family. METHODS: Light microscopy, scanning electron microscopy and chemical tests were used to investigate the floral structures. KEY RESULTS: Staminate and pistillate flowers are trimerous (except in P. flaccidus, which presents dimerous flowers), and the perianth of all species is differentiated into sepals and petals. Staminate flowers present an androecium with scale-like staminodes (not in R. roraimae) and fertile stamens, and nectariferous pistillodes. Pistillate flowers present scale-like staminodes (except for R. roraimae, which presents elongated and vascularized staminodes), and a gynoecium with a hollow style, ramified in stigmatic and nectariferous portions. CONCLUSIONS: The scale-like staminodes present in the species of Paepalanthoideae indicate a probable reduction of the outer whorl of stamens present in species of Eriocauloideae. Among the Paepalanthoideae genera, Rondonanthus, which is probably basal, shows vascularized staminodes in their pistillate flowers. The occurrence of nectariferous pistillodes in staminate flowers and that of nectariferous portions of the style in pistillate flowers of Paepalanthoideae are emphasized as nectariferous structures in Eriocaulaceae. PMID- 17085473 TI - The effects of mechanical stress and spectral shading on the growth and allocation of ten genotypes of a stoloniferous plant. AB - BACKGROUND AND AIMS: Because plants protect each other from wind, stand density affects both the light climate and the amount of mechanical stress experienced by plants. But the potential interactive effects of mechanical stress and canopy shading on plant growth have rarely been investigated and never in stoloniferous plants which, due to their creeping growth form, can be expected to respond differently to these factors than erect plants. METHODS: Plants of ten genotypes of the stoloniferous species Potentilla reptans were subjected to two levels of mechanical stress (0 or 40 daily flexures) and two levels of spectral shading (15 % of daylight with a red:far red ratio of 0.3 vs. 50 % daylight and a red:far red ratio of 1.2). KEY RESULTS: Mechanically stressed plants produced more leaves with shorter more flexible petioles, more roots, and more but less massive stolons. Responses to spectral shading were mostly in the opposite direction to thigmomorphogenesis, including the production of thinner, taller petioles made of more rigid tissue. The degree of thigmomorphogenesis was either independent of light climate or stimulated by spectral shading. At the genotypic level there were no clear correlations between responses to shade and mechanical stress. CONCLUSIONS: These results suggest that in stoloniferous plants mechanical stress results in clones with a more compact, shorter shoot structure and more roots. This response does not appear to be suppressed by canopy shading, which suggests that wind shielding (reduced mechanical stress) by neighbours in dense vegetation serves as a cue that induces shade avoidance responses such as increased petiole elongation. PMID- 17085474 TI - Reproductive biology and mating system estimates of two Andean melocacti, Melocactus schatzlii and M. andinus (Cactaceae). AB - BACKGROUND AND AIMS: The genus Melocactus comprises 36 species of globose cacti with the most derived traits in the Cereeae tribe. It is the proper study system to examine what are the most derived reproductive strategies within that tribe. This study aims to characterize the reproductive biology and to estimate the mating system parameters of two Andean melocacti, Melocactus schatzlii and M. andinus. METHODS: The reproductive attributes of the two species were described, including floral morphology, anthesis patterns, floral rewards, floral visitors and visitation patterns. Levels of self-compatibility and autonomous self pollination were estimated by hand-pollination experiments. Mating system estimates were obtained by conducting progeny array analyses using isozymes. KEY RESULTS: The flowers of the two species present the typical hummingbird pollination syndrome. Despite their morphological resemblance, the two species differ in flower size, pollen and ovule production and anthesis pattern. Their main pollinator agents are hummingbirds, four species in M. schatzlii and one species in M. andinus. Both cacti are self-compatible and capable of self pollination without the aid of pollen vectors. Population-level outcrossing rate was higher for M. schatzlii (t(m)=0.9) than for M. andinus (t(m)=0.4). At the family level, outcrossing rates for most mothers of M. schatzlii were higher (t(m)>0.8) than for M. andinus (t(m)<0.5). CONCLUSIONS: Although the two cacti are capable of selfing, M. schatzlii is a predominantly outcrossing species, while M. andinus behaves as a mixed-mating cactus. Hummingbirds are the only pollinators responsible for outcrossing and gene flow events in these species. In their absence, both melocacti set seeds by selfing. Based on its low population size, restricted distribution in Venezuela, low rates of floral visits, and high levels of inbreeding, M. andinus is considered to be an endangered species deserving further study to define its conservation status. PMID- 17085475 TI - The role of reproductive phenology, seedling emergence and establishment of perennial Salix gordejevii in active sand dune fields. AB - BACKGROUND AND AIMS: The function of sexual reproduction of perennials in restoration of vegetation of active dune fields frequently has been underestimated. The objective of this study was to evaluate the role of sexual reproduction of the perennial Salix gordejevii in the revegetation of active dunes. METHODS: Seedling emergence and establishment of S. gordejevii were examined both in controlled experiments (germination at different burial depths with different watering regimes) and in field observations in three dune slacks. The reproductive phenology and soil seed bank of S. gordejevii, the dynamics of soil moisture, the groundwater table and the landform level of three dune slacks were monitored. KEY RESULTS: Seeds of S. gordejevii began maturation on 1 May, and seed dispersal lasted from 8 May to 20 May. Seeds on the soil surface germinated significantly faster than those buried in soil (P<0.05). Seedling emergence was negatively correlated with landform level. When most seedlings emerged, there was a significantly positive correlation between soil moisture and seedling emergence (P<0.01). Rainfall was negatively correlated with seedling emergence. Seedling establishment was significantly and positively correlated with seedling emergence (P<0.05), and 72.3 % of the emergent seedlings were established at the end of the growing season. These results indicated that (a) seeds matured and dispersed before the rainy season; (b) seeds germinated as soon as they contacted a moist surface and relied more on soil moisture than on rainfall; and (c) more seedlings emerged at lower sampling points in dune slacks. CONCLUSIONS: In natural conditions, restoration of active sand dune fields generally commences with revegetation of dune slacks where sexual reproduction of perennials contributes greatly to species encroachment and colonization and hence plays an important role in restoration of active dune fields. Furthermore, aeolian erosion in dune slacks, leading to good soil moisture, facilitates seed germination, seedling emergence and establishment of S. gordejevii. PMID- 17085476 TI - Above- and below-ground biomass relationships across 1534 forested communities. AB - BACKGROUND AND AIMS: Prior work has shown that above- and below-ground dry biomass across individual plants scale in a near isometric manner across phyletically and ecologically diverse species. Allometric theory predicts that a similar isometric scaling relationship should hold true across diverse forest types, regardless of vegetational composition. METHODS: To test this hypothesis, two compendia for forest-level above- and below-ground dry biomass per hectare (M(A) and M(R), respectively) were examined to test the hypothesis that M(A) vs. M(R) scales isometrically and in the same manner as reported for data from individual plants. Model Type II regression protocols were used to compare the numerical values of M(A) vs. M(R) scaling exponents (i.e. slopes of log-log linear relationships) for the combined data sets (n =1534), each of the two data sets, and data sorted into a total of 17 data subsets for community- and biome types as well as communities dominated by angiosperms or conifers. KEY RESULTS: Among the 20 regressions examined, 15 had scaling exponents that were indistinguishable from that reported for M(A) vs. M(R) across individual plants. The isometric hypothesis could not be strictly rejected on statistical grounds; four of these 15 exponents had broad 95% confidence intervals resulting from small sample sizes. Significant variation was observed in the y-intercepts of the 20 regression curves, because of absolute differences in M(A) or M(R). CONCLUSIONS: The allometries of forest- and individual plant-level M(A) vs. M(R) relationships share strikingly similar scaling exponents, but differ because of considerable variation in y-intercepts. These results support prior allometric theory and provide boundary conditions for the scaling of M(A) and M(R). PMID- 17085477 TI - ZNF366 is an estrogen receptor corepressor that acts through CtBP and histone deacetylases. AB - The regulation of gene expression by estrogen receptor-alpha (ERalpha) requires the coordinated and temporal recruitment of diverse sets of transcriptional co regulator complexes, which mediate nucleosome remodelling and histone modification. Using ERalpha as bait in a yeast two-hybrid screen, we have identified a novel ERalpha-interacting protein, ZNF366, which is a potent corepressor of ERalpha activity. The interaction between ZNF366 and ERalpha has been confirmed in vitro and in vivo, and is mediated by the zinc finger domains of the two proteins. Further, we show that ZNF366 acts as a corepressor by interacting with other known ERalpha corepressors, namely RIP140 and CtBP, to inhibit expression of estrogen-responsive genes in vivo. Together, our results indicate that ZNF366 may play an important role in regulating the expression of genes in response to estrogen. PMID- 17085478 TI - Role of metal ions in catalysis by HIV integrase analyzed using a quantitative PCR disintegration assay. AB - Paired metal ions have been proposed to be central to the catalytic mechanisms of RNase H nucleases, bacterial transposases, Holliday junction resolvases, retroviral integrases and many other enzymes. Here we present a sensitive assay for DNA transesterification in which catalysis by human immunodeficiency virus type 1 (HIV-1) integrase (IN) connects two DNA strands (disintegration reaction), allowing detection using quantitative PCR (qPCR). We present evidence suggesting that the three acidic residues of the IN active site function through metal binding using metal rescue. In this method, the catalytic acidic residues were each substituted with cysteines. Mn2+ binds tightly to the sulfur atoms of the cysteine residues, but Mg2+ does not. We found that Mn2+, but not Mg2+, could rescue catalysis of each cysteine-substituted enzyme, providing evidence for functionally important metal binding by all three residues. We also used the PCR boosted assay to show that HIV-1 IN could carry out transesterification reactions involving DNA 5' hydroxyl groups as well as 3' hydroxyls as nucleophiles. Lastly, we show that Mn2+ by itself (i.e. without enzyme) can catalyze formation of a low level of PCR-amplifiable product under extreme conditions, allowing us to estimate the rate enhancement due to the IN-protein scaffold as at least 60 million-fold. PMID- 17085479 TI - Patome: a database server for biological sequence annotation and analysis in issued patents and published patent applications. AB - With the advent of automated and high-throughput techniques, the number of patent applications containing biological sequences has been increasing rapidly. However, they have attracted relatively little attention compared to other sequence resources. We have built a database server called Patome, which contains biological sequence data disclosed in patents and published applications, as well as their analysis information. The analysis is divided into two steps. The first is an annotation step in which the disclosed sequences were annotated with RefSeq database. The second is an association step where the sequences were linked to Entrez Gene, OMIM and GO databases, and their results were saved as a gene-patent table. From the analysis, we found that 55% of human genes were associated with patenting. The gene-patent table can be used to identify whether a particular gene or disease is related to patenting. Patome is available at http://www.patome.org/; the information is updated bimonthly. PMID- 17085480 TI - DTL/CDT2 is essential for both CDT1 regulation and the early G2/M checkpoint. AB - Checkpoint genes maintain genomic stability by arresting cells after DNA damage. Many of these genes also control cell cycle events in unperturbed cells. By conducting a screen for checkpoint genes in zebrafish, we found that dtl/cdt2 is an essential component of the early, radiation-induced G2/M checkpoint. We subsequently found that dtl/cdt2 is required for normal cell cycle control, primarily to prevent rereplication. Both the checkpoint and replication roles are conserved in human DTL. Our data indicate that the rereplication reflects a requirement for DTL in regulating CDT1, a protein required for prereplication complex formation. CDT1 is degraded in S phase to prevent rereplication, and following DNA damage to prevent origin firing. We show that DTL associates with the CUL4-DDB1 E3 ubiquitin ligase and is required for CDT1 down-regulation in unperturbed cells and following DNA damage. The cell cycle defects of Dtl deficient zebrafish are suppressed by reducing Cdt1 levels. In contrast, the early G2/M checkpoint defect appears to be Cdt1-independent. Thus, DTL promotes genomic stability through two distinct mechanisms. First, it is an essential component of the CUL4-DDB1 complex that controls CDT1 levels, thereby preventing rereplication. Second, it is required for the early G2/M checkpoint. PMID- 17085481 TI - Endotoxic shock in AUF1 knockout mice mediated by failure to degrade proinflammatory cytokine mRNAs. AB - Excessive production of proinflammatory cytokines, particularly tumor necrosis factor-alpha (TNFalpha) and interleukin-1beta (IL-1beta), plays a critical role in septic shock induced by bacterial endotoxin (endotoxemia). Precise control of cytokine expression depends on rapid degradation of cytokine mRNAs, mediated by an AU-rich element (ARE) in the 3' noncoding region and by interacting ARE binding proteins, which control the systemic inflammatory response. To understand the function of the ARE-binding protein AUF1, we developed an AUF1 knockout mouse. We show that AUF1 normally functions to protect against the lethal progression of endotoxemia. Upon endotoxin challenge, AUF1 knockout mice display symptoms of severe endotoxic shock, including vascular hemorrhage, intravascular coagulation, and high mortality, resulting from overproduction of TNFalpha and IL 1beta. Overexpression of these two cytokines is specific, and shown to result from an inability to rapidly degrade these mRNAs in macrophages following induction. Neutralizing antibodies to TNFalpha and IL-1beta protect AUF1 knockout mice against lethal endotoxic shock. These and other data describe a novel post transcriptional mechanism whereby AUF1 acts as a crucial attenuator of the inflammatory response, promoting the rapid decay of selective proinflammatory cytokine mRNAs following endotoxin activation. Defects in the AUF1 post transcriptionally controlled pathway may be involved in human inflammatory disease. PMID- 17085482 TI - Direct interaction between DNMT1 and G9a coordinates DNA and histone methylation during replication. AB - Chromatin methylation is necessary for stable repression of gene expression during mammalian development. During cell division, DNMT1 maintains the DNA methylation pattern of the newly synthesized daughter strand, while G9a methylates H3K9. Here, DNMT1 is shown to directly bind G9a both in vivo and in vitro and to colocalize in the nucleus during DNA replication. The complex of DNMT1 and G9a colocalizes with dimethylated H3K9 (H3K9me2) at replication foci. Similarly, another H3K9 histone methyltransferase, SUV39H1, colocalizes with DNMT1 on heterochromatic regions of the nucleoli exclusively before cell division. Both DNMT1 and G9a are loaded onto the chromatin simultaneously in a ternary complex with loading factor PCNA during chromatin replication. Small interfering RNA (siRNA) knockdown of DNMT1 impairs DNA methylation, G9a loading, and H3K9 methylation on chromatin and rDNA repeats, confirming DNMT1 as the primary loading factor. Additionally, the complex of DNMT1 and G9a led to enhanced DNA and histone methylation of in vitro assembled chromatin substrates. Thus, direct cooperation between DNMT1 and G9a provides a mechanism of coordinated DNA and H3K9 methylation during cell division. PMID- 17085483 TI - Haplotype-specific expression of exon 10 at the human MAPT locus. AB - Neurofibrillary tangles composed of exon 10+ microtubule associated protein tau (MAPT) deposits are the characteristic feature of the neurodegenerative diseases progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD). PSP, CBD and more recently Alzheimer's disease and Parkinson's disease, are associated with the MAPT H1 haplotype, but the relationship between genotype and disease remains unclear. Here, we investigate the hypothesis that H1 expresses more exon 10+ MAPT mRNA compared to the other haplotype, H2, leading to a greater susceptibility to neurodegeneration in H1 carriers. We performed allele-specific gene expression on two H1/H2 heterozygous human neuronal cell lines, and 14 H1/H2 heterozygous control individual post-mortem brain tissue from two brain regions. In both tissue culture and post-mortem brain tissue, we show that the MAPT H1 haplotype expresses significantly more exon 10+ MAPT mRNA than H2. In post-mortem brain tissue, we show that the total level of MAPT expression from H1 and H2 is not significantly different, but that the H1 chromosome expresses up to 1.43-fold more exon 10+ MAPT mRNA than H2 in the globus pallidus, a brain region highly affected by tauopathy (maximum exon 10+ MAPT H1:H2 transcript ratio=1.425, SD=0.205, P<0.0001), and up to 1.29-fold more exon 10+ MAPT mRNA than H2 in the frontal cortex (maximum exon 10+ MAPT H1:H2 transcript ratio=1.291, SD=0.315, P=0.006). These data may explain the increased susceptibility of H1 carriers to neurodegeneration and suggest a potential mechanism between MAPT genetic variability and the pathogenesis of neurodegenerative disease. PMID- 17085484 TI - Haplotype spanning TTC12 and ANKK1, flanked by the DRD2 and NCAM1 loci, is strongly associated to nicotine dependence in two distinct American populations. AB - Nicotine dependence (ND) is a moderately heritable trait. We ascertained a set of 1615 subjects in 632 families [319 African-American (AA) and 313 European American (EA)] based on affected sibling pairs with cocaine or opioid dependence. Subjects were interviewed with the Semi-Structured Assessment for Drug Dependence and Alcoholism (SSADDA). Previously, we identified a modest linkage peak (LOD score =1.97) for ND in the EA part of the sample on chromosome 11q23, a region that includes the NCAM1-TTC12-ANKK1-DRD2 gene cluster. DRD2 and NCAM1 are functional candidate genes for substance dependence; the TTC12 and ANKK1 loci are not well characterized. We genotyped a set of 43 single nucleotide polymorphisms (SNPs) spanning this region, and performed family-based association and haplotype analysis. There was relatively weak evidence for association of the flanking DRD2 and NCAM1 markers to ND, but very strong evidence of association of multiple SNPs at TTC12 and ANKK1 in both populations (minimal P=0.0007 in AAs and minimal P=0.00009 in EAs), and in the pooled sample, as well as strong evidence for highly significant association of a single haplotype spanning TTC12 and ANKK1 to ND in the pooled sample (P=0.0000001). We conclude that a risk locus for ND, important both in AAs and EAs, maps to a region that spans TTC12 and ANKK1. Functional studies of these loci are warranted. These results provide additional information useful in evaluating the many earlier discrepant findings regarding association of DRD2 with substance dependence. PMID- 17085485 TI - The Annexin code: revealing endocarditis. PMID- 17085486 TI - Analysis of two-dimensional dissociation constant of laterally mobile cell adhesion molecules. AB - We formulate a general analysis to determine the two-dimensional dissociation constant (2D Kd), and use this method to study the interaction of CD2-expressing T cells with glass-supported planar bilayers containing fluorescently labeled CD58, a CD2 counter-receptor. Both CD2 and CD58 are laterally mobile in their respective membranes. Adhesion is indicated by accumulation of CD2 and CD58 in the cell-bilayer contact area; adhesion molecule density and contact area size attain equilibrium within 40 min. The standard (Scatchard) analysis of solution phase binding is not applicable to the case of laterally mobile adhesion molecules due to the dynamic nature of the interaction. We derive a new binding equation, B/F=[(Ntxf)/(KdxScell)]-[(Bxp)/Kd], where B and F are bound and free CD58 density in the contact area, respectively; Nt is CD2 molecule number per cell; f is CD2 fractional mobility; Scell is cell surface area; and p is the ratio of contact area at equilibrium to Scell. We use this analysis to determine that the 2D Kd for CD2-CD58 is 5.4-7.6 molecules/microm2. 2D Kd analysis provides a general and quantitative measure of the mechanisms regulating cell-cell adhesion. PMID- 17085487 TI - NMR simulation analysis of statistical effects on quantifying cerebrovascular parameters. AB - Determining tissue structure and composition from the behavior of the NMR transverse relaxation during free induction decay and spin echo formation has seen significant advances in recent years. In particular, the ability to quantify cerebrovascular network parameters such as blood volume and deoxyhemoglobin concentration from the NMR signal dephasing has seen intense focus. Analytical models have been described, based on statistical averaging of randomly oriented cylinders in both the static and slow diffusion regimes. However, the error in estimates obtained from these models when applied to systems in which the statistical assumptions of many, randomly oriented perturbers are violated has not been systematically investigated. Using a deterministic simulation that can include diffusion, we find that the error in estimated venous blood volume fraction and deoxyhemoglobin concentration obtained using a static dephasing regime statistical model is inversely related to the square root of number of blood vessels. The most important implication of this is that the minimum imaging resolution for accurate deoxyhemoglobin and blood volume estimation is not bound by hardware limitations, but rather by the underlying tissue structure. PMID- 17085488 TI - Receptor-mediated and intrinsic polarization and their interaction in chemotaxing cells. AB - Polarization--the clear and persistent localization of different signaling molecules to opposite ends of the cell-is critical for effective chemotaxis in eukaryotic systems. In many systems, polarization can also occur without an externally imposed chemical gradient. We build a modeling framework to study the relationship between the intrinsic capacity for polarization, and that induced by an external gradient. Working within this framework, we analyze different scenarios for the interaction of these pathways. The models are qualitatively simplified, motivated by known properties of the signaling pathways. We also examine the possible role of nonlinear transitions occurring in the polarization pathways. The modeling framework generates testable predictions regarding the relationship between intrinsic polarization and that induced during chemotaxis, and is the first step toward a systematic analysis of the interaction between these pathways. PMID- 17085489 TI - Spectral methods for parametric sensitivity in stochastic dynamical systems. AB - Stochastic dynamical systems governed by the chemical master equation find use in the modeling of biological phenomena in cells, where they provide more accurate representations than their deterministic counterparts, particularly when the levels of molecular population are small. The analysis of parametric sensitivity in such systems requires appropriate methods to capture the sensitivity of the system dynamics with respect to variations of the parameters amid the noise from inherent internal stochastic effects. We use spectral polynomial chaos expansions to represent statistics of the system dynamics as polynomial functions of the model parameters. These expansions capture the nonlinear behavior of the system statistics as a result of finite-sized parametric perturbations. We obtain the normalized sensitivity coefficients by taking the derivative of this functional representation with respect to the parameters. We apply this method in two stochastic dynamical systems exhibiting bimodal behavior, including a biologically relevant viral infection model. PMID- 17085490 TI - Adhesive dynamics simulations of the shear threshold effect for leukocytes. AB - Many experiments have measured the effect of force on the dissociation of single selectin bonds, but it is not yet clear how the force dependence of molecular dissociation can influence the rolling of cells expressing selectin molecules. Recent experiments using constant-force atomic force microscopy or high resolution microscopic observations of pause-time distributions of cells in a flow chamber show that for some bonds, the dissociation rate is high at low force and initially decreases with force, indicating a catch bond. As the force continues to increase, the dissociation rate increases again, like a slip bond. It has been proposed that this catch-slip bond leads to the shear threshold effect, in which a certain level of shear rate is required to achieve rolling. We have incorporated a catch-slip dissociation rate into adhesive dynamics simulations of cell rolling. Using a relatively simple model for the shear controlled association rate for selectin bonds, we were able to recreate characteristics of the shear threshold effect seen most prominently for rolling through L-selectin. The rolling velocity as a function of shear rate showed a minimum near 100 s-1. Furthermore, cells were observed to roll at a shear rate near the threshold, but detach and move more quickly when the shear rate was dropped below the threshold. Finally, using adhesive dynamics, we were able to determine ranges of parameters necessary to see the shear threshold effect in the rolling velocity. In summary, we found through simulation that the catch-slip behavior of selectin bonds can be responsible for the shear threshold effect. PMID- 17085491 TI - On the lower susceptibility of oseltamivir to influenza neuraminidase subtype N1 than those in N2 and N9. AB - Aiming to understand, at the molecular level, why oseltamivir (OTV) cannot be used for inhibition of human influenza neuraminidase subtype N1 as effectively as for subtypes N2 and N9, molecular dynamics simulations were carried out for the three complexes, OTV-N1, OTV-N2, and OTV-N9. The three-dimensional OTV-N2 and OTV N9 initial structures were represented by the x-ray structures, whereas that of OTV-N1, whose x-ray structure is not yet solved, was built up using the aligned sequence of H5N1 isolated from humans in Thailand with the x-ray structure of the N2-substrate as the template. In comparison to the OTV-N2 and OTV-N9 complexes, dramatic changes were observed in the OTV conformation in the OTV-N1 complex in which two of its bulky side chains, N-acethyl (-NHAc) and 1-ethylproxy group ( OCHEt2), were rotated to adjust the size to fit into the N1 catalytic site. This change leads directly to the rearrangements of the OTV's environment, which are i), distances to its neighbors, W-178 and E-227, are shorter whereas those to residues R-224, E-276, and E-292 are longer; ii), hydrogen bonds to the two nearest neighbors, R-224 and E-276, are still conserved in distance and number as well as percentage occupation; iii), the calculated ligand/enzyme binding free energies of -7.20, -13.44, and -13.29 kcal/mol agree with their inhibitory activities in terms of the experimental IC50 of 36.1-53.2 nM, 1.9-2.7 nM, and 9.5 17.7 nM for the OTV-N1, OTV-N2, and OTV-N9 complexes, respectively; and iv), hydrogen-bond breaking and creation between the OTV and neighborhood residues are accordingly in agreement with the ligand solvation/desolvation taking place in the catalytic site. PMID- 17085492 TI - Quantitative characterization of biological liquids for third-harmonic generation microscopy. AB - Third-harmonic generation (THG) microscopy provides images of unstained biological samples based on spatial variations in third-order nonlinear susceptibility, refractive index, and dispersion. In this study, we establish quantitative values for the third-order nonlinear susceptibilities of several solvents (water, ethanol, glycerol), physiological aqueous (ions, amino acids, polypeptides, bovine serum albumin, glucose) and lipid (triglycerides, cholesterol) solutions as a function of solute concentration in the 1.05-1.25 microm excitation range. We use these data in conjunction with imaging experiments to show that THG imaging with approximately 1.2 microm excitation lacks specificity and sensitivity to detect physiological ion concentration changes, and that nonaqueous structures such as lipid bodies provide a more robust source of signal. Finally, we illustrate the impact of index-matching liquids in THG images. These data provide a basis for interpreting biological THG images and for developing additional applications. PMID- 17085493 TI - Structure and molecular mechanism of Bacillus anthracis cofactor-independent phosphoglycerate mutase: a crucial enzyme for spores and growing cells of Bacillus species. AB - Phosphoglycerate mutases (PGMs) catalyze the isomerization of 2- and 3 phosphoglycerates and are essential for glucose metabolism in most organisms. This study reports the production, structure, and molecular dynamics analysis of Bacillus anthracis cofactor-independent PGM (iPGM). The three-dimensional structure of B. anthracis PGM is composed of two structural and functional domains, the phosphatase and transferase. The structural relationship between these two domains is different than in the B. stearothermophilus iPGM structure determined previously. However, the structures of the two domains of B. anthracis iPGM show a high degree of similarity to those in B. stearothermophilus iPGM. The novel domain arrangement in B. anthracis iPGM and the dynamic property of these domains is directly linked to the mechanism of enzyme catalysis, in which substrate binding is proposed to result in close association of the two domains. The structure of B. anthracis iPGM and the molecular dynamics of this structure provide unique insight into the mechanism of iPGM catalysis, in particular the roles of changes in coordination geometry of the enzyme's two bivalent metal ions and the regulation of this enzyme's activity by changes in intracellular pH during spore formation and germination in Bacillus species. PMID- 17085494 TI - Spectrin domains lose cooperativity in forced unfolding. AB - Spectrin is a multidomain cytoskeletal protein, the component three-helix bundle domains are expected to experience mechanical force in vivo. In thermodynamic and kinetic studies, neighboring domains of chicken brain alpha-spectrin R16 and R17 have been shown to behave cooperatively. Is this cooperativity maintained under force? The effect of force on these spectrin domains was investigated using atomic force microscopy. The response of the individual domains to force was compared to that of the tandem repeat R1617. Importantly, nonhelical linkers (all beta immunoglobulin domains) were used to avoid formation of nonnative helical linkers. We show that, in contrast to previous studies on spectrin repeats, only 3% of R1617 unfolding events gave an increase in contour length consistent with cooperative two-domain unfolding events. Furthermore, the unfolding forces for R1617 were the same as those for the unfolding of R16 or R17 alone. This is a strong indication that the cooperative unfolding behavior observed in the stopped flow studies is absent between these spectrin domains when force is acting as a denaturant. Our evidence suggests that the rare double unfolding events result from misfolding between adjacent repeats. We suggest that this switch from cooperative to independent behavior allows multidomain proteins to maintain integrity under applied force. PMID- 17085495 TI - Self-assembling of peptide/membrane complexes by atomistic molecular dynamics simulations. AB - Model biological membranes consisting of peptide/lipid-bilayer complexes can nowadays be studied by classical molecular dynamics (MD) simulations at atomic detail. In most cases, the simulation starts with an assumed state of a peptide in a preformed bilayer, from which equilibrium configurations are difficult to obtain due to a relatively slow molecular diffusion. As an alternative, we propose an extension of reported work on the self-organization of unordered lipids into bilayers, consisting of including a peptide molecule in the initial random configuration to obtain a membrane-bound peptide simultaneous to the formation of the lipid bilayer. This strategy takes advantage of the fast reorganization of lipids, among themselves and around the peptide, in an aqueous environment. Model peptides of different hydrophobicity, CH3-CO-W2L18W2-NH2 (WL22) and CH3-CO-W2A18W2-NH2 (WA22), in dipalmitoyl-phosphatidylcholine (DPPC) are used as test cases. In the equilibrium states of the peptide/membrane complexes, achieved in time ranges of 50-100 ns, the two peptides behave as expected from experimental and theoretical studies. The strongly hydrophobic WL22 is inserted in a transmembrane configuration and the marginally apolar, alanine based WA22 is found in two alternative states: transmembrane inserted or parallel to the membrane plane, embedded close to the bilayer interface, with similar stability. This shows that the spontaneous assembly of peptides and lipids is an unbiased and reliable strategy to produce and study models of equilibrated peptide/lipid complexes of unknown membrane-binding mode and topology. PMID- 17085496 TI - Particle image correlation spectroscopy (PICS): retrieving nanometer-scale correlations from high-density single-molecule position data. AB - A new data analysis tool that resolves correlations on the nanometer length and millisecond timescale is derived. This tool, adapted from methods of spatiotemporal image correlation spectroscopy, exploits the high positional accuracy of single-particle tracking. While conventional tracking methods break down if multiple particle trajectories intersect, our method works in principle for arbitrarily large molecule densities and diffusion coefficients as long as individual molecules can be identified. The method is computationally cheap and robust and requires no a priori knowledge about the dynamical coefficients, as opposed to other methods. We demonstrate the validity of the method by Monte Carlo simulations and by application to single-molecule tracking data of membrane anchored proteins in live cells. The results faithfully reproduce those obtained by conventional tracking. Upon activation, a fraction of the small GTPase H-Ras is confined to domains of <200 nm diameter, which further substantiates the prediction that membrane organization is a determinant in cellular signaling. PMID- 17085497 TI - Temperature effects on DNA chip experiments from surface plasmon resonance imaging: isotherms and melting curves. AB - We present an analysis of hybridization experiments on a DNA chip studied by surface plasmon resonance imaging. The reaction constants at various temperatures and for different probe lengths are obtained from Langmuir isotherms and hybridization kinetics. The melting curves from temperature scans are also obtained without any labeling of the targets. The effects of the probe length on the hybridization thermodynamics, deduced from the temperature dependence of the reaction constants as well as from the melting curves, suggest dispersion in the length of the hybridization segments of the probes accessible to the targets. Those are, however, sufficient to suggest efficient point mutation detection from temperature scans. PMID- 17085498 TI - Localized functional chemical stimulation of TE 671 cells cultured on nanoporous membrane by calcein and acetylcholine. AB - Acetylcholine sensitive TE 671 cells were cultured on nanoporous membranes and chemically stimulated by localized application of i), calcein-AM and ii), acetylcholine, respectively, onto the bottom face of the membrane employing an ink jet print head. Stimulus correlated response of cells was recorded by fluorescence microscopy with temporal and spatial resolution. Calcein fluorescence develops as a result of intracellular enzymatic conversion of calcein-AM, whereas Ca(2+) imaging using fluo-4 dye was employed to visualize cellular response to acetylcholine stimulation. Using 25 pl droplets and substance concentration ranging from 10 microM to 1 mM on Nucleopore membranes with pore diameters between 50 nm and 1 microm, a resolution on the order of 50 microm was achieved. PMID- 17085499 TI - Diffusion analysis within single nanometric apertures reveals the ultrafine cell membrane organization. AB - We describe the development of a new methodology to probe the plasma membrane organization of living cells at the nanometric scale. Single nanometric apertures in a metallic film limit the observed membrane area below the optical diffraction barrier. The new approach performs fluorescence correlation spectroscopy with increasing aperture sizes and extracts information on the diffusion process from the whole set of data. In particular, transient diffusion regimes are clearly observed when the probed area comes close to the size of the confining structures. First, this strategy allows identification of the mechanism controlling the diffusion of various fluorescent lipid analogs and green fluorescent protein-tagged proteins. Second, it gives an estimate of the characteristic size of the nanometric membrane heterogeneities, allowing a quantitative study of membrane domains such as lipid rafts. Compared to other optical techniques, this method combines the advantages of high spatio-temporal resolution and direct statistical analysis. PMID- 17085500 TI - Understanding intracellular transport processes pertinent to synthetic gene delivery via stochastic simulations and sensitivity analyses. AB - A major challenge in synthetic gene delivery is to quantitatively predict the optimal design of polymer-based gene carriers (polyplexes). Here, we report a consistent, integrated, and fundamentally grounded computational methodology to address this challenge. This is achieved by accurately representing the spatio temporal dynamics of intracellular structures and by describing the interactions between gene carriers and cellular components at a discrete, nanoscale level. This enables the applications of systems tools such as optimization and sensitivity analysis to search for the best combination of systems parameters. We validate the approach using DNA delivery by polyethylenimine as an example. We show that the cell topology (e.g., size, circularity, and dimensionality) strongly influences the spatiotemporal distribution of gene carriers, and consequently, their optimal intracellular pathways. The model shows that there exists an upper limit on polyplexes' intracellular delivery efficiency due to their inability to protect DNA until nuclear entry. The model predicts that even for optimally designed polyethylenimine vectors, only approximately 1% of total DNA is delivered to the nucleus. Based on comparison with gene delivery by viruses, the model suggests possible strategies to significantly improve transfection efficiencies of synthetic gene vectors. PMID- 17085501 TI - Membrane assembly of simple helix homo-oligomers studied via molecular dynamics simulations. AB - The assembly of simple transmembrane helix homo-oligomers is studied by combining a generalized Born implicit membrane model with replica exchange molecular dynamics simulations to sample the conformational space of various oligomerization states and the native oligomeric conformation. Our approach is applied to predict the structures of transmembrane helices of three proteins- glycophorin A, the M2 proton channel, and phospholamban--using only peptide sequence and the native oligomerization state information. In every case, the methodology reproduces native conformations that are in good agreement with available experimental structural data. Thus, our method should be useful in the prediction of native structures of transmembrane domains of other peptides. When we ignore the experimental constraint on the native oligomerization state and attempt de novo prediction of the structure and oligomerization state based only on sequence and simple energetic considerations, we identify the pentamer as the most stable oligomer for phospholamban. However, for the glycophorin A and the M2 proton channels, we tend to predict higher oligomers as more stable. Our studies demonstrate that reliable predictions of the structure of transmembrane helical oligomers can be achieved when the observed oligomerization state is imposed as a constraint, but that further efforts are needed for the de novo prediction of both structure and oligomeric state. PMID- 17085502 TI - Optical study of DNA surface hybridization reveals DNA surface density as a key parameter for microarray hybridization kinetics. AB - We investigate the kinetics of DNA hybridization reactions on glass substrates, where one 22 mer strand (bound-DNA) is immobilized via phenylene-diisothiocyanate linker molecule on the substrate, the dye-labeled (Cy3) complementary strand (free-DNA) is in solution in a reaction chamber. We use total internal reflection fluorescence for surface detection of hybridization. As a new feature we perform a simultaneous real-time measurement of the change of free-DNA concentration in bulk parallel to the total internal reflection fluorescence measurement. We observe that the free-DNA concentration decreases considerably during hybridization. We show how the standard Langmuir kinetics needs to be extended to take into account the change in bulk concentration and explain our experimental results. Connecting both measurements we can estimate the surface density of accessible, immobilized bound-DNA. We discuss the implications with respect to DNA microarray detection. PMID- 17085503 TI - Mechanism for intein C-terminal cleavage: a proposal from quantum mechanical calculations. AB - Inteins are autocatalytic protein cleavage and splicing elements. A cysteine to alanine mutation at the N-terminal of inteins inhibits splicing and isolates the C-terminal cleavage reaction. Experiments indicate an enhanced C-terminal cleavage reaction rate upon decreasing the solution pH for the cleavage mutant, which cannot be explained by the existing mechanistic framework. We use intein crystal structure data and the information about conserved amino acids to perform semiempirical PM3 calculations followed by high-level density functional theory calculations in both gas phase and implicit solvent environments. Based on these calculations, we propose a detailed "low pH" mechanism for intein C-terminal cleavage. Water plays an important role in the proposed reaction mechanism, acting as an acid as well as a base. The protonation of the scissile peptide bond nitrogen by a hydronium ion is an important first step in the reaction. That step is followed by the attack of the C-terminal asparagine side chain on its carbonyl carbon, causing succinimide formation and simultaneous peptide bond cleavage. The computed reaction energy barrier in the gas phase is approximately 33 kcal/mol and reduces to approximately 25 kcal/mol in solution, close to the 21 kcal/mol experimentally observed at pH 6.0. This mechanism is consistent with the observed increase in C-terminal cleavage activity at low pH for the cleavage mutant of the Mycobacterium tuberculosis RecA mini-intein. PMID- 17085504 TI - The cPLA2 C2alpha domain in solution: structure and dynamics of its Ca2+ activated and cation-free states. AB - Cytosolic phospholipase A2 is involved in several signal transduction pathways where it catalyses release of arachidonic acid from intracellular lipid membranes. Its membrane insertion is facilitated by its independently folding C2alpha domain, which is activated by the binding of two intracellular Ca2+ ions. However, the details of its membrane-insertion mechanism, including its Ca2+ activation mechanism, are not understood. There are several unresolved issues, including the following. There are two experimentally resolved structures of the Ca2+-activated state of its isolated C2alpha domain, one determined using x-ray crystallography and the other determined using NMR spectroscopy, which differ from each other significantly in the spatial region that inserts into the membrane. This by itself adds to ambiguities associated with investigations targeting its mechanism of membrane insertion. Furthermore, there is no experimentally determined structure of its cation-free state, which hinders investigations associated with its cation-activation mechanism. In this work, we generate several unrestrained molecular dynamics trajectories of its isolated C2alpha domain in solution (equivalent to approximately 60 ns) and investigate these issues. Our main results are as follows: a), the Ca2+ coordination scheme of the domain is consistent with the x-ray structure and with previous mutagenesis studies; b), the helical segment of the Ca2+-binding loop, CBL-I, undergoes nanosecond timescale flexing (but not an unwinding), as can be inferred from physiological temperature NMR data and in contrast to low temperature x-ray data; and c), removal of the two activating Ca2+ ions from their binding pockets does not alter the backbone structure of the domain, a result consistent with electron paramagnetic resonance data. PMID- 17085505 TI - A computational study of the protein-ligand interactions in CDK2 inhibitors: using quantum mechanics/molecular mechanics interaction energy as a predictor of the biological activity. AB - We report a combined quantum mechanics/molecular mechanics (QM/MM) study to determine the protein-ligand interaction energy between CDK2 (cyclin-dependent kinase 2) and five inhibitors with the N(2)-substituted 6-cyclohexyl-methoxy purine scaffold. The computational results in this work show that the QM/MM interaction energy is strongly correlated to the biological activity and can be used as a predictor, at least within a family of substrates. A detailed analysis of the protein-ligand structures obtained from molecular dynamics simulations shows specific interactions within the active site that, in some cases, have not been reported before to our knowledge. The computed interaction energy gauges the strength of protein-ligand interactions. Finally, energy decomposition and multiple regression analyses were performed to check the contribution of the electrostatic and van der Waals energies to the total interaction energy and to show the capabilities of the computational model to identify new potent inhibitors. PMID- 17085506 TI - A heat-inducible transcription factor, HsfA2, is required for extension of acquired thermotolerance in Arabidopsis. AB - The expression of heat shock proteins (Hsps) induced by nonlethal heat treatment confers acquired thermotolerance (AT) to organisms against subsequent challenges of otherwise lethal temperature. After the stress signal is removed, AT gradually decays, with decreased Hsps during recovery. AT of sufficient duration is critical for sessile organisms such as plants to survive repeated heat stress in their environment, but little is known regarding its regulation. To identify potential regulatory components, we took a reverse genetics approach by screening for Arabidopsis (Arabidopsis thaliana) T-DNA insertion mutants that show decreased thermotolerance after a long recovery (2 d) under nonstress conditions following an acclimation heat treatment. Among the tested mutants corresponding to 48 heat-induced genes, only the heat shock transcription factor HsfA2 knockout mutant showed an obvious phenotype. Following pretreatment at 37 degrees C, the mutant line was more sensitive to severe heat stress than the wild type after long but not short recovery periods, and this could be complemented by the introduction of a wild-type copy of the HsfA2 gene. Quantitative hypocotyl elongation assay also revealed that AT decayed faster in the absence of HsfA2. Significant reduction in the transcript levels of several highly heat-inducible genes was observed in HsfA2 knockout plants after 4 h recovery or 2 h prolonged heat stress. Immunoblot analysis showed that Hsa32 and class I small Hsp were less abundant in the mutant than in the wild type after long recovery. Our results suggest that HsfA2 as a heat-inducible transactivator sustains the expression of Hsp genes and extends the duration of AT in Arabidopsis. PMID- 17085507 TI - Dissection of the AtNRT2.1:AtNRT2.2 inducible high-affinity nitrate transporter gene cluster. AB - Using a new Arabidopsis (Arabidopsis thaliana) mutant (Atnrt2.1-nrt2.2) we confirm that concomitant disruption of NRT2.1 and NRT2.2 reduces inducible high affinity transport system (IHATS) by up to 80%, whereas the constitutive high affinity transport system (CHATS) was reduced by 30%. Nitrate influx via the low affinity transport system (LATS) was unaffected. Shoot-to-root ratios were significantly reduced compared to wild-type plants, the major effect being upon shoot growth. In another mutant uniquely disrupted in NRT2.1 (Atnrt2.1), IHATS was reduced by up to 72%, whereas neither the CHATS nor the LATS fluxes were significantly reduced. Disruption of NRT2.1 in Atnrt2.1 caused a consistent and significant reduction of shoot-to-root ratios. IHATS influx and shoot-to-root ratios were restored to wild-type values when Atnrt2.1-nrt2.2 was transformed with a NRT2.1 cDNA isolated from Arabidopsis. Disruption of NRT2.2 in Atnrt2.2 reduced IHATS by 19% and this reduction was statistically significant only at 6 h after resupply of nitrate to nitrogen-deprived plants. Atnrt2.2 showed no significant reduction of CHATS, LATS, or shoot-to-root ratios. These results define NRT2.1 as the major contributor to IHATS. Nevertheless, when maintained on agar containing 0.25 mm KNO(3) as the sole nitrogen source, Atnrt2.1-nrt2.2 consistently exhibited greater stress and growth reduction than Atnrt2.1. Evidence from real-time PCR revealed that NRT2.2 transcript abundance was increased almost 3-fold in Atnrt2.1. These findings suggest that NRT2.2 normally makes only a small contribution to IHATS, but when NRT2.1 is lost, this contribution increases, resulting in a partial compensation. PMID- 17085508 TI - Genome-wide analysis of the Arabidopsis leaf transcriptome reveals interaction of phosphate and sugar metabolism. AB - Global gene expression was analyzed in Arabidopsis (Arabidopsis thaliana) by microarrays comprising 21,500 genes. Leaf segments derived from phosphorus (P) starved and P-replenished plants were incubated with or without sucrose (Suc) to obtain tissues with contrasting combinations of P and carbohydrate levels. Transcript profiling revealed the influence of the two factors individually and the interactions between P- and sugar-dependent gene regulation. A large number of gene transcripts changed more than 2-fold: In response to P starvation, 171 genes were induced and 16 repressed, whereas Suc incubation resulted in 337 induced and 307 repressed genes. A number of new candidate genes involved in P acquisition were discovered. In addition, several putative transcription factors and signaling proteins of P sensing were disclosed. Several genes previously identified to be sugar responsive were also regulated by P starvation and known P responsive genes were sugar inducible. Nearly 150 genes were synergistically or antagonistically regulated by the two factors. These genes exhibit more prominent or contrasting regulation in response to Suc and P in combination than expected from the effect of the two factors individually. The genes exhibiting interactions form three main clusters with different response patterns and functionality of genes. One cluster (cluster 1) most likely represents a regulatory program to support increased growth and development when both P and carbohydrates are ample. Another cluster (cluster 3) represents genes induced to alleviate P starvation and these are further induced by carbohydrate accumulation. Thus, interactions between P and Suc reveal two different signaling programs and novel interactions in gene regulation in response to environmental factors. cis-Regulatory elements were analyzed for each factor and for interaction clusters. PHR1 binding sites were more frequent in promoters of P regulated genes as compared to the entire Arabidopsis genome, and E2F and PHR1 binding sites were more frequent in interaction clusters 1 and 3, respectively. PMID- 17085509 TI - A Phytophthora infestans cystatin-like protein targets a novel tomato papain-like apoplastic protease. AB - There is emerging evidence that the proteolytic machinery of plants plays important roles in defense against pathogens. The oomycete pathogen Phytophthora infestans, the agent of the devastating late blight disease of tomato (Lycopersicon esculentum) and potato (Solanum tuberosum), has evolved an arsenal of protease inhibitors to overcome the action of host proteases. Previously, we described a family of 14 Kazal-like extracellular serine protease inhibitors from P. infestans. Among these, EPI1 and EPI10 bind and inhibit the pathogenesis related (PR) P69B subtilisin-like serine protease of tomato. Here, we describe EPIC1 to EPIC4, a new family of P. infestans secreted proteins with similarity to cystatin-like protease inhibitor domains. Among these, the epiC1 and epiC2 genes lacked orthologs in Phytophthora sojae and Phytophthora ramorum, were relatively fast-evolving within P. infestans, and were up-regulated during infection of tomato, suggesting a role during P. infestans-host interactions. Biochemical functional analyses revealed that EPIC2B interacts with and inhibits a novel papain-like extracellular cysteine protease, termed Phytophthora Inhibited Protease 1 (PIP1). Characterization of PIP1 revealed that it is a PR protein closely related to Rcr3, a tomato apoplastic cysteine protease that functions in fungal resistance. Altogether, this and earlier studies suggest that interplay between host proteases of diverse catalytic families and pathogen inhibitors is a general defense-counterdefense process in plant-pathogen interactions. PMID- 17085510 TI - Physiological roles of the light, oxygen, or voltage domains of phototropin 1 and phototropin 2 in Arabidopsis. AB - Phototropins (phot1 and phot2) are plant blue-light receptors that mediate phototropism, chloroplast movement, stomatal opening, rapid inhibition of growth of etiolated seedlings, and leaf expansion in Arabidopsis (Arabidopsis thaliana). Their N-terminal region contains two light, oxygen, or voltage (LOV) domains, which bind flavin mononucleotide and form a covalent adduct between a conserved cysteine and the flavin mononucleotide chromophore upon photoexcitation. The C terminal region contains a serine/threonine kinase domain that catalyzes blue light-activated autophosphorylation. Here, we have transformed the phot1 phot2 (phot1-5 phot2-1) double mutant with PHOT expression constructs driven by the cauliflower mosaic virus 35S promoter. These constructs encode either wild-type phototropin or phototropin with one or both LOV-domain cysteines mutated to block their photochemistry. We selected multiple lines in each of the eight resulting categories of transformants for further physiological analyses. Specifically, we investigated whether LOV1 and LOV2 serve the same or different functions for phototropism and leaf expansion. Our results show that the LOV2 domain of phot1 plays a major role in phototropism and leaf expansion, as does the LOV2 domain of phot2. No complementation of phototropism or leaf expansion was observed for the LOV1 domain of phot1. However, phot2 LOV1 was unexpectedly found to complement phototropism to a considerable level. Similarly, transformants carrying a PHOT transgene with both LOV domains inactivated developed strong curvatures toward high fluence rate blue light. However, we found that the phot2-1 mutant is leaky and produces a small level of full-length phot2 protein. In vitro experiments indicate that cross phosphorylation can occur between functional phot2 and inactivated phot1 molecules. Such a mechanism may occur in vivo and therefore account for the functional activities observed in the PHOT transgenics with both lov domains inactivated. The implications of this mechanism with respect to phototropin function are discussed. PMID- 17085511 TI - Manganese deficiency in Chlamydomonas results in loss of photosystem II and MnSOD function, sensitivity to peroxides, and secondary phosphorus and iron deficiency. AB - For photoheterotrophic growth, a Chlamydomonas reinhardtii cell requires at least 1.7 x 10(7) manganese ions in the medium. At lower manganese ion concentrations (typically <0.5 microm), cells divide more slowly, accumulate less chlorophyll, and the culture reaches stationary phase at lower cell density. Below 0.1 microm supplemental manganese ion in the medium, the cells are photosynthetically defective. This is accompanied by decreased abundance of D1, which binds the Mn(4)Ca cluster, and release of the OEE proteins from the membrane. Assay of Mn superoxide dismutase (MnSOD) indicates loss of activity of two isozymes in proportion to the Mn deficiency. The expression of MSD3 through MSD5, encoding various isoforms of the MnSODs, is up-regulated severalfold in Mn-deficient cells, but neither expression nor activity of the plastid Fe-containing superoxide dismutase is changed, which contrasts with the dramatically increased MSD3 expression and plastid MnSOD activity in Fe-deficient cells. Mn-deficient cells are selectively sensitive to peroxide but not methyl viologen or Rose Bengal, and GPXs, APX, and MSRA2 genes (encoding glutathione peroxidase, ascorbate peroxidase, and methionine sulfoxide reductase 2) are slightly up regulated. Elemental analysis indicates that the Mn, Fe, and P contents of cells in the Mn-deficient cultures were reduced in proportion to the deficiency. A natural resistance-associated macrophage protein homolog and one of five metal tolerance proteins were induced in Mn-deficient cells but not in Fe-deficient cells, suggesting that the corresponding gene products may be components of a Mn(2+)-selective assimilation pathway. PMID- 17085512 TI - Reciprocal leaf and root expression of AtAmt1.1 and root architectural changes in response to nitrogen starvation. AB - Nitrogen is an essential macronutrient for plant growth and survival. Here, the temporal and spatial sensing of nitrogen starvation is analyzed in Arabidopsis (Arabidopsis thaliana). The promoter for the high-affinity ammonium transporter, AtAmt1.1, is shown to be a valid indicator for nitrogen status in leaves and roots. An AtAmt1.1-Gal4 transgene using three 5x upstream activating sequence driven reporters (luciferase, green fluorescent protein, and beta-glucuronidase) facilitated in vivo profiling at the whole-plant and cellular levels. The effects of nitrogen supply, light duration, light intensity, and carbon on the expression of the AtAmt1.1 gene in the roots and aerial tissues are reported. Under nitrogen starvation, high expression is observed in the roots and, under nitrogen sufficient conditions, high expression is observed in the leaves. This reciprocal regulation of AtAmt1.1 was confirmed by quantitative reverse transcription polymerase chain reaction, which was also used to quantitate expression of the five other Amt genes in Arabidopsis. Although some of these show tissue specificity (roots or leaves), none exhibit reciprocal regulation like the AtAmt1.1-encoded high-affinity transporter. This robust reciprocal expression suggests that Arabidopsis undergoes rapid resource reallocation in plants grown under different nitrogen supply regimens. Ultimately, nitrogen starvation mediated reallocation results in root architectural restructuring. We describe the precise timing and cellular aspects of this nitrogen limitation response. PMID- 17085513 TI - Knockout of the AtCESA2 gene affects microtubule orientation and causes abnormal cell expansion in Arabidopsis. AB - Complete cellulose synthesis is required to form functional cell walls and to facilitate proper cell expansion during plant growth. AtCESA2 is a member of the cellulose synthase A family in Arabidopsis (Arabidopsis thaliana) that participates in cell wall formation. By analysis of transgenic seedlings, we demonstrated that AtCESA2 was expressed in all organs, except root hairs. The atcesa2 mutant was devoid of AtCESA2 expression, leading to the stunted growth of hypocotyls in seedlings and greatly reduced seed production in mature plants. These observations were attributed to alterations in cell size as a result of reduced cellulose synthesis in the mutant. The orientation of microtubules was also altered in the atcesa2 mutant, which was clearly observed in hypocotyls and petioles. Complementary expression of AtCESA2 in atcesa2 could rescue the mutant phenotypes. Together, we conclude that disruption of cellulose synthesis results in altered orientation of microtubules and eventually leads to abnormal plant growth. We also demonstrated that the zinc finger-like domain of AtCESA2 could homodimerize, possibly contributing to rosette assemblies of cellulose synthase A within plasma membranes. PMID- 17085514 TI - Characterization of a divinyl ether biosynthetic pathway specifically associated with pathogenesis in tobacco. AB - In tobacco (Nicotiana tabacum), an elicitor- and pathogen-induced 9-lipoxygenase (LOX) gene, NtLOX1, is essential for full resistance to pathogens, notably to an incompatible race of Phytophthora parasitica var. nicotianae (Ppn race 0). In this work, we aimed to identify those oxylipins induced during attempted infection by Ppn race 0 and down-regulated in NtLOX1 antisense plants. Here we show that colneleic and colnelenic acids, which significantly inhibit germination of Ppn zoospores, are produced in roots of wild-type plants inoculated with Ppn, but are down-regulated in NtLOX1 antisense plants. A search for a tobacco gene encoding the enzyme involved in the formation of these divinyl ether (DVE) fatty acids resulted in the cloning and characterization of a DVE synthase (DES) clone (NtDES1). NtDES1 is a 9-DES, specifically converting fatty acid 9-hydroperoxides into DVE fatty acids. NtDES1 has the potential to act in combination with NtLOX1 because, in the presence of the two enzymes, linoleic and linolenic acids were converted in vitro into colneleic and colnelenic acids, respectively. In addition, the pattern of NtDES1 gene expression was quite similar to that of NtLOX1. Their transcripts were undetected in healthy tissues from different plant organs, and accumulated locally and transiently after elicitation and fungal infection, but not after wounding. Visualization of NtDES1-yellow fluorescent protein and NtLOX1-cyan fluorescent protein fusion proteins in tobacco leaves indicated that both localize in the cytosol and are excluded from plastids, consistent with the presumed location of the 9-LOX pathway in plants and the lack of transit peptides for NtLOX1 and NtDES1, respectively. Our data suggest that, in tobacco, NtDES1 and NtLOX1 act together and form DVEs in response to pathogen attack and that this class of oxylipins modulates in vivo the outcome of the tobacco-Ppn race 0 interaction. PMID- 17085515 TI - Variation of enzyme activities and metabolite levels in 24 Arabidopsis accessions growing in carbon-limited conditions. AB - Our understanding of the interaction of carbon (C) metabolism with nitrogen (N) metabolism and growth is based mainly on studies of responses to environmental treatments, and studies of mutants and transformants. Here, we investigate which metabolic parameters vary and which parameters change in a coordinated manner in 24 genetically diverse Arabidopsis (Arabidopsis thaliana) accessions, grown in C limited conditions. The accessions were grown in short days, moderate light, and high nitrate, and analyzed for rosette biomass, levels of structural components (protein, chlorophyll), total phenols and major metabolic intermediates (sugars, starch, nitrate, amino acids), and the activities of seven representative enzymes from central C and N metabolism. The largest variation was found for plant weight, reducing sugars, starch at the end of the night, and several enzyme activities. High levels of one sugar correlated with high levels of other sugars and starch, and a trend to increased amino acids, slightly lower nitrate, and higher protein. The activities of enzymes at the interface of C and N metabolism correlated with each other, but were unrelated to carbohydrates, amino acid levels, and total protein. Rosette weight was unrelated or showed a weak negative trend to sugar and amino acid contents at the end of the day in most of the accessions, and was negatively correlated with starch at the end of the night. Rosette weight was positively correlated with several enzyme activities. We propose that growth is not related to the absolute levels of starch, sugars, and amino acids; instead, it is related to flux, which is indicated by the enzymatic capacity to use these central resources. PMID- 17085516 TI - AIDS-related non-Hodgkin's lymphoma in the first decade of highly active antiretroviral therapy. AB - Highly active antiretroviral therapy (HAART) has had a dramatic effect on the natural history of HIV disease, reducing the incidence of opportunistic infections and Kaposi's sarcoma, and improving overall survival. Since HAART became available in 1996, the incidence of AIDS-related non-Hodgkin's lymphoma (NHL) has fallen, and although there has been no change in the clinical features at presentation, the overall survival of patients with AIDS-related NHL has improved. Prognosis is now determined chiefly by lymphoma-associated factors similar to those in the general population (the International Prognostic Index), although serum CD4 count at lymphoma diagnosis is an additional independent prognostic factor. The management of patients with AIDS-related NHL with either infusional chemotherapy or CHOP-like regimens (cyclophosphamide, doxorubicin, vincristine and prednisolone) achieves response and survival rates approaching those observed in the general population. However, careful attention should be paid to central nervous system chemoprophylaxis, opportunistic infection prophylaxis and potential drug interactions between cytotoxic and antiretroviral therapies. In the era of HAART, the goal of therapy for these patients is now complete remission rather than palliation. PMID- 17085517 TI - Lung inflammation and vascular remodeling after repeated allergen challenge detected noninvasively by MRI. AB - Magnetic resonance imaging (MRI) has been used previously to follow noninvasively inflammatory processes in rat acute models of lung inflammation. Here the technique was applied to a model involving repeated intratracheal administration of ovalbumin (OA). Anatomical MRI was performed at different time points with respect to a single or multiple OA challenges in Brown Norway rats actively sensitized to the allergen. Vascular permeability was assessed using dynamic contrast-enhanced MRI (DCE-MRI). Bronchoalveolar lavage (BAL) fluid analysis and histology were performed to validate the MRI data. The time course of MRI signals after a single OA challenge reached a maximum at 48 h and decreased significantly at 96 h. After the second and subsequent challenges, the maximum signal occurred at 6 h with a time-dependent decline over the remainder of the time course. A reduction of the inflammatory response following repeated administration of OA was also detected by BAL fluid analysis. The decrease in vascular permeability assessed by DCE-MRI in repeatedly OA-challenged rats was consistent with the thickening of the vascular wall for vessels of diameter up to 300 microm revealed by histology. Angiogenesis of vessels smaller than 30 microm was also detected histologically. These results suggest that MRI can be used to detect the inflammatory response and vascular remodeling associated with chronic airway inflammation in rat models involving repeated administration of allergen. As the contrast agent used in the DCE-MRI experiments is approved for clinical use, there is potential to translate the approach to patients. PMID- 17085518 TI - Cell-cell adhesion in lung endothelium. AB - Homotypic cell-cell adhesion is essential for tissue and organ development, remodeling, regeneration, and physiological function. Whereas a significant number of homotypic cell-cell adhesion molecules have been identified, much more is known about those concentrated in epithelia than in endothelia. Among the endothelial cell-cell adhesion molecules, very little is known that is specific to endothelium in the pulmonary and bronchial circulations. This review focuses primarily on homotypic cell-cell adhesion molecules that are or are likely to be important in lung endothelium. PMID- 17085519 TI - The evolutionary continuum from lung development to homeostasis and repair. AB - A functional, developmental, and comparative biological approach is probably the most effective way for arranging gene regulatory networks (GRNs) in their biological contexts. Evolutionary developmental biology allows comparison of GRNs during development across phyla. For lung evolution, the parathyroid hormone related protein (PTHrP) GRN exemplifies a continuum from ontogeny to phylogeny, homeostasis, and repair. PTHrP signaling between the lung endoderm and mesoderm stimulates lipofibroblast differentiation by downregulating the myofibroblast Wnt signaling pathway and upregulating the protein kinase A-dependent cAMP signaling pathway, inducing the lipofibroblast phenotype. Leptin secreted by the lipofibroblast, in turn, binds to its receptor on the alveolar type II cell, stimulating surfactant synthesis to ensure alveolar homeostasis. Failure of the PTHrP/PTHrP receptor signaling mechanism causes transdifferentiation of lipofibroblasts to myofibroblasts, which are the hallmark for lung fibrosis. We have shown that by targeting peroxisome proliferator-activated receptor gamma, the downstream target for lipofibroblast PTHrP signaling, we can prevent lung fibrosis. We speculate that the recapitulation of the myofibroblast phenotype during transdifferentiation is consistent with lung injury as lung evolution in reverse. Repair recapitulates ontogeny because it is programmed to express the cross talk between epithelium and mesoderm through evolution. This model demonstrates how epithelial-mesenchymal cross talk, when seen as a recapitulation of ontogeny and phylogeny (in both a forward and reverse direction), predicts novel, effective diagnostic and therapeutic targets. PMID- 17085520 TI - 17beta-Estradiol inhibits keratinocyte-derived chemokine production following trauma-hemorrhage. AB - Neutrophil infiltration is a key step in the development of organ dysfunction following trauma-hemorrhage (T-H). Although we have previously shown that 17beta estradiol (E2) prevents neutrophil infiltration and organ damage following T-H, the mechanism by which E2 inhibits neutrophil transmigration remains unknown. We hypothesized that E2 prevents neutrophil infiltration via modulation of keratinocyte-derived chemokine (KC), a major attractant for neutrophils. To examine this, male C3H/HeN mice were subjected to T-H or sham operation and thereafter resuscitated with Ringer lactate and E2 (1 mg/kg body wt) or vehicle. Animals were killed 2 h after resuscitation, and Kupffer cells were isolated. Plasma levels and Kupffer cell production capacities of KC, TNF-alpha, and IL-6 were determined by BD Cytometric Bead Arrays; lung mRNA expression of KC was measured with real-time PCR; myeloperoxidase activity assays were performed to determine neutrophil infiltration, and organ damage was assessed by edema formation. Treatment with E2 decreased systemic levels and restored Kupffer cell production of KC, TNF-alpha, and IL-6, as well as KC gene expression and protein in the lung. This was accompanied with a decrease in neutrophil infiltration and edema formation in the lung. These results suggest that E2 prevents lung neutrophil infiltration and organ damage in part by decreasing KC during posttraumatic immune response. PMID- 17085521 TI - Proproliferative phenotype of pulmonary microvascular endothelial cells. AB - Endothelial cells perform a number of important functions including release of vasodilators, control of the coagulation cascade, and restriction of solutes and fluid from the extravascular space. Regulation of fluid balance is of particular importance in the microcirculation of the lung where the loss of endothelial barrier function can lead to alveolar flooding and life-threatening hypoxemia. Significant heterogeneity exists between endothelial cells lining the microcirculation and cells from larger pulmonary arteries, however, and these differences may be relevant in restoring barrier function following vascular injury. Using well-defined populations of rat endothelial cells harvested from the pulmonary microcirculation [pulmonary microvascular endothelial cells (PMVEC)] and from larger pulmonary arteries [pulmonary artery endothelial cells (PAEC)], we compared their growth characteristics in low serum conditions. Withdrawal of serum inhibited proliferation and induced G0/G1 arrest in PAEC, whereas PMVEC failed to undergo G0/G1 arrest and continued to proliferate. Consistent with this observation, PMVEC had an increased cdk4 and cdk2 kinase activity with hyperphosphorylated (inactive) retinoblastoma (Rb) relative to PAEC as well as a threefold increase in cyclin D1 protein levels; overexpression of the cdk inhibitors p21Cip1/Waf1 and p27Kip1 induced G0/G1 arrest. While serum withdrawal failed to induce G0/G1 arrest in nonconfluent PMVEC, confluence was associated with hypophosphorylated Rb and growth arrest; loss of confluence led to resumption of growth. These data suggest that nonconfluent PMVEC continue to proliferate independently of growth factors. This proliferative characteristic may be important in restoring confluence (and barrier function) in the pulmonary microcirculation following endothelial injury. PMID- 17085522 TI - Anti-inflammatory effects of zinc and alterations in zinc transporter mRNA in mouse models of allergic inflammation. AB - There is clinical evidence linking asthma with the trace element, zinc (Zn). Using a mouse model of allergic inflammation, we have previously shown that labile Zn decreases in inflamed airway epithelium (Truong-Tran AQ, Ruffin RE, Foster PS, Koskinen AM, Coyle P, Philcox JC, Rofe AM, Zalewski PD. Am J Respir Cell Mol Biol 27: 286-296, 2002). Moreover, mild nutritional Zn deficiency worsens lung function. Recently, a number of proteins belonging to the Solute Carrier Family 39 (ZIP) and Solute Carrier Family 30 (ZnT) have been identified that bind Zn and regulate Zn homeostasis. Mice were sensitized, and subsequently aerochallenged, with ovalbumin to induce acute and chronic airway inflammation. Mice received 0, 54, or 100 microg of Zn intraperitoneally. Tissues were analyzed for Zn content and histopathology. Inflammatory cells were counted in bronchoalveolar lavage fluid. Cytokine and Zn transporter mRNA levels were determined by cDNA gene array and/or real-time PCR. Zn supplementation decreased bronchoalveolar lavage fluid eosinophils by 40 and 80%, and lymphocytes by 55 and 66%, in the acute and chronic models, respectively. Alterations in Zn transporter expression were observed during acute inflammation, including increases in ZIP1 and ZIP14 and decreases in ZIP4 and ZnT4. Zn supplementation normalized ZIP1 and ZIP14, but it did not affect mRNA levels of cytokines or their receptors. Our results indicate that inflammation-induced alterations in Zn transporter gene expression are directed toward increasing Zn uptake. Increases in Zn uptake may be needed to counteract the local loss of Zn in the airway and to meet an increased demand for Zn-dependent proteins. The reduction of inflammatory cells by Zn in the airways provides support for Zn supplementation trials in human asthmatic individuals. PMID- 17085523 TI - Altered cholesterol homeostasis in cultured and in vivo models of cystic fibrosis. AB - Determining how the regulation of cellular processes is impacted in cystic fibrosis (CF) is fundamental to understanding disease pathology and to identifying new therapeutic targets. In this study, unesterified cholesterol accumulation is observed in lung and trachea sections obtained from CF patients compared with non-CF tissues, suggesting an inherent flaw in cholesterol processing. An alternate staining method utilizing a fluorescent cholesterol probe also indicates improper lysosomal storage of cholesterol in CF cells. Excess cholesterol is also manifested by a significant increase in plasma membrane cholesterol content in both cultured CF cells and in nasal tissue excised from cftr(-/-) mice. Impaired intracellular cholesterol movement is predicted to stimulate cholesterol synthesis, a hypothesis supported by the observation of increased de novo cholesterol synthesis in lung and liver of cftr( /-) mice compared with controls. Furthermore, pharmacological inhibition of cholesterol transport is sufficient to cause CF-like elevation in cytokine production in wild-type cells in response to bacterial challenge but has no effect in CF cells. These data demonstrate via multiple methods in both cultured and in vivo models that cellular cholesterol homeostasis is inherently altered in CF. This perturbation of cholesterol homeostasis represents a potentially important process in CF pathogenesis. PMID- 17085524 TI - Cardiovascular and lung inflammatory effects induced by systemically administered diesel exhaust particles in rats. AB - Pollution by particulates has consistently been associated with increased cardiorespiratory morbidity and mortality. It has been suggested that ultrafine particles, of which diesel exhaust particles (DEP) are significant contributors, are able to translocate from the airways into the bloodstream in vivo. In the present study, we assessed the effect of systemic administration of DEP on cardiovascular and respiratory parameters. DEP were administered into the tail vein of rats, and heart rate, blood pressure, blood platelet activation, and lung inflammation were studied 24 h later. Doses of 0.02, 0.1, or 0.5 mg DEP/kg (8, 42, or 212 microg DEP/rat) induced a significant decrease of heart rate and blood pressure compared with saline-treated rats. Although the number of platelets was not affected, all the doses of DEP caused a shortening of the bleeding time. Similarly, in addition to triggering lung edema, the bronchoalveolar lavage analysis revealed the presence of neutrophil influx in DEP-treated rats in a dose dependent manner. We conclude that the presence of DEP in the systemic circulation leads not only to cardiovascular and haemostatic changes but it also triggers pulmonary inflammation. PMID- 17085525 TI - Role of Rho kinases in PKG-mediated relaxation of pulmonary arteries of fetal lambs exposed to chronic high altitude hypoxia. AB - An increase in Rho kinase (ROCK) activity is implicated in chronic hypoxia induced pulmonary hypertension. In the present study, we determined the role of ROCKs in cGMP-dependent protein kinase (PKG)-mediated pulmonary vasodilation of fetal lambs exposed to chronic hypoxia. Fourth generation pulmonary arteries were isolated from near-term fetuses ( approximately 140 days of gestation) delivered from ewes exposed to chronic high altitude hypoxia for approximately 110 days and from control ewes. In vessels constricted to endothelin-1, 8-bromoguanosine-cGMP (8-Br-cGMP) caused a smaller relaxation in chronically hypoxic (CH) vessels compared with controls. Rp-8-Br-PET-cGMPS, a PKG inhibitor, attenuated relaxation to 8-Br-cGMP in control vessels to a greater extent than in CH vessels. Y-27632, a ROCK inhibitor, significantly potentiated 8-Br-cGMP-induced relaxation of CH vessels and had only a minor effect in control vessels. The expression of PKG was increased but was not accompanied with an increase in the activity of the enzyme in CH vessels. The expression of type II ROCK and activity of ROCKs were increased in CH vessels. The phosphorylation of threonine (Thr)696 and Thr850 of the regulatory subunit MYPT1 of myosin light chain phosphatase was inhibited by 8 Br-cGMP to a lesser extent in CH vessels than in controls. The difference was eliminated by Y-27632. These results suggest that chronic hypoxia in utero attenuates PKG-mediated relaxation in pulmonary arteries, partly due to inhibition of PKG activity and partly due to enhanced ROCK activity. Increased ROCK activity may inhibit PKG action through increased phosphorylation of MYPT1 at Thr696 and Thr850. PMID- 17085526 TI - Downregulation of PCNA potentiates p21-mediated growth inhibition in response to hyperoxia. AB - Prolonged exposure to hyperoxia inhibits cell proliferation in G1 via increased expression of p21. While p21 inhibits proliferating cell nuclear antigen (PCNA) dependent DNA synthesis, it can also directly lower PCNA abundance; however, it is unclear whether loss of PCNA contributes to growth arrest. Here, we investigate how PCNA loss affects ability of p21 to exert G1 growth arrest of lung epithelial cells exposed to hyperoxia. In A549 cells that express p21 and growth arrest in G1 during hyperoxia, small interfering RNA (siRNA) knockdown of p21 led to G1 checkpoint bypass, increased cell death, and restoration of PCNA expression. Conditional overexpression of the PCNA binding domain of p21 in H1299 cells that do not normally express p21, or exposure to hyperoxia, caused a time dependent loss of PCNA. Titrating PCNA levels using siRNA to approximate the low amount observed in cells expressing p21 resulted in S phase arrest. While lowering PCNA by itself caused S phase arrest, the combination of hyperoxia and siRNA against PCNA dramatically reduced PCNA abundance resulting in G1 arrest. G1 growth arrest was markedly enhanced upon the addition of p21 to these cells. Our findings suggest a model in which reducing expression of the abundant protein PCNA allows the less abundant protein p21 to be more effective at suppressing the processivity functions of remaining PCNA, thereby fully exerting the G1 checkpoint. Given that high p21 expression is often associated with lower PCNA abundance, our findings are suggestive of a global growth inhibitory mechanism involving p21-mediated PCNA suppression. PMID- 17085527 TI - Thromboxane hypersensitivity in hypoxic pulmonary artery myocytes: altered TP receptor localization and kinetics. AB - Hypoxia-induced neonatal persistent pulmonary hypertension (PPHN) is characterized by sustained vasospasm and increased thromboxane (TxA2)-to prostacyclin ratio. We previously demonstrated that moderate hypoxia induces myocyte TxA2 hypersensitivity. Here, we examined TxA2 prostanoid receptor (TP-R) localization and kinetics following hypoxia to determine the mechanism of hypoxia induced TxA2 hypersensitivity. Primary cultured neonatal pulmonary artery myocytes were exposed to 10% O2 (hypoxic myocytes; HM) or 21% O2 (normoxic myocytes; NM) for 3 days. PPHN was induced in neonatal piglets by in vivo exposure to 10% FiO2 for 3 days. TP-R was studied in whole lung sections from pigs with hypoxic PPHN- and age-matched controls; intracellular localization was studied by immunocytochemistry. TP-R affinity was studied in cultured myocytes by saturation binding kinetics using 3H-SQ-29548 and competitive binding kinetics by coincubation with U-46619. Phosphorylation and coupling were examined in immunoprecipitated TP-R. We report distal propagation of TP-R expression in PPHN, extending to pulmonary arteries <50 microm. In HM, intracellular TP-R moves towards the perinuclear region, mirroring a change in endoplasmic reticulum (ER) morphology. TP-R kinetics also alter in HM membranes, with decreased Kd and Bmax (maximal binding sites). Additionally, in hypoxia, 3H-SQ-29548 is displaced at lower concentration of U-46619 than in normoxia, suggesting increased agonist affinity. Phosphorylation of serine residues on HM TP-R was significantly decreased compared with NM; this difference correlated with increased Galphaq coupling in hypoxia and was ablated by incubation with PKA. We conclude that the TP-R is normally desensitized in the neonatal pulmonary circuit by PKA-mediated regulatory phosphorylation, decreasing ligand affinity and coupling to Galphaq; this protection is lost following hypoxic exposure. Also, the appearance of TP-R in resistance arteries after development of hypoxic PPHN may contribute to increased pulmonary arterial pressure. PMID- 17085528 TI - Gender differences in management and outcome in non-ST-elevation acute coronary syndrome. AB - OBJECTIVE: To study gender differences in management and outcome in patients with non-ST-elevation acute coronary syndrome. DESIGN, SETTING AND PATIENTS: Cohort study of 53 781 consecutive patients (37% women) from the Register of Information and Knowledge about Swedish Heart Intensive care Admissions (RIKS-HIA), with a diagnosis of either unstable angina pectoris or non-ST-elevation myocardial infarction. All patients were admitted to intensive coronary care units in Sweden, between 1998 and 2002, and followed for 1 year. MAIN OUTCOME MEASURES: Treatment intensity and in-hospital, 30-day and 1-year mortality. RESULTS: Women were older (73 vs 69 years, p<0.001) and more likely to have a history of hypertension and diabetes, but less likely to have a history of myocardial infarction or revascularisation. After adjustment, there were no major differences in acute pharmacological treatment or prophylactic medication at discharge. Revascularisation was, however, even after adjustment, performed more often in men (OR 1.15; 95% CI, 1.09 to 1.21). After adjustment, there was no significant difference in in-hospital (OR 1.03; 95% CI, 0.94 to 1.13) or 30-days (OR 1.07; 95% CI, 0.99 to 1.15) mortality, but at 1 year being male was associated with higher mortality (OR 1.12; 95% CI, 1.06 to 1.19). CONCLUSION: Although women are somewhat less intensively treated, especially regarding invasive procedures, after adjustment for differences in background characteristics, they have better long-term outcomes than men. PMID- 17085529 TI - Pre-closure of femoral venous access sites used for large-sized sheath insertion with the Perclose device in adults undergoing cardiac intervention. AB - BACKGROUND: Interventional procedures in adults with congenital cardiac conditions often require insertion of large-sized sheaths into the femoral veins. Data on the use of suture-mediated devices for femoral venous access site closure are scant and no data are available regarding venous patency after device use. OBJECTIVE: To assess the efficacy of the 6Fr Perclose (Abbott Vascular Devices, CA, USA) suture-mediated device in achieving haemostasis and venous patency after closure. DESIGN AND SETTING: 146 consecutive patients (80 women, mean (SD) age 45 (14) years) undergoing closure of 205 femoral venous access sites in a tertiary cardiac centre were studied. All received heparin and were taking concomitant aspirin or clopidogrel, or both. The majority (98%) had a >or=10Fr sheath inserted. RESULTS: Immediate haemostasis was achieved in 202 (99%) sites. Two patients (1.4%) had a major complication. On follow-up (111 patients, mean (SD) 71 (33) days) there was no evidence of haematoma or fistula formation. Doppler studies from a subgroup of 43 (29%) patients (mean (SD) age 45 (15) years, mean (SD) follow-up 47 (18) days) showed a common femoral venous diameter of 11.6 (2.7) mm on the device closed right and 12.2 (2.5) mm on the left vein (p>0.05). All accessed veins were patent with no pseudoaneurysm or arteriovenous fistula formation. CONCLUSION: Pre-closure of large-size sheath femoral venous access sites using the suture-mediated Perclose device is efficacious in achieving rapid haemostasis in the presence of anticoagulation. Doppler follow-up shows no loss of venous patency or luminal venous diameter as compared with the contralateral side. PMID- 17085530 TI - Real-time three-dimensional dobutamine stress echocardiography for coronary artery disease diagnosis: validation with coronary angiography. AB - OBJECTIVE: To compare real-time three-dimensional echocardiography (RT3DE) with two-dimensional dobutamine stress echocardiography (2DE) for the detection of myocardial ischaemia, with angiographic validation of the results. METHODS: 56 patients (mean (SD) age 64.5 (6.2) years, 38 males), referred for coronary angiography, were examined by 2DE and RT3DE during the same dobutamine stress protocol. RESULTS: All 56 patients completed the stress protocol uneventfully. The mean (SD) acquisition time for the necessary views to evaluate all segments was 26.3 (2.5) s for RT3DE and 58.8 (3.7) s for 2DE (p<0.001). At peak stress, RT3DE had a higher wall-motion score index (1.25 (0.24) by 2DE, 1.30 (0.27) by RT3DE; p = 0.014). The regional wall-motion score for the four apical segments at peak stress was compared; it was 1.35 (0.55) by 2DE and 1.52 (0.69) by RT3DE (p = 0.003). The diagnostic parameters of 2DE versus RT3DE were: sensitivity 73% vs 78%, specificity 93% vs 89% and overall accuracy 86% vs 85%, respectively. In the left anterior descending artery territory, in particular, where RT3DE had higher regional wall-motion scores, it showed a tendency towards higher sensitivity (85% vs 78%), although this difference did not achieve statistical significance. CONCLUSION: RT3DE identifies wall-motion abnormalities more readily in the apical region than 2DE, which may explain the tendency towards higher sensitivity in the left anterior descending artery territory. RT3DE results were validated using angiography as reference and findings indicate diagnostic equivalence to 2DE, with the advantage of considerable shorter acquisition times. PMID- 17085531 TI - Fetal ECG: a novel predictor of atrioventricular block in anti-Ro positive pregnancies. AB - OBJECTIVE: Approximately 2.8% of pregnancies are Ro/La antibody positive. 3-15% of fetuses develop complete heart block (CHB). First-degree atrioventricular heart block (1 degrees AVB) is reported in a third of Ro/La fetuses but as most have a normal postnatal ECG this may reflect inadequacies of Doppler measurement techniques. METHODS: Comparison was made between mechanical (mPR) and electrical (ePR) intervals obtained prospectively using Doppler and non-invasive fetal ECG (fECG) in 52 consecutive Ro/La pregnancies in 46 women carrying 54 fetuses in an observational study at a fetal medicine unit. 121 mPR and 37 ePR intervals were recorded in 49 Ro/La fetuses. Five were referred with CHB and excluded. ePR was measured successfully in 35/37 (94%) and mPR was measured in all cases. 1 degrees AVB was defined as PR >95% CI. Logistic regression predicted abnormal final fetal rhythm from first mPR or ePR. RESULTS: The ePR model gave 66.7% sensitivity (6 of 8 final abnormal fetal rhythm cases were predicted correctly in fetuses >20 weeks) and 96.2% specificity. mPR gave 44.4% sensitivity (4 of 9 cases) and 88.5% specificity. Z scores for ePR (zPR) were calculated from 199 normal fetuses. The area under the receiver operator characteristic (ROC) curve was 0.88 (95% CI, 0.754 to 1.007). A cut-off of 1.65 gave a sensitivity of 87.5% and specificity of 95% for those with prolonged and normal ePR intervals, respectively. CONCLUSION: zPR is better than mPR at differentiating between normal and prolonged PR intervals, suggesting that fECG is the diagnostic tool of choice to investigate the natural history and therapy of conduction abnormalities in Ro/La pregnancies. PMID- 17085532 TI - Instantaneous effects of resynchronisation therapy on exercise performance in heart failure patients: the mechanistic role and predictive power of total isovolumic time. AB - BACKGROUND: Cardiac resynchronisation therapy improves peak oxygen uptake (peak VO(2)) 3-9 months after device implantation. In chronic heart failure, total isovolumic time (t-IVT) is a major determinant of peak VO(2) and of cardiac output at peak dobutamine stress. In selected patients, resynchronisation can instantaneously shorten t-IVT. We sought to determine the acute effect of resynchronisation on exercise performance and determine, with pharmacological stress echocardiography, the mechanism underlying this effect. METHODS AND RESULTS: Twenty-two patients with resynchronisation were studied within 3 months after device implantation. On a single study day, sequential cardiopulmonary exercise tests were performed during native activation (left bundle branch block) and resynchronisation (atrio-biventricular pacing) in random order. Total-IVT and cardiac output (at rest and peak dobutamine stress) were then measured in each activation mode. Resynchronisation acutely increased peak VO(2) by 1.6 (SD 1.5) ml/kg/min (p<0.001) and shortened peak stress t-IVT by 10 (SD 7) s/min (p<0.001), with the effects in individual patients showing a correlation (r = -0.46, p<0.05). Amongst all measurements during native activation, the best predictor of gain in peak VO(2) from resynchronisation was peak stress t-IVT (r = 0.71, p<0.001) with every increment of 5 s/min of peak stress t-IVT during native activation predicting an 8% gain in peak VO(2). No conventional measures during native activation at rest or on stress (including QRS duration, Tei index, tissue Doppler intraventricular delay, and resting t-IVT) added significant additional information. CONCLUSIONS: In eligible patients, resynchronisation can acutely augment peak VO(2), possibly through a mechanism of t-IVT shortening. Under native activation, long t-IVT during peak stress is the single best predictor of acute resynchronisation-mediated increment in peak VO(2). PMID- 17085533 TI - Prognosis and prognostic factors in patients with hypertrophic cardiomyopathy in Japan: results from a nationwide study. AB - OBJECTIVE: To investigate prognosis and prognostic factors in patients with hypertrophic cardiomyopathy (HCM) in Japan. DESIGN: A nationwide epidemiological study. SETTING: Hospitals selected randomly from among all hospitals in Japan. PATIENTS: Clinical and epidemiological information for 2155 patients with HCM were collected in 1999. MAIN OUTCOME MEASURES: Patients were classified on the basis of baseline prognostic factors. Survival rates up to 5 years were calculated by Cox's proportional hazard model for 1605 patients. RESULTS: During the follow-up period, 241 deaths were recorded. The crude 5-year survival rate for the entire cohort was 86% (95% CI 84 to 88), and annual mortality ranged from 2.2% to 3.0%. A higher cardiothoracic ratio on chest x ray (HR 1.61; 95% CI 1.26 to 2.05, with 1 SD (6.2%) increase), a lower left ventricular ejection fraction (HR 1.42; 95% CI 1.20 to 1.69, with 1 SD (13%) decrease) and the presence of left bundle branch block (HR 3.14; 95% CI 1.28 to 7.71) were independently associated with a poorer prognosis, whereas the presence of apical hypertrophy at baseline (HR 0.58; 95%CI 0.36 to 0.92) predicted a better chance of survival. CONCLUSIONS: The nationwide survey of patients with hypertrophic cardiomyopathy yielded important information on its prognosis and prognostic factors. These observations afford, for the first time, a measure of risk stratification in patients with HCM in Japan. PMID- 17085534 TI - Differential effects of lower body negative pressure and upright tilt on splanchnic blood volume. AB - Upright posture and lower body negative pressure (LBNP) both induce reductions in central blood volume. However, regional circulatory responses to postural changes and LBNP may differ. Therefore, we studied regional blood flow and blood volume changes in 10 healthy subjects undergoing graded lower-body negative pressure ( 10 to -50 mmHg) and 8 subjects undergoing incremental head-up tilt (HUT; 20 degrees , 40 degrees , and 70 degrees ) on separate days. We continuously measured blood pressure (BP), heart rate, and regional blood volumes and blood flows in the thoracic, splanchnic, pelvic, and leg segments by impedance plethysmography and calculated regional arterial resistances. Neither LBNP nor HUT altered systolic BP, whereas pulse pressure decreased significantly. Blood flow decreased in all segments, whereas peripheral resistances uniformly and significantly increased with both HUT and LBNP. Thoracic volume decreased while pelvic and leg volumes increased with HUT and LBNP. However, splanchnic volume changes were directionally opposite with stepwise decreases in splanchnic volume with LBNP and stepwise increases in splanchnic volume during HUT. Splanchnic emptying in LBNP models regional vascular changes during hemorrhage. Splanchnic filling may limit the ability of the splanchnic bed to respond to thoracic hypovolemia during upright posture. PMID- 17085535 TI - Cardiac-specific overexpression of insulin-like growth factor 1 attenuates aging associated cardiac diastolic contractile dysfunction and protein damage. AB - Aging is associated with hepatic growth hormone resistance resulting in a fall in serum insulin-like growth factor 1 (IGF-1) level. However, whether loss of IGF-1 contributes to cardiac aging is unclear. This study was designed to examine the effect of cardiac overexpression of IGF-1 on cardiomyocyte contractile function in young (3 mo) and old (26-28 mo) mice. Cardiomyocyte contractile function was evaluated, including peak shortening (PS), time to 90% PS, time to 90% relengthening (TR(90)), and maximal velocity of shortening/relengthening (+/ dL/dt). Levels of advanced glycation end product, protein carbonyl, sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA2a), phospholamban, and Na(+)/Ca(2+) exchanger were assessed by Western blot analysis. SERCA activity was measured by (45)Ca(2+) uptake. Aging induced a decline in plasma IGF-1 levels. Aged cells exhibited depressed +/-dL/dt, prolonged TR(90), and a steeper PS decline in response to increasing stimulus frequency compared with those in young myocytes. IGF-1 transgene alleviated aging-induced loss in plasma IGF-1 and aging induced mechanical defects with little effect in young mice. The beneficial effect of IGF-1 transgene on aging-associated cardiomyocyte contractile dysfunction was somewhat mimicked by short-term in vitro treatment of recombinant IGF-1 (500 nM). Advanced glycation end product and protein carbonyl levels were higher in aged mice, which were not affected by IGF-1. Expression of SERCA2a (but not Na(+)/Ca(2+) exchanger and phospholamban) and SERCA activity were reduced with aging, which was ablated by the IGF-1 transgene. Collectively, our data suggest a beneficial role of IGF-1 in aging-induced cardiac contractile dysfunction, possibly related to improved Ca(2+) uptake. PMID- 17085536 TI - Contribution of both Ca2+ entry and Ca2+ sensitization to the alpha1-adrenergic vasoconstriction of rat penile small arteries. AB - Sympathetic adrenergic nerves maintain the flaccid state of the penis through the tonic release of norepinephrine that contracts trabecular and arterial smooth muscle. Simultaneous measurements of intracellular Ca(2+) concentration ([Ca(2+)](i)) and tension and experiments with alpha-toxin-permeabilized arteries were performed in branches of the rat dorsal penile artery to investigate the intracellular Ca(2+) signaling pathways underlying alpha(1)-adrenergic vasoconstriction. Phenylephrine increased both [Ca(2+)](i) and tension, these increases being abolished by extracellular Ca(2+) removal and reduced by about 50% by the L-type Ca(2+) channel blocker nifedipine (0.3 microM). Non-L-type Ca(2+) entry through store-operated channels was studied by inhibiting the sarcoplasmic reticulum Ca(2+)-ATPase with cyclopiazonic acid (CPA). CPA (30 microM) induced variable phasic contractions that were abolished by extracellular Ca(2+) removal and by the store-operated channels antagonist 2 aminoethoxydiphenyl borate (2-APB, 50 microM) and largely inhibited by nifedipine (0.3 microM). CPA induced a sustained increase in [Ca(2+)](i) that was reduced in a Ca(2+)-free medium. Under conditions of L-type channels blockade, Ca(2+) readmission after store depletion with CPA evoked a sustained and marked elevation in [Ca(2+)](i) not coupled to contraction. 2-APB (50 microM) inhibited the rise in [Ca(2+)](i) evoked by CPA and the nifedipine-insensitive increases in both [Ca(2+)](i) and contraction elicited by phenylephrine. In alpha-toxin permeabilized penile arteries, activation of G proteins with guanosine 5'-O-(3 thiotriphosphate) and of the alpha(1)-adrenoceptor with phenylephrine both enhanced the myofilament sensitivity to Ca(2+). This Ca(2+) sensitization was reduced by selective inhibitors of PKC, tyrosine kinase (TK), and Rho kinase (RhoK) by 43%, 67%, and 82%, respectively. As a whole, the present data suggest the alpha(1)-adrenergic vasoconstriction in penile small arteries involves Ca(2+) entry through both L-type and 2-APB-sensitive receptor-operated channels, as well as Ca(2+) sensitization mechanisms mediated by PKC, TK, and RhoK. A capacitative Ca(2+) entry coupled to noncontractile functions of the smooth muscle cell is also demonstrated. PMID- 17085537 TI - Baroreceptor reflex regulation in anesthetized transgenic rats with low glia derived angiotensinogen. AB - Endogenous angiotensin (ANG) II and ANG-(1-7) act at the nucleus tractus solitarius (NTS) to differentially modulate neural control of the circulation. The role of these peptides endogenous to NTS on cardiovascular reflex function was investigated in transgenic rats with low brain angiotensinogen (Aogen) due to glial overexpression of an antisense to Aogen (ASrAOGEN) and in Sprague-Dawley (SD) rats. Arterial baroreceptor reflex sensitivity (BRS) for control of heart rate (HR) in response to increases in mean arterial pressure (MAP) was tested before and after bilateral microinjection of the angiotensin type 1 (AT(1)) receptor blocker candesartan or the ANG-(1-7) receptor blocker (d-Ala(7))-ANG-(1 7) into the NTS of urethane-chloralose-anesthetized ASrAOGEN and SD rats. Baseline MAP was higher in ASrAOGEN than in SD rats under anesthesia (P < 0.01). Injection of candesartan or (d-Ala(7))-ANG-(1-7) decreased MAP (P < 0.01) and HR (P < 0.05) in ASrAOGEN, but not SD, rats. The BRS at baseline was similar in ASrAOGEN and SD rats. Candesartan increased BRS by 41% in SD rats (P < 0.01) but was without effect in ASrAOGEN rats. In contrast, the reduction in BRS after (d Ala(7))-ANG-(1-7) administration was comparable in SD (31%) and ASrAOGEN rats (34%). These findings indicate that the absence of glia-derived Aogen is associated with 1) an increase in MAP under anesthesia mediated via AT(1) and ANG (1-7) receptors within the NTS, 2) the absence of an endogenous ANG II contribution to tonic inhibition of BRS, and 3) a continued contribution of endogenous ANG-(1-7) to tonic enhancement of BRS. PMID- 17085538 TI - Potentiation of the antihypertensive action of losartan by peripheral overexpression of the ANG II type 2 receptor. AB - Our previous studies demonstrated that peripheral overexpression of angiotensin II (ANG II) type 2 receptors (AT(2)R) prevents hypertension-induced cardiac hypertrophy and remodeling without altering high blood pressure. This, coupled with the observations that AT(2)R play a role in the antihypertensive actions of ANG II type 1 receptor (AT(1)R) blockers (ARBs), led us to propose that peripheral overexpression of AT(2)R would improve the antihypertensive action of losartan (Los) in Sprague-Dawley (SD) rats made hypertensive via chronic infusion of ANG II. Here we utilized adenoviral vector-mediated AT(2)R gene transfer to test this hypothesis. A single intracardiac injection of adenoviral vector containing genomic AT(2)R (G-AT(2)R) DNA and enhanced green fluorescent protein (EGFP) gene controlled by cytomegalovirus (CMV) promoters (Ad-G-AT(2)R-EGFP; 5 x 10(9) infectious units) into adult SD rats produced robust AT(2)R overexpression in cardiovascular tissues (kidney, lung, heart, aorta, mesenteric artery, and renal artery) that persisted for 3-5 days postinjection. By 7 days post viral injection, the overexpressed AT(2)R are reduced toward basal values in certain tissues (lung, kidney, and heart) and are undetectable in others (kidney and blood vessels). In two separate protocols, we demonstrated that the hypotensive effect of Los (0.125, 0.5, and 1.0 mg/kg iv) was significantly greater in the AT(2)R-overexpressing animals (-40.7 +/- 4.3, -41.8 +/- 4.8, and -48.1 +/- 2.6 mmHg, respectively) compared with control vector (Ad-CMV-EGFP)-treated rats ( 12.4 +/- 2.2, -20.2 +/- 3.4, and -27.3 +/- 3.4 mmHg, respectively). These results provide support for a depressor role of AT(2)R and the proposal that combined AT(2)R agonist and ARB treatment may be an improved therapeutic strategy for controlling hypertension. PMID- 17085539 TI - IL-1beta stimulates rat cardiac fibroblast migration via MAP kinase pathways. AB - The pro-inflammatory cytokines interleukin-1beta (IL-1beta), interleukin-6 (IL 6), and tumor necrosis factor-alpha (TNF-alpha) are elevated following acute myocardial infarction (MI) and have been implicated in the pathophysiology of cardiac disease progression. The cardiac fibroblast represents an important effector cell target for cytokine actions. In particular, cytokine-directed cardiac fibroblast migration is likely to impact both myocardial repair following acute MI and pathological myocardial remodeling in the progression to heart failure. In the present study, we examined the migratory response of neonatal rat cardiac fibroblasts to pro-inflammatory cytokines using modified Boyden chamber assays. On the basis of the knowledge of migration in other cell types, we hypothesized that members of the mitogen-activated protein kinase (MAPK) family may regulate this process. This possibility was addressed with the use of immunoblot detection of active phosphorylated MAPK species and pharmacological inhibitors for individual members of the MAPK cascades. IL-1beta stimulated robust and concentration-dependent increases in migration (maximum, 20-fold over control cells). TNF-alpha had lesser effect (fourfold increase over control). IL 6 did not induce migration. Activation of all three MAPK subfamilies (extracellular signal-regulated kinases, c-Jun NH(2)-terminal kinases, and p38) was shown to occur in response to cytokine stimulation. Fibroblast migration was attenuated by pharmacological inhibition of each MAPK subfamily. Understanding the regulation of cardiac fibroblast migration may provide insights in the search for therapies aimed at enhancing the functional nature of the remodeling process. PMID- 17085540 TI - Connexin 43 mediates endothelium-derived hyperpolarizing factor-induced vasodilatation in subcutaneous resistance arteries from healthy pregnant women. AB - The role of gap junctions in endothelium-derived hyperpolarizing factor (EDHF) mediated relaxation of human arteries was assessed using connexin mimetic peptides (CMPs) designated (37,43)Gap27, (40)Gap27, and (43)Gap26 according to homology with the major vascular connexins (Cx37, Cx40, and Cx43). Resistance arteries were obtained from subcutaneous fat biopsies of healthy pregnant women undergoing elective cesarean section. Endothelium-dependent vasodilatation to bradykinin (BK) was assessed using wire myography. N(omega)-nitro-l-arginine methyl ester (l-NAME) and indomethacin (nitric oxide synthase and cyclooxygenase inhibitors, respectively) attenuated maximal relaxation to BK (R(max)) by approximately 50%. Coincubation with l-NAME, indomethacin, and the combined CMPs ((37,43)Gap27, (40)Gap27, and (43)Gap26) almost abolished relaxation to BK (R(max) = 12.2 +/- 3.7%). In arteries incubated with l-NAME and indomethacin, the addition of either (37,43)Gap27 or (40)Gap27 had no significant effect on R(max), whereas (43)Gap26 caused marked inhibition (R(max) = 21 +/- 6.4%, P = 0.005 vs. l NAME plus indomethacin alone) that was similar to that of the triple combination. Endothelium-independent vasorelaxation was unaffected by CMPs, l-NAME, or indomethacin. Immunohistochemistry demonstrated Cx37, Cx40, and Cx43 expression in the endothelium and vascular smooth muscle. In pregnant women, EDHF-mediated vasorelaxation of subcutaneous resistance arteries is dependent on Cx43 and gap junctions. PMID- 17085541 TI - Insulin-induced activation of hypoxia-inducible factor-1 requires generation of reactive oxygen species by NADPH oxidase. AB - Hypoxia-inducible factor (HIF)-1 activation in response to hypoxia requires mitochondrial generation of reactive oxygen species (ROS). In contrast, the requirement of ROS for HIF-1 activation by growth factors like insulin remains unexplored. To explore that, insulin-sensitive hepatic cell HepG2 or cardiac muscle cell H9c2 cells were pretreated with NADPH oxidase inhibitor diphenyleneiodonium chloride (DPI) or apocynin and HIF-1 activation was tested by electrophoretic mobility shift and reporter gene assay. Antioxidants DPI or apocynin completely blocked insulin-stimulated HIF-1 activation. The restoration of HIF-1 activation by H(2)O(2) in DPI-pretreated cells not only confirmed the role of ROS but also identified H(2)O(2) as the responsible ROS. The role of NADPH oxidase was further confirmed by greater stimulation of HIF-1 during simultaneous treatment of suboptimal concentration of insulin along with NADPH but not by NADH. The role of oxidant generated by insulin is found to inhibit the protein tyrosine phosphatase as suggested by the following observations. First, tyrosine phosphatase-specific inhibitor sodium vanadate compensates DPI-inhibited HIF-1 activity. Second, sodium vanadate stimulates HIF-1 activation with suboptimal concentration of insulin. Third, DPI and pyrrolidene dithiocarbamate (PDTC) blocks insulin-receptor tyrosine kinase activation. The activity of phosphatidylinositol 3-kinase as evidenced by Akt phosphorylation, involved in HIF-1 activation, is also dependent on ROS generation by insulin. Finally, DPI pretreatment blocked insulin-stimulated expression of genes like VEGF, GLUT1, and ceruloplasmin. Overall, our data provide strong evidence for the essential role of NADPH oxidase-generated ROS in insulin-stimulated activation of HIF-1. PMID- 17085542 TI - Leg flow-mediated arterial dilation in elderly patients with heart failure and normal left ventricular ejection fraction. AB - BACKGROUND: flow-mediated arterial dilation (FMAD), an indicator of endothelial function, is reduced in patients with heart failure and reduced left ventricular ejection fraction (HFREF). Many elderly patients with heart failure exhibit a normal left ventricular ejection fraction (HFNEF). It is unknown whether FMAD is severely reduced in the elderly with HFNEF. METHODS AND RESULTS: 30 participants >60 yr of age, 11 healthy, 9 with HFNEF, and 10 with HFREF, underwent a cardiovascular magnetic resonance (CMR) assessment of FMAD in the superficial femoral artery followed within 48 h by symptom-limited exercise with expired gas analysis. Elderly patients with HFREF and HFNEF had severely reduced peak oxygen consumption (Vo(2 peak); 12 +/- 2 and 13 +/- 1 ml.kg(-1).min(-1), respectively) vs. their healthy age-matched contemporaries (20 +/- 3 ml.kg(-1).min(-1)). FMAD was 3.8 +/- 1.3% (0.85 +/- 0.22 mm(2)) in patients with HFREF; it was 12.1 +/- 3.6% (3.1 +/- 1.2 mm(2)) and 13.7 +/- 5.9% (3.9 +/- 1.7 mm(2)), respectively, in patients with HFNEF and age-matched healthy older individuals. After adjustment for age and gender, the association of FMAD with Vo(2) was high in healthy and HFREF subjects (P = 0.05 and 0.02, respectively) but less so in HFNEF participants (P = 0.58). CONCLUSIONS: elderly patients with HFNEF do not exhibit marked reduction in leg FMAD. These data suggest that mechanisms other than impaired femoral arterial endothelial function contribute to the severe exercise intolerance experienced by these individuals. PMID- 17085543 TI - Induction of cardioplegic arrest immediately activates the myocardial apoptosis signal pathway. AB - Myocardial ischemia-reperfusion, including cardioplegic arrest (CA), has been associated with cardiac apoptosis induction. However, the time course of apoptosis activation and the trigger mechanisms are still unclear. Because apoptosis inhibition may represent a novel therapeutic strategy for long-term myocardial preservation, we sought to investigate the time course of apoptosis signal-pathway induction during CA. As to method, Sprague-Dawley rats (300-350 g) were anesthetized, intubated, and mechanically ventilated. CA was initiated by infusion of ice-cold crystalloid solution (Custodiol, 10 ml/kg) into the aortic root, and hearts were rapidly excised and stored for 0, 30, 60, and 120 min in 0.9% sodium chloride solution (28 degrees C). In controls, no CA was initiated before removal and storage at 28 degrees C. In another group, calcium-rich cardioplegia was used, and an additional group received a caspase-8 inhibitor before CA induction. Left ventricular cytosolic extracts were isolated and investigated for the activity of caspase-3 and -6 (effector caspases) and caspase 8 and -9 (involved in extrinsic and intrinsic pathways of apoptosis induction). Fluorometric activity assays were performed by using specific substrates. As a result, activities of all tested caspases were significantly increased immediately after CA induction compared with controls. Administration of the caspase-8 inhibitor significantly reduced activities of all caspases. With calcium-rich cardioplegia, caspase activities were significantly lower compared with low-calcium CA. Control hearts also showed an increase of caspase activities during cold-storage ischemia without CA but had significantly different time courses compared with hearts with CA. In conclusion, our data show rapid apoptosis signal-pathway induction immediately following CA exposure. Thus apoptosis signal-pathway inhibition as a potential strategy for improved myocardial preservation would have the greatest effect when applied before CA exposure. PMID- 17085544 TI - The 5-hydroxytryptamine1A receptor agonist, (+)-8-hydroxy-2-(di-n-propylamino) tetralin, increases cardiac output and renal perfusion in rats subjected to hypovolemic shock. AB - The 5-hydroxytryptamine(1A) receptor agonist, (+)-8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT), raises blood pressure (BP) and venous tone in rats subjected to hemorrhagic shock. Here, BP, ascending aortic blood flow [i.e., estimate of cardiac output (CO)] and venous blood gases were measured to determine the hemodynamic effects of 8-OH-DPAT (30 nmol/kg i.v., n = 10), saline (n = 10), or an equipressor infusion of epinephrine (n = 10) in unanesthetized rats subjected to hemorrhagic shock (25 min of hypotensive hemorrhage, approximately 50 mm Hg). Renal and iliac blood flow were measured in separate groups of similarly hemorrhaged rats given the same dose of 8-OH-DPAT (n = 7) or saline (n = 6). Compared with saline treatment, 8-OH-DPAT produced a sustained rise in BP (+32 +/- 4 versus +9 +/- 2 mm Hg, 15 min after injection, P < 0.01) and CO (+27 +/- 5 versus +4 +/- 6 ml/min/kg, P < 0.01) but did not affect total peripheral resistance (TPR). Infusion of epinephrine reduced CO (-12 +/- 6 ml/min/kg, P < 0.01) and dramatically increased TPR [+0.37 +/- 0.11 versus +0.05 +/- 0.05 log (mm Hg/ml/min/kg), P < 0.01]. 8-OH-DPAT increased renal conductance (+7 +/- 1 versus +4 +/- 1 microl/min/mm Hg, P < 0.01) but did not significantly affect iliac conductance. 8-OH-DPAT attenuated further development of acidosis compared with either saline or epinephrine (-5.6 +/- 1.6 versus -13.0 +/- 2.0 versus -11.3 +/- 2.6 mmol/liter base excess 45 min after start of hemorrhage, both P < 0.01 versus 8-OH-DPAT). These data demonstrate that 8-OH-DPAT improves hemodynamics during circulatory shock, in part, through renal vasodilation and mobilizing of blood stores. PMID- 17085545 TI - Neutral endopeptidase up-regulation in isolated human umbilical artery: involvement in desensitization of bradykinin-induced vasoconstrictor effects. AB - Previous reports show that bradykinin B(2) receptors mediate contractile responses induced by bradykinin (BK) in human umbilical artery (HUA). However, although it has been reported that BK-induced responses can desensitize in several inflammatory models, the effects of prolonged in vitro incubation on BK induced vasoconstriction in HUA have not been studied. In isolated HUA rings, BK induced responses after a 5-h in vitro incubation showed a marked desensitization compared with responses at 2 h. Inhibition of either angiotensin-converting enzyme (ACE) or neutral endopeptidase (NEP), both BK-inactivating enzymes, failed to modify responses to BK at 2 h. After 5 h, ACE inhibition produced only a slight potentiation of BK-induced responses. In contrast, BK-induced vasoconstriction at 5 h was markedly potentiated by NEP inhibition. Moreover, NEP activity, measured by hydrolysis of its synthetic substrate (Z-Ala-Ala-Leu-p nitroanilide), showed a 2.4-fold increase in 5-h incubated versus 2-h incubated tissues, which was completely reversed by cycloheximide (CHX) treatment. Furthermore, CHX significantly potentiated BK-induced responses, suggesting that NEP-mediated kininase activity increase at 5 h depends on de novo protein synthesis. In addition, under NEP inhibition, CHX treatment failed to produce an additional potentiation of BK-induced vasoconstriction. Still, NEP up-regulation was confirmed by Western blot, showing a 2.1-fold increase in immunoreactive NEP in 5-h incubated versus 2-h incubated HUA. In summary, the present study provides strong pharmacological evidence that NEP is up-regulated and plays a key role in desensitization of BK-induced vasoconstriction after prolonged in vitro incubation in HUA. Our results provide new insights into the possible mechanisms involved in BK-induced response desensitization during sustained inflammatory conditions. PMID- 17085546 TI - Platelet-activating factor modulates activity of cyclic nucleotides in fetal ovine pulmonary vascular smooth muscle. AB - At birth, release of endogenous vasodilators such as nitric oxide and prostacyclin facilitate pulmonary vasodilation via the cyclic nucleotides, cGMP and cAMP. Interaction of cyclic nucleotides and platelet-activating factor (PAF) mediated responses in pulmonary vascular smooth muscle is not known. We studied the effects of cGMP and cAMP on PAF-mediated responses in ovine fetal intrapulmonary venous smooth muscle cells. Studies were done in hypoxia or normoxia with buffer with 8-Br-cGMP (BGMP) and 8-Br-cAMP (BAMP), as well as cGMP dependent protein kinase (PKG) and cAMP-dependent protein kinase (PKA) inhibitors. All groups were treated with 1 nM PAF and incubated for 30 min for the binding assay or 20 min for measurement of inositol 1,4,5-phosphate (IP(3)) production. BGMP and BAMP decreased PAF binding in normoxia by 63 and 14%, respectively. Incubations with the PKG inhibitor Rp-8-(4-chlorophenylthio) guanosine-3',5'-cyclic monophosphorothioate sodium and the PKA inhibitor Rp adenosine-3',5'-cyclic monophosphorothioate abrogated the inhibitory effects of BGMP and BAMP. PAF-stimulated IP(3) production was 8565 +/- 314 dpm/10(6) cells in hypoxia and 5418 +/- 118 dpm/10(6) cells in normoxia, a 40% decrease. BGMP attenuated PAF-stimulated IP(3) production by 67 and 37% in hypoxia and normoxia, respectively; the value for BAMP was 44% under both conditions. Pretreatment with PKG or PKA inhibitor abrogated BGMP and BAMP inhibition of IP(3) release. PAF receptor (PAFr) protein expression decreased in normoxia, but pretreatment with 10 nM PAF up-regulated PAFr expression. Pretreatment with PAF decreased expression and activities of PKG or PKA proteins in normoxia and hypoxia. Our data demonstrate the existence of cGMP/cAMP-PAF cross-talk in pulmonary vascular smooth muscle cells, which may be one mechanism by which PAFr-mediated vasoconstriction is down-regulated at birth. PMID- 17085547 TI - Escherichia coli HdeB is an acid stress chaperone. AB - We cloned, expressed, and purified the hdeB gene product, which belongs to the hdeAB acid stress operon. We extracted HdeB from bacteria by the osmotic-shock procedure and purified it to homogeneity by ion-exchange chromatography and hydroxyapatite chromatography. Its identity was confirmed by mass spectrometry analysis. HdeB has a molecular mass of 10 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis, which matches its expected molecular mass. We purified the acid stress chaperone HdeA in parallel in order to compare the two chaperones. The hdeA and hdeB mutants both display reduced viability upon acid stress, and only the HdeA/HdeB expression plasmid can restore their viability to close to the wild-type level, suggesting that both proteins are required for optimal protection of the bacterial periplasm against acid stress. Periplasmic extracts from both mutants aggregate at acidic pH, suggesting that HdeA and HdeB are required for protein solubilization. At pH 2, the aggregation of periplasmic extracts is prevented by the addition of HdeA, as previously reported, but is only slightly reduced by HdeB. At pH 3, however, HdeB is more efficient than HdeA in preventing periplasmic-protein aggregation. The solubilization of several model substrate proteins at acidic pH supports the hypothesis that, in vitro, HdeA plays a major role in protein solubilization at pH 2 and that both proteins are involved in protein solubilization at pH 3. Like HdeA, HdeB exposes hydrophobic surfaces at acidic pH, in accordance with the appearance of its chaperone properties at acidic pH. HdeB, like HdeA, dissociates from dimers at neutral pH into monomers at acidic pHs, but its dissociation is complete at pH 3 whereas that of HdeA is complete at a more acidic pH. Thus, we can conclude that Escherichia coli possesses two acid stress chaperones that prevent periplasmic protein aggregation at acidic pH. PMID- 17085548 TI - Membrane composition changes and physiological adaptation by Streptococcus mutans signal recognition particle pathway mutants. AB - Previously, we presented evidence that the oral cariogenic species Streptococcus mutans remains viable but physiologically impaired and sensitive to environmental stress when genes encoding the minimal conserved bacterial signal recognition particle (SRP) elements are inactivated. Two-dimensional gel electrophoresis of isolated membrane fractions from strain UA159 and three mutants (Deltaffh, DeltascRNA, and DeltaftsY) grown at pH 7.0 or pH 5.0 allowed us to obtain insight into the adaptation process and the identities of potential SRP substrates. Mutant membrane preparations contained increased amounts of the chaperones DnaK and GroES and ClpP protease but decreased amounts of transcription- and translation-related proteins, the beta subunit of ATPase, HPr, and several metabolic and glycolytic enzymes. Therefore, the acid sensitivity of SRP mutants might be caused in part by diminished ATPase activity, as well as the absence of an efficient mechanism for supplying ATP quickly at the site of proton elimination. Decreased amounts of LuxS were also observed in all mutant membranes. To further define physiological changes that occur upon disruption of the SRP pathway, we studied global gene expression in S. mutans UA159 (parent strain) and AH333 (Deltaffh mutant) using microarray analysis. Transcriptome analysis revealed up-regulation of 81 genes, including genes encoding chaperones, proteases, cell envelope biosynthetic enzymes, and DNA repair and replication enzymes, and down-regulation of 35 genes, including genes concerned with competence, ribosomal proteins, and enzymes involved in amino acid and protein biosynthesis. Quantitative real-time reverse transcription-PCR analysis of eight selected genes confirmed the microarray data. Consistent with a demonstrated defect in competence and the suggested impairment of LuxS-dependent quorum sensing, biofilm formation was significantly decreased in each SRP mutant. PMID- 17085549 TI - Global gene expression and phenotypic analysis of a Vibrio cholerae rpoH deletion mutant. AB - Vibrio cholerae, the cause of cholera, can grow in a variety of environments outside of human hosts. During infection, this pathogen must adapt to significant environmental alterations, including the elevated temperature of the human gastrointestinal tract. Sigma(32), an alternative sigma factor encoded by rpoH, activates transcription of genes involved in the heat shock response in several bacterial species. Here, we assessed the role of sigma(32) in V. cholerae physiology. In aggregate, our findings suggest that sigma(32) promotes V. cholerae growth at temperatures ranging at least from 15 degrees C to 42 degrees C. Growth of the rpoH mutant was severely attenuated within the suckling mouse intestine, suggesting that sigma(32)-regulated genes are critical for V. cholerae adaptation to conditions within the gastrointestinal tract. We defined the V. cholerae RpoH regulon by comparing the whole-genome transcription profiles of the wild-type and rpoH mutant strains after a temperature up-shift. Most of the V. cholerae genes expressed in an RpoH-dependent manner after heat shock encode proteins that influence protein fate, such as proteases and chaperones, or are of unknown function. Bioinformatic analyses of the microarray data were used to define a putative sigma(32) consensus binding sequence and subsequently to identify genes that are likely to be directly regulated by RpoH in the whole V. cholerae genome. PMID- 17085550 TI - Expression of the iron-activated nspA and secY genes in Neisseria meningitidis group B by Fur-dependent and -independent mechanisms. AB - Our whole-genome microarray studies of Neisseria meningitidis MC58 previously identified a set of 153 genes whose transcription was activated during growth in iron. In this study, Fur-mediated regulation of the iron-activated nspA gene was confirmed, whereas iron-activated regulation of the secY gene was demonstrated to be Fur independent. Analysis of the Fur binding sequences in the nspA gene and an additional iron-activated and Fur-regulated gene identified a hexameric (G/T)ATAAT unit in the operator regions of these genes similar to that observed in Fur- and iron-repressed genes. These studies indicate that the expression of the iron-activated nspA and secY genes in N. meningitidis occur by Fur-dependent and -independent mechanisms, respectively. PMID- 17085551 TI - Genomic structure and phylogeny of the plant pathogen Ralstonia solanacearum inferred from gene distribution analysis. AB - In the present study, we investigated the gene distribution among strains of the highly polymorphic plant pathogenic beta-proteobacterium Ralstonia solanacearum, paying particular attention to the status of known or candidate pathogenicity genes. Based on the use of comparative genomic hybridization on a pangenomic microarray for the GMI1000 reference strain, we have defined the conditions that allowed comparison of the repertoires of genes among a collection of 18 strains that are representative of the biodiversity of the R. solanacearum species. This identified a list of 2,690 core genes present in all tested strains. As a corollary, a list of 2,338 variable genes within the R. solanacearum species has been defined. The hierarchical clustering based on the distribution of variable genes is fully consistent with the phylotype classification that was previously defined from the nucleotide sequence analysis of four genes. The presence of numerous pathogenicity-related genes in the core genome indicates that R. solanacearum is an ancestral pathogen. The results establish the long coevolution of the two replicons that constitute the bacterial genome. We also demonstrate the clustering of variable genes in genomic islands. Most genomic islands are included in regions with an alternative codon usage, suggesting that they originate from acquisition of foreign genes through lateral gene transfers. Other genomic islands correspond to genes that have the same base composition as core genes, suggesting that they either might be ancestral genes lost by deletion in certain strains or might originate from horizontal gene transfers. PMID- 17085552 TI - How 34 pegs fit into 26 + 8 holes in the flagellar motor. PMID- 17085553 TI - A new look at bacteriophage lambda genetic networks. PMID- 17085554 TI - Regulation of gene expression in Streptococcus pneumoniae by response regulator 09 is strain dependent. AB - Recent murine studies have demonstrated that the role of response regulator 09 (RR09) of Streptococcus pneumoniae in virulence is different in different strains. In the present study, we used a murine pneumonia model of infection to assess the virulence of a TIGR4 rr09 mutant, and we found that TIGR4Deltarr09 was attenuated after intranasal infection. Furthermore, we investigated the in vitro transcriptional changes in pneumococcal rr09 mutants of two strains, D39 and TIGR4, by microarray analysis. The transcriptional profiles of the rr09 mutants of both strains had clear differences compared to the profiles of the parental wild-type strains. In D39Deltarr09, but not in TIGR4Deltarr09, genes involved in competence (e.g., comAB) were upregulated. In TIGR4, genes located on the rlrA pathogenicity islet, which are not present in the D39 genome, appeared to be regulated by RR09. Furthermore, several phosphotransferase systems (PTSs) believed to be involved in sugar uptake (e.g., the PTS encoded by sp0060 to sp0066) were strongly downregulated in D39Deltarr09, while they were not regulated by RR09 in TIGR4. To examine the role of one of these PTSs in virulence, D39Deltasp0063 was constructed and tested in a murine infection model. No difference between the virulence of this strain and the virulence of the wild type was found, indicating that downregulation of the sp0063 gene alone is not the cause of the avirulent phenotype of D39Deltarr09. Finally, expression of rr09 and expression of three of our identified RR09 targets during infection in mice were assessed. This in vivo experiment confirmed that there were differences between expression in wild-type strain TIGR4 and expression in the rr09 mutant, as well as differences between expression in wild-type strain D39 and expression in wild-type strain TIGR4. In conclusion, our results indicate that there is strain-specific regulation of pneumococcal gene expression by RR09. PMID- 17085555 TI - Complete and SOS-mediated response of Staphylococcus aureus to the antibiotic ciprofloxacin. AB - Staphylococcus aureus infections can be difficult to treat due to both multidrug resistance and the organism's remarkable ability to persist in the host. Persistence and the evolution of resistance may be related to several complex regulatory networks, such as the SOS response, which modifies transcription in response to environmental stress. To understand how S. aureus persists during antibiotic therapy and eventually emerges resistant, we characterized its global transcriptional response to ciprofloxacin. We found that ciprofloxacin induces prophage mobilization as well as significant alterations in metabolism, most notably the up-regulation of the tricarboxylic acid cycle. In addition, we found that ciprofloxacin induces the SOS response, which we show, by comparison of a wild-type strain and a non-SOS-inducible lexA mutant strain, includes the derepression of 16 genes. While the SOS response of S. aureus is much more limited than those of Escherichia coli and Bacillus subtilis, it is similar to that of Pseudomonas aeruginosa and includes RecA, LexA, several hypothetical proteins, and a likely error-prone Y family polymerase whose homologs in other bacteria are required for induced mutation. We also examined induced mutation and found that either the inability to derepress the SOS response or the lack of the LexA-regulated polymerase renders S. aureus unable to evolve antibiotic resistance in vitro in response to UV damage. The data suggest that up-regulation of the tricarboxylic acid cycle and induced mutation facilitate S. aureus persistence and evolution of resistance during antibiotic therapy. PMID- 17085556 TI - Substrate specificity of the Escherichia coli outer membrane protease OmpP. AB - Escherichia coli OmpP is an F episome-encoded outer membrane protease that exhibits 71% amino acid sequence identity with OmpT. These two enzymes cleave substrate polypeptides primarily between pairs of basic amino acids. We found that, like OmpT, purified OmpP is active only in the presence of lipopolysaccharide. With optimal peptide substrates, OmpP exhibits high catalytic efficiency (k(cat)/K(m) = 3.0 x 10(6) M(-1)s(-1)). Analysis of the extended amino acid specificity of OmpP by substrate phage revealed that both Arg and Lys are strongly preferred at the P1 and P1' sites of the enzyme. In addition, Thr, Arg, or Ala is preferred at P2; Leu, Ala, or Glu is preferred at P4; and Arg is preferred at P3'. Notable differences in OmpP and OmpT specificities include the greater ability of OmpP to accept Lys at the P1 or P1', site as well as the prominence of Ser at P3 in OmpP substrates. Likewise, the OmpP P1 site could better accommodate Ser; as a result, OmpP was able to cleave a peptide substrate between Ser-Arg about 120 times more efficiently than was OmpT. Interestingly, OmpP and OmpT cleave peptides with three consecutive Arg residues at different sites, a difference in specificity that might be important in the inactivation of cationic antimicrobial peptides. Accordingly, we show that the presence of an F' episome results in increased resistance to the antimicrobial peptide protamine both in ompT mutants and in wild-type E. coli cells. PMID- 17085557 TI - Extent of horizontal gene transfer in evolution of Streptococci of the salivarius group. AB - The phylogenetically closely related species Streptococcus salivarius and Streptococcus vestibularis are oral bacteria that are considered commensals, although they can also be found in human infections. The relationship between these two species and the relationship between strains isolated from carriers and strains responsible for invasive infections were investigated by multilocus sequence typing and additional sequence analysis. The clustering of several S. vestibularis alleles and the extent of genomic divergence at certain loci support the conclusion that S. salivarius and S. vestibularis are separate species. The level of sequence diversity in S. salivarius alleles is generally high, whereas that in S. vestibularis alleles is low at certain loci, indicating that the latter species might have evolved recently. Cluster analysis indicated that there has been genetic exchange between S. salivarius and S. vestibularis at three of the nine loci investigated. Horizontal gene transfer between streptococci belonging to the S. salivarius group and other oral streptococci was also detected at several loci. A high level of recombination in S. salivarius was revealed by allele index association and split decomposition sequence analyses. Commensal and infection-associated S. salivarius strains could not be distinguished by cluster analysis, suggesting that the pathogen isolates are opportunistic. Taken together, our results indicate that there is a high level of gene exchange that contributes to the evolution of two streptococcal species from the human oral cavity. PMID- 17085558 TI - Characterization of a higBA toxin-antitoxin locus in Vibrio cholerae. AB - Toxin-antitoxin (TA) loci, which were initially characterized as plasmid stabilization agents, have in recent years been detected on the chromosomes of numerous free-living bacteria. Vibrio cholerae, the causative agent of cholera, contains 13 putative TA loci, all of which are clustered within the superintegron on chromosome II. Here we report the characterization of the V. cholerae higBA locus, also known as VCA0391/2. Deletion of higA alone was not possible, consistent with predictions that it encodes an antitoxin, and biochemical analyses confirmed that HigA interacts with HigB. Transient exogenous expression of the toxin HigB dramatically slowed growth of V. cholerae and Escherichia coli and reduced the numbers of CFU by several orders of magnitude. HigB toxicity could be counteracted by simultaneous or delayed production of HigA, although HigA's effect diminished as the delay lengthened. Transcripts from endogenous higBA increased following treatment of V. cholerae with translational inhibitors, presumably due to reduced levels of HigA, which represses the higBA locus. However, no higBA-dependent cell death was observed in response to such stimuli. Thus, at least under the conditions tested, activation of endogenous HigB does not appear to be bactericidal. PMID- 17085559 TI - Population structure of plasmid-containing strains of Streptococcus mutans, a member of the human indigenous biota. AB - There are suggestions that the phylogeny of Streptococcus mutans, a member of the human indigenous biota that is transmitted mostly mother to child, might parallel the evolutionary history of its human host. The relatedness and phylogeny of plasmid-containing strains of S. mutans were examined based on chromosomal DNA fingerprints (CDF), a hypervariable region (HVR) of a 5.6-kb plasmid, the rRNA gene intergenic spacer region (IGSR), serotypes, and the genotypes of mutacin I and II. Plasmid-containing strains were studied because their genetic diversity was twice as great as that of plasmid-free strains. The CDF of S. mutans from unrelated human hosts were unique, except those from Caucasians, which were essentially identical. The evolutionary history of the IGSR, with or without the serotype and mutacin characters, clearly delineated an Asian clade. Also, a continuous association with mutacin II could be reconstructed through an evolutionary lineage with the IGSR, but not for serotype e. DNA sequences from the HVR of the plasmid produced a well-resolved phylogeny that differed from the chromosomal phylogeny, indicating that the horizontal transfer of the plasmid may have occurred multiple times. The plasmid phylogeny was more congruent with serotype e than with mutacin II evolution, suggesting a possible functional correlation. Thus, the history of this three-tiered relationship between human, bacterium, and plasmid supported both coevolution and independent evolution. PMID- 17085560 TI - Proteolytic adaptor for transfer-messenger RNA-tagged proteins from alpha proteobacteria. AB - We have identified an analog of SspB, the proteolytic adaptor for transfer messenger RNA (tmRNA)-tagged proteins, in Caulobacter crescentus. C. crescentus SspB shares limited sequence similarity with Escherichia coli SspB but binds the tmRNA tag in vitro and is required for optimal proteolysis of tagged proteins in vivo. PMID- 17085561 TI - CC1, a novel crenarchaeal DNA binding protein. AB - The genomes of the related crenarchaea Pyrobaculum aerophilum and Thermoproteus tenax lack any obvious gene encoding a single-stranded DNA binding protein (SSB). SSBs are essential for DNA replication, recombination, and repair and are found in all other genomes across the three domains of life. These two archaeal genomes also have only one identifiable gene encoding a chromatin protein (the Alba protein), while most other archaea have at least two different abundant chromatin proteins. We performed a biochemical screen for novel nucleic acid binding proteins present in cell extracts of T. tenax. An assay for proteins capable of binding to a single-stranded DNA oligonucleotide resulted in identification of three proteins. The first protein, Alba, has been shown previously to bind single stranded DNA as well as duplex DNA. The two other proteins, which we designated CC1 (for crenarchaeal chromatin protein 1), are very closely related to one another, and homologs are restricted to the P. aerophilum and Aeropyrum pernix genomes. CC1 is a 6-kDa, monomeric, basic protein that is expressed at a high level in T. tenax. This protein binds single- and double-stranded DNAs with similar affinities. These properties are consistent with a role for CC1 as a crenarchaeal chromatin protein. PMID- 17085562 TI - Evolutionary genomics of lactic acid bacteria. PMID- 17085563 TI - Minimum length requirement of the flexible N-terminal translocation subdomain of colicin E3. AB - The 315-residue N-terminal T domain of colicin E3 functions in translocation of the colicin across the outer membrane through its interaction with outer membrane proteins including the OmpF porin. The first 83 residues of the T domain are known from structure studies to be disordered. This flexible translocation subdomain contains the TolB box (residues 34 to 46) that must cross the outer membrane in an early translocation event, allowing the colicin to bind to the TolB protein in the periplasm. In the present study, it was found that cytotoxicity of the colicin requires a minimum length of 19 to 23 residues between the C terminus (residue 46) of the TolB box and the end of the flexible subdomain (residue 83). Colicin E3 molecules of sufficient length display normal binding to TolB and occlusion of OmpF channels in vitro. The length of the N terminal subdomain is critical because it allows the TolB box to cross the outer membrane and interact with TolB. It is proposed that the length constraint is a consequence of ordered structure in the downstream segment of the T domain (residues 84 to 315) that prevents its insertion through the outer membrane via a translocation pore that includes OmpF. PMID- 17085564 TI - Heme concentration dependence and metalloporphyrin inhibition of the system I and II cytochrome c assembly pathways. AB - Studies have indicated that specific heme delivery to apocytochrome c is a critical feature of the cytochrome c biogenesis pathways called system I and II. To determine directly the heme requirements of each system, including whether other metal porphyrins can be incorporated into cytochromes c, we engineered Escherichia coli so that the natural system I (ccmABCDEFGH) was deleted and exogenous porphyrins were the sole source of porphyrins (Delta hemA). The engineered E. coli strains that produced recombinant system I (from E. coli) or system II (from Helicobacter) facilitated studies of the heme concentration dependence of each system. Using this exogenous porphyrin approach, it was shown that in system I the levels of heme used are at least fivefold lower than the levels used in system II, providing an important advantage for system I. Neither system could assemble holocytochromes c with other metal porphyrins, suggesting that the attachment mechanism is specific for Fe protoporphyrin. Surprisingly, Zn and Sn protoporphyrins are potent inhibitors of the pathways, and exogenous heme competes with this inhibition. We propose that the targets are the heme binding proteins in the pathways (CcmC, CcmE, and CcmF for system I and CcsA for system II). PMID- 17085565 TI - Influence of wall teichoic acid on lysozyme resistance in Staphylococcus aureus. AB - Staphylococcus aureus peptidoglycan (PG) is completely resistant to the hydrolytic activity of lysozyme. Here we show that modifications in PG by O acetylation, wall teichoic acid, and a high degree of cross-linking contribute to this resistance. PMID- 17085566 TI - Complete and integrated pyrene degradation pathway in Mycobacterium vanbaalenii PYR-1 based on systems biology. AB - Mycobacterium vanbaalenii PYR-1 was the first bacterium isolated by virtue of its ability to metabolize the high-molecular-weight polycyclic aromatic hydrocarbon (PAH) pyrene. We used metabolic, genomic, and proteomic approaches in this investigation to construct a complete and integrated pyrene degradation pathway for M. vanbaalenii PYR-1. Genome sequence analyses identified genes involved in the pyrene degradation pathway that we have proposed for this bacterium. To identify proteins involved in the degradation, we conducted a proteome analysis of cells exposed to pyrene using one-dimensional gel electrophoresis in combination with liquid chromatography-tandem mass spectrometry. Database searching performed with the M. vanbaalenii PYR-1 genome resulted in identification of 1,028 proteins with a protein false discovery rate of <1%. Based on both genomic and proteomic data, we identified 27 enzymes necessary for constructing a complete pathway for pyrene degradation. Our analyses indicate that this bacterium degrades pyrene to central intermediates through o-phthalate and the beta-ketoadipate pathway. Proteomic analysis also revealed that 18 enzymes in the pathway were upregulated more than twofold, as indicated by peptide counting when the organism was grown with pyrene; three copies of the terminal subunits of ring-hydroxylating oxygenase (NidAB2, MvanDraft_0817/0818, and PhtAaAb), dihydrodiol dehydrogenase (MvanDraft_0815), and ring cleavage dioxygenase (MvanDraft_3242) were detected only in pyrene-grown cells. The results presented here provide a comprehensive picture of pyrene metabolism in M. vanbaalenii PYR-1 and a useful framework for understanding cellular processes involved in PAH degradation. PMID- 17085567 TI - Transcriptional regulation of the yghJ-pppA-yghG-gspCDEFGHIJKLM cluster, encoding the type II secretion pathway in enterotoxigenic Escherichia coli. AB - The gene cluster gspCDEFGHIJKLM codes for various structural components of the type II secretion pathway which is responsible for the secretion of heat-labile enterotoxin by enterotoxigenic Escherichia coli (ETEC). In this work, we used a variety of molecular approaches to elucidate the transcriptional organization of the ETEC type II secretion system and to unravel the mechanisms by which the expression of these genes is controlled. We showed that the gspCDEFGHIJKLM cluster and three other upstream genes, yghJ, pppA, and yghG, are cotranscribed and that a promoter located in the upstream region of yghJ plays a major role in the expression of this 14-gene transcriptional unit. Transcription of the yghJ promoter was repressed 168-fold upon a temperature downshift from 37 degrees C to 22 degrees C. This temperature-induced repression was mediated by the global regulatory proteins H-NS and StpA. Deletion mutagenesis showed that the promoter region encompassing positions -321 to +301 relative to the start site of transcription of yghJ was required for full repression. The yghJ promoter region is predicted to be highly curved and bound H-NS or StpA directly. The binding of H-NS or StpA blocked transcription initiation by inhibiting promoter open complex formation. Unraveling the mechanisms of regulation of type II secretion by ETEC enhances our understanding of the pathogenesis of ETEC and other pathogenic varieties of E. coli. PMID- 17085568 TI - From oil to bioplastics, a dream come true? PMID- 17085569 TI - Naturally occurring adenines within mRNA coding sequences affect ribosome binding and expression in Escherichia coli. AB - Translation initiation requires the precise positioning of a ribosome at the start codon. The major signals of bacterial mRNA that direct the ribosome to a translational start site are the Shine-Dalgarno (SD) sequence within the untranslated leader and the start codon. Evidence for the presence of many non-SD led genes in prokaryotes provides a motive for studying additional interactions between ribosomes and mRNA that contribute to translation initiation. A high incidence of adenines has been reported downstream of the start codon for many Escherichia coli genes, and addition of downstream adenine-rich sequences increases expression from several genes in E. coli. Here we describe site directed mutagenesis of the E. coli aroL, pncB, and cysJ coding sequences that was used to assess the contribution of naturally occurring adenines to in vivo expression and in vitro ribosome binding from mRNAs with different SD-containing untranslated leaders. Base substitutions that decreased the downstream adenines by one or two nucleotides decreased expression significantly from aroL-, pncB-, and cysJ-lacZ fusions; mutations that increased downstream adenines by one or two nucleotides increased expression significantly from aroL- and cysJ-lacZ fusions. Using primer extension inhibition (toeprint) and filter binding assays to measure ribosome binding, the changes in in vivo expression correlated closely with changes in in vitro ribosome binding strength. Our data are consistent with a model in which downstream adenines influence expression through their effects on the mRNA-ribosome association rate and the amount of ternary complex formed. This work provides evidence that adenine-rich sequence motifs might serve as a general enhancer of E. coli translation. PMID- 17085570 TI - Bacillus subtilis gene cluster involved in calcium carbonate biomineralization. AB - Calcium carbonate precipitation, a widespread phenomenon among bacteria, has been investigated due to its wide range of scientific and technological implications. Nevertheless, little is known of the molecular mechanisms by which bacteria foster calcium carbonate mineralization. In our laboratory, we are studying calcite formation by Bacillus subtilis, in order to identify genes involved in the biomineralization process. A previous screening of UV mutants and of more than one thousand mutants obtained from the European B. subtilis Functional Analysis project allowed us to isolate strains altered in the precipitation phenotype. Starting from these results, we focused our attention on a cluster of five genes (lcfA, ysiA, ysiB, etfB, and etfA) called the lcfA operon. By insertional mutagenesis, mutant strains carrying each of the five genes were produced. All of them, with the exception of the strain carrying the mutated lcfA operon, were unable to form calcite crystals. By placing transcription under IPTG (isopropyl-beta-d-thiogalactopyranoside) control, the last gene, etfA, was identified as essential for the precipitation process. To verify cotranscription in the lcfA operon, reverse transcription-PCR experiments were performed and overlapping retrocotranscripts were found comprising three adjacent genes. The genes have putative functions linked to fatty acid metabolism. A link between calcium precipitation and fatty acid metabolism is suggested. PMID- 17085571 TI - Cys303 in the histidine kinase PhoR is crucial for the phosphotransfer reaction in the PhoPR two-component system in Bacillus subtilis. AB - The PhoPR two-component system activates or represses Pho regulon genes to overcome a phosphate deficiency. The Pho signal transduction network is comprised of three two-component systems, PhoPR, ResDE, and Spo0A. Activated PhoP is required for expression of ResDE from the resA promoter, while ResD is essential for 80% of Pho induction, establishing a positive feedback loop between these two component systems to amplify the signal received by the Pho system. The role of ResD in the Pho response is via production of terminal oxidases. Reduced quinones inhibit PhoR autophosphorylation in vitro, and it was proposed that the expression of terminal oxidases leads to oxidation of the quinone pool, thereby relieving the inhibition. We show here that the reducing environment generated by dithiothreitol (DTT) in vivo inhibited Pho induction in a PhoR-dependent manner, which is in agreement with our previous in vitro data. A strain containing a PhoR variant, PhoR(C303A), exhibited reduced Pho induction and remained sensitive to inhibition by DTT, suggesting that the mechanisms for Pho reduction via PhoR(C303A) and DTT are different. PhoR and PhoR(C303A) were similar with regard to cellular concentration, limited proteolysis patterns, rate of autophosphorylation, stability of PhoR approximately P, and inhibition of autophosphorylation by DTT. Phosphotransfer between PhoR approximately P or PhoR(C303A) approximately P and PhoP occurred rapidly; most label from PhoR approximately P was transferred to PhoP, but only 10% of the label from PhoR(C303A) approximately P was associated with PhoP, while 90% was released as inorganic phosphate. No difference in PhoP approximately P or PhoR autophosphatase activity was observed between PhoR and PhoR(C303A) that would explain the release of inorganic phosphate. Our data are consistent with a role for PhoR(C303) in PhoR activity via stabilization of the phosphoryl-protein intermediate(s) during phosphotransfer from PhoR approximately P to PhoP, which is stabilization that is required for efficient production of PhoP approximately P. PMID- 17085572 TI - Interference of components of the phosphoenolpyruvate phosphotransferase system with the central virulence gene regulator PrfA of Listeria monocytogenes. AB - Analysis of Listeria monocytogenes ptsH, hprK, and ccpA mutants defective in carbon catabolite repression (CCR) control revealed significant alterations in the expression of PrfA-dependent genes. The hprK mutant showed high up-regulation of PrfA-dependent virulence genes upon growth in glucose-containing medium whereas expression of these genes was even slightly down-regulated in the ccpA mutant compared to the wild-type strain. The ptsH mutant could only grow in a rich culture medium, and here the PrfA-dependent genes were up-regulated as in the hprK mutant. As expected, HPr-Ser-P was not produced in the hprK and ptsH mutants and synthesized at a similar level in the ccpA mutant as in the wild-type strain. However, no direct correlation was found between the level of HPr-Ser-P or HPr-His-P and PrfA activity when L. monocytogenes was grown in minimal medium with different phosphotransferase system (PTS) carbohydrates. Comparison of the transcript profiles of the hprK and ccpA mutants with that of the wild-type strain indicates that the up-regulation of the PrfA-dependent virulence genes in the hprK mutant correlates with the down-regulation of genes known to be controlled by the efficiency of PTS-mediated glucose transport. Furthermore, growth in the presence of the non-PTS substrate glycerol results in high PrfA activity. These data suggest that it is not the component(s) of the CCR or the common PTS pathway but, rather, the component(s) of subsequent steps that seem to be involved in the modulation of PrfA activity. PMID- 17085573 TI - Mutational analysis of the flagellar protein FliG: sites of interaction with FliM and implications for organization of the switch complex. AB - The switch complex at the base of the bacterial flagellum is essential for flagellar assembly, rotation, and switching. In Escherichia coli and Salmonella, the complex contains about 26 copies of FliG, 34 copies of FliM, and more then 100 copies of FliN, together forming the basal body C ring. FliG is involved most directly in motor rotation and is located in the upper (membrane-proximal) part of the C ring. A crystal structure of the middle and C-terminal parts of FliG shows two globular domains connected by an alpha-helix and a short extended segment. The middle domain of FliG has a conserved surface patch formed by the residues EHPQ(125-128) and R(160) (the EHPQR motif), and the C-terminal domain has a conserved surface hydrophobic patch. To examine the functional importance of these and other surface features of FliG, we made mutations in residues distributed over the protein surface and measured the effects on flagellar assembly and function. Mutations preventing flagellar assembly occurred mainly in the vicinity of the EHPQR motif and the hydrophobic patch. Mutations causing aberrant clockwise or counterclockwise motor bias occurred in these same regions and in the waist between the upper and lower parts of the C-terminal domain. Pull down assays with glutathione S-transferase-FliM showed that FliG interacts with FliM through both the EHPQR motif and the hydrophobic patch. We propose a model for the organization of FliG and FliM subunits that accounts for the FliG-FliM interactions identified here and for the different copy numbers of FliG and FliM in the flagellum. PMID- 17085574 TI - Functional analysis of the carboxy-terminal region of Bacillus subtilis TnrA, a MerR family protein. AB - The Bacillus subtilis TnrA transcription factor belongs to the MerR family of proteins and regulates gene expression during nitrogen-limited growth. When B. subtilis cells are grown with excess nitrogen, feedback-inhibited glutamine synthetase forms a protein-protein complex with TnrA that prevents TnrA from binding to DNA. The C-terminal region of TnrA is required for the interaction with glutamine synthetase. Alanine scanning mutagenesis of the C-terminal region of TnrA identified three classes of mutants that altered the regulation by glutamine synthetase. While expression of the TnrA-regulated amtB gene was expressed constitutively in the class I (M96A, Q100A, and A103G) and class II (L97A, L101A, and F105A) mutants, the class II mutants were unable to grow on minimal medium unless a complex mixture of amino acids was present. The class III tnrA mutants (R93A, G99A, N102A, H104A, and Y107A mutants) were partially defective in the regulation of TnrA activity. In vitro experiments showed that feedback-inhibited glutamine synthetase had a significantly reduced ability to inhibit the DNA-binding activity of several class I and class II mutant TnrA proteins. A coiled-coil homology model of the C-terminal region of TnrA is used to explain the properties of the class I and II mutant proteins. The C-terminal region of TnrA corresponds to a dimerization domain in other MerR family proteins. Surprisingly, gel filtration and cross-linking analysis showed that a truncated TnrA protein which contained only the N-terminal DNA binding domain was dimeric. The implications of these results for the structure of TnrA are discussed. PMID- 17085575 TI - Glycosylation of Pseudomonas aeruginosa strain Pa5196 type IV pilins with mycobacterium-like alpha-1,5-linked d-Araf oligosaccharides. AB - Pseudomonas aeruginosa is a gram-negative bacterium that uses polar type IV pili for adherence to various materials and for rapid colonization of surfaces via twitching motility. Within the P. aeruginosa species, five distinct alleles encoding variants of the structural subunit PilA varying in amino acid sequence, length, and presence of posttranslational modifications have been identified. In this work, a combination of mass spectrometry and nuclear magnetic resonance spectroscopy was used to identify a novel glycan modification on the pilins of the group IV strain Pa5196. Group IV pilins continued to be modified in a lipopolysaccharide (wbpM) mutant of Pa5196, showing that, unlike group I strains, the pilins of group IV are not modified with the O-antigen unit of the background strain. Instead, the pilin glycan was determined to be an unusual homo-oligomer of alpha-1,5-linked d-arabinofuranose (d-Araf). This sugar is uncommon in prokaryotes, occurring mainly in the cell wall arabinogalactan and lipoarabinomannan (LAM) polymers of mycobacteria, including Mycobacterium tuberculosis and Mycobacterium leprae. Antibodies raised against M. tuberculosis LAM specifically identified the glycosylated pilins from Pa5196, confirming that the glycan is antigenically, as well as chemically, identical to those of Mycobacterium. P. aeruginosa Pa5196, a rapidly growing strain of low virulence that expresses large amounts of glycosylated type IV pilins on its surface, represents a genetically tractable model system for elucidation of alternate pathways for biosynthesis of d-Araf and its polymerization into mycobacterium like alpha-1,5-linked oligosaccharides. PMID- 17085576 TI - Characterization of Helicobacter pylori lytic transglycosylases Slt and MltD. AB - Peptidoglycan (PG) is a cell wall heteropolymer that is essential for cell integrity. PG hydrolases participate in correct assembly of the PG layer and have been shown to be required for cell division, cell daughter separation, and maintenance of bacterial morphology. In silico analysis of the Helicobacter pylori genome resulted in identification of three potential hydrolases, Slt, MltD, and AmiA. This study was aimed at determining the roles of the putative lytic transglycosylases, Slt and MltD, in H. pylori morphology, growth, and PG metabolism. Strain 26695 single mutants were constructed using a nonpolar kanamycin cassette. The slt and mltD mutants formed normal bacillary and coccoid bacteria in the exponential and stationary phases, respectively. The slt and mltD mutants had growth rates comparable to the growth rate of the parental strain. However, the mltD mutant exhibited enhanced survival in the stationary phase compared to the wild type or the slt mutant. PG was purified from exponentially growing bacteria and from bacteria in the stationary phase, and its muropeptide composition was analyzed by high-pressure liquid chromatography. This analysis revealed changes in the muropeptide composition indicating that MltD and Slt have lytic transglycosylase activities. Glycan strand analysis suggested that Slt and MltD have exo and endo types of lytic transglycosylase activity, indicating that Slt is involved mainly in PG turnover and MltD is involved mainly in rearrangement of the PG layer. In this study, we determined the distinct roles of the lytic transglycosylases Slt and MltD in PG metabolism. PMID- 17085577 TI - The C-terminal domain of MinC inhibits assembly of the Z ring in Escherichia coli. AB - In Escherichia coli, the Min system, consisting of three proteins, MinC, MinD, and MinE, negatively regulates FtsZ assembly at the cell poles, helping to ensure that the Z ring will assemble only at midcell. Of the three Min proteins, MinC is sufficient to inhibit Z-ring assembly. By binding to MinD, which is mostly localized at the membrane near the cell poles, MinC is sequestered away from the cell midpoint, increasing the probability of Z-ring assembly there. Previously, it has been shown that the two halves of MinC have two distinct functions. The N terminal half is sufficient for inhibition of FtsZ assembly, whereas the C terminal half of the protein is required for binding to MinD as well as to a component of the division septum. In this study, we discovered that overproduction of the C-terminal half of MinC (MinC(122-231)) could also inhibit cell division and that this inhibition was at the level of Z-ring disassembly and dependent on MinD. We also found that fusing green fluorescent protein to either the N-terminal end of MinC(122-231), the C terminus of full-length MinC, or the C terminus of MinC(122-231) perturbed MinC function, which may explain why cell division inhibition by MinC(122-231) was not detected previously. These results suggest that the C-terminal half of MinC has an additional function in the regulation of Z-ring assembly. PMID- 17085578 TI - M.SpyI, a DNA methyltransferase encoded on a mefA chimeric element, modifies the genome of Streptococcus pyogenes. AB - While screening the clonality of Streptococcus pyogenes isolates from an outbreak of erythromycin-resistant pharyngitis in Pittsburgh, PA, we found a correlation between the presence of the chimeric element Phi10394.4 (carrying the macrolide efflux gene, mefA) and genomic DNA being resistant to cleavage by SmaI restriction endonuclease. A search of the open reading frames in Phi10394.4 identified a putative type II restriction-modification (R-M) cassette containing a cytosine methyltransferase gene (spyIM). Heterologous expression of the cloned spyIM gene, as well as allelic-replacement experiments, showed that the action of this methyltransferase (M.SpyI) was responsible for the inhibition of SmaI digestion of genomic DNA in the Phi10394.4-containing isolates. Analysis of the methylation patterns of streptococcal genomic DNA from spyIM-positive strains, a spyIM deletion mutant, and a spyIM-negative strain determined that M.SpyI specifically recognized and methylated the DNA sequence to generate 5'-C(m)CNGG. To our knowledge, this is the first methyltransferase gene from S. pyogenes to be cloned and to have its activity characterized. These results reveal why pulsed field gel electrophoresis analysis of SmaI-digested genomic DNA cannot be used to analyze the clonality of some streptococci containing Phi10394.4 and may explain the inability of previous epidemiological studies to use SmaI to analyze DNAs from macrolide-resistant streptococci. The presence of the SpyI R-M cassette in Phi10394.4 could impart a selective advantage to host strain survival and may provide another explanation for the observed increase in macrolide-resistant streptococci. PMID- 17085579 TI - Blood flow velocity effects and role of activation delay time on growth and form of platelet thrombi. AB - Mural thrombi are composed dominantly of platelets and develop under a blood flow. Portions can break off and are carried in the blood flow as emboli. Thrombus growth rates are affected by the velocity of the blood flow, but they do not simply increase with it, they exhibit a maximum, with subsequent decrease. Whereas this variation indicates an interaction of biochemical and physical processes, studies have concentrated widely on understanding only the biochemical processes. Here we show results of simulation of thrombus formation in 3D flows by accounting for the movements of individual platelets. Each platelet follows prescribed rules for interactions while the local flow around the thrombus continuously adjusts to the growing structure of the thrombus, also when embolization occurs. With an activation delay time assigned to each platelet we demonstrate the dependence of thrombus growth rate on blood velocity as found experimentally by Begent and Born [Begent N, Born GV (1970) Nature 227:926-930]. With activated platelets having mutual tensile action sustainable up to a prescribed distance we achieve thrombus growth faster than with shorter maximum distances that make a thrombus less porous; when the prescribed maximum distance is large enough the thrombus shape is not like a "hill" but like a "carpet." We find that thrombus growth rate is enhanced by modest pulsatility but less so when pulsations are amplified in part because of more embolization. PMID- 17085580 TI - A pivotal role for endogenous TGF-beta-activated kinase-1 in the LKB1/AMP activated protein kinase energy-sensor pathway. AB - TGF-beta-activated kinase-1 (TAK1), also known as MAPKK kinase-7 (MAP3K7), is a candidate effector of multiple circuits in cardiac biology and disease. Here, we show that inhibition of TAK1 in mice by a cardiac-specific dominant-negative mutation evokes electrophysiological and biochemical properties reminiscent of human Wolff-Parkinson-White syndrome, arising from mutations in AMP-activated protein kinase (AMPK), most notably, accelerated atrioventricular conduction and impaired AMPK activation. To test conclusively the biochemical connection from TAK1 to AMPK suggested by this phenotype, we disrupted TAK1 in mouse embryos and embryonic fibroblasts by Cre-mediated recombination. In TAK1-null embryos, the activating phosphorylation of AMPK at T172 was blocked, accompanied by defective AMPK activity. However, loss of endogenous TAK1 causes midgestation lethality, with defective yolk sac and intraembryonic vasculature. To preclude confounding lethal defects, we acutely ablated floxed TAK1 in culture by viral delivery of Cre. In culture, endogenous TAK1 was activated by oligomycin, the antidiabetic drug metformin, 5-aminoimidazole-4-carboxamide riboside (AICAR), and ischemia, well established triggers of AMPK activity. Loss of TAK1 in culture blocked T172 phosphorylation induced by all three agents, interfered with AMPK activation, impaired phosphorylation of the endogenous AMPK substrate acetyl CoA carboxylase, and also interfered with activation of the AMPK kinase LKB1. Thus, by disrupting the endogenous TAK1 locus, we prove a pivotal role for TAK1 in the LKB1/AMPK signaling axis, an essential governor of cell metabolism. PMID- 17085581 TI - Biogenesis of actin-like bacterial cytoskeletal filaments destined for positioning prokaryotic magnetic organelles. AB - Magnetosomes comprise a magnetic nanocrystal surrounded by a lipid bilayer membrane. These unique prokaryotic organelles align inside magnetotactic bacterial cells and serve as an intracellular compass allowing the bacteria to navigate along the geomagnetic field in aquatic environments. Cryoelectron tomography of Magnetospirillum strains has revealed that the magnetosome chain is surrounded by a network of filaments that may be composed of MamK given that the filaments are absent in the mamK mutant cells. The process of the MamK filament assembly is unknown. Here we prove the authenticity of the MamK filaments and show that MamK exhibits linear distribution inside Magnetospirillum sp. cells even in the area without magnetosomes. The mamK gene alone is sufficient to direct the synthesis of straight filaments in Escherichia coli, and one extremity of the MamK filaments is located at the cellular pole. By using dual fluorescent labeling of MamK, we found that MamK nucleates at multiple sites and assembles into mosaic filaments. Time-lapse experiments reveal that the assembly of the MamK filaments is a highly dynamic and kinetically asymmetrical process. MamK bundles might initiate the formation of a new filament or associate to one preexistent filament. Our results demonstrate the mechanism of biogenesis of prokaryotic cytoskeletal filaments that are structurally and functionally distinct from the known MreB and ParM filaments. In addition to positioning magnetosomes, other hypothetical functions of the MamK filaments in magnetotaxis might include anchoring magnetosomes and being involved in magnetic reception. PMID- 17085582 TI - A proboscidean from the late Oligocene of Eritrea, a "missing link" between early Elephantiformes and Elephantimorpha, and biogeographic implications. AB - We report on a late Oligocene proboscidean species from Eritrea, dated to 26.8 +/ 1.5 Mya. This "missing link" between early elephantiformes and Elephantimorpha is the oldest known nongomphothere proboscidean to probably display horizontal tooth displacement, typical of elephants [Elephantimorpha consists of Mammutida (mastodons) and Elephantida, and Elephantida includes gomphotheres, stegodons, and elephants]. Together with the newly discovered late Oligocene gomphotheres from Chilga, Ethiopia, the Eritrean taxon points to the importance of East Africa as a major area for the knowledge of the early evolution of Elephantimorpha before the faunal exchange between Eurasia and Africa. PMID- 17085583 TI - Flavodoxin hydroquinone reduces Azotobacter vinelandii Fe protein to the all ferrous redox state with a S = 0 spin state. AB - Azotobacter vinelandii flavodoxin hydroquinone (FldHQ) is a physiological reductant to nitrogenase supporting catalysis that is twice as energy efficient (ATP/2e- = 2) as dithionite (ATP/2e- = 4). This catalytic efficiency results from reduction of Fe protein from A. vinelandii (Av2) to the all-ferrous oxidation state ([Fe4S4]0), in contrast to dithionite, which only reduces Av2 to the [Fe4S4]1+ state. Like FldHQ, Ti(III) citrate yields ATP/2e- = 2, and Ti(III) reduced [Fe4S4]0 Av2 has a S = 4 spin state and characteristic Mossbauer spectrum, a parallel mode g = 16.4 EPR signal, and a shoulder at 520 nm in its UV vis spectrum, each of which distinguish the S = 4 [Fe4S4]0 Av2 from other states. In this study, we demonstrate that FldHQ makes [Fe4S4]0 Av2, which is sufficiently characterized to demonstrate unique physical properties that distinguish it from the previously characterized Ti(III)-reduced [Fe4S4]0 Av2. In particular, Evans NMR magnetic susceptibility and EPR measurements indicate that FldHQ-reduced [Fe4S4]0 Av2 has an S = 0 spin state (like [Fe4S4]2+ Av2). There is no g = 16.4 EPR signal and no shoulder at 520 nm in its absorbance spectrum, which resembles that of [Fe4S4]1+ Av2. That the physiological reductant to Av2 is capable of forming [Fe4S4]0 Av2 has important implications for in vivo nitrogenase activity. PMID- 17085584 TI - Rapidly evolving zona pellucida domain proteins are a major component of the vitelline envelope of abalone eggs. AB - Proteins harboring a zona pellucida (ZP) domain are prominent components of vertebrate egg coats. Although less well characterized, the egg coat of the non vertebrate marine gastropod abalone (Haliotis spp.) is also known to contain a ZP domain protein, raising the possibility of a common molecular basis of metazoan egg coat structures. Egg coat proteins from vertebrate as well as non-vertebrate taxa have been shown to evolve under positive selection. Studied most extensively in the abalone system, coevolution between adaptively diverging egg coat and sperm proteins may contribute to the rapid development of reproductive isolation. Thus, identifying the pattern of evolution among egg coat proteins is important in understanding the role these genes may play in the speciation process. The purpose of the present study is to characterize the constituent proteins of the egg coat [vitelline envelope (VE)] of abalone eggs and to provide preliminary evidence regarding how selection has acted on VE proteins during abalone evolution. A proteomic approach is used to match tandem mass spectra of peptides from purified VE proteins with abalone ovary EST sequences, identifying 9 of 10 ZP domain proteins as components of the VE. Maximum likelihood models of codon evolution suggest positive selection has acted among a subset of amino acids for 6 of these genes. This work provides further evidence of the prominence of ZP proteins as constituents of the egg coat, as well as the prominent role of positive selection in diversification of these reproductive proteins. PMID- 17085585 TI - Cis-urocanic acid, a sunlight-induced immunosuppressive factor, activates immune suppression via the 5-HT2A receptor. AB - Exposure to UV radiation induces skin cancer and suppresses the immune response. To induce immune suppression, the electromagnetic energy of UV radiation must be absorbed by an epidermal photoreceptor and converted into a biologically recognizable signal. Two photoreceptors have been recognized: DNA and trans urocanic acid (UCA). Trans-UCA is normally found in the outermost layer of skin and isomerizes to the cis isomer upon exposure to UV radiation. Although UCA was identified as a UV photoreceptor years ago, and many have documented its ability to induce immune suppression, its exact mode of action remains elusive. Particularly vexing has been the identity of the molecular pathway by which cis UCA mediates immune suppression. Here we provide evidence that cis-UCA binds to the serotonin [5-hydroxytryptamine (5-HT)] receptor with relatively high affinity (Kd = 4.6 nM). Anti-cis-UCA antibody precipitates radiolabeled 5-HT, and the binding is inhibited by excess 5-HT and/or excess cis-UCA. Similarly, anti-5-HT antibody precipitates radiolabeled cis-UCA, and the binding is inhibited by excess 5-HT or excess cis-UCA. Calcium mobilization was activated when a mouse fibroblast line, stably transfected with the human 5-HT2A receptor, was treated with cis-UCA. Cis-UCA-induced calcium mobilization was blocked with a selective 5 HT2A receptor antagonist. UV- and cis-UCA-induced immune suppression was blocked by antiserotonin antibodies or by treating the mice with 5-HT2A receptor antagonists. Our findings identify cis-UCA as a serotonin receptor ligand and indicate that the immunosuppressive effects of cis-UCA and UV radiation are mediated by activation of the 5-HT2A receptor. PMID- 17085586 TI - Catalytic reduction of dinitrogen to ammonia at a single molybdenum center. AB - Since our discovery of the catalytic reduction of dinitrogen to ammonia at a single molybdenum center, we have embarked on a variety of studies designed to further understand this complex reaction cycle. These include studies of both individual reaction steps and of ligand variations. An important step in the reaction sequence is exchange of ammonia for dinitrogen in neutral molybdenum(III) compounds. We have found that this exchange reaction is first order in dinitrogen and relatively fast (complete in <1 h) at 1 atm of dinitrogen. Variations of the terphenyl substituents in the triamidoamine ligand demonstrate that the original ligand is not unique in its ability to yield successful catalysts. However, complexes that contain sterically less demanding ligands fail to catalyze formation of ammonia from dinitrogen; it is proposed as a consequence of a base-catalyzed decomposition of a diazenido (Mo-N=NH) intermediate. PMID- 17085587 TI - Source reduction of mosquito larval habitats has unexpected consequences on malaria transmission. AB - Reduction of aquatic habitats through environmental management mitigates malaria transmission not only by reducing emergence of host-seeking mosquitoes, but also by increasing the amount of time required for vectors to locate oviposition sites. However, the consequence of source reduction on mosquito oviposition has largely been neglected in evaluations of environment-management programs. Here, by theoretically examining the relationship between the time spent for oviposition and the availability of aquatic habitats, we show that prolonged oviposition cycles induced by source reduction account for a great deal of reductions in the basic reproductive rate of malaria, especially when aquatic habitats are scarce and the mosquito's flight ability is limited. Neglecting this mechanism may lead to substantial underestimation of the impact of source reduction of aquatic habitats on malaria transmission. Our findings suggest that the prolonged duration of the gonotrophic cycle might be one of the important mechanisms underlying the effectiveness of environment-management interventions for malaria control. PMID- 17085588 TI - Early modern humans from the Pestera Muierii, Baia de Fier, Romania. AB - The early modern human remains from the Pestera Muierii, Romania have been directly dated to approximately 30,000 radiocarbon years before present (approximately 30 ka 14C BP) (approximately 35 ka cal BP) ("calendrical" age; based on CalPal 2005) and augment a small sample of securely dated, European, pre 28 ka 14C BP (approximately 32.5 ka cal BP) modern human remains. The Muierii fossils exhibit a suite of derived modern human features, including reduced maxillae with pronounced canine fossae, a narrow nasal aperture, small superciliary arches, an arched parietal curve, zygomatic arch above the auditory porous, laterally bulbous mastoid processes, narrow mandibular corpus, reduced anterior dentition, ventral-to-bisulcate scapular axillary border, and planoconcave tibial and fibular diaphyseal surfaces. However, these traits co occur with contextually archaic and/or Neandertal features, including a moderately low frontal arc, a large occipital bun, a high coronoid process and asymmetrical mandibular notch, a more medial mandibular notch crest to condylar position, and a narrow scapular glenoid fossa. As with other European early modern humans, the mosaic of modern human and archaic/Neandertal features, relative to their potential Middle Paleolithic ancestral populations, indicates considerable Neandertal/modern human admixture. Moreover, the narrow scapular glenoid fossa suggests habitual movements at variance with the associated projectile technology. The reproductive and scapulohumeral functional inferences emphasize the subtle natures of behavioral contrasts between Neandertals and these early modern Europeans. PMID- 17085589 TI - Feasibility of cooling the Earth with a cloud of small spacecraft near the inner Lagrange point (L1). AB - If it were to become apparent that dangerous changes in global climate were inevitable, despite greenhouse gas controls, active methods to cool the Earth on an emergency basis might be desirable. The concept considered here is to block 1.8% of the solar flux with a space sunshade orbited near the inner Lagrange point (L1), in-line between the Earth and sun. Following the work of J. Early [Early, JT (1989) J Br Interplanet Soc 42:567-569], transparent material would be used to deflect the sunlight, rather than to absorb it, to minimize the shift in balance out from L1 caused by radiation pressure. Three advances aimed at practical implementation are presented. First is an optical design for a very thin refractive screen with low reflectivity, leading to a total sunshade mass of approximately 20 million tons. Second is a concept aimed at reducing transportation cost to 50 dollars/kg by using electromagnetic acceleration to escape Earth's gravity, followed by ion propulsion. Third is an implementation of the sunshade as a cloud of many spacecraft, autonomously stabilized by modulating solar radiation pressure. These meter-sized "flyers" would be assembled completely before launch, avoiding any need for construction or unfolding in space. They would weigh a gram each, be launched in stacks of 800,000, and remain for a projected lifetime of 50 years within a 100,000-km-long cloud. The concept builds on existing technologies. It seems feasible that it could be developed and deployed in approximately 25 years at a cost of a few trillion dollars, <0.5% of world gross domestic product (GDP) over that time. PMID- 17085590 TI - SK channels mediate NADPH oxidase-independent reactive oxygen species production and apoptosis in granulocytes. AB - Neutrophils are immune cells that bind to, engulf, and destroy bacterial and fungal pathogens in infected tissue, and their clearance by apoptosis is essential for the resolution of inflammation. Killing involves both oxidative and nonoxidative processes, the oxidative pathway requiring electrogenic production of superoxide by the membrane-bound NADPH oxidase complex. A variety of stimuli, from bacterial chemotactic peptides to complement- or IgG-opsonized microbes, can induce the production of reactive oxygen species (ROS) by neutrophils, presumably by means of NADPH oxidase. We report here that 1-ethyl-2-benzimidazolinone (1 EBIO), an activator of Ca2+-activated potassium channels of small conductance (SK) and intermediate conductance (IK), causes production of superoxide and hydrogen peroxide by neutrophils and granulocyte-differentiated PLB-985 cells. This response can be partially inhibited by the SK blocker apamin, which inhibits a Ca2+-activated K+ current in these cells. Analysis of RNA transcripts indicates that channels encoded by the SK3 gene carry this current. The effects of 1-EBIO and apamin are independent of the NADPH oxidase pathway, as demonstrated by using a PLB-985 cell line lacking the gp91phox subunit. Rather, 1-EBIO and apamin modulate mitochondrial ROS production. Consistent with the enhanced ROS production and K+ efflux mediated by 1-EBIO, we found that this SK opener increased apoptosis of PLB-985 cells. Together, these findings suggest a previously uncharacterized mechanism for the regulation of neutrophil ROS production and programmed cell death. PMID- 17085591 TI - Insights into transcription enhancer factor 1 (TEF-1) activity from the solution structure of the TEA domain. AB - Transcription enhancer factor 1 is essential for cardiac, skeletal, and smooth muscle development and uses its N-terminal TEA domain (TEAD) to bind M-CAT elements. Here, we present the first structure of TEAD and show that it is a three-helix bundle with a homeodomain fold. Structural data reveal how TEAD binds DNA. Using structure-function correlations, we find that the L1 loop is essential for cooperative loading of TEAD molecules on to tandemly duplicated M-CAT sites. Furthermore, using a microarray chip-based assay, we establish that known binding sites of the full-length protein are only a subset of DNA elements recognized by TEAD. Our results provide a model for understanding the regulation of genome-wide gene expression during development by TEA/ATTS family of transcription factors. PMID- 17085592 TI - Small dsRNAs induce transcriptional activation in human cells. AB - Recent studies have shown that small noncoding RNAs, such as microRNAs and siRNAs, regulate gene expression at multiple levels including chromatin architecture, transcription, RNA editing, RNA stability, and translation. Each form of RNA-dependent regulation has been generally found to silence homologous sequences and collectively called RNAi. To further study the regulatory role of small RNAs at the transcriptional level, we designed and synthesized 21-nt dsRNAs targeting selected promoter regions of human genes E-cadherin, p21(WAF1/CIP1) (p21), and VEGF. Surprisingly, transfection of these dsRNAs into human cell lines caused long-lasting and sequence-specific induction of targeted genes. dsRNA mutation studies reveal that the 5' end of the antisense strand, or "seed" sequence, is critical for activity. Mechanistically, the dsRNA-induced gene activation requires the Argonaute 2 (Ago2) protein and is associated with a loss of lysine-9 methylation on histone 3 at dsRNA-target sites. In conclusion, we have identified several dsRNAs that activate gene expression by targeting noncoding regulatory regions in gene promoters. These findings reveal a more diverse role for small RNA molecules in the regulation of gene expression than previously recognized and identify a potential therapeutic use for dsRNA in targeted gene activation. PMID- 17085593 TI - Two modes of microtubule sliding driven by cytoplasmic dynein. AB - Dynein is a huge multisubunit microtubule (MT)-based motor, whose motor domain resides in the heavy chain. The heavy chain comprises a ring of six AAA (ATPases associated with diverse cellular activities) modules with two slender protruding domains, the tail and stalk. It has been proposed that during the ATP hydrolysis cycle, this tail domain swings against the AAA ring as a lever arm to generate the power stroke. However, there is currently no direct evidence to support the model that the tail swing is tightly linked to dynein motility. To address the question of whether the power stroke of the tail drives MT sliding, we devised an in vitro motility assay using genetically biotinylated cytoplasmic dyneins anchored on a glass surface in the desired orientation with a biotin-streptavidin linkage. Assays on the dyneins with the site-directed biotin tag at eight different locations provided evidence that robust MT sliding is driven by the power stroke of the tail. Furthermore, the assays revealed slow MT sliding independent of dynein orientation on the glass surface, which is mechanically distinct from the sliding driven by the power stroke of the tail. PMID- 17085594 TI - Quantitative multispectral biosensing and 1D imaging using quasi-3D plasmonic crystals. AB - We developed a class of quasi-3D plasmonic crystal that consists of multilayered, regular arrays of subwavelength metal nanostructures. The complex, highly sensitive structure of the optical transmission spectra of these crystals makes them especially well suited for sensing applications. Coupled with quantitative electrodynamics modeling of their optical response, they enable full multiwavelength spectroscopic detection of molecular binding events with sensitivities that correspond to small fractions of a monolayer. The high degree of spatial uniformity of the crystals, formed by a soft nanoimprint technique, provides the ability to image binding events over large areas with micrometer spatial resolution. These features, together with compact form factors, low-cost fabrication procedures, simple readout apparatus, and ability for direct integration into microfluidic networks and arrays, suggest promise for these devices in label-free bioanalytical detection systems. PMID- 17085595 TI - Recent history of artificial outcrossing facilitates whole-genome association mapping in elite inbred crop varieties. AB - Genomewide association studies depend on the extent of linkage disequilibrium (LD), the number and distribution of markers, and the underlying structure in populations under study. Outbreeding species generally exhibit limited LD, and consequently, a very large number of markers are required for effective whole genome association genetic scans. In contrast, several of the world's major food crops are self-fertilizing inbreeding species with narrow genetic bases and theoretically extensive LD. Together these are predicted to result in a combination of low resolution and a high frequency of spurious associations in LD based studies. However, inbred elite plant varieties represent a unique human induced pseudo-outbreeding population that has been subjected to strong selection for advantageous alleles. By assaying 1,524 genomewide SNPs we demonstrate that, after accounting for population substructure, the level of LD exhibited in elite northwest European barley, a typical inbred cereal crop, can be effectively exploited to map traits by using whole-genome association scans with several hundred to thousands of biallelic SNPs. PMID- 17085596 TI - Discovery and in vitro biosynthesis of haloduracin, a two-component lantibiotic. AB - Lantibiotics are ribosomally synthesized peptides that undergo posttranslational modifications to their mature, antimicrobial form. They are characterized by the unique amino acids lanthionine and methyllanthionine, introduced by means of dehydration of Ser/Thr residues followed by reaction of the resulting dehydro amino acids with cysteines to form thioether linkages. Two-component lantibiotics use two peptides that are each posttranslationally modified to yield two functionally distinct products that act in synergy to provide bactericidal activity. By using genetic data instead of isolation, a two-component lantibiotic, haloduracin, was identified in the genome of the Gram-positive alkaliphilic bacterium Bacillus halodurans C-125. We show that heterologously expressed and purified precursor peptides HalA1 and HalA2 are processed by the purified modification enzymes HalM1 and HalM2 in an in vitro reconstitution of the biosynthesis of a two-component lantibiotic. The activity of each HalM enzyme is substrate-specific, and the assay products exhibit antimicrobial activity after removal of their leader sequences at an engineered Factor Xa cleavage site, indicating that correct thioether formation has occurred. Haloduracin's biological activity depends on the presence of both modified peptides. The structures of the two mature haloduracin peptides Halalpha and Halbeta were investigated, indicating that they have similarities as well as some distinct differences compared with other two-component lantibiotics. PMID- 17085597 TI - Structural basis for protein-protein interactions in the 14-3-3 protein family. AB - The seven members of the human 14-3-3 protein family regulate a diverse range of cell signaling pathways by formation of protein-protein complexes with signaling proteins that contain phosphorylated Ser/Thr residues within specific sequence motifs. Previously, crystal structures of three 14-3-3 isoforms (zeta, sigma, and tau) have been reported, with structural data for two isoforms deposited in the Protein Data Bank (zeta and sigma). In this study, we provide structural detail for five 14-3-3 isoforms bound to ligands, providing structural coverage for all isoforms of a human protein family. A comparative structural analysis of the seven 14-3-3 proteins revealed specificity determinants for binding of phosphopeptides in a specific orientation, target domain interaction surfaces and flexible adaptation of 14-3-3 proteins through domain movements. Specifically, the structures of the beta isoform in its apo and peptide bound forms showed that its binding site can exhibit structural flexibility to facilitate binding of its protein and peptide partners. In addition, the complex of 14-3-3 beta with the exoenzyme S peptide displayed a secondary structural element in the 14-3-3 peptide binding groove. These results show that the 14-3-3 proteins are adaptable structures in which internal flexibility is likely to facilitate recognition and binding of their interaction partners. PMID- 17085598 TI - Telomere-mediated chromosomal truncation in maize. AB - Direct repeats of Arabidopsis telomeric sequence were constructed to test telomere-mediated chromosomal truncation in maize. Two constructs with 2.6 kb of telomeric sequence were used to transform maize immature embryos by Agrobacterium mediated transformation. One hundred seventy-six transgenic lines were recovered in which 231 transgene loci were revealed by a FISH analysis. To analyze chromosomal truncations that result in transgenes located near chromosomal termini, Southern hybridization analyses were performed. A pattern of smear in truncated lines was seen as compared with discrete bands for internal integrations, because telomeres in different cells are elongated differently by telomerase. When multiple restriction enzymes were used to map the transgene positions, the size of the smears shifted in accordance with the locations of restriction sites on the construct. This result demonstrated that the transgene was present at the end of the chromosome immediately before the integrated telomere sequence. Direct evidence for chromosomal truncation came from the results of FISH karyotyping, which revealed broken chromosomes with transgene signals at the ends. These results demonstrate that telomere-mediated chromosomal truncation operates in plant species. This technology will be useful for chromosomal engineering in maize as well as other plant species. PMID- 17085599 TI - Dual recognition of herpes simplex viruses by TLR2 and TLR9 in dendritic cells. AB - Dendritic cells (DCs) express multiple Toll-like receptors (TLR) in distinct cellular locations. Herpes simplex viruses (HSV) have been reported to engage both the surface TLR2 and intracellular TLR9 in conventional DCs. However, the contributions of these TLRs in recognition of HSV and the induction of proinflammatory cytokines in DCs remain unclear. Here, we demonstrate that a rare population of HSV, both in laboratory strains and in primary clinical isolates from humans, has the capacity to activate TLR2. This virus population is recognized through both TLR2 and TLR9 for the induction of IL-6 and IL-12 secretion from bone marrow-derived DCs. Further, we describe a previously uncharacterized pathway of viral recognition in which TLR2 and TLR9 are engaged in sequence within the same DC. Live viral infection results in two additional agonists of TLR2 and TLR9. These results indicate that in cells that express multiple TLRs, pathogens that contain multiple pathogen-associated molecular patterns can be detected in an orchestrated sequence and suggest that the innate immune system in DCs is optimized to linking uptake and degradation of pathogens to microbial recognition. PMID- 17085600 TI - An exceptionally fast actomyosin reaction powers insect flight muscle. AB - Insects, as a group, have been remarkably successful in adapting to a great range of physical and biological environments, in large part because of their ability to fly. The evolution of flight in small insects was accompanied by striking adaptations of the thoracic musculature that enabled very high wing beat frequencies. At the cellular and protein filament level, a stretch activation mechanism evolved that allowed high-oscillatory work to be achieved at very high frequencies as contraction and nerve stimulus became asynchronous. At the molecular level, critical adaptations occurred within the motor protein myosin II, because its elementary interactions with actin set the speed of sarcomere contraction. Here, we show that the key myosin enzymatic adaptations required for powering the very fast flight muscles in the fruit fly Drosophila melanogaster include the highest measured detachment rate of myosin from actin (forward rate constant, 3,698 s(-1)), an exceptionally weak affinity of MgATP for myosin (association constant, 0.2 mM(-1)), and a unique rate-limiting step in the cross bridge cycle at the point of inorganic phosphate release. The latter adaptations are constraints imposed by the overriding requirement for exceptionally fast release of the hydrolytic product MgADP. Otherwise, as in Drosophila embryonic muscle and other slow muscle types, a step associated with MgADP release limits muscle contraction speed by delaying the detachment of myosin from actin. PMID- 17085601 TI - Conditional hospital release: conveying the wrong message? PMID- 17085602 TI - Innovations: evidence-based practices: establishing the evidence base for psychiatric services: estimating the impact on the population. AB - The impact of evidence-based interventions for individual patients is clear, but a broader approach is needed. The authors describe a six-step method for calculating the population impact of introducing a new evidence-based intervention. The steps involve identifying the prevalence of a condition, the proportion of persons in the larger population eligible for the intervention, and the baseline risk that the intervention is meant to address. Obtaining useful results from this mathematical calculation depends on having accurate and robust data about the prevalence of a condition, the size of the targeted population, and the effectiveness of the intervention. Results for different interventions for the same condition and target population can then be compared. PMID- 17085603 TI - State mental health policy: statewide implementation of the crisis intervention team program: the Ohio model. AB - This column discusses ways that states can implement community-based best practices statewide, by using the crisis intervention team (CIT) model as an example. Although state mental health authorities may want to use a top-down approach to ensure uniform, high-quality implementation, programs may be more likely to succeed if they arise as bottom-up, grassroots innovations. Programs like CIT are especially challenging to implement because they involve collaboration between complex systems and affect multiple stakeholders. The column describes lessons learned in Ohio in hopes of assisting other states in implementing this and other innovations. PMID- 17085604 TI - Characteristics of U.S. youths with serious emotional disturbance: data from the National Health Interview Survey. AB - OBJECTIVE: Although it is estimated that serious emotional disturbance affects 9 to 13 percent of children and adolescents in the United States, there are few national data on the characteristics of this group. METHODS: This study used data for 13,579 youths from the 2001 National Health Interview Survey (NHIS) to describe the sociodemographic features and insurance coverage of youths with serious emotional disturbance living in the United States. Youths with serious emotional disturbance were identified through their scores on the Strengths and Difficulties Questionnaire, which was added to the NHIS in 2001. RESULTS: A large majority of youths with serious emotional disturbance were white and had income at 200 percent of the poverty level or higher. About 40 percent of youths with serious emotional disturbance had private insurance coverage, whereas Medicaid and the State Children's Health Insurance Program provided coverage for about a third of youths with serious emotional disturbance. CONCLUSIONS: Although Medicaid is an important payer of mental health services for youths with serious emotional disturbance, private insurance is still the primary source of health coverage for youths with serious emotional disturbance and for the overall population of youths. PMID- 17085605 TI - Building on practice-based evidence: using expert perspectives to define the wraparound process. AB - OBJECTIVE: In order to expand the research base on effective community-based mental health treatments, methods are needed to define and evaluate promising interventions that have not been systematically developed and tested. In this report, the authors describe the results of an effort to better define the wraparound process for children and adolescents with serious emotional and behavioral problems. METHODS: A broad review of wraparound treatment manuals and model descriptions was conducted. With the help of a small group of experts, this review was synthesized into an initial description of the phases and activities of the wraparound process. This model was then presented to a multidisciplinary advisory panel of 31 experts on the wraparound process who provided structured and semistructured feedback. RESULTS: Overall, respondents expressed a high level of agreement with the proposed set of activities. For 23 of the 31 activities presented, there was unanimous or near-unanimous agreement (that is, one dissenter) that the activity was an essential component of the wraparound process. For 20 of the 31 activities, there was unanimous agreement that the description was phrased acceptably. A final model was created on the basis of feedback from reviewers. CONCLUSIONS: Results indicate that using the experience of a wide base of stakeholders to operationalize a complex model such as wraparound is feasible and holds many potential benefits, including building consensus in the field, improving service quality, and accelerating the incorporation of evaluation results into real-world practice. PMID- 17085606 TI - Relations between program and system variables and fidelity to the wraparound process for children and families. AB - OBJECTIVE: Past research and experience have indicated that characteristics of organizations and service systems influence the quality of mental health services delivery. The study aimed to illuminate such associations by examining the relationship between a set of program- and system-level variables and fidelity to the wraparound model for children and families. METHODS: A measure of fidelity to the wraparound process for children and families (the Wraparound Fidelity Index) was administered to families receiving services in eight mental health agencies across the United States. Program administrators at each agency also completed by telephone a standardized interview for assessing program and system conditions hypothesized to influence quality of implementation (Program Administrator version of the Wraparound Fidelity Index). Regression analyses were used to assess the relationship between program and system variables and fidelity. RESULTS: Results of interviews with program administrators revealed substantial variation across programs in the number and type of program and system supports for wraparound implementation. Regression analyses found a significant association between the number of organizational and system supports and Wraparound Fidelity Index scores. CONCLUSIONS: Results support the hypothesized relationship between program and system conditions and the quality of service delivery. Results also highlight the importance of engineering the system and organizational context (maintaining low caseload sizes, establishing interagency partnerships, and implementing accountability mechanisms) in order to provide a solid foundation for high-quality wraparound to children and families. PMID- 17085607 TI - Within-state availability of transition-to-adulthood services for youths with serious mental health conditions. AB - OBJECTIVE: This study describes the existence and nature of services within state child and adult mental health systems that support the transition from adolescence to adulthood. METHODS: State child and adult mental health administrators from all but one state were interviewed by telephone with a semistructured questionnaire regarding transition services in their state mental health system, such as supported housing, vocational support, preparation for independent living, and dual diagnosis treatment. Eight states were deemed sufficiently decentralized to render state-level administrator reports invalid. Specific service data from the remaining 41 states and the District of Columbia were analyzed with descriptive statistics. RESULTS: One-quarter of child state mental health systems and one-half of adult state mental health systems offered no transition services, and few provided any kind of transition service at more than one site. Most types of transition services were available at all in less than 20 percent of the states. CONCLUSIONS: Across the United States transition support services are lacking. The adult system in particular will require major transformation to provide the service capacity that is needed to meet the current standards of transition service accessibility for young Americans with serious mental health conditions. PMID- 17085608 TI - Conditional release: a less restrictive alternative to hospitalization? AB - OBJECTIVE: This study examined conditional release--that is, involuntary outpatient commitment orders upon release from hospitalization--as a least restrictive alternative to psychiatric hospitalization in Victoria, Australia. METHODS: Records were obtained from the Victorian Psychiatric Case Register for patients who experienced psychiatric hospitalization: between 1990 and 2000 a total of 8,879 patients were given conditional release and 16,094 were not. RESULTS: Compared with the group that was hospitalized but did not receive a conditional release, the group that received a conditional release was more likely to have more prior hospitalizations of greater than average duration. Patients with schizophrenia were more likely to be given conditional release. Patients given conditional release experienced a care pattern involving briefer inpatient episodes (8.3 fewer days per episode), more inpatient days, and longer duration of restrictive care--that is, combined inpatient and conditional release periods (5.1 more days per month in care). CONCLUSIONS: For patients at risk of long-term hospitalization, conditional release may help to shorten inpatient episodes by providing a least restrictive alternative to continued hospitalization. However, patients who were given conditional release doubled the amount of days they spent under restrictive care, compared with the amount of time they previously spent in the hospital before entering a period of combined inpatient and conditional release commitment. Additional oversight may have led to more frequent hospitalization. This consequence raises new questions regarding the possible benefits of such extended oversight and new challenges for release planning using conditional release as a least restrictive method of care. PMID- 17085609 TI - Effect of conditional release from hospitalization on mortality risk. AB - OBJECTIVES: This study considered the protective value provided by conditional release. It assessed the contribution of conditional release to mortality risk among patients with mental disorders severe enough to require psychiatric hospitalization during a mental health treatment span of 13.5 years in Victoria, Australia. METHODS: Death records were obtained from the Australian National Death Index for a sample of 24,973 Victorian Psychiatric Case Register patients with a history of psychiatric hospitalizations: 8,879 had experienced at least one conditional release during community care intervals and 16,094 had not. Risk of death was assessed with standardized mortality ratios of the general population of Victoria. Relative risk of death among patients with and without past experience of conditional release was computed with risk and odds ratios. The contribution of conditional release to mortality, taking into account use of community care services, age, gender, inpatient experience, and diagnosis, as well as other controls, was assessed with logistic regression. RESULTS: Patients who had been hospitalized showed higher mortality risk than the general population. Sixteen percent (4,034) died. Patients exposed to conditional release, however, had a 14 percent reduction in probability of non-injury-related death and a 24 percent reduction per day on orders in the probability of death from injury compared with those not offered such oversight throughout their mental health treatment, all other factors taken into account. CONCLUSIONS: Conditional release can offer protective oversight for those considered dangerous to self or others and appears to reduce mortality risk among those with disorders severe enough to require psychiatric hospitalization. PMID- 17085610 TI - Factors in the selection of patients for conditional release from their first psychiatric hospitalization. AB - OBJECTIVE: This study examined a sample of patients in Victoria, Australia, to identify factors in selection for conditional release from an initial hospitalization that occurred within 30 days of entry into the mental health system. METHODS: Data were from the Victorian Psychiatric Case Register. All patients first hospitalized and conditionally released between 1990 and 2000 were identified (N=8,879), and three comparison groups were created. Two groups were hospitalized within 30 days of entering the system: those who were given conditional release and those who were not. A third group was conditionally released from a hospitalization that occurred after or extended beyond 30 days after system entry. Logistic regression identified characteristics that distinguished the first group. Ordinary least-squares regression was used to evaluate the contribution of conditional release early in treatment to reducing inpatient episodes, inpatient days, days per episode, and inpatient days per 30 days in the system. RESULTS: Conditional release early in treatment was used for 11 percent of the sample, or more than a third of those who were eligible for this intervention. Factors significantly associated with selection for early conditional release were those related to a better prognosis (initial hospitalization at a later age and having greater than an 11th grade education), a lower likelihood of a diagnosis of dementia or schizophrenia, involuntary status at first inpatient admission, and greater community involvement (being employed and being married). When the analyses controlled for these factors, use of conditional release early in treatment was significantly associated with a reduction in use of subsequent inpatient care. PMID- 17085611 TI - Patterns and prevalence of arrest in a statewide cohort of mental health care consumers. AB - OBJECTIVE: Although criminal justice involvement among persons with severe mental illness is a much discussed topic, few large-scale studies systematically describe the patterns and prevalence of arrest in this population. This study examined rates, patterns, offenses, and sociodemographic correlates of arrest in a large cohort of mental health service recipients. METHODS: The arrest records of 13,816 individuals receiving services from the Massachusetts Department of Mental Health from 1991 to 1992 were examined over roughly a ten-year period. Bivariate relationships between sociodemographic factors and arrest were also examined. RESULTS: About 28 percent of the cohort experienced at least one arrest. The most common charges were crimes against public order followed by serious violent offenses and minor property crime. The number of arrests per individual ranged from one to 71. Five percent of arrestees (roughly 1.5 percent of the cohort) accounted for roughly 17 percent of arrests. The proportion of men arrested was double that of women. Persons 18 to 25 years of age had a 50 percent chance of at least one arrest. This rate declined with age but did so unevenly across offense types. CONCLUSIONS: The likelihood of arrest appeared substantial among persons with severe mental illness, but the bulk of offending appeared concentrated in a small group of persons and among persons with sociodemographic features similar to those of offenders in the general population. Data such as these could provide a platform for designing jail diversion and other services to reduce both initial and repeat offending among persons with serious mental illness. PMID- 17085612 TI - Assessments by patients with schizophrenia and psychiatrists of relative risk of research procedures. AB - OBJECTIVE: Schizophrenia studies involve diverse procedures with varying levels of risk. Federal regulations indicate that oversight of these protocols entails assessment of risk in relation to the risks encountered in everyday life. No data exist on comparing assessments of people with schizophrenia with those of psychiatrists regarding research procedure risks in relation to the usual risks of living with schizophrenia. METHODS: Structured interviews were given to 43 people with schizophrenia and a parallel written survey was given to 68 psychiatrists to compare assessments of relative risk. Twelve research procedures were rated to compare the risk of the procedure with everyday risks. Possible scores range from 1, much less risk, to 3, about the same, to 5, much more risk. RESULTS: People with schizophrenia and psychiatrists viewed filling out questionnaires, drawing a tube of blood, drawing a tube of blood for a genetic test, and having X-rays of the head as being less risky than usual daily risks. Psychiatrists saw showing a word or picture that is upsetting as being less risky than usual daily risks. In contrast, patients saw having a spinal tap, getting a new experimental medication, getting a medication that causes temporary symptoms of schizophrenia, and stopping usual medications for two weeks as being significantly more risky than usual daily risks, but psychiatrists rated only stopping usual medications for two weeks as being more risky than usual daily risks. CONCLUSIONS: Substantial congruence exists between patients and psychiatrists regarding their views of research procedure risks. Further work is needed to determine how these assessments may be used in evaluating protocols. PMID- 17085613 TI - Shared decision making and medication management in the recovery process. AB - Mental health professionals commonly conceptualize medication management for people with severe mental illness in terms of strategies to increase compliance or adherence. The authors argue that compliance is an inadequate construct because it fails to capture the dynamic complexity of autonomous clients who must navigate decisional conflicts in learning to manage disorders over the course of years or decades. Compliance is rooted in medical paternalism and is at odds with principles of person-centered care and evidence-based medicine. Using medication is an active process that involves complex decision making and a chance to work through decisional conflicts. It requires a partnership between two experts: the client and the practitioner. Shared decision making provides a model for them to assess a treatment's advantages and disadvantages within the context of recovering a life after a diagnosis of a major mental disorder. PMID- 17085614 TI - The tipping point from private practice to publicly funded settings for early- and mid-career psychiatrists. AB - OBJECTIVE: Practice settings for American psychiatrists were examined for recent trends. METHODS: Surveys were conducted in 1996 (N=970) and 2002 (N=917) among members of the American Psychiatric Association. RESULTS: Between 1996 and 2002 the percentage of direct patient care hours in publicly funded settings increased from 40 to 50 percent for early-career psychiatrists and from 29 to 44 percent for mid-career psychiatrists. By 2002 the percentage of direct patient care hours was higher in publicly funded settings than in solo office practices for early career psychiatrists (50 percent versus 17 percent) and mid-career psychiatrists (44 percent versus 29 percent). CONCLUSIONS: The popular image of the psychiatrist sitting in a private office does not conform with current survey data, which show that psychiatric practice is increasingly taking place in publicly funded settings. Because it extends to mid-career psychiatrists, the shift from private office practice to publicly funded settings is not just a manifestation of early-career psychiatrists' earning a salary while building up their private practices but is a more enduring change in the landscape of psychiatric practice. The authors discuss the implications of these findings with regard to professional identity and training of psychiatrists. PMID- 17085615 TI - Training South African mental health care providers to talk about sex in the era of AIDS. AB - OBJECTIVE: Mental health care providers in South Africa often lack the skills to conduct effective prevention activities in psychiatric settings. This article describes the development and evaluation of an HIV education program for mental health care providers at three psychiatric institutions in South Africa. METHODS: The research team worked with a core group of 16 mental health care providers to assess HIV training needs and to develop a training intervention focused on identified issues. The training intervention was administered to three groups (42 total) during three 1.5-day workshops. Providers completed pre- and post intervention assessments that measured knowledge and attitudes about HIV and AIDS. RESULTS: Data analysis revealed a significant increase in reported levels of comfort with HIV care (d=.54), perceived knowledge of HIV (d=1.17), and factual knowledge (d=.74). CONCLUSIONS: This contextually relevant HIV education curriculum changed providers' attitudes and knowledge, demonstrated the feasibility of administering the training program, and provided a foundation for further prevention activities. PMID- 17085616 TI - Perpetrators of homicide with schizophrenia: a national clinical survey in England and Wales. AB - OBJECTIVE: Few studies have described rates of schizophrenia in a national sample of homicide perpetrators. This study aimed to describe this group's social and clinical characteristics, mental state features, offense details, and outcome in court. METHOD: Analyses used a national clinical survey that collected data on people convicted of homicide in England and Wales (1996-1999). Data were collected for those with schizophrenia or other delusional disorders from psychiatric reports and questionnaires. RESULTS: Of the 1,594 people convicted of homicide, 85 (5 percent) had schizophrenia. Of the 57 people with schizophrenia for whom data were available, 32 (56 percent) had been ill for less than 12 months, and in the month before the offense, 32 (56 percent) had shown a change in the quality, intensity, or conviction of or emotional response to their delusional beliefs. Twenty-four (28 percent) had no previous contact with psychiatric services. CONCLUSIONS: Regular assessment of delusions may help to detect an increased risk of violence, including homicide. More intensive care should be available for patients with a history of schizophrenia and previous violence. PMID- 17085617 TI - A comparison of self-reported use of behavioral health services with Medicaid agency records in Minnesota. AB - OBJECTIVE: This study examined how well recipients of psychiatric or substance abuse services self-report their care. METHODS: Self-reported use of behavioral health services, as reported by 1,012 respondents to a survey mailed to Minnesota Medicaid managed care enrollees receiving services with a behavioral health diagnosis other than severe mental illness in 2000, was compared with Medicaid agency records. RESULTS: Overall, 15 percent of respondents said they had not received services, although administrative data indicated otherwise. In bivariate analyses, failure to report treatment was associated with gender, age, education, and diagnosis but not with race or ethnicity or with residence. The effects of gender and diagnosis were insignificant after analyses controlled for age and education. CONCLUSIONS: Self-reports of behavioral health service use are a relatively accurate method of obtaining information on use. The finding that underreporting of receipt of behavioral health services was not random suggests that reporting errors could introduce bias when comparing groups that differ on age or education. PMID- 17085618 TI - Process versus outcomes in evidence-based practices. PMID- 17085619 TI - Unmasking medical illness in mental health care. PMID- 17085620 TI - SSRI prescription rates after a terrorist attack. PMID- 17085622 TI - Some abnormal psychical conditions in children: excerpts from three lectures. PMID- 17085623 TI - The relevance of the still lectures to attention-deficit/hyperactivity disorder: a commentary. PMID- 17085624 TI - ADHD with comorbid anxiety: a review of the current literature. AB - OBJECTIVE/METHOD: ADHD is often comorbid with anxiety disorders, with rates approaching 25% in many samples. This current review's goal is to examine the literature on ADHD with comorbid anxiety from 1998 to the present. RESULTS: Recent studies indicate that anxiety in ADHD may a) partially inhibit the impulsivity and response inhibition deficits, b) make working memory deficits worse, and c) may be qualitatively different from more phobic types of anxiety seen in pure anxiety samples. In examining subtypes of ADHD, measures of sluggish cognitive tempo show strong correlations with anxiety measures. Insights into the nature of the comorbidity between ADHD and anxiety may be gained by examining the possible comorbidity between Obsessive Compulsive Disorder and ADHD, shared risk factors for ADHD and anxiety, and the current pathogenic models of ADHD. CONCLUSION: The article concludes with a synthesis of the above work, along with directions for future research. PMID- 17085625 TI - A combined treatment approach for adults with ADHD--results of an open study of 43 patients. AB - OBJECTIVE: Increasing numbers of adults are seeking treatment for ADHD. Pharmacotherapy is well established as the first line treatment for adult ADHD, although medications alone may be insufficient treatment for the myriad problems experienced by these patients. Few studies have examined the clinical outcomes of a combination of pharmacotherapy and psychotherapy for this clinical population. The purpose of the present study is to examine the potential effectiveness of the combination of pharmacotherapy and cognitive-behavior therapy modified to treat adult patients diagnosed with ADHD. METHOD: Forty-three adults who underwent a structured diagnostic assessment for ADHD complete 6 months of combined treatment. Treatment outcome is assessed by comparing pre- and posttreatment measures of ADHD symptoms, comorbid symptoms, and ratings of overall functioning. RESULTS/CONCLUSION: Results indicate that combined treatment is associated with significant improvements on all clinical measures. Limitations of the study design and directions for future research are discussed. PMID- 17085626 TI - Does ADHD in adults affect the relative accuracy of metamemory judgments? AB - OBJECTIVE: Prior research suggests that individuals with ADHD overestimate their performance across domains despite performing more poorly in these domains. The authors introduce measures of accuracy from the larger realm of judgment and decision making--namely, relative accuracy and calibration--to the study of self evaluative judgment accuracy in adults with ADHD. METHOD: Twenty-eight adults with ADHD and 28 matched controls participate in a computer-administered paired associate learning task and predict their future recall using immediate and delayed judgments of learning (JOLs). Retrospective confidence judgments are also collected. RESULTS: Groups perform equally in terms of judgment magnitude and absolute judgment accuracy as measured by discrepancy scores and calibration curves. Both groups benefit equally from making their JOL at a delay, and the group with ADHD show higher relative accuracy for delayed judgments. CONCLUSION: Results suggest that under certain circumstances, adults with ADHD can make accurate judgments about their future memory. PMID- 17085627 TI - Magnetic resonance imaging volumetric analysis of the putamen in children with ADHD: combined type versus control. AB - OBJECTIVE: Volumetric differences in the putamen of boys with ADHD combined subtype with psychopathic traits and controls are investigated. METHOD: The putamen in 24 archival magnetic resonance imaging scans of 12 boys in residential treatment with symptoms of ADHD and psychopathic traits and 12 community control boys are analyzed using Display software. RESULTS: There are no differences found in the total, left, and right putamen volumes across the ADHD or control group. A significant reversal of asymmetry across groups is found; children with ADHD more frequently have a smaller left putamen than right. In contrast, the control group more frequently has a smaller right than left putamen. CONCLUSION: A reversal of symmetry in the putamen (as found in the caudate) may relate to ADHD symptomology as well as to psychopathic traits. PMID- 17085628 TI - Children should be seen and heard: self-report of feelings and behaviors in primary-school-age children with ADHD. AB - OBJECTIVE: The belief that children with externalizing disorders have difficulties with self-awareness raises the question of whether children with externalizing disorders are good informants of their own behavior. METHOD: This study investigates how children with ADHD rate their behaviors compared to children without ADHD on a new rating scale (the Self-Evaluation Scale for Children). RESULTS: Preliminary results indicate that this rating scale has acceptable reliability and validity. Furthermore, children with ADHD are found to provide useful information about their feelings and behaviors. Compared to children without ADHD, children with ADHD report more disorganized, disruptive, and impulsive behaviors; poorer self-perception; and poorer social and communication skills. They do not report any less interest in school activities nor more anxiety than the children without ADHD. CONCLUSION: These findings suggest that children with ADHD are more self-aware than previously thought, and this information should inform our clinical and research practice. PMID- 17085629 TI - Effects of coexisting disorders on cognition and behavior in children with ADHD. AB - OBJECTIVE: To determine whether the presence of coexisting disorders affects memory, visual-motor skills, and daily functioning in children with ADHD. METHOD: Eighty-four boys and 18 girls (8 to 16 years old) participate in this study. Twenty children meet criteria for ADHD alone, 42 children for ADHD plus one other disorder, and 40 children for ADHD plus at least two other disorders (reading disability, developmental coordination disorder, oppositional defiant disorder, conduct disorder, anxiety, and depression). Children are assessed on memory and visual-motor skills. Parents complete measures of daily functioning. RESULTS: No significant group differences emerge for age, socioeconomic status, or IQ. Poorer performance on tests of memory and visual-motor skills, more prevalent behavioral problems, and more impairment in everyday functioning are associated with a higher number of coexisting disorders in children with ADHD. CONCLUSION: The presence of coexisting disorders has a significant influence on cognition and behavior of children with ADHD. PMID- 17085630 TI - A preliminary investigation of ADHD symptoms in persons with celiac disease. AB - OBJECTIVE: Several studies report a possible association of celiac disease (CD) with psychiatric and psychological disturbances, such as ADHD. METHOD: The authors assess 132 participants from 3 to 57 years of age (M = 19.3 years) affected by CD for the possibility of an associated ADHD-like symptomatology, using the Conner Scale Hypescheme, a behavioral scale similar to the Conners Rating Scales, before their gluten-free diet was started and 6 months later. RESULTS: The overall score improves significantly as well as most of the ADHD like symptomatology specific features (Bonferroni-corrected, paired-sample t tests). CONCLUSION: The data indicate that ADHD-like symptomatology is markedly overrepresented among untreated CD patients and that a gluten-free diet may improve symptoms significantly within a short period of time. The results of this study also suggest that CD should be included in the list of diseases associated with ADHD-like symptomatology. PMID- 17085631 TI - A controlled study of autonomic nervous system function in adults with attention deficit/hyperactivity disorder treated with stimulant medications: results of a pilot study. AB - OBJECTIVE: Despite the fact that autonomic nervous system (ANS) abnormalities are commonly found in adults and predict increased cardiovascular mortality, no studies have assessed ANS function in adults with attention-deficit/hyperactivity disorder (ADHD) taking stimulants. METHOD: This pilot study evaluated ANS function in adults with ADHD in comparison with an age- and gender-matched control group. RESULTS: The authors found that 4% of the control group had some abnormalities in the ANS in comparison with 24% of the ADHD group. CONCLUSION: Because the control group had higher levels of exercise fitness, and the level of abnormalities in the ADHD group was comparable with that of the general population, the significance of these findings is unknown. In addition, we did not determine if ANS abnormalities were present in individuals with ADHD who were not on stimulant medications. Further research is warranted to determine if there is any association between ADHD and stimulant use and ANS abnormalities. PMID- 17085632 TI - Manual transmission enhances attention and driving performance of ADHD adolescent males: pilot study. AB - OBJECTIVE: Inattention is a major contributor to driving mishaps and is especially problematic among adolescent drivers with ADHD, possibly contributing to their 2 to 4 times higher incidence of collisions. Manual transmission has been demonstrated to be associated with greater arousal. This study tests the hypotheses that manual transmission, compared to automatic transmission, would be associated with better attention and performance on a driving simulator. METHOD: Ten adolescent drivers with ADHD practice driving on the simulator in the manual and automatic mode. Employing a single-blind, cross-over design, participants drive the simulator at 19:30 and 22:30 hr for 30 min in both transmissions and rate their attention to driving. RESULTS: Subjectively, participants report being more attentive while driving in manual transmission mode. Objectively, participants drive safer in the manual transmission mode. CONCLUSION: Although in need of replication, this pilot study suggests a behavioral intervention to improve driving performance among ADHD adolescents. PMID- 17085633 TI - Case series: evaluation of a behavioral sleep intervention for three children with attention-deficit/hyperactivity disorder and dyssomnia. AB - OBJECTIVE: The authors conducted a preliminary evaluation of a behavioral sleep intervention for children with attention-deficit/hyperactivity disorder (ADHD) and dyssomnia delivered via distance treatment. METHOD: Three children (1 male, 2 females; aged 6-10 years) with ADHD and dyssomnia participated in a 5-week manualized intervention. Using a non-concurrent multiple baseline design, the children were randomly assigned to a 1-, 2-, or 3-week baseline and then received the 5-week intervention. Sleep problems were assessed daily by both objective and subjective measures. Measures of problematic nighttime behaviors and daytime ADHD symptoms were also obtained weekly. RESULTS: After 5 weeks, a clinically significant decrease was found in the severity of the children's dyssomnia. No changes in ADHD symptoms were noted. Gains were generally maintained at the 3 month follow-up. Parents perceived the intervention as being "helpful" to "very helpful." CONCLUSION: Based on the initial data, this intervention shows promise as an effective and feasible treatment. PMID- 17085634 TI - Proteomic and microarray analyses of the Dictyostelium Zak1-GSK-3 signaling pathway reveal a role in early development. AB - GskA, the Dictyostelium GSK-3 orthologue, is modified and activated by the dual specificity tyrosine kinase Zak1, and the two kinases form part of a signaling pathway that responds to extracellular cyclic AMP. We identify potential cellular effectors for the two kinases by analyzing the corresponding null mutants. There are proteins and mRNAs that are altered in abundance in only one or the other of the two mutants, indicating that each kinase has some unique functions. However, proteomic and microarray analyses identified a number of proteins and genes, respectively, that are similarly misregulated in both mutant strains. The positive correlation between the array data and the proteomic data is consistent with the Zak1-GskA signaling pathway's functioning by directly or indirectly regulating gene expression. The discoidin 1 genes are positively regulated by the pathway, while the abundance of the H5 protein is negatively regulated. Two of the targets, H5 and discoidin 1, are well-characterized markers for early development, indicating that the Zak1-GskA pathway plays a role in development earlier than previously observed. PMID- 17085635 TI - Carotenoid biosynthesis in the primitive red alga Cyanidioschyzon merolae. AB - Cyanidioschyzon merolae is considered to be one of the most primitive of eukaryotic photosynthetic organisms. To obtain insights into the origin and evolution of the pathway of carotenoid biosynthesis in eukaryotic plants, the carotenoid content of C. merolae was ascertained, genes encoding enzymes of carotenoid biosynthesis in this unicellular red alga were identified, and the activities of two candidate pathway enzymes of particular interest, lycopene cyclase and beta-carotene hydroxylase, were examined. C. merolae contains perhaps the simplest assortment of chlorophylls and carotenoids found in any eukaryotic photosynthetic organism: chlorophyll a, beta-carotene, and zeaxanthin. Carotenoids with epsilon-rings (e.g., lutein), found in many other red algae and in green algae and land plants, were not detected, and the lycopene cyclase of C. merolae quite specifically produced only beta-ringed carotenoids when provided with lycopene as the substrate in Escherichia coli. Lycopene beta-ring cyclases from several bacteria, cyanobacteria, and land plants also proved to be high fidelity enzymes, whereas the structurally related epsilon-ring cyclases from several plant species were found to be less specific, yielding products with beta rings as well as epsilon-rings. C. merolae lacks orthologs of genes that encode the two types of beta-carotene hydroxylase found in land plants, one a nonheme diiron oxygenase and the other a cytochrome P450. A C. merolae chloroplast gene specifies a polypeptide similar to members of a third class of beta-carotene hydroxylases, common in cyanobacteria, but this gene did not produce an active enzyme when expressed in E. coli. The identity of the C. merolae beta-carotene hydroxylase therefore remains uncertain. PMID- 17085636 TI - Deletion of the glucose-6-phosphate dehydrogenase gene KlZWF1 affects both fermentative and respiratory metabolism in Kluyveromyces lactis. AB - In Kluyveromyces lactis, the pentose phosphate pathway is an alternative route for the dissimilation of glucose. The first enzyme of the pathway is the glucose 6-phosphate dehydrogenase (G6PDH), encoded by KlZWF1. We isolated this gene and examined its role. Like ZWF1 of Saccharomyces cerevisiae, KlZWF1 was constitutively expressed, and its deletion led to increased sensitivity to hydrogen peroxide on glucose, but unlike the case for S. cerevisiae, the Klzwf1Delta strain had a reduced biomass yield on fermentative carbon sources as well as on lactate and glycerol. In addition, the reduced yield on glucose was associated with low ethanol production and decreased oxygen consumption, indicating that this gene is required for both fermentation and respiration. On ethanol, however, the mutant showed an increased biomass yield. Moreover, on this substrate, wild-type cells showed an additional band of activity that might correspond to a dimeric form of G6PDH. The partial dimerization of the G6PDH tetramer on ethanol suggested the production of an NADPH excess that was negative for biomass yield. PMID- 17085637 TI - alpha-tubulin minichromosome promoters in the stichotrichous ciliate Stylonychia lemnae. AB - Ciliated protists are model organisms for a number of molecular phenomena including telomerase function, self-splicing introns, and an RNA interference related mechanism in programmed DNA elimination. Despite this relevance, our knowledge about promoters and transcriptional regulation in these organisms is very limited. The macronuclear genome of stichotrichous ciliates consists of minichromosomes which typically encode a single gene. The 5' nontranscribed spacers are usually no longer than 400 bp and highly suitable for promoter characterizations. We used microinjection of two artificial and differently tagged alpha1 tubulin minichromosomes into the macronucleus of Stylonychia lemnae as a means to characterize in detail the corresponding promoter. Clonal cell lines that stably maintained both minichromosomes were generated, enabling comparative expression analysis by primer extension assays. Deletion and block substitution mutations of one of the minichromosomes revealed a TATA-like element, a putative initiator element, and two distinct upstream sequence elements (USEs). Determination of transcription initiation sites and a sequence alignment indicated that both TATA-like and initiator elements are conserved components of S. lemnae minichromosomes, whereas the USEs appear to be specific for the alpha1 tubulin minichromosome. The alpha2 tubulin minichromosome promoter is very short, comprising the two proximal elements but not the USEs. Despite the latter finding, up-regulation of alpha-tubulin expression in cells treated with concanavalin A activated the alpha2 but not the alpha1 tubulin promoter. These results therefore show that gene expression regulation in S. lemnae occurs at the level of transcription initiation on the basis of structurally different promoters. PMID- 17085638 TI - Toxoplasma gondii targets a protein phosphatase 2C to the nuclei of infected host cells. AB - Intracellular pathogens have evolved a wide array of mechanisms to invade and co opt their host cells for intracellular survival. Apicomplexan parasites such as Toxoplasma gondii employ the action of unique secretory organelles named rhoptries for internalization of the parasite and formation of a specialized niche within the host cell. We demonstrate that Toxoplasma gondii also uses secretion from the rhoptries during invasion to deliver a parasite-derived protein phosphatase 2C (PP2C-hn) into the host cell and direct it to the host nucleus. Delivery to the host nucleus does not require completion of invasion, as evidenced by the fact that parasites blocked in the initial stages of invasion with cytochalasin D are able to target PP2C-hn to the host nucleus. We have disrupted the gene encoding PP2C-hn and shown that PP2C-hn-knockout parasites exhibit a mild growth defect that can be rescued by complementation with the wild type gene. The delivery of parasite effector proteins via the rhoptries provides a novel mechanism for Toxoplasma to directly access the command center of its host cell during infection by the parasite. PMID- 17085639 TI - Circulating tumor cells in breast cancer: blood will tell. PMID- 17085640 TI - Is the type I insulin-like growth factor receptor a therapeutic target in endometrial cancer? PMID- 17085641 TI - Advances in targeting human epidermal growth factor receptor-2 signaling for cancer therapy. AB - Human epidermal growth factor receptor (HER)-2 is a member of the HER tyrosine kinase family, which regulates cell growth and proliferation. HER-2 is overexpressed in 20% to 30% of breast cancers and has been associated with an aggressive phenotype and a poorer prognosis, making it an appealing therapeutic target. Since 1998, the anti-HER-2 antibody trastuzumab has been used for the treatment of women with HER-2-positive metastatic breast cancer. Results from large trials have established a role for trastuzumab in the adjuvant setting for the treatment of high-risk primary breast cancer as well. Tyrosine kinase inhibitors that target HER-2 are also very promising therapies and are likely to be incorporated into clinical practice in the near future. HER-2-targeted therapies represent a major step forward in achieving our goal of delivering individualized targeted therapy for breast cancer. However, there are many unanswered questions about the optimal use of these agents. Ongoing research will better elucidate the best combination therapies to overcome resistance to HER-2 targeted agents and will help identify patients at high enough risk to warrant their toxicity. PMID- 17085642 TI - Attractin is elevated in the cerebrospinal fluid of patients with malignant astrocytoma and mediates glioma cell migration. AB - PURPOSE: There are a limited number of noninvasive methods available for the monitoring of neoplastic disease in the central nervous system. The goal of our study was to find reliable markers that could be used for disease monitoring as well as to identify new targets for the therapeutic intervention for malignant astrocytoma (WHO grades 3 and 4). EXPERIMENTAL DESIGN: We employed proteomic techniques to identify secreted proteins in the cerebrospinal fluid that were specific to patients with malignant astrocytoma. RESULTS: Among 60 cerebrospinal fluid samples of patients with various central nervous system diseases, attractin was consistently found to be elevated in the samples of patients with malignant astrocytoma. To independently validate these results, we examined attractin expression in a new set of 108 normal and tumoral brain tissue specimens and found elevated expression in 97% of malignant astrocytomas, with the highest levels in grade 4 tumors. Using immunohistochemistry, we further showed that attractin is produced and secreted by the tumor cells. Finally, we showed that cerebrospinal fluid from brain tumor patients induces glioma cell migration and that attractin is largely responsible for this promigratory activity. CONCLUSIONS: Our results find attractin to be a reliable secreted marker for high grade gliomas. Additionally, our migration studies suggest that it may be an important mediator of tumor invasiveness, and thus, a potential target in future therapies. PMID- 17085643 TI - Metastasis-suppressing potential of ribonucleotide reductase small subunit p53R2 in human cancer cells. AB - PURPOSE: Previous gene transfection studies have shown that the accumulation of human ribonucleotide reductase small subunit M2 (hRRM2) enhances cellular transformation, tumorigenesis, and malignancy potential. The latest identified small subunit p53R2 has 80% homology to hRRM2. Here, we investigate the role of p53R2 in cancer invasion and metastasis. EXPERIMENTAL DESIGN: The immunohistochemistry was conducted on a tissue array including 49 primary and 59 metastatic colon adenocarcinoma samples to determine the relationship between p53R2 expression and metastasis. A Matrigel invasive chamber was used to sort the highly invasive cells and to evaluate the invasion potential of p53R2. RESULTS: Univariate and multivariate analyses revealed that p53R2 is negatively related to the metastasis of colon adenocarcinoma samples (odds ratio, 0.23; P<0.05). The decrease of p53R2 is associated with cell invasion potential, which was observed in both p53 wild-type (KB) and mutant (PC-3 and Mia PaCa-2) cell lines. An increase in p53R2 expression by gene transfection significantly reduced the cellular invasion potential to 54% and 30% in KB and PC-3 cells, respectively, whereas inhibition of p53R2 by short interfering RNA resulted in a 3-fold increase in cell migration. CONCLUSIONS: Opposite regulation of hRRM2 and p53R2 in invasion potential might play a critical role in determining the invasion and metastasis phenotype in cancer cells. The expression level of ribonucleotide reductase small subunits may serve as a biomarker to predict the malignancy potential of human cancers in the future. PMID- 17085644 TI - Acceleration of telomere loss by chemotherapy is greater in older patients with locally advanced head and neck cancer. AB - PURPOSE: Chronic viral infection and combinations of chemotherapeutic drugs have been reported to accelerate telomere erosion. Here, we asked if chemoradiotherapy, using the single agent cisplatin, would accelerate telomere loss in head and neck cancer patients, and whether loss was linked to smoking status, age, gender, or stage of disease at diagnosis. EXPERIMENTAL DESIGN: Blood samples were collected from 20 patients with squamous cell cancer of the head and neck before, during, and after chemoradiotherapy. Following DNA isolation from peripheral blood mononuclear cells, telomere length was measured by terminal restriction fragment analysis. RESULTS: Chemoradiotherapy increased the rate of telomere erosion>100-fold. Telomere length before treatment in chemoradiotherapy patients was similar to age-matched controls. Although smokers began with significantly shorter telomeres, smoking status did not affect chemoradiotherapy induced attrition, nor did gender or stage of disease. We also make the novel observation that a significantly greater telomere loss occurred in response to treatment in older patients, with those younger than 55 years losing an average of 400 bp of telomeric DNA compared with the 880 bp lost by those over 55 years. CONCLUSIONS: The lack of telomere length difference before treatment suggests that shortened telomeres may not be a risk factor for development of head and neck cancer in the age range we examined. Chemoradiotherapy caused a severe telomere length reduction in all patients. The significant difference seen in the elderly (P=0.018) suggests that chemoradiotherapy may have more severe effects on the replicative capacity of blood cells in older patients. PMID- 17085645 TI - Activation of hypoxia-inducible factor-1alpha is necessary for lysophosphatidic acid-induced vascular endothelial growth factor expression. AB - PURPOSE: Lysophosphatidic acid (LPA) plays an important role in mediating cell proliferation, survival, and tumor invasion and angiogenesis. This bioactive phospholipid at the concentration in ascitic fluid stimulates the growth of malignant ovarian tumors by increasing the expression of vascular endothelial growth factor (VEGF). In the present study, we investigated whether LPA activates hypoxia inducible factor-1 (HIF-1), a key transcriptional complex in tumor progression and metastasis, thereby increasing the expression of VEGF. EXPERIMENTAL DESIGN: Immunoblotting, reverse transcription-PCR, ELISA, immunofluorescence, and chromatin immunoprecipitation assay were used to examine the expression of VEGF and HIF-1alpha in various cancer cells. Specific HIF 1alpha small interfering RNA was transfected to various cancer cells to determine the role of HIF-1alpha in LPA-induced VEGF expression. RESULTS: LPA induced expressions of VEGF and HIF-1alpha in OVCAR-3, CAOV-3, PC-3, and SK-Hep1 cells but not in SKOV-3 and Hep-3B cells. In OVCAR-3 and PC-3 cells, the phosphoinositide 3-kinase/Akt/mammalian target of rapamycin/p70S6K and p42/p44 mitogen-activated protein kinase pathways were required for LPA-induced HIF 1alpha and VEGF expressions, whereas only the phosphoinositide 3-kinase/mammalian target of rapamycin/p70S6K pathway was important in SK-Hep1 cells. Immunofluorescence microscopy assay showed translocation of HIF-1alpha to nucleus by LPA, and chromatin immunoprecipitation assay revealed the binding of HIF 1alpha to the promoter of VEGF by LPA. Importantly, we found that small interfering RNA-induced reduction of HIF-1alpha expression significantly attenuated VEGF expression by LPA. CONCLUSIONS: Our results show for the first time that LPA induces VEGF via HIF-1alpha activation and reveal a critical role of HIF-1alpha in LPA-induced cancer cell proliferation and angiogenesis. PMID- 17085646 TI - Distribution of GPR30, a seven membrane-spanning estrogen receptor, in primary breast cancer and its association with clinicopathologic determinants of tumor progression. AB - PURPOSE: The seven transmembrane receptor, GPR30, is linked to estrogen binding and heparan-bound epidermal growth factor release. Here, the significance of GPR30 in human breast cancer was evaluated by comparing its relationship to steroid hormone receptor expression and tumor progression variables. EXPERIMENTAL DESIGN: Immunohistochemical analysis of a National Cancer Institute-sponsored tumor collection comprised of 361 breast carcinomas obtained at first diagnosis (321 invasive and 40 intraductal tumors). Biopsies from 12 reduction mammoplasties served as controls. The distribution pattern of GPR30, estrogen receptor (ER), and progesterone receptor (PR) was correlated with clinicopathologic variables obtained at diagnosis. RESULTS: GPR30, ER, and PR were positive in all 12 normal controls. In contrast, GPR30 expression varied in breast tumors, in which 62% (199 of 321) of invasive tumors and 42% (17 of 40) of intraductal tumors were positive. Codistribution of ER and GPR30 was measured in 43% (139 of 321) of invasive breast tumors, whereas both receptors were lacking (ER-GPR30-) in 19% (61 of 321) of the tumors analyzed, indicating a significant association between ER and GPR30 (P<0.05). The coexpression of PR and ER did not influence GPR30 expression, yet coexpression of GPR30 and ER was linked to PR positivity. Unlike ER, which varied inversely with HER-2/neu and tumor size, GPR30 positively associated with HER-2/neu and tumor size. In addition, GPR30 showed a positive association with metastasis (P=0.014; odds ratio, 1.9). CONCLUSIONS: GPR30 and ER exhibited distinct patterns of association with breast tumor progression variables, including HER-2/neu, tumor size, and metastatic disease. Thus, these results support the hypothesis that GPR30 and ER have an independent influence on estrogen responsiveness in breast carcinoma. PMID- 17085647 TI - Identification of the decay-accelerating factor CD55 as a peanut agglutinin binding protein and its alteration in non-small cell lung cancers. AB - PURPOSE: Peanut agglutinin (PNA) recognizes tumor-associated carbohydrates. In this study, we aimed to identify the core protein harboring PNA-binding sugars in the human lung and to explore the relationship with the pathology of primary non small cell lung cancers (NSCLC). EXPERIMENTAL DESIGN: PNA lectin blotting was used to detect PNA-binding proteins in the microsomal fraction of lung tissue from 24 patients with NSCLC. The 55- to 65-kDa core peptide PNA-binding protein was characterized by enzymatic treatment and identified by immunoprecipitation and affinity chromatography. The expression level and increase in size of the 55- to 65-kDa PNA-binding protein/decay-accelerating factor (DAF) were compared between normal and tumor regions of the tumor tissue by Western blotting and quantitative PCR. RESULTS: The 55- to 65-kDa PNA-binding protein was observed in human lung. This was a glycosylphosphatidylinositol-anchored membrane protein carrying O-linked carbohydrates. This core protein was identified as DAF, one of the complementary regulatory proteins. DAF was enlarged to 65 to 75 kDa in NSCLC tumor lesions due to sialylation in the sugar moiety. At the transcription level, DAF levels were significantly lower in tumor regions, suggesting its down regulation in NSCLC cells. CONCLUSIONS: DAF was identified as a new PNA-binding protein in the human lung. The down-regulation and heavy sialylation of DAF was associated with pathology in NSCLC, and these alterations make this protein a potential marker for NSCLC. PMID- 17085648 TI - Overexpression of the insulin-like growth factor I receptor and activation of the AKT pathway in hyperplastic endometrium. AB - PURPOSE: Although there is considerable information on the molecular aberrations associated with endometrial cancer, very little is known of the changes in gene expression associated with endometrial hyperplasia. EXPERIMENTAL DESIGN: To address this, we have compared the level of expression of estrogen-regulated genes and components of the insulin-like growth factor I (IGF-I) signaling pathway in endometrial biopsies from subjects with normal endometrium, complex atypical endometrial hyperplasia, and endometrial adenocarcinoma (type I). RESULTS: There was a significant increase in the expression of the IGF-I receptor (IGF-IR) in biopsies from hyperplastic endometrium and endometrial carcinoma compared with the proliferative endometrium. The receptor was also activated, as judged by increased tyrosine phosphorylation. In addition, in endometrial hyperplasia and carcinoma, the downstream components of the IGF-IR pathway are activated, as reflected in increased Akt phosphorylation. Loss of phosphatase and tensin homologue deleted on chromosome 10 (PTEN) expression in endometrial hyperplasia did not correlate with increased activation of IGF-IR. However, the simultaneous loss of PTEN expression and increased IGF-IR activation in hyperplasia was associated with an increased incidence of endometrial carcinoma. CONCLUSIONS: These results suggest that up-regulation of IGF-IR and loss of PTEN may be independent events that give rise to complementary activation of the IGF-I pathway and increase the probability of the development of cancer. These studies suggest that increased expression of IGF-IR may be an important contributor to the risk of endometrial hyperplasia and cancer. PMID- 17085649 TI - Chromosomal instability in microsatellite-unstable and stable colon cancer. AB - PURPOSE: The genomic instability in colon cancer can be divided into at least two major types, microsatellite instability (MSI) or chromosomal instability (CIN). Although initially felt to be mutually exclusive, recent evidence suggests that there may be overlap between the two. The aim of this study was to identify chromosomal alterations at high resolution in sporadic colon cancers with high level microsatellite instability (MSI-H) and to compare them to those present in a set of matched microsatellite stable (MSS) tumors. EXPERIMENTAL DESIGN: Array based comparative genomic hybridization was used to analyze a set of 23 sporadic MSI-H and 23 MSS colon cancers matched for location, gender, stage, and age. The arrays consisted of 2,464 bacterial artificial chromosome clones. RESULTS: MSI and MSS colon cancers differed significantly with respect to frequency and type of chromosomal alterations. The median fraction of genome altered was lower among MSI-H tumors than MSS tumors (2.8% versus 30.7%, P=0.00006). However, the MSI-H tumors displayed a range of genomic alterations, from the absence of detectable alterations to extensive alterations. Frequent alterations in MSI-H tumors included gains of chromosomes 8, 12, and 13, and loss of 15q14. In contrast, the most frequent alterations in MSS tumors were gains of 7, 13, 8q, and 20, and losses of 8p, 17p, and 18. A small, previously uncharacterized, genomic deletion on 16p13.2, found in 35% of MSI-H and 21% of MSS tumors, was confirmed by fluorescence in situ hybridization. CONCLUSION: MSI and CIN are not mutually exclusive forms of genomic instability in sporadic colon cancer, with MSI tumors also showing varying degrees of CIN. PMID- 17085650 TI - RUNX3 inhibits the expression of vascular endothelial growth factor and reduces the angiogenesis, growth, and metastasis of human gastric cancer. AB - PURPOSE: Recent studies indicated that RUNX3 exhibits potent antitumor activity. However, the underlying molecular mechanisms of this activity remain unclear. In the present study, we used a gastric cancer model to determine the effect of RUNX3 expression on tumor angiogenesis. EXPERIMENTAL DESIGN: The effects of increased RUNX3 expression on vascular endothelial growth factor (VEGF) expression in and angiogenic potential of human gastric cancer cells were determined in vitro and in animal models. RUNX3 and VEGF expression was determined in 120 human gastric cancer specimens and their relationship was analyzed. RESULTS: RUNX3 gene transfer suppressed VEGF expression in human gastric cancer cells. Down-regulation of VEGF expression correlated with a significantly impaired angiogenic potential of human gastric cancer cells. Furthermore, RUNX3 restoration inhibited tumor growth and metastasis in animal models, which was consistent with inhibition of angiogenesis as determined by evaluating VEGF expression and tumor microvessel formation. In gastric cancer specimens, loss or decrease in RUNX3 expression inversely associated with increased VEGF expression and elevated microvessel formation. CONCLUSIONS: Our clinical and experimental data provide a novel molecular mechanism for the antitumor activity of RUNX3 and may help design effective therapy targeting RUNX3 pathway to control gastric cancer growth and metastasis. PMID- 17085651 TI - Loss of Kruppel-like factor 4 expression contributes to Sp1 overexpression and human gastric cancer development and progression. AB - PURPOSE: Increasing evidence indicates that the transcription factor, Sp1, regulates the expression of multiple genes involved in tumor development and progression. We have recently reported that Sp1 overexpression is directly correlated with the angiogenic potential of and poor prognosis for human gastric cancer. However, the underlying mechanisms that result in Sp1 overexpression remain unclear. EXPERIMENTAL DESIGN: The expression of Sp1 and Kruppel-like factor 4 (KLF4), a potential tumor suppressor gene, in gastric cancer tissue was analyzed by immunohistochemistry and Western blot analysis. Alterations of Sp1 and KLF4 expression were achieved by gene transfer and verified by Northern and Western blot analyses. Furthermore, Sp1 promoter activity assay, electrophoretic mobility shift assay, and chromatin immunoprecipitation assay were done to identify the KLF4 binding sites on the Sp1 promoter. RESULTS: Mutually exclusive expression of Sp1 and KLF4 was evident in gastric cancer and noncancerous tissue. Specifically, strong Sp1 expression but loss of KLF4 expression was found in cancer tissue, whereas the adjacent noncancerous tissue showed negative Sp1 expression but strong KLF4 expression. Enforced KLF4 expression repressed Sp1 expression at the promoter activity, mRNA, and protein levels. Moreover, a region within the proximal Sp1 promoter was identified to have overlapping KLF4- and Sp1 binding sites, to which KLF4 and Sp1 compete for binding. Sp1 positively regulated its own promoter, whereas KLF4 did the opposite. CONCLUSIONS: Our data suggests that disruption of KLF4-mediated negative regulation contributes to the molecular events of Sp1 overexpression and to the development and progression of human gastric cancer. PMID- 17085652 TI - Circulating tumor cells versus imaging--predicting overall survival in metastatic breast cancer. AB - PURPOSE: The presence of >or=5 circulating tumor cells (CTC) in 7.5 mL blood from patients with measurable metastatic breast cancer before and/or after initiation of therapy is associated with shorter progression-free and overall survival. In this report, we compared the use of CTCs to radiology for prediction of overall survival. EXPERIMENTAL DESIGN: One hundred thirty-eight metastatic breast cancer patients had imaging studies done before and a median of 10 weeks after the initiation of therapy. All scans were centrally reviewed by two independent radiologists using WHO criteria to determine radiologic response. CTC counts were determined approximately 4 weeks after initiation of therapy. Specimens were analyzed at one of seven laboratories and reviewed by a central laboratory. RESULTS: Interreader variability for radiologic responses and CTC counts were 15.2% and 0.7%, respectively. The median overall survival of 13 (9%) patients with radiologic nonprogression and >or=5 CTCs was significantly shorter than that of the 83 (60%) patients with radiologic nonprogression and <5 CTCs (15.3 versus 26.9 months; P=0.0389). The median overall survival of the 20 (14%) patients with radiologic progression and <5 CTCs was significantly longer than the 22 (16%) patients with >or=5 CTCs that showed progression by radiology (19.9 versus 6.4 months; P=0.0039). CONCLUSIONS: Assessment of CTCs is an earlier, more reproducible indication of disease status than current imaging methods. CTCs may be a superior surrogate end point, as they are highly reproducible and correlate better with overall survival than do changes determined by traditional radiology. PMID- 17085653 TI - Reduced expression of the breast cancer metastasis suppressor 1 mRNA is correlated with poor progress in breast cancer. AB - PURPOSE: It is well established that breast cancer metastasis suppressor 1 (BRMS1) suppresses metastasis of breast cancer in animal models without affecting the growth of the primary tumor. It has also been shown to suppress the metastasis of tumors derived from breast and melanoma cell lines. However, its clinical importance for breast cancer patients remains undetermined. This prompted us to investigate its expression level in breast cancer patients to clarify its clinical significance. EXPERIMENTAL DESIGN: The level of expression of BRMS1 mRNA was assessed by LightCycler quantitative real-time reverse transcription-PCR in 161 cases of invasive carcinoma of the breast. Associations between BRMS1 mRNA expression and various clinicopathologic factors were sought. RESULTS: It was found that BRMS1 mRNA was expressed at significantly higher levels in patients>50 years of age, with tumor size<2 cm, or with progesterone receptor-positive and HER2-negative tumors. No differences were found between BRMS1 mRNA expression and axillary lymph node metastasis and histologic grade groups. Patients with high levels of expression of BRMS1 mRNA have a better prognosis than those with low expression. Univariate and multivariate prognostic analysis showed that BRMS1 mRNA is an independent prognostic factor for disease free survival in breast cancer. CONCLUSIONS: These results provide clinical evidence to support the notion that BRMS1 is a breast carcinoma metastasis suppressor gene. Our results also suggest that measuring BRMS1 expression will help to identify those breast cancer patients with worse disease-free survival. PMID- 17085654 TI - Proteomics-based approach identifying autoantibody against peroxiredoxin VI as a novel serum marker in esophageal squamous cell carcinoma. AB - PURPOSE: Detection of novel tumor-related antigens and autoantibodies will aid in diagnosis of early-stage cancer and in development of more effective immunotherapies. The purpose of this study was to identify novel tumor antigens in an esophageal squamous cell carcinoma (ESCC) cell line (TE-2) and related autoantibodies in sera from patients with ESCC using a proteomics-based approach. EXPERIMENTAL DESIGN: TE-2 proteins were separated by two-dimensional PAGE, followed by Western blot analysis in which sera of patients with ESCC, healthy controls, and patients with other cancers were tested for primary antibodies. Positive spots were excised from silver-stained gels and analyzed by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF/TOF MS). RESULTS: Sera from patients with ESCC yielded multiple spots, one of which was identified as peroxiredoxin (Prx) VI by MALDI-TOF/TOF MS. Western blot analysis against recombinant Prx VI showed reactivity in sera from 15 of 30 (50%) patients with ESCC and 2 of 30 (6.6%) healthy individuals. Autoantibody against Prx VI was found in sera from 1 of 30 (3.3%) patients with other types of cancer (colon cancer). CONCLUSION: We have identified for the first time an autoantibody against Prx VI in ESCC patients. The proteomic approach implemented here offers a powerful tool for identifying novel serum markers that may display clinical usefulness against cancer. PMID- 17085655 TI - CA IX is an independent prognostic marker in premenopausal breast cancer patients with one to three positive lymph nodes and a putative marker of radiation resistance. AB - PURPOSE: Hypoxia in breast cancer is associated with poor prognosis and down regulation of the estrogen receptor. Carbonic anhydrase IX (CA IX) is a hypoxia inducible gene that has been associated with poor outcome in many epithelial cancers. Previous studies of CA IX in breast cancer have been carried out on mixed cohorts of premenopausal and postmenopausal patients with locally advanced disease and varying treatment regimens. We examined the potential prognostic and predictive role of CA IX in premenopausal breast cancer patients. EXPERIMENTAL DESIGN: Using tissue microarrays, we analyzed CA IX expression in 400 stage II breast cancers from premenopausal women. The patients had previously participated in a randomized control trial comparing 2 years of tamoxifen to no systemic adjuvant treatment. Median follow-up was 13.9 years. RESULTS: CA IX expression correlated positively with tumor size, grade, hypoxia-inducible factor 1alpha, Ki 67, cyclin E, and cyclin A2 expression. CA IX expression correlated negatively with cyclin D1, estrogen receptor, and progesterone receptor. CA IX expression was associated with a reduced relapse-free survival (P=0.032), overall survival (P=0.022), and breast cancer-specific survival (P=0.005). Multivariate analysis revealed that CA IX was an independent prognostic marker in untreated patients with one to three positive lymph nodes (hazard ratio, 3.2; 95% confidence interval, 1.15-9.13; P=0.027). CONCLUSION: CA IX is marker of poor prognosis in premenopausal breast cancer patients and it is an independent predictor of survival in patients with one to three positive lymph nodes. As all these patients received locoregional radiation therapy, CA IX may be associated with resistance to radiotherapy. PMID- 17085656 TI - Prognostic value of apoptosis in rectal cancer patients of the dutch total mesorectal excision trial: radiotherapy is redundant in intrinsically high apoptotic tumors. AB - PURPOSE: The combination of radiotherapy and good quality surgery reduces local recurrence rate for rectal cancer patients. This study assesses the prognostic value of both intrinsic and radiotherapy-induced apoptosis and evaluates the relevance of radiotherapy for outcome of rectal cancer patients. EXPERIMENTAL DESIGN: Tumor samples (1,198) were available from the Dutch Total Mesorectal Excision trial, in which rectal cancer patients were treated with standardized surgery and randomized for preoperative short-term radiotherapy or not. Tumor samples were obtained at time of surgery. Tissue microarrays were constructed and stained with the active caspase-specific M30 antibody to determine the amount of apoptotic epithelial tumor cells. RESULTS: Nonirradiated patients with a negative circumferential margin displaying lower than median levels of apoptosis developed more local recurrences (10.5% versus 6.1%; P=0.06) and more rapidly after surgery than patients with high intrinsic apoptosis in their tumors (median time to recurrence, 13.0 versus 21.3 months; P=0.04). In multivariate analysis, intrinsic apoptosis was an independent predictor for the development of local recurrences (hazard ratio, 2.0; P=0.05). Radiotherapy increased apoptosis level (11 versus 23 apoptotic cells/mm2 tumor epithelium; P<0.001), but this apoptosis did not influence patients' prognosis. CONCLUSIONS: Rectal cancer patients with low intrinsic apoptosis will benefit from radiotherapy with respect to the development of local recurrences. Because apoptosis is an inherent characteristic of tumors, patients who do not need radiotherapy may be selected based on the apoptotic index of the primary tumor. PMID- 17085657 TI - Sequential positron emission tomography using [18F]fluorodeoxyglucose for monitoring response to chemotherapy in metastatic breast cancer. AB - PURPOSE: To evaluate the clinical value of positron emission tomography (PET) for monitoring chemotherapy in metastatic breast cancer. EXPERIMENTAL DESIGN: Twenty patients with hormonorefractory or hormonoreceptor-negative multimetastatic breast cancer were prospectively included. PET studies were done at baseline, at day 21 after the first cycle and at day 21 after the third cycle of chemotherapy. Metabolic response was defined based on visual and various modes of standardized uptake value (SUV) analysis of sequential PET studies. RESULTS: After one cycle, PET indicated a partial response in 12 patients, stable disease in 7 patients, and progressive disease in 1 patient, according to the visual analysis. After three cycles, PET showed a complete response in 5 patients, partial response in 11 patients, stable disease in 3 patients, and progressive disease in 1 patient. Seventy-five percent of the patients showing a metabolic response on visual analysis effectively responded to the treatment. The average SUV decreased on both the second and the third PET study, but only changes measured after three cycles of chemotherapy predicted the clinical response to chemotherapy and the overall survival. All methods for calculating the SUV (normalized for body weight, body surface area, or lean body mass) provided similar results. CONCLUSION: Semiquantitative analysis of [18F]fluorodeoxyglucose-PET studies done after three cycles of chemotherapy is useful for monitoring the response to chemotherapy in metastatic breast cancer. PMID- 17085658 TI - Peroxisome proliferator-activated receptor gamma is highly expressed in pancreatic cancer and is associated with shorter overall survival times. AB - PURPOSE: Peroxisome proliferator-activated receptor gamma (PPARgamma) is a ligand activated transcription factor that has been implicated in carcinogenesis and progression of various solid tumors, including pancreatic carcinoma. We aimed to clarify the expression patterns of PPARgamma in pancreatic ductal carcinomas and to correlate these to clinicopathologic variables, including patient survival. EXPERIMENTAL DESIGN: Array-based expression profiling of 19 microdissected carcinomas and 14 normal ductal epithelia was conducted. Additionally, Western blots of pancreatic cancer cell lines and paraffinized tissue of 129 pancreatic carcinomas were immunostained for PPARgamma. For statistical analysis, Fisher's exact test, chi2 test for trends, correlation analysis, Kaplan-Meier analysis, and Cox's regression were applied. RESULTS: Expression profiles showed a strong overexpression of PPARgamma mRNA (change fold, 6.9; P=0.04). Immunohistochemically, PPARgamma expression was seen in 71.3% of pancreatic cancer cases. PPARgamma expression correlated positively to higher pT stages and higher tumor grade. Survival analysis showed a significant prognostic value for PPARgamma, which was found to be independent in the clinically important subgroup of node-negative tumors. CONCLUSIONS: PPARgamma is commonly up-regulated in pancreatic ductal adenocarcinoma and might be a prognostic marker in this disease. Both findings corroborate the importance of PPARgamma in tumor progression of pancreatic cancer. PMID- 17085659 TI - Variants of the hK2 protein gene (KLK2) are associated with serum hK2 levels and predict the presence of prostate cancer at biopsy. AB - PURPOSE: Increased levels of serum human kallikrein-2 (hK2) and an hK2 gene (KLK2) variant are positively associated for prostate cancer, but the relationships between them remain unclear. We examined five variants of the KLK2 gene to further define its relevance to prostate cancer susceptibility and hK2 levels. EXPERIMENTAL DESIGN: We genotyped 645 men with biopsy-proven prostate cancer (cases) and 606 males with biopsies negative for prostate cancer (controls) for five additional single nucleotide polymorphisms (SNP) across the KLK2 gene and also tested for serum hK2 levels. These SNPs were identified from sequencing the KLK2 gene among 20 patients with aggressive prostate cancer. Odds ratios (OR) for prostate cancer detection and haplotype analysis were done. RESULTS: Among the SNPs studied, the A allele of the KLK2-SNP1 (G>A, rs2664155) and the T allele of the KLK2-SNP5 (C>T, rs198977) polymorphisms showed positive associations with prostate cancer, adjusted ORs for KLK2-SNP1 AG and AA genotypes being 1.4 [95% confidence interval (95% CI), 1.2-1.8; P=0.002] and for KLK2-SNP5 TT or CT genotypes being 1.3 (95% CI, 1.1-1.6; P=0.05). Haplotype analyses also revealed a significant association between prostate cancer and the haplotype containing both risk alleles (ACCTT), OR being 5.1 (95% CI, 1.6-6.5; P=0.005). Analysis of serum hK2 revealed hK2 levels to be significantly increased in association with KLK2-SNP1 AA and AG risk genotypes compared with the GG genotype (P=0.001) and also in association with the ACCTT risk haplotype compared with the most common non-risk haplotype (P=0.05). CONCLUSIONS: These findings suggest a role for the KLK2 gene in prostate cancer susceptibility and imply that this role may be realized at least in part by the induction of increases in hK2 production. PMID- 17085660 TI - Classification of breast cancer using genetic algorithms and tissue microarrays. AB - PURPOSE: A multitude of breast cancer mRNA profiling studies has stratified breast cancer and defined gene sets that correlate with outcome. However, the number of genes used to predict patient outcome or define tumor subtypes by RNA expression studies is variable, nonoverlapping, and generally requires specialized technologies that are beyond those used in the routine pathology laboratory. It would be ideal if the familiarity and streamlined nature of immunohistochemistry could be combined with the rigorously quantitative and highly specific properties of nucleic acid-based analysis to predict patient outcome. EXPERIMENTAL DESIGN: We have used AQUA-based objective quantitative analysis of tissue microarrays toward the goal of discovery of a minimal number of markers with maximal prognostic or predictive value that can be applied to the conventional formalin-fixed, paraffin-embedded tissue section. RESULTS: The minimal discovered multiplexed set of tissue biomarkers was GATA3, NAT1, and estrogen receptor. Genetic algorithms were then applied after division of our cohort into a training set of 223 breast cancer patients to discover a prospectively applicable solution that can define a subset of patients with 5 year survival of 96%. This algorithm was then validated on an internal validation set (n=223, 5-year survival=95.8%) and further validated on an independent cohort from Sweden, which showed 5-year survival of 92.7% (n=149). CONCLUSIONS: With further validation, this test has both the familiarity and specificity for widespread use in management of breast cancer. More generally, this work illustrates the potential for multiplexed biomarker discovery on the tissue microarray platform. PMID- 17085661 TI - Gastric and intestinal phenotypic marker expression in early differentiated-type tumors of the stomach: clinicopathologic significance and genetic background. AB - PURPOSE: Gastric and intestinal phenotypic cell markers are expressed in gastric carcinomas, irrespective of their histologic type. In the present study, we determined the clinicopathologic significance of phenotypic marker expression in early-stage gastric differentiated-type tumors and the association between marker expression and genetic alterations. EXPERIMENTAL DESIGN: Phenotypic marker expression was determined by examining the expressions of human gastric mucin (HGM), MUC6, MUC2, and CD10 in 63 gastric adenomas, 133 early differentiated-type carcinomas, and 24 follow-up cases with gastric adenoma. Tumors were classified into gastric, gastric and intestinal mixed, or intestinal phenotypes according to the immunopositivity of the above markers. The presence of mutations in APC, K ras, and p53 and the microsatellite instability status were also determined in all tumors. RESULTS: The expressions of HGM and MUC6, representing gastric or gastric and intestinal mixed phenotypes, were significantly associated with high grade atypia in the 63 gastric adenomas. Among the 133 early differentiated-type carcinomas, HGM expression was significantly associated with mixed-type (with an undifferentiated-type component) tumors and lymph node metastasis. MUC2 expression was inversely associated with submucosal invasion. A multivariate analysis revealed that gastric adenomas were significantly associated with the intestinal phenotype and were inversely associated with p53 mutation compared with early differentiated-type carcinomas. Among all 196 tumors, APC mutation was significantly associated with CD10 expression and the intestinal phenotype and was inversely associated with the expressions of HGM and MUC6. The microsatellite instability status was significantly associated with MUC6 expression. Malignant transformation from gastric adenoma to carcinoma was shown in 5 of the 24 follow up cases of gastric adenoma. The malignant transformation was significantly associated with the gastric and intestinal mixed phenotype and was inversely associated with APC mutation. No malignant transformation was found in intestinal phenotype gastric adenomas with APC mutation. CONCLUSIONS: Our present findings show that phenotypic marker expression is associated with tumor aggressiveness during the early stage of gastric differentiated-type tumors. Differences in the biological behavior of tumors with different phenotypes may result from differences in the genetic backgrounds during the incipient phase of gastric tumorigenesis. PMID- 17085662 TI - Effect of chemotherapy on skeletal health in male survivors from testicular cancer and lymphoma. AB - PURPOSE: There are concerns over the late effects of cancer therapy, including accelerated bone loss leading to increased risk of osteoporosis. Treatment related bone loss is well recognized in breast and prostate cancer, due to overt hypogonadism, but there has been little evaluation of the skeletal effects of chemotherapy alone in adults. This study assesses the extent of bone loss due to previous chemotherapy in men. EXPERIMENTAL DESIGN: The bone mineral density (BMD) of men who had received previously chemotherapy with curative intent for lymphoma or testicular cancers was compared with that of an age-matched population of men from a cancer control population that had not received chemotherapy. BMD was measured by dual-energy X-ray scanning. Additionally, measurement of sex hormones and the bone turnover markers N-telopeptide fragment of type I collagen and bone specific alkaline phosphatase were done. All statistical tests were two sided. RESULTS: One hundred fifteen chemotherapy-treated patients and 102 cancer controls were recruited. There was no statistical difference in BMD between the chemotherapy and control groups at either spine or hip and the mean BMD values in both groups were no lower than that of a reference population. There were no significant differences in estradiol, luteinizing hormone, and testosterone, but follicle-stimulating hormone values were significantly higher in the chemotherapy group (P=0.011). The mean values of NH2-terminal telopeptide fragment of type I collagen and bone-specific alkaline phosphatase were within the reference ranges. CONCLUSIONS: The absence of accelerated bone loss following chemotherapy is reassuring and suggests that standard dose cytotoxic chemotherapy has no lasting clinically important direct effects on bone metabolism. PMID- 17085663 TI - Long-term events in adult patients with clinical stage IA-IIA nonbulky Hodgkin's lymphoma treated with four cycles of doxorubicin, bleomycin, vinblastine, and dacarbazine and adjuvant radiotherapy: a single-institution 15-year follow-up. AB - PURPOSE: To report on long-term events after short doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) chemotherapy and adjuvant radiotherapy in favorable early-stage Hodgkin's lymphoma. EXPERIMENTAL DESIGN: We monitored late events and causes of death over 15 years (median follow-up, 120 months) in 120 patients with nonbulky stage IA-IIA Hodgkin's lymphoma, treated with four cycles of ABVD and limited radiotherapy. Pulmonary and cardiac function tests were done throughout the follow-up. Outcome measures included cause-specific mortality, standardized mortality ratio, and standardized incidence ratio for secondary neoplasia. RESULTS: Projected 15-year event-free and overall survival were 78% and 86%, and tumor mortality was 3%. Standardized mortality ratio was significantly higher than 1 for both males (2.8; P=0.029) and females (9.4; P=0.003). The risk of cardiovascular events at 5 and 12 years was 5.5% and 14%, with a median latent time of 67 months (range: 23-179 months) from the end of radiotherapy. Pulmonary toxicity developed in 8% of patients; all had received mediastinal irradiation and the median time from radiotherapy to pulmonary sequelae was 76 weeks (range: 50-123 weeks). The risk of secondary neoplasia at 5 and 12 years was 4% and 8%, respectively, with no cases of leukemia. Fertility was preserved. CONCLUSIONS: Long-term events were mostly related to radiotherapy; the role of short ABVD chemotherapy was very limited, as documented by fertility preservation and lack of secondary myelodysplasia/leukemia. A proportion of patients died from causes unrelated to disease progression and the excess mortality risk was mostly due to the occurrence of secondary neoplasms and cardiovascular diseases. A moderate dose reduction of radiotherapy from 40-44 Gy to 30-36 Gy did not decrease the risk of late complications; abolishing radiotherapy in nonbulky early-stage Hodgkin's lymphoma is being evaluated. PMID- 17085664 TI - Novel D761Y and common secondary T790M mutations in epidermal growth factor receptor-mutant lung adenocarcinomas with acquired resistance to kinase inhibitors. AB - PURPOSE: In patients whose lung adenocarcinomas harbor epidermal growth factor receptor (EGFR) tyrosine kinase domain mutations, acquired resistance to the tyrosine kinase inhibitors (TKI) gefitinib (Iressa) and erlotinib (Tarceva) has been associated with a second-site EGFR mutation, which leads to substitution of methionine for threonine at position 790 (T790M). We aimed to elucidate the frequency and nature of secondary EGFR mutations in patients with acquired resistance to TKI monotherapy. EXPERIMENTAL DESIGN: Tumor cells from patients with acquired resistance were examined for secondary EGFR kinase domain mutations by molecular analyses. RESULTS: Eight of 16 patients (50% observed rate; 95% confidence interval, 25-75%) had tumor cells with second-site EGFR mutations. Seven mutations were T790M and one was a novel D761Y mutation found in a brain metastasis. When combined with a drug-sensitive L858R mutation, the D761Y mutation modestly reduced the sensitivity of mutant EGFR to TKIs in both surrogate kinase and cell viability assays. In an autopsy case, the T790M mutation was found in multiple visceral metastases but not in a brain lesion. CONCLUSIONS: The T790M mutation is common in patients with acquired resistance. The limited spectrum of TKI-resistant mutations in EGFR, which binds to erlotinib in the active conformation, contrasts with a wider range of second-site mutations seen with acquired resistance to imatinib, which binds to ABL and KIT, respectively, in closed conformations. Collectively, our data suggest that the type and nature of kinase inhibitor resistance mutations may be influenced by both anatomic site and mode of binding to the kinase target. PMID- 17085665 TI - Flat dosing of carboplatin is justified in adult patients with normal renal function. AB - PURPOSE: The Calvert formula is a widely applied algorithm for the a priori dosing of carboplatin based on patients glomerular filtration rate (GFR) as accurately measured using the 51Cr-EDTA clearance. Substitution of the GFR in this formula by an estimate of creatinine clearance or GFR as calculated by formulae using serum creatinine (SCR; Cockcroft-Gault, Jelliffe, and Wright) is, however, routine clinical practice in many hospitals. The goal of this study was to validate this practice retrospectively in a large heterogeneous adult patient population. EXPERIMENTAL DESIGN: Concentration-time data of ultrafilterable platinum of 178 patients (280 courses, 3,119 samples) with different types of cancer receiving carboplatin-based chemotherapy in conventional and high doses were available. Data were described with a linear two-compartment population pharmacokinetic model. Relations between SCR-based formulae for estimating renal function and carboplatin clearance were investigated. RESULTS: None of the tested SCR-based estimates of renal function were relevantly related to the pharmacokinetic variables of carboplatin. Neither SCR (median, 51; range, 18-124 micromol/L) nor the estimated GFR using the three different formulae was related to carboplatin clearance. CONCLUSIONS: Our data do not support the application of modifications of the Calvert formula by estimating GFR from SCR in the a priori dosing of carboplatin in patients with relatively normal renal function (creatinine clearance, >50 mL/min). For targeted carboplatin exposures, the original Calvert formula, measuring GFR using the 51Cr-EDTA clearance, remains the method of choice. Alternatively, in patients with normal renal function, a flat dose based on the mean population carboplatin clearance should be administered. PMID- 17085666 TI - A phase I study of single administration of antibody-directed enzyme prodrug therapy with the recombinant anti-carcinoembryonic antigen antibody-enzyme fusion protein MFECP1 and a bis-iodo phenol mustard prodrug. AB - PURPOSE: Antibody-directed enzyme prodrug therapy is a two-stage treatment whereby a tumor-targeted antibody-enzyme complex localizes in tumor for selective conversion of prodrug. The purpose of this study was to establish optimal variables for single administration of MFECP1, a recombinant antibody-enzyme fusion protein of an anti-carcinoembryonic antigen single-chain Fv antibody and the bacterial enzyme carboxypeptidase G2 followed by a bis-iodo phenol mustard prodrug. MFECP1 is manufactured in mannosylated form to facilitate normal tissue elimination. EXPERIMENTAL DESIGN: Pharmacokinetic, biodistribution, and tumor localization studies were used to test the hypothesis that MFECP1 localizes in tumor and clears from normal tissue via the liver. Firstly, safety of MFECP1 and a blood concentration of MFECP1 that would avoid systemic prodrug activation were tested. Secondly, dose escalation of prodrug was done. Thirdly, the dose of MFECP1 and timing of prodrug administration were optimized. RESULTS: MFECP1 was safe and well tolerated, cleared rapidly via the liver, and was less immunogenic than previously used products. Eighty-fold dose escalation from the starting dose of prodrug was carried out before dose-limiting toxicity occurred. Confirmation of the presence of enzyme in tumor and DNA interstrand cross-links indicating prodrug activation were obtained for the optimal dose and time point. A total of 28 of 31 patients was evaluable for response, the best response being a 10% reduction of tumor diameter, and 11 of 28 patients had stable disease. CONCLUSIONS: Optimal conditions for effective therapy were established. A study testing repeat treatment is currently being undertaken. PMID- 17085667 TI - Increased safety with preserved antitumoral efficacy on hepatocellular carcinoma with dual-regulated oncolytic adenovirus. AB - PURPOSE: A dual-regulated adenovirus variant CNHK500, in which human telomerase reverse transcriptase promoter drove the adenovirus 5 (Ad5) E1a gene and hypoxia response promoter controlled the E1b gene, was engineered. This virus has broad anticancer spectrum and higher specificity compared with mono-regulated adenovirus CNHK300. The objective of the current study is to show its antitumor selectivity and therapeutic potential. EXPERIMENTAL DESIGN: The antitumor specificity of human telomerase reverse transcriptase and hypoxia response promoters was evaluated in a panel of tumor and normal cells. Under the control of these promoters, the tumor-selective expression of E1a and E1b genes was evaluated. Further in vitro antitumor specificity and potency of this virus were characterized by viral replication and 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay. Subsequently, hepatocellular carcinoma xenografts were established to evaluate CNHK500 antitumor efficacy in vivo by different routes of virus administration and different dosages. RESULTS: Human telomerase reverse transcriptase and hypoxia response promoters were activated in a tumor-selective manner or under hypoxia treatment in a broad panel of cells. Selective adenoviral early gene expression, efficient viral replication, and oncolysis were observed in all tested cancer cells with more attenuated replication capacity in normal cells. Significant regression of hepatocellular carcinoma xenografts and prolonged survival were observed by either i.t. or i.v. administration. CONCLUSIONS: CNHK500 greatly reduced side effects in normal cells via dual control of adenoviral essential genes while still preserving potent antitumor efficacy on broad-spectrum cancer cells in vitro and in vivo. It can be used as a powerful therapeutic agent not only for liver cancers but also for other solid tumors. PMID- 17085668 TI - 2-methoxyestradiol, an endogenous mammalian metabolite, radiosensitizes colon carcinoma cells through c-Jun NH2-terminal kinase activation. AB - PURPOSE: 2-Methoxyestradiol (2ME), an estrogen metabolite, induces apoptosis in various cell types. We investigated whether 2ME pretreatment can radiosensitize colon adenocarcinoma cells. EXPERIMENTAL DESIGN: Radiosensitizing effects of 2ME were evaluated by cell death, clonogenic assay, nuclear fragmentation, and tumor progression of xenografts. Ionizing radiation-induced DNA damage was evaluated by histone H2AX phosphorylation and its foci. The c-Jun NH2-terminal kinase (JNK) activation was evaluated by anti-phosphorylated JNK antibody and inhibited by the JNK-specific inhibitor SP600125 or dominant-negative SEK1 expression. RESULTS: Clonogenic assays revealed that 2ME, but not estradiol, radiosensitized three colon carcinoma cells, DLD-1, HCT-8, and HCT-15, and strongly suppressed tumor progression of DLD-1 xenografts. Gene transfer-mediated Bcl-xL overexpression largely abolished both augmented apoptosis and reduced survival fractions. Pretreatment with 2ME enhanced H2AX phosphorylation, its foci, and phosphorylation of ATM kinase and delayed re-entry of cell cycle progression after ionizing radiation. Augmentation of both radiosensitivity and H2AX phosphorylation was substantially reduced by SP600125 or overexpression of a dominant-negative mutant SEK1. CONCLUSION: 2ME radiosensitized colon carcinoma cells through enhanced DNA damage via JNK activation, thereby representing a novel radiosensitizing therapy against colon cancer. PMID- 17085669 TI - Antileukemic activity of lysophosphatidic acid acyltransferase-beta inhibitor CT32228 in chronic myelogenous leukemia sensitive and resistant to imatinib. AB - PURPOSE: Lysophosphatidic acid acyltransferase (LPAAT)-beta catalyzes the conversion of lysophosphatidic acid to phosphatidic acid, an essential component of several signaling pathways, including the Ras/mitogen-activated protein kinase pathway. Inhibition of LPAAT-beta induces growth arrest and apoptosis in cancer cell lines, implicating LPAAT-beta as a potential drug target in neoplasia. EXPERIMENTAL DESIGN: In this study, we investigated the effects of CT32228, a specific LPAAT-beta inhibitor, on BCR-ABL-transformed cell lines and primary cells from patients with chronic myelogenous leukemia. RESULTS: CT32228 had antiproliferative activity against BCR-ABL-positive cell lines in the nanomolar dose range, evidenced by cell cycle arrest in G2-M and induction of apoptosis. Treatment of K562 cells with CT32228 led to inhibition of extracellular signal regulated kinase 1/2 phosphorylation, consistent with inhibition of mitogen activated protein kinase signaling. Importantly, CT32228 was highly active in cell lines resistant to the Bcr-Abl kinase inhibitor imatinib. Combination of CT32228 with imatinib produced additive inhibition of proliferation in cell lines with residual sensitivity toward imatinib. In short-term cultures in the absence of growth factors, CT32228 preferentially inhibited the growth of granulocyte macrophage colony-forming units from chronic myelogenous leukemia patients compared with healthy controls. CONCLUSION: These data establish LPAAT-beta as a potential drug target for the treatment of BCR-ABL-positive leukemias. PMID- 17085670 TI - Schedule-dependent synergy between the heat shock protein 90 inhibitor 17 (dimethylaminoethylamino)-17-demethoxygeldanamycin and doxorubicin restores apoptosis to p53-mutant lymphoma cell lines. AB - PURPOSE: Loss of p53 function impairs apoptosis induced by DNA-damaging agents used for cancer therapy. Here, we examined the effect of the heat shock protein 90 (HSP90) inhibitor 17-(dimethylaminoethylamino)-17-demethoxygeldanamycin (DMAG) on doxorubicin-induced apoptosis in lymphoma. We aimed to establish the optimal schedule for administration of both drugs in combination and the molecular basis for their interaction. EXPERIMENTAL DESIGN: Isogenic lymphoblastoid and nonisogenic lymphoma cell lines differing in p53 status were exposed to each drug or combination. Drug effects were examined using Annexin V, active caspase-3, cell cycle, and cytotoxicity assays. Synergy was evaluated by median effect/combination index. Protein expression and kinase inhibition provided insight into the molecular mechanisms of drug interaction. RESULTS: Presence of mutant p53 conferred increased survival to single agents. Nevertheless, DMAG showed synergistic toxicity with doxorubicin independently of p53 status. Synergy required exposure to doxorubicin before DMAG. DMAG-mediated down-regulation of CHK1, a known HSP90 client, forced doxorubicin-treated cells into premature mitosis followed by apoptosis. A CHK1 inhibitor, SB-218078, reproduced the effect of DMAG. Administration of DMAG before doxorubicin resulted in G1-S arrest and protection from apoptosis, leading to additive or antagonistic interactions that were exacerbated by p53 mutation. CONCLUSIONS: Administration of DMAG to doxorubicin-primed cells induced premature mitosis and had a synergistic effect on apoptosis regardless of p53 status. These observations provide a rationale for prospective clinical trials and stress the need to consider schedule of exposure as a critical determinant of the overall response when DMAG is combined with chemotherapeutic agents for the treatment of patients with relapsed/refractory disease. PMID- 17085671 TI - Synthetic microRNA designed to target glioma-associated antigen 1 transcription factor inhibits division and induces late apoptosis in pancreatic tumor cells. AB - PURPOSE: To determine whether the synthetic microRNAs (miRNA) could effectively target tumor cells we designed several miRNA complementary to glioma-associated antigen-1 (Gli-1) mRNA and investigated their ability to inhibit tumor cell proliferation. The sonic hedgehog pathway is an early and late mediator of tumorigenesis in epithelial cancers. Activation of sonic hedgehog signaling seems to precede transformation of tissue stem cells to cancerous stem cells, with the Gli-1 transcription factor functioning as a mediator of environmental signals. Inhibiting cancer cell proliferation by targeting the Gli-1 effector pathway is difficult to achieve by chemotherapeutic agents or short interfering RNA. EXPERIMENTAL DESIGN: We hypothesized that targeting the 3'-untranslated region of Gli-1 mRNA would effectively inhibit tumor cell proliferation. To test this hypothesis, we used synthetic miRNAs of our own design and corresponding duplex/small temporal RNAs by introducing three-nucleotide loops in the 3' untranslated region Gli-1 sequence of high GU content. RESULTS: We found that miRNA (Gli-1-miRNA-3548) and its corresponding duplex (Duplex-3548) significantly inhibited proliferation of Gli-1+ ovarian (SK-OV-3) and pancreatic (MiaPaCa-2) tumor cells. The miRNAs mediated delayed cell division and activation of late apoptosis in MiaPaCa-2 cells. This is the first demonstration of inhibition of pancreatic tumor cell division by designed miRNA. CONCLUSIONS: Gli-1 miRNAs should significantly add to the general understanding of the mechanisms of metastasis and contribute toward the design of better treatments for epithelial cancers. PMID- 17085672 TI - Hedgehog: an attribute to tumor regrowth after chemoradiotherapy and a target to improve radiation response. AB - PURPOSE: Despite aggressive chemotherapy, radiotherapy, surgery, or combination approaches, the survival rate of patients with esophageal cancer remains poor. Recent studies have suggested that constitutive activation of the Hedgehog (Hh) pathway in cancers of the digestive tract may contribute to the growth and maintenance of cancer. However, the relationship between Hh signaling and therapeutic response is unknown. EXPERIMENTAL DESIGN: The expression and temporal kinetics of Hh signaling and proliferation biomarkers after chemoradiotherapy were examined in esophageal tumor xenografts. Additionally, immunohistochemical analysis of Sonic Hh (Shh) and Gli-1 expression were done on residual tumors from patients who received neoadjuvant chemoradiotherapy followed by surgery. The ability of Shh signaling to induce proliferation in esophageal cell lines was determined. Expression of cell cycle checkpoint proteins was analyzed in cells in which Hh signaling was activated or inhibited. We further determined the effect of inhibiting Hh signaling in sensitizing esophageal tumors to radiation. RESULTS: We showed that the Shh signaling pathway was extensively activated in esophageal cancer xenografts and residual tumors after chemoradiotherapy and the temporal kinetics of Hh signaling preceded increases in proliferation biomarker expression and tumor size during tumor regrowth. We further showed that Hh pathway activity influences proliferation rates of esophageal cancer cell lines through up-regulation of the G1-cyclin-Rb axis. Additionally, we found that blocking Hh signaling enhanced radiation cytotoxicity of esophageal cancer cells. CONCLUSIONS: These results suggest that activation of the Hh pathway may promote tumor repopulation after chemoradiotherapy and contribute to chemoradiation resistance in esophageal cancers. PMID- 17085673 TI - Anti-vascular endothelial growth factor receptor-1 antagonist antibody as a therapeutic agent for cancer. AB - PURPOSE: Vascular endothelial growth factor receptor-1 (VEGFR-1) plays important roles in promotion of tumor growth by mediating cellular functions in tumor vascular endothelium and cancer cells. Blockade of VEGFR-1 activation has been shown to inhibit pathologic angiogenesis and tumor growth, implicating VEGFR-1 as a potential therapeutic target for the treatment of cancer. We have thus developed a VEGFR-1 antagonist human monoclonal antibody designated as IMC-18F1 and evaluated its antitumor activity in preclinical experimental models to show the therapeutic potential of the antibody for cancer treatment in clinic. EXPERIMENTAL DESIGN: Human IgG transgenic mice were used for generation of anti VEGFR-1 antibodies. Anti-VEGFR-1-specific blocking antibodies were identified using solid-phase binding and blocking assays. Inhibitory antitumor cell activity of IMC-18F1 was assessed in cell-based kinase and growth assays. Pharmacokinetic/pharmacodynamic studies were done to determine the association of antibody blood level with antitumor efficacy of the antibody in vivo. Antitumor efficacy of the anti-VEGFR-1 antibodies as monotherapy and in combination with cytotoxic agents was evaluated in human breast cancer xenograft models. RESULTS: A fully human neutralizing antibody, IMC-18F1, was shown to be a high-affinity (KD=54 pmol) inhibitor of VEGFR-1 ligand binding (VEGF-A, VEGF-B, and placental growth factor). IMC-18F1 inhibited ligand-induced intracellular activation of VEGFR-1 and mitogen-activated protein kinase signaling and prevented ligand stimulated in vitro growth of breast cancer cells. In vivo, IMC-18F1 suppressed the growth of human breast tumor xenografts in association with reduced mitogen activated protein kinase and Akt activation, reduced tumor cell proliferation, and increased tumor cell apoptosis. Pharmacokinetic/pharmacodynamic studies established a plasma elimination half-life of 5 days for IMC-18F1 and a steady state trough plasma therapeutic threshold of 88 microg/mL. Importantly, inhibition of mouse and human VEGFR-1 with MF1 and IMC-18F1, respectively, enhanced the antitumor efficacy of cytotoxic agents commonly used to treat breast cancer. CONCLUSIONS: Based on preclinical validation studies, IMC-18F1 anti-VEGFR 1 has potential to provide clinical benefit to cancer patients. PMID- 17085674 TI - Genetic variants of UGT1A6 influence risk of colorectal adenoma recurrence. AB - PURPOSE: The UDP glucuronosyltransferase 1A6 (UGT1A6) and cytochrome P450 2C9 (CYP2C9) enzymes participate in the metabolism of nonsteroidal anti-inflammatory drugs, endogenous substances, and carcinogens. Functional polymorphisms of UGT1A6 (T181A and R184S) and CYP2C9 (R144C and I359L) have been reported to modify the protective effect of aspirin on colorectal adenoma risk. We aimed to further investigate the effect of these genetic variants on the development of colorectal neoplasia. EXPERIMENTAL DESIGN: We examined the relationship between UGT1A6 and CYP2C9 genotype and colorectal adenoma recurrence in 546 patients participating in a randomized placebo-controlled aspirin intervention trial. RESULTS: Although colorectal adenoma recurrence was not significantly influenced by CYP2C9 genotype, carriers of variant UGT1A6 alleles were at significantly reduced risk of colorectal neoplasia recurrence [relative risk (RR), 0.68; 95% confidence interval (95% CI), 0.52-0.89]. This risk reduction was also evident when the analysis was confined to advanced neoplasia recurrence (RR, 0.71; 95% CI, 0.47 1.09). When patients were stratified by genotype and aspirin intervention, those with variant UGT1A6 alleles were at reduced recurrence risk irrespective of whether they received aspirin or placebo (RR, 0.62; 95% CI, 0.42-0.92 and RR, 0.63; 95% CI, 0.44-0.91, respectively). CONCLUSIONS: These findings confirm that UGT1A6 variants influence colorectal carcinogenesis independent of aspirin intake and suggest that they may have clinical value in secondary prevention programs for patients diagnosed with colorectal adenoma. PMID- 17085675 TI - Increasing renal mass improves survival in anephric rats following metanephros transplantation. AB - Renal failure and end-stage renal disease are prevalent diseases associated with high levels of morbidity and mortality, the preferred treatment for which is kidney transplantation. However, the gulf between supply and demand for kidneys remains high and is growing every year. A potential alternative to the transplantation of mature adult kidneys is the transplantation of the developing renal primordium, the metanephros. It has been shown previously, in rodent models, that transplantation of a metanephros can provide renal function capable of prolonging survival in anephric animals. The aim of the present study was to determine whether increasing the mass of transplanted tissue can prolong survival further. Embryonic day 15 rat metanephroi were transplanted into the peritoneum of anaesthetized adult rat recipients. Twenty-one days later, the transplanted metanephroi were anastomosed to the recipient's urinary system, and 35 days following anastomosis the animal's native renal mass was removed. Survival times and composition of the excreted fluid were determined. Rats with single metanephros transplants survived 29 h longer than anephric controls (P < 0.001); animals with two metanephroi survived 44 h longer (P < 0.001). A dilute urine was formed, with low concentrations of sodium, potassium and urea; potassium and urea concentrations were elevated in terminal serum samples, but sodium concentration and osmolality were comparable to control values. These data show that survival time is proportional to the mass of functional renal tissue. While transplanted metanephroi cannot currently provide life-sustaining renal function, this approach may have therapeutic benefit in the future. PMID- 17085676 TI - Increased sympathetic activity in rats submitted to chronic intermittent hypoxia. AB - Long-term exposure to intermittent hypoxia may lead to important cardiovascular dysfunctions, such as hypertension. Rodent models of chronic intermittent hypoxia (CIH) have been used to study the mechanisms underlying the increase in mean arterial pressure (MAP) observed after exposure to CIH. Several studies suggest that the hypertension of rats submitted to CIH is associated with an increase in sympathetic activity. However, there are no studies documenting the direct measurement of sympathetic activity in conscious freely moving rats exposed to CIH. Therefore, the present study aimed to evaluate whether or not the increase of MAP in rats exposed to CIH is associated with an increase in sympathetic activity. To reach this goal, we analysed the effect of ganglionic blockade on baseline MAP as well as the plasma levels of catecholamines. Rats submitted to CIH (fractional inspired O(2) of 6%, for 40 s in every 9 min, 8 h day(-1)) for 35 days (n = 31) exhibited a significant increase in MAP compared with control rats (n = 28) maintained under normoxia (112 +/- 2 versus 103 +/- 1 mmHg, P = 0.0003). The injection of the ganglionic blocker hexamethonium resulted in a similar fall in MAP in CIH and control groups (-46 +/- 2 versus -41 +/- 3 mmHg). However, hexamethonium after previous antagonism of the angiotensin II type 1 (AT(1)) receptors with losartan produced a larger decrease in MAP in the CIH than in the control group (-58 +/- 2 versus -50 +/- 2 mmHg, P = 0.0165). The injection of losartan itself produced no major changes in the baseline MAP in both groups. The measurement of plasma catecholamines showed an increase in plasma noradrenaline (10.12 +/- 0.90 versus 4.74 +/- 0.32 ng ml(-1), P = 0.0042) in rats exposed to CIH compared with control rats. These data provide strong evidence to support the concept that rats submitted to CIH exhibit an increase in sympathetic activity, which seems to be determinant in the maintenance of hypertension in this experimental model. PMID- 17085677 TI - Time course of the renal functional response to partial nephrectomy: measurements in conscious rats. AB - Previous investigations into the functional responses of the surviving nephrons following reductions in renal mass have been performed largely in anaesthetized animals and have taken little account of how the compensatory changes develop with time. The present study has assessed a method for determining glomerular filtration rate (GFR) in unrestrained, uncatheterized, conscious rats (plasma disappearance of (99m)Tc-diethylenetriamene pentaacetic acid (DTPA)) and has used this method to document the time course of the changes in GFR over a 32 day period following uninephrectomy or 5/6 nephrectomy. Concurrent measurements of excretion rates and of the clearance of lithium (the latter being an index of end proximal fluid delivery) provided information on changes in overall tubular function and segmental reabsorption. After uninephrectomy, the GFR of the remaining kidney (compared with that of a single kidney of sham-operated animals) increased maximally (by approximately 50%) within 8 days; after 5/6 nephrectomy, the increase in the GFR of the remnant kidney was maximal (at approximately 300%) within 16 days. Overall excretion rates of sodium and potassium were well maintained in partially nephrectomized animals throughout the period of study, while the excretion of water increased (by approximately 30% after uninephrectomy and by approximately 120% after 5/6 nephrectomy), partly as a result of the compensatory increases in GFR but mainly as a consequence of moderate (after uninephrectomy) or marked (after 5/6 nephrectomy) reductions in fractional reabsorption. During the early period after 5/6 nephrectomy, potassium excretion sometimes exceeded the filtered load, indicating net secretion. Lithium clearance data indicated that the changes in tubular function after 5/6 nephrectomy include a reduction in fractional reabsorption in the proximal tubule, whereas after uninephrectomy any such effect on the proximal tubule is minor and transient. PMID- 17085678 TI - Sleep and the metabolic syndrome. AB - The metabolic syndrome represents a clustering of several interrelated risk factors of metabolic origin that are thought to increase cardiovascular risk. It is still uncertain whether this clustering results from multiple underlying risk factors or whether it has a single cause. One metabolic abnormality that may underlie several clinical characteristics of the metabolic syndrome is insulin resistance. This review discusses the evidence that sleep disturbances (obstructive sleep apnoea, sleep deprivation and shift work) may independently lead to the development of both insulin resistance and individual clinical components of the metabolic syndrome. The converse may also be true, in that metabolic abnormalities associated with the metabolic syndrome and insulin resistance may potentially exacerbate sleep disorders. The notion that sleep disturbances exert detrimental metabolic effects may help explain the increasing prevalence of the metabolic syndrome and insulin resistance in the general population and may have important implications for population-based approaches to combat the increasing epidemic of metabolic and cardiovascular disease. PMID- 17085679 TI - Intraspecific communication through chemical signals in female mice: reinforcing properties of involatile male sexual pheromones. AB - In rodents, social and reproductive behaviors critically depend on chemical signals, including sexual pheromones that have been suggested (but not demonstrated) to be rewarding. In this work, we analyze this issue by studying the chemoinvestigatory behavior of adult female mice (without experience with male-derived chemicals) toward 1) the synthetic odorant citralva, 2) bedding soiled by different conspecifics (females, males, and castrated males), and 3) volatiles derived from bedding soiled by males and castrated males (confronted in 2-choice tests). We also study whether these chemical signals are able to induce conditioned place preference, a reliable test for rewarding properties of stimuli. The results show that involatile, male-derived chemicals elicit an intense and sustained chemoinvestigation and, more importantly, are the only tested chemical signals that induce conditioned place preference. In contrast, volatile, male-derived chemicals are not significantly chemoinvestigated. Bedding soiled by castrated males induces a transient chemoinvestigation, likely directed to steroid-independent, biologically relevant chemical signals, whereas the intense chemoinvestigation of female-soiled bedding shows a slow habituation. Finally, females did not explore significantly citralva-odorized bedding. The present work constitutes the first demonstration of the unconditioned reinforcing properties of involatile (likely detected by the vomeronasal organ) steroid dependent chemical signals in mammals. PMID- 17085680 TI - Molecular and clinical characterisation of three Spanish families with maternally inherited non-syndromic hearing loss caused by the 1494C->T mutation in the mitochondrial 12S rRNA gene. AB - Mutations in the 12S rRNA gene of the mitochondrial genome are responsible for maternally inherited non-syndromic hearing loss (NSHL), and for increased susceptibility to the ototoxicity of aminoglycoside antibiotics. Among these mutations, 1555A-->G is the most prevalent in all populations tested so far. Recently, the 1494C-->T mutation was reported in two large Chinese pedigrees with maternally inherited NSHL. In this study, sequencing of the 12S rRNA gene in a Spanish family with maternally inherited NSHL showed the presence of the 1494C- >T mutation. An additional screening of 1339 unrelated Spanish patients with NSHL allowed the authors to find two other families with the mutation. Audiological data were obtained from 17 confirmed 1494C-->T carriers, which showed that the hearing loss was sensorineural, bilateral and symmetrical, with a remarkable variability in age of onset and severity. Three carriers were asymptomatic. Three affected carriers had a history of treatment with aminoglycoside antibiotics. The mitochondrial genome of one affected person from each of these three families was entirely sequenced, and it was established that they belong to different mitochondrial haplogroups (H, U5b, U6a). The study results further support the pathogenic role of 1494C-->T on hearing, and show that this mutation can be found in different Caucasian mitochondrial DNA backgrounds. PMID- 17085681 TI - Identification of 14 novel mutations in the long isoform of USH2A in Spanish patients with Usher syndrome type II. AB - Mutations in USH2A gene have been shown to be responsible for Usher syndrome type II, an autosomal recessive disorder characterised by hearing loss and retinitis pigmentosa. USH2A was firstly described as consisting of 21 exons, but 52 novel exons at the 3' end of the gene were recently identified. In this report, a mutation analysis of the new 52 exons of USH2A gene was carried out in 32 unrelated patients in which both disease-causing mutations could not be found after the screening of the first 21 exons of the USH2A gene. On analysing the new 52 exons, fourteen novel mutations were identified in 14 out of the 32 cases studied, including 7 missense, 5 frameshift, 1 duplication and a putative splice site mutation. PMID- 17085682 TI - A large germline deletion in the Chek2 kinase gene is associated with an increased risk of prostate cancer. AB - BACKGROUND: Germline mutations in the Chek2 kinase gene (CHEK2) have been associated with a range of cancer types. Recently, a large deletion of exons 9 and 10 of CHEK2 was identified in several unrelated patients with breast cancer of Czech or Slovak origin. The geographical and ethnic extent of this founder allele has not yet been determined. PARTICIPANTS AND METHODS: We assayed for the presence of this deletion, and of three other CHEK2 founder mutations, in 1864 patients with prostate cancer and 5496 controls from Poland. RESULTS: The deletion was detected in 24 of 5496 (0.4%) controls from the general population, and is the most common CHEK2 truncating founder allele in Polish patients. The deletion was identified in 15 of 1864 (0.8%) men with unselected prostate cancer (OR 1.9; 95% CI 0.97 to 3.5; p = 0.09) and in 4 of 249 men with familial prostate cancer (OR 3.7; 95% CI 1.3 to 10.8; p = 0.03). These ORs were similar to those associated with the other truncating mutations (IVS2+1G-->A, 1100delC). CONCLUSION: A large deletion of exons 9 and 10 of CHEK2 confers an increased risk of prostate cancer in Polish men. The del5395 founder deletion might be present in other Slavic populations, including Ukraine, Belarus, Russia, Baltic and Balkan countries. It will be of interest to see to what extent this deletion is responsible for the burden of prostate cancer in other populations. PMID- 17085683 TI - The exoribonuclease XRN4 is a component of the ethylene response pathway in Arabidopsis. AB - EXORIBONUCLEASE4 (XRN4), the Arabidopsis thaliana homolog of yeast XRN1, is involved in the degradation of several unstable mRNAs. Although a role for XRN4 in RNA silencing of certain transgenes has been reported, xrn4 mutant plants were found to lack any apparent visible phenotype. Here, we show that XRN4 is allelic to the unidentified components of the ethylene response pathway ETHYLENE INSENSITIVE5/ACC-INSENSITIVE1 (EIN5/AIN1) and EIN7. xrn4 mutant seedlings are ethylene-insensitive as a consequence of the upregulation of EIN3 BINDING F-BOX PROTEIN1 (EBF1) and EBF2 mRNA levels, which encode related F-box proteins involved in the turnover of EIN3 protein, a crucial transcriptional regulator of the ethylene response pathway. Epistasis analysis placed XRN4/EIN5/AIN1 downstream of CTR1 and upstream of EBF1/2. XRN4 does not appear to regulate ethylene signaling via an RNA-INDUCED SILENCING COMPLEX-based RNA silencing mechanism but acts by independent means. The identification of XRN4 as an integral new component in ethylene signaling adds RNA degradation as another posttranscriptional process that modulates the perception of this plant hormone. PMID- 17085684 TI - Experimental reconstruction of functional gene transfer from the tobacco plastid genome to the nucleus. AB - Eukaryotic cells arose through the uptake of free-living bacteria by endosymbiosis and their gradual conversion into organelles (plastids and mitochondria). Capture of the endosymbionts was followed by massive translocation of their genes to the genome of the host cell. How genes were transferred from the (prokaryotic) organellar genome to the (eukaryotic) nuclear genome and how the genes became functional in their new eukaryotic genetic environment is largely unknown. Here, we report the successful experimental reconstruction of functional gene transfer between an organelle and the nucleus, a process that normally occurs only on large evolutionary timescales. In consecutive genetic screens, we first transferred a chloroplast genome segment to the nucleus and then selected for gene activation in the nuclear genome. We show that DNA mediated gene transfer can give rise to functional nuclear genes if followed by suitable rearrangements in the nuclear genome. Acquisition of gene function involves (1) transcriptional activation by capture of the promoter of an upstream nuclear gene and (2) utilization of AT-rich noncoding sequences downstream of the plastid gene as RNA cleavage and polyadenylation sites. Our results reveal the molecular mechanisms of how organellar DNA transferred to the nucleus gives rise to functional genes and reproduce in the laboratory a key process in the evolution of eukaryotic cells. PMID- 17085685 TI - The crystal structure of Arabidopsis thaliana allene oxide cyclase: insights into the oxylipin cyclization reaction. AB - We describe the crystallization and structure elucidation of Arabidopsis thaliana allene oxide cyclase 2 (AOC2), a key enzyme in the biosynthesis of jasmonates. In a coupled reaction with allene oxide synthase, AOC2 releases the first cyclic and biologically active metabolite, 12-oxo-phytodienoic acid (OPDA). AOC2 (AT3G25770) folds into an eight-stranded antiparallel beta-barrel with a C-terminal partial helical extension. The protein forms a hydrophobic binding cavity with two distinct polar patches. AOC2 is trimeric in crystals, in vitro and in planta. Based on the observed folding pattern, we assigned AOC2 as a low molecular weight member of the lipocalin family with enzymatic activity in plants. We determined the binding position of the competitive inhibitor vernolic acid (a substrate analog) in the binding pocket. Based on models for bound substrate 12,13-epoxy 9,11,15-octadecatrienoic acid and product OPDA, we propose a reaction scheme that explains the influence of the C15 double bond on reactivity. Reaction is promoted by anchimeric assistance through a conserved Glu residue. The transition state with a pentadienyl carbocation and an oxyanion is stabilized by a strongly bound water molecule and favorable pi-pi interactions with aromatic residues in the cavity. Stereoselectivity results from steric restrictions to the necessary substrate isomerizations imposed by the protein. PMID- 17085686 TI - Arabidopsis GCN5, HD1, and TAF1/HAF2 interact to regulate histone acetylation required for light-responsive gene expression. AB - We previously showed that Arabidopsis thaliana histone acetyltransferase TAF1/HAF2 is required for the light regulation of growth and gene expression, and we show here that histone acetyltransferase GCN5 and histone deacetylase HD1/HDA19 are also involved in such regulation. Mutation of GCN5 resulted in a long-hypocotyl phenotype and reduced light-inducible gene expression, whereas mutation of HD1 induced opposite effects. The double mutant gcn5 hd1 restored a normal photomorphogenic phenotype. By contrast, the double mutant gcn5 taf1 resulted in further loss of light-regulated gene expression. gcn5 reduced acetylation of histones H3 and H4, mostly on the core promoter regions, whereas hd1 increased acetylation on both core and more upstream promoter regions. GCN5 and TAF1 were both required for H3K9, H3K27, and H4K12 acetylation on the target promoters, but H3K14 acetylation was dependent only on GCN5. Interestingly, gcn5 taf1 had a cumulative effect mainly on H3K9 acetylation. On the other hand, hd1 induced increased acetylation on H3K9, H3K27, H4K5, and H4K8. GCN5 was also shown to be directly associated with the light-responsive promoters. These results suggest that acetylation of specific histone Lys residues, regulated by GCN5, TAF1, and HD1, is required for light-regulated gene expression. PMID- 17085687 TI - Silencing threonine deaminase and JAR4 in Nicotiana attenuata impairs jasmonic acid-isoleucine-mediated defenses against Manduca sexta. AB - Threonine deaminase (TD) catalyzes the conversion of Thr to alpha-keto butyrate in Ile biosynthesis; however, its dramatic upregulation in leaves after herbivore attack suggests a role in defense. In Nicotiana attenuata, strongly silenced TD transgenic plants were stunted, whereas mildly silenced TD transgenic plants had normal growth but were highly susceptible to Manduca sexta attack. The herbivore susceptibility was associated with the reduced levels of jasmonic acid-isoleucine (JA-Ile), trypsin proteinase inhibitors, and nicotine. Adding [(13)C(4)]Thr to wounds treated with oral secretions revealed that TD supplies Ile for JA-Ile synthesis. Applying Ile or JA-Ile to the wounds of TD-silenced plants restored herbivore resistance. Silencing JASMONATE-RESISTANT4 (JAR4), the N. attenuata homolog of the JA-Ile-conjugating enzyme JAR1, by virus-induced gene silencing confirmed that JA-Ile plays important roles in activating plant defenses. TD may also function in the insect gut as an antinutritive defense protein, decreasing the availability of Thr, because continuous supplementation of TD-silenced plants with large amounts (2 mmol) of Thr, but not Ile, increased M. sexta growth. However, the fact that the herbivore resistance of both TD- and JAR-silenced plants was completely restored by signal quantities (0.6 mumol) of JA-Ile treatment suggests that TD's defensive role can be attributed more to signaling than to antinutritive defense. PMID- 17085688 TI - Removal of astrovirus from water and sewage treatment plants, evaluated by a competitive reverse transcription-PCR. AB - Quantification of human astrovirus genogroups A and B was undertaken with sewage and water samples, collected from the Greater Cairo area in Egypt from November 1998 to October 1999, by a competitive reverse transcription (RT)-PCR with an internal control. The number of RNA copies of genogroup A/liter in quantifiable samples ranged from 3.4 x 10(3) to 5.6 x 10(6) in raw sewage and from 3.4 x 10(3) to 1.1 x 10(4) in treated effluents, while the number of infectious units per liter in these samples as determined by cell culture RT-PCR (CC-RT-PCR U/liter) ranged from 3.3 x 10(1) to 3.3 x 10(3) in raw sewage and was 3.3 x 10(0) in treated effluents. On the other hand, the number of RNA copies/liter in quantifiable genogroup B samples ranged from 1.1 x 10(4) to 8.7 x 10(6) in raw sewage and from 1.1 x 10(3) to 6.2 x 10(5) in treated effluents, while the number of infectious units ranged from 3.3 x 10(1) to 3.3 x 10(5) CC-RT-PCR U/liter in raw sewage and from 3.3 x 10(1) to 3.3 x 10(2) CC-RT-PCR U/liter in treated effluents. These higher numbers of both RNA copies/liter and infectious particles of genogroup B may indicate the emergence of genogroup B in the area. Additionally, genogroup B astrovirus exhibited a higher resistance to removal treatments with regard to the number of RNA copies per ml. When the equipment for real-time approaches is unavailable, a competitive PCR or RT-PCR with an internal control may be employed for virus quantification in validations of the efficiency of virus removal treatments. PMID- 17085689 TI - Longitudinal study of Finnish Campylobacter jejuni and C. coli isolates from humans, using multilocus sequence typing, including comparison with epidemiological data and isolates from poultry and cattle. AB - We describe a study on the application of multilocus sequence typing for the analysis of Campylobacter jejuni and C. coli isolates from human domestically acquired infections in the Helsinki-Uusimaa area of Finland in 1996, 2002, and 2003. In addition, isolates from poultry meat and fecal samples of cattle from the seasonal peak (July to September) in 2003 were included in the study. In total, 361 Finnish C. jejuni and C. coli strains were typed. Sequence type 45 (ST 45) (45%), ST-21 (21%), and ST-677 (11%) clonal complexes were the most prevalent. The ST-45 and ST-677 complexes were overrepresented in comparison with previous studies. The longitudinal study revealed an association between C. coli (ST-828 complex) infection and elderly patients (>/=60 years). Analysis of exposure factors, determined by a previous case-control study conducted during the seasonal peak in 2002, revealed that the ST-48 complex was significantly (P < 0.05) associated with the tasting or eating of raw minced meat. New and unassigned STs were associated with swimming in natural bodies of water, whereas the ST-677 complex was related to drinking nonchlorinated water from a small water plant or water from natural sources. The ST-45 complex was associated with contact with pet cats and dogs. In 2003, ST-45 occurrence was significantly associated with poultry whereas ST-50 was associated with isolates from humans. In contrast, ST-53, ST-58, ST-61, and ST-883 were significantly associated with isolates from cattle. Further studies are needed to reveal the significance of the observed associations. PMID- 17085690 TI - Shiga toxin and Shiga toxin-encoding phage do not facilitate Escherichia coli O157:H7 colonization in sheep. AB - Isogenic strains of Escherichia coli O157:H7, missing either stx(2) or the entire Stx2-encoding phage, were compared with the parent strain for their abilities to colonize sheep. The absence of the phage or of the Shiga toxin did not significantly impact the magnitude or duration of shedding of E. coli O157:H7. PMID- 17085691 TI - Molecular analysis of the subgingival microbiota in health and disease. AB - This investigation provides molecular analyses of the periodontal microbiota in health and disease. Subgingival samples from 47 volunteers with healthy gingivae or clinically diagnosed chronic periodontitis were characterized by PCR denaturing gradient gel electrophoresis (DGGE) with primers specific for the V2 V3 region of the eubacterial 16S rRNA gene. A hierarchical dendrogram was constructed from band patterns. All unique PCR amplicons (DGGE bands) were sequenced for identity. Samples were also analyzed for the presence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythensis by multiplex PCR. Associations of patient age, gender, and smoking status together with the presence of each unique band and putative periodontal pathogens with disease were assessed by logistic regression. Periodontal pockets were colonized by complex eubacterial communities (10 to 40 distinct DGGE bands) with substantial individual variation in the community profile. Species diversity in health and disease was determined by the Shannon-Weaver index of diversity and compared by the Mann-Whitney U test. Sequence analyses of DGGE amplicons indicated the occurrence of many nontypical oral species and eubacteria previously associated with this environment. With the exception of T. forsythensis, the putative pathogens were not detected by DGGE. Multiplex PCR, however, detected T. forsythensis, A. actinomycetemcomitans, and P. gingivalis in 9% 16%, and 29% of the patients with disease, respectively. The presence of A. actinomycetemcomitans was significantly associated with disease (P < 0.01). Statistical analyses indicated that the presence of Treponema socranskii and Pseudomonas sp. was a significant predictor of disease (P < 0.05) and that there was no significant difference (P > 0.05) in terms of eubacterial species diversity between health and disease. PMID- 17085692 TI - Rhizobium etli USDA9032 engineered to produce a phenazine antibiotic inhibits the growth of fungal pathogens but is impaired in symbiotic performance. AB - Phenazine production was engineered in Rhizobium etli USDA9032 by the introduction of the phz locus of Pseudomonas chlororaphis O6. Phenazine-producing R. etli was able to inhibit the growth of Botrytis cinerea and Fusarium oxysporum in vitro. Black bean inoculated with phenazine-producing R. etli produced brownish Fix(-) nodules. PMID- 17085693 TI - Sources of variation in the ampicillin-resistant Escherichia coli concentration in the feces of organic broiler chickens. AB - Currently, there are limited published data for the population dynamics of antimicrobial-resistant commensal bacteria. This study was designed to evaluate both the proportions of the Escherichia coli populations that are resistant to ampicillin at the level of the individual chicken on commercial broiler farms and the feasibility of obtaining repeated measures of fecal E. coli concentrations. Short-term temporal variation in the concentration of fecal E. coli was investigated, and a preliminary assessment was made of potential factors involved in the shedding of high numbers of ampicillin-resistant E. coli by growing birds in the absence of the use of antimicrobial drugs. Multilevel linear regression modeling revealed that the largest component of random variation in log transformed fecal E. coli concentrations was seen between sampling occasions for individual birds. The incorporation of fixed effects into the model demonstrated that the older, heavier birds in the study were significantly more likely (P = 0.0003) to shed higher numbers of ampicillin-resistant E. coli. This association between increasing weight and high shedding was not seen for the total fecal E. coli population (P = 0.71). This implies that, in the absence of the administration of antimicrobial drugs, the proportion of fecal E. coli that was resistant to ampicillin increased as the birds grew. This study has shown that it is possible to collect quantitative microbiological data on broiler farms and that such data could make valuable contributions to risk assessments concerning the transfer of resistant bacteria between animal and human populations. PMID- 17085694 TI - Identification and quantification of methanogenic Archaea in adult chicken ceca. AB - By using molecular methods for the identification and quantification of methanogenic archaea in adult chicken ceca, 16S rRNA genes of 11 different phylotypes, 10 of which were 99% similar to Methanobrevibacter woesei, were found. Methanogen populations, as assessed by cultivation, and the 16S rRNA copy number were between 6.38 and 8.23 cells/g (wet weight) and 5.50 and 7.19 log(10)/g (wet weight), respectively. PMID- 17085695 TI - Lytic activity of recombinant bacteriophage phi11 and phi12 endolysins on whole cells and biofilms of Staphylococcus aureus. AB - The recombinant phi11 endolysin hydrolyzed heat-killed staphylococci as well as staphylococcal biofilms. Cell wall targeting appeared to be a prerequisite for lysis of whole cells, and the combined action of the endopeptidase and amidase domains was necessary for maximum activity. In contrast, the phi12 endolysin was inactive and caused aggregation of the cells. PMID- 17085696 TI - Occurrence of virulence and antimicrobial resistance genes in Escherichia coli isolates from different aquatic ecosystems within the St. Clair River and Detroit River areas. AB - Although the number of Escherichia coli bacteria in surface waters can differ greatly between locations, relatively little is known about the distribution of E. coli pathotypes in surface waters used as sources for drinking or recreation. DNA microarray technology is a suitable tool for this type of study due to its ability to detect high numbers of virulence and antimicrobial resistance genes simultaneously. Pathotype, phylogenetic group, and antimicrobial resistance gene profiles were determined for 308 E. coli isolates from surface water samples collected from diverse aquatic ecosystems at six different sites in the St. Clair River and Detroit River areas. A higher frequency (48%) of E. coli isolates possessing virulence and antimicrobial resistance genes was observed in an urban site located downstream of wastewater effluent outfalls than in the other examined sites (average of 24%). Most E. coli pathotypes were extraintestinal pathogenic E. coli (ExPEC) pathotypes and belonged to phylogenetic groups B2 and D. The ExPEC pathotypes were found to occur across all aquatic ecosystems investigated, including riverine, estuarine, and offshore lake locations. The results of this environmental study using DNA microarrays highlight the widespread distribution of E. coli pathotypes in aquatic ecosystems and the potential public health threat of E. coli pathotypes originating from municipal wastewater sources. PMID- 17085697 TI - Titration and conditional knockdown of the prfB gene in Escherichia coli: effects on growth and overproduction of the recombinant mammalian selenoprotein thioredoxin reductase. AB - Release factor 2 (RF2), encoded by the prfB gene in Escherichia coli, catalyzes translational termination at UGA and UAA codons. Termination at UGA competes with selenocysteine (Sec) incorporation at Sec-dedicated UGA codons, and RF2 thereby counteracts expression of selenoproteins. prfB is an essential gene in E. coli and can therefore not be removed in order to increase yield of recombinant selenoproteins. We therefore constructed an E. coli strain with the endogenous chromosomal promoter of prfB replaced with the titratable P(BAD) promoter. Knockdown of prfB expression gave a bacteriostatic effect, while two- to sevenfold overexpression of RF2 resulted in a slightly lowered growth rate in late exponential phase. In a turbidostatic fermentor system the simultaneous impact of prfB knockdown on growth and recombinant selenoprotein expression was subsequently studied, using production of mammalian thioredoxin reductase as model system. This showed that lowering the levels of RF2 correlated directly with increasing Sec incorporation specificity, while also affecting total selenoprotein yield concomitant with a lower growth rate. This study thus demonstrates that expression of prfB can be titrated through targeted exchange of the native promoter with a P(BAD)-promoter and that knockdown of RF2 can result in almost full efficiency of Sec incorporation at the cost of lower total selenoprotein yield. PMID- 17085698 TI - Poly(3-hydroxybutyrate) granules at the early stages of formation are localized close to the cytoplasmic membrane in Caryophanon latum. AB - Localization of newly synthesized poly(3hydroxybutyrate) (PHB) granules was determined by confocal laser scanning fluorescence microscopy of Nile red-stained cells and by transmission electron microscopy (TEM). PHB granules of Nile red stained living cells of Caryophanon latum at the early stages of PHB accumulation were frequently found at or close to the cytoplasmic membrane. TEM analysis of the same culture revealed electron-translucent globular structures resembling PHB granules that were nonrandomly distributed in the cell lumen but were frequently found at or close to the cytoplasmic membrane. Immunogold labeling using PHB specific antiserum confirmed that the electron-translucent structures represented PHB granules. Electron microscopy examination of PHB granules after cell lysis revealed that PHB granules were often associated with membrane vesicles. Nonrandom localization of PHB granules was also found in Beijerinckia indica. Cells of this species harbored one PHB granule at each cell pole. Our results show that newly synthesized PHB granules often are close to or even in physical contact with the cytoplasmic membrane. Possible explanations for this unexpected finding and a hypothetical model of PHB granule formation in C. latum are discussed. PMID- 17085699 TI - Prevalence of Streptococcus suis genotypes in wild boars of Northwestern Germany. AB - Invasive serotype 2 (cps2+) strains of Streptococcus suis cause meningitis in pigs and humans. Four case reports of S. suis meningitis in hunters suggest transmission of S. suis through the butchering of wild boars. Therefore, the objective of this study was to investigate the prevalence of potentially human pathogenic S. suis strains in wild boars. S. suis was isolated from 92% of all tested tonsils (n=200) from wild boars. A total of 244 S. suis isolates were genotyped using PCR assays for the detection of serotype-specific genes, the hemolysin gene sly, and the virulence-associated genes mrp and epf. The prevalence of the cps2+ genotype among strains from wild boars was comparable to that of control strains from domestic pig carriers. Ninety-five percent of the cps2+ wild boar strains were positive for mrp, sly, and epf*, the large variant of epf. Interestingly, epf* was significantly more frequently detected in cps2+ strains from wild boars than in those from domestic pigs; epf* is also typically found in European S. suis isolates from humans, including a meningitis isolate from a German hunter. These results suggest that at least 10% of wild boars in Northwestern Germany carry S. suis strains that are potentially virulent in humans. Additional amplified fragment length polymorphism analysis supported this hypothesis, since homogeneous clustering of the epf* mrp+ sly+ cps2+ strains from wild boars with invasive human and porcine strains was observed. PMID- 17085700 TI - Autoinducer-2-producing protein LuxS, a novel salt- and chloride-induced protein in the moderately halophilic bacterium Halobacillus halophilus. AB - The moderately halophilic bacterium Halobacillus halophilus carries a homologue of LuxS, a protein involved in the activated methyl cycle and the production of autoinducer-2, which mediates quorum sensing between certain species. luxS of H. halophilus is part of an operon that encodes an S-adenosylmethionine-dependent methyltransferase, a cysteine synthase, and a beta-cystathionine lyase. Expression of luxS was growth phase dependent, with maximal expression in the mid exponential growth phase. In addition, transcription of luxS was strictly salt dependent; maximal mRNA concentrations were observed with 2.0 M NaCl in the growth medium. Chloride ions stimulated luxS transcription by a factor of three. Western blot analyses demonstrated a growth phase- and salinity-dependent production of LuxS. Moreover, cellular LuxS levels were strictly chloride dependent. Maximal accumulation of LuxS was observed at 0.5 to 1.0 M Cl(-) and depended on the salinity. PMID- 17085701 TI - Bacterial population dynamics in dairy waste during aerobic and anaerobic treatment and subsequent storage. AB - The objective of this study was to model a typical dairy waste stream, monitor the chemical and bacterial population dynamics that occur during aerobic or anaerobic treatment and subsequent storage in a simulated lagoon, and compare them to those of waste held without treatment in a simulated lagoon. Both aerobic and anaerobic treatment methods followed by storage effectively reduced the levels of total solids (59 to 68%), biological oxygen demand (85 to 90%), and sulfate (56 to 65%), as well as aerobic (83 to 95%), anaerobic (80 to 90%), and coliform (>99%) bacteria. However, only aerobic treatment reduced the levels of ammonia, and anaerobic treatment was more effective at reducing total sulfur and sulfate. The bacterial population structure of waste before and after treatment was monitored using 16S rRNA gene sequence libraries. Both treatments had unique effects on the bacterial population structure of waste. Aerobic treatment resulted in the greatest change in the type of bacteria present, with the levels of eight out of nine phyla being significantly altered. The most notable differences were the >16-fold increase in the phylum Proteobacteria and the approximately 8-fold decrease in the phylum Firmicutes. Anaerobic treatment resulted in fewer alterations, but significant decreases in the phyla Actinobacteria and Bacteroidetes, and increases in the phyla Planctomycetes, Spirochetes, and TM7 were observed. PMID- 17085702 TI - Evidence that accumulation of mutants in a biofilm reflects natural selection rather than stress-induced adaptive mutation. AB - The accumulation of mutant genotypes within a biofilm evokes the controversy over whether the biofilm environment induces adaptive mutation or whether the accumulation can be explained by natural selection. A comparison of the virulence of two strains of the dental pathogen Streptococcus mutans showed that rats infected with one of the strains accumulated a high proportion (average, 22%) of organisms that had undergone a deletion between two contiguous and highly homologous genes. To determine if the accumulation of deletion mutants was due to selection or to an increased mutation rate, accumulations of deletion mutants within in vitro planktonic and biofilm cultures and within rats inoculated with various proportions of deletion organisms were quantified. We report here that natural selection was the primary force behind the accumulation of the deletion mutants. PMID- 17085703 TI - Synthesis of N-acyl homoserine lactone analogues reveals strong activators of SdiA, the Salmonella enterica serovar Typhimurium LuxR homologue. AB - N-Acyl homoserine lactones (AHLs) are molecules that are synthesized and detected by many gram-negative bacteria to monitor the population density, a phenomenon known as quorum sensing. Salmonella enterica serovar Typhimurium is an exceptional species since it does not synthesize its own AHLs, while it does encode a LuxR homologue, SdiA, which enables this bacterium to detect AHLs that are produced by other species. To obtain more information about the specificity of the ligand binding by SdiA, we synthesized and screened a limited library of AHL analogues. We identified two classes of analogues that are strong activators of SdiA: the N-(3-oxo-acyl)-homocysteine thiolactones (3O-AHTLs) and the N-(3-oxo acyl)-trans-2-aminocyclohexanols. To our knowledge, this is the first report of compounds (the 3O-AHTLs) that are able to activate a LuxR homologue at concentrations that are lower than the concentrations of the most active AHLs. SdiA responds with greatest sensitivity to AHTLs that have a keto modification at the third carbon atom and an acyl chain that is seven or eight carbon atoms long. The N-(3-oxo-acyl)-trans-2-aminocyclohexanols were found to be less sensitive to deactivation by lactonase and alkaline pH than the 3O-AHTLs and the AHLs are. We also examined the activity of our library with LuxR of Vibrio fischeri and identified three new inhibitors of LuxR. Finally, we performed preliminary binding experiments which suggested that SdiA binds its activators reversibly. These results increase our understanding of the specificity of the SdiA-ligand interaction, which could have uses in the development of anti-quorum-sensing based antimicrobials. PMID- 17085704 TI - Mutation of Candida tropicalis by irradiation with a He-Ne laser to increase its ability to degrade phenol. AB - Candida tropicalis isolated from acclimated activated sludge was used in this study. Cell suspensions with 5 x 10(7) cells ml(-1) were irradiated by using a He Ne laser. After mutagenesis, the irradiated cell suspension was diluted and plated on yeast extract-peptone-dextrose (YEPD) medium. Plates with approximately 20 individual colonies were selected, and all individual colonies were harvested for phenol biodegradation. The phenol biodegradation stabilities for 70 phenol biodegradation-positive mutants, mutant strains CTM 1 to 70, ranked according to their original phenol biodegradation potentials, were tested continuously during transfers. Finally, mutant strain CTM 2, which degraded 2,600 mg liter(-1) phenol within 70.5 h, was obtained on the basis of its capacity and hereditary stability for phenol biodegradation. The phenol hydroxylase gene sequences were cloned in wild and mutant strains. The results showed that four amino acids were mutated by irradiation with a laser. In order to compare the activity of phenol hydroxylase in wild and mutant strains, their genes were expressed in Escherichia coli BL21(DE3) and enzyme activities were spectrophotometrically determined. It was clear that the activity of phenol hydroxylase was promoted after irradiation with a He-Ne laser. In addition, the cell growth and intrinsic phenol biodegradation kinetics of mutant strain CTM 2 in batch cultures were also described by Haldane's kinetic equation with a wide range of initial phenol concentrations from 0 to 2,600 mg liter(-1). The specific growth and degradation rates further demonstrated that the CTM 2 mutant strain possessed a higher capacity to resist phenol toxicity than wild C. tropicalis did. PMID- 17085706 TI - Procedure for rapid concentration and detection of enteric viruses from berries and vegetables. AB - Several hepatitis A virus (HAV) and norovirus (NV) outbreaks due to consumption of berries and vegetables have been reported during recent years. To facilitate the detection of enteric viruses that may be present on different fresh and frozen products, we developed a rapid and sensitive detection method for HAV, NV, and rotavirus (RV). Initial experiments focused on optimizing the composition of the elution buffer, improving the viral concentration method, and evaluating the performance of various extraction kits. Viruses were extracted from the food surface by a direct elution method in a glycine-Tris (pH 9.5) buffer containing 1% beef extract and concentrated by ultrafiltration. Occasionally, PCR inhibitors were present in the processed berry samples, which gave relatively poor detection limits. However, this problem was overcome by adding a pectinase treatment in the protocol, which markedly improved the sensitivity of the method. After optimization, this concentration method was applied in combination with real-time reverse transcription-PCR (RT-PCR) using specific primers in various types of berries and vegetables. The average detection limits were 1 50% tissue culture infective dose (TCID(50)), 54 RT-PCR units, and 0.02 TCID(50) per 15 g of food for HAV, NV, and RV, respectively. Based on our results, it is concluded that this procedure is suitable to detect and quantify enteric viruses within 6 h and can be applied for surveillance of enteric viruses in fresh and frozen products. PMID- 17085705 TI - A single-nucleotide-polymorphism-based multilocus genotyping assay for subtyping lineage I isolates of Listeria monocytogenes. AB - Listeria monocytogenes is a facultative intracellular pathogen responsible for food-borne disease with high mortality rates in humans and is the leading microbiological cause of food recalls. Lineage I isolates of L. monocytogenes are a particular public health concern because they are responsible for most sporadic cases of listeriosis and the vast majority of epidemic outbreaks. Rapid, reproducible, and sensitive methods for differentiating pathogens below the species level are required for effective pathogen control programs, and the CDC PulseNet Task Force has called for the development and validation of DNA sequence based methods for subtyping food-borne pathogens. Therefore, we developed a multilocus genotyping (MLGT) assay for L. monocytogenes lineage I isolates based on nucleotide variation identified by sequencing 23,251 bp of DNA from 22 genes distributed across seven genomic regions in 65 L. monocytogenes isolates. This single-well assay of 60 allele-specific probes captured 100% of the haplotype information contained in approximately 1.5 Mb of comparative DNA sequence and was used to reproducibly type a total of 241 lineage I isolates. The MLGT assay provided high discriminatory power (Simpson's index value, 0.91), uniquely identified isolates from the eight listeriosis outbreaks examined, and differentiated serotypes 1/2b and 4b as well as epidemic clone I (ECI), ECIa, and ECII. In addition, the assay included probes for a previously characterized truncation mutation in inlA, providing for the identification of a specific virulence-attenuated subtype. These results demonstrate that MLGT represents a significant new tool for use in pathogen surveillance, outbreak detection, risk assessment, population analyses, and epidemiological investigations. DNA sequences were deposited in the GenBank database under accession numbers DQ 812146 to DQ 812517, DQ 843664 to DQ 844598, and AY 512391 to AY 512502. PMID- 17085707 TI - Conjugative transfer facilitates stable maintenance of IncP-1 plasmid pKJK5 in Escherichia coli cells colonizing the gastrointestinal tract of the germfree rat. AB - Quantitative determination of IncP-1 plasmid loss from Escherichia coli cells colonizing the gastrointestinal tracts of germfree rats was achieved by flow cytometry. Results show that the plasmid's ability to conjugate counteracts plasmid loss and is thus an important mechanism for the stable maintenance of IncP-1 plasmids within the gastrointestinal environment. PMID- 17085708 TI - Longitudinal changes in the bacterial community composition of the Danube River: a whole-river approach. AB - The Danube River is the second longest river in Europe, and its bacterial community composition has never been studied before over its entire length. In this study, bacterial community composition was determined by denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified portions of the bacterial 16S rRNA gene from a total of 98 stations on the Danube River (73 stations) and its major tributaries (25 stations), covering a distance of 2,581 km. Shifts in the bacterial community composition were related to changes in environmental conditions found by comparison with physicochemical parameters (e.g., temperature and concentration of nutrients) and the concentration of chlorophyll a (Chl a). In total, 43 distinct DGGE bands were detected. Sequencing of selected bands revealed that the phylotypes were associated with typical freshwater bacteria. Apparent bacterial richness in the Danube varied between 18 and 32 bands and correlated positively with the concentration of P-PO(4) (r = 0.56) and negatively with Chl a (r = -0.52). An artificial neural network-based model explained 90% of the variation of apparent bacterial richness using the concentrations of N-NO(2) and P-PO(4) and the distance to the Black Sea as input parameters. Between the cities of Budapest and Belgrade, apparent bacterial richness was significantly lower than that of other regions of the river, and Chl a showed a pronounced peak. Generally, the bacterial community composition developed gradually; however, an abrupt and clear shift was detected in the section of the phytoplankton bloom. Large impoundments did not have a discernible effect on the bacterial community of the water column. In conclusion, the riverine bacterial community was largely influenced by intrinsic factors. PMID- 17085709 TI - Highly prevalent Coxiella sp. bacterium in the tick vector Amblyomma americanum. AB - Laboratory-reared and field-collected Amblyomma americanum ticks were hosts of a Coxiella sp. and a Rickettsia sp. While the Coxiella sp. was detected in 50 of 50 field-collected ticks, the Rickettsia sp. was absent from 32% of ticks. The Coxiella sp. showed evidence of a reduced genome and may be an obligate endosymbiont. PMID- 17085710 TI - Respiration and growth of Shewanella decolorationis S12 with an Azo compound as the sole electron acceptor. AB - The ability of Shewanella decolorationis S12 to obtain energy for growth by coupling the oxidation of various electron donors to dissimilatory azoreduction was investigated. This microorganism can reduce a variety of azo dyes by use of formate, lactate, pyruvate, or H(2) as the electron donor. Furthermore, strain S12 grew to a maximal density of 3.0 x 10(7) cells per ml after compete reduction of 2.0 mM amaranth in a defined medium. This was accompanied by a stoichiometric consumption of 4.0 mM formate over time when amaranth and formate were supplied as the sole electron acceptor and donor, respectively, suggesting that microbial azoreduction is an electron transport process and that this electron transport can yield energy to support growth. Purified membranous, periplasmic, and cytoplasmic fractions from S12 were analyzed, but only the membranous fraction was capable of reducing azo dyes with formate, lactate, pyruvate, or H(2) as the electron donor. The presence of 5 microM Cu(2+) ions, 200 microM dicumarol, 100 microM stigmatellin, and 100 microM metyrapone inhibited anaerobic azoreduction activity by both whole cells and the purified membrane fraction, showing that dehydrogenases, cytochromes, and menaquinone are essential electron transfer components for azoreduction. These results provide evidence that the microbial anaerobic azoreduction is linked to the electron transport chain and suggest that the dissimilatory azoreduction is a form of microbial anaerobic respiration. These findings not only expand the number of potential electron acceptors known for microbial energy conservation but also elucidate the mechanisms of microbial anaerobic azoreduction. PMID- 17085711 TI - Communities of archaea and bacteria in a subsurface radioactive thermal spring in the Austrian Central Alps, and evidence of ammonia-oxidizing Crenarchaeota. AB - Scanning electron microscopy revealed great morphological diversity in biofilms from several largely unexplored subterranean thermal Alpine springs, which contain radium 226 and radon 222. A culture-independent molecular analysis of microbial communities on rocks and in the water of one spring, the "Franz-Josef Quelle" in Bad Gastein, Austria, was performed. Four hundred fifteen clones were analyzed. One hundred thirty-two sequences were affiliated with 14 bacterial operational taxonomic units (OTUs) and 283 with four archaeal OTUs. Rarefaction analysis indicated a high diversity of bacterial sequences, while archaeal sequences were less diverse. The majority of the cloned archaeal 16S rRNA gene sequences belonged to the soil-freshwater-subsurface (1.1b) crenarchaeotic group; other representatives belonged to the freshwater-wastewater-soil (1.3b) group, except one clone, which was related to a group of uncultivated Euryarchaeota. These findings support recent reports that Crenarchaeota are not restricted to high-temperature environments. Most of the bacterial sequences were related to the Proteobacteria (alpha, beta, gamma, and delta), Bacteroidetes, and Planctomycetes. One OTU was allied with Nitrospina sp. (delta-Proteobacteria) and three others grouped with Nitrospira. Statistical analyses suggested high diversity based on 16S rRNA gene analyses; the rarefaction plot of archaeal clones showed a plateau. Since Crenarchaeota have been implicated recently in the nitrogen cycle, the spring environment was probed for the presence of the ammonia monooxygenase subunit A (amoA) gene. Sequences were obtained which were related to crenarchaeotic amoA genes from marine and soil habitats. The data suggested that nitrification processes are occurring in the subterranean environment and that ammonia may possibly be an energy source for the resident communities. PMID- 17085712 TI - Potential of the Bacillus thuringiensis toxin reservoir for the control of Lobesia botrana (Lepidoptera: Tortricidae), a major pest of grape plants. AB - The potential of Bacillus thuringiensis Cry proteins to control the grape pest Lobesia botrana was explored by testing first-instar larvae with Cry proteins belonging to the Cry1, Cry2, and Cry9 groups selected for their documented activities against Lepidoptera. Cry9Ca, a toxin from B. thuringiensis, was the protein most toxic to L. botrana larvae, followed in decreasing order by Cry2Ab, Cry1Ab, Cry2Aa, and Cry1Ia7, with 50% lethal concentration values of 0.09, 0.1, 1.4, 3.2, and 8.5 microg/ml of diet, respectively. In contrast, Cry1Fa and Cry1JA were not active at the assayed concentration (100 microg/ml). In vitro binding and competition experiments showed that none of the toxins tested (Cry1Ia, Cry2Aa, Cry2Ab, and Cry9C) shared binding sites with Cry1Ab. We conclude that either Cry1Ia or Cry9C could be used in combination with Cry1Ab to control this pest, either as the active components of B. thuringiensis sprays or expressed together in transgenic plants. PMID- 17085713 TI - Increased insect virulence in Beauveria bassiana strains overexpressing an engineered chitinase. AB - Entomopathogenic fungi are currently being used for the control of several insect pests as alternatives or supplements to chemical insecticides. Improvements in virulence and speed of kill can be achieved by understanding the mechanisms of fungal pathogenesis and genetically modifying targeted genes, thus improving the commercial efficacy of these biocontrol agents. Entomopathogenic fungi, such as Beauveria bassiana, penetrate the insect cuticle utilizing a plethora of hydrolytic enzymes, including chitinases, which are important virulence factors. Two chitinases (Bbchit1 and Bbchit2) have previously been characterized in B. bassiana, neither of which possesses chitin-binding domains. Here we report the construction and characterization of several B. bassiana hybrid chitinases where the chitinase Bbchit1 was fused to chitin-binding domains derived from plant, bacterial, or insect sources. A hybrid chitinase containing the chitin-binding domain (BmChBD) from the silkworm Bombyx mori chitinase fused to Bbchit1 showed the greatest ability to bind to chitin compared to other hybrid chitinases. This hybrid chitinase gene (Bbchit1-BmChBD) was then placed under the control of a fungal constitutive promoter (gpd-Bbchit1-BmChBD) and transformed into B. bassiana. Insect bioassays showed a 23% reduction in time to death in the transformant compared to the wild-type fungus. This transformant also showed greater virulence than another construct (gpd-Bbchit1) with the same constitutive promoter but lacking the chitin-binding domain. We utilized a strategy where genetic components of the host insect can be incorporated into the fungal pathogen in order to increase host cuticle penetration ability. PMID- 17085714 TI - Gene copy number polymorphisms in an arbuscular mycorrhizal fungal population. AB - Gene copy number polymorphism was studied in a population of the arbuscular mycorrhizal fungus Glomus intraradices by using a quantitative PCR approach on four different genomic regions. Variation in gene copy number was found for a pseudogene and for three ribosomal genes, providing conclusive evidence for a widespread occurrence of macromutational events in the population. PMID- 17085715 TI - Localization of functional polypeptides in bacterial inclusion bodies. AB - Bacterial inclusion bodies, while showing intriguing amyloid-like features, such as a beta-sheet-based intermolecular organization, binding to amyloid-tropic dyes, and origin in a sequence-selective deposition process, hold an important amount of native-like secondary structure and significant amounts of functional polypeptides. The aggregation mechanics supporting the occurrence of both misfolded and properly folded protein is controversial. Single polypeptide chains might contain both misfolded stretches driving aggregation and properly folded protein domains that, if embracing the active site, would account for the biological activities displayed by inclusion bodies. Alternatively, soluble, functional polypeptides could be surface adsorbed by interactions weaker than those driving the formation of the intermolecular beta-sheet architecture. To explore whether the fraction of properly folded active protein is a natural component or rather a mere contaminant of these aggregates, we have explored their localization by image analysis of inclusion bodies formed by green fluorescent protein. Since the fluorescence distribution is not homogeneous and the core of inclusion bodies is particularly rich in active protein forms, such protein species cannot be passively trapped components and their occurrence might be linked to the reconstruction dynamics steadily endured in vivo by such bacterial aggregates. Intriguingly, even functional protein species in inclusion bodies are not excluded from the interface with the solvent, probably because of the porous structure of these particular protein aggregates. PMID- 17085716 TI - Multiple-level regulation of genes for protocatechuate degradation in Acinetobacter baylyi includes cross-regulation. AB - The bacterium Acinetobacter baylyi uses the branched beta-ketoadipate pathway to metabolize aromatic compounds. Here, the multiple-level regulation of expression of the pca-qui operon encoding the enzymes for protocatechuate and quinate degradation was studied. It is shown that both activities of the IclR-type regulator protein PcaU at the structural gene promoter pcaIp, namely protocatechuate-dependent activation of pca-qui operon expression as well as repression in the absence of protocatechuate, can be observed in a different cellular background (Escherichia coli) and therefore are intrinsic to PcaU. The regulation of PcaU expression is demonstrated to be carbon source dependent according to the same pattern as the pca-qui operon. The increase of the pcaU gene copy number leads to a decrease of the basal expression at pcaIp, indicating that the occupancy of the PcaU binding site is well balanced and depends on the concentration of PcaU in the cell. Luciferase is used as a reporter to demonstrate strong repression of pcaIp when benzoate, a substrate of the catechol branch of the pathway, is present in addition to substrates of the protocatechuate branch (cross-regulation). The same repression pattern was observed for promoter pcaUp. Thus, three promoters involved in gene expression of enzymes of the protocatechuate branch (pobAp upstream of pobA, pcaIp, and pcaUp) are strongly repressed in the presence of benzoate. The negative effect of protocatechuate on pobA expression is not based on a direct sensing of the metabolite by PobR, the specific regulator of pobA expression. PMID- 17085717 TI - Units of analysis in accelerated telomere shortening in glycosylphosphatidylinositol (GPI)-negative compared with GPI-positive granulocytes from patients with paroxysmal nocturnal hemoglobinuria (PNH) detected by proaerolysin flow-FISH. PMID- 17085718 TI - HAART- and AIDS-related lymphomas. PMID- 17085719 TI - Analysis of the CD95 ligand gene in 20 children with autoimmune lymphoproliferative syndrome (ALPS). PMID- 17085720 TI - Subtype preference of the BCL6397G/C polymorphism in germinal-center and non germinal-center subtypes of diffuse large B-cell lymphoma. PMID- 17085721 TI - Making sense of apparently conflicting data: stress and immunity in swine and cattle. AB - Stress is generally considered to suppress the immune system and may lead to an increase in the occurrence of disease in the presence of a pathogen. The immune system is ordinarily brought back to a baseline response level after immune challenge through homeostatic processes, in part regulated by the hypothalamic pituitary-axis. Often, findings reported from various studies investigating the effects of stress on the immune system are conflicting and difficult to reconcile into a cohesive and comprehensible set of universally applicable theories. These discrepancies may be partly explained by the types and durations of the stressors, the aspect(s) of immune system measured, genetics, and social status. A particular stressor may enhance cell-mediated immune responses while suppressing humoral responses or vice versa, thus disrupting the balance between these components of the immune system. How farm animals perceive their environment depends not only on traditional environmental stressors (e.g., heat, cold, humidity, pollutants), but also on aspects of their social environment. Dominant animals may have enhanced immune activation, whereas subordinates have suppression of the same immune component in response to the same stressor. This could explain why individual animals within a group respond differently to stressors and disease challenges. A better understanding of the consequences and complex interactions between social and environmental stressors for innate and adaptive immune traits must be developed so we can more fully understand the effects of stress on immunity in livestock. Once these complex relationships are better understood, more effective interventions can be designed to improve animal health and well-being. PMID- 17085722 TI - Combining genetic test information and correlated phenotypic records for breeding value estimation. AB - The use of marker assisted selection in the beef cattle industry to date has involved using traditional EPD in tandem with molecular test information. In the current study, a multiple-trait simulation was carried out to create a beef cattle data set using genetic parameter estimates from the literature to identify the best procedure for combining both sources of information and to assess the added benefit of the procedure. To reach these objectives, the following simulation/ analysis steps were implemented: (1) varying percentages (100, 5, or 0) of available records for the trait of interest, (2) varying percentages (100, 50, 25, or 0) of animals with molecular information, (3) scenarios where the favorable (F) or the unfavorable (U) allele was more frequent, and (4) analysis of the response due to selection over 5 generations. The data sets included 3 correlated traits in which 2 of them, birth weight and postweaning gain, had complete recording and the availability of records for the third trait (marbling score) varied. It was further assumed that molecular information was available for the third trait for a causative gene that explained 10% of the genetic variation. Estimates of Pearson correlations between true and predicted breeding values for marbling score declined as the amount of information declined, and instances in which the molecular information was recorded were always closer to the true values than in the case in which the molecular information was absent. When the U allele was more frequent, rank correlation estimates were increased among top sires, low accuracy sires, and high accuracy sires by approximately 24.9, 12.1, and 4.7% with limited marbling score records and complete genotyping compared with limited marbling score records and no genotyping. Similar results were seen when the F allele was more frequent. When there was a complete absence of recording for the trait of interest, the same trends in correlations were observed and were lower than when the trait of interest was recorded. Jointly considering molecular and phenotypic information showed a greater long-term response compared with tandem selection, showing that discrimination of candidates for selection based solely on molecular information is not optimal. PMID- 17085723 TI - Effect of system of feeding and watering on performance of lactating sows. AB - An experiment was conducted to determine the effects of ad libitum access to feed and water and the option to mix feed and water, all in the same feeder, on the performance of multiparous lactating sows. Feed and water were made available to sows using a self-fed wet/dry (SFWD) or a hand-fed (HF) feed-water system. In the SFWD system, feed and water were dropped into a common trough area of the feeder. The sow determined when and how much of each was dropped. With feed falling onto the flat area of the bottom of the SFWD feeder trough and water falling into the shallow bowl area, and with the 2 areas seamlessly connected, the sow also determined the wetness of the feed consumed. In the HF system, sows were given dry feed twice daily in a J-shaped feeder that was independent of the sow's water source. Sows (n = 114) were assigned to treatments based on parity and genotype. Total feed disappearance per sow during lactation (20 +/- 0.2 d) was greater (P < 0.01) with the SFWD system than with the HF system (120 vs. 110 +/- 4.1 kg, respectively). The SFWD sows had greater (P < 0.01) BW gains during lactation than HF sows (6.2 vs. 0.6 +/- 1.85 kg, respectively). Backfat depth change during lactation did not differ (P = 0.37) between treatments. Likewise, percentage of sows displaying estrus by d 11 post-weaning did not differ (P = 0.51). Piglet weaning BW was greater (P < 0.01) with the SFWD system than with the HF system (6.63 vs. 6.12 +/- 0.22 kg, respectively). Sow average daily water intake and total feed wastage during lactation did not differ (P > 0.66) between treatments. However, sows with the SFWD system wasted less water (P < 0.01) than those with the HF system (15 vs. 232 +/- 12 L, respectively). From a commercial swine production perspective, the difference in waste water volume would result in a significant variation in costs associated with manure storage and distribution. In conclusion, use of a SFWD feed-water system in lactation, which provides sows choices of when to eat, how much to eat, and if dry feed should be mixed with water during consumption, enhances sow appetite, improves litter growth performance, and wastes less water than a HF feed-water system. PMID- 17085724 TI - Board-invited review: recent advances in management of highly stressed, newly received feedlot cattle. AB - Morbidity and mortality from bovine respiratory disease (BRD) and associated losses in performance and carcass merit continue to plague the beef cattle industry. Several viral/bacterial agents are responsible for BRD, and interactions occur among the agents. Viral agents often predispose animals to bacterial infections, and Mannheimia haemolytica is the most frequently isolated organism in cattle with BRD. Laboratory tests are available to characterize organisms causing BRD using easily obtained nasal swab samples. Testing for persistent infection with bovine viral diarrhea virus can be done by a 2-stage technique using PCR and immunohistochemistry. Preconditioning programs that include preweaning viral vaccination programs along with castration could have a significant influence on decreasing BRD in the cattle feeding industry. Metaphylactic antibiotic programs continue to be effective; however, antibiotic resistance is a public concern, and additional management options (e.g., direct fed microbials or other compounds with antimicrobial properties) deserve attention. Diets with an increased energy concentration achieved by decreasing the dietary roughage concentration may slightly increase the rate of BRD morbidity; however, these diets also increase ADG, DMI, and G:F compared with lower-energy, greater-roughage diets. The extent to which performance and BRD morbidity are affected by dietary protein concentration needs further study, but low and high protein concentrations should probably be avoided. Several trace minerals (e.g., Cu, Se, and Zn) affect immune function, but the effects of supplementation on performance and immune function in model challenge systems and in field studies are equivocal. Adding vitamin E to receiving diets at pharmacological levels (e.g., >1,000 IU x animal(-1) x day(-1)) seems beneficial for decreasing BRD morbidity, but it has little effect on performance. Given the limited ability to consistently modify immune function and BRD morbidity through dietary manipulations, we recommend that the diets for newly received cattle be formulated to adjust nutrient concentrations for low feed intake and to provide optimal performance during the receiving period. PMID- 17085725 TI - Seminal fluid signaling in the female reproductive tract: lessons from rodents and pigs. AB - Seminal fluid contains potent signaling agents that influence female reproductive physiology to improve the chances of conception and pregnancy success. Cytokines and prostaglandins synthesized in the male accessory glands are transferred to the female at insemination, where they bind to receptors on target cells in the cervix and uterus, activating changes in gene expression that lead to modifications in structure and function of the female tissues. The consequences are increased sperm survival and fertilization rates, conditioning of the female immune response to tolerate semen and the conceptus, and molecular and cellular changes in the endometrium that facilitate embryo development and implantation. Male-female tract signaling occurs in rodents, livestock animals, and all other mammals examined thus far, including humans. In mice, the key signaling moieties in seminal plasma are identified as members of the transforming growth factor beta family. Recent studies indicate a similar signaling function for boar factors in the pig, whereby the sperm and plasma fractions of seminal fluid appear to synergize in activating an inflammatory response and downstream changes in the female tract after insemination. Seminal plasma elicits endometrial changes, with induction of proinflammatory cytokines and cyclooxygenase-2, causing recruitment of macrophages and dendritic cells. Sperm contribute by interacting with seminal plasma factors to modulate neutrophil influx into the luminal cavity. The cascade of changes in local leukocyte populations and cytokine synthesis persists throughout the preimplantation period. Exposure to seminal fluid alters the dynamics of preimplantation embryo development, with an increase in the number of fertilized oocytes attaining the viable blastocyst stage. There is also evidence that seminal factors influence the timing of ovulation, corpus luteum development, and progesterone synthesis. Insight into the molecular basis of seminal fluid signaling in the female reproductive tract may inform new interventions and management practices to ensure maximal fertility and reduce embryo mortality in pigs and, potentially, other livestock species. PMID- 17085726 TI - Shiga toxin-producing Escherichia coli: an overview. AB - The objective of this review is to highlight the importance of cattle in human disease due to Shiga toxin-producing Escherichia coli (STEC) and to discuss features of STEC that are important in human disease. Healthy dairy and beef cattle are a major reservoir of a diverse group of STEC that infects humans through contamination of food and water, as well as through direct contact. Infection of humans by STEC may result in combinations of watery diarrhea, bloody diarrhea, and hemolytic uremic syndrome. Systems of serotyping, subtyping, and virulence typing of STEC are used to aid in epidemiology, diagnosis, and pathogenesis studies. Severe disease and outbreaks of disease are most commonly due to serotype O157:H7, which, like most other highly pathogenic STEC, colonize the large intestine by means of a characteristic attaching and effacing lesion. This lesion is induced by a bacterial type III secretion system that injects effector proteins into the intestinal epithelial cell, resulting in profound changes in the architecture and metabolism of the host cell and intimate adherence of the bacteria. Severe disease in the form of bloody diarrhea and the hemolytic uremic syndrome is attributable to Shiga toxin (Stx), which exists as 2 major types, Stx1 and Stx2. The stx genes are encoded on temperate bacteriophages in the chromosome of the bacteria, and production and release of the toxin are highly dependent on induction of the phages. Regulation of the genes involved in induction of the attaching and effacing lesion, and production of Stx is complex. In addition to these genes that are clearly implicated in virulence, there are several putative virulence factors. A major public health goal is to prevent STEC induced disease in humans. Studies aimed at understanding factors that affect carriage and shedding of STEC by cattle and factors that contribute to development of disease in humans are considered to be important in achieving this objective. PMID- 17085727 TI - Acclimation to high ambient temperature in Large White and Caribbean Creole growing pigs. AB - The effect of breed [Creole (CR) vs. Large White (LW)] on performance and physiological responses during acclimation to high ambient temperature was studied in 2 experiments involving 24 (12/breed) growing pigs each. Pigs were exposed to 24 degrees C for 10 d (d -10 to -1) and thereafter to a constant temperature of 31 degrees C for 16 d (d 1 to d 16) in Exp. 1 and for 20 d (d 1 to d 20) in Exp. 2. For both experiments, the temperature change was achieved over 4 h on d 0. The first experiment began at 105 d of age, and the average BW of CR and LW pigs was 36.6 +/- 2.5 kg and 51.7 +/- 3.0 kg, respectively. The second experiment was designed to compare both breeds at a similar BW (about 52 kg on d 0). Pigs were individually housed and given ad libitum access to feed. At 24 degrees C, ADG was lower (P < 0.01) in CR than in LW (602 vs. 913 g/d and 605 vs. 862 g/d in Exp. 1 and 2, respectively), but the ADFI was not affected by breed (190 and 221 g x d(-1) x kg(-0.60) in Exp. 1 and 2, respectively). Short-term thermoregulatory responses during the 4-h transition from 24 to 31 degrees C (d 0) were analyzed according to a linear plateau model to determine the break point temperature, above which rectal temperature (RT), cutaneous temperature (CT), and respiratory rate (RR) began to change. The CT increased linearly with temperature increase (0.22 degrees C/ degrees C) and was less (P < 0.05) in CR than in LW (by -0.3 degrees C on average). In both experiments, the break point temperature for RT was not affected by breed (27.6 degrees C on average), whereas for RR it was greater (P < 0.05) in CR than in LW (27.5 vs. 25.5 degrees C, P < 0.01). On average, ADFI declined by about 50 g x d(-1) x kg(-0.60) from d -1 to d 1 (P < 0.01), and thereafter at 31 degrees C, it gradually increased (23 g x d(-1) x kg( 0.60); P < 0.05), suggesting an acclimation to high exposure. This response was not influenced by breed. After the day that marked the beginning of the acclimation response (i.e., the threshold day), RR, CT, and RT declined over the duration of exposure to 31 degrees C (P < 0.05) in both experiments. During this period, RT and CT were less in CR than in LW pigs (39.6 vs. 39.9 degrees C and 37.9 vs. 38.2 degrees C, respectively; P < 0.05), whereas RR was not affected by breed. The threshold day at which RT began to decline was less in CR than in LW pigs (0.18 vs. 1.17 d and 0.39 vs. 0.93 d in Exp. 1 and 2, respectively; P < 0.05). In conclusion, this study suggests that short- and long-term physiological reactions during heat acclimation differed when CR and LW pigs were compared at the same age or BW. PMID- 17085728 TI - Technical note: Use of marker-based relationships with multiple-trait derivative free restricted maximal likelihood. AB - The widespread use of the set of multiple-trait derivative-free REML programs for prediction of breeding values and estimation of variance components has led to significant improvement in traits of economic importance. The initial version of this software package, however, was generally limited to pedigree-based relationships. With continued advances in genomic research and the increased availability of genotyping, relationships based on molecular markers are obtainable and desirable. The addition of a new program to the set of multiple trait derivative-free REML programs is described that allows users the flexibility to calculate relationships using standard pedigree files or an arbitrary relationship matrix based on genetic marker information. The strategy behind this modification and its design is described. An application is illustrated in a QTL association study for canine hip dysplasia. PMID- 17085729 TI - Relationship between bone strength and dual-energy X-ray absorptiometry measurements in pigs. AB - Computed tomography and a 3-point bending test were performed on the metacarpal bones of adult production pigs to test the hypothesis that bone strength is strongly correlated with areal bone mineral density (BMD) in this population. The aim of the study was to subject material from adult production pigs grouped by BMD to 3-point bending, to test this hypothesis and determine any correlations. In all, 168 individual computed tomography scans and mechanical tests were performed on the collected material. For evaluation purposes, the material was divided into the categories low, medium, and high BMD (<1, 1 to 1.4, and >1.4 g/cm(2), respectively). The results showed a difference in the maximum load, in the stress at maximum load, and stiffness among each BMD group (P < 0.001) and in elastic modulus between the low BMD group and the 2 other groups (P < 0.001). A correlation between both intrinsic and extrinsic measures of bone strength and BMD was thus demonstrated. The projected change in each of the variables reported, for a 0.1 g/cm(2) alteration in BMD (within the BMD range evaluated in this study), is as follows: maximum load, 708 N; stress at maximum load, 50 N/mm(2); stiffness, 391.6 N/mm; and elastic modulus, 108 N/mm(2) (P < 0.001). The results confirm the relationship between BMD and bone strength and indicate that BMD screening can be used in fracture risk assessments in production pigs. PMID- 17085730 TI - Effect of supplemental iron on finishing swine performance, carcass characteristics, and pork quality during retail display. AB - Crossbred pigs (n = 185) were used to test the effects of dietary Fe supplementation on performance and carcass characteristics of growing-finishing swine. Pigs were blocked by BW, allotted to pens (5 to 6 pigs/pen), and pens (5 pens/block) were allotted randomly to either negative control (NC) corn-soybean meal grower and finisher diets devoid of Fe in the mineral premix, positive control (PC) corn-soybean meal grower and finisher diets with Fe included in the mineral premix, or the PC diets supplemented with 50, 100, or 150 ppm Fe from Availa-Fe (an Fe-AA complex). When the lightest block averaged 118.2 kg, the pigs were slaughtered, and bone-in pork loins were collected during fabrication for pork quality data. During the grower-I phase, there was a tendency for supplemental Fe to reduce ADG linearly (P = 0.10), whereas in the grower-II phase, supplemental Fe tended to increase ADG linearly (P = 0.10). Even though pigs fed NC had greater G:F during the finisher-I phase (P < 0.05) and across the entire trial (P = 0.07), live performance did not (P > or = 0.13) differ among dietary treatments. There were linear increases in 10th-rib fat depth (P = 0.08) and calculated fat-free lean yield (P = 0.06); otherwise, dietary Fe did not (P > 0.19) affect pork carcass muscling or fatness. Moreover, LM concentrations of total, heme, and nonheme Fe were similar (P > 0.23) among treatments. A randomly selected subset of loins from each treatment was further fabricated into 2.5-cm thick LM chops, placed on styrofoam trays, overwrapped with polyvinyl chloride film, and placed in coffin-chest display cases (2.6 degrees C) under continuous fluorescent lighting (1,600 lx) for 7 d. During display, chops from NC-fed pigs and pigs fed the diets supplemented with 100 ppm Fe tended to have a more vivid (higher chroma value; P = 0.07), redder (higher a* value; P = 0.09) color than LM chops of pigs fed 50 ppm of supplemental Fe. Moreover, greater (P < 0.01) redness:yellowness ratios in chops from pigs supplemented with 100 ppm Fe indicated a more red color than chops from PC-fed pigs or pigs fed diets supplemented with 50 ppm Fe. In conclusion, however, increasing dietary Fe had no appreciable effects on performance, carcass, or LM characteristics, suggesting that current dietary Fe recommendations are sufficient for optimal growth performance, pork carcass composition, and pork quality. PMID- 17085731 TI - Anthelmintic and nutritional effects of heather supplementation on Cashmere goats grazing perennial ryegrass-white clover pastures. AB - To investigate anthelmintic and nutritional effects of heather supplementation in goats grazing perennial ryegrass-white clover pastures, 40 dry Cashmere goats were randomly assigned to 4 treatments in a 2 x 2 factorial arrangement: 2 grazing management treatments (supplementation with heather vs. nonsupplementation) and 2 anthelmintic treatments (treatment vs. nontreatment). Goats grazed continuously from May to September 2004. At the end of the grazing period, the number of dead goats due to gastrointestinal parasitism was 1 in the group supplemented with heather and dosed with anthelmintic, 4 in the group that received neither supplementation nor anthelmintic, and 0 in the other 2 groups. For goats that did not receive anthelmintic treatment, the percentage of heather in the diet was negatively correlated with fecal egg count in August (r = -0.59, P < 0.05) and September (r = -0.49, P < 0.1) and positively correlated (r = 0.54, P < 0.05) with BW changes during the grazing season. Therefore, the correlation coefficient between BW change and fecal egg count was negative (r = -0.62, P < 0.05). Rumen ammonia concentrations were always lower in supplemented goats (P < 0.05). However, VFA concentrations were greater in goats consuming heather (58.9 vs. 50.9 mmol/L), which suggests that ruminal fermentation was not adversely affected by consumption of tannins. Heather availability in the vegetation might represent a valuable opportunity and sustainable method to control gastrointestinal nematode infections in a goat production system based on grazing perennial ryegrass-white clover pastures. PMID- 17085732 TI - Stimulation of estrous behavior in grazing female goats by continuous or discontinuous exposure to males. AB - Two experiments were conducted during the anestrous period to determine: (1) whether males rendered sexually active by exposure to artificial long days stimulate estrous activity of female goats under grazed conditions (Exp. 1); and (2) whether continuous presence of the buck is necessary to stimulate this estrous activity (Exp. 2). In Exp. 1, 2 groups of females (n = 20/group), one in confinement and another under grazing conditions, were exposed to 4 bucks subjected to natural photoperiod (2 males/group). Two other groups of females (n = 20/group), in confinement or grazing, were exposed to 4 males treated with artificial long days (2 males/group). All groups were exposed to males for 15 d. The percentage of does detected in estrus during these 15 d was greater (P < 0.001) in the 2 groups exposed to males sexually prepared by long days (confined, 95%; grazed, 90%) than in groups exposed to males in natural photoperiod (confined, 15%; grazed, 45%). Does in Exp. 2 were allowed to graze and were exposed continuously (n = 26) or discontinuously (from 1700 to 0900; n = 26) for 18 d to males that had been stimulated to enter the breeding season by exposure to long days. The proportion of does that displayed estrous behavior in 18 d did not differ (P = 0.55) between groups (96.2 and 92.3% for continuous and discontinuous groups, respectively). The results indicate that anestrous goats managed under grazing conditions can be stimulated to express estrus by joining with males previously exposed to artificial long days. Continuous presence of the male is not necessary for this male effect. PMID- 17085733 TI - Conception rates to artificial insemination in primiparous, suckled cows exposed to the biostimulatory effect of bulls before and during a gonadotropin-releasing hormone-based estrus synchronization protocol. AB - The objective of these studies was to evaluate whether exposing primiparous, suckled beef cows to the biostimulatory effect of bulls alters breeding performance associated with an estrus synchronization protocol that included GnRH followed 7 d later by PGF(2alpha) and fixed-time AI (TAI). This was a composite analysis of 3 experiments that evaluated (1) the effects of bull exposure at different days after calving (yr 1); (2) the biostimulatory effects of bull excretory products (yr 2); and (3) the biostimulatory effects of familiar and unfamiliar bulls (yr 3) on the resumption of ovarian cycling activity. In all studies, cows were exposed (biostimulated; n = 94) or not exposed (nonbiostimulated; n = 67) to bulls or excretory products of bulls for at least 60 d before the beginning of the estrus synchronization protocol. Average calving day did not differ among years and was 52 +/- 5 d. Year did not affect the proportions of biostimulated and nonbiostimulated cows that were cycling at the beginning of the estrus synchronization protocol; however, a greater (P < 0.001) proportion of biostimulated than nonbiostimulated cows were cycling at this time. In each year, cows were given GnRH followed by PGF(2alpha) 7 d later. Cows were observed for estrus twice daily (am and pm) after PGF(2alpha). Cows that exhibited estrus before 54, 60, and 64 h after PGF(2alpha) were inseminated by AI 12 h later in yr 1, 2, and 3, respectively. Cows that failed to show estrus were given GnRH and TAI at 62, 72, and 72 h after PGF(2alpha) in yr 1, 2, and 3, respectively. Conception rates were determined by transrectal ultrasonography 35 d after TAI in each year. The percentages of cows that exhibited estrus after PGF(2alpha) and before TAI, the interval from PGF(2alpha) to estrus, and the percentages of cows inseminated 12 h after estrus or at TAI did not differ between biostimulated and nonbiostimulated cows and were 51%, 54.7 +/- 7.3 h, 35%, and 65%, respectively. Conception rates for cows bred by AI 12 h after estrus did not differ between biostimulated and nonbiostimulated cows; however, the TAI conception rate was greater (P < 0.05) for biostimulated cows (57.6%) than for nonbiostimulated cows (35.6%). We conclude that TAI conception rates in an estrus synchronization protocol that includes GnRH followed 7 d later by PGF(2alpha) may be improved by the biostimulatory effect of bulls in postpartum, primiparous cows. PMID- 17085734 TI - Weaning weight inheritance in environments classified by maternal body weight change. AB - In good environments, cow intake is sufficient for their own growth and for milk production to support their calf. In poor environments, cows lose BW or may reduce milk supply to maintain themselves. Heritability for direct genetic and maternal components of weaning weight as well as the correlations between these components might be expected to vary according to these circumstances. The purpose of this study was to estimate heritability and genetic correlations for the direct genetic and maternal components of weaning weight classified in 2 environments according to maternal BW gain and to identify whether a single heritability estimate is appropriate for the differing environments experienced by cows from year to year. Data used in this analysis was obtained from the Red Angus Association of America and consisted of 96,064 cow BW observations and 27,534 calf weaning weight observations. A dam's change in BW from one year to the next was used to classify each calf's weaning weight into 1 of 2 environmental groups, those being good or poor. Best linear unbiased estimates of the change in cow BW with age were obtained from analysis of cow BW using a repeatability model. If the phenotypic change in cow BW exceeded this average BW change, the calf's weaning weight associated with the end of this time frame was classified as having been observed in a good environment. If not, the calf's corresponding weaning weight was classified as having occurred in a poorer than average environment. Heritability estimates of 0.24 +/- 0.03, 0.24 +/- 0.03, 0.13 +/- 0.02, and 0.14 +/- 0.02 were obtained for weaning weight good direct, poor direct, good maternal, and poor maternal, respectively. Correlations between direct genetic and maternal weaning weight components in the good and poor environments were -0.47 +/- 0.08 and -0.20 +/- 0.09, respectively. These variance components are not sufficiently distinct to warrant accounting for dam nutritional environment in national cattle evaluation. PMID- 17085735 TI - Effects of dam nutrition on growth and reproductive performance of heifer calves. AB - A 3-yr study was conducted with heifers (n = 170) whose dams were used in a 2 x 2 factorial arrangement of treatments to determine the effects of late gestation (LG) or early lactation (EL) dam nutrition on subsequent heifer growth and reproduction. In LG, cows received 0.45 kg/d of a 42% CP supplement (PS) or no supplement (NS) while grazing dormant Sandhills range. During EL, cows from each late gestational treatment were fed cool-season grass hay or grazed sub-irrigated meadow. Cows were managed as a single herd for the remainder of the year. Birth date and birth weight of heifer calves were not affected (P > 0.10) by dam nutrition. Meadow grazing and PS increased (P = 0.02; P = 0.07) heifer 205-d BW vs. feeding hay and NS, respectively. Weight at prebreeding and pregnancy diagnosis were greater (P < 0.04) for heifers from PS dams but were unaffected by EL nutrition (P > 0.10). There was no effect (P > 0.10) of LG or EL dam nutrition on age at puberty or the percentage of heifers cycling before breeding. There was no difference (P > 0.10) in pregnancy rates due to EL treatment. Pregnancy rates were greater (P = 0.05) for heifers from PS dams, and a greater proportion (P = 0.005) of heifers from PS dams calved in the first 21 d of the heifers' first calving season. Nutrition of the dams did not influence (P < 0.10) heifers' average calving date, calving difficulty, and calf birth weight during the initial calving season. Weight at the beginning of the second breeding season was greater (P = 0.005) for heifers from PS dams but was not affected by maternal nutrition during EL (P > 0.10). Dam nutrition did not affect (P > 0.10) heifer ADG or G:F ratio. Heifers from PS dams had greater DMI (P = 0.09) and residual feed intake (P = 0.07) than heifers from NS cows if their dams were fed hay during EL but not if their dams grazed meadows. Heifers born to PS cows were heavier at weaning, prebreeding, first pregnancy diagnosis, and before their second breeding season. Heifers from cows that grazed meadows during EL were heavier at weaning but not postweaning. Despite similar ages at puberty and similar proportions of heifers cycling before the breeding season, a greater proportion of heifers from PS dams calved in the first 21 d of the heifers' first calving season, and pregnancy rates were greater compared with heifers from NS dams. Collectively, these results provide evidence of a fetal programming effect on heifer postweaning BW and fertility. PMID- 17085736 TI - The science of surge. PMID- 17085737 TI - Daily patient flow is not surge: "management is prediction". PMID- 17085738 TI - Understanding surge capacity: essential elements. AB - As economic forces have reduced immediately available resources, the need to surge to meet patient care needs that exceed expectations has become an increasing challenge to the health care community. The potential patient care needs projected by pandemic influenza and bioterrorism catapulted medical surge to a critical capability in the list of national priorities, making it front-page news. Proposals to improve surge capacity are abundant; however, surge capacity is poorly defined and there is little evidence-based comprehensive planning. There are no validated measures of effectiveness to assess the efficacy of interventions. Before implementing programs and processes to manage surge capacity, it is imperative to validate assumptions and define the underlying components of surge. The functional components of health care and what is needed to rapidly increase capacity must be identified by all involved. Appropriate resources must be put into place to support planning factors. Using well-grounded scientific principles, the health care community can develop comprehensive programs to prioritize activities and link the necessary resources. Building seamless surge capacity will minimize loss and optimize outcomes regardless of the degree to which patient care needs exceed capability. PMID- 17085739 TI - Surge capacity for health care systems: early detection, methodologies, and process. AB - Excessive demand on hospital services from large-scale emergencies is something that every emergency department health care provider and hospital administrator knows could happen at any time. Nowhere in this country have we recently faced a disaster of the magnitude of concern we now face involving agents of mass destruction or social disruption, especially those in the area of infectious diseases and radiological materials. The war on terrorism is not a conventional war, and terrorists may use any means of convenience to carry out their objectives in an unpredictable time line. Have we adequately prepared for the potentially excessive surge in demand for medical services that a large-scale event could bring to our medical care system? Are our emergency departments ready for such events? Surveillance systems, such as BioWatch, BioSense, the National Biosurveillance Integration System, and the countermeasure program BioShield, offer hope that we will be able to meet these new challenges. PMID- 17085740 TI - Individual-based computational modeling of smallpox epidemic control strategies. AB - In response to concerns about possible bioterrorism, the authors developed an individual-based (or "agent-based") computational model of smallpox epidemic transmission and control. The model explicitly represents an "artificial society" of individual human beings, each implemented as a distinct object, or data structure in a computer program. These agents interact locally with one another in code-represented social units such as homes, workplaces, schools, and hospitals. Over many iterations, these microinteractions generate large-scale macroscopic phenomena of fundamental interest such as the course of an epidemic in space and time. Model variables (incubation periods, clinical disease expression, contagiousness, and physical mobility) were assigned following realistic values agreed on by an advisory group of experts on smallpox. Eight response scenarios were evaluated at two epidemic scales, one being an introduction of ten smallpox cases into a 6,000-person town and the other an introduction of 500 smallpox cases into a 50,000-person town. The modeling exercise showed that contact tracing and vaccination of household, workplace, and school contacts, along with prompt reactive vaccination of hospital workers and isolation of diagnosed cases, could contain smallpox at both epidemic scales examined. PMID- 17085741 TI - Differentiating large-scale surge versus daily surge. AB - This breakout session at the Academic Emergency Medicine 2006 Consensus Conference examined how baseline overcrowding impedes the ability of emergency departments to respond to sudden, unexpected surges in demand for patient care. Differences between daily and catastrophic surge were discussed, and the need to invoke a hospital-wide response to surge was explored. PMID- 17085743 TI - Antibodies to Toxoplasma gondii in patients with schizophrenia: a meta-analysis. AB - Recent studies have linked infectious agents to schizophrenia. The largest number of studies has involved the analysis of Toxoplasma gondii; these studies were subjected to a meta-analysis. Published articles and abstracts were identified by searches of MEDLINE, Ovid, and Google Scholar; by a search of Chinese publications; through letters to researchers; and by visiting China. Published and unpublished controlled studies that used serological methods for measuring T. gondii antibodies to assess inpatients and/or outpatients diagnosed with schizophrenia were selected for analysis, and source documents were translated as needed. Forty-two studies carried out in 17 countries over 5 decades were identified; 23 of these (6 unpublished) met selection criteria. The combined odds ratio (OR) was 2.73 (95% confidence interval, 2.10 to 3.60; chi-square with 1 df 263; P < .000001). Seven studies that included only patients with first-episode schizophrenia (OR 2.54) did not differ significantly from 16 studies that included patients in all clinical phases (OR 2.79). The results suggest that individuals with schizophrenia have an increased prevalence of antibodies to T. gondii. This association is consistent with other epidemiological studies as well as with animal studies. Although the OR of 2.73 is modest, it exceeds that for genetic or other environmental factors identified to date and suggests that Toxoplasma is in some way associated with a large number of cases of schizophrenia. If an etiological association can be proven, it would have implications for the design of measures for the prevention and treatment of this disease. PMID- 17085744 TI - Re: "estimating the proportion of disease due to classes of sufficient causes". PMID- 17085745 TI - Insights into latent class analysis of diagnostic test performance. AB - Latent class analysis is used to assess diagnostic test accuracy when a gold standard assessment of disease is not available but results of multiple imperfect tests are. We consider the simplest setting, where 3 tests are observed and conditional independence (CI) is assumed. Closed-form expressions for maximum likelihood parameter estimates are derived. They show explicitly how observed 2- and 3-way associations between test results are used to infer disease prevalence and test true- and false-positive rates. Although interesting and reasonable under CI, the estimators clearly have no basis when it fails. Intuition for bias induced by conditional dependence follows from the analytic expressions. Further intuition derives from an Expectation Maximization (EM) approach to calculating the estimates. We discuss implications of our results and related work for settings where more than 3 tests are available. We conclude that careful justification of assumptions about the dependence between tests in diseased and nondiseased subjects is necessary in order to ensure unbiased estimates of prevalence and test operating characteristics and to provide these estimates clinical interpretations. Such justification must be based in part on a clear clinical definition of disease and biological knowledge about mechanisms giving rise to test results. PMID- 17085746 TI - Peak expiratory flow monitoring in older patients with asthma: An aid to management? PMID- 17085747 TI - Intrinsic differences of the airway epithelium in childhood allergic asthma. PMID- 17085748 TI - Schedule or dosage? The need to perfect intermittent regimens for tuberculosis. PMID- 17085749 TI - Is maximal lung recruitment worth it? PMID- 17085750 TI - The role of central vasopressin receptors in the modulation of autonomic cardiovascular controls: a spectral analysis study. AB - Although it has been suggested that vasopressin (VP) acts within the central nervous system to modulate autonomic cardiovascular controls, the mechanisms involved are not understood. Using nonpeptide, selective V(1a), V(1b), and V(2) antagonists, in conscious rats, we assessed the roles of central VP receptors, under basal conditions, after the central application of exogenous VP, and after immobilization, on cardiovascular short-term variability. Equidistant sampling of blood pressure (BP) and heart rate (HR) at 20 Hz allowed direct spectral analysis in very-low frequency (VLF-BP), low-frequency (LF-BP), and high-frequency (HF-BP) blood pressure domains. The effect of VP antagonists and of exogenous VP on body temperature (T(b)) was also investigated. Under basal conditions, V(1a) antagonist increased HF-BP and T(b), and this was prevented by metamizol. V(1b) antagonist enhanced HF-BP without affecting T(b), and V(2) antagonist increased VLF-BP variability which could be prevented by quinapril. Immobilization increased BP, LF-BP, HF-BP, and HF-HR variability. V(1a) antagonist prevented BP and HR variability changes induced by immobilization and potentiated tachycardia. V(1b) antagonist prevented BP but not HR variability changes, whereas V(2) antagonist had no effect. Exogenous VP increased systolic arterial pressure (SAP) and HF-SAP variability, and this was prevented by V(1a) and V(1b) but not V(2) antagonist pretreatment. Our results suggest that, under basal conditions, VP, by stimulation of V(1a), V(1b), and cognate V(2) receptors, buffers BP variability, mostly due to thermoregulation. Immobilization and exogenous VP, by stimulation of V(1a) or V(1b), but not V(2) receptors, increases BP variability, revealing cardiorespiratory adjustment to stress and respiratory stimulation, respectively. PMID- 17085751 TI - Effects of brief cutaneous JP-8 jet fuel exposures on time course of gene expression in the epidermis. AB - The jet fuel jet propulsion fuel 8 (JP-8) has been shown to cause an inflammatory response in the skin, which is characterized histologically by erythema, edema, and hyperplasia. Studies in laboratory animal skin and cultured keratinocytes have identified a variety of changes in protein levels related to inflammation, oxidative damage, apoptosis, and cellular growth. Most of these studies have focused on prolonged exposures and subsequent effects. In an attempt to understand the earliest responses of the skin to JP-8, we have investigated changes in gene expression in the epidermis for up to 8 h after a 1-h cutaneous exposure in rats. After exposure, we separated the epidermis from the rest of the skin with a cryotome and isolated total mRNA. Gene expression was studied with microarray techniques, and changes from sham treatments were analyzed and characterized. We found consistent twofold increases in gene expression of 27 transcripts at 1, 4, and 8 h after the beginning of the 1-h exposure that were related primarily to structural proteins, cell signaling, inflammatory mediators, growth factors, and enzymes. Analysis of pathways changed showed that several signaling pathways were increased at 1 h and that the most significant changes at 8 h were in metabolic pathways, many of which were downregulated. These results confirm and expand many of the previous molecular studies with JP-8. Based on the 1-h changes in gene expression, we hypothesize that the trigger of the JP-8 induced, epidermal stress response is a physical disruption of osmotic, oxidative, and membrane stability which activates gene expression in the signaling pathways and results in the inflammatory, apoptotic, and growth responses that have been previously identified. PMID- 17085752 TI - Innovative application of a multidimensional item response model in assessing the influence of social desirability on the pseudo-relationship between self-efficacy and behavior. AB - This study examined multidimensional item response theory (MIRT) modeling to assess social desirability (SocD) influences on self-reported physical activity self-efficacy (PASE) and fruit and vegetable self-efficacy (FVSE). The observed sample included 473 Houston-area adolescent males (10-14 years). SocD (nine items), PASE (19 items) and FVSE (21 items) were measured with previously validated self-report instruments containing Likert-type responses. Physical activity was objectively measured using the Computer Science Application Incorporated/Manufacturing Technology Incorporated (CSA/MTI) accelerometer. Total fruit, juice and vegetable consumption was measured with a food frequency questionnaire. Correlations between self-efficacy and behaviors were minimal, regardless of controlling for SocD. However, in a simulated sample derived to demonstrate the utility of MIRT when relationships exist, the pseudo relationships between self-efficacy and behaviors were substantially weaker after controlling for SocD. MIRT provided disattenuated correlations between SocD and self-efficacy, thereby providing more precise estimates of the real influence of SocD on the relationship between self-efficacy and behavior. However, as shown in the observed sample, more research is needed to understand the influence of SocD on the relationship between self-efficacy and behaviors for different populations and for different degrees of SocD response bias. PMID- 17085753 TI - Aluminium toxicity in plants: internalization of aluminium into cells of the transition zone in Arabidopsis root apices related to changes in plasma membrane potential, endosomal behaviour, and nitric oxide production. AB - The extent of aluminium internalization during the recovery from aluminium stress in living roots of Arabidopsis thaliana was studied by non-invasive in vivo microscopy in real time. Aluminium exposure caused rapid depolarization of the plasma membrane. The extent of depolarization depends on the developmental state of the root cells; it was much more extensive in cells of the distal than in the proximal portion of the transition zone. Also full recovery of the membrane potential after removal of external aluminium was slower in cells of the distal transition zone than of its proximal part. Using morin, a vital marker dye for aluminium, and FM4-64, a marker for endosomal/vacuolar membranes, an extensive aluminium internalization was recorded during the recovery phase into endosomal/vacuolar compartments in the most aluminium-sensitive cells. Interestingly, aluminium interfered with FM4-64 internalization and inhibited the formation of brefeldin A-induced compartments in these cells. By contrast, there was no detectable uptake of aluminium into cells of the proximal part of the transition zone and the whole elongation region. Moreover, cells of the distal portion of the transition zone emitted large amounts of nitric oxide (NO) and this was blocked by aluminium treatment. These data suggest that aluminium internalization is related to the most sensitive status of the distal portion of the transition zone towards aluminium. Aluminium in these root cells has impact on endosomes and NO production. PMID- 17085754 TI - A permeation-diffusion-reaction model of gas transport in cellular tissue of plant materials. AB - Gas transport in fruit tissue is governed by both diffusion and permeation. The latter phenomenon is caused by overall pressure gradients which may develop due to the large difference in O(2) and CO(2) diffusivity during controlled atmosphere storage of the fruit. A measurement set-up for tissue permeation based on unsteady-state gas exchange was developed. The gas permeability of pear tissue was determined based on an analytical gas transport model. The overall gas transport in pear tissue samples was validated using a finite element model describing simultaneous O(2), CO(2), and N(2) gas transport, taking into account O(2) consumption and CO(2) production due to respiration. The results showed that the model described the experimentally determined permeability of N(2) very well. The average experimentally determined values for permeation of skin, cortex samples, and the vascular bundle samples were (2.17+/-1.71)x10(-19) m(2), (2.35+/ 1.96)x10(-19) m(2), and (4.51+/-3.12)x10(-17) m(2), respectively. The permeation diffusion-reaction model can be applied to study gas transport in intact pears in relation to product quality. PMID- 17085755 TI - Ethylene could influence ferric reductase, iron transporter, and H+-ATPase gene expression by affecting FER (or FER-like) gene activity. AB - In previous works, it has been shown, by using ethylene inhibitors and precursors, that ethylene could participate in the regulation of the enhanced ferric reductase activity of Fe-deficient Strategy I plants. However, it was not known whether ethylene regulates the ferric reductase gene expression or other aspects related to this activity. This paper is a study of the effects of ethylene inhibitors and precursors on the expression of the genes encoding the ferric reductases and iron transporters of Arabidopsis thaliana (FRO2 and IRT1) and Lycopersicon esculentum (=Solanum lycopersicum) (FRO1 and IRT1) plants. The effects of ethylene inhibitors and precursors on the activity of the iron reductase and the iron transporter have been examined in parallel. Also studied were the effects of ethylene inhibitors and precursors on the expression of the H(+)-ATPase genes of cucumber (CsHA1 and CsHA2) and the transcription factor genes of tomato (LeFER) and Arabidopsis (AtFRU or AtFIT1, an LeFER homologue) that regulate ferric reductase, iron transporter, and H(+)-ATPse activity. The results obtained suggest that ethylene participates in the regulation of ferric reductase, the iron transporter, and H(+)-ATPase gene expression by affecting the FER (or FER-like) levels. PMID- 17085756 TI - Further quantification of the role of internal unstirred layers during the measurement of transport coefficients in giant internodes of Chara by a new stop flow technique. AB - A new stop-flow technique was employed to quantify the impact of internal unstirred layers on the measurement of the solute permeability coefficient (P(s)) across the plasma membrane of internodes of the giant-celled alga Chara corallina using a cell pressure probe. During permeation experiments with rapidly permeating solutes (acetone, 2-propanol, and dimethylformamide), the solute concentration inside the cell was estimated and the external medium was adjusted to stop solute transport across the membrane, after which responses in turgor were measured. This allowed estimation of the solute concentration right at the membrane. Stop-flow experiments were also simulated with a computer. Both the stop-flow experiments and simulations provided quantitative data about internal concentration gradients and the contribution of unstirred layers to overall measured values of P(meas)(s) for the three solutes. The stop-flow experimental results agreed with stop-flow simulations assuming that solutes diffused into a completely stagnant cell interior. The effects of internal unstirred layers on the underestimation of membrane P(s) declined with decreasing P(s). They were no bigger than 37% in the presence of the most rapidly permeating solute, acetone (P(meas)(s) =4.2 x 10(-6) m s(-1)), and 14% for the less rapidly permeating dimethylformamide (P(meas)(s) =1.6x10(-6) m s(-1)). It is concluded that, even in the case of rapidly permeating solutes such as isotopic water and, even when making pessimistic assumptions about the internal mixing of solutes, an upper limit for the underestimation of P(s) due to internal unstirred layers was 37%. The data are discussed in terms of recent theoretical estimates of the effect of internal unstirred layers and in terms of some recent criticism of cell pressure probe measurements of water and solute transport coefficients. The current stop flow data are in line with earlier estimations of the role of unstirred layers in the literature on cell water relations. PMID- 17085757 TI - Predicting lewy body pathology in a community-based sample with clinical diagnosis of Alzheimer's disease. AB - Accurate antemortem prediction of Lewy body pathology in patients with dementia is problematic. This study generates a model that better predicts Lewy body pathology in community-based patients with clinical Alzheimer's disease. Lewy body pathology was detected in 80 of 152 participants (52.6%) with an initial diagnosis of probable Alzheimer's disease. In a stepwise logistic regression model, female gender, lower education, being married, bradykinesia, hallucinations, and absence of irritability predicted the greatest likelihood of Lewy body pathology. The predictive model correctly diagnosed Lewy body pathology with an estimated sensitivity of 75%, specificity of 68%, and accuracy of 72%; the area under the receiver operating characteristic curve was 0.75. In a community-based autopsy sample, this predictive model confirmed parkinsonism and hallucinations as important predictors of Lewy body pathology in patients with clinical Alzheimer's disease. The model also identified other demographic and clinical characteristics that might enhance the prediction of Lewy body pathology. PMID- 17085758 TI - Assessment of anxiety in dementia: an investigation into the association of different methods of measurement. AB - The objectives were to compare patient and caregiver ratings of anxiety for individuals with Alzheimer's disease and investigate their association with ratings of patient depression. Participants were 95 community-dwelling Alzheimer's disease patients and their caregivers. Measurements included caregiver proxy ratings: Rating Anxiety in Dementia, the NeuroPsychiatric Inventory, the Revised Memory and Behavior Problems Checklist, and the Cornell Scale for Depression in Dementia. Patient self-assessments included the Clinical Anxiety Scale (adapted) and the Geriatric Depression Scale. Correlations among anxiety measures ranged from rho = .16 to .40, whereas correlations between anxiety and depression scales ranged from rho = .03 to .66. Overlap among items across both anxiety and depression scales appeared to be the most important factor in determining these associations. Current scales for anxiety in dementia differ in their construction of symptoms and can overlap considerably with depression scales. Because of this, measures inadequately distinguish anxiety from depression in individuals with Alzheimer's disease. The measurement of anxiety in dementia needs additional refinement. PMID- 17085759 TI - Bereavement in cognitively impaired older adults: case series and clinical considerations. AB - The intersection of bereavement and cognitive impairment presents unique clinical challenges that have been overlooked in the literature. Cognitive impairment can interfere with normal ways of coping in the aftermath of loss. Elderly persons with cognitive impairment may have difficulty processing their loss and may even forget that their loved one has died, repeatedly asking other family members when the deceased is coming to visit. This can prove devastating for such individuals and their families, who are forced to keep reexperiencing their grief. This article examines the phenomenology of bereavement-related distress in older adults with cognitive impairment and dementia through a case series gathered from a large geriatric facility. The dilemmas in caring for bereaved elderly with cognitive difficulties and their families are highlighted, drawing attention to a vulnerable population. The purpose of this undertaking is to present a novel classification of a clinically relevant but overlooked issue as well as provide strategies for management and suggestions for future research. PMID- 17085760 TI - ACE clock scoring: a comparison with eight standard correction methods in a population of low educational level. AB - The authors compared the accuracy of Addenbrooke's Cognitive Examination Clock Drawing Test scoring method with 8 standard Clock Drawing Test scoring methods in a Spanish speaking population of low educational level. A clinical group composed of 70 patients affected by dementia and 25 patients with memory complaints without dementia was compared with 72 controls matched for gender, age, and educational level. The clinical group was studied with standard neuropsychological instruments and neuroimaging. Clock Drawing Tests extracted from Addenbrooke's Cognitive Examination were randomly distributed and scored by 2 independent raters. There were statistically significant differences between the different scoring methods. Addenbrooke's Cognitive Examination Clock Drawing Test scoring method obtained and optimal areas under the curve and interrater reliability, showing no statistical differences with previously standardized systems. In our population, Addenbrooke's Cognitive Examination and the methods of Cahn, Mendez, Rouleau, Shulman, and Sunderland were of optimal and equivalent usefulness regarding dementia detection. PMID- 17085761 TI - The Cambridge Behavioral Inventory: validation and application in a memory clinic. AB - The authors examined the validity of the Cambridge Behavioral Inventory (CBI), a questionnaire investigating broad neuropsychiatric symptoms and everyday functional ability in dementia. Test-retest reliability of the CBI was acceptable. Cross-validation with the Neuropsychiatric Inventory showed good concurrent validity of the CBI. The CBI reliably demonstrated that disinhibition, stereotypic behavior, elation, anxiety, poor self-care, and changes in eating habits occurred more commonly in patients with frontotemporal lobar degeneration than those with Alzheimer's disease. The authors concluded that the CBI is a reliable informant-based assessment of neuropsychiatric symptoms and everyday functioning and may be a suitable tool for use in general clinical practice settings. PMID- 17085762 TI - Factors associated with frequent admissions to an acute geriatric psychiatric inpatient unit. AB - As a first step toward developing strategies to reduce the frequency of psychiatric hospitalizations, the authors retrospectively collected and analyzed demographic and clinical variables from 424 consecutive admissions to a university-based geriatric psychiatry inpatient unit over a 20-month period. The study sample was dichotomized into patients who were admitted more than one time (35.6%) versus those with a single admission. Factors associated with rehospitalization were examined with multivariate logistic regression analysis. The great majority of readmissions (81%) occurred in the first 3 months after discharge. The logistic regression model indicated that significant predictors of rehospitalization were single relationship status, male gender, and bipolar disorder diagnosis. Our findings overlap with findings from previous similar studies and suggest that information readily obtainable on admission to an acute geriatric psychiatry inpatient unit may provide a useful indication of risk for frequent psychiatric hospitalizations and may contribute to readmission prevention strategies. PMID- 17085763 TI - Predictors of adherence within an intervention research study of the at-risk older drinker: PRISM-E. AB - The aim of this study was to determine predictors of research adherence and treatment initiation in at-risk older drinkers. This investigation was conducted at primary care clinics in the Philadelphia Veteran Affairs Medical Center and the University of Pennsylvania, participating sites in a larger multisite study trial (PRISM-E). Persons aged 65 and older with appointments at participating clinics were eligible for recruitment (n = 8367). Approximately half (n = 4000) consented to the study, of which 145 were identified as at-risk drinkers and 125 agreed to treatment. Slightly more than half of the patients who agreed to treatment attended a mental health visit. The results suggest that predictors of research adherence and treatment initiation vary across research stage. Principal predictors include age, mental health status, and at-risk drinking attributes. Moreover, there was evidence that an integrated care treatment model may be capable of improving treatment initiation in at-risk older drinkers who have no history of substance management behaviors. Future researchers can use the current findings to create mechanisms to improve research participation and treatment initiation and target participants with classifications of poor adherence. PMID- 17085765 TI - AtATG genes, homologs of yeast autophagy genes, are involved in constitutive autophagy in Arabidopsis root tip cells. AB - In Arabidopsis root tips cultured in medium containing sufficient nutrients and the membrane-permeable protease inhibitor E-64d, parts of the cytoplasm accumulated in the vacuoles of the cells from the meristematic zone to the elongation zone. Also in barley root tips treated with E-64, parts of the cytoplasm accumulated in autolysosomes and pre-existing central vacuoles. These results suggest that vacuolar and/or lysosomal autophagy occurs constitutively in these regions of cells. 3-Methyladenine, an inhibitor of autophagy, inhibited the accumulation of such inclusions in Arabidopsis root tip cells. Such inclusions were also not observed in root tips prepared from Arabidopsis T-DNA mutants in which AtATG2 or AtATG5, an Arabidopsis homolog of yeast ATG genes essential for autophagy, is disrupted. In contrast, an atatg9 mutant, in which another homolog of ATG is disrupted, accumulated a significant number of vacuolar inclusions in the presence of E-64d. These results suggest that both AtAtg2 and AtAtg5 proteins are essential for autophagy whereas AtAtg9 protein contributes to, but is not essential for, autophagy in Arabidopsis root tip cells. Autophagy that is sensitive to 3-methyladenine and dependent on Atg proteins constitutively occurs in the root tip cells of Arabidopsis. PMID- 17085766 TI - Evaluation of 3-deaza-adenosine analogues as ligands for adenosine kinase and inhibitors of Mycobacterium tuberculosis growth. AB - OBJECTIVES: Analyse a series of halogenated 3-deaza-adenosine analogues for efficacy against Mycobacterium tuberculosis H37Ra and determine if adenosine (Ado) kinase plays a role in the mechanism of action of these compounds. METHODS: The MIC as determined by microdilution broth assay provided a measure of antitubercular efficacy. MIC values were measured in M. tuberculosis strains H37Ra, SRICK1 (an Ado kinase-deficient strain of M. tuberculosis derived from H37Ra) and SRICK1 complemented with adoK, the gene which codes for Ado kinase in M. tuberculosis, in order to determine if Ado kinase played a role in the mechanism of action of these compounds. Furthermore, each compound was analysed as both a substrate and inhibitor for purified Ado kinases from M. tuberculosis and human sources. RESULTS: 2-Fluoro-3-deaza-adenosine, 3-fluoro-3-deaza adenosine and 2,3-difluoro-3-deaza-adenosine exhibited antitubercular activity that was Ado kinase-dependent. Furthermore, these compounds were at least 10-fold better substrates for M. tuberculosis Ado kinase than the human homologue. CONCLUSIONS: The Ado kinase-dependent antitubercular activity exhibited by several of the halogenated 3-deaza-adenosine analogues investigated in this study warrants further investigation of these compounds as antitubercular agents. Furthermore, substrate and inhibition studies provided insight into the Ado binding domain of Ado kinase, indicating that steric hindrance may limit the size of exocyclic modifications at the 3-position of Ado. PMID- 17085767 TI - Molecular analysis of constitutively expressed erm(C) genes selected in vitro in the presence of the non-inducers pirlimycin, spiramycin and tylosin. AB - OBJECTIVES: As known from other lincosamides and 16-membered macrolides, the antimicrobial agents pirlimycin, spiramycin and tylosin, which are frequently used for the therapy of bovine mastitis, cannot induce the expression of the resistance gene erm(C). The aim of this study was to confirm the ability of these three non-inducers to select for constitutively expressed erm(C) mutants in Staphylococcus aureus. METHODS: A S. aureus strain carrying an inducibly expressed erm(C) gene was incubated on agar plates containing inhibitory concentrations of each of the three antimicrobial agents. The erm(C) regulatory region of mutants obtained after overnight incubation was amplified by PCR; selected amplicons were sequenced and compared with the wild-type sequence. RESULTS: Mutants developed in the presence of each of the three antimicrobial agents. Constitutive expression of erm(C) was due to variations in the erm(C) regulatory region. A total of 10 different types of deletions ranging in size between 16 and 121 bp as well as 20 different types of duplications ranging between 24 and 602 bp were detected. The frequencies by which sequence alterations occurred as well as the types of alterations detected varied with regard to the antimicrobial agents used for selection. CONCLUSIONS: All sequence alterations observed explained constitutive erm(C) gene expression by functional inactivation of translational attenuation. In order to prevent the development of constitutively resistant isolates under therapy, the results of this study support the recommendation not to use lincosamides or 16-membered macrolides for the control of staphylococcal infections caused by strains harbouring inducibly expressed erm(C) genes. PMID- 17085768 TI - Antimicrobial effectiveness of ketoconazole against metronidazole-resistant Helicobacter pylori isolates from Iranian dyspeptic patients. PMID- 17085769 TI - Identification of two novel quantitative trait loci for pre-eclampsia susceptibility on chromosomes 5q and 13q using a variance components-based linkage approach. AB - Pre-eclampsia/eclampsia (PE/E) is a common and serious disorder of human pregnancy that is associated with substantial maternal and perinatal morbidity and mortality. The suspected aetiology of PE/E is complex, with susceptibility being attributable to multiple environmental factors and a large genetic component. By assuming that the underlying liability towards PE/E susceptibility is inherently quantitative, any PE/E susceptibility gene would represent a quantitative trait locus (QTL). This assumption enables a more refined and powerful variance components procedure using a threshold model for our PE/E statistical analysis. Using this more efficient linkage approach, we have now re analysed our previously completed Australian/New Zealand genome scan data to identify two novel PE/E susceptibility QTLs on chromosomes 5q and 13q. We have obtained strong evidence of linkage on 5q with a peak logarithm-of-odds (LOD) score of 3.12 between D5S644 and D5S433 [at approximately 121 centimorgan (cM)] and strong evidence of linkage on 13q with a peak LOD score of 3.10 between D13S1265 and D13S173 (at approximately 123 cM). Objective identification and prioritization of positional candidate genes using the quantitative bioinformatics program GeneSniffer revealed highly plausible PE/E candidate genes encoding aminopeptidase enzymes and a placental peptide hormone on the 5q QTL and two type IV collagens on the 13q QTL regions, respectively. PMID- 17085770 TI - Genomic structure and tissue-specific expression of human and mouse genes encoding homologues of the major bovine seminal plasma proteins. AB - Sperm capacitation is a maturation event that takes place in the female reproductive tract and is essential for fertilization. A family of phospholipid binding proteins present in bovine seminal plasma (BSP proteins) binds the sperm membrane at ejaculation and promotes bovine sperm capacitation. Homologues of these proteins have also been isolated from boar, ram, goat, bison and stallion seminal fluid, suggesting that BSP proteins and their homologues are conserved among mammals. However, there have been no reports on BSP-homologous proteins in mice and humans to date. A search of the mouse and human genomes, using the nucleic acid sequences of BSP proteins, revealed the presence of three BSP-like sequences in the mouse genome, named mouse BSP Homologue 1 (mBSPH1), mBSPH2 and mBSPH3, and one sequence in the human genome (hBSPH1). Mouse epididymal expressed sequence tags corresponding to partial sequences of mBSPH1 and mBSPH2 were identified. The entire complementary DNA (cDNA) sequences of mBSPH1 and mBSPH2 from mouse epididymis and hBSPH1 from human epididymis were obtained by 5'-/3' rapid amplification of cDNA ends (RACE) and encode predicted proteins containing two tandemly repeated fibronectin type II domains, which is the signature of the BSP family of proteins. Using RT-PCR, it was revealed that mBSPH1, mBSPH2 and hBSPH1 mRNA are expressed only in the epididymis. Expression of mBSPH3 was not detected in any tissue and probably represents a pseudogene. This work shows, for the first time, that BSP homologues are expressed in mouse and human and may be involved in sperm capacitation in these species. PMID- 17085771 TI - Immunophenotyping of chimeric cells in localized scleroderma. AB - OBJECTIVE: Localized scleroderma causes thickening of the skin due to excessive collagen deposition. This condition has clinical and histopathological similarities to chronic graft-vs-host disease. We wanted to identify whether chimeric cells are present in the affected tissue in localized scleroderma and to further investigate the role of chimerism by immunophenotyping the chimeric cells. We hypothesize that the presence of chimerism and immunotypic chimeric cells will lend to an understanding of the pathogenesis of localized scleroderma and possible mechanisms by which chimeric cells participate in autoimmunity. METHODS: We studied skin biopsies from 18 localized scleroderma patients and compared them with concurrent biopsies from unaffected skin in a subset of patients. Skin biopsies from morphoea and linear scleroderma patients were analysed for the presence of chimeric cells using male-female (X, Y) differences. Cell surface markers (CD4, CD8, CD19/20, CD68, S100, CD14 and CD56) were determined for cell phenotyping of chimeric cells. RESULTS: Overall, the affected tissue contained a greater number of lymphocytic inflammatory cells. In the affected tissue, 38% of the total chimeric cells were CD68+ (dendritic cell, monocyte and macrophage marker), 29% Langerin/S100+ (dendritic cell marker), 26% CD8+ (cytotoxic T-lymphocyte marker), 20% CD19/20+ (B-lymphocyte marker), 14% CD4+ (T-helper lymphocyte) and 0% CD56+ (natural killer cell marker). CONCLUSIONS: We report that not only are chimeric cells present in affected localized scleroderma lesions but they also are more likely to be dendritic cells and B lymphocytes suggesting a role in the pathogenesis of localized scleroderma. PMID- 17085772 TI - The role of psychosocial factors in predicting the onset of chronic widespread pain: results from a prospective population-based study. AB - OBJECTIVE: Chronic widespread pain (CWP) is strongly associated with psychosocial distress both in a clinical setting and in the community. The aim of this study was to determine the contribution of measures of psychosocial distress, health seeking behaviour, sleep problems and traumatic life events to the development of new cases of CWP in the community. METHODS: In a population-based prospective study, 3171 adults aged 25-65 yrs free of CWP were followed-up 15 months later to identify those with new CWP. Baseline data were available on their scores from a number of psychological scales including Illness Attitude Scales (IAS), Somatic Symptom Checklist (SSC), Hospital Anxiety & Depression Scale, Sleep Problems Scale, and Life Events Inventory. RESULTS: 324 subjects [10%, 95% confidence interval (CI) 9.2, 11.3] developed new CWP at follow-up. After adjustment for age and sex, three factors independently predicted the development of CWP: scoring three or more on the SSC [odds ratio (OR) 1.8, 95% CI 1.1, 3.1], scoring eight or more on the Illness Behaviour subscale of the IAS (OR 3.3, 95% CI 2.3, 4.8), and nine or more on the Sleep Problem Scale (OR 2.7, 95% CI 1.6, 3.2). Subjects exposed to all three factors were at 12 times the odds of new CWP than those with low scores on all scales. CONCLUSION: Subjects are at substantial increased odds of developing CWP if they display features of somatization, health-seeking behaviour and poor sleep. Psychosocial distress has a strong aetiological influence on CWP. PMID- 17085773 TI - Safety and efficacy of black cohosh (Cimicifuga racemosa) during pregnancy and lactation. AB - BACKGROUND: There is a lack of basic knowledge on the part of both clinicians and patients as to the indications for use and safety of herbal medicines in pregnancy and lactation. This is one article in a series that systematically reviews the evidence for commonly used herbs during pregnancy and lactation. OBJECTIVES: To systematically review the literature for evidence on the use, safety, and pharmacology of black cohosh focusing on issues pertaining to pregnancy and lactation. METHODS: We searched 7 electronic databases and compiled data according to the grade of evidence found. RESULTS: Black cohosh, alone or in combination with other medicinal herbs as "mother's cordial", has a long traditional use and is frequently used by midwives as a uterine stimulant and labour-inducing aid. Low-level evidence based on theory and expert opinion shows the following concerns with respect to black cohosh use during pregnancy: 1) labour-inducing effects, 2) hormonal effects, 3) emmenagogue properties and, 4) anovulatory effects. During lactation, there is low-level evidence from theoretical and expert opinion of black cohosh having hormonal properties. CONCLUSIONS: Black cohosh should be used with caution during pregnancy, particularly during the first trimester where its purported labour-inducing effects could be of concern, and during lactation. Black cohosh should undergo rigorous high quality human studies to determine its safety in pregnancy and lactation. PMID- 17085774 TI - Safety and efficacy of echinacea (Echinacea angustafolia, e. purpurea and e. pallida) during pregnancy and lactation. AB - BACKGROUND: There is a lack of basic knowledge on the part of both clinicians and patients as to the indications for use and safety of herbal medicines used in pregnancy and lactation. This is one article in a series that systematically reviews the evidence for commonly used herbs during pregnancy and lactation. OBJECTIVES: To systematically review the literature for evidence on the use, safety, and pharmacology of echinacea focusing on issues pertaining to pregnancy and lactation. METHODS: We searched 7 electronic databases and compiled data according to the grade of evidence found. RESULTS: There is good scientific evidence from a prospective cohort study that oral consumption of echinacea during the first trimester does not increase the risk for major malformations. Low-level evidence based on expert opinion shows that oral consumption of echinacea in recommended doses is safe for use during pregnancy and lactation. CONCLUSIONS: Echinacea is non-teratogenic when used during pregnancy. Caution with using Echinacea during lactation until further high quality human studies can determine its safety. PMID- 17085775 TI - Safety and efficacy of St. John's wort (hypericum) during pregnancy and lactation. AB - BACKGROUND: There is a lack of basic knowledge on the part of both clinicians and patients as to the indications for use and safety of herbal medicines in pregnancy and lactation. This is one article in a series that systematically reviews the evidence for commonly used herbs during pregnancy and lactation. OBJECTIVES: To systematically review the literature for evidence on the use, safety, and pharmacology of St. John's wort focusing on issues pertaining to pregnancy and lactation. METHODS: We searched 7 electronic databases and compiled data according to the grade of evidence found. RESULTS: There is very weak scientific evidence based on a case report that St Johns wort is of minimal risk when taken during pregnancy. There is in vitro evidence from animal studies that St John's wort during pregnancy does not affect cognitive development nor cause long-term behavioral defects, but may lower offspring birth weight. There is weak scientific evidence that St. John's wort use during lactation does not affect maternal milk production nor affect infant weight, but, in a few cases, may cause colic, drowsiness or lethargy. There is weak scientific evidence that St John's wort induces CYP450 enzymes, which may lower serum medication levels below therapeutic range; this may be of concern when administering medications during pregnancy and lactation. CONCLUSIONS: Caution is warranted with the use of St John's wort during pregnancy until further high quality human research is conducted to determine its safety. St John's wort use during lactation appears to be of minimal risk, but may cause side effects. Caution is warranted when using medications along with St John's wort. PMID- 17085776 TI - Safety and efficacy of ginkgo (Ginkgo biloba) during pregnancy and lactation. AB - BACKGROUND: There is a lack of basic knowledge on the part of both clinicians and patients as to the indications for use and safety of herbal medicines used in pregnancy and lactation. This is one article in a series that systematically reviews the evidence for commonly used herbs during pregnancy and lactation. OBJECTIVES: To systematically review the literature for evidence on the use, safety, and pharmacology of ginkgo focusing on issues pertaining to pregnancy and lactation. METHODS: We searched 7 electronic databases and compiled data according to the grade of evidence found. RESULTS: There is some very weak scientific evidence from animal and in vitro studies that ginkgo leaf has antiplatelet activity, which may be of concern during labour as ginkgo use could prolong bleeding time. Low-level evidence based on expert opinion shows that ginkgo leaf may be an emmenagogue and have hormonal properties. The safety of ginkgo leaf during lactation is unknown. Patients and clinicians should be aware of past reports of ginkgo products being adulterated with colchicine. CONCLUSIONS: Ginkgo should be used with caution during pregnancy, particularly around labour where its anti-platelet properties could prolong bleeding time. During lactation the safety of ginkgo leaf is unknown and should be avoided until high quality human studies are conducted to prove its safety. PMID- 17085777 TI - The effect of a short tutorial on the incidence of prescribing errors in pediatric emergency care. AB - BACKGROUND: In the paediatric emergency department (ED) trainees are more likely to commit prescribing errors. OBJECTIVE: To determine whether a short educational intervention reduces the incidence of prescribing errors among trainees in a pediatric ED. METHODS: A prospective cohort study at the ED of a tertiary paediatric hospital. All fellows and residents arriving at the ED at the beginning of the academic year were invited to participate in a 30-minute tutorial focusing on appropriate methods for prescribing medications, followed by a written test. Eighteen days were selected randomly during July 2001. All the charts from these days were reviewed for medication errors. Two reviewers, blinded to whether or not a particular physician attended the tutorial, independently decided whether or not an error had occurred. The main outcome measure was the number of prescribing errors. RESULTS: Twenty-two trainees worked in the ED during July 2001. Of these, 13 trainees attended the tutorial. Eight hundred and ninety nine orders given by trainees were evaluated. We identified 66 (12.4%) errors in 533 orders given by those who attended tutorial, and 46 (12.7%) errors in 363 orders given by those who did not attend tutorial. The adjusted odds of a medication error was not significantly different between those who did not attend the tutorial and those who did (OR: 1.07 95% CI: 0.66-1.70). CONCLUSIONS: A short tutorial, followed by a written test, administered to trainees before entering their rotation in the paediatric ED, did not appear to reduce prescribing errors. PMID- 17085778 TI - The innate immune facet of brain: human neurons express TLR-3 and sense viral dsRNA. AB - Inflammation is an important factor in the pathogenesis of neurodegenerative diseases, such as Alzheimer's disease or multiple sclerosis, and during microbial infections of the nervous system. Glial cells were thought to be the main contributor for cytokine and chemokine production and Toll-like receptor (TLR) expression in the brain. Here, we report that human neurons express TLR-3, a major receptor in virus-mediated innate immune response. We established that these cells can mount a strong inflammatory response characterized by the expression of inflammatory cytokines (TNF-alpha, IL-6), chemokines (CCL-5 and CXCL-10), and antiviral molecules (2'5'OAS and IFN-beta) after treatment with dsRNA - a by-product of viral infection and ligand of TLR-3. This work firmly establishes that human neurons, in absence of glia, have the intrinsic machinery to trigger robust inflammatory, chemoattractive, and antiviral responses. PMID- 17085779 TI - Prions: protein only or something more? Overview of potential prion cofactors. AB - Transmissible spongiform encephalopathies (TSEs) in humans and animals are attributed to protein-only infectious agents, called prions. Prions have been proposed to arise from the conformational conversion of the cellular protein PrP(C) into a misfolded form (e.g., PrP(Sc) for scrapie), which precipitates into aggregates and fibrils. It has been proposed that the conversion process is triggered by the interaction of the infectious form (PrP(Sc)) with the cellular form (PrP(C)) or might result from a mutation in the gene for PrP(C). However, until recently, all efforts to reproduce this process in vitro had failed, suggesting that host factors are necessary for prion replication. In this review we discuss recent findings such as the cellular factors that might be involved in the conformational conversion of prion proteins and the potential mechanisms by which they could operate. PMID- 17085780 TI - Role of DJ-1 in Parkinson's disease. AB - Parkinson's disease (PD), one of the most common neurodegenerative diseases, is a multifactorial disease caused by both genetic and environmental factors. Although most patients suffering from PD have a sporadic disease, several genetic causes have been identified in recent years, including alpha-synuclein, parkin, PINK1, dardarin (LRRK2), and DJ-1. DJ-1 deletions and point mutations have been found worldwide, and loss of functional protein was shown to cause autosomal recessive PD. Moreover, DJ-1 immunoreactive inclusions are found in other alpha synucleopathies and tauopathies, indicating that different neurodegenerative diseases might share a common mechanism in which DJ-1 might play a key role. The function of DJ-1 is still unknown; however, it is associated with various cellular processes, including response to oxidative stress, cellular transformation, RNAbinding, androgen-receptor signaling, spermatogenesis, and fertilization. This article reviews the current knowledge on DJ-1, focusing on its importance in the pathogenesis of PD. PMID- 17085781 TI - CoREST-like complexes regulate chromatin modification and neuronal gene expression. AB - The mammalian CoREST ([co]repressor for element-1-silencing transcription factor) complex was first identified associated with the repressor for element-1 silencing transcription factor (REST)/neuronal restrictive silencing factor. The CoREST complex is a chromatin-modifying corepressor complex that acts with REST to regulate neuronal gene expression and neuronal stem cell fate. Components of a CoREST-like complex have been identified recently in Xenopus laevis, Caenorhabditis elegans, and Drosophila melanogaster. Like the mammalian complex, the Drosophila complex is required to regulate neuronal gene expression, whereas the C. elegans homologs regulate the expression of the hop-1 presenilin gene, suggesting an ancient conserved function of CoREST complexes in regulating neuronal gene expression. PMID- 17085782 TI - Receptor protein tyrosine phosphatase from stem cells to mature glial cells of the central nervous system. AB - The cornerstone of cell signaling is largely based on the phosphorylation state that is defined by the equilibrium of the activity of protein kinases and protein phosphatases. The role of protein tyrosine kinases in brain development, brain tumors, and neurodegenerative diseases was studied extensively, yet, the importance of protein tyrosine phosphatases (PTPs) in the development of glial cells was somewhat neglected. In this review, we have summarized recent findings of PTP expression during development of the central nervous system and the different cell types of the brain, from stem cells to mature glial cells, and highlighted the potential role of these enzymes in neuronal stem cell development, glioblastomas, and myelination. PMID- 17085783 TI - ErbB4 activation inhibits MPP+-induced cell death in PC12-ErbB4 cells: involvement of PI3K and Erk signaling. AB - The neuroprotective effects of neuregulin (NRG), a polypeptide growth factor, on 1-methyl-4-phenylpyridinium ion (MPP+)-induced cell death and oxidative stress in PC12-ErbB4 cells were investigated. Treatment of PC12-ErbB4 cells with MPP+ induced cell death that was markedly attenuated by NRG. The PI3K/PKB/Akt and Ras/MapK signaling pathways probably mediate the survival effect of NRG. NRG induces prolonged activation of PKB/Akt and Erk. Moreover, inhibition of the PI3K and MEK activities prevented the NRG-induced survival effect. Overexpression of constitutively active PI3K or H-Ras (12V) inhibited MPP+-mediated cell death. In addition, MPP+- mediated reactive oxygen species (ROS) elevation was also inhibited by NRG. The effect of NRG on ROS levels was blocked by PI3K and MEK inhibitors, indicating that both signaling pathways can regulate the toxic ROS levels induced by MPP+. Taken together, these results indicate that in PC12-ErbB4 cells, the NRG-induced neuroprotective effect from MPP+ treatment, requires PI3K/PKB/Akt and Ras/MapK signaling networks. PMID- 17085784 TI - Levels of mRNA coding for alpha-, beta-, and gamma-synuclein in the brains of newborn, juvenile, and adult rats. AB - Synucleins are proteins known for their malfunction in a group of illnesses called synucleopathies, which includes Alzheimer's and Parkinson's disease. To learn more about the role of synucleins in the CNS, we have studied levels of message coding for alpha-, beta-, and gamma-synuclein using quantitative RT-PCR. Levels of synuclein mRNAs were studied in the cerebral cortex (left and right, anterior and posterior), hippocampus, striatum, and cerebellum, obtained from 5-d old (newborn), 1-mo (juvenile)-, and 6-, and 9-mo (adult)-old rats. The mRNA levels for all synucleins varied significantly among structures. The rank order of mRNA levels in different structures was cortex = hippocampus > striatum > cerebellum for alpha-synuclein; cortex > hippocampus = cerebellum > striatum for beta-synuclein; and hippocampus = striatum > cortex = cerebellum for gamma synuclein. There was significant effect of age for mRNA levels for all synucleins. The dynamics of these changes were different depending on type of synuclein and brain structure. Levels of mRNA for alpha-synuclein were significantly reduced with age in all structures except hippocampus. For beta- and gamma-synuclein, levels increased significantly only in the cerebral cortex and only from 5 d to 1 mo of age. In contrast, gamma-synuclein levels in the cerebellum were very high at 5 d and significantly reduced at 1 mo of age. The revealed pattern and dynamics of changes in the levels of mRNA coding for synucleins would support the conclusion for an important role of these molecules during development and the aging process. PMID- 17085785 TI - NF-kappaB-associated MnSOD induction protects against beta-amyloid-induced neuronal apoptosis. AB - Expression of manganese superoxide dismutase (MnSOD), a nuclear-encoded mitochondrial primary antioxidant enzyme, is protective against various paradigms of oxidative stress-induced brain injury. We have shown previously that the presence of an intronic nuclear factor site, kappaB (NF-kappaB), in the MnSOD gene is essential for the induction of MnSOD by tumor necrosis factor alpha (TNF alpha). However, whether activation of NF-kappaB is protective against oxidative stress-induced neuronal injury is unclear. In the present study, we demonstrate that TNF-alpha activates NF-kappaB activity in neuronal, SH-SY5Y, cells and preferentially enhances the binding of p50 and p65 to the promoter/enhancer regions of the MnSOD gene. Binding of NF-kappaB members to the MnSOD gene leads to the induction of MnSOD mRNA and protein levels. Consequently, induction of MnSOD by TNF-alpha primes neuronal cells to develop resistance against subsequent exposure to beta-amyloid and FeSO(4). Taken together, these results suggest that NF-kappaB might exert its protective function by induction of MnSOD leading to subsequent protection against oxidative stress-induced neuronal injury. PMID- 17085787 TI - Current issues with beta2-adrenoceptor agonists: historical background. AB - The discovery that dessicated adrenal glands had beneficial effects in asthma arose in 1900 following a vogue for studying organotherapy at the end of the 19th century. The adrenal hormone adrenaline was found to have sympathomimetic properties and was isolated and synthesized in 1901. The first nonselective beta agonist, isoproterenol, was isolated in 1940, followed by the development of selective beta2-agonists in the 1960s and the introduction of the long-acting beta2-agonists in the 1990s. The introduction of beta2-selectivity reduced adverse effects, as did developments in inhaler technology that allowed subjects to inhale much smaller doses of drug selectively to the airways. The beta2 agonists are some of the more important drugs to have been developed in the 20th century. Excessive doses can cause problems, and attempts to maximize the benefit from beta2-agonists and to reduce adverse effects has led to considerable epidemiological, clinical, and mechanistic research over the last 50 yr. PMID- 17085788 TI - Current issues with beta2-adrenoceptor agonists: pharmacology and molecular and cellular mechanisms. AB - Beta2-adrenoceptors are widely, almost ubiquitously, expressed. Activation of these receptors on bronchial smooth muscle by short- and long-acting beta2 adrenoceptor agonists causes bronchodilation. Here, the beta2-adrenoceptor is linked by the G protein, Gs, to adenylyl cyclase, which increases cyclic adenosine monophosphate (cAMP), thus activating protein kinase A, which affects calcium levels and reduces the efficiency of myosin light-chain kinase, causing relaxation. Activation also entrains numerous acute and longer term downregulation responses affecting the number, location, and net efficiency of signaling of the receptor. Synthetic beta2-agonists are all "partial agonists," incompletely able to optimally stimulate cAMP signal transduction. However, compared with some cells (such as mast cells) involved in exercise- induced asthma induction, airway smooth muscle is privileged in that transduction efficiency is intrinsically high and the tissue is very resistant to complete downregulation. Glucocorticosteroids have broadly beneficial interactions with beta2-adrenoceptors. Researchers have recently discovered that the beta2 adrenoceptor may function as a homodimer and that it can form heterodimers with both the beta1- and beta3-adrenoceptors, and possibly other receptors. This further complicates interpretation of the effect of beta2-adrenoceptor polymorphisms, but it is unknown whether this occurs in humans in vivo. Researchers have known for some time that strong contraction involving receptors coupled to the Gq G protein (e.g., cholinergic and leukotriene receptors via negative biochemical crosstalk), virus infection (via uncoupling), and inflammation (via kinases) can impair relaxation. Most recently, researchers have discovered that the beta2-adrenoceptor can also send potentially adverse signals after "atypical coupling" to Gq rather than Gs. The clinical implications of these uncouplings, crosstalk, and atypical coupling possibilities are not well understood. PMID- 17085789 TI - Regulation of mast cells by beta-agonists. AB - The human lung mast cell is known to be a critical effector cell in the mediation of asthma. Activation of the mast cell by allergens and other stimuli leads to the release and generation of a wide variety of autacoids that cause bronchoconstriction, promote inflammation, and may influence airway remodeling. Therefore, the stabilization of mast cells has obvious value in the prevention of asthma. Among the drugs used to treat asthma, only beta-agonists are effective stabilizers of mast cells. Both short- and long-acting beta-agonists are effective against mast cells, but there are differences between agonists regarding the extent of inhibitory activity attained. Consequently, the type of beta-agonist prescribed influences the degree of mast cell stabilization possible. Despite the potential value of attenuating mast cell activity with beta agonists, this benefit may diminish with time because of the development of tolerance. Both short- and long-acting beta-agonists can induce tolerance to mast cell stabilization, and generally, higher efficacy agonists tend to induce greater levels of tolerance; however, weaker agonists induce greater levels of tolerance than might be expected. Tolerance to the mast-cell-stabilizing effects of beta- agonists may be an issue clinically, because this occurs more readily than tolerance to smooth muscle relaxation. This could lead to a situation in which beta-agonists fail to prevent the release of mediators from mast cells but can still effectively relax airway smooth muscle. The continued ability to bronchodilate could mask the unfavorable consequences of unchecked mediator release from mast cells. PMID- 17085790 TI - Beta2-agonists and bronchial hyperresponsiveness. AB - Bronchial hyperresponsiveness (BHR) is a characteristic feature of asthma, and individuals with this disease respond to a range of physiological and chemical insults that are otherwise innocuous to healthy subjects, suggesting that the mechanisms underlying this phenomenon are characteristic of the asthma phenotype. BHR can be increased following exposure to environmental allergens in suitably sensitized individuals, pollutants, and certain viruses and can also be exacerbated by exposure to certain drugs, including nonsteroidal anti inflammatory agents and beta-blockers. Although beta2-agonists administered acutely remain the treatment for the symptoms of asthma, paradoxically, regular treatment with these drugs can result in an increase in BHR, and this has been suggested to contribute to the increase in asthma morbidity and mortality that has been reported by numerous investigators. This article highlights our current understanding of this phenomenon and examines the potential mechanisms responsible for this effect. PMID- 17085791 TI - Beta2-agonists and exercise-induced asthma. AB - Beta2-agonists taken immediately before exercise provide significant protection against exercise- induced asthma (EIA) in most patients. However, when they are taken daily, there are some negative aspects regarding severity, control, and recovery from EIA. First, there is a significant minority (15-20%) of asthmatics whose EIA is not prevented by beta2-agonists, even when inhaled corticosteroids are used concomitantly. Second, with daily use, there is a decline in duration of the protective effect of long-acting beta2-agonists. Third, if breakthrough EIA occurs, recovery of lung function is slower in response to a beta2-agonist, and additional doses are often required to achieve pre-exercise values. If a person who takes a beta2-agonist daily experiences problems with exercise, then the physician should consider changing the treatment regimen to achieve better control of EIA. These problems likely result from desensitization of the beta2 receptor on the mast cell, which enhances mediator release, and on the bronchial smooth muscle, which enhances the bronchoconstrictor response and delays recovery from EIA. These effects are reversed within 72 h after cessation of a beta2 agonists. The important clinical question is: Are we actually compromising the beneficial effects of beta2-agonists on the prevention and recovery from EIA by prescribing them daily? Patients with EIA need to ensure that their doses of inhaled corticosteroid or other anti-inflammatory therapy are optimized so that, if necessary, a beta2-agonist can be used intermittently as prophylactic medication with greater confidence in the outcome. PMID- 17085792 TI - Recovery from bronchoconstriction and bronchodilator tolerance. AB - Until recently, researchers believed that tolerance or tachyphylaxis to the bronchodilator effects of beta-agonists did not occur. However, recent studies examining the recovery from bronchoconstriction have clearly shown that an impaired response to beta-agonists occurs in patients who have been using regular beta-agonist treatment. This tolerance develops with both long- and short-acting beta-agonists and is not affected by treatment with inhaled steroids. It develops rapidly, reaching a maximum within 1 wk of starting beta-agonists, and has been demonstrated after methacholine, hypertonic saline, mannitol, and exercise induced bronchoconstriction. The observed reduction in the bronchodilator response is proportional to the severity of bronchoconstriction. Therefore, although individuals with stable asthma show little evidence of tolerance, those with severe bronchospasm have a markedly reduced bronchodilator response to beta agonists. Almost all asthmatics show evidence of tolerance when tested in the setting of bronchoconstriction, although the extent of this tolerance varies. The reasons for this interindividual variation are not understood. Bronchodilator tolerance is difficult to study in the clinical setting because nearly every patient has used multiple doses of beta-agonist before seeking medical attention. However, there is compelling evidence that the response to rescue beta-agonist treatment is reduced in those who use regular long- or short-acting beta agonists. The extent to which this phenomenon contributes to asthma morbidity and mortality remains to be determined. PMID- 17085793 TI - Clinical concerns with inhaled beta2-agonists: adult asthma. AB - Inhaled beta2-agonists, when used regularly, cause subtle but significant worsening of asthma control. Overuse of inhaled beta2-agonists is associated with increased risk of death from asthma in a dose-response fashion. beta2-Agonists enhance airway responses to allergens, including induced airway hyperresponsiveness and induced airway inflammation. This is a plausible explanation for beta2-agonist-worsened asthma control. These direct effects of inhaled beta2-agonists, including increased airway response to allergen, tolerance, etc., may partially explain the association of overuse with asthma death. However, it is probable that the major reason for the association of beta2 agonists overuse and asthma mortality is an indirect effect. Inhaled beta2 agonists are effective relievers and preventers of bronchoconstriction and asthma symptoms but fail to treat the underlying pathogenesis, namely the airway inflammation. Thus, overuse may mask the true asthma severity and result in both an underappreciation and undertreatment of the disease. This would provide a rational explanation for the relationship of inhaled beta2-agonist use and mortality and also would fit the dose-response pattern. Inhaled beta2-agonists should be used exclusively as needed for relief of symptoms and their requirement should be infrequent: the need for excessive doses of beta2-agonists provides a useful marker of asthma (lack of) control. PMID- 17085794 TI - Beta2-agonists for asthma: the pediatric perspective. AB - Inhaled beta-agonists are commonly prescribed for the treatment of wheezing disorders in infants and children. Despite this, there are concerns that these medications have potentially detrimental effects on lung health and symptoms. We will review the ontogeny of beta-agonist receptor and smooth muscle development from fetal life through infancy and childhood as well as the evidence supporting the clinical utility of beta-agonists in wheezing infants and asthmatic children. Finally, the potential detrimental effects of long- and short-acting beta agonists in infants and children are discussed. PMID- 17085795 TI - Systematic review of clinical outcomes in chronic obstructive pulmonary disease: beta-agonist use compared with anticholinergics and inhaled corticosteroids. AB - Much controversy surrounds the use of beta-agonists in obstructive lung disease. Regular beta2- agonist use in asthma results in tolerance to its effects and an increase in asthma-related deaths. Less is known about clinical outcomes in chronic obstructive pulmonary disease (COPD). This systematic review and meta analysis evaluates the long-term effect of beta2-agonist use on severe exacerbations requiring hospitalization or trial withdrawal, respiratory deaths, and total mortality in patients with COPD. Results for beta2-agonists are compared with results for anticholinergics and inhaled corticosteroids. Pooled results from randomized controlled trials show that anticholinergics, such as tiotropium and ipratropium, significantly reduce severe exacerbations and respiratory deaths compared with placebo. Conversely, beta2-agonists increase respiratory deaths, probably because of tolerance that develops to their bronchodilator and bronchoprotective effects. Anticholinergics significantly reduce exacerbations and total mortality compared with beta-agonists. The combination of the two bronchodilators is not more effective than anticholinergics alone in improving long-term clinical outcomes. Inhaled corticosteroids significantly reduce severe exacerbations and the decline in lung function over time, without affecting mortality. In conclusion, inhaled anticholinergic bronchodilators and corticosteroids should be used to improve long-term clinical outcomes in patients with COPD. beta-Agonists increase respiratory deaths in COPD, possibly as a result of poorer disease control. PMID- 17085796 TI - Interactions between corticosteroids and beta2-agonists. AB - In vitro studies have demonstrated numerous ways in which beta2-agonists and corticosteroids may interact. Together with evidence of improved control of airway diseases using a combination therapy of inhaled corticosteroids and long acting beta-agonists compared with treatment with either drug alone, this suggests that there may be a beneficial synergy between these two classes of medication. However, a positive interaction has not been clearly demonstrated in vivo. There is little evidence that beta2-agonists enhance the anti-inflammatory actions of inhaled corticosteroids. Corticosteroids do not enhance the bronchodilator response to beta2- agonists, nor do they appear to prevent the development of tolerance during chronic beta2-agonist treatment. The evidence that high doses of corticosteroids can reverse tolerance to beta2-agonists is conflicting. Most of the clinical benefits from combinations using inhaled corticosteroids and long-acting beta-agonists to treat asthma could result from additive or complimentary effects of the drugs on different parts of the disease process. Nevertheless, there are several studies in which the combination of a long- or short-acting beta-agonist with an inhaled corticosteroid appears to have provided more than additive clinical benefits. The issue warrants further study. Combined inhaled corticosteroid/beta-agonist treatment will remain the basis of asthma management for the foreseeable future, and although the overall clinical benefit of the combination is no longer in doubt, there is more to learn about maximizing this benefit and minimizing adverse effects. PMID- 17085797 TI - Pharmacogenetics of beta2-agonist drugs in asthma. AB - Genetic variation in the structure of the beta2-adrenoceptor (ADRB2) is a potentially important source of variability in the response to beta2-agonist drugs. The relevant polymorphisms are common, with allele frequencies as high as 50% in some cases. "Nonsynonymous" variants result in amino acid substitutions in the receptor structure, of which substitution of Gly for Arg at position 16 and Glu for Gln at position 27 are the most clinically relevant. Other polymorphisms occur in the noncoding flanking regions of the gene and may be important in regulating ADRB2 expression, thus affecting individual drug responses. Linkage disequilibrium involving polymorphisms in both the coding and noncoding regions may explain the diversity of outcomes regarding functional in vitro studies. To date, analysis by haplotype, rather than genotype, has tended to confuse the issues. Functional in vitro studies provide the basis by which these polymorphisms are believed to be relevant in vivo. However, follow-up investigations have not substantiated the conventional wisdom that Gly-16 confers increased propensity to downregulation, whereas Glu-27 confers resistance. The in vivo picture is further complicated by the dynamic status of ADRB2s. Paradoxically, downregulation of receptors with exogenous beta2-agonist drugs may occur more readily in cells that are relatively resistant to downregulation with exposure to endogenous catecholamine. Numerous studies have highlighted that the Arg-16 variant is associated with an enhanced bronchodilator response in vivo, although the picture is not a consistent one. Outcomes during chronic treatment of asthma with beta2-agonists have been more consistent. In patients who are homozygous for Arg-16, reduced lung function and increased frequency of exacerbations occur during chronic treatment with short-acting beta2-agonists. With long-acting beta2-agonists, the overall clinical benefits may be worse for Arg-16 than for the Gly-16 genotype. Therefore, ADRB2 polymorphisms are an important consideration in assessing individual patients with difficult asthma that may be drug-induced for reasons of ADRB2 genotype. They are also relevant in the design of clinical trials involving beta2-agonist therapy. PMID- 17085798 TI - beta2-Agonists at the Olympic Games. AB - The different approaches that the International Olympic Committee (IOC) had adopted to beta2-agonists and the implications for athletes are reviewed by a former Olympic team physician who later became a member of the Medical Commission of the IOC (IOC-MC). Steadily increasing knowledge of the effects of inhaled beta2-agonists on health, is concerned with the fact that oral beta2-agonists may be anabolic, and rapid increased use of inhaled beta2-agonists by elite athletes has contributed to the changes to the IOC rules. Since 2001, the necessity for athletes to meet IOC criteria (i.e., that they have asthma and/or exercise induced asthma [EIA]) has resulted in improved management of athletes. The prevalence of beta2-agonist use by athletes mirrors the known prevalence of asthma symptoms in each country, although athletes in endurance events have the highest prevalence. The age-of-onset of asthma/EIA in elite winter athletes may be atypical. Of the 193 athletes at the 2006 Winter Olympics who met th IOC's criteria, only 32.1% had childhood asthma and 48.7% of athletes reported onset at age 20 yr or older. These findings lead to speculation that years of intense endurance training may be a causative factor in bronchial hyperreactivity. The distinction between oral (prohibited in sports) and inhaled salbutamol is possible, but athletes must be warned that excessive use of inhaled salbutamol can lead to urinary concentrations similar to those observed after oral administration. This article provides justification that athletes should provide evidence of asthma or EIA before being permitted to use inhaled beta2-agonists. PMID- 17085799 TI - Serious adverse events and death associated with treatment using long-acting beta agonists. AB - Two very large, randomized, double-blind clinical trials performed in the United Kingdom and in the United States have suggested that addition of salmeteraE to usual asthma therapy is associated with a significant increase in the incidence of serious adverse events and asthma-related deaths compared with addition of albuterol or placebo to usual therapy in the same type of patients. These results prompted the United States Food and Drug Administration (FDA) to issue a stern warning regarding these potential adverse effects and to advise that long-acting beta-agonists (LABAs) should not used as first-line therapy for the treatment of asthma. Two potential explanations have been proposed for these unexpected adverse effects. It has been suggested that more than a direct pharmacological effect of LABAs, these adverse events result from inadequate concomitant use of inhaled corticosteroids in subjects treated with these medicines. However, a detailed analysis of the results of the two large trials did not provide definitive conclusions regarding the potential protective role of inhaled corticosteroids. A second explanation, which the author here considers more plausible, is that a small group of patients with asthma develop idiosyncratic responses to LABAs, and common or rare variants in the genes that encode for proteins associated with the pharmacological response to these medicines are strongly suspected to predispose for these unusual deleterious responses. Until the biological mechanisms involved are better understood, efforts should be made to confine the use of LABAs to those patients who really need them. PMID- 17085800 TI - Concluding remarks: can we explain the association of beta-agonists with asthma mortality? A hypothesis. AB - Beta-agonists have clearly demonstrated benefits for the treatment of both acute and chronic asthma. Therefore, it is perhaps surprising that many of the articles in this issue have focused on concerns about their safety. Much of this concern can be traced back to the "beta-agonist controversy"--the association of high dose isoprenaline and fenoterol inhalers with asthma mortality in the 1960s and 1970s. Although a causal link was never proven, lingering doubts about the safety of beta-agonists remain. It is unclear whether a similar adverse effect is responsible for recently reported association of long-acting beta-agonists with asthma deaths. No mechanism for the beta-agonist controversy was established, but the evidence presented in this collection of articles points to a number of contributing factors. I suggest that a combination of these effects provides a plausible mechanism for the association of frequent beta-agonist use with asthma mortality. Rebound bronchoconstriction and bronchial hyperresponsiveness occur on withdrawal of regular beta-agonist treatment. Regular use of fenoterol is associated with a reduction in morning peak flow suggesting that the overnight interval between doses is sufficient to allow rebound bronchoconstriction. This has not been observed with terbutaline or salbutamol, although rebound phenomena do occur when these drugs are withdrawn for slightly longer periods. Regular use of beta-agonists also leads to tolerance to their bronchoprotective and bronchodilator effects. Tolerance becomes more apparent with worsening bronchoconstriction. In severe asthma, this could result in a poor response to emergency treatment. The combination of rebound deterioration of asthma and a poor response to beta-agonist treatment resulting from tolerance could explain the increased mortality associated with fenoterol and isoprenaline. Both effects are probably caused by downregulation of beta-receptors which occurs with all beta-agonists. Long-acing beta-agonists cause a similar degree of tolerance to short-acting beta-agonists, but avoid the problem of overnight withdrawal. Long acting beta-agonists have also been shown to improve asthma control when taken in combination with inhaled corticosteroids. The clinical significance of tolerance in this context remains to be determined. PMID- 17085801 TI - Microarray analysis of cardiovascular diseases. AB - Microarray analysis is a powerful technique for high-throughput, global transcriptonomic profiling of gene expression. It holds great promise for analyzing the genetic and molecular bases of cardiovascular diseases and various other complex diseases and permits the analysis of thousands of genes simultaneously, both in diseased and nondiseased tissues and/or cell lines. Microarrays or microchips are made by depositing spots of DNA or oligonucleotides representing thousands of genes on a solid support such as a coated glass surface, and can allow the comparison of gene expression patterns in any two samples. Total RNA is isolated from the tissue or cells of interest, converted to cDNA and then cRNA labeled with biotin, and hybridized to the chips. Hybridization signals are then quantified and compared among different samples. We used oligonucleotide microarrays to obtain an unbiased assessment of expression levels of thousands of genes simultaneously in normal and diseased coronary arteries. Fifty-six genes showed differential expression in atherosclerotic coronary artery tissues, and 49 of them represent new linked genes for coronary artery disease. These studies can generate novel hypotheses relating to the pathologies of disease and further studies with animal models, molecular biology, cell biology, and biochemistry will validate these hypotheses and provide novel insights into the pathogenesis of disease. PMID- 17085802 TI - Proteomics with two-dimensional gel electrophoresis and mass spectrometry analysis in cardiovascular research. AB - Proteomics is a large-scale, comprehensive study of the proteins of a cell or organism. It is a unique means of characterizing proteins that are expressed in a cell or tissue at any given time-point and of identifying any modifications that they may undergo. Thus, it is a powerful technology that can detect and identify the changes of the structure and function of proteins in response to intra- and extracellular environmental signals or disease states. As proteomics can establish a link for genes and proteins with a disease, it will play an important role in defining the molecular determinants of a disease and in identifying targets for drug discoveries and diagnostics. We have carried out the first proteomics study for coronary artery disease (CAD) and found that the expression of the ferritin light chain was significantly increased in CAD tissues. In this chapter, we use the CAD study as an example to demonstrate the procedures involved in proteomics analysis. The proteome is visualized by two-dimensional gel electrophoresis, a powerful and widely used method for proteomics, and the proteins of interest are then identified by mass spectrometry. This technique should be useful in characterizing cardiovascular diseases and in defining signaling pathways for cardiovascular development and physiology. PMID- 17085803 TI - Developing and evaluating genomics- or proteomics-based diagnostic tests: statistical perspectives. AB - The completion of the Human Genome Project and the ongoing sequencing of mouse, rat, and other genomes have led to an explosion of genetics-related technologies that are finding their way into all areas of biological research in both basic sciences and clinical applications. High-throughput genomics and proteomics technology has been quickly adapted to develop tools for clinical and pharmacological applications. Because molecular alterations usually occur much earlier than histological, physiological, and clinical abnormality, researchers hope to extend the applications of genomics and/or proteomics technology to early diagnosis of diseases and clinical outcome prognosis. Recently, some successful attempts in molecular diagnosis or prognosis have been published. However, for such tests to be translated from the bench to the bed, they must meet some rigorous standards. To develop a clinically meaningful genomics-based diagnostic test, we must have good study design, appropriate statistical analyses, and valid assessment of its clinical efficacy. In this chapter, we discuss statistical considerations on the process of developing reliable and useful genomics- or proteomics-based tests. PMID- 17085804 TI - Animal models for disease: knockout, knock-in, and conditional mutant mice. AB - Diseases with a genetic basis can be modeled with knockout, knock-in, and conditional mutant gene-targeted mice. In the following, we provide detailed protocols for gene targeting. Gene targeting of embryonic stem cells can be accomplished by laboratories equipped for tissue culture. Alternatively, many gene-targeting services divide the work of targeting with a customer lab. In this collaborative situation, knowledge of the entire process helps ensure a successful outcome. The construction of chimeras for germ-line transmission is not described here, because this procedure is beyond the means of most laboratories, typically is provided by transgenic core facilities, and is best learned through hands-on demonstration. PMID- 17085805 TI - Generation of transgenic mice for cardiovascular research. AB - The transgenic mouse technology is a powerful tool that can be used for creating animal models for cardiovascular disease to identify molecular pathogenic mechanisms and for identifying the physiological functions of a novel gene. A transgenic animal can be generated by several methods, which include microinjection of a DNA fragment into the pronucleus, embryonic stem cell manipulation and injection, sperm-mediated transgenesis, and viral infection of preimplanted embryos. The microinjection method is one of the most widely used approaches. This method involves four steps: (1) collection of fertilized eggs from the superovulated female, (2) injection of DNA into the pronucleus of fertilized eggs, (3) transfer of the injected eggs back into the oviduct of a pseudopregnant foster recipient, allowing the eggs to develop into pups, and (4) identification of the transgenic founder and establishment of transgenic lines through further breeding. PMID- 17085806 TI - Quantitative assay for mouse atherosclerosis in the aortic root. AB - The mouse has become the preferred species for genetic manipulation aimed at creating and studying models for human disease. Although mice are highly resistant to atherosclerosis, dietary induction and, more frequently, gene knockout and transgenic mice have been widely used to study factors that alter the susceptibility to atherosclerosis. Although there are several ways to assess atherosclerosis in mice, measurement of the aortic root lesion area is a commonly used, medium-throughput method that allows for histological examination of the lesions. Here, we provide the detailed methods for the quantitative analysis of mouse aortic root lesion area. PMID- 17085807 TI - Animal models for heart failure. AB - Heart failure (HF) is a major cause of morbidity and mortality worldwide. Although many therapeutic means are available to prolong the life of HF patients, why HF develops is still poorly understood. Investigators still seek a truly appropriate animal model that will reliably mimic human HF, so that the cause of the disease can be targeted and proper therapeutic modalities implemented. HF is a complex condition in which multiple molecular mechanisms interact, resulting in compromised cardiac function and often death. Once this elusive animal model is found, investigators will be able to translate findings from the model to human disease, thereby allowing analysis of the molecular changes and dissecting out multiple complicated changes in HF cascade. In this chapter, we describe the methodology that is used to analyze both transcriptional and translational molecular changes and correlate them with cardiac function to assess the cause and-effect relationship to HF. We used one particular animal model of HF as an example (induced by causing overexpression of myotrophin specifically in the heart) that allowed us to analyze the changes during initiation, progression, and transition of hypertrophy to HF. We have also summarized some other animal models of HF currently available to study mechanisms of HF. PMID- 17085808 TI - Animal models for hypertension/blood pressure recording. AB - Hypertension is an important disease with polygenic inheritance. In order to identify the genes involved in blood pressure regulation, hypertensive rat and mouse models have been developed either by selective breeding or by transgenic technology. The most essential technological prerequisite in these studies is a reliable assessment of the blood pressure in rodents. Three methods are used most frequently for this purpose: tail cuff plethysmography, intra-arterial catheters, and radiotelemetry. Plethysmography is noninvasive, relatively simple, and suitable for a large number of animals, but also imprecise. Intra-arterial catheters are more precise, but require surgery. And both methods restrain and thereby stress the animals, which leads to alterations in blood pressure. Therefore, the telemetric blood pressure measurement, which allows the study of conscious, freely moving animals, has become the gold standard for measuring blood pressure in rodents. However, this method is extremely expensive. Thus, for each experiment the costs have to be put in relation to the quality of data required. This chapter will describe blood pressure measurement methods in technical detail. PMID- 17085809 TI - Animal models for cardiac arrhythmias. AB - Transgenic and gene-targeted mice are now frequently used to expand the study of cardiac physiology and pathophysiology owing to the ease with which the mouse genome can be manipulated. There are many measures by which an assessment of the phenotypical expression of the transgenic mouse can be made. In the case of cardiac channelopathies and how they relate to cardiac function, telemetry is a technology that utilizes transmitters that are surgically implanted in animals for the purpose of acquiring biopotentials or physiological parameters. Electrophysiological techniques have also been used to assess cardiac function at the cellular level, by measuring whole-cell ionic currents and/or transmembrane potentials. This chapter will discuss the surgical procedures involved in successfully implanting the transmitter device in a mouse, as well as highlight the recording of and analysis of electrocardiograms. This chapter will also outline the procedures involved in isolating single-ventricular myocytes from a mouse heart. It is a protocol that was developed in our laboratory for which we have routinely and successfully isolated myocytes from both transgenic and nontransgenic mouse hearts. Although no one isolation protocol is alike, we also present our own observations that have assisted in maximizing myocyte bioavailability and yield. PMID- 17085810 TI - Optical mapping of shock-induced arrhythmogenesis in the rabbit heart with healed myocardial infarction: fluorescent imaging with a photodiode array. AB - Optical mapping of electrical activity in the heart employs digital imaging and voltage-sensitive dyes. These methods have become an increasingly common research tools in basic cardiac electrophysiology. Significant advantages of this approach include simultaneous noncontact recording of entire action potentials free of electrical stimulus-induced artifacts from multiple closely adjacent sites, and adjustable spatial and temporal resolutions. In this way, the activation pattern as well as the repolarization pattern can be monitored by dynamic registration of transmembrane potential changes. As a result, the success of these techniques is most evident in the investigation of the mechanisms of pacing, vulnerability, and defibrillation, in which conventional electrical recordings are hampered by stimulus-induced artifacts. Using optical mapping technology and instrumentation driven by LabVIEW software, we mapped changes in transmembrane voltage during defibrillation shocks and identified the mechanisms of vulnerability and defibrillation in rabbit hearts with healed myocardial infarction (>or=4 wk postinfarction). PMID- 17085811 TI - Methods for studying voltage-gated sodium channels in heterologous expression systems. AB - The discovery that oocytes of the frog Xenopus laevis can be induced to express working membrane ion channels by introducing channel mRNA into their cytoplasm (heterologous expression) has greatly impacted the field of ion channel physiology. With the addition of site-directed mutagenesis techniques, the functional consequences of virtually any mutation can now be specifically and easily assessed. Here, we describe an effective procedure for investigating cardiac sodium channel gating (hNaV1.5) both in Xenopus oocytes, and in a mammalian expression system, human embryonic kidney (HEK) 293 cells. We describe cell attached patch clamp for oocytes, and whole cell voltage clamp in HEK 293 cells. PMID- 17085812 TI - Laser-induced thrombosis in zebrafish larvae: a novel genetic screening method for thrombosis. AB - Classical genetic approaches to study hemostasis and thrombosis have not been available until our recent introduction of the teleost, Danio rerio (the zebrafish), as an effective genetic model for in vivo coagulation assays. The genetic screen for this model is carried out using the genome saturation mutagenesis approach. The resulting mutants are screened for hemostatic or thrombotic defects. We developed a global physiological screening method for thrombosis by utilizing a laser to induce thrombosis in a specifically targeted area of the major artery and vein. Using this assay, we have screened many fish for abnormal hemostasis, and have isolated a number of mutants with abnormal coagulation parameters. These mutants can be grown, bred, and further evaluated for the genetic etiology of their abnormal hemostatic pathways. PMID- 17085813 TI - Methods for isolation of endothelial and smooth muscle cells and in vitro proliferation assays. AB - Angiogenesis, the formation of new blood vessel from pre-existing blood vessel, occurs in a variety of normal and pathological conditions. It is complex morphogenetic process involving the coordinate migration, invasion, and reorganization of several cell types including endothelial cells, pericytes, smooth muscle cells, and stromal fibroblasts. The angiogenic response begins with excess protease secretion to facilitate basement membrane remodeling, proliferation of endothelial cells, and endothelial cell migration to form capillary network and lumen closure. In this chapter, we describe the methods to isolate mouse and human endothelial cells, smooth muscle cells, which will provide abundant, convenient and useful tool for the investigation of many aspects of endothelial cell biology. The high degrees of functional diversity have been observed from endothelial cells derived from different organs, and within the different vascular beds of a given organ. Therefore, this apparent heterogeneity has highlighted the requirement for the endothelial cell isolation and culture from a variety of tissues of different species in order to establish more realistic in vitro angiogenic models. PMID- 17085814 TI - Applications of adenoviral vector-mediated gene transfer in cardiovascular research. AB - Cardiovascular disease is a leading cause of morbidity and mortality worldwide. New studies are needed to explore novel therapeutic options for patients that are refractory to existing therapies. Gene transfer using adenoviral vectors has shown promising results in animal studies, and is now being tested in many clinical trials. In this chapter, the advantages of adenoviral vector-mediated gene transfer for cardiovascular disease applications, and the methods on how to construct, propagate, and evaluate adenoviral vectors, are discussed. PMID- 17085815 TI - Murine and HIV-based retroviral vectors for in vitro and in vivo gene transfer. AB - The success of experimental gene therapy is dependent on the ability to safely and efficiently introduce transgenes into the target cell or tissue. Retroviral based vectors, notably those derived from Moloney murine leukemia virus (MLV) and lentiviral vectors derived from HIV, have proven to be valuable gene transfer vehicles as a result of their ease of production and their ability to mediate long-term transgene expression. One of the most widely used methods for viral vector production is based on the transient transfection of viral vector plasmid DNA into a producer cell line. Here, we describe protocols to produce and standardize high quality MLV-based retroviral and HIV-based lentiviral vectors for ex vivo and in vivo gene delivery. PMID- 17085816 TI - Efficient transfection of primary cells relevant for cardiovascular research by nucleofection. AB - Cell types that are important for cardiovascular research, e.g., cardiomyocytes, endothelial cells, or adult stem cells, are often hard to isolate, culture, and transfect. Low-transfection efficiencies are a major limitation because, in many cases, results achieved with surrogate model cell lines, if any at all are available for the primary cell type of interest, do not reflect the situation in the primary cell. We have demonstrated that unprecedented transfection results are achieved with primary cells when novel electroporation conditions are combined with a treatment of the cells in specific solutions that help stabilize the cells in the electrical field. This led to the development of the new proprietary transfection technology nucleofection. Nucleofection has proved to be successfully applicable to a variety of primary cells and other hard-to-transfect cell lines, and, thus, opens unique perspectives for novel experimental setups as therapeutic strategies. Herein we present protocols for the efficient nucleofection of human umbilical vein endothelial cells, human coronary artery endothelial cells, smooth muscle cells (e.g., pig vascular smooth muscle cells), neonatal rat cardiomyocytes, and human mesenchymal stem cells and depict some results obtained with such transfected cells. PMID- 17085817 TI - Cell adhesion and migration assays. AB - Adhesion and migration are basic responses of living cells to environmental stimuli. Such responses are central to a broad range of physiological processes, such as the immune response, repair of injured tissues, and prevention of excessive bleeding. Cell adhesion and migration also contributes to pathologies, including vascular and inflammatory diseases, as well as tumor growth and metastasis. These cellular responses depend on engagement of adhesion receptors by components of the extracellular matrix or molecules present on the surface of other cells. Hence, cell adhesion and migration assays are crucial methods in cell biology. In this chapter, several detailed protocols describing cell adhesion and migration assays are presented, and advantages and disadvantages of each method are discussed. PMID- 17085818 TI - Apoptosis assays. AB - A large number of methods devoted to the identification of apoptotic cells and the analysis of the morphological, biochemical, and molecular changes that take place during this universal biological process have been developed. Apoptotic cells are recognized on the basis of their reduced DNA content and morphological changes that include nuclear condensation and which can be detected by flow cytometry (sub-G1 DNA content), Trypan Blue, or Hoechst staining. Changes in plasma membrane composition and function are detected by the appearance of phosphatidylserine on the plasma membrane, which reacts with Annexin V fluorochrome conjugates. Combined with propidium iodide (PI) staining, this method can distinguish between the early and late apoptotic events. The best recognized biochemical hallmarks of apoptosis are the activation of cysteine proteases (caspases), condensation of chromatin, and fragmentation of genomic DNA into nucleosomal fragments. Recognized by a variety of assays, activated caspases cleave many cellular proteins and the resulting fragments may serve as apoptosis markers. Finally, the mitochondria and the Bcl-2 family proteins play an important role in this process that can be recognized by translocation of apoptogenic factors, such as Bax and cytochrome c, in and out of mitochondria. PMID- 17085819 TI - Preparation of protein crystals for X-ray structural study. AB - The knowledge of accurate molecular structures obtained by X-ray protein crystallography is now inevitable for rational drug design and for understanding the molecular basis underlying genetic disorders found in patients. However, preparing protein crystals suitable for structural analysis is currently the bottleneck in structure determination by this method. The intent of this chapter is to present current methods of preparing protein crystals for structural studies for a wide range of biologists who have access to macromolecules but do not know how to handle them for crystallization. The chapter includes the pretreatment of a protein prior to the crystallization experiment, initial screens, and optimization of the crystallization conditions for further X-ray study. Finally, handling considerations that are important for a protein intended for crystallization experiments are discussed. PMID- 17085820 TI - Structural elucidation of integrin alphaIIbbeta3 cytoplasmic domain by nuclear magnetic resonance spectroscopy. AB - Integrin alphaIIbbeta3 is a heterodimeric (alpha/beta) cell surface receptor critical for platelet aggregation, and its dysfunction is linked to thrombosis and a number of other vascular diseases. Upon agonist stimulation, which leads to platelet aggregation, alphaIIbbeta3 is activated via a distinct inside-out signaling pathway, i.e., the short alphaIIb/beta3 cytoplasmic tails receive intracellular signals, which trigger the conformational change of the extracellular domain for the high-affinity ligand binding. The structural basis for how the alphaIIbbeta3 cytoplasmic face regulates the inside-out activation of the receptor has been extensively studied over the past decade. We have recently used nuclear magnetic resonance (NMR) spectroscopy to characterize and determine the structural features of the alphaIIbbeta3 cytoplasmic domain. This chapter describes detailed practical procedures for performing these NMR studies, which have provided key atomic insights into the mechanism of the alphaIIbbeta3 function, especially its inside-out signaling. PMID- 17085821 TI - Applications of electron cryo-microscopy to cardiovascular research. AB - Electron cryo-microscopy (cryo-EM) techniques have wide applications for the study of biological structures. The focus of this chapter is the use of cryo-EM and associated methods for the analysis of the three-dimensional (3D) structure of proteins and multicomponent macromolecular assemblies. Data evolving from these methods pertaining to the quaternary organization of proteins and protein protein interactions, bridges an important gap between linear genomic information and understanding physiological function. This chapter provides general methods for examining two-dimensional crystalline arrays of proteins as well as single, randomly orientated protein complexes. It is significant that single-particle analysis of electron microscopy images has provided the only 3D data to-date for the two principal components of muscle excitation contraction coupling in the heart, namely the L-type voltage-gated calcium channel and the ryanodine receptor. This chapter describes approaches for identifying the extracellular and intracellular domains of the 3D structure of the L-type voltage-gated calcium channel and also incorporates general details for labeling and visualizing His tagged proteins. PMID- 17085822 TI - Stem cells in cardiovascular disease: methods and protocols. AB - Stem cells are cells capable of proliferation, self-renewal, and differentiation into various organ-specific cell types. Stem cells are subclassified based on their species of origin (mice, rat, human), developmental stage of the species (embryonic, fetal, or adult), tissue of origin (hematopoietic, mesenchymal, skeletal, neural), and potential to differentiate into one or more specific types of mature cells (totipotent, pluripotent, multipotent). Embryonic stem (ES) cells are totipotent, primitive cells derived from the embryo that have the potential to become all specialized cell types. Conversely, adult stem cells are undifferentiated cells found in differentiated tissue that retain the potential to renew themselves and differentiate to yield organ-specific tissues. Stem cells are attractive candidates for novel therapeutics for patients with different heart diseases, including congestive heart failure, most commonly caused by myocardial infarction. The remarkable proliferative and differentiation capacity of stem cells promises an almost unlimited supply of specific cell types including viable functioning cardiomyocytes to replace the scarred myocardium following transplantation. PMID- 17085823 TI - Editorial 'autonomy ' in learned journals: lessons from the CMAJ episode. PMID- 17085824 TI - Regulating stem cell research & therapy. PMID- 17085825 TI - Nestin immunoexpression in CNS tumours. PMID- 17085826 TI - Awakening India to obstructive sleep apnoea syndrome. PMID- 17085827 TI - The burden of cardiovascular disease in the Indian subcontinent. AB - Ischaemic heart disease and stroke are among the most common causes of death and disability in the world. The Indian subcontinent (including India, Pakistan, Bangladesh, Sri Lanka, and Nepal) has among the highest rates of cardiovascular disease (CVD) globally. Previous reports have highlighted the high CVD rates among South Asian immigrants living in Western countries, but the enormous CVD burden within the Indian subcontinent itself has been underemphasized. In this review, we discuss the existing data on the prevalence of CVD and its risk factors in the Indian subcontinent. We also review recent evidence indicating that the burden of coronary heart disease in the Indian subcontinent is largely explained on the basis of traditional risk factors, which challenges the common thinking that South Asian ethnicity per se is a strong independent risk factor for coronary heart disease. Finally, we suggest measures to implement in policy, capacity building, and research to address the CVD epidemic in the Indian subcontinent. PMID- 17085828 TI - Adjuvant therapy in cerebral malaria. AB - Cerebral malaria is the most common cause of non-traumatic encephalopathy in the world. The mainstay of therapy is either quinine or artemisinin, both of which are effective antimalarials. The clinical picture of cerebral malaria may persist or even become worse in spite of the clearance of parasites from blood. The death rate is unacceptably high even with effective antimalarials in tertiary care hospitals. The mortality increases in presence of multi organ failure (renal failure, jaundice, respiratory distress, severe anaemia, lactic acidosis, etc.). The pathogenesis of cerebral malaria is multifactorial and includes clogging, sequestration, rosette formation, release of cytokines, cerebral oedema, increased intracranial hypertension, etc. Attempts are made to use adjuvant therapy which will act through alternate mechanisms and address one or more of the pathogenetic processes. In this review, we have discussed the role of corticosteroids, pentoxifylline, desferrioxamine, mannitol and newer agents in the treatment of cerebral malaria. Though the literature on adjuvant therapy in cerebral malaria is large enough, there are a number of shortcomings in the clinical trials, many being open and non randomized or of very small sample size. Further research is of utmost importance through large multicentric, double-blind controlled trials to show the efficacy of any of these drugs. PMID- 17085829 TI - Effect of tobacco smoking on renal function. AB - Nicotine is one of many substances that may be acquired through active and passive smoking of tobacco. In man, nicotine is commonly consumed via smoking cigarettes, cigars or pipes. The addictive liability and pharmacological effects of smoking are primarily mediated by the major tobacco alkaloid nicotine. High stress jobs favour repeated smoking and further reinforce addictive behaviours. There are elevated serum cadmium and lead levels in smokers resulting in glomerular dysfunction. Nephropathies are accelerated by nicotine with an increased incidence of microalbuminuria progressing to proteinuria, followed by type-1 diabetes mellitus induced renal failure. Cigarette smoke-induced renal damage is due, at least in part, to activation of the sympathetic nervous system resulting in an elevation in blood pressure. Ethanol, nicotine, or concurrent intake significantly increases lipid peroxidation in liver, and decreased superoxide dismutase activity and increased catalase activity in the kidney. This review describes the effects of nicotine, smoking, smoke extracts and other tobacco constituents on renal and cardiovascular functions, and associated effects on the nervous system. Both active and passive smoking is toxic to renal function. PMID- 17085830 TI - Expression of nestin--a stem cell associated intermediate filament in human CNS tumours. AB - BACKGROUND & OBJECTIVES: Nestin is an intermediate filament protein expressed in undifferentiated cells during the development of brain and is considered as a marker for neuroepithelial stem cells. Expression of this protein in various CNS tumour cells suggests the possibility of existence of tumour stem cell modulating the evolution. We carried out an immunohistochemical study to demonstrate the expression of nestin and its co-expression with neuronal and glial intermediate filament and correlate with the grade of malignancy. METHODS: Formalin fixed, paraffin processed sections from two human foetuses, 16 brain tumours of both neuronal and glial lineage and two metastatic tumours were immunostained with polyclonal antibody to nestin. Serial sections from primary brain tumours were also stained with monoclonal antibody to neurofilament (NF) and glial fibrillary acidic protein (GFAP). Fluorescent double labeling was carried out on four cases using laser confocal microscopy, to document co-localization of nestin with other intermediate filaments in the tumour cells. RESULTS: Nestin expression was observed along the paraventricular zone of human foetuses and in brain tumours of both glial and neuronal lineage, of both high and low grades of malignancy. In addition, mature dysplastic spinal motor neurons adjacent to tumour and cerebellar Purkinje cells also expressed nestin along with neurofilament. INTERPRETATION & CONCLUSION: Nestin expression was noted in both low and high grade brain tumours and dysplastic neurons and did not parallel the malignant grade of the tumour. The expression of nestin in tumour cells and dysplastic neurons suggests aberrant expression of antigenically primitive proteins in cells to facilitate remodelling of the cell and migration. More studies are needed to elucidate the concept. PMID- 17085831 TI - Validation of the modified Berlin questionnaire to identify patients at risk for the obstructive sleep apnoea syndrome. AB - BACKGROUND & OBJECTIVES: Awareness regarding obstructive sleep apnoea (OSA) among general public as well as practicing physicians is low in India. The present study was undertaken to test the utility of modified Berlin questionnaire for risk categorization of OSA in Indian setting. METHODS: The modified Berlin questionnaire was administered in 180 middle aged adults (of 320 screened), of whom, 104 underwent overnight polysomnograhy, in a cross-sectional study at a tertiary care, referral center in north India. Questionnaire addressed the presence of frequency of snoring, wake time sleepiness, fatigue, obesity and hypertension. Subjects with persistent and frequent symptoms in any two of these three domains were considered in high risk category for obstructive sleep apnoea. Overnight polysomnograhy was performed to measure apnoea and hypopnoea index (AHI). RESULTS: Questions about the symptoms demonstrated internal consistency (Cronbach alpha correlations 0.92-0.96). Of the 180 respondents to the screening questions, 80 were in the high risk and the rest were in low risk group. For 104 subjects who underwent polysomnograhy, risk grouping was useful in prediction of AHI. High risk category predicted an AHI >5 with a sensitivity of 86 per cent, specificity of 95 per cent, positive and negative predictive values of 96 and 82 per cent respectively. These results were comparable to Berlin questionnaire study done in the western population for validation. INTERPRETATION & CONCLUSION: On the basis of the findings of present study it is concluded that administration of modified Berlin questionnaire prior to a polysomnography study can identify high risk subjects and can thus avoid unnecessary polysomnography studies especially in resource-limited settings. To identify subjects at risk for OSA syndrome in general population, this questionnaire can be applied. However, the findings of the present study need to be confirmed further in a large number of subjects in a community-based setting. PMID- 17085832 TI - Frequency of major affective disorders in first-degree relatives of patients with type 2 diabetes mellitus. AB - BACKGROUND & OBJECTIVES: A genetic link between diabetes and depression has been proposed, but hardly explored. Data on family studies exploring relation between depression and diabetes are scanty. This study attempted to assess the prevalence of major affective disorders in first-degree relatives of patients with type 2 diabetes mellitus (T 2 DM). METHODS: Fifty probands with T 2 DM, in whom other psychiatric disorders had been excluded, were chosen. Morbid risks and prevalence figures for depression and mania were estimated in 481 first-degree relatives of these 50 probands using the family interview for genetic studies. RESULTS: Of the 481 first-degree relatives of probands, only six had affective disorders. The morbid risk for depression in first-degree relatives was 2.99 and 3.87 per cent, assuming age of risk at 15-60 and 15-50 yr respectively, while the morbid risk for mania was 0.59 and 0.77 per cent in these age groups. INTERPRETATION & CONCLUSION: The morbid risks/prevalence rates among first-degree relatives of probands with T2 DM were not higher than those of the general population rates derived from earlier Indian and western studies. This study did not demonstrate a family aggregation of affective disorders in patients with T 2 DM. Increased prevalence of affective disorders in diabetes could be due to non-genetic factors. PMID- 17085833 TI - Hormonal abnormalities in azoospermic men in Kano, Northern Nigeria. AB - BACKGROUND & OBJECTIVES: We undertook this study to observe the pattern of hormonal abnormalities and testicular pathology in azoospermic male Africans in Kano, Northern Nigeria. METHODS: Eighty consecutive azoospermic infertile males attending fertility clinic in Aminu Kano Teaching Hospital, Kano, were selected for the study. Their semen were analyzed three times at eight weeks interval, after which serum follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone and prolactin were assayed in serum samples, and histological examination of testicular biopsies done. RESULTS: Of the 80 subjects studied, 32 (40%) had abnormal hormonal levels, 48 (60%) had normal hormonal values and 36 (45%) had testicular pathology. INTERPRETATION & CONCLUSION: Endocrinopathies are common in azoospermia. Their contribution to male factor infertility cannot be overemphasized. The main reason for the endocrinopathies is not known but environmental factors, endocrine disruptors and genetic polymorphism have been suggested to be contributory. PMID- 17085834 TI - HLA alleles in pre-menopausal breast cancer patients from western India. AB - BACKGROUND & OBJECTIVES: Cytotoxic function of Natural Killer (NK) cells is regulated by the products of HLA class I genes. Associations between HLA alleles and risk for cancers have been suggested earlier. Several reports further emphasize the need to examine influence of HLA genotypes on risk for malignant disorders. Therefore, we undertook this study to assess the possibility of association of HLA-class I alleles in pre-menopausal breast cancer patients. METHODS: Eighty one pre-menopausal breast cancer patients and 160, age and ethnicity matched healthy women from western India were studied. Genotyping for HLA class I alleles and HLA-B*40 alleles (high resolution) was performed using genotyping kits from Genovision Inc., USA. RESULTS: Nearly two-fold higher frequency of HLA - B*40 (16%) was observed in patients compared to controls (O.R. = 2.2; 95% C.I.-1.15-4.34; P<0.02). High resolution typing for HLA-B*40 alleles revealed predominance of HLA-B*4006 allele in both the study groups. Two other important observations relate to lower frequency of HLA - B*08 in patients and homozygosity at HLA-Cw locus in significantly higher proportion of patients. INTERPRETATION & CONCLUSION: The nature of peptides presented by HLA-B*4006 may have implications for higher frequency of HLA-B*40 in breast cancer patients. Higher frequency of homozygosity at HLA-Cw alleles in patients suggests a role for killer immunoglobulin-like receptors (KIRs) in NK cell mediated immune surveillance. Results of this study provide directions to further analyse role of immunogenetic mechanisms governing innate and adaptive immune responses contributing to modulation of risk for breast cancer. PMID- 17085835 TI - Intestinal enzymes during malnutrition & infection in rabbits. AB - BACKGROUND & OBJECTIVES: Malnutrition plays an important role in the intestinal absorption of nutrients. However, reports are not consistent whether intestinal enzymes are decreased in the presence of malnutrition. It is also not clear whether simultaneous presence of malnutrition and infection adds to the problem of malabsorption of nutrients. The aim of the present study was to determine intestinal functions in terms of concentrations of disaccharidase enzymes during diarrhoea and protein energy malnutrition. METHODS: Concentrations of three disaccharidase enzymes, namely maltase, sucrase and lactase were measured in nine energy-restricted and five control rabbits during diarrhoea induced by rabbit diarrhoeagenic Escherichia coli (RDEC-1). Malnutrition was achieved in the rabbit model by feeding the animals for 30 days with half the amount of food fed to well nourished control rabbits. Both the energy-restricted and the control groups were challenged by RDEC-1. Diarrhoea occurred on day 1-7 after administration of the strain. After onset of diarrhoea, both groups of rabbits were sacrificed and their intestinal mucosa was examined to determine the concentration of lactase, maltase and sucrase. RESULTS: The energy-restricted animals and controls did not differ significantly for concentrations (units/mg proteins) of lactase (0.65 +/- 0.28 vs 0.56 +/- 0.17 ), maltase (6.20 +/- 2.70 vs 6.47 +/- 1.90) and sucrase (5.42 +/- 2.30 vs 5.13 +/- 1.40) measured during acute infectious diarrhoea. INTERPRETATION & CONCLUSION: The results suggested that the enzymatic functions of the intestinal brush border were not statistically different during diarrhoea among malnourished rabbits compared with their well-nourished counterparts. PMID- 17085836 TI - CD4/CD8 lymphocyte counts in healthy, HIV-positive individuals & AIDS patients. AB - BACKGROUND & OBJECTIVES: The enumeration of CD4 and CD8 positive cells, surrogate markers for HIV disease progression, is helpful in management and follow up of immunocompromised HIV-positive patients. In assessing the degree of immune deficiency in HIV-positive patients of a particular region, knowledge of reference range of T-cell subset counts of healthy individuals of that particular region is essential. The present cross-sectional study was undertaken to determine the reference range of T-cell subsets in healthy north Indians and to compare the values with those in HIV-positives. METHODS: Blood samples from 125 HIV seronegative healthy volunteers comprising group I (88 males, 37 females) and 452 HIV- positive patients, divided into group II of asymptomatic (n=137; 93 males, 44 females) and group III of AIDS patients (n=315; 253 males, 62 females) in the age group of 17-60 yr, were analysed for enumeration of CD4+, CD8+ cells/microl by flow cytometry. RESULTS: In group I, the CD4 and CD8 levels were 687 +/- 219 and 611 +/- 288 cells/microl in males and 740 +/- 255 and 546 +/- 246 cells/microl in females. Overall, a significant depressed level of CD4 (525 +/- 207 cells/microl) and elevated level of CD8 (1174 +/- 484 cells/microl) in group II and (170 +/- 115 and 1051 +/- 586 cells/microl) respectively in group III were observed. Group II patients had highest level of CD8 cells. No asymptomatic women had CD4 count of <200 cells/microl. INTERPRETATION & CONCLUSION: Our findings on T-cell subset reference ranges of normal healthy north Indians validate the utility of determination of CD4 cell count as a useful predictor of AIDS in Indian conditions and confirm that a significant per cent of AIDS patients had CD4 cell count below 200/microl. PMID- 17085837 TI - Kindling & mossy fibre sprouting in the rat hippocampus following hot water induced hyperthermic seizures. AB - BACKGROUND & OBJECTIVES: Hot water epilepsy (HWE) is well recognized reflex epilepsy with possible genetic susceptibility. Rat model and human experimentation had proven that HWE is a type of hyperthermic seizure with possible kindling on repeated stimulation in animals. The present study was undertaken to investigate kindling associated with hyperthermic seizures induced by repeated hot water stimulation in the rat model and to prove hyperthermic kindling. METHODS: Epileptic seizures were induced in 36 male Wistar albino rats by means of hot water sprays at 48 h time intervals. Progression of seizure activity was investigated by studying the behaviour, severity and duration of the seizure. Threshold of rectal temperatures and timed latency for seizure induction were studied. Seizure discharges (EEG) were recorded from ventral hippocampus in six of these rats. Timm's staining was used to study the neuronal sprouting as a consequence of kindling. Studying the seizure threshold, latency, duration of seizure discharge and behavioural seizure following a stimulus-free interval of 30 days tested permanence of kindling. RESULTS: Following 8-12 episodes of hot water stimulations there was progressive epileptic activity manifested in the form of lowering of rectal temperature thresholds from 41.5 to 40.0 degrees C, drop in latency for developing seizures from 185 to 118 sec, increase in duration of hippocampal seizure discharge from 15 to 140 sec, along with progressive increase in complexity of EEG after discharges, increase in behavioural seizure severity from Grade 1 to 5 in all the rats, and neuronal sprouting observed in supragranular molecular layer and in stratum lacunosum. INTERPRETATION & CONCLUSION: Our study covered all aspects of kindling and provided a useful animal model for human hot water epilepsy. Hyperthermic seizures induced by hot water in the rat model kindle as demonstrated by Timm's staining. PMID- 17085838 TI - Recombinant human erythropoietin (rhEPO) preconditioning on nuclear factor-kappa B (NF-kB) activation & proinflammatory cytokines induced by myocardial ischaemia reperfusion. AB - BACKGROUND & OBJECTIVES: Erythropoietin (EPO), originally identified for its critical hormonal role in promoting erythrocyte survival and differentiation, has shown to a protective effect in myocardial ischaemia-reperfusion (I-R) injury in animal model. However, the precise mechanisms remain unclear. The objective of this study was to determine the roles of nuclear factor-kappa B (NF-kB) and associated cytokines induced by I-R in the cardioprotection by recombinant human erythropoietin (rhEPO). Morphopathological observations were also made on the ultrastructure of myocardial tissue. METHODS: Myocardial I-R rat model was established by 30 min ligation of left descending coronary and 3 h reperfusion. RhEPO or saline solution was intraperitoneally injected 24 h before I-R insult. The infarct sizes were measured by triphenyltetrazolium chloride (TTC)-Evans blue technique and ultrastructural organizations were observed by a transmission electron microscope. Tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL 6), and IL-10 concentrations were analyzed by enzyme-linked immunosorbance assays and NF-kB by electrophoretic mobility shift assay. TNF-alpha and IL-6 mRNA expression were studied by the reverse-transcription polymerase chain reaction (RT-PCR). RESULTS: A single bolus injection of 5,000 units/kg of rhEPO 24 h before insult remarkably reduced infarct size and improved ultrastructural organization of I-R myocardium. It greatly suppressed TNF-alpha and IL-6 expression, but enhanced IL-10 production. It modestly activated NF-kB before I-R insult and markedly attenuated subsequent NF-kB activation during sustained I-R. INTERPRETATION & CONCLUSION: The suppression of proinflammatory cytokines expression may act by inhibiting NF-kB activation during I-R, but not by induction of IL-10. This might be one of the molecular mechanisms of rhEPO in cardioprotection. In addition, NF-kB was suggested to play a dual role in cardioprotective effects of rhEPO. PMID- 17085839 TI - Development of an immunodetection test for a botulinum-like neurotoxin produced by Clostridium sp. RKD. AB - BACKGROUND & OBJECTIVES: Clostridial neurotoxins are among the most toxic substances known and cause severe illnesses in both humans and animals. A neurotoxigenic Clostridium sp. (strain RKD) isolated from intestine of decaying fish produced a novel, botulinum type B like neurotoxin as suggested by mouse bioassay, protection with anti-botulinum antibodies and PCR. The aim of the present investigation was to develop a laboratory based detection assay as an alternative to the mouse bioassay without compromising sensitivity and specificity. METHODS: Growth and toxin production were carried out in trypticase peptone yeast-extract glucose (TPYG) broth. Toxicity was estimated in terms of minimum lethal dose (MLD) by mouse bioassay. The toxin was partially purified by acid precipitation. It was used for toxoid preparation by formaldehyde treatment. This purified IgG was used for detection of neurotoxin using indirect ELISA. The culture supernatant was concentrated using a stirred cell with a 50 kDa cut-off membrane at 4 degrees C. Further purification was carried out using Prep cell. Fractions showing toxicity and sufficient purity were pooled, concentrated and analyzed on sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: The toxin was purified with a recovery of 8.56 per cent. Polyclonal antiserum was raised in mice using partially purified toxin with a titre of 1: 80000. A detection assay with sensitivity of approximately 15 and 300 ng/ml for partially purified and crude toxins, respectively were achieved using an indirect ELISA method. INTERPRETATION & CONCLUSION: The Clostridium sp. RKD produced a potent neurotoxin earlier shown to have novelties. A specific detection assay for the neurotoxin has been developed that may be useful both from food safety and clinical point of view. PMID- 17085840 TI - Nocardia pyopneumothorax in an immunocompetent patient. PMID- 17085842 TI - Using max cut to enhance rooted trees consistency. AB - Supertree methods are used to construct a large tree over a large set of taxa from a set of small trees over overlapping subsets of the complete taxa set. Since accurate reconstruction methods are currently limited to a maximum of a few dozen taxa, the use of a supertree method in order to construct the tree of life is inevitable. Supertree methods are broadly divided according to the input trees: When the input trees are unrooted, the basic reconstruction unit is a quartet tree. In this case, the basic decision problem of whether there exists a tree that agrees with all quartets is NP-complete. On the other hand, when the input trees are rooted, the basic reconstruction unit is a rooted triplet and the above decision problem has a polynomial time algorithm. However, when there is no tree which agrees with all triplets, it would be desirable to find the tree that agrees with the maximum number of triplets. However, this optimization problem was shown to be NP-hard. Current heuristic approaches perform min cut on a graph representing the triplets inconsistency and return a tree that is guaranteed to satisfy some required properties. In this work, we present a different heuristic approach that guarantees the properties provided by the current methods and give experimental evidence that it significantly outperforms currently used methods. This method is based on a divide and conquer approach, where the min cut in the divide step is replaced by a max cut in a variant of the same graph. The latter is achieved by a lightweight semidefinite programming-like heuristic that leads to very fast running times. PMID- 17085843 TI - Pattern identification in biogeography. AB - Identifying common patterns among area cladograms that arise in historical biogeography is an important tool for biogeographical inference. We develop the first rigorous formalization of these pattern-identification problems. We develop metrics to compare area cladograms. We define the maximum agreement area cladogram (MAAC) and we develop efficient algorithms for finding the MAAC of two area cladograms, while showing that it is NP-hard to find the MAAC of several binary area cladograms. We also describe a linear-time algorithm to identify if two area cladograms are identical. PMID- 17085844 TI - Efficient detection of network motifs. AB - Motifs in a given network are small connected subnetworks that occur in significantly higher frequencies than would be expected in random networks. They have recently gathered much attention as a concept to uncover structural design principles of complex networks. Kashtan et al. [Bioinformatics, 2004] proposed a sampling algorithm for performing the computationally challenging task of detecting network motifs. However, among other drawbacks, this algorithm suffers from a sampling bias and scales poorly with increasing subgraph size. Based on a detailed analysis of the previous algorithm, we present a new algorithm for network motif detection which overcomes these drawbacks. Furthermore, we present an efficient new approach for estimating the frequency of subgraphs in random networks that, in contrast to previous approaches, does not require the explicit generation of random networks. Experiments on a testbed of biological networks show our new algorithms to be orders of magnitude faster than previous approaches, allowing for the detection of larger motifs in bigger networks than previously possible and thus facilitating deeper insight into the field. PMID- 17085845 TI - Motif search in graphs: application to metabolic networks. AB - The classic view of metabolism as a collection of metabolic pathways is being questioned with the currently available possibility of studying whole networks. Novel ways of decomposing the network into modules and motifs that could be considered as the building blocks of a network are being suggested. In this work, we introduce a new definition of motif in the context of metabolic networks. Unlike in previous works on (other) biochemical networks, this definition is not based only on topological features. We propose instead to use an alternative definition based on the functional nature of the components that form the motif, which we call a reaction motif. After introducing a formal framework motivated by biological considerations, we present complexity results on the problem of searching for all occurrences of a reaction motif in a network and introduce an algorithm that is fast in practice in most situations. We then show an initial application to the study of pathway evolution. Finally, we give some general features of the observed number of occurrences in order to highlight some structural features of metabolic networks. PMID- 17085846 TI - A 1.375-approximation algorithm for sorting by transpositions. AB - Sorting permutations by transpositions is an important problem in genome rearrangements. A transposition is a rearrangement operation in which a segment is cut out of the permutation and pasted in a different location. The complexity of this problem is still open and it has been a 10-year-old open problem to improve the best known 1.5-approximation algorithm. In this paper, we provide a 1.375-approximation algorithm for sorting by transpositions. The algorithm is based on a new upper bound on the diameter of 3-permutations. In addition, we present some new results regarding the transposition diameter: we improve the lower bound for the transposition diameter of the symmetric group and determine the exact transposition diameter of simple permutations. PMID- 17085847 TI - New bounds and tractable instances for the transposition distance. AB - The problem of sorting by transpositions asks for a sequence of adjacent interval exchanges that sorts a permutation and is of the shortest possible length. The distance of the permutation is defined as the length of such a sequence. Despite the apparently intuitive nature of this problem, introduced in 1995 by Bafna and Pevzner, the complexity of both finding an optimal sequence and computing the distance remains open today. In this paper, we establish connections between two different graph representations of permutations, which allows us to compute the distance of a few non-trivial classes of permutations in linear time and space, bypassing the use of any graph structure. By showing that every permutation can be obtained from one of these classes, we prove a new tight upper bound on the transposition distance. Finally, we give improved bounds on some other families of permutations and prove formulas for computing the exact distance of other classes of permutations, again in polynomial time. PMID- 17085848 TI - Comparing tandem repeats with duplications and excisions of variable degree. AB - Traditional sequence comparison by alignment employs a mutation model comprised of two events, substitutions and indels (insertions or deletions) of single positions. However, modern genetic analysis knows a variety of more complex mutation events (e.g., duplications, excisions, and rearrangements), especially regarding DNA. With ever more DNA sequence data becoming available, the need to accurately compare sequences which have clearly undergone more complicated types of mutational processes is becoming critical. Herein we introduce a new method for pairwise alignment and comparison of sequences with respect to the special evolution of tandem repeats: substitutions and indels of single positions and, additionally, duplications and excisions of variable degree (i.e., of one or more repeat copies simultaneously) are taken into account. To evaluate our method, we apply it to the spa VNTR (variable number of tandem repeats) cluster of Staphylococcus aureus, a bacterium of high medical importance. PMID- 17085849 TI - Faster algorithms for optimal multiple sequence alignment based on pairwise comparisons. AB - Multiple Sequence Alignment (MSA) is one of the most fundamental problems in computational molecular biology. The running time of the best known scheme for finding an optimal alignment, based on dynamic programming, increases exponentially with the number of input sequences. Hence, many heuristics were suggested for the problem. We consider a version of the MSA problem where the goal is to find an optimal alignment in which matches are restricted to positions in predefined matching segments. We present several techniques for making the dynamic programming algorithm more efficient, while still finding an optimal solution under these restrictions. We prove that it suffices to find an optimal alignment of the predefined sequence segments, rather than single letters, thereby reducing the input size and thus improving the running time. We also identify "shortcuts" that expedite the dynamic programming scheme. Empirical study shows that, taken together, these observations lead to an improved running time over the basic dynamic programming algorithm by 4 to 12 orders of magnitude, while still obtaining an optimal solution. Under the additional assumption that matches between segments are transitive, we further improve the running time for finding the optimal solution by restricting the search space of the dynamic programming algorithm. PMID- 17085850 TI - Efficient parameterized algorithms for biopolymer structure-sequence alignment. AB - Computational alignment of a biopolymer sequence (e.g., an RNA or a protein) to a structure is an effective approach to predict and search for the structure of new sequences. To identify the structure of remote homologs, the structure-sequence alignment has to consider not only sequence similarity, but also spatially conserved conformations caused by residue interactions and, consequently, is computationally intractable. It is difficult to cope with the inefficiency without compromising alignment accuracy, especially for structure search in genomes or large databases. This paper introduces a novel method and a parameterized algorithm for structure-sequence alignment. Both the structure and the sequence are represented as graphs, where, in general, the graph for a biopolymer structure has a naturally small tree width. The algorithm constructs an optimal alignment by finding in the sequence graph the maximum valued subgraph isomorphic to the structure graph. It has the computational time complexity O[k(t)N(2)] for the structure of N residues and its tree decomposition of width t. Parameter k, small in nature, is determined by a statistical cutoff for the correspondence between the structure and the sequence. This paper demonstrates a successful application of the algorithm to RNA structure search used for noncoding RNA identification. An application to protein threading is also discussed. PMID- 17085851 TI - Interactions of excitatory amino acid receptor antagonists with antiepileptic drugs in three basic models of experimental epilepsy. AB - Successful management of epilepsy still remains a vital problem. Despite using various combinations of antiepileptic drugs (AEDs), 20-25% of epileptic patients are insensitive to currently available medication. Therefore, there is a considerable need for finding a more effective AED or synergistic combinations of AEDs. Experimental and clinical data indicate that excitatory amino acid (EAA) receptor antagonists possess anticonvulsant potential. Moreover, EAA antagonists can potentiate the protective action of conventional AEDs. Unfortunately, not all beneficial (in terms of anticonvulsant activity) combinations may be recommended since some of them produce significant adverse effects which restrict their clinical use. The aim of this review was to assemble current literature data on interactions of EAA receptor antagonists with conventional AEDs. Generally, N methyl-D-aspartate (NMDA) receptor antagonists combined with AEDs produce significant adverse effects. Non-NMDA receptor antagonists represent a more promising group. PMID- 17085852 TI - Prodrugs and soft drugs. AB - This review focuses on a new approach to the development of drugs, namely on prodrugs and soft drugs. Nowadays, we try to design drugs that heal sick people having the best acceptance of patients. They must be efficient and selective on their site of action and must be metabolized to non-toxic derivatives. Both, prodrugs and soft drugs should have good distributive properties to enhance their quality. They are designed to maximize the amount of an active drug that reaches its target, through changing the physicochemical, biopharmaceutical or pharmacokinetic properties of drugs. Prodrugs are changed into the active drug within the body through enzymatic or non-enzymatic reactions. Soft drugs are novel and active analogues of already known therapeutic agents. It is expected that continued studies will improve drug properties so as to achieve the best drug delivery system. PMID- 17085853 TI - Update on the management of polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is a common endocrine disorder that affects 5% to 10% of women in reproductive age. It is the most simply defined as the association of hyperandrogenism (diagnosed clinically and/or biochemically) with chronic anovulation in the absence of specific ovarian, adrenal and pituitary gland diseases. Recent studies have revealed that PCOS is associated with hyperinsulinemia and insulin resistance. Both of them have been shown to play a substantial role in the increased risk of the cardiovascular events. The new data concerning the pathogenesis and management of PCOS have changed its treatment strategy. The management of PCOS is complex and includes lifestyle modification combined with dietary-induced weight loss, oral contraceptives, clomiphene citrate, gonadotropins, antiandrogens and insulin-sensitizing agents. Women with PCOS, diagnosed and managed properly, can benefit from the reduction or even reversal of the reproductive and metabolic morbidities and from the reduction of the risk factors for cardiovascular disorders. In this article, we describe all these treatment options. Although both standard and novel therapies are discussed, this review focuses on the progress made in the recent years. PMID- 17085854 TI - Effect of BD 1047, a sigma1 receptor antagonist, in the animal models predictive of antipsychotic activity. AB - The sigma receptors were first classified as a subtype of opioid receptors but later they were found to be a distinct pharmacological entity. Many preclinical and clinical data have indicated that sigma receptor ligands have to be involved in neuropsychiatric disorders, including schizophrenia. Numerous data have suggested that potential antipsychotic activity of sigma ligands results from their "antagonistic" activity. However, the subcellular mechanisms by which sigma ligands exert their effects have not been elucidated in detail, therefore, the terms "agonist" or "antagonist" and their functional implications are not entirely unequivocal. The aim of the present study was to find out if BD 1047, described recently as a selective functional antagonist of sigma receptors, shows antipsychotic activity in animal models predictive of efficacy in schizophrenia. In contrast to rimcazole and panamesine, two selective sigma ligands whose antipsychotic activity was confirmed clinically, BD 1047 did not decrease amphetamine-induced hyperactivity in mice in a statistically significant manner. Likewise, it did not modify the hyperactivity induced by NMDA receptor antagonists, phencyclidine, memantine or dizocilpine. On the other hand, BD 1047 attenuated apomorphine-induced climbing in mice and phencyclidine-induced head twitches in rats, like rimcazole and panamesine did. Summing up, BD 1047 shows a moderate activity in models used in this study suggesting that its usefulness as an antipsychtic drug is doubtful. However, more detailed studies are required for definitive confirmation of this conclusion. PMID- 17085855 TI - Inhibition of restraint-induced neuroendocrine and serotonergic responses by buspirone in rats. AB - The effects of buspirone (0.5 mg/kg) on the neuroendocrine and serotonergic responses to stress were monitored in rats. Exposure to 2-h of restraint stress increased circulating levels of corticosterone, noradrenaline and glucose. The metabolism of 5-hydroxytryptamine (5-HT; serotonin) increased in the brain. Prior administration of buspirone did not alter levels of corticosterone, noradrenaline and glucose in unrestrained rats, but inhibited stress-induced increase in the activity of hypothalamic-pituitary-adrenal (HPA) axis and circulating levels of glucose. Restraint-induced rise in brain 5-HT and 5-hydroxyindole-acetic acid (5 HIAA) was also attenuated by buspirone. Unrestrained animals injected with buspirone also exhibited a decrease in brain 5-HIAA concentration. The findings are discussed in the context of the role of somatodendritic 5-HT(1A) receptors in responses to stress. PMID- 17085856 TI - Adenosine receptor antagonists intensify the benzodiazepine withdrawal signs in mice. AB - The aim of the present experiment was to assess the involvement of adenosine receptor antagonists in benzodiazepine (BDZ) withdrawal signs, observed as the seizure susceptibility in mice. The discontinuation of chronic treatment with temazepam or diazepam decreased seizure threshold (one of BDZ withdrawal signs). The concomitant application of subconvulsive dose of pentetrazole (55.0 mg/kg) with low dose of flumazenil (5.0 mg/kg) - a BDZ receptor antagonist, immediately induced BDZ withdrawal signs in these animals. The non-selective adenosine receptor antagonist (caffeine), and the selective adenosine A1 receptor antagonist (DPCPX), injected 15 min before the application of pentetrazole and flumazenil, were able to intensify BDZ withdrawal signs in mice. The most apparent effects were observed after administration of DPCPX, indicating that the adenosine A1 receptor may play a more important role in these effects. The obtained data demonstrate that the adenosinergic system is involved in BDZ withdrawal signs in mice, and adenosine A1 receptor plays an important role in this process. PMID- 17085857 TI - Acute exposure to caffeine decreases the anticonvulsant action of ethosuximide, but not that of clonazepam, phenobarbital and valproate against pentetrazole induced seizures in mice. AB - This study examines the effect of acute administration of caffeine sodium benzoate (CAF) on the anticonvulsant action of four conventional antiepileptic drugs (AEDs: clonazepam - CZP, ethosuximide - ETS, phenobarbital - PB and valproate - VPA) against pentetrazole (PTZ)-induced clonic seizures in mice. The results indicate that CAF at a dose of 92.4 mg/kg significantly reduced the threshold for PTZ-induced clonic seizures in mice from 69.5 to 51.7 mg/kg (p<0.05), being ineffective at lower doses of 69.3 and 46.2 mg/kg. Moreover, CAF at doses of and 92.4 mg/kg attenuated the protective action of ETS against PTZ induced seizures, by increasing its median effective dose (ED50) from 127.7 to 182.3 (p<0.05), and 198.3 mg/kg (p<0.01), respectively. In this case, no pharmacokinetic changes in total brain ETS concentrations after systemic ip administration of CAF (at 92.4 mg/kg) were observed, indicating a pharmacodynamic nature of interaction between ETS and CAF in the PTZ-test in mice. In contrast, CAF (at a dose of 92.4 mg/kg reducing the threshold for PTZ-induced seizures) combined with other AEDs (CZP, PB and VPA) did not affect their anticonvulsant action in the PTZ test in mice. Moreover, CAF (92.4 mg/kg) did not alter significantly total brain concentrations of the remaining AEDs (CZP, PB and VPA). The evaluation of potential acute adverse effects produced by AEDs in combination with CAF revealed that neither CAF (up to 92.4 mg/kg) administered alone nor combined with the studied drugs (at doses corresponding to their ED(50) values in the PTZ-test) affected motor performance of animals in the chimney test. In conclusion, the acute exposure to CAF may diminish the antiseizure protection offered by ETS in epileptic patients. Therefore, patients treated with ETS should avoid CAF. PMID- 17085858 TI - 7-Nitroindazole enhances dose-dependently the anticonvulsant activities of conventional antiepileptic drugs in the mouse maximal electroshock-induced seizure model. AB - 7-Nitroindazole (7NI, a nitric oxide synthase [NOS] inhibitor) administered intraperitoneally (ip), 30 min before the test, at doses ranging between 50-200 mg/kg, raised the threshold for electroconvulsions in mice. Linear regression analysis revealed that the doses increasing the threshold by 50% (TID50) and 100% (TID100) over the control value for 7NI were 115.2 and 173.4 mg/kg, respectively. Moreover, 7NI dose-dependently potentiated the anticonvulsant effects of four conventional antiepileptic drugs (AEDs: carbamazepine - CBZ, phenobarbital - PB, phenytoin - PHT, and valproate - VPA) in the mouse maximal electroshock-induced seizure (MES) model. 7NI at 50 mg/kg enhanced only the anticonvulsant effect of PB, whereas the drug at 75 and 100 mg/kg potentiated the antiseizure effects of PB, PHT and VPA, but not those of CBZ against MES-induced seizures. Only 7NI at 150 mg/kg enhanced considerably the antielectroshock action of all studied AEDs in the MES test. Pharmacokinetic evaluation of interactions between 7NI and the investigated AEDs revealed that 7NI (150 mg/kg; ip) did not alter total brain concentrations of conventional AEDs in mice. L-arginine (L-Arg - a natural precursor of NO; administered ip, 500 mg/kg, 60 min before electroconvulsions) did not reverse the activity of 7NI (150 mg/kg), but in contrast, it significantly potentiated the anticonvulsant action of conventional AEDs combined with 7NI (150 mg/kg). Pharmacokinetic increase in total brain AED concentrations was observed for the combinations of L-Arg (500 mg/kg) with 7NI (150 mg/kg) and PHT (by 32%; p<0.01) or VPA (by 22%; p<0.05). Neither total brain CBZ nor PB concentrations were altered following the co-administration of L-Arg (500 mg/kg) with 7NI (150 mg/kg). 7NI at doses of 100-200 mg/kg significantly impaired spontaneous ambulatory activity in mice subjected to the Y-maze task. The NOS inhibitor at doses of 50 and 75 mg/kg had no significant effect on locomotor activity of animals, although the number of arm entries within the 5 min of observational time was reduced. Finally, it can be concluded that the enhancement of anticonvulsive efficacy of CBZ, PB, PHT and VPA by 7NI alone or in combination with L-Arg in the MES test, deserves more attention and further neurochemical studies are required to elucidate the exact role of NO in the brain. PMID- 17085859 TI - Effect of intraarticular tramadol administration in the rat model of knee joint inflammation. AB - Local administration of exogenous opioids may cause effective analgesia without adverse symptoms from the central nervous system. Experiments show that peripheral antinociceptive effect of opioids is observed especially in inflammatory pain. The aim of the research was to estimate the effect of tramadol on nociceptive process at the level of peripheral nervous system, after its local administration in the model of knee joint inflammation. Tramadol was administered intraarticulary into the rat knee joint, before the inflammation as a preemptive analgesia and, for comparison, after the intraarticular injection of carrageenan. The research determined the influence of tramadol injection on pain threshold for thermal stimuli, development of inflammatory processes using the measurement of joint edema and motor function following the induction of knee joint inflammation in the rat. Functional assessment of knee joint with inflammation, in terms of rats' mobility and body position as well as joint loading and mobility were studied. The results of the experiments show that local administration of tramadol induces antinociceptive effect. The effect of tramadol, which elicits also a decrease in inflammatory edema, appears not only after its administration after carrageenan when inflammation was already present, but also in the case of its injection prior to carrageenan in the scheme of preemptive analgesia. The results of the described research show that not only morphine but also another opioid, tramadol, widely used in clinical practice, inhibits nociception, edema and functional impairment of the paw after its local application directly to the inflamed knee joint. PMID- 17085860 TI - Vinpocetine and piracetam exert antinociceptive effect in visceral pain model in mice. AB - The effect of vinpocetine or piracetam on thermal and visceral pain was studied in mice. In the hot plate test, vinpocetine (0.9 and 1.8 mg/kg), but not piracetam, produced a reduction in nociceptive response. Vinpocetine (0.45-1.8 mg/kg, ip) or piracetam (75-300 mg/kg, ip) caused dose-dependent inhibition of the abdominal constrictions evoked by ip injection of acetic acid. The effect of vinpocetine or piracetam was markedly potentiated by co-administration of propranolol, guanethidine, atropine, naloxone, yohimbine or prazosin. The marked potentiation of antinociception occurred upon a co-administration of vinpocetine and baclofen (5 or 10 mg/kg). In contrast, piracetam antagonized antinociception caused by the low (5 mg/kg), but not the high (10 mg/kg) dose of baclofen. The antinociception caused by vinpocetine was reduced by sulpiride; while that of piracetam was enhanced by haloperidol or sulpiride. Either vinpocetine or piracetam enhanced antinociception caused by imipramine. The antinociceptive effects of vinpocetine or piracetam were blocked by prior administration of theophylline. Low doses of either vinpocetine or piracetam reduced immobility time in the Porsolt's forced-swimming test. This study indicates that vinpocetine and piracetam possess visceral antinociceptive properties. This effect depends on activation of adenosine receptors. Piracetam in addition inhibits GABA-mediated antinociception. PMID- 17085861 TI - Role of adrenal gland hormones in antiinflammatory effect of calcium channel blockers. AB - Effects of amlodipine, lacidipine and nicardipine on acute phase of inflammation in the carregeenan model in intact rats were investigated in this study. In addition, the most effective dose of nicardipine that had the highest anti inflammatory impact was investigated in carregeenan test in adrenalectomized rats. The effective dose of nicardipine was tested in the chronic phase of inflammation in the model of cotton pellet granuloma, and its efficiency was compared with diclofenac sodium. Amlodipine at 5 and 10 mg/kg doses showed 61%, 80%, lacidipine 73%, 34% and nicardipine 38%, 87% inhibition of carrageenan induced inflammation, respectively. Nicardipine (10 mg/kg) and diclofenac sodium (25 mg/kg) showed 11.6% and 16.2% inhibition, respectively against carrageenan induced edema. Diclofenac at 10 mg/kg showed 43% inhibition of the inflammation. In cotton pellet test, antiproliferative effects of nicardipine (10 mg/kg) and diclofenac sodium (10 mg/kg) were evaluated as 60% and 39.5%, respectively. The obtained results showed that calcium channel blockers and diclofenac sodium significantly blocked acute and chronic phases of inflammation in intact rats, but in adrenalectomized rats calcium channel blockers and diclofenac sodium had no significant antiinflammatory effect. PMID- 17085862 TI - Imidazoline receptors in relaxation of acetylcholine-constricted isolated rat jejunum. AB - Since 20 years the concept of specific imidazoline receptors has remained controversial. The problem with imidazoline receptors is mostly due to their functional similarity to alpha-adrenoceptors. In this work, a pharmacodynamic model of isolated rat jejunum longitudinal muscle strips constricted with acetylcholine (Ach) was applied to separate functional properties of the two types of receptors. Relaxation of the preparation was measured as a function of concentration of 2-(benzofuranyl)-2-imidazoline (2-BFI), a specific imidazoline I2 receptor ligand, cirazoline, a potent I2 receptor ligand and alpha1 adrenoceptor agonist, phenylephrine, an agonist of alpha1-adrenoceptor, moxonidine, a ligand of I1 receptor, efaroxan, a ligand of I3 receptor and 5 bromo-6-(imidazoline-2-yl-amino)quinoxaline (UK14304), an agonist of alpha2 adrenoceptor. Next, the effects of a series of imidazoline-and/or alpha adrenoceptor-binding drugs (prazosin, yohimbine, RS79948, RX821002, idazoxan and efaroxan) on the relaxation of the Ach-constricted rat jejunum strips, induced by 2-BFI, cirazoline or phenylephrine, were studied. The obtained results demonstrate the involvement of the postsynaptic imidazoline receptors in rat jejunum motility. These receptors are of I2 subtype and are linked to alpha adrenoceptors of the predominantly alpha1 subtype. The alpha1 receptors dominate functionally over the I2 in the isolated rat jejnum. The proposed model might be useful in search for more specific new drugs. PMID- 17085863 TI - Influence of amlodipine and atenolol on lipopolysaccharide (LPS)-induced serum concentrations of TNF-alpha, IL-1, IL-6 in spontaneously hypertensive rats (SHR). AB - An increasing body of evidence suggests that cytokines may play a role in the pathogenesis of cardiovascular diseases. Immunopharmacological studies provide new information on immunomodulating activity of some drugs, including their effect on the level of pro-inflammatory cytokines. The aim of the present study was to find out whether amlodipine and atenolol, drugs applied in the treatment of arterial hypertension, can modulate lipopolysaccharide (LPS)-induced pro inflammatory cytokine level (TNF-alpha, IL-1, IL-6) in spontaneously hypertensive rats (SHR). The experiments were performed on 4 groups of animals as follows: WKY + MET(control Wistar-Kyoto normotensive rats), SHR + MET(control hypertensive rats), SHR + AML(hypertensive rats receiving amlodipine), SHR + AT (hypertensive rats receiving atenolol). Control rats received 1% solution of methylcellulose (1 ml/kg) by a gavage. Amlodipine and atenolol were administered by a gavage at doses of 15 mg/kg and 25 mg/kg, respectively. Arterial blood pressure was measured in conscious rats, using the tail-cuff method. Serum tumor necrosis factor alpha (TNF)-alpha, interleukin (IL)-1 and IL-6 concentrations were measured with enzyme-linked immunosorbent assay kits. Additionally, lipid levels were evaluated. The present data provide the evidence that amlodipine and atenolol act as immunomodulators of pro-inflammatory cytokines in SHR. Amlodipine decreased TNF-alpha, increased IL-6 and did not affect IL-1 level. Atenolol did not influence TNF-alpha and IL-1, but raised IL-6 in SHR. Additionally, amlodipine decreased total cholesterol level without changing HDL cholesterol level whereas atenolol did not influence lipid levels. The identification of additional immunomodulating properties of hypotensive drugs may be important for better understanding of their mechanisms of action. PMID- 17085864 TI - MDR1 (ABCB1) gene polymorphism C3435T is associated with P-glycoprotein activity in B-cell chronic lymphocytic leukemia. AB - Functional single nucleotide polymorphism (SNP) C3435T in exon 26 of the MDR1 ( ABCB1 ) gene encoding the xenobiotic transporter P-glycoprotein (P-gp, MDR1, ABCB1) may influence susceptibility to several diseases as well as clinical outcome of treatment with P-gp substrates. Exposure to environmental chemicals is thought to be involved in the pathogenesis of B-cell chronic lymphocytic leukemia (B-CLL) and P-gp-transported drugs are used in its treatment; however, little is known about the impact of the C3435T MDR1 SNP in B-CLL. In this study, 110 Caucasian B-CLL patients and 201 healthy controls were genotyped for the MDR1 C3435T SNP. Additionally, P-gp activity was assessed in malignant lymphocytes of 22 untreated B-CLL patients. We observed a higher frequency of carriers of at least one 3435T allele (3435CT and 3435TT genotypes) among B-CLL patients as compared to normal individuals (76% vs . 63%, p=0.027). The genotypes 3435CT and 3435TT were associated with B-CLL, (odds ratio=1.8, 95% confidence interval = 1.1 3.0). Moreover, P-gp activity in B-CLL cells depended on MDR1 genotype, with the highest P-gp activity in 3435CC homozygotes, intermediate in 3435CT heterozygotes and the lowest in 3435TT homozygotes (p=0.042). P-gp activity was also significantly lower in carriers of the T-allele (3435CT/TT genotype) as compared to the non-carriers (3435CC genotype), (p=0.029). Taken together, these data indicate that the MDR1 C3435T SNP may carry an increased risk of developing B CLL, possibly by virtue of decreased protection against P-gp-substrate carcinogens. The differences in P-gp activity in B-CLL tumor cells related to MDR1 genotype may have implications to the response to chemotherapy with P-gp transported anticancer agents. PMID- 17085865 TI - Extralipid effects of micronized fenofibrate in dyslipidemic patients. AB - The aim of this study was to evaluate the levels of lipid and extralipid parameters in patients with atherogenic dyslipidemia. We investigated the lipid lowering therapeutic efficacy of fenofibrate and its extralipid influence on oxidized low-density lipoprotein (oxLDL), C-reactive protein (CRP), Fibrinogen, factor VII and plasminogen activator type 1 (PAI-1) during 1-month treatment. Fourteen individuals with HLPIIb were treated with micronized fenofibrate (267 mg/d) for 1 month. The control group included twelve volunteers. Lipidograms were determined with enzymatic kits. ELISA method was used to measure oxLDL and PAI-1. Plasma CRP levels were measured spectrophotometrically. Fibrinogen and factor VII serum levels were evaluated with automatic coagulometer. After 1-month therapy with micronized fenofibrate, we observed a significant reduction of total cholesterol (TC) (277.2 to 217.8 mg/dl, p < 0.05), LDL (183.6 to 129.4 mg/dl, p < 0.05), trigliceryde (TG) (316.7 to 220.6 mg/dl, p < 0.05), oxLDL (68.7 +/- 5.5 to 39.7 +/- 3.7 U/l, p < 0.001) and increase in high-density lipoprotein (HDL) (35.1 to 41.9 mg/dl, p < 0.05). Fibrate treatment also decreased CRP(5.81 +/- 0.26 to 5.08 +/- 0.06 mg/l, p < 0.001), PAI-1 (120.4 +/- 9.7 to 84.7 +/- 5.9 ng/ml; p < 0.05), fibrinogen (3.65 +/- 0.17 to 3.44 +/- 0.16 g/l, ns) and factor VII (159.7% +/- 56.7 to 141% +/- 42.4, ns). The micronized fenofibrate at a daily dose of 267 mg demonstrated a highly beneficial effect on all lipid parameters and advantageous influence on inflammatory and thrombogenic plasma risk factors in patients with dyslipidemia HLPIIb. PMID- 17085866 TI - Comparison of the influence of halothane and isoflurane on airway transepithelial potential difference. AB - Bidirectional transport of Na+ and Cl- ions by the epithelium controls production and composition of airway surface liquid and airway transepithelial potential difference and in these ways supports mucociliary transport. Volatile anesthetics are able to inhibit epithelial ion transport processes when applied at high concentration and have been suggested to elicit depression of airway clearance and both these effects could be involved in postoperative pulmonary complications. The goal of these studies was to reveal possible influence of halothane and isoflurane at lower concentrations on electrogenic ion transport in airway epithelium. These studies were performed on the isolated rabbit tracheal wall mounted in the Ussing chamber. The reaction of the preparation to the gentle mechanical stimulation performed as a jet flux was examined without or in the presence of anesthetics at concentration equivalent to 0.5 minimal anesthetic concentration of volatile anesthetics in pulmonary alveoli (MAC), 1 MAC, 2 MAC, 5 MAC and 10 MAC. The volatile anesthetics at concentrations equivalent to 5 and 10 MAC affected airway transepithelial potential difference and influenced hyperpolarization or depolarization reactions which occurred after mechanical stimulation. The above effects were present when Na+ transport was inhibited by amiloride. The disturbed epithelial Cl- transport may be proposed as an explanation of the action of volatile anesthetics on electrophysiological parameters of the isolated tracheal wall although the influence of anesthetics on tachykinin secretion from C-fiber endings, which are present in the preparation, should also be taken into consideration. The long-lasting action (tens of minutes) of volatile anesthetics on the isolated tracheal wall should be also studied in the future as a model of airway reaction to prolonged volatile anesthesia. PMID- 17085867 TI - Immobility stress induces depression-like behavior in the forced swim test in mice: effect of magnesium and imipramine. AB - Previously, we demonstrated antidepressant-like effect of magnesium (Mg) in the forced swim test (FST). Moreover, the joint administration of Mg and imipramine (IMI) at ineffective doses per se, resulted in a potent reduction in the immobility time in this test. In the present study, we examined the effect of immobility stress (IS), and Mg and/or IMI administration on FST behavior. IS induced enhancement of immobility time, which was reversed by Mg or IMI at doses ineffective in non-stressed mice (10 mg/kg and 15 mg/kg, respectively). The joint administration of Mg and IMI was effective in both IS and non-stressed animals in FST. IS did not significantly alter locomotor activity, while IMI or Mg + IMI treatment in IS mice reduced this activity. We also measured serum and brain Mg, IMI and its metabolite desipramine (DMI) concentration in mice subjected to FST and injected with Mg + IMI, both restrained and non-restrained. In the present study we demonstrated a significant increase (by 68%) in the brain IMI and a slight, non-significant reduction in DMI concentration in IS + Mg + IMI + FST vs. Mg + IMI + FST groups, which might indicate the reduction in brain IMI metabolism. The IS-induced reduction in brain IMI metabolism did not participate in the activity in FST, since no differences in such activity were noticed between IS + Mg + IMI + FST and Mg + IMI + FST groups. The present data suggest that IS-induced increase in immobility time in FST is more sensitive for detection antidepressant-like activity. However, further studies are needed to examine the effect of other antidepressants in such an experimental paradigm. PMID- 17085868 TI - Medium supplementation with zinc enables detection of imipramine-induced adaptation in glycine/NMDA receptors labeled with [3H]L-689,560. AB - Antidepressant drugs after chronic administration induce adaptive changes in the NMDA receptor complex. Radioligand-receptorbinding studies using [3H]5,7 dichlorokynurenic acid demonstrated a "down-regulation" of the glycine site/NMDA receptor following chronic treatment with antidepressants and electroconvulsive shock. However, binding procedure using this radioligand is time consuming because it requires the use of centrifugation method in the separation process. The introduction of a new radioligand of glycine/NMDA receptor, [3H]L-689,560 enables the application of a rapid filtration method. In the present study we demonstrate that 2-week treatment with imipramine (15 mg/kg ip) did not evoke alterations in specific [3H]L-689,560 binding and in IC50 value of glycine in displacing [3H]L-689,560 binding in the mouse or rat cortex. However, longer, a 4 week treatment with imipramine induced a significant 71% increase in IC50 value in displacing [3H]L-689.560 binding in the mouse cortex. Moreover, the presence of zinc in the incubation media, dose-dependently enhances detection of imipramine-induced increase in IC50 value of glycine in displacing [3H]L-689,560 binding in the rat cortex. The present data indicate that: (1) [3H]L-689,560 may be a suitable ligand for assessing adaptation of the glycine/NMDA sites and (2) the presence of zinc enhances detection of imipramine-induced reduction of glycine affinity for glycine/NMDA receptors labeled with [3H]L-689,560 which further indicates a significance of zinc in the mechanism of antidepressant treatment. PMID- 17085869 TI - Antidepressant-like effects of baclofen and LY367385 in the forced swim test in rats. AB - Baclofen, an agonist of GABA(B) receptors and LY367385, an antagonist of mGluR(1a) receptors, given alone or jointly, reduced the immobility time in the forced swim test but only their separate administration enhanced motility in group of rats without hypoxia. Short-term hypoxia (2% O2, 98% N2, 4 min) did not change the activity of the rats in the forced swim test and it did robustly decrease the motility of these animals. LY367385 reduced the immobility time in the forced swim test but induced locomotion in rats subjected to hypoxia. The obtained results indicated that baclofen and LY367385 given alone or jointly induce an antidepressant-like effect in the forced swim test but only LY367385 possesses such activity in rats that had undergone hypoxia. Both tested ligands are involved in the motility of rats, however, LY367385 influences hypolocomotion hypoxia-induced. PMID- 17085870 TI - Mebeverine influences sodium ion transport in the distal colon. AB - The study was performed to check if the well-known intestinal spasmolytic effect of mebeverine is paralleled by any changes in intestinal transepithelial currents. The transepithelial potential difference related to ionic currents of the isolated rabbit distal colon wall was measured by means of Ussing's technique under control conditions and after gentle mechanical stimulation of intestinal epithelial surface by a flux from peristaltic pump and with and without of mebeverine in stimulation fluid. The transient hyperpolarization after mechanical stimulation was diminished after addition of mebeverine to the stimulation fluid when chloride transport was inhibited by bumetanide (BUME) but in the presence of amiloride (AMI), a sodium ion transport inhibitor, the drug did not influence the reaction. It was inferred that mebeverine was able to modulate transepithelial sodium ion transport and in this way to modify interaction between colonic wall and its contents during intestinal passage. PMID- 17085871 TI - Cerebral oligemic hypoxia and MK-801 treatment: effect on alternation behavior in mice. AB - It is known that glutamatergic system is one of neurotransmitter systems affected by a transiently reduced oxygen supply. The aim of the present study was to examine the effects of MK-801, a noncompetitive NMDA receptor antagonist, on spontaneous alternation in mice exposed to cerebral oligemic hypoxia. Spontaneous alternation behavior and locomotor activity were evaluated using the Y-maze task. Transient cerebral oligemia was induced by bilateral clamping of carotid arteries (BCCA) for 30 min under pentobarbital anesthesia. MK-801 was injected 48 h after BCCA or sham surgery, 30 min before the test session. Treatment with MK-801 (0.1 mg/kg ip) impaired spontaneous alternation both in sham-operated and BCCA mice. MK-801 (0.1 mg/kg ip) significantly enhanced the locomotion of mice. The effects of MK-801 were not exacerbated by BCCA. These results show that cerebral oligemic hypoxia induced by BCCA does not change alternation behavior of mice treated with MK-801. PMID- 17085872 TI - Social class inequalities in self-rated health and their gender and age group differences in Japan. AB - BACKGROUND: Few studies have examined social inequalities in self-rated health in Japan, and the issue of gender differences related to social inequalities in self rated health remains inconclusive. METHODS: The data derived from interviews with 2987 randomly selected Japanese adults in four prefectures in Japan who completed the cross-national World Mental Health survey from 2002 through 2005. We calculated odds ratios (ORs) of having poor self-rated physical and mental health by two social class indicators independently with multivariate logistic regression models, adjusted for age, gender, marital status, and area. Stratified analyses by gender and age group were also conducted. RESULTS: The adjusted ORs of the lowest educational attainment category having poor self-rated physical and mental health were 1.42 (95% confidence interval [CI]: 1.15-1.76) and 1.37 (95% CI: 1.10-1.70), respectively. Among females, educational attainment had significant linear associations with self-rated physical and mental health. Adjusted household income was also significantly associated with self-rated physical health among female respondents. No associations were found among males. While educational attainment was associated with self-rated health among the young age group, adjusted household income was associated with self-rated physical health in the middle and old age group. CONCLUSION: These results indicated social inequalities in self-rated health and prominent social inequalities in self-rated health among females in Japan. Social inequalities in self-rated health seemed to exist across age groups. However, the mechanism of social inequalities in self-rated health could be different depending on the age group. PMID- 17085873 TI - hOGG1 Ser326Cys polymorphism and risk of hepatocellular carcinoma among Japanese. AB - BACKGROUND: The Ser326Cys polymorphism in human oxoguanine glycosylase 1 (hOGG1), which is involved in the repair of 8-hydroxy-2-deoxyguanine in oxidatively damaged DNA, has been associated with susceptibility to certain cancers, but has not been examined in causation of hepatocellular carcinoma (HCC). METHODS: We conducted a case-control study to investigate whether this polymorphism was related to HCC risk with any interaction with alcohol consumption and cigarette smoking. Genotyping was performed by a polymerase chain reaction with confronting two-pair primers among 209 newly diagnosed HCC cases, 275 hospital controls, and 381 patients with chronic liver disease (CLD) without HCC. RESULTS: Overall, the hOGG1 genotype was not significantly associated with HCC; adjusted odds ratios (and 95% confidence intervals) for the Ser/Cys and Cys/Cys genotypes compared with the Ser/Ser genotype were 0.79 (0.35-1.79) and 0.48 (0.18-1.27) against hospital controls, and 1.51 (0.96-3.37) and 0.86 (0.50-1.47) against CLD patients. We could not detect any significant gene-alcohol interaction (p = 0.95 or 0.16) or gene-smoking interaction (p = 0.70 or 0.69). CONCLUSIONS: These results suggest that the hOGG1 Ser326Cys polymorphism may not play a major role as an independent factor in hepatocarcinogenesis. PMID- 17085874 TI - Increase of colon and rectal cancer incidence rates in Japan: trends in incidence rates in Miyagi Prefecture, 1959-1997. AB - BACKGROUND: During recent decades, colorectal cancer incidence rates have been rapidly increasing in Japan. To investigate trends in colorectal cancer incidence rates, we analyzed incidence data during 39 years between 1959 and 1997 in Miyagi Prefecture, Japan. METHODS: Using age-period-cohort models, we evaluated the effects of time period and cohort on colon and rectal cancer incidence. Model fitting was based on eleven 5-year age groups (30-34 to 80-84), eight 5-year time periods, and 18 overlapping birth cohorts of 10 years each. RESULTS: The analysis found a significant (p=0.04) and upward period effect on female colon cancer incidence, and a significant (p<0.01) and upward cohort effect on male colon cancer incidence. An upward period effect was also observed for male colon cancer incidence without significance. For rectal cancer incidence, a significant cohort effect was found among both males and females. CONCLUSIONS: In light of known risk factors of colorectal cancer, the effects of period and cohort might be related to the change in the prevalence of risk factors such as high intake of meat and animal fat, and obesity. The improved diagnostic procedures including the spread of cancer screening might be responsible for the period effect. Although the significant cohort effects may give a caution for a continuous increase of colorectal cancer incidence, the future trend may be influenced by the period-related factors. Successive monitoring of cancer incidence and prevalence of risk factors is required. PMID- 17085875 TI - Dietary supplement use by community-living population in Japan: data from the National Institute for Longevity Sciences Longitudinal Study of Aging (NILS-LSA). AB - BACKGROUND: There are few studies about dietary supplement use and nutrient intake from these products in Japan. The purpose of this study was to clarify (1) the prevalence of dietary supplement use, (2) the characteristics of dietary supplement users, (3) nutrient intake from dietary supplements, and (4) the existence of dietary supplement users who took excessive nutrients from these products. METHODS: To collect the information on dietary supplement use in the previous year and nutrient intake from these products, we conducted a self administered dietary supplement frequency questionnaire. The subjects were 2,259 people aged 40-82 years. Dietary supplements were grouped into 8 major categories. A dietary supplement database was developed to estimate nutrient intake from these products. Excess users were defined as people who consumed more nutrient than the tolerable upper intake level of the Dietary Reference Intakes for Japanese. RESULTS: In the previous year, 55 % of males and 61 % of females consumed dietary supplements. Dietary supplement use was especially prevalent in females, subjects who felt unhealthy, and subjects who were more careful of maintaining an appropriate weight, though the association was affected by the frequency of dietary supplement use. The most common dietary supplements were drink type in males and vitamins in females. Some nutrient values obtained from dietary supplements were higher than those from food. Excess users were found for intake of vitamin A, B(6), K, niacin, iron, and magnesium. CONCLUSIONS: It is important to clarify dietary supplement use and to estimate nutrient intake from these products. PMID- 17085876 TI - Frequency of going outdoors as a good predictors for incident disability of physical function as well as disability recovery in community-dwelling older adults in rural Japan. AB - BACKGROUND: The clinico-epidemiologic relevance of the reduction in the frequency of going outdoors in older adults has not been well characterized. This study examined whether the frequency of going outdoors has predictive values for incident physical disability and recovery among community-dwelling elderly. METHODS: One thousand, two hundred and sixty-seven persons aged 65+ years who lived in a rural community in Niigata, Japan, and participated in the baseline survey were assessed again 2 years later in terms of mobility, and instrumental and basic activities of daily living (IADL and BADL). We compared the incident disability and recovery at follow-up among three subgroups classified by the baseline frequency of going outdoors: once a day or more often, once per 2-3 days, and once a week or less often. Multivariate analyses tested associations between the frequency of going outdoors and functional transition, independent of potential confounders. RESULTS: A lower frequency of going outdoors at baseline was associated with a greater incident disability, and a lower recovery at the two-year follow-up. Even after adjustment, the effects of going outdoors remained significant. Adjusted risks of incident mobility and IADL disabilities were significantly higher (odds ratio[OR]=4.02, 95% confidence interval [CI]: 1.77 9.14 and OR=2.65, 95% CI: 1.06-6.58), respectively, and recovery from mobility disability was significantly lower (OR=0.29, 95% CI: 0.08-0.99) for "once a week or less often" subgroup compared with "once a day or more often" subgroup. CONCLUSION: The frequency of going outdoors is a good predictor for incident physical disability and recovery among community-living elderly. Public health nurses and clinicians should pay more attention how often their senior clients usually go outdoors. PMID- 17085877 TI - Ethanol exposure decreases cell proliferation and increases apoptosis in rat testes. AB - Ethanol exposure is known to suppress male reproductive activity in laboratory animals and humans. The present study was designed to evaluate whether chronic ethanol exposure decreases proliferative activity or increases apoptosis in the testes. Ethanol (1.5 g/kg or 3 g/kg i.p., 15% v/v in saline) was administrated to adult male rats for 10 days. Proliferating cell nuclear antigen (PCNA) was used as a proliferative marker. Western blot analysis showed that ethanol administration significantly reduced the level of PCNA. Also, immunoreactivity of PCNA-positive cells in the spermatogonia and primary spermatocytes were decreased by ethanol exposure. However, the number of TUNEL-positive cells was significantly increased in the testicular germ cells of ethanol-treated rats. Moreover, ethanol administration significantly increased the level of activated caspase-3 in testes. In conclusion, our findings suggest that ethanol may partly contribute to the suppression of male reproductive activity through a reduction of cell proliferation and an enhancement of cell death in rat testes. PMID- 17085878 TI - 17beta-estradiol pretreatment prevents the global ischemic injury-induced decrease of Akt activation and bad phosphorylation in gerbils. AB - Estradiol acts as a neuroprotective factor against ischemic brain injury. This study investigated whether estradiol modulates neuroprotective mechanism through the activation of Akt and its downstream target such as Bad in global ischemic injury. Adult female gerbils were ovariectomized and treated with estradiol prior to ischemic injury. Transient cerebral ischemia was accomplished by bilateral clipping of the common carotid artery for 5 min. Brains were collected on 1, 3, 5 day after injury. In hippocampal CA1 region of non-treated gerbils, most of neuronal cells exhibited pyknotic nuclei and showed the positive reaction of TUNEL staining on 5 day after injury. However, estradiol significantly reduced the neuronal cell death. Potential activation was measured by phosphorylation of Akt at Ser473 and Bad at Ser136 using western blot analysis. The levels of pAkt and pBad were significantly decreased in non-treated gerbils on 1-5 day after injury. However, estradiol prevents the global ischemic injury-induced decrease of pAkt and pBad. Our findings suggest that estradiol prevents cell death due to global ischemic injury and that Akt activation and Bad phosphorylation by estradiol mediated these protective effects. PMID- 17085879 TI - Differentiation of epithelial cells to M cells in response to bacterial colonization on the follicle-associated epithelium of Peyer's patch in rat small intestine. AB - To clarify the relationship between M cells and intestinal microflora, histoplanimetrical investigation into the bacterial colonization and the differentiation to M cells was carried out in rat Peyer's patch under physiological conditions. The follicle-associated epithelium (FAE), except for the narrow area of apical region, was closely covered with both neighboring intestinal villi and a thick mucous layer, the latter of which also filled the intervillous spaces as well as the space between the FAE and the neighboring intestinal villi. Indigenous bacteria adhered almost constantly to the narrow areas of apical regions of both intestinal villi and the FAE. Bacterial colonies were occasionally located on the basal to middle region of FAE, where M cells also appeared, forming large pockets. When bacterial colonies were located on the basal to middle region of FAE, bacteria with the same morphological characteristics also proliferated in the intervillous spaces neighboring the Peyer's patch. In cases with no bacterial colonies on the basal to middle region of FAE, however, M cells were rare in the FAE. Histoplanimetrical analysis showed the similar distribution pattern of bacterial colonies on the FAE and M cells in the FAE. M cells ultrastructurally engulfed indigenous bacteria, which were then transported to the pockets. These results suggest that indigenous bacterial colonization on the FAE stimulates the differentiation of M cells in the FAE under physiological conditions. The uptake of bacteria by M cells might contribute the regulation of the development of indigenous bacterial colonies in the small intestine. PMID- 17085880 TI - Pathogenicity and immunogenicity of equine herpesvirus type 1 mutants defective in either gI or gE gene in murine and hamster models. AB - To develop a live vaccine for equine herpesvirus type 1 (EHV-1), two EHV-1 mutants containing no heterogeneous DNA, DeltagI and DeltagE, were constructed with deletions in the open reading frame of either glycoprotein I (gI) or E (gE), respectively. In equine cell culture, deletion mutants formed smaller plaques than the parental and revertant viruses, but the one-step growth patterns of the deletion mutants and the parental strain were approximately the same. These results suggest that both gI and gE contribute to the ability of EHV-1 to spread directly from cell-to-cell, but that these glycoproteins are not required for viral growth in vitro. Mice and hamsters inoculated intranasally with these mutants showed no clinical signs, and continued to gain weight, whereas those inoculated with the parental virus exhibited a reduction in mean body weight. Furthermore, nervous manifestations were observed in hamsters inoculated with the parental virus. These results suggest that gI and gE have an important role in EHV-1 virulence including neurovirulence in experimental animal models. On the other hand, serum neutralizing antibodies were detected in mice immunized with DeltagI or DeltagE at two weeks after inoculation. Following challenge with the parental virus, DeltagI- or DeltagE-immunized mice were able to clear parental virus from their lungs faster than mock-immunized mice. These results suggest that the EHV-1 mutants defective in gI and in gE are attenuated but have ability to elicit immune responses in inoculated mice that contribute to virus clearance. PMID- 17085881 TI - Effects of high-K+, Na+-deficient solution on contractility of the smooth muscles of the bovine trachea. AB - A high-K+, Na+-deficient (I-154 K+) solution induced contraction followed by gradual relaxation of the smooth muscles of the bovine trachea, while hyperosmotic addition of 65 mM KCl induced a large sustained contraction. Exposure of the muscle to the I-154 K+ solution induced an increase in the ratio of cellular water content and a sustained increase in oxidized flavoprotein fluorescence or reduced pyridine nucleotide fluorescence. The I-154 K+ solution also induced a sustained increase in [Ca2+]i level. Decreases in developed tension and increases in cellular water content were both prevented by the addition of sucrose or NaCl but not pyruvate. Substitution of KI for KCl in the I 154 K+ solution produced a greater inhibition of contraction, while substitution with K-propionate produced no inhibition of contraction. Moreover, decreases in developed tension and increases in cellular water content were both prevented by addition of 100 microM 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS), but not 10 microM bumetanide or 1 mM acetazolamide. In conclusion, I-154 K+ solution induced-relaxation in the bovine trachea may be due to swelling of smooth muscle cells and the mechanism of swelling is probably involved in DIDS sensitive anion movement. PMID- 17085882 TI - Establishment and characterization of a cell line, MCO-Y4, derived from canine mammary gland osteosarcoma. AB - A cell line, MCO-Y4, was established from a mammary gland osteosarcoma of a 16 year-old female mongrel dog. Histopathologically the tumor was composed of osteoblastic cells with an osteoid meshwork and chondroid matrix. The mean doubling time of the cells at the 93rd passage was 32.39+/-4.66 hr. Immunohistochemically, the osteoblastic and chondroblastic cells were positive for bone morphogenetic protein (BMP)-2/4 and BMP receptor (BMPR) II. The cultured cells were spindle in shape during the growth and the confluent phases. No tumor matrix was detected in the culture dish by alcian blue staining or von-Kossa silver impregnation. MCO-Y4 cells on the chamber slides showed intense immunoreactivity for BMP-2/4 and BMPR II. Noggin, an antagonist for BMP-2/4, showed the growth inhibition on MCO-Y4 cells. In addition, fibronectin might be potential for stimulating growth of MCO-Y4 cells. When transplanted into severe combined immunodeficiency mice, the cells formed tumors consisting of solid proliferation of osteoblastic and fibroblastic cells with woven-bone trabeculae. These tumor cells were intensely positive for BMP-2/4 and BMPR II. Our results suggested that the cell line might be useful for studying the role of BMPs in canine osteosarcoma and the mechanism of ossification. PMID- 17085883 TI - Effects of liquid nitrogen vapor sensitization conditions on the quality of frozen-thawed dog spermatozoa. AB - The freezing conditions for preparation of frozen canine semen by the plunging method were investigated with regard to the period of sensitization in liquid nitrogen (LN2) vapor and the height from LN2, and the semen qualities after thawing were compared with those of canine semen prepared by the simple freezer method previously reported by us. In the plunging method, 9 semen straws were prepared under the same conditions, horizontally kept at 5, 7, and 10 cm above the LN2 surface in a styrene foam box for 5, 10, and 15 min, and then plunged into LN2. The semen qualities immediately after thawing were high in the 7 cm/10 min (cooling rate: -4 to -22 degrees C/min) and 10 cm/15 min groups (cooling rate: -6 to -10 degrees C/min). On comparison of frozen semen prepared by the plunging method (7 cm/10 min) with frozen semen prepared by the simple freezer method, sperm motility and viability were significantly higher for the frozen semen prepared by the plunging method. The cooling rate in freezing was higher for the simple freezer method (cooling rate: -6 to -50.9 degrees C/min) than the plunging method. Based on these findings, horizontal placement of canine semen straws above LN2 to reduce the temperature at a slow cooling rate of about -10 degrees C/min, followed by plunging into LN2 after sensitization for 10-15 min, provides good semen qualities after thawing. PMID- 17085884 TI - Peripheral mononuclear cell response in Japanese black calves after oral administration of IFN-alpha. AB - To investigate the effects of oral administration of an interferon (IFN)-alpha drug on the immune reaction of healthy Japanese Black (JB) calves, peripheral leukocyte populations and their ability to produce cytokine mRNA were analyzed after oral administration of IFN-alpha. Fourteen calves fed in one herd were divided into two groups; seven calves were orally administered 0.1 g/day of IFN alpha from the day of birth to day 5 on each day (group 1, N=7), and the other seven calves were used as the control (group 2, N=7). Blood samples were collected from the jugular veins of all calves before administration and in weeks 1, 2, 4, 8, and 16 after birth. The number of MHC class II+CD14+ monocytes in the leukocytes population of group 1 increased gradually after birth, and significantly higher numbers were detected in week 4 compared with group 2. MHC class II-CD14+ monocytes in group 1 peaked in week 1, and a significant increase was detected compared with group 2. The level of IL-12 in the cytokine mRNA of group 1 increased gradually between weeks 1 and 2, and a significantly higher level of IL-12 was found compared with group 2. These results suggest that oral administration of IFN-alpha induces activation of the monocyte functions in JB calves. PMID- 17085885 TI - Effect of echo-guided high-intensity focused ultrasound ablation on localized experimental tumors. AB - We fabricated a prototype 3.25-MHz split-focus therapeutic transducer combined with a small 6.5-MHz imaging ultrasonic probe for transrectal treatment of prostate cancer and evaluated the feasibility of using split-focus high-intensity focused ultrasound (HIFU) to ablate localized tumor tissue without injuring the surrounding organs. We therefore established a localized tumor model by inoculating VX2 tumor into rabbit livers. The localized VX2 tumors of nine rabbits were transdermally treated with split-focus ablation at a peak intensity in water of 6 kW/cm2 for 4 s (6 shots) under the guidance of ultrasonic B-mode imaging. Necropsy a day after treatment found the surface of the livers and gastrointestinal tracts to be grossly normal. The VX2 tumors were completely coagulated and were surrounded by ablated liver tissue. The six shots of split focus HIFU destroyed the VX2 tumors without injuring the liver surfaces or the surrounding organs. These results suggest that split-focus HIFU ablation could be an effective treatment of localized tumors. PMID- 17085886 TI - Effect of melatonin injected into the third ventricle on growth hormone secretion in Holstein steers. AB - The effects of melatonin (MEL) injection into the third ventricle (3V) on growth hormone (GH) secretion were investigated in conscious Holstein steers. A stainless steel cannula was stereotaxically implanted in the 3V based on the ventriculogram. In Exp. 1, three doses of MEL (100, 300 or 600 microg) were injected into the 3V through the cannula and the GH concentration after the injection was determined. In Exp. 2, intracerebroventricular (icv) and intravenous (iv) injections of MEL (100 microg) and GH-releasing hormone (GHRH; 0.25 microg/kg body weight), respectively, were performed simultaneously to examine the effect of MEL on GHRH-induced GH release. The icv injection of MEL significantly stimulated GH release at 100 microg. The increase in GH concentrations by 100 microg of MEL was persistent. Intravenous injection of GHRH dramatically increased GH release. The injection of MEL did not alter GHRH induced GH release. These results suggest that MEL stimulates GH secretion possibly through the hypothalamus in cattle. PMID- 17085887 TI - Interaction between glycogen body cell and neuron: examination in co-culture system. AB - The glycogen body (GB) is in the dorsal area of the lumbosacral spinal cord in birds and is composed of uniform cells characterized by high glycogen storage. The glycogen of GB cells remains unchanged in vivo by the effects of a variety of hormones such as insulin, glucagon, adrenocorticotropic hormone and by physiological conditions such as starvation. In order to investigate the latent functionability of GB cells, we observed morphological changes of glycogen body cells in a co-culture system with cerebellar neurons by light and transmission electron microscopy. Cultured GB cells were labeled with 1,1'-dioctadecyl 3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI). The cultured neurons derived from cerebellum were co-cultured with the labeled GB cells. Under the co culture with neurons, 2 types of GB cells were detected. One was conventional with numerous glycogen deposits in the cytoplasm and tended to make clusters. The other type of GB cells singly extended the processes attaching to the neuronal body and axons. In the axons in contact with GB cell processes, small vesicles appearing as synaptic vesicles were observed. These observations suggested that some GB cells can differentiate to an average astrocyte. The GB cells were assumed to involve the synapse formation or maturing as astrocytes in the CNS. PMID- 17085888 TI - The specific antibody to Brachyspira aalborgi in serum obtained from a patient with intestinal spirochetosis. AB - Serum obtained from a patient histopathologically diagnosed as intestinal spirochetosis was investigated serodiagnostically by agglutination test. B. aalborgi which is a human intestinal spirochete reacted strongly with the human serum, while B. pilosicoli which has potential pathogenicity to humans reacted with the serum, but as strongly and its titer was different than the other three species. On the other hand, intestinal spirochetes (Matsumoto isolates) were isolated from the biopsy samples of the patient. The morphological, biochemical, and genetic characteristics of the isolates were very similar to those of B. aalborgi. Furthermore, the protein profiles of the Matsumoto isolates were also similar to those of B. aalborgi but were different than those of B. pilosicoli and B. hyodysenteriae. The reaction profiles of the Matsumoto isolates in immunoblotting were relatively similar to those of B. aalborgi except for a 74 kDa band but were different from those of B. pilosicoli and B. hyodysenteriae. Therefore, we identified the Matsumoto isolates as B. aalborgi and diagnosed the patient with a B. aalborgi infection. PMID- 17085889 TI - Annual changes in testicular size and serum and fecal testosterone concentrations in male bharals, Pseudois nayaur. AB - The serum and fecal testosterone (T) concentrations and testicular sizes of two male bharals (Pseudois nayaur) were determined for approximately one year. The profiles of the fecal T concentrations showed a similar tendency as the profiles of serum T concentrations, and there was a significant correlation between serum and fecal T concentrations (r=0.72). T concentrations rose drastically in October and decreased gradually until January. The maximum testicular size was observed between November and January. Semen collected between December and January was excellent in quality and comparable to domestic sheep and goats. The active periods of the testes were synchronized with the early breeding season of females. PMID- 17085890 TI - Geographical distribution of vampire bat-related cattle rabies in Brazil. AB - Seventy-seven rabies virus (RV) isolates originating from Brazilian cattle were genetically characterized. Partial nucleoprotein gene sequences of these isolates were phylogenetically and geographically analyzed. Cattle isolates, which clustered with the vampire bat-related RV group, were further subdivided into nine genetic subgroups. These subgroups were distributed widely in lowland regions, with some subgroups separated from each other by mountain ranges. In addition, separation of the groups in mountainous regions was correlated with altitude. These results indicate that cattle rabies is derived from several regionally-defined variants, which suggests that its geographical distribution is related to that of the vampire bat population. PMID- 17085891 TI - Characteristics of frozen-thawed spermatozoa cryopreserved with different concentrations of glycerol in captive Japanese black bears (Ursus thibetanus japonicus). AB - Seven mature Japanese black bears were used as semen donors, and a total of 7 semen samples collected from the animals by the electroejaculation method were cryopreserved in liquid nitrogen. Egg yolk-TRIS-citrate-glucose extender was used, and the effects of different final concentrations of glycerol, at 4-12% (v/v), on frozen-thawed spermatozoa were examined. No significant difference was observed in percent motility or percent abnormal morphology of frozen-thawed spermatozoa among the different glycerol concentrations. Percent viability and percent intact acrosomes of spermatozoa cryopreserved with 4 and 6% glycerol were significantly higher than those with 10 and 12% glycerol. These results suggest that a suitable glycerol concentration for freezing Japanese black bear semen within the range tested would be 4-6%. PMID- 17085892 TI - Genetic characterization of GRA6 genes from Toxoplasma gondii from pigs in Okinawa, Japan. AB - Toxoplasma gondii from pigs in Okinawa Prefecture was characterized by nested PCR restriction fragment length polymorphism (RFLP) and DNA sequence analysis of the dense granule antigen GRA6 gene. By nested PCR, parasite DNA was detected in 33 out of 91 lymph node samples with lesions similar to those found in toxoplasmosis samples that had been collected from pigs at an abattoir. RFLP analysis with MseI was successfully conducted in 29 of 33 PCR-positive samples to group the isolates into one of the three genotypes of T. gondii. Genotyping of the 29 studied samples rendered the following results: 13 of type I (44.8%), 14 of type II (48.3%), and 2 of type III (6.9%). The GRA6 genes of 12 Okinawa isolates were cloned and sequenced. Nine new nucleotide sequences were found, and nucleotide substitutions specific for the Okinawa isolates were found at 13 positions. Phylogenetic analysis indicated that all GRA6 sequences were divided into one of the 3 main groups, and Okinawa isolates of GRA6 genotypes II and III seemed to be closely related to the Beverley strain and the NED strain, respectively. The results from this study may provide basic and useful information for the analysis of the molecular epidemiology of T. gondii infection within Japan. PMID- 17085893 TI - Characterization of macrolide-resistant Campylobacter coli isolates from food producing animals on farms across Japan during 2004. AB - We investigated the susceptibilities against 7 antimicrobial agents in Campylobacter jejuni and C. coli isolates from food-producing animals in 2004. In comparison with the results of past surveillance, no significant difference was observed in resistance rates against all of the antimicrobials tested in Campylobacter isolates. However, slight increase of erythromycin (EM) resistance was found in C. coli isolates from pigs. We examined the mutation of the 23S rRNA gene and their susceptibilities against azithromycin, tylosin, and lincomycin in 44 EM-resistant isolates and 28 susceptible isolates of porcine origin. All the EM-resistant isolates contained A2075G in the 23S rRNA gene and showed cross resistance to azithromycin, tylosin, and lyncomycin. PMID- 17085894 TI - Effects of feeding calves genetically modified corn bt11: a clinico-biochemical study. AB - Genetically modified corn Bt11 is insect-resistant and expresses Cry1Ab toxin, an insecticidal protein, in kernels. Although Bt11 corn is considered safe based on animal performance, there are no reports available on the clinico-biochemical effects of feeding it to cattle. In this study, we evaluated the effects of feeding Bt11 to calves, using blood and ruminal clinico-biochemical parameters. Our three-month-long feeding experiment demonstrated that calves (n=6), fed with a ration containing 43.3% of Bt11 corn kernels as dry matter, did not develop any discernible clinical, hematological, biochemical, or ruminal abnormalities as compared with control calves (n=6) fed non-Bt11 corn. The results suggest that the transgenic Bt11 has no negative clinico-biochemical effects on calves. PMID- 17085895 TI - Systemic candidiasis in a dog, developing spondylitis. AB - A 4-year-old male Shiba dog initially presented with pain of an undetermined origin and hypersensitivity to touch. Seven days later, the dog developed ataxia, hind-leg weakness and knuckling. The dog died on 20 days after presentation. Postmortem examination revealed a mass in the body of thoracic vertebra. Histopathologically, the mass consisted of granulomatous inflammation, including fungal organisms that were immunohistochemically positive for Candida albicans. Similar granulomatous lesions were observed in the systemic lymph nodes, kidneys, pancreas, spleen, prostate gland, thyroid glands and heart. This case was diagnosed as systemic candidiasis with spondylitis. PMID- 17085896 TI - Identification of mutation in the growth differentiation factor 5 (Gdf5) gene in NC-brp/brp mice. AB - A brachypodism (brp) mutation arose spontaneously in the inbred NC mouse strain, producing a phenotype similar to that caused by bp mutation; therefore, it is strongly suggested that brp and bp are allelic. A series of bp mutations are due to defects in the growth differentiation factor 5 (Gdf5) gene. Nucleotide sequence analysis on the Gdf5 gene in NC-brp/brp mice revealed that an irregular insertion of a unit ;GGCAGCC' in exon 2 caused a frame shift leading to a premature stop codon. In addition to the known physiologic roles of brp, I found that brp significantly reduced the litter size. The brp is a novel mutant allele at the Gdf5 gene locus; I would like to name this allele Gdf5(brp). PMID- 17085897 TI - Effects of addition of sodium lauryl sulfate on frozen-thawed canine spermatozoa. AB - The addition of Orvus ES paste (OEP) to extender may be essential for preparing frozen dog semen. The major ingredient of OEP is sodium lauryl sulfate (SLS). In this study, we compared the effect of SLS on frozen dog semen with that of OEP. There were no significant differences between the 2-mg/ml SLS group and OEP group concerning sperm motility, viability and the percentage of viable sperm with intact acrosomes after freeze-thawing. These results suggest that the effectiveness of frozen dog semen extender containing 2 mg/ml of SLS is similar effective to that demonstrated for OEP. PMID- 17085898 TI - Canine serum thermostable alkaline phosphatase isoenzyme from a dog with hepatocellular carcinoma. AB - A dog histopathologically diagnosed with hepatocellular carcinoma (HCC) showed very high serum alkaline phosphatase (ALP) activity. A supernatant of ascitic fluid and tumor tissue extracted from the dog also showed much higher ALP activity than normal. ALP isoenzyme analysis of samples was performed using polyacrylamide gel disk electrophoresis, and a wide, broad abnormal band was observed. By various treatments, the abnormal band showed thermostability, which is a characteristic of tumor-associated ALP that has only been reported in humans. The thermostable ALP isoenzyme was not found in sera from 39 dogs with several types of tumor that originated from the liver, except for HCC, nor was it found in 10 dogs with hepatic diseases that did not include hepatic tumors. The thermostable ALP isoenzyme seemed to be associated with canine HCC. PMID- 17085899 TI - Incidence of ovulation without coital stimuli in captive Japanese black bears (Ursus thibetanus japonicus) based on serum progesterone profiles. AB - The purpose of this study was to ascertain whether or not Japanese black bears (Ursus thibetanus japonicus) are induced ovulators. The progesterone levels of female bears kept with a male and allowed to mate (n=2) and female bears allowed contact with a male through bars but not allowed to mate (n=6) during the mating season increased significantly in late October. Based on this result, the female bears were considered to have ovulated. Females isolated from males (n=3) were ovarioectomized after the mating season, and no corpora lutea were observed, indicating they had not ovulated. These findings suggest that Japanese black bears may be induced ovulators that ovulate with stimuli from males and without coitus at a high rate. PMID- 17085900 TI - A serological survey of Brucella spp. in free-ranging wild boar (Sus scrofa leucomystax) in Shikoku, Japan. AB - Brucella, a causative agent of brucellosis and facultative intracellular pathogen, has been isolated recently from a variety of wild mammals. In this study, serum samples from 115 Japanese wild boar (Sus Scrofa leucomystax) killed by hunters in the 4 Prefectures of Shikoku, Japan were tested for antibodies to Brucella spp. by means of the tube agglutination test (TAT) and enzyme-linked immunosorbent assay (ELISA) using antigens extracted with n-lauroylsarcosine. In 9 of the 115 samples (7.8%) antibodies to Brucella spp. were detected by TAT and ELISA. These results suggest that wild boar in Shikoku may be exposed to Brucella spp. or other cross-reactive pathogen infection. PMID- 17085901 TI - [Janus-faced function of tumor suppressor genes as a new paradigm of cancer research--link between metabolic syndrome and carcinogenesis]. PMID- 17085902 TI - [Strategy for acid suppressant-resistant NERD]. PMID- 17085903 TI - [The treatment for reflux esophagitis which is refractory to proton pump inhibitor therapy]. PMID- 17085904 TI - [How to handle Barrett's esophagus]. PMID- 17085905 TI - [A case of esophageal arterial bleeding in which transcatheter arterial embolization was useful for hemostasis]. AB - We report a case of hemostatic treatment by transcatheter arterial embolization (TAE) of the left gastric artery and a branch of the left bronchial artery for oozing hemorrhage in the lower esophagus, because of incomplete endoscopic hemostasis. A 52-year-old woman with systemic lupus erythematosus was previously admitted three times to our hospital for endoscopic treatment of hemorrhage in the lower esophageal lesions. The procedure of TAE for esophageal hemorrhage seems to be a useful treatment for cases of incomplete endoscopic hemostasis. PMID- 17085906 TI - [A case report of early gastric cancer with Paneth-like tumor cells]. AB - A 55-year-old man was given a diagnosis of grade 0IIc type gastric cancer of the midgastric posterior wall following endoscopy of the upper digestive tract, and subsequently underwent distal gastrectomy. Tumor cells resembling Paneth cells were occasionally observed in differentiated adenocarcinoma that had invaded the submucosal layer, the tumor was CD10-positive, and showed differentiation to complete intestinal metaplasia. In addition, p53-positive cells were also occasionally observed. Although complete intestinal metaplasia is associated with a low incidence of malignant transformation, this case was considered to be different from complete intestinal metaplasia, on which a mutation of the gene of p53 is involved in the early period of carcinogenesis. PMID- 17085907 TI - [A case of H. pylori and API2-MALT1 gene-negative gastric mucosa-associated lymphoid tissue lymphoma with a carcinoma-like signet-ring cell lymphoepithelial lesion]. AB - A 60-year-old woman presented in February 2003 with an ulcer on the lesser curvature of the anglus. The endoscopic biopsy specimens showed epithelial signet ring cell associated with lymphoid infiltration, suggesting a diagnosis of gastric cancer. Histopathological examination confirmed the diagnosis of low grade B-cell lymphoma of MALT with epithelial signet-ring cell lymphoepithelial lesion, which was negative for H. pylori and t (11;18) (q21;q21) translocation (API2-MALT1 gene). This case was treated with H. pylori eradication and additional radiation therapy, and the tumor was disappeared. PMID- 17085908 TI - [A case of intestinal intussusception caused by multiple lipoma diagnosed preoperatively]. AB - A 45-year-old man presented with intermittent abdominal pain for a month. Intestinal intussusception was diagnosed by the findings of abdominal ultrasonography and computed tomography. Intussusception was reduced by barium enema. Based on magnetic resonance imaging, we diagnosed intestinal intussusception due to multiple lipoma. At laparotomy, after successful reduction of the intussusception with Hutchinson's maneuver, an ileoceal resection was performed. Intussusception in adults is relatively rare and may be difficult to diagnose preoperatively. In the diagnosis of this disease, abdominal echo, CT, and MRI are useful. PMID- 17085909 TI - [Acute hemorrhagic colitis induced by the neuraminidase inhibitor oseltamivir]. AB - A 23-year-old woman had lower abdominal pain, diarrhea and bloody stool was admitted and given a diagnosis of influenza B. Her home doctor had started treatment by neuraminidase inhibitor (oseltamivir) the previous day. Colonoscopic examination revealed an area of hemorrhage and erosion in the left transverse colon. After halting oseltamivir treatment these symptoms disappeared and her colonoscopic findings improved. A drug-induced lymphocyte stimulation test was positive for oseltamivir. This case is the first reported case of acute hemorrhagic colitis induced by oseltamivir. PMID- 17085910 TI - [A case of gastrointestinal stromal tumor with liver and bone metastases effectively treated with radiofrequency ablation and imatinib mesylate]. AB - A 58-year-old man was admitted because of perforation of the small intestine by a gastrointestinal stromal tumor (GIST). First, the small intestine including a GIST was resected; and then 2 month later, a part of the liver (S5) conforming to metastatic lesion was surgically removed. Twelve months later, another liver metastases was found, and surgical treatment was recommended; but the patient requested non-surgical therapy, so a radiofrequency ablation (RFA) was successfully performed. After that, recurrence of liver metastasis was not observed, but another metastasis was observed on the fifth lumbar vertebra; so administration of imatinib mesylate was started. 28 months after the initial administration the metastatic liver lesion was still invisible, and the bone metastatic lesion had not grown. The patient is alive with good performance status. This report shows that multi-modality therapy by surgery, RFA and imatinib mesylate was effective for liver and bone metastases of GIST. PMID- 17085911 TI - [Two cases of pancreatic pseudocyst with bleeding]. AB - We encountered 2 patients (No. 1 and 2) with pseudocyst hemorrhage of the pancreas. Patient No. 1, who presented with hemorrhagic shock due to rupture of a splenic aneurysm, was evaluated as a responder based on the response to the initial transfusion, and emergency TAE (transcatheter arterial embolization) was performed, which proved to be a successful life-saving measure. In Patient No. 2, also judged to be a responder, angiography was conducted and the course could be observed, because the hemorrhage was localized in the cyst. These results indicate that it is important to promptly select treatment policies based on the hemodynamic responses to the initial transfusion in cases with cystic hemorrhage of the pancreas. PMID- 17085912 TI - [A case of pancreatic solid-pseudopapillary tumor without cystic component in an elderly man]. AB - We encountered a solid pseudopapillary tumor (SPT) without any cyst component in an elderly man. A literature survey of SPT without cystic component suggested that the size and vascularity of a tumor were related to lack of cystic component. The prognosis of this disease was comparatively good, but elderly or middle aged case and cases without capsule seem to have high malignant potential. PMID- 17085913 TI - Possible broad impacts of long work hours. AB - The paper summarizes research linking long work hours to a wide range of risks to workers, families, employers, and the community. The risks are theorized to stem from less time to recover from work, longer exposure to workplace hazards, and less time to attend to non-work responsibilities. Risks to workers include sleep deprivation, poor recovery from work, decrements in neuro-cognitive and physiological functioning, illnesses, adverse reproductive outcomes, and injuries. Risks to families include delayed marriages and child bearing, and obesity in children. Risks to employers include reduced productivity and increases in workers errors. Mistakes by fatigued workers have broad reaching impacts to the community: medical errors, automobile crashes with other drivers on the road, and industrial disasters that damage the environment. PMID- 17085914 TI - Health problems due to long working hours in Japan: working hours, workers' compensation (Karoshi), and preventive measures. AB - Late in the 1970s, serious social concern over health problems due to long working hours has arisen in Japan. This report briefly summarizes the Japanese circumstances about long working hours and what the Government has achieved so far. The national statistics show that more than 6 million people worked for 60 h or more per week during years 2000 and 2004. Approximately three hundred cases of brain and heart diseases were recognized as labour accidents resulting from overwork (Karoshi) by the Ministry of Health, Labour and Welfare (MHLW) between 2002 and 2005. Consequently, the MHLW has been working to establish a more appropriate compensation system for Karoshi, as well as preventive measures for overwork related health problems. In 2001, the MHLW set the standards for clearly recognizing Karoshi in association with the amount of overtime working hours. These standards were based on the results of a literature review and medical examinations indicating a relationship between overwork and brain and heart diseases. In 2002, the MHLW launched the program for the prevention of health impairment due to overwork, and in 2005 the health guidance through an interview by a doctor for overworked workers has been enacted as law. Long working hours are controversial issues because of conflicts between health, safety, work-life balance, and productivity. It is obvious that we need to continue research regarding the impact on worker health and the management of long working hours. PMID- 17085915 TI - Study on a model for future occupational health: proposal for an occupational health service model in Japan. AB - The Study Model for Future Occupational Health (funded by a research grant from the Ministry of Health, Welfare and Labor) is a joint research project involving various organizations and agencies undertaken from 2002 to 2004. Society has undergone a dramatic transformation due to technological developments and internationalization. At the same time a low birth rate and an aging population have resulted in an increase in both the percentage of workers experiencing strong anxiety and stress in relation to their jobs and the working environment and the number of suicides. As a natural consequence, occupational health services are now expected to provide EAP, consulting and other functions that were formerly considered outside the realm of occupational health. In consideration of this background, the present study propose the following issues to provide a model for future occupational health services that meet the conditions presently confronted by each worker. 1. How to provide occupational health services and occupational physicians' services: 1) a basic time of 20 minutes of occupational health services per year should be allotted to each worker and to all workers; 2) the obligatory regulations should be revised to expand the obligation from businesses each with 50 or more employees under the present laws to businesses each with 30 or more employees. 2. Providers of occupational health services and occupational physicians' services: (1) reinforcement of outside occupational health agencies; (2) fostering occupational health consultant firms; (3) development of an institute of occupational safety and health; (4) support of activities by authorized occupational physicians in the field; (5) expanding of joint selection of occupational physicians including subsidy increase and the extension of a period of subsidy to five hears; (6) licensing of new entry into occupational health undertaking. 3. Introduction of new report system: (1) establishment of the obligation to submit reports on risk evaluation and improvement measures; (2) establishment of the obligation to prepare a report on results of medical examinations in all sizes of businesses. 4. Introduction of a merit system into businesses in establishment of a new system: the application of the special merit system of the workers' compensation insurance shall be revised to add occupational health activities, cover business with 20 or more to 100 or less employees and expand the period of application for three years under the present laws to five years. 5. Ensuring of international coordination: harmonization of standards of individual countries for occupational health and safety; thorough (1) ensuring of international agreement on high-level specialist qualifications; (2) mutual recognition of qualifications of occupational physician, nurse, occupational hygienist, ergonomist, and counselor; (3) preparation of guidelines for occupations relating to occupational health businesses. PMID- 17085916 TI - Lung disorders of workers exposed to rush smear dust in China. AB - The aim of this study was to evaluate the lung disorders of the workers exposed to rush smear dust. A cross sectional study was carried out on 1,709 current workers (788 male, 921 female) in 80 factories. All subjects were asked by questionnaire, and health examination including chest X-ray was conducted for 661 workers in 35 factories. Lung function test was also examined for 119 non-smoking males among 661 subjects. Dust samplings were collected and total and respirable dust concentrations at 127 spots in 35 factories were measured. The geometric mean dust concentration in the workshops was up to 20.00 mg/m(3), and the geometric mean respirable dust concentration reached 8.22 mg/m(3). The mean quartz concentration of accumulated dust was 29.2%. The prevalence of radiographic small opacities profusion category > or = 1/0, according to the ILO 1980 Classification System, was 2.6% among 661 employees. One worker was found to have pneumoconiotic findings of 2/2 profusion accompanied with large opacity. The prevalence of pneumoconiosis (1/0 or more) correlated with cumulative dust exposure (r=0.192, p<0.0001). The similar relationship was found between the prevalence rate of cough or sputa and worksite dust concentration. In non smokers, a positive association was found between the prevalence of cough and occupational exposure duration (r=0.080, p=0.004). Approximately 19.3% and 34.5% of employees suffered from respiratory impairment for FVC and FEV1.0, respectively. This is the first report of "rush" pneumoconiosis in China. Rush mat workers were found to be at high risk for pneumoconiosis, a preventable disease. Our results showed a dose-response relationship between rush-mat dust level and the prevalence of pneumoconiosis. Similar relationship between the prevalence of cough and sputum and the work duration was found for non-smoking workers but not for smoking workers. PMID- 17085917 TI - Sleepiness and head movements. AB - Sleepiness in working life is critical and strongly associated to work related accidents. The relationship between sleepiness and head movements is poorly investigated. The pattern of head movements over time was investigated in a laboratory study with ten subjects either sleep-deprived or rested. Head movements were obtained by an inclinometer placed on the subject's forehead, and the recording was continuous. Results show that subjects when sleep-deprived moved their head more and had more extreme head movements compared to when rested. An increase of the velocity and the number of extreme head movements over time were noted when the subjects were sleep-deprived and when rested. The increase of head movements was more linear over time in the rested condition, whereas in sleep-deprived conditions most of the increase appeared during the first hour. No significant differences of between forward-backward movements and left-right movements could be found. When rested, the changes in head movements correlated with ratings of sleepiness, EEG activity, and heart rate variability. Head movements can be a used as an indicator of sleepiness. PMID- 17085918 TI - Cold-provocation testing for the vascular component of hand-arm vibration syndrome in health surveillance. AB - The aim was to investigate whether the use of infra-red thermography (I-R) and measurement of temperature gradients along the finger could improve the diagnostic accuracy of cold-provocation testing (15 degrees C for 5 min) in vascular hand-arm vibration syndrome (HAVS). Twenty-one controls and 33 individuals with stages 2/3V HAVS were studied. The standard measurement of time to rewarm by 4 degrees C (T4 degrees C) and temperature gradients between the finger tip, base and middle (measured using I-R) were calculated. Receiver Operating Characteristics (ROC) analysis to distinguish between the two groups revealed that for T4 degrees C the area under the ROC curve was not statistically significantly different from 0.5 (0.64 95% confidence interval 0.49-0.76). The difference between the tip and middle portion of the finger during the sixth minute of recovery was the most promising gradient with an area of 0.76 (95% confidence interval 0.62-0.87), and sensitivity and specificity of 57.6% and 85.7% respectively. However, this was not significantly different from that for the time to rewarm by 4 degrees C. In conclusion, the cold-provocation test used in this study does not appear to discriminate between individuals with stage 2/3V HAVS and controls and this is not improved by the measurement of temperature gradients along the fingers using I-R. PMID- 17085919 TI - A study on compliance to hearing conservation programme among industries in Negeri Sembilan, Malaysia. AB - Noise is one of the hazards faced by workers. A cross-sectional study was conducted among industries in Negeri Sembilan with the objective to assess their compliance to Hearing Conservation Programme (HCP). The other objectives of this study were to determine the factors influencing it and to show the industries' compliance to each element of the programme. It was also to identify the association between compliance to HCP and the prevalence of hearing impairment and standard threshold shift. Data for this study were collected using questionnaires sent by mail and also the results of the latest audiometric tests. A total of 167 industries were analysed for this study. It was found that 41.3% of these industries fully complied to the programme. It was also found that the industries preferred to provide hearing protection device (92.8%) and least complied to noise control (61.1%). There were significant associations (p<0.05) between compliance and number of employees, status of ownership and the presence of officer in charge of hearing conservation programme. Having at least 150 employees actually raised the compliance to HCP in two folds (beta = 0.717, OR = 2.048, C.I 95% = 1.063 to 3.944). The prevalences of hearing impairment and standard threshold shift were 23.9% and 5.2% respectively. There was no significant association between the prevalence for hearing impairment and compliance to HCP. The prevalence for standard threshold shift was inversely related to compliance. This study showed that compliance percentage need to be improved as an effort to prevent the hearing problems among workers exposed to noise. PMID- 17085920 TI - Decreases in CD8+ T, naive (CD4+CD45RA+) T, and B (CD19+) lymphocytes by exposure to manganese fume. AB - To examine the effects of exposure to manganese (Mn) on the cellular and humoral immune system in men, T lymphocyte subpopulations, B (CD19+) lymphocytes, natural killer (NK) cells, and serum immunoglobulins (i.e., IgG, IgA and IgM) together with total T (CD3+) lymphocytes and total lymphocytes were measured in blood samples from 21 welders mainly exposed to Mn fume with blood Mn (BMn) concentrations of 0.6-2.3 (mean 1.4) microg/dl and 21 healthy controls working in the same factory (BMn concentrations: 0.7 to 1.7, mean 1.1 microg/dl). The workers engaged in welding for 6 to 36 (mean 17) yr. All the study subjects were divided into 3 equally sized groups (n=14 for each group) according to BMn concentrations. Numbers of CD8+ T, total T (CD3+), B (CD19+), and total lymphocytes were significantly lower in high-BMn group than those in low-BMn group; the numbers of CD8+ T lymphocytes were significantly lower in moderate-BMn group compared to low-BMn group. After adjusting for age and smoking, significant inverse correlations between BMn concentrations and CD4+CD45RA+ T, CD4+ T, CD8+ T, CD3+ T, and total lymphocytes were found. We conclude that T lymphocytes, especially CD8+ and CD4+CD45RA+ T lymphocytes, as well as CD19+ B lymphocytes are affected by exposure to Mn fume. PMID- 17085921 TI - Mental ability and psychological work performance in Chinese workers. AB - This study was to explore the relationship among mental ability, occupational stress, and psychological work performance in Chinese workers, and to identify relevant modifiers of mental ability and psychological work performance. Psychological Stress Intensity (PSI), psychological work performance, and mental ability (Mental Function Index, MFI) were determined among 485 Chinese workers (aged 33 to 62 yr, 65% of men) with varied work occupations. Occupational Stress Questionnaire (OSQ) and mental ability with 3 tests (including immediate memory, digit span, and cipher decoding) were used. The relationship between mental ability and psychological work performance was analyzed with multiple linear regression approach. PSI, MFI, or psychological work performance were significantly different among different work types and educational level groups (p<0.01). Multiple linear regression analysis showed that MFI was significantly related to gender, age, educational level, and work type. Higher MFI and lower PSI predicted a better psychological work performance, even after adjusted for gender, age, educational level, and work type. The study suggests that occupational stress and low mental ability are important predictors for poor psychological work performance, which is modified by both gender and educational level. PMID- 17085922 TI - A field evaluation method for assessing whole body biomechanical joint stress in manual lifting tasks. AB - Work-related musculoskeletal injuries are often associated with overexertion of the body at work. The manual materials handling activity of lifting is a major source of work-related musculoskeletal disorders. Biomechanical evaluation offers useful information about the physical stress imposed on the worker's body joints; however, biomechanical analysis is usually tedious and complex. For evaluation purpose, the biomechanical method needs to be easy to apply in a field environment. Manual lifting occurs as one of the most common manual materials handling tasks in the workplace. A biomechanical evaluation method was developed based on the ratio of joint moment to joint capacity. The method was applied to evaluate the physical stress of manual lifting in truck loading jobs using a nine link whole body joint model. Thirty eight industrial tasks were evaluated using the developed joint moment ratio. The moment ratio was compared with subjectively rated body discomfort, overall workload, and the NIOSH lifting index. The moment ratio was found to have a high correlation with the NIOSH lifting index. The biomechanical method can be used with relatively simple equipment and procedure which may be suitable for on-site ergonomic evaluation. PMID- 17085923 TI - Mental health status of municipal solid waste incinerator workers compared with local government office workers. AB - Recently in Japan dioxin problem of municipal solid waste incinerator (MSWI) became social issue. The news spread all around Japan and induced fear that workers at incinerators would suffer from cancer or other serious illness induced by the exposure to dioxins. Authors were interested in the effect of this stressful event occurred to the workers and intended to evaluate mental health status of MSWI workers compared with office workers. Subjects were male workers from two MSWI plants and a local government office; 20 government office workers who were engaging in health administration and 55 MSWI workers. Subjects were interviewed about their age, educational carrier, and working schedule. POMS and GHQ30 were used to evaluate mood status of subjects. There were differences in mood state between the two occupational groups. POMS showed that Tension-Anxiety, Depression-Dejection, and Fatigue levels were high in the health administration worker group. GHQ30 showed that General Illness, Social Dysfunction, and Anxiety and Dysphoria state were deviated to abnormal in the health administration worker group. General mental health status evaluated by GHQ30 score was also deviated to abnormal in the office worker group. Our results showed that mental health status of health administration workers was less healthy compared with MSWI workers. This meant that the stress of MSWI workers enhanced by the fear that they might have been exposed to dioxin did not exceed the stress the health administration workers usually had suffered from. PMID- 17085924 TI - Relationship between blood lead level and work related factors using the NIIH questionnaire system. AB - Over an 11-yr period (1990-2000), a questionnaire survey on work environmental management and environmental improvement was conducted on 259 lead-handling factories and 7,623 subjects. Labour Inspection Offices identified these factories as requiring environmental improvement, or possessing a desire to improve their working environment. We analyzed factors affecting blood lead levels (PbBs). These factors were gender, age, employment duration, factory size, work environment control (WEC) class, and job categories. The PbB of men was found to be higher than that of women, and may be due to the differences in job distribution. PbB increased along with increasing age and employment duration. PbB declined as the factory size increased. The odds ratio (OR) of PbB higher than 20 microg/dl according to factory size was significantly high even after adjusting for WEC class. This demonstrates that not only the working environment but also safety management was poorer among small-scale factories that large scale factories. The rise of PbB along with the increase of WEC class confirmed that the results of work environment measurement are correlated with individual exposure levels. The risk of having a 20 microg/dl or higher PbB was different for various lead handling jobs. Smelting or refining lead had the highest risk for lead exposure while painting or baking had the lowest risk. As our study population was not a randomly selected sample, we are unable to generalize our results for workers across Japan. However, we were able to indicate which jobs pose a high-risk and the effectiveness of using the work environment control class as an index of worksite environment levels. PMID- 17085925 TI - Occupational asthma after withdrawal from the occupational allergen exposure. AB - Occupational asthma is characterised by airway inflammation, variable airflow limitation and airway hyperresponsiveness related causally to work. The aim of the study was to ascertain whether in patients with occupational asthma findings persist after withdrawal from occupational allergen exposure. A group of 37 patients with occupational asthma and a control group of 19 persons were examined. Results in asthmatics obtained during the first visit when occupational asthma was acknowledged, were compared with recent results about 6.5 yr on average after elimination of occupational allergen exposure. Recent findings in occupational asthma patients were compared with the control group. In patients with occupational asthma, no significant differences were found between results obtained at the first and recent visit. Symptoms of asthma persisted in as much as 86.5% of patients. During recent examination there were more positive results in asthmatic patients comparing with the control group in histamine challenge (61.3 vs. 5.3%, p<0.01), eosinophile cationic protein (41.7 vs. 5.3%, p<0.05), prick tests (45.9 vs. 10.5%, p<0.05). Positive results of the present histamine challenge test and elevated eosinophils in sputum were more frequent (p<0.05) in patients with occupational asthma due to high molecular weight allergens than to low molecular weight allergens. PMID- 17085926 TI - Dissolution of functional materials and rare earth oxides into pseudo alveolar fluid. AB - The dissolution rates of rare earth oxides and two types of rare earth containing functional materials into water, saline solution, and Gamble's fluid were measured in order to evaluate the biological effects of rare earth-containing functional materials. The tested materials were yttrium, lanthanum, cerium and neodymium oxides, and neodymium-boron-iron magnet alloy (NdBFe) and lanthanum mish-metal-nickel-cobalt (LmNiCo) hydrogen-containing alloy. The dissolution rates of the rare earth oxides were very low, resulting in concentrations of rare earth elements in the test solutions of the order of ppb. In the most extreme case, Gamble's fluid dissolved 1,400 times more of the rare earth oxides than pure water. Fairly high concentration of neodymium were found in the dissolving fluids, which means that trace neodymium present as an impurity in each rare earth oxide dissolved preferentially. For yttrium oxide, the ratio of neodymium to yttrium that dissolved in the saline solution was greater than 78,000 to 1, taking into account the amount of each that was originally present in the yttrium oxide. PMID- 17085927 TI - Perceptions of work and family assistance and the prevalence of lower back problems in a South African manganese factory. AB - An analytical cross-sectional epidemiological study was carried out among 109 manganese plant workers to examine the prevalence and association between lower back problems (LBP) and family and workplace related psychosocial risk factors. Outcome (LBP) was defined using a guided questionnaire and a functional rating index. Exposure to family and workplace related psychosocial risk was determined using the Occupational Risk Factor (ORFQ) and APGAR questionnaires for work and family support. Using inclusive and stringent definitions for perceived LBP, point prevalence was 37.6% and 29.4%, respectively. Only 8 cases of LBP were, however recorded officially over a 7-yr period (1996-2003). Multivariate analyses indicated a high, but non-significant odds ratio (OR) for negative perceptions of workplace support OR 3.29 (CI 0.95-11.30). A positive, non-significant, association for negative perceptions of family support (2.56; CI 0.69-9.52) and a protective, but non-significant, association for control over the order and pace of working tasks (OR 0.40; CI 0.12-1.35) was found. These findings together with the under-reporting, leads one to conclude that work hardening and a cultural ethos of non-complaining among manual labourers, moderates the association between LBP and the aetiological factors studied. PMID- 17085928 TI - Effects of in utero and lactational exposure to di(2-ethylhexyl)phthalate on somatic and physical development in rat offspring. AB - Di(2-ethylhexyl)phthalate (DEHP) has been reported to act as an antiandrogen and to affect the reproductive organs and accessory genital glands. Thus, to assess the reproductive toxicity of DEHP it is important to examine both its adverse effects on the development of offspring following maternal exposure and its effects on sexual function and fertility. In the present study, we examined whether in utero and lactational exposure to DEHP affects postnatal somatic growth of offspring in the rat. Pregnant females were orally administered various doses of DEHP (0, 25, 100 or 400 mg/kg body weight/day) from gestational day (GD) 6 through postnatal day (PND) 20. There were no significant changes in body weight, body length, tail length, or the weight of individual organs between the control and DEHP-treated groups. Somatic hormonal parameters were the same for all DEHP doses. These findings suggest that in utero and lactational exposure to various concentrations of DEHP has very little effect on postnatal development or endocrine and physical status of male and female rat offspring under the experimental conditions of the present study. PMID- 17085929 TI - Multimedia for occupational safety and health training: a pilot study examining a multimedia learning theory. AB - Occupational diseases are a significant problem affecting public health. Safety training is an important method of preventing occupational illness. Training is increasingly being delivered by computer although theories of learning from computer-based multimedia have been tested almost entirely on college students. This study was designed to determine whether these theories might also be applied to safety training applications for working adults. Participants viewed either computer-based multimedia respirator use training with concurrent narration, narration prior to the animation, or unrelated safety training. Participants then took a five-item transfer test which measured their ability to use their knowledge in new and creative ways. Participants who viewed the computer-based multimedia trainings both did significantly better than the control group on the transfer test. The results of this pilot study suggest that design guidelines developed for younger learners may be effective for training workers in occupational safety and health although more investigation is needed. PMID- 17085930 TI - Inhibitory effect of ethylene glycol monoethyl ether on rat sperm motion. AB - Ethylene glycol monoethyl ether (EGEE) is known to have testicular toxicity. To elucidate whether EGEE has any effect on sperm motion, especially in the case of short time exposure, we conducted a series of in vivo experiments with rats, as well as an in vitro study with rat sperm. Sperm from cauda epididymides and spermaducts was analyzed for the change in motion with a Hamilton-Thorne Sperm analyzer. Administration of EGEE at 600 mg/kg/d for five weeks significantly decreased total and progressive motility of sperm to 15-30% of controls, in both the cauda epididymis and the spermaduct. The time-course experiment using a single dose of 1,000 mg/kg showed that damage to sperm motion was evident at 12 24 h after EGEE administration. Addition of EGEE to the medium of sperm had no effect on its motion, but the metabolite ethoxyacetic acid showed a significant inhibitory effect. These results suggest that besides its toxicity to spermatogenesis, the metabolite of EGEE may also directly affect the motion of mature sperm. PMID- 17085931 TI - Arm pain and daytime sleepiness among nursing home employees. AB - We investigated the relationship between musculoskeletal disorders and sleep problems among 98 employees (79 women) at three nursing homes. Self-reported data were collected regarding pain in the neck, shoulders, arms, legs, and low back, sleep disturbances, daytime sleepiness (Epworth Sleepiness Scale), and the level of workload on the hands, legs, and low back by type of care. Pain in the arms was significantly associated with less difficulty initiating sleep, fewer symptoms of insomnia, and a higher level of daytime sleepiness. After adjusting for age and gender, only the association between arm pain and daytime sleepiness remained significant (Odds Ratio 6.70, 95% Confidence Interval 1.40-31.97). Participants with both arm pain and daytime sleepiness showed significantly greater levels of workload in some kinds of care in a systemic manner than counterparts without either complaint. These findings suggest that arm pain is associated with elevated sleep propensity/fatigue in nursing home work. PMID- 17085932 TI - Analysis of human proteins that have an affinity to heavy metals by metal chelating column chromatography. AB - To clarify the molecular basis of toxicities of industrial chemicals, it is demanded to develop appropriate methods whereby their cellular target molecules can be directly identified. In the present study, we focused on target proteins of heavy metals and established the method to detect them using a combination of metal-chelating column chromatography and a subsequent analysis by electrophoresis. Protein samples prepared from HeLa cells were applied to the Zn- or Cd-chelating column, and the bound proteins were analyzed by SDS polyacrylamide gel electrophoresis followed by either silver staining, or fluorography when using radiolabel protein samples. Among several protein species trapped in the columns, a 36-kDa protein apparently has an affinity to both Zn and Cd, indicating the possibility that Cd can exchange essential Zn on this protein. These results suggest that the established method is useful for the target protein screening and further analyses of separated proteins. PMID- 17085933 TI - Regulation of proopiomelanocortin gene transcription during single and repeated immobilization stress. AB - We have previously reported that repeated immobilization produces persistent activation of the hypothalamic-pituitary-adrenocortical axis in rats. In an attempt to assess whether any adaptational responses occur at the pituitary level, we examined the detailed time courses of proopiomelanocortin (POMC) gene transcription in the anterior pituitary (AP) in comparison with those of corticotropin-releasing hormone (CRH) gene transcription in the hypothalamic paraventricular nucleus (PVN) during single and repeated immobilization using both intronic and exonic probes. During single immobilization, there was a robust and rapid increase in both CRH heteronuclear RNA (hnRNA) in the PVN and POMC hnRNA in the AP, together with a slower increase in CRH mRNA, but no significant increase in POMC mRNA. Single immobilization also caused significant increases in the plasma concentrations of both ACTH and corticosterone. Daily immobilization for 6 days increased the basal levels of CRH hnRNA and CRH mRNA in the PVN and POMC mRNA in the AP. Both CRH hnRNA and POMC hnRNA responded rapidly to a final episode of acute immobilization on day 7, whereas the peak values of CRH hnRNA and POMC hnRNA after 15 min of the final stress were smaller than those during single immobilization. In contrast to single stress, CRH mRNA did not change significantly, whereas POMC mRNA robustly increased after the final immobilization on day 7. Plasma ACTH increased to a similar degree to single stress, but its initial increase at 5 min was significantly higher than that during single immobilization. The increase in the plasma corticosterone concentration was higher during final immobilization than during single stress. These results suggest that, in response to the hypothalamic drive during repeated immobilization stress, pituitary corticotrophs are capable of upregulating the basal and stress-induced POMC mRNA levels via increased efficiency of the posttranscriptional processing of the hnRNA and/or increased mRNA stability. PMID- 17085934 TI - Left ventricular diastolic function in hemodialysis patients: role of preload increase maneuver on tissue Doppler imaging evaluation. AB - OBJECTIVE: Tissue Doppler imaging (TDI) has recently been proposed as a relatively preload-independent method to evaluate left ventricular diastolic function. We sought to investigate the higher-accuracy of TDI to assess diastolic function in end-stage renal disease (ESRD) patients on hemodialysis (HD) associated with a preload increase maneuver. METHODS: Thirty-two consecutiveESRD patients (16 female, ages 48.8 +/- 17.5 years, 14 45 years old) were evaluated. Measurements of E, A velocities and the E/A ratio from transmitral inflow pulsed wave Doppler, and E', A' velocities and the E'/A' ratio from TDI were obtained 1 h before and 1 h after HD at baseline and with a preload increase maneuver. RESULTS: The E/A ratio changed significantly in all patients aged >45 before and after HD with the preload increase maneuver. The E'/A' ratio increased in all subjects with the preload increase maneuver before HD but did not change with the maneuver after HD in the euvolemic state in all patients. CONCLUSION: In ESRD patients on routine HD, TDI evaluation associated with a preload increase maneuver proved to be a more accurate method to identify diastolic dysfunction when the evaluation is performed in euvolemic patients after HD. PMID- 17085935 TI - Abnormal intracellular calcium handling underlying T-wave alternans and its hysteresis. AB - AIMS: To investigate the mechanism underlying T-wave alternans (TWA) and its hysteresis under ischemia conditions. METHODS: Transmembrane action potential (AP) from endocardial, M, and epicardial cells and monophasic AP (MAP) from four epicardial sites were recorded in ventricular wedge preparation and in isolated intact rabbit heart, respectively. The AP/MAP duration (APD/ MAPD), effective refractory period (ERP), activation time, and APD/MAPD restitution were determined under control and ischemia conditions. The effects of ryanodine (0.01 and 1 micromol x l(-1)) on TWA, and the effects of low extracellular Ca2+ and 4 aminopyridine on its hysteresis were studied. RESULTS: Ischemia shortened the APD/MAPD and effective refractory period of all recording sites symmetrically, except the APD of M cells, which shortened markedly. In the ischemia group, TWA was induced within a cycle length (CL) range from 160 to 250 ms, which corresponded to a diastolic interval region of 0-70 ms. In this diastolic interval region, the repolarization restitution curve was the steepest (slope > 1.0). All TWA were accompanied by repolarization alternations. Low concentration ryanodine (0.01 micromol x l(-1)) facilitated TWA, high concentration (1 micromol x l(-1)) abolished it. Alternans of calcium transient were observed in myocytes purfused with ischemia solution during rapid stimulation. Ryanodine (0.1 micromol x l(-1)) abolished alternans of calcium transient, and ryanodine (0.01 micromol x l(-1)) facilitated them. After 60 min pacing at a CL of 200 ms, TWA persisted until the initial several beats at a CL of 300 ms at which a TWA was exceptional. The suppression of hysteresis by low extracellular Ca2+ and 4-aminopyridine indicated an underlying role of the intracellular Ca2+ overload and transient outward current (I(to)). CONCLUSION: TWA is principally due to repolarization alternans, which is secondary to steep APD/MAPD restitution, and relates to intracellular calcium cycling. Hysteresis relates to intracellular Ca2+ overload and I(to). PMID- 17085936 TI - Clozapine-induced effuso-constrictive pericarditis. Case report and review of the literature. AB - Clozapine is a widely used antipsychotic medication and is effective against both positive and negative symptoms of schizophrenia. However, clozapine use should be monitored closely due to its side effects profile. The main side effects include cardiorespiratory symptoms, seizures and agranulocytosis. We report a case of effuso-constrictive pericarditis soon after being started on clozapine therapy, whose symptoms improved after discontinuation of clozapine. The literature is reviewed and importance of monitoring is discussed. PMID- 17085937 TI - Interaction of plasma homocysteine and thyroid hormone concentrations in the pathogenesis of the slow coronary flow phenomenon. AB - BACKGROUND AND OBJECTIVE: The slow coronary flow (SCF) phenomenon is an angiographic observation and a well-recognized clinical entity characterized by delayed opacification of vessels in a normal coronary angiogram due to reasons yet unclear. Thyroid hormones exert significant effects on plasma homocysteine (Hcy) levels and microvascular resistance. Recently, several investigators have consistently reported that elevation of the plasma Hcy level can severely disturb vascular endothelial function and play a role in the pathogenesis of SCF. Accordingly, we investigated the levels of plasma Hcy and thyroid hormones and their relationship in patients with SCF. METHOD: Forty-four patients with angiographically proven SCF (Group I) (mean age 55.5 +/- 10.4 years, 26 males) and 44 cases with normal coronary flow (NCF) pattern (Group II) (mean age 53.9 +/ 11 years, 22 males) with similar risk profiles were enrolled in the study. Coronary flow patterns of the cases were determined by the thrombolysis in myocardial infarction (TIMI) frame count method. The coronary TIMI frame counts were calculated separately for each coronary artery and their average was determined as the mean TIMI frame count for each subject. Serum levels of free tri-iodothyronine (fT3), free thyroxine (fT4), thyroid stimulating hormone (TSH) and Hcy were measured. Patients with thyroid disease or on medications with a potential to affect thyroid functions were excluded. RESULTS: There were no statistically significant differences between the groups concerning the demographic characteristics and major cardiovascular risk factors. Mean TIMI frame counts of SCF and NCF groups were 45.9 +/- 12 and 23.3 +/- 3.7, respectively. fT4 (ng/dl) and TSH (microIU/ml) levels of the two groups were similar (p > 0.05). The level of fT3, the active metabolite of the thyroid hormone family, was dramatically reduced in the SCF group when compared to the NCF group (2.3 +/- 0.2 vs. 3.0 +/- 0.3, p = 0.0001, respectively). Plasma Hcy levels of patients with SCF were found to be significantly higher than controls (12.2 +/- 4.9 vs. 8.5 +/- 2.9, p = 0.0001, respectively). Correlation analysis showed a significant negative correlation between the plasma fT3 and Hcy levels and the mean TIMI frame counts (r = -0.31, p = 0.003 vs. r = -0.66, p = 0.0001). Moreover, there was a significant positive correlation between the plasma Hcy levels and the mean TIMI frame counts (r = 0.58, p = 0.0001). Also, fT3 was the only significant determinant of the variance of Hcy in multiple regression analysis (r = -0.30, p = 0.005). CONCLUSION: fT3 levels were decreased and plasma Hcy levels were increased significantly in patients with SCF as compared to controls. This finding suggests that thyroid hormones and/or (?) a possible disturbance in their metabolism may be responsible for the elevated levels of plasma Hcy in patients with SCF and may play a role in the pathogenesis of the SCF phenomenon. PMID- 17085938 TI - Effectiveness of repeat enhanced external counterpulsation for refractory angina in patients failing to complete an initial course of therapy. AB - AIMS: This study examined the causes and results of retreatment of patients who failed to complete an initial 35-hour Enhanced External Counterpulsation (EECP) course. METHODS AND RESULTS: Data of 2,311 successive angina patients from the International EECP Patient Registry were analyzed, 86.5% completed their EECP course (Complete cohort). Of the 13.5% patients failing to complete the initial course (Incomplete cohort), 28.3% had repeat EECP within 1 year vs. 10.1% of the Complete group. The predictors of failure to complete the initial course of EECP were: female gender, heart failure, use of angiotensin-converting enzyme inhibitors or angiotensin receptor blockers, and use of nitroglycerin. For the Complete group, 83.4% had a reduction of at least one Canadian Cardiovascular Society (CCS) class after their initial EECP course, vs. 21.7% in the Incomplete group (p < 0.001). After repeat treatment, 66.2% of the Incomplete group achieved at least one CCS class reduction vs. 69.4% of the Complete group (p = NS) undergoing retreatment. The independent predictors for those who return to successfully complete their second course were patients who stopped their first course because of clinical events, and candidacy for coronary artery bypass grafting at the time of initial treatment. CONCLUSION: The results of retreatment of those who failed to complete their initial EECP course were comparable to those who completed their initial treatment, with similar reductions of CCS angina class. PMID- 17085939 TI - Effects of metoprolol on sympathetic remodeling and electrical remodeling at infarcted border zone after myocardial infarction in rabbits. AB - BACKGROUND: The findings of sympathetic remodeling and its electrophysiological implications force us to rerecognize the drugs presently used. The aim of this study was to investigate the effects of metoprolol on sympathetic remodeling and electrical remodeling at the infarcted border zone (IBZ) after myocardial infarction (MI). METHODS: Forty rabbits were randomly assigned into two groups: MI group (n = 20), ligation of the anterior descending coronary; Metoprolol group (n = 20), ligation of the anterior descending coronary and administration of oral metoprolol 5 mg/kg/day. Eight weeks after surgery, transmural dispersion of repolarization (TDR) at baseline, TDR and difference of TDR (deltaTDR) during sympathetic nerve stimulation were measured at the IBZ. The distribution and densities of growth associated protein 43 and tyrosine hydroxylase positive nerves at the IBZ were detected with immunohistochemical techniques. RESULTS: The study was completed in the 36 surviving animals (18 rabbits in each group). The densities of growth associated protein 43 and tyrosine hydroxylase positive nerves in the Metoprolol group (2,550 +/- 554 and 1,779 +/- 458 microm2/mm2, respectively) were lower than in the MI group (3,217 +/- 589 and 2,616 +/- 528 microm2/mm2, respectively; both p < 0.01). TDR at baseline, TDR and deltaTDR during sympathetic nerve stimulation were shorter in the Metoprolol group than in the MI group (p < 0.01 for all). CONCLUSION: Metoprolol can inhibit sympathetic remodeling and electrical remodeling at the IBZ after MI. The association of metoprolol with improved electrical remodeling may be partly related to the inhibition of sympathetic remodeling. PMID- 17085940 TI - Does the EGFR and VEGF expression predict the prognosis in colon cancer? AB - BACKGROUND/AIM: Epidermal growth factor receptor (EGFR) and vascular endothelial growth factor (VEGF) are frequently encountered with aggressive tumor phenotype and poor prognosis, but the relationship between EGFR/VEGF expression and survival remains unclear. The aim of our study was to further investigate the prognostic value of EGFR and VEGF expression in colon cancer. MATERIALS AND METHODS: The pathological specimens of 60 colon carcinoma patients were retrospectively evaluated and grouped according to EGFR and VEGF staining intensity and percentage of stained neoplastic cells. A final score was assigned to each case by multiplying percentage and staining score. The patients were stratified into the following categories: negative (score 0), low expression (score 1 or 2), and high expression (score 4). The remaining patient data were filtered out from the institutional cancer database. RESULTS: The mean survival time was 28.93 +/- 14.1 (range 2-52) months in the EGFR-negative group, 23.92 +/- 14.0 (range 6-46) months in the group with a low EGFR expression, and 17.00 +/- 12.8 (range 10-40) months in the group with a high EGFR expression. The median survival time was 27.50 +/- 14.7 (range 4-52) months in the VEGF-negative group, 29.33 +/- 12.8 (range 6-48) months in the group with a low VGEF expression, and 14.50 +/- 14.2 (range 2-40) months in the group with a high VGEF expression. The expression of EGFR and VEGF was not an independent factor that affects survival. CONCLUSIONS: The EGFR and VEGF expression rates of colon tumors do not predict the survival. In addition, the EGFR expression in the primary tumor was not predictive of metastatic lymph nodes. The prognostic value of EGFR/VEGF staining may be further questioned. PMID- 17085941 TI - Microanatomy of milk ducts in the nipple. AB - The aim of this study was to determine number and diameter of milk ducts in the nipple and to investigate the possible influences of age, breast weight, and diameter of the nipple on the number of ducts. Two hundred and twenty-six carcinoma mastectomy specimens were weighed and the nipple diameters measured. The number of ducts was counted in histological cross sections. Mean diameter of the nipple and mean breast weight were 13.9 mm and 844.6 g, respectively. There was a small but statistically significant positive correlation between nipple diameter and number of milk ducts (rho = 0.158; p = 0.01), but no correlation with breast weight. The mean number of ducts in the nipple duct bundle was 17.5. This is significantly higher than the number of ducts reported to open on the nipple surface. This discrepancy could reflect duct branching within the nipple or the presence of some ducts which do not reach the nipple surface. Smaller breast ducts (diameter < 0.5 mm) represent nearly 50% of the nipple ducts and could be a challenge to the ductoscopy technology. PMID- 17085942 TI - MTHFR 677C-->T and 1298A-->C polymorphisms: evaluation of maternal genotypic risk and association with level of neural tube defect. AB - BACKGROUND: Neural tube defects (NTDs) are common birth defects (1 in 1,000) leading to significant morbidity and mortality. Periconceptional folic acid supplementation helps in prevention of 70% of NTDs. Recently, polymorphisms in genes encoding enzymes of the folate pathway have been implicated in causation of NTDs. Since the closure of neural tube occurs at multiple sites, the etiology of defect at different sites may be different - which explains the failure of folic acid supplementation to prevent all NTDs. METHODS: Molecular analysis of methylenetetrahydrofolate reductase polymorphisms was carried out using polymerase chain reaction and restriction enzyme digestion. We studied the association of these polymorphisms in mothers with a previous child with NTD and further refined the risk by stratification based on level of defect. RESULTS: The frequency of 677C-->T homozygotes was higher in mothers with a previous child with NTD than the controls (OR = 1.6 (0.38-6.7), 95% CI, p = 0.72) but the difference was statistically insignificant. There was a significant difference in frequency of T alleles among mothers with a previous child with a 'lower' type of defect compared to controls (OR = 2.15 (1.13-4.1), 95% CI, p = 0.02). We did not find any significant association of 1298A-->C polymorphism with the level of NTDs. CONCLUSIONS: We conclude that in the North Indian population, the 677C-->T allele of the MTHFR gene may be associated with the occurrence of a lower type of NTD. This points towards the differential role of thermolabile MTHFR at different sites of neural tube closure. PMID- 17085943 TI - Pimecrolimus cream 1% in the treatment of lichen sclerosus. AB - BACKGROUND: Lichen sclerosus (LS) is a chronic inflammatory skin condition, which most commonly causes dysuria, pruritus and soreness of the vulval and perianal areas. Potent topical corticosteroids are used for the treatment of LS, but it is well known that they inhibit collagen synthesis and cause skin atrophy as a side effect. METHODS: The present pilot study evaluated the efficacy and safety of pimecrolimus cream 1% applied twice daily for up to 6 months in 29 women with severe LS. RESULTS: Of the 26 subjects who completed the follow-up period, 42% (11/26) were in complete remission with relief from itchiness, pain and inflammation. A 3.5-fold increase in type I collagen synthesis and a 7.5-fold increase in type III collagen synthesis of the affected areas was detected after 2 months of pimecrolimus treatment. There were no systemic adverse reactions, although mild local skin reactions were reported by 50% of the patients. Blood concentrations of pimecrolimus were checked in 10/26 patients (39%) and were undetectable in all cases. CONCLUSIONS: Patient-applied 1% pimecrolimus cream is safe and effective for the treatment of LS. PMID- 17085944 TI - Adrenarche in Prader-Willi syndrome appears not related to insulin sensitivity and serum adiponectin. AB - Prader-Willi syndrome (PWS) is a genetic disorder characterized by dysmorphic features, obesity, hypogonadism, hypotonia and mental retardation. Obesity has been linked to insulin resistance and the latter has also been associated with premature adrenarche. Since up to date a controlled study to investigate adrenarche and its hormonal regulation was lacking in PWS, our aim was to assess whether prepubertal PWS patients develop premature adrenarche and its relationship with markers of insulin sensitivity. Fourteen prepubertal children with PWS (6 M, 8 F) and 10 non-syndromal simple obese matched controls (5 M, 5 F) participated (mean age: 7.62 +/- 1.84 years). A fasting blood sample was obtained for adrenal and ovarian androgens, sex hormone binding globulin, insulin-like growth factor-I (IGF-I), insulin-like growth factor binding protein-1, leptin, adiponectin and a lipid profile. Thereafter an oral glucose tolerance test was performed. PWS patients were smaller at birth and a higher proportion displayed premature pubarche. No differences were found in testosterone, androstenedione, sex hormone binding globulin, free androgen index, homeostatic model assessment IR, 2-hour insulin, leptin or adiponectin levels. 17-hydroxyprogesterone and DHEAS levels however, were significantly higher in PWS. IGF-I levels were significantly lower in PWS and correlated significantly with height SDS (p < 0.05). In conclusion, a higher proportion of premature adrenarche in our PW patients was observed, which was not explained by differences in insulin sensitivity or plasma levels of adipokines and IGF-I. PMID- 17085945 TI - Independent regulation of Rap1 and mitogen-activated protein kinase by the alpha chain of Go. AB - Receptors coupled to G(i/o) proteins stimulate the mitogen-activated protein kinase (MAPK) cascade. The intracellular pathways linking the alpha chains of these G proteins to MAPK activation are not completely understood. One of the signaling molecules which has been suggested to act downstream of Galpha(i/o) is the small G protein Rap1. We investigated the role of Rap1 in MAPK stimulation by Galpha(o) in Chinese hamster ovary (CHO) cells. Our previous results have shown that in this cell system activated Galpha(o) strongly potentiates the MAPK response to the epidermal growth factor (EGF) receptor. Rap1 regulation was examined in cells transfected with Rap1 and wild-type Galpha(o) or the activated mutant Galpha(o)-Q205L. Immunocytochemical analysis detected both Rap1 and the Galpha(o) subunit at the plasma membrane as well as on perinuclear cytoplasmic vesicles. Expression of wild-type Galpha(o) had no significant effect on the levels of activated Rap1. In contrast, Galpha(o)-Q205L virtually abolished the activation of Rap1 induced by EGF. Further experiments showed that MAPK stimulation by EGF was greatly inhibited by expression of activated Rap1, suggesting that Rap1 inhibition could mediate the effect of Galpha(o) on the MAPK cascade. However, Galpha(o)-Q205L efficiently inhibited the activation of Rap1 induced by fibroblast growth factor (FGF). We have previously found that the ability of FGF to activate MAPK is not modified by Galpha(o). In addition, expression of the GAP protein RAP1GAPII blocked Rap1 activation without affecting EGF- or FGF-dependent MAPK stimulation. These findings provide evidence for independent regulation of Rap1 and MAPK by the G(o )alpha chain. PMID- 17085946 TI - Recent advances in radiation therapy for head and neck cancer. AB - The treatment of locally advanced or recurrent head and neck cancers has improved from single modality interventions of surgery and radiation therapy alone to include combined modality therapy with surgery, chemotherapy and radiation. Combined therapy has led to improved local control and disease-free survival. New developments in radiation oncology such as altered fractionation, three dimensional conformal radiotherapy, intensity-modulated radiotherapy, stereotactic radiosurgery, fractionated stereotactic radiotherapy, charged particle radiotherapy, neutron-beam radiotherapy, and brachytherapy have helped to improve this outlook even further. These recent advances allow for a higher dose to be delivered to the tumor while minimizing the dose delivered to the surrounding normal tissue. This article provides an update of the new developments in radiotherapy in the management of head and neck cancers. PMID- 17085947 TI - Morphologic alteration induced by short-term smoke exposure in rats. AB - BACKGROUND: It is a well-established fact that cigarette smoking causes degenerative, inflammatory, and respiratory diseases in humans. Because many factors such as air pollution and harsh working conditions can easily be eliminated in animal studies, we conducted this study to identify the effect of tobacco on rat trachea. METHODS: 24 male Wistar rats were divided randomly into an experimental and a control group. The experimental group of rats was exposed to cigarette smoke for 2 h each day over a duration of 60 consecutive days and the control group was treated in an identical fashion yet exposed only to room air. A morphometric study was performed on tracheal specimens taken from 22 rats (10 smoke-exposed rats and 12 control rats). RESULTS: Our results show that many of the morphological changes of the tracheal epithelium were found in the experimental group and significant quantitative differences were observed between the two groups. Loss of cilia, basal cell hyperplasia, goblet cell hyperplasia and an increased number of subepithelial inflammatory cells were observed by light microscopic examination of the trachea of experimental rats. We found very high levels of plasma thiocyanate after exposure to smoke in the experimental group, but no increase in the control group. CONCLUSIONS: The oxidants contained tobacco which could play an important role in the development of these structural and functional abnormalities in the trachea after smoke exposure. In addition, smoking can recruit inflammatory cells to the trachea. PMID- 17085948 TI - Contralateral normal ear after mastoid surgery: evaluation by otoacoustic emissions (mastoid drilling and hearing loss). AB - Mastoid drilling can cause transient hearing loss in the contralateral normal hearing ear. A study was designed to evaluate hearing in the contralateral normal ear before and after the mastoid surgery in a longitudinal manner and find out the duration of this temporary hearing loss. Twenty-two patients requiring mastoid surgery in their diseased ears, having contralateral normal ear were included. Pure tone audiometry and otoacoustic emissions (OAEs) were utilized for baseline evaluation. OAEs were repeated during the immediate postoperative period and daily up to the 6th postoperative day. The amplitudes of the OAEs of contralateral normal ears were found affected immediately after surgery and progressive improvement was detected with full recovery at 72-96 h. None of the patients had permanent deterioration in OAE amplitudes. The burs used during mastoid surgery can cause temporary hearing threshold changes in the contralateral ears. This adverse effect recovers spontaneously within 72-96 h postoperatively. PMID- 17085949 TI - Relationships among speech perception, self-rated tinnitus loudness and disability in tinnitus patients with normal pure-tone thresholds of hearing. AB - Exactly how speech perception and tinnitus perception are related remains unclear. This study investigated how tinnitus alone affects speech perception and the relationship between speech perception, tinnitus loudness, and tinnitus disability. The Mandarin Speech Perception in Noise Test (MSPIN), Tinnitus Loudness Scaling (TLS), and Tinnitus Handicap Inventory (THI) were utilized to assess 20 tinnitus patients with normal hearing. The tinnitus group had a significantly lower MSPIN score than the control group (p < 0.01). TLS and THI scores were strongly correlated (r(2): 0.534 approximately 0.627, p < 0.05). Correlations between MSPIN and TLS or THI scores were not significant. Tinnitus loudness correlated well with tinnitus-related disability. Neither tinnitus loudness nor disability was strongly correlated with speech perception. In noisy environments, tinnitus sufferers had significantly poorer ability to recognize speech than control subjects. PMID- 17085950 TI - The prognostic relevance of p16 inactivation in head and neck cancer. AB - The inactivation of the tumor suppressor gene p16 plays an important role in the development of malignant tumors. p16 loss can result from point mutations, loss of heterozygosity (LOH) or methylation of the promoter region. A total of 67 samples of tumor tissue from squamous cell carcinoma of the oral cavity, the pharynx and the larynx were analyzed for an inactivation of p16. The results of the molecular-biological investigations were correlated with the known clinical prognostic parameters after a follow-up period of approximately 3 years. Methylation of the promoter region and LOH were the main mechanisms of p16 inactivation. Point mutations presented as rare events. An inactivation of p16 did not have any statistical influence on tumor prognosis. Patients with a p16 gene inactivated by promoter methylation appeared to have a slightly lower tendency for local and regional recurrences. The inactivation of the tumor suppressor gene p16 plays a role in the carcinogenesis of head and neck cancer. PMID- 17085951 TI - Parapharyngeal abscess: comprehensive management protocol. AB - Parapharyngeal infections are rare, but they cause serious morbidity and mortality. Therefore, until now, the recommended treatment of parapharyngeal abscess has been early open surgical drainage. The purpose of this study is to review the clinical course and outcome of treatment in parapharyngeal abscess according to method of treatment. A prospective study was designed for parapharyngeal abscess in patients admitted for deep neck infection. During an 8 year period, from June 1994 to January 2003, 34 patients were enrolled. All had contrast-enhanced computed tomography (CT) imaging and confirmation of an abscess in the parapharyngeal space. All patients were treated with intravenous antibiotics. We treated 19 cases (conservative group) with antibiotics only or needle aspiration and 15 (surgical group) with intraoral or external drainage. The mean duration of hospitalization was 8.2 days in the conservative group and 11.6 days in the surgical group. There was no complication except mediastinitis in 1 case of the conservative group. Because of severe dyspnea, 5 patients required tracheotomy. Neck CT scan is a useful diagnostic tool to detect and establish the treatment plan of parapharyngeal abscess. Parapharyngeal abscess may, in some cases, respond to antibiotics, become localized to the parapharyngeal space and be treated conservatively with no need for early open surgical drainage. PMID- 17085952 TI - Vector-mediated delivery of bcl-2 prevents degeneration of auditory hair cells and neurons after injury. AB - OBJECTIVE: To test the hypothesis that bcl-2 prevents oxidative stress-induced apoptosis of auditory sensory cells in explants of the organ of Corti and dissociated cell cultures of the spiral ganglion. METHODS: Organ of Corti explants and dissociated spiral ganglion cell cultures obtained from 3-day-old (P3) rats or adult spiral ganglion cell cultures from 28-day-old (P28) rats were transduced with vectors containing a human bcl-2 gene. Cultures were then exposed to neomycin, cisplatin or subjected to withdrawal of neurotrophin supplementation. Outcome measures included hair cell and neuron counts, mitochondrial membrane potential and a histological measure of apoptosis. RESULTS: Expression of bcl-2 in the organ of Corti explants and neuronal cell cultures provided a significant level of protection against cell death. Bcl-2 expression in the organ of Corti explants also protected mitochondria from loss of membrane potential and blocked an early step in the commitment of hair cells to apoptosis. CONCLUSION: Expression of bcl-2 in cochlear tissues protects sensory cells from a variety of insults that have been demonstrated to damage the inner ear. PMID- 17085953 TI - The value of fine-needle aspiration cytology in salivary gland lesions, 1994 2004. AB - OBJECTIVE: Fine-needle aspiration cytology (FNAC) of the salivary gland is a commonly accepted, sensitive and specific technique in the diagnosis of both neoplastic and nonneoplastic lesions of the salivary gland. The aim of this study was to investigate the efficacy of FNAC of salivary gland lesions and to decide whether the radiologist could perform it or not. METHODS: We aspirated 162 salivary gland lesions of 56 patients undergoing biopsy and excision. A cytopathologist and a surgical pathologist made histopathological and cytological examinations in a blinded fashion. RESULTS: In the present study, among the FNAC performed in 162 patients with salivary gland masses, 15 (9%) were inadequate, and the remaining 147 were diagnostic. Of the 162 cases, 56 (35%) were also checked histologically. With FNAC there were 89 (55%) nonneoplastic and 58 (36%) neoplastic lesions. With FNAC 45 of the neoplastic lesions were benign and 13 malignant tumors. When cytohistopathological correlation was performed, the overall accuracy in diagnosing benign and malignant lesions was 91 and 78%, respectively, except for 2 malignant lymphoma cases. The sensitivity and specificity for benign and malignant lesions were 72 and 100%, respectively. The diagnostic sensitivity for all neoplastic and nonneoplastic lesions was 84% and the specificity 92%. There was 1 false-positive result, in which a nonneoplastic lesion was misdiagnosed as a Warthin's tumor. Fifteen (26%) cases were false negative, 12 of which were undefined. CONCLUSION: FNAC is a useful and reliable tool in the diagnosis of salivary gland masses when performed by a radiologist or a clinician. There are no contraindications, and complications are minimal. PMID- 17085954 TI - Surfactant protein A and D in human sinus mucosa: a preliminary report. AB - INTRODUCTION: Surfactant-associated proteins (SPs) play a crucial role in the innate defense system and serve as the initial step in the immune response to inhaled pathogens. SP-A and SP-D expression and function are altered in a variety of inflammatory and infectious diseases of the lungs, such as asthma, allergies, and cystic fibrosis, but their presence and function in the sinonasal cavity has not been investigated. The objective of this study was to test our hypothesis that SP-A and SP-D are present in the human sinus. MATERIALS AND METHODS: Sinus mucosal biopsies were performed in 8 patients undergoing endoscopic sinus surgery for chronic rhinosinusitis with nasal polyposis, pituitary tumors, and cerebrospinal fluid leak repairs. Expression of SP mRNA and protein by the sinus mucosa was detected by RT-PCR and immunoblot analysis, respectively. RESULTS: Analyses of mucosal biopsies from these patients revealed the presence of SP-A and SP-D mRNA and protein in all specimens. CONCLUSION: SP-A and SP-D are expressed in both normal and diseased human sinus tissue. Understanding the role of SPs in diseased and healthy states may elucidate their possible roles in innate immunity in the upper airway and allow us to develop novel treatments for sinonasal pathologies. PMID- 17085955 TI - Hodgkin's lymphoma: an appraisal of its biology and clinical manifestation. PMID- 17085956 TI - Pathobiology of classical Hodgkin lymphoma. AB - The World Health Organization has acknowledged the malignant nature of classical Hodgkin lymphoma (cHL), which encompasses four histological subtypes. The diagnosis of cHL is based on the detection of malignant Hodgkin and Reed Sternberg cells (HRSC) confirmed by immunophenotyping and the detection of growth patterns specific to each histological subtype. The pathologic HRSC arise from germinal center or immediate postgerminal cells that lack detectable immunoglobulin/B-cell antigen receptor expression, with a consequent loss of B cell identity; very few cHL cases are of T-cell origin. To escape apoptosis, which normally occurs in B cells with nonfunctioning antigen receptor machinery, HRSC develop concurrent antiapoptotic mechanisms by activation of nuclear factor kappaB or are rescued by Epstein-Barr virus infection. HRSC are characterized by a variable and inconstant immunophenotype, with a remarkable loss of lineage specific cell antigens and expression of antigens of other cell lineages. The master plan of B-cell identity in HRSC is disturbed not only at the immunoglobulin expression level, but also at the transcriptional factor level. HRSC are further characterized by profound cell cycle deregulation with futile replication, multinucleation and poly- and aneuploidy. Here, we review pathobiological aspects of cHL with respect to lymphomagenesis and routine diagnostics. PMID- 17085957 TI - Current strategies and new approaches in the treatment of Hodgkin's lymphoma. AB - As a result of continuous improvement in therapeutic options and their verification by large multicenter trials, Hodgkin's lymphoma (HL) has become one of the best curable cancers in adults. Nowadays, about 80-90% of patients in all stages achieve long-term survival. Nevertheless, these good results are threatened by treatment-associated toxicities such as infertility, cardiopulmonary toxicity and secondary malignancies. It is therefore the aim of future trial generations both to maintain excellent treatment results and to minimize late effects. At early stages, ongoing trials ask how many cycles of ABVD-like chemotherapy are necessary and if radiation doses might be further reduced or even omitted in favorable early-stage disease. In advanced stages, new combinations of chemotherapeutic drugs with higher dose densities are tested with or without the application of consolidating radiotherapy. The treatment of patients with relapsed HL depends on previous therapies with radiotherapy, chemotherapy or high-dose chemotherapy followed by autologous stem cell transplantation. For patients with multiple relapses, experimental treatment strategies include antibody- and small-molecule-based regimens. In this review we present current treatment strategies for patients with a first diagnosis of HL and relapsed HL as well as recent experimental therapeutic approaches. PMID- 17085958 TI - Biological inhibitory effects of the Chinese herb danggui on brain astrocytoma. AB - OBJECTIVE: Previous studies have demonstrated the utility of the traditional Chinese herb danggui in the treatment of chronic myelogenous leukemia. Our aim was to examine whether it might similarly be used to treat glioblastoma multiforme. METHODS: The lipid-soluble active ingredients of danggui were extracted with acetone (AS-AC) or chlorophenol (AS-CH) and their antiproliferative and proapoptotic effects were studiedin vitro on cultured GBM 8401 cells and in vivoon tumors in nude mice. RESULTS: After a 24-hour treatment, either AS-AC or AS-CH at a lower (50 micro g/ml) and a higher concentration (100 micro g/ml) significantly inhibited the proliferative activity of GBM 8401 cultured cells by 30-50%, as well as the expression of cathepsin B and vascular endothelial growth factor (VEGF). In nude mice, the growth of the tumor was inhibited by 30% by AS-CH or AS-AC (20 mg/kg; p < 0.05) and by 60% by AS-CH or AS AC (60 mg/kg; p < 0.05). AS-AC and AS-CH also significantly inhibited microvessel formation in the tumors of nude mice. CONCLUSIONS: Danggui may inhibit tumor growth by reducing the level of VEGF and the proapoptotic protein, cathepsin B. Thus, danggui may be useful in the treatment of high-grade astrocytomas. PMID- 17085959 TI - Vascular wall maturation and prolonged angiogenic effect by endothelial-specific platelet-derived growth factor expression. AB - BACKGROUND: The implementation of angiogenic gene therapy at clinics is hindered by the transience of the therapeutic effect. Recruiting vascular wall smooth muscle cells, a process termed 'maturation', can stabilize newly formed vessels. OBJECTIVE: To induce angiogenesis followed by vessel maturation in a murine ischemic limb model by endothelial cell-specific promoter regulated expression of vascular endothelial growth factor (VEGF) and platelet-derived growth factor-BB (PDGF-BB). METHODS: We constructed adenoviral vectors containing angiogenic factors VEGF and PDGF-B regulated by a modified preproendothelin-1 (PPE-1-3x) promoter and investigated their angiogenic effect in a murine ischemic limb model. RESULTS: VEGF gene therapy increased perfusion and the vessel density in the limb shortly after expression with PPE-1-3x promoter or cytomegalovirus (CMV) promoter vectors, but only PPE-1-3xVEGF treatment exhibited a sustained effect. Expression of PDGF-B by PPE-1-3x promoter resulted in morphological maturation of the vasculature and further increased the perfusion, while nonspecific expression of PDGF-B with CMV promoter had no therapeutic effect. Regulation of dual therapy with VEGF and PDGF-B by PPE-1-3x promoter resulted in an early-onset, sustained angiogenic effect accompanied by vessel maturation. CONCLUSIONS: Systemic gene therapy with the angiogenic factors VEGF and PDGF-B under angiogenic- endothelial cell-specific regulation was effective in inducing functionally and morphologically mature vasculature. PMID- 17085960 TI - Delusional denial of pregnancy as a special form of Cotard's syndrome: case report and review of the literature. AB - The following case report describes a pregnant woman who was convinced that her pregnancy was non-existent despite being in an advanced stage of clinically obvious pregnancy. The nosologically unspecific syndrome of denial of pregnancy is discussed by reviewing the literature. Based on the existing literature it will be explained why this specific syndrome is considered to indicate a special form of Cotard's syndrome. PMID- 17085961 TI - Postpartum psychoses over five decades. PMID- 17085962 TI - Translation and transcription: the dual functionality of LysRS in mast cells. AB - In the post genome project era, it is well established that the human genome contains a smaller number of genes than expected. The complexity found in higher organisms can be explained if proteins are multifunctional. Indeed, recent studies are continuing to reveal proteins that are capable of a broad repertoire of functions. A good paradigm for multifunctionality can be found in the amino acyl tRNA synthetases (aaRSs), an ancient conserved family of proteins. This unique family, which is comprised of 20 different enzymes, is well known for its participation in protein synthesis. Several studies have described numerous examples of these "housekeeping" proteins taking part in extensive critical cellular activities. In this review, we focus on a member of that family, lysyl tRNA synthetase (LysRS), which has been shown to have a dual functionality. In addition to its contribution to the translation process, LysRS also takes part in the regulation of MITF and USF2 target genes. This phenomenon was first described in mast cells. PMID- 17085963 TI - Jab1 as a mediator of nuclear export and cytoplasmic degradation of p53. AB - Jun activation domain-binding protein 1 (Jab1) is involved in various cellular mechanisms including development in Drosophila and mouse, cell cycle control and signal transduction pathways. Recent studies also determined that Jab1 functions as a nuclear exporter and inducer of cytoplasmic degradation for several proteins including p53, p27, capsid of West Nile virus, and Smad4/7 proteins. In particular, p53 is shown to bind to and to be exported into the cytoplasm by Jab1, which helps to maintain low levels of p53 under normal conditions. This review was undertaken in an effort to understand the biological significance of the homeostasis of p53 as maintained in the presence of Jab1. Based on our observations, we have provided potential mechanistic hypotheses for the nuclear export of p53 in coordination with Jab1 and the role of other factors in these processes. PMID- 17085964 TI - Mouse transthyretin-related protein is a hydrolase which degrades 5 hydroxyisourate, the end product of the uricase reaction. AB - Uric acid is the end product of the purine degradation pathway in humans. It is catabolized to allantoin by urate oxidase or uricase (E.C. 1.7.3.3.) in most vertebrates except humans, some primates, birds, and certain species of reptiles. Here we provide evidence that mouse transthyretin-related protein facilitates the hydrolysis of 5-hydroxyisourate, the end product of the uricase reaction. Mutagenesis experiments showed that the residues that are absolutely conserved across the TRP family, including His11, Arg51, His102, and the C-terminal Tyr-Arg Gly-Ser, may constitute the active site of mTRP. Based on these results, we propose that the transthyretin-related proteins present in diverse organisms are not functionally related to transthyretin but actually function as hydroxyisourate hydrolases. PMID- 17085965 TI - Function of rax2p in the polarized growth of fission yeast. AB - Cell polarity is critical for the division, differentiation, migration, and signaling of eukaryotic cells. RAX2 of budding yeast encodes a membrane protein localized at the cell cortex that helps maintain the polarity of the bipolar pattern. Here, we designate SPAC6f6.06c as rax2+ of Schizosaccharomyces pombe, based on its sequence homology with RAX2, and examine its function in cell polarity. S. pombe rax2+ is not essential, but Deltarax2 cells are slightly smaller and grow slower than wild type cells. During vegetative growth or arrest at G1 by mutation of cdc10, deletion of rax2+ increases the number of cells failing old end growth just after division. In addition, this failure of old end growth is dramatically increased in Deltatea1Deltarax2, pointing to genetic interaction of rax2+ with tea1+. Deltarax2 cells contain normal actin and microtubule cytoskeletons, but lack actin cables, and the polarity factor for3p is not properly localized at the growing tip. In Deltarax2 cells, and endogenous rax2p is localized at the cell cortex of growing cell tips in an actin- and microtubule-dependent manner. However, Deltarax2 cells show no defects in cell polarity during shmoo formation and conjugation. Taken together, these observations suggest that rax2p controls the cell polarity of fission yeast during vegetative growth by regulating for3p localization. PMID- 17085966 TI - Angucyclines Sch 47554 and Sch 47555 from Streptomyces sp. SCC-2136: cloning, sequencing, and characterization. AB - The entire gene cluster involved in the biosynthesis of angucyclines Sch 47554 and Sch 47555 was cloned, sequenced, and characterized. Analysis of the nucleotide sequence of genomic DNA spanning 77.5-kb revealed a total of 55 open reading frames, and the deduced products exhibited strong sequence similarities to type II polyketide synthases, deoxysugar biosynthetic enzymes, and a variety of accessory enzymes. The involvement of this gene cluster in the pathway of Sch 47554 and Sch 47555 was confirmed by genetic inactivation of the aromatase, including a portion of the ketoreductase, which was disrupted by inserting the thiostrepton gene. PMID- 17085967 TI - Histone deacetylation is involved in activation of CXCL10 upon IFNgamma stimulation. AB - Histone deacetylase (HDAC) activity is commonly associated with transcriptional repression. However, there is also evidence for a function in transcriptional activation. Previous studies have demonstrated a fundamental role of deacetylase activity in IFNa-responsive gene transcription. In the case of type II IFN (IFNgamma) results are controversial: some genes require HDAC activity, while transcription of others is repressed by HDAC. To investigate the effect of HDAC on transcription of an IFNgamma-activated gene, real-time PCR was used to measure CXCL10 mRNA in Hela cells stimulated with IFNgamma in the presence or absence of the HDAC inhibitor TSA. Chromatin imunoprecipitation combined with real-time PCR was used to check acetylation of histone H4 and recruitment of the STAT1 complex to the ISRE locus of the CXCL10 gene. Activation of CXCL10 transcription in response to IFNgamma was paralleled by a decrease in histone H4 acetylation and an increase in recruitment of the STAT1 complex to the CXCL10 ISRE locus. The transcription of CXCL10 and histone H4 deacetylation were blocked by TSA, but the latter had no obvious affect on recruitment of the STAT1 complex. Our data indicate that IFNgamma and STAT-dependent gene transcription requires the participation of HDAC, as does the IFNalpha-STAT pathway. PMID- 17085968 TI - Altered gene expression of caspase-10, death receptor-3 and IGFBP-3 in preeclamptic placentas. AB - Enhanced apoptosis has been observed in the placentas of women with preeclampsia, but few studies have examined changes at the molecular level. This study was designed to detect genes specifically expressed in full-term preeclamptic placentas. Tissue samples were collected immediately after cesarean delivery from 11 normal and 8 preeclamptic placentas at 35-40 weeks of gestation. Total RNAs were extracted and hybridized to a cDNA microarray. Results were confirmed by reverse-transcription polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry. Hematoxylin and eosin and TUNEL staining were also performed to confirm apoptosis in preeclamptic placentas. Among 205 genes, three were up- or down-regulated in preeclamptic placentas. The expression of caspase 10 and death receptor 3 (DR-3) was significantly increased, whereas insulin-like growth factor binding protein-3 (IGFBP-3) was strongly down-regulated. RT-PCR analysis and Western blotting confirmed these effects. Immunohistochemical analysis showed that the DR-3, caspase-10 and IGFBP-3 proteins were localized in the syncytial membrane. Apoptosis in the trophoblast was also increased in term placentas from women with pregnancies complicated by preeclampsia. These results suggest that caspase-10, DR-3 and IGFBP-3 are involved in apoptosis in the preeclamptic placenta. PMID- 17085969 TI - Induction of immunity against hepatitis B virus surface antigen by intranasal DNA vaccination using a cationic emulsion as a mucosal gene carrier. AB - Delivery of DNA vaccines to airway mucosa would be an ideal method for mucosal immunization. However, there have been few reports of a suitable gene delivery system. In this study we used a cationic emulsion to immunize mice via the intranasal route with pCMV-S coding for Hepatitis B virus surface antigen (HBsAg). Complexing pCMV-S with a cationic emulsion dramatically enhanced HBsAg expression in both nasal tissue and lung, and was associated with increases in the levels of HBs-specific Abs in serum and mucosal fluids, of cytotoxic T lymphocytes (CTL) in the spleen and cervical and iliac lymph nodes, and of delayed-type hypersensitivity (DTH) against HBsAg. In contrast, very weak humoral and cellular immunities were observed following immunization with naked DNA. In support of these observations, a higher proliferative response of spleenocytes was detected in the group immunized with the emulsion/pCMV-S complex than in the group immunized with naked pCMV-S. These findings may facilitate development of an emulsion-mediated gene vaccination technique for use against intracellular pathogens that invade mucosal surfaces. PMID- 17085970 TI - Methylation of the mouse DIx5 and Osx gene promoters regulates cell type-specific gene expression. AB - Dlx5 and Osx are master regulatory proteins essential for initiating the cascade leading to osteoblast differentiation in mammals, but the mechanism of osteoblast specific expression is not fully understood. DNA methylation at CpG sequences is involved in tissue and cell type-specific gene expression. We investigated the methylation status of Dlx5 and Osx in osteogenic and nonosteogenic cell lines by methylation-specific PCR (MSP). The CpG dinucleotides of the Dlx5 and Osx promoter regions were unmethylated in osteogenic cell lines transcribing these genes but methylated in nonosteogenic cell lines. Treatment of C2C12 cells with 5 AzadC induced dose- and time-dependent expression of Dlx5 and Osx mRNA by demethylating the corresponding promoters. Furthermore the mRNAs for the osteoblast markers ALP and OC, which were undetectable in untreated cells, gradually increased after 5-AzadC treatment. In addition, BMP-2 stimulation induced Dlx5 expression by hypomethylating its promoter. These findings suggest that DNA methylation plays an important role in cell type-specific expression of Dlx5 and Osx. PMID- 17085971 TI - Changes in reproductive function and white blood cell proliferation induced in mice by injection of a prolactin-expressing plasmid into muscle. AB - Prolactin (PRL) is a pituitary hormone involved in various physiological processes, including lactation, mammary development, and immune function. To further investigate the in vivo and comparative endocrine roles of PRL, mouse PRL cDNA fused to the cytomegalovirus promoter, was introduced into muscle by direct injection. Previously we studied the function of rat PRL using the same protocol. PRL mRNA was detected in the muscle following injection by RT-PCR and subsequent Southern blot analysis. PRL was also detected and Western blot analysis revealed a relatively high level of serum PRL. In the pCMV-mPRL-injected female mice, the estrous cycle was extended, especially in diestrus stage and the uterus thickening that was shown in normal estrous stage was not observed. In the pCMV mPRL-injected male mice, new blood vessels were first found at 5 weeks of age and fully developed blood vessels were found after 8 weeks in the testis. The number of Leydig cells increased within the testis and the testosterone level in serum was observed high. Finally, the number of white blood cells (WBCs) increased in the pCMV-mPRL-injected mice. The augmentation of WBCs persisted for at least 20 days after injection. When injection was combined with adrenalectomy, there was an even greater increase in number of WBCs, especially lymphocytes. This increase was returned normal by treatment with dexamethansone. Taken together, our data reveal that intramuscularly expressed mouse PRL influences reproductive functions in female, induces formation of new blood vessels in the testis, and augments WBC numbers. Of notice is that the Leydig cell proliferation with increased testosterone was conspicuously observed in the pCMV-mPRL-injected mice. These results also suggest subtle difference in function of PRL between mouse and rat species. PMID- 17085972 TI - Establishment of an assay for P2X7 receptor-mediated cell death. AB - The P2X7 receptor, an ATP-gated cation channel, induces cell death in immune cells and is involved in neurodegenerative diseases. Although the receptor plays various roles in these diseases, the cellular mechanisms involved are poorly understood and antagonists are limited. Here, the development of a cell-based assay for human P2X7 receptor is reported. We established permanent lines of HEK 293 cells expressing a high level of hP2X7 receptor. Functional activity of the hP2X7 receptor was confirmed by whole-cell patch recording of ATP-induced ion currents. Prolonged exposure to ATP resulted in death of the hP2X7-expressing HEK 293 cells and this cell death could be quantified. Two known P2X7 antagonists, PPADS and KN-62, blocked ATP-induced death in a concentration-dependent manner. Thus, this assay can be used to screen for new antagonists of hP2X7 receptors. PMID- 17085973 TI - TATA-binding protein-related factor 2 is localized in the cytoplasm of mammalian cells and much of it migrates to the nucleus in response to genotoxic agents. AB - TBP (TATA-binding protein)-related factor 2 (TRF2) regulates transcription during a nuber of cellular processes. We previously demonstrated that it is localized in the cytoplasm and is translocated to the nucleus by DNA-damaging agents. However, the cytoplasmic localization of TRF2 is controversial. In this study, we reconfirmed its cytoplasmic localization in various ways and examined its nuclear migration. Stresses such as heat shock, redox agents, heavy metals, and osmotic shock did not affect localization whereas genotoxins such as methyl methanesulfonate (MMS), cisplatin, etoposide, and hydroxyurea caused it to migrate to the nucleus. Adriamycin, mitomycin C and gamma-rays had no obvious effect. We determined optimal conditions for the nuclear migration. The proportions of cells with nuclei enriched for TRF2 were 25-60% and 5-10% for stressed cells and control cells, respectively. Nuclear translocation was observed after 1 h, 4 h and 12 h for cisplatin, etoposide and MMS and hydroxyurea, respectively. The association of TRF2 with the chromatin and promoter region of the proliferating cell nuclear antigen (PCNA) gene, a putative target of TRF2, was increased by MMS treatment. Thus TRF2 may be involved in genotoxin-induced transcriptional regulation. PMID- 17085974 TI - A WD40 repeat protein, Arabidopsis Sec13 homolog 1, may play a role in vacuolar trafficking by controlling the membrane association of AtDRP2A. AB - Dynamin-related protein 2A (AtDRP2A, formally ADL6), a member of the dynamin family, is critical for protein trafficking from the TGN to the central vacuole. However, the mechanism controlling its activity is not well understood in plant cells. We isolated Arabidopsis sec13 homolog1 (AtSeh1) that interacts with AtDRP2A by a yeast two-hybrid screening. AtSeh1 has four WD40 motifs and amino acid sequence homology to Sec13, a component of COPII vesicles. Coimmunoprecipitation and protein pull-down experiments demonstrated specific interaction between AtSeh1 and AtDRP2A. AtSeh1 bound to the pleckstrin homology domain of AtDRP2A in competition with the C-terminal domain of the latter, and this resulted in inhibition of the interaction between AtDRP2A and PtdIns3P in vitro. AtSeh1 localized to multiple locations: the nucleus, the prevacuolar compartment and the Golgi complex. Based on these results we propose that AtSeh1 plays a role in regulating cycling of AtDRP2A between membrane-bound and soluble forms. PMID- 17085975 TI - Oxidative modification of cytochrome c by hydrogen peroxide. AB - Oxidative alteration of mitochondrial cytochrome c has been linked to disease and is one of the causes of pro-apoptotic events. We have investigated the modification of cytochrome c by H2O2. When cytochrome c was incubated with H2O2, oligomerization of the protein increased and the formation of carbonyl derivatives and dityrosine was stimulated. Radical scavengers prevented these effects suggesting that free radicals are implicated in the H2O2-mediated oligomerization. Oligomerization was significantly inhibited by the iron chelator, deferoxamine. During incubation of deoxyribose with cytochrome c and H2O2, damage to the deoxyribose occurred in parallel with the release of iron from cytochrome c. When cytochrome c that had been exposed to H2O2 was analyzed by amino acid analysis, the tyrosine, histidine and methionine residues proved to be particularly sensitive. These results suggest that H2O2-mediated cytochrome c oligomerization is due to oxidative damage resulting from free radicals generated by a combination of the peroxidase activity of cytochrome c and the Fenton reaction of free iron released from the oxidatively-damaged protein. PMID- 17085976 TI - Ovarian tumors in Rbp9 mutants of Drosophila induce an immune response. AB - The Drosophila protein, Rbp9, is homologous to human Hu, which is reported to be involved in small cell lung cancer. Rbp9 functions in cystocyte differentiation, and mutations in Rbp9 cause ovarian tumors. Here we show that the antimicrobial peptide, Attacin, is upregulated in Rbp9 mutants, especially in ovaries where tumors form. Upregulation seems to result from activation of the NF-kappaB pathway since we detected nuclear localization of Relish in Rbp9 mutant ovaries but not in wild type ovaries. Inactivation of NF-kappaB in the Rbp9 mutant allows prolonged survival of malformed egg chambers. We conclude that Drosophila initiates an anti-tumor defense response via activation of NF-kappaB. PMID- 17085977 TI - Induction of a salicylic acid glucosyltransferase, AtSGT1, is an early disease response in Arabidopsis thaliana. AB - Endogenous salicylic acid (SA) and its predominant conjugates, SA 2-O-beta-D glucoside (SAG) and the glucose ester of SA (SGE), increase dramatically during plant defense responses. Here I report the isolation and characterization of an Arabidopsis thaliana UDP-glucose:SA glucosyltransferase1 (AtSGT1) gene using a tobacco SGT gene previously reported, whose product catalyzes the formation of both SAG and SGE. The recombinant AtSGT1 protein had significant activities with SA and benzoic acid, and synthesized SAG and SGE. Northern blot analysis showed that AtSGT1 was rapidly induced both by exogenous SA and infection with the bacterial pathogen Pseudomonas syringae, indicating that pathogen-inducible AtSGT1 expression is an early disease response and may be involved in the accumulation of glucosyl SA during pathogenesis. PMID- 17085978 TI - Satellite cells isolated from adult Hanwoo muscle can proliferate and differentiate into myoblasts and adipose-like cells. AB - This study examined whether adult bovine muscle satellite cells from 30-month-old Hanwoo cattle are multipotential. The satellite cells were found to have the potential to proliferate and differentiate into myoblasts with the formation of multinucleated cells. In addition, treatment with the peroxisome proliferator activating receptor-gamma (PPARgamma) agonist, rosiglitazone, promoted their trans-differentiation into adipocytes with significant increases in glycerol accumulation and glycerol-3-phosphate dehydrogenase activity. Western blot analysis revealed that increased levels of the adipocyte fatty acid-binding protein, PPARgamma and of CCAAT/enhancer-binding protein were closely related to rosiglitazone-induced differentiation of the cells. These findings demonstrate that satellite cells from adult Hanwoo cattle are multipotent, and that their trans-differentiation into adipocytes can be induced by rosiglitazone. PMID- 17085979 TI - Women with chest pain and normal coronary angiograms: no longer a benign syndrome? PMID- 17085980 TI - Achievement of national cholesterol education program goals by patients with dyslipidemia in rural ambulatory care settings. AB - The Third Adult Treatment Panel (ATP III) of the National Cholesterol Education Program provides guidelines for managing dyslipidemia; however, studies from large centers find that most dyslipidemic patients fail to achieve management goals. Few data exist on lipid management in rural settings. To determine the proportion of rural dyslipidemic patients achieving ATP III goals, records of 461 patients were reviewed from 4 practices. Only 54% of the patients with dyslipidemia achieved ATP III goals. Patients with diabetes or with a family history of premature coronary heart disease were less likely to achieve ATP III goals (odds ratio 0.56; 95% confidence interval, 0.38-0.84 and odds ratio 0.42; 95% confidence interval, 0.25-0.71, respectively). Patients taking statins were more likely to achieve goals (odds ratio 3.23; 95% confidence interval, 2.13 4.89). These results indicate that a significant proportion of patients with dyslipidemia in rural practices do not achieve management goals. Strategies to improve lipid management in rural practices are needed. PMID- 17085981 TI - Coronary calcification in body builders using anabolic steroids. AB - The authors measured coronary artery calcification as a means of examining the impact of anabolic steroids on the development of atherosclerotic disease in body builders using anabolic steroids over an extended period of time. Fourteen male professional body builders with no history of cardiovascular disease were evaluated for coronary artery calcium, serum lipids, left ventricular function, and exercise-induced myocardial ischemia. Seven subjects had coronary artery calcium, with a much higher than expected mean score of 98. Six of the 7 calcium scores were >90th percentile. Mean total cholesterol was 192 mg/dL, while mean high-density lipoprotein was 23 mg/dL and the mean ratio of total cholesterol to high-density lipoprotein was 8.3. Left ventricular ejection fraction ranged between 49% and 68%, with a mean of 59%. No subject had evidence of myocardial ischemia. This small group of professional body builders with a long history of steroid abuse had high levels of coronary artery calcium for age. The authors conclude that in this small pilot study there is an association between early coronary artery calcium and long-term steroid abuse. Large-scale studies are warranted to further explore this association. PMID- 17085982 TI - Pharmacist-led cardiac risk reduction model. AB - The Cardiovascular Risk Reduction Clinic (CRRC) is a pharmacist-coordinated care model that incorporates the 4 major modifiable risk factors into a single treatment program. The authors aimed to assess the effectiveness of the CRRC model by comparing the Framingham point score before and after CRRC intervention. Retrospective data between 2001 and 2002 were analyzed for 375 veterans (age 65+/ 10 years) with diabetes (88.8%) or coronary artery disease (44%) referred to the CRRC for intensive cardiac risk management. Total Framingham point score decreased from 14.5 at baseline to 13.6 after CRRC intervention (mean change 0.9+/-0.3). When considering only the patients not at target goals at baseline (n=200), significant improvements in guideline adherence was observed in low density lipoprotein cholesterol, systolic blood pressure, hemoglobin A1c, and smoking. The CRRC model reduces the risk of cardiovascular events as assessed by Framingham point score in patients with established coronary artery disease and/or diabetes. PMID- 17085983 TI - The acute effect of aerobic exercise on brachial artery endothelial function in renal transplant recipients. AB - This study compared the effect of a 30-minute walk on brachial artery endothelial vasodilatation in kidney transplant (KT) recipients and healthy controls (HCs). Endothelial-dependent vasodilatation was measured by ultrasound before and after exercise. The HCs experienced a significant increase in vasodilatation after exercise 1 minute postocclusion when compared with the KT recipients (22%+/-13% vs 3%+/-4%; P<.05). Also, the HCs had a significantly higher vasodilatation from pre-treadmill walk to post-treadmill walk (1 minute postocclusion) when compared with KT recipients (from 3%+/-6% to 22%+/-13% vs 1%+/-3% to 3%+/-4%; P<.05). This acute vasodilatory response observed in the HCs may be related to the immediate release of nitric oxide and the combined response to shear stress and exercise. The KT recipients had several coronary artery disease risk factors that may have adversely affected endothelial function. PMID- 17085984 TI - Gender differences in adhering to national guidelines in a community lipid clinic. AB - The aggressive treatment of hypercholesterolemia improves morbidity and mortality in patients with a history of cardiovascular disease irrespective of gender. Electronically tracked data on 4324 patients enrolled in a community lipid clinic were analyzed for gender differences in lipid values and adherence to national guidelines in lipid management. Women were older, more likely to be diabetic and/or hypertensive, and have a family history of coronary heart disease. Women also had higher initial total cholesterol, low-density lipoprotein, and triglyceride levels and were more likely to be at goal at entry for high-density lipoprotein. Men were more likely to have coronary heart disease and lower high density lipoprotein and were more likely to be at goal at entry for triglyceride level. There were no gender differences in low-density lipoprotein at goal at entry or rate of current smoking. All lipid parameters appeared better with age, irrespective of gender. More men than women reached goal for their low-density lipoprotein (61.5% vs 51.7%) and triglyceride (36.9% vs 25.1%) levels, whereas more women than men reached goal for their high-density lipoprotein (33.1% vs 22.2%). The authors conclude that there are significant gender differences in lipid values at entry to a community lipid clinic and in achieving National Cholesterol Education Program targets following the initiation of therapy. A possible gender-independent survivorship effect exists for low-density lipoprotein and high-density lipoprotein. PMID- 17085985 TI - Lower is better--the contemporary concept of low-density lipoprotein lowering in the preventive management of cardiovascular risk: does this apply to all patients? AB - The number of multiple outcomes-based clinical trials evaluating the use of statin drugs for lowering cardiovascular risk continues to grow, incorporating patients requiring primary and secondary preventive care. This review surveys the most recently published studies and identifies both the more extensive patient population that may benefit from primary preventive care and the concept of aggressive titration of the statin drug to improve the prognosis of the patient undergoing secondary preventive care. Data on the elderly patient, the female patient, and the diabetic patient are reviewed, as are the possible mechanisms of action of the statins in modifying cardiovascular risk. The pleiotropic effects and anti-inflammatory capabilities of the drugs are also reviewed. Conclusions are drawn regarding the contemporary use of statins in the primary and secondary preventive management of patients to significantly reduce cardiovascular risk. PMID- 17085986 TI - Acute myocardial infarction and acute coronary syndrome: then and now (1950 2005). AB - Advances in the prevention, diagnosis, and treatment of acute myocardial infarction (AMI) and acute coronary syndrome (ACS) have been remarkable since the mid-20th century. Even the clinical terminology used to describe some of the various components of ACS have undergone change, while the latter term itself represents a fairly recent addition to the medical lexicon. Although there have been dramatic changes in the diagnostic and therapeutic interventions used and impressive declines in morbidity and mortality, the differential diagnosis and complications of AMI and ACS remain as challenging now as they were a half century ago. This article presents in detail the medical understanding of AMI in the mid-20th century and how physicians of that era managed it and its complications, and contrasts this with current evidence-based knowledge and interventions. PMID- 17085987 TI - CHARISMA and TROPHY. PMID- 17085988 TI - Updated secondary prevention guidelines for atherosclerotic disease. PMID- 17085989 TI - The girth of a nation--a call to arms...and legs. PMID- 17085990 TI - Dermatology is a specialty; dermatology is not a subspecialty. PMID- 17085991 TI - The many faces of molluscum contagiosum: the fool factor. PMID- 17085992 TI - Top 10 head lice concerns. PMID- 17085993 TI - Phase 2 trial of the continuous IV administration of interferon-beta in patients with disseminated malignant melanoma. AB - BACKGROUND: Interferons have been reported to significantly contribute to tumor suppression via both induction of p53 gene expression and inhibition of angiogenesis. OBJECTIVE: The assessment of treatment toxicity and antitumoral effectiveness of continuous IV administration of interferon-beta based on an overall evaluation of laboratory, radiographic, and clinical parameters observed during the trial. METHODS: The authors treated patients with advanced malignant melanoma with continuous IV infusions of 1 x 10(6) IU interferon-beta daily ( approximately 0.6 x 10(6) IU interferon-beta/m2 daily). RESULTS: Continuous IV administration of interferon-beta had no significant effect on overall patient outcome. Interferon side effects were not a reason for treatment discontinuation in any of the patients observed during this trial. CONCLUSIONS: Continuous IV interferon-beta had no significant effect on overall patient outcome in a group of patients with advanced malignant melanoma. To our knowledge, this is the first report on the continuous IV administration of interferon-beta in patients with advanced malignant melanoma. PMID- 17085994 TI - Psoriasis and pentoxifylline: a clinical, histopathologic, and immunohistochemical evaluation. AB - OBJECTIVE: The aim of this study was to examine the clinical and histopathologic effects of pentoxifylline on psoriasis, compared with placebo. METHODS: Sixty-one outpatients with active psoriasis were randomly assigned to either of 2 groups: one was given pentoxifylline 400 mg tid PO, and the other, placebo. Fifty-six patients concluded the study. They were evaluated clinically and by laboratory parameters before and after 8 weeks of treatment. Pretreatment and posttreatment biopsies were taken. Initial sections were stained with hematoxylin-eosin. Further cuts were immunostained for cytokeratins 10, 14, and 16. RESULTS: Clinical and histologic improvement did not show statistically significant differences between the groups. No laboratory abnormalities or serious reactions related to the drug were observed. CONCLUSIONS: No statistical difference was seen when the treatment group was compared with the control group. Pentoxifylline is a well tolerated and safe drug, but its efficacy in psoriasis appears to be limited. PMID- 17085995 TI - A treatment strategy for psoriasis: transitioning from systemic therapy to biologic agents. AB - Systemic agents such as methotrexate and cyclosporine are commonly used in the treatment of psoriasis. Long-term continuous use is not recommended due to potential organ toxicity, myelosuppression, and carcinogenicity. Abrupt cessation of systemic agents without tapering can lead to flare-up and rebound of psoriasis. The addition of a biologic agent during transitional therapy from a systemic agent is a good strategic maneuver to prevent a potential rebound complication. Even with a gradual cessation of systemic agents, the psoriasis will eventually relapse if biologic agents are not added to the treatment regimen. No additional toxicities or adverse events are evident during combination therapy with a systemic agent and a biologic agent. PMID- 17085996 TI - Solodyn (Minocycline HCl, USP) extended-release tablets. PMID- 17085997 TI - Hyaluronic acid for restoring earlobe volume. AB - Cosmetic disfigurement of the earlobe, including laxity and sagging skin, comes with aging. Restoring the earlobe volume is a procedure indicated for patients seeking a youthful facial appearance. Hyaluronic acid is a good choice for this procedure. Most fillers on the market today are supplied with a small needle, yet a larger-gauge needle allows the introduction of the product under lower pressure, making it easier and faster to fill tight areas. This article describes a useful approach to enhance the hyaluronic acid injection, resulting in a fast and effective method to restore earlobe volume. PMID- 17085998 TI - Cupping. PMID- 17085999 TI - Bite the hand that sprays you. AB - A 56-year-old man presented with a cutaneous lesion on his right hand (Figure 1). Approximately 6 weeks previously, he had traveled to Costa Rica for missionary work. During his travel he used a diethyltoluamide-containing insect repellant, but spent one night in the jungle without a mosquito net. Four weeks after his return, he noted a 3 x 2.5-cm ulcer with raised edges and surrounding erythema on the dorsum of his right hand. The patient recalled applying insect repellant with his right hand to other exposed areas of his body but was remiss in applying it to the right hand itself. On examination, the patient was noted to have an ulcerated nodule near his right earlobe (Figure 2) and a second 2 x 1-cm ulcer on the volar aspect of his right wrist (Figure 3). There were no mucocutaneous lesions noted. During the next several weeks, numerous nodules developed in a sporotrichoid pattern on the extensor surface of his right arm (Figure 4). Skin biopsy was performed at the time of initial evaluation and revealed cutaneous leishmaniasis due to Leishmania panamensis. After discussing the different treatment options, miltefosine was administered orally for 28 days. The patient experienced an excellent response to therapy. PMID- 17086000 TI - Telangiectasia macularis eruptiva perstans: unusual presentation and treatment. AB - A 41-year-old woman presented with a 2-month history of pruritus and a generalized dermatitis that developed initially on the head and spread to the trunk, legs, and buttocks. The pruritus caused extreme discomfort and was not relieved by antihistamines or topical steroid treatment. The patient denied flushing, syncope, and vomiting. Her medical history included asthma treated with salmeterol/fluticasone propionate inhaler, and status post silicone breast augmentation. Physical examination revealed a papular dermatitis on the trunk and extremities composed of lesions up to 0.5 cm in diameter, surrounded by excoriation marks (Figure 1). There was no hepatosplenomegaly or lymphadenopathy. Darier's sign was negative. Results of complete blood count, peripheral blood film examination, and liver function tests were all with normal limits. A biopsy specimen taken from a lesion and stained with hematoxylin-eosin showed telangiectasias, with an increased number of mast cells around blood vessels (Figure 2). Positive Giemsa (Figure 3) and c-kit stain (Figure 4) indicated an increased number of mast cells. Bone marrow aspiration and total body CT performed to rule out systemic involvement showed no pathology. Protein electrophoresis was normal. Serum tryptase and histamine were within normal limits, and 24-hour urine collection for histamine was normal. Narrow-band UV-B treatment was begun 3 times weekly, reduced to twice weekly after 2 months, and then stopped. The first few treatments resulted in significant relief of the pruritus and regression of lesions. After 3 months without treatment, the patient remained free of pruritus and lesions. PMID- 17086001 TI - Perinuclear antineutrophil cytoplasmic antibody-associated vasculitis in a patient with Graves' disease treated with methimazole. AB - A 15- year-old Caucasian girl presented with a 1-week history of necrotic ulcers on her bilateral lower extremities, associated with fever and malaise. She had had similar ulcerations on and off over the past 3 years. She had a medical history of Graves' disease, diagnosed in 1999, for which she was taking methimazole (MMI). On physical examination, there were multiple necrotic ulcers with elevated, inflamed borders and an overlying eschar on the bilateral distal extremities (Figure 1). There was one hemorrhagic, flaccid bulla on the lateral aspect of the ankle. Her complete blood count, comprehensive metabolic panel, prothrombin time, and international normalized ratio were within normal limits. Histopathology revealed a neutrophilic vasculitis involving small as well as medium-sized blood vessels. The overlying epidermis and upper dermis showed necrosis (Figure 2). Antinuclear antibody (ANA) was positive at 1:160. Perinuclear (p-) antineutrophil cytoplasmic antibody (ANCA) was positive at 1:320. Cytoplasmic ANCA was negative. The patient did not meet any clinical criteria for a diagnosis of lupus erythematosus. Direct immunofluorescence revealed IgM, complement C3, and fibrin in small- and medium-caliber blood vessels in the dermis, consistent with a leukocytoclastic vasculitis of small- and medium-caliber blood vessels (Figure 3). MMI was stopped and the patient was given prednisone, tapered over 5 days. Her thyroid function remained stable, and she did not develop new necrotic ulcers. One month later, the patient developed urticarial lesions on her lower extremities. Histopathology revealed a lymphocytic vascular reaction. No eosinophils or neutrophils were seen. These newer urticarial lesions resolved. The patient underwent debridement of the thick, necrotic ulcerative eschars. To date, her lesions have resolved (Figure 4). An ANA drawn 5 months after withdrawal of MMI was negative, and p-ANCA was 1:67. PMID- 17086002 TI - Aggressive squamous cell carcinoma in Kindler syndrome. AB - A 57-year-old Hispanic man with a personal and family history of bullae and photosensitivity presented with a fungating, ulcerated squamous cell carcinoma on his left hand (Figure 1). Physical examination showed conjunctival injection, ectropion, symblepharon, urethral stricture, loss of teeth, short stature, and nail dystrophy. There was reticulated erythema, atrophy, hyperpigmentation and hypopigmentation, and telangiectasia of sun-exposed skin of the face, neck, and hands consistent with poikiloderma (Figure 2). In addition, there was foreshortening of the left thumb and sclerodermoid changes of his hands (Figure 3). Radiation therapy was applied to shrink the tumor before a local excision was performed. However, a local recurrence followed and axillary lymph nodes became clinically palpable, necessitating amputation and lymph node dissection. Extensive histologic evaluation of the specimen obtained following left arm amputation and lymph node dissection showed moderate-to-poorly differentiated deeply invasive squamous cell carcinoma. Two of 3 axillary lymph nodes were positive for metastatic carcinoma. A random biopsy of the trunk showed epidermal atrophy, telangiectasia, a perivascular lymphocytic infiltration, and pigment laden macrophages consistent with poikiloderma. Electron microscopy illustrated extensive reduplication of the basement membrane, with loops, curls, and free extensions of the basal lamina in the superficial dermis; reduced numbers of hemidesmosomes and anchoring fibrils; and a basement membrane focally devoid of basal cells (Figure 4). On the basis of the clinical features and the characteristic basement zone changes, a diagnosis of Kindler syndrome was made. PMID- 17086003 TI - Botulinum toxin and pregnancy. AB - The overall use of botulinum toxin type A in cosmetic procedures continues to rise, and the incidental exposures of pregnant women to the drug can occur. The author describes 2 patients who received botulinum toxin type A injections during pregnancy. The first patient was a 34-year-old woman who received 54 units of botulinum toxin type A to treat periorbital lines (crow's feet) unaware that she was 6 weeks pregnant. She had a healthy term baby delivered by cesarean section; Apgar scores were 9/10. The second patient was a 37-year-old woman who received 65 units of botulinum toxin type A to treat facial dynamic lines (forehead, glabelar, and periorbital), unaware that she was 5 weeks pregnant. She had a healthy term baby delivered by cesarean section; Apgar scores were 9/10. The 2 patients were injected only once in the first trimester. Both cases did not have any complications or detectable adverse effects on the fetuses. PMID- 17086004 TI - Transparency in medicine: fact, fiction, or mission impossible? PMID- 17086005 TI - Stress testing and troponin in unstable coronary syndromes: the status trial clinical outcomes and resource use. AB - Cardiac troponins are markers used to diagnose acute myocardial infarction, but their value in guiding management in low- to intermediate-risk patients is not well established. Using a randomized design, the authors compared a strategy using stress testing with blinded troponins vs a troponin I-guided strategy for risk stratification and management of 241 patients with intermediate-risk unstable angina. Fewer stress-tested patients required coronary care unit admission and repeat hospitalization for acute coronary syndrome, at a lower cost. There was no significant difference in rates of death and myocardial infarction due to acute coronary syndrome at 6 months' follow-up. For patients with intermediate-risk acute coronary syndrome, stress testing is as safe as, and more cost-effective than, a troponin I-guided strategy. Patients with marginal troponin I elevations can safely undergo stress testing. Further studies combining stress testing and a troponin I-guided strategy are warranted. PMID- 17086006 TI - Physiologic basis for the surgical treatment of ischemic mitral regurgitation. AB - Ischemic mitral regurgitation (MR) can complicate severe coronary artery disease and myocardial infarction. Ischemic MR results from left ventricular remodeling after myocardial infarction and can also accompany acute myocardial ischemia. The most common mechanism of ischemic MR is Carpentier's type IIIb dysfunction due to an apical and lateral displacement of papillary muscles leading to a tethering of the mitral leaflets. This apical tenting of the leaflets prevents the free margin from reaching the plane of the annulus, significantly reduces the surface of coaptation, and causes MR. Recent advances in imaging studies have led to a better understanding of the pathophysiology of this condition as well as to the development of innovative surgical approaches to treat this disease. Current research efforts have mainly focused on 2 directions: (1) percutaneous approaches to correct MR, and (2) surgical therapy to address the ventricular component of the disease. In this article, the authors define ischemic MR and review its pathophysiology, current management strategies, and future directions. PMID- 17086007 TI - Medical devices and pharmaceuticals: differences in public acceptance. AB - In the past 15 years, rapid technologic advances have introduced medical devices for clinical use that have transformed the care of patients with chronic diseases, especially heart disease. These devices have become truly lifesaving, and their effectiveness has been demonstrated in randomized clinical trials and cost-effectiveness studies. At the same time, a remarkable number of pharmaceutical agents with new actions and capabilities have also been introduced into clinical use. These new compounds have altered the practice of oncology and the care of patients with hematologic malignancies, sometimes with significant improvement in survival. There have been other drugs introduced to manage chronic diseases that alter quality of life but do not usually threaten survival. Recent problems related to the COX-2 anti-inflammatory drugs and ongoing controversies associated with them have raised public awareness; however, there is even more concern about device failures and recalls that have occurred during the same time frame. There is limited information as to why there is such an apparent difference in public opinion toward adverse drug events compared with device problems that lead to advisories or recalls. Among possible reasons are that an implantable device that requires a catheter-based surgical procedure, with its need for hospitalization and detailed explanation about all the potential risks, may be emotionally and intellectually processed differently by a patient/family compared with having a change in prescription from one drug to another. PMID- 17086008 TI - Osler, cardiac disease, and students of medicine--Columbus, OH: December 1899. AB - "Our Distinguished Visitor" was the headline describing the day William Osler honored the Ohio Medical University (OMU) with a visit and clinical lecture. OMU, founded in 1892 and located in downtown Columbus, was the fourth in a series of 5 19th-century Columbus medical schools that were predecessors to The Ohio State University College of Medicine established in 1914. OMU occupied a new 5-story building adjacent to the Protestant Hospital established in 1891. Osler was the guest of Dr Edwin Frazer Wilson, Professor of Therapeutics and Clinical Medicine at OMU, a student of Osler's at the University of Pennsylvania, and an 1885 graduate of that institution. Osler was in Columbus for consultation with one of Dr Wilson's patients. While Osler was in Columbus, a teaching clinic and lecture was arranged at OMU. Murray B. McGonigle, OMU Class of 1900, reported the visit in The Phagocyte, the OMU student publication, and in the Columbus Medical Journal. There was an assemblage of hundreds of students in the amphitheater at clinic hour when OMU Chancellor, Dr David N. Kinsman, introduced Osler. The patient, a 16-year-old young man, was brought to the amphitheater with a history of kidney trouble. Osler's clinical evaluation, physical examination, diagnosis, and discussion of prognosis and treatment provided an object lesson that was impressed on the minds of the students. Following the clinic, Osler addressed the students in his familiar style, demonstrating his easy rapport with students. His comments to the students are classic statements combining personal encouragement with professional attitudes. McGonigle concluded, "these remarks by so eminent an authority are certainly encouraging for the future of the medical student if he will but heed the excellent advice." PMID- 17086009 TI - Cost-effectiveness of impedance cardiography testing in uncontrolled hypertension. AB - To evaluate the short- and long-term cost-effectiveness of impedance cardiography (ICG) testing in uncontrolled hypertensives, we analyzed the Consideration of Noninvasive Hemodynamic Monitoring to Target Reduction of Blood Pressure Levels (CONTROL) trial results that compared the blood pressure-lowering effects of standard vs ICG care. Short-term cost-effectiveness was evaluated as the incremental cost per incremental mm Hg reduced during the trial. Long-term cost effectiveness was evaluated as incremental cost per quality-adjusted life-year gained over 10 years. ICG care short-term cost-effectiveness was 20 US dollar per incremental mm Hg reduced for systolic blood pressure (vs standard care, 36 US dollar per mm Hg reduced) and 23 US dollar per incremental mm Hg reduced for diastolic blood pressure (vs standard care, 79 US dollar per mm Hg reduced). In the long term, ICG resulted in a 476 US dollar cost savings and 0.109 quality adjusted life-years gained per patient (-4,371 US dollar per quality-adjusted life-year gained, sensitivity analysis -8,764 to 13,163 US dollar). The use of ICG testing to reduce blood pressure in uncontrolled hypertensive patients is cost-effective from both a short- and long-term perspective. PMID- 17086010 TI - Multislice CT angiographic reconstruction of marginal artery of Drummond as collateral to occluded superior mesenteric artery. PMID- 17086011 TI - Center of rotation errors: too important to miss. PMID- 17086012 TI - A patient with chest pain and ST elevation: you can't judge a book by its cover! PMID- 17086013 TI - Atrioventricular junctional tachycardia with rate (acceleration)-dependent aberrancy. PMID- 17086014 TI - Pathophysiology and management of syncope in Kearns-Sayre syndrome. AB - A 20-year-old woman with known Kearns-Sayre syndrome was transferred to the emergency department due to syncopal episodes. The electrocardiogram on admission showed complete atrioventricular block. The diagnosis of mitochondrial encephalomyopathy was made when she was 14 years old. At the time of the initial diagnosis, she displayed a normal electrocardiogram pattern. At the age of 17, electrocardiogram recordings demonstrated right bundle branch block with left anterior fascicular block and a prolonged QTc interval of 485 milliseconds (Figure). She was taking coenzyme Q10, oral nicotinamide adenine dinucleotide (reduced), piribedil, amantadine, and primidone. Transthoracic echocardiography revealed normal wall motion of both ventricles and mitral valve prolapse without regurgitation. A permanent dual-chamber pacemaker was immediately implanted. PMID- 17086015 TI - Main issues for achieving blood pressure goals. PMID- 17086016 TI - Arterial elasticity and erectile dysfunction in hypertensive men. AB - Erectile dysfunction is a common symptom among hypertensive patients that impairs quality of life and adherence to antihypertensive pharmacologic therapy. It is also associated with cardiovascular risk factors and disease. The Sexual Health Inventory in Men (SHIM) was administered to 105 ambulatory hypertensive patients, and large and small artery elasticity indices were measured. Each variable was examined in a simple linear regression model or 1-way analysis of variance model to determine each variable's relationship with the SHIM score. Variables that were significantly associated with the SHIM score in the univariate models included age, duration of hypertension, peripheral vascular disease, and small artery elasticity. Large artery elasticity was not significantly associated with the SHIM score. In the multivariate model, age, hypertension duration, and peripheral vascular disease were associated with a lower SHIM score. This study demonstrates a relationship between erectile dysfunction and reduced artery elasticity. PMID- 17086017 TI - Effects of whey peptides on cardiovascular disease risk factors. AB - Previous studies have shown that peptides derived from milk proteins can improve blood pressure. Therefore, the authors tested the blood pressure-lowering effects of a hydrolyzed whey protein supplement rich in bioactive peptides. In a 6-week controlled study, 30 prehypertensive or stage 1 hypertensive subjects (blood pressure >or=120/80 mm Hg and 3.3], TG [>170 mg/dL], and LDL [>138 mg/dL] and lowest-quartile HOMA for HDL [<32 mg/dL]). Hypertensive individuals with insulin resistance (high-quartile HOMA) had higher TG and lower HDL and LDL levels compared with insulin-sensitive hypertensives. High-quartile HOMA, TG, and LDL aggregated in hypertensive families, and TG plus HOMA coaggregated. HDL did not show aggregation. In a multivariate logistic regression, the only significant predictor of an individual's HOMA status was a sibling's HOMA status in a model including age, sex, race, and body mass index (odds ratio=9.12; 95% confidence interval, 3.64-23.14; P<.001). Cardiometabolic variables demonstrate heritability in hypertensive families. Further exploration of common genetic susceptibility loci in hypertension involving these factors is warranted. PMID- 17086020 TI - Human soluble leptin receptor concentration in healthy offspring of hypertensive parents. AB - Essential hypertension is associated with increased plasma leptin levels and decreased human soluble leptin receptor (hsLR) concentration. The aim of this study was to determine whether the concentration of hsLR differs among offspring of hypertensive compared with nonhypertensive parents. Subjects in the 2 groups were matched for age, sex, and body mass index. Forty-six (24 male, 22 female; mean age, 18+/-3 years; body mass index, 22.4+/-1.4 kg/m2) healthy offspring of hypertensive parents (group A) and 50 (28 male, 22 female; mean age, 18+/-3.2 years; body mass index, 22.6+/-1.7 kg/m2) healthy offspring of healthy parents (group B) were studied. The hsLR concentration (enzyme-linked immunosorbent assay method) and leptin plasma levels (radioimmunoassay method) were determined in the study population. Plasma leptin levels were significantly higher (10+/-5 vs 6+/-3 ng/mL; P<.001), while hsLR concentration was significantly lower (20+/-7 vs 29+/ 8 U/mL; P<.001) in group A compared with group B. Our findings suggest that offspring of hypertensive parents have significantly higher plasma leptin levels and significantly lower hsLR concentrations compared with healthy offspring of healthy normotensive parents. Further studies are needed to determine the clinical significance of these observations. PMID- 17086021 TI - Endothelial vascular function in hypertensive patients after renin-angiotensin system blockade. AB - It is unclear whether single and combined pharmacologic inhibition of the renin angiotensin-aldosterone system have similar effects on endothelial function and blood pressure (BP). The authors evaluated 63 hypertensive patients divided into 4 groups (hydrochlorothiazide 25 mg/d; irbesartan [IRBE] 150 mg/d; quinapril [QUIN] 20 mg/d; or IRBE 150 mg/d + QUIN 20 mg/d) and 25 healthy normotensive subjects (normal) followed for 12 weeks. Endothelium-dependent dysfunction measured as flow-mediated dilation at Weeks 0 and 12 were: normal, 11.5%+/-2.4% vs 13.5%+/-2.0%; hydrochlorothiazide, 7.3%+/-2.0% vs 12.8%+/-3.1%; QUIN, 7.2%+/ 2.8% vs 13.2%+/-2.1%; IRBE, 7.1%+/-2.8% vs 13.0%+/-2.9%; and IRBE + QUIN, 7.5%+/ 1.9% vs 12.8%+/-3.0%. Nitroglycerin-mediated responses were: normal, 26.0%+/-1.9% vs 24.0%+/-2.5%; hydrochlorothiazide, 17.0%+/-2.2% vs 18.3%+/-2.6%; QUIN, 17.8%+/ 3.2% vs 23.4%+/-3.0%; IRBE, 16.8%+/-3.6% vs 24.7%+/-2.0%; and IRBE + QUIN, 17.3%+/-3.0% vs 25.1%+/-2.5%. Antihypertensive therapy restored BP to normal and improved the endothelium-dependent and -independent dysfunction after renin angiotensin-aldosterone system blockade. In a further finding, the combined effect of angiotensin-converting enzyme inhibition and angiotensin II type 1 receptor blockade was not superior to the action of either of these treatments separately. PMID- 17086022 TI - Prehypertension--what is it and should it be treated? PMID- 17086023 TI - Could hypertension be a consequence of the 24/7 society? The effects of sleep deprivation and shift work. PMID- 17086024 TI - Who should be treated with implantable cardioverter-defibrillators? PMID- 17086025 TI - Preoperative exercise echocardiography and perioperative cardiovascular outcomes in elderly patients undergoing cancer surgery. AB - Limited information is available on the value of exercise echocardiography (EE) for identifying operative risk in elderly patients undergoing noncardiac surgery. The authors investigated the outcome in 221 consecutive patients 75 years and older who had EE before intermediate- or high-risk cancer surgery. Baseline clinical data, postoperative adverse cardiovascular events (PACE), and 30-day mortality were collected from the medical records and the Social Security Death Index. The mean age +/- SD of the group was 78.9+/-3.5 years; 57% were men. Significant cardiovascular abnormalities (by echocardiography) were present in 71.5%. Mean metabolic equivalents +/- SD achieved during exercise was 4.9+/-1.9. EE identified 50 (22.6%) patients with ischemia and/or infarction. Perioperative beta-blockers were used in 80% of patients with coronary artery disease vs 48.5% without coronary artery disease (P<.0001). There were 31 PACE in 26 (11.8%) patients. Atrial fibrillation was the most common PACE, seen in 18 (8.1%) patients, followed by congestive heart failure in 8 (3.6%), acute coronary syndrome in 2 (0.9%), and cardiac arrest in 1 (0.5%). Thirty-day mortality was only 0.9%. Hospital lengths of stay for patients with and without PACE were 16.8+/-14.9 and 8.5+/-4.8 days (P< or =.0001), respectively. An abnormal EE predicted PACE (22% vs 8.8%; P< .025). Perioperative beta-blockers reduced the incidence of postoperative atrial fibrillation (4.9% vs 12.2%; P=.052). In conclusion, EE is feasible in very elderly patients. There is a high prevalence of cardiac abnormalities in the elderly. An abnormal EE predicts PACE, which, in turn, is associated with increased length of stay. PMID- 17086026 TI - Effects of long-term and ongoing cardiac rehabilitation in elderly patients with coronary heart disease. AB - The authors assessed the effect of very long-term cardiac rehabilitation (over 1 year) on exercise tolerance and coronary heart disease risk factors in patients older than 80 years with coronary heart disease relative to an elderly cohort without coronary heart disease. Included in this retrospective analysis were patients older than 80 at the time of their last evaluation who had completed at least 1 year of a cardiac prevention and exercise training program. A total of 43 coronary heart disease patients and 15 age-matched healthy subjects were included in this study. Long-term cardiac rehabilitation in very elderly coronary heart disease patients was shown to be feasible and effective in improving functional capacity (+5.4% after 1 year; P< .05) and risk factor control, and slowing the decline in exercise tolerance that occurs with normal aging. PMID- 17086027 TI - Enrollment and maintenance of elderly patients in cardiovascular clinical trials. AB - This review addresses the paucity of data available to guide clinical cardiovascular management at advanced elderly age, a likely contributor to the lesser use of beneficial therapies in this population. Cardiovascular disease is the largest health burden of aged persons; this mandates inclusion of elderly persons in clinical research studies of preventive interventions, diagnostic procedures, pharmacotherapy, and high-technology interventional procedures to ascertain their applicability to patients of advanced age. Relevant clinical outcomes include improvement in functional independence and quality of life, decrease in costly hospitalizations, and prolongation of meaningful life. PMID- 17086028 TI - Implantable cardioverter-defibrillators: expectations in the Brazilian elderly. AB - By 2025, Brazil will have the sixth largest elderly population in the world. Thirty percent of the elderly currently have private health insurance and 70% have their health care paid for by the Unified Health System. The system, founded through a governmental entity, provides free health service to all citizens. In 2003, the government passed the Elderly Statement Law, claiming that the elderly have equal access to prevention, promotion, protection, and recovery of health. In spite of the high costs, the placement of implantable cardioverter defibrillators (ICDs) in the elderly is growing. There is a significant regional difference in the number of ICD placements, and there is also a significant sex bias: 73% of the ICDs were placed in elderly men. The majority of ICDs were placed in the elderly from ages 65 to 79 years. Nevertheless, 9.5% of ICDs were placed in octogenarians and nonagenarians, reflecting the valuation of biologic age rather than chronologic age. PMID- 17086029 TI - Implantable cardioverter-defibrillators should be used routinely in the elderly. AB - Implantable cardioverter-defibrillators (ICDs) reduce mortality in selected high risk populations by reducing the risk of sudden death. No clinical trial examining ICD efficacy has focused specifically on benefits and risks in the elderly, but all trials have included substantial numbers of older patients. The author addresses several issues: Is the risk of sudden death different for the elderly than in younger patients? Is ICD implantation feasible in the elderly? Is there evidence that ICD use in the elderly improves survival? The author's research indicates that while total mortality is higher for the elderly, there does not appear to be a specific predilection for arrhythmic death. Recent reports indicate that while ICDs can be implanted in a Medicare population, ICD implantation is associated with significant mortality (0.9%), as well as periprocedural complications (10.8%). In spite of the risks, both secondary and primary prevention trials demonstrate that the survival benefit associated with ICD use in the study populations was at least as great for elderly patients as for younger persons. Thus, published data support judicious ICD use in elderly patients at high risk for sudden death. PMID- 17086030 TI - Why cardioverter-defibrillator implantation might not be the best idea for your elderly patient. AB - Over the last decade, implantable cardioverter-defibrillators (ICDs) have become a part of mainstream clinical practice, with improvements in survival in a broad range of patients. Given the overall limited life expectancy of elderly patients, their frequent comorbidities, and the complications associated with ICD implantation, it may not be reasonable to expect results with ICD implantation in the elderly to be comparable to that found in the clinical trials. In this paper, the authors review the available scientific literature and discuss why mainstream ICD use in the elderly may not be advisable. PMID- 17086031 TI - Chagas' disease and the use of implantable cardioverter-defibrillators in Brazil. AB - The authors discuss the epidemiology and pathogenesis of Chagas' disease in Brazil, including the use of treatment with a cardioverter-defibrillator in patients with low ejection fraction. Select patients may benefit from resynchronization therapy associated with cardioverter-defibrillator treatment. Electrophysiologic study is indicated in the assessment of the potential utility of an implantable cardioverter-defibrillator. PMID- 17086032 TI - The great escape: junctional escape-capture bigeminy. AB - Escape-capture bigeminy is a bigeminal rhythm in which each escape beat is followed by a captured beat. This dysrhythmia is very rare, because its manifestation requires the sinus interval to be longer than the escape interval. This is possible only with severe sinus nodal disease, where the intrinsic sinus rate is extremely low, or with a sinus rhythm associated with an accelerated junctional rhythm. The authors review the case of a 75-year-old man who presented with occasional dizziness and near-syncopal episodes. He was diagnosed with escape-capture bigeminy and subsequently underwent pacemaker placement. PMID- 17086033 TI - Ethical issues in the management of geriatric cardiac patients. PMID- 17086034 TI - Cardiovascular disease preventive measures for the older patient: an epidemiologic perspective. PMID- 17086035 TI - Isolated aortic valve replacement without coronary bypass for aortic valve stenosis involving a congenitally bicuspid aortic valve in a nonagenarian. PMID- 17086036 TI - Are we ready for a statin on our toast? PMID- 17086037 TI - "Giant" coronary aneurysm. PMID- 17086038 TI - Posterior fascicular ventricular tachycardia as an escape rhythm. PMID- 17086039 TI - Impact of International Medical Graduates on U.S. and Global Health Care. Proceedings of the 50th Anniversary Invitational Conference of the Educational Commission for Foreign Medical Graduates. July 21-22, 2006. Philadelphia, Pennsylvania, USA. PMID- 17086040 TI - The role and contributions of IMGs: a U.S. perspective. AB - At 25% of the nation's physician workforce, international medical graduates (IMGs) contribute significantly to the U.S. health care system. Beyond their sheer numbers, however, IMGs have played critically important roles, both in aggregate and as individuals. By choosing to pursue specialties less attractive to U.S. medical graduates, IMGs have filled important gaps that otherwise would have seriously compromised the effectiveness of the U.S. health care system. Moreover, individual IMGs have made notable contributions to the improvement of clinical practice, to biomedical and health services research, and to medical education. The Educational Commission for Foreign Medical Graduates (ECFMG), through its certification process, has enabled the best and the brightest medical students from other nations to train in the United States and can take justifiable pride in the undeniably positive impact IMGs have had on U.S. health care. It is imperative to note, however, that while the United States and other developing nations have benefited enormously from this "medical migration," there is considerable concern about the damaging effects on many countries in the developing world. Among the options to consider in offsetting the negative consequences of this so-called brain drain is working to improve the medical education available to aspiring physicians in "donor" countries. In facilitating such a process, the ECFMG and its partner, the Foundation for Advancement of International Medical Education and Research, have the opportunity to amplify their contributions significantly by building on their established programs of international assistance in improving medical education. PMID- 17086041 TI - International medical education and future directions: a global perspective. AB - Internationalization, one of the most important forces in higher education today, presents a powerful challenge and an opportunity for medical schools. Factors encouraging internationalization include (1) globalization of health care delivery, (2) governmental pressures, (3) improved communication channels, (4) development of a common vocabulary, (5) outcome-based education and standards, (6) staff development initiatives, and (7) competitiveness and commercialization. A three-dimensional model--based on the student (local or international), the teacher (local or international), and the curriculum (local, imported, or international)-offers a range of perspectives for international medical education. In the traditional approach to teaching and learning medicine, local students and local teachers use a local curriculum. In the international medical graduate or overseas student model, students from one country pursue in another country a curriculum taught and developed by teachers in the latter. In the branch-campus model, students, usually local, have an imported curriculum taught jointly by international and local teachers. The future of medical education, facilitated by the new learning technologies and pedagogies, lies in a move from such international interconnected approaches, which emphasize the mobility of students, teachers, and curriculum across the boundaries of two countries, to a transnational approach in which internationalization is integrated and embedded within a curriculum and involves collaboration between a number of schools in different countries. In this approach, the study of medicine is exemplified in the global context rather than the context of a single country. The International Virtual Medical School serves as an example in this regard. PMID- 17086042 TI - Impact of International Medical Graduates on U.S. and Global Health Care: summary of the ECFMG 50th anniversary invitational conference. PMID- 17086043 TI - From defending the walls to improving global medical education: fifty years of collaboration between the ECFMG and the NBME. AB - The author outlines the intertwining roles of the Educational Commission for Foreign Medical Graduates (ECFMG), which is celebrating its 50th anniversary in 2006, and the National Board of Medical Examiners (NBME) in meeting needs for assessment of international medical graduates. Both organizations had early histories focused on a protective role: ensuring that only the most qualified foreign-trained doctors could train or practice in the United States. The two organizations have interacted throughout the ECFMG's 50-year history to improve the assessment of internationally trained doctors. As both the ECFMG and the NBME have matured, their missions have expanded to include improvement of medical education and assessment around the world. Much of the success of each organization in fulfilling its mission can be attributed to their close collaboration through the past 50 years. PMID- 17086044 TI - The future of medical licensure in the United States. AB - Medical licensure in the United States is undergoing significant change. With calls for greater accountability and transparency, state medical boards and their membership association, the Federation of State Medical Boards (FSMB), are seeking ways to assure the public that physicians are maintaining their competence throughout the lifetime of their practice of medicine. At present, competence in cognitive, clinical, and communicative skills is regularly measured only at initial licensure. Yet, the public and policy-related organizations are demanding ongoing assessment of physicians' ability to safely and competently practice medicine. The author reports on activities that involve the FSMB and other national organizations, including the Educational Commission for Foreign Medical Graduates, in planning for a future of increased accountability and transparency of the licensing and regulatory communities that oversee the practice of medicine. He notes that topics of discussion include possible nationalization of what has been traditionally state-based licensure. He raises questions about a future that may include specialty-based licensure and greater national and even international license portability. PMID- 17086045 TI - International medical graduates in the United States: a view from an ECFMG certificant. AB - The author was instilled with a passion for mathematics and physics by his father, who taught those subjects in a small Algerian town. Another indelible influence came during a high school mathematics class when his teacher gave the class a problem to solve. Little did the students know that it was Fermat's Last Theorem, which stumped them, and before that, every mathematician since 1630. This experience taught the author that failing to get the final answer was part of learning. He became enchanted with imaging techniques and after earning his medical degree in Algeria, came to study at Johns Hopkins. There he received the training he desired in diagnostic radiology. The author believes science has no borders and would like to see the opportunities that were extended to him in 1975 given to immigrants today. Although the United States produces many graduates in the sciences and mathematics, the nation still has a shortfall and must, he argues, work harder to educate and inspire this country's youth in addition to welcoming the brightest and most able scientists from around the world. He also discusses the crucial role of the National Institutes of Health in furthering global health by funding international biomedical research and by transforming medicine in the 21st century. PMID- 17086046 TI - Global standards and accreditation in medical education: a view from the WFME. AB - Globalization of medicine is increasing, as manifested by the growing number of migrating doctors and cross-border education providers. In addition, new medical schools of dubious quality are proliferating. This situation accentuates the need to define standards and introduce effective and transparent accreditation systems. With this background, and reflecting the important interface between medical education and health care delivery, a World Health Organization (WHO)/World Federation for Medical Education (WFME) Strategic Partnership to improve medical education was formed in 2004. In addition to working on reform processes, capacity building, and evaluation of medical education at the regional and national levels, the partnership in 2005 published guidelines for accreditation of basic medical education. Only a minority of countries have quality assurance systems based on external evaluation, and most of these use only general criteria for higher education. The WHO/WFME Guidelines recommend establishing accreditation that is effective, independent, transparent, and based on criteria specific to medical education. An important prerequisite for this development was the WFME Global Standards program, initiated in 1997 and widely endorsed. The standards are now being used in all regions as a basis for improving medical education throughout its continuum and as a template for national and regional accreditation standards. Promotion of national accreditation systems will pivotally influence future international appraisal of medical education. Information about accreditation status--agencies involved and criteria and procedures used--will be essential to future databases of medical schools and will be a foundation for international "meta-recognition" of institutions and programs ("accrediting the accreditors"). PMID- 17086047 TI - The circle game: understanding physician migration patterns within Canada. AB - This report explores the movement of physicians to, from, and within Canada and identifies recurring patterns of migration. The primary position of the report is that physician movement is part of reality both internationally and within Canada, and that movement of Canadian-trained physicians creates a need for international medical graduates (IMGs) in "physician-losing" locations. The report's argument is based on data retrieved from public sources on aggregate physician practice patterns in Canada and analyzed for migration patterns. In addition, literature was reviewed on factors affecting the migration patterns being described.Canadian-educated physicians have tended to move from less prosperous to more prosperous provinces and from rural to urban areas; because of the resulting need, the physician-losing locales generally have the highest proportions of IMGs. Physicians traditionally have tended to emigrate from Canada to the United States, thus increasing Canadian demand for IMGs, but recently this movement has slowed and even reversed. In Canada, liberalized immigration policies for physicians combined with a shortage of postgraduate training positions to create a serious bottleneck early in the current decade. However, this problem is now being resolved. In summary, physician migration within Canada shows specific long-term patterns, and IMGs will be needed in underserved areas for years to come. Well-informed policies for workforce management are essential in Canada to ensure an adequate physician supply consisting mainly of Canadian educated physicians but also including IMGs. A role for nonadvocacy groups such as the Educational Commission for Foreign Medical Graduates may be to help ensure that recruitment of physicians from developing countries follows accepted ethical principles. PMID- 17086048 TI - Migration, medical education, and health care: a view from Pakistan. AB - PURPOSE: To review physician flows in Pakistan in the contexts of capacity to educate and provide for anticipated health needs. METHOD: Literature review was used to refine estimates and assumptions about physician supply and demand. Net physician supply was calculated from data on physician inflows and estimates of migration and other causes of physician loss. Projections of anticipated needs up to 2020 were calculated, assuming different levels of physician need or demand. Reasons for emigration were extracted from interviews, questionnaires, and reports; health indicators and migration figures were obtained from literature and Web sites. RESULTS: Approximately 74,000 physicians were practicing in Pakistan in 2005. Annually, local medical schools and international medical graduate certification provide 5,400 physicians, soon to reach 6,800; 1,150 physicians emigrate; and an estimated 570 physicians stop practicing for various reasons. The current ratio (0.473) of physicians to 1,000 population is inadequate to maintain the nation's health. Future Physician Workforce Shortages (PWSs) for Pakistan range between 57,900 and 451,102 physicians in 2020, depending on assumptions about future need. CONCLUSIONS: Pakistan cannot meet its needs for health care given the current levels of production and dependency on physicians in the organization of the system. Although outmigration contributes to the problem, it is the growing demand for health care from increases in population; adverse conditions that generate ill-health; and increasing expectations that dramatically increase the PWSs. The anticipated shortfall in Pakistan reflects a global pattern. Definition of physician roles and improvement of standards through refined assessment require serious consideration. PMID- 17086049 TI - Concurrent sessions: exploring issues relating to international medical graduates. PMID- 17086051 TI - Celebrating 50 years of experience: an ECFMG perspective. AB - As indicated in its mission statement, the Educational Commission for Foreign Medical Graduates (ECFMG), established in 1956, "promotes quality health care for the public by certifying IMGs for entry into U.S. graduate medical education, and by participating in the evaluation and certification of other physicians and health care professionals. In conjunction with its Foundation for Advancement of International Medical Education and Research, and other partners, it actively seeks opportunities to promote international medical education through programmatic and research activities." From 1958 through 2005, the ECFMG certified 287,382 international medical graduates (IMGs); the number per year has varied, depending on the world political situation and other factors. Currently, IMGs constitute 25% of practicing physicians in the United States. India, followed by the United States and the Philippines, has provided the largest number of certified IMGs. The ECFMG has developed extensive resources for verifying medical school diplomas and other credentials of applicants, and it provides credentials verification services to other organizations. Under the U.S. government J-1 Exchange Visitor Program, it also has played a major role in facilitating the entry of foreign physicians to the United States for clinical training and research experience. Over the years, testing of IMGs seeking ECFMG Certification has evolved; whereas initially applicants took examinations specifically for IMGs, they now take the same United States Medical Licensing Examination (USMLE) sequence of examinations as U.S. medical graduates. Passage of the USMLE is required for ECFMG Certification and licensure in all jurisdictions. PMID- 17086052 TI - AIDS vaccine development and challenge viruses: getting real. PMID- 17086053 TI - Acceptability of Carraguard vaginal gel use among Thai couples. AB - OBJECTIVES: To evaluate the acceptability of candidate microbicide Carraguard among couples participating in a safety trial. STUDY DESIGN: A 6-month randomized, placebo-controlled trial was conducted in sexually active, low-risk couples in Thailand. METHODS: Couples who were monogamous, HIV uninfected, and not regular condom users were enrolled. Acceptability data were collected through structured questionnaires at repeated intervals. At the closing study visit, participants were asked questions about hypothetical product characteristics and future use. Compliance with gel use was assessed by questionnaires, coital diaries, and tracking of used and unused applicators. RESULTS: Among 55 enrolled couples, follow up and adherence with gel use were high and sustained, with 80% of women using gel in over 95% of vaginal sex acts. Because acceptability results from Carraguard and placebo arms were similar, they were combined for this analysis. Overall, 92% of women and 83% of men liked the gel somewhat or very much; 66% of women and 72% of men reported increased sexual pleasure with gel use; and 55% of women and 62% of men reported increased frequency of intercourse. Only 15% of women but 43% of men thought that gel could be used without the man knowing. Although men and women had similar views overall, concordance within couples was low, with no kappa coefficients above 0.31. CONCLUSION: Carraguard gel use was acceptable to low-risk couples in northern Thailand. Reported associations between gel use and increased sexual pleasure and frequency suggest a potential to market microbicide products for both disease prevention and enhancement of pleasure. PMID- 17086054 TI - Replicating Ad-recombinants encoding non-myristoylated rather than wild-type HIV Nef elicit enhanced cellular immunity. AB - OBJECTIVE: To determine if immunization with non-myristoylated nef would elicit enhanced cellular immune responses resulting from improved presentation of Nef peptides by MHC-I on the cell surface, and enhanced T-cell help. DESIGN: The myristoylation site of HIV and SIV Nef is required for several Nef functions that modulate the immune response in an infected host, including downregulation of MHC I, MHC-II, and CD4, and increased expression of the invariant chain on the cell surface. We constructed replication-competent Ad5- and Ad7-HIV recombinants encoding wild-type nef (nefWT) or a nef mutant (nefNM) lacking 19 amino-terminal amino acids, including the myristoylation site, and sequentially immunized chimpanzees mucosally, first with Ad5-HIVnef recombinants and subsequently with Ad7-HIVnef recombinants. METHODS: Peripheral blood lymphocytes were evaluated over the immunization course for Nef-specific cellular immune responses by interferon (IFN)-gamma ELISPOT and T-cell proliferation assays. Nef-specific CD4 and CD8 memory T cells that produced intracellular IFN-gamma, interleukin-2, and tumor necrosis factor (TNF)-alpha were assessed by flow cytometry. RESULTS: In comparison to immunization with Ad-HIVnefWT, Ad-HIVnefNM elicited statistically significant increases in numbers of IFN-gamma-secreting cells after the Ad7 HIVnefNM immunization and increased T-cell proliferative responses following both Ad5- and Ad7-HIVnefNM immunizations. Nef-specific CD4 and CD8 memory T-cell populations secreting TNF-alpha were also significantly increased in the Ad HIVnefNM immunization group. CONCLUSIONS: The results support the hypothesis that immunization with Ad-recombinants encoding HIVnefNM rather than HIVnefWT elicits enhanced cellular immunity resulting from improved antigen presentation and greater T-cell help. PMID- 17086055 TI - Ezetimibe, a promising lipid-lowering agent for the treatment of dyslipidaemia in HIV-infected patients with poor response to statins. AB - OBJECTIVE: To assess the efficacy, safety, and pharmacokinetic interactions of ezetimibe in HIV-infected patients with poorly controlled antiretroviral associated dyslipidaemia while taking pravastatin alone. DESIGN: A prospective, open-label, one-arm study of 24 weeks duration. PATIENTS AND SETTING: Nineteen patients (18 on stable HAART), with low density lipoprotein (LDL)-cholesterol values of > or = 130 mg/dl despite the use of pravastatin. METHODS: Ezetimibe, 10 mg/day, was added to pravastatin 20 mg/day, while patients maintained the same antiretroviral regimen. Determinations of total, LDL-, and high density lipoprotein (HDL)-cholesterol, triglycerides, apoproteins, and inflammatory factors (homocystein and C-reactive protein) were performed at baseline, and at weeks 6, 12, and 24. Liver enzymes and creatinine phosphokinase were also assessed. Protease inhibitor (PI) or non-nucleoside reverse transcriptase inhibitor (NNRTI) Cmin was determined just before and 12 weeks after ezetimibe introduction. RESULTS: At week 24, 61.5% of patients achieved the endpoint of the study (LDL-cholesterol < 130 mg/dl). Significant declines in mean total and LDL cholesterol levels were observed between baseline and weeks 6, 12, and 24, irrespective of antiretroviral type (PI or NNRTI). Mean HDL-cholesterol and apoprotein A increased significantly. No patients discontinued therapy due to intolerance or presented toxicity of grade 2 or more. No differences were observed in lopinavir or nevirapine Cmin measured just before and 12 weeks after ezetimibe introduction. CONCLUSION: The addition of ezetimibe to ongoing pravastatin seems to be an effective and safe option for HIV-infected patients not achieving the NCEP ATPIII LDL-cholesterol goals while receiving a statin alone. Its high tolerability and the lack of interactions with the cytochrome CYP3A4 indicate that ezetimibe will not increase the risk of toxicity or pharmacokinetic interactions with antiretrovirals. PMID- 17086056 TI - Antiretroviral therapy and the prevalence of osteopenia and osteoporosis: a meta analytic review. AB - INTRODUCTION: Prevalence estimates of osteopenia and osteoporosis (reduced bone mineral density; BMD) in HIV-infected patients and the role of antiretroviral therapy (ART) varies in the literature. METHODS: We conducted a meta-analytical review of cross-sectional studies published in English to determine the pooled odds ratios (OR) of reduced BMD and osteoporosis in the following groups: HIV positive versus HIV-negative; ART-treated versus ART-naive; protease inhibitor (PI)-treated versus PI-untreated. We searched the MEDLINE, PubMed, and EMBASE databases for eligible references between January 1966 and November 2005. Random effects models were used to generate pooled OR estimates and confidence intervals. RESULTS: Of 37 articles identified, 20 met the inclusion criteria. Of the 884 HIV-infected patients, 67% had reduced BMD, of whom 15% had osteoporosis, yielding a pooled OR of 6.4 and 3.7, respectively, compared with HIV-uninfected controls (n = 654) using 11 studies with available data. Compared with ART-naive patients (n = 202, 10 studies), ART-treated individuals (n = 824) had a 2.5-fold increased odds of prevalent reduced BMD. The risk of prevalent osteoporosis (seven studies) was similarly elevated in ART-treated individuals. Compared with non-PI-treated HIV patients (n = 410, 14 studies), PI-treated patients (n = 791) had increased odds of reduced BMD and osteoporosis (12 studies). Few studies adjusted for important covariates such as HIV disease severity or treatment duration. CONCLUSION: The prevalence of osteoporosis in HIV-infected individuals is more than three times greater compared with HIV-uninfected controls. ART exposed and PI-exposed individuals had a higher prevalence of reduced BMD and osteoporosis compared with their respective controls. The influence of other disease and treatment variables on these estimates could not be determined. PMID- 17086057 TI - Histological response to pegIFNalpha-2a (40KD) plus ribavirin in HIV-hepatitis C virus co-infection. AB - OBJECTIVE: Paired liver biopsies from patients enrolled in the multinational AIDS PEGASYS Ribavirin International Co-infection Trial were analysed to investigate a possible correlation between virological and histological responses. DESIGN AND METHODS: A total of 860 HIV-hepatitis C virus (HCV)-co-infected patients were randomly assigned to receive pegIFNalpha-2a (40KD) 180 microg/week plus 800 mg daily ribavirin, pegIFNalpha-2a (40KD) plus placebo or conventional IFNalpha-2a 3 MIU three times a week plus ribavirin for 48 weeks. Paired biopsies were obtained from 401 patients and scored locally using the Ishak-modified histological activity index (HAI). The second biopsy was obtained, on average, 26 weeks or more after the end of treatment. Histological response was defined as a 2-point or greater reduction in the HAI score. RESULTS: The histological response rate was significantly higher in patients receiving pegIFNalpha-2a (40KD) plus ribavirin (57%) than in patients receiving pegIFNalpha-2a (40KD) plus placebo (39%; P < 0.017) or IFNalpha-2a plus ribavirin (41%; P = 0.04). Histological response was correlated with virological response, with the histological response rate ranging from 62 to 74% in patients who achieved a sustained virological response (SVR). Histological response was also seen in 32-43% of patients not achieving an SVR. A higher total HAI score was the only prognostic factor for achieving histological response. CONCLUSION: The histological response rate was significantly higher in HIV-HCV-co-infected patients who received pegIFNalpha-2a (40KD) plus ribavirin than in those receiving pegIFNalpha-2a (40KD) plus placebo or IFNalpha-2a plus ribavirin. Histological response was correlated with virological response, although a substantial proportion of patients who did not achieve an SVR experienced histological improvement. PMID- 17086058 TI - Long-term outcome of AIDS-associated cryptococcosis in the era of combination antiretroviral therapy. AB - BACKGROUND: Immune restoration following combination antiretroviral therapy (cART) questions the maintenance of prophylaxis among HIV-infected patients with cryptococcosis. OBJECTIVE: To describe the long-term outcome after the diagnosis of cryptococcosis at the cART era. DESIGN: Multicentre cohort of patients with a diagnosis of cryptococcosis between 1996 and 2000, follow-up until December 2002. Comparison with a historical cohort (1990-1994) for survival. SETTING: Eighty four French AIDS clinical centres. PATIENTS: Two-hundred and forty HIV-infected adult patients at the cART era and 149 at the pre-cART era experiencing a first episode of culture-confirmed cryptococcosis. RESULTS: In the cART era, 82/189 patients surviving more than 3 months after initiation of antifungal therapy had their maintenance therapy interrupted with a subsequent median follow-up of 19 months. Their relapse rate per 100 person-years was 0.9 [95% confidence interval (CI),0.0-2.0]. When considering the whole cART cohort, probability of reaching negative serum cryptococcal antigen was 71% after 48 months of follow-up. A CD4 cell count < 100/microl [relative risk (RR), 5.5; 95% CI, 1.3-22.2], antifungal therapy < 3 months over the past 6 months [RR, 5.0; 95% CI, 1.1-22.3] and serum cryptococcal antigen titre > or = 1/512 [RR, 3.5; 95% CI, 1.1-10.8] were associated with a higher rate of cryptococcosis relapse. The mortality rate per 100 person-years was 15.3 [95% CI,12.2-18.4] in the cART era versus 63.8 [95% CI,53.0-74.9] in the pre-cART era although early mortality did not differ between the two periods. CONCLUSION: Overall survival after cryptococcosis has dramatically improved at the cART era. Immune restoration and low serum cryptococcal antigen titres are associated with lower cryptococcosis relapse rates. PMID- 17086059 TI - Routine surveillance for the detection of acute and recent HIV infections and transmission of antiretroviral resistance. AB - OBJECTIVE: To estimate the rate of acute and recent HIV infections and the prevalence of primary antiretroviral resistance. DESIGN, SETTING, AND SUBJECTS: A consecutive sample of individuals presenting for HIV testing at the San Francisco municipal sexually transmitted diseased (STD) clinic in 2004 (n = 3789). MAIN OUTCOME MEASURES: HIV antibody-positive specimens were screened by BED IgG capture enzyme immunoassay to identify recent infections. HIV antibody-negative specimens were screened by nucleic acid amplification testing (NAAT) to detect acute infections. Newly detected infections were genotyped to detect primary antiretroviral resistance. RESULTS: There were 11 acute and 44 recent HIV infections among the total 136 newly detected cases. NAAT increased case identification by 8.08% over standard antibody testing. Acute HIV infections were associated with having a known HIV-positive partner, and a history of hepatitis B, syphilis, and chlamydia. The prevalence of primary antiretroviral resistance was 13.2%, with drug-resistant mutations detected in 17 of 129 cases genotyped. Mutations conferring resistance to non-nucleoside reverse transcriptase inhibitors (NNRTI) were present in 11 of 17 cases. CONCLUSION: The integration of HIV nucleic acid amplification, recent infection, and antiretroviral resistance testing enhanced HIV/STD surveillance. The high proportion of NNRTI mutations detected suggests they may be more common in source partners or more fit for transmission than other forms of drug-resistant HIV-1. Primary antiretroviral resistance monitoring in STD clinic patients may guide the selection of treatment and post-exposure prophylaxis regimens active against viruses being transmitted in the community, and provide health departments with surveillance data in a sentinel population at risk of HIV transmission. PMID- 17086060 TI - Population-based surveillance for cryptococcosis in an antiretroviral-naive South African province with a high HIV seroprevalence. AB - OBJECTIVES: To measure the burden of disease and describe the epidemiology of cryptococcosis in Gauteng Province, South Africa. DESIGN AND METHODS: The study was an active, prospective, laboratory-based, population-based surveillance. An incident case of cryptococcosis was defined as the first isolation by culture of any Cryptococcus species from any clinical specimen, a positive India ink cryptococcal latex agglutination test or a positive histopathology specimen from a Gauteng resident. Cases were identified prospectively at all laboratories in Gauteng. Case report forms were completed using medical record review and patient interview where possible. RESULTS: Between 1 March 2002 and 29 February 2004, 2753 incident cases were identified. The overall incidence rate was 15.6/100 000. Among HIV-infected persons, the rate was 95/100 000, and among persons living with AIDS 14/1000. Males and children under 15 years accounted for 49 and 0.9% of cases, respectively. The median age was 34 years (range, 1 month-74 years). Almost all cases (97%) presented with meningitis. Antifungal therapy was given to 2460 (89%) cases of which 72% received fluconazole only. In-hospital mortality was 27% (749 cases). Recurrences occurred in 263 (9.5%) incident cases. Factors associated with death included altered mental status, coma or wasting; factors associated with survival included employment in the mining industry, visual changes or headache on presentation. CONCLUSIONS: This study demonstrates the high disease burden due to cryptococcosis in an antiretroviral-naive South African population and emphasizes the need to improve early recognition, diagnosis and treatment of the condition. PMID- 17086061 TI - Effectiveness and cost-effectiveness of strategies to expand antiretroviral therapy in St. Petersburg, Russia. AB - OBJECTIVE: To assess the effectiveness and cost-effectiveness of treating HIV infected injection drug users (IDUs) and non-IDUs in Russia with highly active antiretroviral therapy HAART. DESIGN AND METHODS: A dynamic HIV epidemic model was developed for a population of IDUs and non-IDUs. The location for the study was St. Petersburg, Russia. The adult population aged 15 to 49 years was subdivided on the basis of injection drug use and HIV status. HIV treatment targeted to IDUs and non-IDUs, and untargeted treatment interventions were considered. Health care costs and quality-adjusted life years (QALYs) experienced in the population were measured, and HIV prevalence, HIV infections averted, and incremental cost-effectiveness ratios of different HAART strategies were calculated. RESULTS: With no incremental HAART programs, HIV prevalence reached 64% among IDUs and 1.7% among non-IDUs after 20 years. If treatment were targeted to IDUs, over 40 000 infections would be prevented (75% among non-IDUs), adding 650 000 QALYs at a cost of USD 1501 per QALY gained. If treatment were targeted to non-IDUs, fewer than 10 000 infections would be prevented, adding 400 000 QALYs at a cost of USD 2572 per QALY gained. Untargeted strategies prevented the most infections, adding 950 000 QALYs at a cost of USD 1827 per QALY gained. Our results were sensitive to HIV transmission parameters. CONCLUSIONS: Expanded use of antiretroviral therapy in St. Petersburg, Russia would generate enormous population-wide health benefits and be economically efficient. Exclusively treating non-IDUs provided the least health benefit, and was the least economically efficient. Our findings highlight the urgency of initiating HAART for both IDUs and non-IDUs in Russia. PMID- 17086062 TI - Predictors of HIV sero-status among drug injectors at three Ukraine sites. AB - OBJECTIVE: To assess the HIV serostatus of injection drug users (IDU) in Ukraine, as well as associations between serostatus and selected demographic and risk factors. DESIGN AND METHODS: IDU were recruited from the streets in Kiev, Odessa and Makeevka/Donesk. Participants were interviewed using an HIV risk behavior assessment and tested for HIV with a finger-stick rapid test. Multiple logistic regression was used to identify determinants of HIV infection. RESULTS: Of the 891 IDUs surveyed, one-third came from each site and 22% were female. Their mean age was 29 years and on average they had been injecting for slightly more than 10 years. Seven hundred and seventy-eight of the total sample did not know their HIV status when first interviewed; they are the participants in this investigation. Overall, 33% tested positive for HIV, including 34% in Kiev, 51% in Odessa and 17% in Makeevka/Donesk. Independent predictors of HIV included injecting a sedative/opiate mixture, female sex, having sex with a person who was HIV positive or whose HIV status was unknown and injecting daily. HIV-negative IDU were significantly younger than those infected, they were more likely to be from Makeevka/Donesk and they were more likely to have been sexually active. CONCLUSIONS: Rates of HIV infection among IDU vary considerably across Ukraine, although even in the site with the lowest rate nearly one in five was infected. The extent of drug and sex-related risk behaviors calls for interventions to reduce the spread of HIV and other infectious diseases. PMID- 17086063 TI - Regression of liver fibrosis in hepatitis C virus/HIV-co-infected patients after treatment with pegylated interferon plus ribavirin. PMID- 17086064 TI - A novel hepatitis B virus mutation with resistance to adefovir but not to tenofovir in an HIV-hepatitis B virus-co-infected patient. AB - A molecular virology analysis performed in an HIV-hepatitis B virus (HBV) co infected patient showed the emergence of an unusual HBV polymerase gene mutation (rt A181T) under adefovir therapy, conferring resistance to adefovir but not to tenofovir, as proved by in-vitro phenotypic analysis. This observation suggests that careful monitoring of co-infected patients is required to diagnose HBV resistance to nucleos(t)ide analogues, and that tenofovir may be active at least against some of the adefovir-resistant strains. PMID- 17086065 TI - Persistence of a sexually transmitted highly resistant HIV-1: pol quasispecies evolution over 33 months in the absence of treatment. AB - The evolution of a sexually transmitted multiresistant HIV-1 in a linked transmission chain was followed for 33 months to assess its potential to persist in the absence of treatment. The multiresistant HIV reverted slowly to wild type in reverse transcriptase (positions 44, 67, 74, 118) rendering the virus only susceptible to lamivudine/emtricitabine. Persistence of the replication-competent resistant HIV increases its potential to spread further and strengthens the importance of resistance testing in newly infected patients. PMID- 17086066 TI - Nevirapine versus efavirenz in 742 patients: no link of liver toxicity with female sex, and a baseline CD4 cell count greater than 250 cells/microl. AB - Recent studies have reported increased nevirapine hepatotoxicity in female patients with CD4 lymphocyte counts greater than 250 cells/microl (especially pregnant women). However, our open-label comparison of 742 patients treated with either nevirapine or efavirenz-based HAART as naive patients, experienced subjects, or patients on salvage therapy, found no increased hepatotoxicity in nevirapine-treated subjects, in particular with regard to both sex (females versus males) and T-cell-mediated immunodeficiency (CD4 cell counts above versus below 250 cells/microl). PMID- 17086067 TI - Immune recovery vitritis in an HIV patient with isolated toxoplasmic retinochoroiditis. PMID- 17086068 TI - Techniques for measuring regional body fat: computed tomography may be suitable for evaluation of facial lipoatrophy. PMID- 17086069 TI - Costing implications of recent treatment interruption studies. PMID- 17086070 TI - HIV and the hidden face of the thymus. PMID- 17086071 TI - Cell phones: tightening the communication gap in resource-limited antiretroviral programmes? PMID- 17086072 TI - Olivocochlear efferents: anatomy, physiology, function, and the measurement of efferent effects in humans. AB - This review covers the basic anatomy and physiology of the olivocochlear reflexes and the use of otoacoustic emissions (OAEs) in humans to monitor the effects of one group, the medial olivocochlear (MOC) efferents. MOC fibers synapse on outer hair cells (OHCs), and activation of these fibers inhibits basilar membrane responses to low-level sounds. This MOC-induced decrease in the gain of the cochlear amplifier is reflected in changes in OAEs. Any OAE can be used to monitor MOC effects on the cochlear amplifier. Each OAE type has its own advantages and disadvantages. The most straightforward technique for monitoring MOC effects is to elicit MOC activity with an elicitor sound contralateral to the OAE test ear. MOC effects can also be monitored using an ipsilateral elicitor of MOC activity, but the ipsilateral elicitor brings additional problems caused by suppression and cochlear slow intrinsic effects. To measure MOC effects accurately, one must ensure that there are no middle-ear-muscle contractions. Although standard clinical middle-ear-muscle tests are not adequate for this, adequate tests can usually be done with OAE-measuring instruments. An additional complication is that most probe sounds also elicit MOC activity, although this does not prevent the probe from showing MOC effects elicited by contralateral sound. A variety of data indicate that MOC efferents help to reduce acoustic trauma and lessen the masking of transients by background noise; for instance, they aid in speech comprehension in noise. However, much remains to be learned about the role of efferents in auditory function. Monitoring MOC effects in humans using OAEs should continue to provide valuable insights into the role of MOC efferents and may also provide clinical benefits. PMID- 17086073 TI - Effects of programming threshold and maplaw settings on acoustic thresholds and speech discrimination with the MED-EL COMBI 40+ cochlear implant. AB - OBJECTIVES: The principal task in the programming of a cochlear implant (CI) speech processor is the setting of the electrical dynamic range (output) for each electrode, to ensure that a comfortable loudness percept is obtained for a range of input levels. This typically involves separate psychophysical measurement of electrical threshold ([theta] e) and upper tolerance levels using short current bursts generated by the fitting software. Anecdotal clinical experience and some experimental studies suggest that the measurement of [theta]e is relatively unimportant and that the setting of upper tolerance limits is more critical for processor programming. The present study aims to test this hypothesis and examines in detail how acoustic thresholds and speech recognition are affected by setting of the lower limit of the output ("Programming threshold" or "PT") to understand better the influence of this parameter and how it interacts with certain other programming parameters. DESIGN: Test programs (maps) were generated with PT set to artificially high and low values and tested on users of the MED-EL COMBI 40+ CI system. Acoustic thresholds and speech recognition scores (sentence tests) were measured for each of the test maps. Acoustic thresholds were also measured using maps with a range of output compression functions ("maplaws"). In addition, subjective reports were recorded regarding the presence of "background threshold stimulation" which is occasionally reported by CI users if PT is set to relatively high values when using the CIS strategy. RESULTS: Manipulation of PT was found to have very little effect. Setting PT to minimum produced a mean 5 dB (S.D. = 6.25) increase in acoustic thresholds, relative to thresholds with PT set normally, and had no statistically significant effect on speech recognition scores on a sentence test. On the other hand, maplaw setting was found to have a significant effect on acoustic thresholds (raised as maplaw is made more linear), which provides some theoretical explanation as to why PT has little effect when using the default maplaw of c = 500. Subjective reports of background threshold stimulation showed that most users could perceive a relatively loud auditory percept, in the absence of microphone input, when PT was set to double the behaviorally measured electrical thresholds ([theta]e), but that this produced little intrusion when microphone input was present. CONCLUSIONS: The results of these investigations have direct clinical relevance, showing that setting of PT is indeed relatively unimportant in terms of speech discrimination, but that it is worth ensuring that PT is not set excessively high, as this can produce distracting background stimulation. Indeed, it may even be set to minimum values without deleterious effect. PMID- 17086074 TI - Outcomes of long-term outpatient tinnitus-coping therapy: psychometric changes and value of tinnitus-control instruments. AB - OBJECTIVES: Increasing tinnitus compliance and coping are desirable aims of successful treatment in chronic tinnitus. However, application of established procedures such as tinnitus retraining therapy (TRT) is often relatively short. In addition, the value of tinnitus control instruments (TCI) is questionable, especially for minor severity levels of tinnitus, and the comparability of treatment results is low. To evaluate long-term changes of tinnitus-related distress, defined psychometric data were collected in patients with compensated tinnitus (cT) or decompensated tinnitus (dT) during a standardized 2-yr outpatient tinnitus-coping therapy (TCT). DESIGN: In a prospective clinical investigation, the data of 70 tinnitus patients were recorded at the beginning and at 6-mo intervals, with a final investigation after 24 mo. The first group consisted of 40 patients with cT and dT who were randomly assigned to a treatment group and a waiting-list control group. After a period of 12 mo without treatment, the control group was treated similarly. The tinnitus questionnaire (TQ) of Goebel and Hiller, visual analog scales (VAS), and a severity questionnaire for tinnitus-associated complaints were used as psychodiagnostic instruments. Therapy components consisted of counseling, fitting patients with TCIs (TCI provision), auditory and relaxation training, and psychosomatic care if necessary. A second group of 30 patients with cT receiving TCT without TCI devices was investigated to evaluate the additive efficacy of TCI in cT. RESULTS: The initial TQ score did not differ between the treatment group and the waiting list control group. After 12 mo, the control group did not show any significant changes, but the treatment group had improved significantly. During TCT, the combined data of both groups showed a statistically significant decrease of the TQ score in dT (59.1 to 34.8) and cT (32.8 to 24.0). These changes were especially reflected by the subscales of cognitive and emotional distress and also by the VAS and the severity questionnaire. dT patients benefitted without dependence on age or duration of pre-existing tinnitus; for cT patients, this was true mainly of the younger and older subjects and for tinnitus lasting for less than 1 yr. TCI provision improved the efficacy of TCT in patients with cT. CONCLUSIONS: The psychometric changes demonstrate a clear decrease of tinnitus related distress in all severity levels of sufficiently treated chronic tinnitus. Long-term TCT induces improvement even up to the time of 24 mo. With TCIs established particularly in patients with dT, our results suggest that a supporting adjustment of TCI devices is helpful in cT to foster quicker rehabilitation. The outpatient interdisciplinary TCT, consisting of cognitive tinnitus desensitization, TCI provision, and psychosomatic support if required, represents a successful treatment strategy for both dT and cT patients. PMID- 17086076 TI - Psychophysical assessment of spatial spread of excitation in electrical hearing with single and dual electrode contact maskers. AB - OBJECTIVE: To evaluate psychophysically the spatial spread of excitation in electrical hearing with a new dual contact masker and to investigate under which conditions it is possible to stimulate fibers in the immediate neighborhood of an electrode contact, which were not excited by neighboring electrode contacts. DESIGN: In this study a psychophysical forward masking paradigm with a dual contact masker was used to avoid off-site listening, the electrical analogue of off-frequency listening. The masker stimulus (300 msec) is presented nonsimultaneously on two electrode contacts, one on the apical side and another on the basal side of the probe contact, followed by a probe stimulus of 20 msec.Unmasked probe thresholds were compared with masked ones at a number of masker-probe distances, whereas growth of masking curves were measured for a fixed masker contact pair. Standard selectivity measurements (single contact masking) and the recovery of forward masking with one masker contact were included for comparison with existing methods. All experiments were carried out with six participants who use the Clarion CII device with a HiFocus I electrode array. RESULTS: For dual contact masking the amount of masking was significantly greater than for single contact masking and the width of the masking patterns was on average 1.1 mm broader than for single contact masking, resulting in a broad region of excitation, with masker-probe overlap for distances greater than 3 mm. Masking widths for dual and single contact masking were highly correlated. Growth of masking curves were highly nonlinear. They showed a strong elevation of the slope that starts for most subjects around the middle of the dynamic range or above. For 4 out of 6 subjects, no probe threshold was found above a masker amplitude of about 400-500 microA. The ratio of the maximum measurable masked probe thresholds and unmasked probe threshold ranged from 1.7 to 2.6 (S4 excluded). Recovery of masking functions follow an exponential decay. Time constants tau for the recovery process ranged from 21.6 msec to 114.9 msec. CONCLUSIONS: With a dual contact masker (1) off-site listening can be avoided, leading to larger estimates of the width of excitation patterns than in single contact masking, (2) it can be estimated for which stimulation level there is complete overlap of excitation patterns of adjacent electrode contacts, (3) it can be shown that stimulation of nerve fibers in the immediate neighborhood of an electrode contact which were not excited by neighboring electrode contacts is only possible if the probe stimulation amplitude is sufficiently high in comparison with amplitudes on neighboring contacts. PMID- 17086075 TI - The age at which young deaf children receive cochlear implants and their vocabulary and speech-production growth: is there an added value for early implantation? AB - OBJECTIVE: The age at which a child receives a cochlear implant seems to be one of the more important predictors of his or her speech and language outcomes. However, understanding the association between age at implantation and child outcomes is complex because a child's age, length of device use, and age at implantation are highly related. In this study, we investigate whether there is an added value to earlier implantation or whether advantages observed in child outcomes are primarily attributable to longer device use at any given age. DESIGN: Using hierarchical linear modeling, we examined latent-growth curves for 100 children who had received their implants when they were between 1 and 10 yr of age, had used oral communication, and had used their devices for between 1 and 12 yr. Children were divided into four groups based on age at implantation: between 1 and 2.5 yr, between 2.6 and 3.5 yr, between 3.6 and 7 yr, and between 7.1 and 10 yr. RESULTS: Investigation of growth curves and rates of growth over time revealed an additional value for earlier implantation over and above advantages attributable to longer length of use at any given age. Children who had received their implants before the age of 2.5 yr had exhibited early bursts of growth in consonant-production accuracy and vocabulary and also had significantly stronger outcomes compared with age peers who had received their implants at later ages. The magnitude of the early burst diminished systematically with increasing age at implantation and was not observed for children who were older than 7 yr at implantation for consonant-production accuracy or for children who were over 3.5 yr old at implantation for vocabulary. The impact of age at implantation on children's growth curves differed for speech production and vocabulary. CONCLUSIONS: There seems to be a substantial benefit for both speech and vocabulary outcomes when children receive their implant before the age of 2.5 yr. This benefit may combine a burst of growth after implantation with the impact of increased length of use at any given age. The added advantage (i.e., burst of growth) diminishes systematically with increasing age at implantation. PMID- 17086077 TI - Effects of vowel context on the recognition of initial and medial consonants by cochlear implant users. AB - OBJECTIVE: Scores on consonant-recognition tests are widely used as an index of speech-perception ability in cochlear implant (CI) users. The consonant stimuli in these tests are typically presented in the /alpha/ vowel context, even though consonants in conversational speech occur in many other contexts. For this reason, it would be useful to know whether vowel context has any systematic effect on consonant recognition in this population. The purpose of the present study was to compare consonant recognition for the /alpha, i/, and /u/ vowel contexts for consonants presented in both initial (Cv) and medial (vCv) positions. DESIGN: Twenty adult CI users with one of three different implanted devices underwent consonant-confusion testing. Twelve stimulus conditions that differed according to vowel context (/alpha, i, u/), consonant position (Cv, vCv), and talker gender (male, female) were assessed in each subject. RESULTS: Mean percent-correct consonant-recognition scores were slightly (5 to 8%) higher for the /alpha/ and /u/ vowel contexts than for the /i/ vowel context for both initial and medial consonants. This general pattern was observed for both male and female talkers, for subjects with better and poorer average consonant recognition performance, and for subjects using low, medium, and high stimulation rates in their speech processors. In contrast to the mean data, many individual subjects demonstrated large effects of vowel context. For 10 of 20 subjects, consonant-recognition scores varied by 15% or more across vowel contexts in one or more stimulus conditions. Similar to the mean data, these differences generally reflected better performance for the /alpha/ and /u/ vowel contexts than for the /i/ vowel context. An analysis of consonant features showed that overall performance was best for the voicing feature, followed by the manner and place features, and that the place feature showed the strongest effect of vowel context. Vowel-context effects were strongest for the six consonants /d, j, n, k, m/, and /l/. For three of these consonants (/j, n, k/), the back vowels /alpha/ and /u/ produced substantially (30 to 35%) higher mean scores than the front vowel /i/. For each of the remaining three consonants, a unique pattern was observed in which a different single vowel produced substantially higher scores than the others. Several additional consonants (/s, g, w, b/, and /d/) showed strong context effects in either the initial consonant or medial consonant position. Overall, voiceless stop, nasal, and glide-liquid consonants showed the strongest effects of vowel context, whereas the voiceless fricative and voiceless affricate consonants were least affected. Consistent with the feature analysis, a qualitative assessment of phoneme errors for the six key consonants indicated that vowel-context effects stem primarily from changes in the number of place-of articulation errors made in each context. CONCLUSIONS: Vowel context has small but significant effects on consonant-recognition scores for the "average" CI listener, with the back vowels /alpha/ and /u/ producing better performance than the front vowel /i/. In contrast to the average results, however, the effects of vowel context are sizable in some individual subjects. This suggests that it may be beneficial to assess consonant recognition using two vowels, such as /alpha/ and /i/, which produce better and poorer performance, respectively. The present results underscore previous findings that poor transmission of spectral speech cues limits consonant-recognition performance in CI users. Spectral cue transmission may be hindered not only by poor spectral resolution in these listeners but also by the brief duration and dynamic nature of consonant place-of articulation cues. PMID- 17086078 TI - Acoustic change complexes recorded in adult cochlear implant listeners. AB - OBJECTIVES: The objectives of this study were to determine: 1) whether the acoustic change complex (ACC) could be reliably recorded in cochlear implant listeners and, 2) whether different speech sounds evoke distinct ACC patterns. DESIGN: Eight adults wearing the Nucleus-24 cochlear implant (CI) were tested using naturally produced speech tokens /si/ and /i/. Stimuli were tokens from the standardized UCLA version of the Nonsense Syllable Test. Using a repeated measures design, participants were tested and retested within a 3-wk period. RESULTS: Intraclass correlation coefficients for grand mean and individual response waveforms recorded from the syllables /si/ and /i/ ranged from 0.63 to 0.89 from test to retest. Also, ACC latencies signaling the onset of a vowel in /i/ were significantly earlier than those evoked by /si/. CONCLUSIONS: The ACC can be reliably recorded in individuals wearing CI. Furthermore, the naturally produced speech syllables /si/ and /i/ evoke distinct ACC patterns. Because of its good stability and the ease with which it can be recorded in individual CI listeners, the ACC can be evoked using complex signals (such as naturally produced speech syllables) when studying central auditory function in CI listeners. PMID- 17086079 TI - Effects of childhood hearing loss on organization of semantic memory: typicality and relatedness. AB - OBJECTIVE: The purpose of this research was to study how early childhood hearing loss affects development of concepts and categories, aspects of semantic knowledge that allow us to group and make inferences about objects with common properties, such as dogs versus cats. We assessed category typicality and out-of category relatedness effects. The typicality effect refers to performance advantage (faster reaction times, fewer errors) for objects with a higher number of a category's characteristic properties; the out-of-category relatedness effect refers to performance disadvantage (slower reaction times and more errors) for out-of-category objects that share some properties with category members. DESIGN: We applied a new children's speeded category-verification task (vote "yes" if the pictured object is clothing). Stimuli were pictures of typical and atypical category objects (e.g., pants, glove) and related and unrelated out-of-category objects (e.g., necklace, soup). Participants were 30 children with hearing impairment (HI) who were considered successful hearing aid users and who attended regular classes (mainstreamed) with some support services. Ages ranged from 5 to 15 yr (mean = 10 yr 8 mo). Results were related to normative data from . RESULTS: Typical objects consistently showed preferential processing (faster reaction times, fewer errors), and related out-of-category objects consistently showed the converse. Overall, results between HI and normative groups exhibited striking similarity. Variation in speed of classification was influenced primarily by age and age-related competencies, such as vocabulary skill. Audiological status, however, independently influenced performance to a lesser extent, with positive responses becoming faster as degree of hearing loss decreased and negative responses becoming faster as age of identification/amplification/education decreased. There were few errors overall. CONCLUSIONS: The presence of a typicality effect indicates that 1) the structure of conceptual representations for at least one category in the HI group was based on characteristic properties with an uneven distribution among members, and 2) typical objects with a higher number of characteristic properties were more easily accessed and/or retrieved. The presence of a relatedness effect indicates that the structure of representational knowledge in the HI group allowed them to appreciate semantic properties and understand that properties may be shared between categories. Speculations linked the association 1) between positive responses and degree of hearing loss to an increase in the quality, accessibility, and retrievability of conceptual representations with better hearing; and 2) between negative responses and age of identification/amplification/education to an improvement in effortful, postretrieval decision-making proficiencies with more schooling and amplified auditory experience. This research establishes the value of our new approach to assessing the organization of semantic memory in children with HI. PMID- 17086080 TI - A systematic review of the nonacoustic benefits of bone-anchored hearing AIDS. AB - OBJECTIVE: This study completed an evidence-based practice systematic review of the nonacoustic benefits for adult patients receiving bone-anchored hearing aid(s) (BAHAs) relative to other forms of amplification (i.e., none, bone conduction hearing aids [BCHAs], or air-conduction hearing aids [ACHAs]). DESIGN: Systematic review. METHODS: ComDisDome and PubMed were used to perform a comprehensive search for studies that were of at least level III of evidence. Three independent reviewers completed the search, quality assessment, and data extraction. RESULTS: Out of 28 studies, seven studies were identified, three of which were prospective and four of which were retrospective within-subject comparison designs. Nearly all of the studies were of small sample size and/or limited methodological quality, primarily because of logistics (e.g., heterogeneity of subject samples and prior experience with amplification). CONCLUSIONS: The evidence identified in this review is limited regarding the nonacoustic benefits of BAHAs compared with unaided conditions or other types of amplification. Professionals should use caution when counseling patients about the quality of life (QOL) benefits of these devices. Routine use of QOL outcome measures and standardized methods for reporting findings are critical in demonstrating the nonacoustic benefits of BAHAs. PMID- 17086081 TI - Simultaneous bilateral cochlear implantation in adults: a multicenter clinical study. AB - OBJECTIVE: To determine the efficacy of "simultaneous" bilateral cochlear implantation (both implants placed during a single surgical procedure) by comparing bilateral and unilateral implant use in a large number of adult subjects tested at multiple sites. DESIGN: Prospective study of 37 adults with postlinguistic onset of bilateral, severe to profound sensorineural hearing loss. Performance with the bilateral cochlear implants, using the same speech processor type and speech processing strategy, was compared with performance using the left implant alone and the right implant alone. Speech understanding in quiet (CNCs and HINT sentences) and in noise (BKB-SIN Test) were evaluated at several postactivation time intervals, with speech presented at 0 degrees azimuth, and noise at either 0 degrees , 90 degrees right, or 90 degrees left in the horizontal plane. APHAB questionnaire data were collected after each subject underwent a 3-wk "bilateral deprivation" period, during which they wore only the speech processor that produced the best score during unilateral testing, and also after a period of listening again with the bilateral implants. RESULTS: By 6-mo postactivation, a significant advantage for speech understanding in quiet was found in the bilateral listening mode compared with either unilateral listening modes. For speech understanding in noise, the largest and most robust bilateral benefit was when the subject was able to take advantage of the head shadow effect; i.e., results were significantly better for bilateral listening compared with the unilateral condition when the ear opposite to the side of the noise was added to create the bilateral condition. This bilateral benefit was seen on at least one of the two unilateral ear comparisons for nearly all (32/34) subjects. Bilateral benefit was also found for a few subjects in spatial configurations that evaluated binaural redundancy and binaural squelch effects. A subgroup of subjects who had asymmetrical unilateral implant performances were, overall, similar in performance to subjects with symmetrical hearing. The questionnaire data indicated that bilateral users perceive their own performance to be better with bilateral cochlear implants than when using a single device. CONCLUSIONS: Findings with a large patient group are in agreement with previous reports on smaller groups, showing that, overall, bilateral implantation offers the majority of patients advantages when listening in simulated adverse conditions. PMID- 17086082 TI - Does universal newborn hearing screening identify all children with GJB2 (Connexin 26) deafness? Penetrance of GJB2 deafness. AB - OBJECTIVE: Deafness is the most common neurosensory defect at birth, and GJB2 (connexin 26) mutations are the most frequent genetic cause of hearing loss in many populations. The hearing loss caused by GJB2 mutations is usually congenital in onset and moderate to profound in degree. Considerable phenotypic variation has been noted however, including two anecdotal cases of apparent non penetrance at birth. The objective of this study is to document nine additional children with two pathogenic GJB2 mutations who had non penetrance of hearing loss at birth. DESIGN: Subjects were identified through a national repository which includes deaf probands ascertained primarily from the United States through the Annual Survey of Deaf and Hard of Hearing Children and Youth conducted at the Research Institute at Gallaudet University. The hearing of each of these children had been screened at birth using standard audiologic techniques. Parents were interviewed and available medical records were reviewed. Testing for GJB2 mutations was performed by PCR and sequencing of the entire coding exon in all nine individuals. RESULTS: Using parent interviews and medical records, we documented that all nine children passed newborn audiologic hearing screening. The age at which the hearing loss was subsequently identified in these nine children ranged from 12-60 mo. Of these nine children, 3 were compound heterozygotes and six were homozygous for the 35delG mutation in the GJB2 gene. CONCLUSION: These nine cases demonstrate that current newborn hearing screening does not identify all infants with two GJB2 mutations. These cases suggest that the frequency of non penetrance at birth is approximately 3.8% or higher. It is important to consider connexin deafness in any child with recessive nonsyndromic hearing loss as well as simplex cases with no history of other affected family members even when the newborn hearing screening results were within the normal range. PMID- 17086083 TI - Audiogram notches in noise-exposed workers. AB - OBJECTIVES: Diagnostic criteria for noise-induced hearing loss include the audiometric notch, yet no standardized definition exists. This study tested whether objective notch metrics could match the clinical judgments of an expert panel. DESIGN: A panel of occupational physicians, otolaryngologists, and audiologists reviewed audiograms of noise-exposed workers. In a two-sample process, the panel judged whether a notch was present and whether hearing loss had progressed in a notch pattern. Quantitative notch metrics were compared against expert decisions. RESULTS: At least five of six experts agreed about notch identification in 71 and 72% of the cases in the two samples, and agreement about notch progression was 61 and 67%. Notch depth and professional specialty appeared to affect notch judgments. Despite this variability, a notch metric showed excellent agreement with expert notch consensus in each sample (94.7 and 96.6%; kappa = 0.88 and 0.92). CONCLUSIONS: Audiogram notch metrics can agree with expert clinical consensus and assist in the surveillance of noise-exposed workers. PMID- 17086084 TI - Comparison of auditory steady-state responses and tone-burst auditory brainstem responses in normal babies. AB - OBJECTIVE: To follow the development of tone-burst auditory brainstem response (TB-ABR) and auditory steady-state response (ASSR) thresholds in a group of normal babies through the first 6 wk of life. DESIGN: This longitudinal study involved assessment at four data-collection points. TB-ABR and ASSR thresholds to 500-Hz and 4-kHz stimuli were established in 17 full-term subjects at 0, 2, 4, and 6 wk of age. Stimulus-modulation rates for ASSR assessment were 74 Hz (for 500-Hz tones) and 95 Hz (for 4-kHz tones). TB-ABR responses were recorded to stimuli presented at 39.1 Hz. RESULTS: Mean ASSR thresholds (calibrated in dBHL) at 500 Hz ranged from 44.4 to 39.7 dB HL across the recording period, and at 4 kHz they ranged from 37.9 to 32.1 dB HL. TB-ABR thresholds (calibrated in dBnHL) were significantly lower, ranging from 36.8 to 36.2 dB nHL at 500 Hz and from 16.5 to 15.9 dB nHL at 4 kHz. However, when the stimuli used for each test were calibrated in the same units (peak equivalent dB SPL), the results were similar. That is, the differences between the two techniques were only an artifact of the calibration. ASSR thresholds were more variable than TB-ABR, particularly at the neonatal measurement point. Within-subject changes across the test period were observed for ASSR thresholds but not for TB-ABR. CONCLUSIONS: The longitudinal findings presented in this study suggest that for normal neonates, the TB-ABR technique may offer a more reliable basis for prediction of hearing levels than ASSR assessment. This is not because TB-ABR thresholds (calibrated in dBnHL) are lower, but because the response is less affected by maturational development in the first weeks of life and is less variable across subjects. PMID- 17086085 TI - Speech recognition for unilateral and bilateral cochlear implant modes in the presence of uncorrelated noise sources. AB - OBJECTIVE: The purpose of the current investigation was to compare speech recognition in noise for bilateral and unilateral modes within postlingually deafened, adult bilateral cochlear implant recipients. In addition, it was of interest to evaluate the time course of the bilateral speech-recognition advantage and the effect of changing signal-to-noise ratio (SNR) on the magnitude of the bilateral advantage. DESIGN: In the first experiment, 16 postlingually deafened adults who were bilaterally implanted with the MED-EL C40+ cochlear device were evaluated in unilateral left, unilateral right, and bilateral conditions 4 to 7 mo after activation. Speech recognition in the presence of five spatially separated, uncorrelated noise sources was evaluated using both a single fixed SNR of +10 dB and an adaptive-SNR method. In a follow-up study, a subset of 10 participants was re-evaluated using an identical fixed-SNR method 12 to 17 mo after activation to examine the time course of speech-recognition performance in both unilateral and bilateral modes at a single SNR. A third study was performed with a subset of six participants to examine performance over a range of SNRs. In this study, speech recognition was measured 12 to 17 mo after activation in quiet and at +5, +10, +15, and +20 dB SNRs using the same five uncorrelated noise sources. RESULTS: The speech-recognition data revealed a significant bilateral advantage of 3.3 dB using the adaptive-SNR method. A significant bilateral advantage of 9% was also measured using a fixed +10 dB SNR. Results from the second study revealed that experience resulted in a significant (11 to 20%) increase in speech-recognition-in-noise performance for both unilateral and bilateral modes; however, the magnitude of the bilateral advantage was not affected by experience. Results from the third study revealed the largest bilateral advantage at the poorest SNR evaluated. In addition, performance in quiet was significantly better than that measured in the presence of noise, even at the +20 dB SNR. CONCLUSIONS: The results of these experiments support a small but significant bilateral speech-recognition-in-noise advantage for cochlear implant recipients in an environment with multiple noise sources. This advantage is presumed to be attributable to the combined effects of binaural squelch and diotic summation. Although experience generally improved speech-recognition-in noise performance in both unilateral and bilateral modes, a consistent bilateral advantage (approximately 10%) was measured at 4 to 7 mo and at 12 to 17 mo postactivation. PMID- 17086086 TI - Wideband reflectance norms for Caucasian and Chinese young adults. AB - OBJECTIVE: This study examined differences between the middle ears of two ethnic groups, Caucasian and Chinese young adults with normal hearing, using a new middle-ear measurement technique, wideband energy reflectance. The goal of this study was to determine whether the Chinese group had different middle-ear transmission properties than the Caucasian group. DESIGN: There were 126 subjects (237 ears) between the ages of 18 and 32 yr, with 62 subjects in the Caucasian group and 64 subjects in the Chinese group. Wideband energy reflectance data were gathered using Mimosa Acoustics (RMS system version 4.0.4.4) wideband reflectance (WBR) equipment. RESULTS: The Chinese group had significantly lower wideband energy reflectance than their Caucasian counterparts at higher frequencies; however, the Caucasian group had significantly lower energy reflectance at lower frequencies than the Chinese group. The Chinese group also had significantly lower admittance magnitude than the Caucasian group at lower frequencies. Because body size indices are more comparable between Caucasian females and Chinese males, the effect of body size could be potentially adjusted for by comparing the Caucasian female subjects with the Chinese male subjects. The differences observed between the Caucasian and the Chinese groups were no longer significant when the Caucasian female subjects were compared with the Chinese male subjects. Applying the Caucasian norms to four Caucasian adults with surgically confirmed otosclerosis resulted in an improved hit rate compared with the combined Caucasian and Chinese norms and the Chinese-only norms. CONCLUSION: Body size may play a role in the observed differences between the Caucasian and Chinese groups. The findings of this study suggest that further research is needed to investigate the effects of body size on wideband energy reflectance. It should be noted that factors other than body size may contribute to the observed differences. Chinese individuals may simply have different middle-ear characteristics than Caucasian individuals, which could affect WBR. In the meantime, overall test performance may be improved by using a more homogeneous norm when evaluating the middle ear of Caucasian or Chinese individuals with WBR. PMID- 17086087 TI - Comparing cochlear implant users' speech performance with processor fittings based on conventionally determined T and C levels or on compound action potential thresholds and live-voice speech in a prospective balanced crossover study. AB - OBJECTIVE: The objective of the present study is to improve the efficiency of the fitting procedure of cochlear implant processors by making use of measurements of the electrically evoked compound actio potential (ECAP) and live-voice speech. DESIGN: In a randomised prospective cross-over design we compare speech performance of eighteen adult subjects when following the conventional fitting procedure to a procedure in which we use the profile of the ECAP threshold levels across the full electrode array measured intra-operatively. The overall level of the profile is shifted (by an equal amount of current units per electrode) until we find the threshold for live speech (new T levels) and the loudness comfort level (new C levels). Each fitting procedure is tested for 6 wk. Speech performance is measured in quiet and in noise every other week. RESULTS: The results show little difference between the scores (Dutch CVC words) for the conventional fitting procedure and the ECAP based fitting, although the T and C levels may differ markedly. CONCLUSION: The new fitting procedure is much faster and easier in the initial phase. Further improvement of performance may be obtained in a later stage of the fitting procedure by changing some individual electrodes on the basis of subjective responses. PMID- 17086090 TI - The effect of angiotensin II receptor blockade on an end-stage renal failure model of type 2 diabetes. AB - The effect of olmesartan medoxomil (OLM), an angiotensin II receptor blocker (ARB), on advanced nephropathy and mortality was evaluated in Zucker Diabetic Fatty (ZDF) rats, a type 2 diabetes model. OLM was administered from 36 weeks of age, when the animals developed advanced proteinuria. OLM effectively suppressed the progression of proteinuria. The ZDF rats started to die at 50 weeks of age, which was accompanied by abrupt increase in blood urea nitrogen, suggesting that the cause of death was renal insufficiency. OLM suppressed increases in blood urea nitrogen and increased the survival rate of the ZDF rats. The histological examination revealed that the renal damage was ameliorated by OLM. The macrophage infiltration and monocyte chemoattractant protein-1 (MCP-1) expression was increased in the glomeruli and tubulointerstitium of the ZDF rat kidneys, and the increase was lessened by OLM. In a separate study, albumin increased MCP-1 release from cultured tubular epithelial cells. These results suggest that protein leakage from the glomeruli stimulates MCP-1 production in tubular cells and that MCP-1 released into the interstitial space induces macrophage infiltration and inflammation. It is conceivable that the beneficial actions of ARB on diabetic nephropathy are, at least in part, due to decrease of proteinuria and the subsequent reduction of inflammatory changes in tubular cells. PMID- 17086091 TI - Neutrophil superoxide anion generation during atorvastatin and fluvastatin therapy used in coronary heart disease primary prevention. AB - Neutrophil superoxide anion generation was measured during atorvastatin and fluvastatin therapy in patients with coronary heart disease (CHD) risk. The patients were randomly allotted into three groups. The atorvastatin group comprised 17 patients who were administered the drug orally 10 mg a day at bed time. The fluvastatin group consisted of 18 patients on an oral dose of 40 mg once daily at bed time. The control group comprised 12 healthy subjects with no drug administration. Blood samples were collected from cubital vein before and after 6-week therapy with these drugs and once in the control group. Neutrophil superoxide anion generation in whole blood without and with opsonized zymosan (OZ) stimulation was determined using superoxide dismutase from bovine erythrocytes. In the atorvastatin group, statistically significant (P < 0.05) decrease in superoxide anion generation by nonstimulated and OZ-stimulated neutrophils was observed after 6 weeks of therapy. In fluvastatin group, no changes in neutrophil superoxide anion generation were observed after the 6-week treatment period. Our study has shown an additional nonlipid mechanism of atorvastatin used in CHD primary prevention. PMID- 17086092 TI - Preconditioning effects of levosimendan in a rabbit cardiac ischemia-reperfusion model. AB - The preconditioning effects of levosimendan were investigated on ischemia reperfusion induced morphological and functional cardiac damage. Langendorff perfused rabbit hearts were reserved as controls or subjected either to global myocardial ischemic preconditioning or to perfusion with levosimendan (0.1 micromol/l) for two 5-minute cycles. After a washout period, all hearts were then subjected to 30 minutes of global ischemia and 120 minutes of drug-free reperfusion. Intraventricular pressure and coronary flow were measured, and infarct size determined after nitroblue-tetrazolium staining on completion of the experiments. Levosimendan pretreatment resulted in a significantly smaller elevation from the preischemic level in left ventricular end-diastolic pressure during reperfusion (37 +/- 17 mm Hg) compared with controls (56 +/- 14 mm Hg) and ischemia-preconditioned hearts (53 +/- 34 mm Hg). The left ventricular developed pressure-representing the functional recovery of the heart after ischemia-that was significantly improved by levosimendan pretreatment (38 +/- 6% vs 16 +/- 5% in controls, P < 0.05). In addition, contractility and relaxability parameters (+dP/dt and -dP/dt, respectively) were better preserved in the levosimendan hearts. The volume of infarcted myocardium after global ischemia-reperfusion was significantly (P < 0.05) decreased by both ischemic preconditioning (38 +/- 2%) or levosimendan pretreatment (45 +/- 2%) versus controls (52 +/- 2%). The results of this study suggest that levosimendan pretreatment is capable of decreasing infarct size in an ischemia-reperfusion model and improving recovery of cardiac function following ex vivo global ischemia. PMID- 17086093 TI - Normal aging impairs upregulation of the beta-adrenergic but not the alpha adrenergic response: aging and adrenergic upregulation. AB - OBJECTIVES: To determine if centrally reducing sympathetic tone with clonidine will reverse the downregulation in the alpha-adrenergic (alphaAR) and beta adrenergic (betaAR) responses seen with normal aging. METHODS: Twelve rigorously screened young adult (mean age, 26 years) and 15 older adult (mean age, 69 years) subjects were studied before and after using the clonidine patch (TTS-2) for 2 weeks. betaAR (isoproterenol at 35 ng/kg/min) and alphaAR (phenylephrine at 1.0 microg/kg/min) were assessed using radionuclide measures of end diastolic, end systolic, and stroke volume indices, cardiac index, and ejection fraction. RESULTS: Clonidine reduced resting plasma norepinephrine and this reduction was greater in older subjects (-47 +/- 3 versus -26 +/- 6%, P = 0.001). After 2 weeks of clonidine patch, upregulation of the betaAR was significantly higher in young subjects for heart rate (+10.7 +/- 1.5 versus +4.6 +/- 1.5 bpm; P = 0.01). There was no significant age-associated difference in the upregulation of the alphaAR with clonidine for systolic, diastolic, and mean blood pressure or systemic vascular resistance. CONCLUSIONS: With aging, there is an impaired resensitization of the chronotropic betaAR response with central sympathetic downregulation that is not seen with the alphaAR. PMID- 17086094 TI - A therapeutic dose of the lipophilic statin pitavastatin enhances oxidant-induced apoptosis in human vascular smooth muscle cells. AB - We examined effects of a physiologic concentration of pitavastatin (0.01 micromol/L) on oxidant-induced apoptosis in cultured human vascular smooth muscle cells (VSMCs). Apoptosis was induced in VSMCs by hydrogen peroxide (H2O2, 300 micromol/L), as evidenced by in situ nick end-labeling and scanning electron microscopy. This apoptotic response was accompanied by increased activation of mitogen-activated protein kinases (MAPKs--ie, increases in the phosphorylated forms of extracellular signal-regulated kinase (p-ERK), c-Jun N-terminal kinase (p-JNK), and p38 MAPK (p-p38 MAPK). Although pitavastatin alone did not induce VSMC death, pretreatment with pitavastatin significantly enhanced H2O2-induced apoptosis and prolonged activation of JNK and p38 MAPK (for up to 24 h) but not ERK. Expression of MAPK phosphatase-1 (MKP-1) also was upregulated by H2O2, but this was not affected by pitavastatin. The apoptosis accelerating effect was observed also in simvastatin but not in pravastatin. Treating VSMCs with mevalonate, farnesyl pyrophosphate, or geranylgeranyl pyrophosphate completely blocked the statin-induced enhancement of VSMC apoptosis, suggesting that protein prenylation is critically involved. It thus appears that pitavastatin enhances H2O2-induced VSMC apoptosis, at least in part, via increases in MAPK activation and protein prenylation, but independently of MKP-1 expression, which consequently results in reduction of VSMC population. PMID- 17086095 TI - Neurohumoral changes in patients with left ventricular dysfunction following acute myocardial infarction and the effect of nitrate therapy: a randomized, double-blind, placebo-controlled long-term study. AB - BACKGROUND: Several neurohumoral mechanisms involved in cardiovascular regulation are activated in the failing heart, but only limited information is available regarding the influence of long-term nitrate therapy. MATERIALS AND METHODS: This was a double-blind, randomized comparison of isosorbide-5-mononitrate (IS-5-MN), 60 mg given orally, once daily for 11 months to patients (n = 47) with left ventricular (LV) dysfunction following acute myocardial infarction (AMI). Forty five patients received placebo. All patients received ramipril.Plasma natriuretic peptides (atrial [ANP] and brain [BNP] natriuretic peptide), epinephrine, norepinephrine (NEPI), antidiuretic hormone, aldosterone (Aldo), renin activity (PRA), substance P, neuropeptide Y-like immunoreactivity, calcitonin gene-related peptide, and vasoactive intestinal peptide were measured at baseline and at the end of the treatment period. Clinical, echocardiographic, and hemodynamic data were also obtained. RESULTS AND CONCLUSIONS: Chronic nitrate therapy does not significantly affect the neurohumoral status in patients with LV dysfunction after AMI, apart from a decrease in ANP. Some hormones are more closely associated with diastolic dysfunction/increased volume load (ANP and BNP) and others are more closely associated with systolic dysfunction (PRA, NEPI, Aldo). There is a temporal dissociation of these 2 groups of hormones 1 year post infarction: ANP and BNP decrease, whereas NEPI and Aldo show a slight increase. BNP levels do not reflect all important pathophysiologic mechanisms in heart failure. Consequently, the use of other neurohormonal factors than BNP for monitoring of heart failure therapy should be explored. PMID- 17086096 TI - Potent potentiating diuretic effects of prednisone in congestive heart failure. AB - Animal studies showed glucocorticoids could specifically dilate renal vasculature, regulate synthesis and release of atrial natriuretic peptide (ANP), upregulate ANP receptors on vascular endothelial cells, and thus have potent potentiating diuresis and natriuresis effects in animal studies; however, their diuretic efficacy in humans is yet to be known. Therefore, we designed this randomized, double- blind, placebo-controlled, clinical study to determine the diuretic efficacy of prednisone, a glucocorticoid, in patients with congestive heart failure (CHF). METHODS: Twenty clinically stable patients with CHF without overt fluid retention were randomized to a prednisone group or placebo group. Prednisone (1 mg/kg/day with a maximum dose of 60 mg/day) was added to standard care for 7 days, leaving other medications unchanged. Variables included urine volume and electrolytes, serum electrolytes, and change from baseline in serum creatinine. RESULTS: Adding prednisone resulted in striking diuresis and natriuresis with time. As compared with the placebo group, the maximum of mean daily urine volume was 810.5 mL larger than those in the placebo group (95% confidence intervals [CI] 276.25 to 1344.86, P < 0.05). The maximum mean daily sodium excretion was 123.8 mmol higher than those patients given placebo (95% CI 11.4 to 236.2, P < 0.05). The placebo-corrected effect on change from baseline in serum creatinine was -19.5 mumol/L (95% CI -7.4 to -31.6, P < 0.01), favoring prednisone. CONCLUSIONS: This pilot study showed that prednisone had potent potentiating diuretic effects in patients with heart failure and might improve renal function in the same time. Further prospective randomized clinical studies are warranted to determine the preferable dose and its efficacy in decompensated congestive heart failure. PMID- 17086097 TI - Antioxidants suppress plasma levels of lectinlike oxidized low-density lipoprotein receptor-ligands and reduce atherosclerosis in watanabe heritable hyperlipidemic rabbits. AB - Oxidative modification of low-density lipoprotein (LDL) has been implicated in the pathogenesis of atherosclerosis. In this study, we investigated the effects of antioxidants including probucol, vitamin E, and fluvastatin, an HMG-CoA (hydroxy-3-methylglutaryl coenzyme A) reductase inhibitor with antioxidative property, on plasma levels of oxidized LDL (OxLDL) during the progression of atherosclerosis in Watanabe heritable hyperlipidemic (WHHL) rabbits. OxLDL were measured as ligand for lectin-like OxLDL receptor-1 (LOX-1). LOX-1-ligand was higher in WHHL rabbits than in control rabbits as early as 2 months of age and was sustained throughout the experimental period. Supplementation of probucol (1%) and vitamin E (0.5%) to the diet reduced LOX-1-ligand but had little effect on total cholesterol (T-CHO). Fluvastatin (0.03%) significantly reduced both LOX 1-ligand and T-CHO. The extent of reduction in T-CHO was less prominent than in the case of LOX-1-ligand. All of the agents reduced the atherosclerotic lesion area and lipid contents of aortic arches. These parallel results indicate that oxidatively modified LDL elevated in the early stages of atherogenesis is of functional importance in the progression of the disease and can be suppressed by antioxidant treatment. Furthermore, fluvastatin may reduce the evolution of atherosclerosis, not only by lowering plasma cholesterol but also by reducing oxidative modification of LDL. PMID- 17086098 TI - Stent-based delivery of antisense oligodeoxynucleotides targeted to the PDGF A chain decreases in-stent restenosis of the coronary artery. AB - BACKGROUND: Although the use of drug-eluting stents (DESs) has been shown to limit neointima hyperplasia, currently available DESs may adversely affect reendothelialization, possibly precipitating cardiac events. We evaluated the effect of an antisense oligodeoxynucleotide (ODN) targeted to the platelet derived growth factor (PDGF) A-chain on in-stent restenosis in pig coronary artery. METHODS: A bare metal stent coated with phosphorothioate-linked antisense ODN or nonsense ODN, or a bare metal stent without ODN (control), was implanted in the mid segment of the left anterior descending artery (LAD). Twenty-eight days after implantation, angiography and intravascular ultrasound (IVUS) were performed, the LAD was removed, and stenosis was evaluated pathologically. RESULTS: Volumetric stenosis ratios were 64 +/- 11.9, 44 +/- 3.4, and 26 +/- 3.8% in coronary arteries implanted with control, nonsense ODN-coated, and antisense ODN-coated stents, respectively. In angioscopic findings, the lumen surface was smooth in the stented segments in all groups. Struts of antisense ODN-coated stents were observed embedded in the neointima, whereas embedding was not observed in nonsense ODN-coated stents or control stents, indicating a decrease in hyperplasia in response to antisense ODN treatment. Pathologic findings showed 77 +/- 5.8, 68 +/- 12.2, and 38 +/- 5.3% stenosis in coronary arteries implanted with control stents, nonsense ODN-coated stents, and antisense ODN-coated stents, respectively. A continuous lining of endothelial cells was observed along the lumen of coronary arteries implanted with antisense ODN-coated stents. CONCLUSIONS: Stent-based delivery of an antisense ODN targeted to the PDGF A chain effectively inhibits neointima formation after stent implantation in pig coronary artery by suppressing VSMC hyperplasia and preserving endothelialization. Antisense-ODNs may provide a therapy for in-stent restenosis of the coronary artery. PMID- 17086099 TI - The application of microarray technology to neuropathology: cutting edge tool with clinical diagnostics potential or too much information? AB - Microarray technology is a tremendously powerful method for simultaneously monitoring the expression of thousands of species of nucleic acids, usually cellular mRNA, producing a high-resolution representation of the genes encoded or expressed in a cell. As such, microarray technology has great potential for impacting research and clinical approaches to treatment. However, this complex technology has been challenging to apply as a result of difficulties discerning biologic variation from technologic issues, therefore slowing the application of the technology to human diagnostics. Nevertheless, significant advances in microarray technology, improvements that avoid potential pitfalls, and a wider spectrum of application are making this technology easier to apply. Indeed, microarray technology has provided valuable insights into mechanisms involving gene regulation and expression in Alzheimer disease, and it remains a powerful tool to identify biomarkers for disease diagnosis. Ultimately, the most robust markers will enable the application of more specific treatments particular to disease stages or subcategories. Currently, no widely applicable molecular test is available to identify those at risk for developing Alzheimer disease or those who have early markers of pathology but show discernible cognitive impairment. The progression of this technology will lead to earlier detection of the disease through enhanced understanding of disease onset and progression. PMID- 17086100 TI - Abeta species removal after abeta42 immunization. AB - Neuropathologic examination of 3 patients with Alzheimer disease in the Elan Pharmaceuticals trial using antibodies specific for different Abeta species showed in one case, 4 months after the immunization, evidence of a stage of active plaque clearance with "moth-eaten" plaques and abundant Abeta phagocytosis by microglia. At 1 to 2 years after immunization, 2 cases showed extensive areas cleared of plaques (69% and 86% of the temporal cortex was plaque-free). Cortex cleared of plaques in all 3 cases had a characteristic constellation of features, including a very low plaque burden, sparse residual dense plaque cores, and phagocytosed Abeta within microglia. There was resolution of tau-containing dystrophic neurites, although other features of tau pathology (tangles and neuropil threads) remained and cerebral amyloid angiopathy persisted. Although most antibodies generated by Abeta42 immunization in humans bind the intact N terminus, immunohistochemistry with specific antibodies showed clearance of all major species of Abeta (Abeta40, Abeta42, and N-terminus truncated Abeta). Abeta immunotherapy can clear all Abeta species from the cortex. However, if it is to be used for treatment of established Alzheimer disease, then the residual tau pathology and cerebral amyloid angiopathy require further study. PMID- 17086101 TI - Genomic analysis of pilocytic astrocytomas at 0.97 Mb resolution shows an increasing tendency toward chromosomal copy number change with age. AB - Brain tumors are the most common solid tumors of childhood, accounting for over 20% of cancers in children under 15 years of age. Pilocytic astrocytomas (PAs), World Health Organization grade I, are one of the most frequently occurring childhood brain tumors, yet little is known about genetic changes characterizing this entity. We have used microarray comparative genomic hybridization at 0.97 Mb resolution to study a series of PAs (n = 44). No copy number abnormality was seen in 64% of cases at this resolution. However, whole chromosomal gain (median 5 chromosomes affected) occurred in 32% of tumors. The most frequently affected chromosomes were 5 and 7 (11 of 44 cases each) followed by 6, 11, 15, and 20 (greater than 10% of cases each). Findings were confirmed by fluorescence in situ hybridization and microsatellite analysis in a subset of tumors. Chromosomal gain was significantly more frequent in PAs from patients over 15 years old (p = 0.03, Fisher exact test). The number of chromosomes involved was also significantly greater in the older group (p = 0.02, Mann-Whitney U test). One case (2%) showed a region of gain on chromosome 3 and one (2%) a deletion on 6q as their sole abnormalities. This is the first genomewide study to show this nonrandom pattern of genetic alteration in pilocytic astrocytomas. PMID- 17086102 TI - The human prion protein residue 129 polymorphism lies within a cluster of epitopes for T cell recognition. AB - T cell immune responses to central nervous system-derived and other self-antigens are commonly described in both healthy and autoimmune individuals. However, in the case of the human prion protein (PrP), it has been argued that immunologic tolerance is uncommonly robust. Although development of an effective vaccine for prion disease requires breaking of tolerance to PrP, the extent of immune tolerance to PrP and the identity of immunodominant regions of the protein have not previously been determined in humans. We analyzed PrP T cell epitopes both by using a predictive algorithm and by measuring functional immune responses from healthy donors. Interestingly, clusters of epitopes were focused around the area of the polymorphic residue 129, previously identified as an indicator of susceptibility to prion disease, and in the C-terminal region. Moreover, responses were seen to PrP peptide 121-134 containing methionine at position 129, whereas PrP 121-134 [129V] was not immunogenic. The residue 129 polymorphism was also associated with distinct patterns of cytokine response: PrP 128-141 [129M] inducing IL-4 and IL-6 production, which was not seen in response to PrP 128-141 [129V]. Our data suggest that the immunogenic regions of human PrP lie between residue 107 and the C-terminus and that, like with many other central nervous system antigens, healthy individuals carry responses to PrP within the T cell repertoire and yet do not experience deleterious autoimmune reactions. PMID- 17086103 TI - Prognostic implications of atypical histologic features in choroid plexus papilloma. AB - The prognostic significance of atypical histologic features in choroid plexus tumors remains uncertain. Therefore, a series of 164 choroid plexus tumors was evaluated for the presence of atypical histologic features, including mitotic activity, increased cellularity, nuclear pleomorphism, blurring of papillary growth pattern, and necrosis. The impact of histopathologic and clinical features on the probability of recurrence and survival was investigated. Twenty-four tumors displaying frank signs of malignancy were diagnosed as choroid plexus carcinoma according to World Health Organization criteria. Of 124 choroid plexus papillomas that had not received adjuvant treatment, 46 tumors (37%) displayed at least one atypical feature, including increased cellularity (n = 25 [20%]), mitotic activity (> or =2 mitoses per 10 high-power fields; n = 19 [15%]), nuclear pleomorphism (n = 16 [13%]), solid growth (n = 15 [12%]), and necrosis (n = 5 [4%]). Only one tumor-related death, but 10 recurrences, were observed on a mean observation time of 58 months. On univariate analysis, incomplete surgical resection (p = 0.03) and mitotic activity (p < 0.001) were the only clinicopathologic factors associated with recurrence. Using a multivariate model, an independent effect of mitotic activity on the probability of recurrence could be confirmed (p = 0.001). Because mitotic activity is the sole atypical histologic feature independently associated with recurrence, we propose to define atypical choroid plexus papilloma by mitotic activity (> or =2 mitoses per 10 high-power fields) corresponding to World Health Organization grade II, thus adjoining other intermediate tumor entities associated with increased mitotic activity such as atypical meningioma. Close follow up of patients harboring atypical choroid plexus papillomas may be warranted. PMID- 17086104 TI - Effect of selective cholinergic denervation on the serotonergic system: implications for learning and memory. AB - The cholinergic system has been widely implicated in cognitive processes and cholinergic loss is a classical hallmark in Alzheimer disease. Increasing evidence supports a role of the serotonergic system in cognition, possibly through a modulation of cholinergic activity. We compared selective cholinergic denervation by administration of the immunotoxin 192 IgG-saporin in the nucleus basalis of Meynert (NBM) with intracerebroventricular (ICV) lesions of the basal forebrain in male rats 7 days after lesioning. NBM lesions induced significant changes in cholinergic markers in the frontal cortex, whereas ICV lesions produced significant decreases in cholinergic markers both in the frontal cortex and hippocampus. Only ICV lesions lead to memory impairments in passive avoidance and Morris water maze tasks. Both models lead to reductions of serotonin levels in the frontal cortex. Similar changes in 5-hydroxytriptophan levels were observed, suggesting a downregulation of the rate-limiting enzyme for the synthesis of serotonin along with the cholinergic deficit. Neither 5-HT1A nor 5 HT1B receptors seem to mediate this process. These data imply that the serotonergic system in the frontal cortex can compensate for diminished cholinergic function and support the investigation of the serotonergic system as a therapeutic target to treat Alzheimer disease. PMID- 17086105 TI - Plaque-derived oxidative stress mediates distorted neurite trajectories in the Alzheimer mouse model. AB - Alzheimer disease (AD) is characterized both by senile plaques and neurodegeneration, although the details of the relationship between the 2 are not well understood. We postulated that oxidative stress resulting from senile plaques may mediate plaques' effects on local neuronal processes. Using multiphoton microscopy, we directly demonstrate the generation of reactive oxygen species by senile plaques. After screening of several natural antioxidants ex vivo, we assessed in vivo the effect of 2 orally administered antioxidants in APPswe/PS1d9 transgenic mice. Both Ginkgo biloba extract and vitamin E reduced the oxidative stress resulting from senile plaques in vivo as monitored with intracranial imaging. Both treatments also lead to a progressive reversal of the structural changes in dystrophic neurites associated with senile plaques. These results suggest a causal relationship between plaque-associated oxidative stress and neuritic alterations and demonstrate for the first time that the focal neurotoxicity associated with the senile plaques of AD is partially reversible with antioxidant therapies. The quantitative ex vivo screen combined with in vivo monitoring of efficacy should lead to more effective clinical therapies for the prevention of oxidative stress and neurotoxicity in AD. PMID- 17086106 TI - Upregulated expression of purinergic P2X(7) receptor in Alzheimer disease and amyloid-beta peptide-treated microglia and in peptide-injected rat hippocampus. AB - The expression of the purinergic receptor subtype P2X(7)R, a nonselective cationic channel activated by high levels of adenosine triphosphate (ATP), has been studied in adult microglia obtained from Alzheimer disease (AD) and nondemented (ND) brains, in fetal human microglia exposed to Abeta(1-42) peptide and in vivo in Abeta(1-42)-injected rat hippocampus. Semiquantitative reverse transcriptase-polymerase chain reaction showed enhanced expression (increase of 70%) of P2X(7)R in AD microglia compared with ND cells (analysis of 6 AD and 8 ND cases). Immunohistochemical analysis showed prominent P2X(7)R expression in association with Abeta plaques and localized to HLA-DR-immunoreactive microglia. In cultured fetal human microglia, cells exposed to Abeta(1-42) (5 microM for 18 hours) had significantly elevated levels of P2X(7)R (by 106%) compared with untreated cells. Amplitudes of Ca(2+) responses in these cells, induced by the selective P2X(7)R agonist BzATP, were increased by 145% with Abeta(1-42) pretreatment relative to control (no peptide pretreatment) and were largely blocked if the P2X(7)R inhibitor-oxidized ATP (oxATP) was added with peptide in pretreatment solution. In vivo, double immunostaining analysis showed considerable P2X(7)R colocalized with microglia after injection of Abeta(1-42) (1 nmol) into rat hippocampus. The overall results suggest roles of P2X(7)R in mediating microglial purinergic inflammatory responses in AD brain. PMID- 17086111 TI - [Organ transplants and organ shortages: two sides of success]. PMID- 17086112 TI - [Permanent tattoos: new complications for the 21st century]. PMID- 17086113 TI - [Organ and tissue shortage in France]. AB - INTRODUCTION: Despite an increase in the number of brain deaths over the past 10 years, France faces a shortage of organs for transplantation. A high percentage of families oppose organ donation. Over the past decade, the mean age of organ donors has risen by 10 years. METHODS: Specific provisions of the public health code have regulated transplantation activity since 1994. Implementation of the Transplant Plan in 2000 led to the investment of substantial financial and human resources in this area. The data reported here come from the 2004 report of the French Transplantation Agency, now the Biomedicine Agency, and from its database. RESULTS: Data for 2004 show more than 2500 listed donors, with at least one organ taken from 1291 and more than 4000 organ transplants. The donors' mean age was 47 years. Cerebrovascular diseases accounted for most of the deaths; traffic accidents, which have decreased substantially in recent years, caused only 16%. The number of people with brain death reported in France is difficult to measure but we estimate the figure to be 3500. Refusal rates remain high and are an obstacle to increasing transplants. DISCUSSION: We need to appeal to live donors. We are also setting up pilot programs to use organs from non-heart-beating donors. With these new initiatives we hope to reach rates equal to those in Spain, the European leader in organ transplants. PMID- 17086114 TI - [Knowledge of legal aspects of organ donation by live donors. Staff survey at a university hospital center]. AB - OBJECTIVE: Law no. 2004-800 dated August 6, 2004, establishes new legal measures to facilitate live organ donation, in view of the shortage of organs for transplantation. We sought to assess the knowledge of this statute among hospital personnel. METHOD: We sent an e-mail questionnaire to 1015 persons working at Amiens university hospital center. It contained eleven closed questions (yes/no/don't know) about permissible live organ and tissue donation and two questions requesting personal opinions about live donation. We compared responses according to profession and department. RESULTS: Questionnaires were returned by 245 respondents. The questions for which more than 70% provided correct answers concerned the fundamental principles of bioethics (consent revocable at any time, donations by adults, donations between spouses, bone marrow donations by minors). Fewer than half the respondents knew the correct answers about the new legal measures adopted in the statute (extension of the circle of potential donors, possible exceptions for donations by minors and incompetent adults). Knowledge of the law concerning assisted reproduction techniques was much poorer: fewer than 30% knew that sperm or oocyte donation requires spousal consent. Three quarters of the respondents have discussed their wishes with their family; 65% would provide a live kidney donation, although some only for a family member. In all 31% have organ donor cards. DISCUSSION: Donations by live donors raise psychological as well as ethical and legal issues. This survey increased the awareness of hospital staff, provided them with information about the relevant law and, we hope, stimulated their thinking about the topic of live donations. PMID- 17086115 TI - [Infiltrating breast cancer in women younger than 25 years: 13 cases]. AB - OBJECTIVE: To study the prognosis of breast cancer in woman younger than 25 years and to compare it with that of other age groups to identify prognostic and histologic factors specific to this group. METHOD: Retrospective study of all cases of infiltrating ductal carcinoma treated at our hospital from January 1977 through July 2005, examining clinical, histologic, and treatment variables as well as 5-year overall survival and 5-years disease-free survival rates. RESULTS: The study included 13 women younger than 25 years at diagnosis. Their average age at diagnosis was 23.3 years (CI=1 year). Time from initial signs of disease until diagnosis averaged 6.6 months (CI=2.5). Clinically, the average tumor size was 28.78 mm (CI=6.06), with 46% classified as T1, 31% as T2 and 23% as T4d. We found 92.3% to be invasive ductal carcinoma, with 30% including an in-situ component; 53.8% were SBR grade 3 and 23% included axillary node invasion. Hormone receptors were present in 61.5% of tumors. During the follow-up period, we observed two deaths (with a 5-year overall survival rate, however, of 91%) and 6 recurrences (5-year disease-free survival: 66.5%). CONCLUSION: Prognosis appears unfavorable among young women (younger than 40 years) with breast cancer. In our series, neither prognosis nor clinical or histologic characteristics differed in the subgroup of very young women (younger than 26 years). PMID- 17086116 TI - [BNP tests in the emergency department to diagnose congestive heart failure]. AB - OBJECTIVES: B-type natriuretic peptide (BNP) testing is a useful tool for diagnosis of congestive heart failure (CHF). In our institution only physicians in the lung disease, cardiology, and intensive care departments were authorized to order BNP testing. Our study sought to determine the validity of this strategy. METHODS: In this one-year prospective study, BNP testing was performed only in the lung disease and cardiology departments. The following details were recorded for each patient with a BNP assay: demographic information, clinical symptoms, physical examination, laboratory reports, chest radiography and ECG findings, and initial diagnosis (including in particular suspicion of CHF). The criterion for inclusion in this study was acute dyspnea on admission from the emergency department (ED), and the exclusion criterion was chest pain. We collected information about each patient's final diagnosis and outcome from the medical file. RESULTS: Overall, the study included 236 patients (100 admitted into the pneumology department and 136 into cardiology). Their mean age was 77.3+/-12.4 years and the sex ratio 1.29 (133 M/103 F). CHR was the final diagnosis for 170 patients (72%: 56% in pneumology and 84% in cardiology). Initial misdiagnosis led to erroneous orientation of 66 patients, 13 (5.5%) of whom were among the 39 (17%) patients who died. The diagnostic accuracy of the specialists' clinical judgment was 74%, compared with 87% and 91% for BNP cut-off levels of 100 and 250 pg/mL, respectively. COMMENT: Without BNP testing in the ED, more than a quarter of the patients with acute dyspnea were incorrectly oriented. Our study found BNP testing to be more effective than clinical judgment. In this series of patients, the cut-off value providing the best sensitivity and specificity was 250 pg/mL. PMID- 17086117 TI - [Troponin I values during rhabdomyolysis in elderly patients admitted to the emergency department]. AB - OBJECTIVE: The objective of this study was to assess the utility of determining serum cardiac troponin I (TcI) levels in the elderly. METHODS: During the nine month study period, all patients older than 70 years admitted to the emergency department presenting rhabdomyolysis (defined by creatine kinase>500 IU/L) were included in this prospective descriptive study, except for those with acute coronary syndrome or pulmonary embolism. Patients were classified into two groups according to their serum TcI level:>0.15 or0.15 ng/mL group and 37 in the other group. Clinical and laboratory indicators were similar in the two groups. In contrast, significantly more patients in the TcI>0.15 ng/mL group had been treated with a curative dose of heparin (14 versus 2, p=0.01). This difference between the two groups was noted in both the emergency department and other hospital units. Serum TcI levels were not correlated with creatine kinase levels. Concordance between emergency department diagnosis and discharge diagnosis was 95%. No patient was discharged with a diagnosis of acute coronary syndrome; one patient, with a serum TcI level0. 15 ng/mL group versus 8% in the other group). Mortality at six months was higher among patients with an elevated serum TcI level, but the difference was not significant (23% versus 8%, p=0.07). CONCLUSION: This study indicated a trend toward higher mortality among patients with elevated serum TcI levels, although the factors underlying these increased concentrations remain unclear. PMID- 17086118 TI - [Oral hygiene for the institutionalized elderly]. AB - INTRODUCTION: The poor oral health of the institutionalized elderly is often underestimated, despite the seriousness of the complications it causes. OBJECTIVES: This descriptive cross-sectional study aims to evaluate the oral health needs of a representative sample of residents of institutions for the elderly in the Limousin and Poitou-Charentes regions. METHOD: This study assessed clinical indicators of need for dental prevention or treatment for 756 residents in 152 establishments (563 women and 193 men, mean age 83+/-9 years), noting the level of oral hygiene and access to treatment at each center. Directors and staff were also asked for suggestions to improve the situation. RESULTS: At least one indicator of need for prevention or treatment was found for 86% of residents, and 91% of this group was not currently receiving treatment. Staff suggestions confirmed the high level of need reported. DISCUSSION: Caregivers in institutions for the elderly must receive training in the provision of oral hygiene measures, oral examinations should be performed routinely on admission to each establishment, and treatment networks should be developed to meet the oral health needs of the institutionalized elderly. PMID- 17086119 TI - [Esophageal leiomyoma. Six cases of video-assisted enucleation]. AB - AIM OF THE STUDY: To report our experience with video-assisted enucleation of esophageal leiomyomas. METHOD: Retrospective analysis of six (five men and a woman) patients who underwent video-assisted submucosal tumor enucleation (4 with thoracoscopy and 2 with laparoscopy). RESULTS: Only one patient had postoperative complications: a parietal hematoma at a trocar site and a fever with right pleural, resolved after pleural drainage. Outcome was satisfactory for all patients, and no relapses have been noted. CONCLUSION: Video-assisted surgery offers incontrovertible advantages for the treatment of this benign pathology. PMID- 17086120 TI - [Early maternal-fetal transmission of the Chikungunya virus]. AB - INTRODUCTION: Since the onset of the Chikungunya outbreak in Reunion Island, vertical maternal-fetal transmission of the virus has been observed in newborns, but no such transmission has been demonstrated early during pregnancy. We report here the first three cases of maternal-fetal transmission of the Chikungunya virus (CHIKV) before 16 weeks' gestational age. CASES: Maternal infections occurred at terms of 12 weeks and 4 days, 15 weeks and 5 days, and 15 weeks and were confirmed by positive findings for specific anti-CHIKV IgM. Fetal deaths were subsequently observed, and at that point, CHIKV RT-PCR was negative for all three maternal blood samples. Amniocentesis preceded rupture of membranes in all three cases. RT-PCR showed viral genome in the amniotic fluid of the three fetuses, in the placentas of two, and in the brains of two. Autopsy found no malformations, and all other bacterial and viral test results were negative. DISCUSSION: These findings demonstrate early maternal-fetal transmission of CHIKV, which is suspected to be directly linked to the fetal deaths. This vertical transmission, probably abortifacient, should be considered in the light of human and animal responses to other arboviruses. PMID- 17086121 TI - [Ureteral stenosis in Wegener's granulomatosis. Case report]. AB - INTRODUCTION: Ureteral stenoses in Wegener's granulomatosis are rare. They usually involve the pelvic portion of the ureter and are caused by vasculitic lesions or granulomatous inflammation. CASE: A 38-year-old woman with Wegener's granulomatosis was treated with corticosteroids and monthly intravenous cyclophosphamide pulses. After 4 months, urinary retention developed, accompanied by lumbar pain, associated with protenuria and hematuria, and related to bilateral ureteral stenoses. Treatment by endoscopic dilatation and double J stents led to with clinical and radiological improvement, while the medical treatment continued. CONCLUSION: Hematuria in patients with Wegener's granulomatosis suggests renal involvement, but ureteral stenoses must also be considered when these patients present hematuria or urinary tract infections. Surgery should be reserved for those patients in whom medical treatment is not rapidly effective. PMID- 17086122 TI - [Sinonasal mucosal malignant melanoma]. AB - INTRODUCTION: Mucosal melanomas are rare tumors, and primary sinonasal malignant melanomas uncommon tumors that are frequently misclassified. Differential diagnosis should rule out other sinonasal malignancies, including carcinoma, lymphoma, sarcoma, and olfactory neuroblastoma. CASE: We report two new cases of sinonasal melanoma. One involved a 68-year-old man being followed for chronic sinusitis, despite antibiotics; he presented a tumor of the right nasal cavity. The second case was that of a 77-year-old woman, referred for painful gingival swelling shown to involve osteolytic growth in the left maxillary sinus. In both cases, pathologic examination of the biopsy samples concluded that these were sinonasal malignant melanomas. DISCUSSION: Histologically, the diagnosis is very difficult because of the polymorphic features of this tumor, and immunohistochemical studies are very important. The relative inaccessibility of the mucosa to self-examination often delays diagnosis and leads to late detection and poor prognosis. PMID- 17086123 TI - [Sarcoidosis in tattoos in chronic hepatitis virus C]. PMID- 17086124 TI - [Fibromyalgia: rheumatologic or psychiatric disease?]. PMID- 17086125 TI - [Patient and disease: two separate entities]. PMID- 17086126 TI - [Fibromyalgia: a disease of psychic trauma?]. AB - Chronic unexplained pain may be a somatic manifestation of psychological distress - often untreated distress. The association between psychic trauma, posttraumatic symptoms, psychic dissociation, and somatoform disorders is currently well documented. When examining a patient with chronic pain syndrome, it is important to consider its psychic dimension early on and to look for a history of psychic trauma. This can help avoid prolonged chronic effects and the emergence of psychiatric comorbidity. There is currently no consensual medication strategy for treatment of unexplained chronic pain syndrome. Multidisciplinary outpatient management is necessary in these complex cases, which require simultaneous medical and psychiatric referrals. PMID- 17086127 TI - [Air pollution and cardiovascular and cerebrovascular disease. Pathophysiologic basis and groups at risk]. AB - The pathophysiologic basis of the harmful action of air pollutants rests principally on their proinflammatory and prothrombotic effects. The principal pollutants currently identified as responsible for these effects are fine suspended particles (PM2.5) and ozone. Inflammatory effects, initially localized in the lungs, then become systemic. Analysis of groups at the highest risk of atherothrombotic events suggests the elderly may be most sensitive to the effects of air pollution. PMID- 17086128 TI - [Interactions between cardiovascular drugs and anesthesia and surgery]. AB - Decisions about chronic treatments during the perioperative period must be made at the presurgical anesthesia consultation. It is increasingly rare to stop treatment during this period, because: This new rule is applied particularly to patients with cardiovascular disorders. Beta blockers have a special role in preventing the onset of postoperative cardiovascular events. The role of statins requires further precision but they appear to fit into the same preventive approach. Interruption of antiplatelet agents appears to be associated with a risk of arterial thrombosis in patients with coronary conditions, notably those with conventional stents and most especially those with drug-eluting stents. PMID- 17086129 TI - [Devic disease]. AB - Devic disease, also known as neuromyelitis optica, is a severe rare condition characterized clinically by one or more episodes of optical neuritis and myelitis. Pathologically, it is characterized by extensive demyelination associated with axon loss and deposits of complement and immunoglobulins (IgM) within the lesions. Specific antibodies for this disease (IgG NMO) were recently identified. Immunosuppressive treatment is currently the best option for preventing relapse. PMID- 17086130 TI - [Regional conciliation and compensation commissions: how they work]. AB - The Law of 4 March, 2002, established regional commissions of conciliation and indemnification (CRCI) to enable the swift and amicable resolution of demands for damages due to medical care (including prevention and diagnosis) by private practitioners, private clinics, and public hospitals. The commission may decide that: (1) the physician or healthcare facility is liable (generally, indemnification by the insurance company), or (2) that indemnification is due in the name of 'national solidarity' (indemnification by a national organism called ONIAM), or (3) no damages are due. There are two important restrictions: For the CRCI to have jurisdiction to determine indemnification, it is imperative that at least one of the severity criteria be met. The 4 severity criteria are: (1) a permanent disability greater than 24%, (2) temporary disability for at least 6 months during a one-year period, or (3) permanent inability to perform the previous occupation, or (4) rarely, especially severe problems (for example, economic) in living conditions that result from the medical care. The procedure applies only to care provided since 5 September 2001. From the request for indemnification by 'ONIAM or the insurer, the process must last no longer than one year. PMID- 17086131 TI - [The tuberculin test today]. PMID- 17086132 TI - [BCG vaccination: typically French quarrel or real debate?]. PMID- 17086133 TI - [Epidemiology of tuberculosis in France]. AB - Tuberculosis causes an ever-increasing public health burden throughout the world and is one of the World Health Organization's top priorities. In France, the incidence of tuberculosis has been stable at around 10 cases per 100 000 inhabitants per year since 1997. In 2004, the incidence rate in 2004 was 9.2 per 100 000, with substantial geographic disparities: the Ile-de-France region (including Paris and its suburbs) accounted for more than 40% of all reported cases (20.8 per 100 000). Incidence rates of tuberculosis vary according to the geographic origin: in France, this rate is highest among people from sub-Saharan Africa (incidence rate: 166.7 per 100.000 compared with 5.0 per 100.000 for those born in France). Pulmonary tuberculosis is the disease form most commonly observed (71.5% of cases). Among those cases, 76.9% were laboratory-confirmed in 2004. National guidelines to improve case management and disease control were updated in 2003 and 2005. Funding must be made available to reinforce tuberculosis surveillance and to conduct public health initiatives targeted at the populations most at risk. PMID- 17086134 TI - [BCG Today]. AB - Until recently, French BCG vaccination policy required BCG vaccination before entry into day-care centers or school and the revaccination of tuberculin negative children. Recent reassessment of this policy led in 2004 to the discontinuation of all revaccinations and of postvaccination tuberculin tests except those performed as part of tuberculosis diagnosis or in the monitoring of healthcare and social workers, for whom BCG vaccination remains mandatory. After an epidemiologic assessment of the effect of either selective vaccination of high risk children or discontinuation of all BCG vaccination and analysis of balance of risks and benefits of each option, the High Council of Public Hygiene (Infectious diseases section) approved the principle of selective vaccination of children. As a prerequisite to this implementation of this strategy, however, the council advised the Ministry of Health that other control measures to decrease the risk of tuberculosis in children should be reinforced. This issue is made more complex by the withdrawal in 2006 of the multipuncture vaccine, used in France in more than 90% of primary BCG vaccinations. PMID- 17086135 TI - [Modern diagnosis of tuberculosis]. AB - Microscopic examination of sputum smears (search for acid-fast bacilli or AFB) is the most rapid procedure for diagnosis of contagious tuberculosis. Gene amplification is not yet reliable for direct detection of M. tuberculosis in clinical specimens that are AFB smear-negative. Culture (3 to 8 weeks on Lowenstein-Jensen medium or 1 to 4 weeks in liquid media) remains essential to identify AFB and conduct antibiotic susceptibility testing. AFB from culture can be identified in a few hours by molecular approaches with specific DNA probes. Results of susceptibility testing, even in liquid media, are not available until 2 to 4 weeks after the recovery of specimens, although mutations of the rpoB and katG 315 genes, which confer resistance to rifampin and isoniazid, can be detected within hours by molecular hybridization with specific probes fixed on strips. Immunologic tests that measure the interferon gamma produced by sensitized lymphocytes are promising tools for the diagnosis of latent tuberculosis. PMID- 17086137 TI - [Management of tuberculosis in France: guidelines of the French Pulmonary Society]. PMID- 17086136 TI - [Paradoxical reactions in tuberculosis]. AB - Paradoxical reactions during tuberculosis are defined by exacerbation of clinical symptoms or laboratory or radiologic abnormalities during antituberculosis treatment or when an antiretroviral treatment is added to it. This phenomenon does not involve relapse of the opportunistic infection but rather a reconstitution of an abnormal immune response. The phenomenon is relatively frequent after antiretroviral treatment begins in HIV-infected patients with tuberculosis. Diagnosis, based on a group of heterogeneous clinical and/or radiologic criteria, is often difficult. Although outcome is most often spontaneously favorable, complications are possible. Treatment relies on the maintenance of specific treatment of the pathogen responsible for the opportunistic infection, the antiretroviral treatment, and possibly the addition of corticosteroid therapy. PMID- 17086138 TI - [Trends in tuberculosis treatment duration]. AB - The two principal characteristics of tuberculosis treatment are its length (several months) and the need to use several antibiotics simultaneously (multiple drug therapy). Multiple drug therapy is intended to prevent the selection of resistant mutants at the beginning of treatment, when the bacilli population is largest. The length of treatment is due to dormant bacilli, which are much more difficult for antibiotics to kill than actively multiplying bacilli are. Rifampin and pyrazinamide are the most potent drugs against these dormant bacilli. The so called sterilizing activity of rifampin has reduced the duration of treatment from 18 to 9 months, and the contribution of pyrazinamide reduced this time still further, to 6 months. When one of these drugs cannot be used because of resistance or toxicity, duration of treatment increases to the earlier levels. In the extreme case of multidrug-resistant tuberculosis where neither isoniazid nor rifampin can be used, and sometimes even not pyrazinamide, treatment is recommended for 18 to 24 months. New antituberculosis drugs under development allow us to envision further reduction in the duration of treatment of both drug resistant and drug-sensitive tuberculosis. PMID- 17086139 TI - [Tuberculosis during treatment by TNFalpha-inhibitors]. AB - The clinical forms of tuberculosis that occur during anti-TNFalpha treatment are frequently extrapulmonary or even disseminated and life-threatening. The paradoxical reactions that can occur under appropriate treatment after stopping TNFalpha inhibitors raise the question of an immune restoration phenomenon. Adverse drug reaction reporting and epidemiologic studies, despite their methodological limitations, appear to show an excess risk of tuberculosis. Experimental studies reinforce these data. The French drug agency (Afssaps) has issued guidelines for the prevention and management of tuberculosis occurring under anti-TNFalpha treatment. Analogous guidelines in Spain led to a reduction in the incidence of these cases. PMID- 17086140 TI - [Part I: Face recognition]. AB - Faces represent a crucial vector of interhuman communication. The message transmitted by the face has multiple features. Recognition of each feature can be impaired independently or in combination with others. In order to understand the behavioral consequences of such impairments, which can be a major social handicap, we first must specify the neural networks involved in face recognition. We propose in this first part to present the systems involved in face recognition, in particular the question of identity and prosopagnosia. Different neural networks are indeed implicated in the recognition of invariant facial features such as identity, gender, ethnicity, and recognition of variant features like facial expression and eye gaze. This paper is illustrated by some of our scalp and intracranial electrophysiological studies performed in humans allowing us to describe some aspects of face recognition dynamics combining an excellent spatial and temporal resolution. Intracranial recordings were performed in drug refractory epileptical patients implanted with depth electrodes. These studies demonstrate that numerous deep brain and cortical structures participate early and sometimes in a sustained manner in face recognition. PMID- 17086141 TI - [Part II: Recognising facial expressions]. AB - In this second part, we address particularly the question of the neural mechanisms and structures involved in the recognition of facial emotional expressions that are crucial in social cognition. Emotion recognition in others can be critically impaired in some neurodegenerative and neurovascular diseases. That dysfunction sometimes correlated to disabling behavioural disorders and interpersonal communication impairment must be further understood. The results of a series of scalp and intracranial event related potential recordings, as well as recent advances in the literature, are reported. ERPs to facial emotional expressions were thus recorded in multiple subcortical and cortical areas in drug refractory epileptical patients implanted with depth electrodes. The roles of amygdala, insula and prefrontal cortex located at crossroads between perceptive analysis and emotional conceptual knowledge are particularly underlined. Altogether, these studies demonstrate that facial expressions are widely processed in space and time, some structures reacting very early and automatically, others providing a sustained reaction depending on the attention. PMID- 17086142 TI - [Sporadic cerebral amyloidotic angiopathy]. AB - INTRODUCTION: Sporadic cerebral amyloid angiopathy (CAA) is a microangiopathy identified by neuropathological examination in more than 30 percent of patients over 85 years of age. STATE OF ART: Boston criteria for diagnosis of CAA--related hemorrhage are as follows: "definite CAA", "Probable CAA with supporting pathology", "Probable CAA" and "Possible CAA". Clinical manifestations of CAA are either lobar, cortical, corticosubcortical or cerebellar hemorrhages associated with progressive dementia. Dementia, corresponding either to Alzheimer disease, vascular or mixed dementia, precedes hemorrhages in 25 to 40 percent of cases. Brain MRI can demonstrate microbleeding. PERSPECTIVES: This review compares data regarding CAA prevalence, intracranial hemorrhages, and their risk factors in old patients. Diagnosis and preventive strategies are discussed. It would be useful to identify those affected by CAA among elderly demented patients with atrial fibrillation requiring anticoagulation therapy. CONCLUSIONS: CAA is suspected in the presence of recurrent lobar or cerebellar hemorrhages, and moreover if associated with pre-existing dementia. In elderly demented patients, MRI criteria to detect CAA should be considered in order to prevent hemorrhage risk, particularly after anticoagulation therapy. PMID- 17086143 TI - [Long-term outcome in patients with symptomatic low-grade oligodendrogliomatous tumors treated by cytotoxic agents]. AB - INTRODUCTION: Whether aggressive treatment or no treatment is the optimal management for low-grade gliomas is controversial. However, symptomatic low-grade gliomas require prompt therapeutic intervention because of neurological impairment, uncontrolled seizures, and deterioration of life quality. METHODS: We report the long-term follow-up, 71 months, of seven patients treated by procarbazine, lomustine and vincristine (PCV) therapy for a symptomatic low-grade oligodendrogliomatous tumor. The mean age at diagnosis was 47 years, the mean time from first symptoms to initiation of PCV therapy was 62 months (range 15 147). RESULTS: All patients initially responded favorably, with improvement of the neurological symptoms and radiological response. Chemotherapy was clinically well tolerated, the main side effect being low hematological toxicity. During the follow-up, no progression was observed in two patients. For the five remaining patients, the time to progression after the PCV induction was 56+/-12 months (range 38 to 73). Four of these patients showed favorable response to a second line of treatment. CONCLUSION: PCV therapy is an interesting therapeutic option for progressively symptomatic low-grade gliomas, even in cases with large tumoral volume. This treatment, of moderate toxicity, improves the quality of life and can result in long-term tumor stabilization. PMID- 17086144 TI - [Neurological manifestations of type 1 Gaucher's disease: Is a revision of disease classification needed?]. AB - INTRODUCTION: Gaucher's disease (GD), the most prevalent inherited lysosomal storage disorder, is caused by deficient glucocerebrosidase activity. The resulting accumulation of glucocerebrosides in lysosomes of macrophages leads to hepatosplenomegaly, anemia, thrombocytopenia, and various bone manifestations. Gaucher's disease is classified into 3 types based on the nature of its effects on the central nervous system. Type 1, the most common variant, is classically nonneuronopathic. However, the occurrence of Parkinsonism seems to be more frequent in type I Gaucher's disease than in the general population. Furthermore, heterozygotes for certain glucocerebrosidase gene mutations have a higher risk to develop Parkinson's disease. OBSERVATIONS: We report our experience about 9 patients with Gaucher's disease and their association with neurological manifestations. CONCLUSION: These recent data may discuss Gaucher's classification and the existence of a continuum between neurologic and non neurologic forms of the disease. PMID- 17086145 TI - [Neurological involvement in Behcet's disease. 154 cases from a cohort of 925 patients and review of the literature]. AB - INTRODUCTION: Neurological involvement is well described in Behcet's Disease (BD), with variable prevalence of 5.3 to 30p.cent. The purpose of this retrospective study was to analyze the clinical patterns of neuro-Behcet (NB) and to compare them with different clinical features of 925 BD registered in the same period and the literature. METHODS: All patients of NB fulfilled the International Study Group Criteria for the diagnosis of BD. 93 patients with headache, without other neurological symptoms were excluded. The findings were supported by cerebrospinal fluid, computed tomography scan, magnetic resonance imaging and angiography. RESULTS: NB was present in 16.64p.cent of BD. A sex ratio male/female: 4.31. The mean age of patients with NB was 31.76 years. The average age of onset of NB was 29.83. The mean duration of the NB disease was 3.81 years. The findings were categorised in 2 main types: parenchymal and non parenchymal involvement. Some patients had features of both types (mixed patterns). 94 patients (61.03p.cent) presented with parenchymal central nervous system (CNS) involvement. The most common findings were pyramidal signs, cranial nerve palsies, pseudobulbar syndrome and cerebellar signs. 27 patients (17.53p.cent) without parenchymal CNS which were divided into: intracranial hypertension in 24 patients (15.58p.cent) presenting headache, vomiting and bilateral papilloedema; cerebro-arterial involvement in 3 cases (one of them had cerebrovascular aneurysms). Mixed patterns were observed in 9 cases. Erythema nodosum and vascular involvement were more frequent in intracranial hypertension than in BD. Other clinical features were reported: pure meningeal pattern in 15 cases, pure peripheral nervous system involvement in 4 cases, isolated cranial nerves in 8 cases, chorea in one patient and pseudotumor of cervical medulla in other patient. 6 cases of juvenile BD and one case of familial Behcet were observed. 133 patients were treated (86.36p.cent). Mean duration of treatment was 7.46 month. We used corticosteroids, immunosuppressive agents (cyclophosphamid, azathioprine, chlorambucil) and anticoagulant in intracranial hypertension. The course of disease was good in only 54.13p.cent of cases, and was bad in 18.79p.cent. CONCLUSION: NB occurred frequently in men and is more serious especially in parenchymal CNS involvement: 7 deaths (6 from parenchymal CNS). PMID- 17086146 TI - [Spinal dural arteriovenous fistula with peri-medullary venous drainage: analysis of a series from a single centre and review of the literature]. AB - INTRODUCTION: The optimal diagnosis and therapeutic managements of spinal dural arteriovenous fistulae, a rare disease, is discussed. MATERIAL AND METHODS: We report a series of 10 patients seen in a 7-year period, treated by embolization. RESULTS: There were 6 males and 4 females, with an average age of 58.4 years (range: 31 to 74 years). Diagnosis was made 2 days to 5 years (mean 15.3 months) after symptom onset with a high rate of incomplete or atypical clinical patterns (prolonged isolated sensory disorder, lumbo-sciatalgia, claudication, monoplegia), or sudden-onset deficit. In all cases, spinal MRI showed an intramedullary high-intensity signal on T2-weighted images but enlarged intradural vessels in only 70p.cent of cases. Angiography showed in all cases an arteriovenous fistula at the upper dorsal level (T1 to T7, 5 cases), lower dorsal (T8-T12, in 3 cases) and lumbar levels (2 cases). The fistula was successfully obliterated after initial embolization in 3 cases, but two (2 cases) or 3 procedures (2 cases) were sometimes required. The endovascular therapy failed in 3 cases. CONCLUSION: In this series, the fistula was successfully obliterated in 70p.cent of patients. In the literature, fistula of 96.8p.cent to 97.9p.cent of patients were obliterated by surgery, which is a more invasive treatment. Embolization could be used as the first-line therapy, but incomplete obliteration requires rapid surgery. While the success of embolization is demonstrated, MRI and angiography must still be performed 6 months later. PMID- 17086147 TI - [Treatment with intravenous rt-PA in a general hospital context: A review of the Saint-Jean Hospital experience from Perpignan, France]. AB - INTRODUCTION: Intravenous recombinant tissue plasminogen activator (rt-PA) has approval for use despite of its authorization for treatment of ischemic stroke within the 3-hour time window in 2003, is rarely used in community hospital (CH). It therefore remains questionable if the positive results of the key studies conducted in specialized centers may be extended to community hospitals less specialized in the management of stroke. METHODS: We report the results of an observational cohort study including 39 patients treated with intravenous rt-Pa (according to the NINDS rt-PA stroke trail treatment protocol) at St Jean Hospital (Perpignan, France) between March 1, 2002 and August 31, 2005. Results are compared to those of the treated arm of the NINDS study. RESULTS: 1.2p.cent of ischemic stroke were treated with intravenous rt-Pa. Results are similar to those of the NINDS study: The outcome was favorable (modified Rankin score (mRS) with 0 or 1) for 44p.cent of the patients (as compared to 39p.cent in the NINDS study (X2 = 0.34; p = 0.5)) and there was no significant difference in term of death or outcome as assessed by mRS at 3 months (X2 = 0.09; p = 0.75 and X2 = 0.77; p = 0.75, respectively). No symptomatic hemmorrhagic transformation related to the use of rt-Pa was observed. CONCLUSION: Our results indicate that rt-PA therapy for ischemic stroke may be as safe and effective in the setting of a community hospital as it is in specialized centers. PMID- 17086148 TI - [Lithium-induced encephalopathy mimicking Creutzfeldt-Jakob disease]. AB - INTRODUCTION: Creutzfeldt-Jakob disease (CJD) has a poor prognosis. Certain clinical presentations can be suggestive yet mimic a curable disease. OBSERVATION: In the present study, we report the case of a 67-year-old man with a one-month history of progressive dementia, with myoclonic jerks and cerebellar syndrome suggesting a diagnosis of Creutzfeldt-Jakob encephalopathy. He had been treated for 9 years with lithium for a bipolar disorder. The results of the different investigations and the favorable course after discontinuation of lithium were in favor of the diagnosis of drug-induced Creutzfeldt-Jakob syndrome. CONCLUSION: This case illustrates the importance for researching a curable etiology in presence of clinical features suggesting a CJD. PMID- 17086149 TI - [Reintroduction of treatment with lamotrigine in combination with valproate after an initial allergic skin reaction]. AB - INTRODUCTION: Lamotrigine (LTG) is effective for a variety of seizure types especially against absence seizures in cotherapy with valproate (VPA). Skin rash is the most frequent side effect leading to LTG discontinuation. OBSERVATIONS: We report two patients with refractory absence epilepsy and a previous history of allergic reaction to LTG in which LTG was rechallenged. LTG was effective in both cases without incident. DISCUSSION: High initial dose and rapid dose escalation is associated with increased rash rate. LTG is contraindicated in patients who have demonstrated hypersensitivity to the drug. However, because the initial dose and the dose titration play a crucial role, LTG was reintroduced with a very low initial dose and very slow titration schedule without recurrence of skin rash. CONCLUSION: Patients who have experienced a previous mild skin allergy may be reconsidered for a rechallenge with LTG if the expected therapeutic response to the drug is high. Patients must be warned about the risks. PMID- 17086150 TI - [Exacerbation of tonicoclonic seizures in a juvenile myoclonic epileptic taking lamotrigine]. AB - INTRODUCTION: Lamotrigine is an anti-epileptic drug considered as a first-line therapy in idiopathic generalised epilepsy. Lamotrigine is well-tolerated and secondary aggravation of epileptic syndromes has rarely been reported. CASE REPORT: We report the case of a 19 year-old man with juvenile myoclonic epilepsy in whom lamotrigine lead to the exacerbation of generalised tonico-clonic seizures, reversible when lamotrigine was stopped and substituted by valproic acid. CONCLUSION: Lamotrogine may induce exacerbation of generalised tonico clonic seizures and myoclonic jerks in juvenile myoclonic epilepsy. PMID- 17086151 TI - [Amyloid cardiomyopathy: A rare cause of cerebral embolism]. AB - INTRODUCTION: We report a case of amyloid cardiopathy revealed by a cerebral embolism. CASE REPORT: A 55-year-old patient was admitted with a right hemiplegia and aphasia due to an infarction in the middle cerebral artery territory. Echocardiography was suggestive of an amyloid cardiopathy, and an IgG lambda multiple myeloma with renal insufficiency was discovered. The patient died suddenly 4 months later after chemotherapy was initiated. CONCLUSION: Embolic complications are rare and late in cardiac amyloidosis. The diagnosis may be suspected by echocardiography. PMID- 17086152 TI - [Reccurent stroke revealing catastrophic antiphospholipid syndrome with hepatitis C viral infection]. AB - INTRODUCTION: Very little cases of antiphospholipid syndrome (APLS) have been described among patients having chronic hepatitis C virus infection (HCV). CASE REPORT: We report the observation of a 43-year-old woman who presented APLS diagnosed following recurrent strokes. Etiological investigations concluded on an association of APLS with HCV infection. DISCUSSION: Besides being exceptional, this association raises certain etiopathogenic problems. Indeed while antiphospholipid antibodies (aPL) are frequently noted during chronic hepatitis C, they rarely generate thromboembolic complications. PMID- 17086153 TI - [The diagnosis and management of familial amyloid polyneuropathy]. AB - INTRODUCTION: Familial amyloid polyneuropathy designates a group of dominantly inherited neuropathies, with extracellular deposition of amyloid substance in various tissues. BACKGROUND: The 3 main precursor proteins encountered in these disorders are transthyretin, apolipoprotein A1 or gelsolin. Among them, transthyretin neuropathies are by far the most frequent type with a severe sensori-motor and autonomic neuropathy as the hallmark of the disease, most often associated with cardiac manifestations. First described in Portugal, the affection was subsequently reported across the world, although Portugal, Japan and Sweden are the 3 main areas of prevalence. In the past years, an increasing number of mutations have been identified in the TTR gene, along with a larger clinical spectrum than initially thought. Variable age of onset and penetrance are also largely reported with unclear phenotypic-genotypic correlations. Indeed, the contribution of the molecular genetics is important to ensure the diagnosis at an early stage, but also for predictive diagnosis, in the setting of genetic counselling. PERSPECTIVES: Over the last 15 years, liver transplantation (LT) has enabled improved prognosis of this devastating condition. FUTURE PROSPECTS: at present, such procedure should be performed in Val30Met patients, as early as possible in the course of the disease. Experience with such procedure in patients with other TTR variants remains scarce. Other therapeutic strategies are awaited. CONCLUSION: This review summarizes the recent data on the diagnosis and management of patients and families affected with TTR amyloid neuropathy. PMID- 17086154 TI - [Medical management of patients with atypical degenerative parkinsonian syndromes]. AB - Atypical degenerative parkinsonian syndromes (progressive supranuclar palsy, multiple system atrophy, corticobasal degeneration, Lewy body dementia) are an important differential diagnosis to idiopathic Parkinson's disease. However, because these disorders are characterized by the degeneration of multiple neuronal populations, treatment approaches are much less specific than in Parkinson's disease, where dopamimetic drugs represent the mainstay of therapy. Thus, and because the progression of these disorders is usually more aggressive than Parkinson's disease, many physicians face a form of therapeutic resignation when confronted with patients suffering from atypical degenerative parkinsonian syndromes. However, in the present article, we wish to show that a symptom-by symptom approach can substantially relieve the patients and their caregivers by providing an overview of pharmacologic and non-pharmacologic treatment options. PMID- 17086155 TI - Catalases of Aspergillus fumigatus and inflammation in aspergillosis. AB - The article describes various features of aspergillosis and a discussed the role of calatases produced by Aspergillus fumigatus during infection. Since a large body of invasive Aspergillus infection occurs as an opportunistic infection in variously impaired defense mechanisms, there is a wide spectrum of histopathological features of lesions demonstrated at the site of infection. Accordingly, histopathology of the lesions can be understood as a phenotypical representation of interaction between differently impaired functions of neutrophils and macrophages and virulence factors of invading Aspergilli. Consideration of previous pathological knowledge regarding infection and inflammation provides much important information to predict the pathophysiology of a patient. Meanwhile, detoxification of hydrogen peroxide by catalases has been proposed as a way to overcome this host response. A. fumigatus produces three active catalases, one from conidia and two from mycelia. CatAp, a spore specific monofunctional catalase, is resistant to heat and metal ions. In spite of their increased sensitivity to H(2)O(2), killing of catA conidia by alveolar macrophages, virulence in animals was similar to wild type conidia. In contrast to mycelial Cat1p, and CatAp catalases, the mycelial Cat2p is a bifunctional catalase-peroxidase enzyme and is also sensitive to heat, metal ions and detergent. Surprisingly, the mycelium of the double cat1 cat2 mutant with no catalase activity has only a slightly increased sensitivity to H(2)O(2) and was as sensitive to the killing of polymorphonuclear neutrophils as the wild type strain. However, it showed a delayed infection in the rat model of aspergillosis compared to the wild type strain. Consequently, it should be emphasized that conidial catalase is not a virulence factor but that mycelial catalases transiently protect the fungus from the host defence reactions. PMID- 17086156 TI - [Cell cycle control and CDC28/Cdc2 homologue and related gene cloning of Cryptococcus neoformans]. AB - In Cryptococcus neoformans the DNA content of cells having tiny buds varied rather widely, depending on growth phases and strains used. Typically, buds of C. neoformans emerged soon after initiation of DNA synthesis in the early exponential phase. However, bud emergence was delayed to G2 during transition to the stationary phase, and in the early stationary phase budding scarcely occurred, although roughly half of the cells completed DNA synthesis. The timing of budding in C. neoformans was shifted to later cell cycle points with progression of the growth phase of the culture. Similarly, a deficit in oxygen was demonstrated to delay the timing of budding, prolong the G2 phase and cause accumulation of cells after DNA synthesis, but before commitment to budding. The C. neoformans homologue of the main cell cycle control gene CDC28/Cdc2 was isolated using degenerate RT-PCR. The full-length coding region was then amplified using primers to target the regions around the start and stop codons. The gene was called CnCdk1 and was found to have high homologies to S. cerevisiae CDC28 and S. pombe cdc2. To determine its function, its ability to rescue S. cerevisiae cdc28-temperature sensitive mutants was tested. S. cerevisiae cdc28-4 and cdc28-1N strains transformed with the pYES2-CnCdk1 construct exhibited growth at the restrictive temperature. Results of the sequence analysis and the ability of CnCdk1 to complement the S. cerevisiae cdc28-ts mutations support its assumed role as the CDC28/cdc2 homologue in C. neoformans. PMID- 17086157 TI - [Quorum-sensing system in Candida albicans]. AB - The bacterial behavior system controlled by the cell density is described as quorum-sensing. The system is triggered via autoinducers. Various kinds of autoinducers have been identified from different bacteria. Quorum-sensing signals via autoinducers are involved in regulation of important virulences such as exotoxin, protease, and pigment production. Therefore, this system in pathogenic bacteria has a critical role in the regulation of bacterial pathogenicity. In the pathogenic fungus Candida albicans, an extracellular quorum-sensing molecule that regulates hyphal formation by this organism has been identified in recent years. Candida albicans has been shown to form biofilm on many medical devices, therefore quorum-sensing in this organism has been especially focused on from the aspect of biofilm formation. PMID- 17086158 TI - [Essential genes as potential targets of antifungal agents in pathogenic yeast Candida]. AB - An important point in the development of an antimicrobial agent is whether its target molecules are essential for growth of the microorganism. From this viewpoint, we focused attention on essential genes as potential targets of antifungal agents in the pathogenic yeast Candida. Here we introduce recent attempts for screening, identification, and characterization of essential genes from a haploid yeast Candida glabrata, using temperature-sensitive mutants. Our experimental results suggesting the essentiality of C. albicans PHO85, the homologue of which is known as a negative regulator of the PHO system and as a non-essential gene in Saccharomyces cerevisiae are also described. PMID- 17086159 TI - Amino acid residues affecting drug pump function in Candida albicans--C. albicans drug pump function. AB - Membrane-located drug transporters are important components in the multidrug resistance of microbial cells and human tissues. In fungi, clinically important resistance to antifungal drugs most often results from the over-expression of efflux pump proteins in the plasma membrane of the resistant cell. This review describes studies of the ATP binding cassette (ABC) family of membrane efflux pumps in the opportunistic human pathogen Candida albicans and, in particular, examines how changes in the polypeptide sequence can affect pump function. The identification of amino acid residues affecting pump function can provide new insights into efflux pump mechanisms and the relationship between structure and function. Such information will be important for the design of pump inhibitors which could supplement existing antifungal drugs. PMID- 17086160 TI - [Use of PCR based diagnosis for common invasive fungal infections in the intensive care unit]. AB - Deep-seated Candida infections and invasive aspergilloma are becoming a serious problem for individuals who need intensive care. The laboratory diagnosis of such infections is sometimes delayed due to relatively slow growth of these yeasts from clinical specimens. Several studies seem to indicate that early detection of deep-seated and invasive fungal infections is possible using genomic amplification methods. In the present study, we used a novel PCR assay that can assay five clinically common species (C. albicans, C. parapsilosis, C. tropicalis, C. glablata, and A. fumigatus) simultaneously. We evaluated the utility of this PCR based diagnosis with seven patients with candidiases. This assay is more sensitive than the culture result in 26 clinical samples (chi(2)=5.16, p < 0.05). In the clinical course of each patient, the number of detected fungal species gradually increased. More than two species were detected from single or several clinical specimens, and these patients would die within 14 days compared with the 61 day period individuals with zero or one species would live (p < 0.005). Before super infections of fungus, an antifungal drug could be applied to a suspected patient in the ICU. To improve sensitivity of this diagnosis from blood samples, we evaluated them after one day incubation at 34 degree. We found a PCR product in 10 of 20 blood samples taken from five children after bone marrow transplantation. One of four negative samples became positive after more than 48 hours of incubation. PMID- 17086161 TI - [Invasive fungal infection in solid organ transplantation: toward evidence-based prophylaxis and preemptive treatment]. AB - Although the opportunity to discuss infectious complications in solid organ transplantation is increasing in Japan as elsewhere, the length of clinical experience in extra-renal transplantation is still short and even experience in living donor organ transplant is very limited except for those involving the kidney or liver transplantation. Risk of invasive fungal infection in organ transplant recipients is highly dependent on the immunocompromised status accompanying end-stage organ failure before transplant operation and on the resultant history of infectious complications. These factors as well as surgical and postoperative should be incorporated in a systematic and dynamic manner to evaluate risk of invasive fungal infection. In addition to prophylactic management based on such risk evaluation, it is desirable that preemptive treatment be started on quantification of clinical symptoms, imaging diagnosis, screening culture, and serological indices. Emergence of newer and more potent antifungal agents with lower toxicity potentially changes the concept of antifungal treatment. On the other hand, early and impression-oriented preemptive treatment has tended to increase. It is still questionable whether the knowledge obtained from Western experience can be directly applied to solid organ transplant medicine in Japan. Extensive and detailed clinical experience is mandatory to pursue diagnosis, epidemiology, and risk factors in Japan and establish our criteria for prophylactic and preemptive use of antifungal agents. PMID- 17086162 TI - [Efficacy of micafungin sodium on fungal infection in critical care]. AB - The clinical effects and tolerability of micafungin sodium in daily practice for the treatment of fungal infection in critically ill patients were evaluated in an open-labeled, non-comparative, observational study. All patients admitted to intensive care units (ICUs) of 3 hospitals in Chiba prefecture between June 2003 and March 2005, who were treated with micafungin because of known or suspected fungal infection, were included in the study. A total of 34 patients received micafungin and 29 cases of them were subjected to analysis. Fungal infections were classified as "proven" in 3 patients (10.3%) and "possible" in 26 (89.7%). Candida was detected in 16 patients, most of them were Candida albicans and 4 cases were non-albicans Candida. Clinical effects of micafungin were "cured" and "improved" in 20 patients (77%), "failure" in 6 (23%), and "undetermined" in 3 cases. Adverse events were reported in 10 patients, but there was no significant event. In conclusion, micafungin was effective in 77% of proven or suspected fungal infections in critically ill patients admitted to the ICU. The drug was well tolerated and discontinuation of its treatment due to adverse events was not experienced during the study period. PMID- 17086163 TI - [In vitro activities of antifungal drugs against clinical isolates of Trichophyton tonsurans]. AB - To determine drug susceptibility of Trichophyton tonsurans endemic in Japan, in vitro MICs of antifungal drugs against a total of 10 clinical isolates of T. tonsurans collected from dermatophytosis patients were measured by the agar dilution method and the broth microdilution method. The agar dilution method was not appropriate as the growth of T. tonsurans on the agar medium was too slow to determine drug activity, while the broth microdilution method was thought to be an appropriate method for this study. The MIC90 values determined by the broth microdilution method for terbinafine, itraconazole, miconazole and ketoconazole were 0.013, 0.1, 0.8 and 0.4 microg/ml, respectively. Meanwhile, the MIC90 values of lanoconazole and luliconazole, known to be antifungal drugs potent against dermatomycosis, were 0.00078 and 0.00039 microg/ ml, respectively. The drug susceptibility of these T. tonsurans isolates to the aforementioned antifungal drugs was found to be on a similar level with that of T. mentagrophytes and T. rubrum, major causative agents of dermatomycosis. The results also demonstrated the strong antifungal activity of lanoconazole and luliconazole against T. tonsurans. PMID- 17086164 TI - Isolation of Fonsecaea pedrosoi from the shell of the babassu coconut (Orbignya phalerata Martius) in the Amazon region of Maranhao Brazil. AB - Fonsecaea pedrosoi, a dematiaceous fungus and the main causative agent of chromoblastomycosis, has been isolated in worldwide from different natural sources in regions where the disease is endemic. In the Amazon region of Maranhao, Brazil, where the disease is prevalent, the breaking of the babassu coconut (Orbignya phalerata Martius) represents an important agricultural activity. In order to determine the presence of this fungus on this plant and on other natural substrates, material was collected in the Fortaleza Village Municipality of Pinheiro, Maranhao, in April and September 2002. A total of 68 samples, including 18 (26.5%) obtained from the shell of the babassu coconut, were analyzed. Samples were cultured using a standard method. Isolates were identified based on macromorphological aspects of the colonies on Sabouraud dextrose agar and based on the micromorphology of the conidia after growth on potato dextrose agar. Exophiala sp. was the most prevalent fungus isolated from the different natural substrates analyzed, while Cladophialophora sp. was only isolated from decomposing wood. Fonsecaea pedrosoi was isolated from one sample of babassu coconut shell suggesting that this coconut represents an important source of infection of chromoblastomycosis during extraction of the plant product in this region. PMID- 17086165 TI - The effect of dimethyl sulfoxide (DMSO) on the growth of dermatophytes. AB - Dimethyl sulfoxide(DMSO) is frequently used as a solvent for antifungal drugs in various studies to determine their MICs. Reports on comparative evaluation of methods for the susceptibility testing of antifungal drugs have shown there is poor agreement among methods. Besides other factors which could cause variability in the results, one important factor might be the effect of DMSO on the growth of fungi. The effect of DMSO on the growth of some species of Candida has been reported in the literature. The present study aimed at determination of the effect of different concentrations of DMSO (0.125 to 10%) on the growth of dermatophytes by agar diffusion method. There was no growth of fungi in 10% DMSO, between 1.25 and 5% there was a rather linear dose-related inhibitory effect on the growth, significantly different from the controls, and below 1% there was a variable effect among the species. DMSO down to 0.25% significantly inhibited the growth of most strains of M. canis. The lower concentrations of DMSO, which apparently do not affect the growth of fungi, may potentiate the effect of antifungal drugs. PMID- 17086166 TI - [Trichophyton tonsurans infection among judo practitioners who attended the National Junior High School Judo Tournament in Japan (2005): incidence and therapeutic response]. AB - The spread of Trichophyton tonsurans infection among high school students and university undergraduates who practice Judo is an emerging problem in Japan and other countries. However, the extent of infection among Judo practitioners in junior high school in Japan is unknown. We conducted an epidemiological study of T. tonsurans infection among students who participated in the national junior high school Judo tournament in 2005. Of the 1,039 tournament participants invited to undergo screening, 496 (218 boys and 278 girls) consented, and 45 participants (9.1%) were found to be positive by hairbrush culture. We found the following to be relative risk factors for T. tonsurans infection: 1) male gender, 2) frequent judo practice in groups at either a high school or a dojo, 3) presence of tinea corporis in practice partners, 4) history of tinea corporis, 5) classification in lower-weight categories. 45 culture positive subjects were offered treatment and re-examined by hairbrush culture 3 months later. All twelve of them had negative cultures after miconazole shampoo treatment. A half of 12 subjects who had systemic antifungal therapy with itraconazole had positive culture. These observations suggest that T. tonsurans infection is rapidly spreading among junior high school Judo players in Japan. We speculate that the outbreak is caused, at least in part, by ignorance of the disease among Judo students, coaches and officials due to the high incidence of carriers and the mild or asymptomatic form of disease seen in infected individuals. Appropriate measures should be taken immediately to prevent more severe outbreak of this disease. PMID- 17086167 TI - Using contrast sensitivity to estimate the cost-effectiveness of verteporfin in patients with predominantly classic age-related macular degeneration. AB - AIMS: To re-evaluate the cost-effectiveness of photodynamic therapy with verteporfin (Visudyne, Novartis AG, Switzerland) in patients with predominantly classic and classic choroidal neovascularization (CNV) owing to age-related macular degeneration (AMD), using new evidence on the impact of contrast sensitivity on health status. METHOD: A health economic model is used to synthesise the evidence on contrast sensitivity and treatment rates from the TAP Investigation with health state utilities and costs. Impairment of visual function is estimated using a Markov model to predict transitions between states of contrast sensitivity. Each state is associated with costs and a health state utility. Total expected costs and benefits for a cohort of patients over a defined number of cycles are calculated. The expected health state utility for each disease state was estimated using results from a study of 209 patients with AMD in Sheffield. The model includes the costs associated with treatment and monitoring in the verteporfin treatment arm and costs offset by delaying the deterioration of visual function. RESULTS: Beyond 3 years, the annual costs of the verteporfin arm are estimated to be less than the annual costs of the control arm, owing to the cost associated with higher blindness prevalence in the control arm. Over time, the results show that both the incremental utility and cost decreases. By 10 years, the estimated incremental cost-effectiveness is approximately pound20 996 per Quality-Adjusted Life Years. CONCLUSION: The results of this study suggest that the verteporfin therapy in the treatment for patients with predominantly classic and classic CNV owing to AMD is encouraging. PMID- 17086168 TI - Loss of nuclear p27 (CDKN1B/KIP1) in colorectal cancer is correlated with microsatellite instability and CIMP. AB - Downregulation of p27 (cyclin-dependent kinase inhibitor-1B, CDKN1B or KIP1) is caused by increased ubiquitin-mediated proteasomal degradation in colorectal cancer, and has been associated with poor prognosis. CpG island methylator phenotype (CIMP) is a phenotype of colorectal cancer with extensive promoter methylation, and associated with high degree of microsatellite instability (MSI H) and BRAF mutations. We have recently shown that both CIMP and MSI-H are inversely associated with downregulation of p21 (CDKN1A or CIP1), another cyclin dependent kinase inhibitor. However, no study to date has examined relationship between p27 and CIMP status in colorectal cancer. Using MethyLight assays, we measured DNA methylation in five CIMP-specific gene promoters {CACNA1G, CDKN2A (p16), CRABP1, MLH1 and NEUROG1} in 706 colorectal cancer samples obtained from two large prospective cohorts. Among the 706 tumors, 112 (16%) were CIMP-high tumors with >or=4/5 methylated promoters. We assessed p27 and p53 expressions by immunohistochemistry. Loss of nuclear p27 expression {observed in 231 tumors (33%)} was significantly associated with CIMP-high, MSI-H and BRAF mutations, and these associations were much more pronounced among p53-negative tumors than p53 positive tumors. When CIMP-high and non-CIMP-high tumors were stratified by MSI status (or KRAS and BRAF status), CIMP-high and MSI-H (but not BRAF mutations) were still significantly associated with nuclear p27 loss. Nuclear p27 loss did not appear to be directly related to CDKN2A (p16) methylation. We conclude that downregulation of nuclear p27 is associated with CIMP-high and MSI-H in colorectal cancer. These associations are stronger among p53 wild-type tumors, implying important interplay of p27 and p53 functions (or dysfunctions) in the development of various molecular subtypes of colorectal cancer. PMID- 17086169 TI - Tunable Frohlich polarons in organic single-crystal transistors. AB - In organic field-effect transistors (FETs), charges move near the surface of an organic semiconductor, at the interface with a dielectric. In the past, the nature of the microscopic motion of charge carriers--which determines the device performance--has been related to the quality of the organic semiconductor. Recently, it was discovered that the nearby dielectric also has an unexpectedly strong influence. The mechanisms responsible for this influence are not understood. To investigate these mechanisms, we have studied transport through organic single-crystal FETs with different gate insulators. We find that the temperature dependence of the mobility evolves from metallic-like to insulating like with increasing dielectric constant of the insulator. The phenomenon is accounted for by a two-dimensional Frohlich polaron model that quantitatively describes our observations and shows that increasing the dielectric polarizability results in a crossover from the weak to the strong polaronic coupling regime. This represents a considerable step forward in our understanding of transport through organic transistors, and identifies a microscopic physical process with a large influence on device performance. PMID- 17086170 TI - Rotation of orbital stripes and the consequent charge-polarized state in bilayer manganites. AB - Nanoscale self-organization of electrons is ubiquitously observed in correlated electron systems such as complex oxides of transition metals. The phenomenon of charge ordering (CO) or the formation of charge stripes, as observed for layered structure cuprates and nickelates, is one such example. Among them, CO in manganites is closely tied to the orbital degree of freedom of 3d electrons, leading to staggered orbital ordering or the formation of orbital stripes in the layered structure. Here, we describe the phenomena of thermally induced rotation of the orbital stripes by 90( composite function) for bilayered manganite crystals with half hole doping, that is, a 1:1 ratio of Mn3+/Mn4+. The rotation of orbital stripes and the consequent CO coupled with the underlying lattice distortion were found to produce the charge-polarized state, as also shown by its optical second-harmonic generation activity. PMID- 17086171 TI - Transformation pathways of silica under high pressure. AB - Network-forming oxides with rigid polyhedral building blocks often possess significant capacity for densification under pressure owing to their open structures. The high-pressure behaviour of these oxides is key to the mechanical properties of engineering materials and geological processes in the Earth's interior. Concurrent molecular-dynamics simulations and first-principles calculations reveal that this densification follows a ubiquitous two-stage mechanism. First, a compact high-symmetry anion sublattice forms, as controlled by strong repulsion between the large oxygen anions, and second, cations redistribute onto the newly created interstices. The same mechanism is observed for two different polymorphs of silica, and in the particular case of cristobalite, is corroborated by the experimental finding of a previously unidentified metastable phase. Our simulations not only clarify the nature of this phase, but also identify its occurrence as key evidence in support of this densification mechanism. PMID- 17086172 TI - BMP-4 is required for hepatic specification of mouse embryonic stem cell-derived definitive endoderm. AB - When differentiated in the presence of activin A in serum-free conditions, mouse embryonic stem cells efficiently generate an endoderm progenitor population defined by the coexpression of either Brachyury, Foxa2 and c-Kit, or c-Kit and Cxcr4. Specification of these progenitors with bone morphogenetic protein-4 in combination with basic fibroblast growth factor and activin A results in the development of hepatic populations highly enriched (45-70%) for cells that express the alpha-fetoprotein and albumin proteins. These cells also express transcripts of Afp, Alb1, Tat, Cps1, Cyp7a1 and Cyp3a11; they secrete albumin, store glycogen, show ultrastructural characteristics of mature hepatocytes, and are able to integrate into and proliferate in injured livers in vivo and mature into hepatocytes expressing dipeptidyl peptidase IV or fumarylacetoacetate hydrolase. Together, these findings establish a developmental pathway in embryonic stem cell differentiation cultures that leads to efficient generation of cells with an immature hepatocytic phenotype. PMID- 17086173 TI - Reversal of mouse hepatic failure using an implanted liver-assist device containing ES cell-derived hepatocytes. AB - Severe acute liver failure, even when transient, must be treated by transplantation and lifelong immune suppression. Treatment could be improved by bioartificial liver (BAL) support, but this approach is hindered by a shortage of human hepatocytes. To generate an alternative source of cells for BAL support, we differentiated mouse embryonic stem (ES) cells into hepatocytes by coculture with a combination of human liver nonparenchymal cell lines and fibroblast growth factor-2, human activin-A and hepatocyte growth factor. Functional hepatocytes were isolated using albumin promoter-based cell sorting. ES cell-derived hepatocytes expressed liver-specific genes, secreted albumin and metabolized ammonia, lidocaine and diazepam. Treatment of 90% hepatectomized mice with a subcutaneously implanted BAL seeded with ES cell-derived hepatocytes or primary hepatocytes improved liver function and prolonged survival, whereas treatment with a BAL seeded with control cells did not. After functioning in the BAL, ES cell-derived hepatocytes developed characteristics nearly identical to those of primary hepatocytes. PMID- 17086174 TI - SUMO-specific protease SUSP4 positively regulates p53 by promoting Mdm2 self ubiquitination. AB - The p53 tumour suppressor has a key role in the control of cell growth and differentiation, and in the maintenance of genome integrity. p53 is kept labile under normal conditions, but in response to stresses, such as DNA damage, it accumulates in the nucleus for induction of cell-cycle arrest, DNA repair or apoptosis. Mdm2 is an ubiquitin ligase that promotes p53 ubiquitination and degradation. Mdm2 is also self-ubiquitinated and degraded. Here, we identified a novel cascade for the increase in p53 level in response to DNA damage. A new SUMO specific protease, SUSP4, removed SUMO-1 from Mdm2 and this desumoylation led to promotion of Mdm2 self-ubiquitination, resulting in p53 stabilization. Moreover, SUSP4 competed with p53 for binding to Mdm2, also resulting in p53 stabilization. Overexpression of SUSP4 inhibited cell growth, whereas knockdown of susp4 by RNA interference (RNAi) promoted of cell growth. UV damage induced SUSP4 expression, leading to an increase in p53 levels in parallel with a decrease in Mdm2 levels. These findings establish a new mechanism for the elevation of cellular p53 levels in response to UV damage. PMID- 17086175 TI - Exo70 interacts with the Arp2/3 complex and regulates cell migration. AB - The exocyst is a multiprotein complex essential for tethering secretory vesicles to specific domains of the plasma membrane for exocytosis. Here, we report that the exocyst component Exo70 interacts with the Arp2/3 complex, a key regulator of actin polymerization. We further show that the exocyst-Arp2/3 interaction is regulated by epidermal growth factor (EGF) signalling. Inhibition of Exo70 by RNA interference (RNAi) or antibody microinjection blocks the formation of actin based membrane protrusions and affects various aspects of cell motility. We propose that Exo70, in addition to functioning in exocytosis, also regulates actin at the leading edges of migrating cells, therefore coordinating cytoskeleton and membrane traffic during cell migration. PMID- 17086176 TI - Key components of the fission machinery are interchangeable. AB - Brefeldin-A ADP-ribosylated substrate (BARS) and dynamin function in membrane fission in distinct intracellular transport pathways, but whether their functions are mechanistically similar is unclear. Here, we show that ARFGAP1, a GTPase activating protein (GAP) for ADP-ribosylation factor 1 (ARF1), couples to either BARS or endophilin B for vesicle formation by the coat protein I (COPI) complex - a finding that reveals an unanticipated mechanistic flexibility in mammalian COPI transport. Because dynamin is coupled to endophilin A in vesicle formation by the clathrin-coat complex, our finding also predicts that dynamin and ARF GAPs are likely to be functional counterparts in membrane fission among different transport pathways that connect intracellular membrane compartments. PMID- 17086177 TI - Natural combinatorial peptide libraries in cyanobacterial symbionts of marine ascidians. AB - A large family of cytotoxic cyclic peptides exemplified by the patellamides has been isolated from ascidians harboring the obligate cyanobacterial symbionts Prochloron spp.. Genome sequence analysis of these symbionts has revealed that Prochloron spp. synthesize patellamides by a ribosomal pathway. To understand how this pathway evolved to produce a suite of related metabolites, we analyzed 46 prochloron-containing ascidians from the tropical Pacific Ocean for the presence of patellamide biosynthetic genes and taxonomic markers. Here, we show that Prochloron spp. generate a diverse library of patellamides using small, hypervariable cassettes within a conserved genetic background. Each symbiont strain contains a single pathway, and mixtures of symbionts within ascidians lead to the accumulation of chemical libraries. We used this information to engineer the production of a new cyclic peptide in Escherichia coli, thereby demonstrating the power of comparative analysis of closely related symbiotic pathways to direct the genetic synthesis of new molecules. PMID- 17086178 TI - A diacylglycerol kinase modulates long-term thermotactic behavioral plasticity in C. elegans. AB - A memory of prior thermal experience governs Caenorhabditis elegans thermotactic behavior. On a spatial thermal gradient, C. elegans tracks isotherms near a remembered temperature we call the thermotactic set-point (T(S)). The T(S) corresponds to the previous cultivation temperature and can be reset by sustained exposure to a new temperature. The mechanisms underlying this behavioral plasticity are unknown, partly because sensory and experience-dependent components of thermotactic behavior have been difficult to separate. Using newly developed quantitative behavioral analyses, we demonstrate that the T(S) represents a weighted average of a worm's temperature history. We identify the DGK-3 diacylglycerol kinase as a thermal memory molecule that regulates the rate of T(S) resetting by modulating the temperature range of synaptic output, but not temperature sensitivity, of the AFD thermosensory neurons. These results provide the first mechanistic insight into the basis of experience-dependent plasticity in this complex behavior. PMID- 17086179 TI - A rigorous experimental framework for detecting protein oligomerization using bioluminescence resonance energy transfer. AB - Bioluminescence resonance energy transfer (BRET), which relies on nonradiative energy transfer between luciferase-coupled donors and GFP-coupled acceptors, is emerging as a useful tool for analyzing the quaternary structures of cell-surface molecules. Conventional BRET analyses are generally done at maximal expression levels and single acceptor/donor ratios. We show that under these conditions substantial energy transfer arises from random interactions within the membrane. The dependence of BRET efficiency on acceptor/donor ratio at fixed surface density, or expression level at a defined acceptor/donor ratio, can nevertheless be used to correctly distinguish between well-characterized monomeric and oligomeric proteins, including a very weak dimer. The pitfalls associated with the nonrigorous treatment of BRET data are illustrated for the case of G protein coupled receptors (GPCRs) proposed to form homophilic and/or mixed oligomers on the basis of previous, conventional BRET experiments. PMID- 17086180 TI - Targeted chromosome elimination from ES-somatic hybrid cells. AB - To engineer a stem cell genome, we developed a technology for targeted elimination of chromosomes from mouse embryonic stem (ES)-somatic hybrid cells. Here we demonstrate the use of a universal chromosome elimination cassette (CEC) for elimination of a single embryonic stem cell (ESC)-derived chromosome 11 or 12, and also both copies of chromosome 6, which harbor pluripotency-associated genes including Nanog. We attribute hybrid-cell pluripotency to the expression of Nanog from the reprogrammed somatic-cell nuclei. PMID- 17086181 TI - CDK1 and calcineurin regulate Maskin association with eIF4E and translational control of cell cycle progression. AB - Maskin regulates assembly of the eIF4F translation initiation complex on messenger RNAs that contain cytoplasmic polyadenylation elements (CPEs) in their 3' untranslated regions. Because Maskin and eIF4G contain similar peptide motifs that bind eIF4E, they compete for occupancy of this factor and consequently control translation. One mRNA that is regulated by Maskin encodes cyclin B1, whose translation oscillates with the early cell cycles of Xenopus laevis embryos. Here we show that Maskin phosphorylation-dephosphorylation also oscillates with the cell cycle and is controlled by the kinase CDK1 and the phosphatase calcineurin. These phosphorylation events control the Maskin-eIF4E interaction and, as a result, translation of cyclin B1 mRNA. Cell cycle progression requires this Maskin-mediated translational regulation. PMID- 17086182 TI - Positional cloning uncovers mutations in PLCE1 responsible for a nephrotic syndrome variant that may be reversible. AB - Nephrotic syndrome, a malfunction of the kidney glomerular filter, leads to proteinuria, edema and, in steroid-resistant nephrotic syndrome, end-stage kidney disease. Using positional cloning, we identified mutations in the phospholipase C epsilon gene (PLCE1) as causing early-onset nephrotic syndrome with end-stage kidney disease. Kidney histology of affected individuals showed diffuse mesangial sclerosis (DMS). Using immunofluorescence, we found PLCepsilon1 expression in developing and mature glomerular podocytes and showed that DMS represents an arrest of normal glomerular development. We identified IQ motif-containing GTPase activating protein 1 as a new interaction partner of PLCepsilon1. Two siblings with a missense mutation in an exon encoding the PLCepsilon1 catalytic domain showed histology characteristic of focal segmental glomerulosclerosis. Notably, two other affected individuals responded to therapy, making this the first report of a molecular cause of nephrotic syndrome that may resolve after therapy. These findings, together with the zebrafish model of human nephrotic syndrome generated by plce1 knockdown, open new inroads into pathophysiology and treatment mechanisms of nephrotic syndrome. PMID- 17086183 TI - A double-switch system regulates male courtship behavior in male and female Drosophila melanogaster. AB - Current models describe male-specific fruitless (fruM) as a genetic 'switch' regulating sexual behavior in Drosophila melanogaster, and they postulate that female (F) and male (M) doublesex (dsx) products control body sexual morphology. In contradiction to this simple model, we show that dsx, as well as fruM and non sex-specific retained (retn), affect both male and female sexual behaviors. In females, both retn and dsxF contribute to female receptivity, and both genes act to repress male-like courtship activity in the presence or absence of fruM. In males, consistent with the opposing functions of dsxM and dsxF, dsxM acts as a positive factor for male courtship. retn also acts counter to fruM in the development of the male-specific muscle of Lawrence. Molecularly, retn seems to regulate sexual behavior via a previously described complex that represses zerknullt. Thus, we show that fru and dsx together act as a 'switch' system regulating behavior in the context of other developmental genes, such as retn. PMID- 17086184 TI - Comparative genome sequencing of Escherichia coli allows observation of bacterial evolution on a laboratory timescale. AB - We applied whole-genome resequencing of Escherichia coli to monitor the acquisition and fixation of mutations that conveyed a selective growth advantage during adaptation to a glycerol-based growth medium. We identified 13 different de novo mutations in five different E. coli strains and monitored their fixation over a 44-d period of adaptation. We obtained proof that the observed spontaneous mutations were responsible for improved fitness by creating single, double and triple site-directed mutants that had growth rates matching those of the evolved strains. The success of this new genome-scale approach indicates that real-time evolution studies will now be practical in a wide variety of contexts. PMID- 17086185 TI - CXorf6 is a causative gene for hypospadias. AB - 46,XY disorders of sex development (DSD) refer to a wide range of abnormal genitalia, including hypospadias, which affects approximately 0.5% of male newborns. We identified three different nonsense mutations of CXorf6 in individuals with hypospadias and found that its mouse homolog was specifically expressed in fetal Sertoli and Leydig cells around the critical period for sex development. These data imply that CXorf6 is a causative gene for hypospadias. PMID- 17086186 TI - Integrin signaling in neutrophils and macrophages uses adaptors containing immunoreceptor tyrosine-based activation motifs. AB - At sites of inflammation, ligation of leukocyte integrins is critical for the activation of cellular effector functions required for host defense. However, the signaling pathways linking integrin ligation to cellular responses are poorly understood. Here we show that integrin signaling in neutrophils and macrophages requires adaptors containing immunoreceptor tyrosine-based activation motifs (ITAMs). Neutrophils and macrophages lacking two ITAM-containing adaptor proteins, DAP12 and FcRgamma, were defective in integrin-mediated responses. Activation of the tyrosine kinase Syk by integrins required that DAP12 and FcRgamma were first phosphorylated by Src family kinases. Retroviral transduction of neutrophils and macrophages with wild-type and mutant Syk or DAP12 demonstrated that the Src homology 2 domains of Syk and the ITAM of DAP12 were required for integrin signaling. Our data show that integrin signaling for the activation of cellular responses in neutrophils and macrophages proceeds by an immunoreceptor-like mechanism. PMID- 17086187 TI - Fever-range thermal stress promotes lymphocyte trafficking across high endothelial venules via an interleukin 6 trans-signaling mechanism. AB - Fever is an evolutionarily conserved response during acute inflammation, although its physiological benefit is poorly understood. Here we show thermal stress in the range of fever temperatures increased the intravascular display of two 'gatekeeper' homing molecules, intercellular adhesion molecule 1 (ICAM-1) and CCL21 chemokine, exclusively in high endothelial venules (HEVs) that are chief portals for the entry of blood-borne lymphocytes into lymphoid organs. Enhanced endothelial expression of ICAM-1 and CCL21 was linked to increased lymphocyte trafficking across HEVs. A bifurcation in the mechanisms controlling HEV adhesion was demonstrated by evidence that the thermal induction of ICAM-1 but not of CCL21 involved an interleukin 6 trans-signaling pathway. Our findings identify the 'HEV axis' as a thermally sensitive alert system that heightens immune surveillance during inflammation by amplifying lymphocyte trafficking to lymphoid organs. PMID- 17086188 TI - Transcriptional regulator Id2 mediates CD8+ T cell immunity. AB - Transcriptional programs that initiate and sustain the proliferation, differentiation and survival of CD8(+) T cells during immune responses are not completely understood. Here we show that inhibitor of DNA binding 2 (Id2), an antagonist of E protein transcription factors, was upregulated in CD8(+) T cells during infection and that expression of Id2 was maintained in memory CD8(+) T cells. Although Id2-deficient naive CD8(+) T cells recognized antigen and proliferated normally early after infection, effector CD8(+) T cells did not accumulate because the cells were highly susceptible to apoptosis. Id2-deficient CD8(+) T cells responding to infection had changes in the expression of genes that influence survival and had altered memory formation. Our data emphasize the importance of Id2 in regulating gene expression by CD8(+) T cells and the magnitude of effector responses, suggesting a mechanism involving Id protein- and E protein-mediated survival and differentiation of mature T cells. PMID- 17086189 TI - Hyperoxia causes angiopoietin 2-mediated acute lung injury and necrotic cell death. AB - The angiogenic growth factor angiopoietin 2 (Ang2) destabilizes blood vessels, enhances vascular leak and induces vascular regression and endothelial cell apoptosis. We considered that Ang2 might be important in hyperoxic acute lung injury (ALI). Here we have characterized the responses in lungs induced by hyperoxia in wild-type and Ang2-/- mice or those given either recombinant Ang2 or short interfering RNA (siRNA) targeted to Ang2. During hyperoxia Ang2 expression is induced in lung epithelial cells, while hyperoxia-induced oxidant injury, cell death, inflammation, permeability alterations and mortality are ameliorated in Ang2-/- and siRNA-treated mice. Hyperoxia induces and activates the extrinsic and mitochondrial cell death pathways and activates initiator and effector caspases through Ang2-dependent pathways in vivo. Ang2 increases inflammation and cell death during hyperoxia in vivo and stimulates epithelial necrosis in hyperoxia in vitro. Ang2 in plasma and alveolar edema fluid is increased in adults with ALI and pulmonary edema. Tracheal Ang2 is also increased in neonates that develop bronchopulmonary dysplasia. Ang2 is thus a mediator of epithelial necrosis with an important role in hyperoxic ALI and pulmonary edema. PMID- 17086190 TI - Quantification of antibody responses against multiple antigens of the two infectious forms of Vaccinia virus provides a benchmark for smallpox vaccination. AB - Smallpox was eradicated without an adequate understanding of how vaccination induced protection. In response to possible bioterrorism with smallpox, the UK government vaccinated approximately 300 health care workers with vaccinia virus (VACV) strain Lister. Antibody responses were analyzed using ELISA for multiple surface antigens of the extracellular enveloped virus (EEV) and the intracellular mature virus (IMV), plaque reduction neutralization and a fluorescence-based flow cytometric neutralization assay. Antibody depletion experiments showed that the EEV surface protein B5 is the only target responsible for EEV neutralization in vaccinated humans, whereas multiple IMV surface proteins, including A27 and H3, are targets for IMV-neutralizing antibodies. These data suggest that it would be unwise to exclude the B5 protein from a future smallpox vaccine. Repeated vaccination provided significantly higher B5-specific and thus EEV-neutralizing antibody responses. These data provide a benchmark against which new, safer smallpox vaccines and residual immunity can be compared. PMID- 17086191 TI - Resveratrol improves health and survival of mice on a high-calorie diet. AB - Resveratrol (3,5,4'-trihydroxystilbene) extends the lifespan of diverse species including Saccharomyces cerevisiae, Caenorhabditis elegans and Drosophila melanogaster. In these organisms, lifespan extension is dependent on Sir2, a conserved deacetylase proposed to underlie the beneficial effects of caloric restriction. Here we show that resveratrol shifts the physiology of middle-aged mice on a high-calorie diet towards that of mice on a standard diet and significantly increases their survival. Resveratrol produces changes associated with longer lifespan, including increased insulin sensitivity, reduced insulin like growth factor-1 (IGF-I) levels, increased AMP-activated protein kinase (AMPK) and peroxisome proliferator-activated receptor-gamma coactivator 1alpha (PGC-1alpha) activity, increased mitochondrial number, and improved motor function. Parametric analysis of gene set enrichment revealed that resveratrol opposed the effects of the high-calorie diet in 144 out of 153 significantly altered pathways. These data show that improving general health in mammals using small molecules is an attainable goal, and point to new approaches for treating obesity-related disorders and diseases of ageing. PMID- 17086192 TI - Crystal structure of a protein phosphatase 2A heterotrimeric holoenzyme. AB - Protein phosphatase 2A (PP2A) is a principal Ser/Thr phosphatase, the deregulation of which is associated with multiple human cancers, Alzheimer's disease and increased susceptibility to pathogen infections. How PP2A is structurally organized and functionally regulated remains unclear. Here we report the crystal structure of an AB'C heterotrimeric PP2A holoenzyme. The structure reveals that the HEAT repeats of the scaffold A subunit form a horseshoe-shaped fold, holding the catalytic C and regulatory B' subunits together on the same side. The regulatory B' subunit forms pseudo-HEAT repeats and interacts with the C subunit near the active site, thereby defining substrate specificity. The methylated carboxy-terminal tail of the C subunit interacts with a highly negatively charged region at the interface between A and B' subunits, suggesting that the C-terminal carboxyl methylation of the C subunit promotes B' subunit recruitment by neutralizing charge repulsion. Together, our structural results establish a crucial foundation for understanding PP2A assembly, substrate recruitment and regulation. PMID- 17086193 TI - Following the signal sequence from ribosomal tunnel exit to signal recognition particle. AB - Membrane and secretory proteins can be co-translationally inserted into or translocated across the membrane. This process is dependent on signal sequence recognition on the ribosome by the signal recognition particle (SRP), which results in targeting of the ribosome-nascent-chain complex to the protein conducting channel at the membrane. Here we present an ensemble of structures at subnanometre resolution, revealing the signal sequence both at the ribosomal tunnel exit and in the bacterial and eukaryotic ribosome-SRP complexes. Molecular details of signal sequence interaction in both prokaryotic and eukaryotic complexes were obtained by fitting high-resolution molecular models. The signal sequence is presented at the ribosomal tunnel exit in an exposed position ready for accommodation in the hydrophobic groove of the rearranged SRP54 M domain. Upon ribosome binding, the SRP54 NG domain also undergoes a conformational rearrangement, priming it for the subsequent docking reaction with the NG domain of the SRP receptor. These findings provide the structural basis for improving our understanding of the early steps of co-translational protein sorting. PMID- 17086194 TI - Klotho converts canonical FGF receptor into a specific receptor for FGF23. AB - FGF23 is a unique member of the fibroblast growth factor (FGF) family because it acts as a hormone that derives from bone and regulates kidney functions, whereas most other family members are thought to regulate various cell functions at a local level. The renotropic activity of circulating FGF23 indicates the possible presence of an FGF23-specific receptor in the kidney. Here we show that a previously undescribed receptor conversion by Klotho, a senescence-related molecule, generates the FGF23 receptor. Using a renal homogenate, we found that Klotho binds to FGF23. Forced expression of Klotho enabled the high-affinity binding of FGF23 to the cell surface and restored the ability of a renal cell line to respond to FGF23 treatment. Moreover, FGF23 incompetence was induced by injecting wild-type mice with an anti-Klotho monoclonal antibody. Thus, Klotho is essential for endogenous FGF23 function. Because Klotho alone seemed to be incapable of intracellular signalling, we searched for other components of the FGF23 receptor and found FGFR1(IIIc), which was directly converted by Klotho into the FGF23 receptor. Thus, the concerted action of Klotho and FGFR1(IIIc) reconstitutes the FGF23 receptor. These findings provide insights into the diversity and specificity of interactions between FGF and FGF receptors. PMID- 17086195 TI - Self-incompatibility in Papaver targets soluble inorganic pyrophosphatases in pollen. AB - In higher plants, sexual reproduction involves interactions between pollen and pistil. A key mechanism to prevent inbreeding is self-incompatibility through rejection of incompatible ('self') pollen. In Papaver rhoeas, S proteins encoded by the stigma interact with incompatible pollen, triggering a Ca2+-dependent signalling network resulting in pollen tube inhibition and programmed cell death. The cytosolic phosphoprotein p26.1, which has been identified in incompatible pollen, shows rapid, self-incompatibility-induced Ca2+-dependent hyperphosphorylation in vivo. Here we show that p26.1 comprises two proteins, Pr p26.1a and Pr-p26.1b, which are soluble inorganic pyrophosphatases (sPPases). These proteins have classic Mg2+-dependent sPPase activity, which is inhibited by Ca2+, and unexpectedly can be phosphorylated in vitro. We show that phosphorylation inhibits sPPase activity, establishing a previously unknown mechanism for regulating eukaryotic sPPases. Reduced sPPase activity is predicted to result in the inhibition of many biosynthetic pathways, suggesting that there may be additional mechanisms of self-incompatibility-mediated pollen tube inhibition. We provide evidence that sPPases are required for growth and that self-incompatibility results in an increase in inorganic pyrophosphate, implying a functional role for Pr-p26.1. PMID- 17086196 TI - Neuroscience: a memory boost while you sleep. PMID- 17086197 TI - Medicine: grapes versus gluttony. PMID- 17086198 TI - In vivo enhancer analysis of human conserved non-coding sequences. AB - Identifying the sequences that direct the spatial and temporal expression of genes and defining their function in vivo remains a significant challenge in the annotation of vertebrate genomes. One major obstacle is the lack of experimentally validated training sets. In this study, we made use of extreme evolutionary sequence conservation as a filter to identify putative gene regulatory elements, and characterized the in vivo enhancer activity of a large group of non-coding elements in the human genome that are conserved in human pufferfish, Takifugu (Fugu) rubripes, or ultraconserved in human-mouse-rat. We tested 167 of these extremely conserved sequences in a transgenic mouse enhancer assay. Here we report that 45% of these sequences functioned reproducibly as tissue-specific enhancers of gene expression at embryonic day 11.5. While directing expression in a broad range of anatomical structures in the embryo, the majority of the 75 enhancers directed expression to various regions of the developing nervous system. We identified sequence signatures enriched in a subset of these elements that targeted forebrain expression, and used these features to rank all approximately 3,100 non-coding elements in the human genome that are conserved between human and Fugu. The testing of the top predictions in transgenic mice resulted in a threefold enrichment for sequences with forebrain enhancer activity. These data dramatically expand the catalogue of human gene enhancers that have been characterized in vivo, and illustrate the utility of such training sets for a variety of biological applications, including decoding the regulatory vocabulary of the human genome. PMID- 17086199 TI - A functional RNAi screen for regulators of receptor tyrosine kinase and ERK signalling. AB - Receptor tyrosine kinase (RTK) signalling through extracellular-signal-regulated kinases (ERKs) has pivotal roles during metazoan development, underlying processes as diverse as fate determination, differentiation, proliferation, survival, migration and growth. Abnormal RTK/ERK signalling has been extensively documented to contribute to developmental disorders and disease, most notably in oncogenic transformation by mutant RTKs or downstream pathway components such as Ras and Raf. Although the core RTK/ERK signalling cassette has been characterized by decades of research using mammalian cell culture and forward genetic screens in model organisms, signal propagation through this pathway is probably regulated by a larger network of moderate, context-specific proteins. The genes encoding these proteins may not have been discovered through traditional screens owing, in particular, to the requirement for visible phenotypes. To obtain a global view of RTK/ERK signalling, we performed an unbiased, RNA interference (RNAi), genome wide, high-throughput screen in Drosophila cells using a novel, quantitative, cellular assay monitoring ERK activation. Here we show that ERK pathway output integrates a wide array of conserved cellular processes. Further analysis of selected components-in multiple cell types with different RTK ligands and oncogenic stimuli-validates and classifies 331 pathway regulators. The relevance of these genes is highlighted by our isolation of a Ste20-like kinase and a PPM family phosphatase that seem to regulate RTK/ERK signalling in vivo and in mammalian cells. Novel regulators that modulate specific pathway outputs may be selective targets for drug discovery. PMID- 17086200 TI - Boosting slow oscillations during sleep potentiates memory. AB - There is compelling evidence that sleep contributes to the long-term consolidation of new memories. This function of sleep has been linked to slow (<1 Hz) potential oscillations, which predominantly arise from the prefrontal neocortex and characterize slow wave sleep. However, oscillations in brain potentials are commonly considered to be mere epiphenomena that reflect synchronized activity arising from neuronal networks, which links the membrane and synaptic processes of these neurons in time. Whether brain potentials and their extracellular equivalent have any physiological meaning per se is unclear, but can easily be investigated by inducing the extracellular oscillating potential fields of interest. Here we show that inducing slow oscillation-like potential fields by transcranial application of oscillating potentials (0.75 Hz) during early nocturnal non-rapid-eye-movement sleep, that is, a period of emerging slow wave sleep, enhances the retention of hippocampus-dependent declarative memories in healthy humans. The slowly oscillating potential stimulation induced an immediate increase in slow wave sleep, endogenous cortical slow oscillations and slow spindle activity in the frontal cortex. Brain stimulation with oscillations at 5 Hz--another frequency band that normally predominates during rapid-eye-movement sleep--decreased slow oscillations and left declarative memory unchanged. Our findings indicate that endogenous slow potential oscillations have a causal role in the sleep-associated consolidation of memory, and that this role is enhanced by field effects in cortical extracellular space. PMID- 17086201 TI - Thymic selection threshold defined by compartmentalization of Ras/MAPK signalling. AB - A healthy individual can mount an immune response to exogenous pathogens while avoiding an autoimmune attack on normal tissues. The ability to distinguish between self and non-self is called 'immunological tolerance' and, for T lymphocytes, involves the generation of a diverse pool of functional T cells through positive selection and the removal of overtly self-reactive thymocytes by negative selection during T-cell ontogeny. To elucidate how thymocytes arrive at these cell fate decisions, here we have identified ligands that define an extremely narrow gap spanning the threshold that distinguishes positive from negative selection. We show that, at the selection threshold, a small increase in ligand affinity for the T-cell antigen receptor leads to a marked change in the activation and subcellular localization of Ras and mitogen-activated protein kinase (MAPK) signalling intermediates and the induction of negative selection. The ability to compartmentalize signalling molecules differentially in the cell endows the thymocyte with the ability to convert a small change in analogue input (affinity) into a digital output (positive versus negative selection) and provides the basis for establishing central tolerance. PMID- 17086202 TI - Wza the translocon for E. coli capsular polysaccharides defines a new class of membrane protein. AB - Many types of bacteria produce extracellular polysaccharides (EPSs). Some are secreted polymers and show only limited association with the cell surface, whereas others are firmly attached to the cell surface and form a discrete structural layer, the capsule, which envelopes the cell and allows the bacteria to evade or counteract the host immune system. EPSs have critical roles in bacterial colonization of surfaces, such as epithelia and medical implants; in addition some EPSs have important industrial and biomedical applications in their own right. Here we describe the 2.26 A resolution structure of the 340 kDa octamer of Wza, an integral outer membrane lipoprotein, which is essential for group 1 capsule export in Escherichia coli. The transmembrane region is a novel alpha-helical barrel. The bulk of the Wza structure is located in the periplasm and comprises three novel domains forming a large central cavity. Wza is open to the extracellular environment but closed to the periplasm. We propose a route and mechanism for translocation of the capsular polysaccharide. This work may provide insight into the export of other large polar molecules such as DNA and proteins. PMID- 17086203 TI - Boc is a receptor for sonic hedgehog in the guidance of commissural axons. AB - In the spinal cord, sonic hedgehog (Shh) is secreted by the floor plate to control the generation of distinct classes of ventral neurons along the dorsoventral axis. Genetic and in vitro studies have shown that Shh also later acts as a midline-derived chemoattractant for commissural axons. However, the receptor(s) responsible for Shh attraction remain unknown. Here we show that two Robo-related proteins, Boc and Cdon, bind specifically to Shh and are therefore candidate receptors for the action of Shh as an axon guidance ligand. Boc is expressed by commissural neurons, and targeted disruption of Boc in mouse results in the misguidance of commissural axons towards the floor plate. RNA-interference mediated knockdown of Boc impairs the ability of rat commissural axons to turn towards an ectopic source of Shh in vitro. Taken together, these data suggest that Boc is essential as a receptor for Shh in commissural axon guidance. PMID- 17086205 TI - Structure of the E. coli signal recognition particle bound to a translating ribosome. AB - The prokaryotic signal recognition particle (SRP) targets membrane proteins into the inner membrane. It binds translating ribosomes and screens the emerging nascent chain for a hydrophobic signal sequence, such as the transmembrane helix of inner membrane proteins. If such a sequence emerges, the SRP binds tightly, allowing the SRP receptor to lock on. This assembly delivers the ribosome-nascent chain complex to the protein translocation machinery in the membrane. Using cryo electron microscopy and single-particle reconstruction, we obtained a 16 A structure of the Escherichia coli SRP in complex with a translating E. coli ribosome containing a nascent chain with a transmembrane helix anchor. We also obtained structural information on the SRP bound to an empty E. coli ribosome. The latter might share characteristics with a scanning SRP complex, whereas the former represents the next step: the targeting complex ready for receptor binding. High-resolution structures of the bacterial ribosome and of the bacterial SRP components are available, and their fitting explains our electron microscopic density. The structures reveal the regions that are involved in complex formation, provide insight into the conformation of the SRP on the ribosome and indicate the conformational changes that accompany high-affinity SRP binding to ribosome nascent chain complexes upon recognition of the signal sequence. PMID- 17086206 TI - Differential role for very late antigen-5 in mobilization and homing of hematopoietic stem cells. AB - The role of very late antigen-5 (VLA-5) in homing and mobilization of hematopoietic stem cells from normal bone marrow (NBM) and bone marrow (MBM) and peripheral blood (MPB) from mobilized mice was investigated. We found a decreased number of VLA-5-expressing cells in the lineage-negative fraction of MPB. However, virtually all stem/progenitor cells were present in the VLA-5(+) fraction and hence mobilization of hematopoietic stem cell subsets does not coincide with a downregulation of VLA-5. Stem/progenitor cells from MPB and MBM demonstrated enhanced stromal-derived factor-alpha-induced migration. This enhanced migration correlates with an improved hematopoietic reconstitution potential, with the migrated MPB cells showing the fastest reconstitution. Interestingly, homing of MPB, MBM and NBM stem/progenitor cells in bone marrow and spleen did not differ and is therefore not responsible for the differences in hematopoietic reconstitution. The observed increase in VLA-5(+) cells in the recipients after transplantation can most probably be attributed to selective homing of VLA-5(+) cells instead of an upregulation of VLA-5. Treatment with an antibody to VLA-5 partially inhibited bone marrow homing of progenitor cells, whereas homing in the spleen was hardly affected. These data indicate a differential role for VLA-5 in the movement of stem cells from and toward bone marrow. PMID- 17086204 TI - Global trends of whole-genome duplications revealed by the ciliate Paramecium tetraurelia. AB - The duplication of entire genomes has long been recognized as having great potential for evolutionary novelties, but the mechanisms underlying their resolution through gene loss are poorly understood. Here we show that in the unicellular eukaryote Paramecium tetraurelia, a ciliate, most of the nearly 40,000 genes arose through at least three successive whole-genome duplications. Phylogenetic analysis indicates that the most recent duplication coincides with an explosion of speciation events that gave rise to the P. aurelia complex of 15 sibling species. We observed that gene loss occurs over a long timescale, not as an initial massive event. Genes from the same metabolic pathway or protein complex have common patterns of gene loss, and highly expressed genes are over retained after all duplications. The conclusion of this analysis is that many genes are maintained after whole-genome duplication not because of functional innovation but because of gene dosage constraints. PMID- 17086207 TI - Engraftment and survival following hematopoietic stem cell transplantation for osteopetrosis using a reduced intensity conditioning regimen. AB - Autosomal recessive osteopetrosis (OP) is a disease characterized by osteoclast dysfunction, leading to multisystem morbidity and death of most affected children. Hematopoietic stem cell transplantation (HSCT) is the treatment of choice for OP, but this patient population is particularly prone to post transplant complications and death after myeloablative conditioning. To determine the potential of achieving improved overall outcomes in these patients by decreasing pre-transplant mortality, we investigated engraftment and survival following a reduced intensity regimen including busulfan, fludarabine and total lymphoid irradiation. We report outcomes in 11 patients. All six patients who received a bone marrow or peripheral stem cell graft engrafted with >75% donor chimerism. In contrast, all five recipients of unrelated cord blood as a stem cell source for a first graft failed to demonstrate donor hematopoietic chimerism. The day 100 and 6-month mortality was low at 9%. One year after HSCT, six of 11 patients (55%) were surviving. Our data suggest that this regimen results in low peri-transplant mortality without compromising engraftment when a marrow or peripheral stem cell graft is used. An umbilical cord blood graft, however, should be used with caution for patients with OP when this or a similar reduced intensity regimen is used. PMID- 17086208 TI - Ikaros is a mutational target for lymphomagenesis in Mlh1-deficient mice. AB - Deficiencies in DNA mismatch repair (MMR) result in replication errors within key tumor suppressor genes or oncogenes, and cause hereditary nonpolyposis colorectal cancer (HNPCC). Hematological malignancy with microsatellite instability is also associated with defective MMR, but little is known about the target genes for MMR. Here we identified Ikaros, a master transcription factor of lymphoid lineage commitment and differentiation, as a mutational target in spontaneous and radiation-induced T-cell lymphomas in Mlh1-deficient mice. Three quarters of lymphomas lacked Ikaros protein expression, which resulted from a frameshift mutation that created a stop codon. Mononucleotide repeat sequences at 1029 1034(C)6 and 1567-1572(G)6 in Ikaros were mutational hot spots with a one-base deletion occurring with a frequency of 45 and 50%, respectively. Point mutations and splicing alterations were also observed. In total, 85% of the lymphomas showed aberrations in Ikaros. The characteristic of Mlh1-deficient lymphomas is harboring of multiple mutations simultaneously in the same tumor, displaying a combination of two frameshift mutations at different repeats, frameshift and point mutations, and/or deletion mutations. This is the first report of Ikaros mutations coupled with Mlh1 deficiency in lymphomagenesis. PMID- 17086209 TI - Identifying candidate colon cancer tumor suppressor genes using inhibition of nonsense-mediated mRNA decay in colon cancer cells. AB - Inhibition of the nonsense-mediated decay (NMD) mechanism in cells results in stabilization of transcripts carrying premature translation termination codons. A strategy referred to as gene identification by NMD inhibition (GINI) has been proposed to identify genes carrying nonsense mutations. Genes containing frameshift mutations in colon cancer cell line have been identified using a modified version of GINI. To increase the efficiency of identifying mutant genes using GINI, we have now further improved the strategy. In this approach, inhibition of NMD with emetine is complemented with inhibiting NMD by blocking the phosphorylation of the hUpf1 protein with caffeine. In addition, to enhance the GINI strategy, comparing mRNA level alterations produced by inhibiting transcription alone or inhibiting transcription together with NMD following caffeine pretreatment were used for the efficient identification of false positives produced as a result of stress response to NMD inhibition. To demonstrate the improved efficiency of this approach, we analysed colon cancer cell lines showing microsatellite instability. Bi-allelic inactivating mutations were found in the FXR1, SEC31L1, NCOR1, BAT3, PHF14, ZNF294, C19ORF5 genes as well as genes coding for proteins with yet unknown functions. PMID- 17086210 TI - Identification of multiple SNT-binding sites on NPM-ALK oncoprotein and their involvement in cell transformation. AB - The t(2;5) chromosomal translocation occurs in anaplastic large-cell lymphoma arising from activated T lymphocytes. This genomic rearrangement generates the nucleophosmin (NPM)-anaplastic lymphoma kinase (ALK) oncoprotein that is a chimeric protein consisting of parts of the nuclear protein NPM and ALK receptor protein-tyrosine kinase. We used yeast two-hybrid screening to identify an adaptor protein Suc1-associated neurotrophic factor-induced tyrosine phosphorylated target (SNT)-2 as a new partner that interacted with the cytoplasmic domain of ALK. Immunoprecipitation assay revealed that SNT-1 and SNT 2 interacted with NPM-ALK and kinase-negative NPM-ALK mutant. Y156, Y567 and a 19 amino-acid sequence (aa 631-649) of NPM-ALK were essential for this interaction. The interaction through Y156 and Y567 was dependent on phosphorylation of these tyrosines, whereas the interaction through the 19-amino-acid sequence was independent of phosphorylation. NPM-ALK mutant protein mutated at these three binding sites showed significantly reduced transforming activity. This transformation-defective NPM-ALK mutant still interacted with signal transducing proteins such as phospholipase C-gamma and phosphatidylinositol 3-kinase, which were previously reported to be relevant to NPM-ALK-dependent tumorigenesis. These observations indicate that the three SNT-binding sites of NPM-ALK are important for its transforming activity. This raises a possibility that SNT family proteins play significant roles in cellular transformation triggered by NPM-ALK, which though remains to be verified. PMID- 17086211 TI - Suppression of hypoxic cell death by APIP-induced sustained activation of AKT and ERK1/2. AB - Apaf-1-interacting protein (APIP) was previously isolated as an inhibitor of mitochondrial cell death interacting with Apaf-1. Here, we report a hypoxia selective antiapoptotic activity of APIP that induces the activation of AKT and extracellular signal-regulated kinase (ERK)1/2. Stable expression of APIP in C2C12 (C2C12/APIP) cells suppressed cell death induced by hypoxia and etoposide. Unlike etoposide, however, APIP induces the sustained activation of AKT and ERK1/2 and the phosphorylation of caspase-9 during hypoxia. Inhibition of AKT and ERK1/2 activation by the treatments with phosphatidylinositol 3'-kinase and mitogen-activated protein kinase kinase (MEK)1/2 inhibitors sensitized C2C12/APIP cells to hypoxic cell death and abolished the hypoxia-induced phosphorylation of caspase-9. Further, overexpression of phosphorylation-mimic caspase-9 mutants (caspase-9-T125E and caspase-9-S196D), but not phosphorylation-defective caspase 9 mutants (caspase-9-T125A and caspase-9-S196A), effectively suppressed hypoxia induced death of C2C12 cells. These results elucidate a novel Apaf-1-independent antiapoptotic activity of APIP during hypoxic cell death, inducing the sustained activation of AKT and ERK1/2 and leading to caspase-9 phosphorylation. PMID- 17086212 TI - Drug-induced expression of the cellular adhesion molecule L1CAM confers anti apoptotic protection and chemoresistance in pancreatic ductal adenocarcinoma cells. AB - Pancreatic ductal adenocarcinoma (PDAC) is characterized by rapid tumor progression, high metastatic potential and profound chemoresistance. We recently reported that induction of a chemoresistant phenotype in the PDAC cell line PT45 P1 by long-term chemotherapy involves an increased interleukin 1 beta (IL1beta) dependent secretion of nitric oxide (NO) accounting for efficient caspase inhibition. In the present study, we elucidated the involvement of L1CAM, an adhesion molecule previously found in other malignancies, in this NO-dependent chemoresistance. Chemoresistant PT45-P1res cells, but not chemosensitive parental PT45-P1 cells, express high levels of L1CAM in an ILbeta-dependent fashion. PT45 P1res cells subjected to short interfering RNA (siRNA)-mediated L1CAM knock-down exhibited reduced inducible nitric oxide synthase expression and NO secretion, as well as a significant increase of anti-cancer drug-induced caspase activation, an effect reversed by the NO donor S-nitroso-N-acetyl-D,L-penicillamine. Conversely, overexpression of L1CAM in PT45-P1 cells conferred anti-apoptotic protection to anti-cancer drug treatment. Interestingly, L1CAM ectodomain shedding, in example, by ADAM10, as reported for other L1CAM-related activities, seemed to be dispensable for anti-apoptotic protection by L1CAM. Neither the shedded L1CAM ectodomain was detected in chemoresistant L1CAM-expressing PT45-P1 cells nor did the administration of various metalloproteinase inhibitors affect L1CAM-dependent chemoresistance. Immunohistochemical analysis revealed L1CAM expression in 80% of pancreatic cancer specimens, supporting a potential role of L1CAM in the malignancy of this tumor. These findings substantiate our understanding of the molecular mechanisms leading to chemoresistance in PDAC cells and indicate the importance of L1CAM in this scenario. PMID- 17086214 TI - [Overview and insight]. PMID- 17086213 TI - Unr, a cytoplasmic RNA-binding protein with cold-shock domains, is involved in control of apoptosis in ES and HuH7 cells. AB - Unr (upstream of N-ras) is a cytoplasmic RNA-binding protein involved in the regulation of messenger RNA stability and internal initiation of translation. We have used Unr-deficient murine embryonic stem (ES) cells to analyse Unr role in cell proliferation and response to stress. Disruption of both unr gene copies had no effect on ES cell proliferation. However, after ionizing radiation (IR), clonogenic survival of unr(-/-) ES cells was approximately 3-fold enhanced as compared to unr(+/+) cells. We further determined that IR-induced apoptosis was decreased in unr(-/-) ES cells, and that reintroduction of the unr gene in unr(-/ ) cells restored normal IR-induced apoptosis. Three pro-apoptotic genes, p53, caspase-3 and Gadd45gamma, were downregulated in unr(-/-) ES cells, indicating that Unr, as other cytoplasmic RNA-binding proteins, regulates a complex genetic program, promoting cell death after IR. In contrast, in the human hepatoma cell line HuH7, Unr knockdown using unr-specific small interfering RNAs induced apoptosis, both in untreated and gamma-irradiated cells. Thus, our results establish that Unr acts as a positive or negative regulator of cell death, depending on the cell type. Manipulating the level of Unr may constitute a specific approach to sensitize cancer cells to anticancer treatments. PMID- 17086215 TI - [Does alpha-linolenic acid prevent cardiac death?]. PMID- 17086216 TI - [A renaissance for home hemodialysis?]. PMID- 17086217 TI - [Assessment of community mental health centres by general practitioners]. AB - BACKGROUND: General practitioners' evaluation of Community Mental Health Centres (CMHC) has become a part of the Norwegian system of quality indicators. On the basis of a national survey, we present the way in which GPs have assessed the quality of 77 CMHCs. MATERIAL AND METHODS: A questionnaire was posted to every GP in Norway from November 2005 to March 2006. This was followed up with 2-3 reminders and telephone interviews. 2,415 out of 3,704 GPs (65%) responded. RESULTS: GPs gave highest score for professional competence, the average being 55 on a scale of 0-100, where 100 is best. They gave lowest score for counselling (30) and staff situation (45). The average score given to the various CMHCs varied a lot for all dimensions, the largest difference being for counselling, with the highest average 58 and the lowest 8. There were many significant differences between various CMHCs' scores and the national average. When it came to counselling, the average score for 20 CMHCs was significantly different from that for the other CMHCs. INTERPRETATION: As GPs are important collaborators and users of the CMHCs, their views are valuable. The present findings need to be discussed. Many of the community mental health centres received all in all positive feedback from the GPs, suggesting that they may have useful experiences to share. PMID- 17086218 TI - [Alpha-linolenic acid, cardiovascular disease and sudden death]. AB - BACKGROUND: Several animal experiments have documented that omega-3 fatty acid alpha-linolenic acid (ALA) (C 18:3, n-3) from vegetable oils has beneficial electrophysiological and antiarrhythmic effects. This may explain the protective effect of ALA against cardiovascular diseases and sudden death. MATERIALS AND METHODS: We have continuously and systematically collected and evaluated relevant literature (observational and secondary prophylactic studies) and here present an overview of the effects of ALA on cardiovascular diseases and sudden death. RESULTS: Several observational studies in healthy individuals and in patients with coronary heart disease have indicated that a diet rich in ALA reduces coronary death and the risk for sudden death. A cross-sectional study has shown a beneficial effect of ALA on the progression of coronary atherosclerosis. Three secondary studies have also indicated beneficial effects of ALA on several cardiovascular end-points. Diet studies do have limitations, and the secondary prophylactic studies have not given a definite answer to whether ALA possesses beneficial cardiovascular effects. INTERPRETATION: On the whole, the observational and secondary prophylactic studies indicate that ALA may have cardioprotective effects in healthy individuals and in patients with coronary heart disease. For those who seldom eat fish and only take small amounts of fish oils, a diet rich on ALA may be an alternative. PMID- 17086219 TI - [Home hemodialysis as an alternative to in-center dialysis]. AB - BACKGROUND: Dialysis is a life-sustaining treatment for renal disease in its final stage. The number of patients in dialysis has more than doubled over the last 10 years in Norway. The prognoses still show an increasing number of patients in need of dialysis. As haemodialysis (HD) is complicated to perform at home, peritoneal dialysis is the preferred alternative. HD is usually carried out 3 times a week at hospitals, in satellites or self-dialysis in some other institution. Home HD is rarely used in Norway, as opposed to in other Western European countries. MATERIAL AND METHOD: We describe our first patient offered HD at home and give an overview of home HD. RESULTS AND INTERPRETATION: Empirically, the increased dialysis time in home HD generates positive physical, mental and social effects. The potential of economic savings are considerable. Home HD is a cost-effective method to improve health and solve some of the problems associated with centre dialysis. The treatment generates great demands for equipment and training, and is suitable only for carefully selected patients. Home HD offers considerable more freedom and possibility for more frequent and longer dialysis than centre dialysis. PMID- 17086220 TI - [Treatment of hormone-resistant prostate cancer]. AB - Hormone-resistant prostate cancer is defined by disease progression despite of castration levels of serum testosterone. Due to a good palliative effect and low toxicity, radiotherapy is the cornerstone in treatment of painful bone metastases, the most frequent symptom of advanced hormone-resistant prostate cancer. Patients presenting with spinal cord compression should be assessed for decompressive surgery before radiotherapy. With local growth of prostate cancer and problems with urination, a transurethral resection may palliate symptoms. Postoperative radiotherapy should however be considered for these patients, so local regrowth of the tumour can be prevented. Taxan-based chemotherapy (docetaxel) is the first treatment shown to increase overall survival in patients with hormone-resistant prostate cancer, and is today's standard treatment of Norwegian patients (guidelines from the Norwegian Urological Cancer Group). Secondary hormone treatment and administration of bisphosphonates are other established alternatives for palliation of symptoms in patients with hormone resistant prostate cancer. The survival of future patients with such cancer is expected to improve if multidisciplinary health care teams with knowledge of prostate cancer tumour biology administer new treatments. PMID- 17086221 TI - [Antiplatelet therapy after aspirin-induced upper gastrointestinal bleeding]. AB - It is common practice to replace aspirin with clopidogrel in patients with gastrointestinal intolerance to aspirin. Recent studies suggest that a combination of aspirin and a proton pump inhibitor is a better alternative for these patients. The CAPRIE and CURE studies have not shown any clinically relevant difference in effect between aspirin and clopidogrel. The incidence of bleeding is also similar when aspirin is used in doses < 160 mg. A recent study by Chan et al. concluded that a combination of aspirin and esomeprazole is superior to clopidogrel in the prevention of recurrent gastrointestinal bleeding. Using aspirin and a proton pump inhibitor is also the least expensive alternative. PMID- 17086222 TI - [Travel vaccination]. AB - Vaccination gives good and long-lasting protection against a number of infectious diseases, and is nearly always recommended before journeys to countries with high risk of contagious diseases. It is important to both renew childhood vaccines and to inoculate against hepatitis A. Vaccines against yellow fever and meningococcal disease are compulsory before entering certain countries. Vaccination against hepatitis B, rabies, typhoid fever and perhaps cholera should be considered for persons spending longer periods in endemic areas. There is a risk of Japanese encephalitis in large parts of South-East Asia, and tick-borne encephalitis in certain areas of Europe. PMID- 17086223 TI - [Vaccination against influenza--why, for whom and with which vaccine?]. AB - Every year, 5-10% of Norwegians become sick from influenza. Some groups, such as the elderly and the seriously ill, are at greater risk of contracting a serious form of influenza leading to complications and death. Annual influenza vaccination in such groups reduces the risk for pneumonia, hospitalisation and death. Vaccination of health care workers reduces the mortality rate among patients during the winter season. In Norway, around 900,000 persons belong to a group at risk and should be offered vaccination. In the winter of 2005/06, about 45% of these groups were vaccinated; the aim is 75% by 2010. Whether people get vaccinated or not depends much on their self-perceived need of vaccination. Health care workers play an important role in raising awareness of and spreading information about influenza and vaccination. Sending personal invitations and arranging mass vaccination days are effective in increasing the rate of vaccination. PMID- 17086224 TI - [The development of mucosal vaccines]. AB - The live oral polio vaccine was the first mucosal vaccine accepted for general use. Since then, similar vaccines have been developed against typhoid fever, cholera and rotavirus infection, and a nasal vaccine against influenza has recently been registered in the USA. The only non-living mucosal vaccine on the market today is an oral cholera vaccine consisting of inactivated Vibrio cholerae and the B subunit of cholera toxin. Several groups of scientists are at present working on the development of other mucosal vaccines based on inactivated microbes or parts of them. Results from animal trials at the Norwegian Institute of Public Health, suggest that non-living nasal vaccines can provide protective immunity and may be combined with the same types of vaccines for injection. Clinical trials with nasal vaccines consisting of beta-propiolactone inactivated influenza particles, showed that it was possible to achieve serum concentrations of antibodies at levels providing protection against influenza. IgA antibodies, which were formed in nasal secretions, were specifically aimed at influenza and ought to hinder the spread of the disease. By optimizing the immunization regimes so that the immunological memory is better exploited, and by adding adjuvants to the formulations, it is probable that non-living mucosal vaccines can be realistic alternatives to several of the vaccines now given by injection. PMID- 17086225 TI - [The good quality indicators]. PMID- 17086226 TI - [District psychiatry centers with unsatisfactory quality]. PMID- 17086227 TI - [Weak links in the therapeutic chain]. PMID- 17086228 TI - [Do Norwegian cancer patients receive the care they deserve?]. PMID- 17086229 TI - [New drug financing model causes concerns]. PMID- 17086231 TI - [Confusing advice from the Legeforeningens forskningsinstitut]. PMID- 17086232 TI - [Syncope in children and young adults]. PMID- 17086234 TI - [Abortion--no longer need for research?]. PMID- 17086235 TI - [A life-saving booklet]. PMID- 17086238 TI - [Psychological activity is also important]. PMID- 17086240 TI - [Peace and health]. PMID- 17086241 TI - [Lactate function in the brain]. PMID- 17086242 TI - [Norwegian health and EU research program]. PMID- 17086244 TI - [Histoplasmosis among travellers to Central America]. AB - BACKGROUND: In April 2004, the Norwegian Institute of Public Health was notified about an outbreak of a respiratory illness among participants on a cultural historical theme tour to Central America. One patient was examined in a Swedish hospital where the clinician suspected histoplasmosis, which later was verified by serology and by culture of Histoplasma capsulatum. In order to describe the outbreak and identify the possible source of infection, a clinical and epidemiological study was conducted. MATERIAL AND METHODS: A questionnaire on symptoms, treatment and participation in various activities was sent to all the tour participants. Reminders were sent by e-mail. In addition, some participants underwent a more detailed clinical examination. RESULTS: 19 of the 21 travellers completed the questionnaire. All travellers had visited several caves inhabited by bats in El Salvador and Guatemala. During the last 10 days of the journey, 16 of the respondents (84%) acquired a respiratory illness that lasted for more than 3 weeks. The diagnosis was confirmed by serology in 8 of the 14 tested cases. Antibacterial therapy was given to 11 patients and antifungal therapy to 3 patients. Only one patient was hospitalised. INTERPRETATION: The described outbreak of histoplasmosis was probably caused by exposure to Histoplasma capsulatum during a visit in bat-infested caves. Due to the increasing popularity of adventure and theme travels, this disease may become more frequent in our population. The condition should be considered in travellers with prolonged respiratory illness after visits to caves or other closed environments with little circulation of air that are potentially contaminated with bat or bird droppings. Persons with immunosuppression may develop severe illness, and should consider avoiding places with increased risk. PMID- 17086246 TI - Generation of inhomogeneously polarized laser beams by use of a Sagnac interferometer. AB - A principal scheme for an external cavity technique for changing the polarization of a laser beam based on a modified Sagnac interferometer is proposed. The modified Sagnac interferometer includes standard optical components: a displacement polarizing beam splitter, an angle reflector, and a Dove prism. The radially polarized beams, obtained with the help of the developed scheme, allow the generation of a longitudinally polarized electric field by sharp focusing. The phase correction of radially polarized modes of higher orders leads to increasing the longitudinal field in the focus of the beam. PMID- 17086247 TI - Modeling and optimization for a prismatic snapshot imaging polarimeter. AB - Thin birefringent prisms placed near an image plane introduce sinusoidal fringes onto a 2D polarized scene making possible a snapshot imaging polarimeter, which encodes polarization information into the modulation of the fringes. This approach was introduced by Oka and Kaneko [Opt. Express 11, 1510 (2003)], who analyzed the instrument through the Mueller calculus. We show that the plane-wave assumption adopted in the Mueller theory can introduce unnecessary error in a polarimeter design. To directly take prism effects such as beam splitting and deviating into accounts we introduce a geometric imaging model, which allows for a versatile simulation of the birefringent prisms and provides a means for optimization. A calcite visible system is investigated as an example, which essentially shows how each design parameter affects the overall image quality and how to modify the polarimeter design to optimize overall performance. The approach is applicable to any prismatic imaging polarimeter with different prism materials and different working wavelengths. PMID- 17086248 TI - Scanning transmission microscopy using a position-sensitive detector. AB - Optical data manipulation technologies increasingly employ densely aperiodic optical 3D phase elements. Refinement of such technologies will require the capability to quantitatively characterize the volumetric dielectric modulation of an optical sample to a high level of precision and spatial resolution. We present a scanning transmission microscopy system that uses a position-sensitive detector to impart sensitivity to both the phase and absorption components. We describe the layout of the instrument and then derive its phase and absorption transfer functions. Simulations and experiments are presented to validate the analysis. For phase detection, the instrument possesses depth-sectioning properties similar to those of a confocal microscope without the use of a pinhole, enabling full 3D object reconstruction. PMID- 17086249 TI - Generalization of the Rouard method to an absorbing thin-film stack and application to surface plasmon resonance. AB - In the context of surface plasmon resonance (SPR) kinetic biochips, it is important to model the SPR phenomenon (i.e., extinction of reflectivity) toward biochip design and optimization. The Rouard approach that models reflectivity off a thin-film stack is shown to be extendable to any number of absorbing layers with no added complexity. Using the generalized Rouard method, the effect of SPR is simulated as a function of the wavelength for various metal thicknesses. Given an optimal metal thickness, the dependence of SPR on the angle of incidence and wavelength is also demonstrated. Such a model constitutes a potential basis for the efficient design and optimization of multidimensional sensors. PMID- 17086250 TI - Fabrication of multilayered photochromic memory media using pressure-sensitive adhesives. AB - We fabricated a multilayered medium by a laminating process with pressure sensitive adhesives. It was possible to reduce the thickness variations of both photosensitive layers and transparent layers by applying laminating films. This method is easy to use to pile up many layers for a multilayered medium. We fabricated twenty recording layers and demonstrated the capability to record with significant reading results. The recorded data in each layer were read out without crosstalk. We evaluated the signal-to-noise ratio and crosstalk between neighboring layers. It was found that the signal-to-noise ratio of a multilayered medium was higher than 50 dB. PMID- 17086251 TI - Polarimeter with two ferroelectric liquid-crystal modulators attached to the Yunnan solar tower. AB - A polarimeter to be mounted on the Yunnan solar tower is described. It features the ability to simultaneously measure the magnetic fields of the solar photosphere and chromosphere by analyzing the Stokes spectra of those magnetosensitive lines forming in the two regions with very high efficiency of polarization measurement. The polarimeter consists of two ferroelectric liquid crystals and one lambda/4 wave plate before a polarizing beam splitter. The achromatism of the design is emphasized to get the maximum combination efficiency over a spectral range from 5000 to 6000 A. For the used solar absorption lines MgI517.27, FeI525.06, FeI630.15, and FeI630.25 nm, the design gives theoretical efficiencies of polarization measurements, which are 0.999, 1.0, 0.943, and 0.943, respectively. A comparison with other reference polarimeters, such as the Synoptic Optical Long-term Investigation of the Sun, the Tenerife infrared polarimeter, and the La Palma Stokes Polarimeter, is carried out. PMID- 17086252 TI - Affine cryptosystem of double-random-phase encryption based on the fractional Fourier transform. AB - An affine mapping mathematical expression of the double-random-phase encryption technique has been deduced utilizing the matrix form of discrete fractional Fourier transforms. This expression clearly describes the encryption laws of the double-random-phase encoding techniques based on both the fractional Fourier transform and the ordinary Fourier transform. The encryption process may be regarded as a substantial optical realization of the affine cryptosystem. It has been illustrated that the encryption process converts the original image into a white Gaussian noise with a zero-mean value. Also, the decryption process converts the data deviations of the encrypted image into white Gaussian noises, regardless of the type of data deviations. These noises superimpose on the decrypted image and degrade the signal-to-noise ratio. Numerical simulations have been implemented for the different types of noises introduced into the encrypted image, such as the white noise with uniform distribution probability, the white noise with Gaussian distribution probability, colored noise, and the partial occlusion of the encrypted image. PMID- 17086253 TI - Diffractive optical elements for beam shaping of monochromatic spatially incoherent light. AB - Fresnel-type diffractive optical elements (DOEs) for general beam shaping of monochromatic, spatially incoherent light are demonstrated. Direct and indirect methods, i.e., adding a lens' phase to the designed Fraunhofer-type DOEs, are used for the design. The indirect method can reduce the calculation time by approximately half without loss of design accuracy. Two different design examples are shown. For one design the direct method gives a maximum sidelobe intensity of 5.0% of the maximum intensity in the signal window. For the second design the indirect method gives 23.0% of this value. The generated patterns can maintain their basic shapes over a long distance. The elements have been fabricated by directly using gray-scale commercial slides as masks. Experimental results are in close agreement with numerical predictions. PMID- 17086254 TI - Theory and experiments of a tunable wavelength-selective photodetector based on a taper cavity. AB - We demonstrate a wavelength-selective photodetector that combines a Fabry-Perot filtering cavity (FPC) with a taper absorption cavity (TAC). The taper cavity shows a nonresonant effect but exhibits an absorption enhancement effect, so that high speed, high quantum efficiency, wide tuning range, and an ultranarrow spectral linewidth can be achieved simultaneously. Device performance was theoretically investigated by including key factors such as taper angle, finite size diffracting-beam input, and lateral walk-off in the taper cavity. The device was fabricated by bonding a GaAs-based FPC, which can be tuned via thermal-optic effect, with an InP-based TAC. An integrated device with a spectral linewidth of 0.6 nm (FWHM), a wavelength tuning range of 10.2 nm(1518.0-1528.2 nm), a 3 dB bandwidth of 12 GHz, and a quantum efficiency of approximately 70% was demonstrated, and the absorption layer thickness is only 0.3 microm. PMID- 17086255 TI - Non-line-of-sight optical wireless sensor network operating in multiscattering channel. AB - Networks of sensors are envisaged to be major participants in future data gathering systems for civilian and military applications, including medical and environmental monitoring and surveillance, home security, agriculture, and industry. Typically, a very large number of miniature sensing and communicating nodes are distributed ad hoc at the location of interest, where they establish a network and wirelessly communicate sensed data either to one another or to a base station using various network topologies. The optical modality is a potential solution for the links, due to the small and lightweight hardware and low power consumption, as well as other special features. Notably, the backscattering of light by molecules and aerosols in the atmosphere can function as a vehicle of communication in a way similar to the deployment of numerous tiny reflecting mirrors. The scattering of light at solar-blind ultraviolet wavelengths is of particular interest since scattering by atmospheric particles is significant and ambient solar interference is minimal. In this paper we derive a mathematical model of a simple and low-cost non-line-of-sight (NLOS) optical wireless sensor network operating in the solar-blind ultraviolet spectral range. The viability and limitations of the internode link are evaluated and found to facilitate miniature operational sensor networks. PMID- 17086256 TI - Analysis and design of terahertz photonic crystal fibers by an effective-index method. AB - An effective-index method (EIM) is used to analyze and design photonic crystal fibers (PCFs) for the terahertz radiation. By building an analogy between a conventional optical fiber and a PCF, the EIM solves the effective index of the fiber cladding and the effective modal index of a PCF analytically. The EIM is first validated by comparison with available data in the reference, showing that the role of material dispersion is negligible at higher frequencies. Terahertz PCFs with flattened dispersion are designed based on this method and the scaling property of the Maxwell equations. PMID- 17086257 TI - Preconditioned superprism-based photonic crystal demultiplexers: analysis and design. AB - We present the analysis and design of a new type of photonic crystal (PC) demultiplexers (i.e., preconditioned demultiplexer), in which the simultaneous existence of the superprism effect and the negative effective index for diffraction results in a compact structure by canceling the second-order spectral phase to avoid beam broadening inside the PC. This approach considerably relaxes the requirements for the large area of the structure and the small divergence of the input beam. As a result, the size of the preconditioned demultiplexers varies as N(2.5) (N being the number of wavelength channels) compared to the N(4) variation in the conventional superprism-based PC demultiplexers. We use a generalized effective index model to analyze, design, and optimize these demultiplexing structures. This approximate model can be used to extract all the basic properties of the PC device simply from the band structure and eliminates the need to go through tedious simulations especially for three-dimensional structures. Our results show that the preconditioned superprism-based PC demultiplexers have 2 orders of magnitude smaller size compared to the conventional ones. PMID- 17086258 TI - Timing jitter reduction and single-frequency operation in an acousto-optic Q switched Cr,Nd:YAG laser. AB - A laser-diode-pumped watt-level single-frequency combined actively and passively Q-switched (CAPQ) laser with a wavelength of 1064 nm that simultaneously uses a codoped Cr(4+),Nd(3+):YAG crystal as the gain medium and mode selector is presented. The optimum repetition period of the CAPQ laser is approximately 1.5 times the natural repetition period of the passively Q-switched laser. The averaged timing jitter is approximately 400 ns in the CAPQ laser and the repetition rate of the CAPQ laser is experimentally shown to reach a time instability of 0.2% over 10(6) pulses. The output is a stable single frequency with a linewidth of 400 MHz at 20 W pump power. PMID- 17086259 TI - Nano-polymer-dispersed liquid crystal as phase modulator for a tunable vertical cavity surface-emitting laser at 1.55 mum. AB - We demonstrate what we believe is the first nonmechanical tunable vertical-cavity surface-emitting laser operating in the C band. This was achieved as a result of the combination of an InGaAs quantum well structure with a 6lambda thickness tunable index nano-polymer-dispersed liquid-crystal material. Experimental results exhibited a potential tunable range close to 10 nm, in the preliminary version, and excellent single mode locking due to the side-mode suppression ratio (more than 20 dB) over the whole spectral range. Another decisive advantage, compared to mechanical solutions, was the tuning response time of a few tens of microseconds (>30 micros) to scan the full spectral range (10 nm), making this device appropriate for some access network functions, as well as being robust and low cost. The voltage values are the main limitation to wavelength range extension. We present a first version of the device optically pumped. The next version will be electrically pumped as required for the access network applications targeted here. PMID- 17086260 TI - High-bandwidth laser frequency stabilization to a fiber-optic delay line. AB - Stabilization of laser frequency to interferometers with a large time delay in one arm is of significant interest to space-based gravitational wave detectors such as the Laser Interferometer Space Antenna. A recently proposed technique allows a control bandwidth larger than the inverse delay time to be achieved. We present experimental results demonstrating laser frequency stabilization to an optical fiber delay line. A control bandwidth approximately 50 times the inverse delay time is demonstrated. PMID- 17086261 TI - High-frequency intensity modulation in orthogonal polarized dual frequency lasers with optical feedback. AB - High-frequency modulation of laser output intensity is studied with asymmetric feedback induced by the misalignment of an external feedback reflector in an orthogonal polarized dual frequency laser. The fringe frequency of the optical feedback system is seven times higher than that of a conventional optical feedback system, due to multiple feedback effects. The output characteristics of two orthogonal polarized modes are also investigated. Mode competition is observed between the two modes. When initial intensities of the two modes are unequal, the mode competition will be strong. The difference in initial intensity between the two orthogonally polarized modes plays an important role in the mode competition with optical feedback. Experimental results are presented, as well as a theoretical explanation. The high-frequency modulation of laser intensity can greatly increase the resolution of an optical feedback sensing system. PMID- 17086262 TI - SOCRATE: an optical bench dedicated to the understanding and improvement of a laser conditioning process. AB - We present an automatic excimer laser bench (SOCRATE) allowing for the treatment of optical components by laser conditioning. This apparatus, developed at the Commissariat a l'Energie Atomique-Le Ripault, has been designed to add to this conditioning process an in situ, accurate laser-induced damage threshold (LIDT) measurement and different nondestructive optical techniques for the characterization of the component during treatment. Through different examples, we demonstrate the importance of these characterizations to improve the understanding of the laser conditioning. The role of an in situ adapted metrology associated in real time with a laser conditioning bench offers new opportunities to analyze laser-induced damage mechanisms and subsequently to increase the LIDT of optical components. PMID- 17086263 TI - Intracavity generation of radially polarized CO2 laser beams based on a simple binary dielectric diffraction grating. AB - We present experimental results on the intracavity generation of radially polarized light by incorporation of a polarization-selective mirror in a CO(2) laser resonator. The selectivity is achieved with a simple binary dielectric diffraction grating etched in the backsurface of the mirror substrate. Very high polarization selectivity was achieved, and good agreement of simulation and experimental results is shown. The overall radial polarization purity of the generated laser beam was found to be higher than 90%. PMID- 17086264 TI - Characterization of surface-plasmon modes in metal-clad optical waveguides. AB - Finite-element analysis, based on the vector H-field formulation and incorporating the perturbation technique, is used to calculate the complex propagation characteristics of metal-coated dielectric waveguides. The propagation and attenuation characteristics of the surface-plasmon modes at the metal/dielectric interfaces are presented. The effects on the optical properties of metal-clad optical fibers with infinite and finite cladding thickness and the formation of the supermodes due to the coupling between the surface-plasmon modes in the presence of different surrounding materials are also investigated. PMID- 17086265 TI - Fluorescence preselection of bioaerosol for single-particle mass spectrometry. AB - We have designed, constructed, and tested a system that pre-selects the biological fraction of airborne particles from the overall aerosol. The preselection is based on fluorescence emission excited by a continuous 266 nm laser beam. This beam is one of two cw beams used to measure the aerodynamic particle size of sampled particles. The intention in our system is that single particles, based on size and fluorescence emission, can be selected and further examined for chemical composition by mass spectrometry. PMID- 17086266 TI - Multiline absorption spectroscopy for methane gas detection. AB - A multiline absorption spectroscopy technique was investigated based on the single-line absorption spectroscopy technique. An open-path methane-detecting system was designed. An LED was used as a broadband source, and a Fabry-Perot interferometer whose transmission peaks matched the methane R-branch absorption lines was used to enhance the detectable sensitivity. We demonstrate a minimum detectable concentration of 7600 +/- 10% ppm (parts per million) with a multiline differential absorption spectroscopy technique and a concentration of 1000 +/- 10% ppm with a multiline wavelength modulation spectroscopy technique. PMID- 17086267 TI - Diffraction characteristics of a Fresnel zone plate illuminated by 10 fs laser pulses. AB - Results of experimental and theoretical work performed to compare diffraction patterns and focal distributions of a Fresnel zone plate illuminated by ultrashort 10 fs pulsed and cw laser beams are presented. It is shown that the foci intensities of 10 fs pulses are considerably lower than those of cw beams while the focal widths in the axial and radial directions are broadened. Calculations also indicate the spectral modulation along the center of the diffraction patterns. These phenomena are explained by the coherent superposition of the composing frequency content. PMID- 17086268 TI - Tomographic imaging of oxygen by phosphorescence lifetime. AB - Imaging of oxygen in tissue in three dimensions can be accomplished by using the phosphorescence quenching method in combination with diffuse optical tomography. We experimentally demonstrate the feasibility of tomographic imaging of oxygen by phosphorescence lifetime. Hypoxic phantoms were immersed in a cylinder with scattering solution equilibrated with air. The phantoms and the medium inside the cylinder contained near-infrared phosphorescent probe(s). Phosphorescence at multiple boundary sites was registered in the time domain at different delays (t(d)) following the excitation pulse. The duration of the excitation pulse (t(p)) was regulated to optimize the contrast in the images. The reconstructed integral intensity images, corresponding to delays t(d), were fitted exponentially to give the phosphorescence lifetime image, which was converted into the three-dimensional image of oxygen concentrations in the volume. The time independent diffusion equation and the finite element method were used to model the light transport in the medium. The inverse problem was solved by the recursive maximum entropy method. We provide what we believe to be the first example of oxygen imaging in three dimensions using long-lived phosphorescent probes and establish the potential of these probes for diffuse optical tomography. PMID- 17086269 TI - Position of the prism in a dispersion-compensated acousto-optic deflector for multiphoton imaging. AB - The performance of a dispersion-compensated acousto-optic deflector (AOD) for steering femtosecond laser pulses was examined with the prism located before or after the AOD, which is regarded as prism-AOD and AOD-prism, respectively. Comparisons are made over parameters including the spot spatial pattern, output pulse width, scanning linearity, the field of view, and the transmission rate. Fluorescence images of 170 nm diameter beads and cells were measured to provide an overall evaluation for these femtosecond laser beam scanning configurations. On the basis of these experiments, the prism-AOD configuration is concluded to be more advantageous for the purpose of simultaneous compensation for the spatial and temporal dispersion. PMID- 17086271 TI - [Molecular characterization of full-length genome of Japanese encephalitis virus (02-76) newly isolated in China.]. AB - BACKGROUND: To sequence and analyze the complete nucleotide sequence of the Japanese encephalitis virus (JEV) strain 02-76, newly isolated in 2002 in China and to provide information for the genomic structure of JEV and the characteristics of virulence. METHODS: Overlapping primers were designed according to the full-length genomes from GenBank. RT-PCR was used to amplify the fragments, sequencing was performed and all the nucleotides were connected to acquire the full-length genome. Computer software was used to analyze the nucleic acid data, deduced amino acid sequence and phylogenetic trees including Clustal X(1.8), DNASTAR, GENEDOC(3.2). RESULTS: The result of sequence analysis showed that the genome of 02-76 strain was 10,977 nucleotides long. An open reading frame from 95 to 10,391 including 10,296 bases was found capable of coding for a 3432 amino acid polyprotein. Compared with the Beijing 1 strains isolated in 1949 in China, there was a 248 nucleotide divergence and 16 amino acid divergence. Comparison of the complete genome sequences of different JEV isolates showed a 0.6%-15.1% nucleotide sequence divergence among them, which resulted in 0.2%-4.6% amino acid sequence divergence. Phylogenetic analysis through PrM/C,E,3'NTR and full-length genome showed that the 02-76 strain belonged to genotype 3. CONCLUSION: Analysis based on the complete genome sequences of different JEV isolates showed that the 02-76 isolate in 2002 belonged to genotype 3 and was close to the old Chinese isolates SA-14. PMID- 17086272 TI - [Preparation and characterization of the monoclonal antibodies against Japanese encephalitis virus.]. AB - BACKGROUND: To prepare mouse monoclonal antibodies (McAbs) against Japanese encephalitis virus (JEV)and evaluate their biological characteristics. METHODS: McAbs against JEV were prepared by immunizing, fusing, cloning and screening. Their sensitivity, specificity, universality and neutralizing function were analyzed with ELISA, IFA, NT and Western blot. RESULTS: Titers of three McAbs against JEV were higher than 106. Three McAbs only reacted with JEV and not with other nine arboviruses. F12.37 could react with ten strains of JEV and sensitively detected ten replicating strains of viruses in BHK cell. The strains P3 and SH03-103 of JEV were neutralized by F12.37, its titers of protecting 50% cell were 3.2x105 and 105. Western blot showed that F12.37 reacted with envelop(E)protein of JEV. CONCLUSION: Three McAbs against JEV had high titer and good specificity. And F12.37 was very sensitive and universal in reacting with JEV, and neutralized JEV of Genotype I and Genotype .The binding site of F12.37 lays in E protein of JEV. PMID- 17086273 TI - [Preparation of monoclonal antibodies against West Nile virus envelope protein domain.]. AB - BACKGROUND: To prepare monoclonal antibodies against West Nile virus (WNV) envelope protein domain. METHODS: BALB/c mice were immunized with recombinant antigen of West Nile virus envelope protein domain, and the spleen cells of the mice were used to prepare the monoclonal antibodies (McAb) by hybridoma technique. RESULTS: Three hybridoma cell strains secreting McAbs against WNV envelope protein domain, designated as 4F7, 6H3 and 8E4, respectively, were obtained and were identified by indirect enzyme linked immunosorbent assay (ELISA), they belonged to IgG1, IgG1 and Ig2a, respectively. Two epitopes of envelope protein domain were determined, among them, 4F7 and 6H3 were against the same epitope and 8E4 to another one. CONCLUSIONS: The results of indirect ELISA, Western blot and indirect immunofluorescence experiment indicated that these three McAbs were specific for West Nile virus envelope protein domain and did not cross-react with Japanese encephalitis virus and other viruses, so they can be used for specific detection of West Nile virus. PMID- 17086274 TI - [Epidemic situation analysis of Japanese encephalitis in 2005.]. AB - BACKGROUND: To investigate the epidemic situation of Japanese encephalitis (JE) in three provinces, Guizhou, Sichuan, and Hubei in 2005. METHODS: Information about epidemic situation of JE, mosquitoes specimens were collected and titers of JE virus in hosts in the above three surveillance sites were determined. RESULTS: The reported cases of JE in Guizhou, Sichuan, and Hubei province accounted for 40.7% of total cases in 2005 in China. The numbers of cases in Guizhou and Sichuan ranked at the first two in China, morbidity exceeded 1/100,000, which was higher than average level in China. Zero to 10 years old children accounted for 90% in reported cases. Most of the cases were children lived at home. Almost all JE cases were presented from June to September, but most cases were reported between July and August. Investigaton of the density of vector showed that the dominant mosquitoes were Culex, especially the Culex tritaeniorhynchus. CONCLUSION: The epidemic status of JE was similar among the three provinces and the whole country. The number of JEV cases in Guizhou and Sichuan were the highest in China. PMID- 17086275 TI - [Effects of antiviral effects of agents and prognosis of chronic hepatitis B.]. AB - BACKGROUND: To analyze the effects of difference antiviral agents and the effects of the treatments on long-term prognosis. METHODS: Retrospective research method was applied. RESULTS: About 40% of the patients were treated with interferon or lamivudine. After the treatment, in lamivudine group, the negative rate of HBV DNA was the highest. In the interferon group, the sero conversion rates of HBeAg/HBeAb were 22.9%. In the antiviral treatment patients, the disease progression and the occurrence of cirrhosis and liver cancer were much lower than those of the control groups. The mortality of cirrhosis and liver cancer in the HBeAg/HBeAb sero converted group was much lower than that of the group without HBeAg/HBeAb sero conversion groups (P less than 0.05). CONCLUSION: The antiviral effects of interferon and lamivudine were better than those of the other drug groups. The antiviral drugs could relieve the disease progression and reduce the mortality of cirrhosis and liver cancer. PMID- 17086276 TI - [Subtype and sequence analysis of the ENV gene of HIV-1 drug users in Guangdong Pearl River delta.]. AB - BACKGROUND: To determine the subtype of HIV-1 of drug users (DUS) in Guangdong Pearl River delta. METHODS: HIV-1 pro-viral DNA from buffy coat of 43 DUS in Guangdong was amplified by nested PCR. The C2-V3 regions of HIV-1 ENV gene was sequenced directly from the PCR product and analyzed. RESULTS: The 43 DUS were confirmed to be infected with four HIV-1 subtype or Circulating Recombinant Form (CRFs): 07-BC(n=29), AE(n=9), 08-BC(n=3) and B(n=2). Genetic distances showed that the AE group was the closest to CM240 strain isolated in Thailand which is mainly circulating in sexually transmitted infector. The 07-BC group was the closest to C54A strain isolated in Northeastern China. The 08-BC group was the closest to 97CNGX-9F strain isolated in Guangxi, China. The B strain was the closest to rl42 strain isolated in Thailand. CONCLUSION: HIV-1 CRFs 07-BC predominates in DUS in Guangdong Pearl River delta. PMID- 17086277 TI - [Research on development of gold immunochromagraphic assay test kit for serum Coxsackievirus IgM antibody.]. AB - BACKGROUND: To develop gold immunochromatographic assay (GICA) test kit for serum Coxsackievirus IgM antibody and optimize the key experiment conditions. METHODS: Colloidal gold particles of 20 nm were prepared and coupled with sheep anti-human IgM, the gold immunochromatography assay test kit was setup for serum Coxsackievirus IgM antibody. RESULTS: The spheroidal colloid gold particles of 20 nm showed bright red in color. The minimal stable concentration (MSC) of gold sheep anti-human IgM was 1 ug/ml, and the suitable stable concentration (SSC) was 1.5 ug/ml. The pH 8.2 was appropriate; 30 sera samples were tested by GICA and ELISA, there was no significant difference between the two methods. CONCLUSION: The quality of gold immunochromatography assay test kit is associated with such factors as the quality and quantity of antigen or antibody, colloid gold particles, buffers, etc. PMID- 17086278 TI - [Relationship between hepatitis B virus genotypes and basic core promoter/precore mutations in patients with severe hepatitis.]. AB - BACKGROUND: To investigate the relationship between hepatitis B virus genotypes and basic core promoter (BCP)/precore mutations in patients with severe hepatitis. METHODS: HBV genotypes were determined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) in 52 patients with severe hepatitis and 52 cases with chronic hepatitis B (CHB). Eight samples of genotypes B and C were randomly selected and their S gene was directly sequenced, and then their phylogenetic trees were analyzed. Fifteen samples of each of genotypes B and C were randomly selected and their BCP and precore genes were directly sequenced with PCR, and then the relationship between genotypes and BCP(T1762/A1764)/precore (A1896) mutations were analyzed. RESULTS: Genotype B C and mixed genotypes (B and C) were detected in patients with severe hepatitis in Quanzhou area. Genotype B was the majority with a proportion 48.08% others had a proportion 30.77% and 17.31%, respectively. Genotype A, E and F were not detected. The percents of genotype C, mixed B and C in severe hepatitis were significantly higher than that in CHB. The double mutation in BCP (T1762/A1764) was significantly more frequent in severe hepatitis with genotype C than that in genotype B (P less than 0.05). However, there was no significant difference in the distribution of precore mutant with A1896 between genotype B and C patients (P greater than 0.05). CONCLUSION: Genotype C may induce more severe liver inflammation than that genotype B may do. Mixed genotypes B and C infection may be an important determinant of inducing severe hepatitis. The double mutation in BCP (T1762/A1764) was more common in severe hepatitis with genotype C than that with genotype B. PMID- 17086279 TI - [Establishment of Z-HL16C cell line.]. AB - BACKGROUND: To establish and study the nature and the application of Z-HL16C cell line. METHODS: The cell line was continuously passed, frozen stored and recovered. Its application was expanded and the cell type was identified. RESULTS: The cell line had an epithelial-cell-like shape, the size appeared uniform, the cell boundary was distinct. It has been continuously passed, frozen stored and recovered for ten years. Its recovery rate was about 90%. It has been proved to be sensitive to the tested viruses which were enteroviruses (Polio, Cox, Echo), influenza viruses, parainfluenzaviruses, adenoviruses, measles virus. This cell line has been identified as a cancerization cell. CONCLUSION: The cell line Z-HL16C has been stably established, it has a broad spectrum in sensitivity for culturing viruses. PMID- 17086280 TI - [Detection of serum hepatitis B virus large envelope protein and its relationship with viral replication.]. AB - BACKGROUND: To explore the significance of HBV large envelope protein (LHBs) in diagnosing HBV replication in chronic hepatitis B patients. METHODS: Serum HBV DNA was quantitively detected by using real-time polymerase chain reaction (RT PCR), the LHBs and Pre-S1 were detected by using enzyme linked immunosorbent assay (ELISA) and HBV markers were detected by time differentiate immunofluorescence assay in 340 serum samples collected from chronic hepatitis B patients. RESULTS: Serum LHBs level was closely correlated with number of HBV DNA copies (r=0.899, P=0.0380). There was no significant difference between positive rate of LHBs and that of HBV DNA in different HBeAg pattern (P>0.05); the positive rate of LHBs was 83.15%, which was higher than that of Pre-S1 and HBeAg which were 50.54% and 54.48%, respectively. There was significant difference (P less than 0.05). CONCLUSION: The level of serum LHBs can be used to estimate the state of HBV replication and the sensitivity was superior to both Pre-S1 and HBeAg. So it may be used as a new serological marker to detect HBV replication. PMID- 17086281 TI - [Detection of plasma urokinase-type plasminogen activator and its receptor in acute or chronic hepatitis B patients and its clinical significance.]. AB - BACKGROUND: To study the expression of plasma uPA and uPAR in Acute or Chronic Hepatitis B patients, to investigate changes of blood fibrinolytic system in Hepatitis B and its clinical significance. METHODS: The level of detected plasma urokinase-type plasminogen activator (uPA) and urokinase-type plasminogen activator receptor (uPAR) were measured by Enzyme Linked Immuno Sorbent Assay (ELISA). RESULTS: The levels of plasma uPA and uPAR were significantly higher in patients with Acute or Chronic Hepatitis B than that of control group (P less than 0.05, P less than 0.01), and also higher in serious group than that of non serious group of Chronic Hepatitis B patients (P less than 0.05), and also higher in chronic group than in acute group (P less than 0.05). The level of plasma uPAR was significantly higher in patients with serious jaundice than that of non serious jaundice group (P less than 0.01). The levels of uPA and uPAR are significantly higher in acute period than that in recover period of Acute Hepatitis B patients, but the levels in recover period is also higher than that in control group. The level of plasma uPAR have positive correlation with prothrombin (PT), international normalized ratio (INR) and total bilirubin (P less than 0.01). CONCLUSION: The level of plasma UPA nad UPAR were enhanced in Acute or Chronic Hepatitis B patients was related with the degree of inflammation, was also related with high expression of serious damage of liver cell. It may be one of important cause to imbalance between coagulation and fibrinolytic in hepatitis. PMID- 17086282 TI - [Distribution of hepatitis B virus genotypes in Guizhou and analysis of clinical significance.]. AB - BACKGROUND: To investigate the distribution of HBV genotypes in Guizhou and study the relationship between the genotype and the progression of liver disease. METHOD: Totally 693 patients with chronic HBV infection, from 4 areas of Guizhou, including 292 asymptomatic HBV carriers (ASC), 276 cases with chronic hepatitis (CH), 76 liver cirrhosis (LC), 49 hepatocellular carcinoma (HCC) were examined. The HBV genotype was determined by restriction fragment length polymorphism analysis and the relationship between HBV genotype and the progression of liver disease was studied by multifactor analysis such as HBsAg positivity, HBV DNA load and ALT level. RESULTS: Of the 693 patients, 6 (0.87%), 449 (64.79%), 233 (33.62%), and 5 (0.72%) belonged to genotype A, B, C and D, respectively. There was a statistically siganificant difference in the distribution of genotype B among Kaili (96.4%), Zunye (78.79%), Duyun (76.19%) and Guiyang (53.66%) areas (P less than 0.01). Genotype C was more prevalent in Guiyang than in other three areas (45.68% vs. 23.8%, 45.68% vs. 13.13%, 45.68% vs. 3.96%, P less than 0.01, respectively). There were also statistically significant difference in distribution of genotype B and C in various stages of liver disease (P less than 0.01). Genotype B showed a decreasing trend from ASC, CH, LC to HCC and genotype C was more frequent in patients with CH (45.92%) than in those with ASC (20.6%), LC (17.17%) and HCC (16.31%). The ALT levels and the mean ages were significantly higher and older in patients with genotype C than those in genotype B. The HBeAg positivity was significantly lower in genotype C than that in genotype B (48.07% vs 61.90%, P less than 0.01). CONCLUSION: Genotype A, B, C and D exist in Guizhou. Genotype B predominates and genotype C was the second common. The geographic distribution of genotype B and C are different in some areas of Guizhou. Compared with genotype B, genotype C is associated with the development of more severe liver damage. PMID- 17086283 TI - [Comparative study on clinical features of 215 patients with acute and subacute severe hepatitis.]. AB - BACKGROUND: To analyze the clinical features of acute severe hepatitis (ASH) and subacute severe hepatitis (SSH) by comparison analysis. METHODS: The clinical features of 215 cases with ASH and SSH from 1995 to 2005 were retrospectively analyzed by using chi-test, t-test and regression analysis. RESULTS: The most common cause of ASH or SSH was hepatitis B virus (HBV) infection (>30%). Anti tuberculosis therapy was the main cause of drug-induced ASH or SSH. The incidences of hepatic encephalopathy were 78.13% and 43.05% respectively (P less than 0.001). Prothrombin activity was less than 20% and less than 30%, respectively in cases with ASH and SSH. The most common complications of ASH were hepatic encephalopathy, imbalance of electrolyte and cerebral edema, and that of SSH were ascites, imbalance of electrolyte and hepatic encephalopathy. Prothrombin time, ratio of WBC and neutrophil count were positively related to case fatality of ASH or SSH. Prothrombin activity and total cholesterol (TC) were negatively correlated with fatality of ASH or SSH. Total bilirubin, blood ammonia and creatinine were also positively correlated with fatality of SSH; chlolinesterase, triglyceride, PLT, albumin were negatively correlated with fatality of SSH. CONCLUSION: ASH and SSH are two different entities because of obvious differences in the ages of suffering, the incidence and occurring time of hepatic encephalopathy, abnormality of function of blood coagulation and the marker for predicting prognosis. Severe abnormality of function of blood coagulation is an important marker with high sensitivity and specificity in ASH or SSH patients without hepatic encephalopathy. PMID- 17086284 TI - [Detection and analysis of HBV antigen protein in kidney tissue and HBV DNA in serum and kidney tissue of patients with HBsAg+ IgA nephropathy.]. AB - BACKGROUND: To explore the relationship between hepatitis B virus (HBV) infection and development of IgA nephropathy. METHODS: HBsAg and HBcAg protein in renal biopsy specimens of 32 cases was detected on frozen sections and HBV DNA was detected in paraffin section of renal biopsies and in serum of 42 HBsAg positive cases. RESULTS: The positive rate of HBAg in renal biopsies of IgA nephropathy was 59.1%, and 63.6% in non-IgA nephropathy, there was no significant difference between them. In 42 cases biopsies of renal tissues, only five were HBV-DNA positive (11.9%). The five cases were HBsAg, HBcAb and HBeAg positive, the pathological diagnosis of two cases were mesangial proliferative glomerulonephritis; one had minimal change of glomerulonephritis; and one had basement membrane change; and only one had IgA nephropathy. At the same time, in 42 HBsAg+ cases the serum specimens were detected; 12 cases were positive for HBsAg, HBcAg and HBeAg, in whom serum HBV-DNA was positive, but only 5 were positive for HBV-DNA in renal biopsy tissue, and HBV-DNA was negative in other 30 blood serum and tissue specimens. CONCLUSION: The difference in expression of HBsAg, HBcAb and HBeAg protein between IgA nephropathy and non-IgA nephropathy tissue from renal biopsy was not significant. There is no direct relationship between HBV infection and IgA nephropathy. PMID- 17086285 TI - [Phenotype of peripheral blood mononuclear cells derived dendritic cells from patients with chronic hepatitis B.]. AB - BACKGROUND: The aim of this study was to access phenotype changes of dendritic cells (DC) cultured from peripheral blood mononuclear cells (PBMC) in patients with chronic hepatitis B and to reveal the relationship between phenotype of DC and ALT or HBV DNA. METHODS: Indices of ALT and serum HBV DNA were measured in 37 patients with chronic hepatitis B and 21 healthy controls. Peripheral blood mononuclear cells were isolated from all patients and healthy controls, and cultured with granulocyte-macrophage colony-stumilating factor (GM-CSF), interleukin-4 (IL-4) and tumor necrosis factor- (TNF-)in RPMI 1640 medium that contained 10% fetal calf serum. After culturing for 7 days, the DC was counted and the phenotypes were detected by FACS. Then the data were statistically analysed. RESULTS: The DC was significantly fewer (P less than 0.05) in patients with chronic hepatitis B than the controls. In particular, the expressive level of CD83 and CD86 on DC's surface from patients with chronic hepatitis B were also significantly lower (P less than 0.05) than that from the controls. In the patients with hepatitis B, the indices of DC had a significantly negative correlation with the level of serum HBV DNA (P less than 0.05), but no significant relationship was found between ALT and indices of DC (P greater than 0.05). CONCLUSION: The DC cultured from patients with chronic hepatitis B were few and had immature phenotype. These changes had a significantly negative correlation with the level of serum HBV DNA, but had not correlation with the inflammatory reaction levels in the liver. DC was associated with the clearance of HBV in patients with hepatitis B. PMID- 17086286 TI - [Expression of total RNA and amplification of myocardial troponin gene during monitoring viral cardiomyocyte injury.]. AB - BACKGROUND: To investigate the value of circulating cTnI-mRNA detection for monitoring myocardial injury development and prognosis. METHODS: Viral myocardial injury models in BALB/c mice were created by intraperitoneal inoculation with Coxsackievirus B3 (CVB3,1x108 TCID50) for inducing myocardial injury. The total RNAs were extracted and cTnI-mRNA in mice cardiac tissues and circulating blood were amplified by RT-PCR during mice myocardial injury. RESULTS: In virus infected mice, the mRNA abundance for cTnI was up-regulated in heart and circulating blood and associated with salient myocardial histopathologic features, including myocardial swelling, inflammatory cell infiltration, pyknosis, karyorrhexis, karyolysis, denaturalization, necrosis, and calcification. The cTnI-mRNA form infected mice heart and circulating cardiac myocytes were analyzed by RT-PCR, the amplified gene fragments were found in all heart tissues. The incidence of cTnI-mRNA was 0, 0, 0, 16.7%, 40.0%, 71.4%, 83.3% and 87.5% in the controls, the 3rd, 6th, 9th, 12th, 15th, 18th,and 21st day in circulating bloods from the infected mice, respectively. CONCLUSION: The present data suggest that cTnI-mRNA expression is up-regulated and released into blood on viral myocardial injury, and detection of circulating cTnI-mRNA is a sensitive genetic marker for monitoring myocardial injury development and prognosis. PMID- 17086287 TI - [Production and identification of the monoclonal antibodies specific for MHC class I complexes bound with HPV16E7 CTL epitopic peptide.]. AB - BACKGROUND: To produce specific monoclonal-antibodies (MAbs) for MHC class I complexes bound with HPV16E7 CTL epitopic peptide (49-57), and provide a foundation for the investigation of the present pathway of viral antigen protein after in vitro viral infection. METHODS: Highly purified HPV16E7CTL epitope (49 57) (RAHYNIVTF) was produced, and then TAP deficient RMA-S cells incubated with RAHYNIVTF were used to immunize the BALB/c mouse. The spleen cells of the mice were regularly harvested and fused with the SP2/0 cells. The growing fusion wells were screened and the abstracted Mabs were identified in terms of sensitivity, specificity and affinity. RESULTS: The screened hybriroma cells could steadily secrete the MAbs specific for MHC class I complexes bound with HPV16E7CTL epitopic peptide. The MAbs showed high reactivity with TAP-deficient RMA-S cells loaded with RAHYNIVTF and RMA-S cells which have the ability to process the endogenous MHC class I complexes, while minimally bound to class I molecules bearing other peptides, the results indicated excellent sensitivity, specificity and affinity of the MAbs. CONCLUSION: The experiments provide a method for producing MAbs for epitopic peptide bound MHC class I complexes. PMID- 17086288 TI - [Synergistic effects of human papillomavirus and phorbol ester in cell transformation.]. AB - BACKGROUND: To investigate the molecular mechanism of the synergistic effects of human papillomavirus (HPV) and phorbal esters (TPA) in cell transformation. METHODS: The expression of oncogenes and anti-oncogenes in 293 cell line treated with HPV and TPA was studied by Southern Blot and RNA dot blot. RESULTS: It was found that the synergistic effect induced the amplification of c-myc (4-8 times), increased expressing level of c-erbB-2 (32-64 times) and decreased expressing level of p16(1/4-1/8). CONCLUSION: The above results show that the synergistic effect has an important role in development of carcinoma. PMID- 17086289 TI - [Comparison of IgA/VCA, IgA/EA, IgG/EA in immunoenzyme methods and ZEBRA ELISA in early diagnosis of nasopharyngeal carcinoma.]. AB - BACKGROUND: To develop an ELISA method using Herpesvirus hominis type 4 (EBV) IgG/Zebra as capture antigen for large population screening. METHODS: The ELISA method used purified ZEBRA antigen to detect the IgG/ZEBRA antibody from serum in nasopharyngeal carcinoma (NPC) and normal healthy subjects. RESULTS: Of 288 NPC sera, 262 were detected positive, the sensitivity was 91%, while 5 of 96 normal sera were detected positive, the specificity was 94.8% and the results of NPC group and healthy group displayed significant difference (P less than 0.001). IgA/VCA, IgA/EA, IgG/EA in immunoenzyme methods and ZEBRA ELISA were compared during the NPC screening in two cities: Huizhou, Guangdong and Guiping, Guangxi, 5463 and 2017 samples respectively were tested and 5 earlier NPC patients were found. CONCLUSION: The results indicate that this method has high specificity and sensitivity, and can be used for large population screening to assist early phase NPC diagnosis. PMID- 17086290 TI - [Codon usage of Chinese hamster ovary cells.]. AB - BACKGROUND: To investigate the optimal codons of dihydrofolate reductase (dhfr) minus Chinese hamster ovary cells (CHO dhfr-). METHODS: A cDNA library of CHO dhfr- containing high abundence mRNA was constructed and protein-coding sequences were obtained after identification and analysis. Codon frequence of CHO dhf- was compared with that of Chinese hamster in CUTG database. Then codon usage variation among cDNA was investigated using correspondence analysis (COA). RESULTS: Fifty qualified cDNAs from CHO dhfr- were selected, which encodes proteins of high abundence. Comparing with the codon frequence of Chinese hamster, the highest frequence of synonymous codons for amino acids in CHO dhfr- cells were the same as Chinese hamster except that of Arg and Pro. This method of COA identifies the first main factor which can account for the largest fractions (14.7%) of variation among cDNAs. Twenty-two synonymous codons were identified as the optimal codons of CHO cell. CONCLUSION: CHO dhfr- cell has its own optimal codons, it is suggested that codon bias is one of reasons for functional diversity of different mammal cells and it is an effective stratagy to modification of the codon usage of the foreign gene according to the optimal codons of CHO dhfr- to increase the production of foreign gene. PMID- 17086291 TI - [Cloning of genes transactivated by nonstructural protein 4A of hepatitis C virus.]. AB - BACKGROUND: To construct a subtractive cDNA library of genes transactivated by NS4A protein of hepatitis C virus with suppression subtractive hybridization technique (SSH). METHODS: The mRNA was isolated from Hep G2 cells transfected pcDNA3.1(-)-NS4A and pcDNA3.1(-) empty vector, respectively, then the cDNA was synthesized. SSH method was employed to analyze the differentially expressed RNA sequence between the two groups. The twice enriched PCR products were subcloned into T/A vectors to set up the subtractive library. Amplification of the library was carried out with E.coli strain JM109. The cDNA was sequenced and analyzed in comparison with those in GenBank with Blast search after PCR. RESULTS: The amplified library contained 36 positive clones. Colony PCR showed that 36 clones contained 200-1000 bp inserts. Sequence analysis was performed in 25 clones, and the full length sequences were obtained with bioinformatics method. Altogether 20 kinds of coding sequences were achieved, which consisted of 18 kinds of known and 2 kinds of unknown ones. The obtained sequences may be target genes transactivated by NS4A protein of HCV, among which some genes coding for proteins involved in cell cycle regulation, cell apoptosis, signal transduction pathway and tumour development. CONCLUSION: A subtractive library of genes transactivated by NS4A protein of HCV was constructed successfully, which brought some new clues for studying the biological functions and pathogenesis of the viral proteins. PMID- 17086292 TI - [Screening and cloning of the down-regulation gene by recombinant interferon-B using suppression subtractive hybridization technique.]. AB - BACKGROUND: To construct a subtractive cDNA library of target genes down regulated in human hepatocarcinoma cell line HepG2 cells treated with IFNB, and clone genes of the down-regulation by IFNB using suppression subtractive hybridization (SSH) technology and bioinformatics techniques. METHODS: The mRNA was isolated from HepG2 cells induced by recombinant interferon-B and 0.9 percent sodium chloride, respectively, then cDNA was synthesized. After restriction enzyme Rsa I digestion, small sizes cDNAs were obtained. Then tester cDNA was divided into two portions and each was ligated to the specific cDNA adaptor 1 and adaptor 2 respectively. After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR, the DNA fragment was subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E.coli strain DH5a. The cDNA was sequenced and analyzed in GenBank with Blast search after PCR. RESULTS: The subtractive library of genes down-regulation in HepG2 cells treated with recombination interferon-B was constructed successfully. The amplified library contained 58 positive clones. Colony PCR and sequence analysis was performed in 35 clones randomly, and the full length sequences were obtained with bioinformatics method. Altogether 12 coding sequences were obtained. CONCLUSION: A subtractive cDNA library of genes down-regulation in HepG2 cells treated with IFNB using SSH technique was constructed successfully, which brings some new clues for studying the regulation mechenism of IFNB in liver cells. PMID- 17086293 TI - [Significance of hepatitis B virus PreS1-Ag, PreS2-Ag, large protein, PreS2-Ab detection and the prediction of HBV DNA replication.]. AB - BACKGROUND: To explore the significance of hepatitis B virus PreS1-Ag, PreS2-Ag, large protein (LP) detection and the prediction of viral replication. METHODS: PreS1-Ag, PreS2-Ag, LP and HBV markers were measured by enzyme linked immunosorbent assay (ELISA) in 201 cases of infected serum. Serum HBV DNA level was quantitatively detected by real-time polymerase chain reaction (PCR). RESULTS: There were significant differences in positive rate between the PreS1 Ag, PreS2-Ag, LP, and HBsAg; the positive rate of PreS2-Ag and LP were higher than that of the HBeAg. No significant differences were found in the positive rates between LP and the levels of HBV DNA and there was a positive correlation between quantitations of HBV DNA and HBV-LP. CONCLUSION: Serum PreS1-Ag, PreS2-Ag and LP were laboratory markers that can accurately reflect HBV DNA reproduction, and were helpful complementarity to traditional HBV M. There is a close correlation between the number of copies of HBV DNA and the levels of HBV-LP. PMID- 17086294 TI - [Clinical evaluation of HIV-1/2 antibody rapid tests.]. AB - BACKGROUND: To evaluate the sensitivity and specificity of HIV-1/2 antibody rapid tests prior to licensing. METHODS: Four hundred forty, 264 and 300 blood samples were collected from normal blood donors, drug addicted individuals and HIV infected patients, respectively. All the samples were tested with 17 kinds of HIV 1/2 antibody rapid tests prior to licensing. The sensitivity and specificity of each test were calculated. RESULTS: The sensitivity for all evaluated rapid tests was 100% for detecting 300 strong positive samples for HIV antibody; the specificity for all evaluated rapid tests was within the range of 97.7%-100% for detecting 440 samples from normal blood donors; the sensitivity and specificity of all tests were within the range of 70.3%-95.1% and 59.1%-100% respectively for detecting 264 samples from HIV high risk population. CONCLUSION: The sensitivity and specificity of different tests were different and the samples from HIV high risk population may be very useful for evaluation of the quality of HIV-1/2 antibody rapid tests. PMID- 17086295 TI - [Development of a rapid subtype-screening assay for the env region of HIV-1 CRF strains in Guangxi.]. AB - BACKGROUND: To develop a simple and rapid subtype-screening assay for the env region of the circulating recombinant form (CRF) of human immunodeficiency virus type 1 (HIV-1) in Guangxi. METHODS: Proviral DNA from HIV-1 positive samples were extracted and subjected to the first round PCR with universal primers for the env region that can detect HIV-1 M group isolates. In the second round PCR, two pairs of subtype-specific primers that were designed to detect subtype C or B'/C and CRF01-AE respectively were added into one tube. The PCR products of different subtypes could be distinguished in agarose-gel electrophoresis. Additionally, all of these samples were sequenced and analyzed phylogenetically. RESULTS: Phylogenetic analysis of the env region of 50 samples showed that 3 samples (6%) were infected with CRF08-BC, 43 (86%) with CRF01-AE, and 4 (8%) remained unclassifiable. Detection of the subtype-specific primer sets revealed that 3 were subtype C or B'/C (100%), 39 were CRF01-AE (90.7%), with an adequate sensitivity (91.3%) and a high specificity (100%). Non-specific bands occasionally appeared but did not interfere with interpretation of the results. The phylogenetic analysis was consistent with subtype-specific primer sets and the consistency rate was 92%. The average reproducibility was 100% for CRF08-BC samples and 93.8% for CRF01-AE samples. CONCLUSION: A simple, rapid and low cost assay was developed for subtype-screening of CRF01-AE in Guangxi. PMID- 17086296 TI - Obesity surgery and satiety control. PMID- 17086297 TI - Gestational prognostic factors in women with recurrent spontaneous abortion. AB - CONTEXT AND OBJECTIVE: Recurrent spontaneous abortion (RSA) is defined as three or more consecutive pregnancy losses before 20 weeks and is associated with several etiological factors related to genetics, anatomy, hormones, infections and immunology, for example. Many cases of RSA remain unclear. New factors or their associations may influence gestational results. The aim was to identify possible single or associated causes of RSA that could predict gestational prognosis for women undergoing investigation and treatment. DESIGN AND SETTING: Case-control study, at the Recurrent Abortion Outpatient Clinic, Department of Obstetrics and Gynecology School of Medicine, Universidade Estadual de Campinas (Unicamp). METHODS: Two hundred and forty-six medical records of women with RSA seen at the Recurrent Abortion Outpatient Clinic, Department of Obstetrics and Gynecology School of Medicine, Universidade Estadual de Campinas (Unicamp), between 1994 and 2003, were evaluated. Data on age, obstetric history, possible etiological factors, treatment and pregnancy outcomes were evaluated. Statistical analysis was performed using odds ratios (OR), logistic regression analysis and decision trees. RESULTS: Two hundred and twenty-nine women were included in the study. The most frequently found etiological factors were immunological, particularly alloimmune factors (93.9%). Women with a single alloimmune factor had better gestational results (77.7% deliveries) than those with other associated factors. Autoimmune factors were associated with a higher abortion rate (OR: 4.30; 95% confidence interval, CI: 1.36-13.63). No association was found between the number of abortions prior to treatment and pregnancy results. Women aged 40 or over presented the highest rate of spontaneous abortion (OR: 5.83; 95% CI: 1.12-30.40). CONCLUSION: Age over 40 years old, immunological factors and two or more concomitant factors were associated with poor gestational outcomes among the women studied. PMID- 17086298 TI - Impact on hospital mortality and morbidity of right ventricular involvement among patients with acute left ventricular infarction. AB - CONTEXT AND OBJECTIVE: There is uncertainty regarding the risk of major complications in patients with left ventricular (LV) infarction complicated by right ventricular (RV) involvement. The aim of this study was to evaluate the impact on hospital mortality and morbidity of right ventricular involvement among patients with acute left ventricular myocardial infarction. DESIGN AND SETTING: Prospective cohort study, at Emergency Care Unit of Hospital Central da Irmandade da Santa Casa de Misericordia de Sao Paulo. METHODS: 183 patients with acute myocardial infarction participated in this study: 145 with LV infarction alone and 38 with both LV and RV infarction. The presence of complications and hospital death were compared between groups. RESULTS: 21% of the patients studied had LV + RV infarction. In this group, involvement of the dorsal and/or inferior wall was predominant on electrocardiogram (p < 0.0001). The frequencies of Killip class IV upon admission and 24 hours later were greater in the LV + RV group, along with electrical and hemodynamic complications, among others, and death. The probability of complications among the LV + RV patients was 9.7 times greater (odds ratio, OR = 9.7468; 95% confidence interval, CI: 2.8673 to 33.1325; p < 0.0001) and probability of death was 5.1 times greater (OR = 5.13; 95% CI: 2.2795 to 11.5510; p = 0.0001), in relation to patients with LV infarction alone. CONCLUSIONS: Patients with LV infarction with RV involvement present increased risk of early morbidity and mortality. PMID- 17086299 TI - Preliminary results from digestive adaptation: a new surgical proposal for treating obesity, based on physiology and evolution. AB - CONTEXT AND OBJECTIVE: Most bariatric surgical techniques include essentially non physiological features like narrowing anastomoses or bands, or digestive segment exclusion, especially the duodenum. This potentially causes symptoms or complications. The aim here was to report on the preliminary results from a new surgical technique for treating morbid obesity that takes a physiological and evolutionary approach. DESIGN AND SETTING: Case series description, in Hospital Israelita Albert Einstein and Hospital da Policia Militar, Sao Paulo, and Hospital Vicentino, Ponta Grossa, Parana. METHODS: The technique included vertical (sleeve) gastrectomy, omentectomy and enterectomy that retained three meters of small bowel (initial jejunum and most of the ileum), i.e. the lower limit for normal adults. The operations on 100 patients are described. RESULTS: The mean follow-up was nine months (range: one to 29 months). The mean reductions in body mass index were 4.3, 6.1, 8.1, 10.1 and 10.7 kg/m2, respectively at 1, 2, 4, 6 and 12 months. All patients reported early satiety. There was major improvement in comorbidities, especially diabetes. Operative complications occurred in 7% of patients, all of them resolved without sequelae. There was no mortality. CONCLUSIONS: This procedure creates a proportionally reduced gastrointestinal tract, leaving its basic functions unharmed and producing adaptation of the gastric chamber size to hypercaloric diet. It removes the sources of ghrelin, plasminogen activator inhibitor-1 (PAI-1) and resistin production and leads more nutrients to the distal bowel, with desirable metabolic consequences. Patients do not need nutritional support or drug medication. The procedure is straightforward and safe. PMID- 17086300 TI - Complications and risk factors in transrectal ultrasound-guided prostate biopsies. AB - CONTEXT AND OBJECTIVE: Prostate biopsy is not a procedure without risk. There is concern about major complications and which antibiotics are best for routine use before these biopsies. The objective was to determine the rate of complications and the possible risk factors in prostate biopsies. DESIGN AND SETTING: Prospective study, Faculdade de Medicina de Botucatu. METHODS: Transrectal ultrasound (TRUS) guided prostate biopsies were carried out in 174 patients presenting either abnormality in digital rectal examinations (DRE) or levels higher than 4 ng/ml in prostate-specific antigen (PSA) tests, or both. RESULTS: Hemorrhagic complications were the most common (75.3%), while infectious complications occurred in 19% of the cases. Hematuria was the most frequent type (56%). Urinary tract infection (UTI) occurred in 16 patients (9.2%). Sepsis was observed in three patients (1.7%). The presence of an indwelling catheter was a risk factor for infectious complications (p < 0.05). Higher numbers of biopsies correlated with hematuria, rectal bleeding and infectious complications (p < 0.05). The other conditions investigated did not correlate with post-biopsy complications. CONCLUSIONS: Post-biopsy complications were mostly self-limiting. The rate of major complications was low, thus showing that TRUS guided prostate biopsy was safe and effective. Higher numbers of fragments taken in biopsies correlated with hematuria, rectal bleeding and infectious complications. An indwelling catheter represented a risk factor for infectious complications. The use of aspirin was not an absolute contraindication for TRUS. PMID- 17086301 TI - Metastatic breast cancer: do current treatments improve quality of life? A prospective study. AB - CONTEXT AND OBJECTIVE: In metastatic breast cancer cases, the currently available therapeutic approaches provide minimal improvement in survival. As such, quality of life (QOL) becomes one of the main objectives of treatment. It is not known whether current treatments derived from trials improve QOL. The aim was to evaluate changes in QOL among metastatic breast cancer patients receiving treatment derived from trials. DESIGN AND SETTING: Prospective observational QOL survey in a tertiary cancer center. METHODS: To evaluate the influence of current treatments on patients' QOL, the Medical Outcomes Study Short Form-36 (SF-36) and the Beck Depression Inventory (BDI) were applied on three occasions: before starting treatment and at the 6th and 12th weeks, to consecutive metastatic breast cancer patients over a one-year period. RESULTS: We found an improvement in QOL in the sample evaluated (n = 40), expressed by changes in the overall SF 36 score (p = 0.002) and the BDI (p = 0.004). Taken individually, the SF-36 components Pain, Social Functioning and Mental Health also improved significantly. Patients with worse initial performance status and secondary symptoms displayed greater improvement than those with better initial performance status and asymptomatic disease (p < 0.001). Patients who received more than one type of therapy showed larger gains than those given only one type (p = 0.038). CONCLUSIONS: In our environment, current metastatic breast cancer treatments can improve QOL, especially among symptomatic patients and those with low performance status. PMID- 17086302 TI - The burden of injury in Brazil, 2003. AB - CONTEXT AND OBJECTIVE: Since 1980, injuries have been the second biggest cause of death among the Brazilian population. This study aimed to analyze national data on fatal injuries and nonfatal injury hospitalization in Brazil, for 2003. DESIGN AND SETTING: This was a population-based descriptive study, Brazil, 2003. METHODS: Data from 126,520 fatal injuries and 733,712 nonfatal injuries seen at public hospitals were analyzed. The data were stratified by sex, age, intent and injury mechanism. Raw and age- and sex-specific rates were calculated per 100,000 individuals. RESULTS: The raw injury mortality rate was 71.5/100,000 (122.6/100,000 for male and 22.0/100,000 for female). For fatal injuries, the proportions of unintentional and intentional injuries were equal (44.3% and 46.9%, respectively). Homicides were the leading cause, 40.3% overall (28.8/100,000), followed by transport-related deaths, 26.2% overall (17.0/100,000). For nonfatal injuries, the rate was 414.8/100,000 and unintentional injuries were predominant (88.9%). Overall, the leading cause was unintentional falls, accounting for 42.6% of victims treated in public hospitals (176.8/100,000). Transport-related injuries were second: 15.0% overall; 62.0/100,000. Fractures comprised 46.7% of principal diagnoses at hospitals. The injury types in the fatal and nonfatal datasets varied according to sex and age. The highest rates were found among young males and elderly people. CONCLUSIONS: Injury prevention activities need to be developed. To prevent deaths, homicide has to be addressed. Among hospitalized cases, falls are the most important problem. Traffic-related injuries play an important role in morbidity and mortality. PMID- 17086303 TI - Climacteric complaints among very low-income women from a tropical region of Brazil. AB - CONTEXT AND OBJECTIVE: Climacteric symptoms may vary between different countries and cultures. Socioeconomic factors and climate may be implicated. The aim of this study was to identify climacteric symptomatology among very low-income Brazilian women, living in a hot and humid region. DESIGN AND SETTING: This cross sectional population-based study was conducted in Cuiaba, at Julio Muller University Hospital, a tertiary institution. METHODS: The study enrolled 354 climacteric women. The variables analyzed were social class, symptomatology and abnormal concurrent conditions. The study was approved by the hospital's research ethics committee. RESULTS: Sixty-five percent of the participants (232/354) were very poor and had had little schooling. The number of symptoms per woman was 8.0 +/- 5.7. Hot flushes, nervousness, forgetfulness and fatigue were each found in nearly 60.0%. Tearfulness, depression, melancholy and insomnia were also frequent. Sexual problems were reported by 25%. The most relevant concurrent abnormal conditions reported were hypertension (33.9%), obesity (26.5%), arthritis/arthrosis (15.0%) and diabetes mellitus (9.6%). Hot flushes were associated with tearfulness, nervousness and forgetfulness. CONCLUSION: Brazilian climacteric women of low income and low schooling present multiple symptoms. Vasomotor and psychosexual symptoms were the most prevalent disorders. Hot flushes were associated with nervousness, forgetfulness and tearfulness. PMID- 17086304 TI - Effect of metabolic control on the in vitro proliferation of peripheral blood mononuclear cells in type 1 and type 2 diabetic patients. AB - CONTEXT AND OBJECTIVE: Diabetes mellitus is a clinical syndrome that frequently leads to the development of chronic complications and high susceptibility to infections. It is probably due to defective immunological defense, which may be related to metabolic control of the disease. The aim of this study was to evaluate the effect of metabolic control on immune-cell behavior in type 1 and type 2 diabetic patients. For this, the in vitro proliferation of peripheral blood mononuclear cells (PBMC) was analyzed in patients with inadequate and adequate metabolic control. DESIGN AND SETTING: Experimental/laboratory study at a university hospital. METHODS: Eleven type 1 and thirteen type 2 diabetic patients were studied, together with 21 healthy individuals divided in two groups (11/10), who were matched by sex and age with those diabetic patients. PBMC cultures stimulated with concanavalin-A (Con-A) were used to measure 3H-thymidine incorporation after 72 hours of cell culturing. For patients with inadequate metabolic control, culturing was performed on the first day of patient hospitalization and again after intensive treatment to achieve adequate control. RESULTS: The proliferation index for Con-A-stimulated cultures from type 1 diabetic patients was significantly greater than that for cultures from healthy individuals and type 2 diabetic patients, independent of metabolic control. A negative correlation between the proliferation cell index and body mass index and serum C-reactive protein levels was also observed. CONCLUSION: The increase in the proliferation capacity of type 1 diabetic T lymphocytes was probably not caused by hyperglycemia and/or insulinopenia related to inadequate metabolic control. PMID- 17086305 TI - Positive correlation between serum and peritoneal fluid CA-125 levels in women with pelvic endometriosis. AB - CONTEXT AND OBJECTIVE: One of the diagnostic markers of endometriosis is CA-125, and elevated levels of this are caused by high concentrations in the ectopic endometrium. The objective of this study was to correlate CA-125 levels in serum and peritoneal fluid from women with and without pelvic endometriosis. DESIGN AND SETTING: This was a prospective, cross-sectional, controlled study of consecutive patients undergoing laparoscopy for infertility, pelvic pain or tubal ligation, during early follicular phase, at the university hospital of Faculdade de Medicina de Ribeirao Preto. METHODS: Fifty-two patients were divided into two groups: endometriosis group, consisting of 35 patients with biopsy-confirmed pelvic endometriosis, and control group, consisting of 17 patients without endometriosis. CA-125 levels in serum samples and peritoneal fluid were determined by chemiluminescence. RESULTS: CA-125 levels in serum and peritoneal fluid were higher in patients with advanced pelvic endometriosis (means of 39.1 +/- 45.8 U/ml versus 10.5 +/- 5.9 U/ml in serum, p < 0.005; 1,469.4 +/- 1,350.4 U/ml versus 888.7 +/- 784.3 U/ml in peritoneal fluid, p < 0.05), and showed a positive correlation between each other (correlation coefficient (r) = 0.4880). Women with more advanced degrees of endometriosis showed higher CA-125 levels in both serum and peritoneal fluid (p = 0.0001). CONCLUSION: There is a positive correlation between serum and peritoneal fluid values of CA-125 in women with and without endometriosis, and their levels are higher in peritoneal fluid. Advanced endometriosis is related to higher levels in both serum and peritoneal fluid. PMID- 17086306 TI - Research publications in the field of health: omission of hypotheses and presentation of common-sense conclusions. AB - CONTEXT AND OBJECTIVE: Medical literature should consist of knowledge applicable to professional education; nevertheless, the profusion of articles in databases provokes disquiet among students. The authors considered the premise that scientific production in the field of health follows a mechanical description of phenomena without the clarity of motivating questions. The aim was to interpret material from expert reports, applied by medical students to analyze articles from renowned journals. DESIGN AND SETTING: This research project was exploratory, searching for latent meanings regarding methodological problems in a sample of papers. It was performed in a Brazilian medical school. METHODS: The sample was intentionally built, consisting of articles related to original research in the field of health, published over the previous five years. The results came from text content analysis, performed by a professor and his medical students. RESULTS: (1) Failure to state a hypothesis is an equivocal practice: articles did not show clarity of hypothesis to demonstrate that their authors had epistemological knowledge of the methods chosen. (2) There is a certain belief that in normal scientific practice, hypotheses are unnecessary: studies without explicit hypotheses led to suppositions that they merely repeat dominant models. (3) Presentation of common sense as scientific conclusions: research brings together what would have mobilized the researchers initially. CONCLUSIONS: Absence of formal hypotheses leaves scientific production vulnerable when put under epistemological discussion. Conclusions from scientific articles are often confounded with common-sense statements. Quantitative research is suggested, for studying the frequency of occurrence of these dubious methodological points. PMID- 17086307 TI - Spontaneous cholecystocutaneous fistula: a rare complication of gallbladder disease. AB - CONTEXT: Spontaneous cholecystocutaneous abscess or fistula is an extremely uncommon complication secondary to cholecystitis. Over the past 50 years fewer than 20 cases of spontaneous cholecystocutaneous fistulas have been described in the medical literature. We here report a case of subcutaneous gallstone as a rare clinical presentation of the already uncommon cholecystocutaneous fistula. CASE REPORT: An 81-year-old man presented with a large subcutaneous abscess in the right subcostal area with surrounding cellulitis and crepitus. An abdominal computed tomography scan showed two subcutaneous gallstones and communication between the abscess and the gallbladder. Cholecystectomy was performed and the abdominal wall abscess was drained externally. This case report demonstrates that maintaining a high degree of suspicion of this rare entity is helpful in achieving correct preoperative diagnosis, and that computed tomography scan should be performed in all cases of unexplained abdominal wall suppuration or cellulitis. PMID- 17086308 TI - Brazilian medical students want to learn homeopathy and acupuncture in medical schools. PMID- 17086309 TI - PCR-based identification of Burkholderia pseudomallei. AB - DNA amplification techniques are being used increasingly in clinical laboratories to confirm the identity of medically important bacteria. A PCR-based identification method has been in use in our centre for 10 years for Burkholderia pseudomallei and was used to confirm the identity of bacteria isolated from cases of melioidosis in Ceara since 2003. This particular method has been used as a reference standard for less discriminatory methods. In this study we evaluated three PCR-based methods of B. pseudomallei identification and used DNA sequencing to resolve discrepancies between PCR-based results and phenotypic identification methods. The established semi-nested PCR protocol for B. pseudomallei 16-23s spacer region produced a consistent negative result for one of our 100 test isolates (BCC #99), but correctly identified all 71 other B. pseudomallei isolates tested. Anomalous sequence variation was detected at the inner, reverse primer binding site for this method. PCR methods were developed for detection of two other B. pseudomallei bacterial metabolic genes. The conventional lpxO PCR protocol had a sensitivity of 0.89 and a specificity of 1.00, while a real-time lpxO protocol performed even better with sensitivity and specificity of 1.00, and 1.00. This method identified all B. pseudomallei isolates including the PCR negative discrepant isolate. The phaC PCR protocol detected the gene in all B. pseudomallei and all but three B. cepacia isolates, making this method unsuitable for PCR-based identification of B. pseudomallei. This experience with PCR-based B. pseudomallei identification methods indicates that single PCR targets should be used with caution for identification of these bacteria, and need to be interpreted alongside phenotypic and alternative molecular methods such as gene sequencing. PMID- 17086310 TI - Pathogenicity of Entamoeba dispar under xenic and monoxenic cultivation compared to a virulent E. histolytica. AB - Two xenic isolates and cloned cultures of Entamoeba dispar were submitted to monoxenization using Crithidia fasciculata as the associated organism. Growth in monoxenic cultivation and ability of xenic and monoxenic trophozoites to destroy VERO cells and produce lesions in hamster livers were compared to those of a virulent E. histolytica. Parental and cloned E. dispar under monoxenic cultivation showed a remarkable lower growth than the monoxenic E. histolytica and were avirulent in both in vivo and in vitro tests. When xenically cultured, trophozoites of E. dispar showed a moderate lytic activity against VERO cells (1.5 to 41.8% of destruction) but caused severe hepatic lesions in hamsters as those caused by the virulent E. histolytica (29 to 100% in prevalence and 0.86 to 4.00 in lesion degree). Although E. dispar has not been associated with invasive disease in men, the ability of xenic trophozoites to produce prominent tissue damage in experimental conditions has indicated that some strains have a considerable pathogenic potential when in presence of bacteria. PMID- 17086311 TI - Frequency and risk factors for toxocariasis in children from a pediatric outpatient center in southeastern Brazil. AB - To estimate the frequency of anti-Toxocara sp. antibodies, and evaluate factors associated with this infection, sera from 242 male and female children, aged from one to fifteen years old, attended at the Hospital of the Federal University of Uberlandia, Minas Gerais State, Brazil, were analyzed by ELISA. Information on the patients was collected and registered using an investigative questionnaire, and details on possible clinical alterations were obtained from the medical charts of 187 patients. Of a total of 242 samples, 21 (8.7%) were positive for anti-Toxocara sp. antibodies. The presence of dogs and cats and the school variable (place of contact), appeared to be significantly associated (p<0.05) with a positive serology. Respiratory symptoms and eosinophil counts greater than 20% also showed a positive statistical correlation with a positive serology for Toxocara sp.. Factors such as sex and age, and symptoms like headache, stomach ache, convulsive crises and anemia were not associated with toxocariasis. PMID- 17086312 TI - Asymptomatic oral carriage of Candida species in HIV-infected patients in the highly active antiretroviral therapy era. AB - Oropharyngeal candidiasis is the most common opportunistic fungal infection in individuals infected with human immunodeficiency virus. CD4+ lymphocytes count and the quantification of viral RNA in blood plasma have been found to be the main markers of HIV disease progression. The present study was conducted to evaluate Candida sp. diversity in the oral cavity of HIV-infected patients and to determine whether there was association of CD4+ cell count and viral load with asymptomatic oral Candida carriage. Out of 99 HIV-positive patients studied, 62 (62.6%) had positive culture for Candida (oral carriage) and 37 patients (37.4%) had Candida negative culture (no oral carriage). The etiologic agents most common were C. albicans and C. tropicalis. The range of CD4+ was 6-2305 cells/mm3 in colonized patients and 3-839 cells/mm3 for non-colonized patients, while the viral load was 60-90016 copies/mL for colonized patients and 75-110488 copies/mL for non colonized patients. The viral load was undetectable in 15 colonized patients and in 12 non colonized patients. Our results showed that there was no significant difference of the variables CD4+ cell count and viral load between oral candida carriage and no oral candida carriage patients. PMID- 17086313 TI - The present situation regarding lymphatic filariasis in Cabo de Santo Agostinho, Pernambuco, Northeast Brazil. AB - A cross-sectional population-based survey on the occurrence of lymphatic filariasis was carried out in the municipality of Cabo de Santo Agostinho, Pernambuco, Northeast Brazil. 7,650 individuals of both sexes were examined (from 1,416 households) of whom six tested positive for microfilaria according to the thick blood diagnostic test. The age of the individuals examined varied from 0 to 98 (averaging 26.6 years), whilst the age of the microfilaria-positive individuals varied from 11 to 29, averaging 22.5 years. Five of the six positive cases were male. These cases were residents of the following areas: Pista Preta (one case); Ponte dos Carvalhos (four cases); and Pontezinha (one case). This last case from Pontezinha was autochthonous. Of the individuals examined, 109 (1.4%) cited complaints relating to filariasis. These results suggest that filariasis is being transmitted in the municipality of Cabo de Santo Agostinho, a finding that establishes the need to carry out disease control activities, and to collaborate with the planning of the national programme for the elimination of filariasis. PMID- 17086314 TI - Occurrence of enteroparasites in day care centers in Botucatu (Sao Paulo State, Brazil) with emphasis on Cryptosporidium sp., Giardia duodenalis and Enterobius vermicularis. AB - The occurrence of the enteroparasites was verified in 279 children (0 to 6 years) of four municipal day cares of Botucatu/SP. Three samples of each child's feces were collected and processed by the methods of Hoffman-Pons-Janner, Faust and Ritchie and subsequent coloration of the fecal smear by the methods of Auramina-O and Ziehl-Neelsen modified for diagnosis of Cryptosporidium sp. and Graham method for diagnosis of Enterobius vermicularis. Of the analyzed children we verified a prevalence of intestinal parasitism in 53.40%, and the most frequent parasite was Giardia duodenalis (26.88%). Significant association was verified among enteroparasitosis, family income, maternal education and age; the lowest enteroparasite frequency occurred in children of families with larger income and higher education. It was observed that G. duodenalis is more prevalent in children from 0 to 4 years and E. vermicularis is more frequent in children between three and four years old. The high enteroparasite prevalence in day cares suggests complex structure in its epidemiology, where factors beyond sanitation should be considered. PMID- 17086316 TI - Comparison of serology, antigenemia assay and the polymerase chain reaction for monitoring active cytomegalovirus infections in hematopoietic stem cell transplantation patients. AB - Forty-six allogeneic hematopoietic stem cell transplantation (HSCT) patients were monitored for the presence of CMV antibodies, CMV-DNA and CMV antigens after transplantation. Immunoenzymatic serological tests were used to detect IgM and the increase in CMV IgG antibodies (increase IgG), a nested polymerase chain reaction (N-PCR) was used to detect CMV-DNA, and an antigenemia assay (AGM) was used to detect CMV antigens. The presence of CMV-IgM and/or CMV-increase IgG antibodies was detected in 12/46 (26.1%) patients, with a median time between HSCT and the detection of positive serology of 81.5 days. A positive AGM was detected in 24/46 (52.2%) patients, with a median time between HSCT and antigen detection of 62 days. Two or more consecutive positive N-PCR results were detected in 32/46 (69.5%) patients, with a median time between HSCT and the first positive PCR of 50.5 days. These results confirmed that AGM and mainly PCR are superior to serology for the early diagnosis of CMV infection. Six patients had CMV-IgM and/or CMV-increase IgG with a negative AGM (five cases) or N-PCR assay (one case). In five of these cases the serological markers were detected during the first 100 days after HSCT, the period of highest risk. These findings support the idea that serology may be useful for monitoring CMV infections in HSCT patients, especially when PCR is unavailable. PMID- 17086317 TI - Human papillomavirus status and cervical abnormalities in women from public and private health care in Rio de Janeiro State, Brazil. AB - This article reports the HPV status and cervical cytological abnormalities in patients attended at public and private gynecological services from Rio de Janeiro State. It also comments the performance of each HPV DNA tests used. A set of 454 women from private health clinics was tested by routine Capture Hybrid II HPV DNA assay. Among these, 58.4% presented HPV and nearly 90% of them were infected by high risk HPV types. However, this group presented few premalignant cervical lesions and no invasive cervical cancer was registered. We also studied 220 women from low income class attended at public health system. They were HPV tested by polymerase chain reaction using My09/11 primers followed by HPV typing with E6 specific primers. The overall HPV prevalence was 77.3%. They also showed a high percentage of high squamous intraepithelial lesion-HSIL (26.3%), and invasive cervical carcinoma (16.3%). HPV infection was found in 93.1% and 94.4% of them, respectively. The mean ages in both groups were 31.5 and 38 years, respectively. In series 1, HPV prevalence declined with age, data consistent with viral transient infection. In series 2, HPV prevalence did not decline, independent of age interval, supporting not only the idea of viral persistence into this group, but also regional epidemiological variations in the same geographic area. Significant cytological differences were seen between both groups. Normal and benign cases were the most prevalent cytological findings in series 1 while pre-malignant lesions were the most common diagnosis in the series 2. HPV prevalence in normal cases were statistically higher than those from series 1 (p<0.001), indicating a higher exposure to HPV infection. Women from both samples were referred for previous abnormal cytology. However, socio demographic evidence shows that women from series 1 have access to treatment more easily and faster than women from series 2 before the development of pre malignant lesions. These data provides baseline support for the role of social inequalities linked to high risk HPV infection leading to cervical cancer. Broadly screening programs and the development of safe and effective vaccines against HPV would diminish the toll of this disease that affect mainly poor women. PMID- 17086319 TI - Survival of Trypanosoma cruzi in sugar cane used to prepare juice. AB - Chagas disease can be transmitted to man by many different means, including contact with infected triatomine feces, blood transfusion, laboratory accidents, organ transplants, and congenital or oral routes. The latter mode has received considerable attention recently. In this assay, we evaluate the survival of Trypanosoma cruzi contaminating sugar cane used to prepare juice, as well as the viability and capacity for infection by the parasite after recovery. Thirty triatomines were contaminated with T. cruzi Y strain and 45 days later pieces of sugar cane were contaminated with the intestinal contents of the insects. The pieces were ground at different intervals after contamination (time=0, 1, 4, 6, 12 and 24 hours) and the juice extracted and analyzed. Different methods were used to show T. cruzi in the juice: direct analysis, hematocrit tube centrifugation and QBC, and experimental inoculation in 47 female BALB/c mice (five control mice and seven mice for each interval examined (five inoculated orally and two intraperitoneally). Positive results were found using the direct analysis and QBC methods for juice prepared up to 12 hours after initial contamination. However, by the centrifugation technique, positivity was found only up to four hours after contamination of the sugar cane. Inoculated animals showed parasitemia during a 14 day observation period, demonstrating the high survival rate of T. cruzi in sugar cane. PMID- 17086320 TI - Listeria monocytogenes peritonitis in cirrhotic patients: first description in Brazil. AB - Two cases of spontaneous bacterial peritonitis (SBP) caused by Listeria monocytogenes in cirrhotic patients are reported. In one of the cases, the microorganism was isolated from pleural effusion and ascites. SBP is a serious and common complication of patients with ascites caused by hepatic cirrhosis and the culture of the ascitic fluid is an important tool for the diagnosis and for the more appropriate treatment. Although a third generation cephalosporin has usually been employed for empiric treatment of SBP, it does not provide adequate coverage against Listeria spp. In such cases the use of ampicillin (with or without sulbactam) or sulfamethoxazole-trimethoprim is recommended. The last one is used for secondary prophylaxis, instead of norfloxacin. To summarize, Listeria monocytogenes infection is a rare cause of SBP, whose treatment should be specific for the bacteria. PMID- 17086321 TI - Rhinoscleroma: eight Peruvian cases. AB - Rhinoscleroma is a rare infection in developed countries; although, it is reported with some frequency in poorer regions such as Central Africa, Central and South America, Eastern and Central Europe, Middle East, India and Indonesia. Nowadays, rhinoscleroma may be erroneously diagnosed as mucocutaneos leishmaniasis, leprosy, paracoccidioidomycosis, rhinosporidiasis, late syphilis, neoplasic diseases or other upper airway diseases. From 1996 to 2003, we diagnosed rhinoscleroma in eight patients attended in the Dermatologic and Transmitted Diseases service of "Cayetano Heredia" National Hospital, in Lima, Peru. The patients presented airway structural alterations producing nasopharyngeal, oropharyngeal and, in one patient, laryngeal stenosis. Biopsy samples revealed large vacuolated macrophages (Mikulicz cells) in all patients. Ciprofloxacin 500 mg bid for four to 12 weeks was used in seven patients and oxytetracycline 500 mg qid for six weeks in one patient. After follow-up for six to 12 months the patients did not show active infection or relapse, however, all of them presented some degree of upper airway stenosis. These cases are reported because of the difficulty diagnosing the disease and the success of antibiotic treatment. PMID- 17086322 TI - Candida albicans skin abscess. AB - Subcutaneous candidal abscess is a very rare infection even in immunocompromised patients. Some cases are reported when breakdown in the skin occurs, as bacterial cellulites or abscess, iatrogenic procedures, trauma and parenteral substance abuse. We describe a case of Candida albicans subcutaneous abscess without fungemia, which can be associated with central venous catheter. PMID- 17086323 TI - Sea urchin granuloma. AB - Injuries caused by venomous and poisonous aquatic animals may provoke important morbidity in humans. The phylum Echinoderma include more than 6000 species of starfish, sea urchins, sand dollars, and sea cucumbers some of which have been found responsible for injuries to humans. Initial injuries by sea urchins are associated with trauma and envenomation, but later effects can be observed. Sea urchin granuloma is a chronic granulomatous skin disease caused by frequent and successive penetration of sea urchin spines which have not been removed from wounds. The authors report a typical case of sea urchin granuloma in a fisherman and its therapeutic implications. PMID- 17086324 TI - [Building bridges between research and policy to extend social protection in health in Latin America and the Caribbean: a strategy for cooperation]. AB - The Pan American Health Organization (PAHO) and International Development Research Centre (IDRC) have promoted a joint initiative to design, implement, and evaluate innovative strategies for the Extension of Social Protection in Health (SPH) in Latin America and the Caribbean (LAC), involving active partnership between researchers and research users. This initiative was based on a previous review of research on health sector reforms and the recommendations of the workshop on "Health Sector Reforms in the Americas: Strengthening the Links between Research and Policy" (Montreal, Canada, 2001). In its first phase, the initiative supported the development of proposals aiming to extend SPH, elaborated jointly by researchers and decision-makers. In the second phase, the implementation of five of these proposals was supported in order to promote the development of new SPH strategies and new stakeholder interaction models. In this edition of the journal, the process of linking researchers and decision-makers will be analyzed in the context of the five projects supported by this initiative. PMID- 17086336 TI - [Bridging gaps between research and policy]. PMID- 17086335 TI - [Governance in health: a conceptual and analytical approach to research]. AB - In the Latin American region there is a notable absence of conceptual coherency in the use of the terms governability and governance. This is true for their application to both the social and political sciences and health. Researchers' understanding of governance varies, and the concept is used heterogeneously within academic circles, with ignorance of the term on the part of decision makers and great confusion and ambiguity in the meanings used by researchers and decision-makers in the health sector. Instead of the prevailing normative use, promoted by most international agencies, a conceptual and analytical framework for governance is proposed here for health systems and services research. Advances in the design of this framework were used to evaluate the public health insurance program in Buenos Aires, Argentina, which allowed the use of the analytical framework to be assessed as a tool for systemizing the social, political, and institutional complexity of the health policy formulation and implementation processes. PMID- 17086337 TI - [Interaction between researchers and decision-makers: a case study]. AB - This article analyzes the process of interaction between the Instituto de la Salud Juan Lazarte research team and the Buenos Aires Public Health Insurance (PHI) management team during the design and development of the study titled "PHI Institutional Capability Analysis and Performance Evaluation", currently underway. From a cross-disciplinary perspective, examining different areas of interaction between the SPS management team and the research team, the relationship is characterized as an application of the "interaction model". This approach promotes the construction of interfaces that allow the development of negotiation and collaboration between the scientific and political "communities". Application of this model has produced changes in the conceptual and methodological framework and in substantive issues during implementation of the SPSMI. PMID- 17086338 TI - [Research for decision-making: a case study in Brazil]. AB - This article analyzes the experience of the relationship between researchers and decision-makers in a study based on the hypothesis that this relationship would favor the incorporation of research results in health policy implementation. The attempt was made to identify elements that affect the relationship according to different policy and research phases, with an emphasis on the appropriation of the results by the decision-makers. It was shown that this research model per se does not guarantee the incorporation of the results, since variables related to the political/institutional context, the decision-makers' profile, and the forms of relationship played a preponderant role in this case. In research oriented towards decision-making, the use of the results depends on the decision-making timing, the capacity for dialogue, the policy's institutionality, and the decision-makers' place in the overall political context. PMID- 17086339 TI - Linking researchers and policy-makers: some challenges and approaches. AB - In developing a proposal for the study of the effect of user fees on access to preventive care, a team, comprising researchers and policy-makers, initiated interaction with key policy implementers in the Jamaica's Ministry of Health to ensure that their perspectives were considered at the preliminary stage. There were many pressing events occupying the minds and energies of the implementers, but the team was able to capitalize on existing good relationships to capture attention. In the interviews that followed, agreement was reached on the necessity for the study, its focus and methodology. The process of consultation achieved notable successes and can be regarded as a model for successful research and policy interaction. PMID- 17086340 TI - Use of research results in policy decision-making, formulation, and implementation: a review of the literature. AB - This paper offers a critical review of the theoretical literature on the relationship between the production of scientific knowledge and its use in policy formulation and implementation. Extensive academic output, using a diversity of approaches and analytical frameworks, has sought to systematize knowledge transfer categories and strategies with a view to improving the application of scientific knowledge. A considerable part of this thinking addresses the problem from a more traditional perspective, which (explicitly or implicitly) regards research results as an "accumulable product", depicts the decision-making process simplistically and linearly, and thus restricts strategies to the suiting of research endeavors to needs. Newer approaches place greater importance on the complexity of policymaking and the knowledge production process, which are integrated into and explained in particular political and institutional settings. Although the application of knowledge transfer ideas to health policy and systems research does generate some interesting contributions, a long process of theoretical thinking lies ahead. PMID- 17086341 TI - [Beyond dissemination: lessons from the interaction between researchers and decision-makers during a research project in Bogota, Colombia]. AB - This paper discusses the experiences and conceptual and theoretical lessons learned during interaction between researchers, policy-makers, program beneficiaries, and other actors in a research project on social protection, health, and forced displacement in Bogota, Colombia. The article begins by presenting the methodological approach and a description of interaction between various actors. This experience provides the basis for a conceptual discussion on the factors and determinants of research use. The article also highlights the need for taking an epistemological view beyond the positivist and rationalist underpinnings of the dominant explanations concerning interaction between researchers and decision-makers. PMID- 17086342 TI - [An innovative proposal for health insurance policies based on research results: the case of unemployed in Medellin, Colombia]. AB - In recent decades there has been an important academic debate on how to transfer research-based knowledge to the health policy-making process. Despite an intensive controversy and various approaches to the problem, there is basic consensus concerning both the academic relevance and the usefulness of further evaluation of concrete experiences that allow conclusions and clarification of what are still confusing issues. The current article discusses a joint research experience by professors and local and regional health policy-makers. The research focused on the socioeconomic conditions, duration of the employment/unemployment cycle, and features of health insurance for workers that lost their jobs in the Greater Metropolitan "Valle de Aburra" Area (Antioquia, Colombia) in 2005, as the basis for a proposal to guarantee the maintenance of health insurance for these workers. The potentialities and difficulties of the interactive process are analyzed. The article is also intended to elucidate how scientific outcomes influence policymaking in health, as well as the difficulties imposed by concrete reality and the political process for implementing the innovative proposal. PMID- 17086343 TI - Cross-sector learning among researchers and policy-makers: the search for new strategies to enable use of research results. AB - This paper assesses the preliminary results of a research funding strategy that alters the structure and process of research by requiring interaction between researchers and policy-makers. The five research teams focused on different aspects of expanding social protection in health in Latin America and the Caribbean. Preliminary results revealed negotiation of the research questions at the start of the process, influencing not only the project design, but the decision-makers' ways of thinking about the problem as well. As the projects advanced, turnover among government officials on four of the teams impaired the process. However, for the one team that escaped re-composition, the interaction has led to use of data in decision-making, as well as a clear recognition by both parties that different kinds of evidence were at play. The process highlighted the importance of stimulating systems of learning in which multiple kinds of knowledge interact. This interaction may be a more realistic expectation of such initiatives than the original goal of "transferring" research knowledge to policy and practice. PMID- 17086344 TI - The diagnosis and treatment of infantile nystagmus syndrome (INS). AB - The successful treatment of infantile nystagmus syndrome (INS) depends primarily on accurate and repeatable diagnosis of the type(s) of nystagmus present as well as their variation with gaze and convergence angles or fixating eye. Research over the past 40 years has demonstrated that the only way to achieve both is by making and analyzing ocular motility recordings. Determination of the direct effects of peripheral and central INS therapies can only be made by pre- and post therapy comparisons of the nystagmus characteristics, specifically of the quality of the foveation periods within each cycle. If one is only interested in cosmetic improvements, diminution of the nystagmus amplitude is all that need be measured. However, if improvement of visual function is the primary goal of therapy, then measurement of the pre- and post-therapy foveation quality must be made, both in primary position and over a broad range of gaze angles. The use of the eXpanded Nystagmus Acuity Function (NAFX) on nystagmus data yields both an accurate measure of foveation quality and a prediction of maximum potential acuity for the patient's waveform. When used with the patient's measured, pre-therapy visual acuity, the NAFX demonstrates the amount of visual acuity loss that is due to sensory abnormalities, demonstrates the amount due to the nystagmus waveform, and estimates the measured post-therapy acuity for all values of improved NAFX and gaze angles measured. The ability to predict visual acuity improvement was not possible before the use of the NAFX. The failure to incorporate accurate measures of nystagmus waveform and foveation quality into their diagnostic evaluation continues to deprive patients of the best possible standard of care and results in mistaken diagnoses as well as inappropriate and, in some cases, unneeded multiple surgeries. PMID- 17086345 TI - Combined dexamethasone suppression-corticotropin-releasing hormone stimulation test in studies of depression, alcoholism, and suicidal behavior. AB - The hypothalamic-pituitary-adrenal (HPA) axis controls the secretion of corticotropin-releasing hormone (CRH), corticotropin (adrenocorticotropic hormone, ACTH), and cortisol. The dexamethasone suppression test (DST) is the most frequently used test to assess HPA system function in psychiatric disorders. Patients who have failed to suppress plasma cortisol secretion, i.e., who escape from the suppressive effect of dexamethasone, have a blunted glucocorticoid receptor response. After CRH became available for clinical studies, the DST was combined with CRH administration. The resulting combined dexamethasone suppression-corticotropin-releasing hormone stimulation (DST-CRH) test proved to be more sensitive in detecting HPA system changes than the DST. There is a growing interest in the use of the DEX-CRH test for psychiatric research. The DEX CRH test has been used to study different psychiatric conditions. Major depression, alcoholism, and suicidal behavior are public health problems around the world. Considerable evidence suggests that HPA dysregulation is involved in the pathogenesis of depressive disorders, alcoholism, and suicidal behavior. Over the past 2 decades, there has been a shift from viewing excessive HPA activity in depression as an epiphenomenon to its having specific effects on symptom formation and cognition. The study of HPA function in depression, alcoholism, and suicidal behavior may yield new understanding of the pathophysiology of these conditions, and suggest new approaches for therapeutic interventions. The combined DEX-CRH test may become a useful neuroendocrinological tool for evaluating psychiatric patients. PMID- 17086346 TI - Risk and protective factors for suicide in patients with alcoholism. AB - Alcoholism is associated with a high risk for suicidal behavior. Up to 40% of persons with alcoholism attempt suicide at some time and 7% end their lives by committing suicide. Risk factors include being male, older than 50 years of age, living alone, being unemployed, poor social support, interpersonal losses, continued drinking, consumption of a greater amount of alcohol when drinking, a recent alcohol binge, previous alcohol treatment, a family history of alcoholism, a history of comorbid substance abuse (especially cocaine), a major depressive episode, serious medical illness, suicidal communication, and prior suicidal behavior. Suicidal behavior is especially frequent in patients with comorbid alcoholism and major depression. However, all patients with alcoholism should be evaluated for suicide risk. Understanding of risk and vulnerability to suicidal behavior in alcoholism still outweighs our knowledge of protective factors and resilience. Knowledge of protective factors for suicide may help to prevent and/or predict suicidal behavior. Protective factors for suicide in alcoholism are quite varied and include an individual's biological and behavioral characteristics, as well as attributes of the environment and culture. Protective factors include effective clinical care for psychiatric (including alcoholism and drug abuse) and physical disorders, easy access to a variety of clinical interventions and support for seeking help, restricted access to highly lethal means of suicide, strong connections to family and community support, skills in problem solving and conflict resolution, cultural and religious beliefs that discourage suicide and support self-preservation. Future studies are necessary to determine which interventions may reduce suicidal behavior in alcoholism. PMID- 17086347 TI - [The current revision of Directive 86/609/EWG from the perspective of animal welfare legislation]. AB - The necessity to revise Directive 86/609/EWG has been discussed since 1993, growing more urgent since the Protocol to the Treaty of Amsterdam (1997) and the Treaty establishing a Constitution for the European Union (2004), both of which recognise the welfare of animals as sentient beings; a revision also proves necessary because of various changes within the framework of animal experimentation. Within an expert questionnaire launched in the summer of 2006 several shortcomings of Directive 86/609/EWG are pointed out; proposals for revision mainly concern the scope of the directive, authorisation of animal experiments by national bodies, controls on institutional level, minimal requirements for housing and care of laboratory animals as well as for qualification of staff, and instruments for reducing duplication of animal experiments. The introduction of an obligatory ethical review as part of a standardised and project-related authorisation procedure, the up-grading of housing and care standards and the implementation of mechanisms for reducing duplication of animal experiments may enhance full exploitation of the 3R and are therefore crucially important for a directive complying with the requirements of the Treaty of Amsterdam. PMID- 17086348 TI - Towards eliminating the use of animals for regulatory required vaccine quality control. AB - Traditionally, regulatory required vaccine quality control relies on the use of laboratory animals. Both batch testing for safety and testing for potency are based on animal models. Quite often these models include procedures that induce severe pain and suffering. This has urged the development of 3Rs methods. Some examples of recent achievements in the quality control of toxoid vaccines are the replacement of challenge procedures by serological methods, the reduction of numbers of animals required by changing from multi-dose to single dose testing, and developments in the area of in vitro models and physicochemical techniques. Central in the progress towards the elimination of animal use is the acceptance of the so-called consistency approach. The essence of the consistency approach is that a new batch of vaccine is no longer seen as a unique product but as only one of a series of batches produced from the same starting material (seed lot). Consequently, the new batch shares many of the characteristics of the previous batches produced from the same seed lot. This allows for a new strategy of vaccine quality control consisting of demonstrating consistency in production, placing emphasis on aspects such as in-process testing, and implementing Good Manufacturing Practice and Quality Assurance (QA). The new strategy particularly focuses on non-animal test models such as physicochemical methods and in vitro models. Implementation of the consistency approach might significantly contribute towards the elimination of the use of animals in regulatory required vaccine quality control. PMID- 17086349 TI - Proposal for the composition of animal experiments committees in the Netherlands. AB - The Dutch Act on Animal Experimentation (1996) requires that local animal experiments committees (AECs) review animal experiments and balance the scientific and societal benefits of the experiments against the suffering caused to the animals used. Each AEC is composed of at least seven members who provide a balance of expertise in animal experiments, alternatives to laboratory animal experiments, ethics, and animal welfare and protection. This study proposes selection criteria for individuals possessing each of the four AEC required areas of expertise. Criteria were established minding that, on the one hand, sufficient knowledge and expertise can be demonstrated whilst, on the other hand, a sufficient number of people would qualify to participate in the AECs. The results of this study may serve as a starting point for further discussion of selection criteria for members of AECs both in the Netherlands and in other countries where ethical review processes have been or are being implemented. PMID- 17086350 TI - Computer-based alternatives in higher education--past, present and future. AB - Many thousands of animals are still used annually in tertiary education despite efforts by enthusiastic teachers to reduce this number by developing and making available to their colleagues a range of alternatives. Technology-based alternatives which support replacement and reduction are at the forefront of these efforts. Persuading teachers to use them is critical and strategies to raise awareness and support the integration of the alternatives into teaching are described. Many of the existing computer-based alternatives were developed in the early 1990s and rapid changes in the technologies used to deliver them have rendered them difficult to use and in some cases obsolete despite the fact that their content and educational design are still valid. A project, using a learning object approach to development, which aims to preserve the content and educational design, improve the flexibilty of delivery and enable teachers to edit the content, is also described. PMID- 17086351 TI - [The isolated perfused porcine kidney model for investigations concerning surgical therapy procedures]. AB - Experiments to develop innovative surgical therapy procedures are conventionally conducted on animals, as crucial aspects like tissue removal and bleeding disposition cannot be investigated in vitro. Extracorporeal organ models however reflect these aspects and could thus reduce the use of animals for this purpose fundamentally in the future. The aim of this work was to validate the isolated perfused porcine kidney model with regard to its use for surgical purposes on the basis of histological and radiological procedures. The results show that neither storage nor artificial perfusion led to any structural or functional damage which would affect the quality of the organ. The kidney model is highly suitable for simulating the main aspects of renal physiology and allows a constant calibration of perfusion pressure and tissue temperature. Thus, with only a moderate amount of work involved, the kidney model provides a cheap and readily available alternative to conventional animal experiments; it allows standardised experimental settings and provides valid results. PMID- 17086352 TI - [Austria: fewer experimental animals used in 2005]. AB - The number of animals used in experiments in Austria decreased in 2005 by 10%. This overall positive development must be regarded critically, as the number of big mammals, e.g. cows, used for experimental purposes has increased substantially and as a lot of relevant and necessary data, e.g. on the use of transgenic animals, is still not being collected. The information given in the Austrian statistics on animals used for scientific purposes is inadequate and requires revision. PMID- 17086353 TI - [What can a chair on alternatives to animal experimentation effectuate?]. AB - The DZ foundation has, together with the Thurgau Foundation for Arts and Science (CH) and the state Baden-Wuerttemberg (D), made it possible for the University of Konstanz to establish an endowed chair for In vitro Alternative Methods. Such a step has never been taken before at a German university, and many may ask what the purpose is, and what this should lead to. The foundation document states... "the endowed chair will introduce the 3R principles of moderate and rational animal protection (reduce, replace, refine) into the academic curriculum and the profile of the University in practical and theoretical form". Furthermore, the goal is defined to develop and judge alternative methods to animal experiments in the domain of consumer- and health protection with the help of interdisciplinary research. Is therefore the purpose to establish 3R research in Germany--or make it socially acceptable? Of course, a No is the answer to the first part of the question, as 3R research is already successfully established and practiced at many universities and institutes. The second part of the question touches an interesting point. If one of the most highly-reputed faculties of the state creates a chair to this topic, then there is a clear signaling effect associated with it: in Konstanz 3R research is not seen as a haphazard spin-off, a compulsory exercise during the animal experimentation course, or a recycled use of an old cell culture system, but it is rather seen as an independent subject with high self-esteem and a function in creating a broad awareness. PMID- 17086354 TI - Intraspecies variation in the Kanzawa spider mite differentially affects induced defensive response in lima bean plants. AB - The Kanzawa spider mite, Tetranychus kanzawai, is a polyphagous herbivore that feeds on various plant families, including the Leguminacae. Scars made by the mite on lima bean leaves (Phaseolus lunatus) were classified into two types: white and red. We obtained two strains of mites--"White" and "Red"--by selecting individual mites based on the color of the scars. Damage made by the Red strain induced the expression of genes for both basic chitinase, which was downstream of the jasmonic acid (JA) signaling pathway, and acidic chitinase, which was downstream of the salicylic acid (SA) signaling pathway. White strain mites also induced the expression of the basic chitinase gene in infested leaves but they only slightly induced the acidic chitinase gene. The Red genotype was dominant over the White for the induction of the acidic chitinase gene. The amount of endogenous salicylates in leaves increased significantly when infested by Red strain mites but did not increase when infested by White strain mites. JA and SA are known to be involved in the production of lima bean leaf volatiles induced by T. urticae. The blend of volatiles emitted from leaves infested by the Red strain were qualitatively different from those infested by the White strain, suggesting that the SA and JA signaling pathways are differently involved in the production of lima bean leaf volatiles induced by T. kanzawai of different strains. PMID- 17086355 TI - Tiam1 regulates cell adhesion, migration and apoptosis in colon tumor cells. AB - The guanine nucleotide exchange factor Tiam1 regulates numerous biologic properties including migration and invasion. We demonstrated previously that colon tumor cells biologically selected for increased migration were increased in Tiam1 expression. Cells selected for increased Tiam1 expression or that ectopically overexpress Tiam1 were increased in metastatic potential. Here, we demonstrate that Tiam1 regulates additional functions associated with metastasis, including reduced cellular adhesion and resistance to anoikis. Tiam1 effects on cellular migration are mediated through its downstream substrate, Rac. Increased Tiam1 expression also leads to anoikis-resistance, whereas decreasing Tiam1 expression by siRNA sensitizes cells to this form of apoptosis; however, Tiam1's regulation of anoikis is Rac-independent. Staurosporine sensitivity is also Rac independent, suggesting Tiam1's effects on apoptosis require other effectors. As many of the observed phenotypes are characteristic of a transition of transformed epithelial cells to a mesenchymal-like phenotype, we also examined biochemical properties associated with an EMT. We demonstrate an increase in vimentin expression in cell lines that overexpress Tiam1 and have a more metastatic phenotype. Concomitant with this increase, we observe a decrease in E-cadherin expression in these cells. Lastly, we stained a panel of human colorectal specimens and adjacent normal tissue, and demonstrate that Tiam1 is overexpressed in a subset of human colorectal tumors. In summary, in colon tumor cells, Tiam1 affects multiple properties associated with acquisition of the metastatic phenotype, and may represent a marker of colon tumor progression and metastasis in a subset of tumors. PMID- 17086356 TI - Role of CCR1 and CCR5 in homing and growth of multiple myeloma and in the development of osteolytic lesions: a study in the 5TMM model. AB - Multiple myeloma (MM) is a plasma cell malignancy, characterized by the localization of the MM cells in the bone marrow (BM), where they proliferate and induce osteolysis. The MM cells first need to home or migrate to the BM to receive necessary survival signals. In this work, we studied the role of CCR1 and CCR5, two known chemokine receptors, in both chemotaxis and osteolysis in the experimental 5TMM mouse model. A CCR1-specific (BX471) and a CCR5-specific (TAK779) antagonist were used to identify the function of both receptors. We could detect by RT-PCR and flow cytometric analyses the expression of both CCR1 and CCR5 on the cells and their major ligand, macrophage inflammatory protein 1alpha (MIP1alpha) could be detected by ELISA. In vitro migration assays showed that MIP1alpha induced a 2-fold increase in migration of 5TMM cells, which could only be blocked by TAK779. In vivo homing kinetics showed a 30% inhibition in BM homing when 5TMM cells were pre-treated with TAK779. We found, in vitro, that both inhibitors were able to reduce osteoclastogenesis and osteoclastic resorption. In vivo end-term treatment of 5T2MM mice with BX471 resulted in a reduction of the osteolytic lesions by 40%; while TAK779 treatment led to a 20% decrease in lesions. Furthermore, assessment of the microvessel density demonstrated a role for both receptors in MM induced angiogenesis. These data demonstrate the differential role of CCR1 and CCR5 in MM chemotaxis and MM associated osteolysis and angiogenesis. PMID- 17086357 TI - Detection of Survivin-expressing circulating cancer cells in the peripheral blood of breast cancer patients by a RT-PCR ELISA. AB - Survivin mRNA expression was detected in 69.2%-93.8% of primary breast carcinomas, but is rarely expressed in normal breast tissues and hematopoietic cells. The objective of this study was to investigate the significance that the detection of Survinin-expressing circulating breast cancer cells in the peripheral blood has on clinical outcomes. The detection method was based on a RT PCR ELISA technique developed in our laboratory. Sixty-seven breast cancer patients in various stages and 135 normal healthy women were investigated. Survivin-expressing circulating cancer cells were detected in the peripheral blood samples from 34 (50.7%) out of 67 breast cancer patients, but not in the healthy women that were used as controls. The presence of Survivin-expressing circulating breast cancer cells was found to be significantly associated with various clinicopathological parameters such as vessel infiltration, histological grade, tumor size, nodal status, ER/PgR status, Her-2 status and clinical stages of the disease (P < 0.01). During a follow-up period of 36 months, 9 out of 11 (81.8%) breast cancer patients that had a positive Survivin-expressing at the time of the initial assay test suffered a relapse of the disease, whereas recurrence was only found in 2 out of 6 (33.3%) breast cancer patients that had a negative Survivin-expression. Thus, the detection of circulating cancer cells expressing Survivin mRNA could provide valuable information for the prediction of metastasis and recurrence of breast cancer. PMID- 17086358 TI - Inhibition of adhesion and proliferation of peritoneally disseminated tumor cells by pegylated catalase. AB - Hydrogen peroxide may aggravate the peritoneal dissemination of tumor cells by activating the expression of a variety of genes. In this study, we used pegylated catalase (PEG-catalase) to examine whether prolonged retention of catalase activity within the peritoneal cavity is effective in inhibiting peritoneal dissemination in mouse models. Murine B16-BL6 cells or colon 26 cells labeled with firefly luciferase gene were inoculated intraperitoneally into syngeneic mice. Compared with unmodified catalase, PEG-catalase was retained in the peritoneal cavity for a long period after intraperitoneal injection. A single injection of PEG-catalase just before tumor inoculation significantly reduced the number of the tumor cells at 1 and 7 days. The changes in the expression of molecules involved in the metastasis were evaluated by real time quantitative PCR analysis. Inoculation of the tumor cells increased the expression of intercellular adhesion molecule (ICAM)-1 in the greater omentum, which was inhibited by PEG-catalase. An injection of PEG-catalase at 3 days after tumor inoculation also reduced the number of the tumor cells, suggesting that processes other than the adhesion of tumor cells to peritoneal organs are also inhibited. Daily doses of PEG-catalase significantly prolonged the survival time of tumor bearing mice. These results indicate that intraperitoneal injection of PEG catalase inhibits the multiple processes of peritoneal dissemination of tumor cells by scavenging hydrogen peroxide in the peritoneal cavity. PMID- 17086359 TI - Neutrophil activator of matrix metalloproteinase-2 (NAM). AB - We have isolated a novel soluble factor(s), neutrophil activator of matrix metalloproteinases (NAM), secreted by unstimulated normal human peripheral blood neutrophils that causes the activation of cell secreted promatrix metalloproteinase-2 (proMMP-2). Partially purified preparations of NAM have been isolated from the conditioned media of neutrophils employing gelatin-Sepharose chromatography and differential membrane filter centrifugation. NAM activity, as assessed by exposing primary human umbilical vein endothelial cells (HUVEC) or HT1080 cells to NAM followed by gelatin zymography, was seen within one hour. Tissue inhibitor of metalloproteinase-2 (TIMP-2) and hydroxamic acid derived inhibitors of MMPs (CT1746 and BB94) abrogated the activation of proMMP-2 by NAM, while inhibitors of serine and cysteine proteases showed no effect. NAM also produced an increase in TIMP-2 binding to HUVEC and HT1080 cell surfaces that was inhibited by TIMP-2, CT1746, and BB94. Time-dependent increases in MT1-MMP protein and mRNA were seen following the addition of NAM to cells. These data support a role for NAM in cancer dissemination. PMID- 17086360 TI - Naturally occurring resistance of bone marrow mononuclear and metastatic cancer cells to anticancer agents. AB - Numerous cancer patients fail standard chemotherapy or develop resistance to chemotherapy during the course of treatment. The purpose of this study is to elucidate the overall response of cells obtained from cancer patients and from normal individuals to chemotherapeutic agents. We analysed the chemosensitivity of cancer cells derived from bone marrow and from pleural effusions or ascites fluids from patients with different cancers. Chemosensitivity to doxorubicin, cisplatin and paclitaxel was determined using the MTT assay. We also determined the response of bone marrow mononuclear (BMMN) cells. There was a wide range of responses to chemotherapy drugs in samples from different individuals. This was observed in cells derived from bone marrow and from ascites or pleural fluids. Large variations were also observed among morphologically normal BMMN cells and metastatic cancer cells from chemo-naive patients. Cancer cells can easily be collected from ascites or pleural fluids and reliably assayed for chemosensitivity. We describe here that inherent chemoresistance may be a reason for the lack of response to chemotherapy in some patients. We discuss the potential of using the determination of natural resistance to dictate the drugs to be employed for treatment. PMID- 17086361 TI - Reduction of invasive potential in K-ras-transformed thyroid cells by restoring of TGF-beta pathway. AB - Transforming Growth Factor-beta1 (TGF -beta1) is a multifunctional cytokine that regulates a number of cellular processes such as cell growth, differentiation, plasticity, cell motility, adhesiveness, embryogenesis, development and apoptosis through binding to TGF-beta receptors. We have previously demonstrated that K-ras transformed rat thyroid cells, K10, are resistant to the growth inhibitory action of TGF-beta1, because they show a decreased expression of type II receptor (TbetaRII). Clones obtained transfecting TbetaRII, partially revert their malignant phenotype, showing a reduction in the anchorage-dependent and independent cell growth and a statistically significant decrease in tumourigenicity with respect to the highly malignant parental cells, both in spontaneous and artificial metastases, when transplanted in athymic nude mice. The purpose of the present work is to elucidate the molecular events involved in the modulation of the tumourigenic potential of K-ras-transformed rat thyroid cells overexpressing TbetaRII. Our data demonstrate that the TbetaRII overexpressed in K-ras-transformed thyroid cell clones is a functional receptor and is essential to restore in these cells behaviour similar to that of control cells. The TbetaRII overexpression is responsible for a strong reduction of adhesive and migratory behaviour of highly malignant K-ras-transformed thyroid cells. These results suggest that the restore of a functional TGF-beta receptor in these cells may be useful for the limitation of tumour spread and dissemination. PMID- 17086362 TI - Association of high-density lipoprotein levels and carotid atherosclerotic plaque characteristics by magnetic resonance imaging. AB - A low level of high-density lipoprotein cholesterol (HDL-C) is a risk factor for atherosclerotic disease. Magnetic resonance imaging (MRI) can provide detailed information on carotid atherosclerotic plaque size and composition. The purpose of this study was to correlate HDL levels with carotid plaque burden and composition by MRI. Thirty-four patients with coronary artery disease (CAD) receiving simvastatin plus niacin or placebo for both drugs for three years were randomly selected to undergo MRI of carotid arteries. Atherosclerotic plaque wall volumes (WVs) and plaque components including lipid rich/necrotic core (LR/NC), calcium, fibrous tissue, and loose matrix were measured. Mean WV or atherosclerotic burden was significantly associated with total HDL-C levels (r = 0.39, P = 0.02), HDL(2) (r = -0.36, P = 0.03), HDL(3) (r = -0.34, P = 0.04), and LDL/HDL ratio (r = 0.42, P = 0.02). Plaque lipid composition or LR/NC was significantly associated with HDL(3) (r = -0.68, P = 0.02). Patients with low HDL levels ( 35 mg/dL. Among CAD patients, low HDL-C levels were significantly associated with increased carotid atherosclerotic plaque burden and lipid content by MRI. PMID- 17086363 TI - The benefit of 64-MDCT prior to invasive coronary angiography in symptomatic post CABG patients. AB - PURPOSE: The purpose of this study is to assess the diagnostic accuracy of 64 MDCT in symptomatic patients after CABG and to explore the advantages of the 64 MDCT results on the CAG procedure. MATERIAL AND METHODS: From December 2004 until August 2005, 34 post-CABG patients (29 men, mean age 63.5 +/- 8.5 years) with 69 coronary artery bypass grafts were scanned on a 64-MDCT (Somatom Sensation 64, Siemens AG, Forchheim, Germany) prior to CAG. Angiograms and 64-MDCT images were evaluated for the existence of occlusions or significant stenosis (>or=50% lumen reduction) in bypass grafts and native coronary arteries. RESULTS: 64-MDCT had a sensitivity, a specificity, and a diagnostic accuracy of 100% for occlusion detection. For stenosis detection, sensitivity was 100%, specificity 98.7% and diagnostic accuracy 98.7%. For detecting significant stenosis in native coronary arteries, 64-MDCT had a sensitivity of 80.0%, specificity of 90.8%, and a diagnostic accuracy of 87.1%. Seventeen patients (50.0%) did not need invasive treatment, 14 patients (41.2%) underwent a percutaneous coronary intervention (PCI), and 3 patients (8.8%) underwent surgery. Treatment advice based on 64-MDCT was correct in 88.2% of patients and when 64-MDCT results would have been known 58.8% of diagnostic CAG procedures could have been prevented. CONCLUSION: In conclusion, 64-MDCT has a high diagnostic accuracy in detecting bypass graft stenosis and occlusions, and 64-MDCT based treatment advice was correct in 88.2% of patients. PMID- 17086364 TI - Isolated left pulmonary artery stenosis due to extrinsic compression by intra thoracic tumor: recognition of unusual Doppler flow pattern and correlation with computed tomography. AB - Pulmonary artery compression in adults resulting from tumors is an uncommon condition often associated with poor prognosis. Among the imaging modalities used for diagnosis, the role of trans thoracic echocardiography in identifying secondary pulmonic stenosis due to extrinsic or intrinsic compression and more importantly the physiologic significance has been increasingly recognized. We describe here a case of isolated left pulmonary artery stenosis which was initially suspected based on classic echocardiographic features of obstruction of pulmonary artery and subsequently confirmed by CT imaging. This case illustrates the versatility of trans thoracic echocardiography in diagnosing incidental abnormalities with potential significant consequences. PMID- 17086365 TI - A new ultrasonographic technique for diagnosing deep venous insufficiency- imaging and functional evaluation of venous valves by ultrasonography with improved resolution. AB - This is to demonstrate a new 2D-ultrasonographic technique which enabled clear resolution of deformed valves, visualization of venous reflux and quantitation of valve incompetence. In a 59-year-old Japanese female patient, ultrasonography was done using Aplio, Toshiba Medical Systems Co., Japan, equipped with 8 MHz linear probe capable of differential tissue harmonic imaging to diagnose the cause of her leg edema. Venous ultrasonography using this device at the popliteal venous valve in this patient demonstrated clear view of deformed venous valve and valve separation at one end of valvular agger while the other part of the valve is closed. Color Doppler failed to show venous reflux due to its low velocity. However, the appearance and disappearance of a thrombus-like echo could be imaged using 2D-ultrasonography. In addition, we were able to demonstrate the time course change of valve opening and closing, and quantitate the valve incompetence using M-mode ultrasonography. PMID- 17086366 TI - Successful treatment of polymicrobial multivalve infective endocarditis. Multivalve infective endocarditis. AB - A 57-year-old man presented with triple valve infective endocarditis. There were vegetations on the tricuspid, mitral, and aortic valve. He had multiple complications such as pulmonary abscess, severe anaemia, and congestive heart failure. His general condition was extremely poor. Intensive medical therapy, such as blood transfusion, mechanical ventilation, and inotropic support, allowed him to tolerate surgery. Triple valve replacement was successfully performed without major complication. Vegetation cultures grew Streptococcus bovis and Enterecoccus faecalis. At 1 year follow-up, the patient is alive in NYHA functional class I. PMID- 17086367 TI - GM-CSF activates the Jak/STAT pathway to rescue polymorphonuclear neutrophils from spontaneous apoptosis in young but not elderly individuals. AB - Polymorphonuclear neutrophils (PMN) are the first cells to be recruited to the site of tissular aggression. They have a short-life span and die by spontaneous apoptosis. However, their life span and functional activities can be extended in vitro by a number of proinflammatory cytokines, including the granulocyte macrophage colony stimulating factor (GM-CSF). We have reported that the protective effect of GM-CSF did not occur in PMN of elderly subjects. Data reported here showed that this difference was not due to a change in the expression of the GM-CSF receptor in the PMN of elderly individuals compared to young subjects. Furthermore, we showed here that GM-CSF activated the Janus kinase/signal transducer and activator of transcription (Jak/STAT) pathway and this activation appeared to be maintained for an extended period of time (18 h) playing an important role in the GM-CSF induced delayed PMN apoptosis. In marked contrast, GM-CSF had no effects on Jak2 activation in PMN of elderly individuals. We found that an inhibitor of Jak2 activation (AG490) abolished the protective effect of GM-CSF in PMN from young donors, however had no effect in PMN of elderly subjects. GM-CSF induced a transient activation of STAT3 and STAT5 in PMN of young donors but failed to activate to the same extent these signal transducers in PMN of elderly donors. The levels of proCaspase-3 were reduced in PMN of young donors treated with GM-CSF for 18 h but remained unchanged in PMN of elderly subjects treated under the same conditions compared to the untreated PMN. Our data are consistent with the interpretation that, at least in part (1) the protective effect of GM-CSF against apoptosis results from the activation of the Jak/STAT pathway and (2) decreased rescue from apoptosis in PMN of elderly is related to a failure of GM-CSF to activate this pathway in these cells. PMID- 17086368 TI - Random partition models and exchangeability for Bayesian identification of population structure. AB - We introduce a Bayesian theoretical formulation of the statistical learning problem concerning the genetic structure of populations. The two key concepts in our derivation are exchangeability in its various forms and random allocation models. Implications of our results to empirical investigation of the population structure are discussed. PMID- 17086369 TI - Modelling cell growth and its modulation of the G1/S transition. AB - We present a model for the regulation of the G(1)/S transition by cell growth in budding yeast. The model includes a description of cell size, the extracellular nutrient concentration and a simplified model of the G(1)/S transition as originally reported by Chen et al. [Mol. Biol. Cell 11:369-391, 2000]. By considering cell growth proportional to cell size we show that the cell grows exponentially. In the case where cell growth is considered proportional to the concentration of a sizer protein within the cell, our model exhibits both exponential and linear cell growth for varying parameter values. The effects of varying nutrient concentration and initial cell size are considered in the context of whether progression through the cell-size checkpoint occurs. We consider our results in relation to recent experimental evidence and discuss possible experiments for testing our theoretical predictions. PMID- 17086370 TI - Coreceptor switching in HIV-1 subtype B and subtype C. AB - We use a mathematical model to determine the factors affecting the delayed or rare coreceptor switch in HIV-1 subtype C infected individuals. The model takes into account the two main target cells for the CXCR4-tropic and CCR5-tropic virus and includes the the lytic and non-lytic immune responses. Computer-based simulations and a sensitivity analysis of the model predict that a persistent immune response suppresses the CXCR4-tropic virus to low levels and hence preventing a phenotypic switch. However, not only should the immune response be persistent, but it should have an efficient lytic immune response rather that an efficient non-lytic response. In addition, we also find that the availability of macrophage cells and enhanced viral kinetics are also crucial for the dominance of the R5 strain. We suggest that an altered host environment probably as a result of immune activation may explain the difference in coreceptor switching kinetics between HIV-1 subtype B and subtype C individuals. PMID- 17086374 TI - Outcomes of Lichtenstein hernioplasty for primary and recurrent inguinal hernia. AB - INTRODUCTION: The Lichtenstein hernioplasty for the repair of primary inguinal hernia in male patients is well established and constitutes the current gold standard. However a gold standard technique for the repair of recurrent inguinal hernia has not been established. The aim of this study was to analyze the outcomes of Lichtenstein hernioplasty for the repair of primary inguinal hernia and recurrent inguinal hernia, applying for that purpose the Qualitative and Quantitative Measurement Instrument (QQMI). METHODS: We studied 75 recurrent inguinal hernia patients and 287 primary inguinal hernia patients with a follow up period ranging from 60 to 107 months. RESULTS: The final QQMI score demonstrated that most patients in both groups reached scores between 8 and 11 points, with a significant difference in the maximum score (11 points) favoring primary hernia patients. CONCLUSIONS: All evaluated parameters showed better outcomes in primary hernia patients. Applying the QQMI, we have demonstrated that the outcomes of Lichtenstein hernioplasty are not similar between primary and recurrent inguinal hernia; there is a tendency toward better outcomes for primary inguinal hernia patients, although the Lichtenstein hernioplasty stands as a safe option for repair of recurrent inguinal hernias. PMID- 17086375 TI - The analysis of infection after polypropylene mesh repair of abdominal wall hernia. AB - AIM: The aim of the study was to evaluate the frequency of superficial and prosthetic mesh infection following polypropylene mesh repair of different abdominal wall hernia in individual patients and to analyze the manifestation, clinical process and outcomes in patients with prosthetic mesh infection. METHODS: This was a retrospective analysis of 375 patients with 423 implanted meshes for groin, femoral, umbilical, incisional and epigastric hernias, with a mean follow-up of 15 months (range: 3-73 months). RESULTS: The total superficial infection rate was 1.65%, and the rate of mesh infection was 0.94%. There were no statistically significant differences in prosthetic mesh infection between monofilament and multifilament meshes as well as between the different repair groups of hernias. The deep incisional surgical site infection after previous operation was established as a significant risk factor for prosthetic mesh infection in incisional hernia repair (P < 0.0001). Five cases of prosthetic mesh infection were presented and analyzed. CONCLUSIONS: There is no correlation between the superficial and prosthetic mesh infection. There may be difficulties in determining mesh infection and to choose the right tactic. The reconvalescence in all patients with mesh infection was achieved only after removal of the infected mesh. PMID- 17086376 TI - Liver tissue dissection: ultrasonic or RFA energy? AB - BACKGROUND: Hepatic resection is the only potential curative treatment for a wide variety of conditions. However, liver surgery is technically demanding and closely associated with a number of serious complications. New devices and techniques are currently being applied in practice, which will improve the surgical outcome. METHODS: This retrospective study compares two methods of liver parenchymal division: ultrasound energy, a modern but already widely used technique, and radio-frequency ablation (RFA), a completely novel method. The parameters investigated include the amount of blood transfused, the necessity of the Pringle maneuver, the length of time required for parenchymal division, and postoperative morbidity and mortality. The patients were divided into two groups. In one group (Group A), 15 patients underwent 17 ultrasound-assisted liver resections, in which ten metastatic tumors, six hepatomas, and one cholangiocarcinoma were resected. In Group B, 21 patients underwent 22 RFA assisted hepatectomies in which 11 metastatic tumors, ten hepatomas, and two cholangiocarcinomas were removed. RESULTS: Thirteen patients (87%) in Group A and 11 (52%) in Group B received a transfusion, with an average of 3.5 and 1.6 units of red blood cells, respectively. The Pringle maneuver was necessary in two cases in Group A but was unnecessary in Group B. The mean length of time required for parenchymal dissection was 124 min in Group A and 93.18 min in Group B. One (6.7%) and four (19%) complications were observed in Group A and B, respectively (statistically not significant). Mortality remained zero in both groups. CONCLUSIONS: RFA energy provides a novel reliable and safe alternative that can be used exclusively or as a supplement to the older techniques. Both resection time and amount of blood transfusion were reduced in the RFA group. PMID- 17086377 TI - Announcement of JPR Awards 2006. PMID- 17086378 TI - Indole-3-carbinol enhances ultraviolet B-induced apoptosis by sensitizing human melanoma cells. AB - Indole-3-carbinol (I3C) has been found to act against several types of cancer, while ultraviolet B (UVB) is known to induce the apoptosis of human melanoma cells. Here, we investigated whether I3C can sensitize G361 human melanoma cells to UVB-induced apoptosis. We examined the effects of combined I3C and UVB (I3C/UVB) at various dosages. I3C (200 microM)/UVB (50 mJ/cm(2)) synergistically reduced melanoma cell viability, whereas I3C (200 microM) or UVB (50 mJ/cm(2)), separately, had little effect on cell viability. DNA fragmentation assays indicated that I3C/UVB induced apoptosis. Further results show that I3C/UVB activates caspase-8, -3, and Bid and causes the cleavage of poly(ADP-ribose) polymerase. Moreover, I3C decreased the expression of the anti-apoptotic protein, Bcl-2, whereas UVB increased the translocation of Bax to mitochondria. Thus, an increased Bax/Bcl-2 ratio by I3C/UVB may result in melanoma apoptosis. In conclusion, our study demonstrated that I3C sensitizes human melanoma cells by down-regulating Bcl-2. PMID- 17086379 TI - Genetically engineered avidins and streptavidins. AB - Chicken avidin and bacterial streptavidin, (strept)avidin, are proteins widely utilized in a number of applications in life science, ranging from purification and labeling techniques to diagnostics, and from targeted drug delivery to nanotechnology. (Strept)avidin-biotin technology relies on the extremely tight and specific affinity between (strept)avidin and biotin (dissociation constant, K(d) approximately 10(-14)-10(-16) M). (Strept)avidins are also exceptionally stable proteins. To study their ligand binding and stability characteristics, the two proteins have been extensively modified both chemically and genetically. There are excellent accounts of this technology and chemically modified (strept)avidins, but no comprehensive reviews exist concerning genetically engineered (strept)avidins. To fill this gap, we here go through the genetically engineered (strept)avidins, summarizing how these constructs were designed and how they have improved our understanding of the structural and functional characteristics of these proteins, and the benefits they have provided for (strept)avidin-biotin technology. PMID- 17086380 TI - Betaine homocysteine S-methyltransferase: just a regulator of homocysteine metabolism? AB - Betaine homocysteine methyltransferase (BHMT), a Zn(2+)-dependent thiolmethyltransferase, contributes to the regulation of homocysteine levels, increases in which are considered a risk factor for cardiovascular diseases. Most plasma homocysteine is generated through the liver methionine cycle, in which BHMT metabolizes approximately 25% of this non-protein amino acid. This process allows recovery of one of the three methylation equivalents used in phosphatidylcholine synthesis through transmethylation, a major homocysteine producing pathway. Although BHMT has been known for over 40 years, the difficulties encountered in its isolation precluded detailed studies until very recently. Thus, the last 10 years, since the sequence became available, have yielded extensive structural and functional data. Moreover, recent findings offer clues for potential new functions for BHMT. The purpose of this review is to provide an integrated view of the knowledge available on BHMT, and to analyze its putative roles in other processes through interactions uncover to date. PMID- 17086381 TI - Regulation of nicotinic acetylcholine receptors by tyrosine kinases in the peripheral and central nervous system: same players, different roles. AB - Nicotinic acetylcholine receptors (nAChRs) exist in many subtypes and are found in the peripheral and central nervous system where they mediate or modulate synaptic transmission. We review how tyrosine phosphorylation and kinases regulate muscle and neuronal nAChRs. Interestingly, although some of the same kinase players interact with the various receptor subtypes, the functional consequences are different. While concerted action of MuSK, Abl- and Src-family kinases (SFKs) regulates the synaptic distribution of nAChRs at the neuromuscular junction, SFKs activate heteromeric neuronal nAChRs in adrenal chromaffin cells, thereby enhancing catecholamine secretion. In contrast, the activity of homomeric neuronal nAChRs, as found in the hippocampus, is negatively regulated by tyrosine phosphorylation and SFKs. It appears that tyrosine kinases provide the means to regulate all nAChRs; but the functional consequences, even those caused by the same kinase family, are specific for each receptor subtype and location. PMID- 17086382 TI - Therapeutic effect of pulsed electromagnetic field in conservative treatment of subacromial impingement syndrome. AB - Subacromial impingement syndrome (SIS) is a frequent cause of shoulder pain. Our purpose in this double-blinded, randomized, and controlled study was to demonstrate whether the pulsed electromagnetic field (PEMF) provides additional benefit when used with other conservative treatment modalities in acute phase rehabilitation program of SIS. Forty-six patients with unilateral shoulder pain who had been diagnosed as having SIS were included in this trial. The cases were randomly separated into two groups. All cases received a treatment program for 3 weeks consisting of Codman's pendulum exercises and subsequent cold pack gel application on shoulders with pain 5 times a day, restriction of daily activities that require the hands to be used over the head, and meloxicam tablet 15 mg daily. One group was given PEMF; the other group was given sham PEMF daily, 25 min per session, 5 days per week for 3 weeks. Shoulder pain during rest and activity and which causes disturbance of sleep was evaluated using a visual analogue scale, and total Constant score investigated shoulder function. Daily living activities were evaluated by shoulder disability questionnaire. Results were assessed before and after treatment. When compared with the baseline values, significant improvements in all these variables were observed at the end of the treatment in both groups (p<0.05). No significant difference between treatments was observed for any of these variables (p>0.05). There is no convincing evidence that electromagnetic therapy is of additional benefit in acute phase rehabilitation program of SIS. PMID- 17086383 TI - The effects of sulfasalazine treatment on enthesal abnormalities of inflammatory rheumatic diseases. AB - The aim of this study was to evaluate the effects of a 1-year course of sulfasalazine monotherapy on enthesal abnormalities of inflammatory rheumatic diseases (IRDs) using ultrasonography. Thirty-six patients with IRD including 20 patients with rheumatoid arthritis (RA) and 16 patients with ankylosing spondylitis (AS) (22 women, 14 men, mean ages 43.3 +/- 8.8 years), and 18 healthy controls (10 women, 8 men, mean ages 42.5 +/- 9.9 years) matched by age and body mass index were enrolled in this study. For the evaluation of enthesal structures, all patients and controls underwent ultrasonographic (USG) examinations of five enthesal sites of both lower limbs using high-resolution and Doppler USG. An ultrasonographic score of lower limb enthesitis was calculated using Glasgow ultrasound enthesitis scoring system (GUESS). Clinical and laboratory activities of IRD patients were also evaluated. Patient group was made to undergo 2 g/day sulfasalazine monotherapy for 1 year. All evaluations were made at the beginning of the treatment and repeated after 1 year follow-up. Results showed that the frequency of enthesal abnormalities of the IRD group was significantly higher than controls. On USG examination, 301/1,296 (23.2%) enthesal structures were abnormal in IRD patients, and 19/648 (2.93%) structures were abnormal in controls. Mean GUESS score of the IRD group (6.40 +/- 2.41) was also significantly higher than controls (1.79 +/- 1.60) (p < 0.001). Although there was a significant improvement in clinical and laboratory activity parameters of the IRD patients, significant decrease was not observed in enthesal abnormalities (295/1,296 enthesal structures-22.7%) and mean GUESS score (6.20 +/ 2.38) after 1 year sulfasalazine trial. Additionally, there was no significant improvement in enthesal abnormalities and mean GUESS scores of AS and RA subgroups separately. Sulfasalazine treatment was not found effective on enthesal abnormalities of IRD patients. Further studies with larger groups including other IRDs are required to validate our results. PMID- 17086384 TI - Ten years of clinical experience with adult onset Still's disease: is the outcome improving? AB - This study aims to report on the clinical and laboratory picture and the disease course and outcome in patients having adult onset Still's disease (AOSD), to briefly review existing literature on the subject, and to compare our findings with those previously reported. Results are reported for 28 patients with AOSD satisfying the preliminary criteria of Yamaguchi et al. seen in a teaching hospital over the last 10 years. A high percent of the patients with AOSD were women. The mean (+SD) age at disease onset was 27.8 (+8.4) years. We found fever in 100%, rash in 85%, arthritis in 64%, lymphadenopathy in 60%, splenomegaly in 57%, hepatomegaly in 35%, pleural effusion in 17.9%, and pericardial effusion in 3.6% of our patients. Leukocytosis was present in 96% of the patients, a normochromic, normocytic anemia in 54%, and an elevated erythrocyte sedimentation rate (ESR) in all. Serum ferritin levels were raised in 89% of the patients. The mean follow-up of the patients was 3.72 + 2.46 years. The mean delay in diagnosis was 7.32 + 18.0 months. The mean time to enter remission was 9.7 months. Self limited, intermittent, and chronic disease course was seen in 14.3, 57.1, and 28.6% of patients, respectively. The outcome was good in about 89% of patients, and mortality was nil. No particular clinical or laboratory variable was found to predict the subsequent disease course and outcome in our patients. On comparing our data with important previous series, we found a higher percentage of women and of patients presenting in the age group 16-35 years, a lower frequency of arthritis and pericardial effusion, and some other notable differences. Importantly, the disease course was benign, probably as an outcome of heightened awareness and less diagnostic delay than in the past, allowing for early, aggressive, and appropriate treatment. It is concluded that AOSD is now a relatively benign disease if diagnosed early and treated appropriately. PMID- 17086385 TI - Recent developments in methods and technology for analysis of biological samples by MALDI-TOF-MS. AB - Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) is widely used in a variety of fields because it has the characteristics of speed, ease of use, high sensitivity, and wide detectable mass range for obtaining molecular weights and for structural characterization of macromolecules. In this article we summarize recent developments in matrix additives, new matrices, and sample-pretreatment methods using off-probe or on probe techniques or nanomaterials for MALDI-TOF-MS analysis of biological samples. PMID- 17086386 TI - Development and characterisation of disposable gold electrodes, and their use for lead(II) analysis. AB - There is an increasing need to assess the harmful effects of heavy-metal-ion pollution on the environment. The ability to detect and measure toxic contaminants on site using simple, cost effective, and field-portable sensors is an important aspect of environmental protection and facilitating rapid decision making. A screen-printed gold sensor in a three-electrode configuration has been developed for analysis of lead(II) by square-wave stripping voltammetry (SWSV). The working electrode was fabricated with gold ink deposited by use of thick-film technology. Conditions affecting the lead stripping response were characterised and optimized. Experimental data indicated that chloride ions are important in lead deposition and subsequent analysis with this type of sensor. A linear concentration range of 10-50 microg L(-1) and 25-300 microg L(-1) with detection limits of 2 microg L(-1) and 5.8 microg L(-1) were obtained for lead(II) for measurement times of four and two minutes, respectively. The electrodes can be reused up to 20 times after cleaning with 0.5 mol L(-1) sulfuric acid. Interference of other metals with the response to lead were also examined to optimize the sensor response for analysis of environmental samples. The analytical utility of the sensor was demonstrated by applying the system to a variety of wastewater and soil sample extracts from polluted sites. The results are sufficient evidence of the feasibility of using these screen-printed gold electrodes for the determination of lead(II) in wastewater and soil extracts. For comparison purposes a mercury-film electrode and ICP-MS were used for validation. PMID- 17086387 TI - Estimation of diffusive boundary layer thickness in studies involving diffusive gradients in thin films (DGT). AB - Recent laboratory experiments and field investigations involving diffusive gradients in thin films (DGT) have shown that the thickness (delta) of the diffusive boundary layer (DBL), which can affect the accuracy of the technique, is generally not negligible. Accordingly, the determination of delta has become a matter of considerable practical importance. Though the problem has been addressed in the recent literature, there is room for some improvement. An expression for estimation of delta is presented here, and a practical procedure for determining delta and the concentration of DGT-labile species from sparse experimental data is proposed and illustrated by analyzing data from four experiments with DGT samplers of different diffusive gel thicknesses. PMID- 17086388 TI - Determination of glucosinolates in traditional Chinese herbs by high-performance liquid chromatography and electrospray ionization mass spectrometry. AB - A reversed-phase HPLC method has been developed for determination of twelve intact glucosinolates--glucoiberin, glucocheirolin, progoitrin, sinigrin, epiprogoitrin, glucoraphenin, sinalbin, gluconapin, glucosibarin, glucotropaeolin, glucoerucin, and gluconasturtiin--in ten traditional Chinese plants. The samples were extracted with methanol and the extracts were cleaned on an activated Florisil column. A mobile phase gradient prepared from methanol and 30 mmol L(-1) ammonium acetate at pH 5.0 enabled baseline separation of the glucosinolates. Glucosinolate detection was confirmed by quadrupole time-of flight tandem mass spectrometric analysis in negative-ionization mode. Detection limits ranged from 0.06 to 0.36 microg g(-1) when 5 g of dried plant was analyzed. Recoveries of the glucosinolates were better than 85% and precision (relative standard derivation, n = 3) ranged from 5.3 to 14.6%. Analysis of the glucosinolates provided scientific evidence enabling differentiation of three pairs of easily confused plants. PMID- 17086389 TI - SPR-based immunocapture approach to creating an interfacial sensing architecture: Mapping of the MRS18-2 binding site on retinoblastoma protein. AB - Biosensor technologies based on optical readout are widely used in protein protein interaction studies. Here we describe a fast and simple approach to the creation of oriented interfacial architectures for surface plasmon resonance (SPR) transducers, based on conventional biochemical procedures and custom reagents. The proposed protocol permits the oriented affinity-capture of GST fusion proteins by a specific antibody which is bound to protein A, which in turn has been immobilized on the transducer surface (after the surface has been modified by guanidine thiocyanate). The applicability of the method was demonstrated by studying the interaction between retinoblastoma tumor suppressor protein (pRb) and MRS18-2 proteins. The formation of the pRb-MRS18-2 protein complex was examined and the pRb binding site (A-box-spacer-B-box) was mapped. We have also shown that MRS18-2, which was detected as the Epstein-Barr virus encoded EBNA-6 binding partner using the yeast two-hybrid system, binds to pRb in GST pull-down assays. PMID- 17086390 TI - Fourier transform infrared (FT-IR) spectroscopy in bacteriology: towards a reference method for bacteria discrimination. AB - Rapid and reliable discrimination among clinically relevant pathogenic organisms is a crucial task in microbiology. Microorganism resistance to antimicrobial agents increases prevalence of infections. The possibility of Fourier transform infrared (FT-IR) spectroscopy to assess the overall molecular composition of microbial cells in a non-destructive manner is reflected in the specific spectral fingerprints highly typical for different microorganisms. With the objective of using FT-IR spectroscopy for discrimination between diverse microbial species and strains on a routine basis, a wide range of chemometrics techniques need to be applied. Still a major issue in using FT-IR for successful bacteria characterization is the method for spectra pre-processing. We analyzed different spectra pre-processing methods and their impact on the reduction of spectral variability and on the increase of robustness of chemometrics models. Different types of the Enterococcus faecium bacterial strain were classified according to chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis (PFGE). Samples were collected from human patients. Collected FT-IR spectra were used to verify if the same classification was obtained. In order to further optimize bacteria classification we investigated whether a selected combination of the most discriminative spectral regions could improve results. Two different variable selection methods (genetic algorithms (GAs) and bootstrapping) were investigated and their relative merit for bacteria classification is reported by comparing with results obtained using the entire spectra. Discriminant partial least-squares (Di-PLS) models based on corrected spectra showed improved predictive ability up to 40% when compared to equivalent models using the entire spectral range. The uncertainty in estimating scores was reduced by about 50% when compared to models with all wavelengths. Spectral ranges with relevant chemical information for Enterococcus faecium bacteria discrimination were outlined. PMID- 17086391 TI - Reduction of nonspecific protein binding on surface plasmon resonance biosensors. AB - Reduction of the nonspecific serum protein adsorption on a gold surface to levels low enough to allow the detection of biomarkers in complex media has been achieved using the N-hydroxysuccinimide (NHS) ester of 16-mercaptohexadecanoic acid. Carboxymethylated dextran (CM dextran), which is widely used, nonspecifically adsorbs enough proteins to mask the signal from target biomarkers in complex solutions such as serum or blood. The use of short-chain thiols greatly reduces the amount of nonspecific protein adsorption. Mixed layers of 11 mercaptoundecanoic acid or the NHS ester of 11-mercaptoundecanoic acid mixed layers with either 11-mercaptoundecanol or undecanethiol, and 16 mercaptohexadecanoic acid or the NHS ester of 16-mercaptohexadecanoic acid with hexadecanethiol, were also investigated for nonspecific protein binding properties as well as for biomarker signal response. The NHS ester of 16 mercaptohexadecanoic acid exhibits the largest signal for the biomarker myoglobin (including CM dextran) while offering a significantly diminished amount of nonspecific binding. The sensor has also been shown to detect interleukin-6 in cell culture media containing protein concentrations of at least 4 mg/mL. PMID- 17086394 TI - Jejunal Dieulafoy lesion with massive lower intestinal bleeding. PMID- 17086392 TI - The ability of the mesocortical dopamine system to operate in distinct temporal modes. AB - BACKGROUND: This review discusses evidence that cells in the mesocortical dopamine (DA) system influence information processing in target areas across three distinct temporal domains. DISCUSSIONS: Phasic bursting of midbrain DA neurons may provide temporally precise information about the mismatch between expected and actual rewards (prediction errors) that has been hypothesized to serve as a learning signal in efferent regions. However, because DA acts as a relatively slow modulator of cortical neurotransmission, it is unclear whether DA can indeed act to precisely transmit prediction errors to prefrontal cortex (PFC). In light of recent physiological and anatomical evidence, we propose that corelease of glutamate from DA and/or non-DA neurons in the VTA could serve to transmit this temporally precise signal. In contrast, DA acts in a protracted manner to provide spatially and temporally diffuse modulation of PFC pyramidal neurons and interneurons. This modulation occurs first via a relatively rapid depolarization of fast-spiking interneurons that acts on the order of seconds. This is followed by a more protracted modulation of a variety of other ionic currents on timescales of minutes to hours, which may bias the manner in which cortical networks process information. However, the prolonged actions of DA may be curtailed by counteracting influences, which likely include opposing actions at D1 and D2-like receptors that have been shown to be time- and concentration dependent. In this way, the mesocortical DA system optimizes the characteristics of glutamate, GABA, and DA neurotransmission both within the midbrain and cortex to communicate temporally precise information and to modulate network activity patterns on prolonged timescales. PMID- 17086395 TI - Increased presence of serotonin-producing cells in colons with diverticular disease may indicate involvement in the pathophysiology of the condition. AB - INTRODUCTION: Serotonin is an important neuroendocrine transmitter participating in the control of colonic motor activity through neural and biochemical mechanisms in the Enteric Nervous System (ENS). A possible pathophysiological factor for diverticular disease (DD) is altered colonic motility. The study compared the distribution of serotonin cells (SC) in the colonic mucosa of patients with diverticular disease to controls. METHODS: Sixteen paraffin specimens with sigmoid diverticular disease were selected and sections of bowel without diverticula from the same specimen were used as its own control. The resection margins from sixteen colonic specimens excised for sigmoid cancer were additional controls. Immunocytochemical staining for serotonin cells was performed on 4-mum tissue sections with polyclonal antibody (NCL-SEROTp). The number of serotonin-positive cells per ten microscopic fields (x200) was assessed in all groups and the staining distribution was defined as low (0-33%), moderate (>33-66%) and high (>66%) according to the percentage of the entire cell containing contrast material. The control specimens were blinded before analysis. Student's t test was used for statistical analysis and significance level was set as P < 0.05. RESULTS: The mean number of serotonin-positive cells per ten fields in the colonic mucosa of specimens with diverticular disease was significantly higher [252.44 (SD 90.64)] than the specimen's own control [147.31 (SD 50.16)] and at normal resection margins of cancer specimens [228.38 (SD 120.10)]. The paired analysis between diverticular disease specimens and its own control (paired t test) showed significant differences for moderate (P = 0.008), high (P = 0.001) and total (P = 0.002) number of serotonin cells. There was no evidence of significance between mean DD and cancer values. DISCUSSION: Increased presence of SCs and the higher proportion of high and moderate staining cells (indicating increased hormone content) indicate the possible role of serotonin in DD. This may be contributing to the pathogenesis of the condition by altered colonic motility in the affected segments in a similar way as in irritable bowel syndrome. PMID- 17086396 TI - Complications of endoscopic extraperitoneal radical prostatectomy (EERPE): prevention and management. AB - Endoscopic extraperitoneal radical prostatectomy (EERPE) is a further advancement of minimal invasive surgery as it overcomes the limitations of laparoscopic (transperitoneal) RPE by the strictly extraperitoneal route of access combining the advantages of minimal invasive surgery with the advantages of an extraperitoneal procedure. Endoscopic extraperitoneal radical prostatectomy has many advantages but is not without complications. The aim of this review article is to describe the most common complications of this procedure. Complications associated with endoscopic extraperitoneal radical prostatectomy are: vascular injury--bleeding--haematoma, bowel injury, lymphocele, injury to the bladder- ureter, port site hernia, anastomotic leakage--stricture, obturator nerve injury- paralysis, gas embolism, catheter blockage, and miscellaneous like perineal pain, pubic osteitis, infection- urosepsis. The present review paper focuses on the identification and management of these complications The incidence of most complications directly correlates with the surgeons' experience, and the various complications are related to technical errors rather than to the technique itself. The laparoscopist performing endoscopic/ laparoscopic radical prostatectomy should be aware of all these complications. He should be able to recognise promptly, treat efficiently, and ideally prevent these complications. PMID- 17086397 TI - Genes for alkaline/neutral invertase in rice: alkaline/neutral invertases are located in plant mitochondria and also in plastids. AB - Two cDNA clones (OsNIN1 and OsNIN3) encoding an alkaline/neutral invertase localized in organelles were identified from rice. The deduced amino acid sequences of these cDNA clones showed high homology to other plant alkaline/neutral invertases. Semi-quantitative reverse transcription polymerase chain reaction revealed that the expression of OsNIN1 was constitutive and independent of organ difference, although its expression level was low. Analyses using five types of web software for the prediction of protein localization in the cell, Predotar, PSORT, Mitoprot, TargetP, and ChloroP, strongly supported the possibility that OsNIN1 is transported into the mitochondria and that OsNIN3 is transported into plastids. Transient expression of fusion proteins combining the amino terminal region of these two proteins with sGFP demonstrated that N OsNIN1::GFP and N-OsNIN3::GFP fusion proteins were transported into the mitochondria and plastids, respectively. We expressed the OsNIN1 protein in vitro and revealed that the translated protein had an invertase activity. These results clearly indicate that some of alkaline/neutral invertases are located in plant organelles, mitochondria and plastids, and that they might have a novel physiological function in plant organelles. PMID- 17086398 TI - Osmotin induces cold protection in olive trees by affecting programmed cell death and cytoskeleton organization. AB - Osmotin is a pathogenesis-related protein exhibiting cryoprotective functions. Our aim was to understand whether it is involved in the cold acclimation of the olive tree (Olea europaea L.), a frost-sensitive species lacking dormancy. We exposed olive trees expressing tobacco osmotin gene under the 35S promoter (35S:osm) [in the same manner as wild type (wt) plants] to cold shocks in the presence/absence of cold acclimation, and monitored changes in programmed cell death (PCD), cytoskeleton, and calcium ([Ca2+]c) signalling. In the wt, osmotin was immunolocalized only in cold-acclimated plants, and in the tissues showing PCD. In the 35S:osm clones, the protein was detected also in the non-acclimated plants, and always in the tissues exhibiting PCD. In the non-acclimated wt protoplasts exposed to cold shock, a transient decrease in phallotoxin signal suggests a temporary disassembly of F-actin, a transient increase occurred instead in 35S:osm protoplasts exposed to the same shock. Transient increases in [Ca2+]c were observed only in the wt protoplasts. However, when F-actin was depolymerized by cytochalasin or latrunculin, and microtubules by colchicine, increase in [Ca2+]c also occurred in the 35S:osm protoplasts. Successive cold shocks caused transient rises in [Ca2+]c and transient decreases in the phallotoxin signal in wt protoplasts. No change occurred in [Ca2+]c occurred in the 35S:osm protoplasts. The phallotoxin signal transiently increased at the first shock, but did not change after the subsequent shocks, and an overall signal reduction occurred with shock repetition. Following acclimation, no cold shock-induced change in [Ca2+]c levels and F-actin signal occurred either in wt or 35S:osm protoplasts. The results show that osmotin is positively involved in the acclimation-related PCD, in blocking the cold-induced calcium signalling, and in affecting cytoskeleton in response to cold stimuli. PMID- 17086399 TI - Cadmium-induced alterations of the structural features of pectins in flax hypocotyl. AB - In the course of our studies on the putative role of pectins in the control of cell growth, we have investigated the effect of cadmium on their composition, remodelling and distribution within the epidermis and fibre tissues of flax hypocotyl (Linum usitatissimum L.). Cadmium-stressed seedlings showed a significant inhibition of growth whereas the hypocotyl volume did not significantly change, due to the swelling of most tissues. The structural alterations consisted of significant increase of the thickness of all cell walls and the marked collapse of the sub-epidermal layer. The pectic epitopes recognized by the anti-PGA/RGI and JIM5 antibodies increased in the outer parts of the epidermis (external tangential wall and junctions) and fibres (primary wall and junctions). Concomitantly, there was a remarkable decrease of JIM7 antibody labelling and consequently an increase of the ratio JIM5/JIM7. Conversely, the ratio JIM7/JIM5 increased in the wall domains closest to the plasmalemma, which would expel the cadmium ions from the cytoplasm. The hydrolysis of cell walls revealed a cadmium-induced increase of uronic acid in the pectic matrix. Sequential extractions showed a remodelling of both homogalacturonan and rhamnogalacturonan I. In fractions enriched in primary walls, the main part of the pectins became cross-linked and could be extracted only with alkali. In fractions enriched in secondary walls, the homogalacturonan moieties were found more abundantly in the calcium-chelator extract while the rhamnogacturonan level increased in the boiling water extract. PMID- 17086400 TI - Investigation of ferulate deposition in endosperm cell walls of mature and developing wheat grains by using a polyclonal antibody. AB - A polyclonal antibody has been raised against ferulic acid ester linked to arabinoxylans (AX). 5-O-feruloyl-alpha-L-arabinofuranosyl(1-->4)-beta-D xylopyranosyl was obtained by chemical synthesis, and was coupled to bovine serum albumin for the immunization of rabbit. The polyclonal antibody designated 5-O Fer-Ara was highly specific for 5-O-(trans-feruloyl)-L-arabinose (5-O-Fer-Ara) structure that is a structural feature of cell wall AX of plants belonging to the family of Gramineae. The antibody has been used to study the location and deposition of feruloylated AX in walls of aleurone and starchy endosperm of wheat grain. 5-O-Fer-Ara began to accumulate early in aleurone cell wall development (beginning of grain filling, 13 days after anthesis, DAA) and continued to accumulate until the aleurone cells were firmly fixed between the starchy endosperm and the nucelus epidermis (19 DAA). From 26 DAA to maturity, the aleurone cell walls changed little in appearance. The concentration of 5-O-Fer Ara is high in both peri- and anticlinal aleurone cell walls with the highest accumulation of 5-O-Fer-Ara at the cell junctions at the seed coat interface. The situation is quite different in the starchy endosperm: whatever the stage of development, a low amount of 5-O-Fer-Ara epitope was detected. Contrary to what was observed for aleurone cell walls, no peak of accumulation of feruloylated AX was noticed between 13 and 19 DAA. Visualization of labelled Golgi vesicles suggested that the feruloylation of AX is intracellular. The distribution of (5-O Fer-Ara) epitope is further discussed in relation to the role of ferulic acid and its dehydrodimers in cell wall structure and tissue organization of wheat grain. PMID- 17086401 TI - Expression of Arabidopsis phytochelatin synthase in Indian mustard (Brassica juncea) plants enhances tolerance for Cd and Zn. AB - Phytochelatins (PCs) are heavy metal binding peptides that play an important role in sequestration and detoxification of heavy metals in plants. In this study, our goal was to develop transgenic plants with increased tolerance for and accumulation of heavy metals from soil by expressing an Arabidopsis thaliana AtPCS1 gene, encoding phytochelatin synthase (PCS), in Indian mustard (Brassica juncea L.). A 35S promoter fused to a FLAG-tagged AtPCS1 cDNA was expressed in Indian mustard, and transgenic lines, designated pc lines, were evaluated for tolerance to and accumulation of Cd and Zn. Transgenic plants with moderate AtPCS1 expression levels showed significantly higher tolerance to Cd and Zn stress, but accumulated significantly less Cd and Zn than wild type plants in both shoot and root tissues. However, transgenic plants with highest expression of the transgene did not exhibit enhanced Cd and Zn tolerance. Shoots of Cd treated pc plants had significantly higher levels of phytochelatins and thiols than wild-type plants. Significantly lower concentrations of gluthatione in Cd treated shoot and root tissues of transgenic plants were observed. Moderate expression levels of phytochelatin synthase improved the ability of Indian mustard to tolerate certain levels of heavy metals, but at the same time did not increase the accumulation potential for Cd and Zn. PMID- 17086402 TI - Underexpression of the plant NOTCHLESS gene, encoding a WD-repeat protein, causes pleitropic phenotype during plant development. AB - WD-repeat proteins are involved in a breadth of cellular processes. While the WD repeat protein encoding gene NOTCHLESS has been involved in the regulation of the Notch signaling pathway in Drosophila, its yeast homolog Rsa4p was shown to participate in 60S ribosomal subunit biogenesis. The plant homolog ScNLE was previously characterized in Solanum chacoense (ScNLE) as being involved in seed development. However, expression data and reduced size of ScNLE underexpressing plants suggested in addition a role during shoot development. We here report the detailed phenotypic characterization of ScNLE underexpressing plants during shoot development. ScNLE was shown to be expressed in actively dividing cells of the shoot apex. Consistent with this, ScNLE underexpression caused pleiotropic defects such as a reduction in aerial organ size, a reduction in some organ numbers, delayed flowering, and an increase in stomatal index. Analysis of adaxial epidermal cells revealed that both cell number and cell size were reduced in mature leaves of ScNLE underexpressing lines. Two-hybrid screens with the Nle domain and the WD-repeat domain of ScNLE allowed the isolation of homologs of yeast MIDASIN and NSA2 genes, the products of which are involved in 60S ribosomal subunit biogenesis in yeast. A ScNLE-GFP chimeric protein was localized in both the cytoplasm and nucleus. These data altogether suggest that ScNLE likely plays a role in 60S ribosomal subunit biogenesis, which is essential for proper cellular growth and proliferation during plant development. PMID- 17086403 TI - Chronic kidney disease in children: state of the art. PMID- 17086404 TI - Teaching articles: a new goal of the journal. PMID- 17086405 TI - A prospective study on the use of mycophenolate mofetil in children with cyclosporine-dependent nephrotic syndrome. AB - Cyclosporine A (CsA) has relieved children with steroid-dependent nephrotic syndrome (NS) from steroid toxicity. However, most patients frequently relapse again when CsA is withdrawn, resulting in the development of CsA nephropathy for its long-term use. In order to assess the efficacy of mycophenolate mofetil (MMF) therapy, we prospectively analyzed 12 children with idiopathic steroid-dependent NS requiring long-term CsA therapy with MMF for at least 6 months. Mean follow-up after starting MMF was 11 months (range 6-42). The mean MMF dose required was 610+/-95 mg/m(2)/12 h, which maintained mean predose mycophenolic acid (C0-MPA) levels of 2.4+/-1.1 mcg/ml. Treatment with MMF resulted in CsA and/or prednisolone (PSL) sparing, with a reduction in mean CsA dose from 3.5+/-1.3 to 1.5+/-2.4 mg/kg/day (p<0.01), and mean PSL dose from 0.29+/-0.16 to 0.21+/-0.11 mg/kg/day (p<0.05). Nine of 12 patients (75%) were finally able to be weaned off CsA. Mean relapse rates decreased from 2.7+/-1.6 to 0.6+/-0.9 episodes/year (p<0.01). Relapse-free ratio on MMF therapy was lower in patients whose average C0-MPA levels were less than 2 mcg/ml (p<0.05). Our experience demonstrates that MMF therapy results in significant CsA and/or steroid sparing and reduction in relapse rates in children with CsA-dependent NS. PMID- 17086406 TI - Utilization of glutamate/creatine ratios for proton spectroscopic diagnosis of meningiomas. AB - INTRODUCTION: Our purpose was to determine the potential of metabolites other than alanine to diagnose intracranial meningiomas on proton magnetic resonance spectroscopy (MRS). METHODS: Using a 1.5-T MR system the lesions were initially identified on FLAIR, and T1- and T2-weighted images. Employing standard point resolved spectroscopy (PRESS) for single voxel proton MRS (TR 1500 ms, TE 30 ms, 128 acquisitions, voxel size 2 x 2 x 2 cm, acquisition time 3.12 min), MR spectra were obtained from 5 patients with meningiomas, from 20 with other intracranial lesions, and from 4 normal controls. Peak heights of nine resonances, including lipid, lactate, alanine, NAA (N-acetylaspartate), beta/gamma-Glx (glutamate + glutamine), creatine, choline, myo-inositol, and alpha-Glx/glutathione, were measured in all spectra. The relative quantity of each metabolite was measured as the ratio of its peak height to the peak height of creatine. RESULTS: Relative quantities of alpha-Glx/glutathione, beta/gamma-Glx, and total Glx/glutathione were significantly elevated in meningiomas compared to the 20 other intracranial lesions and the normal control brains. Alanine was found in four of five meningiomas, but lactate partially masked the alanine in three meningiomas. None of the other lesions or control brains showed an alanine peak. The one meningioma with no alanine and the three others with lactate had elevated Glx. CONCLUSION: While alanine is a relatively unique marker for meningioma, our results support the hypothesis that the combination of glutamate/creatine ratios and alanine on proton MRS is more specific and reliable for the diagnosis of meningiomas than alanine alone. PMID- 17086407 TI - Implant detectibility of intervertebral disc spacers in post fusion MRI: evaluation of the MRI scan quality by using a scoring system--an in vitro study. AB - INTRODUCTION: Intervertebral spacers for anterior spine fusion are made of different materials, such as titanium and cobalt chromium alloys and carbon fiber reinforced polymers. Implant-related susceptibility artifacts can decrease the quality of MRI scans. The aim of this cadaveric study was to demonstrate the extent that implant-related MRI artifacting affects the postfusion differentiation of determined regions of interest (ROIs). METHODS: In six cadaveric porcine spines, we evaluated the postimplantation MRI scans of a titanium, cobalt-chromium and carbon spacer that differed in shape and surface qualities. A spacer made of human cortical bone was used as a control. A defined evaluation unit was divided into ROIs to characterize the spinal canal as well as the intervertebral disc space. Considering 15 different MRI sequences read independently by an interobserver-validated team of specialists the artifact affected image quality of the median MRI slice was rated on a score of 0-3. A maximum score of 18 points (100%) for the determined ROIs was possible. RESULTS: Turbo spin echo sequences produced the best scores for all spacers and the control. Only the control achieved a score of 100%. For the determined ROI maximum scores for the cobalt-chromium, titanium and carbon spacers were 24%, 32% and 84%, respectively. CONCLUSION: By using favored T1 TSE sequences the carbon spacer showed a clear advantage in postfusion spinal imaging. Independent of artifact dimensions, the scoring system used allowed us to create an implant related ranking of MRI scan quality in reference to the bone control. PMID- 17086408 TI - Modified buddy wire technique for coil embolization of posterior circulation aneurysms. AB - INTRODUCTION: In coil embolization of a posterior circulation aneurysm, tortuosity or a small vessel caliber frequently prevents placement of a guiding catheter in the vertebral artery. Even in such difficult situations, however, coil embolization is the only treatment option when the patient's clinical condition is very poor, or the aneurysm is inaccessible by microsurgery. METHODS: Three patients with a posterior circulation aneurysm were treated by coil embolization with the use of a modified buddy wire technique to stabilize a guiding catheter that could not be placed into the vertebral artery. RESULTS: The guiding catheter was stabilized in the subclavian artery and the microcatheter was firmly supported. As a result, coil embolization was performed safely in all patients. There were no procedural complications. CONCLUSION: The modified buddy wire technique could provide patients with a chance to undergo endovascular surgery despite having a tortuous or narrow vertebral artery. PMID- 17086409 TI - Endovascular treatment of the vertebral artery origin in patients with symptoms of vertebrobasilar ischemia. AB - INTRODUCTION: We report our experience with the endovascular treatment of the vertebral artery origin in patients presenting with symptomatic vertebrobasilar ischemia and compare our results with those reported in the literature. METHODS: In 25 patients, 28 procedures were performed. Patients presented with posterior circulation ischemic symptoms despite optimal medical therapy with antiplatelet drugs and had a digital subtraction angiogram demonstrating stenosis of the origin of the vertebral artery greater than 50%. Retrospective review of the medical records, clinical notes and radiologic-procedural reports was performed. RESULTS: Of the 25 patients, 18 were male and 7 female. Their ages ranged from 50 to 84 years. In 23 of the 25 patients the contralateral vertebral artery was occluded, hypoplastic, absent, or had greater than 50% stenosis. In 13 of the 25 patients angiographic evidence of significant anterior circulation disease was demonstrated. In 18 of the 25 patients the left vertebral artery was affected. The mean stenosis was 82.6%. Follow-up records were available in 19 patients. The mean follow-up was 24 months. Five of the 19 patients had recurrent symptoms of vertebrobasilar ischemia and three patients were retreated. Of the 28 procedures performed, 23 were angioplasty/stenting and 5 were angioplasties alone. Overall technical success was achieved in 26 of the 28 procedures (92.8%). No procedure related transient ischemic attack, stroke or death was noted. CONCLUSION: Endovascular treatment of the vertebral artery origin in this patient population is feasible, safe, and effective. There are some questions regarding the long term follow-up and rate of restenosis and clinical recurrences that are yet to be answered. PMID- 17086410 TI - Comparative study on central metabolic fluxes of Bacillus megaterium strains in continuous culture using 13C labelled substrates. AB - Fluxes of central carbon metabolism [glycolysis, pentose phosphate pathway (PPP), tricarboxylic acid cycle (TCA cycle), biomass formation] were determined for several Bacillus megaterium strains (DSM319, WH320, WH323, MS941) in C- and N limited chemostat cultures by (13)C labelling experiments. The labelling patterns of proteinogenic amino acids were analysed by GC/MS and therefrom flux ratios at important nodes within the metabolic network could be calculated. On the basis of a stoichiometric metabolic model flux distributions were estimated for the different B. megaterium strains used at various cultivation conditions. Generally all strains exhibited similar metabolic flux distributions, however, several significant changes were found in (1) the glucose flux entering the PPP via the oxidative branch, (2) the reversibilities within the PPP, (3) the relative fluxes of pyruvate and acetyl-CoA fed to the TCA cycle, (4) the fluxes around the pyruvate node involving a futile cycle. PMID- 17086411 TI - Trehalose synthesis in Saccharomycopsis fibuligera does not respond to stress treatments. AB - Synthesis of trehalose in Saccharomycopsis fibuligera sdu under various stress conditions was investigated. Neither the activation of trehalose-6-phosphate synthase (SfTPS1) nor the change in trehalose content was observed under stress exposure of S. fibuligera sdu cells. The results of reverse transcription polymerase chain reaction, which was performed with the specific primers designed to target the SfTPS1 gene fragment cloned from this strain, also showed that all stress treatments did not increase the expression of SfTPS1 gene. These results demonstrated that synthesis of trehalose in response to stress conditions in S. fibuligera sdu clearly differs from that of Saccharomyces cerevisiae and most other fungi. The phylogenetic analysis of the amino acid sequence deduced from the SfTPS1 gene fragment showed that the SfTPS1 sequence formed a separate family that was far related to S. cerevisiae TPS1. The yeast strain, which can accumulate a large amount of trehalose under normal growth conditions, has many applications and TPS1 gene in such strain may have unique use in transgenic organisms. PMID- 17086412 TI - Optimization of critical medium components using response surface methodology for biomass and extracellular polysaccharide production by Agaricus blazei. AB - Response surface methodology (RSM) was applied to optimize the critical medium ingredients of Agaricus blazei. A three-level Box-Behnken factorial design was employed to determine the maximum biomass and extracellular polysaccharide (EPS) yields at optimum levels for glucose, yeast extract (YE), and peptone. A mathematical model was then developed to show the effect of each medium composition and its interactions on the production of mycelial biomass and EPS. The model predicted the maximum biomass yield of 10.86 g/l that appeared at glucose, YE, peptone of 26.3, 6.84, and 6.62 g/l, respectively, while a maximum EPS yield of 348.4 mg/l appeared at glucose, YE, peptone of 28.4, 4.96, 5.60 g/l, respectively. These predicted values were also verified by validation experiments. The excellent correlation between predicted and measured values of each model justifies the validity of both the response models. The results of bioreactor fermentation also show that the optimized culture medium enhanced both biomass (13.91 +/- 0.71 g/l) and EPS (363 +/- 4.1 mg/l) production by Agaricus blazei in a large-scale fermentation process. PMID- 17086413 TI - Decolorization of a dye industry effluent by Aspergillus fumigatus XC6. AB - The strain Aspergillus fumigatus XC6 isolated from mildewing rice straw was evaluated for its ability to decolorize a dye industry effluent. The strain was capable of decolorizing dyes effluent over a pH range 3.0-8.0 with the dyes as sole carbon and nitrogen sources. The optimum pH was 3.0; however, supplemented with either appropriate nitrogen sources (0.2% NH(4)Cl or (NH(4))(2)SO(4) ) or carbon sources (1.0% sucrose or potato starch), the strain decolorized the effluent completely at the original pH of the dyes effluent. Therefore, A. fumigatus XC6 is an efficient strain for the decolorization of reactive textile dyes effluents, and it might be a practical alternative in dyeing wastewater treatment. PMID- 17086414 TI - Identification of genetic markers to distinguish the virulent and avirulent subspecies of Pantoea stewartii by comparative proteomics and genetic analysis. AB - Pantoea stewartii subsp. stewartii (Pnss), the causal agent of Stewart's bacterial wilt and leaf blight of maize and sweet corn, is one of the quarantine pathogens in many countries and regions. In contrast, P. stewartii subsp. indologenes (Pnsi), the closely related subspecies of Pnss, is avirulent on these plants. In this study, the protein expression profiles of these two subspecies were compared using two-dimensional gel electrophoresis analysis. Twenty-one unique protein spots consistently detected in Pnss but not in Pnsi were analyzed by mass spectrometry. Some of these Pnss-specific proteins are known to be essential for virulence and survival in host, such as FoxR and HrcJ, which are the key components of iron uptake and Type III secretion systems, respectively. For further genetic analysis, six Pnss-specific proteins were characterized by peptide sequencing. Southern and Northern blot analyses revealed that the differences in protein expression profiles of the two subspecies were either due to the discrepancy at genome level or because of the variations in transcriptional expression. The results provide novel genetic markers to distinguish the two closely related subspecies and may also serve as useful clues for investigation of the genetic basis accounting for their sharp difference in virulence. PMID- 17086415 TI - Down-regulation of lactate dehydrogenase-A by siRNAs for reduced lactic acid formation of Chinese hamster ovary cells producing thrombopoietin. AB - Lactate, one of the major waste products in mammalian cell culture, can inhibit cell growth and affect cellular metabolism at high concentrations. To reduce lactate formation, lactate dehydrogenase-A (LDH-A), an enzyme catalyzing the conversion of glucose-derived pyruvate to lactate, was down-regulated by an expression vector of small interfering RNAs (siRNA) in recombinant Chinese hamster ovary (rCHO) cells producing human thrombopoietin (hTPO). Three clones expressing low levels of LDH-A, determined by reverse transcription-PCR and an enzyme activity test, were established in addition to a negative control cell line. LDH-A activities in the three clones were decreased by 75-89%, compared with that of the control CHO cell line, demonstrating that the effect of siRNA is more significant than that of other traditional methods such as homologous recombination (30%) and antisense mRNA (29%). The specific glucose consumption rates of the three clones were reduced to 54-87% when compared to the control cell line. Similarly, the specific lactate production rates were reduced to 45 79% of the control cell line level. In addition, reduction of LDH-A did not impair either cell proliferation or hTPO productivity. Taken together, these results show that the lactate formation rate in rCHO cell culture can be efficiently reduced through the down-regulation of LDH via siRNA. PMID- 17086416 TI - [Primary central nervous system degeneration in elderly patients. Characteristic imaging features]. AB - Despite further development of new magnetic resonance imaging techniques, e.g., diffusion tensor imaging and 1H magnetic resonance spectroscopy, structural imaging will continue to play a major role in the diagnosis of primary central nervous system degeneration in ageing. Characteristic imaging patterns of multisystem atrophies and primary dementias as well as differential diagnostic features are demonstrated. While such features may have high specificity, their sensitivity is low especially in cross-sectional studies. Longitudinal studies are the optimal method to characterize the dynamic neuroanatomical correlates of the disease. However, according to disease duration and progression, neuroimaging will show increased overlapping and convergence of pathological changes in multisystem atrophy as well as in dementia. PMID- 17086417 TI - Probing structural changes in the alpha and beta domains of copper- and silver substituted metallothionein by emission spectroscopy and electrospray ionization mass spectrometry. AB - Steady-state emission spectra, excited-state lifetimes, kinetic data, and mass spectroscopic properties are reported for Ag(I)- and mixed Ag(I)/Cu(I) substituted alpha and beta domains of recombinant human metallothionein (MT1a). Kinetic analysis of the changes in the Cu(I) emission spectra during the stepwise displacement of Cu(I) ions by Ag(I) at room temperature shows that the rate of displacement of Cu(I) is unexpectedly slow. Although the first Ag(I) added results in major changes in the Cu(I)-MT binding site, Cu(I) displacement by Ag(I) does not take place until the addition of the third Ag(I), and is completed by the addition of the seventh Ag(I). The emission from Ag(I) and mixed Cu(I)/Ag(I)-MT species at 77 K shows that the band maxima shift as a function of Ag(I) loading, which can be correlated with shifts in coordination geometry from trigonal to digonal. Two phosphorescence lifetimes were detected for the Ag(I) substituted alpha and beta domains of MT, which are attributed to the presence of Ag(I) ions in two different environments. The lifetime of Ag(I)-substituted MT was found to be shorter when the Ag(I)-MT species were formed by Ag(I) additions to the Cu(I)-substituted alpha and beta fragments than when the Ag(I)-MT species were formed from the apo-alpha and apo-beta fragments, suggesting the formation of structurally different Ag(I)-MT clusters. Electrospray ionization mass spectrometric studies suggest the metallation reactions of Ag(I) with MT take place in a series of steps to form a series of Ag(I)-substituted MT species. Ag(I)-substituted MT species are not detected until past the addition of 3 mol equiv of Ag(I), suggesting that cluster formation begins only at this point, stabilizing the metallated species sufficiently to survive ionization. PMID- 17086418 TI - Preconception care--when and what: the attitude of Israeli gynaecologists to preconception counseling. AB - OBJECTIVES: Preconception care is a form of preventive care and its implementation might improve general women's health as well as improving pregnancy outcome. Our aim was to survey the attitudes of Israeli gynecologists regarding preconception counseling. METHODS: E-mailed questionnaires were sent to gynecologists in an Israeli gynecology network. The questionnaire included two identical sets of questions; one was regarding the care of reproductive aged women who visit their gynecologist for various reasons and the second was regarding the care of women in their first prenatal visit. Answers were scored (from 3-always to 0-never), summed and compared using Chi-square and paired t tests. RESULTS: Mean score for the preconception set was significantly lower than the prenatal set 11.3 (62.7%) versus 16.9 (93.7%) respectively, P < 0.001. Folic acid supplementation and genetic screening tests were recommended to most women in 99 and 94% of first prenatal care visit compared to only 42 and 62% of women who were not pregnant (P < 0.001). CONCLUSIONS: Gynecologists fail to recognize proper opportunities for preconception care. Increased awareness and concrete guidelines concerning timing and content of preconception counseling might be helpful. PMID- 17086419 TI - Paraoxonase-1 activity as a marker of atherosclerosis is not associated with low bone mineral density in healthy postmenopausal women. AB - The aging process is associated with an increasing prevalence of osteoporosis and atherosclerosis, but it is uncertain if these two conditions are interrelated. Serum paraoxonase-1 (PON1) is a high-density lipoprotein (HDL) associated enzyme that has been implicated in the pathogenesis of atherosclerosis. Our aims of the study were to investigate (1) serum paraoxonase and arylesterase activities and, lipid hydroperoxide (LOOH) levels in healthy postmenopausal women and (2) whether there were any associations between these enzyme activities and bone mineral density (BMD). A total of 97 generally healthy postmenopausal women were enrolled in the study. BMD was measured at lumbar spine (LS) and femoral neck (FN) with dual energy X-ray absorptiometry. Serum paraoxonase and arylesterase activities were measured spectrophotometrically. LOOH levels were measured by iodometric assay. In this population, 50 (51%) women had BMD T scores < -2.5 at the LS and/or FN defined as osteoporosis and 47 (49%) of them had normal BMDs. Serum paraoxonase, arylesterase, and LOOH activities were not significantly different between osteoporotic and nonosteoporotic postmenopausal women. There were also no correlations between paraoxonase, arylesterase, LOOH activities, and LS BMD and FN BMD. We conclude that there may be not good evidence to support a direct relationship between osteoporosis and atherosclerosis in these subjects. However, prospective studies with larger groups are needed to investigate this issue further. PMID- 17086420 TI - Genes up-regulated during red coloration in UV-B irradiated lettuce leaves. AB - Molecular analysis of gene expression differences between green and red lettuce leaves was performed using the SSH method. BlastX comparisons of subtractive expressed sequence tags (ESTs) indicated that 7.6% of clones encoded enzymes involved in secondary metabolism. Such clones had a particularly high abundance of flavonoid-metabolism proteins (6.5%). Following SSH, 566 clones were rescreened for differential gene expression using dot-blot hybridization. Of these, 53 were found to overexpressed during red coloration. The up-regulated expression of six genes was confirmed by Northern blot analyses. The expression of chalcone synthase (CHS), flavanone 3-hydroxylase (F3H), and dihydroflavonol 4 reductase (DFR) genes showed a positive correlation with anthocyanin accumulation in UV-B-irradiated lettuce leaves; flavonoid 3',5'-hydroxylase (F3',5'H) and anthocyanidin synthase (ANS) were expressed continuously in both samples. These results indicated that the genes CHS, F3H, and DFR coincided with increases in anthocyanin accumulation during the red coloration of lettuce leaves. This study show a relationship between red coloration and the expression of up-regulated genes in lettuce. The subtractive cDNA library and EST database described in this study represent a valuable resource for further research for secondary metabolism in the vegetable crops. PMID- 17086421 TI - Cytogenetic evidence for diversity of two nuclei within a single diplomonad cell of Giardia. AB - Giardia intestinalis is an ancient protist that causes the most commonly reported human diarrheal disease of parasitic origin worldwide. An intriguing feature of the Giardia cell is the presence of two morphologically similar nuclei, generally considered equivalent, in spite of the fact that their karyotypes are unknown. We found that within a single cell, the two nuclei differ both in the number and the size of chromosomes and that representatives of two major genetic groups of G. intestinalis possess different karyotypes. Odd chromosome numbers indicate aneuploidy of Giardia nuclei, and their stable occurrence is suggestive of a long term asexuality. A semi-open type of Giardia mitosis excludes a chromosome interfusion between the nuclei. Differences in karyotype and DNA content, and cell cycle-dependent asynchrony are indicative of diversity of the two Giardia nuclei. PMID- 17086422 TI - The distinct effects of three tandem repeats of C3d in the immune responses against tumor-associated antigen hCGbeta by DNA immunization. AB - Several examples have shown that C3d3, when fused to a corresponding antigen, had a strong adjuvant effect on certain specific antibody production. In a previous study, we attempted to prove that this was the case of the human chorionic gonadotrophin beta chain (hCGbeta)-induced immunity following DNA vaccination. However, we found that C3d3 when fused to hCGbeta inhibited rather than enhanced the antigen-specific immune response. In the present study, using hCGbeta DNA vaccine preparations, we demonstrated that C3d3 inhibited the antigen-specific humoral antibody response and several other immune responses, such as the hCGbeta specific lymphoproliferation. Such inhibitory effects of C3d3 were not related to the expression level of the target protein, the gender of the test mice, or the vector used. Contrastingly, C3d3 fused with the envelope protein of hepatitis B virus (PreS2/S) used as a control system resulted in the enhancement of both humoral and cell-mediated antigen-specific immune responses against HBV-preS2/S, which was consistent with other groups' adjuvant-effect findings. We further showed that the mechanisms involved in the inhibitory effect of C3d3 might be possible due to impairing the function of antigen presenting B lymphocytes and reducing the expression of transcription factors (T-bet and GATA-3) and cytokine IL-4. Collectively, unlike its usual expected adjuvant function, the fusion of C3d3 with the tumor-associated antigen hCGbeta was found to inhibit both humoral and cell-mediated antigen-specific immune responses. These findings indicate that research concerning tumor immune escapes and vaccine designs require further extensive attention. PMID- 17086423 TI - Vascular endothelial growth factor inhibits the function of human mature dendritic cells mediated by VEGF receptor-2. AB - Dendritic cells (DCs) are the most potent antigen-presenting cells and play a central role in the host-antitumor immunity. Since it has been reported that vascular endothelial growth factor (VEGF) inhibits the functional maturation of immature-DCs and impairs DC differentiation, it is important to elucidate the mechanisms of VEGF-induced DC-dysfunction. To investigate the effects of VEGF against human mature DCs, we investigated how VEGF affects mature DCs with regards to phenotype, induction of apoptosis, IL-12(p70) production and the antigen-presenting function evaluated by allogeneic mixed leukocyte reaction (allo-MLR). We generated monocyte-derived DCs matured with lipopolysaccharide, OK 432 or pro-inflammatory cytokine cocktails. As a result, VEGF treatment did not alter the mature DCs with regard to phenotype, IL-12(p70) production and induction of apoptosis. As a novel and important finding, VEGF inhibited the ability of mature DCs to stimulate allogeneic T cells. Furthermore, this VEGF induced DC dysfunction was mainly mediated by VEGF receptor-2 (VEGF R2). These observations were confirmed by the findings that the VEGF-induced DC dysfunction was recovered by anti-human VEGF neutralizing mAb or anti-human VEGF R2 blocking mAb, and that placenta growth factor (PlGF), VEGF R1-specific ligand, did not have any effect against mature DCs. Some modalities aiming at reversing mature-DC dysfunction induced by VEGF will be needed in order to induce the effective antitumor immunity. PMID- 17086424 TI - Fluvastatin reduced liver injury in rat model of extrahepatic cholestasis. AB - Inhibitors of 3-hydroxy-3methylglutarly coenzyme A, reductase, namely statins, exert pleiotropic actions beyond lipid-lowering effects. In ex vivo and in vitro studies, statins have antioxidative and antiinflammatory effects. Herein, we sought to determine whether treatment with fluvastatin (FV) would be beneficial in a rat model of common bile duct ligation (BDL)-induced liver injury. Female rats were subjected to a sham (n=10) or BDL (n=20). Obstructive jaundice was induced in rats by the ligation and division of the common bile duct. Three days after operation, rats subjected to CBDL were randomized to receive treatment with either FV (10 mg/kg) or saline every day over a 10 days experimental period. High levels of alanine aminotransferase, aspartate aminotransferase, and gamma glutamyltransferase decreased significantly (P<0.05) in animals treated with FV with compared to saline-administrated BDL animals. Compared with sham-operated rats, CBDL rats showed significantly higher levels of total nitrite and nitrate, malondihaldehyde, tumor necrosis factor alpha, myeloperoxidase, and lower concentrations of glutathione, superoxide dismutase, and catalase in the liver tissue (P<0.001). All of these changes were significantly attenuated (P<0.05) by treatment with FV after CBDL. CBDL was associated with increased apoptosis and nuclear factor kappa beta expression in saline-treated rats. Treatment with FV also decreased these parameters. These data support the view that FV ameliorates hepatic inflammation, lipid peroxidation, and tissue injury in rats subjected to CDBL. FV warrants further evaluation as an adjunctive treatment to ameliorate liver injury from extrahepatic biliary obstruction. PMID- 17086425 TI - Typhoid intestinal perforation in children: a continuing scourge in a developing country. AB - Typhoid intestinal perforation is a principal cause of morbidity and mortality affecting both adults and children. This study aims to evaluate the pattern of typhoid intestinal perforation and outcome of its management in Nigerian children. The records of consecutive children managed for typhoid intestinal perforation at Juth, between 1996 and 2005 have been reviewed. There were 184 children (M:F = 1.04:1), with a mean of 5.8 years (range 4 to 15 years). More than a half (62.5%) of the patients were in the 5-6 year age group. The incidence peaked in April-May, and November-December. Increased incidence of typhoid perforation was observed between 2002 and 2005. All patients presented with the classic features of typhoid enteric perforation. Hypokalaemia and anaemia were common at presentation. Only 75 (40.6%) patients had operation within 24 h of perforation. The types of surgery included simple excision of the edges of the peroration and closure (74.5%), wedge resection and closure in (14.5%), segmental resection with primary end-to-end anastomosis (3.6%) and right hemi-colectomy with ileo-colic or ileo-transverse anastomosis. Wound infection and dehiscence, anastomotic breakdown with faecal fistula, intra peritoneal abscesses and chest infections were the main post operative complications. The overall mortality rate was 22.8 (42). Excision and simple closure was associated with the least incidence of anastomotic breakdown and operation time. The incidence of typhoid intestinal perforation is on the increase at our institution. Early limited surgery provides optimal results. PMID- 17086426 TI - Conservation and divergence of candidate class B genes in Akebia trifoliata (Lardizabalaceae). AB - There is evidence that gene duplication and diversification within the MADS-box gene family had significant impact on floral architecture. In this study, we report the isolation of four class B homologous genes from Akebia trifoliata, termed AktAP3_1, AktAP3_2, AktAP3_3, and AktPI. Phylogenetic analysis indicates that the three AktAP3 paralogs were produced by two gene duplication events and AktAP3_2 and AktAP3_3 are recent paralogs, which are yielded by the duplication before the origin of the genus Akebia. In situ hybridization demonstrates that these genes are mainly expressed in the stamens and carpels of A. trifoliata, but in differential patterns, similar to those in other basal eudicot and basal angiosperm species. AktAP3_3 and AktPI are expressed in the developing petaloid perianth, suggesting that the petaloidy of the perianth is caused by the expression of class B genes. Reverse transcriptase polymerase chain reaction analyses indicate that these genes are expressed in both male and female flowers, but at different levels. We explore the interaction behavior of the class B proteins in the basal eudicots using yeast two-hybrid system for the first time. The AktAP3_1/2/3 proteins and the AktPI protein can form obligate heterodimers, but at different strength. From the mRNA expression and protein interaction patterns of the duplicated copies of the AktAP3 genes, we conclude that subfunctionalization very likely contributes to the maintenance of multiple AP3 like gene copies in A. trifoliata. PMID- 17086427 TI - Spatial orientation in the bushcricket Leptophyes punctatissima (Phaneropterinae; Orthoptera): I. Phonotaxis to elevated and depressed sound sources. AB - Many species of acoustically interacting insects live in a complex, arboreal or semi-arboreal habitat. Thus mate finding by phonotaxis requires sound localization in the horizontal and vertical plane. Here we investigated the ability of the duetting bushcricket Leptophyes punctatissima to orient to one of three speakers, positioned at different levels in an artificial grid system, where each point in space could be reached by the male with almost equal probability. The system was designed analogous to a spherical calotte model of bismuth, where, once the male arrived at any nodal point had to decide between only three directions: either up or down and/or left and right. This design does not favour any phonotactic path of the males. All 12 males tested reached the three speaker positions (one in the horizontal plane, one elevated by 45 degrees , one depressed by 45 degrees relative to the starting position) with only little deviation from the shortest possible path. There was no significant difference with respect to the whole phonotactic time needed, the number of segments passed, or the number of stimuli received for the different speaker positions. This remarkable spatial orientation is achieved although the insects have no specialized external ear structures such as mammals, or some owls. PMID- 17086428 TI - Meniscus suture. PMID- 17086429 TI - Preoperative bone quality as a factor in dual-energy X-ray absorptiometry analysis comparing bone remodelling between two implant types. AB - Recently it was shown that the design changes from the ABG-I to ABG-II hip stem resulted in a better, although not significant, proximal bone preservation. Our hypothesis was that by matching patients for preoperative bone quality, statistical power would increase and that the trend of better proximal bone preservation in ABG-II might become significant. Twenty-four ABG-II patients were compared to two different ABG-I groups: (1) 25 patients from our earlier prospective study and (2) a group of 24 patients selected to perfectly match the ABG-II group regarding gender, age and preoperative bone quality. Postoperative changes in periprosthetic bone mineral density (BMD) were quantified at 2 years postoperatively using DEXA scanning. Bone preservation (less BMD loss) was better for the ABG-II than the ABG-I (all two groups) in the proximal zones 1 and 7. In Gruen zone 7, a statistically significant difference was found for group B (p = 0.03). By matching patients for preoperative bone quality and gender, a statistical significant difference was found in proximal bone preservation in favour of ABG-II. In future comparative bone remodelling studies using DEXA, patients should be matched for preoperative bone quality and gender. PMID- 17086430 TI - Streptococcus agalactiae CAMP factor/protein B does not bind to human IgG. AB - CAMP factor is an extracellular cytolytic protein produced by Streptococcus agalactiae. CAMP factor has been reported to bind the Fc fragments of immunoglobulin G (IgG) and has therefore also been called protein B, in analogy to protein A of Staphylococcus aureus. We attempted to characterize the interaction of protein B with IgG in more detail. In contrast to protein A, CAMP factor does not inhibit the activation of complement by hemolysin antibodies bound to sheep red cell surfaces. IgG also failed to inhibit the co-hemolytic activity of CAMP factor, which is in disagreement with previous findings. After co-incubation, CAMP factor and IgG were cleanly separated by gel filtration, indicating that no binding had occurred. PMID- 17086431 TI - Nasal mucociliary clearance in total laryngectomized patients. AB - We measured the nasal mucociliary activity in total laryngectomy patients, and to compare among themselves, smokers, diabetics and normal population besides that to investigate the short and long term effects of total laryngectomy on nasal mucosa. The study includes 39 patients who had total laryngectomy between the January 1998-August 2005 and 36 volunteer healthy individuals. To examine the early and late changes on nasal mucosa the patients that had total laryngectomy separated into two groups as operated before August 2003 (> 2 years), as operated after August 2003 (< 2 years). Moreover diabetic patients and smokers are differently grouped. This study is performed in Haydarpasa Numune Hospital for Research and Education, Department of Otorhinolaryngology, between May 2005 September 2005. Mucociliary clearance measurement is performed by saccharin test for all patients and control group. The mucociliary clearance of the patients (diabetics-nondiabetics, smokers-non smokers) are measured and examined if they show any difference among themselves. For the patients operated between the dates August 2003-2005 (< 2 years group) the nasal mucociliary clearance time average is found 8,15 +/- 2.06 min., for the patients operated before August 2003 (> 2 years group) the mean time is found 23.79 +/- 5.58 min., for the control group the mean time is found 14.5 +/- 3.55 min In operated group the patients who are diabetics has longer mucociliary clearance time than nondiabetics, and the difference is statistically significant. Similarly in control group diabetics has longer mucociliary clearance time than nondiabetics. As another parameter smoking, comparing the groups among themselves, smokers have longer mucocilliary clearance time than nonsmokers. In total laryngectomy patients hypersecratory phase is produced in early period and nasal mucosal clearance is increased. On the other hand, dependent on chronic infections nasal mucosa is atrophied and nasal mucosal clearance is disrupted. However smoking and diabetes mellitus also damage the mucocilliary clearance. PMID- 17086434 TI - Medication-taking behavior in a cohort of patients with inflammatory bowel disease. AB - Recent studies have shown a low adherence rate to maintenance treatment in patients with inflammatory bowel disease (IBD). We sought to assess the medication-taking behavior in a cohort of patients with IBD. We prospectively included IBD patients from the outpatient clinic who agreed to answer a questionnaire about prescribed treatment and adherence. Physicians registered clinical data including prescribed medications. Two hundred fourteen patients (115 Crohn's disease/99 ulcerative colitis) were included. The most prescribed medications were oral mesalazine (56.5%) and immunomodulators (41.1%). Forty three percent of patients admitted to occasionally forgetting to take their medication but only 7.5% of them did it voluntary. Oral mesalazine and azathioprine were the drugs with the poorest compliance, with nonadherence rates of 45% and 25% of the total prescribed doses, respectively. The only factor associated with a better adherence was a more complicated course of the disease steroid dependency, steroid refractoriness, need for infliximab treatment, hospitalization, or surgery (P=.02). Twenty percent of patients admitted to self medicating. An important proportion of patients with IBD admit to forget some doses of the prescribed medication in the setting of a specialized unit of a referral centre. PMID- 17086435 TI - Sparse cerebellar innervation can morph the dynamics of a model oculomotor neural integrator. AB - The oculomotor integrator is a brainstem neural network that converts velocity signals into the position commands necessary for eye-movement control. The cerebellum can independently adjust the amplitude of eye-movement commands and the temporal characteristics of neural integration, but the percentage of integrator neurons that receive cerebellar input is very small. Adaptive dynamic systems models, configured using the genetic algorithm, show how sparse cerebellar inputs could morph the dynamics of the oculomotor integrator and independently adjust its overall response amplitude and time course. Dynamic morphing involves an interplay of opposites, in which some model Purkinje cells exert positive feedback on the network, while others exert negative feedback. Positive feedback can be increased to prolong the integrator time course at virtually any level of negative feedback. The more these two influences oppose each other, the larger become the response amplitudes of the individual units and of the overall integrator network. PMID- 17086436 TI - Images in neuro-oncology: a case of POEMS (Polyneuropathy, Organomegaly, Endocrinopathy, Monoclonal protein and Skin changes) in a patient with multicentric Castleman's disease. PMID- 17086437 TI - Migrant Latino day laborers and intentions to test for HIV. AB - This cross sectional study utilized convenience sampling to investigate the HIV testing intentions of 290 sexually active, male, migrant Latino day laborers, 18 years old or older. The findings indicate that day laborers are indeed at risk for HIV. Nearly two-thirds of the men intended to test for HIV in the next year. Men who were at higher risk of infection from their sexual encounters with female partners were more likely to intend to test, as were men making less money and men who did not perceive themselves to be at risk. Men who attended high school or beyond, and reported using a condom with casual female partners, were much more likely to intend to test than men who did not attend school and reported no such condom use. The prevalence of high risk sex and the low rates of condom use have implications for the increased transmission of HIV. PMID- 17086438 TI - The basic structure of community early intervention programs for children with autism: provider descriptions. AB - Autism researchers have identified a set of common effective practice elements for early intervention (EI) (e.g., intensive programming). The current study examined the reported use of common elements of effective interventions in community EI settings. Eighty EI providers reported about their programs. The majority of participants reported using common effective elements, however, the depth and quality of the use of these elements was highly variable. Taking community program structure into account in future research will facilitate the development of methodologies, which immediately fit into the context of community programming rather than requiring program adaptation for use in the real world. Recommendations for using current community program structure to improve use of evidence-based practices are discussed. PMID- 17086439 TI - On intersubjective engagement in autism: a controlled study of nonverbal aspects of conversation. AB - Does autism involve a deficit in intersubjective engagement with other persons? We studied nonverbal communication in children and adolescents with and without autism (n = 12 per group), group-matched for chronological age and verbal mental age, during 3 min of a videotaped interview. In keeping with previous studies, there were only subtle but potentially revealing group differences on behavioral ratings. Participants with autism made fewer head-shakes/nods (but not smiles) when the interviewer was talking, and the interviewer made fewer head-shakes/nods when participants were talking. Yet there were marked group differences on reliable 'subjective' ratings of (a) affective engagement and (b) the smoothness of reciprocal interaction. We interpret the findings in terms of a group difference in identification between conversational partners. PMID- 17086440 TI - Processing of affective speech prosody is impaired in Asperger syndrome. AB - Many people with the diagnosis of Asperger syndrome (AS) show poorly developed skills in understanding emotional messages. The present study addressed discrimination of speech prosody in children with AS at neurophysiological level. Detection of affective prosody was investigated in one-word utterances as indexed by the N1 and the mismatch negativity (MMN) of auditory event-related potentials (ERPs). Data from fourteen boys with AS were compared with those for thirteen typically developed boys. These results suggest atypical neural responses to affective prosody in children with AS and their fathers, especially over the RH, and that this impairment can already be seen at low-level information processes. Our results provide evidence for familial patterns of abnormal auditory brain reactions to prosodic features of speech. PMID- 17086441 TI - A comparative study of the use and understanding of self-presentational display rules in children with high functioning autism and Asperger's disorder. AB - The use and understanding of self-presentational display rules (SPDRs) was investigated in 21 children with high-functioning autism (HFA), 18 children with Asperger's disorder (AspD) and 20 typically developing (TD) children (all male, aged 4- to 11-years, matched on mental age). Their behaviour was coded during a deception scenario to assess use of SPDRs; understanding of SPDRs was assessed via three real/apparent emotion-understanding vignettes. The children with HFA and AspD used less effective SPDRs than the TD children, but there were no group differences in understanding SPDRs. The children with HFA and AspD did not differ on their use or understanding of SPDRs, and the results are discussed in relation to the similarities and differences between these diagnostic conditions. PMID- 17086442 TI - Paraquat-induced oxidative stress in drosophila melanogaster: effects of melatonin, glutathione, serotonin, minocycline, lipoic acid and ascorbic acid. AB - The efficacy of melatonin, glutathione, serotonin, minocycline, lipoic acid and ascorbic acid in counteracting the toxicity of paraquat in Drosophila melanogaster was examined. Male Oregon wild strain flies were fed for 5 days with control food or food containing the test substance. They were transferred in groups of five to vials containing only filter paper soaked with 20 mM paraquat in 5% sucrose solution. Survival was determined 24 and 48 h later. All the substances assayed increased the survival of D. melanogaster. At equimolar concentrations (0.43 mM) melatonin was more effective than serotonin, lipoic acid and ascorbic acid. However, lower concentrations of glutathione (0.22 mM) and minocycline (0.05 mM) were as efficient as melatonin. The highest survival rate (38.6%) after 48 h of paraquat treatment was found with 2.15 mM of lipoic acid. No synergistic effect of melatonin with glutathione, serotonin, minocycline, lipoic acid and ascorbic acid was detected. PMID- 17086443 TI - Induced expression of cathepsins and cystatin C in a murine model of demyelination. AB - While proteolytic enzymes are involved in the pathogenesis of multiple sclerosis (MS), the involvement of cathepsins has not been characterized in detail. To better understand the role of cathepsins, cDNA microarray analysis was used to study the brains of proteolipid protein transgenic (plp ( tg ) /-) mice, an animal model that closely mimics the failure of remyelination in MS. Analysis revealed upregulated expression of cathepsins L, H and B and their inhibitor, cystatin C. By in situ hybridization, the induction of cathepsins was primarily limited to microglia/macrophages of the white matter, with continuous expression from 2 to 8 months of age. Elevated protein level of cathepsins was confirmed at 4 months of age. In contrast, elevated expression of cystatin C was found in astrocytes. The ratio of microglia/macrophages to astrocytes increased throughout the course of demyelination, suggesting that the ratio of secreted cathepsins to cystatin C increased during that period. We propose that in MS, remyelination may be impaired by increasing activity of cathepsins inadequately controlled by cystatin C. PMID- 17086444 TI - The nucleolar protein SURF-6 is essential for viability in mouse NIH/3T3 cells. AB - SURF-6 is a bona fide nucleolar protein comprising an evolutionary conserved family that extends from human to yeast. The expression of the mammalian SURF-6 has been recently found to be regulated during the cell cycle. In order to determine the importance of SURF-6 in mammalian cells, we applied the Tet-On system to regulate conditionally, in response to tetracycline, the expression of an antisense RNA (asRNA) that targets Surf-6 mRNA in mouse NIH/3T3 cells. Induced Surf-6 asRNA caused an effective depletion of SURF-6 protein resulted in cell death and in an apparent arrest in the G1 phase of the cell cycle. These results provide for the first time evidence that expression of SURF-6 is essential for mammalian cell viability, and suggest that SURF-6 might participate in the progression of cell cycle. PMID- 17086445 TI - Retrograde regulation of nuclear gene expression in CW-CMS of rice. AB - The CW-cytoplasmic male sterility (CMS) line has the cytoplasm of Oryza rufipogon Griff, and mature pollen is morphologically normal under an optical microscope but lacks the ability to germinate; restorer gene Rf17 has been identified as restoring this ability. The difference between nuclear gene expression in mature anthers was compared for the CW-CMS line, [cms-CW] rf17rf17, and a maintainer line with normal cytoplasm of Oryza sativa L., [normal] rf17rf17. Using a 22-k rice oligoarray we detected 58 genes that were up-regulated more than threefold in the CW-CMS line. Expression in other organs was further investigated for 20 genes using RT-PCR. Five genes, including genes for alternative oxidase, were found to be preferentially expressed in [cms-CW] rf17rf17 but not in [normal] rf17rf17 or [cms-CW] Rf17Rf17. Such [cms-CW] rf17rf17-specific gene expression was only observed in mature anthers but not in leaves, stems, or roots, indicating the presence of anther-specific mitochondrial retrograde regulation of nuclear gene expression, and that Rf17 has a role in restoring the ectopic gene expression. We also used a proteomic approach to discover the retrograde regulated proteins and identified six proteins that were accumulated differently. These results reveal organ-specific induced mitochondrial retrograde pathways affecting nuclear gene expression possibly related to CMS. PMID- 17086446 TI - Prevalence of urinary incontinence in community dwelling men: a cross sectional nationwide epidemiological survey. AB - PURPOSE: Epidemiological studies on urinary incontinence (UI) in adult men of all ages are scarce. We aim to describe the UI phenomenon among community dwelling adult males in the United States (US). MATERIALS AND METHODS: We analyzed data from male respondents to a 14-item mailed UI symptoms questionnaire to the National Family Opinion (NFO) World Group Panel of 45,000 households matching the US 2000 census population on five key demographic elements. RESULTS: A total of 29,903 households responded, of which 21,590 were male heads of household (mean age 50 +/-15.2 years old). A total of 2,059 men (12.7%) reported symptoms of UI during the last 30 days. Symptoms of urge urinary incontinence (UUI), stress urinary incontinence (SUI), mixed urinary incontinence (MUI) symptoms, and other types of urinary incontinence (OUI) symptoms, were reported by 44.6%, 24.5%, 18.8%, and 12.3%, respectively. Among those with UI symptoms, the proportion of UUI and MUI symptoms increases with age while SUI symptoms decrease as age increases. Of those with UI symptoms, 44% consulted a physician of whom 57% did so within 6 months of the onset of symptoms. Treatments reported included absorbent pads (48%), prescription medication (30%), surgical procedure (18%), and intermittent catheterization (4%). CONCLUSIONS: Almost 1 in 10 males reported symptoms of UI. The UI prevalence rate increases with age and UUI symptoms is the most prevalent type reported. Less than half of men with UI symptoms sought professional help and half of them did so within 6 months of onset of UI symptoms. PMID- 17086447 TI - Hemodynamic response to exercise as measured by the solar IKG impedance cardiography module and correlation with metabolic variables. AB - OBJECTIVE: Impedance Cardiography (ICG) has been shown to be a feasible and accurate method for non-invasive measurement of cardiac index (CI). Aim of this investigation was the correlation of hemodynamic variables under exercise as measured by a specific ICG-monitor (Solar IKG-Modul, Version 3.0, GE-Healthcare, Freiburg, Germany) with metabolic variables. METHODS: Ten healthy volunteers were included in the investigation doing ergometer exercise (5 min equilibration followed by 5 min each at 50, 75, 100 and 125 W). Hemodynamic parameters were obtained by ICG. Metabolic variables were assessed by indirect calorimetry with the Deltatrac II Metabolic monitor using a helmet system for spontaneous respiration. RESULTS: CI increased throughout exercise (baseline: 3.0 +/- 0.4 l/min/m(2); 125 W: 4.8 +/- 0.5 l/min/m(2)). Heart rate (baseline: 87.2 +/- 13.4 bpm; 125 W: 152.7 +/- 22.4 bpm) and contractility (velocity index) (baseline: 48.9 +/- 9.3/1000 s; 125 W: 70.5 +/- 10.0/1000 s) showed a continuous rise while the stroke index decreased after an initial rise (baseline: 35.0 +/- 4.6 ml/m(2); 50 W: 37.6 +/- 4.9 ml/m(2); 75 W: 41.2 +/- 5.9 ml/m(2); 125 W: 32.3 +/- 6.1 ml/m(2)). VO(2) (baseline: 335.2 +/- 84.1 ml/min; 125 W: 1298.9 +/- 282.3 ml/min) and VCO(2)(baseline: 255.4 +/- 74.5 ml/min; 125 W: 1342.5 +/- 282.5 ml/min) increased throughout exercise. There was a good correlation in the individual fits between hemodynamic and metabolic variables. CONCLUSION: CI in healthy volunteers, as measured by the Solar IKG-Modul, correlates well with O(2) consumption and CO(2)-production in individual subjects, thus indicating the metabolic needs under exercise conditions in healthy individuals. PMID- 17086448 TI - Conductivity-based hematocrit measurement during cardiopulmonary bypass. AB - OBJECTIVE: In a recent clinical study on the reliability of a point-of-care (POC) analyzer, we described a downward bias in hematocrit measurement during cardiopulmonary bypass leading potentially to overtreatment with packed red cells. We hypothesized that the detected deviation is caused by variations in electrolyte concentration rather than by colloids used. METHODS: Blood was sampled from patients before cardiac surgery to obtain undiluted anticoagulated whole blood samples (n = 53). From each sample, four dilution series covering a hematocrit range of 15-30% were made using NaCl (0.9%), modified gelatine (4%), hydroxyethylstarch (6%), or a potassium-based (16 mEq/l) solution, respectively. In each dilution series, hematocrit was measured by POC and via the "golden standard" microcentrifugal method to determine whether the deviation of the POC analyzer to the microcentrifuge was dependent on the type and dilution level of the solution used. RESULTS: In contrast to the colloid-based dilution series, the crystalloids revealed a significant downward bias of the POC-analyzer with respect to the microcentrifuge (p < 0.05). Due to the correction algorithm for sodium in the POC-analyzer, this deviation was nearly constant for NaCl (mean of difference: -1.8 +/- 0.1%), but increased significantly in case of the potassium based solution (up to -8.2 +/- 0.4% after 1.5-times dilution). The starch- and gelatine-based solutions led to a significant upward bias (p < 0.05) that increased with progressing dilution (up to 1.2 +/- 0.1% for hydroxyethylstarch and up to 1.3 +/- 0.1% for modified gelatine after 1.5-times dilution). CONCLUSIONS: Conductivity-based POC hematocrit measurement suffers from biases due to changes of the plasma constituents. The downward bias in hematocrit as often seen during cardiopulmonary bypass is driven by changes of different electrolyte concentration rather than by colloids used per se. PMID- 17086449 TI - Response of arsenic-induced oxidative stress, DNA damage, and metal imbalance to combined administration of DMSA and monoisoamyl-DMSA during chronic arsenic poisoning in rats. AB - Arsenic and its compounds cause adverse health effects in humans. Current treatment employs administration of thiol chelators, such as meso-2,3 dimercaptosuccinic acid (DMSA) and sodium 2,3-dimercaptopropane 1-sulfonate (DMPS), which facilitate its excretion from the body. However, these chelating agents are compromised by number of limitations due to their lipophobic nature, particularly in case of chronic poisoning. Combination therapy is a new approach to ensure enhanced removal of metal from the body, reduced doses of potentially toxic chelators, and no redistribution of metal from one organ to another, following chronic metal exposure. The present study attempts to investigate dose related effects of two thiol chelators, DMSA and one of its new analogues, monoisoamyl dimercaptosuccinic acid (MiADMSA), when administered in combination with the aim of achieving normalization of altered biochemical parameters suggestive of oxidative stress and depletion of inorganic arsenic following chronic arsenic exposure. Twenty-five adult male Wistar rats were given 25 ppm arsenic for 10 weeks followed by chelation therapy with the above chelating agents at a dose of 0.3 mmol/kg (orally) when administered individually or 0.15 mmol/kg and 0.3 mmol/kg (once daily for 5 consecutive days), respectively, when administered in combination. Arsenic exposure led to the inhibition of blood delta-aminolevulinic acid dehydratase (ALAD) activity and depletion of glutathione (GSH) level. These changes were accompanied by significant depletion of hemoglobin, RBC and Hct as well as blood superoxide dismutase (SOD) acitivity. There was an increase in hepatic and renal levels of thiobarbituric acid-reactive substances, while GSH:GSSG ratio decreased significantly, accompanied by a significant increase in metallothionein (MT) in hepatocytes. DNA damage based on denaturing polyacrylamide gel electrophoresis revealed significant loss in the integrity of DNA extracted from the liver of arsenic-exposed rats compared to that of normal animals. These changes were accompanied by a significant elevation in blood and soft-tissue arsenic concentration. Co-administration of DMSA and MiADMSA at lower dose (0.15 mmol/kg) was most effective not only in reducing arsenic-induced oxidative stress but also in depleting arsenic from blood and soft tissues compared to other treatments. This combination was also able to repair DNA damage caused following arsenic exposure. We thus recommend combined administration of DMSA and MiADMSA for achieving optimum effects of chelation therapy. PMID- 17086450 TI - Identification of three distinct phylogenetic groups of CAX cation/proton antiporters. AB - Ca(2+)/cation antiporter (CaCA) proteins are integral membrane proteins that transport Ca(2+) or other cations using the H(+) or Na(+) gradient generated by primary transporters. The CAX (for CAtion eXchanger) family is one of the five families that make up the CaCA superfamily. CAX genes have been found in bacteria, Dictyostelium, fungi, plants, and lower vertebrates, but only a small number of CAXs have been functionally characterized. In this study, we explored the diversity of CAXs and their phylogenetic relationships. The results demonstrate that there are three major types of CAXs: type I (CAXs similar to Arabidopsis thaliana CAX1, found in plants, fungi, and bacteria), type II (CAXs with a long N-terminus hydrophilic region, found in fungi, Dictyostelium, and lower vertebrates), and type III (CAXs similar to Escherichia coli ChaA, found in bacteria). Some CAXs were found to have secondary structures that are different from the canonical six transmembrane (TM) domains-acidic motif-five TM domain structure. Our phylogenetic tree indicated no evidence to support the cyanobacterial origin of plant CAXs or the classification of Arabidopsis exchangers CAX7 to CAX11. For the first time, these results clearly define the CAX exchanger family and its subtypes in phylogenetic terms. The surprising diversity of CAXs demonstrates their potential range of biochemical properties and physiologic relevance. PMID- 17086451 TI - The frequency of eubacterium-to-eukaryote lateral gene transfers shows significant cross-taxa variation within amoebozoa. AB - Single-celled bacterivorous eukaryotes offer excellent test cases for evaluation of the frequency of prey-to-predator lateral gene transfer (LGT). Here we use analysis of expressed sequence tag (EST) data sets to quantify the extent of LGT from eubacteria to two amoebae, Acanthamoeba castellanii and Hartmannella vermiformis. Stringent screening for LGT proceeded in several steps intended to enrich for authentic events while at the same time minimizing the incidence of false positives due to factors such as limitations in database coverage and ancient paralogy. The results were compared with data obtained when the same methodology was applied to EST libraries from a number of other eukaryotic taxa. Significant differences in the extent of apparent eubacterium-to-eukaryote LGT were found between taxa. Our results indicate that there may be substantial inter taxon variation in the number of LGT events that become fixed even between amoebozoan species that have similar feeding modalities. PMID- 17086452 TI - Alternative starts of transcription, several paralogues, and almost-fixed interspecific differences of the gene fruitless in a hemimetabolous insect. AB - In fruit flies, the gene fruitless (fru) governs the establishment of the potential for male sexual behavior. We partially cloned fru from a hemimetabolous insect for the first time and we compared fru among three closely related and acoustically communicating grasshopper species: Chorthippus biguttulus, C. brunneus, and C. mollis. The fru of grasshoppers is organized similarly to fru of holometabolous insects, with a BTB and Zn-finger domains separated by a nonconserved repetitive linker. As in Drosophila, several transcripts of fru are found in grasshoppers. We also present evidence for the coexistence of several copies of fru in the grasshopper genome. Within species these copies are almost identical and carry almost-fixed species-specific differences. This suggests that the paralogous copies of fru in grasshoppers do not evolve independently from each other. PMID- 17086453 TI - Mitogenomic evolution and interrelationships of the Cypriniformes (Actinopterygii: Ostariophysi): the first evidence toward resolution of higher level relationships of the world's largest freshwater fish clade based on 59 whole mitogenome sequences. AB - Fishes of the order Cypriniformes are almost completely restricted to freshwater bodies and number > 3400 species placed in 5 families, each with poorly defined subfamilies and/or tribes. The present study represents the first attempt toward resolution of the higher-level relationships of the world's largest freshwater fish clade based on whole mitochondrial (mt) genome sequences from 53 cypriniforms (including 46 newly determined sequences) plus 6 outgroups. Unambiguously aligned, concatenated mt genome sequences (14,563 bp) were divided into 5 partitions (first, second, and third codon positions of the protein-coding genes, rRNA genes, and tRNA genes), and partitioned Bayesian analyses were conducted, with protein-coding genes being treated in 3 different manners (all positions included; third codon positions converted into purine [R] and pyrimidine [Y] [RY-coding]; third codon positions excluded). The resultant phylogenies strongly supported monophyly of the Cypriniformes as well as that of the families Cyprinidae, Catostomidae, and a clade comprising Balitoridae + Cobitidae, with the 2 latter loach families being reciprocally paraphyletic. Although all of the data sets yielded nearly identical tree topologies with regard to the shallower relationships, deeper relationships among the 4 major clades (the above 3 major clades plus Gyrinocheilidae, represented by a single species Gyrinocheilus aymonieri in this study), were incongruent depending on the data sets. Treatment of the rapidly saturated third codon-position transitions appeared to be a source of such incongruities, and we advocate that RY-coding, which takes only transversions into account, effectively removes this likely "noise" from the data set and avoids the apparent lack of signal by retaining all available positions in the data set. PMID- 17086454 TI - Inadvertent fracture of a plastic biliary stent during a combined percutaneous endoscopic procedure: a word of caution regarding self-locking pigtail biliary catheters. PMID- 17086455 TI - Endovascular treatment of a renal artery branch aneurysm. AB - A 58-year-old woman was admitted to our institution because of a left renal artery branch saccular aneurysm with a 2 cm diameter. Due to a hostile abdomen and the infrarenal location, an endovascular approach was chosen. A Jostent Peripheral Stent-Graft was placed under angiographic control, excluding the aneurysm from the circulation. No peri- or postprocedural complications were observed. At 6 months follow-up, the endograft is patent, excluding the aneurysm. Endovascular treatment may represent an alternative to surgery, especially in the distal infraparenchymal location. PMID- 17086456 TI - Endovascular treatment of ruptured iliac aneurysm previously treated by endovascular means. AB - A patient with a ruptured iliac aneurysm was admitted to the Emergency Department in hypovolemic shock. He had previously undergone surgical treatment for an infrarenal abdominal aortic aneurysm, which was managed with a terminal-terminal Dacron tube graft. Subsequently, he developed two iliac aneurysms, which were treated endovascularly with two wall-grafts in the right and one wall-graft in the left iliac arteries. He suffered chronic renal failure and arterial hypertension. Contrast-enhanced computed tomography showed rupture of the right iliac aneurysm and dislocation of the two wall-grafts. He was treated in an emergency situation with the implantation of an iliac endograft that bridged the two wall-grafts, which resulted in hemostasis and stabilization of his condition. Five days later, in an elective surgical situation, he was treated with the implantation of an aorto-uni-iliac endograft combined with a femoral-femoral bypass. He was discharged 5 days later in good condition. At the 4 year follow-up visit, the endoprosthesis remained in place with no evidence of an endoleak. In conclusion, overlapping of endografts should be avoided, if possible. Strict surveillance of the endovascularly treated patient remains mandatory. PMID- 17086457 TI - Comparison of reconstruction intervals in routine ECG-pulsed 64-row-MSCT coronary angiography in frequency controlled patients. AB - PURPOSE: In light of the increasing use and acceptance of multislice computed tomography (MSCT) coronary angiography it was the purpose of this study to compare reconstruction intervals used in a routine ECG-pulsed MSCT coronary artery angiography setting with frequency controlled patients. METHODS: Examinations were performed on a Siemens Somatom Sensation 64 scanner with a total of 110 ml of contrast agent and ECG pulsing (interval from 40% to 70%) after oral application of a beta-blocker if the heart rate was higher than 65 bpm. All human subjects were referred for the evaluation of suspected coronary artery disease. Coronary artery segments were evaluated by two experienced radiologists in a consensus reading. A ranking of diagnostic image quality (from 1 (no evaluation possible) to 5 (excellent image quality)) was statistically evaluated by Wilcoxon Signed Rank Test. RESULTS: In 45 patients (30 male, 15 female, age 63.8 +/- 12.1 years) we detected a significant advantage of the 60% reconstruction interval over 40%, 50%, and 70% (for each p < 0.05). In cases of sudden arrhythmia or movement during the scan, additional reconstruction intervals within the ECG-pulsed reconstruction intervals remained necessary for diagnosis. CONCLUSION: In a routine diagnostic setting with frequency controlled patients and ECG pulsing the 60% reconstruction interval can be considered superior for the initial diagnosis in 64-row multislice computed tomography coronary angiography. However, further information can be derived from various reconstruction intervals such as 40% and 70%. PMID- 17086458 TI - Bare stent implantation in iatrogenic dissecting pseudoaneurysm of the superior mesenteric artery. AB - Iatrogenic arterial dissection leading to the development of dissecting pseudoaneurysms of the superior mesenteric artery (SMA) is a rare complication of angiography. Surgical and endovascular treatment options exist for this important condition. We report a case of bare stent implantation in dissecting pseudoaneurysm of the SMA that developed after angiography in a patient with acute mesenteric ischemia. Although it is rarely published, iatrogenic arterial dissection causing pseudoaneurysm can occur after diagnostic and interventional angiography. Bare stent implantation in dissecting pseudoaneurysm of the SMA could be an advantageous endovascular treatment option in selected cases due its to potential preservation of important side branches of the SMA. PMID- 17086459 TI - Embolization for hemoptysis in chronic thromboembolic pulmonary hypertension: report of two cases and a review of the literature. AB - Hemoptysis is a known complication in patients with bronchial artery hypertrophy due to a variety of chronic pulmonary disorders. Bronchial artery hypertrophy is observed in most patients with chronic thromboembolic pulmonary hypertension (CTEPH), but surprisingly little is known about the incidence of hemoptysis in these patients. In this paper, we report on 2 patients with CTEPH and recurrent severe hemoptysis, who were treated by bronchial artery embolization. One patient recovered and 1 patient died as a consequence of the bleeding. A systematic review revealed 21 studies on the underlying pathology in 1,844 patients with moderate to severe hemoptysis. CTEPH was reported to be the cause of bleeding in 0.1% (n = 2), pulmonary arterial hypertension without chronic thromboembolic disease in 0.2% (n = 4), and acute pulmonary embolism in 0.7% (n = 12) of the patients. In contrast to this, 5 patients (6%) in our own series of 79 CTEPH patients suffered from moderate to severe hemoptysis requiring medical intervention. Severe hemoptysis appears to be an uncommon, but possibly underreported, life-threatening complication in CTEPH patients. As most CTEPH patients require life-long anticoagulants a therapeutic dilemma may ensue. Therefore, we propose that even mild hemoptysis in CTEPH patients warrants prompt evaluation, and treatment by embolization should be offered as first choice in CTEPH patients. PMID- 17086460 TI - Comparative genomic hybridization array analysis and real-time PCR reveals genomic copy number alteration for lung adenocarcinomas. AB - Genomic alterations in lung cancer tissues have been observed in various studies. To analyze the aberrations in the genome of lung cancer patients, we used array comparative genomic hybridization (array CGH) in 15 lung adenocarcinoma (AdC) tissues. Copy number gains and losses in chromosomal regions were detected and corresponding genes were confirmed by real-time polymerase chain reaction (PCR). As for the results, several frequently altered loci, including gain of 16p (46% of samples), were found, and the most common losses were found in 14q32.33 (26% of samples). High-level DNA amplifications (> 0.8 log(2) ratio) were detected at 1p, 5p, 7p, 9p, 11p, 11q, 12q, 14q, 16p, 17q, 19q, 20p, 21q, and 22q. A subset of genes, gained or lost, was checked for over- or underrepresentation by means of real-time PCR. The degree of fold change was highest in ECGF1 (22q13.33), HOXA9 (7p15.2), MAFG (17q25.3), TSC2 (16p13.3), and ICAM1 (19p13.2) genes and the 16p chromosome terminal region (16p13.3pter). Taken together, these results show that array CGH could be used as a powerful tool for identification of genomic alteration for lung cancer, and the above-mentioned genes may represent potential candidate genes in the study of lung cancer pathogenesis and diagnosis. PMID- 17086461 TI - Medical volunteering: giving something back. AB - A national health issue is how to provide medical care for the large number of people who are uninsured or do not qualify for medical coverage. Establishing free health clinics staffed by volunteer health professionals is one approach that is increasing, but this alone will not solve this societal problem. Many volunteer health care providers are needed, and more senior and retired physicians might be recruited. However, practicing general, not subspeciality, medicine in an unfamiliar surrounding with different patient demands may seem intimidating and anxiety producing. However, a conducive clinical environment and working with other volunteer health care staff may alleviate these feelings and make medical volunteering very enjoyable. The Mercy Health Clinic in Montgomery County, Maryland, has had this effect on us who volunteer there and care for its needy patients. PMID- 17086462 TI - Exercise-induced pulmonary hemorrhage after running a marathon. AB - We report on a healthy 26-year-old male who had an exercise-induced pulmonary hemorrhage (EIPH) within 24 hours of running a marathon. There were no symptoms, abnormalities on exam, or radiographic infiltrates. He routinely participated in bronchoscopy research and the EIPH was evident on gross inspection of bronchoalveolar lavage fluid, examination of the cytospin, and measurement of hemoglobin, iron, and ferritin concentrations. EIPH in humans may occur without any evidence on clinical presentation; its incidence may be far greater than currently suspected. PMID- 17086463 TI - N-95 face mask for prevention of bird flu virus: an appraisal of nanostructure and implication for infectious control. PMID- 17086464 TI - Preventive effect of Hochu-ekki-to on lipopolysaccharide-induced acute lung injury in BALB/c mice. AB - This study was designed to investigate the effect of Hochu-ekki-to (TJ-41), a Japanese herbal medicine, on the development of lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. ALI was induced in female BALB/c mice by the intranasal administration of 0.1 mg/kg LPS. The mice were divided into a group receiving normal feed and another group receiving feed mixed with TJ-41 at a dose of 1 g/kg/day for 8 weeks before LPS challenge. In the bronchoalveolar lavage fluid, the preadministration of TJ-41 caused significant reduction in the absolute number of total cells, neutrophils, and macrophages. The preadministration of TJ-41 significantly inhibited increases in the serum level of keratinocyte chemoattractant (KC), which is a murine chemotaxin for neutrophils that corresponds to human interleukin-8, with respect to its concentration at 24 h after LPS challenge. Furthermore, the histopathologic findings indicated that alveolitis with leukocyte infiltration in the alveolar space was less severe in the TJ-41-treated mice than in the control mice. These findings indicated that the preadministration of TJ-41 could show an inhibitory effect on ALI in this experimental murine system associated with the suppression of chemokine production. PMID- 17086465 TI - Comparison of 4 AM and 4 PM bronchial responsiveness to hypertonic saline in asthma. AB - Bronchial responsiveness to methacholine or histamine increases at night and may contribute to the mechanisms of nocturnal asthma. Hypertonic saline (HS) is a more clinically relevant stimulus for the diagnosis and assessment of the severity of asthma, but the circadian variation in bronchial responsiveness to hypertonic challenges has not been addressed. The aim of this study was to compare the responsiveness to hypertonic saline at 4:00 AM and at 4:00 PM. Eighteen diurnally active patients (11 women) with asthma, 31 +/- 9 years of age (mean +/- SD) and with a forced expiratory volume in 1 s (FEV(1)) of 79.11% +/- 12.85%, underwent two challenge tests (4:00 AM and 4:00 PM) in random sequence separated by an interval of 7 days. The challenge test consisted of inhalations of 4.5% saline with increasing doses by doubling the duration of nebulization (0.5, 1, 2, 4, and 8 min). The inhalation continued until a drop of 20% in FEV(1) was achieved or total time of 15.5 min. The provocative dose that caused the 20% drop in FEV(1) (PD(20)) was calculated. Differences were found between 4:00 PM and 4:00 AM values for inhalation times [3.80 +/- 3.57 min and 2.19 +/- 2.42 min (p = 0.001), respectively] and for PD(20) [4.94 +/- 6.77 ml and 2.93 +/- 4.74 ml (p = 0.002), respectively]. Eight patients with a home-assessed nocturnal peak expiratory flow (PEF) drop of more than 15% formed the nocturnal asthma group. The behavior of these patients was similar to that of the non-nocturnal asthma group. We conclude that the bronchial responsiveness to HS increases at night. PMID- 17086466 TI - Effect of histologic type on recurrence pattern in radiation therapy for medically inoperable patients with stage I non-small-cell lung cancer. AB - Japanese randomized trials showed that there was a significant impact on survival from stage I adenocarcinoma (AD) of the lung by adjuvant chemotherapy with uracil tegaful after complete resection but there was no effect for patients with squamous cell carcinoma (SQ). The purpose of this study was to examine the correlation of tumor histology and clinical outcome of radiation therapy (RT) for stage I non-small-cell lung cancer (NSCLC) and to consider the necessity of adjuvant chemotherapy after RT for these patients. The subjects were 83 patients, 54 with SQ and 29 with AD; they had received definitive RT with the total dose ranging from 60 to 80 Gy with conventional fractionation at a daily dose of 2 Gy. The differences between SQ and AD with respect to survival and recurrence pattern were investigated. The 5-year overall survival and cause-specific survival rates were 26.5% and 49.1%, respectively. No difference in survival was observed between SQ and AD patients, and the recurrence rates were almost identical (44% for SQ and 45% for AD). However, the 5-year primary control rate of SQ was significantly poorer than that of AD (SQ: 61.5%; AD: 87.6%; p = 0.03). Conversely, the 5-year metastasis-free survival rate of SQ was significantly better than that of AD (SQ: 88.2%; AD: 53.0%; p = 0.005). The different failure pattern, according to tumor histology, indicates that taking into consideration the difference in their clinical behaviors would also be important for planning RT and surgery for early lung cancer. PMID- 17086467 TI - Tracheal diverticulum: a review of the literature. AB - Tracheal diverticulum is a rarely encountered entity. It may be congenital or acquired, the difference residing mainly in the histologic features of the wall. Most cases are asymptomatic, but when symptoms are present they are usually nonspecific. Congenital diverticulum is not normally detected in infancy unless it is suggested by recurrent episodes of tracheobronchial infection or in associated with other malformations. Imaging techniques are useful for diagnosis because the point of communication with the trachea is difficult to detect with bronchoscopy. In the absence of symptoms, management should be conservative. We review the entity and report two new cases, one a casual finding in a 49-year-old man and the other in a 63-year-old woman who presented with occasional bloody sputum. PMID- 17086468 TI - Infection of an abdominal aortic stent graft with suprarenal attachment. AB - Stent grafting for treatment of abdominal aortic aneurysm can lead to infectious complications. The purpose of this report is to describe a case involving a patient with an aortobiliac stent graft, a horseshoe kidney, and coronary artery disease who presented suprarenal aortic infection requiring removal of the stent graft and in situ reconstruction using an arterial homograft. The discussion deals with management of infected vascular material and reconstruction techniques. PMID- 17086469 TI - Mid-term results of endovascular treatment for descending thoracic aorta diseases in high-surgical risk patients. AB - We report the initial experience of two cardiovascular surgery centers in the treatment of descending thoracic aorta lesions with covered stent grafts in high surgical risk patients. From April 1999 to November 2004, 54 patients, mean age 64 years (range 16-83), were treated by stent graft for a lesion of the descending aorta (degenerative aneurysms n = 22, aortic dissections n = 12, chronic post traumatic aneurysms n = 5, anastomotic false aneurysms n = 2, penetrating ulcers n = 4, intramural hematomas n = 5, traumatic rupture n = 4), with 42.6% treated on an emergency basis. Three devices were used: Talent (n = 49), Excluder (n = 4), and Zenith (n = 1). In three patients, combined surgery of the proximal aorta was performed. Prior bypass of the left supra-aortic arteries was performed in four patients. The follow-up was clinical and radiological (plain chest film and computed tomographic scan) at 1, 3, 6, 12, 18, and 24 months and yearly thereafter. The stent graft was successfully deployed in all cases. Two early deaths related to the stent graft (one migration and aortic rupture and one stroke) and one related to adult respiratory distress syndrome occurred. Morbidity was 16.6% (iliac access damage n = 4, groin reintervention n = 3, transient ischemic attack n = 1, tamponade n = 1). The follow-up was 100% complete (mean 22.8 months, range 3-51). Fifteen primary endoleaks (type I n = 6, type II n = 8, type III n = 1) and one secondary endoleak were reported. They were treated by additional stent graft (n = 7) and elective surgical conversion (n = 1). Six endoleaks resolved spontaneously at 6 months, and two are being monitored. Twelve endoleaks (75%) occurred in patients treated for degenerative aneurysms. Freedom from secondary reintervention was 81.3% at 3 years. Two transient paraparesias were observed at 3 and 18 months. Of the 13 deaths observed during the follow-up, only one was related to the stent graft. Actuarial survival at 12 and 24 months was 90.0% and 75.4%, respectively. Mortality results are encouraging in this specific cohort of high-surgical risk patients. A new kind of morbidity is observed, related to endoleaks, whose necessary management could hinder the durability of the technique. PMID- 17086470 TI - Persistence, reproducibility, and cost-effectiveness of an intervention to improve the quality of osteoporosis care after a fracture of the wrist: results of a controlled trial. AB - INTRODUCTION: Older patients with fragility fractures are not commonly tested or treated for osteoporosis. Compared to usual care, a previously reported intervention led to 30% absolute increases in osteoporosis treatment within 6 months of wrist fracture. Our objective was to examine longer-term outcomes, reproducibility, and cost-effectiveness of this intervention. METHODS: We conducted an extended analysis of a non-randomized controlled trial with blinded ascertainment of outcomes that compared a multifaceted intervention to usual care controls. Patients >50 years with a wrist fracture treated in two Emergency Departments in the province of Alberta, Canada were included; those already treated for osteoporosis were excluded. Overall, 102 patients participated in this study (55 intervention and 47 controls; median age: 66 years; 78% were women). The interventions consisted of faxed physician reminders that contained osteoporosis treatment guidelines endorsed by opinion leaders and patient counseling. Controls received usual care; at 6-months post-fracture, when the original trial was completed, all controls were crossed-over to intervention. The main outcomes were rates of osteoporosis testing and treatment within 6 months (original study) and 1 year (delayed intervention) of fracture, and 1-year persistence with treatments started. From the perspective of the healthcare payer, the cost-effectiveness (using a Markov decision-analytic model) of the intervention was compared with usual care over a lifetime horizon. RESULTS: Overall, 40% of the intervention patients (vs. 10% of the controls) started treatment within 6 months post-fracture, and 82% (95%CI: 67-96%) had persisted with it at 1-year post-fracture. Delaying the intervention to controls for 6 months still led to equivalent rates of bone mineral density (BMD) testing (64 vs. 60% in the original study; p = 0.72) and osteoporosis treatment (43 vs. 40%; p = 0.77) as previously reported. Compared with usual care, the intervention strategy was dominant - per patient, it led to a $13 Canadian (U.S. $9) cost savings and a gain of 0.012 quality-adjusted life years. Base-case results were most sensitive to assumptions about treatment cost; for example, a 50% increase in the price of osteoporosis medication led to an incremental cost-effectiveness ratio of $24,250 Canadian (U.S. $17,218) per quality-adjusted life year gained. CONCLUSIONS: A pragmatic intervention directed at patients and physicians led to substantial improvements in osteoporosis treatment, even when delivered 6-months post-fracture. From the healthcare payer's perspective, the intervention appears to have led to both cost-savings and gains in life expectancy. PMID- 17086473 TI - Morbidity in 201 patients with small sized meningioma treated by microsurgery. AB - BACKGROUND: The management of patients with small, often asymptomatic meningiomas is controversial and includes observation, microsurgery (MS) and stereotactic radiosurgery (SRS). The purpose of this retrospective study was to analyze the morbidity and the extent of removal after MS for small (< or =3 cm) intracranial meningiomas and compare these results to those of SRS reported in the literature. METHODS: All patients with an intracranial meningioma with a maximum diameter up to 3 cm operated on in our institution over a 10 year period (1992-2002) were included in the study and retrospectively analyzed. Patients were grouped into asymptomatic and symptomatic and according to tumor location as: group I (cranial vault, parasagittal, lateral sphenoid), group II (falx, frontobasal, medial sphenoid, parasellar and tentorial), group III (cavernous sinus, petroclival, petrosal, CPA and foramen magnum). FINDINGS: There were a total of 201 patients, of whom 102 were asymptomatic and 99 were symptomatic. The overall risk of permanent neurological morbidity was 4.9% in asymptomatic and 23.2% in symptomatic patients. The combined risk in asymptomatic and symptomatic patients was 5.4% in group I, 11.5% in group II, and 39.9% in group III lesions. Radical removal was achieved in all patients in group I, in 93.7% of group II, and 80% of group III lesions. There was no disease related mortality. CONCLUSIONS: MS provides excellent efficacy and morbidity results in groups I and II meningiomas, especially in asymptomatic patients and might therefore be considered the first choice of treatment for these patients. The results of MS in group III were worse than those of SRS reported in the literature. PMID- 17086474 TI - Hydrocephalus in Paget's disease. AB - Neurological manifestations in Paget's disease are rare. Hydrocephalus is infrequent, its etiology in Paget's disease being multi-factorial. Surgical and anesthetic procedures may be challenging, requiring special precautions. We describe one such case and discuss these peculiarities. An elderly lady, diagnosed to have Paget's disease presented with features of recent onset dementia, gait disturbances with urinary incontinence. Hydrocephalus secondary to posterior fossa crowding was present. A ventriculo-peritoneal shunt was inserted. Endotracheal intubation proved to be difficult. Modification of the surgical technique was required to ensure a safe procedure. She improved immediately following CSF diversion. Timely CSF diversion yields good results. Certain precautions during anesthesia and surgery can ensure a safe and uneventful peri operative period. PMID- 17086475 TI - Tentorial meningioma on follow-up presenting with sudden deterioration due to intra- and peritumoral hemorrhage. AB - Almost all meningiomas presenting with intracranial hemorrhage in the literature were admitted with symptoms relating to the hemorrhage and there were no prehemorrhage scans which demonstrated the actual size and apperance of the meningioma. This is a very rare report of a case with a tentorial meningioma documented with pre- and posthemorrhage scans. PMID- 17086476 TI - Objective criteria for successful transsphenoidal removal of suprasellar nonfunctioning pituitary adenomas. A prospective study. AB - BACKGROUND: Despite ample experience with transsphenoidal surgery, objective data on which suprasellar tumour expansion and growth pattern allows for radical adenoma resection are still sparse. Hence, we have performed a prospective study to establish the predictive value of tumour dimension and shape for the intra operative descent of the diaphragma, the completeness of tumour resection and the outcome of patients harbouring pituitary adenomas with suprasellar extension. METHOD: Included in the study were 105 patients with nonfunctioning pituitary adenomas and suprasellar extension who underwent primary transsphenoidal surgery between January 1998 and December 2005. The precise suprasellar extension, the degree of dumbbell-shape, the configuration of the adenomas and the depth of the pituitary fossa were evaluated. Completeness of resection was assessed by MRI at 3 months postoperatively. FINDINGS: The mean cranio-caudal diameter of the tumours was 28.0 mm (range 9.2-57.8 mm). On average, the suprasellar extension measured 11.9 mm (range 2.1-25.8 mm). Total removal of the suprasellar tumour was accomplished in 83% (87 of 105) of the patients. A second operation for residual adenoma was only indicated in 2 cases. The vertical intracranial extension was the strongest independent predictor of subtotal resection (p < 0.001). Irregular and multilobular configuration was a second highly-significant and independent predictor for incomplete resection (p < 0.003). In contrast, dumbbell-shape and shallow pituitary fossa were not independent predictive factors for incomplete tumour resection. The complication rate was very low. None of our patients suffered postoperative rhinorrhea, meningitis or visual deterioration. CONCLUSIONS: One-stage transsphenoidal surgery allows total or near-total resection of most suprasellar pituitary adenomas with low surgical morbidity. Quantitative assessment of tumour dimension and configuration contributes to establishing guidelines for the selection of the appropriate approach and prediction of surgical outcome. PMID- 17086477 TI - Profiling signalling pathways of the receptor activator of NF-kappaB ligand induced osteoclast formation in mouse monocyte cells, RAW264.7. AB - Cell-based signal chemical genomics can profile the signalling pathway for certain cellular events by using a target-known chemical library. To ascertain its usefulness, the receptor activator of NF-kappaB ligand (RANKL)-induced osteoclastogenesis in mouse monocyte/macrophage cells RAW264.7 was used as an in vitro experimental model. Of 180 target-known inhibitors/activators formatted in a 384-well plate, 8 chemicals were shown to inhibit the osteoclast formation, but 4 chemicals enhanced this process. A variety of references support, or possibly lead one to expect the effects of these 12 chemicals on the cellular process of osteoclastogenesis in RAW264.7 cells, but several signalling pathways were newly found in this study; for example, CA-074 Me inhibiting cathepsin B and nitrendipine blocking the calcium channel could have the potential to inhibit the osteoclast formation as well as bone resorption. This is a simple but very fast and powerful method of profiling the signalling pathway of certain cellular events. Signal chemical genomics could provide invaluable information for the exploration of new target signalling processes and further target-based drug discovery strategies. PMID- 17086478 TI - Effects of ligands on the stability of tissue transglutaminase: studies in vitro suggest possible modulation by ligands of protein turn-over in vivo. AB - Tissue transglutaminase catalyzes irreversible post-translational modification of specific protein substrates by either crosslinkage or incorporation of primary amines into glutamine residues, through glutamyl-amide isopeptide bonds. Modulation in vivo of these reactions (collectively called "transamidation") is brought about by both ligand dependent effects (chiefly, activation by calcium and inhibition by GTP) as well as by variation in enzyme tissue levels by transcriptional effects. Accumulating observations that the enzyme stability in vitro is greatly affected by interaction with ligands led us to postulate that also the turn-over in vivo might be modulated by ligands opening new scenarios on the regulation of the tissue transamidating activity. This proposal is consistent with data obtained in in vitro cell culture systems and has important implications for the expression of activity in vivo. PMID- 17086479 TI - Effect of beta-alanine administration on carbon tetrachloride-induced acute hepatotoxicity. AB - Mice were supplemented with beta-alanine (3%) in drinking water for one week. beta-Alanine intake reduced hepatic taurine levels, but elevated cysteine levels significantly. Hepatotoxicity of CCl4 in mice fed with beta-alanine was decreased as determined by changes in serum enzyme activities. Hepatic glutathione and taurine concentrations after CCl4 challenge were increased markedly by beta alanine intake. The enhanced availability of cysteine for synthesis of glutathione and/or taurine appears to account for the hepatoprotective effects of beta-alanine against CCl4-induced liver injury. PMID- 17086480 TI - Disturbance of erythrocyte lipid bilayer by amino acid-based surfactants. AB - In an attempt to increase our knowledge regarding the mechanisms of surfactant membrane interaction, we studied the action of several anionic and cationic amino acid-based surfactants on membrane fluidity using fluorescence anisotropy. Anisotropy measurements demonstrated that almost all of the surfactants studied disturbed the external region of the erythrocyte membrane without affecting the core of the bilayer. How the physico-chemical properties and structure of these compounds affect dynamics of the lipid bilayer is discussed in detail. PMID- 17086481 TI - Free radical scavenging potential of L-proline: evidence from in vitro assays. AB - An assessment of the potential of proline to scavenge free radicals was made in a couple of in vitro assay systems, namely graft co-polymerization and autooxidation of pyrogallol. Both these assays are essentially dependent upon free radical mechanisms. Graft co-polymerization involved a ceric (Ce(4+)) ion- or gamma-radiation-induced grafting of methyl acrylate (MA) onto a cellulose backbone. The degree of grafting, measured gravimetrically, was taken as a measure of free radical generation. The gamma-radiation-dependent grafting was far greater than that due to Ce(4+) ions. Inclusion of proline in the assay, irrespective of the initiator used, led to suppression of grafting in a concentration-dependent manner indicating the ability of proline to scavenge free radicals. The gamma-radiation-dependent grafting was also suppressed by hydroquinone and glutathione but not by ascorbate, glycine and spermine. In contrast to graft co-polymerization, proline did not inhibit the autooxidation of pyrogallol, a reaction involving superoxide radical generation. A subset of data constitutes an evidence for the ability of proline to scavenge free radicals in vitro. It is implied by extension that free proline, known to accumulate in plant tissues during abiotic stresses, would contribute to scavenging of surplus free radicals produced under a variety of abiotic stresses. PMID- 17086482 TI - Gunshot wounds to the chest with arterial bullet embolization. AB - This report describes two cases involving small-caliber gunshot wounds to the chest with embolization of the bullet (.22 long rifle). In the first patient, hemodynamic status permitted additional imaging before surgery. Findings demonstrated embolization of the bullet into the right external carotid artery after penetration into the left ventricle. In the second patient, emergency thoracotomy was required to perform hemostasis following perforation of the pulmonary artery with massive hemothorax. Bullet embolization into the left hypogastric artery through an acquired aortopulmonary fistula suspected during thoracic exploration was diagnosed after the procedure. Postoperative recovery was uneventful in both cases. The bullet in the hypogastric artery was not extracted, with no complications being observed after a follow-up period of 3 years. PMID- 17086483 TI - Cerebral ischemia during carotid artery cross-clamping: predictive value of phase contrast magnetic resonance imaging. AB - The goal of this prospective study was to determine the utility of preoperative cerebral magnetic resonance imaging (MRI) in predicting cerebral ischemia during carotid artery cross-clamping for endarterectomy. Between January 2000 and December 2003, a total of 121 patients (95 men, 26 women) underwent three dimensional phase-contrast MRI to assess collateral function prior to carotid endarterectomy. During regional anesthesia, patients were monitored to detect ischemic events and their timing in relation to cross-clamping and to determine mean intraoperative arterial pressure. These findings were then correlated with the collateral variations observed in the circle of Willis on preoperative MRI. Patients were classified into three groups according to neurological tolerance: normal tolerance (n = 106), immediate severe deficit (n = 9), and late deficit associated with arterial hypotension (n = 6). In the second group, a significant correlation was found between the absence of collateral circulation and neurological deficit (p < .0001). These results indicated that three-dimensional phase-contrast MRI is useful for predicting cerebral ischemia during carotid cross-clamping and selecting indications for shunting. Absence of visible collaterals of the circle of Willis on MRI is significantly predictive of early ischemia and an indication for systematic shunt placement. PMID- 17086484 TI - Nasal carriage of methicillin-resistant staphylococcus aureus in vascular surgery. AB - The purpose of this study was to determine the prevalence of nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA) and to define risk factors allowing identification of high-risk patients for MRSA nasal carriage at admission to the vascular surgery unit. From March 23, 2004 to July 13, 2004, screening for nasal carriage of MRSA was conducted at admission to the vascular surgery unit and 1 week thereafter. To analyze risk factors for MRSA nasal carriage at admission to the vascular surgery unit, a case-control study was carried out in patients presenting colonization at the time of admission. A total of 308 patients underwent nasal screening for MRSA. Thirteen were colonized with MRSA (nine at admission and four acquired), i.e., 4.2% of patients. Methicillin susceptible Staphylococcus aureus (MSSA) was found in 11.4% of patients who underwent screening. Six patients with MRSA infection were identified during the study period. The two patients who acquired infection were colonized at the time of admission. Arrival from another health-care facility and from another department was a significant risk factor for carriage of MRSA. The prevalence of nasal carriage in vascular surgery was 4.2%. Nasal screening is highly cost effective since 60% of MRSA carriers were undetected using diagnostic specimens alone. French recommendations issued for cardiac and orthopedic surgery by the consensus conference on preoperative management of infectious risk on March 5, 2004, should be extended to vascular surgery. PMID- 17086485 TI - Utilizing program evaluation as a strategy to promote community change: evaluation of a comprehensive, community-based, family violence initiative. AB - This paper describes the authors' work in a community that received Federal funding for an integrated system of care to reduce the impact and incidence of exposure to violence for children less than six years of age. The paper includes a review of the conceptual framework that guided the work of the authors and provides a brief overview of the issue of family violence, the impact of this violence on young children, and the Federal response to this issue. In addition, a description of the Initiative and the community in which it was based is provided along with some aspects of the evaluation plan. Finally, the authors discuss how their work with this Initiative depicts an approach to facilitating change within communities. PMID- 17086486 TI - Endothelial calcium signaling in rabbit arteries and its local alterations in early-stage atherosclerosis. AB - This study is to examine whether endothelial calcium signaling is different between atherosclerosis-prone thoracic aortas (TA) and atherosclerosis-resistant carotid arteries (CA) in normal rabbits and how it changes in early-stage atherosclerosis. Local endothelial calcium signaling was examined in arterial segments obtained from rabbits fed with normal or high-cholesterol diet for 1-4 weeks. Contrasting to normal CA, normal TA showed lower endothelial calcium signaling with more concentrated NF-kappaB in the endothelial nuclei. In the same hypercholesterolemic animal, fatty streak formation was much more prominent in TA than in CA. TA endothelial calcium signaling became augmented in the second week of hypercholesterolemia, being most pronounced in smooth regions adjacent to miniature fatty streaks. It was sporadically elevated even in regions away from any detectable TA fatty streak. When the entire TA was covered with fatty streaks in the fourth week of hypercholesteremia, endothelial calcium signaling returned to the original level. In comparison, CA endothelial calcium signaling was reduced around scattered fatty streaks. Reduced calcium signaling happened where CA fatty streaks were 150 microm long (covering 15-30 cells); and it extended to areas adjacent to larger fatty streaks. Moreover, NF-kappaB remained in the cytosol of endothelial cells covering CA fatty streaks. Our results indicate that inter-vascular differences in endothelial calcium signaling may provide partial explanation in their differential susceptibility in atherosclerosis. PMID- 17086487 TI - Gender- and region-specific expression of insulin receptor protein in mouse brain: effect of mild inhibition of oxidative phosphorylation. AB - Insulin receptors (IR) and inhibition of oxidative metabolism have been suggested to partake in the pathophysiological cascade of neurodegenerative disorders. The goal of this study was to investigate gender- and region-specificity of insulin receptor protein expression in mouse brain subsequent to a mild hypoxic episode. Tissue was prepared from untreated male and female mice and animals pretreated in vivo with 20 mg/kg body weight i.p. 3-nitroproprionic acid (3-np; an inhibitor of succinic dehydrogenase) 1 hr prior to tissue preparation. IR expression in control animals was alike in males and females during proestrus and estrus but reduced during diestrus. On pretreatment, IR protein expression decrease in hippocampus in males but remained alike in other regions and females. In summary, IR protein expression is regionally different in males and females, gender dependent, and modulated during the stages of the estrus cycle in females. Contrary to expectations it is not modified on mild inhibition of oxidative phosphorylation in any region in females and altered in hippocampus solely in males. The latter effect, however, warrants further scrutiny concerning participation in pathophysiological cascades affecting the hippocampus such as in Alzheimer's disease. PMID- 17086488 TI - The role of FDG-PET in the selection of patients with colorectal liver metastases. AB - BACKGROUND: Selection of patients for hepatic resection of colorectal liver metastases is still limited. After conventional work up by computed tomography (CT) scan, 60% of patients will develop recurrent disease in the early years after resection. The aim of the present study was to evaluate whether an additional fluorine-18-deoxyglucose positron emission tomography (FDG-PET) improves patient selection and therefore adds value to select patients for curative liver resection. METHODS: Data from 203 patients selected for surgical treatment of colorectal liver metastases between 1995 and 2003 were collected in a prospective database. Group A consisted of 100 consecutive patients selected for hepatic surgery by conventional diagnostic imaging (CT chest and abdomen) only. Group B consisted of 103 consecutive patients selected for hepatic surgery by conventional diagnostic methods plus an additional FDG-PET. RESULTS: The number of patients with futile surgery, in which further treatment was considered inappropriate at laparotomy, was 28.0% in group A and 19.4% in group B. The reason for unresectable disease differed between groups. In group A, 10/100 (10.0%) patients showed extrahepatic abdominal disease versus 2/103 patients (1.9%) in group B (P = .017). In all other cases, resection was not performed because liver disease proved too extensive at laparotomy. For patients ultimately undergoing surgical treatment of the metastases, survival was comparable between groups. Overall survival at 3 years was 57.1% in group A versus 60.1% in group B. Disease-free survival at 3 years was 23.0% in group A and 31.4% in group B. CONCLUSIONS: In patients with colorectal liver metastases, FDG-PET may reduce the number of negative laparotomies. However, the effect size on the selection of these patients seems not sufficient enough to affect the overall and disease-free survival after treatment. PMID- 17086489 TI - The modeling of global epidemics: stochastic dynamics and predictability. AB - The global spread of emergent diseases is inevitably entangled with the structure of the population flows among different geographical regions. The airline transportation network in particular shrinks the geographical space by reducing travel time between the world's most populated areas and defines the main channels along which emergent diseases will spread. In this paper, we investigate the role of the large-scale properties of the airline transportation network in determining the global propagation pattern of emerging diseases. We put forward a stochastic computational framework for the modeling of the global spreading of infectious diseases that takes advantage of the complete International Air Transport Association 2002 database complemented with census population data. The model is analyzed by using for the first time an information theory approach that allows the quantitative characterization of the heterogeneity level and the predictability of the spreading pattern in presence of stochastic fluctuations. In particular we are able to assess the reliability of numerical forecast with respect to the intrinsic stochastic nature of the disease transmission and travel flows. The epidemic pattern predictability is quantitatively determined and traced back to the occurrence of epidemic pathways defining a backbone of dominant connections for the disease spreading. The presented results provide a general computational framework for the analysis of containment policies and risk forecast of global epidemic outbreaks. PMID- 17086490 TI - Fixation of strategies for an evolutionary game in finite populations. AB - A stochastic evolutionary dynamics of two strategies given by 2x 2 matrix games is studied in finite populations. We focus on stochastic properties of fixation: how a strategy represented by a single individual wins over the entire population. The process is discussed in the framework of a random walk with site dependent hopping rates. The time of fixation is found to be identical for both strategies in any particular game. The asymptotic behavior of the fixation time and fixation probabilities in the large population size limit is also discussed. We show that fixation is fast when there is at least one pure evolutionary stable strategy (ESS) in the infinite population size limit, while fixation is slow when the ESS is the coexistence of the two strategies. PMID- 17086491 TI - Interactions between pattern formation and domain growth. AB - In this paper we develop a theoretical framework for investigating pattern formation in biological systems for which the tissue on which the spatial pattern resides is growing at a rate which is itself regulated by the diffusible chemicals that establish the spatial pattern. We present numerical simulations for two cases of interest, namely exponential domain growth and chemically controlled growth. Our analysis reveals that for domains undergoing rapid exponential growth dilution effects associated with domain growth influence both the spatial patterns that emerge and the concentration of chemicals present in the domain. In the latter case, there is complex interplay between the effects of the chemicals on the domain size and the influence of the domain size on the formation of patterns. The nature of these interactions is revealed by a weakly nonlinear analysis of the full system. This yields a pair of nonlinear equations for the amplitude of the spatial pattern and the domain size. The domain is found to grow (or shrink) at a rate that depends quadratically on the pattern amplitude, the particular functional forms used to model the local tissue growth rate and the kinetics of the two diffusible species dictating the resulting behaviour. PMID- 17086492 TI - Effects of heterogeneity on spread and persistence in rivers. AB - The question how aquatic populations persist in rivers when individuals are constantly lost due to downstream drift has been termed the "drift paradox." Recent modeling approaches have revealed diffusion-mediated persistence as a solution. We study logistically growing populations with and without a benthic stage and consider spatially varying growth rates. We use idealized hydrodynamic equations to link river cross-sectional area to flow speed and assume heterogeneity in the form of alternating patches, i.e., piecewise constant conditions. We derive implicit formulae for the persistence boundary and for the dispersion relation of the wave speed. We explicitly discuss the influence of flow speed, cross-sectional area and benthic stage on both persistence and upstream invasion speed. PMID- 17086493 TI - On the population dynamics of the malaria vector. AB - A deterministic differential equation model for the population dynamics of the human malaria vector is derived and studied. Conditions for the existence and stability of a non-zero steady state vector population density are derived. These reveal that a threshold parameter, the vectorial basic reproduction number, exist and the vector can established itself in the community if and only if this parameter exceeds unity. When a non-zero steady state population density exists, it can be stable but it can also be driven to instability via a Hopf Bifurcation to periodic solutions, as a parameter is varied in parameter space. By considering a special case, an asymptotic perturbation analysis is used to derive the amplitude of the oscillating solutions for the full non-linear system. The present modelling exercise and results show that it is possible to study the population dynamics of disease vectors, and hence oscillatory behaviour as it is often observed in most indirectly transmitted infectious diseases of humans, without recourse to external seasonal forcing. PMID- 17086494 TI - The role of counter-current exchange in preventing hypoxia in skeletal muscle. AB - Mathematical models that describe oxygen transport from a single capillary into a region of surrounding tissue often predict that the tissue is hypoxic, whereas in reality diffusion from more richly perfused nearby capillaries prevents hypoxia from forming in the tissue. In this manuscript, a mathematical model of oxygen transport is presented that is applicable to vascular beds consisting of a large number of non-uniformly perfused parallel capillaries arranged in a manner characteristic of skeletal muscle. The model is used to examine conditions under which counter-current flow and myoglobin-facilitated diffusion provides sufficient oxygen to poorly perfused regions to prevent the occurrence of hypoxia. The method developed here leads to a coupled system of nonlinear ordinary differential equations for the oxygen concentration in the capillaries, and is easy to apply even for vascular beds containing a large number of capillaries. PMID- 17086495 TI - Calcium oscillations and waves generated by multiple release mechanisms in pancreatic acinar cells. AB - We explore the dynamic behavior of a model of calcium oscillations and wave propagation in the basal region of pancreatic acinar cells [Sneyd, J., et al., Biophys. J. 85: 1392-1405, 2003]. Since it is known that two principal calcium release pathways are involved, inositol trisphosphate receptors (IPR) and ryanodine receptors (RyR), we study how the model behavior depends on the density of each receptor type. Calcium oscillations can be mediated either by IPR or RyR. Continuous increases in either RyR or IPR density can lead to the appearance and disappearance of oscillations multiple times, and the two receptor types interact via their common effect on cytoplasmic calcium concentration and the subsequent effect on the total amount of calcium inside the cell. Increases in agonist concentration can stimulate oscillations via the RyR by increasing calcium influx. Using a two time-scale approach, we explain these complex behaviors by treating the total amount of cellular calcium as a slow parameter. Oscillations are controlled by the shape of the slow manifold and where it intersects the nullcline of the slow variable. When calcium diffusion is included, the existence of traveling waves in the model equation is strongly dependent on the interplay between the total amount of calcium in the cell and membrane transport, a feature that can be experimentally tested. Our results help us understand the behavior of a model that includes both receptors in comparison to the properties of each receptor type in isolation. PMID- 17086496 TI - Modeling intercellular interactions in early Mycobacterium infection. AB - Infection with Mycobacterium tuberculosis (Mtb) is characterized by localized, roughly spherical lesions within which the pathogen interacts with host cells. Containment of the infection or progression of disease depends on the behavior of individual cells, which, in turn, depends on the local molecular environment and on contact with neighboring cells. Modeling can help us understand the nonlinear interactions that drive the overall dynamics in this system. Early events in infection are particularly important, as are spatial effects and inherently stochastic processes. We describe a model of early Mycobacterium infection using the CyCells simulator, which was designed to capture these effects. We relate CyCells simulations of the model to several experimental observations of individual components of the response to Mtb. PMID- 17086497 TI - Modelling hematopoiesis mediated by growth factors with applications to periodic hematological diseases. AB - Hematopoiesis is a complex biological process that leads to the production and regulation of blood cells. It is based upon differentiation of stem cells under the action of growth factors. A mathematical approach of this process is proposed to understand some blood diseases characterized by very long period oscillations in circulating blood cells. A system of three differential equations with delay, corresponding to the cell cycle duration, is proposed and analyzed. The existence of a Hopf bifurcation at a positive steady-state is obtained through the study of an exponential polynomial characteristic equation with delay-dependent coefficients. Numerical simulations show that long-period oscillations can be obtained in this model, corresponding to a destabilization of the feedback regulation between blood cells and growth factors, for reasonable cell cycle durations. These oscillations can be related to observations on some periodic hematological diseases (such as chronic myelogenous leukemia, for example). PMID- 17086498 TI - Combined effect of 25-hydroxycholesterol and IL-1beta on IL-8 production in human colon carcinoma cell line (Caco-2). AB - Interleukin-1 beta (IL-1beta) is an important mediator in intestinal inflammation. IL-1beta promotes IL-8 production, which can be modulated by a number of factors, including oxidative stress. Interestingly, oxysterols, which are thought to contribute to inflammation in atherosclerotic plaques, are also produced by intestinal epithelial cells. Thus, we investigated the effect of oxysterols, including 25-hydroxycholesterol and 7beta-hydroxycholesterol, on IL 1beta-induced IL-8 production in Caco-2 cells (a human colon carcinoma cell line). Pre-treatment of Caco-2 cells with 25-hydroxycholesterol significantly enhanced IL-1beta-induced IL-8 expression at both mRNA and protein levels. However, 7beta-hydroxycholesterol showed very little effect on IL-8 production. Furthermore, pre-treatment with 25-hydroxycholesterol, followed by IL-1beta stimulation, enhanced IL-8 promoter activity beyond that observed with IL-1beta alone. These results suggest that 25-hydroxycholesterol enhances IL-1beta-induced IL-8 production, possibly by enhancing promoter activity. PMID- 17086499 TI - Reducing sickness absence from work due to low back pain: how well do intervention strategies match modifiable risk factors? AB - OBJECTIVE: To assess, from the review literature, the extent to which effective strategies for reducing work absence after acute low back pain (LBP) match empirical risk factors. METHODS: From 17 recent review articles (2000-2005), disability risk factors and interventions were cross-tabulated to assess levels of relative concordance. RESULTS: Potentially modifiable risk factors included 23 variables describing 3 workplace and 3 personal domains. Effective interventions included 25 strategies that were personal (physical or behavioral), engineering, or administrative in nature. There was a strong risk factor concordance for workplace technical and organizational interventions, graded activity exposure, and cognitive restructuring of pain beliefs. There was less risk factor concordance for exercise, back education, and RTW coordination. Few interventions focused on relieving emotional distress or improving job dissatisfaction, two well-supported risk factors. DISCUSSION: Gaps between the epidemiological and intervention research of back disability prevention could be reduced by testing mediators of intervention effects or by stratifying outcomes according to pre intervention risk factors. PMID- 17086500 TI - Effects of a proximity-sensing feedback chair on head, shoulder, and trunk postures when working at a visual display terminal. AB - INTRODUCTION: This study was designed to identify the effects of feedback from a proximity-sensing chair on head, shoulder, and trunk postures when working at a visual display terminal (VDT). METHODS: Twenty healthy adults were asked to perform VDT work, and their forward head, forward shoulder, and trunk flexion angles were analyzed using a 3-D motion analysis system. The statistical significance of differences between without and with an auditory feedback device was tested by paired t-tests, with the significance cutoff set at alpha=0.05. RESULTS: The forward head, forward shoulder, and trunk flexion angles significantly decreased during VDT work when using the proximity sensor with auditory feedback. CONCLUSION: We suggest that a feedback device promotes the adoption of beneficial postures, which may be effective in preventing VDT-work related neck and upper-limb disorders. PMID- 17086501 TI - A comparison of two lifting assessment approaches in patients with chronic low back pain. AB - The Progressive Isoinertial Lifting Evaluation (PILE) and the lifting test of the WorkWell Systems Functional Capacity Evaluation (WWS) are well known as lifting performance tests. The objective of this study was to study whether the PILE and the WWS can be used interchangeably in patients with Chronic Low Back Pain (CLBP) and to explore whether psychosocial variables can explain possible differences. METHODS: 53 Patients (32 men and 21 women) with CLBP were tested twice in a counter balanced design. Pearson Correlation Coefficient of r > 0.75 and non significant differences on two-tailed t tests were considered as good comparability. RESULTS: Pearson Correlation Coefficient was 0.75 (p < 0.01). Lifting performance on the WWS was a mean of 6.0 kg higher compared to the PILE (p < 0.01). The difference between the PILE and the WWS was unrelated to psychological variables. CONCLUSION: It can be concluded that the PILE and the WWS cannot be used interchangeably. Psychosocial variables cannot explain the differences between both tests. PMID- 17086502 TI - The effect of alternate style keyboards on severity of symptoms and functional status of individuals with work related upper extremity disorders. AB - There is evidence that performing job tasks involving repetition, vibration, sustained posture or forceful movement may contribute to symptoms of work related upper extremity disorders. Typing is one such activity; symptoms that develop as a result of this activity can affect performance of work, self-care and leisure occupations. Studies investigating the impact of ergonomic keyboards on symptom reduction are limited, and little research exists regarding the reduction of key activation force as an intervention. METHODS: This randomized, prospective study used a sample of 68 symptomatic workers employed by a single company. One group received a commercially available ergonomic keyboard, a second group used a modified version of the same keyboard designed to reduce activation force, vibration and key travel. We measured symptoms and clinical signs, functional status, and device satisfaction in both groups over a six-month study period. CONCLUSIONS: Between-groups analyses indicated that the groups performed similarly on the outcomes of interest. Repeated-measure analysis identified a reduction of symptoms, an improvement in functional status, preference for and increased satisfaction with the intervention keyboards, and maintenance of typing speed and accuracy for both groups. PMID- 17086503 TI - Economic evaluation of a multi-stage return to work program for workers on sick leave due to low back pain. AB - OBJECTIVE: To evaluate the cost-effectiveness and cost-utility of a return to work (RTW) program for workers on sick-leave due to low back pain (LBP), comparing a workplace intervention implemented between 2 to 8 weeks of sick-leave with usual care, and a clinical intervention after 8 weeks of sick-leave with usual care. DESIGN: Economic evaluation alongside a randomised controlled trial (RCT). STUDY POPULATION: Workers sick-listed for a period of 2 to 6 weeks due to LBP. INTERVENTIONS: 1. workplace assessment, work modifications and case management). 2. physiotherapy based on operant behavioural principles. 3. usual care: provided by an occupational physician. OUTCOMES: The primary outcome was return to work (RTW). Other outcomes were pain intensity, functional status, quality of life and general health. The economic evaluation was conducted from a societal perspective. Outcomes were assessed at baseline (after 2-6 weeks on sick leave), and 12 weeks, 26 weeks, and 52 weeks after the first day of sick-leave. RESULTS: The workplace intervention group returned to work 30.0 days (95% CI=[3.1, 51.3]) earlier on average than the usual care group at slightly higher direct costs (ratio of 1 day: 19 euro). Workers in the clinical intervention group that had received usual care in the first 8 weeks returned to work 21.3 days (95% CI= [-74.1, 29.2]) later on average. The group that had received the workplace intervention in the first 8 weeks and the clinical intervention after 8 weeks returned to work 50.9 days (95% CI=[-89.4, -2.7]) later on average. A workplace intervention was more effective than usual care in RTW at slightly higher costs and was equally effective as usual care at equal costs on other outcomes. A clinical intervention was less effective than usual care and associated with higher costs. CONCLUSION: The workplace intervention results in a safe and faster RTW than usual care at reasonable costs for workers on sick-leave for two to six weeks due to LBP. PMID- 17086504 TI - The psychology of injured workers: health and cost of vocational rehabilitation. AB - BACKGROUND: In the vocational rehabilitation of injured workers the influence of the worker's personality has been neglected. This is despite there being substantial evidence that in chronic pain certain personality factors are significantly related to poorer outcomes. This is a preliminary study that has examined the relationship between personality factors, personality psychopathology and rehabilitation outcomes in injured workers. METHOD: Data from structured clinical interviews, self report and rehabilitation outcome (cost) were gathered from 36 injured workers with recognized compensation claims for physical and/or psychological workplace injury. RESULTS: Personality factors were associated with poorer outcome, particularly cost and health. Individuals with extreme personality traits experienced poorer health and vocational rehabilitation outcomes. The combination of high Neuroticism and low Extraversion which is a pattern often characterized as anxious and socially avoidant was found to be consistently related to poor health outcomes. CONCLUSIONS: The results indicate that considering the type of personality characteristics of injured workers may have important theoretical and practical implications. PMID- 17086505 TI - A parenteral econazole formulation using a novel micelle-to-liposome transfer method: in vitro characterization and tumor growth delay in a breast cancer xenograft model. AB - PURPOSE: The purpose of this study was to develop a parenteral liposomal formulation of econazole, a poorly water-soluble compound not previously available in an intravenous form. We are investigating econazole as an anticancer agent based on its unique mechanism of action to which cancer cells are preferentially sensitive. An intravenous formulation of econazole was desired for preclinical toxicity and efficacy studies of econazole. METHODS: Liposomal econazole was prepared using a novel micelle exchange technique to incorporate the drug into the lipid bilayer of pre-formed liposomes using a poly(ethylene) glycol-linked phospholipid, distearoyl phosphatidylethanolamine (DSPE-PEG). This method allowed for stable and efficient drug incorporation into DPPC and DMPC liposomes at a final drug:lipid ratio of 0.05 (w/w) and increased solubility in saline from <0.1 to 5 mg/ml. RESULTS: Stability over 14 days at 4 degrees C in buffer was demonstrated as well as in vitro plasma stability at 37 degrees C. Plasma elimination studies of micelle-loaded liposomal econazole showed a half life of approximately 35 min and plasma AUC of 281 microg/ml min. In MCF-7 human breast cancer xenografts in Rag2M mice. Liposomal econazole did not induce significant hepatoxicity, renal toxity or weight loss compared to empty liposomes. Tumor growth was slightly delayed in liposomal econazole-treated mice, with approximately 10-day lag time to reach 300 mm(3) compared to vehicle controls. CONCLUSIONS: The micelle transfer method provided an efficient means of preparing liposomal econazole suitable for intravenous administration. Liposomal econazole was successfully administered to tumor bearing mice at 50 mg/kg, and no significant toxicities attributable to econazole were observed. PMID- 17086506 TI - Cardiac resynchronization therapy in chronic heart failure. AB - Cardiac resynchronisation therapy (CRT) has emerged as a treatment option for patients with severe, drug-refractory heart failure and signs of intraventricular dyssynchrony. In clinical trials CRT reduced the overall mortality, improved symptoms, exercise tolerance, and left ventricular function, as compared with optimised medical therapy alone. One of the challenging fields in patient selection for CRT is to identify the 20-30% of heart failure patients with bundle branch block that will not respond to this novel therapy. Other fields of uncertainty, such as CRT in patients with atrial fibrillation or chronic right ventricular stimulation as well as the role of a back-up defibrillator will be discussed. PMID- 17086507 TI - Tissue engineered heart valves based on human cells. AB - Valvular heart disease is still a significant cause of morbidity and mortality worldwide. Clinically used valve replacements including mechanical valves as well as fixed biological xeno- or homografts are associated with several major disadvantages. Alternatively, tissue engineering aims at the fabrication of autologous living cardiovascular replacements with the potential to grow and to repair, particularly for paediatric applications. Therefore, autologous cells are harvested and seeded onto three-dimensional matrices followed by biomimetic in vitro conditioning enabling the development of the neo-heart valve tissue. Here, we review different human cell sources such as vessels, bone marrow, umbilical cord tissue and blood, and chorionic villi with particular regard to cell phenotypes and their suitability for extracellular matrix production for tissue engineering purposes. PMID- 17086508 TI - Invasive aspergillosis: is treatment with "inexpensive" amphotericin B cost saving if "expensive" voriconazole is only used on demand? AB - BACKGROUND: Voriconazole for the treatment of invasive aspergillosis (IA) shows superior clinical outcome and tolerability compared to conventional amphotericin B. However, the latter is often used as initial treatment due to lower drug acquisition costs. Therefore we performed a cost-effectiveness analysis. METHODS: A decision analytic model was designed to compare the cost-effectiveness of a regimen of voriconazole followed by conventional amphotericin B to a regimen of conventional amphotericin B followed by voriconazole. Patients initiated on treatment either completed initial therapy or switched to second line therapy due to toxicity or non-response. Probability of a switch was based on clinical trial data and local rates of renal toxicity. Resource use in the hospital was taken from the Global Comparative Aspergillosis (GCA) study. Costs were based on local drug acquisition costs, local cost estimates for hospitalisation and adjusted additional costs of amphotericin B-induced acute renal failure from the literature. Effectiveness was defined as survival at 12 weeks from the GCA study. An incremental cost-effectiveness ratio was estimated as the incremental cost per life saved comparing voriconazole to conventional amphotericin B. RESULTS: Based on this model, initial therapy of IA with voriconazole reduced total costs when compared to initial therapy with conventional amphotericin B (CHF 37 878/patient vs CHF 49 861/patient) and resulted in better survival at 12 weeks, making it the dominant treatment in terms of incremental cost-effectiveness. Results were most sensitive to alternative assumptions of the incidence of acute renal failure, but cost savings were sustained for voriconazole over a wide range of values. CONCLUSION: Considering that initial therapy with voriconazole is both cost saving and results in better clinical outcomes, voriconazole is the dominant cost effective option for initial therapy of IA, despite very low drug acquisition costs of conventional amphotericin B. PMID- 17086509 TI - Improvement of surgical skills after a three-day practical course for laparoscopic surgery. AB - OBJECTIVES: Currently, skills labs are becoming increasingly important in the field of medical education. This study aims to objectively assess psychomotor skills acquisition of residents attending a three-day laparoscopic course. MATERIALS AND METHODS: 44 participants (test group) of the sixth practical course for Visceral Surgery of German surgical societies (Deutsche Gesellschaft fur Chirurgie; DGCH and Berufsverband Deutscher Chirurgen; BDC) in Warnemuende with various degree of experience in laparoscopic surgery (18 advanced residents performing more than 50 laparoscopic operations and 26 novices performing less than 10 laparoscopic operations) and 6 consultants attending as tutors of the course (gold standard) were recruited as subjects. 20 medical students in their final year (camera holder) were chosen as a second control group (naive). Both control groups had no training during the practical course. The virtual reality simulator LapSim was used to assess laparoscopic skills of participants before and after the course. Time to complete the tasks, error score, and economy of motion parameters (path length and angular path) were analysed. RESULTS: After the practical course the advanced participants of the test group completed the task significantly faster (p = 0.019), with smaller error score (p = 0.023), and more economy of motion [path length (p = 0.014) and angular path (p = 0.049)] than before the course. The novices of the test group and both control groups showed no significant improvement of their performance parameters (p >0.05). CONCLUSION: A three-day practical course for laparoscopic surgery improved laparoscopic skills of residents. However, advanced residents benefit most from the course. PMID- 17086510 TI - Gender and the relationship between traumatic childhood experiences and pain in adulthood. AB - OBJECTIVES AND METHODS: Gender differences regarding 17 childhood experiences, thought to have traumatising potential (Traumatic Childhood Experiences = TCE), and pain behaviour in adulthood were assessed using a self-administered, anonymously filled-out questionnaire. Patients were consecutively accrued in the offices of practicing physicians. Three research questions were formulated: 1) Are specific TCE reported more frequently in male and female patients with the diagnosis "Pain Associated with Psychological Factors" (PP), compared to patients with "Pain, explained by Organic Processes" (OP), and "Patients with Diseases without Pain" (OD)? 2) Do PP-men and PP-women differ in reporting TCE?; 3) Are specific TCE correlated with Pain Duration, -Intensity and Number of Operations? RESULTS: 1). TCE occurred more frequently in PP-men and PP-women compared to OP- and OD-patients. 2). The PP-women reported much more TCE-items than the PP-men. 3). Duration and Intensity of adult pain associated with psychological factors correlated with certain TCE-items. CONCLUSIONS: The three research questions can be answered by "yes". In patients with pain which has been impossible to diagnose and/or has resisted conventional forms of therapy, TCE (verbal, physical and sexually abusive) have to be looked for, because they often explain adult pain. Unnecessary examinations and surgery can be avoided and therapies can be tailored for the individual patient. PMID- 17086511 TI - The influence of refractory ceramic fibres on pulmonary morphology, redox and immune system in rats. AB - Refractory ceramic fibres (RCF) were studied in male SPRD rats by both in vivo long term sequential and in vitro methods. RCF was administered by single intratracheal instillation and the lungs were examined at the end of months 1, 3 and 6 after exposure. In addition, the direct toxicity of the fibres was examined in a primary culture of alveolar macrophages (AM) and in pneumocytes type II (T2). Pulmonary morphological changes, a number of parameters of the redox system, such as activity of extracellular Cu,Zn/superoxide dismutase (EC-SOD), total glutathione content of the lungs (GSH) and immunoglobulins in bronchoalveolar lavage (IgA, IgG, IgM) and in the blood were measured. The composition of the original RCF and the elemental content of the lung tissue were compared by energy dispersive x-ray analysis (EDXA) before and after exposure. Macrophage alveolitis became confluent and moderate fibrosis developed by the end of month 3, and after 6 months of exposure the intensity decreased to the level of the first month. The RCF did not significantly influence the activity of EC SOD or the total glutathione content of the lungs. Although aluminium and silicon could be demonstrated by EDXA in the lung tissue at the end of month 3, these elements were no longer detectable by the end of month 6. The RCF decreased IgA significantly in bronchoalveolar lavage (BAL). The main components of RCF induced pulmonary alterations, whereas no significant change could be detected in EC-SOD and GSH. Injuries caused by direct toxicity could be observed in the cell membranes only at the highest concentration. On the basis of these results RCF can be determined as moderately toxic fibres. PMID- 17086512 TI - Characteristics of patients with rheumatoid arthritis presenting for physiotherapy management: a multicentre study. AB - OBJECTIVES: To describe the characteristics of patients with rheumatoid arthritis (RA) attending for physiotherapy management in Ireland. METHODS: Managers of physiotherapy departments in the 53 hospitals in Ireland were invited to participate in a multi-centre observational study over a 6-month period. Data on patients with RA the day of presentation for physiotherapy management were recorded. These data related to patient demographic details, disease management, aids and appliances, splint and orthoses usage and occupational issues. The Health Assessment Questionnaire was also recorded for each patient. RESULTS: A total of 273 patients from eight physiotherapy departments participated in the survey (n = 199; 73% female). Mean age of the participants was 59.3 (SD 12.5) years with mean disease duration of 13.8 (SD 10.6) years. The majority of the patients were inpatients (n = 170, 62%). Sixty-eight per cent of patients had attended for previous physiotherapy treatment and 98% were under current rheumatologist care. Biologic therapies were prescribed to 11% of patients. Use of splint and foot orthoses was high with 133 patients (49%) wearing splints and 75 (31%) wearing foot orthoses. The majority of patients had moderate (n = 119, 44%) or severe (n = 94, 35%) disability as per Health Assessment Questionnaire (HAQ) score. Mean HAQ score was 1.5, with HAQ scores showing increasing disability with increasing age, disease duration and erythrocyte sedimentation rate (ESR) levels. CONCLUSIONS: Patients with RA attending for physiotherapy management present with varied profiles. This study provides valuable information on the characteristics of patients with RA attending for physiotherapy management which will contribute to physiotherapy service planning and delivery and will optimize patient care. PMID- 17086513 TI - Family-based association tests for ordinal traits adjusting for covariates. AB - We present a class of family-based association tests (FBATs) for ordinal traits that adjust for the effects of covariates. For complex diseases, especially mental health conditions including nicotine dependence and substance use, the outcome variables are often recorded in an ordinal rather than quantitative scale. The naturally recorded ordinal traits are commonly analyzed either as quantitative traits or are dichotomized. It has been demonstrated repeatedly in recent studies that these commonly used approaches to dealing with ordinal traits are inadequate and result in loss of power. In this report, we make use of conditional likelihood to derive score test statistics that belong to a general class of FBATs. We conducted simulation studies to compare the type I error and power of our proposed test with existing tests. The empirical result suggests that our test produces reasonable type I errors and has power far exceeding (often doubling) those of existing tests. We applied our proposed test to a data set on alcohol dependence and found that six single nucleotide polymorphisms (SNPs) are significantly associated (P-values < or =0.001) with alcohol dependence after adjusting for gender and age. Three of the SNPs (rs619, rs1972373, and rs1571423) or their tightly linked regions have been suggested in the literature from the analysis of the same data, demonstrating the consistent findings between various methods. The other three SNPs (rs485874, rs718251, and rs1869907) are identified for the first time using this data set, underscoring the potential power of our proposed test. PMID- 17086514 TI - A new multimarker test for family-based association studies. AB - We propose a new multimarker test for family-based studies in candidate genes. We use simulations under different genetic models to assess the performance of competing testing strategies, characterized in this study as combinations of the following important factors: genes, statistical tests, tag single nucleotide polymorphisms (SNP) methods, number of tag SNPs and family designs. An ANOVA model is employed to provide descriptive summaries of the effects on power of the above-mentioned factors. We find that tag SNP methods, gene characteristics and family designs have minimal impact on the best testing strategy. The familywise error rate (FWER) controlling multiple comparison procedure and the new multimarker test offer the highest power followed by the asymptotic global haplotype test. Both the FWER and the multimarker test are invariant to family designs and gain power as we increase the number of tag SNPs. However, the performance of the global haplotype test is slightly degraded when analyzing larger numbers of tag SNPs. Within the framework of our study, the best strategy for family-based studies in candidate genes that emerged from our analysis is to use the FWER or the multimarker test and select 6-10 tag SNPs using any of the tag SNP methods considered. We confirm the conclusions of our study with an application to Alzheimer's disease data. PMID- 17086515 TI - The elusive goal of pedigree weights. AB - Non-parametric linkage analysis methods generally involve calculating an allele sharing statistic for each pedigree in a data set, then standardizing and summing the statistics over pedigrees. Pedigrees of different sizes can be weighted differently in the sum, though it is perhaps most common to weight all standardized pedigree statistics equally. Most other common weighting schemes are based on the number of affected individuals in the pedigree. It is also possible to derive optimal weights, which maximize power to detect linkage under particular trait models. We started by investigating three different analytical and simulation-based methods to calculate power and derive optimal weights. We found that simulation methods produce noticeably more accurate power calculations than the other methods. However, although the different calculation methods give different "optimal" weights, the power at those weights is very similar. That is, the analytical calculation methods are sufficient for finding good weights even though the simulation methods are most appropriate for calculating power. In comparing optimal weights for different trait models, we found that the weights vary quite a bit with the model, such that optimal weights for one model are not necessarily powerful at all for other models. Finally, we studied the power of a number of general weighting schemes, and of some new ones that incorporate information on how closely the affected individuals are related. We were able to find some schemes that performed well in the sense of giving reasonably powerful weights for most of the trait models and pedigree types we considered. PMID- 17086516 TI - Secret ties to industry and conflicting interests in cancer research. AB - BACKGROUND: Recently it was reported that a Swedish professor in environmental health has for decades worked as a consultant for Philip Morris without reporting his employment to his academic employer or declaring conflicts of interest in his research. The potential for distorting the epidemiological assessments of hazard and risk through paid consultants, pretending to be independent, is not exclusive to the tobacco industry. METHODS: Documentation is drawn from peer reviewed publications, websites, documents from the Environmental Protection Agency, University reports, Wellcome Library Special Collections and the Washington Post. RESULTS: Some consulting firms employ university researchers for industry work thereby disguising industry links in the income of large departments. If the industry affiliation is concealed by the scientist, biases from conflicting interests in risk assessments cannot be evaluated and dealt with properly. Furthermore, there is reason to suspect that editors and journal staff may suppress publication of scientific results that are adverse to industry owing to internal conflict of interest between editorial integrity and business needs. CONCLUSIONS: Examples of these problems from Sweden, UK, and USA are presented. The shortfalls cited in this article illustrate the need for improved transparency, regulations that will help curb abuses as well as instruments for control and enforcement against abuses. PMID- 17086517 TI - Proteolytic 18O-labeling strategies for quantitative proteomics. AB - A number of proteomic techniques have been developed to quantify proteins in biological systems. This review focuses on the quantitative proteomic technique known as "proteolytic 18O-labeling." This technique utilizes a protease and H(2)18O to produce labeled peptides, with subsequent chromatographic and mass spectrometric analysis to identify and quantify (relative) the proteins from which the peptides originated. The technique determines the ratio of individual protein's expression level between two samples relative to each other, and can be used to quantitatively examine protein expression (comparative proteomics) and post-translational modifications, and to study protein-protein interactions. The present review discusses various aspects of the 18O-labeling technique, including: its history, the advantages and disadvantages of the proteolytic 18O labeling technique compared to other techniques, enzymatic considerations, the problem of variable incorporation of 18O atoms into peptides with a discussion on recent advancements of the technique to overcome it, computational tools to interpret the data, and a review of the biological applications. PMID- 17086518 TI - Mechanical thrombectomy options in complex percutaneous coronary interventions. AB - Percutaneous coronary interventions (PCI) of thrombus-containing lesions are associated with an increased risk of acute complications and poorer long term vessel patency. Dealing with these vessels provides many technical challenges, especially with the significant risk of coronary no reflow and distal embolization. Pharmacological strategies, including intravenous and intracoronary glycoprotein IIbIIIa inhibitors reduce intracoronary thrombus propagation, improve TIMI flow and are associated with a reduction in adverse event rates. Mechanical strategies (particularly embolic protection and thrombectomy catheters) help to improve coronary blood flow and myocardial perfusion. However, their impact on clinical outcomes is less clear. The use of embolic protection devices is associated with better perfusion, blood flow, and clinical outcomes among patients undergoing saphenous vein graft (SVG) PCI. However, the role for these devices in primary PCI and native coronary artery interventions is uncertain. This study examines the current approaches to manage thrombotic lesions during PCI and reviews the evidence in support of the different mechanical thrombectomy options that are available to the interventional cardiologist. PMID- 17086519 TI - Transcatheter closure of perimembranous ventricular septal defects using umbrella devices. AB - There is only limited experience of interventional closure of perimembranous ventricular septal defects (pmVSDs), particularly on the long-term follow-up. This is a report on our long-term results after transcatheter closure of pmVSDs using the Cardioseal, Starflex, or Rashkind devices. PATIENTS: Between 1993 and 2005, we performed interventional occlusions of pmVSDs in 18 patients. The size of the defect ranged between 4 and 8.5 mm, Q(p)/Q(s) was calculated between 1.3 and 2.2. Except for two, the patients had no other structural heart defect. In the early days, we used the Rashkind PDA occluder (17 mm) in seven, followed by the Cardioseal device (17 mm) in nine, and the 23 mm Starflex device in two patients. RESULTS: Interventional closure of the defects was performed successfully in all patients without any complication during the procedure. Fluoroscopy times were 11.8-53.7 min (median 28.65 min). We achieved a complete closure in 13 patients, three patients with recently implanted devices still show minimal shunting. In two patients the occluder had to be removed surgically because of embolization into the pulmonary artery and significant residual shunting resulting in severe hemolysis in the second patient. In long- (mean 10.7 years) and short-term (mean 0.85 years) follow-up we have not observed any hemolysis, arrhythmias, device dislocations, or device-related aortic or tricuspid regurgitation. CONCLUSION: Transcatheter closure of small pmVSDs using non-selfcentering devices can be performed successfully. Long-term follow-up investigations show encouraging results. Complications like device-embolization and significant residual shunting occur in the presence of large defects and/or concomitant malformations. PMID- 17086520 TI - Successful device closure of an acquired Gerbode defect. AB - Left ventricle to right atrial communications, collectively known as Gerbode defects, are usually congenital defects and surgical closure remains the treatment of choice. We report a rare case of Gerbode defect acquired following surgical closure of a ventricular septal defect. The defect was successfully closed percutaneously with an Amplatzer ventricular septal occluder. The patient remained symptom free with optimal result on echo at 6 months of follow-up. This is the first reported case of successful device closure of an acquired Gerbode defect. PMID- 17086521 TI - Transcatheter closure of atrial septal defects in adults using two devices: an angiographic overview. AB - Atrial septal defects are a common congenital cardiac abnormality in adults. lndividuals with atrial septal defects carry the risk to develop cardiac problems such as right ventricle volume overload, pulmonary hypertension, atrial fibrillation, and more rarely paradoxical embolic events. Surgery was the standard treatment for symptomatic patients with atrial septal defects for decades. More recently, percutaneous transcatheter closure of atrial septal defects in adults using a closure device has become an alternative to surgery in selected patients. PMID- 17086522 TI - Cardiac catheterization in infants weighing less than 1,500 grams. AB - BACKGROUND: The improved survival of very low-birth-weight (<1,500 g) infants justifies more aggressive attempts to treat underlying congenital heart disease than in the past. METHODS: We retrospectively reviewed all catheterizations performed at our institution between January 1, 1990 and June 1, 2004 in infants weighing <1,500 g. We performed a 3:1 case-control study. Comparisons were randomly selected from a group of patients catheterized within 6 months of the cases and weighing 2-3 kg. All catheterization data, angiograms, and hospital charts were reviewed. RESULTS: Eighteen patients weighing <1,500 g underwent catheterization. Fifty-four patients were selected as comparisons. There were no significant differences in the age at catheterization, procedure time, fluoroscopy time, or contrast amount (cc/kg). The lower-birth-weight infants were more likely to be premature (median age 29 vs. 37 weeks, P < 0.001), and to have left-sided obstructive lesions including aortic stenosis or coarctation. The comparison patients were more likely to be postoperative (28% vs. 0%, P = 0.02), and included a higher number with hypoplastic left heart syndrome. There was an increased incidence of interventions performed in the lower-birth-weight infants (83% vs. 41%, P = 0.002). There was a difference in the interventions performed between the two groups: the comparisons had more atrial septal procedures, and the lower-birth-weight infants had more coarctation dilations and aortic valve dilations. There were no significant differences in the acute success rate of the procedures (100% vs. 95%), overall complication rate (56 vs. 57%), incidence of blood transfusions (44 vs. 30%), or major complications (11 vs. 13%) between the lower-birth-weight and comparison groups respectively. There was a trend towards higher survival rate in the comparison group in this small study population, but it did not reach significance (80 vs. 61%, P = 0.13). CONCLUSIONS: Cardiac catheterization in neonates <1,500 g is more likely to include percutaneous intervention, especially on the left side, but is generally successful with a complication rate similar to procedures performed in larger infants. Although these procedures are rare, improved miniaturization of equipment would facilitate safer interventions. PMID- 17086523 TI - Percutaneous PFO closure for the prevention of recurrent brain abscess. AB - A patent foramen ovale (PFO) can act as a conduit between the venous and arterial circulations, allowing right-to-left shunting and bypass of the pulmonary circulation. Brain abscess may develop as a result of paradoxical embolism of organisms through a PFO. In this small series, we report on the closure of PFO for the prevention of recurrent brain abscess. Only prospective, randomized trials comparing PFO closure to conservative therapy could provide a definitive answer as to the optimal strategy for preventing recurrent cerebral abscess. PMID- 17086524 TI - The "pull-push" technique to deal with a redundant eustachian valve interfering with placement of a PFO occluder. AB - OBJECTIVES: To evaluate the role of a new technique to deal with a redundant Eustachian valve (EV) interfering with placement of a patent foramen ovale (PFO) occluder. BACKGROUND: The EV is a remnant of the embryonic valve of the sinus venosus. In rare occasions, it is large and redundant and it may interfere with catheters and devices used in cardiac catheterization. METHODS: We have developed a new technique to control the EV in order to avoid interferences with the device for closure of atrial septal defects and PFO's. This technique is called "pull push" technique and consists in pulling down on the inferior vena cava the redundant EV by using a 6 Fr pigtail catheter while a second operator pushes the right atrial disc out of the long sheath. RESULTS: Here, we report on two patients with a patent foramen ovale, in whom the EV prevented the right atrial disc of an Amplatzer PFO occluder to be completely flat against the interatrial septum. Finally, we used this technique in one more patient to avoid any interference of a very redundant EV during placement of a Starflex device. CONCLUSIONS: In conclusion. the so called "pull-push" technique is an alternative and easy technique to control the EV in order to avoid interferences with the device for closure of atrial septal defects and PFO's PMID- 17086525 TI - Incidental findings with cardiac CT evaluation: should we read beyond the heart? AB - Cardiac computed tomographic angiography (CTA) allows for simultaneous evaluation of the lung fields and associated structures. There is a debate as to the benefit of or need for routine overread of the lung fields for incidental findings. The possible improvement in cancer diagnosis with routine overreads is balanced against the major limitations of CT lung screening. Current limitations include (a) a high rate of nodule detection given that >50% of participants may have at least one noncalcified nodule; (b) the increased costs and radiation exposure associated with the resulting follow-up CT scans; (c) the cost and the morbidity of follow-up, including further testing, as well as biopsy or resection of benign noncalcified nodule (at least 25% of such procedures in several trials); (d) a small but difficult to quantify potential risk of cancer associated with multiple follow-up CT scans; and (e) a potential for increased anxiety of both the patient and the physician about nonsignificant pathology. All of these limitations are balanced against a possibility that this could lead to an earlier detection of lung cancer with the consequent improvement in the chances of the patients' survival. Extensive studies of screening CT in older smokers have revealed the prevalence of cancer to be between 0.3 and 1%. However, when applied to an ambulatory population of patients presenting for an evaluation of angina, the prevalence of lung cancer or significant non-cardiac findings may be significantly lower. We have reviewed all the relevant literature and sought to determine the potential benefits and harms of specifically overreading CTA for non-cardiac pathology. The weight of the evidence suggests that it is most prudent to not specifically reconstruct and re-read CTA scans for lung nodules. If a non-cardiac abnormality is visualized by the primary interpreter of the cardiac CT, appropriate referral or follow-up is prudent. PMID- 17086526 TI - Mastering the antegrade femoral artery access in patients with symptomatic lower limb ischemia: learning curve, complications, and technical tips and tricks. AB - BACKGROUND: Antegrade femoral access is fraught by technical challenges and steeper learning curve, in comparison with retrograde contralateral femoral access. We appraised learning curve, complications, and technical aspects inherent in the adoption of antegrade approach. METHODS: Consecutive cases in which antegrade access was attempted by a cardiologist experienced in retrograde access, but inexperienced in antegrade, under supervision of an operator with anterograde expertise, were collected. The primary end-point was the occurrence of antegrade access failure or local complications. Major complications were defined as those life-threatening, requiring transfusion, percutaneous, or surgical repair. RESULTS: Anterograde access was attempted in 120 patients. The primary end-point occurred in 14 (11.6%) cases, but according to the learning curve, in 12 (20%) for first 60 cases vs 2 (3.3%) for the last 60 cases (P = 0.008). Access failure in the hands of the in-training operator was similarly found in all cases but one during the first 60 cases. No major complications occurred, while minor complications were found in 9 (7.5%) patients, again with all but two of them occurring in the first 60 cases. These included peri adventitial extravasation in 8 patients (6.7%), and perforation of a small branch in one (0.8%); all these complications were conservatively and successfully managed. Obesity was the only significant predictor of access failure/complication (P = 0.004). CONCLUSIONS: This work, the first to report on the learning curve of the antegrade approach, supports the feasibility and safety of this access site even for an in-training operator, if supervised. A minimum caseload of 60 procedures is likely needed to master this technique. PMID- 17086527 TI - Intravascular ultrasound identification of intraluminal embolic plaque material during carotid angioplasty with stenting. AB - INTRODUCTION: Carotid angioplasty with stenting (CAS) has evolved as a viable method for treating patients at high risk for carotid endarterectomy. Strokes complicating CAS are most commonly caused by the liberation and distal embolization of embolic material from plaque during the procedure. METHODS: CAS with distal embolic protection (DEP) was performed using the assistance of intravascular ultrasound (IVUS) in a 68-year-old man with asymptomatic carotid artery stenosis. Evaluation of the plaque pretreatment was performed using virtual histology software. IVUS evaluation was also performed after prestent angioplasty, stenting, and poststent angioplasty. RESULTS: Initial degree of angiographic stenosis was 78.9%. Final degree of stenosis was 15.7%. By IVUS, stenosis by minimum lumen diameter and minimum lumen area were 75.6% and 93.4% pretreatment, respectively, and 42.2% and 47.1% posttreatment, respectively. An intraluminal lesion was noted in the distal aspect of the stent after poststent angioplasty but before DEP device removal. Ultrasonographic characteristics of the intraluminal defect were consistent with ruptured plaque material. Angiographic runs failed to demonstrate the lesion. A repeat IVUS run performed approximately 10 min later failed to depict the lesion, suggesting that distal embolization had occurred. Embolic material was noticed in the DEP device after removal. The patient did not experience any ischemic neurological symptoms. CONCLUSIONS: IVUS can identify intraluminal plaque material, which may be a precursor for embolism and delayed ischemic events after CAS. IVUS may allow for treatment before distal embolization of plaque material. Virtual histology IVUS software may help to identify carotid lesions at higher risk for significant embolization during CAS. PMID- 17086528 TI - Mediastinal bronchial artery aneurysms: endovascular therapy in two patients. AB - We report two patients with cystic fibrosis and mediastinal bronchial artery aneurysms treated by means of endovascular coil embolization. In one case, the presence of a long segment of bronchial artery proximal to the aneurysm allowed coil embolization with regular steel coils. In the second case, the aneurysm was near the origin of the bronchial artery from the aorta. We used detachable coils to occlude this aneurysm for more precise embolization. PMID- 17086529 TI - Sirolimus-eluting stents and calcified coronary lesions: clinical outcomes of patients treated with and without rotational atherectomy. AB - This study examined the outcomes of patients who underwent sirolimus-eluting stent (SES) implantation for the treatment of heavily calcified coronary lesions (HCCL) with and without the use of rotational atherectomy (rotablator). We investigated 150 consecutive patients with angiographic evidence of HCCL who underwent SES implantation. Sixty-nine patients underwent SES implantation without the need of rotablator (SES), and 81 patients required rotational atherectomy to modify the plaque and facilitate the delivery of the stent (SES + rotational atherectomy). Clinical success was equivalent in both groups (>98%) and there were no in-hospital outcome differences. At 6 months, the target lesion revascularization rate was 4.9% in SES vs. 4.2% in SES + rotational atherectomy groups, respectively (P = NS). Mortality at 6 months was 7.9% in the SES group vs. 6.8% in the SES + rotational atherectomy group (P = NS). SES performs well in patients with complex HCCL, with a relative low event rate. Lesions requiring rotational atherectomy to facilitate dilation and stenting had similar outcomes after SES implantation to those that could be stented without the need for rotablator. PMID- 17086530 TI - Outcome of percutaneous coronary intervention in HIV-infected patients. AB - We report the long-term outcome of percutaneous coronary interventions (PCI) in 12 HIV-infected patients. Two patients died from a HIV-related infection and seven patients suffered from severe clinical and/or angiographic restenosis requiring additional interventions or causing severe angina pectoris. Only three patients remained symptom free. We conclude that HIV-infected patients should be considered as high risk group and treated routinely with drug-eluting stents. PMID- 17086531 TI - Percutaneous coronary intervention with stent deployment in anomalously-arising left circumflex coronary arteries. AB - BACKGROUND: Anomalous origin of the left circumflex coronary artery from the right sinus of Valsalva or proximal right coronary artery is the most common congenital coronary anatomical abnormality. Whether such vessels are particularly predisposed to atherosclerotic disease in their proximal portion remains controversial. Successful balloon angioplasty has been described, but thus far only six isolated cases of stent deployment in anomalous circumflex vessels have been described in the interventional literature. METHODS: Single-center retrospective case study of twenty-two patients with anomalous circumflex coronary arteries. Twelve patients (11/12 male, average age (63 +/- 3) years) underwent successful percutaneous coronary intervention (PCI) with stent deployment in the retroaortic segment of the anomalous vessel. RESULTS: Significant obstructive coronary disease in the retroaortic portion was found in the majority of anomalous left circumflex vessels (16/22, 73%), but did not predict the presence of significant two or three vessel coronary disease (P = 0.14, 0.63). PCI was successful in all cases attempted, with favourable short/medium-term event-free survival. CONCLUSIONS: The retroaortic portion of anomalously-arising circumflex coronary arteries is selectively predisposed to atherosclerotic disease. PCI to these vessels is feasible but may be technically challenging. PMID- 17086532 TI - Drug-eluting stents in bifurcation lesions: to stent one branch or both? AB - OBJECTIVES: The objective of this study was to compare two techniques to treat bifurcation lesions: a single drug-eluting stent (DES) implanted in the main branch combined with balloon dilatation for the side branch vs. stenting of both branches (double stent). BACKGROUND: Percutaneous coronary intervention in coronary bifurcation lesions remains challenging. Although DES reduce restenosis in lesions, the double stent procedure has not shown clear advantages over a single stent with balloon dilation. METHODS: Fifty-three symptomatic patients with true bifurcation lesions were treated using either the double stent technique (n = 25) or one stent in the parent vessel plus balloon angioplasty of the side branch (n = 28). Procedural results and major adverse cardiac event rates (MACE: cardiac death, myocardial infarction, target vessel revascularization (TVR)) were compared. RESULTS: Angiographic procedural success (residual stenosis <30% in both branches) was 75% in the single stent group and 100% in the double stent group (P = 0.01). All differences were due to residual stenosis of the side branch. Clinical follow-up (6-18 months) was available for all patients; 90.5% of patients had a coronary angiography or nuclear stress test. Three patients (11%) in the single stent group and two (8%) in the double stent group had ischemia-driven TVR (P = NS). Asymptomatic angiographic restenosis (>50% diameter stenosis) in the ostium of the side branch was seen in two patients in the double-stent group. At 6 months, MACE-free was comparable between groups (89.3% vs. 88%, P = 0.7). CONCLUSIONS: When treating bifurcation lesions with sirolimus-eluting stents, restenosis following a single stent procedure is comparable to stenting both parent and side branch vessels. Thus, stenting the main-branch lesion, coupled with balloon angioplasty in the side branch, produces a high success rate and good clinical outcomes at 6 months. PMID- 17086533 TI - Coronary air embolism: a case report and review of the literature. AB - Coronary air embolism is a complication in the catheterization laboratory that can be associated with high morbidity and even mortality. A case report of air embolism and methods to prevent this complication from occurring are presented along with various management techniques. PMID- 17086534 TI - A new technique to anchor stents for exact placement in ostial stenoses: the stent tail wire or Szabo technique. AB - This report describes a technique for correct positioning of a stent in an ostial stenosis by using a second wire passed through the last cell of a stent. The anchor wire technique, first described by Szabo et al. [Szabo S, Abramowitz B, Vaitkus PT. Am J Cardiol 2005;96:212H], will facilitate precise ostial stent placement and eliminate errors of positioning inside or outside the ostial narrowing. PMID- 17086535 TI - A new understanding of chronic total occlusion from a novel PCI technique that involves a retrograde approach to the right coronary artery via a septal branch and passing of the guidewire to a guiding catheter on the other side of the lesion. AB - Percutaneous coronary intervention (PCI) for chronic total occlusion (CTO) by the antegrade approach is sometimes difficult, especially in the right coronary artery (RCA). We performed successful PCls following a retrograde approach via a septal branch in 2 patients with CTO in RCA. The method involves leading the retrograde guidewire outside the body through an opposite guiding catheter after the wire crosses the target lesion. A balloon or stent could then be delivered retrogradely or antegradely. Even a soft retrograde wire always crosses the lesion through the true lumen, as confirmed by IVUS. Selecting a suitable collateral, a straighter rather than a larger one, is crucial. Our results do not support the current concept regarding CTOs. Probably, the distal fibrous cap is soft and the proximal one has a thin point that soft wires, even blunt ones, can penetrate easily. The distal penetration point appears to connect to the proximal uncalcified thin point. Many channels seem to spread out from the proximal side, tree-like, within the lesion. In the retrograde approach, the wire is unlikely to enter these branch channels. The results suggest that PCI by the retrograde approach may be effective for treating CTOs of RCA. PMID- 17086536 TI - Chronic total occlusion angioplasty through supplying collaterals. AB - Chronic total occlusions are typically difficult to recannalize especially when adverse angiographic morphologies are identified. We describe a case of chronic total occlusion crossing retrograde through the supplying collaterals followed by successful angioplasty and stenting. PMID- 17086537 TI - Transcatheter repair of iatrogenic left ventricular free-wall perforation. AB - Left ventricular free-wall perforation can complicate catheter-based diagnostic or interventional procedures and may require immediate needle pericardiocentesis followed by surgical repair in about 20% of the cases. We describe the transcatheter closure of a left ventricular free-wall perforation as an option in the event of maintained access to the perforation site after defect creation. PMID- 17086538 TI - B-type natriuretic peptide level in a patient with constrictive pericarditis. AB - We report the case of a 35-year-old man with constrictive pericarditis who had a B-type natriuretic peptide (BNP) level of 129 pg/dl despite a left ventricular end diastolic pressure of 35 mmHg. We discuss a possible explanation for the relatively low BNP level given this patient's markedly elevated intracavitary pressures in the setting of constrictive pericarditis. PMID- 17086539 TI - Axillary artery approach for balloon valvoplasty in young infants with severe aortic valve stenosis: medium-term results. AB - OBJECTIVES: To assess the feasibility and results of trans-axillary approach for balloon aortic valvoplasty (BAV) in early infancy. BACKGROUND: Severe aortic valve stenosis (SAVS) is rare but serious condition in infancy, which may be promptly treated either by surgical aortic valvotomy or BAV. BAV is usually performed via the femoral artery route, which is associated with significant vascular complications and long procedure times. METHODS: BAV via the trans axillary approach was performed on twenty-seven sequential infants with SAVS presenting to a single tertiary referral center over an 11-year period. Maximum inflated balloon size was less than or equal to the aortic valve diameter. RESULTS: Twenty-seven infants aged 1-77 days underwent BAV. Weight at time of procedure was 2.0-4.42 kgs. The median procedure and screening times were 82 and 7.9 minutes, respectively. Mean instantaneous Doppler gradient across the aortic valve reduced from 68 +/- 33 to 37 +/- 14 mmHg ( p < 0.0001). Three infants developed at least moderate aortic regurgitation. Right arm pulse volume was decreased in 12 infants; 5 received an intravenous heparin infusion. Longer-term follow-up demonstrated reduced or absent peripheral pulse in 5 infants. Transection of the axillary artery occurred in one infant requiring emergency microvascular repair. There was one post-procedural and one late death due to non cardiac causes. CONCLUSIONS: In early infancy balloon aortic valvoplasty via the axillary artery approach for severe aortic stenosis is an acceptable and safe alternative to the femoral arterial approach and results in short procedure and screening times. Longer-term vascular follow-up is required. (c) 2006 Wiley-Liss, Inc. PMID- 17086540 TI - Dysfunctional attitudes and personality disorder comorbidity during long-term treatment of MDD. AB - A key component of how depression may impact personality pathology involves an understanding of how cognition and dysfunctional attitudes may change as a result of experiencing a depressive state, and how these changes may affect reporting of personality disorder symptoms. This study examines whether dysfunctional attitudes are related to the stability of personality disorder diagnoses. The sample comprised 64 outpatients who were treatment responders following an 8-week acute treatment phase for major depressive disorder (MDD), met criteria for remission throughout a 26-week continuation phase, and completed a personality disorder assessment Structured Clinical Interview for DSM-III-R Axis II Disorders (SCID-II) at the beginning and end of each treatment phase. The Dysfunctional Attitude Scale (DAS) was given to patients at the beginning of the continuation phase. We found that following successful treatment of the MDD, individuals with stable personality disorder diagnoses (e.g., meeting criteria for a personality disorder at both the beginning and endpoint of continuation treatment) had greater severity of dysfunctional attitudes (P =.001) at the beginning of the continuation treatment compared to those who never met criteria for a personality disorder during continuation treatment. Though there was no significant relationship between DAS scores and the stability of a Cluster A or Cluster B personality disorder diagnosis, there was a significant relationship between DAS scores and the stability of a Cluster C personality disorder diagnosis (P <.001). Outpatients who had a stable Cluster C personality disorder diagnosis had higher scores on the DAS at the beginning of continuation treatment compared to outpatients who never met criteria for a Cluster C diagnosis. This finding suggests that dysfunctional attitudes that persist beyond remission of MDD may be a marker for certain personality disorders that are stable across long-term treatment. PMID- 17086541 TI - Direct aspiration using rapid-exchange and low-profile device for acute thrombo embolic occlusion of the superior mesenteric artery. AB - Acute thrombo-embolic occlusion of the superior mesenteric artery (SMA) is potentially fatal vascular emergency that requires rapid restoration of mesenteric blood flow as well as early diagnosis. Selective thrombolysis has been employed for this life-threatening event. Although failed thrombolysis is a well known phenomenon, which is directly connected with bowel necrosis, emergent laparotomy, and patient death, little progress has been made in its treatment strategy. Recently, direct aspiration using rapid-exchange and low-profile aspiration device has been reported as a simple, safe, and effective strategy to thrombus in coronary artery. We describe the utility of direct aspiration using this sort of aspiration device for failed thrombolysis of SMA thrombo-embolism. PMID- 17086542 TI - Dynamic association with polysomes during P19 neuronal differentiation and an untranslated-region-dependent translation regulation of the tau mRNA by the tau mRNA-associated proteins IMP1, HuD, and G3BP1. AB - Regulation of mRNA translation is a key step in mediating neuronal polarity during differentiation, insofar as neuronal polarity is partially determined by local translation of specific mRNA molecules as dendrites and axons are emanating. The multiplicity of mRNA-binding proteins in neurons plays an essential role in controlling mRNA translation. These proteins are associated with ribosomes and translation factors, thereby regulating both temporally and spatially the translation process. In a previous study, we have shown an association among the tau mRNA-binding proteins HuD, IMP1, and G3BP1 with translating polysomes in P19 neurons. In the present study, we determined the dynamics of the association among G3BP1, IMP1, and HuD with polysomes through P19 neuronal differentiation as well as the functional effect of these proteins on tau mRNA translation. We show a novel, differentiation-dependent association of these proteins with polysomes. In addition, we show a strong, negative effect on translation of the tau mRNA by IMP1, G3BP1, and HuD proteins in HEK-293 cells. To our knowledge this is the first observation of a direct translational role of G3BP1 for any mRNA and the first report of a translation inhibition by IMP1 and HuD on the tau mRNA in a cell system. The translation inhibition is shown to be mediated by the tau mRNA 3'untranslated regions (UTRs), thus giving a new, translational role for these sequences, which were previously implicated in mRNA stabilization. We also define a novel mechanism for IMP1 binding to tau mRNA, which suggests a conformational binding, which is not sequence dependent. PMID- 17086543 TI - Febrile seizures are associated with mutation of seizure-related (SEZ) 6, a brain specific gene. AB - Genetic factors contribute significantly to the etiology of febrile seizures (FS), the most common type of seizures in childhood. However, in most patients with FS, the causative gene is unknown. The purpose of this study was to explore the relationship between human brain-specific gene SEZ-6 and FS. Through amplification of genomic DNA by PCR and sequencing of the resulting products, we screened 75 subjects for mutations in the coding region (17 exons) of the SEZ-6 gene. Fifteen subjects were healthy individuals and 60 subjects had FS. Patients with FS could be divided into sub-groups based on seizure type (42 simple and 18 complex) and family history (41 had a positive family history). All patients have been followed to date to evaluate seizure recurrence and the development of epilepsy. No mutations were found in healthy controls, but 21 of the patients with FS had mutations in SEZ-6, and the most common type of mutation was a heterozygous, cytosine insertion (frame shift mutation) at position 1435 of the cDNA. The mutation incidence was significantly higher in patients with complex FS (vs. simple FS) and in patients with a positive family history. Sixteen of 42 patients with simple FS experienced seizure recurrence during the 1-5-year follow up period. Fifteen of 18 patients with complex FS also experienced a recurrence during this period. Among these patients with recurrences, five patients with simple FS and six patients with complex FS have developed epilepsy. The mutation incidence among these epileptic patients is 72.7%. The human SEZ-6 gene is related to the occurrence and development of FS and may be a novel candidate gene for epilepsy. Screening for mutations in SEZ-6 may be valuable in predicting FS recurrence or the development of epilepsy. PMID- 17086544 TI - Expression, activity, and role of serine palmitoyltransferase in the rat hippocampus after kainate injury. AB - An increase in ceramide species has been shown recently by lipidomic analysis of the rat hippocampus after kainate-induced excitotoxic injury (Guan et al. [2006] FASEB J 20:1152-1161). In this study, we showed increased expression of serine palmitoyltransferase (SPT), the first enzyme in the ceramide biosynthetic pathway, in reactive astrocytes of the hippocampus after kainate injections. The increase in enzyme expression was paralleled by increased SPT enzyme activity in the hippocampus at 2 weeks post-kainate injection. In vitro studies showed that treatment of hippocampal slice cultures with SPT inhibitor ISP-1 (myriocin) or L cycloserine modulated increases in 16:0, 18:0, and 20:0 ceramide species, and partially reduced kainate-induced cell death. The above findings indicate a role of SPT in ceramide increase after kainate injury, although additional effects of sphingomyelinase cannot be ruled out. They also suggest that SPT activity might contribute to neuronal injury after kainate excitotoxicity. PMID- 17086545 TI - Inhibitors of glutamate transport modulate distinct patterns in brain metabolism. AB - High affinity uptake of glutamate plays a major role in the termination of excitatory neurotransmission. Identification of the ramifications of transporter function is essential to understand the diseases in which defective excitatory amino acid transporters (EAAT) have been implicated. In this work we incubated Guinea pig cortical tissue slices with [3-(13)C]pyruvate and major currently available glutamate uptake inhibitors and studied the resultant metabolic sequelae by (13)C and (1)H NMR spectroscopy using a multivariate statistical approach. Perturbation of glutamate uptake produced significant effects on metabolic flux through the Krebs cycle, and on glutamate/glutamine cycling rates, with this effect accounting for 76% of the variation in the total data set. The effects of all inhibitors were separable from each other along three major principal components. The competitive inhibitor L-CCG III ((2S,1'S,2'R)-2 carboxycyclopropyl)glycine) differed most from the other inhibitors, showing negative weightings on both the first and second principal components, whereas the EAAT2-specific inhibitor dihydrokainate (DHK) showed metabolic patterns similar to that of anti-endo-3,4-methanopyrolidine dicarboxylate but separate from those of DL-threo-beta-benzyloxyaspartate (TBOA) and L-trans-pyrrolidine-2,4 dicarboxylate (L-tPDC). This indicates that different inhibition mechanisms or different colocalisation of the separate transporter subtypes with glutamate receptors can produce significantly different metabolic and functional outcomes for the brain. PMID- 17086546 TI - Asynchronic transmission in the CA3-CA1 hippocampal synapses in the neurological mutant taiep rat. AB - For the taiep rat, a neurological mutant with severe astrogliosis secondary to demyelination, we have described alterations in spinal cord synaptic transmission. Asynchronous responses result from phasic action potential-derived glutamate release in this mutant. To evaluate whether this anomalous transmission is also produced in other regions of the taiep CNS and whether its nature involves a presynaptic or postsynaptic disruption, we studied the CA3-CA1 hippocampal synapses. Excitatory postsynaptic currents (EPSC) evoked by stimulation of Schaffer collaterals were recorded from CA1 pyramidal cells on picrotoxin-treated slices. Initial fast and time-locked EPSCs were evoked by conventional stimulation in both control and taiep neurons, showing similar latency and amplitude values unimodally distributed. In a high percentage of taiep neurons (47%), the initial EPSC was frequently followed by additional asynchronous synaptic currents (EPSC(ASYN)) with latencies ranging from 10 to 300 msec. As with initial EPSCs, EPSC(ASYN) were action potential dependent, sensitive to tetrodotoxin, and blocked by D-2-amino-5-phosphonovaleric acid and 6 cyano-7-nitroquinoxaline-2,3-dione. The occurrence probability of these events decayed monoexponentially as a function of poststimulus time. The elevation of extracellular Ca(2+) induced a reduction of amplitudes and a rate increase of EPSC(ASYN), in parallel with a reduction of paired pulse facilitation of initial EPSCs. The presynaptic fiber volley, extracellularly recorded, showed no significant differences between groups, with similar mean values of area and decay time. These findings in hippocampal circuitry suggest that, in taiep, the asynchronous evoked activity represents a rather generalized phenotype of the glutamatergic synapses and that EPSC(ASYN) seems to be determined by presynaptic alterations. PMID- 17086547 TI - Intracerebroventricular passive immunization with anti-oligoAbeta antibody in TgCRND8. AB - Based on the central dogma of beta-amyloid (Abeta) as a key seeding event in the pathogenesis of Alzheimer disease (AD), immunoneutralization strategies have been actively pursued both in AD and in models of AD as a potential means for treating AD. Both active and passive immunizations targeted at fibrillar Abeta successfully remove cerebral plaque load and attenuate Abeta-induced toxicity. Consistently with this, intracerebroventricular (ICV) passive immunization established in our laboratory using antibody against fibrillar Abeta (anti fAbeta) reduced cerebral plaque load and reversed early synaptic deficits at pre/early plaque stage when there is an abundance of soluble dimeric/oligomeric Abeta but sparse fibrillar Abeta, indicating that anti-fAbeta-mediated partial neutralization of toxic oligomeric Abeta species might have reduced early synaptotoxicity. In the previous investigation, we found that immunoneutralization with anti-fAbeta transiently reduced cerebral Abeta and associated toxicity. The current investigation tested whether ICV im munization using antibody to conformationally changed oligomeric Abeta (anti-oligoAbeta) will overcome the transient restorative nature of anti-fAbeta and produce persistent, long-lasting preventive effects. Because oligomeric Abeta is strongly correlated with synaptotoxicity, we investigated whether immunoneutralization of oligomeric Abeta will reverse synaptic deficits by analyzing presynaptic molecular marker (SNAP-25) profile within hippocampal dendritic fields, where SNAP-25 is abundantly expressed. Results show that, in contrast to ICV anti fAbeta antibody, ICV anti-oligoAbeta antibody significantly prevented cerebral Abeta build and almost completely restored SNAP-25 immunoreaction up to 8 weeks postinjection in TgCRND8 brain. Results show that ICV passive immunization with anti-oligoAbeta antibody might be an improved ICV immunization strategy for preventing permanent structural damage in AD. PMID- 17086548 TI - Role of brain-derived neurotrophic factor in the protective action of N-methyl-D aspartate in the apoptotic death of cerebellar granule neurons induced by low potassium. AB - Several neurotrophic factors, including brain-derived neurotrophic factor (BDNF), and neurotransmitters, such as glutamate, may influence neuronal apoptotic death. Rat cerebellar granule neurons (CGN) cultured in low potassium (5 or 10 mM KCl) for more than 5 days in vitro (DIV) die apoptotically. These cells survive in the presence of high potassium (25 mM KCl, K25) or N-methyl-D-aspartate (NMDA), an agonist of glutamatergic receptors. CGN transferred from high to low potassium die apoptotically. Here, we characterized the effect of BDNF and NMDA on the apoptotic death induced by low potassium in CGN. Cell death of CGN by culturing in low potassium for 6 DIV was inhibited by BDNF and NMDA. When CGN were cultured in K25 and transferred to a low-potassium medium, 65% of neurons died after 48 hr. Under these conditions, BDNF, NMDA, or BDNF + NMDA increased CGN survival. Both BDNF and NMDA decreased caspase-9 activity and mRNA caspase-3 levels and activity induced by low potassium. CGN survival induced by BDNF is mediated by TrkB activation, whereas that induced by NMDA is mediated by NMDA receptor and TrkB activation. NMDA, but not BDNF, raised [Ca(2+)](i), which was reduced by low potassium treatment. These results suggest that NMDA receptor stimulation induces CGN survival through the influx of extracellular Ca(2+) that may evoke the release of BDNF and the activation of TrkB. Complementary mechanisms induced by depolarization and changes in Ca(2+) levels would also contribute to the neuroprotection exerted by NMDA and potassium. PMID- 17086549 TI - Three-dimensional confocal morphometry reveals structural changes in astrocyte morphology in situ. AB - Neuronal activity and many pathological states in the CNS are accompanied by transient astrocytic swelling, which affects excitability, extrasynaptic transmission, and neuron-glia interactions. By using three-dimensional confocal morphometry (3DCM), we quantified the morphometric parameters of astrocytes in intact tissue. In experiments performed in brain cortex slices from transgenic GFAP/EGFP mice, we applied 3DCM to study the dynamic changes in astrocyte morphology during hypotonic stress. Our morphometric analysis showed that the effect of a 10-min application of hypotonic solution (200 mmol/kg) on the swelling of different cell compartments was dependent on the extent of the swelling of the total astrocyte volume. If the swelling of the whole cell, i.e., soma and processes, was less than approximately 10%, there were no differences between the swelling of the soma and the processes. However, if the swelling of the total cell volume was greater than 10%, the swelling of the processes was greater than the swelling of the soma. Analyzing the effect of hypotonic solution on the morphology of these astrocytes revealed that the total cell volume increased; however, certain cell compartments were distinguished in which the volume increased, whereas in other compartments cell volume decreased or apparently did not change, and the structure of some compartments was altered. Our data show that astrocytes in brain slices undergoing hypotonic stress display cell volume regulation as well as transient changes in morphology. PMID- 17086550 TI - Chronic but not acute intracerebroventricular administration of amyloid beta peptide(25-35) decreases somatostatin content, adenylate cyclase activity, somatostatin-induced inhibition of adenylate cyclase activity, and adenylate cyclase I levels in the rat hippocampus. AB - Although alterations in adenylate cyclase (AC) activity and somatostatin (SRIF) receptor density have been reported in Alzheimer's disease, the effects of amyloid beta-peptide (Abeta) on these parameters in the hippocampus are unknown. Our aim was to investigate whether the peptide fragment Abeta(25-35) can affect the somatostatinergic system in the rat hippocampus. Hence, Abeta(25-35) was injected intracerebroventricularly (i.c.v.) to Wistar rats in a single dose or infused via an osmotic minipump connected to a cannula implanted in the right lateral ventricle during 14 days. The animals were decapitated 7 or 14 days after the single injection and 14 days after chronic infusion of the peptide. Chronic i.c.v. infusion of Abeta(25-35) decreased SRIF-like immunoreactive content without modifying the SRIF receptor density, SRIF receptor expression, or the Gialpha(1), Gialpha(2), and Gialpha(3) protein levels in the hippocampus. This treatment, however, caused a decrease in basal and forskolin-stimulated AC activity as well as in the capacity of SRIF to inhibit AC activity. Furthermore, the protein levels of the neural-specific AC type I were significantly decreased in the hippocampus of the treated rats, whereas an increase in the levels of AC V/VI was found, with no alterations in type VIII AC. A single i.c.v. dose of Abeta(25-35) exerted no effect on SRIF content or SRIF receptors but induced a slight decrease in forskolin-stimulated AC activity and its inhibition by SRIF. Because chronic Abeta(25-35) infusion impairs learning and memory whereas SRIF facilitates these functions, the alterations described here might be physiologically important given the decreased cognitive behavior previously reported in Abeta-treated rats. PMID- 17086551 TI - Changes in the composition of detergent-resistant membrane domains of cultured neurons following protein kinase C activation. AB - Changes in the composition of cell fractions, and in particular of detergent resistant membranes (DRM) isolated from cultured rat cerebellar granule cells, were taken as possible changes in lipid raft composition during a signal transduction event. After activation of protein kinase C (PKC) with phorbol esters (PMA) or glutamate, the content of PKC and of proteins highly enriched (GAP43, Fyn, and PrP(c)) or not (MARCKS) in DRM was followed. PKC activation strongly increased its association with membranes (from 2% to 75%), causing its enrichment within DRM; the substrate GAP43, enriched in DRM, remained membrane associated, but its proportion in DRM dramatically decreased (from about 40% to 2.5%), suggesting its shift from raft to nonraft membranes, possibly as a consequence of phosphorylation by PKC. The distribution of Fyn and PrP(c) (DRM enriched) and of MARCKS (present mainly outside DRM) did not change. PKC activation was followed by an increase of GAP43 and MARCKS phosphorylation (about 7- and 8-fold, respectively). Noteworthy was that, after cell treatment with the lipid raft-disrupting drug methyl-beta-cyclodextrin, PKC activation occurred normally, followed by MARCKS phosphorylation, but GAP43 phosphorylation did not occur. Taken altogether, these data suggest that the integrity of lipid rafts is necessary for PKC to affect GAP43 and catalyze its phosphorylation. PMID- 17086552 TI - Cytomorphological diagnosis of hemangiopericytoma. PMID- 17086553 TI - Dramatic enhancement of enone epoxidation rates in nonionic microemulsions. AB - The ability of microemulsions to dissolve polar and non-polar components with a huge internal interface can overcome the reagent incompatibilities frequently encountered in organic reactions. We investigated model epoxidation reactions of alpha,beta-unsaturated enones and alkaline hydrogen peroxide in different nonionic microemulsions, both in the presence and absence of a phase-transfer agent (PTA). The obtained reaction profiles were compared with those for the corresponding surfactant-free two-phase systems. In addition, we defined a time constant tau as a measure for the rate of turnover. The epoxidation of trans chalcone using an n-alkyl-polyoxyethylene surfactant based microemulsion was fastest in the system with the PTA (tau=66 min) and slightly slower without the PTA (tau=77 min). It was still slower in the two-phase system with a PTA (tau=114 min) and extremely sluggish without a phase-transfer agent. With n-alkyl beta-D glucopyranoside as the surfactant the conversion was twice as fast than in the former microemulsion systems, but the PTA did not accelerate the reaction further (tau=35 and 33 min). The epoxidation of vitamin K(3), the second model system, was extremely accelerated. It proceeded a factor of approximately 35 faster in the microemulsion (tau=1.44 min) than in the corresponding two-phase system (tau=57 min). PMID- 17086554 TI - WIP null mice display a progressive immunological disorder that resembles Wiskott Aldrich syndrome. AB - The Wiskott-Aldrich syndrome (WAS) is an X-linked immunodeficiency syndrome caused by mutations in the WAS protein (WASP). This participates in signalling and cytoskeletal homoeostasis, and some of its activities are regulated by its binding to the WASP interacting protein (WIP). WIP deficiency, however, has not yet been shown to be of pathological significance in humans. Here we show that, in WIP null (WIP(-/-)) mice, it produces haematological alterations and anatomical abnormalities in several organs, most probably as a consequence of autoimmune attacks. Granulocytosis and severe lymphopenia are associated with a proportional increase in segmented cells and fewer bone marrow erythrocytes and lymphocytes. Splenomegaly is accompanied by an increase of haematopoietic tissue and red pulp, reduction of the white pulp, and fewer B (B220(+)) lymphocytes (also apparent in the lymph nodes and Peyer's patches). Ulcerative colitis, interstitial pneumonitis, glomerular nephropathy with IgA deposits, autoantibodies, and joint inflammation are also evident. These progressive immunological disorders closely mimic those seen in WAS. WIP deficiency may thus be implicated in some cases in which mutations in the gene encoding WASP are not detected. PMID- 17086555 TI - The course of severe fatigue in disease-free breast cancer patients: a longitudinal study. AB - BACKGROUND: We investigated whether fatigue is a persistent problem, and whether persistent fatigue is related to former treatment modalities. In addition, we studied the predictors of persistent fatigue. METHODS: At baseline (n=150, mean time since cancer treatment=29 months) patients were asked to fill out several questionnaires on psychological, physical, social, cognitive and behavioral aspects (Ann. Oncol. 2002;13:589-598). During the 2 years after baseline patients were asked to fill out monthly a fatigue questionnaire (CIS-fatigue). Hundred twenty-one patients completed the study, 10 dropped out and 19 had a disease recurrence. RESULTS: Twenty-four percent of the patients experienced persistent severe fatigue complaints during the 2-year observation period. Persistent fatigue seemed to be related to the duration of former treatment but unrelated to type of surgery, type of adjuvant therapy and time since treatment finished. High anxiety, high impairment in role functioning and low sense of control over fatigue symptoms at baseline were predictors of persistent fatigue. CONCLUSION: Fatigue appears to be a persistent problem for a quarter of a sample of disease free breast cancer patients during a 2-year period. The predictors of persistent fatigue found in this study can be helpful for the development of interventions to reduce post-treatment fatigue. PMID- 17086556 TI - An amine-promoted aziridination of chalcones. PMID- 17086557 TI - Silver chalcogenide clusters with dimethylanilinomercapto ligands: syntheses and crystal structures of [Ag65S13(SC6H4NMe2)39(dppm)5], [Ag76Se13(SC6H4NMe2)50(PPh3)6.5], and [Ag88Se12(SC6H4NMe2)63(PPh3)6]. PMID- 17086558 TI - Regulation of exocytosis in chromaffin cells by trans-insertion of lysophosphatidylcholine and arachidonic acid into the outer leaflet of the cell membrane. AB - Vesicular exocytosis is an important complex process in the communication between cells in organisms. It controls the release of chemical and biochemical messengers stored in an emitting cell. In this report, exocytosis is studied amperometrically (at carbon fiber ultramicroelectrodes) at adrenal chromaffin cells, which release catecholamines after appropriate stimulation, while testing the effects due to trans-insertion of two exogenous compounds (lysophosphatidylcholine (LPC) and arachidonic acid (AA)) on the kinetics of exocytotic events. Amperometric analyses showed that, under the present conditions (short incubation times and micromolar LPC or AA solutions), LPC favors catecholamine release (rate, event frequency, charge released) while AA disfavors the exocytotic processes. The observed kinetic features are rationalized quantitatively by considering a stalk model, for the fusion pore formation, and the physical constraints applied to the cell membrane by the presence of small fractions of LPC and AA diluted in its external leaflet (trans insertion). We also observed that the detected amount of neurotransmitters in the presence of LPC was larger than under control conditions, while the opposite trend is observed with AA. PMID- 17086559 TI - Glycine conjugates in a lepidopteran insect herbivore--the metabolism of benzylglucosinolate in the cabbage white butterfly, Pieris rapae. AB - Herbivores have developed a wide array of countermeasures to overcome plants' chemical defences. Larvae of the cabbage white butterfly, Pieris rapae, feed exclusively on plants of the Brassicales order, which are defended by the glucosinolate-myrosinase system. The defensive function of this system comes from toxic isothiocyanates that are formed when glucosinolates are hydrolysed by myrosinases upon tissue damage. Here we show that P. rapae larvae convert benzylglucosinolate to phenylacetylglycine, which is released in their faeces. Feeding experiments with isotopic tracers suggest that phenylacetonitrile and phenylacetic acid are intermediates in this conversion. We also identified additional glycine and isoserine (2-hydroxy-3-aminopropanoic acid) conjugates with benzoate and indole-3-carboxylate from P. rapae faeces extracts. This is the first description of such conjugates from lepidopteran insects. PMID- 17086560 TI - Catalytic relationships between type I and type II iterative polyketide synthases: The Aspergillus parasiticus norsolorinic acid synthase. AB - Norsolorinic acid synthase (NSAS) is a type I iterative polyketide synthase that occurs in the filamentous fungus Aspergillus parasiticus. PCR was used to clone fragments of NSAS corresponding to the acyl carrier protein (ACP), acyl transferase (AT) and beta-ketoacyl-ACP synthase (KS) catalytic domains. Expression of these gene fragments in Escherichia coli led to the production of soluble ACP and AT proteins. Coexpression of ACP with E. coli holo-ACP synthase (ACPS) let to production of NSAS holo-ACP, which could also be formed in vitro by using Streptomyces coelicolor ACPS. Analysis by mass spectrometry showed that, as with other type I carrier proteins, self-malonylation is not observed in the presence of malonyl CoA alone. However, the NSAS holo-ACP serves as substrate for S. coelicolor MCAT, S. coelicolor actinorhodin holo-ACP and NSAS AT domain catalysed malonate transfer from malonyl CoA. The AT domain could transfer malonate from malonyl CoA to NSAS holo-ACP, but not hexanoate or acetate from either the cognate CoA or FAS ACP species to NSAS holo-ACP. The NSAS holo-ACP was also active in actinorhodin minimal PKS assays, but only in the presence of exogenous malonyl transferases. PMID- 17086561 TI - A protein fluorescence amplifier: continuous fluorometric assay for rab geranylgeranyltransferase. PMID- 17086562 TI - Manipulation of the morphology of semiconductor-based nanostructures from core shell nanoparticles to nanocables: the case of CdSe/SiO(2). AB - The morphology of CdSe/SiO(2) was manipulated from core-shell-structured nanoparticles to nanocables by using a chemical vapor deposition (CVD) process. The growth of nanocables, with cores no more than 20 nm in diameter, is initiated by the formation of core-shell nanoparticles with SiO(2) as matrix and CdSe clusters dispersed inside. After the subsequent vaporization of the SiO(2) matrix, the follow-up CdSe vapor crystallizes with the remaining CdSe clusters as nuclei to form CdSe nanowires as the furnace was cooled to 1200 degrees C. During the controlled cooling of the furnace, the SiO vapor re-deposits to sheathe the nanowires. The thickness of the shell and the diameter of core were successfully controlled. The photoluminescence measurements show that the CdSe/SiO(2) nanocables have strong visible-light emission bands located at 590 and 688 nm, which are attributed to the defects induced by SiO(2) sheaths nanowires and the quantum confinement effect of the CdSe, respectively. The UV/Vis absorption spectra of the naked CdSe nanowires further validate the above-mentioned quantum confinement effect. The deterministic growth of these nanocables is very important for the design of the nanodevices based on them. PMID- 17086563 TI - Use of cyclo-oxygenase 2 inhibitors (COX-2) and prescription non-steroidal anti inflammatory drugs (NSAIDS) in UK and USA populations. Implications for COX-2 cardiovascular profile. AB - BACKGROUND: COX-2 and NSAIDS differ in their gastrointestinal (GI) and cardiovascular (CV) toxicity from pharmacological, clinical and epidemiologic point of views. OBJECTIVE: Describe the patterns of use of NSAIDS and COX-2 in The Health Improvement Network (THIN) database in UK and the PharMetrics database in USA. METHODS: We examined the experience of 10 distinct cohorts of new users of diclofenac, naproxen, ibuprofen, piroxicam, other NSAIDS, meloxicam, celecoxib, etoricoxib, rofecoxib and valdecoxib. The study period was 1 January 1995 through 2004 (31 March in UK and 28 February in USA). We collected information on covariates including history of upper GI disease, CV disease, hepatic disease, dosage, concomitant medication, and visits to a rheumatologist. RESULTS: We identified 486 076 unique patient-drug pairs in UK and 1 533 239 in USA. In UK population 78 201 (16%) were COX-2 users and in PharMetrics 324 206 (21%) were COX-2 users. Diclofenac and ibuprofen (NSAIDS), and celecoxib and rofecoxib (COX-2) were the agents prescribed most frequently. The duration of therapy was longer among celecoxib and rofecoxib users than among other users. More COX-2 users than NSAIDS users received concomitant gastroprotective agents (GPA), corticosteroids and anti-platelet therapy, and had a history of thromboembolic events and hypertension. PharMetrics patients were prescribed higher doses of NSAIDS and COX-2. The use of any single agent for more than 90 days was uncommon, but more frequent in PharMetrics. Switching was uncommon and was generally to a NSAID. DISCUSSION: Our results confirm some previous findings from other authors such as the presence of both GI and CV channelling to COX-2 agents but refute others, such as the frequency of drug switching between these agents. The typical use of COX-2 agents in practice is for shorter duration, and at lower doses, than was employed in randomized clinical trials. This difference may help clarify the apparent discrepancy with respect to CV toxicity between the results from clinical trials, which showed a higher CV risk with these drugs, and non-experimental epidemiologic studies, which showed lower or no increase in risk. PMID- 17086564 TI - Characterization of peptide-pyrazole interactions in solution by low-temperature NMR studies. AB - Complexation of the amino- and carboxyl-protected tripeptide Piv-L-Val-L-Val-L Val-tBu with 3-methylpyrazole and 3-amino-5-methylpyrazole was studied by low temperature NMR experiments in a freonic solvent. The peptide forms an extended beta-type structure at all temperatures and associates through hydrogen bonding with the two pyrazole-based beta-sheet ligands. A detailed structural characterization of the formed complexes by one- and two-dimensional NMR experiments under slow exchange conditions was made possible by employing very low temperatures. The tripeptide associates to stable antiparallel dimers that are symmetrically capped on both sides by two pyrazole receptors to form 2:2 complexes. Amide groups of two neighboring residues in an extended conformation are involved in cyclic hydrogen bonds to the pyrazole. Based on amide chemical shift changes, the relative strength of intermolecular hydrogen bonds can be assessed and correlated with the electronic effects of the substituents on the pyrazole. PMID- 17086565 TI - Reactions of Sn(NMe2)2 with MPHCy: the effects of alkali metal phosphide coupling (Cy=cyclohexyl; M=Li, Na, K, Rb). AB - The reactions of the SnII base Sn(NMe2)2 with CyPHM (Cy=cyclohexyl) produce a range of products, depending primarily on the alkali metal (M) involved. The 1:3 stoichiometric reaction of Sn(NMe2)2 with CyPHNa in the presence of the Lewis base donor PMDETA (PMDETA=(Me2NCH2CH2)2NMe) gives [(NaPMDETA)2{Sn(mu-PCy)}3] (3), containing the electron-deficient [{Sn(mu-PCy)}3]2- dianion. Natural bond order (NBO) and electron localisation function (ELF) calculations show that this species is described most appropriately by a two-electron, three-centre Sn3 bonding model. Evidence that 3 results from phosphide coupling is provided by the 1:1 reaction of Sn(NMe2)2 with CyPHNa in the presence of PMDETA, which gives 3 and trace amounts of (NaPMDETA)2[{Sn(mu-PCy)}2(mu-PCyPCy)] (4) (containing one PCyPCy2- dianion). Greater extents of phosphide coupling are observed as the size of the Group 1 metal is increased. Thus, the 1:3 reaction of Sn(NMe2)2 with CyPHK in THF gives the co-crystalline product {(K2 THF)2[{Sn(mu-PCyPCy)}2(mu PCy)]}0.9{(K2 THF)2[{Sn(mu-PCy)}2(mu-PCyPCy)]}0.1 (5) (containing [{Sn(mu PCyPCy)}2(mu-PCy)]2- and [{Sn(mu-PCy)}2(mu-PCyPCy)]2- dianions), whereas the analogous reaction of Sn(NMe2)2 with RbPHCy gives [RbPMDETA{(CyP)3SnP(H)Cy}] (6) (containing a cyclic {(CyP)3Sn} unit). PMID- 17086566 TI - Magnetic face-to-face interaction and electrocommunication in chromium sandwich compounds. AB - The reaction of [{(C5Me5)CrCl2}2] with [2.2](1,4)cyclophane gave [(C5Me5)Cr{[2.2](1,4)cyclophane}] (1) and [(C5Me5)Cr{[2.2](1,4)cyclophane}Cr(C5Me5)] (2), depending on the reaction conditions. X-ray structure analysis showed 2 to be a ministack which in turn is stacked in the lattice. The chromium atoms are 6.035 A apart, and the distortion of the benzene rings to boat-shaped moieties is less pronounced than in parent [2.2](1,4)cyclophane. The NMR and EPR spectra were consistent with a S=1/2 ground state for 1 and with two interacting S=1/2 centers in 2. Spin density was found in the ligand pi systems, where its sign was negative when the pi system was adjacent to chromium, while on the nonbonded benzene moiety of 1 it was positive. Cyclic voltammograms showed reductions to 1- and 2(2-), as well as oxidations to 1+, 2+, and 2(2+) which were quasireversible, whereas oxidations to 1(2+) and 2(3+) were irreversible. Interaction between the metal ions was revealed by a 260 mV separation of the redox waves belonging to 2+, and 2(2+). Both cations were isolated as [B(C6H5)4]- salts, which in solution decomposed to [2.2](1,4)cyclophane and [(C5Me5)Cr{(eta6-C6H5)B(C6H5)3}] (3). The 1H and 13C NMR spectra of 3 were in accordance with an S=1 ground state. Solid-state magnetic measurements of the dimetallic compounds showed antiferromagnetic interaction with J=-122 cm-1 for 2, J=-31 cm-1 for 2+ (ground state S=1/2), and J=-23.5 cm-1 for 2(2+) (with H=-JS1S2). The decrease of J in the series 2, 2+, and 2(2+) was traced to the number of unpaired electrons and, for the mixed-valent cation 2+, to additional double exchange. PMID- 17086567 TI - Association between nonnaproxen NSAIDs, COX-2 inhibitors and hospitalization for acute myocardial infarction among the elderly: a retrospective cohort study. AB - PURPOSE: To evaluate the association between rofecoxib, celecoxib, diclofenac, and ibuprofen and the risk of hospitalization for acute myocardial infarction (AMI) in an elderly population. METHODS: We conducted a retrospective cohort study, using data from the government of Quebec health insurance agency databases, among patients 65-80 years of age who filled a prescription for any of the study drugs during 1999-2002. Cox regression models with time-dependent exposure were used to compare the incidence rates of hospitalization for AMI adjusting for patients' baseline characteristics. Analyses stratified by dose and number of supplied days were also conducted. RESULTS: At the index date, a total of 91 062 patients were taking rofecoxib, 127 928 celecoxib, 49 193 diclofenac, and 15 601 ibuprofen. The adjusted hazard ratio (HR) (95%CI) of hospitalization for AMI were: celecoxib versus rofecoxib: 0.90 (0.79, 1.01); ibuprofen versus rofecoxib: 0.95 (0.65, 1.37); diclofenac versus rofecoxib: 1.01 (0.84, 1.22). In secondary analyses based on intended duration of use, neither COX-2 selective inhibitor was associated with a higher risk than ibuprofen or diclofenac. The unadjusted risk of AMI for all NSAIDs increased with dose. In the direct two way adjusted comparison of each NSAID stratified by dose, the only statistically significant difference was with rofecoxib >25 mg/day versus celecoxib >200 mg/day. CONCLUSION: In this study there was no difference between AMI occurrence in elderly patients taking rofecoxib or celecoxib at recommended doses for chronic indications versus those taking ibuprofen/diclofenac. However, the risk of AMI was higher among patients using higher doses of rofecoxib (>25 mg/day) compared to patients using higher doses of celecoxib (>200 mg/day). PMID- 17086568 TI - X-Linked dominant chondrodysplasia punctata: prenatal diagnosis and autopsy findings. AB - OBJECTIVE: To report our experience of the prenatal diagnosis of X-linked dominant chondrodysplasia punctata (CDPX2) and highlight its variable phenotypic presentation. METHODS: We report the sonographic features of three female fetuses affected with CDPX2. The ultrasound, radiographic and pathological findings were compared. RESULTS: Family 1: Two affected pregnancies, both terminated. Fetus 1: Presented with epiphyseal stippling involving the vertebrae, upper and lower limbs, asymmetric shortening of the long bones and flat facial profile. Fetus 2: Prenatal findings included premature epiphyseal stippling, paravertebral cartilaginous calcific foci, mild shortening of the long bones and flat facies. Mutation analysis of the mother and both fetuses revealed mutation in the emopamil-binding protein (EBP) gene. Family 2: Prenatal sonography showed scattered epiphyseal stippling, minimal vertebral segmentation anomalies, mild asymmetric limb shortening and flat facies. Female infant delivered at 39 weeks of gestation. Biochemical analysis in all three fetuses showed increased levels of serum 8(9)-cholestenol consistent with delta (8), delta (7)-isomerase deficiency and CDPX2. CONCLUSION: Prenatal diagnosis of CDPX2 is difficult because of marked phenotypic variation. Epiphyseal stippling, ectopic paravertebral calcifications, asymmetric shortening of long bones and dysmorphic flattened facies are crucial for prenatal diagnosis. DNA analysis of the CDPX2 gene and biochemical determination of the serum 8(9)-cholestenol level are important for diagnosis, especially if future pregnancies are planned. PMID- 17086569 TI - Exploring the dynamics of calix[4]pyrrole: effect of solvent and fluorine substitution. AB - Molecular dynamics simulations show that calix[4]pyrrole (CP) and octafluorocalix[4]pyrrole (8F-CP) are extremely flexible molecules. CP mainly adopts the 1,3-alternate conformation in all the solvents, although the percentage of alternative conformations increases in polar solvents, especially those with good hydrogen-bonding acceptor properties. However, in the case of 8F CP, the cone conformation is the most populated in some solvents. Transitions between conformers are common and fast, and both CP and 8F-CP can adopt the cone conformation needed for optimum interaction with anions more easily than would be predicted on the basis of previous gas-phase calculations. Furthermore, the present studies show that when a fluoride anion is specifically placed initially in close proximity to CP and 8F-CP in their respective 1,3-alternate conformations, an extremely fast change to the cone conformation is observed in both cases. The results suggest that preorganization does not represent a major impediment to anion-binding for either CP or 8F-CP, and that ion-induced conformational changes can follow different mechanisms depending on the solvent and the chemical substituents present on the calix[4]pyrrole beta-pyrrolic positions. PMID- 17086570 TI - Design, synthesis, and delivery properties of novel guanidine-containing molecular transporters built on dimeric inositol scaffolds. AB - We have developed a novel class of synthetic molecular transporters that contain eight residues of guanidine with an inositol dimer as the scaffold. The dimers were prepared by connecting two units of myo- or scyllo-inositol via a carbonate or amide linkage, and the multiple units of the guanidine functionality were constructed on the inositol scaffold by means of peracylation with omega aminocarboxylate derivatives of varying length. Bioassays based on confocal laser scanning microscopy and fluorescence-activated cell sorter analyses indicated that these transporters display a varying degree of membrane translocating ability, and the intracellular localization and mouse-tissue distribution studies strongly suggested that these transporters undergo substantially different mechanistic processes from those of peptide transporters reported to date. It was also shown that doxorubicin, an anticancer antibiotic, can be efficiently delivered into mouse brain by aid of this type of transporter. PMID- 17086571 TI - Asymmetric synthesis of rubiginones A(2) and C(2) and their 11-methoxy regioisomers. AB - Convergent enantioselective syntheses of angucyclinone-type natural products rubiginones A(2) (2) and C(2) (1) and their 11-methoxy regioisomers 3 a and 3 b have been achieved by using two domino processes from a common enantiomerically pure 1-vinylcyclohexene 4. Key steps in the synthesis of this diene were the stereoselective conjugate addition of AlMe(3) on (SS)-[(p-tolylsulfinyl)methyl]-p quinol (9) and the elimination of the beta-hydroxy sulfoxide fragment, after oxidation to sulfone, to recover a carbonyl group. The first domino sequence comprised Diels-Alder reaction with a sulfinyl naphthoquinone followed by sulfoxide elimination. An efficient opposite regioselection in the cycloaddition step was achieved in the convergent construction of the tetracyclic skeleton using a sulfoxide at C-2 or C-3 of the dienophiles 5 or 6, derived from 5-methoxy 1,4-naphthoquinone. The second domino process, triggered by oxygen and sunlight, allowed the transformation of the initial tetracyclic adducts into the final products after B ring aromatization, silyl deprotection and C-1 oxidation. PMID- 17086572 TI - An analysis of the psychometric profile and frequency of anxiety and depression in Australian men with prostate cancer. AB - Zung's Self-Rating Anxiety Scale and Self-Rating Depression Scale were used to collect data from 195 Australian men who had received a diagnosis of prostate cancer. Analysis was via separate scales and also by combining both scales into a single unit to assess anxiety-depression and then analysing the underlying component structure of that unit. Applying Zung's recommended cutoff scores, 12% of the sample were classified as having clinically significant levels of anxiety and 16% had similar levels of depression. Factor analysis of the combined SAS and SDS indicated four major components which reflected a process of: loss of functional capacity, worthlessness and hopelessness, fear and somatic symptomatology. Implications for the effective assessment and treatment of anxiety and depression in prostate patients are discussed. PMID- 17086573 TI - Structural, solid-state NMR and theoretical studies of the inverse-coordination of lithium chloride using group 13 phosphide hosts. AB - The reaction of MeAlCl2 with 'PhPLi2' in THF gives [{MeAl(PPh)3Li(4).3 THF}4(mu4 Cl)]-Li+ (1). The GaIII and InIII analogues, [{MeE(PPh)3Li(4).3 THF}4(mu4-Cl)] Li+(THF)3 (E=Ga (2), In (3)), are obtained by the in situ reactions of MeECl2 with PhPLi2 in THF. For all of the complexes, the cage anions have an unusual cubic arrangement that is similar to a zeolite, and contain large voids (ca. 17 A). The location of the Li+ counterions in 1-3 and their coordination environment appears to subtly reflect variations in packing and lattice energy. Whereas in 1 highly mobile, loosely coordinated Li+ counterions are present, 2 and 3 contain less mobile THF-solvated counterions within the cavities. X-ray crystallographic and solid-state NMR studies are reported on 1-3, together with model DFT calculations on the selectivity of halide coordination. PMID- 17086574 TI - The effect of counterion/ligand interplay on the activity and stereoselectivity of palladium(II)-diimine catalysts for CO/p-methylstyrene copolymerization. AB - The catalytic activity and stereoselectivity of complexes [Pd(eta(1),eta(2) C(8)H(12)OMe)(Ar--N==C(R')--C(R')==N--Ar)]X in the copolymerization of CO and p methylstyrene have been correlated with their interionic structure in solution and in the solid state, as determined by (19)F,(1)H-HOESY NMR spectroscopy and X ray diffraction studies, respectively. The highest productivity is obtained with unhindered diimine ligands bearing electron-donating substituents and with the least coordinating counterion. Copolymers with a microstructure ranging from atactic to predominantly isotactic are obtained. The degree of isotacticity increases as the steric hindrance in the apical positions and the coordinating ability of the counterion increase. The counterion is located close to the diimine in both solution and the solid state but it moves toward the palladium as the steric hindrance in the apical positions decreases. When the latter is small the counterion competes with the substrate for apical coordination, and consequently it affects the productivity. In the case of ortho-dimethyl substituted ligands the counterion is confined in the back, above the N==C(R')- C(R')==N moiety, and does not affect the productivity. However, it contributes to increasing the stereoregularity of the copolymer by making the aryl moieties more rigid. With R'=Me and Ar=o-Me(2)C(6)H(3) an ll of 81 % and 72 % was obtained with X(-)=CF(3)SO(3) (-) or BArF(-), respectively. The isotacticity of the copolymers produced by ortho-monosubstituted catalysts depends greatly on the counterion and ranges from 30 % to 59 % with X(-)=BArF(-) and X(-)=CF(3)SO(3) (-), respectively, with Ar=o-EtC(6)H(4) and R'=Me. Based on the interionic structural results, this effect can be explained by a greater reduction of the copolymerization rate of C(s)-symmetric isomers with respect to their C(2)-symmetric counterparts. PMID- 17086575 TI - Glycosidation-anomerisation reactions of 6,1-anhydroglucopyranuronic acid and anomerisation of beta-D-glucopyranosiduronic acids promoted by SnCl(4). AB - The reaction of silylated nucleophiles with 6,1-anhydroglucopyranuronic acid (glucuronic acid 6,1-lactones) catalysed by tin(IV) chloride provides 1,2-trans or 1,2-cis (deoxy)glycosides in a manner dependent on the donor structure. The alpha-glycoside was obtained for reactions of the donor with the 2-acyl group and 2-deoxydonors, whereas the 2-deoxy-2-iodo donor gave the beta-glycoside. Experimental evidence shows that when 1,2-cis-glycoside formation occurs, the anomerisation of initially formed 1,2-trans-glycosides catalysed by SnCl(4) is possible. The anomerisation of beta-D-glucopyranosiduronic acids was found to be faster, in some cases, than anomerisation of related beta-D-glucopyranosiduronic acid esters and beta-D-glucopyranoside derivatives and the rates are dependent on the structure of the aglycon. Moreover, the rates of anomerisation of beta-D glucopyranuronic acid derivatives can be qualitatively correlated with rates of hydrolysis of beta-D-glucopyranosiduronic acids. Mechanistic possibilities for the reactions are considered. PMID- 17086576 TI - Imaging with chemical analysis: adsorbed structures formed during surface chemical reactions. AB - Imaging surfaces and interfaces with structural and chemical specificity has been essential for understanding a variety of phenomena occurring in adsorbed layers during surface chemical reactions. A recent achievement of chemical imaging with spectroscopic analysis is the experimental proof of theoretically predicted spontaneous formation of regular patterns of metal adatoms during surface chemical reactions. An attractive feature of this finding is that the reaction rate and adlayer coverage can be employed to precisely control the morphology of the structures. The mechanisms of these self-organisation phenomena, driven by the interplay between energetic principles and kinetics, opens a conceptually novel route to creating a wide range of surface-supported functional structures at the micro- and nanometre length scales. PMID- 17086578 TI - Right aortic arch detected in fetal life. AB - OBJECTIVE: To evaluate the prenatal distribution, associated conditions and outcome of the different types of right aortic arch (RAA) detected in fetal life. METHODS: This was a retrospective review of all cases of RAA detected prenatally between 1998 and 2005 in two tertiary referral centers. RESULTS: In the study period 71 cases of RAA were detected; 26 (37%) had RAA with aberrant left subclavian artery, 23 (32%) had RAA with mirror-image branching, 20 (28%) had RAA of unknown type and two (3%) had double aortic arch. While 20/26 cases with RAA and aberrant left subclavian artery were isolated findings, all 23 cases with RAA and mirror-image branching were associated with cardiac defects, namely tetralogy of Fallot (43%) or pulmonary atresia with ventricular septal defect (22%). Of the 20 cases with RAA, 19 of unknown type were associated with heterotaxy syndromes and had additional cardiac malformations and ambiguities of the situs. The two cases with DAA were isolated findings. Seven cases in our series (10%) had a microdeletion 22q11 and these were significantly associated with extracardiac malformations. The outcome in our series depended solely on the associated cardiac and extracardiac malformations, with the exception of one infant with isolated DAA, in whom a surgical correction was warranted. CONCLUSIONS: RAA detected in fetal life is associated frequently with other cardiac/non-cardiac malformations, heterotaxy syndromes and microdeletions 22q11. The associated conditions vary depending on the branching type of the brachiocephalic vessels and the presence of extracardiac malformations. PMID- 17086579 TI - Brachycephaly and frontal lobe hypoplasia in fetuses with trisomy 21 at 11 + 0 to 13 + 6 weeks. AB - OBJECTIVE: To investigate the incidence of brachycephaly and frontal lobe hypoplasia in fetuses with trisomy 21 at 11 + 0 to 13 + 6 weeks of gestation. METHODS: A three-dimensional (3D) volume of the fetal head was obtained before fetal karyotyping at 11 + 0 to 13 + 6 (median, 12 + 5) weeks of gestation in 100 fetuses that were subsequently found to have trisomy 21 and in 300 fetuses subsequently found to be chromosomally normal. The multiplanar mode was used to obtain a sequence of transverse views of the fetal head and to demonstrate the biparietal and suboccipitobregmatic views. We measured the biparietal diameter (BPD), the occipitofrontal diameter (OFD) and the frontothalamic distance (FTD) between the inner table of the frontal bone and the posterior thalami. RESULTS: In the chromosomally normal group the BPD, OFD and FTD increased linearly with crown-rump length (CRL) from 16.7 mm, 19.0 mm and 12.1 mm at a CRL of 45 mm to 26.7 mm, 31.7 mm and 18.7 mm, respectively, at a CRL of 84 mm. In the trisomy 21 fetuses, compared to normal fetuses, there was shorter BPD (mean difference = 0.63 mm; 95% CI, -0.97 to -0.30 mm, P < 0.0001), OFD (mean difference = -1.41 mm; 95% CI, -1.75 to -1.07 mm, P < 0.0001) and FTD (mean difference = -0.77 mm; 95% CI, -1.02 to -0.54 mm; P < 0.0001) and higher BPD to OFD ratio (mean difference = 0.022; 95% CI, 0.012 to 0.032, P < 0.0001) but no significant difference in the FTD to OFD ratio (mean difference = 0.004; 95% CI, -0.006 to 0.013, P = 0.448). CONCLUSIONS: In fetuses with trisomy 21 at 11 + 0 to 13 + 6 weeks of gestation there is evidence of brachycephaIy but not of frontal lobe hypoplasia. PMID- 17086580 TI - Associated and predictive factors of sleep disturbance in advanced cancer patients. AB - Little attention has been paid to sleep disturbance experienced by advanced cancer patients. The purpose of the present study was to investigate longitudinal change in sleep disturbance and to identify factors that associated with and predicted sleep disturbance among 209 consecutive terminally ill cancer patients. Patients were assessed twice for sleep disturbance by one item of the structured clinical interview for assessing depression, once at the time of their registration with a palliative care unit (PCU) (baseline) and again at the time of their PCU admission (follow-up), and possible associated medical and psychosocial factors were evaluated. The proportions of patients with obvious sleep disturbance at baseline and follow-up were 15.3 and 25.9%, respectively. Sixty-seven percent of the subjects showed some sleep status changes, including both aggravation and improvement, between baseline and follow-up. Being younger, having diarrhea and living alone were significantly associated with sleep disturbance at baseline, and the increase of psychological distress was the only significant predictive factor for sleep disturbance at follow-up. These findings suggest that psychological distress is a possible key cause of sleep disturbance and management of psychological distress may be one promising strategy for prevention of sleep disturbance among advanced cancer patients. PMID- 17086581 TI - Prenatal diagnosis of open and closed spina bifida. AB - OBJECTIVE: To identify criteria useful for differentiating closed from open spina bifida antenatally. PATIENTS AND METHODS: A retrospective study of cases of spina bifida diagnosed in a referral center between 1997 and 2004. RESULTS: Of 66 cases of fetal spina bifida diagnosed at a median gestational age of 21 (range, 16-34) weeks, detailed follow-up was available for 57. Of these, open defects were found in 53 (93.0%) and closed defects in four (7.0%). Closed spina bifida was associated in two cases with a posterior cystic mass with thick walls and a complex appearance, while in two cases the spinal lesion could not be clearly differentiated from an open defect, particularly at mid-gestation. Open spina bifida was always associated with typical alterations of cranial anatomy, including the so-called 'banana' and 'lemon' signs, while in closed spina bifida the cranium was unremarkable. When the data were available, levels of amniotic fluid alpha-fetoprotein were always abnormally elevated with open spina bifida and within normal limits with closed forms. CONCLUSION: In this study 7% of cases of spina bifida diagnosed in utero were closed. The differentiation between open and closed forms is best shown by the sonographic demonstration of abnormal or normal cranial anatomy. PMID- 17086582 TI - Fetal pleuroamniotic shunting for bronchopulmonary sequestration with hydrops. AB - Bronchopulmonary sequestration (BPS), a non-functional pulmonary tissue mass, when complicated by fetal hydrops, carries a high risk of perinatal mortality. However, a limited number of cases of BPS complicated by fetal hydrops with an informative clinical course have been reported. We report here on three cases of BPS complicated by fetal hydrothorax and hydrops that were successfully treated by pleuroamniotic shunting, which should be considered as a treatment option for fetal hydrothorax and hydrops associated with BPS. PMID- 17086583 TI - Predicting pulmonary hypoplasia using the sonographic fetal lung volume to body weight ratio--how precise and accurate is it? AB - OBJECTIVES: To determine the precision and accuracy of ultrasound in estimating the fetal lung to body weight ratio (FLB ratio) using two-dimensional (2D) and three-dimensional (3D) ultrasound by comparison with postmortem measurements, and to evaluate its potential to diagnose pulmonary hypoplasia. METHODS: Lung volumes were estimated by 3D ultrasound (rotational technique) and fetal weights were measured by 2D ultrasound (Hadlock equation) in 35 fetuses immediately before termination of pregnancy at 15-38 weeks. Sonographic estimates of FLB ratio were compared with postmortem values. Based on the pathological definition of pulmonary hypoplasia, the accuracy of sonographic estimation of the FLB ratio was analyzed. RESULTS: The mean gestational age at termination of pregnancy was 26.7 (range, 15-38) weeks. The mean FLB ratios were 0.018 (SD, 0.006) on ultrasound and 0.019 (SD, 0.007) at autopsy (P = 0.730). Bias and precision of sonographic FLB ratio were - 0.001 and 0.003 (absolute limits, - 0.007 to + 0.006), respectively. Pulmonary hypoplasia was diagnosed in 12 (34.3%) cases at autopsy. The sonographic FLB ratio was significantly lower in fetuses with pulmonary hypoplasia at autopsy (median, 0.011; range, 0.004-0.014) than it was in those without pulmonary hypoplasia (median, 0.022; range, 0.013-0.045, P < 0.001). The sensitivity and specificity of the sonographic FLB ratio for diagnosing pulmonary hypoplasia were 91.7% (11/12) and 91.3% (21/23), respectively, the positive and negative predictive values were 84.6% (11/13) and 95.5% (21/22), and the accuracy was 91.4% (32/35). CONCLUSION: FLB ratio can be estimated precisely on ultrasound examination, albeit with wide limits of agreement. The sonographically estimated FLB ratio may be useful in the prediction and diagnosis of pulmonary hypoplasia. PMID- 17086585 TI - Self-assembly of nanometer-scale [Cu24I10L12]14+ cages and ball-shaped Keggin clusters into a (4,12)-connected 3D framework with photoluminescent and electrochemical properties. PMID- 17086584 TI - Reactions in droplets in microfluidic channels. AB - Fundamental and applied research in chemistry and biology benefits from opportunities provided by droplet-based microfluidic systems. These systems enable the miniaturization of reactions by compartmentalizing reactions in droplets of femoliter to microliter volumes. Compartmentalization in droplets provides rapid mixing of reagents, control of the timing of reactions on timescales from milliseconds to months, control of interfacial properties, and the ability to synthesize and transport solid reagents and products. Droplet based microfluidics can help to enhance and accelerate chemical and biochemical screening, protein crystallization, enzymatic kinetics, and assays. Moreover, the control provided by droplets in microfluidic devices can lead to new scientific methods and insights. PMID- 17086586 TI - Single-molecule protein encapsulation in a rigid DNA cage. PMID- 17086587 TI - Methane oxidation by aqueous osmium tetroxide and sodium periodate: inhibition of methanol oxidation by methane. PMID- 17086589 TI - Defining and measuring optical frequencies: the optical clock opportunity--and more (Nobel lecture). AB - Four long-running currents in laser technology met and merged in 1999-2000. Two of these were the quest toward a stable repetitive sequence of ever-shorter optical pulses and, on the other hand, the quest for the most time-stable, unvarying optical frequency possible. The marriage of ultrafast- and ultrastable lasers was brokered mainly by two international teams and became exciting when a special "designer" microstructure optical fiber was shown to be nonlinear enough to produce "white light" from the femtosecond laser pulses, such that the output spectrum embraced a full optical octave. Then, for the first time, one could realize an optical frequency interval equal to the comb's lowest frequency, and count out this interval as a multiple of the repetition rate of the femtosecond pulse laser. This "gear-box" connection between the radiofrequency standard and any/all optical frequency standards came just as sensitivity-enhancing ideas were maturing. The four-way union empowered an explosion of accurate frequency measurement results in the standards field and prepared the way for refined tests of some of our cherished physical principles, such as the time-stability of some of the basic numbers in physics (e.g. the "fine-structure" constant, the speed of light, certain atomic mass ratios), and the equivalence of time-keeping by clocks based on different physics. The stable laser technology also allows time synchronization between two independent femtosecond lasers so exact they can be made to appear as if the source were a single laser. By improving pump-probe experiments, one important application will be in bond-specific spatial scanning of biological samples. This next decade in optical physics should be a blast! PMID- 17086590 TI - A molecular dynamics study of glucose solvation in the ionic liquid 1,3 dimethylimidazolium chloride. PMID- 17086591 TI - Coinage metal halide clusters: from two-dimensional ring to three-dimensional solid-state-like structures. PMID- 17086592 TI - Formation of trichlorinated dibenzo-p-dioxins from 2,4-dichlorophenol and 2,4,5 trichlorophenolate: a theoretical study. AB - The reaction of the 2,4,5-trichlorophenolate anion with 2,4-dichlorophenol to afford trichlorinated dibenzo-p-dioxins (T3CDDs) is investigated at the B3LYP/6 31+G(d) and B3LYP/6-311+G(3df,2p)//B3LYP/6-31+G(d)+ZPVE(B3LYP/6-31+G(d)) levels of theory. The first stage of the process corresponds to the formation of a predioxin, which can evolve through four different routes. Two of them lead directly to the products 2,3,7-T3CDD and 1,3,8-T3CDD, and the other two afford different predioxin-type intermediates, which in turn can evolve through all or some of the four routes to give new predioxins or T3CDD. Consequently, the theoretical results obtained show plainly the complex chemistry implied in the formation of dioxins from chlorophenols via anionic mechanisms by disclosing all the critical structures through which the system evolves, thus allowing assessment of the viability of the different mechanistic routes and the accessible products. The statistical thermodynamics treatment at 1 atm and 298.15, 600, 900, and 1200 K indicates that at higher temperatures, the Gibbs energy barrier for the formation of the initial predioxin is clearly the rate determining step for the whole process, but at lower temperatures the Gibbs energy barrier for this step is similar to those for its evolution into 2,3,7 T3CDD. This result is in contrast with previous proposals that the closure of the central ring is the rate-limiting step. Finally, according to our results the rate constant for the formation of polychlorinated dibenzo-p-dioxins increases with the temperature, in agreement with the experimental observation that the conversion of trichlorophenols increases when going from 600 to 900 K in the gas phase in the absence of catalysts, and with DFT molecular dynamics results. PMID- 17086593 TI - Direct experimental observation of CS2OH. AB - The first experimental detection of CS(2)OH is reported. CS(2)OH was observed for about one microsecond after its formation, as an intact isolated species in the gas phase. It was generated by electron transfer to the CS(2)OH(+) ion, prepared in the source of a multisector mass spectrometer by suitable ion-molecule reactions. The vertical formation process allowed characterization of CS(2)OH by structural analysis of CS(2)OH(+). Theoretical calculations were performed at the B3LYP/6-311+G(2d,p) and CCSD(T)/aug-cc-pVTZ//B3LYP/6-311+G(2d,p) levels of theory. The computed structure and stability of CS(2)OH and CS(2)OH(+) as well as the energetics of the involved processes satisfactorily fit with the experimental results. PMID- 17086594 TI - Theoretical determination of the vibrational Raman optical activity signatures of helical polypropylene chains. AB - Raman and vibrational Raman optical activity (VROA) spectra of helical conformers of polypropylene chains are simulated using ab initio methods to unravel the relationships between the vibrational signatures and the primary and secondary structures of the chains. For a polypropylene chain containing three units, conformational effects are shown to lead to more acute signatures for VROA than for Raman spectra. In addition to regular polypropylene chains, which can display right and left helicities with the same probability, chirality and therefore helicity are enforced by substituting one chain end with a phenyl group. The simulations predict that the threefold helical structures, which correspond to (TG)(N) conformations of the backbone, have a specific VROA backward signature in the form of an intense couplet around 1100 cm(-1). This couplet is associated with collective wagging and twisting motions, while most of its intensity comes from the anisotropic invariants combining normal coordinate derivatives of the electric dipole-electric dipole polarizability and of the electric dipole magnetic dipole polarizability. A similar signature has already been found in model helical polyethylene chains, whereas it is very weak in forward VROA. PMID- 17086596 TI - Citrullinated antigens: just diagnostic tools or pathogenic targets in rheumatoid arthritis? PMID- 17086597 TI - Cardiovascular risk in inflammatory rheumatic diseases: loose ends and common threads. PMID- 17086598 TI - Improving management of pregnancy in antiphospholipid antibody-positive women. PMID- 17086599 TI - Remember the Titanic: what we know of knee osteoarthritis is but the tip of the iceberg. PMID- 17086600 TI - The Symptom Intensity Scale, fibromyalgia, and the meaning of fibromyalgia-like symptoms. A review. PMID- 17086601 TI - HLA-DRB1*04 alleles in Japanese rheumatoid arthritis patients with AA amyloidosis. AB - OBJECTIVE: To compare the HLA-DRB1 shared epitope (SE) alleles in Japanese patients with rheumatoid arthritis (RA) and amyloid A (AA) amyloidosis versus those without AA amyloidosis. METHODS: The HLA-DRB1 alleles were genotyped for 91 RA patients without AA amyloidosis, 33 RA patients with AA amyloidosis, and 63 control subjects. HLA-DRB1 typing was performed by polymerase chain reaction, sequence-specific oligonucleotide probe hybridization method. RESULTS: Although a significant difference was not observed, the frequency of SE genotype was higher in RA patients with AA amyloidosis than in those without AA amyloidosis. All SE positive RA patients with AA amyloidosis had *04 alleles (*0401, *0405, *0410), and a significant association of the presence of a double dose of *04 SE alleles with AA amyloidosis (OR 4.0, 95% CI 1.91-13.99) was observed. CONCLUSION: Our data suggest that presence of double *04 SE is associated with a higher risk of developing AA amyloidosis in Japanese patients with RA. PMID- 17086602 TI - Cost-effectiveness of biologic agents for treatment of autoimmune disorders: structured review of the literature. AB - OBJECTIVE: Four new biologic treatments have been approved for several autoimmune disorders. Economic evaluations have been used to model their cost-effectiveness. METHODS: We conducted a structured literature review in Embase and PubMed to identify all relevant cost-effectiveness models investigating one or more of these 4 drugs in autoimmune disorders. RESULTS: Fifteen full economic evaluations were identified [13 for rheumatoid arthritis (RA), 2 for Crohn's disease (CD), and 1 for ankylosing spondylitis (AS)]. While several studies found adalimumab, etanercept, and infliximab to be cost-effective (using a threshold around $50,000/quality-adjusted life-year) for treatment of severe RA, not all studies concurred, and there was significant variation in the range of cost-effectiveness ratios reported. Neither study in CD found treatment with infliximab to be cost effective. Only one study was identified in AS: treatment with infliximab was found to be cost-effective. CONCLUSION: Modeling treatment strategies in chronic relapsing diseases such as RA, CD, and AS presents particular challenges, as reflected in the variation in cost-effectiveness results reported. A reference case for economic evaluations, such as that suggested by the OMERACT (Outcome Measures in Rheumatology) Health Economics Working Group will facilitate comparison and interpretation of results. PMID- 17086603 TI - Association of rheumatoid arthritis with ergothioneine levels in red blood cells: a case control study. AB - OBJECTIVE: The dietary thiol compound and erythrocyte ingredient ergothioneine (ET) is the preferential physiological substrate of the organic cation transporter OCTN1, found to be associated with rheumatoid arthritis (RA) in genetic studies, but the biological roles of ET and OCTN1 are unclear. We investigated the association between ET concentrations in peripheral blood erythrocytes and the occurrence of RA. METHODS: Erythrocyte ET concentrations in patients with mildly active RA (n = 73) were compared to ET levels in patients with coronary heart disease (CHD; n = 62) and osteoarthritis (OA; n = 148), serving as non-RA chronic inflammatory disease controls. Correlation of ET levels in erythrocytes with levels of ET and OCTN1 mRNA in CD14+ monocytes was determined in 10 healthy subjects. RESULTS: Erythrocyte ET levels were significantly higher in patients with RA, with a median (interquartile range) of 12.6 micromole/l of erythrocytes (IQR 8.1-18.3), compared to 7.7 (IQR 5.0-12.0; p < 0.001) in CHD and 7.8 (IQR 4.8-12.8; p < 0.001) in OA. The prevalence of RA compared to non-RA controls increased with increasing blood ET concentrations, with an odds ratio of 0.23 (95% CI 0.13-0.41; p < 0.001) in the lowest quartile of RA erythrocyte ET levels to 3.11 (95% CI 1.54-6.29; p = 0.002) in the highest quartile. The group differences in ET values were maintained after adjustment for disease-related anthropometric and clinical variables (age, sex, body mass index, smoking, duration of disease, hemoglobin, C-reactive protein, and medication) and were also independent of erythrocyte glutathione levels and of polymorphisms of the OCTN1 gene. ET levels in erythrocytes were linearly correlated with ET concentrations (R2 = 0.936, p < 0.001) and OCTN1 mRNA levels (R2 = 0.946, p < 0.001) in CD14+ cells. CONCLUSION: Mildly active cases of RA are associated with an unexplained high level of ET in red blood cells. PMID- 17086604 TI - Echocardiographically guided pericardiocentesis for treatment of clinically significant pericardial effusion in rheumatoid arthritis. AB - OBJECTIVE: To assess the safety and efficacy of echocardiographically guided pericardiocentesis for patients with rheumatoid arthritis (RA) and hemodynamically significant pericardial effusion. METHODS: We identified 16 patients with RA who underwent 18 echocardiographically guided pericardiocentesis procedures at our institution over a 20-year period. Clinical and laboratory characteristics of the patients, response to treatment, complications, and need for future pericardial surgery were abstracted from the echocardiography database. RESULTS: Ten patients were men and 6 were women (mean age, 62 yrs; range, 36-75 yrs). On average, patients were diagnosed with RA 11 years before pericardial disease developed. Twelve of 15 patients were seropositive for rheumatoid factor, 10 patients had radiographic evidence of erosions, and 7 patients had rheumatoid nodules. Cardiac tamponade was present in 11 of the 18 cases. Mean volume drained on the first pericardiocentesis was 504 +/- 264 ml (range 120-1000 ml). The fluid was an exudate with a mean protein concentration of 5 g/dl (range 3.3-51.1 g/dl). All cultures and cytologic findings were negative for bacteria and neoplastic cells. No serious complications resulted from echocardiographically guided pericardiocentesis. For 11 patients, a catheter was placed for intermittent drainage over an average of 3 days. Seven patients ultimately required a more definitive surgical procedure. CONCLUSION: Echocardiographically guided pericardiocentesis is a safe and effective treatment for this uncommon but serious complication of RA. PMID- 17086605 TI - Migraine during systemic lupus erythematosus: findings from brain single photon emission computed tomography. AB - OBJECTIVE: Headache in systemic lupus erythematosus (SLE) is controversial, as is evidence of brain impairment in patients with SLE and headache. Perfusion single photon emission computed tomography (SPECT) was performed to investigate brain impairment in SLE patients with migraine-like headache either from the period of diagnosis or later in the course of disease. METHODS: Eighteen patients with SLE (mean age 40.8 +/- 13.6 yrs) matching these characteristics underwent brain SPECT with 99mTc-HMPAO in the interictal period. Electroencephalography (EEG) and magnetic resonance imaging (MRI) were performed in 12 and 10 patients, respectively. SPECT was analyzed through visual and asymmetry combined analysis as well as by voxel-based statistical analysis compared to a control group of matched healthy subjects (statistical threshold: p = 0.01). RESULTS: Focal hypoperfusion was evidenced in 15 (83%) patients, often matching the main side of pain location, whereas EEG and MRI each gave a positive result in 50% of cases. Using voxel-based analysis, significant hypoperfusion was found in 8 (44%) patients, either lateralized to one side or localized to the anterior cingulate cortex, independent of pain location. CONCLUSION: Brain perfusion SPECT is a sensitive tool for identifying brain impairment in SLE-related migraine, although the mechanisms of brain damage remain to be elucidated. Besides confirming focal hypoperfusion in some patients, in 4 patients statistical analysis revealed interictal hypofunction of the anterior cingulate cortex, a key structure for cortical elaboration of pain in the midline network. PMID- 17086607 TI - Autoimmune disease aggregation in families with primary Sjogren's syndrome. AB - OBJECTIVE: Diverse autoimmune diseases may coexist in the same individual and in families, implying a common etiology. We examined the aggregation of autoimmune diseases among first-degree relatives (FDR) of patients with primary Sjogren's syndrome (pSS). METHODS: This was a population-based case-control family study in which 101 families of women classified as having pSS according to the revised American-European criteria and 124 families of matched controls without autoimmune disease were enrolled to investigate the presence of autoimmune diseases. We performed a genetic analysis that included familial correlation and recurrent risk ratios. RESULTS: In family cases, 38% had at least one FDR with an autoimmune disease, versus 22% in control families [odds ratio (OR) 2.2, 95% confidence interval (CI) 1.2-3.9, p = 0.01]. An autoimmune disease was registered for 7.3% of 876 patients' FDR as compared with 3.85% of 857 controls' FDR (OR 1.97, 95% CI 1.28-3.03, p = 0.002). The most frequent autoimmune diseases registered among the pSS patients' FDR were autoimmune thyroid disease (AITD), systemic lupus erythematosus, and rheumatoid arthritis, which disclosed aggregation. The proband phenotype (i.e., pSS) was correlated with AITD, systemic sclerosis, and all autoimmune diseases when considered together as a trait. Maternal transmission of the autoimmunity trait was observed in cases but not in controls. CONCLUSION: Our results indicate that autoimmune diseases cluster within families of patients with pSS. This familial aggregation of autoimmune diseases adds further evidence that clinically different autoimmune phenotypes might share common susceptibility gene variants, which acting in epistatic pleitropy may represent risk factors for autoimmunity. PMID- 17086606 TI - Ibuprofen fails to prevent brain pathology in a model of neuropsychiatric lupus. AB - OBJECTIVE: Neurologic and psychiatric manifestations are severe complications of systemic lupus erythematosus (SLE). As commonly seen in patients, spontaneous development of lupus-like disease in MRL-lpr mice is accompanied by brain atrophy and behavioral dysfunction. We examined inflammatory and ultrastructural aspects of central nervous system (CNS) involvement using a nonselective cyclooxygenase-2 (COX-2) inhibitor and measuring effects on behavior, microglial activation, and neuronal morphology. METHODS: Ibuprofen (IBU) was provided in a rodent chow (375 ppm) for animals 5-19 weeks of age. Exploration of a novel environment and performance in the forced swim test assessed effects on behavior. Immunohistochemistry, fluoro-Jade B (FJB) staining, and flow cytometry were employed in neuropathological analysis. Transmission electron microscopy was used to examine ultrastructural morphology of cortical, hippocampal, hypothalamic, nigral, and cerebellar cells. RESULTS: Chronic IBU treatment failed to normalize immune status, behavior, and brain mass in lupus-prone MRL-lpr mice. It also did not reduce density of CD3+ lymphocytes in the choroid plexus, or FJB+ neurons in the hypothalamus. Activated F4/80+ microglia increased with age, but IBU treatment was not effective in reducing their numbers. Although numerous dark cells were seen in functionally critical brain regions (e.g., paraventricular nucleus and subgranular zone), ultrastructural morphologies of classical apoptosis or necrosis were not detected. CONCLUSION: The COX-dependent pathway does not seem to be critical in the etiology of CNS disease in this model of neuropsychiatric lupus. Reduced brain mass, increased microglial activation, and condensation of cytoplasm point to a metabolic perturbation (e.g., excitotoxic damage) that compromises function and survival of central neurons during lupus like disease. PMID- 17086608 TI - Lower adrenocortical and adrenomedullary responses to hypoglycemia in premenopausal women with systemic sclerosis. AB - OBJECTIVES: To evaluate function of the hypothalamic-pituitary-adrenal (HPA) axis, adrenomedullary hormonal system (AMHS), and sympathetic noradrenergic system (SNS) in premenopausal women with systemic sclerosis (SSc). METHODS: Insulin-induced hypoglycemia (0.1 IU/kg) was performed in 17 longterm, glucocorticoid-naive SSc patients with low disease activity and in 18 healthy women matched for age and body mass index (BMI). Concentrations of glucose, adrenocorticotrophic hormone (ACTH), cortisol, androstenedione (ASD), dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), 17a hydroxyprogesterone (17OHP), epinephrine (EPI), norepinephrine (NE), interleukin 1ss (IL-1ss), IL-6, and tumor necrosis factor-a (TNF-a) were analyzed in plasma. RESULTS: Basal plasma levels of cortisol, ASD, 17OHP, DHEAS, IL-1ss, IL-6, and TNF-a were not significantly different in SSc compared to controls. Patients had higher basal ACTH (6.76 +/- 1.0 pmol/l in SSc vs 4.14 +/- 0.45 pmol/l in controls; p < 0.05), lower basal DHEA (9.02 +/- 1.64 nmol/l in SSc vs 17.0 +/- 2.8 nmol/l in controls; p < 0.05), and lower basal NE (1.61 +/- 0.26 nmol/l in SSc vs 2.57 +/- 0.38 nmol/l in controls; p < 0.05). Patients had comparable responses of glucose and ACTH to hypoglycemia. General linear model for repeated measurements, with BMI and age as covariates, revealed that the responses of 17OHP (p < 0.05), ASD (p < 0.05), DHEA (p < 0.01), EPI (p < 0.001), and NE (p < 0.001) to hypoglycemia were lower in SSc compared to controls. Cortisol response to hypoglycemia tended to be lower in SSc patients (p = 0.06) compared to controls. CONCLUSION: Our data indicate decreased adrenocortical and adrenomedullary functions in premenopausal women with SSc. Whether the observed changes in the neuroendocrine system are secondary to chronic disease deserves further investigation. PMID- 17086609 TI - Is chronic pain in adulthood related to childhood factors? A population-based case-control study of young adults. AB - OBJECTIVE: To investigate whether recalled childhood pain experiences and illnesses are associated with chronic pain in young adults. METHODS: A cross sectional population-based survey recruited participants aged 18-25 years for a case-control study and obtained information on current pain and recalled childhood experiences. In total, 858 respondents were classified as either non pain controls (n = 276), non-chronic pain cases (pain for < or = 3 months in the previous 6 months, n = 435), or chronic pain cases (pain of > 3 months' duration, n = 119). RESULTS: 858 young adults responded to the survey (adjusted response rate 37%). Of the recalled exposures in childhood, family members with pain (OR 2.48, 95% CI 1.48, 4.15), having more than 2 relatives with pain during childhood (OR 3.03, 95% CI 1.44, 6.40), being admitted to hospital during childhood (OR 1.71, 95% CI 1.04, 2.80), and having more illness than one's peer group at secondary school (OR 3.98, 95% CI 1.99, 7.96) were significantly associated with having chronic pain as a young adult, after adjustment for age, sex, and current psychological distress scores. Recall bias was assessed by comparing actual and recalled admission to the neonatal intensive care unit, with no significant differences being found between the participating groups. CONCLUSION: Several associations were observed between pain status as a young adult and selected self reported childhood experiences of illness and pain. The role of recall bias cannot be excluded in this retrospective study, but the results emphasize the importance of family and childhood experiences of pain in potentially influencing future adult pain status. PMID- 17086610 TI - Higher prevalence of fibromyalgia in patients infected with human T cell lymphotropic virus type I. AB - OBJECTIVE: . Inflammatory rheumatic conditions including rheumatoid arthritis and Sjogren's syndrome have been reported in individuals infected with human T cell lymphotropic virus type I (HTLV-I). Other chronic lymphotropic virus infections such as hepatitis C and human immunodeficiency virus are associated with fibromyalgia (FM). There are no reports about the association between HTLV-I infection and FM. We evaluated the association between FM and HTLV-I infection. METHODS: We conducted a case-control study with prevalent cases. Ex-blood donation candidates with HTLV-I infection from a blood bank cohort, and healthy blood donors as a control group, were submitted to rheumatologic evaluation to compare the prevalence of FM. The following covariables were also evaluated: other rheumatic diseases, age, sex, personal income, level of education, and depression. RESULTS: One hundred individuals with HTLV-I infection and 62 non infected blood donors were studied. Thirty-eight (38%) HTLV-I infected individuals and 3 (4.8%) individuals from the control group presented the diagnosis of FM (OR 12.05, 95% CI 3.53-41.17). Other rheumatic diseases were also more prevalent in the infected group (37% vs 12.9%; OR 3.80, 95% CI 1.63-8.86). In multivariate analysis adjusted by the covariables, the association between HTLV-I and FM was statistically significant (OR 9.14, 95% CI 2.42-34.52). CONCLUSION: Our study shows a greater prevalence of FM in HTLV-I infected individuals, suggesting that FM may be associated with this viral infection. PMID- 17086611 TI - Method guidelines for Cochrane Musculoskeletal Group systematic reviews. AB - The Cochrane Musculoskeletal Group (CMSG), one of 50 groups of the not-for-profit international Cochrane Collaboration, prepares, maintains, and disseminates systematic reviews of treatments for musculoskeletal diseases. To enhance the quality and usability of systematic reviews, the CMSG has developed tailored methodological guidelines for authors of CMSG systematic reviews. Recommendations specific to musculoskeletal disorders are provided for various aspects of undertaking a systematic review, including literature searching, inclusion criteria, quality assessment, grading of evidence, data collection, and data analysis. These guidelines will help researchers design, conduct, and report results of systematic reviews of trials in the following fields of musculoskeletal health: gout, osteoarthritis, osteoporosis, pediatric rheumatology, rheumatoid arthritis, soft tissue rheumatism, spondyloarthropathy, systemic lupus erythematosus, systemic sclerosis, and vasculitis. Systematic reviews need to be conducted according to high methodological standards. These recommendations on developing and performing a systematic review will help improve consistency among CMSG reviews. PMID- 17086612 TI - Knowledge transfer to clinicians and consumers by the Cochrane Musculoskeletal Group. AB - The Cochrane Musculoskeletal Group (CMSG) is one of 50 groups of the Cochrane Collaboration that prepares, maintains, and disseminates systematic reviews of treatments for musculoskeletal diseases. Once systematic reviews are completed, the next challenge is presenting the results in useful formats to be integrated into the healthcare decisions of clinicians and consumers. The CMSG recommends 3 methods to aid knowledge translation and exchange between clinicians and patients: produce clinical relevance tables, create graphical displays using face figures, and write consumer summaries and patient decision aids. Accordingly, CMSG has developed specific guidelines to help researchers and authors convert the pooled estimates of metaanalyses in the systematic reviews to user-friendly numbers. First, clinical relevance tables are developed that include absolute and relative benefits or harms and the numbers needed to treat. Next, the numbers from the clinical relevance tables are presented graphically using faces. The faces represent a group of 100 people and are shaded according to how many people out of 100 benefited or were harmed by the interventions. The user-friendly numbers are also included in short summaries and decision aids written for patients. The different levels of detail in the summaries and decision aids provide patients with tools to prepare them to discuss treatment options with their clinicians. Methods to improve the effects and usability of systematic reviews by providing results in more clinically relevant formats are essential. Both clinicians and consumers can use these products to use evidence-based information in individual and shared decision-making. PMID- 17086613 TI - Position Development Conference of the International Society for Clinical Densitometry. Vancouver, BC, July 15-17, 2005. AB - Every 2 years the International Society for Clinical Densitometry (ISCD) organizes a position development conference at which a panel of experts reviews topics of interest in the field of bone densitometry and proposes Official Positions of the ISCD. In July 2005, the most recent conference was held in Vancouver, Canada, during which 4 topics were discussed: (1) Cross-calibration of dual energy x-ray absorptiometry (DXA) systems, (2) Vertebral fracture assessment, (3) World Health Organization (WHO) classification of bone density for populations other than postmenopausal women, and (4) WHO classification for varying skeletal sites. The Official Positions reached at this conference enumerated in this report have been published in The Journal of Clinical Densitometry, and can be found at www.iscd.org. PMID- 17086614 TI - Bone involvement in generalized crystal-storing histiocytosis. AB - The abnormal secretion of monoclonal immunoglobulins observed with monoclonal gammopathies and other clonal B cell dyscrasias can be responsible for a spectrum of deposition disorders. Crystal-storing histiocytosis (CSH) is a rare disease affecting patients with B cell dyscrasias and monoclonal gammopathies, characterized by the accumulation of histiocytes that have phagocytosed an abnormal crystalline immunoglobulin. We describe 2 cases of this rare disorder with multiorgan involvement and prominent bone involvement. Magnetic resonance imaging showed bone marrow infiltration and images of avascular necrosis. Bone specimen analysis gave histological proof of diffuse bone infiltration by the abnormal histiocytes. Bone involvement, which appears to be a specific feature of CSH, links this entity to other storage disorders, such as Gaucher disease. Because the accumulation of abnormal immunoglobulin-loaded histiocytes is clearly pivotal, CSH should be considered not only as an immunoglobulin deposition disease but also as a storage histiocytic disorder. PMID- 17086615 TI - Should physicians manipulate their assessment tools? No, they shouldn't! PMID- 17086617 TI - Tumor necrosis factor-alpha as a potential target in the treatment of systemic lupus erythematosus: a role for the HMG-CoA reductase inhibitor simvastatin. PMID- 17086618 TI - The value of temporal artery biopsy specimen length in the diagnosis of giant cell arteritis. PMID- 17086620 TI - Can pain be quantified numerically? PMID- 17086622 TI - Remitting seronegative symmetrical synovitis with pitting edema syndrome: followup for neoplasia. PMID- 17086624 TI - Cystic fibrosis: hope remains for a 'miracle cure'. PMID- 17086625 TI - The effects of marriage, civil union, and domestic partnership laws on the health and well-being of children. PMID- 17086626 TI - The effects of marriage, civil union, and domestic partnership laws on the health and well-being of children. PMID- 17086627 TI - The effects of marriage, civil union, and domestic partnership laws on the health and well-being of children. PMID- 17086628 TI - The effects of marriage, civil union, and domestic partnership laws on the health and well-being of children. PMID- 17086629 TI - [Understanding nephroblastoma. Information dedicated to parents and relatives]. AB - In response to the evolution of the information-seeking behaviour of patients and concerns from health professionals regarding cancer patient information, the French National Federation of Comprehensive Cancer Centres (FNCLCC) introduced, in 1998, an information and education program dedicated to patients and relatives, the SOR SAVOIR PATIENT program. The methodology of this program adheres to established quality criteria regarding the elaboration of patient information. Cancer patient information developed in this program is based on clinical practice guidelines produced by the FNCLCC and the twenty French regional cancer centres,the National League against Cancer, The National Cancer Institute, the French Hospital Federation, the National Oncology Federation of Regional and University Hospitals, the French Oncology Federation of General Hospitals, many learned societies, as well as an active participation of patients, former patients and caregivers. The handbook SOR SAVOIR PATIENT Understanding nephroblastoma is an adapted version of various scientific publications and international clinical practice guidelines, validated by oncology experts and by the Nephroblastoma Committee of the French Society against Cancers and Leukaemias in children and adolescents (SFCE). It was elaborated with the active participation of parents and other family members. It is meant to provide a basis for the explanation of the disease, to help parents asking questions, and to facilitate discussions with the healthcare team. It is available from the FNCLCC (101, rue de Tolbiac, 75013 PARIS, Tel. (0033)1 76 64 78 00, www.fnclcc.fr). This document was validated at the end of 2005 and published in May 2006. SOR SAVOIR PATIENT guides are systematically updated when new research becomes available. Information leaflets, extracted from the handbook SOR SAVOIR PATIENT Understanding nephroblastoma and published in this edition of the Cancer et Radiotherapie, describe the physiopathology of nephroblastoma, as well as treatments and follow-up. The guide allows parents and relatives to better understand the disease and the treatments proposed. It also offers health professionals a synthetic evidence-based patient information source which facilitates discussions with the patient. PMID- 17086630 TI - The National Institutes of Health: an institute on the move. A conversation with the director of the NIEHS. PMID- 17086631 TI - Chest X-ray quiz. Near drowning. PMID- 17086632 TI - Anaesthesia practitioners in the UK: the high road, not yet taken. PMID- 17086633 TI - How protected are coral reefs? PMID- 17086634 TI - How protected are coral reefs? PMID- 17086635 TI - How protected are coral reefs? PMID- 17086636 TI - Impact of nitrate supply in C and N assimilation in the parasitic plant Striga hermonthica (Del.) Benth (Scrophulariaceae) and its host Sorghum bicolor L. AB - The threshold of tolerance for nitrate of the parasitic weed Striga hermonthica (Del.) Benth and the host plant Sorghum bicolor L. was determined by estimating the impact of increasing nitrate loads on plant growth and various parameters of C and N assimilation. Nitrate supply improved chlorophyll (Chl) content and photosystem II (PSII) photochemistry of infected S. bicolor that, in comparison to S. hermonthica, displayed a low imbalance between C and N assimilation when nitrate was supplied up to 1500 mg N per plant. Indeed, nitrate supplies increased strongly the leaf N:C ratio of the parasite. The higher nitrate load induced strong accumulation of nitrate, nitrite and ammonium, and consequently the death of S. hermonthica. Nevertheless, lower nitrate loads (up to 500 mg N per S. bicolor in this study) promoted leaf expansion, PSII photochemistry and N metabolism of S. hermonthica mature (M) plants, as attested by the significant rise in soluble protein and free amino-acid contents. Following these N supplies, the nitrate tolerance of S. hermonthica was correlated with an increase in PSII activity and a high incorporation of N excess into asparagine. This confirmed the central role of asparagine in the N metabolism of S. hermonthica, although this detoxification pathway was insufficient to limit ammonium accumulation under higher nitrate loads. PMID- 17086637 TI - Milk ejection and milk flow pattern data needed for longer feeds. PMID- 17086638 TI - Response to review of the breastfeeding atlas. PMID- 17086639 TI - Examining the evidence: Knowing if and when to transfer a resident with pneumonia from the nursing home to the hospital or subacute unit. PMID- 17086640 TI - Research brief: Potential interventions focused on prevention of behavioral problems in nursing homes. PMID- 17086641 TI - Workmen's Circle Home and Infirmary for the Aged v. Fink. PMID- 17086642 TI - Delio v. Westchester County Medical Center. PMID- 17086643 TI - In re Kerr. PMID- 17086644 TI - Researchers uncover function of key Alzheimer's enzyme. PMID- 17086645 TI - Musical training improves brain development in children. PMID- 17086646 TI - Patient in a coma plays tennis. PMID- 17086647 TI - Allen Brain Atlas maps 21,000 genes of the mouse brain. PMID- 17086648 TI - Vogel v. Forman. PMID- 17086649 TI - Randolph v. City of New York. PMID- 17086650 TI - District 27 Community School. v. Board of Education. PMID- 17086651 TI - City of New York v. New St. Mark's Baths. PMID- 17086652 TI - Polio drops arouse suspicion in Uttar Pradesh. PMID- 17086653 TI - Transfusion-related HIV outbreak in Kazakhstan children. PMID- 17086654 TI - Action plans. PMID- 17086655 TI - USA makes HIV testing routine part of care. PMID- 17086656 TI - Extracellular matrix in lung disease. Proceedings of the Twenty-First Transatlantic Airway Conference. PMID- 17086657 TI - Open magnetic resonance imaging (MRI) scanners. AB - (1) In most MRI scanners, the patient examination table fits inside a long cylindrical tube. Large patients cannot be accommodated, and some persons experience claustrophobic reactions. Open MRI systems, in which the patient is placed between two plates, overcome these disadvantages. (2) Open MRI scanners are widely used in health care. High-field closed MRI systems are preferred for many examinations. (3) Early versions of open MRI scanners had low magnetic field strength, gave poorer image quality than most closed systems, and required longer examination times. Newer open scanners include machines with higher magnetic field strengths and improved image quality. (4) Closed high magnetic field scanners with short magnets and wide bore tubes offer improved comfort to patients, and may be an alternative to open scanners. (5) There is interest in using open systems for intra-operative and image-guided interventions. PMID- 17086658 TI - CFD investigation of room ventilation for improved operation of a downdraft table: novel concepts. AB - We report a computational fluid dynamics (CFD) study of containment of airborne hazardous materials in a ventilated room containing a downdraft table. Specifically, we investigated the containment of hazardous airborne material obtainable under a range of ventilation configurations. The desirable ventilation configuration should ensure excellent containment of the hazardous material released from the workspace above the downdraft table. However, increased airflow raises operation costs, so the airflow should be as low as feasible without compromising containment. The airflow is modeled using Reynolds Averaged Navier Stokes equations with a high Reynolds number k-epsilon turbulence model. CFD predictions are examined for several ventilation configurations. Based on this study, we find that substantial improvements in containment are possible concurrent with reduction in airflow, compared with the existing design of ventilation configuration. PMID- 17086659 TI - Pots, parties, and politics: communal feasting in the American Southwest. PMID- 17086660 TI - Doppler-guided hemorrhoidal artery ligation. PMID- 17086661 TI - Sandal types and Archaic prehistory on the Colorado Plateau. PMID- 17086662 TI - Myiasis: a perianal wound infection caused by fly larvae in gangrenous hemorrhoid. PMID- 17086663 TI - Metastasis in anal mucosa from bladder cancer. PMID- 17086664 TI - A comparison of total, respirable, and real-time airborne particulate sampling in horse barns. AB - Measurements of total, respirable, and real-time airborne particulate were obtained in 12 horse barns in summer and winter. Respirable and total particulate concentrations were measured gravimetrically; real-time particulate was measured with an aerosol photometer. Total particulate (TP) ranged from nondetectable (ND) to 2.1 mg/m3 and from ND to 1.2 mg/m3 for winter and summer sampling, respectively. Respirable particulate (RP) ranged from ND to 0.2 mg/m3 and from ND to 0.7 mg/m3 for winter and summer measurements, respectively. The mean respirable fraction of particulate for summer and winter measurements was 0.63 and 0.34, respectively Real-time particulate (RTP) concentration was significantly correlated with TP for winter measurements and for pooled data but was not significantly correlated with RP for winter, summer, or pooled measurements. Peak, 10-sec average levels of RTP were estimated to range up to 22 mg/m3 in the samples collected. The data suggest that (a) airborne particulate concentrations and particle size distributions vary considerably among horse barns, and (b) real-time sampling, with an aerosol photometer, may be a more reliable measure of airborne TP than airborne RP in these environments. PMID- 17086665 TI - Assessing occupational exposure to perchloroethylene in dry cleaning. AB - Occupational exposure to perchloroethylene (PERC) in dry cleaning is analyzed by calculating airborne concentrations for a large set of possible exposure scenarios. Different generations of dry-cleaning machines ranging from vented machines used until the 1980s to nonvented closed-loop machines used since the 1990s are investigated; the geographical scope of the study is Germany. Concentrations are calculated for diffuse emissions, release of contaminated air during loading and unloading, and residual solvent releases from cleaned garments with a dynamic two-box model for near field and far field. The concentration estimates for near field and far field agree well with measured concentrations. The airborne concentrations are compared with maximum workplace concentrations (MAK values). The full set of scenarios shows for which historical situations the workplace concentrations were near the MAK value and that the transition to newer dry-cleaning machines reduced the occupational exposure by more than one order of magnitude. In addition, numbers of exposed workers are estimated for different years. Whereas more than 25,700 workers in the near field were exposed to PERC in 1975, the number has been below 5900 since 2001, which is due mainly to a reduced amount of garments being dry cleaned and, to a lesser extent, to replacement of PERC by nonchlorinated solvents. The results from the scenario-based analysis are consistent with exposure ranges obtained from a Monte Carlo calculation using frequency distributions of key input parameters, such as emission factors, batch times, and room volumes. PMID- 17086666 TI - Powered, air-purifying particulate respirator filter penetration by a DOP aerosol. AB - In 1995, new certification requirements for all nonpowered, air-purifying particulate filter respirators were put in place when 42 CFR 84 replaced 30 CFR 11. However, the certification requirements for all other classes of respirators, including powered air-purifying respirators (PAPRs), were transferred to 42 CFR 84 from 30 CFR 11 without major changes. Since the inception of 42 CFR 84, researchers have learned that the efficiency of electrostatic filter media, in contrast with mechanical filter media, can be rapidly degraded by oil aerosols. Further, confusion may exist among respirator users, since electrostatic PAPR filters have the same magenta color assigned to high-efficiency filters for nonpowered particulate respirators that have been tested and certified for use against oil aerosols (i.e., P100 filters). Users may expect that the magenta color of certified PAPR filters indicates suitability for use against oil aerosols. This may not be the case. To illustrate the potential degradation of electrostatic PAPR filters, new filters certified under 42 CFR 84 were tested using a TSI model 8122 Automated Respirator Tester against charged and neutralized DOP aerosols with intermittent loading schedules. The performance of a magenta-colored electrostatic PAPR filter--one for which the manufacturer's user instructions appropriately indicates is not suitable for use in oily environments--was compared with the performance of several mechanical PAPR filters. In tests against both DOP aerosols, the electrostatic PAPR filter showed a significant decrease in performance at DOP loadings exceeding 400 mg, whereas mechanical filters showed no significant change in the performance except at extremely high loadings. The decreased performance of the electrostatic PAPR filter was found to be significantly greater when tested against a neutralized DOP aerosol when compared with a charged DOP aerosol. While laboratory tests show that the filtration efficiency of this electrostatic PAPR filter degrades with exposure to DOP aerosol, the observed laboratory degradation may or may not affect workplace performance, as similar degradation has not been verified in workplace studies. Based on these laboratory results, a proposed method for evaluating high-efficiency PAPR filters is presented. This proposed method would ensure that high-efficiency PAPR filters (> or = 99.97% efficient and magenta in color) meet critical performance criteria when loaded. PMID- 17086667 TI - More coverage, better care: improving children's access to health services. AB - As part of its continuing mission to serve trustees and staff of health foundations and corporate giving programs, Grantmakers In Health (GIH) convened a group of grantmakers and health services researchers on November 4, 2005 for an informative discussion about improving children's access to health services, including the opportunities, challenges, and roles for funders. This Issue Dialogue, entitled More Coverage, Better Care: Improving Children's Access to Health Services, covered how the current health care system succeeds and fails for children, emerging policy developments, and what grantmakers are currently doing to promote children's access to health services. The meeting also explored the lessons learned by grantmakers to help guide future work. PMID- 17086668 TI - Integrated unit performance testing of powered, air-purifying particulate respirators using a DOP challenge aerosol. AB - Although workplace protection factor (WPF) and simulated workplace protection factor (SWPF) studies provide useful information regarding the performance capabilities of powered air-purifying respirators (PAPRs) under certain workplace or simulated workplace conditions, some fail to address the issue of total PAPR unit performance over extended time. PAPR unit performance over time is of paramount importance in protecting worker health over the course of a work shift or at least for the recommended service lifetime of the PAPR battery pack, whichever is shorter. The need for PAPR unit performance testing has become even more important with the inception of 42 CFR 84 and the recent introduction of electrostatic respirator filter media into the PAPR market. This study was conducted to learn how current PAPRs certified by the National Institute for Occupational Safety and Health would perform under an 8-hour unit performance test similar to the dioctyl phthalate (DOP) loading test described in 42 CFR 84 for R- and P-series filters for nonpowered, air-purifying particulate respirators. In this study, entire PAPR units, four with mechanical filters and one with an electrostatic filter, were tested using a TSI Model 8122 Automated Respirator Tester, with and without the built-in breathing machine. The two, tight-fitting PAPRs, both with mechanical filters, showed little effect on performance resulting from the breathing machine. The two loose-fitting helmet PAPRs indicate that unit performance testing without the breathing machine is a more stringent test than testing with the breathing machine under the conditions used. The PAPR with a loose-fitting hood gave inconclusive results as to which testing condition is more stringent. The PAPR unit equipped with electrostatic filters gave the highest maximum penetration values during unit performance testing. PMID- 17086669 TI - Generalized P-values and confidence intervals: a novel approach for analyzing lognormally distributed exposure data. AB - The problem of assessing occupational exposure using the mean of a lognormal distribution is addressed. The novel concepts of generalized p-values and generalized confidence intervals are applied for testing hypotheses and computing confidence intervals for a lognormal mean. The proposed methods perform well, they are applicable to small sample sizes, and they are easy to implement. Power studies and sample size calculation are also discussed. Computational details and a source for the computer program are given. The procedures are also extended to compare two lognormal means and to make inference about a lognormal variance. In fact, our approach based on generalized p-values and generalized confidence intervals is easily adapted to deal with any parametric function involving one or two lognormal distributions. Several examples involving industrial exposure data are used to illustrate the methods. An added advantage of the generalized variables approach is the ease of computation and implementation. In fact, the procedures can be easily coded in a programming language for implementation. Furthermore, extensive numerical computations by the authors show that the results based on the generalized p-value approach are essentially equivalent to those based on the Land's method. We want to draw the attention of the industrial hygiene community to this accurate and unified methodology to deal with any parameter associated with the lognormal distribution. PMID- 17086670 TI - The path to policy change: practical steps and lessons from health funders. AB - As part of its continuing mission to serve trustees and staff of health foundations and corporate giving programs, on November 3, 2005, Grantmakers In Health (GIH) convened nearly 80 grantmakers and a diverse group of individuals with expertise in different types of public policy work to discuss the challenges and opportunities for health funders who seek to bring about change in federal, state, and local public policies. The program was designed to address funders' desire to go beyond the basics of funding public policy work and to learn from peers and others about how to improve the effectiveness of their public policy efforts. It was structured as a series of small-group discussions on key topics including advocacy infrastructure, communications, community organizing, data development and analysis, evaluation, working with foundation boards, and working with policymakers. PMID- 17086671 TI - Frederic Louis Lizzi. PMID- 17086672 TI - Prehistoric pipes from the Olds Ferry Dunes site (10-WN-557), Western Idaho. PMID- 17086673 TI - Early Paleoindian women, children, mobility, and fertility. PMID- 17086674 TI - Agnaiyaaq: the autopsy of a frozen Thule mummy. PMID- 17086675 TI - "Now dead I begin to sing": a protohistoric clothes-burning ceremonial feature in the Colorado Desert. PMID- 17086676 TI - Plastic shamans and astroturf sundances: New Age commercialization of Native American spirituality. PMID- 17086677 TI - Descriptive epidemiology of Lyme disease in Ontario: 1999-2004. PMID- 17086678 TI - Influenza outbreak in an Ontario long-term care home--January 2005. PMID- 17086679 TI - Spreading its wings. PMID- 17086680 TI - The imaging man. PMID- 17086681 TI - Reading the coffee beans. PMID- 17086682 TI - Neurofuturity: a theory of change. AB - Our experience is dominated by familiar sameness and unfamiliar change. This article is about change and why and how people change. The capacity to change is critical to the ability to adapt to a changing environment. Our brains help us to manage change by constructing possible futures and enabling us to behave in a way that prepares us for those futures. Capturing the world outside of us to make it into an internal experience is called perception. Some therapies aim to capture the work as it is. Capturing our world as it has been is termed 'memory'. Therapies which aim to revisit and remould this work so that the present and future are easier to deal with, help adaptation. Capturing the possible worlds to come is described here as neurofuturity. It is much broader than expectations and includes our feelings towards the future as well as our beliefs. Therapies are described in terms of which questions they answer and which time frame they address. Therapies and therapists address these big questions and work in different time frames. Reconstructing patients' experience of the future and acceptance of what cannot change are two of the main tasks of clinicians. PMID- 17086683 TI - Theories of change in therapeutic work. AB - This article explores how theories of personal change differ from each other according to the assumptions made about the nature of the individual, the relationship between the person and the social, and the nature of causality. Three different concepts of the individual person are distinguished, namely, the autonomous, the expressivist, and the social individual. Each of these implies different relationships between the individual and the social, and different theories of causality. From the perspective of the autonomous individual, change in a person is a rational reordering of individual thought processes. The cause of any personal change is rational effort of the individual. Clinical psychology and psychoanalysis reflect this perspective. From the expressivist perspective, each individual has an inner urge to self-actualize which is the cause of change. Humanistic psychology is based on this view. From the perspective of social individuals, the individual is a cultural being, necessarily dependent on others, who only develops a mind in interaction with others. From this perspective, individual change cannot be separated from change in the groups to which an individual belongs. PMID- 17086684 TI - An Eastern perspective on change. AB - In Eastern, particularly Buddhist, thinking the process of change and impermanence is an integral part of existence. All suffering in life is the result of ignorance of this basic reality. A deeper awareness and appreciation of this fact leads to enlightenment and real happiness. It is the rapidity of change that is responsible for the illusory processes that create the sense of solidity, continuity, permanence, entity, agency, self or ego. Thus, the mind is able to mix the real with the unreal. In reality, there are merely processes. However, mere intellectual understanding of this is not enough. It requires experiential understanding through the expansion of consciousness and attention to these processes. It also requires experiential training, appropriate attitudes (such as kindness, compassion, joy and equanimity) and development which is often gradual. Thus, learning to appreciate and delight in the existential process of change is at the heart of the Buddhist approach. It is beneficial not only in the long run (i.e., leading to enlightenment and freedom from the cycle of birth and rebirth, the ultimate purpose of life) but also in the short term (i.e., by making living and dying enjoyable). PMID- 17086685 TI - Pathways to recovery: promoting change within a developmental-systemic framework. AB - This article describes a format for the process of achieving therapeutic change through structured individual sessions with adolescents or adults with eating disorders. It is a model for change based on three separate existing theoretical or conceptual strands: Developmental theory; the application of systems theory and cybernetics to clinical practice; and feminist ideology. It was developed as a pragmatic, clinician-friendly model that could be successfully used by therapists from different disciplines, and is referred to here as developmental systemic-feminist therapy or individual developmental-systemic therapy. Change in this context is defined as the overt and measurable alteration in feelings, thoughts and behaviours of the participant(s) over the course of treatment. The article provides an overview of the treatment model and outlines the five steps used to achieve change (explore; understand; accept; challenge; change). Essential components of the therapist's stance, skills and knowledge are described, followed by a discussion of the structure and content of sessions in different stages of the therapy. Finally, clinical examples are given to illustrate the clinical use of this model in young patients with eating disorders. PMID- 17086686 TI - Mechanisms of action in the process of change: helping eating disorder clients make meaningful shifts in their lives. AB - This article describes a model for change that draws upon motivational and cognitive-behavioural theories. It is suggested that maladaptive beliefs hold individuals captive in destructive self-perpetuating cycles dominated by the illness. In the context of a trusting therapeutic relationship, inquiry and experimentation can lead to the reformulation of maladaptive beliefs, and to the development, reprioritization and/or strengthening of higher values. A clear cohesive system of higher values allows individuals to make autonomous life decisions, which reduces the need for maladaptive coping strategies. PMID- 17086687 TI - Something happens: elements of therapeutic change. AB - To make a difference therapists need a theory with which to understand the patient. They choose theories that suit them, and need technique to maintain the therapeutic setting. Psychotherapy developed from healing and medicine, and from our ethological heritage of care for dependants. It happens in a special setting which makes new experiences possible. In this article I list a number of ingredients that are necessary in any effective psychotherapy. PMID- 17086688 TI - Music therapy with sexually abused children. AB - Music is part of everyday life, and is generally regarded as therapeutic. There is increasing interdisciplinary interest in innate human musicality and the link between music and the emotions. Innate musicality is evident in the dynamic forms of emotional expression that both regulate and cultivate the foundations of meaning in human communication (intersubjectivity). This article discusses music therapy, drawing from interdisciplinary perspectives, and illustrated by case material of individual music therapy with a sexually abused child. Where the growth of mind and meaning is devastated at its core by early relational trauma, music, when used with clinical perception, may reach and work constructively with damaged children in an evolving, musically mediated therapeutic relationship. PMID- 17086689 TI - Art therapy for children: how it leads to change. AB - The aim of art therapy is to facilitate positive change through engagement with the therapist and the art materials in a safe environment. This article will explore how art therapy is used to help children with emotional, developmental and behavioural problems. It will show how change occurs during the process of physical involvement with the materials; through the making of a significant art object; through sublimation of feelings into the images; and through communication with the therapist via the art object. The article is illustrated with case vignettes which demonstrate how the theories underpinning art therapy are put into practice, drawing attention to the changes that occur as a result. PMID- 17086690 TI - The self-report of Andi. AB - This fictional account of a teenage girl's treatment is based on the author's experience of working with troubled young people. He shows how cases of self-harm and antisocial behaviour can be helped not only by an understanding of the intra psychic world, but by a therapeutic regime based on psychodynamic principles. In residential treatment there are opportunities for adolescents to learn from relationships with staff, peers and educational input but sometimes they are wasted by the repetition of abuse. While psychoanalytic psychotherapy by itself may be insufficient for a young person in need of a more comprehensive residential treatment, nevertheless psychodynamic concepts may make significant contributions to the process of therapeutic change. PMID- 17086691 TI - How and why parents change their attitudes to prenatal diagnosis. AB - Parents who know they are carriers of a genetically transmitted disease because they gave birth to an affected child are being offered a number of reproductive options in a subsequent pregnancy. We describe the attitudes and motives of parents of children with cystic fibrosis (CF) who were initially determined to use prenatal testing but eventually refused it. Based on our observations and clinical impressions we built a model of change that entails a series of stages and depicts change as a process. This begins with the provision of comprehensive information and emotional support through health professionals (Stage 1). The parents then engage in intense confrontation with the impact of prenatal testing and are exposed to influences from many sides. The repeated comparing of the arguments results in uncertainty and distress (Stage 2). When pregnancy occurs a decision must be made and the process culminates in participating in a life-or death decision. If the parents cannot cope with that responsibility then a change towards rejection of prenatal diagnosis is likely (Stage 3). This phenomenon seems to be the most decisive factor. A case vignette is used to illustrate this model which may be applicable to many individuals who face major and emotionally fraught decisions. PMID- 17086692 TI - A review of change: principles of problem formation and problem resolution. PMID- 17086693 TI - Castaway's corner. PMID- 17086694 TI - [Effects of mixed cypermethrin and methylparathion on endocrine hormone levels and immune functions in rats: II. Interaction]. AB - OBJECTIVE: To study interaction of mixed pesticides cypermethrin and methyl parathion on reproductive hormones, thyroid hormones, and immune functions in rats. METHODS: Eighty 2-month old Wistar rats (40 male and 40 female) were divided randomly by body weight into 8 groups. The dose 1/30 LD50 were chosen for the single or combined exposure representing respective doses of 0, cypermethrin 8.0 mg/kg bw, methylparathion 0.23 mg/kg bw, and 1/30 LD50 cypermethrin plus 1/30 LD50 methylparathion. The control group received vehicle solvent only. All groups were force-fed every two days for 30 days. Body weight gain and organ weights were determined. Serum levels of IgG and IgA, reproductive hormones [luteinizing hormone (LH), follicle stimulating hormone (FSH), estradiol (E2), and testosterone], as well as the thyroid hormones [triiodothyronine (T3), tetraiodothyronine (T4), and thyroid stimulating hormone (TSH) were measured using radioimmunoassay (RIA). In addition, two immunological parameters (rate of neutrophil phagocytosis, rate of lymphocyte transformation] were being measured in blood samples. RESULTS: The most of index indicated addictive interaction, while the effects on relative weights of ovaries and adrenals, IgA and rate of lymphocyte transformation were antagonistic. It was of interest that the effect on estradiol was synergistic interaction in female rats, whereas it was addictive interaction in male rats, whose estradiol level could be increased 64.64% by cypermethrin exposure. CONCLUSION: Our results showed that exposure to cypermethrin and methyl parathion mixture at 1/30 LD50 dose had interaction on endocrine hormone levels, and immune functions in rats. Estradiol was very sensitive, the mixture can enhance estradiol level both in male and female rats. PMID- 17086695 TI - [Polymorphisms of the DNA repair genes XRCC1 and XPC: relationship to pancreatic cancer risk]. AB - OBJECTIVE: To determine whether genetic polymorphisms in XRCC1 and XPC are associated with risk of pancreatic cancer. METHODS: A case-control study was conducted in 101 incident cases with pancreatic cancer and 337 controls (matched for age, sex and ethnicity) to investigate whether genetic polymorphisms in DNA repair genes XRCC1 (Arg194Trp and Arg399Gln) and XPC (an intronic biallelic poly (AT) insertion/deletion polymorphism, XPC-PAT) were associated with risk of pancreatic cancer. The odds ratios (ORs) and their 95% confidence intervals (CIs) were calculated by unconditional logistic regression models and adjusted for potential confounding factors. RESULTS: There was a small, non-significant decrease in risk for pancreatic cancer in those carrying Gln/Gln genotype at XRCC1 Arg399Gln site (OR 0.64, 95% CI 0.21 - 1.66, P = 0.30) compared with those having Arg/Arg genotypes. And the XRCC1 Arg194Trp polymorphism was not significantly associated with risk of pancreatic cancer. For XPC-PAT polymorphism, 5.0% of cases and 13.4% of controls were homozygous for the variant allele (PAT+/+), resulting in an OR of 0.30 (95% CI 0.10 - 0.76, P = 0.02), which suggested that the PAT +/+ genotype might have protective effect against pancreatic carcinogenesis. CONCLUSIONS: This study suggest that XPC-PAT polymorphisms may contribute to the risk of pancreatic cancer in our study population. PMID- 17086696 TI - [Effect of ceramide on apoptosis of human colon cancer HT-29 cells]. AB - OBJECTIVE: To investigate the effect of apoptosis and mRNA expression of apoptosis-related genes of human colon cancer HT-29 cells induced by exogenous ceramide. METHODS: After HT-29 cells exposed to C2-ceramide for different time, transmission electron microscope and fluorescence microscope were used to observe the morphology change of cells. DNA agarose gel electrophoresis was applied to detect DNA fragmentation of HT-29 cells. The mRNA expression of apoptosis-related genes was determined by reverse transcription polymerase chain reaction (RT-PCR) analysis. RESULTS: Under the contribution of C2-ceramide, typical apoptosis appearance, such as nuclear chromatin breakage, apoptotic body etc and DNA ladder were observed. Exposing to 50 micromol/L C2-ceramide for 12h and 24h, the rate of apoptotic cells were 64.7% and 81.3% respectively, which appeared time-dose relation. Meanwhile, ceramide increased the expression of Bax, Bad and Bid mRNA, and decreased the expression of Bcl-2 and Bcl-xl mRNA. CONCLUSION: Ceramide may induce apoptosis of human colon cancer HT-29 cells via up-regulating or down regulating the expression of apoptosis-related genes. PMID- 17086697 TI - [Construction of thr461 --> Asn461 and Ile462 --> Val462 mutation vector of P4501A1 gene]. AB - OBJECTIVE: To construct Thr461 --> Asn461 and Ile462 --> Val462 mutation vector of P4501A1 gene and to provide scientific base for deeply researching on the function of cytochrome 1A1 gene (CYP1A1) and the mechanism of carcinogenesis. METHODS: According to cDNA sequence of human CYP1A1 gene, universal primers (Pm3/Pm4) and mutant primers (Pt15/Pt16 and Pt17/Pt18) containing restriction enzyme site and mutation site were designed. The first set of primers involving Pm3/Pt16 and Pm3/Pt18 amplified a forward 1.5kb fragment from pGEM-T-CYP1A1 plasmid. The second set of primers involving Pt15/Pm4 and Pt17/Pm4 amplified a reverse 177-bp fragment from 10ng pGEM-T-CYP1A1 plasmid. The third set of primers involving Pm3/Pm4 amplified a 1.5kb fragment from the fomer PCR amplifications. The third PCR products were separated, purified and recovered from 1% agarose gel, then inserted into pMD-T vector. Subsequently the conjunct products were transformed into E. coil strain DH-5alpha., then the single clone was screened out and plasmids were extracted from such clone finally verified by restriction endonuclease analysis and sequencing. RESULTS: A 1.5kb fragment of tricycle PCR amplifications were digested by restriction endonucleases (BamHI and SailI) and sequenced bidirectionally by universal primers(T7p and SP6). The results verified that the cloned fragment including Asn461 and Val462 mutant site had 99.9% homology with the human cDNA of CYP1A1 gene in Genebank. CONCLUSION: The objective fragment containing Asn461 and Va462 mutant site with cDNA of the CYP1A1 gene has been successfully constructed in this experiment. PMID- 17086698 TI - [Study of dicyclohexyl phthalate on preparation and characterization of artificial antigen]. AB - OBJECTIVE: To study the preparation and characterization of hapten and artificial antigen of dicyclohexyl phthalate. METHODS: A dicyclohexyl phthalate hapten (4 amino dicyclohexyl phthalate) was synthesized by introducing amino as a substituent on the aromatic ring and retaining the ester group, and characterized by 1HNMR, IR and UV. The hapten was conjugated to BSA via amino diazotization linkage. RESULTS: Lambda1 = 214nm, lambda2 = 256nm for the UV of dicyclohexyl 4 nitrophthalate and lambda1 = 226nm, lambda2 = 288nm for the UV of dicyclohexyl 4 aminophthalate. Artificial antigen was prepared and tested by fluorescence, and lambda(ex) = 307nm, lambda(em) = 468nm, and the approximate molar ratio of dicyclohexyl 4-aminophthalate to BSA was 19. The product was used as an immunogen, demonstrating that it is suitable for polyclonal antibody production. CONCLUSION: It is a good method for preparation of artificial antigen of dicyclohexyl phthalate by introducing amino as a substituent on the aromatic ring and retaining the ester group. It was suggested that could supply excellent immune antigen for further preparation of antibody and immunoassay to dicyclohexyl phthalate. PMID- 17086699 TI - [Research in the relation between telomerase reverse transcriptase expression in spermatogenic cells and serum levels of estradiol of fluorotic rats]. AB - OBJECTIVE: To study the relation between telomerase reverse transcriptase (TERT) activity expression in spermatogenic cells and serum levels of estradiol of fluorotic rats. METHODS: We randomly divided thirty SD male rats into control group, low-dose group and high-dose group, then inject sodium fluoride (0, 10, 20 mg/kg bw) into celiac of rats. We respectively observed changes of estrogen and TERT using methods of radioimmunoassay, in situ hybridization. In addition, we observed the quality of spermatozoa. RESULTS: The level of estrogen, the expression of telomerase and the number and the livability of the spermatozoon in low-dose and high-dose fluorotic rats were lower than those of control rats (P < 0.05). Therefore,the above indexes decreased with the increase of dosage. In addition, sperm aberration of each fluorotic group was higher than control group (P < 0.05). And it increased with the increase of dosage. The content of E2 in serum of different fluoride treatment groups was positively correlated with the expression of telomerase in seminiferous tubule significantly, respectively (low dose fluoride treatment groups, r = 0.941, P < 0.01, high-dose fluoride treatment groups, r = 0.929, P < 0.01). CONCLUSION: NaF possibly damaged the male reproductive system by the approach of E2/ER-TERT-spermatozoon, relation between TERT expression in spermatogenic cells and serum levels of estradiol is positive correlation. PMID- 17086700 TI - [Comparative study of nanosized and microsized silicon dioxide on spermatogenesis function of male rats]. AB - OBJECTIVE: To compare the effects of nanosized and microsized silicon dioxide on spermatogenesis function of male rats exposed by inhalation. METHODS: 45 male rats were randomly divided into control group and four experimental groups which were exposed by 100 mg/m3 or 300 mg/m3 nanosized and microsized silicon dioxide in inhalation chambers 2 hours every other day. Age-matched rats were exposed to room air with the same condition and served as controls. 65 days later, the testicular and epididymal viscera coefficients, the quantity and quality of sperm were examined and the histopathological assessment was done. The changes in biochemical parameters in serum and testes were also measured. RESULTS: Nanosized silicon dioxide could induce histopathological changes of testes in rats, and the effect was higher than that of microsized particles at the same concentration. Nanosized silicon dioxide could reduce the sperm counts of rats and the testicular LDH-C4 activities, increase MDA levels in the testes and the effect was higher than that of microsized particles at the same concentration. Nanosized silicon dioxide could lead to the reduction of sperm motility, testicular LDH-C4 activities and 8-hydroxydeoxyguanosine (8-OHdG) concentration in serum elevation in particles-exposed rats compared with the control animals, but there are no significant difference compared with that of microsized particles at the same concentration. CONCLUSION: The present findings suggest a different effect of impairment of sperm production and maturation induced by inhalation of nanosized and microsized silicon dioxide, and nanosized silicon dioxide exerted more severe reaction. PMID- 17086701 TI - [Effects of sulfur dioxide on mRNA expression and activities of hepatic and pulmonary cytochrome P4502B1 and 2E1 in rats]. AB - OBJECTIVE: To study the effects of sulfur dioxide (SO2) on activities and mRNA expression of hepatic and pulmonary cytochrome P4502B1 and 2E1 in rats. METHODS: Male Wistar rats were housed in exposure chambers and treated with 14, 28 and 56 mg/m3 SO2 for 6 h/d for 7 days, while control rats were exposed to filtered air in the same condition. The activities of CYP2E1 of rats were measured by spectrophotometry. Fluorescence spectrophotometry was used to study the activities of CYP2B1. The mRNAs of CYP2B1 and 2E1 were analyzed in livers and lungs by using a reverse transcription-polymerase chain reaction (RT-PCR) assay. RESULTS: In the liver, decreases of CYP2B1 activity and mRNA were observed at higher dose of SO2 (28 and 56 mg/m3). However, the CYP2E1-dependent p-nitrophenol hydroxylases (p-NP) and mRNA were unaltered by SO2 at all detected concentrations. For lungs, CYP2B1 activity was unaltered by SO2 at low concentrations, except for a significant decrease in the rats exposed to SO2 at 56mg/m3, however, SO2 at higher concentrations (28 and 56 mg/m3) significantly decreased CYP2B1 mRNA. Significant inhibition of p-NP was observed in lungs of rats exposed to SO2 at 28 and 56 mg/m3. SO2 at higher concentrations (28 and 56 mg/m3) decreased significantly pulmonary CYP2E1 mRNA relative to control animals. CONCLUSION: It was suggested that SO2 exposure could suppress the activities and mRNA expression of hepatic CYP2B1 and pulmonary CYP2B1 and 2E1 of rats. PMID- 17086702 TI - [Effects of the inhalable particle (PM10) on secretion of inflammatory factors in human lung fibroblasts and mouse alveolar macrophage cell]. AB - OBJECTIVE: To study the effects of PM10 and the concentrated culture suspension of RAW264.7 cells treated with PM10 on the secretion of tumor necrosis factor alpha (TNF-alpha), interleukin-6(IL-6) and interleukin-8(IL-8) in the human lung fibroblasts( HLF). METHODS: PM10 were collected during heating period in the urban area of Beijing. HLF and mouse alveolar macrophage cell (RAW264.7) were used. The two ways of exposed cells to PM10 were used: (1) HLF were treated with the different concentration of PM10 for 24h. (2) HLF were stimulated by the added the suspension of RAW264.7 cell exposed to different concentrations of PM10. Cytotoxicity of PM10 was measured by MTT assay. The levels of inflammatory cytokines TNF-alpha, IL-6 and IL-8 were determined by the radioimmunity assay. RESULTS: After treated for 24h, PM10 showed the cytotoxicity in HLF and RAW264.7 cells, which was characterized by increase of the viability of cells at low concentration of PM10, and decrease at high doses. PM10 induced the secretion of TNF-alpha, IL-6 and IL-8 in HLF in a dose-dependent manner. The concentrated culture medium of RAW264.7 cells treated with PM10 also stimulated the secretion of TNF-alpha, IL-6 and IL-8 in HLF. CONCLUSION: PM10 is cytotoxic to HLF and RAW264.7 cells. PM10 induced the secretion of TNF-alpha, IL-6 and IL-8 in HLF. This effect was also observed by treatment of HLF with the concentrated culture medium of RAW264.7 cell exposed to PM10. PMID- 17086703 TI - [Comparison of perfluorooctane sulfonate and perfluorooctane acid in serum of non occupational human from Shenyang and Chongqing areas]. AB - OBJECTIVE: The purpose of this investigation was to illustrate the perfluorooctane sulfonate (PFOS) and perfluorooctane acid (PFOA) levels in serum of non-occupational exposure human from Shenyang and Chongqing areas and to compare the distributing character and region difference of PFOS and PFOA in those two region human. METHODS: Sera samples of non-occupational human from Shenyang and Chongqing areas were collected, and the concentration of PFOS and PFOA in serum were measured by High Performance Liquid Chromatography/Mass Selective Detector (HPLC/MS-MIS). RESULTS: The average Shenyang and Chongqing fluorochemical concentrations, respectively, were as follows: PFOS, 22.40 microg/L vs 7.40 microg/L, PFOA, 4.32 microg/L vs 1.00 microg/L. Statistical analysis indicated that serum concentrations of PFOS and PFOA were significantly (P < 0.01) higher in Shenyang human than in Chongqing human. Furthermore, there are sex differences in PFOS and PFOA concentrations in serum at all location. In Shenyang the concentration of PFOS in females were significantly (P < 0.05) higher than in males. The correlations of PFOS (r = 0.298) and PFOA (r = 0.271) with age were significant in females from Chongqing area, and especially the correlations were higher in older females (age t 50) than the groups of age < 13 and 13 - 50 years old. CONCLUSION: This finding suggests that there are predominant regional differences and distributing character for both PFOS and PFOA concentrations in Shenyang and Chongqing areas, and the concentrations of PFOS and PFOA in serum were correlated with age and sex. PMID- 17086704 TI - [Targeting hTERT gene influenced the viability on hMSCs by small hairpin RNA]. AB - OBJECTIVE: To observe the effect of shRNA by targeting human telomerase reverse transcriptase (hTERT) mRNA in the human mesenchymal stem cells (hMSCs) lines and explore the safety of RNAi by targeting hTERT mRNA in clinical application. METHODS: According to the strategy of RNAi, which determined the specific base sequences to design shRNA plasmid. Plasmid shRNA1 involved in fluorescein gene and homologous to hTERT were designed. control shRNA2-a random sequences heterogeneous to mankind's gene were also constructed. METAFECTENE was used as the transfection reagent. The experiment was divided into 4 groups: A (shRNA1), B (shRNA2), C (metafectene) and D (normal culture medium ). At 24h, fluorescence expression was detected by confocal microscopy after administration of plasmid. At 24h, 48h and 72h cells viability were determined using the MTT assay. At 48h, the morphological change were examined by HE stain. At 48h, hTERT mRNA were detected by reverse transcription polymerase chain reaction (RT-PCR) and the telomerase activity of hMSCs were examined by PCR ELISA. RESULTS: Many cells gave out green fluorescent after treated by shRNA1 and shRNA2. It was observed that the viability of hMSCs hadn't change after treatment with different methods within 72h (P > 0.05). Cells, morphology and density had not change after treated by different factor. All cells didn't express hTERT mRNA and the telomere activity of hMSCs was negative. CONCLUSIONS: The results suggest that shRNA plasmid directed against hTERT can not influence the viability on hMSCs. The treatment with RNA interference may be a safety strategy for gene therapy in vivo. PMID- 17086705 TI - [Effects of grape procyanidins on the concentration of intracellular calcium and the proliferation activity of the hepatoma cells]. AB - OBJECTIVE: To investigate the effects of grape procyanidins (GPC) on concentration of intracellular calcium and the proliferation activity of normal hepatic cells and the hepatic cell injuried by alcohol. METHODS: Rat hepatic cells and the cell injuried by alcohol were cultured with different concentration of GPC. The proliferation activity and concentration of intracellular calcium of the hepatic cells were measured by MTT assay and Fura-2 fluorescence methods. RESULTS: (1) The concentration of intracellular calcium of the normal control group and alcohol injury group were (108.26 +/- 14.17) and (651.24 +/- 47.95) nmol/L respectively, and that of both the high and the medium dose GPC groups were lower than the alcohol injury group, all differences are significant (P < 0.05). (2) The concentration of intracellular calcium in the normal hepatic cells treated with GPC is the group with calcium of extracellular fluid > the group free from calcium of extracellular fluid > the normal control group. (3) The proliferation activity of the high and the medium dose GPC group of normal hepatic cell and the cell injuried by alcohol were higher than the normal control group and alcohol injury group, all differences are significant (P < 0.05). CONCLUSION: GPC can raise the intracellular calcium concentration of normal hepatic cell by increasing extracellular fluidca introaffluxion and release from calcium pool, and enhance the proliferation activity of hepatic cells. It also can inhibit the abnormal rise (overload) of intracellular calcium concentration and the proliferation activity injury of hepatic cell induced by alcohol. PMID- 17086706 TI - [Effect of different post-weaning dietary compositions on body fat content and hormone-sensitive lipase gene expression in rats fed with high-fat diet]. AB - OBJECTIVE: To study the effects of different postweaning dietary compositions on body fat content and hormone-sensitive lipase (HSL) gene expression in rats fed a high-fat diet. METHODS: Male Wistar rats weaned on 24 days were randomly divided into A, B, C, D groups and were fed on high carbohydrate, high protein, high unsaturated fatty acid and high saturated fatty acid diet respectively. After 3 weeks, all rats were assigned to basic diet for two weeks, then rats from A group were randomly divided into A1 and A2 subgroups. A1 subgroup was fed on basic diet continuously. A2 subgroup and B, C and D groups were assigned to high-fat diet for six weeks. Eight rats from each group were sacrificed at the end of different treatments to detect body weight, body fat content and blood glucose level. RT PCR was used to determine the gene expression of hormone-sensitive lipase in white adipose tissue. RESULTS: Body weight, body fat content and blood glucose level in C group were significantly lower than those in A2 subgroup on 11 weeks (P < 0.05). The body weight, body fat content in B2 group were significantly lower than those in A2 subgroup (P < 0.05), but blood glucose level was higher than A2 subgroup on 11 weeks (P > 0.05). The body weight in D group was significantly lower than that in A2 subgroup (P < 0.05), but body fat content and blood glucose level were not significantly lower than A2 subgroup on 11 weeks (P > 0.05). The expression of HSL in B2 and C group was higher on 9 weeks and 11 weeks respectively. The HSL mRNA levels in all groups on 3 weeks were similar to that on 11 weeks. CONCLUSION: Feeding on high unsaturated-fatty acid diet after weaning may significantly reduce the body weight, body fat content, improve the abnormal blood glucose level in rats induced by high-fat diet. It was suggested that the high unsaturated-fatty acid diet can be reasonable for weaned rats and gene expression regulation may be involved in programming later obesity by early nutrition. PMID- 17086707 TI - [Survey on the prevalence of hypertension in different ethnic groups in China in 2002]. AB - OBJECTIVE: To understand the prevalence of hypertension and blood pressure level in different ethnic groups in Chinese population. METHODS: Data are presented for 67700 men and 84983 women aged more than 15 years old from National Nutrition and Health Survey, 2002. RESULTS: After being adjusted by age using Chinese population in 1964, the highest prevalence of hypertension in the male were. Tibetan (25.6%) and Manchu (23.1%). The Miao (9.2%) and Tujia (11.1%) ethnics have the lowest prevalence of hypertension. Also prevalence rate of hypertension in female in Tibetan (24.0%) and Manchu (18.7%) were the highest, and the lowest were Miao (6.1%) and Zhuang (8.3%) ethnics. The prevalence rate of hypertension in most ethnics were increased compared with that of 1991, except that of the male in Mongolian and female in Miao ethnics. CONCLUSION: The prevalence of hypertension and blood pressure level were different among ethnics. The prevalence rate of hypertension in almost all ethnics were increased during past ten years. PMID- 17086708 TI - [Association of polymorphism of low density lipoprotein receptor-related protein 5 Q89R, A1330V with bone mineral density in premenopausal northern Chinese women]. AB - OBJECTIVE: To investigate the association of lipoprotein receptor-related protein 5 (LRP5) gene Q89R, A1330V polymorphism with bone mineral density (BMD) in premenopausal Northern Chinese women. METHODS: 283 unrelated healthy residents of Han nationality in Harbin, aged 20 - 50, were selected. All subjects completed a food frequency questionnaire administered by a trained dietitian to estimate dietary calcium intakes, and were measured values of biochemical and hormone, such as the level of serum calcium, serum phosphorus, alkaline phosphatase, parathyroid hormone, calcitonin, 25 (OH) D3, 1, 25 (OH)2D3. The BMD phenotypes were measured by dual energy X-ray densitometry. The polymorphisms of LRP5 gene were determined using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). RESULTS: Frequencies of LRP5 genotype QQ, QR, AA and AV were 83.4%, 16.6%, 64.3% and 35.7% in all subjects. It failed to find that the values of serum biochemistry and the levels of hormones were statistically different among genotypes. Stepwise multiple regression analysis showed that the strongest predictor for BMD was body weight, other significant predictors were age and LRP5 genotypes, A1330V polymorphism were associated with BMD at lumbar spine, however Q89R polymorphism were associated with BMD at shaft and total hip (P = 0.033 and 0.034 respectively). CONCLUSION: LRP gene polymorphism was predictor for BMD in Northern Chinese premenopausal women, BMD at various were modified by different genetic and lifestyle factors in female. PMID- 17086709 TI - [Effects of phytosterol ester on serum lipid level in rats]. AB - OBJECTIVE: To observed the effects of phytosterol ester (PSE) on serum lipid level in rats. METHODS: (1) Under high-fat fed model, rats were oral administrated either no PSE (model), 4.0, 16.0 or 32.0 mg/kg bw PSE dissolved in oil. After 30 days of this treatment serum was obtained for lipid determination. (2) Ovariectomized rats were randomly assigned to model group, or treated with diethyl stilbestrol (E2, 22.5 microg/kg bw), PSE (32.0 mg/kg bw) or free phytosterol (FPE, 500 mg/kg bw). After fed for 60 days, body weight, liver and uterus weight, serum lipid and estrogen level were measured. In both experiments, another group of normal rats were taken as control. RESULTS: High-fat fed rats administrated with 3 doses of PSE had significantly lower triglyceride (TG) following 30 days of treatment compared to model group. A significant decrease in total cholesterol (TC) and low density lipoprotein cholesterol (LDL-C) was observed in 32.0 mg/kg bw PSE group, while a significant increase in high density lipoprotein cholesterol (HDL-C) and ratio of HDL-C/TC was observed in the same group ( P < 0.05). For ovariectomized rats treated with PSE or FPE, a significant lower body and liver weight with lower TC and TG level were observed in comparison with model group (P < 0.05). A little increased uterus weight by PSE and FPE was also observed, though no significant elevation in estrogen was reported. CONCLUSION: Phytosterol ester have potential to decrease hyperlipidemia. PMID- 17086710 TI - [Determination of galactooligosaccharide in syrups by high performance ion exchange chromatography method]. AB - OBJECTIVE: The gradient procedure was developed based on AOAC 2001.02 to determine galactooligosaccharide (soluble dietary fiber with low molecular weight) by high performance ion exchange chromatography with pulsed amperometric detection. METHODS: In the present study, the modified gradient procedure was used to separate the mixed standard on a CarboPac PA1 (2mm x 250mm) anion exchange column. The galactooligosaccharide in syrups were quantified and the repeatability and reproducibility of the modified method were also evaluated. RESULTS: Galactose, glucose, sucrose, fructose and lactose were successfully separated. The galacto oligosaccharides contents in syrups of intra-assay were (49.59 +/- 3.34), (46.55 +/- 2.61) and (56.26 +/- 3.13)g/100g, RSD value were 6.73%, 5.62% and 5.56% respectively. RSD value for galactooligosaccharide of inter-assay in syrup 1 was 4.73%. CONCLUSIONS: Galactose and glucose were separated well and galactose was determined more accurately. the repeatability and the reproducibility of the modified method were excellent. This method can be used for GOS determination. PMID- 17086711 TI - [Effect of probiotics and yogurt on colonic microflora in subjects with lactose intolerance]. AB - OBJECTIVE: To study the effects of probiotics and yoghurt supplementation on colonic microflora in subjects with lactose intolerance (LI). METHODS: Eleven lactose intolerant volunteers determined by 25g lactose challenge test were recruited and given supplementation with Bifina capsule (with B. longum) and yogurt (with B. animalis, L. bulgaricus, S. thermophilus) for 14 days. The trial was divided into 3 phases: 7-day basal period, 14-day supplementation period and 7-day post-supplementation period. Two lactose challenge tests were performed 1 day before and 1 day after supplementation period respectively, and lactose intolerance symptoms were recorded with questionnaires. Five fecal samples were collected during the trial, and total bacteria and several main groups of bacteria in colon were analyzed by fluorescence in situ hybridization (FISH). The shift of Bifidobacterium species population in colon was analyzed by denaturing gradient gel electrophoresis (DGGE). RESULTS: It was found that LI symptom was alleviated significantly after supplementation (P < 0.05). The amount of total bacteria and the main groups of bacteria (Bifidobacteria, Bacteroides-Prevotella, Eubacteria, Ruminccocc, et al.) was significantly increased during the supplementation period (P < 0.05). The amount of Bifidobacteria during post supplementation period was significantly higher than that of basal period (P < 0.05), while the amount of other several groups of bacteria decreased. The result of DGGE showed that the extraneous Bifidobacteria species from Bifina capsule and yoghurt could be detected in fecal samples during supplementation period, however there was diversity between volunteers. CONCLUSION: It was suggested that extraneous Bifidobacteria in Bifina capsule and yogurt could resist the effect of gastric acid and bile salt, and survive in colon. Supplementation of probiotics and yogurt could influence the colonic bacteria, and increase the number of main groups of colonic microflora, especially for Bifidobacteria, which could play important role in alleviating LI symptoms. PMID- 17086712 TI - [Rapid detection of influenza virus A by fluorescence quantitative RT-PCR]. AB - OBJECTIVE: To develop a method to rapidly detect influenza virus type A by fluorescence real-time quantitative RT-PCR. METHODS: According to conservative sequence of influenza virus type A M2 gene, a pair primers and Taqman probe were designed, respectively. After the quantitative curve of the assay were established using tenfold serial dilution of TCID50, the sensitivity and the specificity of clinic samples were determined. RESULTS: The detection sensitivity of the assay was 2.56 x 10(-5) TCID50 and the regression coefficient of the quantitative curve was 0.999. The specificity was determined by testing five other specimens, all of which yielded negative results. CONCLUSION: The detection system based on real-time RT-PCR was rapid, sensitive and steady, which could be used to detect the clinic samples. PMID- 17086713 TI - [Study on the occupational stress norm and it's application for the marketing group, public service/safety group and production laborer group]. AB - OBJECTIVE: A study of the occupational stress norm and it's application for the marketing group, public service/safety group and production laborer group. METHODS: In this study, cross-sectional study method is used, and a synthetic way of sorting and randomized sampling is adopted to deal with research targets (36 marketing group, 331 public service/safety group, 903 production laborer group). RESULTS: Descriptive statistics for OSI-R scale scores for the marketing group, public service/safety group and production laborer group were modulated. Scale raw score to T-score conversion tables derived from the OSI-R normative sample for marketing group public service/safety group and production laborer group were established. OSI-R profile from for marketing group, public service/safety group and production laborer group were established. For the ORQ and PSQ scales, scores at or above 70 indicate a strong levels of maladaptive stress and strain. Score in the range of 60 to 69 suggest middle levels of maladaptive stress and strain. Score in the range of 40 to 59 indicate normal levels of stress and strain. Score below 40 indicate a relative absence of occupational stress and strain. For the PRQ scales, score below 30 indicate a significant lack of coping resources. Score in the range of 30 to 39 suggest middle deficits in coping resources. Score in the range of 40 to 59 indicate average coping resources. Scores at or above 60 indicate a strong levels of coping resources. CONCLUSION: The authors combined subjective and objective environment match model of occupational stress. Different intervention measure should be take to reduce the occupational stress so as to improve the work ability. PMID- 17086714 TI - [Study on short-term therapeutic effect and its impact factors of stereotactic surgery for treating opiate users with opiate dependence in Sichuan]. AB - OBJECTIVE: To investigate the common characteristics of the patients, relapse reasons of patients before and after surgery, evaluate the relapse rate and its impact factors, and therapeutic effect of patients accepted stereotactic surgery to treat opiate dependence in Sichuan. METHODS: An investigation, using uniform questionnaires by face-to-face and telephone interview, and gaining data from medical records of patients, was conducted in Mar to Jun 2005, in Sichuan Province. 208 patients (total 212 patients participated in surgery) were invited to gather information about their common characteristics, drug-taking history and the surgery. Statistical methods including t-test, chi2 test and logistic regression were used to analyze the data. RESULTS: (1) 181 male patients and 27 female patients participated in this study, and their mean age was (29.5 +/- 5.5) years. Most of the respondents were in Sichuan and some peripheral province, graduated from senior high school and over, and with various occupations. (2) All patients abused opiate before the surgery, and the average duration of drug taking was (7.6 +/- 3.4) years. All patients were detoxified by unconstraint or compulsory abstinence before surgery, with mean drug abstinence of 13.9, but relapse occurred after each detoxification. (3) Suffering with no drugs and abstinence syndrome were the two main reasons of relapse before surgery. Compared with relapse before surgery, validating the effect of the surgery treating drug dependence and temptation by drug surroundings were the two main reasons of relapse after surgery. (4) The complication rate was 38.0% (79/208), no severe complications occurred in patients, and most of the complications disappeared or were healed before they were discharged from hospital. Relapse rate within 7 months after surgery was 22.1% (46/208). A significant decrease of relapse time, relapse dose, subjective feel on drugs and relapse euphoria appeared in patients who relapsed after surgery when compared with those before surgery. The univariate and multivariate analysis shows potential significant predictors of relapse rate after surgery to include education (OR = 3.259), operative time (OR = 2.451), social support (OR = 23.256) and doing simple work (OR = 3.328). CONCLUSION: This investigation showed that the stereotactic surgery can eliminate psychological desire for drugs and abstinence syndrome among most of the patients. Satisfactorily short-term therapeutic effect and substantial decline in relapse rate as well as no severe complications were appeared in these patients. Relapse was greatly associated with education, operative time, neuropsychological factors, and social conditions of patients. Therefore, patients' family and the society should strengthen their care, comprehension as well as support, and create better living and working surroundings to facilitate the complete drug abstinence to occur in patients. average duration of drug-taking was (7.6 +/- 3.4) years. All patients were detoxified by unconstraint or compulsory abstinence before surgery, with mean drug abstinence of 13.9, but relapse occurred after each detoxification. (3) Suffering with no drugs and abstinence syndrome were the two main reasons of relapse before surgery. Compared with relapse before surgery, validating the effect of the surgery treating drug dependence and temptation by drug surroundings were the two main reasons of relapse after surgery. (4) The complication rate was 38.0% (79/208), no severe complications occurred in patients, and most of the complications disappeared or were healed before they were discharged from hospital. Relapse rate within 7 months after surgery was 22.1% (46/208). A significant decrease of relapse time, relapse dose, subjective feel on drugs and relapse euphoria appeared in patients who relapsed after surgery when compared with those before surgery. The univariate and multivariate analysis shows potential significant predictors of relapse rate after surgery to include education (OR = 3.259), operative time (OR = 2.451), social support (OR = 23.256) and doing simple work (OR = 3.328). CONCLUSION: This investigation showed that the stereotactic surgery can eliminate psychological desire for drugs and abstinence syndrome among most of the patients. Satisfactorily short-term therapeutic effect and substantial decline in relapse rate as well as no severe complications were appeared in these patients. Relapse was greatly associated with education, operative time, neuropsycological factors, and social conditions of patients. Therefore, patients' family and the society should strengthen their care, comprehension as well as support, and create better living and working surroundings to facilitate the complete drug abstinence to occur in patients. PMID- 17086715 TI - [PCBs concentrations in human milk collected from a polluted area--exposure risk for feeding infants]. AB - OBJECTIVE: To determine the concentration of PCBs and the distribution of dioxin like compounds in human milk collected from a polluted area where discarded electronic equipment is dismantled, and to evaluate the risk for feeding infants. METHODS: The authors collected 20 and 12 milk samples from the polluted area and nearby town, respectively. Milk samples were analyzed for total PCBs (t-PCBs) by gas chromatography, and two mixed milk samples from the two areas were detected the distribution of dioxin-like compounds. RESULTS: t-PCBs in human milks of polluted area and control area were 0.39 - 3.92 microg/g fat and 0.46 microg/g (only one sample was detectable), respectively. The TEQ of PCB126 contribute to total TEQ of dioxin-like compounds was similar to other countries and regions. The TEQs of PCBs of the polluted and control area were 59 pg/g fat and 6 pg/g fat, respectively. CONCLUSION: The TEQ of PCBs of polluted area was much higher than other countries and regions, and the dioxin-like compounds intake of infants was much higher than the Tolerable Daily Intake (TDI) for dioxin recommended by WHO. PMID- 17086716 TI - [A cross sectional study of passive smoking of non-smoking women and analysis of influence factors on women passive smoking]. AB - OBJECTIVE: To fund out the state of passive smoking of non-smoking women and search for measures of controlling women passive smoking. METHODS: 3500 non smoking women in Beijing, Shanghai, Chengdu city were interviewed. Analyses were performed by chi2 test Fisher test and ANOVA test. RESULTS: 92.7% passive smoking women exposure to ETS at home, 40.8% at workplace. 38.9% exposed to ETS from birthday, and 42.3% from 18 - 30 age. The average exposure time of passive smoking is (1.17 +/- 1.10) hours per day. The proportion of passive-smoking time over 2 hours at home is higher than work place. In passive-smoking group, the proportion of 30 - 50 age group, secondary education, married, merchant/service, principal of units, and manufacture/transport workers were higher than non smoking group. 97.5% think that passive smoking is harmful to health, and the proportion of thinking passive smoking has severe harm to health in non-passive smoking group is higher than passive-smoking group. 70.0% open windows when someone smokes around her, but only 16.9% ask the smokers do not smoke around her forwardly. Suppose that someone were smoking around yourself, the consciousness of avoiding passive smoking forwardly in non-passive-smoking group is stronger than passive-smoking group. 95.1% believe the content of smoking-harm propagandized by medium. CONCLUSION: The main places of controlling passive smoking are the home and the department, commerce, service, and manufacture/ transport workplace. The rate of passive smoking was influenced by the consciousness of the serious level of harms by passive smoking. Propagandizing the serious harm of passive smoking by medium and strengthening the consciousness of avoiding passive smoking were one of feasible measures to lower the rate of smoking and passive smoking. PMID- 17086717 TI - [Factors influencing snack behaviours of young children in Hefei and evaluation of the effection of interventions]. AB - OBJECTIVE: To evaluate the effect of the parent-to-child and teacher-to-child models used in the Snack Risk Factor Intervention Project for Children (SRFIPC) to change the snack pattern of children. METHODS: Using the community-based intervention trail, nutrition education was implemented based on the parent-to child and teacher-to-child models among 2102 pre-school children aged 4-6 years in 8 day-care centres of Hefei City, Anhui Province. Data on snack consumption of young children was obtained using a snack behaviour questionnaire. RESULTS: (1) The rate of snacking was high in young children of Hefei. The attitude and diet behavior of the parent are the main influence of the young children snack behavior. (2) The snack frequency of children in intervention group decreased as compared with the control group (P < 0.001). However, children who have higher frequent snacking in the intervention group did not have significant change in their snack behaviours compared with children in the control group. (3) The percentage of children who consumed snacks between meals increased, while the percentage of children who consumed snacks ad arbitrium was greatly decreased in the intervention group (both P < 0.001). (4) The percentages of snacks low in nutrition density, such as the biscuit, cake, confectionery, ham sausage, beverages were lower in intervention group than in the control group. CONCLUSION: It is useful to intervention the children's snack behaviours parent-to-child and teacher-to-child models. PMID- 17086718 TI - [Study on the genetic polymorphisms of human glutathione-S-transferase A1 in Hakka population in South China]. AB - OBJECTIVE: Glutathione-S-transferase A1 (GSTA1) is one of the major phase II detoxification enzymes in the cytosol, which genetic polymorphisms distribution is different in different ethnic, national and regional population. Up to now, GSTA1 genetic polymorphisms has been rarely reported in China. The purpose of the study was to investigate the distribution of genetic polymorphisms of human GSTA1 in Hakka population in South China. METHODS: Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) were used to identify the genotypes of GSTA1 gene and the data were analyzed with SPSS10.0 software. RESULTS: The GSTA1 genetic polymorphisms were detected in 480 samples. The frequency of GSTA1 * A/ * A,GSTA1 * A/ * B and GSTA1 * B/ * B were 77.1%, 21.7% and 1.2% respectively. And the GSTA1 genetic polymorphism distribution was in accordance with the Hardy-Weinberg equilibrium rule. There were no difference in the GSTA1 genetic polymorphisms among the different groups of age or gender. Logistic regression analysis showed that there were no association between the GSTA1 genetic polymorphism and family history of hypertension, coronary heart disease, stroke, lung cancer and nasopharyngeal cancer, et al. CONCLUSION: The GSTA1 gene existed polymorphism among Hakka in South China. PMID- 17086719 TI - [Village resident nourishment of Hebei investigates with health condition]. AB - OBJECTIVE: To study the changes of the food and nutrients intake and health trends in rural population of Hebei Province since 1992 with the society economy development rapidly. METHODS: Using the multi-stage cluster random sampling method, investigating the body high, weight, blood pressure, Haemoglobin, blood lipids, blood glucose, of 11627 population from 2700 households, and the food consumption and nutrients intakes of 1080 households, to analyses and assess the dietary pattern and mortality of chronic diseases. RESULTS: The results showed that plant food is main food consumption pattern and changed remarkably since the 1992. Although the meat, egg, milk and beat consumption was more than 1992 their intakes are insufficient. The fat food and the salt intakes increased while CA and VA intakes were very insufficient. Weighs values in the school children aged 7-17 were significantly higher than those in 1992. The prevalence of overweight, obese, high blood pressure, diabetes, abnormal blood sugar were 30.4%, 13.4%, 25.9%, 1.8%, 0.98%, respectively in adults aged over 18. The rate of abnormal blood lipid 25.0%, high TG, TC, verge TC, low HDL-C were 15.3%, 3.2%, 3.5%, 10.8%. The standardized prevalence of anemia 8.7% and 16% for preschool children and younger woman respectively. CONCLUSION: The prevalence of overweight, obese, hypertension and abnormal blood lipids in adult population of rural Hebei Province were higher than the average levels in whole country. Sufficient attention should be paid to nutrition diet, heath and some chronic diseases related to nutrition in rural residents, the dietary guidance and nutrition interference need to strengthen and ensure people health. PMID- 17086720 TI - [Effect of balanced diet on the nutrition status of schoolchildren in the mountain area]. AB - OBJECTIVE: To study the effect of balanced diet on the nutrition status of primary schoolchildren in the mountain area in November 2004. METHODS: All the subjects aged 7-9 years old were divided into intervention group and control group. The balanced diet was provided for the intervention group and the control group was maintained on their usual diet for 30 days in December. The anthropometric and nutritional status of the subjects were checked before and after the study. RESULTS: The growth and nutritional status of the intervention group improved significantly. However the unbalanced diet in the control group affects the normal growth of the children because the usual diet can not meet the nutrient requirement. CONCLUSION: The nutrition intervention can improve the growth and nutritional status of the primary schoolchildren in the mountain area. PMID- 17086721 TI - [Study on the acryl amide intake in the Beijing diet]. AB - OBJECTIVE: A 24-hours recall study was performed to get the acryl amide intake in the Beijing diet. METHODS: 24 separate items bought in the Beijing Market were analyzed and 2460 diet lists (820 participants x 3 days) were calculated. RESULTS: The following mean intakes of acryl amide have been estimated (microg/dm): males 18.6, females 16.6. For the 97.5 percentile of the population the intake is estimated to: males 59.0, females 50.4. As dose calculated per kg bodyweight the mean intake becomes [microg/(kg bw x d)]: males 0.282, females 0.284. For the 97.5 percentile [microg/ (kg bw x d)]: males 0.898, females 1.007. The contribution of the different meals and to the intake was as follows: breakfast 20.48%, lunch 38.99%, dinner 39.42%, snacks 1.17%. CONCLUSION: The survey leads to the conclusion that lunch and dinner are significant sources of acryl amide in a typical Beijing diet even if they have relatively low levels of acryl amide. Chinese traditional breakfast food is also an important source of acryl amide in a typical Beijing. PMID- 17086722 TI - [Study on risk factors of depressive and/or anxiety symptoms in patients with coronary heart disease in general hospitals of urban China]. AB - OBJECTIVE: To explore the risk factors of depressive and/or anxiety symptoms in patients with coronary heart disease (CHD) in the general hospital of main cities in China. METHODS: A hospital-based cross-sectional study was conducted in four main cities in China in 2004. 359 CHD patients were recruited continually during six months from the outpatient and inpatient department of 7 tertiary hospitals. Face-to-face interview was used in data collection together with the self completed HAD scale for depressive and/or anxiety symptom screening. Multinomial Logistic Model was adopted in data analysis. RESULTS: Among 359 CHD patients, 82 (22.8%) obtained a HAD score of 9 and above. Non-ambulatory patients had a statistically higher risk of depressive symptoms (OR = 17.996, 95 % CI:3.872 - 83.636); subjects younger than 65 years old, or having an education year equal or less than 9 year, or having a self-assessment of CHD deterioration had increased risk of anxiety symptoms with the ORs at 3.151 (95% CI: 1.151-8.629), 3.154(95% CI: 1.094-9.092) and 4.229 (95% CI: 1.396-12.809) respectively. Hospitalized (OR = 4.887, 95% CI: 1.711-13.960) and non-ambulatory (OR = 6.583, 95% CI: 2.776 15.612) CHD patients were more liable to suffer both depressive and anxiety symptoms. CONCLUSION: It was noticeable to health care providers both in somatic disease care and mental health care that non-ambulatory status, younger than 65 years old, lower education level, felt deterioration of CHD and hospitalization were possible risk factors for depressive and/or anxiety symptoms in patients with CHD in general hospitals in urban China. PMID- 17086723 TI - [A method for determination of volatile organic compounds in drinking water by purge and trap in combination with gas chromatograph/mass spectrometry]. AB - OBJECTIVE: To establish a method of purge and trap in combination with gas chromatograph/mass spectrometry(P and T-GC/MS) for the rapid determination of minimum multi-mixture of volatile organic compounds (VOCs) in drinking water simultaneously. METHODS: The experimental conditions of P and T, such as desorption temperature, desorption time, the type of the trap and the analytical conditions of GC were optimized. 54 VOCs in drinking water were determined by P and T-GC/MS. RESULTS: The method enjoyed a wide linear range and good precision, the method detection limit(MDL) was 0.01 - 0.5 microg/L, the average recoveries of the method was 90% - 110% and the relative standard deviation (RSD) was 1.24% 7.79%. CONCLUSION: The results showed that the method was accurate and high sensitivity and might be good for application and suitable for trace analysis of VOCs in drinking water. PMID- 17086724 TI - [Study on the predictive model for Yersinia enterocolitica growth at different temperatures]. AB - OBJECTIVE: A predictive model for Yersinia enterocolitica growth in MTSB at different temperatures was studied. According to this model, if food has been contaminated by a few Yersinia enterocolitica, the growth rate, generation time, lag time and total bacterial number at any time and any effective temperature can be predicted. Under the same condition, the parameters for Yersinia enterocolitica serotype O:3 and Yersinia enterocolitica O:9 were compared, respectively. RESULTS: 1. The minimum growth temperature for Yersinia enterocolitica serotype O:3 and O:9 were--17.8 degrees C and--10.3 degrees C, respectively. 2. In the range of 4 degrees C and 30 degrees C, the growth rate of Yersinia enterocolitica serotype O:3 or O:9 increase with temperature going up. 3. In the range of 4 degrees C and 15.7 degrees C, the growth rate of Yersinia enterocolitica serotype O:3 is higher than that of Yersinia enterocolitica serotype O:9. In the range of 15.7 degrees C and 30 degrees C, the growth rate of Yersinia enterocolitica serotype O:9 is higher than that of Yersinia enterocolitica serotype O:3. 4. We get a predictive model for Yersinia enterocolitica in MTSB: Yersinia enterocolitica serotype O:3, square root of M = 0.0134 (T-255.1866). Yersinia Enterocolitica serotype O:9, square root of M = 0.0173 (T-262.7457). PMID- 17086725 TI - [Estrogenic activities of parabens]. AB - OBJECTIVE: Studies indicated that parabens, used as anti-microbial agents in food, cosmetics, and pharmaceuticals, produced a positive uterotrophic response in vivo. They also damaged the late stages of spermatogenesis, altered proportion of pups born alive, and body weight of offspring. They reduced the number of sperm in the epididymis, and the sperm motile activity in male offspring. Parabens could compete with [3H] 17beta-estradiol for binding to the estrogen receptor. The proliferation of two estrogen-dependent cell lines MCF-7 and ZR-75 1 could be increased by parabens. They also increased expression of both transfected and endogenous estrogen-regulated genes in MCF-7 cells. The studies demonstrated parabens were weakly estrogenic. PMID- 17086726 TI - [Effects of formaldehyde on the activity of superoxide dismutases and glutathione peroxidase and the concentration of malondialdehyde]. AB - This review focus on the effects of formaldehyde on the activity of superoxide dismutases (SOD), glutathione peroxidase (GSH-Px) and the concentration of malondialdehyde (MDA) in recent years. It is showed formaldehyde having effect on lipid peroxidation by many studies of animal experiment and occupational group, it can decrease the activity of SOD and GSH-Px, increase the concentration of MDA, there are dose-response relationship between those lipid peroxidation index and the concentration of formaldehyde too. In future study, the animal experiment and the people's late effect of low dose formaldehyde exposure should be pay more attention. PMID- 17086727 TI - [Adverse health effects of cadmium and related factors]. AB - Cd is a very toxic chemical to animal and human beings. Cd contamination has been ubiquitous because of industry and the human activities. Cd not only affects adversely the growth, yield and quality of crops but also effects on the health of animal and human being. The damage of Cd on bone, kidney, liver, immune and procreation systems as well possible factors that influence Cd absorption, accumulation and damage in the body are summarized in this paper. PMID- 17086728 TI - [Studies on the nephrotoxicity of chromium compounds]. AB - The toxicity of chromium in different valent are quite different. Cr (III) is a necessary trace element of human body, but excessive intake of Cr (III) may bring about health lesion. Cr (VI) is generally considered 1,000 times more toxic than Cr (III). Renal damge may happen when people contact or intake excessive chromium compounds. The renal function and the concentration of enzyme and protein may be changed. Kidney necrosis can happen in heavy cases. The metaboly of chromium compounds and its nephrotoxicity were reviewed in this paper. PMID- 17086729 TI - [Toxic effects and the mechanism of mercury on health]. AB - Mercury is an unnecessary element for human body, and it can exist in several physical and chemical forms in the environment. All forms of mercury have toxic effects in a number of organs, especially in the neural system and kidney. Now, with its extensive use in the industry and life, people pay more attention on the health effects which are caused by mercury. In order to know more about it and to make further study in the future, this review will make a summarize on the toxicity and its mechanism of mercury. PMID- 17086730 TI - [Assessment methods and evolution on vitamin A status]. AB - Vitamin A is one of nutrients which are indispensable to all people, especially children and pregnant women. In the world, severe vitamin A deficiency (VAD) has been controlled efficiently. But to many countries, especially those developing, subclinical or marginal vitamin A deficiency is still a public health problem. It is very important and necessary to assess the status of vitamin A of the population who are in risk of VAD before drawing and carrying out the intervention strategy. Currently, clinical, histological, functional and biochemical methods can be used to evaluate the level of vitamin A. PMID- 17086731 TI - [Simultaneous determination methods of biogenic amines in food]. AB - Biogenic amines are organic bases with low molecular weight. They can be expected in all foods that contain proteins or free amino acids. Consumption of food containing high amounts of biogenic amines may cause toxicological effects. The most notorious foodborne intoxications caused by biogenic amines are related to histamine. The article reviewed methods of determination of biogenic amines in foods in recently years and emphasized on the characteristics of HPLC, CE, biosensor and TLC and their applications. PMID- 17086732 TI - Quantitative assessment of myocardial perfusion defects with real-time three dimensional myocardial contrast echocardiography. AB - OBJECTIVE: To evaluate the feasibility and accuracy of measurement of myocardial perfusion defects with intravenous contrast-enhanced real-time three-dimensional echocardiography (CE-RT3DE). METHODS: RT3DE was performed in 21 open-chest mongrel dogs undergoing acute ligation of the left anterior descending artery (LAD, n = 14) or distal branch of the left circumflex artery (LCX, n = 7). A perfluorocarbon microbubble contrast agent was injected intravenously to assess the resulting myocardial perfusion defects with Philips Sonos-7500 ultrasound system. Evans blue dye was injected into the occluded coronary artery for subsequent anatomic identification of underperfused myocardium. In vitro anatomic measurement of myocardial mass after removal of the animal's heart was regarded as the control. Blinded off-line calculation of left ventricular mass and perfusion defect mass from RT3DE images were performed using an interactive aided manual tracing technique. RESULTS: Total left ventricular (LV) myocardial mass ranged from 38.9 to 78.5 (mean +/- SD: 60.0 +/- 10.1) g. The mass of perfusion defect ranged from 0 to 21.4 (mean +/- SD: 12.0 +/- 5.0) g or 0 to 27% of total LV mass (mean +/- SD: 19% +/- 6%). The RT3DE estimation of total LV mass (mean +/ SD: 59.8 +/- 9.9 g) strongly correlated with the anatomic measurement (r = 0.98; y = 2.01 + 0.96x). The CE-RT3DE calculation of the mass of underperfused myocardium (mean +/- SD: 12. 3 +/- 5.3 g) also strongly correlated with the anatomic measurement (r = 0. 96; y = - 0.10 + 1.04x) and when expressed as percentage of total LV mass (r = 0.95; y = -0.20 + 1.04x). CONCLUSIONS: RT3DE with myocardial contrast opacification could accurately estimate underperfused myocardial mass in dogs of acute coronary occlusion and would play an important role in quantitative assessment of myocardial perfusion defects in patients with coronary artery disease. PMID- 17086733 TI - Intracranial arterial occlusive lesion in patients with Graves' disease. AB - OBJECTIVE: To investigate the distribution and clinical manifestations of intracranial arterial occlusive lesions (IAOLs), and their correlation with thyroid function. METHODS: We enrolled 7 patients who had Graves' disease (GD) with IAOLs screened and evidenced by transcranial Doppler, then further confirmed with digital substract angiography in 2 patients and magnetic resonance angiography in 5 patients. Brain magnetic resonance imaging (MRI) was performed in all 7 patients. Three patients were followed up. RESULTS: Among 7 patients, 1 was male and 6 were females. The mean age was 32.0 +/- 5.5 (range from 11 to 49) years old. Six of them had symptoms of GD but one was asymptomatic with abnormality of T3, T4, and thyroid stimulating hormone. The lesions of intracranial arteries were symmetrical bilaterally in the internal carotid artery system in 6 patients, as well as asymmetrical in 1 patient Terminal internal carotid artery (TICA) were involved in all 7 patients. Middle cerebral artery (MCA) were involved in 3, anterior cerebral artery in 2, and basilar artery in 1 patient. Net-like collateral vessels and mimic moyamoya disease were observed in the vicinity of the occlusive arteries in 2 patients. All patients presented symptoms of ischemic stroke including transient ischemic attack and/or infarction while IAOLs were found. Three patients had obvious involuntary movements. Brain MRI revealed infarctions located in the cortex, basal ganglion, or hemiovular center in 5 patients. The remaining 2 patients had normal brain MRI. The neurological symptoms were improved concomitant with relief of the thyroid function in 2 patients, while IAOLs were aggravated with deterioration of the thyroid function in 1 patient. CONCLUSION: IAOLs in patients with GD mainly involve intracranial arteries, especially the TICA and MCA, which is similar to moyamoya disease. The neurological symptoms and severity of involved arteries may relieve while the hyperthyroidism is gradually under control. PMID- 17086734 TI - Low dose pirfenidone suppresses transforming growth factor beta-1 and tissue inhibitor of metalloproteinase-1, and protects rats from lung fibrosis induced by bleomycina. AB - OBJECTIVE: To investigate the optimal dosage of pirfenidone for the treatment of pulmonary fibrosis induced by bleomycin in Wistar rats, and the alteration of expressions of transforming growth factor beta-1 (TGF-beta 1), tissue inhibitor of metalloproteinase-1 (TIMP-1), and matrix metalloproteinase-13 (MMP-13) in lung tissue. METHODS: Male Wistar rats were endotracheally instilled with bleomycin or normal saline. Pirfenidone (25-800 mg x kg(-1) x d(-1)), dexamethasone (3 mg/kg), or 1% carboxymethylcellulose sodium were given daily by feed 2 days before instillation of bleomycin. Groups T7 and T14 were fed pirfenidone 50 mg x kg(-1) x d(-1) at 7 days or 14 days after bleomycin instillation. Lungs were harvested at 28 days after bleomycin instillation. Patholological changes in lung tissues were evaluated with HE staining. Lung collagen was stained by sirius red and measured by content of hydroxyproline. Expression of proteins of TGF-beta 1, TIMP 1, and MMP-13 were detected by Western blotting. RESULTS: At doses of 25, 50, and 100 mg x kg(-1) x d(-1), pirfenidone had significant anti-fibrotic effects for bleomycin-induced rat pulmonary fibrosis, and these effects were most significantly attenuated at the dosage of 50 mg x kg(-1) x d(-1) (HE: P < 0.01, P < 0.01, and P = 0.064; sirius red: P < 0.05, P < 0.01, and P < 0.05; hydroxyproline: P = 0.595, P < 0.01, and P = 0.976). Pirfenidone at a dosage of 50 mg x kg(-1) x d(-1) inhibited protein expression of TGF-beta1 and TIMP-1 in lung tissue in the early phase (0.79 and 0.75 times of control group), but had no effect on expression of MMP-13. CONCLUSION: Low dose pirfenidone, especially at dosage of 50 mg x kg(-1) x d(-1), has significant anti-fibrotic effects on bleomycin-induced rat pulmonary fibrosis. Pirfenidone partially inhibits the enhancement of the expression of TGF-beta 1 and TIMP-1 in lung tissue. PMID- 17086735 TI - CXC chemokine receptor 3 modulates bleomycin-induced pulmonary injury via involving inflammatory process. AB - OBJECTIVE: To investigate the role of CXC chemokine receptor 3 (CXCR3) in bleomycin-induced lung injury by using CXCR3 gene deficient mice. METHODS: Sex-, age-, and weight-matched C57BL/6 CXCR3 gene knockout mice and C57BL/6 wide type mice were challenged by injection of bleomycin via trachea. Lung tissue was stained with HE method. Airway resistance was measured. Bronchoalveolar lavage (BAL) was performed using phosphate buffered saline twice, cell number and differentials were counted by Diff-Quick staining. Interleukin (IL)4, IL-5, IL 12p40, and interfon-y in BAL fluid and lung homogenate were measured by enzyme linked immunosorbent assay. Unpaired t test was explored to compare the difference between two groups. RESULTS: On day 7 after bleomycin injection via trachea, CXCR3 knockout mice were protected from bleomycin-induced lung injury as evidenced by fewer accumulation of inflammatory cells in the airway and lung interstitium compared with their wild type littermates (P < 0.05). Airway resistance was also lower in CXCR3 knockout mice compared with wild type mice (P < 0.01). Significantly lower level of inflammatory cytokines release, including the altered production of IL-4 and IL-5 both in BAL fluid and lung tissue was seen in CXCR3 knockout mice than in wild type mice (both P<0.05). CONCLUSION: CXCR3 signaling promotes inflammatory cells recruiting and initiates inflammatory cytokines cascade following endotracheal bleomycin administration, indicating that CXCR3 might be a therapeutic target for pulmonary injury. PMID- 17086736 TI - Mitomycin C "straight scleral tunnel incision"--trabeculectomy with a releasable suture. AB - OBJECTIVE: To evaluate the efficacy of "straight scleral tunnel incision" trabeculectomy with a releasable suture supplemented with mitomycin C (MMC) on reducing intraocular pressure (IOP), complications, and corneal astigmatism in patients with primary angle-closure glaucoma (PACG). METHODS: Totally 217 acute or chronic PACG patients with occludable angle above 180 degrees and IOP above 21 mm Hg were divided into 3 groups. Patients in group A (98 cases, 128 eyes), B (71 cases, 95 eyes), and C (48 cases, 60 eyes) were treated with "straight scleral tunnel incision"-trabeculectomy with a releasable suture supplemented with MMC, releasable suture trabeculectomy with MMC, and trabeculectomy with MMC, respectively. IOP, complications, and surgically induced astigmatism (SIA) were evaluated preoperatively and up to 12 months postoperatively. RESULTS: IOP of 2 weeks after treatment was significantly lower than preoperative IOP in all the 3 groups (all P <0. 001). Success rates (IOP < or = 20 mm Hg) in group A, B, and C were 87.91%, 89.23%, and 83.72% respectively at 12 months after treatment (P = 0.256). The incidence of shallow anterior chamber and hypotony had no significant difference between group A and B, but both of them were lower than that in group C (P < 0.05). There were no significant differences in preoperative corneal astigmatism among the 3 groups. The corneal astigmatism after 2 weeks in group A (1.71 +/- 1.47D) was higher than that before operation (1.28 +/- 1.05D, P = 0.126). With 12 months gone, the astigmatism almost returned to preoperative levels. The corneal astigmatisms after 2 weeks in group B and C (1.99 +/- 1.20D and 2. 22 +/- 1.39D) were significantly higher than those before operation (1.20 +/- 0.85D and 1.18 +/- 0.93D, P = 0.002, P = 0.001), respectively. With 12 months gone, the mean astigmatisms in group B and C (1.87 +/- 0.91D and 1.90 +/- 1.16D) were still significantly higher than those before operation (P = 0.001, P = 0.003). The highest astigmatic polar values in group A, B, and C (1.00D, 1. 89D, and 1. 77D) occurred after 2 weeks, 1 month, and 1 month postoperation, respectively, which were significantly higher than those before operation (0.19 +/- 1.32D, 0.12 +/- 1.22D, and 0.17 +/- 1.25D, P < 0.01), respectively. With 12 months gone, they were 0. 03D, -0. 18D, and -0. 13D higher than those before operation, respectively. The rates of function bleb and thin-wall bleb were 71.43% and 26.37% in group A, 75.38% and 29.23% in group B, 72.09% and 25.58% in group C, respectively at 12 months after treatment There were no significant differences among the 3 groups. CONCLUSION: "Straight scleral tunnel incision" trabeculectomy with a releasable suture supplemented with MMC can reduce complications and get satisfactory results in reducing IOP and SIA. PMID- 17086737 TI - Evaluation of oral midazolam as conscious sedation for pediatric patients in oral restoration. AB - OBJECTIVE: To evaluate the effect of midazolam alone on sedation in young children for dental restorative care. METHODS: Forty children, aged 5 to 10 years with a mean age of 7.3 years, participated in this study. Twenty-one patients were assigned to intervention group received 0. 5 mg/kg of oral midazolam 20 minutes prior to the beginning of dental treatment, and 19 patients in control group received placebo liquid 20 minutes before treatment All patients received painless local anesthetic injection and were restrained with children's board and bands. Blood pressure (BP), heart rate (HR), oxygen saturation, treatment compliance scores of the Ramsay scale, the Briekopf and Buttner scale, Frankl scale, and the Houpt scale were recorded. Each procedure was taped and all the data were evaluated every 5 minutes by an anesthetist or experienced dentist who was unaware of the drug given to the child. RESULTS: HR in intervention group (82.5 +/- 5.1 bpm) was much lower than that in control group (95.2 +/- 8.9 bpm; F = 31.20, P < 0.001). Intervention group had a significantly lower systolic BP level (94.8 +/- 5.6 mm Hg) than control group (98.5 +/- 5.5 mm Hg; F = 4.34, P = 0.04), but the diastolic BP (63.0 +/- 3.5 mm Hg) was not significantly lower than control group (65.5 +/- 4.8 mm Hg; F = 3.31, P = 0.07). Children in intervention group showed more compliance. The patients' scores of the Ramsay scale, Briekopf and Buttner scale, Frankl scale, and Houpt scale in intervention group (1.37 +/- 0.96, 1.37 +/- 0.83, 1.32 +/- 0.67, and 2.32 +/- 1.49, respectively) were significantly lower than those in control group (3.71 +/- 1.23, 2.71 +/- 0.96, 2.71 +/- 0.90, and 4.71 +/- 1.19; F = 44.66, 22.36, 30.39, and 31.88, respectively, all P < 0.001). CONCLUSIONS: Oral midazolam alone is safe and produces effective sedation for the dental treatment of young children. Oral midazolam application should be generally preferred because it is more easily accepted by pediatric patients. PMID- 17086738 TI - Correlation between fibrinogen level and cerebral infarction. AB - OBJECTIVE: To investigate the correlation between plasma fibrinogen level and cerebral infarction (CI) as well as the difference of fibrinogen among subtypes of CI. METHODS: A case-controlled study was conducted with 131 cases of CI and 148 controls. Plasma fibrinogen levels were detected by the Clauss method. RESULTS: High fibrinogen level (3.09 +/- 0.94 g/L) was correlated with CI (OR = 2.47, 95% CI: 1.51-4.04, P < 0.005) at the onset stage of the disease. Persistent high fibrinogen level (3.14 +/- 0.81 g/L) at 6-month after stroke onset was detected and correlated with CI (OR = 4.34, 95% CI: 1.80-10.51, P = 0.001). Higher fibrinogen level was correlated with total anterior circulation infarction (TACI), partial anterior circulation infarction (PACI), and posterior circulation infarction (POCI) (OR = 4.008, P < 0.001). Higher fibrinogen level was correlated with extracranial atherosclerosis (OR = 3.220, P < 0.05, but not with intracranial atherosclerosis. CONCLUSION: Fibrinogen level may be a risk factor of CI and probably correlates with subtypes of CI and distributions of atherosclerosis. PMID- 17086739 TI - Leukocytosis and retinoic acid syndrome in patients with acute promyelocytic leukemia treated with arsenic trioxide. AB - OBJECTIVE: To study the incidence of leukocytosis and retinoic acid (RA) syndrome in newly diagnosed and relapsed acute promyelocytic leukemia (APL) patients treated with arsenic trioxide (ATO). METHODS: Thirty patients with newly diagnosed or relapsed APL received ATO for remission induction at the dose of 10 mg/d. RA syndrome was defined when patient was with one or more of the following signs or symptoms: fever, dyspnea, serous cavity effusion, muscular pain, pulmonary infiltration, weight gain, or pulmonary infiltration on chest X-ray. RESULTS: Twenty-three (77% ) patients achieved complete remission, mean time to remission was 37.1 days. Leukocytosis was observed in 14 (47%) patients, mean time to leukocytosis was 12.7 days, median baseline leukocyte count for patients with leukocytosis was 3.1 x 10(9)/L, which was higher than that for patients who did not develop leukocytosis (2.6 x 10(9)/L, z = -2.635, P = 0.008). No other cytotoxic therapy was administered, and the leukocytosis resolved in all cases. The RA syndrome was observed in 9 (30%) patients, mean time to diagnose of RA syndrome was 13.9 days, median baseline leukocyte count for patients with RA syndrome was 3.6 x 10(9)/L, which was higher than that for patients who did not develop RA syndrome (2.6 x 10(9)/L, z = -1.909, P = 0.046). No patient died of RA syndrome. CONCLUSION: Leukocytosis and RA syndrome are associated with ATO and baseline leukocyte count respectively, and there is distinct link between leukocytosis and RA syndrome. PMID- 17086740 TI - Serum concentrations of hyaluronic acid, procollagen type III NH2-terminal peptide, and laminin in patients with chronic congestive heart failure. AB - OBJECTIVE: To explore the role of serum fibrotic indices including hyaluronic acid (HA), procollagen type III NH2 -terminal peptide (PCIIIP), and laminin (LN) in assessing the severity of myocardial fibrosis in chronic congestive heart failure (CHF). METHODS: Serum levels of HA, PCIIIP, and LN in 39 patients with CHF [14 with New York Heart Association (NYHA) functional class II, 21 with class III, 4 with class IV] and in 46 patients with NYHA functional class I were assessed by radioimmunoassay. RESULTS: The serum concentrations of HA, PCIIIP, and LN were 359.75 +/- 84.59 microg/L, 77.88 +/- 24.67 microg/L, 86.73 +/- 23.90 microg/L in CHF group, and 211.60 +/- 54. 80 microg/L, 64.82 +/- 23.99 microg/L, 82.26 +/- 23.98 microg/L in NYHA functional class I group, respectively. The HA level was significantly higher in CHF patients as compared with NYHA functional class I group (P < 0.05). However, no difference was found in the levels of PCIIIP and LN between CHF group and NYHA functional class I group. The serum HA concentration was negatively correlated with left ventricular ejection fraction (r = - 0.71, P < 0.05). CONCLUSION: Serum HA level may act as an indicator for myocardial fibrosis. PMID- 17086741 TI - Influence of quercetin and x-ray on collagen synthesis of cultured human keloid derived fibroblasts. AB - OBJECTIVE: To investigate the effects of quercetin and X-ray on collagen synthesis of cultured human keloid-derived fibroblast and the mechanism. METHODS: Collagen synthesis of cultured human keloid and normal fibroblasts were detected by hydroxyproline colorimetric analysis. Immunocytochemical staining was used to investigate collagen I and III expression. mRNA expression of collagen I and III, and transforming growth factor (TGF)-beta 1 were assayed by reverse transcription polymerase chain reaction (RT-PCR) and real-time PCR. RESULTS: Quercetin inhibited the collagen synthesis of both keloid and normal fibroblasts in a dose dependent manner. Immunocytochemical staining indicated that collagen I and III were down-regulated by quercetin and X-ray (P < 0.05), particularly collagen I (P < 0.05). mRNA expression of both collagen I and III in quercetin groups significantly decreased compared with that in control group (P < 0.05), especially in the group treated with both quercetin and X-ray (P < 0.01). mRNA level of TGF-beta 1 gene was down-regulated by quercertin (P < 0.05). CONCLUSIONS: Quercetin will probably be one of the new medicines which could effectively treat keloid. Quercetin combined with X-ray could reduce the dose of radiation. PMID- 17086743 TI - It is gastroesophageal reflux disease, not asthma: a case report. PMID- 17086742 TI - Experimental study on plasticity of proliferated neural stem cells in adult rats after cerebral infarction. AB - OBJECTIVE: To investigate whether there is endogenous neural stem cell proliferation and whether these proliferated neural stem cells represent neural plasticity in the adult rats after cerebral infarction. METHODS: Cerebral infarction models of rats were established and the dynamic expression of bromodeoxyuridine (BrdU), BrdU/polysialylated neural cell adhesion molecule (PSA NCAM) were determined by immunohistochemistry and immunofluorescence staining. BrdU was used to mark dividing neural stem cells. PSA-NCAM was used to mark the plasticity of neural stem cells. RESULTS: Compared with controls, the number of BrdU-positive cells in the subventricular zone (SVZ) and hippocampus increased significantly at 1st day after cerebral infarction (P < 0.05), reached maximum at 7th day, decreased markedly at 14th day, but it was still elevated compared with that of the controls (P < 0.05). The number of BrdU-labeled with PSA-NCAM positive cells increased significantly at 7th day (P < 0.05), reached maximum at 14th day, markedly decreased at 28th day, but it was still elevated compared with that of the controls (P < 0.05). It was equal to 60% of the number of BrdU positive cells in the same period. CONCLUSION: Cerebral infarction may stimulate the proliferation of endogenous neural stem cells in situ and most proliferated neural stem cells represent neural plasticity. PMID- 17086744 TI - Successful extracorporeal membrane oxygenation support in a patient with fulminant myocarditis. PMID- 17086745 TI - An unusual bilateral vagal paragangliomas: one case study. PMID- 17086746 TI - Diagnosis and treatment of mediastinal enterogenous cysts in children. AB - OBJECTIVE: To investigate the diagnosis and therapy of mediastinal enterogenous cysts in children. METHODS: Clinical data of 17 cases with mediastinal enterogenous cysts within 19 years in our hospital were retrospectively analyzed. RESULTS: One case was intramural esophageal cyst and 16 cases were enteric cysts, two among which were complicated with abdominal enteric duplications. Most cases presented with symptoms of respiratory distress. Twelve cases were complicated with vertebral anomalies. Ultrasound of 12 cases and magnetic resonance imaging of 4 cases were helpful in confirming the cystic nature of these lesions. Eight cases had technetium-99m pertechnetate scintigraphy of posterior mediastinum. CONCLUSIONS: Most patients present with symptoms of respiratory distress, complicated with vertebral anomalies. Ultrasonography and magnetic resonance imaging may be helpful in confirming the cystic nature of these lesions. Technetium-99m pertechnetate scintigraphy is the most effective method for differentiation of the disease from other mediastinal cysts. PMID- 17086747 TI - Docetaxel influences autocrine of transforming growth factors and induces apoptosis in human ovarian cancer cell line AO. PMID- 17086748 TI - Needle cell elongation and maturation timing derived from pine needle cellulose delta18O. AB - Estimates of the timing of Pinus arizonica Engelm. needle development in 1998 and 1999 were derived from the leaf-cellulose delta18O of weekly growth increments. Significant correlations were noted between time series of local humidity and leaf-cellulose delta18O for needles growing near Tucson, Arizona. Correlations with temperature were also significant, but much lower, suggesting these variations in cellulose delta18O were determined mostly by changes in humidity. The timing of all significant correlations lags the timing of the appearance of the new needle growth, and is interpreted as indicating 16-23 d were required for cell enlargement in 1998 and 13-17 d in 1999. Similarly, properties of the environmental time series, when significantly correlated, are interpreted as indicating the duration of cellulose deposition (7-27 d in 1998, 13-21 d in 1999). Variations in stable-isotope back diffusion (the Peclet effect) and the synthesis of cellulose using stored photosynthate are discussed as explanations for departures from a Craig and Gordon-type model of leaf water delta18O. The Peclet effect, use of stored photosynthate, and variations in the growing-season source-water delta18O, probably confound the development of a high-resolution paleohumidity proxy from subfossil needle cellulose delta18O in this region. PMID- 17086749 TI - Effects of cold-girdling on flows in the transport phloem in Ricinus communis: is mass flow inhibited? AB - The effects of cold girdling of the transport phloem at the hypocotyl of Ricinus communis on solute and water transport were investigated. Effects on the chemical composition of saps of phloem and xylem as well as of stem tissue were studied by conventional techniques and the water flow in the phloem was investigated by NMR imaging. Cold girdling reduced the concentration of sucrose but not that of inorganic solutes or amino acids in phloem saps. The possibility that cold treatment inhibited the retrieval of sucrose into the phloem, following leaching from the sieve tubes along a chemical gradient is discussed. Leaching of other solutes did not occur, as a result of missing promoting gradients in stem tissue. Following 3 d of cold girdling, sugar concentration increased and starch was synthesized and accumulated in stem tissue above the cold girdling region and along the cold-treated phloem pathway due to leaching of sugars from the phloem. Only in the very first period of cold girdling (<15-30 min) was mass flow inhibited, but recovered in the rest of cold treatment period to values similar to the control period before and the recovery period after the cold treatment. It is concluded that cold treatment affected phloem transport through two independent and reversible processes: (1) a permanent leaching of sucrose from the phloem stem without normal retrieval during cold treatment, and (2) a short term inhibition of mass flow at the beginning of cold treatment, possibly involving P proteins. Possible further mechanisms for reversible inhibition of water flow are discussed. PMID- 17086750 TI - Diurnal and seasonal variation in root xylem embolism in neotropical savanna woody species: impact on stomatal control of plant water status. AB - Vulnerability to water-stress-induced embolism and variation in the degree of native embolism were measured in lateral roots of four co-occurring neotropical savanna tree species. Root embolism varied diurnally and seasonally. Late in the dry season, loss of root xylem conductivity reached 80% in the afternoon when root water potential (psi root) was about -2.6 MPa, and recovered to 25-40% loss of conductivity in the morning when psi root was about -1.0 MPa. Daily variation in psi root decreased, and root xylem vulnerability and capacitance increased with rooting depth. However, all species experienced seasonal minimum psi root close to complete hydraulic failure independent of their rooting depth or resistance to embolism. Predawn psi root was lower than psi soil when psi soil was relatively high (> -0.7 MPa) but became less negative than psi soil, later in the dry season, consistent with a transition from a disequilibrium between plant and soil psi induced by nocturnal transpiration to one induced by hydraulic redistribution of water from deeper soil layers. Shallow longitudinal root incisions external to the xylem prevented reversal of embolism overnight, suggesting that root mechanical integrity was necessary for recovery, consistent with the hypothesis that if embolism is a function of tension, refilling may be a function of internal pressure imbalances. All species shared a common relationship in which maximum daily stomatal conductance declined linearly with increasing afternoon loss of root conductivity over the course of the dry season. Daily embolism and refilling in roots is a common occurrence and thus may be an inherent component of a hydraulic signaling mechanism enabling stomata to maintain the integrity of the hydraulic pipeline in long-lived structures such as stems. PMID- 17086751 TI - JrSUT1, a putative xylem sucrose transporter, could mediate sucrose influx into xylem parenchyma cells and be up-regulated by freeze-thaw cycles over the autumn winter period in walnut tree (Juglans regia L.). AB - Sucrose has been reported to play multiple roles in the winter biology of temperate woody species. However, no report on the molecular basis of sucrose transport in xylem tissue has yet been made. In the walnut tree, it is demonstrated that during the autumn-winter period, active absorption of sucrose from xylem vessels to parenchyma cells (sucrose influx) is much higher when samplings were taken shortly after a period of freezing temperatures. Here, the question of whether this increased sucrose influx mirrors a regulation of sucrose transporters in xylem tissue was tested. A putative sucrose transporter cDNA (JrSUT1: Juglans regia sucrose transporter 1) was isolated. Over the autumn winter period, JrSUT1 transcripts and respective proteins were present in xylem parenchyma cells and highly detected when samplings were performed shortly after a freeze-thaw cycle. This up-regulation of JrSUT1 level was confirmed in controlled conditions and was not obtained in bark. Immunolocalization studies showed that JrSUT1 and plasma membrane H+ -ATPase (JrAHA) were colocalized to vessel-associated cells (VACs), which control solute exchanges between parenchyma cells and xylem vessels. We propose that JrSUT1 could be involved in the retrieval of sucrose from xylem vessel. All these data are discussed with respect to the winter biology of the walnut tree. PMID- 17086752 TI - Decreased SBPase activity alters growth and development in transgenic tobacco plants. AB - The effects of reduced SBPase activity on growth and development were examined in a set of transgenic tobacco plants produced using an antisense construct driven by the ribulose bisphosphate carboxylase, small subunit promoter. Photosynthetic carbon assimilation rates and carbohydrate levels in source leaves were decreased in the antisense plants. Growth rate and total shoot biomass were reduced in the SBPase antisense plants, even in plants where SBPase activity was reduced by only 25%. Floral biomass also decreased in response to reductions in SBPase activity and the onset of flowering was delayed by 5-10 d. This is the first demonstration of a link between reproductive biomass and reductions in Calvin cycle enzyme activity using antisense plants. Furthermore, unexpected changes in the growth and development of the antisense plants were evident. Small reductions in SBPase activity (above 50% wild type) resulted in shorter plants with only a small decrease in stem biomass and specific leaf area. In contrast, plants with larger reductions in SBPase activity had an increase in specific leaf area and attained heights similar to that of the wild-type plants but with a much reduced stem biomass, largely due to a decrease in xylem tissue. This bi-modal response of growth to reductions in SBPase activity has similarities to changes in leaf and stem anatomy and morphology that accompany light acclimation. PMID- 17086753 TI - Nitric oxide (NO) as an intermediate in the cryptogein-induced hypersensitive response--a critical re-evaluation. AB - A hypersensitive response (HR) was induced in tobacco leaves and cell suspensions by the fungal elicitor cryptogein, and NO production was followed by chemiluminescence and occasionally by diaminofluorescein (DAF)-fluorescence. Results from both methods were at least partly consistent, but kinetics was different. NO emission was not induced by cryptogein in leaves, whereas in cell suspensions some weak NO emission was observed, which was nitrate reductase (NR) dependent, but not required for cell death. Nitric oxide synthase (NOS) inhibitors did not prevent cell death, but PR-1 expression was weakened. In conclusion, neither NR nor NOS appear obligatory for the cryptogein-induced HR. However, a role for NO was still suggested by the fact that the NO scavenger cPTIO prevented the HR. Unexpectedly, cPTI, the reaction product of cPTIO and NO, also impaired the HR but without scavenging NO. Thus, prevention of the HR by cPTIO is not necessarily indicative for a role of NO. Further, even a 100-fold NO overproduction (over wild type) by a nitrite reductase-deficient mutant did not interfere with the cryptogein-induced HR. Accordingly, the role of NO in the HR should be reconsidered. PMID- 17086754 TI - Water flow through junctions in Douglas-fir roots. AB - Roots are important conduits for the redistribution of water within the rooting zone. Root systems are often highly branched, and water flow between regions undoubtedly involves passage through junctions between individual roots. This study considered junctions in the roots of Douglas-fir with regard to the resistances encountered by water flow through the xylem. Flow into the root branch distally along the main root encountered much greater resistance than flow into the branch and proximally along the main root (toward the plant stem). When the main root proximal to the junction was gradually shortened, the resistance to flow in the branch root and distally along the main root increased dramatically. Thus, flow in this manner appears to depend on lateral flow within the root over many centimetres proximal to the junction and not just within the direct connection at the junction. These results suggest that the hydraulic nature of junctions is an important aspect of hydraulic redistribution of water within the soil utilizing flow through roots. PMID- 17086755 TI - Contributions of evaporation, isotopic non-steady state transpiration and atmospheric mixing on the delta18O of water vapour in Pacific Northwest coniferous forests. AB - Changes in the 2H and 18O of atmospheric water vapour provide information for integrating aspects of gas exchange within forest canopies. In this study, we show that diurnal fluctuations in the oxygen isotope ratio (delta 18O) as high as 4% per hundred were observed for water vapour (delta (18)Ovp) above and within an old-growth coniferous forest in the Pacific Northwest region of the United States. Values of delta 18Ovp decreased in the morning, reached a minimum at midday, and recovered to early-morning values in the late afternoon, creating a nearly symmetrical diurnal pattern for two consecutive summer days. A mass balance budget was derived and assessed for the 18O of canopy water vapour over a 2-d period by considering the 18O-isoflux of canopy transpiration, soil evaporation and the air entering the canopy column. The budget was used to address two questions: (1) do delta 18O values of canopy water vapour reflect the biospheric influence, or are such signals swamped by atmospheric mixing? and (2) what mechanisms drive temporal variations of delta 18Ovp? Model calculations show that the entry of air into the canopy column resulted in an isotopically depleted 18O-isoflux in the morning of day 1, causing values of delta 18Ovp, to decrease. An isotopically enriched 18O-isoflux resulting from transpiration then offset this decreased delta 18Ovp later during the day. Contributions of 18O-isoflux from soil evaporation were relatively small on day 1 but were more significant on day 2, despite the small H2(16)O fluxes. From measurements of leaf water volume and sapflux, we determined the turnover time of leaf water in the needles of Douglas-fir trees as approximately 11 h at midday. Such an extended turnover time suggests that transpiration may not have occurred at the commonly assumed isotopic steady state. We tested a non-steady state model for predicting delta 18O of leaf water. Our model calculations show that assuming isotopic steady state increased isoflux of transpiration. The impact of this increase on the modelled delta 18Ovp was clearly detectable, suggesting the importance of considering isotopic non-steady state of transpiration in studies of forest 18O water balance. PMID- 17086756 TI - Introduction of the carrot HSP17.7 into potato (Solanum tuberosum L.) enhances cellular membrane stability and tuberization in vitro. AB - We have examined the ability of a carrot (Daucus carota L.) heat shock protein gene encoding HSP17.7 (DcHSP17.7) to confer enhanced heat tolerance to potato (Solanum tuberosum L.), a cool-season crop. The DcHSP17.7 gene was fused to a 6XHistidine (His) tag to distinguish the engineered protein from endogenous potato proteins and was introduced into the potato cultivar 'Desiree' under the control of the cauliflower mosaic virus (CaMV) 35S promoter. Western analysis showed that engineered DcHSP17.7 was constitutively, but not abundantly, expressed in transgenic potato lines before heat stress. Leaves from multiple regenerated potato lines that contain the transgene exhibited significantly improved cellular membrane stability at high temperatures, compared with wild type and vector control plants. Transgenic potato lines also exhibited enhanced tuberization in vitro: under a condition of constant heat stress, at 29 degrees C, nodal sections of the transgenic lines produced larger and heavier microtubers at higher rates, compared to the wild type and vector controls. The dry weight and percentages of microtubers that were longer than 5 mm were up to three times higher in the transgenic lines. Our results suggest that constitutive expression of carrot HSP17.7 can enhance thermotolerance in transgenic potato plants. To our knowledge, this is the first study that shows that the thermotolerance of potato can be enhanced through gene transfer. PMID- 17086757 TI - Dynamics of water transport and storage in conifers studied with deuterium and heat tracing techniques. AB - The volume and complexity of their vascular systems make the dynamics of long distance water transport in large trees difficult to study. We used heat and deuterated water (D2)) as tracers to characterize whole-tree water transport and storage properties in individual trees belonging to the coniferous species Pseudotsuga menziesii (Mirb.) Franco and Tsuga heterophylla (Raf.) Sarg. The trees used in this study spanned a broad range of height (13.5-58 m) and diameter (0.14-1.43 m). Sap flow was monitored continuously with heat dissipation probes near the base of the trunk prior to, during and following injection of D2O. The transit time for D2O transport from the base of the trunk to the upper crown and the tracer residence time were determined by measuring hydrogen isotope ratios in water extracted from leaves sampled at regular intervals. Transit times for arrival of D2O in the upper crown ranged from 2.5 to 21 d and residence times ranged from 36 to 79 d. Estimates of maximum sap velocity derived from tracer transit times and path length ranged from 2.4 to 5.4 m d(-1). Tracer residence time and half-life increased as tree diameter increased, independent of species. Species-independent scaling of tracer velocity with sapwood-specific conductivity was also observed. When data from this study were combined with similar data from an earlier study of four tropical angiosperm trees, species-independent scaling of tracer velocity and residence time with sapwood hydraulic capacitance was observed. Sapwood capacitance is an intrinsic tissue-level property that appears to govern whole-tree water transport in a similar manner among both tracheid- and vessel-bearing species. PMID- 17086758 TI - Identification of QTL controlling root growth response to phosphate starvation in Arabidopsis thaliana. AB - One of the responses of plants to low sources of external phosphorus (P) is to modify root architecture. In Arabidopsis thaliana plantlets grown on low P, the primary root length (PRL) is reduced whereas lateral root growth is promoted. By using the Bay-0 x Shahdara recombinant inbred line (RIL) population, we have mapped three quantitative trait loci (QTL) involved in the root growth response to low P. The Shahdara alleles at these three QTL promote the response of the primary root to low P (i.e., root length reduction). One of these QTL, LPR1, located in a 2.8 Mb region at the top of chromosome 1, explains 52% of the variance of the PRL. We also detected a single QTL associated with primary root cell elongation in response to low P which colocalizes with LPR1. LPR1 does not seem to be involved in other typical P-starvation responses such as growth and density of root hairs, excretion of acid phosphatases, anthocyanin accumulation or the transcriptional induction of the P transporter Phtl;4. LPR1 might highlight new aspects of root growth that are revealed specifically under low P conditions. PMID- 17086759 TI - Growth at elevated CO2 concentrations leads to modified profiles of secondary metabolites in tobacco cv. SamsunNN and to increased resistance against infection with potato virus Y. AB - The effect of elevated CO2 concentrations on the levels of secondary metabolites was investigated in tobacco plants grown under two nitrogen supply (5 and 8 mM NH4NO3) and CO2 conditions (350 and 1000 p.p.m.) each. High CO2 resulted in a dramatic increase of phenylpropanoids in the leaves, including the major carbon rich compound chlorogenic acid (CGA) and the coumarins scopolin and scopoletin at both nitrogen fertilizations. This was accompanied by increased PAL activity in leaves and roots, which was even higher at the lower nitrogen supply. Hardly any change was observed for the structural phenolic polymer lignin and the sesquiterpenoid capsidiol. In contrast, elevated CO2 led to clearly decreased levels of the main nitrogen-rich constituent nicotine at the lower N-supply (5 mM NH4NO3) but not when plants were grown at the higher N-supply (8 mM NH4NO3). Inoculation experiments with potato virus Y (PVY) were used to evaluate possible ecological consequences of elevated CO2. The titre of viral coat-protein was markedly reduced in leaves under these conditions at both nitrogen levels. Since PR-gene expression and free salicylic acid (SA) levels remained unchanged at elevated CO2, we suggest that the accumulation of phenylpropanoids, for example, the major compound CGA and the coumarins scopolin and scopoletin may result in an earlier confinement of the virus at high CO2. Based on our results two final conclusions emerge. First, elevated CO2 leads to a shift in secondary metabolite composition that is dependent on the availability of nitrogen. Second, changes in the pool of secondary metabolites have important consequences for plant-pathogen interactions as shown for PVY as a test organism. PMID- 17086760 TI - Hydraulic redistribution in a Douglas-fir forest: lessons from system manipulations. AB - Hydraulic redistribution (HR) occurs in many ecosystems; however, key questions remain about its consequences at the ecosystem level. The objectives of the present study were to quantify seasonal variation in HR and its driving force, and to manipulate the soil-root system to elucidate physiological components controlling HR and utilization of redistributed water. In the upper soil layer of a young Douglas-fir forest, HR was negligible in early summer, but increased to 0.17 mm day(-1) (20-60 cm layer) by late August when soil water potential was approximately -1 MPa. When maximum HR rates were observed, redistributed water replenished approximately 40% of the water depleted from the upper soil on a daily basis. Manipulations to the soil or to the soil/plant water potential driving force altered the rate of observed HR indicating that the rate of HR is controlled by a complex interplay between competing soil and plant water potential gradients and pathway resistances. Separating roots from the transpiring tree resulted in increased HR, and sap flow measurements on connected and disconnected roots showed reversal of water flow, a prerequisite for HR. Irrigating a small plot with deuterated water demonstrated that redistributed water was taken up by small understorey plants as far as 5 m from the watering source, and potentially further, but the utilization pattern was patchy. HR in the upper soil layers near the watering plot was twice that of the control HR. This increase in HR also increased the amount of water utilized by plants from the upper soil. These results indicate that the seasonal timing and magnitude of HR was strongly governed by the development of water potential differences within the soil, and the competing demand for water by the above ground portion of the tree. PMID- 17086761 TI - Heart period variability in trauma patients may predict mortality and allow remote triage. AB - INTRODUCTION: The high frequency to low frequency ratio (HF/LF) derived from analysis of heart period variability is elevated and associated with mortality in severely injured patients monitored in a hospital. The purpose of this study was to test the utility of heart period variability measurements as indicators of injury severity in patients prior to definitive medical intervention. We tested the hypothesis that survival is associated with low relative HF/LF, and death is associated with high relative HF/LF. METHODS: We performed retrospective analyses of 84 pre-hospital trauma patient records (n=42 non-survivors; n=42 survivors) collected during helicopter transport to a Level 1 urban trauma center. R-waves from 2-min segments of ECG waveforms were converted to the frequency domain with a Fourier transform. Spectral power was separated into low (LF; 0.04-0.15 Hz) and high (HF; 0.15-0.4 Hz) frequency bands for analysis and derivation of frequency ratios. RESULTS: Absolute HF, LF, and HF/LF were not distinguishable statistically between groups (p > or = 0.26), but HF/LF was higher (p = 0.04) for non-survivors (140 +/- 26) than survivors (74 +/- 19). After normalization to account for large intersubject variability, HFnu (43 +/- 3 vs. 28 +/- 2) and HF/LFnu (248 +/- 50 vs. 73 +/- 19) were higher (both p < 0.001), and LFnu (42 +/- 4 vs. 64 +/- 3) was lower (p = 0.0001) for non-survivors [19 h (median) before death] compared with survivors. CONCLUSIONS: Our results show that heart period variability analyses separate patients who die from patients who survive traumatic injury. We propose that such analyses could be employed for remote triage of injured patients in austere environments. PMID- 17086762 TI - Melanoma growth and tumorigenicity in models of microgravity. AB - INTRODUCTION: Spaceflight involves numerous biological stressors that could affect long-term cancer incidence and tumor behavior. Ground-based models of microgravity can be used to investigate in vitro and in vivo tumor growth as a preparation for later work in space. The incidence of tumor growth and carcinogenesis in microgravity is as yet unknown. Hence, we investigated the effects of modeled microgravity on tumor growth and tumorigenicity using ground based in vitro and in vivo models. METHODS: Murine B16-F10 melanoma cells were cultured in a tissue culture flask (FL) and in a rotating-wall vessel bioreactor (BIO) designed by NASA to simulate some aspects of microgravity. We then measured cell growth, melanin production, and apoptosis. After 48 h of cultures in FL and BIO, cells were inoculated subcutaneously in C57BL/6 mice, syngeneic hosts for B16-F10 tumor cells. Tumor sizes were then measured every other day. RESULTS: BIO cultures had 50% decreases in growth when compared with FL cultures while demonstrating an inversely proportional increase in doubling time. Melanin production (a marker of differentiation) increased at 24 and 48 h in BIO. Flow cytometry analysis demonstrated that there was an increase in the percentage of apoptotic cells in the BIO when compared with that in the FL. When BIO-cultured melanoma cells were inoculated subcutaneously in mice, there was a significant increase in tumorigenicity as compared with FL-cultured cells. CONCLUSION: Our results indicate that simulated microgravity may have altered the tumor cell characteristics and enhanced the invasive property. It is possible that the microgravity analogue culture environment may have selected highly tumorigenic cells for survival despite the decreased overall growth in the microgravity analogue. PMID- 17086763 TI - Gravity replacement during running in simulated microgravity. AB - INTRODUCTION: During treadmill exercise on the International Space Station (ISS), a restoring load from a subject load device (SLD) is applied through a shoulder and-waist harness to pull the exercising crewmember toward the treadmill surface. The capacity of this arrangement to provide 1-g-like reaction forces may be critical for effective use of the treadmill as a countermeasure to musculoskeletal changes during prolonged spaceflight. This study in simulated microgravity evaluated the comfort and function during running of the ISS harness used with a new SLD in a system that allows more even distribution of the load between the waist and shoulders. METHODS: Using a zero-gravity locomotion simulator, 12 subjects completed three 5-min running trials at a constant speed (3.35 m x s(-1)) using three SLD loads [50%, 75%, and 100% of bodyweight (BW)] presented at random and a shoulder-to-waist loading ratio of 50:50. Subjective ratings of discomfort, ground reaction forces (GRFs), and SLD loads were collected. RESULTS: A load of 100% BW resulted in similar GRF profiles (peak and rate of change of force) to those reported for 1-g running over ground and were also comfortably tolerated (mean Borg scale rating 3.9/10). DISCUSSION: With an appropriate harness and SLD system, 1-g-like GRF profiles can be generated at the feet during simulated microgravity running. Such forces can be achieved with a level of discomfort rated better than "somewhat uncomfortable," suggesting that running with 1-g loads could be an effective component of musculoskeletal countermeasures during long-duration spaceflight. PMID- 17086764 TI - Squat-stand test response following 10 consecutive episodes of head-up tilt. AB - INTRODUCTION: The cardiovascular system is adaptable to multiple exposures to gravity over several days, and to repeated exposures in a single day. This study aimed to investigate if the cardiovascular adaptation observed following 10 +75 degrees head-up tilts (HUT) would improve the responses to the squat-stand test (SST). METHODS: There were 16 subjects who were randomly allocated into either a tilting group that underwent 10 +75 degrees HUTs in 70 min (Tilting) or a control group that underwent 70 min of rest (Control). Before and after HUT or rest, subjects performed a SST (SST1 and SST2, respectively). Heart rate (HR), BP [systolic (SBP), diastolic (DBP), mean (MAP)], total peripheral resistance (TPR), stroke volume (SV), and cardiac output (CO) were determined during both SSTs. The final 30 s of squat and the first 30 s of stand (divided into three 10-s blocks termed Stand10, Stand20, and Stand30) were compared between SST1 and SST2. RESULTS: In the Tilting condition, during the squat phase of SST2, the following were significantly elevated: SBP (131 +/- 9 to 140 +/- 7.2 mmHg) and MAP (94 +/- 8.7 to 105 +/- 10.2 mmHg); DBP (76 +/- 9.4 to 87 +/- 11.9 mmHg); TPR (1197 +/- 524.6 to 229 +/- 610.5 dyn x s(-1) x cm(-5)) and HR were significantly decreased (78 +/- 6.9 to 73 +/- 7.5 bpm) compared with SST1. At Stand10, DBP and MAP were significantly increased (59 +/- 9.5 to 69 +/- 15.7 mmHg and 74 +/- 8.9 to 84 +/- 14.7 mmHg, respectively); at Stand20, SBP was increased (121 +/- 17 to 128 +/- 22 mmHg); and at Stand30 the following were increased: DBP (64 +/- 8.8 to 75 +/- 16.1 mmHg); SBP (127 +/- 9.2 to 139 +/- 15.8 mmHg); and MAP (79 +/- 8 to 90 +/- 14.9 mmHg). There were no differences observed between SST1 and SST2 in the Control group. DISCUSSION: Cardiovascular responses to the SST can be improved with 10 consecutive +75 degrees HUTs. This is predominantly due to an increase in DBP, indicative of a change in vascular resistance. PMID- 17086765 TI - Cervical resistance training: effects on isometric and dynamic strength. AB - INTRODUCTION: Neck injuries signify a physical fitness and human system problem with high operational significance. The prevalence of injuries in tactical aviators has been reported to be as high as 84%, although few report engaging in neck-specific strengthening exercises. It is generally believed that neck strengthening may result in fewer neck injuries. This study was designed to investigate the effects of 12 wk of cervical strength training (3 d x wk(-1)) on isometric strength, dynamic strength, and hypertrophy in a sample of military men. METHODS: Participants were tested for each of the above-mentioned variables before and after the training program as well as at 4-wk intervals, and results were compared with a control group that performed no cervical resistance training. RESULTS: Results indicated significant improvements in isometric strength and dynamic strength, typically occurring as early as 4 wk and improving throughout the 12-wk period. Modest increases in neck circumference were also noted. DISCUSSION: These findings have implications for military personnel at risk of neck injury in their occupational activities. PMID- 17086766 TI - Breath-hold performance during cold water immersion: effects of psychological skills training. AB - INTRODUCTION: Accidental cold water immersion (CWI) is a significant cause of death, particularly in those who are immersed in rough water or forcibly submerged such as in a ditched and inverted helicopter. The marked reduction in maximal breath-hold time associated with CWI, part of the 'cold shock' response, significantly increases the risk of drowning. However, the response is highly variable between subjects. This experiment tested the hypothesis that part of this variability is due to psychological factors. METHODS: There were 32 subjects who completed 2 2.5-min, head-out immersions in 11 degrees C water, separated by 7 d. Between immersions, subjects were matched on initial maximum breath-hold time on immersion (BHwater) and allocated to either a psychological intervention group (PIG) or control group (CG). PIG (n=16) subjects each undertook a psychological skills intervention comprising 4 interlinked training sessions covering goal-setting, arousal regulation, mental imagery, and positive self talk; CG (n=16) continued normal daily activity. RESULTS: Psychological intervention significantly increased BHwater on immersion in the PIG vs. the CG [mean (SD); CG BHwater immersion 1:24.01 (6.72) s; immersion 2: 21.34 (16.31) s; PIG: BHwater immersion 1: 24.66 (14.60) s; immersion 2: 44.25 (31.63) s]. The difference in maximum voluntary BHwater between immersion 1 and 2 in the PIG averaged 19.59 s, equating to an 80% increase following psychological intervention. CONCLUSION: Psychological influences may account for a significant amount of the variability in the respiratory responses during CWI, and may be a key factor in determining the chances of survival following accidental immersion. PMID- 17086767 TI - Normobaric hypoxia inhalation test vs. response to airline flight in healthy passengers. AB - INTRODUCTION: There is little data available to determine the normal response to normobaric hypoxia inhalation testing (NHIT) and air travel. Quantifying a healthy response may assist in the evaluation of passengers considered at risk for air travel. The aims of this study were: (1) to quantify the degree of desaturation in healthy subjects during a NHIT and air travel; and (2) assess the validity of the NHIT when compared with actual in-flight responses. METHODS: There were 15 healthy adults (age 23-57; 10 women) who volunteered for this study. Preflight tests included lung function, arterial blood gas, pulse oximetry (SpO2), and NHIT (inspired oxygen 15%). SpO2 and cabin pressure were measured continuously on each subject during a commercial air flight (mean cabin altitude 2178 m; range 1719-2426 m). In-flight oxygenation was compared with the preflight NHIT. RESULTS: Lung function testing results were normal. There was significant desaturation (SpO2) during the NHIT (pre: 98 +/- 2%; post: 92 +/- 2%) and at cruising altitude (pre: 97 +/- 1%; cruise: 92 +/- 2%). There was no difference between the final NHIT SpO2 and the mean in-flight SpO2. There was a significant difference between the lowest in-flight SpO2 (88 +/- 2%) vs. the lowest NHIT SpO2, (90 +/- 2%). DISCUSSION: Oxygen saturation decreases significantly during air travel in normal individuals. In this group of healthy passengers the NHIT approximates some, but not all, aspects of in-flight oxygenation. These results can be used to describe a normal response to the NHIT and air-travel. PMID- 17086768 TI - Genetic influences on motion sickness susceptibility in adult women: a classical twin study. AB - BACKGROUND: Motion sickness is a common and potentially debilitating condition that characteristically occurs in situations of conflicting sensory input. While the precise stimuli that give rise to this trait are increasingly well characterized, the underlying determinants of individual susceptibility to motion sickness remain unclear. This study uses a classical twin design to assess the influence of genetic and environmental factors. METHODS: A postal survey was conducted in an age-matched sample of 3652 monozygotic (MZ) and dizygotic (DZ) adult female twins selected from the TwinsUK Registry. Study participants were asked to complete items from a validated questionnaire relating to their lifetime susceptibility to motion sickness. The relative contribution of genetic and environmental factors to motion sickness susceptibility was assessed using variance components analysis. RESULTS: The response rate to the questionnaire was 78%. Approximately 40% of respondents reported at least moderate susceptibility to motion sickness. The pattern of responses among twins indicated a significant genetic contribution with heritability for a motion sickness factor score estimated as 57% (95% CI: 51%, 63%). The heritability of recalled motion sickness was at its highest in childhood (70% [59%, 80%]) and declined through puberty and the early adult years. DISCUSSION AND CONCLUSIONS: The findings highlight the importance of genetic factors in determining an individual's underlying propensity to motion sickness and should stimulate the search for specific susceptibility genes. PMID- 17086769 TI - CNS oxygen toxicity in closed-circuit diving: signs and symptoms before loss of consciousness. AB - INTRODUCTION: There is a dearth of information regarding CNS oxygen toxicity accidents in closed-circuit oxygen diving. The aims of the present study were to report the sensations and symptoms that accompany CNS oxygen toxicity accidents, and to evaluate whether loss of consciousness can occur without any warning signs. METHODS: We documented 36 CNS oxygen toxicity accidents in closed-circuit oxygen diving. The full accident inquiry included the first report from the diving unit, an interview of the victim and his buddy by the researchers, and an examination of the diving equipment. RESULTS: The symptoms that appeared before termination of a dive, as reported by the victim or his buddy, were as follows (in descending order of frequency): limb convulsions; hyperventilation; difficulty maintaining a steady depth; headache; and visual disturbances. The symptoms that appeared after detachment from the mouthpiece were, in descending order of frequency: headache; loss of consciousness; confusion; weakness; dizziness; and facial muscle twitching and limb convulsions. A high inspired CO2 [mean 4.2 kPa (29.9 mmHg)] was connected with loss of consciousness. No dive was terminated before at least two symptoms (mean 3.4) had been noted a minimum of 5 min before termination. DISCUSSION: Symptoms that are accepted as being related to CNS oxygen toxicity, as well as others such as headache, difficulty maintaining a steady depth, hyperventilation, weakness, and a choking sensation, were more frequent among the O2 accident victims compared with divers who did not interrupt their dives. CONCLUSION: Awareness of any unusual sensation can prevent a potentially dangerous situation from arising. PMID- 17086770 TI - Cervical and lumbar MRI findings in aviators as a function of aircraft type. AB - BACKGROUND & AIMS: Neck pain and lower back pain (LBP) are frequently reported by military helicopter pilots (HP) and fighter pilots. A small number of studies have used imaging methods to evaluate spinal cervical degenerative findings in pilots exposed to high +Gz, with results indicating an increase in cervical disk protrusions in this population. We evaluated the cervical and lumbar spine with magnetic resonance imaging (MRI) to assess the prevalence of degenerative changes in three subpopulations of pilots. METHODS: Fighter pilots (FP), transport pilots (TP), and HP (10 pilots in each group) underwent cervical and lumbar MRI. Degenerative pathologic changes (disk herniation, cord compression, foraminal stenosis, and the presence of osteophytes) were evaluated in each group by two independent experienced radiologists. RESULTS: Cervical spine degenerative changes seemed to be associated with older age rather then aircraft type, affecting the older group of TP (8/10 pilots) more than the younger FP group who were exposed to high +Gz (3/10 pilots). In contrast, for lumbar spine degenerative changes, we found an uncommon pattern of lumbar spine degeneration in HP, affecting the upper part of the lumbar spine (10/13 disks found at L1-L4). CONCLUSIONS: The results of this study suggest that HP may have detectable degenerative lumbar findings. More research is needed to validate these findings as well as to explore the possible pathophysiological link between occupational exposures and the specific involvement of the upper lumbar spine. PMID- 17086771 TI - Pre-hypertension as a predictor of hypertension in military aviators: a longitudinal study of 367 men. AB - BACKGROUND: Recently, the seventh report of the Joint National Committee on prevention, detection, evaluation, and treatment of high blood pressure introduced the term "pre-hypertension" for systolic blood pressure (BP) levels of 120-139 and diastolic BP levels of 80-89 mmHg. Our aim was to establish the prevalence of this novel entity among young subjects, and to assess the rate of progression to hypertension. METHODS: We retrospectively analyzed the data of a group of 367 male aviators who were examined at the age of 18 and who were followed for an average of 18 +/- 7 yr (range 3-37 yr). Systolic and diastolic BP, heart rate, and body mass index (BMI) were measured at the age of 18 and at the end of the follow-up. RESULTS: At the age of 18, 176 subjects (48%) had pre hypertension. Subjects with pre-hypertension had higher BP levels (132 +/- 6/76 +/- 8 vs. 115 +/- 6/72 +/- 7 mmHg; p < 0.001), heart rate (71 +/- 12 vs. 65 +/- 12 bpm; p < 0.001) and BMI (22.4 +/- 2.9 vs. 21.7 +/- 2.5 kg x m(-2); p < 0.05) than the subjects with normal BP. During the follow-up period, 110 subjects (30%), 77 with pre-hypertension and 33 with normal BP, developed hypertension. Pre-hypertensive subjects had an odds ratio of 3.7 (95% CI; 2.3-6.2) to develop hypertension as compared with subjects with normal BP. CONCLUSIONS: Pre hypertension is common and it predicts the development of frank hypertension. Thus, subjects with pre-hypertension should be closely followed and encouraged to begin lifestyle modification in order to prevent future complications. PMID- 17086772 TI - Fatigue in military aviation shift workers: survey results for selected occupational groups. AB - BACKGROUND: Shift workers are particularly vulnerable to increased sleepiness, chronic fatigue, and decreased performance, which can adversely impact productivity and safety in military flight operations. This study examined the association of specific risk factors including work context and shift system details (squadron: remotely piloted aircraft [RPAI vs. manned aircraft [MA]), work/rest guidelines (career field: crewmember vs. maintainer), and participation in deployed operations (environment: home base vs. deployed) on subjective fatigue using standardized and validated fatigue questionnaires. METHODS: A cross sectional survey of 172 U.S. Air Force (USAF) personnel was conducted from October 2004 to May 2005. The study sample was recruited from four different USAF occupational groups involved in some form of shift work to include irregular, rotational, or fixed shifts. RESULTS: Participants reported a mean (SD) of 6.6 (1.8) hours of sleep per day with no differences by squadron, career field, or environment. Mean daily sleep did not correlate with scores on the fatigue questionnaires. Mean scores on the fatigue questionnaires were associated with squadron (mean fatigue score: RPA > MA), but not with career field or environment. There were no significant interaction effects, nor were there significant effects based on the covariates age, gender, and rank. CONCLUSION: Work context, shift system details, or both appeared to best explain the observed differences in fatigue between USAF shift worker populations. Crewmember work/rest guidelines did not appear to be useful for mitigating fatigue associated with shift work. Shift work is intrinsically fatiguing, regardless of whether the shift worker is at home base or deployed. PMID- 17086773 TI - Pilot medical history and medications found in post mortem specimens from aviation accidents. AB - INTRODUCTION: Federal Aviation Administration (FAA) regulations require pilots to report all medications and medical conditions for review and consideration as to the overall suitability of the pilot for flight activities. METHODS: Specimens were collected by local pathologists from aviation accidents and sent to the Bioaeronautical Sciences Research Laboratory for analysis. The results of such tests were entered into the Forensic Case Management System. This database was searched to identify all pilots found positive for medications used to treat cardiovascular, psychological, or neurological conditions over the period January 1, 1993, through December 31, 2003. RESULTS: Toxicological evaluations were performed on 4143 pilots. Psychotropic drugs were found in 223 pilots. Cardiovascular medications were found in 149 pilots. Neurological medications were found in 15 pilots. Pilots reported psychological conditions in 14 of the 223 pilots found positive for psychotropic drugs. Only 1 of the 14 pilots reporting a psychological condition to the FAA reported the psychotropic medication found after the accident. Cardiovascular disease was reported by 69 of the pilots found with cardiovascular drugs in their system. Cardiovascular medications found in the pilots were reported by 29 of the 69 pilots reporting a cardiovascular condition. Only 1 of the 15 pilots reported having a neurological condition to the FAA; none of the pilots found with neurological medications reported the medication. CONCLUSIONS: Toxicology successfully identified 93% of the medications reported by the pilots. Pilots involved in fatal accidents taking psychotropic or neurological medications rarely reported the medication or their underlying medical condition to the FAA. PMID- 17086774 TI - Thermoregulatory control following dynamic exercise. AB - Post-exercise thermoregulatory control in humans has received limited attention. In the post-exercise period skin blood flow and sweating return to pre-exercise levels despite a persistent elevation in esophageal temperature, suggesting an alteration in thermoregulatory control. The esophageal temperature response post exercise appears to be correlated to the marked cardiovascular changes that occur after dynamic exercise. Recent work has shown that non-thermoregulatory factors associated with hemodynamic changes and hydration status post-exercise may influence the regulation of core temperature during exercise recovery. This review will characterize the thermal response and describe our current understanding of the physiological influences on thermoregulatory control during recovery from dynamic exercise. PMID- 17086775 TI - Immersion in thermoneutral water: effects on arterial compliance. AB - BACKGROUND: Since thermoneutral head-out water immersion induces an increase in central blood volume and cardiac output along with a decrease in peripheral vascular resistance, we examined here whether thermoneutral immersion leads rapidly to an increase in arterial compliance. METHODS: Changes in hemodynamic status and arterial compliance induced by thermoneutral head-out water immersion to the midchest were studied in 26 healthy volunteers (15 women, 11 men) using a non-invasive pulse wave measurement system. Several hemodynamic parameters including stroke volume (SV) and large and small artery compliance were studied in a quiet room with a stable environmental temperature (25 degrees C) and during thermoneutral (34.5 degrees C) head-out water immersion. RESULTS: In a preliminary study, an agreement between SV estimated by radial pulse contour analysis and that obtained by Doppler echocardiography was observed in ambient air. An increase in SV led to an increase in cardiac output (12%) 10 min after the start of immersion. Heart rate and arterial pressure remained unchanged. Systemic vascular resistance was significantly decreased and total arterial compliance estimated by the SV to aortic pulse pressure ratio was significantly increased. Radial pulse contour analysis confirmed these results with an increase in both large and small artery compliance. CONCLUSION: In conclusion, radial pulse contour analysis provides an interesting method for assessing hemodynamic changes under environmental constraints. Our results were consistent with a prompt increase in arterial compliance during acute water immersion. PMID- 17086776 TI - The role of aeronautical adaptability in the disqualification of a military flyer. AB - INTRODUCTION: Possessing a stable personality plays a critical role in crew coordination, mission completion, and safety of flight in aviation. Presented is a naval flight officer whose personality traits were a threat to these key tenets and ultimately warranted removal from flight status. CASE REPORT: A naval flight officer was twice referred to Human Factors Boards (HFBs) after concerns were raised about her emotional stability in and out of the cockpit. Her history revealed multiple clinical presentations revealing personality characteristics that should have raised immediate concern about her aeronautical adaptability. Formal psychiatric evaluation after her second HFB confirmed the presence of personality traits incompatible with aviation duty, but not until she had served nearly 4 yrs. as an aviation officer. DISCUSSION: Aeronautical adaptability is a U.S. Naval term used to describe an individual's ability to work successfully in the aviation environment. Naval aviation personnel are considered not aeronautically adaptable/adapted if diagnosed with a "personality disorder or prominent personality traits that adversely affect flight safety, mission completion, or crew coordination". The Federal Aviation Administration, the U.S. Air Force, and the U.S. Army endorse similar concepts. Except for the U.S. Navy definition, aeronautical adaptability does not include formal DSM-IV-TR diagnoses, and, thus, flight surgeons and aviation medical examiners often make difficult aeromedical dispositions based on vague and subjective criteria. CONCLUSIONS: Determining aeronautical adaptability is not a simple, solitary process. It requires heightened suspicion, continual surveillance of suspect aviators, and collaboration from aviation medical professionals to ensure aviation safety. PMID- 17086777 TI - Smallpox--inappropriate vaccination site. AB - This case reviews the current literature concerning smallpox and the smallpox vaccination program. Site placement of the vaccine had the unexpected effect of grounding an airman for an extended period of time. PMID- 17086778 TI - You're the flight surgeon. Lumbar radiculopathy. PMID- 17086779 TI - Parachute opening shock emulator with pitch and yaw control. PMID- 17086780 TI - The Civil Aviation Medical Association's history. PMID- 17086781 TI - Factor analysis of the Revised Piper Fatigue Scale in a caregiver sample. AB - The Revised Piper Fatigue Scale (R-PFS) is an instrument designed to measure subjective fatigue that was developed in samples with physical illness. Its psychometric properties in nonclinical samples are unknown. The purpose of this study was to examine the psychometric properties of the R-PFS in a sample of caregivers of stroke survivors. The convenience sample of 132 caregivers was primarily women (74%), White (71%), college-educated (73%), and employed (52%), with a mean age of 56.7 years (SD = 13.71). Internal consistency reliabilities for the four R-PFS subscales and the total scale were excellent, ranging from .90 to .97. Principal axis factor analysis with oblique rotation was conducted to examine construct validity of the R-PFS. A three-factor solution explained 75.9% of the common variance. Two factors totally replicated the behavioral/severity and affective meaning subscales of the R-PFS. The third factor incorporated a combination of Piper's sensory and cognitive/mood subscales and appeared to summarize how fatigue makes the caregiver feel. The R-PFS demonstrated strong internal consistency reliability and construct validity in this sample. However, data suggest that caregivers may perceive certain feelings associated with fatigue as conceptually similar when these feelings are conceptually distinct in Piper's breast cancer sample. The study supports the need for psychometric evaluation of instruments developed in clinical populations prior to their use in nonclinical populations. PMID- 17086782 TI - Comparison of three modes of measuring stress, coping, and humor in school-age children. AB - Computer-assisted administration of surveys is gaining popularity among many researchers, but the equivalence of this method to more traditional approaches such as using paper and pencil has not been determined for many commonly used questionnaires, particularly among school-age children. This study examined systematic differences in the responses of 4th, 5th, and 6th graders to measures of stress, coping, and humor among three modes of assessment: paper-and-pencil questionnaires, computer-assisted self-interviewing (CASI), or a combination of paper-and-pencil and CASI. Participants were 1,245 ethnically diverse children enrolled in public schools in the central region of the United States. Psychometric and score distribution characteristics were examined using item analyses and analyses of mean and covariance structure as a function of mode of assessment. Differences in response patterns, primarily at the scale score level, were documented on some of the key measures. In general, CASI medians and means were higher and correlations among CASI measures tended to be lower than those obtained with paper-and-pencil and mixed mode assessment, and CASI variances were lower. This study suggests the importance of the continued examination of the impact of mode of questionnaire administration when assessing these and other domains of well-being in school-age children. PMID- 17086783 TI - Development of a culturally specific instrument for mammography screening: an example with American Indian women in Vermont. AB - This article presents the triangulation process for translating qualitative data about mammography screening from a grounded theory study with American Indian women in Vermont, into questionnaire items based on an existing model of behavior change, the Transtheoretical Model (TTM) Stages-of-Change. Qualitative data were used to derive a theory, Moving in Between Mammography, which suggested that traditionality influenced American Indian women's screening decisions. To examine the relationship between mammography and traditionality, new items were developed for each of three key TTM constructs: Pros, Cons, and Processes-of-Change. The process for developing the new TTM-based items, as well as traditionality items specific for American Indian women living off-reservation, are presented. This article provides one example of how an instrument can be developed within a culturally competent nursing framework. PMID- 17086784 TI - Daily events and mood state among individuals living with HIV: examination of the within-persons approach to data collection using daily diary methodology. AB - Studies examining the link between stressful events and coping with HIV have relied on a between-persons approach focusing on how individuals differ on some characteristics. Although the between-subjects approach has yielded important information, our goal was to use a within-persons approach, making repeated measurements of the same persons over many days, to examine the impact of changing circumstances on the mood states of those with HIV. A second goal was to determine if asking participants to report their daily experiences via a computerized interactive voice system is a viable way to collect such information. This study collected a variety of trait measures for seven HIV patients and subsequently used a computerized telephone system to collect information regarding daily events and mood states over 21 consecutive days. Several daily measures, including self-esteem, optimism, and positive social interactions were significantly related to daily mood states. Trait measures, with the exception of symptom distress, were ineffective in predicting variations in daily mood states. It was concluded that a computerized telephone system is a viable means of collecting information from HIV patients on a daily basis and within-persons methodology may provide useful information about daily events affecting mood states beyond that generated by a between-persons approach. PMID- 17086785 TI - Field testing, refinement, and psychometric evaluation of a new measure of nursing home care quality. AB - The primary aim of this NINR-NIH-funded field test in 407 nursing homes in 3 states was to complete the development of and conduct psychometric testing for the Observable Indicators of Nursing Home Care Quality Instrument (Observable Indicators, OIQ). The development of the OIQ was based on extensive qualitative and iterative quantitative work that described nursing home care quality and did initial validity and reliability field testing of the instrument in 123 nursing homes in 1 state. The scale is meant for researchers, consumers, and regulators interested in directly observing and quickly evaluating (within 30 minutes of observation) the multiple dimensions of care quality in nursing homes. After extensive testing in this study, the Observable Indicators instrument has been reduced to 30 reliable and discriminating items that have a conceptually coherent hierarchical factor structure that describes nursing home care quality. Seven first-order factors group together into two second-order factors of Structure (includes Environment: Basics and Odors) and Process (includes Care Delivery, Grooming, Interpersonal Communication, Environment: Access, and Environment: Homelike) that are classic constructs of Quality, which was the third-order factor. Internal consistency reliability for the 7 first-order factors ranged from .77 to .93. Construct validity analyses revealed an association between survey citations and every subscale as well as the total score of the OIQ instrument. Known groups analysis revealed expected trends in the OIQ scores. The Observable Indicators instrument as a whole shows acceptable interrater and test retest reliabilities, and strong internal consistency. Scale subscales show acceptable reliability as well. Generalizability Theory analyses revealed that dependability of scores can be improved by including a second site observer, or by revisiting a site. There is a small additional benefit from increasing observers or visits beyond two. PMID- 17086786 TI - Organizational ethics: why bother? PMID- 17086787 TI - New resource aids discernment of the critical relationships in sponsorship. PMID- 17086788 TI - Congressional conversations: horse slaughtering or poverty and health care? PMID- 17086789 TI - A wondrous history of community benefit. PMID- 17086790 TI - P3 in action. PMID- 17086791 TI - "Like shining from shook foil". A "virtuous organization" is prepared to treat both the body and the soul. AB - The Catholic health care ministry is about mission, and the role of organizational ethical reflection is to encourage people in the ministry to think about the institutional performance and practice of medicine within a ministry of the Catholic Church. By engaging a creative process that identifies the needs of people served by Catholic health care, institutions are able to mediate the healing and redeeming power of Jesus, thereby creating virtuous organizations. To depict the mission of Catholic health care as an extension of the healing ministry of Jesus is to evoke explicitly Catholic theological language, and such language is appropriate because Catholic health care is a ministry of the Catholic Church. The church itself is the embodiment of the healing and redeeming ministry of Jesus, and the institutional ministries it has created over time need to bear witness to this fundamental reason for their existence. PMID- 17086792 TI - Finding a voice. Like individuals, organizations are moral speakers and actors. AB - Though "good people' are important for the life of any organization, it is a myth to think that enough good people will make for a good organization. To break free of this myth, a health care organization, which is made up of numerous persons and groups, ought to be regarded as a single, unitary actor in society. When seen as a single actor, the organization's systems for carrying out its mission can be better assessed and improved if necessary. If the organization's systems are not functioning as they should, then even good people will be hindered in their efforts. It can be said, therefore, that organizational ethics takes seriously the idea that every Catholic health care organization is a moral actor needing to reflect carefully on what it does in relation to its employees, leaders, and the outside community. In an environment where the organization's actions are reflected upon, and its character is carefully and continually shaped according to its mission, individual persons in that organization will be better equipped for making and carrying out good decisions that are aligned with that same regard for the mission. PMID- 17086793 TI - Shedding light on organizational ethics. Five ethicists help define and contextualize an elusive topic. PMID- 17086794 TI - Starting an organizational ethics committee. An ethicist suggests some practical and concrete steps. AB - Establishing an organizational ethics committee (OEC) involves careful reflection on the needs of the organization and on the people who will serve on the committee. With concern for the "community of care" (the women and men who carry out the organization's mission), a comprehensive needs assessment will reveal areas of the organization where more education and policy analyses are needed. Volunteer members of the OEC are typically chosen according to a set of characteristics that include their knowledge and experience, ability to take on this added responsibility, familiarity with the Ethical and Religious Directives for Catholic Health Care Services and Catholic social justice teaching, and their honesty and integrity. Part of the success of an OEC can be attributed to how well it is supported by the organization. This includes administrative and financial support, use of public relations and educational services, and cooperation and active involvement of key managers in the organization. Once formed, the next important step is educating OEC members. PMID- 17086795 TI - Hiring for "organizational fit". A St. Louis system has developed tools for assessing candidates' compatibility with the organization. AB - To a great extent, the continued success of Catholic health care organizations is dependent on the selection of co-workers and leaders who are committed to carrying on the organization's mission. The Sisters of Mercy Health System, St. Louis, uses three tools to help leaders be more consistent and objective in assessing employment candidates for organizational fit. The first tool involves behavioral-based interviewing, which looks at a candidate's potential for future behaviors based on his or her past behaviors. The second tool assesses a candidate's values in such a way that accounts for and reduces the interviewer's own subjectivity. And the third tool-values-based interviewing-helps reveal a candidate's natural aptitude for, interest in, and personal agreement with the organization's charism, religious heritage, and values. PMID- 17086796 TI - Bridging the gap. Catholic health care organizations need concrete ways to connect social principles to practice. AB - Establishing and maintaining institutional identity is a challenge for leaders in Catholic health care. A process known as "progressive articulation" can be used to help leaders assess how well their organizations reflect Catholic social tradition and help them apply this tradition toward specific organizational practices. The particular approach described here is called the "Identity Inquiry and Improvement Process" (31P), and it takes Catholic social principles and translates them into criteria and benchmarks for assessing an organization's interactions with internal and external stakeholders. In other words, 31P seeks to make mission measurable and concrete. PMID- 17086797 TI - Managing change. Wheaton Franciscan Healthcare has developed an interdisciplinary approach. PMID- 17086798 TI - Looking back on Catholic Healthcare West. A sponsor's vision for Catholic health care. PMID- 17086799 TI - Nursing and the common good. A clearer definition of the concept could be helpful to all the healing professions. PMID- 17086800 TI - Saving lives at Centura. A Colorado System has developed a rapid-response team for critically ill patients. PMID- 17086801 TI - "Formation" in the Code of Canon Law. PMID- 17086802 TI - Gestational trophoblastic disease in ectopic pregnancy: A case series. AB - OBJECTIVE: To describe 6 cases of gestational trophoblastic disease (GTD) in ectopic pregnancy admitted to Hospital Universitario de Caracas (HUC). STUDY DESIGN: Medical records of 6 patients admitted to the Obstetrics and Gynecology Department, HUC, from 1996 to 2004 were reviewed. They underwent surgery with a diagnosis of ectopic pregnancy, and histologic analysis revealed GTD. Clinical trends were analyzed. RESULTS: The prevalence of GTD in ectopic pregnancy was 0.16:1,000 deliveries. The mean patient age was 29 years. The preceding gestation was a term delivery in 4 and abortion in 2. The mean gestational age at admission was 8 weeks. All patients complained of abdominal pain, and 3 of them also had vaginal bleeding. Ultrasound revealed an adnexal tumor in 5 cases; this tumor and hemoperitoneum (6 cases) were the most frequent surgical findings. Histopathologic diagnosis was partial mole in 5 and choriocarcinoma in 1. Four patients were lost to follow-up. CONCLUSION: In this series the prevalence of ectopic GTD was high. The condition can mimic the usual symptoms of ectopic pregnancy, especially when a hemoperitoneum is present. It is important to apply strict histologic criteria for GTD when a sample of ectopic pregnancy is analyzed and to monitor those patients with careful human chorionic gonadotropin follow up. PMID- 17086803 TI - Persistent gestational trophoblastic neoplasia after partial hydatidiform mole incidence and outcome. AB - OBJECTIVE: To report the Sheffield experience with persistent gestational trophoblastic neoplasia (GTN) after partial hydatidiform mole (PHM) and to review worldwide experience. METHODS: All PHMs registered at the Sheffield Trophoblast Centre between 1991 and 2004 were included in this retrospective study. Any case of PHM leading to persistent GTN was reviewed centrally by an expert gynecologic pathologist. Clinical features, treatment and outcome were recorded. RESULTS: During the 14-year study period 3189 PHMs were registered. Forty-one developed persistent GTN. Central histopathology review confirmed PHM in only 14 cases (0.91% of all those registered). Twelve scored low and 2 high risk according to International Federation of Gynecology and Obstetrics 2000 criteria. During the same period, 271 cases of persistent GTN originally registered as complete hydatidiform mole were reviewed; 3 were found to be PHMs (2 low, 1 high risk). In all, 15 of 17 persistent PHMs required chemotherapy. CONCLUSION: Persistent GTN requiring chemotherapy can occasionally occur after PHM; surveillance of all cases continues to be recommended. PMID- 17086804 TI - Primary treatment of metastatic high-risk gestational trophoblastic neoplasia with EMA-CO chemotherapy. AB - OBJECTIVE: To evaluate the efficacy of etoposide, methotrexate, actinomycin D, cyclophosphamide and vincristine (EMA-CO) in the primary treatment of metastatic high-risk gestational trophoblastic neoplasia. STUDY DESIGN: Thirty women with metastatic high-risk gestational trophoblastic neoplasia were treated primarily with EMA-CO between 1986 and 2005. Patients who had incomplete responses or developed resistance to EMA-CO were treated with drug combinations employing etoposide and a platinum agent with or without bleomycin or ifosfamide. Adjuvant surgery and radiotherapy were used in selected patients. Survival, clinical response and factors affecting treatment success were analyzed retrospectively. RESULTS: The overall survival rate was 93.3% (28 of 30). Of the 30 patients treated with EMA-CO, 20 (66.7%) had a lasting clinical response, 8 (26.7%) developed resistance but were subsequently placed in remission with platinum based chemotherapy, and 2 (6.7%) died of widespread metastatic disease. Clinical complete response to EMA-CO was significantly influenced by human chorionic gonadotropin level (<100,000 mIU/ mL, 82%, vs. > 100,000 mIU/mL, 46%), metastatic site (lung and pelvis, 75%, vs. other, 33%) and International Federation of Gynecology and Obstetrics (FIGO) risk factor score (< 7, 92% vs. >7, 50%). Surgical procedures were performed on 12 patients, and 4 patients received brain irradiation. Eight (80%) of 10 patients who received secondary platinum-based chemotherapy or without surgery were cured. The 2 patients who died had stage IV disease (brain and/or liver metastases) with FIGO scores of 13 and 14. CONCLUSION: Over 93% of 30 patients with metastatic high-risk gestational trophoblastic neoplasia treated initially with the EMA-CO protocol, often in conjunction with brain irradiation, surgical resection of sites of persistent tumor and salvage platinum-based chemotherapy, were cured. PMID- 17086805 TI - Role of surgery in the management of high-risk gestational trophoblastic neoplasia. AB - OBJECTIVE: To evaluate the role of surgery in the management of high-risk gestational trophoblastic neoplasia. STUDY DESIGN: Twenty-four (48%) of 50 patients treated with etoposide, methotrexate, actinomycin D, cyclophosphamide and vincristine (EMA-CO) regimen as primary or secondary chemotherapy for high risk gestational trophoblastic neoplasia between 1986 and 2005 underwent 28 adjuvant surgical procedures. The procedures included hysterectomy (17), lung resection (5), salpingectomy (1), uterine wedge resection (1), small bowel resection (1), suturing of the liver or uterus for bleeding (2) and uterine artery embolization (1). RESULTS: Twenty-one (87.5%) of 24 patients who had surgical procedures as part of their treatment for high-risk disease survived. Fifteen (88%) of 17 patients undergoing hysterectomy were cured. Four (80%) of 5 patients who had resistant foci of choriocarcinoma in the lung were cured by pulmonary resection. The patients who had suturing of the uterus, uterine artery embolization, small bowel resection and salpingectomy for bleeding as well as the patient who had uterine wedge resection of resistant choriocarcinoma survived. CONCLUSION: Adjuvant surgical procedures, especially hysterectomy and pulmonary resection for chemotherapy-resistant disease as well as procedures to control hemorrhage, are important components in the management of high-risk gestational trophoblastic neoplasia. Twenty-four (48%) of 50 patients with high-risk gestational trophoblastic neoplasia in this series underwent surgical procedures, and 21 (87.5%) were cured. PMID- 17086806 TI - Changing trends in gestational trophoblastic disease. AB - OBJECTIVE: The New Mexico Tumor Registry (NMTR) and Surveillance, Epidemiology and End Results (SEER) registries were utilized to determine (30+)-year trends in gestational trophoblastic disease and choriocarcinoma. STUDY DESIGN: Age-adjusted incidence rates of gestational trophoblastic disease per 100,000 woman-years (1973-2003) and ratios per live births and pregnancies were calculated using data abstracted from the NMTR and state vital records. SEER data (1973-2002) were used to calculate age-adjusted incidence rates, estimated annual percentage change (EAPC) and relative survival rates for choriocarcinoma. RESULTS: In New Mexico there were 1,153 cases affecting 377 non-Hispanic whites, 504 Hispanics and 241 American Indians, with respective incidence rates of 3.494, 5.150 and 9.991 (p < 0.0001). American Indian incidence rates decreased 53.3%, from 13.34 (1988-1992) to 6.23 (1998-2002). Within SEER (1973-2002), there were 504 gestational choriocarcinomas. The 30-year incidence rate was 0.132 and decreased by 37.7% (EAPC, -2.1% per year; p=0.0001)-by 40.1% for whites, 55.9% for blacks and 62.1% for others. However, over the previous 10 years, rates among blacks (0.097 vs. 0.259, p = 0.01) and for distant disease (0.044 vs. 0.071, p = 0.046) increased. CONCLUSION: Disparities in incidence rates by race/ethnicity in New Mexico are decreasing. An increase in rates among blacks and distant disease diagnosis may be the consequence of fewer regional trophoblastic centers in the United States. PMID- 17086807 TI - Management of gestational trophoblastic neoplasia with metastasis to the central nervous system: A 12-year review at the Philippine General Hospital. AB - OBJECTIVE: To evaluate the clinical characteristics, treatment modalities and outcomes of patients with intracranial metastases resulting from gestational trophoblastic neoplasia (GTN). STUDY DESIGN: A retrospective study was done of patients with brain metastases due to GTN admitted to the Trophoblastic Diseases Section, Department of Obstetrics and Gynecology, Philippine General Hospital, from January 1992 to December 2004. Systemic chemotherapy in the form of methotrexate, etoposide, actinomycin D, cyclophosphamide and vincristine was the treatment of choice. Concomitant whole brain irradiation at a dose of 2,000-3,000 cGy (in 10 fractions of 200-300 cGy) was also given. RESULTS: During 1992-2004, 30 patients with stage IV GTN (brain metastases) were diagnosed based on history, physical examination and computed tomography of the brain at the Philippine General Hospital. Of the 30 patients, 17 (56.7%) belonged to the "early" group (having central nervous system [CNS] symptoms on presentation), while 13 (43.3%) were in the "late" group (individuals who developed lesions during chemotherapy or who had relapsed after initial complete or partial remission). Headache was the most common neurologic symptom. Thirteen received etoposide, methotrexate, actinomycin D with cisplatin and etoposide, 5 received EMACE, 6 received methotrexate actinomycin-D and cyclophosphamide and 1 received methotrexate, etoposide, actinomycin-D. Of the 30 patients, 14 (46.7%) received concurrent whole brain irradiation. Eight (27%) patients responded to treatment and were considered in remission; remission was achieved in 6 of 17 (35%) in the early group and 2 of 13 (15%) in the late group. The mean survival time for the early CNS group was 7.3 months; it was 8.3 months for the late CNS group. CONCLUSION: Intracranial metastasis in GTN is a curable disease that carries compromised survival because of difficulty in implementing the treatment regimen, patient noncompliance and late diagnosis. PMID- 17086808 TI - The need for an hCG assay that appropriately detects trophoblastic disease and other hCG-producing cancers. AB - This report was conceived at the 13th World Congress of Gestational Trophoblastic Disease after multiple presentations indicated widespread discrepancies in human chorionic gonadotropin (hCG) use and results. There appears to be a need for a discussion to describe the advantages and limitations of commonly used hCG tests in the management of gestational trophoblastic disease, and to monitor testicular, germ cell and other hCG-producing malignancies. In most countries hCG tests are certified only for pregnancy testing. Use in managing gestational trophoblastic diseases and other malignancies is considered an "off-label" use. Tests are not optimized or calibrated for these applications, and their use and the results therefore have to be considered experimental. Widespread variations in results occur and may lead to needless or inappropriate therapy. It therefore seems important for laboratory directors and treating physicians to familiarize themselves with which hCG test their laboratory is using and, if necessary, to contract an external laboratory for measuring hCG in the management of gestational trophoblastic disease and cancer. PMID- 17086809 TI - Management of gestational trophoblastic disease and other cases with low serum levels of human chorionic gonadotropin. AB - Recent publications show that the measurements of particular human chorionic gonadotropin (hCG) variants are extremely beneficial in the diagnosis, monitoring and treatment of gestational trophoblastic disease (GTD). Here we review the possible sources of hCG and the use of commercial tests in the optimal management of GTD, quiescent GTD,false positive hCG results, placental site trophoblastic tumor (PSTT) detection, nontrophoblastic neoplasms and pituitary hCG. Hyperglycosylated hCG (hCG-H) measurements are ideal for discriminating active (invasive) from inactive (quiescent or benign) disease. hCG-H testing is also more sensitive than regular hCG in detecting recurrent or persistent disease. After excluding false positive hCG results, and in the absence of any radiographic evidence of tumor, hCG-H should be measured before starting chemotherapy or surgery in women presenting with low hCG (<1,000 mIU/mL) with or without a history of GTD. The hCG free beta assay is an invaluable test in discriminating PSTT from other GTDs, thereby aiding the determination of appropriate treatment options. PMID- 17086810 TI - Postpartum choriocarcinoma presentation, management and survival. AB - OBJECTIVE: To determine the clinical presentation and outcome of postpartum choriocarcinoma. STUDY DESIGN: Case note review of patients with choriocarcinoma treated at the Sheffield Trophoblastic Disease Centre. RESULTS: Thirty-five patients were identified between 1977 and 2005. Mean age was 27 years (range, 21 37). Thirty-three patients complained of persistent postpartum hemorrhage, and in 3 cases there were other symptoms. Two patients presented with nongynecologic symptoms. Mean time until diagnosis was 7 weeks postpartum (range, 0-60), with a mean delay from onset of symptoms to treatment of 7 weeks (maximum, 19). Twenty patients had metastatic disease, but this did not correlate with delay in diagnosis. The mean International Federation of Gynecology and Obstetrics score was 10. Multidrug regimens were used in most patients; however, 8 low-risk patients had a complete response with methotrexate alone. The mean survival was 7.8 years (range, 1-21). Two patients died from disease. CONCLUSION: Postpartum choriocarcinoma presents mainly with vaginal bleeding, and there is often a delay in diagnosis despite being under the care of gynecologists. In the small numbers that present with nongynecologic symptoms there is a rapid awareness of the possibility of gestational trophoblastic neoplasia; nevertheless, the outcome may be fatal, especially in the presence of symptomatic brain metastases. PMID- 17086811 TI - Twin mole and viable fetus: The case for misdiagnosis. AB - OBJECTIVE: To report the Sheffield Trophoblast Centre experience of twin molar gestations and review this in the light of international experience. CASE: Thirty patients with possible twin molar gestations were registered from 1986 to 2004 (during which period 7,200 cases of mole were seen). The accuracy of suspected clinical and histologic diagnoses was investigated. RESULTS: In 10 cases twin mole/fetus had been suspected clinically but not confirmed when products of conception were examined. In 3 of these cases the pregnancy had been therapeutically terminated because of clinical (ultrasound) suspicion of coexisting molar pregnancy. In the 19 cases where twin mole/fetus was suspected, central histopathology review was possible in 14 cases. Only 7 were confirmed. In 2 further cases twin molar gestation was diagnosed on specimens referred for central review as partial mole singleton pregnancies. For confirmed cases the pregnancy outcome was term delivery in 5 cases and miscarriage in 4. CONCLUSION: Clinical and histopathologic diagnosis of twin molar pregnancies is inaccurate in many suspected cases; therefore, a second (expert) opinion should be sought. When the diagnosis is accurate, maternal and fetal complications are common. However, in suspected cases the pregnancy may be allowed to proceed, with caution, if the mother wishes. PMID- 17086812 TI - Relapsed gestational trophoblastic neoplasia: A 20-year experience. AB - OBJECTIVE: To review relapsed gestational trophoblastic neoplasia (GTN). STUDY DESIGN: Patients who had relapsed GTN between 1978 and 2001 at Queen Mary Hospital were included in the study. Records were reviewed and data analyzed regarding treatment, follow-up and survival. RESULTS: Eighteen patients with relapsed GTN were identified. Patients' ages ranged from 21 to 56 years, with a median of 34. Eight were classified as low risk, 1 as medium risk and 9 as high risk at the time of diagnosis. Seven, 3 and 8 patients were treated with single-, dual- and multiple-agent chemotherapy, respectively. The median interval between remission and relapse was 6.5 months (range, 1-132). The time interval to relapse did not correlate with patient mortality (Mann-Whitney U test, p = 0.873). Four patients died of the disease, and all of them were classified and treated as low risk at the time of diagnosis. Three were lost to follow-up at some point. The remaining patient had relapsed choriocarcinoma and developed progressive disease despite intensive multiple-modality treatment. The overall survival rate for relapsed GTN was 77.8%. CONCLUSION: Patients with relapsed GTN are salvageable. Failure of treatment seems attributable to patients who defaulted treatment or follow-up and presented late with massive disease. PMID- 17086813 TI - Management of postmolar gestational trophoblastic disease with methotrexate and folinic acid: 15 years of experience. AB - OBJECTIVE: To study the incidence of postmolar gestational trophoblastic disease (GTD) following hydatidiform mole and to evaluate the effectiveness of single agent chemotherapy using methotrexate with folinic acid rescue. STUDY DESIGN: A prospective study of all cases of hydatidiform mole diagnosed and treated in the department of obstetrics and gynecology, Medical College, Calicut, India, was started in June 1990 to determine the incidence of postmolar GTD and the effectiveness of single-agent chemotherapy with methotrexate and folinic acid in postmolar nonmetastatic GTD. RESULTS: For the 15-year period from June 1990 to May 2005, 1,569 cases of hydatidiform mole were diagnosed and managed at our institution. The incidence of postmolar GTD among 1,569 cases of hydatidiform mole was 20.4%. Of the 321 cases of postmolar GTD diagnosed, 284 patients (88.5%) achieved complete remission with the methotrexate/folinic acid regimen. Fourteen multiparous patients (4.4%) underwent hysterectomy with methotrexate/folinic acid and achieved remission. Thus, 92.9% of patients with postmolar GTD had complete remission with the methotrexate/folinic acid regimen. The rest of the cases required multiagent therapy. CONCLUSION: Regular follow-up of patients after evacuation of hydatidiform mole will detect cases of postmolar GTD at an early stage. Single-agent chemotherapy with methotrexate was effective in 92.9% of our cases. PMID- 17086814 TI - 25 years' experience in the treatment of gestational trophoblastic neoplasia in Hungary. AB - OBJECTIVE: To review our clinical experience in the treatment of gestational trophoblastic neoplasia (GTN) over the past 25 years in our national trophoblastic disease center. STUDY DESIGN: Between January 1, 1977, and December 31, 2001, we treated 355 patients with GTN. The patients were between 14 and 53 years of age, with an average of 28.3. Primary chemotherapy was selected based on the patient's stage of gestational trophoblastic tumor (GTT) and prognostic score. RESULTS: We found metastases in 49.3% (175 of 355) of our patients. Of 173 patients, 162 (93.2%) achieved remission as a result of methotrexate therapy. In 11 patients (6.8%) complete remission was achieved by combination chemotherapy, in some cases assisted by operation. Of 68 patients, 63 (92.6%) achieved remission as a result of actinomycin D therapy, and 5 (7.4%) achieved complete remission by combination chemotherapy. Chemotherapy, surgical intervention or other supplementary treatments resulted in 100% successful therapy in cases of nonmetastatic and low-risk metastatic disease. CONCLUSION: According to our experience, methotrexate/folinic acid or actinomycin D should be the primary treatment in patients with nonmetastatic or low-risk metastatic GTN. Patients with resistance to single-agent chemotherapy regularly achieve remission with combination chemotherapy. PMID- 17086815 TI - Trophoblastic reminiscences. AB - Development of the radioimmunoassay in the 1950s and early '60s largely eliminated early problems with human chorionic gonadotropin and permitted the U.K. to offer a national service for gestational trophoblastic disease (GTD) patients. In 1973 a voluntary registration scheme for patients with hydatidiform mole (HM) opened at 3 U.K. locations. The Charing Cross Centre has followed > 35,000 women with HM, and 2,500 have undergone treatment for various forms of GTD. All treated patients are followed indefinitely and the data computerized. Disasters have occurred in 1 country from misinterpretation of erroneous hCG assays. In terms of experience and data collection, the advantages of a specialized service are overwhelming. This society's main thrust should be to ensure that women with GTD in all countries benefit from specialized management. PMID- 17086816 TI - Presidential address. XIIIth World Congress on Gestational Trophoblastic Disease, Hong Kong, October 23-26, 2005. PMID- 17086817 TI - Purchasing alliance feels the power. Interview by Bob Kehoe. PMID- 17086818 TI - Stay alert! Tracking recalls. PMID- 17086819 TI - Build a better supply chain with metrics that work. PMID- 17086820 TI - Raise awareness. PMID- 17086821 TI - EtO system improves turnaround time. PMID- 17086822 TI - Role changes create new opportunities and partnerships. PMID- 17086823 TI - Nurses rave and rant about IT. PMID- 17086824 TI - Spinal implant pricing model at work. PMID- 17086825 TI - Ahoy, MLTs. PMID- 17086826 TI - Emerging technologies for emerging infections. PMID- 17086827 TI - The impact of STIs. PMID- 17086828 TI - Generations in the workplace: finding common ground. PMID- 17086829 TI - "Diabesity" fills lab day agenda: state learning forum serves as a national model. PMID- 17086830 TI - The interesting provisions of FMLA. PMID- 17086831 TI - Genetic testing: current and future trends. PMID- 17086832 TI - The outlook for CMS competitive bidding: Part II. PMID- 17086833 TI - How recovery happens. PMID- 17086834 TI - Honoring John Fryer's legacy. PMID- 17086835 TI - Finding a place for sex offenders. PMID- 17086836 TI - Working to improve patient care. PMID- 17086837 TI - Optimizing outsourcing. PMID- 17086838 TI - Confronting barriers. PMID- 17086839 TI - The heart-mind connection. AB - A clear connection exists between mental health and heart disease, which requires assessment, collaboration with other professionals, and behavioral health interventions. Psychosocial factors such as stress, anger, anxiety, and depression promote heart disease. Alternately, the use of mood-enhancing or stress-reducing techniques--such as optimism, meditation, counseling, exercise, and imagery--as well as a lifestyle behavioral change approach can have a positive effect on reducing the incidence of heart disease. Behavioral health treatment can improve medical conditions and quality of life. PMID- 17086840 TI - Lawsuits challenge nursing home placements. PMID- 17086841 TI - An image of its former self. PMID- 17086842 TI - At any time, help is on the line. PMID- 17086843 TI - Treating patients with ADHD and coexisting conditions. PMID- 17086844 TI - The aesthetics of healing. PMID- 17086845 TI - Security realities for behavioral health. PMID- 17086846 TI - Saving lives and restoring hope. PMID- 17086847 TI - A wider role for EAPs. PMID- 17086848 TI - Staying alert. PMID- 17086849 TI - We're not ready for P4P. PMID- 17086850 TI - Be warned of seismic activity ahead. PMID- 17086851 TI - Tuberculosis of the temporomandibular joint: low prevalence or missed diagnosis? PMID- 17086852 TI - The immediate effect of changing mandibular position on the EMG activity of the masseter, temporalis, sternocleidomastoid, and trapezius muscles. AB - This study investigated the immediate effect of changing mandibular position on the electromyographic (EMG) activity of the masseter (MS), temporalis (TM), sternocleidomastoid (SCM) and trapezius (TR) muscles. Thirty-three (33) asymptomatic subjects (16 males and 17 females), ages 23 to 52 were selected. Surface EMG recordings were obtained for all muscles bilaterally with the mandible in a relaxed open position (relaxed) and during maximal voluntary clenching (fullbite) for the following: a non-repositioning appliance (NONREPOS) and repositioning appliance (REPOS). REPOS significantly reduced EMG activity of all muscles bilaterally during fullbite. During relaxation, reduction in EMG activity was only found for TR bilaterally. NONREPOS decreased the EMG activity bilaterally for TM and TR and unilaterally (left) for MS and SCM during fullbite. During relaxation, NONREPOS decreased muscle activity bilaterally for TR and SCM. A unilateral reduction was found for TM (right). These findings suggest that immediate alterations in mandibular position affect the cranio-cervical system. Both mandibular positions tested lowered the EMG activity of masticatory and cervical muscles in the relaxed and fullbite positions. The trapezius muscle was the most responsive to alterations in mandibular position. PMID- 17086853 TI - Occlusal characteristics in subjects with facial pain compared to a pain-free control group. AB - Facial pain has been considered a common symptom of temporomandibular disorders (TMD) with a multifactorial etiology. There is controversy regarding the role of occlusion in the background of facial pain and TMD. The aim of the study was to compare the occlusal relationships with two definite measurements in subjects with and without facial pain, in a population-based sample of young adults. The study is part of the Northern Finland 1966 Birth Cohort Project. A subsample of the cohort was formed based on a questionnaire and consisted of 104 subjects, including 52 subjects with facial pain and 52 non-pain controls. Analyses of the dental occlusion of the subjects were performed in gypsum casts by following two methods: 1. the Peer assessment rating (PAR), according to Richmond, et al. (Method 1), and 2. the bilateral canine relationship and the dental midline measurement by the method presented by Pirttiniemi, et al. (Method 2). Method 2 showed higher sensitivity in detecting sagittal occlusal discrepancies than Method 1. Assessment of the intermaxillary relationships in terms of the canine relation showed the lower canine to be more mesially located in the facial pain group, compared to the controls, measured by Method 2. It can be concluded that differences in occlusal sagittal relationships, especially mesial canine relation, seem to correlate with facial pain symptoms at population level. PMID- 17086854 TI - Psychophysiological definitions of clenching. AB - This study tested the hypothesis that individuals show considerable variability in EMG activity produced by the masticatory muscles when they are instructed to clench than when they are instructed to make minimal or maximal contact. Twenty individuals without temporomandibular disorder (TMD) pain participated in a biofeedback-training task to establish a relaxed baseline. They were instructed to clench their teeth according to their personal definition of the term, while EMG data were collected. This process was repeated two more times, followed by similar instructions to make minimal and maximal contact between the teeth. Results showed that individual subjects were very consistent in their behavioral definition of clenching and that the subjects taken as a whole showed markedly greater variability. The precise behavioral meaning of clenching varies across individuals. The failure to account for these individual differences may explain in part reported discrepancies on the role of parafunctions in TMD. PMID- 17086855 TI - A preliminary protocol for multi-professional centers for the determination of signs and symptoms of temporomandibular disorders. AB - The objective of the present study was to test a protocol for the quantification of the frequency and severity of signs and symptoms of temporomandibular disorders (TMD) according to patient perception during two phases of investigation. The protocol was developed based on the signs and symptoms most frequently reported in the literature and on the circumstances in which they produce discomfort. Eighty-four patients diagnosed with TMD by functional examination of the masticatory system responded to the protocol questions and indicated the severity of signs and symptoms using an eleven point numerical scale (Phase 1). Forty-two patients were fitted with an occlusal splint (treated group) and the remaining participants did not use a splint (control group). The protocol questions were asked after 50 days of treatment (Phase 2). Based on the results of nonparametric statistical analysis, the incidence of signs and symptoms was high in Phase 1 and significant, with no difference between the groups, whereas the treated and control groups differed in Phase 2. A comparison between Phases 1 and 2 showed that only the treated group presented a reduction in the severity of signs and symptoms. The study showed that using this protocol, it is possible to define the frequency and severity of symptoms as well as the effect of the treatment. The advantage of this protocol is that it would complement the data obtained using clinical examination with information provided by the patient in a measurable manner. PMID- 17086856 TI - A 5-year follow-up of temporomandibular disorder treatment emphasizing condylar asymmetry. AB - The purpose of this study was to investigate the long-term effect of temporomandibular disorder (TMD) treatment giving priority to oral stability, i.e. creating stability for the mandible in the individual retruded position of the mandible (RPM), focusing condylar asymmetry. Twenty-nine (29) patients treated for TMD participated in the study. A standardized TMD examination was executed originally and an identical examination was performed at the follow-up in a private TMD practice. In general, the patients showed a significant increase of their maximum active mouth opening capacity (p < 0.01) and a significant decrease of dynamic and static pain at the performed tests (p < 0.001). Patients initially suffering facial pain showed a significant reduction of their pain with treatment (p < 0.01). The patients with condylar asymmetry >10%, and who underwent treatment aiming at permanent oral stability, showed the greatest improvement. PMID- 17086857 TI - Quantitative analysis of the variability of unilateral chewing movements in young adults. AB - Kinesiography can be used as a diagnostic tool in a dental clinic context. In the current study, a kinesiograph was used to detect and record the three-dimensional motion of the mandibular mid-incisor point during unilateral chewing as a function of time. The aim of the study was to quantify the within-subject short term reproducibility of the kinesiographic recordings in normal, healthy subjects. Ten seconds of unilateral (right and left) gum chewing were recorded in 20 control subjects using computerized kinesiography. Each subject performed 18 chewing sequences (three repetitions x three sessions x two sides). Chewing cycle duration, volume, standardized depth and width, and the number of reversed cycles were calculated. Intraclass correlation coefficients (two-way random effects analysis of variance with interactions) and paired t-tests were used to compare sessions. For each subject and side, chewing variability was expressed as the coefficient of variation (percentage ratio of standard deviation to mean) of each variable. Mean left and right side mastications were computed over all sessions and subjects. For all the analyzed variables, larger variations between subjects (analysis of variance, p < 0.001) than between sessions were found, with intraclass correlation coefficients ranging between 0.432 (left side cycle duration) and 0.989 (right side standardized width). No systematic errors between the three measurement sessions were found for cycle volume and shape (paired t, p > 0.05). The highest between subjects/ between sessions variance ratios (up to 223.28) were found for cycle duration and shape. In all subjects, chewing cycle volume was very variable, with mean coefficients of variation up to 47% (left side in females). Cycle duration and standardized depth and width were more reproducible, with mean coefficients of variation up to 10% (duration), 14% (standardized width), and 18% (standardized depth). The spatial characteristics of gum chewing cycles had a large within-subject variability. The temporal and size-standardized (shape) characteristics were more consistent within subject. The results should allow selection of a set of relatively more consistent variables for the definition of normality and the comparison of patients. PMID- 17086858 TI - Manual physical therapy interventions and exercise for patients with temporomandibular disorders. AB - The purpose of this study was to investigate the outcome of a series of consecutive patients with temporomandibular disorder (TMD) who were treated with manual physical therapy interventions and exercise. Consecutive patients with the clinical presentation of TMD completed several self-report measures and underwent a standardized historical and physical examination. Following the examination, patients received a multimodal treatment approach incorporating manual physical therapy and exercise. All self-report questionnaires were completed at a 2-week follow-up. Paired t-tests were performed between the baseline and 2-week follow up scores. The mean TMD Disability Index scores were 32.1% (15.4%) at baseline and 18.3% (12.5%) at the 2-week follow-up, representing an improvement of 13.9% (CI: 8.2%, 19.5%) (p < 0.05). Patient Specific Functional Scale (PSFS) scores improved 3.1 points (CI: 2.3, 3.9) (p < 0.05). These results suggest that patients with TMD who are treated with a rehabilitation program including manual physical therapy interventions plus exercise, with or without iontophoresis with dexamethasone, can demonstrate clinically meaningful improvements in disability and overall perceived change in a relatively short period of time. PMID- 17086859 TI - The use of a bruxchecker in the evaluation of different grinding patterns during sleep bruxism. AB - A variety of problems involving the masticatory system can be partially attributed to parafunctional habits such as bruxism. These include occlusal trauma, abfractions, tooth migration, as well as temporomandibular dysfunction. Since bruxism is considered a contributing factor to the above-mentioned dental problems, it is essential to consider parafunctional habits in the diagnosis and treatment planning before doing any occlusal reconstruction. However, the problem lies in the lack or absence of a simple device or gauge useful to be able to diagnose and evaluate the occlusal schemes in the patient's grinding pattern. In this study, we have developed a very simple device (BruxChecker) for evaluating the grinding patterns in sleep bruxism. Using the BruxChecker, it was possible to visualize real or actual interferences during sleep bruxism. Therefore, examination of the grinding pattern using this device is necessary and crucial for making the proper treatment plan for each patient. PMID- 17086860 TI - SUNCT syndrome associated with temporomandibular disorders: a case report. AB - This case report relates the association between a rare neuralgiform syndrome (SUNCT) and masticatory myofascial pain (TMD); two different diseases with different diagnosis criteria and treatments. SUNCT syndrome was treated with a balloon microcompression procedure of the trigeminal ganglion, and the myofascial pain with injections and physical therapy. The patient was without pain at a twelve-month follow-up evaluation. PMID- 17086861 TI - Thermal ablation therapy for hepatocellular carcinoma: comparison between radiofrequency ablation and percutaneous microwave coagulation therapy. AB - BACKGROUND/AIMS: To evaluate the initial therapeutic effect and safety of radiofrequency ablation (RFA) and percutaneous microwave coagulation therapy (PMCT) for the treatment of small hepatocellular carcinomas (HCCs). METHODOLOGY: Forty-eight patients with HCC were treated by RFA and 70 patients with HCC were treated by PMCT. These procedures were repeated until complete tumor necrosis was confirmed by contrast CT scanning. The therapeutic and adverse effects were compared between the two procedures. RESULTS: 1) The number of treatment sessions was significantly lower in RFA patients, and the necrotic area was significantly larger. 2) The local recurrence rate was significantly lower after RFA than after PMCT, while the ectopic recurrence rate showed no significant difference between the two procedures. 3) The survival rate was significantly higher after RFA compared with PMCT. 4) The incidence of pain and fever after treatment was significantly higher in PMCT patients. Occurrence of bile duct injury, pleural effusion and ascites were also significantly more common in PMCT patients. CONCLUSIONS: RFA is more useful for the treatment of small HCCs compared with PMCT because it is minimally invasive and achieves a low local recurrence rate, a high survival rate, and extensive necrosis after only a few treatment sessions. PMID- 17086862 TI - Cholecystectomy improves long-term success after endoscopic treatment of CBD stones. AB - BACKGROUND/AIMS: The aim was to study prospectively primary endoscopic treatment of CBD stones and further the long-term need for renewed gallstone disease interventions, defined as short- and long-term outcome. METHODOLOGY: Seven years prospective follow-up of 101 consecutive patients with CBD stones who underwent endoscopic treatment with the intent of primarily achieving duct clearance. RESULTS: Many patients underwent several endoscopy sessions before stone clearance was completed in 83%. Eleven patients were treated surgically, 2 patients received a permanent stent, and the remaining 3 became stone free with other means. Complications occurred in 47 patients. During follow-up, 31 patients were readmitted for gallstone disease and 15 of these had recurrent CBD stones. Ten percent (8/78) of patients with the gallbladder in situ had acute cholecystitis during follow-up and late cholecystectomy was carried out in 22%. Risk factors for new gallstone disease were an in situ gallbladder containing stones and previous episodes of CBD stones. CONCLUSIONS: A goal of complete CBD stone clearance with ERC and ES proved to be relatively resource consuming. Subsequent cholecystectomy after duct clearance for CBD should be advised when the gallbladder lodges gallstones, especially in younger patients. Recurrent CBD stones were not influenced by cholecystectomy. PMID- 17086863 TI - Radical surgery for gallbladder carcinoma: possibilities of survival. AB - BACKGROUND/AIMS: An aggressive surgical approach in the management of gallbladder cancer (GBC) has improved survival significantly in recent years. The aim of this retrospective study is to evaluate the long-term results of surgical treatment of GBC reassessed following the TNM staging system of the AJCC-2002. METHODOLOGY: The present series considers 118 patients with GBC treated between 1982 and 2003. Seventy-four cases (63%) were females and 44 (37%) males; overall age was 63 years (range 38-91). RESULTS: Among the 118 patients with GBC, 35 (36%) underwent radical surgery: 3 pTNM IA [3 cholecystectomy (CT)], 10 IB [3CT, 3 CT + hepatic resection (HR), 4 HR], 3 IIA (3 HR), 7 IIB (3 CT+HR+ bile duct resection (BDR), 3 HR, 1 hepatopancreatoduodenectomy +CT), 10 III [4 CT+HR, 5 extensive HR (eHR), 1 HR+right colectomy+BDR+total gastrectomy], 2 IV (2 eHR). Overall 1-, 3-, 5-year survival was 67%, 46%, 34% respectively for stage IA-IB; 63%,12%, 12% for IIA IIB; 50%, 30%, 30% for III-IV (p=ns); in particular, 1-, 3-, 5-year survival was 100%, 100%, 100% for T1a; 50%, 50%, 50% for T1b; 70%, 46%, 35% for T2; 50%, 12%, 12% for T3; 54%, 32%, 32% for T4 (p=ns); 1-, 3-, 5-year survival for patients without lymph node involvement was 58%, 44%, 37% and 60%, 15%, 15% for patients with lymph node metastases (p=ns), respectively. CONCLUSIONS: CT seems to be sufficient in T1a GBC patients but inadequate in T1b (stage IA), which requires a more aggressive approach. In stage IIB, III and IV, the presence of lymph-node metastasis is not a contraindication to aggressive surgery. PMID- 17086864 TI - Decreased interstitial cells of Cajal in the neorectum after anterior resection of the rectum. AB - BACKGROUND/AIMS: Patients who have undergone anterior resection for rectal carcinoma often complain of anorectal and defecatory dysfunction postoperatively. The aim of this study was to examine the expression of interstitial cells of Cajal (ICCs) in the sigmoid colon used for constructing the neorectum after anterior resection of the rectum. METHODOLOGY: As the neorectum group, we assessed 12 patients with local and anastomotic recurrence or new neoplasm in the neorectum after anterior resection of the rectum. The control group consisted of 16 patients who underwent sigmoid colon resection for sigmoid colon carcinoma. All resected specimens were investigated with immunohistochemical staining, using c-kit antibody for ICCs. The correlation between the number of ICCs and defecatory symptoms was assessed for the neorectum. RESULTS: The total number of ICCs significantly decreased in the neorectum group as compared to the control group. In particular, a significant difference was noted between the two groups as to the number of ICCs found between the layers of the myenteric plexus in histological studies, as well as in the circular and longitudinal muscles. There was no correlation between the number of ICCs and the time interval from the initial anterior resection to the resection of the neorectum, nor was there any relationship between the number of ICCs and defecatory symptoms. CONCLUSIONS: The expression of ICCs in the neorectum was reduced in the early stages after anterior resection of the rectum. Expression of ICCs in the neorectum did not recover to preoperative levels over time. PMID- 17086865 TI - Gastrointestinal fiberscopic findings after simple closure for perforated duodenal ulcer in the early postoperative phase: from experience to evidence by evaluation of the healing process. AB - BACKGROUND/AIMS: Today, in light of widespread adoption of H2-RA and PPI, the standard surgical procedure for perforated duodenal ulcer (PDU) is simple closure and/or omental patch (SC). However, the healing process after these techniques has not been fully examined. We have not yet confirmed the propriety of simple suture of the bottom of the ulcer. This technique has been performed based only on experience, and there is insufficient evidence to conclude that this procedure can be definitively considered a safe therapeutic technique for the majority of patients with PDU. The aim of this study is to clarify the macroscopic findings of the healing process after SC for PDU. METHODOLOGY: Thirteen patients with PDU who were treated with SC underwent postoperative gastroduodenal fiberscopy (GF) at the 7th-16th postoperative day and the healing process was monitored under sufficient informed consent. Patients with severe preoperative disease were excluded from the study. Healing condition of the ulcer and stitches, deformity, and stenosis were evaluated by postoperative endoscopy. Possible adverse effects that were evaluated included: perforation, rise in fever, worsening of inflammation on laboratory data, gastrointestinal symptoms such as abdominal pain, sense of fullness, and vomiting. The indications for SC were as follows: PDU with 1) no stenosis and 2) no prominent ulcer ridge. The surgical technique was as follows: 1) interrupted simple closure with no trimming and debridement of wound (4-5 stitches) with absorbable monofilament suture, and/or 2) omental patch, 3) administration of H2-RA (or PPI) just after operation, and 4) oral feeding 4-5 days after operation independent of postoperative GF. RESULTS: GF findings in 2 patients showed active and healing stage, in whom surgical technique was thought to be insufficient; the ulcer had been large and included a descending portion, or a small perforation had occurred in the large ulcer bottom (the distance between the stitches and the edge of the ulcer was insufficient). In the other 11 patients, GF findings showed scar phase. There was no morbidity related to endoscopic procedure. CONCLUSIONS: Sutured PDU with SC will be in the scar phase in 1 or 2 weeks. Postoperative GF 1 week after SC for PDU is thought to be a safe examination. This study is a primitive study of a small group, and more cases that can adequately show the frequency of complications and indicate the overall safety of the procedure are needed. PMID- 17086866 TI - Carbohydrate malabsorption in clinical routine: a prospective observational study. AB - BACKGROUND/AIMS: Nonspecific abdominal symptoms are a serious problem throughout the world. Among the multitude of differential diagnoses in carbohydrate malabsorption, only incomplete absorption of lactose is mentioned, while malabsorption of fructose and sorbitol--which occurs much more often, at least in the Western world--is usually not included. METHODOLOGY: During a 6-month period, all patients (n=90; 33 males, median age 45 years, range 10-81; 57 females, median age 47 years, range 15-71) who consecutively presented for H2 exhalation tests were evaluated. In addition to the test results, data were obtained from the referring physicians and from the family doctors responsible for the patients' long-term treatment regarding the role of the test results in the treatment of the patients. Finally, the patients were also asked whether any improvement in their symptoms had followed from the test results. RESULTS: Lactulose tests were normal in only 63% of the patients. As with the other sugars, at least one form of malabsorption was detected in 47 patients (52%). The malabsorption rate was 34% after lactose, 61% after fructose, and 91% after the intake of sorbitol. The referring physicians evaluated the test results as having been important in 52% of the patients, while the family doctors considered that there was some benefit for the patients in 77% of the cases. The patients themselves reported an improvement in 75% of cases. CONCLUSIONS: These data again show that carbohydrate malabsorption is an important differential diagnosis in patients with nonspecific abdominal complaints. However, the data also make it clear that caution is advisable both in establishing the indication for the tests and in interpreting the results. Despite this, carbohydrate malabsorption appears to be an underestimated problem in a considerable number of patients. PMID- 17086867 TI - Usefulness of serum hepatic fibrosis markers in the diagnosis of nonalcoholic steatohepatitis (NASH). AB - BACKGROUND/AIMS: In the present study, we examined the usefulness of serum hepatic fibrosis markers for the diagnosis of nonalcoholic steatohepatitis (NASH). METHODOLOGY: The subjects were 16 patients with NASH and 9 patients with fatty liver (FL). All were negative for serum HBsAg, HCVAb, antibodies related with autoimmune diseases, alcohol intake, and drug abuse. We measured the biochemical markers for liver function, hepatic fibrosis markers such as type III procollagen N-peptide (PIIIP), type IV collagen (TyIV), hyaluronic acid (HA) and leptin, and compared these data with histological findings of biopsy specimens. In addition, we examined the diagnostic efficiency of fibrosis markers and leptin for NASH using receiver operating characteristic (ROC) curve. Body mass index (BMI), fasting blood sugar, triglyceride, and degree of fat droplets, inflammation, iron deposition and fibrosis were significantly higher in the NASH group compared with the FL group. RESULTS: The diagnostic efficiencies of NASH% (cut-off value) were 68% (100ng/mL) for TyIV, 68% (10ng/mL) for HA, 64% (0.62U/mL) for PIIIP and 56% (8pg/mL) for leptin. CONCLUSIONS: From these results, it is suggested that the serum hepatic fibrosis markers such as TyIV, in addition to liver biopsy, may be useful for the diagnosis of NASH. PMID- 17086868 TI - Risk factors for anastomotic leakage after resection for rectal cancer. AB - BACKGROUND/AIMS: The aim of this study was to identify risk factors for anastomotic leakage in patients that have received a resection for rectal cancer. METHODOLOGY: Between January 1996 and December 2002, 499 patients underwent rectal resection for rectal cancer performed by the same surgeon, and of these 25 patients developed anastomotic leakage. For this case-control study, we selected two age- (not more than 5 years), sex-, and date of operation- (not more than 3 months) matched controls per case from our computerized database. We collected the data on these 75 patients from the prospectively recorded database and by chart review. RESULTS: The level of the anastomosis from the anal verge and operation-related blood transfusion were found to be independently significant anastomotic risk factors by multivariate analysis. The risk of anastomotic leakage was 5.32 times higher for an anastomosis situated less than 5cm from the anal verge (P=0.006; 95% confidence interval 1.608-17.252), and was 3.90 times higher for patients that received an operation-related blood transfusion (P=0.032; 95% confidence interval 1.120-13.207). The 95% confidence interval of the mean level of the anastomosis from the anal verge in the leakage group was 3.5-4.5cm, and the 95% confidence interval for the mean operation-related blood transfusion in the leakage group was 0.2-1.4 packed red blood cells. CONCLUSIONS: We recommend proximal diversion in patients that have an anastomosis just above the anorectal ring (3.5-4.5cm from the anal verge), or received intraoperative transfusion (more than 0.2-1.4 packed red blood cells). PMID- 17086869 TI - Possible impact of autologous blood towards elective open and laparoscopic surgery for colorectal carcinoma. AB - BACKGROUND/AIMS: An alternative approach to the allogeneic transfusion is patient's blood collection before the operation, using or not using the human recombinant erythropoietin. The aim of this study is to evaluate the percentage of people undergoing an elective operation for colorectal carcinoma who could avoid allogeneic blood due to blood predonation. METHODOLOGY: From January 1999 to April 2002, 249 patients (140 males and 109 females) underwent an operation for colorectal cancer. The most important parameter, used to estimate which patients are candidates for autologous blood predonation, is the before-treatment hemoglobin level. The other parameters we considered are the age of the patient, the absence of uncontrolled hypertension or clinically significant pathologies (except colorectal cancer), and the iron shortage. DESIGN: Retrospective cohort study. RESULTS: 36.5% of the patients satisfied every criterion to begin autologous predonation without using the human recombinant erythropoietin; instead, 23.7% of patients needed the administration of the human recombinant erythropoietin and additional iron. The valuation of the transfusional prospects, based on sex and cancer location is very interesting. CONCLUSIONS: Autologous blood predeposit in preoperational time turns out to be a method of increasing importance, thanks to the human recombinant erythropoietin. PMID- 17086870 TI - In vitro thermochemotherapy of colon cancer cell lines with irinotecan alone and combined with mitomycin C. AB - BACKGROUND/AIMS: Irinotecan (CPT-11) is a new drug of the camptothecin family which has shown significant activity in the treatment of metastatic colorectal cancer. Hyperthermia has been shown to enhance the cytotoxic effect of some anticancer drugs and has been combined with intraperitoneal chemotherapy for the treatment of colorectal peritoneal carcinomatosis. The purpose of this study was to evaluate the cytotoxic effect of CPT-11 alone and in combination with mitomycin C (MMC) and hyperthermia on three colorectal cancer cell lines: CACO-2, HT-29, and DHD/K12/TRb (PROb). METHODOLOGY: The cytotoxic effect of CPT-11 was tested at seven different concentrations (from 2.5 to 160microg/mL) for each type of cell line at 37, 39, 42.5 and 44 degrees C. Combined cytotoxic effect of MMC with CPT-11 was tested at 37 and 42.5 degrees C. RESULTS: The cytotoxic effect of CPT-11 alone increased with concentration (p<0.001) and with increasing temperature (p<0.001) at concentration above 5microg/mL in all three cell lines. CPT-11 (20microg/mL) significantly increased the cytotoxicity of MMC (8microg/mL) at 42.5 degrees C on the CACO-2 line. The combination of CPT-11 and MMC had at least 92% cytotoxicity on the three cell lines. CONCLUSIONS: The combination of CPT-11 and MMC at 42.5 degrees C had a large spectrum of cytotoxicity in these in vitro models. Our findings support the clinical use of this combination and provide a rationale for the design of a clinical trial using intraperitoneal chemohyperthermia with MMC and CPT-11 to treat colorectal peritoneal carcinomatosis of colorectal origin. PMID- 17086871 TI - Operative and long-term results after one-stage surgery for obstructing colonic cancer. AB - BACKGROUND/AIMS: To analyze retrospectively the operative results and five-year survival after single-stage resection and primary anastomosis for right- or left sided colonic malignant obstruction. METHODOLOGY: From 1994 to 2002, 83 patients with acute obstruction due to primary cancer underwent a one-stage procedure, 36 (43.3%) for cancer of the right and 47 (56.7%) of the left colon. Of the 47 patients with left acute obstruction, 45 were treated with intraoperative colonic on-table lavage for decompression and wash-out before resection and anastomosis. Long-term survival was compared with the outcome of 369 patients with colonic cancer without obstruction 144 (39%) right and 225 (61%) left. RESULTS: Overall operative morbidity and mortality were 25.3% and 6%, respectively. Overall anastomotic leakage rate was 6%. Kaplan-Meier curves showed five-year survival rates poorer for patients operated on for obstructing malignancy than those observed for patients without obstruction (p=0.0001, Log-Rank test), obstructing malignancy was associated with more advanced age (p<0.0005) and staging of the cancer. CONCLUSIONS: One-stage resection and primary anastomosis can be applied to the majority of patients with malignant colonic obstruction and it allows achieving excellent operative results. Obstructing colonic cancer proved to be associated to a worse long-term survival. PMID- 17086872 TI - Intraluminal high dose rate brachytherapy of tumorous esophageal stenoses. AB - BACKGROUND/AIMS: The prognosis of inoperable carcinomas of esophagus is poor, and therapeutic efforts are generally limited to palliation. The aim of this study was to retrospectively evaluate the effectiveness of intraluminal high dose rate brachytherapy in the palliative treatment of tumorous esophageal stenoses. METHODOLOGY: Between February 1997 and July 2002 intraluminal brachytherapy was performed in 19 patients with inoperable esophageal carcinoma (squamous cell carcinoma in 14 cases and adenocarcinoma in 5 cases). All patients had dysphagia at presentation. Brachytherapy was performed using high dose rate afterloading system. RESULTS: Dysphagia was improved in all patients. The mean survival was 248 days. No mechanical complications were observed during introduction of the applicator. CONCLUSIONS: Our experience indicates that intraluminal brachytherapy is an effective and safe method of palliation of dysphagia caused by malignant esophageal stenosis. PMID- 17086873 TI - Chemoradiotherapy with and without esophagectomy for advanced esophageal cancer. AB - BACKGROUND/AIMS: To evaluate the impact of surgery on survival after chemoradiotherapy, we analyzed the long-term outcome of patients with advanced esophageal cancer. METHODOLOGY: Data on 92 consecutive patients with T3 or T4 esophageal cancer who were initially treated by chemoradiotherapy were reviewed retrospectively. Of 82 patients who completed the planned schedule, 35 patients underwent esophagectomy (CRT+E Group) and 47 patients received definitive chemoradiotherapy (CRT Group). RESULTS: A response to chemoradiotherapy was obtained in 71% of all 92 patients. The 1- and 3-year survival rates in the patients with T3M0 were 87 and 44 percent respectively, while these in the patients with T4 and/or M1(Lymph) disease were 47 and 20 percent. Although there was no difference in overall survival between the CRT+E Group and the CRT Group, the survival of responders in the CRT+E Group was significantly higher than that of those in the CRT Group (P=0.0448). The locoregional recurrence rate of responders in the CRT Group was higher than that in the CRT+E Group. Multivariate analysis showed that the independent prognostic factors were response, M(Lymph), and esophagectomy. CONCLUSIONS: Although this study was retrospective and nonrandomized, esophagectomy after chemoradiotherapy might improve the survival of responders for locoregional control. PMID- 17086874 TI - Intraoperative manometry of the lower esophageal sphincter pressure during laparoscopic antireflux surgery with a mechanical calibration--early results. AB - BACKGROUND/AIMS: Persistent postoperative dysphagia diminishes the good effect of laparoscopic anti-reflux surgery. An excessive increase of the intraoperative lower esophageal sphincter pressure (LESp) is supposed to be related to the persistent postoperative dysphagia and its knowledge could lead to the modification of the surgical technique followed by improved clinical outcomes. This study aims to describe the relation between the intraoperative LESp increase and the incidence of postoperative dysphagia and to find whether a combination of intraoperative manometry and mechanical calibration of the wrap is able to decrease the incidence of the persistent postoperative dysphagia. METHODOLOGY: The randomized, prospective, two-branch study included 39 patients suffering from symptoms of gastroesophageal reflux disease. All patients underwent pre- and postoperative manometry, 24-hour pH-metry and laparoscopic anti-reflux surgery. The intraoperative LESp was measured in the study arm only. RESULTS: A higher incidence of persistent postoperative dysphagia was revealed in patients with the intraoperative LESp increase more than 15 mmHg. This complication was not found in patients with the LESp increase under 8 mmHg with no impact on the efficacy of the surgery. The combination of the intraoperative manometry and the mechanical calibration of the wrap seems to bring the benefit only to a small number of the patients. CONCLUSIONS: According to our results, the intraoperative LESp measurement proved to be a useful supplementary method which was easy to perform, and which enables a modification of the surgical technique to decrease the incidence of the persistent postoperative dysphagia. PMID- 17086875 TI - Prognostic factors of curatively resected esophageal carcinoma associated with multiple metastatic lymph nodes. AB - BACKGROUND/AIMS: Prognosis of esophageal carcinoma with multiple metastatic lymph nodes is dismal despite radical operation and adjuvant therapy. We investigated prognostic factors for curatively resected esophageal carcinoma with multiple positive nodes. METHODOLOGY: From January 1983 to December 2002, 343 patients with thoracic esophageal carcinoma underwent an esophagectomy with curative intent. Of these patients, 82 patients were associated with 4 or more histopathologically positive nodes. Of these patients, 59 patients underwent a curative resection. Of these 59 patients, 7 patients who died of postoperative complications during the hospital stay were excluded. Therefore, 52 patients were enrolled in this study. Survival curves were compared after stratifications according to 14 clinicopathologic variables. Independent prognostic factors were detected using a multivariate Cox proportional hazard model. RESULTS: The cumulative 5-year survival rate for the subjects was 10.6%. The factors affecting cumulative survival rate by a univariate analysis were intramural metastasis (absence vs. presence) (p=0.03), and postoperative therapy (performed vs. not performed) (p=0.02). A multivariate analysis detected the performance of postoperative therapy (Hazard Ratio= 0.390, p= 0.002) and the absence of intramural metastasis (Hazard ratio=0.429, p=0.01) as positive prognostic factors. CONCLUSIONS: The positive prognostic factors for esophageal carcinoma with multiple lymph node metastases were the absence of intramural metastasis and the performance of adjuvant therapy. PMID- 17086876 TI - Neoadjuvant therapy for esophageal cancer--what can we accept? AB - BACKGROUND/AIMS: We evaluated neoadjuvant use in managing patients with esophageal carcinoma and its effects on the surgical resection and outcomes. METHODOLOGY: Patients prior to esophageal resection were offered the opportunity to receive a neoadjuvant cytostatic regimen (CDDP + FU, CDDP, or TAX + FU). Retrospective tumor chemoresistance analysis using the MTT test was also performed. RESULTS: Seventy patients were operated from 2001 until May 2004. A total of 55 resections were performed with preoperative neoadjuvant therapy and 15 elected to only undergo surgery without neoadjuvant therapy. No deaths occurred as a result of surgery or neoadjuvant therapy, but complications included fistulas and hemorrhages. CONCLUSIONS: There was no significant difference between the postoperative complications among the neoadjuvant and non neoadjuvant groups. This therapy therefore does not have any influence on the course or results of surgical resection. MTT testing did not demonstrate any particular usefulness in tailoring neoadjuvant therapy. Chemoresistance could only be retrospectively evaluated and the results may be affected after cytostatic therapy. The long-term outcomes have not been evaluated yet due to the short follow-up time in our patient group. PMID- 17086877 TI - Bioelectrical impedance analysis to assess changes in body water compartments after digestive surgery. AB - BACKGROUND/AIMS: Accurate monitoring of fluid balance in patients after surgery is a difficult task. Bioelectrical impedance analysis (BIA) is a safe and noninvasive method to measure extracellular water (ECW) and intracellular water (ICW) by passing a weak alternating current through the body. The purpose of the present study was to evaluate changes in body water compartments after gastroenterological surgery by BIA in relation to patient age, type of operation, postoperative complications and systemic inflammatory response syndrome (SIRS). METHODOLOGY: Ninety-four patients undergoing digestive surgery in our department [laparoscopic cholecystectomy (n=9), gastrectomy (n= 23), colectomy (n=26), hepatectomy (n=29), pancreatoduodenectomy (n=4) and esophagectomy (n=3)] were enrolled in the study. Body fluids were measured by bioelectrical impedance analysis before and after surgery (one hour after operation and on postoperative days 1, 3, 7 and 14). RESULTS: Total body water (TBW) and ICW in all groups were significantly lower than preoperative values on day 14. Day 14 ECW in patients less than 70 years or age without postoperative SIRS or complications was significantly lower than the preoperative value. In contrast, ECW was not significantly different from the preoperative value in patients older than age 70 with postoperative SIRS. Additionally, ECW on day 14 was significantly higher than the preoperative value in patients with postoperative complications. When types of surgery were taken into consideration, day 14 TBW was significantly lower than preoperative value only in patients with gastrectomy and hepatectomy. CONCLUSIONS: Development of postoperative SIRS and complications resulted in an increase of ECW above its preoperative value. BIA is useful for detecting small changes in body composition following gastroenterological surgery, and provides a means for monitoring perioperative water balance. PMID- 17086878 TI - Postoperative host responses in elderly patients after gastrointestinal surgery. AB - BACKGROUND/AIMS: The age-associated dysregulation of hemodynamic, metabolic and immune responses contributes to the high incidence of complications after major abdominal surgery. METHODOLOGY: Ninety-five patients who underwent gastric resection (n=51) and colorectal resection (n=44) were divided according to age into Groups A (n=45, less than 70 years old), B (n=30, 70-79 years) and C (n=20, over 80 years). Flow cytometric analysis of CD4+ lymphocytes for interferon (IFN) gamma and interleukin (IL)-4 production determined the Th1/2 balance. Energy expenditure was measured by indirect calorimetry, and hemodynamics were studied using pulse dye densitometry. RESULTS: Surgical procedures, operating time, blood loss and morbidity did not significantly differ among the three groups. The cardiac index (CI) in group A and B increased significantly over preoperative levels until POD 3, but there were no significant perioperative changes in the CI levels of group C. Resting energy expenditure levels changed similarly to those of CI. The postoperative Th1/2 ratio decreased from young to elderly to very elderly patients, although no differences were significant before surgery. The postoperative percentage of CD4+IFN-gamma +T cells (Th1) in group C decreased significantly despite of no significant changes in that of group A and B. In contrast, the ratio of CD4+IL-4+T cells (Th2) in the all groups significantly increased after surgery. CONCLUSIONS: Host responses in elderly patients after major abdominal surgery were more hyperdynamic and hypermetabolic than those of young patients. Postoperative dysregulation of the Th1/2 balance was also associated with aging. However, host responses appear to significantly differ between elderly and very elderly patients. PMID- 17086879 TI - Results of 404 hepatic resections including 80 repeat hepatectomies for hepatocellular carcinoma. AB - BACKGROUND/AIMS: To evaluate our treatment protocol applied to patients with hepatocellular carcinoma. The protocol consists of the selection criteria for hepatectomy, the use of techniques that minimize intraoperative blood loss, strict follow-up after surgery, and an aggressive surgical approach for intrahepatic recurrence. METHODOLOGY: We conducted a retrospective cohort study that included 337 patients with hepatocellular carcinoma treated between 1990 and 2001. The type of resection was selected according to the serum bilirubin value and the indocyanine green retention rate at 15 minutes. Perioperative data and long-term outcome were examined. RESULTS: We performed 324 initial hepatectomies with an in-hospital mortality rate close to zero. There was one operative death and one hospital death (0.3% each), and the 5-year survival rate for all patients was 53.2%. Eighty repeat liver resections, including 18 third and two fourth, were performed with no mortality, and the 5-year survival rate was 52.9% after the second hepatic resection. The resectability rate for second and third hepatectomies reached 29% and 33% of all patients with isolated liver recurrence, respectively. CONCLUSIONS: Liver resection is a safe and effective treatment modality for hepatocellular carcinoma. Our results are likely attributable to the routine application of our treatment protocol. PMID- 17086880 TI - Relationship between hepatocellular carcinoma and impaired glucose tolerance among Japanese. AB - BACKGROUND/AIMS: Both the incidence of diabetes mellitus (DM) and mortality from Hepatocellular carcinoma (HCC) are increasing in Japan. As the association of overall cancer and HCC with impaired glucose tolerance (IGT) has been studied rarely in the world including Japan, this study assessed their associations using cohort data of Hokkaido, Japan. METHODOLOGY: After getting ethical consent, this study included 908 men and 1,081 women aged 30-77 years during 1977-78 and collected detailed information using the baseline survey. The subjects were followed until 2002 and deaths were recorded using ICD-9. Classifying them into three groups of diabetes status namely DM, IGT, and normal, the relative risk (RR) of mortality was estimated by diabetes status using multivariate Cox model. RESULTS: This study revealed no association between overall cancer and diabetes status. However, the RR of mortality from HCC was about 11 times (HR= 10.8, 95%CI: 1.3-92.5) higher in IGT compared with normal group. DM group also showed higher risk of mortality than normal group. CONCLUSIONS: HCC mortality was significantly high among IGT group. However, as the results of the study were based on small data, further studies with large cohort are needed to address the association of IGT with overall cancer and HCC mortality in Japan. PMID- 17086881 TI - Hepatocyte proliferation and apoptosis in rat liver after liver injury. AB - BACKGROUND/AIMS: In this paper the early phase of proliferate response and apoptosis of hepatocytes after partial liver resection, during reperfusion after ischemia and during sepsis is demonstrated. METHODOLOGY: Experiments were conducted in a rat model with regeneration times of 0.5-24 hours after injury. Proliferation was analyzed by Ki-67 immunohistochemistry and confirmed by double staining with CK18 in FACS. Apoptosis was analyzed by TUNEL technique. RESULTS: Periportal hepatocytes enter the cell cycle already 0.5-2 hours after injury in all three models. This early proliferative response is predominant periportally localized. During reperfusion and during sepsis there was a strict pericentral apoptosis of hepatocytes found. CONCLUSIONS: An early periportal proliferation of hepatocytes is a common reaction of the liver to injury. This proliferation takes place much earlier then the main proliferative response 24-72 h after partial resection. This predominant periportal proliferation together with the pericentral apoptosis fit to the concept of the "streaming liver" in liver regeneration. PMID- 17086882 TI - Reliability and diagnostic use of a test for the search of the hepatitis C virus Ag (AgHCV). AB - BACKGROUND/AIMS: The detection of serum HCV nucleocapsid (core) antigen, besides being a valid alternative, by virtue of its low cost, to direct analysis of the virus in everyday transfusion practice, also aims to be employed in monitoring patients subjected to antiviral therapy. The verification of strict correlation between the two tests is the presupposition for such use. METHODOLOGY: In a group of 112 HCV-positive subjects, we assessed blood transaminases, viremia (by PCR), and the circulating core antigen (by ELISA). RESULTS: Only 16 out of 112 patients were AgHCV-negative, with viremia levels in the 10(2) to 10(4) range; 96 patients were HCV-positive, as indicated both by viremia and by Ag detection (1.9 to 292.4pg/mL). Sensitivity of the ELISA test corresponds to 3.6x10(4) IU/mL of viral load. There is an evident aggregation of results in groups according to antigenemia classes and the corresponding viremia levels: <10pg/mL--10(4) IU/mL (6/96); up to 100pg/mL--10(5) IU/mL (55/96); 100-200pg/mL--10(6) IU/mL (31/96); and more than 200pg/mL--10(7) IU/mL (4/96). CONCLUSIONS: AgHCV is correlated with elevation of ALT and high or medium-high viral loads. It can discriminate between ongoing and previous infection and is suitable for monitoring the pharmacological therapy in the presence of sufficiently high viral loads and for evaluating the onset of medium-long-term relapses. Neither the genotypes nor pharmacological therapy appear to affect the comparison of viremia with antigenemia. Moreover, this analysis is cheaper as compared with molecular techniques. PMID- 17086883 TI - Surgical management for metastatic liver tumors. AB - BACKGROUND/AIMS: This study investigated the results of hepatectomy for multiple liver metastases and repeated hepatectomy for recurrent hepatic metastases. A proposed treatment strategy for liver metastases is discussed. METHODOLOGY: Fifty seven consecutive cases of liver metastases were studied. The metastases originated from colon cancer (24 cases), rectal cancer (11 cases), gastric cancer (14 cases), or gastrointestinal stromal tumors (two cases). The other cases included one each of gastric carcinoid, carcinoma of the papilla of Vater, cystic duct cancer, esophageal cancer, choriocarcinoma and breast cancer. RESULTS: The overall 5-year survival rate for the 57 cases was 45.4%; there was no significant difference between patients with colon cancer (56.3%), rectal cancer (45.5%), or gastric cancer (41.6%). The cumulative 5-year survival rates for synchronous and metachronous metastases were 38.3% and 50.8%, respectively (difference not statistically significant; NS). The survival rates for single and multiple metastases were 56.0% and 31.3% (NS), and those for monolobar and bilobar metastases were 48.5% and 40.9% (NS), respectively. Concerning the operative procedure, the survival rates for partial resection and hemi-hepatectomy were 49.5% and 26.9%, respectively (NS). The survival rates for surgical margins <4mm and >5mm were 45.9% and 45.4%, respectively (NS), and those for single and repeat hepatectomy were 40.5% and 58.2% (NS). Preoperative portal embolization was performed in seven cases because of multiple metastases or a tumor located in a deeper site in the liver. There was no hospital death among the 57 cases. CONCLUSIONS: These results show that hepatectomy may offer longer survival, even in patients with multiple or bilobar metastases. Neither the operative procedure nor the size of the surgical margin had any influence on survival after hepatectomy. The prognosis was improved not only for metastases from colorectal cancer, but also for gastric cancer. An increased survival benefit was obtained by repeat hepatectomy for recurrent hepatic metastases. Preoperative portal embolization extended the indication for hepatectomy and provided postoperative safety. PMID- 17086885 TI - Liver resection for large-size hepatocellular carcinomas in 47 non-cirrhotic patients--no mortality and long-term survival. AB - BACKGROUND/AIMS: Few reports have analyzed short- and long-term outcomes in the subset of patients with hepatocellular carcinoma (HCC) on non-cirrhotic liver. METHODOLOGY: From January 1985 to December 2002, 277 patients underwent liver resection for HCC; in only 47 the liver was normal or showed mild chronic hepatitis at histology. RESULTS: A major hepatectomy (MHR) was accomplished in 37 cases (78.7%) including an extended hepatic resection in 18 (38.3%). In-hospital mortality was nil. The rate of complications was 40.4%. Overall and disease-free survival rates at 5 years were 30.9% and 33.9%. Fifteen patients are actually alive with a median survival of 33.3 months. By multivariate analysis, tumor size > 10cm and presence of satellite nodules were independent predictive factors of 5 year survival; median survival of thirteen patients with HCCs < or = 10cm and without daughter nodules was 60 months. Twenty-six patients had a margin less than 1cm and without cancer involvement; overall and recurrence-free survival rates were comparable to those of the patients with a > 1cm margin. CONCLUSIONS: In the treatment of HCC without cirrhosis, major hepatic resections are often needed. Tumors less than 10cm in size and without satellite nodes are the best candidates for operation. The width of the resection margin is unimportant provided that there is no microscopic infiltration. PMID- 17086884 TI - Split liver transplantation for hepatocellular carcinoma. AB - BACKGROUND/AIMS: Liver transplantation (LTx) is recognized as the treatment of choice for small hepatocellular carcinomas (HCC) in patients with end-stage liver failure. However, because of limited organ availability, not all those who qualify can benefit from full-size LTx. METHODOLOGY: Over a 28-month period, we transplanted 6 deceased donor split liver allografts in stable recipients with HCC and cirrhosis. RESULTS: There were 5 men and one woman with a median age of 60 years. Median waiting time to LTx was 82 days. Extended right split grafts (segments I, IV-VIII) were transplanted in 5 patients. One patient received a right split graft. In situ harvesting was performed in 4 instances and ex situ in the remaining 2. Median cold ischemia time was 10 hours. Primary non-function was not observed. Median intensive care unit stay was 4 days. There were neither vascular nor biliary complications. The postoperative course was uneventful in half of the patients. Two patients were re-operated because of an abscess caused by a liver necrosis in the marginal zone of segment IV. One patient died for reasons unrelated to liver function or to the surgical intervention. The remaining 5 patients are alive after a median follow-up of 20 months. CONCLUSIONS: Deceased donor split LTx constitutes an additional option for patients with HCC and cirrhosis. The potential risks of using "split livers" as well as the potential benefits of transplanting patients unlikely to survive the waiting list period must be evaluated on an individual basis. PMID- 17086886 TI - Postoperative changes in remnant medial segment parenchyma of living donor livers after procurement of left lateral segment graft. AB - BACKGROUND/AIMS: Procurement of left lateral segment (LLS) graft usually resulted in uneventful atrophy of the remnant medial segment parenchyma (RMSP) in living donors, but duct dilatation was often detected in the RMSP. We tried to clarify the postoperative atrophic sequences of the RMSP. METHODOLOGY: We analyzed clinical profiles of 33 LLS graft donors out of 362 living donors from January 2002 to December 2003. LLS grafts were divided into type 1 (classical LLS, n= 18), type 2 (intermediately extended LLS, n=8), and type 3 (extended LLS similar to left lobe without middle hepatic vein trunk, n= 7). RESULTS: On 1-week dynamic computed tomography, early extensive hyperattenuation indicating interlobar arterial collaterals (ILAC) was observed in 4 (22.2%) donors of type 1 and 2 (25%) donors of type 2 LLS grafts. Peak values of serum liver enzymes were not correlated with the amount of RMSP, but they were exceptionally elevated on poor development of ILAC. Most of RMSP became atrophied within 3 months. Delayed-onset bile leak occurred in a donor, by which we had to resect his functional RMSP after 3 months. ILAC was highly associated with persistent bile production and slow atrophic change at RMSP after LLS graft donation. CONCLUSIONS: These data suggested that temporary arterial collateral formation worked as a buffering mechanism leading to slow and uneventful degeneration of inflow-deprived hepatic parenchyma. PMID- 17086887 TI - Phrenic vein dissection in partial liver graft harvesting. AB - BACKGROUND/AIMS: Despite the impressive results of living donor liver transplantation, hepatic venous reconstruction remains a controversial component. METHODOLOGY: A total of 211 consecutive donor hepatectomies were performed. The proximal route of the hepatic vein was exposed by dissection of the connective tissue around the hepatic vein and by dividing and ligating all of the inferior phrenic veins that open into the hepatic vein, into the confluence of the hepatic vein and inferior vena cava, or directly into the inferior vena cava. RESULTS: In the 114 left-side hepatectomy procedures, the number of divided left inferior phrenic veins ranged from 1 to 4 and the diameters of the left and middle hepatic veins ranged from 7 to 33mm. For the 97 right-side procedures, the number of divided right inferior phrenic veins ranged from 1 to 4 and the diameters of right hepatic veins ranged from 9 to 34mm. This maneuver safely allowed for the safe exposure of all trunks and routes of the hepatic veins and the suprahepatic portion of the inferior vena cava. CONCLUSIONS: Our technique is useful for obtaining a wide ostium and a sufficient length of the hepatic vein for grafts obtained from living donors. PMID- 17086888 TI - Model for End Stage Liver Disease score does not predict graft survival after living donor liver transplantation. AB - BACKGROUND/AIMS: Although the Model End-Stage Liver Disease (MELD) score has been effective in the prediction of mortality in patients awaiting liver transplantation, its predictive value of survival after living donor liver transplantation (LDLT) needs further study. METHODOLOGY: We report our experience with 142 cases of LDLT, analyzing the impact of the MELD score on the graft survival after LDLT. RESULTS: Of 142 grafts, 18 failed during the observation period. One-, 3- and 5-year survival rates were 92%, 87% and 82%, respectively. Cox proportional hazard model revealed that MELD score did not predict graft survival (p=0.43, relative risk = 1.0). CONCLUSIONS: The present results may indicate that the preoperative MELD score was of little help for determining the indication of LDLT. PMID- 17086889 TI - Clinicopathologic study of intraductal papillary-mucinous tumors and mucinous cystic tumors of the pancreas. AB - BACKGROUND/AIMS: We analyzed clinicopathologic and imaging findings of intraductal papillary-mucinous tumors (IPMTs) and mucinous cystic tumors (MCTs) of the pancreas to evaluate the difference between IPMTs and MCTs, and to identify the signs indicative of malignancy in IPMTs. METHODOLOGY: Clinicopathological features of 20 patients with IPMT and six patients with MCT of the pancreas were studied. RESULTS: The patients with IPMT comprised 16 males and four females with a mean age of 62.9 years. Eighty percent of IPMTs were located in the pancreatic head, and the mean tumor size was 38.6mm. Recurrence was observed in one patient, who died of IPM adenocarcinoma. In contrast, all patients with MCT were females, with a mean age of 53.0 years. None of the MCTs arose in the pancreatic head, and the mean tumor size was 42.7mm. One patient died of MC adenocarcinoma, but all of the others survived without recurrence. The difference in gender, location of the tumor, and connection to the pancreatic duct reached statistical significance between IPMTs and MCTs. A significant connection to the pancreatic duct and high level of serum carbohydrate antigen 19 9 (CA19-9) was observed in the adenocarcinoma and moderate dysplasia groups of IPMT. CONCLUSIONS: The main duct type and an elevation of serum CA19-9 level suggested malignancy in IPMTs. PMID- 17086890 TI - Significance of pancreatic exocrine function in the perioperative management of pancreatoduodenectomy. AB - BACKGROUND/AIMS: The significance of pancreatic exocrine function in the perioperative management of pancreatoduodenectomy (PD) has not been well understood. The aim of this study was to clarify this issue. METHODOLOGY: Clinical records of 60 Japanese patients who underwent PD were reviewed retrospectively. Patients were divided into two groups, normal (n=33) and low (n=27) pancreatic exocrine function, according to the preoperative value of N benzoyl-L-tyrosyl-p-aminobenzoic acid excretion test (normal value >70%). We compared the perioperative events and nutritional status between the two groups. RESULTS: The preoperative and operative characteristics between the two groups were not significantly different. Postoperative pancreatic juice output from the remnant pancreas during the initial 7 days after PD was greater (1145 +/- 618 vs. 741 +/-612mL, P=0.02), and the prevalence of pancreatic anastomotic leakage was higher (10/23, 30% vs. 1/27, 4%, P=0.008) in the group with normal pancreatic exocrine function than that in the insufficient group. Perioperative body mass index and serum albumin concentration, which reflect the nutritional status of patients, were significantly lower in the group with low pancreatic exocrine function (P=0.007 and 0.04, respectively). CONCLUSIONS: Surgeons should pay more attention to pancreatic anastomotic leakage in patients with normal pancreatic exocrine function after PD. On the other hand, in patients with insufficient exocrine function, perioperative nutritional support should be considered. PMID- 17086891 TI - Comparative study of modified-release clarithromycin and immediate-release clarithromycin in the treatment of Helicobacter pylori-associated peptic ulcer disease. AB - BACKGROUND/AIMS: Proton pump inhibitor-based triple therapy containing immediate release clarithromycin is an important regimen for the eradication of Helicobacter pylori (H. pylori). However, the efficacy of modified-release clarithromycin for the treatment of H. pylori-associated peptic ulcer disease is still unknown. The aims of the study were to compare the efficacy of modified release clarithromycin and immediate-release clarithromycin on the rates of ulcer healing and eradication of H. pylori. METHODOLOGY: One hundred and sixty-one patients with Helicobacter pylori-associated peptic ulcer were randomized to receive one-week triple therapy with either modified-release clarithromycin 1000mg once daily (AECMR) or immediate-release clarithromycin 500mg twice daily (AECIR) in combination with amoxicillin 1,000mg twice daily (A) and esomeprazole 40mg once daily (E). Post-treatment ulcer healing status and Helicobacter pylori status was determined by endoscopy and 13C urea-breath test at 16 weeks and 8 weeks after completion of triple therapy, respectively. RESULTS: Helicobacter pylori eradication rates were 87.5% and 87.7% for AECMR and AECIR, respectively, in the intent-to-treat analysis. Eradication rates in the per-protocol groups were 90.3% and 91.4% for AECMR and AECIR, respectively. In both the intent-to treat and per-protocol analyses, the eradication rates were comparable in the AECMR and AECIR groups (p= 1.0 and 1.0, respectively). Ulcer healing rates in the intention-to-treat analysis were 81.3% and 77.8% for AECMR and AECIR, respectively. Ulcer healing rates in the per-protocol analysis were 90.3% and 90.0% for AECMR and AECIR groups, respectively. In both the intention-to-treat and per-protocol analyses, the ulcer healing rates were comparable in the AECMR and AECIR groups (p=0.645 and 0.584, respectively). CONCLUSIONS: Modified-release clarithromycin 1000mg once daily can be used as an alternative to immediate release clarithromycin 500mg twice daily for the treatment of Helicobacter pylori associated peptic ulcer disease. PMID- 17086892 TI - Influence of COX-2 and local cytokine expressions in gastric ulcer mucosa by H. pylori and NSAID. AB - BACKGROUND/AIMS: H. pylori and NSAIDs are the two most important pathogenic factors in gastric ulcer. NSAIDs and H. pylori share common pathogenic characteristics, but have different effects in modulating COX-2 and local cytokines in gastric ulcer mucosa. This study was designed to explore the influence of COX-2 and local cytokine expressions in gastric ulcer mucosa induced by H. pylori infection and NSAID use. METHODOLOGY: Twenty-three patients were recruited. Sixteen subjects were infected with H. pylori. Fifteen patients used NSAIDs. Gastric biopsy specimens were obtained by endoscopy. COX-2 and local cytokine mRNA expressions were assessed by real-time RT-PCR. RESULTS: COX-2 and local cytokines were over-expressed in gastric ulcer and were positively intercorrelated. H. pylori did not alter COX-2 and local cytokine expressions in gastric ulcer, but induced IL-8 and COX-2 in antral mucosa. NSAIDs inhibited COX 2 expression in gastric ulcer, but had no effect on other local cytokines. COX-2 inhibition by NSAIDs in gastric ulcer was compatible with the findings that NSAIDs delayed gastric ulcer healing. CONCLUSIONS: H. pylori increased IL-8 and COX-2 in the antral mucosa, but did not influence COX-2 and local cytokines in gastric ulcer. NSAIDs inhibited COX-2 in gastric ulcer and delayed gastric ulcer healing. PMID- 17086893 TI - How closely is Helicobacter pylori infection related to gastroduodenal lesions? AB - BACKGROUND/AIMS: Various studies have indicated a relationship between Helicobacter pylori infection and upper gastrointestinal lesions, but this relationship needs to be assessed in individuals not seeking medical treatment for complaints. METHODOLOGY: We screened community residents for H. pylori infection and upper gastrointestinal lesions during an annual mass health examination aiming to determine relationships between infection and lesions. In 932 examinees we performed a 13C-urea breath test for H. pylori infection, and assessed degree of gastric atrophy by measuring pepsinogen I and II in serum. In 738 subjects we also performed upper gastrointestinal radiography with or without endoscopy. RESULTS: Prevalence of H. pylori infection increased with age, and the ratio of serum pepsinogen I to II decreased with age. Prevalence of H. pylori infection did not differ significantly between subjects with and without radiographically or endoscopically evident lesions. Of H. pylori-positive subjects with peptic ulcer, 73.2% had no recurrence of ulcer despite absence of medical treatment. CONCLUSIONS: Prolonged H. pylori infection was associated with atrophy of the gastric mucosa, but little relationship was evident between H. pylori infection and development or recurrence of peptic ulcer. PMID- 17086894 TI - Functional polymorphisms of CD14 and toll-like receptor 4 in Taiwanese Chinese with Helicobacter pylori-related gastric malignancies. AB - BACKGROUND/AIMS: Interindividual differences in degrees and extent of gastritis are responsible for the divergent outcomes after H. pylori infection. Cellular responses in gastric inflammation are mediated by lipopolysaccharide of H. pylori, which activate monocytes to express cytokine expression and growth factors via CD14 and toll-like receptor 4 (TLR4). Whether functional polymorphisms of TLR 4 and CD14 account for H. pylori-related gastric malignancies remains unknown. This study aimed to investigate the contribution of CD14 and TLR4 genotypes to the risk of H. pylori-related gastric malignancies in a Chinese population. METHODOLOGY: Genotyping for CD14 (-159C/T) and TLR4 (Asp 299Gly and Thr 399Ile) was performed in 70 patients with gastric mucosa associated lymphoid tissue lymphoma (MALToma), 204 patients with non-cardia gastric adenocarcinoma (GAC), and 210 unrelated healthy controls. Distribution of genotype and allele frequencies among three groups was compared. RESULTS: The seropositive rate of H. pylori was significantly higher for non-cardia GAC (164/204, 80.4%) and MALToma (66/70, 94.3%) than controls (120/210, 57.5%, p<0.001). A complete absence of Gly or Ile variants was noted for all the studied subjects. The genotype frequencies of CD14 in controls were C/C, 25.7%, C/T, 48.6%, and T/T, 25.7%, and did not deviate from the Hardy-Weinberg equilibrium. The distribution of CD14 genotype did not differ significantly among GAC, MALToma patients, and controls. CONCLUSIONS: These data suggest no apparent association of CD14 polymorphisms with H. pylori-related gastric malignancy and provide evidence for race-specific distribution of TLR4 alleles in Chinese population. PMID- 17086895 TI - Cannabinoids in MS--are we any closer to knowing how best to use them? PMID- 17086896 TI - Retinal ganglion cell loss induced by acute optic neuritis in a relapsing model of multiple sclerosis. AB - Multiple sclerosis (MS) and its animal model experimental autoimmune encephalomyelitis (EAE) are marked by inflammatory demyelinating lesions throughout the central nervous system, including optic nerve. Neuronal loss also occurs in MS and EAE lesions, but it is not known whether neuronal loss occurs secondary to inflammation, or as a primary process. In the current study, the relationship of inflammation to retinal ganglion cell (RGC) loss during acute optic neuritis is examined. RGCs were labelled with Flourogold, and EAE was induced in SJL/J mice by immunization with proteolipid protein peptide 139-151 (PLP). At various time points, RGCs were counted and optic nerves were examined for inflammatory cell infiltrates. No optic neuritis was detected prior to day 9 following immunization. Incidence of optic neuritis was 30% at day 9 and increased to over 70% by day 11, remaining high through day 18. In contrast, no RGC loss was detected in eyes with optic neuritis until day 14. A 43.1% reduction in RGC numbers at day 14 increased to 50.8% by day 18. No RGC loss occurred in eyes without optic neuritis. The fact that inflammation precedes RGC loss suggests that neuronal loss during optic neuritis occurs secondary to the inflammatory process. PMID- 17086897 TI - Elevated urinary excretion of aluminium and iron in multiple sclerosis. AB - Multiple sclerosis (MS) is a chronic, immune-mediated, demyelinating disease of the central nervous system of as yet unknown aetiology. A consensus of opinion has suggested that the disorder is the result of an interplay between environmental factors and susceptibility genes. We have used a battery of analytical techniques to determine if the urinary excretion of i) markers of oxidative damage; ii) iron and iii) the environmental toxin aluminium and its antagonist, silicon, are altered in relapsing-remitting (RRMS) and secondary progressive MS (SPMS). Urinary concentrations of oxidative biomarkers, MDA and TBARS, were not found to be useful indicators of inflammatory disease in MS. However, urinary concentrations of another potential marker for inflammation and oxidative stress, iron, were significantly increased in SPMS (P<0.01) and insignificantly increased in RRMS (P>0.05). Urinary concentrations of aluminium were also significantly increased in RRMS (P<0.001) and SPMS (P <0.05) such that the levels of aluminium excretion in the former were similar to those observed in individuals undergoing metal chelation therapy. The excretion of silicon was lower in MS and significantly so in SPMS (P<0.05). Increased excretion of iron in urine supported a role for iron dysmetabolism in MS. Levels of urinary aluminium excretion similar to those seen in aluminium intoxication suggested that aluminium may be a hitherto unrecognized environmental factor associated with the aetiology of MS. If aluminium is involved in MS then an increased dietary intake of its natural antagonist, silicon, might be a therapeutic option. PMID- 17086898 TI - Interferon-beta modulates bone-associated cytokines and osteoclast precursor activity in multiple sclerosis patients. AB - PURPOSE: Multiple sclerosis (MS) patients have a high risk of low bone density. The purpose of this study was to examine the molecular mechanisms potentially capable of modulating bone homeostasis in response to interferon-beta-1a (IFN beta-1a) treatment and the focus was the bone-modulating system comprised of receptor activator of nuclear factor-kappaB (RANK), its ligand RANKL and its decoy receptor, osteoprotegerin (OPG). METHODS: In this open-label pharmacodynamic study, peripheral blood was obtained from relapsing-remitting MS patients just prior to and at multiple time points after intramuscular injection of 30 microg IFN-beta-1a. Samples were analysed for RANKL, tumour necrosis factor related apoptosis-inducing ligand (TRAIL), OPG and macrophage inflammatory protein-1 alpha/beta expression. Osteoclast precursor differentiation from peripheral blood cells of MS patients in the presence of exogenously added IFN beta-1a was also assessed. Additionally, the changes in plasma levels of osteocalcin and the C-telopeptides after 1 year of treatment were measured as surrogate markers of bone formation and degradation, respectively. RESULTS: IFN beta-1a treatment modulated RANKL and OPG in a selective, time-dependent manner. The levels of OPG protein decreased 25% at the 8-h time point, then increased 43% at the 24-h time point. The levels of free RANKL reached a maximum at the 8-h time point. Increases in the levels of macrophage inflammatory protein-1beta (MIP 1beta), a chemokine that increases osteolysis, were observed. The levels of the bone formation marker, osteocalcin, were lower in MS patients compared to controls and increased after one year of treatment. Ex vivo treatment of peripheral blood lymphocytes with IFN-beta resulted in a marked reduction of osteoclast-like cells in the presence of RANKL and macrophage colony stimulating factor. CONCLUSIONS: IFN-beta treatment induces complex, specific and time dependent changes in multiple proteins and mRNAs related to bone homeostasis in MS patients. PMID- 17086899 TI - Intrathecal chitotriosidase and the outcome of multiple sclerosis. AB - Activated macrophages are major effectors at all stages of lesion formation in multiple sclerosis (MS) brain. Here, we report that the macrophage enzyme chitotriosidase (Chit) is significantly elevated both in plasma and cerebrospinal fluid (CSF) of patients with MS as compared to healthy controls and other neurological patients (P<0.001). Furthermore, the Chit activity in blood significantly associates with the MS clinical course (higher in secondary progressive relative to relapsing-remitting, P=0.01) and the clinical severity as measured by Kurtkze's Expanded Disability Status Scale (P<0.001). Also, we found that Chit activity is compartmentalized in the central nervous system of early MS patients and that its CSF/plasma quotient, in the presence of a preserved albumin quotient, correlates with the extent of future clinical deterioration (r=0.91; P<0.001). These findings confirm that innate immunity, here represented by Chit, is clinically relevant in MS and allows, if confirmed, reconsidering novel MS therapeutic strategies specifically aimed at this branch of the immune response. PMID- 17086900 TI - Production of interferon-gamma by microglia. AB - Neural cells do not usually interact with immune cells because of the lack of major histocompatibility complex (MHC) antigen expression. Interferon-gamma (IFN gamma) enables this interaction via induction of MHC antigen expression in neural cells. Thus, IFN-gamma is a critical cytokine for the development of central nervous system (CNS) pathologies. IFN-gamma, however, is considered to be produced exclusively by lymphoid cells. Here, we show for the first time that murine microglia produce IFN-gamma in response to IL-12 and/or IL-18, using RT PCR detection of IFN-gamma mRNA and Western blotting and immunohistochemical analysis for cytoplasmic expression of IFN-gamma. Stimulation of microglia with IL-12 and IL-18 resulted in MHC class II mRNA expression in microglia. Since IL 12 and IL-18 are produced in the CNS by glial cells, these cytokines may play a critical role in the initiation of neural-immune cell interaction and the induction of autoimmune processes in the CNS via induction of IFN-gamma and MHC antigens. PMID- 17086902 TI - Identifying reliable change in tactile temporal thresholds in multiple sclerosis: test-retest reliability. AB - BACKGROUND: Tactile temporal thresholds are typically significantly higher (ie, prolonged) in multiple sclerosis (MS) patients when compared to controls and increase significantly during relapses, probably reflecting integrity of conduction across a portion of the corpus callosum. As part of an ongoing validation study of tactile temporal thresholds, the test-retest reliability of these thresholds was examined in patients with MS. METHODS: Tactile temporal thresholds were measured in 61 MS patients during two separate test sessions within three weeks. Test-retest reliability and the standard error of measurement were calculated. The threshold of change in tactile temporal thresholds in MS patients that would correspond to real change beyond measurement error with 95% certainty was also calculated. RESULTS: The test-retest reliability of this measure of tactile temporal thresholds was 0.93. The threshold indicating change beyond chance or measurement error with 95% certainty was 19 ms. CONCLUSIONS: This measure of tactile temporal thresholds has excellent test-retest reliability and a change of greater than 19 ms is highly likely to represent real change. This measure is promising as a precise, reliable outcome measure in MS. PMID- 17086901 TI - Classification of multiple sclerosis patients by latent class analysis of magnetic resonance imaging characteristics. AB - BACKGROUND: Disease heterogeneity is a major issue in multiple sclerosis (MS). Classification of MS patients is usually based on clinical characteristics. More recently, a pathological classification has been presented. While clinical subtypes differ by magnetic resonance imaging (MRI) signature on a group level, a classification of individual MS patients based purely on MRI characteristics has not been presented so far. OBJECTIVES: To investigate whether a restricted classification of MS patients can be made based on a combination of quantitative and qualitative MRI characteristics and to test whether the resulting subgroups are associated with clinical and laboratory characteristics. METHODS: MRI examinations of the brain and spinal cord of 50 patients were scored for 21 quantitative and qualitative characteristics. Using latent class analysis, subgroups were identified, for whom disease characteristics and laboratory measures were compared. RESULTS: Latent class analysis revealed two subgroups that mainly differed in the extent of lesion confluency and MRI correlates of neuronal loss in the brain. Demographics and disease characteristics were comparable except for cognitive deficits. No correlations with laboratory measures were found. CONCLUSIONS: Latent class analysis offers a feasible approach for classifying subgroups of MS patients based on the presence of MRI characteristics. The reproducibility, longitudinal evolution and further clinical or prognostic relevance of the observed classification will have to be explored in a larger and independent sample of patients. PMID- 17086903 TI - The Italian Multiple Sclerosis Database Network (MSDN): the risk of worsening according to IFNbeta exposure in multiple sclerosis. AB - We evaluated the risk of worsening according to the length of exposure to interferon beta (IFNbeta) in a large cohort of 2090 multiple sclerosis patients collected by the Italian MS Database Network. Overall 44,140 patient-visits with a follow-up of 22,143 patient-years were evaluated. Forty-one per cent of patients were exposed to IFNbeta for up to 2 years, 39% for 2-4 years and 20% for more than 4 years. A Cox regression model was used to analyse two clinical outcomes: disability progression and worsening of relapse rate. The technique of propensity score was applied to reduce bias in the comparison of non-randomized groups. The risks of disability progression (HR =0.23; 95% CI: 0.17-0.30) and worsening of relapse rate (HR =0.19; 95% CI: 0.14-0.27) were reduced by about 4-5 fold in patients exposed to IFNbeta for more than four years, compared with patients exposed for up to two years. The propensity score technique confirmed the findings. The proportion of days covered by IFNbeta treatment was lower (P <0.0001) in patients exposed to IFNbeta for up to two years than in other groups. A clinical stabilization over two years of IFNbeta exposure may predict a subsequent good clinical response to treatment. PMID- 17086904 TI - The PASAT performance among patients with multiple sclerosis: analyses of responding patterns using different scoring methods. AB - The Paced Auditory Serial Addition Test (PASAT) is widely used in the evaluation of multiple sclerosis (MS) patients' cognitive performance, and also used as the sole measure of cognition in a recently developed assessment tool for MS clinical trials, the Multiple Sclerosis Functional Composite (MSFC). We analysed if MS patients and healthy controls have different patterns of responding in the PASAT, and whether different scoring methods influence the PASAT's sensitivity and specificity in detecting disease-associated cognitive impairment. Forty-five relapsing-remitting MS patients and 48 healthy controls were evaluated using the PASAT and a comprehensive neuropsychological examination. Cognitively deteriorated MS patients compensated for their difficulties in PASAT by omitting rather than guessing answers. They skipped items intermittently, which reduces the difficulty of the task. Furthermore, towards the end of the PASAT's 60-item series MS patients' performance had a trend to fade whereas controls' performance was more even throughout the task. The dyad score or the percent dyad score did not essentially improve the sensitivity or the specificity, but the accuracy improved when the answers at the end of the PASAT series were specifically emphasized. Using the combined score, 73% of the patients were correctly classified as cognitively impaired or unimpaired. PMID- 17086905 TI - Clinical impact of 20% worsening on Timed 25-foot Walk and 9-hole Peg Test in multiple sclerosis. AB - INTRODUCTION: Quantitative tests of motor function, like the Timed 25-foot Walk (T25FW) and 9-hole Peg Test (9HPT), are increasingly being applied as outcome measures in multiple sclerosis (MS) clinical trials. The quantitative nature of the data has a favorable impact on responsiveness, but the clinical impact of the changes is uncertain. The goal of this study was to assess whether a change on T25FW and 9HPT does indeed have a clinical meaning. This was accomplished by comparing 20% changes on these quantitative measurements to concomitant changes on the Guy's Neurological Disability Scale (GNDS), a scale which measures patient perceived daily life disability. METHODS: From a longitudinal database, we selected patients with at least two measurements of T25FW, 9HPT and GNDS with a minimal time interval of 350 days. In those patients who experienced at least a 20% change on T25FW or 9HPT, GNDS score changes were examined more closely. RESULTS: Of 527 patients, 143 experienced a >20% worsening on their T25FW and 71 on their 9HPT, respectively. Patients with a 20% increase in T25FW or 9HPT had more GNDS worsening than patients without such an increase. GNDS worsening associated with an increase in T25FW was mainly due to an increase in perceived disability related to lower extremity function and fatigue; GNDS worsening associated with an increase in 9HPT was more diffuse with respect to domains involved. CONCLUSION: Worsening on T25FW or 9HPT has a clinical impact on disability, as perceived by MS patients during daily life functioning. PMID- 17086906 TI - The Face-Symbol Test and the Symbol-Digit Test are not reliable surrogates for the Paced Auditory Serial Addition Test in multiple sclerosis. AB - The Paced Auditory Serial Addition Test (PASAT) is the chosen task for cognitive assessment in the multiple sclerosis functional composite (MSFC) and a widely used task in neuropsychological studies of people with multiple sclerosis (MS), but is unpopular with patients. The Face-Symbol Test (FST) and Symbol-Digit Tests (SDT) are alternative methods of cognitive testing in MS, which are easily administered and patient-friendly. In order to evaluate the potential of the FST as a possible surrogate for the PASAT, we directly compared the FST to the PASAT and the SDT in a cohort of 50 MS patients with varying levels of disability. There was significant correlation between SDT and FST scores (Spearman's rho 0.80, 95% CI 0.66-0.88), R(2) 65%, with moderate inter-test agreement (k =0.52). In contrast, SDT and FST scores were less predictive of PASAT scores. We concluded that neither the FST nor SDT are reliable surrogates for the PASAT. PMID- 17086907 TI - Health-related quality of life in a population-based sample of people with multiple sclerosis in Stockholm County. AB - The aim of this descriptive, cross-sectional study was to analyse health-related quality of life (HRQoL) in a population-based sample of people with multiple sclerosis (PwMS) in Stockholm County, with respect to disease-related and sociodemographic factors and coping capacity. A further aim was to compare our results on HRQoL--collected by face-to-face interviews at home-visits--with the results from a study with a mail-surveyed sample of PwMS in Stockholm. Home visits were made to 166 PwMS with clinically definite MS. Data were obtained from structured interviews using the Sickness Impact Profile (SIP), EuroQol-5D (EQ-5D) and the Sense of Coherence Scale. HRQoL was widely affected, especially in home management, walking and recreation. Self-rated HRQoL was worse in PwMS, including those with milder disease and shorter disease duration, than in the general population. Factors that were independently associated with large impact on HRQoL were disease severity, work status and coping capacity. The EQ-5D score of PwMS was more favourable when assessed by face-to face interviews at home in our study than in the study using mailed questionnaires. The study gives detailed information on HRQoL and will contribute to the base needed for organizing health care services aimed at improving HRQoL of PwMS in Stockholm. PMID- 17086908 TI - Gait variability in people with multiple sclerosis. AB - Patients with multiple sclerosis (MS) present with varying symptoms that can differ between and within individuals. As new interventions and drug treatments become available to MS patients, it is essential to understand the relationship between fatigue and the variability of functional mobility measures in order to define a meaningful change due to treatment within the MS population. The purpose of this study was to examine the within-day and between-day changes in gait variability for subjects with MS in fresh and fatigued conditions. Walking gait parameters were measured from 20 subjects diagnosed with MS and eight healthy control subjects. Standard deviations of hip, knee and ankle kinematic and kinetic variables were quantified as the measure of variability and analysed with a two-way (group by condition) ANOVA. Results indicated MS subjects had significantly greater hip (P <0.020), knee (P <0.011) and ankle (P <0.034) joint angle variability than control subjects, but variability was not different between conditions. Kinetic variability was not different between groups or conditions. MS subjects in this study walked more slowly than the healthy controls and they also reported more fatigue. Research examining treatment effects within the MS population should account for increased levels of kinematic gait variability. PMID- 17086909 TI - Gait and balance impairment in early multiple sclerosis in the absence of clinical disability. AB - This study evaluated the gait and balance performance of two clinically distinct groups of recently diagnosed and minimally impaired multiple sclerosis (MS) patients (Expanded Disability Status Scale range 0-2.5), compared to control subjects. Ten MS patients with mild pyramidal signs (Pyramidal Functional Systems 1.0), 10 MS patients with no pyramidal signs (Pyramidal Functional Systems 0) and 20 age- and gender-matched control subjects were assessed using laboratory-based gait analysis and clinical balance measures. Both MS groups demonstrated reduced speed and stride length (P < 0.001), and prolonged double limb support (P <0.02), compared to the control group, along with alterations in the timing of ankle muscle activity, and the pattern of ankle motion during walking, which occurred independent of gait speed. The pyramidal MS group walked with reduced speed (P = 0.03) and stride length (P = 0.04), and prolonged double limb support (P =0.01), compared to the non-pyramidal group. Both MS groups demonstrated concomitant balance impairment, performing poorly on the Functional Reach Test compared to the control group (P <0.05). The identification of incipient gait and balance impairment in MS patients with recent disease onset suggests that motor function may begin to deteriorate in the early stages of the disease, even in the absence of clinical signs of pyramidal dysfunction. PMID- 17086910 TI - The scope and nature of pain in persons with multiple sclerosis. AB - Much remains unknown about the scope, nature, and impact of pain on the lives of persons with multiple sclerosis (MS). In the present study, 180 community dwelling adults with MS completed a postal survey that included demographic measures, MS disease measures, and several standardized measures of pain, including pain intensity, variability, location, and pain-related interference. Some 66% of the sample reported pain, 25% of whom reported severe pain. Persons with pain reported an average of 6.6 distinct pain sites. Using the Brief Pain Inventory Interference Scale, the average level of overall pain interference was 3.33 (0-10 scale) in the group reporting pain. The highest levels of pain interference were reported for sleep, recreational activities, and work in and outside the home. Persons with pain were more likely to report greater MS disease severity, poorer psychological functioning, and poorer health than persons with MS but not pain. Persons with pain were also less likely to be employed. These findings are consistent with previous research that shows that pain is common in MS, that it is severe in a substantial subset of these individuals and has the potential to negatively impact physical and psychosocial functioning over and above the effects of MS itself. PMID- 17086911 TI - Long-term use of a cannabis-based medicine in the treatment of spasticity and other symptoms in multiple sclerosis. AB - The object of this study was to monitor the safety and efficacy of long-term use of an oromucosal cannabis-based medicine (CBM) in patients with multiple sclerosis (MS). A total of 137 MS patients with symptoms not controlled satisfactorily using standard drugs entered this open-label trial following a 10 week, placebo-controlled study. Patients were assessed every eight weeks using visual analogue scales and diary scores of main symptoms, and were followed for an average of 434 days (range: 21 -814). A total of 58 patients (42.3%) withdrew due to lack of efficacy (24); adverse events (17); withdrew consent (6); lost to follow-up (3); and other (8). Patients reported 292 unwanted effects, of which 251 (86%) were mild to moderate, including oral pain (28), dizziness (20), diarrhoea (17), nausea (15) and oromucosal disorder (12). Three patients had five 'serious adverse events' between them--two seizures, one fall, one aspiration pneumonia, one gastroenteritis. Four patients had first-ever seizures. The improvements recorded and dosage taken in the acute study remained stable. Planned, sudden interruption of CBM for two weeks in 25 patients (of 62 approached) did not cause a consistent withdrawal syndrome, although 11 (46%) patients reported at least one of--tiredness, interrupted sleep, hot and cold flushes, mood alteration, reduced appetite, emotional lability, intoxication or vivid dreams. Twenty-two (88%) patients re-started CBM treatment. We conclude that long-term use of an oromucosal CBM (Sativex) maintains its effect in those patients who perceive initial benefit. The precise nature and rate of risks with long-term use, especially epilepsy, will require larger and longer-term studies. PMID- 17086912 TI - Cannabis use in patients with multiple sclerosis. AB - INTRODUCTION: Little is known about the extent and patterns of cannabis use in people with multiple sclerosis (MS). METHODS: MS patients attending neurology outpatient clinics at two hospitals in London and one in Kent, UK completed a questionnaire. RESULTS: Questionnaires were completed by 254/337 (75%) MS patients. Forty-three per cent had used cannabis at some stage (ever users). Of these, 68% (75/110) had used cannabis to alleviate symptoms of MS (MS-related cannabis use). Forty-six (18%) had used cannabis in the last month (current users), of whom 12% (31/254) had used it for symptom relief. Being married or having a long-term partner, tobacco smokers and increasing disability were independent risk factors for MS-related cannabis use. Compared to patients who could walk unaided, cannabis use was more likely in those who were chair-bound (adjusted OR 2.47; 1.10-5.56) or only able to walk with an aid (adjusted OR 1.56; 0.90-3.60). Pain and spasms were common reasons for cannabis use. Seventy-one per cent of individuals who had never used cannabis said they would try the drug if it were available on prescription. CONCLUSION: A large proportion of MS patients had tried cannabis for symptom control, however current use was small. A subgroup with greater disability appears to derive some symptomatic benefit. PMID- 17086913 TI - Serum immunologic markers in multiple sclerosis patients on continuous combined therapy with beta-interferon 1a, prednisone and azathioprine. AB - Break-through symptoms (BTS) in multiple sclerosis (MS) patients on beta interferon (beta-IFN) monotherapy are most frequently treated with a brief administration of steroids. Here, we report the results of monitoring serum immunologic markers recorded at three-month intervals for 1.5 years in responders to beta-INF 1a (Avonex) monotherapy (n =21) and MS patients placed on Avonex with prednisone (n =83) and Avonex, prednisone and azathioprine (AZA) (n =21) because of BTS. Compared to 23 healthy controls, patients on Avonex monotherapy and Avonex with prednisone, in individuals on Avonex, prednisone and AZA, a significant decrease in serum concentration of soluble intercellular adhesion molecule-1 (sICAM-1) (P=0.001) was established. Combined therapy with Avonex, prednisone and AZA was associated with a significant increase in the serum level of interleukin (IL)10 (P <0.001). Compared to Avonex monotherapy, combined therapy suppressed the serum level of IL12p40, antagonized elevation in the serum concentration of soluble IL2 receptor (sIL2R) and inhibited an increase in the serum soluble CD95 (sCD95) molecule. In patients studied, no significant differences in the serum level of IL18 and tumor necrosis factor-alpha (TNF alpha) were established. These findings are important in understanding some of the immunoregulatory mechanisms induced by combined therapy in MS. PMID- 17086915 TI - Cervical cord magnetization transfer ratio and clinical changes over 18 months in patients with relapsing-remitting multiple sclerosis: a preliminary study. AB - BACKGROUND: Magnetization transfer ratio (MTR) permits the quantitative estimation of cervical cord tissue damage in patients with multiple sclerosis (MS). OBJECTIVE: To determine whether a single time-point MTR scan of the cervical cord is associated with short-term disease evolution in patients with relapsing-remitting (RR) MS. METHODS: Using a 1.5-T magnetic resonance imaging (MRI) system with a tailored cervical cord phased array coil, fast short-tau inversion recovery (fast-STIR) and MTR scans were obtained from 14 untreated patients with RRMS at baseline. Cervical cord MTR histograms were derived. Over the 18-month follow-up period, relapse rate was measured and disability assessed by the Expanded Disability Status Scale (EDSS) score. RESULTS: Average cervical cord MTR was correlated with relapse rate (r= -0.56, P=0.037). A moderate correlation (r values ranging from -0.33 to -0.36) between baseline cervical cord MTR metrics and EDSS changes over 18 months was also noted, albeit statistical significance was not reached (P = 0.26 and 0.21, respectively) perhaps because of the relatively small sample size. CONCLUSIONS: This study suggests that a 'snapshot' MT MRI assessment of the cervical cord may detect cervical cord tissue changes associated with short-term disease evolution in RRMS. PMID- 17086914 TI - Prevalence of oedema of the lower limbs in multiple sclerosis patients: a vascular and lymphoscintigraphic study. AB - The aim of the study was to evaluate the frequency of oedema of the lower limbs in multiple sclerosis (MS) patients utilizing a multidisciplinary approach. A total of 205 patients with definite MS were included in the study. Seventy-five were male and 130 female, with a mean age of 50.53, mean Expanded Disability Status Scale (EDSS) score of 5.27 and mean disease duration of 16.6 years. Seventy-one patients had a relapsing-remitting (RR) disease course, 85 were secondary progressive (SP) and 49 were primary progressive (PP). Ninety-three patients (45%) showed oedema at the examination. EDSS, disease duration and disease course, but not gender, were statistically different between oedema and non-oedema patients. Out of 93 patients with oedema, 69 agreed to undergo a vascular examination. Of 69 patients, 45 (65.2%) had a CEAP score (specific rating scale for oedema) of 3 (presence of oedema) and 24 (34.8%) had a score of 4 (presence of a trophic disorder). Out of 69 subjects, 33 agreed to undergo a lymphoscintigraphy, which was normal in only 29 extremities out of 66. Lower limb oedema is common in MS patients, especially in those with reduced mobility. Early screening is advised in patients with an elevated EDSS. PMID- 17086916 TI - Improvements in muscle-tendon properties are beneficial to balance in multiple sclerosis. AB - A 63-year-old male was diagnosed with multiple sclerosis (MS) two years prior to this investigation. We hypothesized that compromised postural balance in MS can be improved by training-induced alterations in muscle-tendon properties. Sixteen weeks of strength and balance training induced an increase in quadriceps and gastrocnemius muscle size, strength, central activation, muscle recruitment and tendon stiffness. Concomitantly, the ability to maintain postural stances was increased in both eyes-open and eyes-closed conditions. We concluded that improvements in muscle-tendon properties were accompanied by improved balance and a general feeling of well-being after training. PMID- 17086917 TI - Recurrent optic neuromyelitis with endocrinopathies: a new syndrome or just a coincidence? AB - The spectrum of optic neuromyelitis (ONM) ranges from monophasic or recurrent idiopathic forms of the disease, to ONM associated with autoimmune disorders. A distinct form of the disease, called recurrent ONM with endocrinopathies, characterized by spinal cord involvement (cavitations with syringomyeloid sensory disturbance), rapid evolution to blindness and paraplegia, characteristic cerebrospinal fluid (CSF) findings, and association with hypothalamus-pituitary dysfunction, has recently been described. The first case of ONM with endocrinopathies in a female Caucasian from Europe is presented, supporting the existence of this syndrome as a separate entity. PMID- 17086918 TI - Failure to detect JC virus DNA in cerebrospinal fluid of multiple sclerosis patients. PMID- 17086919 TI - Putting limits on teen drivers. PMID- 17086920 TI - Wanless warning: future of NHS hangs on obesity action. PMID- 17086921 TI - NICE insiders criticise Alzheimer's drug decision. PMID- 17086922 TI - NHS employers: far fewer job cuts than first claimed. PMID- 17086924 TI - Five years and billions of pounds later, has anything changed? PMID- 17086923 TI - Child visitors face ban. PMID- 17086925 TI - On long tails and J curves. PMID- 17086926 TI - Data briefing. Length of stay--the day-case drive. PMID- 17086927 TI - Equal, but not the same. AB - In April 2007, the gender equality duty will make it obligatory for all health providers to actively demonstrate equity in service provision. Good practice tends to exist in small projects with little evidence of national progress towards gender equity. The DoH says trusts should already be working towards PMID- 17086929 TI - People power. PMID- 17086928 TI - On financial influence. PMID- 17086930 TI - The neuropeptide Y5 receptor as a therapy for obesity--is it time for a re assessment? PMID- 17086931 TI - Selective androgen receptor modulators: in pursuit of tissue-selective androgens. AB - The androgen receptor mediates the androgenic and anabolic activity of the endogenous steroids testosterone and 5alpha-dihydrotestosterone. Current knowledge of the androgen receptor protein structure, and the molecular mechanisms surrounding the binding properties and activities of agonists and antagonists has led to the design and development of novel nonsteroidal ligands with selected tissue-specific androgen receptor agonist and antagonist activities. The activity of these compounds, termed selective androgen receptor modulators (SARMs), is directed toward the maintenance or enhancement of anabolic effects on bone and muscle with minimal androgenic effects on prostate growth. SARMs are of potential therapeutic value in the treatment of male hypogonadism, osteoporosis, frailty and muscle wasting, burn injury and would healing, anemia, mood and depression, benign prostatic hyperplasia and prostate cancer. PMID- 17086932 TI - Progesterone receptor antagonists. AB - Since the discovery of the progesterone receptor antagonist mifepristone, numerous additional compounds, which display a spectrum of biological actions ranging from full antagonist to those with mixed agonist/antagonist activity, have been synthesized. The latter are referred to as selective progesterone receptor modulators. Long-term administration of these agents is associated with an antiproliferative action on the endometrium as well as amenorrhea and often inhibition of ovulation. Thus far, the majority of clinical data have been obtained with mifepristone but studies are currently underway with other compounds. These compounds have application in the treatment of uterine myoma, endometriosis, dysfunctional uterine bleeding, as potential contraceptives and in steroid responsive tumors. PMID- 17086934 TI - Modulation of cyclic nucleotides and cyclic nucleotide phosphodiesterases in pancreatic islet beta-cells and intestinal L-cells as targets for treating diabetes mellitus. AB - Cyclic 3'5'-AMP (cAMP) is an important physiological amplifier of glucose-induced insulin secretion by the pancreatic islet beta-cell. In the beta-cell, cAMP is formed by the activity of adenylyl cyclase, especially in response to the incretin hormones glucagon-like peptide (GLP)-1 and glucose-dependent insulinotropic peptide. cAMP may also play a similar role in regulating GLP-1 secretion from intestinal L-cells. cAMP influences many steps involved in glucose induced insulin secretion and may be important in regulating pancreatic islet beta-cell differentiation, growth and survival. cAMP itself is rapidly degraded in the pancreatic islet beta-cell by cyclic nucleotide phosphodiesterase enzymes. This review will discuss the possibility of targeting cAMP mechanisms in the treatment of type 2 diabetes mellitus, in which insulin release in response to glucose is impaired. PMID- 17086933 TI - Insulin resistance and PPAR insulin sensitizers. AB - Drugs that reverse insulin resistance are of importance as insulin resistance is frequently associated with type 2 diabetes. The three peroxisome proliferator activated receptors (PPARs) PPARalpha, PPAR90 and PPARgamma are essential for the actions of the many insulin sensitizers. PPARalpha activation enhances free fatty acid oxidation and potentiates anti-inflammatory effects, while PPARgamma is essential for normal adipocyte differentiation and proliferation, as well as fatty acid uptake and storage. Thiazolidinediones (TZDs) are selective ligands of PPARgamma and act as insulin sensitizers. TZDs also suppress free fatty acids via the inhibition of lipolysis in adipose tissue. Insulin sensitizers currently under development include partial PPARgamma agonists and antagonists, and dual PPARalpha/PPARgamma agonists. Given that TZDs show anti-inflammatory, anti oxidant and antiprocoagulant properties in addition to their insulin sensitizing and antilipotoxic properties, a case may be made for initiating TZD therapy early in the treatment of type 2 diabetes, particularly in those patients at risk of cardiovascular disease. TZDs may also be an important therapeutic option in the treatment of metabolic syndrome. PMID- 17086935 TI - Vitamin D receptor ligands for osteoporosis. AB - 1alpha,25-Dihydroxyvitamin D3 (1,25-(OH)2D3), the biologically active metabolite of vitamin D, mediates its actions via the vitamin D receptor (VDR), a member of the superfamily of steroid/thyroid hormone/retinoid receptors. 1,25-(OH)2D3 is required for calcium and phosphorus homeostasis, and for normal skeletal development as well as maintenance of skeletal architecture. Two VDR ligands, calcitriol (1,25-(OH)2D3) and its synthetic analog alfacalcidol (1alpha hydroxyvitamin D3), have been approved for the treatment of osteoporosis. However, the use of calcitriol and alfacalcidol is limited by their major side effect, hypercalcemia, which is mediated mainly by VDR activity in the small intestine. In order to identify VDR ligands with less hypercalcemia liability, a number of pharmaceutical companies are pursuing efforts to develop synthetic vitamin D analogs. This review discusses the mechanism of action of vitamin D, and summarizes the currently approved anti-osteoporotic VDR agonists and compounds that are under development. The future directions of vitamin D research for the discovery of novel VDR agonists for osteoporosis are also discussed. PMID- 17086936 TI - Vitamins: not just for enzymes. AB - Vitamins have traditionally played the role of coenzymes, organic molecules that facilitate the chemical reactions catalyzed by enzymes. However, several vitamins assume additional endocrine-like actions; this review will discuss four such vitamins. Vitamin K2 is involved in the gamma-carboxylation of coagulation factors and bone proteins, but it can also bind and activate the steroid and xenobiotic receptor in order to mediate transcription in bone tissue, and has been used to treat osteoporosis. Biotin is critical for some carboxylation reactions, but it also induces epidermal differentiation and has been used to treat lameness in animals and brittle nails in humans. Pyridoxal phosphate (the active form of vitamin B6) is involved in a multitude of reactions, including decarboxylation and transamination; it can also inhibit DNA polymerases and several steroid receptors and may prove useful as an adjunct in cancer chemotherapy. Finally, nicotinic acid is converted to NAD+ and NADP+, which are used as hydrogen/electron carriers in redox reactions. However, it also possesses vasodilatory and antilipolytic activities. PMID- 17086937 TI - Drug evaluation: ranirestat--an aldose reductase inhibitor for the potential treatment of diabetic complications. AB - Ranirestat is an orally available aldose reductase inhibitor under development by Dainippon Sumitomo Pharma Co Ltd (formerly Dainippon Pharmaceutical Co Ltd), Kyorin Pharmaceutical Co Ltd and Eisai Co Ltd for the potential treatment of diabetic complications, such as neuropathy, cataracts, retinopathy and nephropathy. Phase III trials were ongoing in February 2006. PMID- 17086938 TI - Drug evaluation: tagatose in the treatment of type 2 diabetes and obesity. AB - Spherix Inc (formerly Biospherics) is developing tagatose, an orally active lactose derivative for the potential treatment of obesity and type 2 diabetes. The compound is also under investigation for the potential treatment of anemia, hemophilia and medical problems related to infertility, birth weight and excessive maternal food intake. Phase I and II clinical trials have been completed. PMID- 17086939 TI - Drug evaluation: tesamorelin, a synthetic human growth hormone releasing factor. AB - Theratechnologies, under license from Valeant, is developing tesamorelin as a potential vaccine adjuvant and for the potential treatment of wasting, hip fracture recovery, immune disorders, HIV-related lipodystrophy, sleep maintenance insomnia and mild cognitive impairment. Phase III clinical trials for the treatment of HIV-associated lipodystrophy and phase II clinical trials for sleep disorder, chronic obstructive pulmonary disorder, hip fracture and immune system dysfunction are underway. Phase II trials are also assessing the influenza vaccination immune response and cognitive effects of tesamorelin. PMID- 17086942 TI - [Short-term psychodynamic psychotherapy]. AB - BACKGROUND: The past twenty years have seen many developments in 'psychodynamic' psychotherapy. AIM: To provide an overview of recent developments in short-term psychodynamic psychotherapy. This involves summarizing current approaches and describing in detail the model developed by Malan and Davanloo. A summary is also given of empirical research into the effect of the various treatment modalities. METHOD: Modern manuals were consulted, which describe many of the recent developments and contain contributions by the leading representatives of the various therapeutic orientations. In addition, a literature survey was carried out; by means of Webspirs it was possible to search PsycINFO, Medline and Current Contents for studies relating to empirical research in the effect of psychodynamic psychotherapy. RESULTS There are several variants of the psychodynamic treatment modality. In each of them the treatment duration is reduced, mainly because the focus is on certain particular elements. In addition, a maximum number of sessions is specified. The frequency can be varied and the psychotherapist is much more actively involved than in classical psychoanalytic psychotherapy. Techniques from other therapeutic orientations are integrated, but the focus on defence and affect expressions is preserved. Especially shorter treatment has stimulated empirical research and gradually it is becoming possible to compare short-term treatment with other forms of treatment. CONCLUSION: Short term psychodynamic psychotherapy fits in with modern requirements with regard to efficiency, brevity and (empirical) transparency. This form of treatment deserves its own place in modern psychotherapy because it focuses directly on the patient's actual feelings and emotions. Research into the effectiveness of dynamic therapy has already begun, but more work needs to be done in this area. PMID- 17086941 TI - [Measurement of quality. Comment on Serdobbel et al]. PMID- 17086940 TI - [Obsessive compulsive disorder and the internet. An evaluation of Dutch-language websites and quality indicators]. AB - BACKGROUND: The Internet is an important source of information about health matters. The quality of the information about Obsessive Compulsive Disorder (OCD) has not been investigated. Possible indicators of quality are the DISCERN rating tool, Google PageRank and cooperation from health care professionals. AIM: To investigate the quality of the information given on Dutch-language websites relating to OCD and to determine whether the quality of the contents is correlated with site characteristics, scores on DISCERN and Google PageRank. METHOD: We selected 27 Dutch-language websites via Google and Ixquick and investigated the correlation between the quality of the contents measured by 4 parameters and the presence of incorrect information, site characteristics, the DISCERN score and Google PageRank. RESULTS: The quality of the information was generally poor. The only investigated site characteristic that was found to be associated with basic criteria relating to symptoms and diagnosis was the size of the site. The DISCERN score was significantly correlated with the quality of the information. The occurrence of incorrect information was independent of the quality of the content, site characteristics, Google PageRank and DISCERN scores. CONCLUSION: The quality of Dutch-language websites on the subject of OCD is inadequate. Site characteristics and Google PageRank are not good predictors of the quality of information. The DISCERN rating tool is a potentially valuable indicator of content quality with regard to treatment options. PMID- 17086943 TI - [Sharing information among carers involved in mental health care: ethical advice]. AB - This article reproduces ethical advice given by the Ethical Committee for Mental Health Care set up by the Brothers of Charity in Flanders. The advice relates to the ethical problem that arises when several health care professionals and carers cooperate in mental health care: when and how should they share information about a patient or should information be kept confidential? The research on which the advice was based, consists of discussing ethical matters and studying the relevant literature. Two general principles emerged from the research, namely, consultation and carefulness. These principles are applicable in all situations. On every occasion the involved professionals and carers test whether patient information should be passed on. The test is conducted on the basis often carefulness criteria. The next step is to apply these general principles. If a team of carers is involved, then, it is argued, there should be shared confidentiality, the relevance of which undergoes a filtering process. On the other hand, when there is a network of professionals and carers and third parties are involved, consultation and carefulness should prevail, although exceptions can be made in certain circumstances. PMID- 17086944 TI - [Information about electroconvulsive therapy on the internet]. AB - This article aims to provide a quantitative and qualitative assessment of the information about electroconvulsive therapy that is currently available on the internet. We carried out a quantitative assessment by entering five search terms into eight (meta)search engines. We achieved our qualitative assessment by visiting the first twenty websites generated by each search on one of the search engines, in particular Google (www.google.com), and by scoring these websites with an adapted Sandvik-score. We conclude that the scored websites are technically sound but are incomplete as far as content is concerned. PMID- 17086945 TI - [Cyber-suicide: the role of the internet in suicidal behaviour. A case study]. AB - The article describes two cases of attempted suicide which were prepared and carried out with the help of information on the internet. It appears therefore that the internet can lower the threshold for and increase the risk of suicidal behaviour. The internet can provide the necessary information and at the same time trigger imitation and interaction. In addition, however, the internet can play a positive role by helping to prevent suicide. PMID- 17086946 TI - [Psychoses and epilepsy. A case study]. AB - Over the last few decades much research has been done into the raised level of psychiatric comorbidity in epilepsy. On the basis of a case study of a patient suffering from post-ictal psychoses we explain the psychiatric differential diagnosis within the framework of epilepsy and we investigate the frequent psychiatric side-effects of anticonvulsants. It is concluded that the links between epilepsy and psychiatric symptoms are complex and that the neuropsychiatry of epilepsy is concerned with syndromes that are unique and do notfit into modern psychiatric classification systems. PMID- 17086947 TI - [Reaction to 'The psychiatrist working in a multidisciplinary team and the "Revised Profile of the Psychiatrist." Legal and organizational statements outside reality?']. PMID- 17086948 TI - Texas State Board of Dental Examiners approves rule changes and new "Jurisprudence Assessment". PMID- 17086949 TI - David vs. Goliath. PMID- 17086950 TI - Oral and maxillofacial pathology case of the month. Squamous papilloma. PMID- 17086951 TI - Guilty in Rhode Island. Conviction of hospital executives in Roger Williams case strengthens the argument that stronger oversight needed. AB - The conviction of two former Roger Williams Medical Center executives in Rhode Island is sure to keep up the heat for more vigorous oversight of hospitals. Robert Urciuoli and Frances Driscoll were both convicted of fraud. "It's not to say that directors should be running these organizations," says attorney Patrick Coffey, left. "But it is their function to hire qualified people (who) won't ever tarnish the reputation of their institution". PMID- 17086952 TI - EHR adoption 'pitifully behind'. Study: only 10% of physicians use IT to its fullest. PMID- 17086953 TI - Providers fight CMS' quality initiative. Controversial proposal would cut outpatient pay for not reporting inpatient data. PMID- 17086954 TI - Safety in numbers. Leapfrog lists top hospitals; some question usefulness. PMID- 17086955 TI - In for the long haul. PHRs, computer-assisted coding touted at AHIMA. PMID- 17086956 TI - Objective observers? Fraud cases debunk belief that whistle-blowers are insiders. PMID- 17086957 TI - The next big leap. Hospital ranking, P4P effort up the ante in bid to spur quality of care. PMID- 17086958 TI - Police thyself. From GPOs and QIOs to not-for-profit hospitals, leaders hope a voluntary-compliance approach can stave off more regulation. PMID- 17086959 TI - Controversial move. Leavitt seeks to tie genetic info to e-health records. PMID- 17086960 TI - How to read a face. PMID- 17086961 TI - [New immunology--immunology of pattern recognition receptors]. AB - Pattern recognition receptors (PRRs) have been found on all cells of the body- cells of the innate and adaptive immune systems, epithelial and endothelial cells, keratinocytes, etc. PRRs can recognize specific molecular structures of microorganisms as well as allergens and other substances. The interaction with ligands of foreign microorganisms activates PRRs, after which host cells start to produce cytokines to both specifically activate innate immunity and to control adaptive immune reactions. On the other hand, no immune response develops against microorganisms of the normal microflora. Practically, the development of all immune responses is controlled by PRRs. These responses start in epithelial cells, skin cells, and vascular epithelial cells, which meet alien first. The immune system uses these cells to control the composition of normal microflora. Accordingly, the definition of immune system functions should be complemented by the regulation of body's microflora in addition to the protection from alien and altered self. PMID- 17086962 TI - [The problem of organism integrity and its perspectives]. AB - The organism integrity in onto- and phylogenesis is considered as an initial element of biosphere integrity and its evolution at the eukaryotic level. The Schmalhausen's concept of organism integrity is discussed in the context of his original strategy of evolutionary synthesis and his works on stabilizing selection. The perspectives of investigations on the problem of integrity and development of the principle of dynamic stability in contemporary evolutionary biology as integrating factors are discussed. PMID- 17086963 TI - [Biological functions of allantoin]. AB - A historical review of allantoin research is presented. An increased allantoin level in the trophoblast and serum of pregnant women has been demonstrated. Allantoin concentration decreased in placental tissues and increased in the serum in developing placental insufficiency. PMID- 17086964 TI - [The mechanisms of nitric oxide production from exogenous and endogenous NO donating compounds and its effect on deoxyribonucleotide and DNA synthesis]. AB - The mechanisms of exogenous nitric oxide (NO) production through in vivo biotransformation of nitro-, nitroso- and amino-containing substances were discussed. In addition, the mechanisms of production and cellular sources of endogenous NO, appearing in the blood and tissues after the exposure to various DNA-damaging factors, have been considered. Considerable quantities of endogenous NO were detected in the body in the first hours after translation inhibition by cycloheximide or animal exposure to superlethal radiation doses, i.e., after the exposure to factors inducing destructive processes. The time and dose dependences of exogenous and endogenous NO production have been established. NO produced after a single or repeated administration of NO-donating compounds as well as endogenous NO proved to inhibit deoxyribonucleotide (dNTP) and DNA synthesis in animal tissues. Nonspecific compensatory responses to disturbed protein homeostasis included cyclic production of endogenous NO. The maximum levels of nitrosyl complexes were registered when the rate of protein synthesis decreased. The role of polyamines in the induction of macromolecule biosynthesis is discussed and NO production from these arginine-rich compounds is proposed. NO is released at the stage of polyamine inactivation. The inactivation mechanism includes the hydroxylation of aminogroups by NO synthase, the formation of nitroso intermediates, and their denitrosation with NO release. PMID- 17086965 TI - [Some problems of studying spatial heterogeneity of animal communities]. AB - The efficiency of different techniques of data collection and processing was considered by the example of bird communities. Accounted variance of the coefficients of bird complex similarity was used as the measure. Annual censuses in different sites proved more expedient than a long-term census in a single key site to study medium- and small-scale spatial heterogeneity of bird communities. Averaging and generalization of data collected in different years and sites yielded satisfactory results. Estimated informativeness in this case proved closer to the real one compared to averaged long-term data since the included representation errors in space and time. PMID- 17086966 TI - [The relationship between the resistance of Aegilops umbellulata Zhuk. seedlings to Septoria nodorum Berk. and peroxidase isozyme pattern]. AB - The resistance of Aegilops umbellulata Zhuk. to blotch pathogen Septoria nodorum Berk. has been studied. Cytoplasmic peroxidase activity in normal seedlings could not be used for biochemical prediction of Ae. umbellulata resistance to Septoria blotch. At the same time, isoperoxidases with pI approximately 7.5 and infection induced activity of anionic isoforms with pI approximately 3.5 were markers of the resistant Aegilops plants. Aegilops resistance to the fungus depended on the activity of cell wall peroxidases. PMID- 17086967 TI - [Hormonal status of tobacco variety Samsun NN exposed to synthetic coenzyme Q10 (ubiquinone 50) and TMV infection]. AB - Hormonal system status has been analyzed in leaf disks of hypersensitive tobacco Nicotiana tabacum L. variety Samsun NN during the development of resistance to tobacco mosaic virus (TMV) induced by synthetic coenzyme Q10 (ubiquinone 50). The absolute and relative content of abscisic acid (ABA), indoleacetic acid (IAA), and cytokinins (CKs) was determined after the exposure of leaves to Q10 solution and the subsequent TMV infection. In plants not treated with Q10, CK content increased about 2.5 times 1 day after TMV infection, while a significant increase in the ABA level and a decrease in the IAA level were observed only after 2 days. In the dynamics, Q10 treatment had a protective antiviral effect, significantly decreased the ABA level, and increased the IAA level in sensitized plants compared to nonsensitized ones. PMID- 17086969 TI - [Water balance and adaptation strategy to extreme climatic conditions in insects of Central Yakutiia]. AB - Original experimental data describing the strategy for cold adaptation of the winter form of black-veined white Aporia crataegi L. living in Central Yakutia, a region with a harsh continental climate and long and very cold winters, are presented in comparison with other insects of Yakutia. Open wintering insects were shown to develop an efficient water-conservation mechanism in a frozen state. This can be significant for their survival in post-diapause. Certain relationships between water balance, wintering mode, and strategy for cold adaptation have been revealed by the example of several insect species of this region in comparison with insects living in less severe climatic conditions. PMID- 17086968 TI - [Variation in sex pheromone composition in large fruit-tree tortrix Archips podana Scop. (Lepidoptera: Tortricidae)]. AB - Variation in sex pheromone composition was studied in large fruit-tree tortrix. The pheromone contained cis-11-tetradecenyl acetate and trans-11-tetradecenyl acetate as macrocomponents as well as microcomponents cis-11-tetradecenol and trans-11-tetradecenol with the prevalence of cis-11-tetradecenyl acetate and cis 11-tetradecenol. The pheromone composition depended in part on the genotype of the male parent. Inbreeding increased the proportion of trans-11-tetradecenyl acetate and trans-11-tetradecenol. PMID- 17086970 TI - [Antiplatelet effect of adenosine triphosphate administration to animals under different experimental conditions]. AB - A significant and considerable decrease in abnormally high platelet aggregation has been demonstrated after intramuscular administration of sodium adenosine triphosphate (ATP) to rats with depressed anti-coagulant system (in aging rats at the age of 11-12 months) and to rats with experimental diabetes both preliminarily and at the background of progressing diabetes. The elimination of one of thrombotic risk factors (decreasing elevated platelet aggregation) points to possible antithrombotic activity of ATP under these experimental conditions. PMID- 17086971 TI - [Effect of Siberian tree species on N2O production and consumption]. AB - The effect of six Siberian tree species on two stages of denitrification-N2O production and consumption-was studied. Broadleaf species (aspen and birch) proved to have lower rates of N2O consumption compared to coniferous species. The factors influencing production and consumption of N2O were also evaluated. The replacement of coniferous forests with broadleaf trees will double the N2O/N2 ratio in the denitrification end-products. Doubled N2O emission from Siberian forest soils to the atmosphere can be expected due to changes in tree species composition of forest ecosystems even without considering changes in water and temperature regimes in soil. PMID- 17086972 TI - [Some rules of soil invertebrate community organization (by the example of Amur Basin mesofauna)]. AB - Some rules of organization of soil communities in the zone of mixed, Jeddo spruce, and larch taiga as well as forest-steppe in the Amur Basin have been formulated on the basis of the factual data. Abundant resources in the soil layer proved to decrease the competition in favor of neutralism, which allows sister species to occupy the same ecological niche. The stability of soil conditions allows considerable contraction of the realized niche volume, which increases species richness in soil. The biocenotic significance of soil organisms is determined by their biomass, productivity, and functional interchangeability. Ecological redundancy makes the mesofauna community much more resistant to environmental factors. The interaction type can change with environmental conditions. Competition prevails in soil communities outside their ecological optimum. Under these conditions, the community demonstrates the concentration of domination and decreased species richness due to the disappearance of closely related invertebrate species largely in biologically regressing groups. PMID- 17086973 TI - [Diazotrophs in the digestive tract of termite Neotermes castaneus]. AB - Normal vital activity of termites Neotermes castaneus requires the presence and continuous replenishment of transient nitrogen-fixing bacteria in their digestive tract, which is realized by coprophagy and repeated utilization of substrate enriched in termite feces. This is the first demonstration of significant changes in the complex of aerobic and facultatively anaerobic nitrogen fixers mediated by an extension of microbial group composition and a shift in dominant taxa in termites that cannot reutilize their feces. PMID- 17086974 TI - [Characterization of crystal-forming bacteria Bacillus thuringiensis subsp. tohokuensis toxic to mosquitos]. AB - Distribution study of Bacillus thuringiensis strains in Western Siberian soils allowed us to isolate crystal-forming bacteria assigned to a new pathovar of Bacillus thuringiensis ssp. tohokuensis with a toxic effect on mosquito larvae. A description of this bacterial pathovar is presented. PMID- 17086975 TI - [Effect of light on white blood cell count in carp Cyprinus carpio L]. AB - In the dark, the lymphocyte count decreased while the monocyte and basophil counts increased in the carp blood. Conversely, the lymphocyte count increased and the monocyte and basophil counts decreased during 12D:12L, 16D:8L, and 0D:24L dark/light cycles, which can be considered as an improvement of the overall physiological state of the yearlings. High light intensity improved the physiological state of carp yearlings, while zero illumination had an inverse effect. No pattern has been revealed for the effect of intermediate dark/light cycles. PMID- 17086976 TI - [The structural changes in the ribosome during the elongation cycle]. AB - The plenty of data about structural changes in the ribosome during its functioning has been accumulated. The most interesting information on such changes was obtained by cryo-EM of various ribosomal complexes with the ligands and by combination of rRNA site-directed mutagenesis with the analysis of structural changes in ribosome by chemical modification technique (chemical probing). The most studied structural transformations of the ribosome interacting with tRNAs and elongation factors are considered in this review. The structural rearrangements are discussed in the context of interactions between the functional centers of the ribosome. We also describe the system of tertiary contacts between the rRNA helices and proteins which forms the universal structure in the ribosome. We pay attention that by means of such system the allosteric conformational signal can be transmitted between the functional centers. Besides the discussion of different biochemical data in the scope of structural data we also consider the hypothesis that the position of GTPase associated center (GAC) in the ribosome regulates the binding of elongation factors. PMID- 17086977 TI - [GTPases of prokaryotic translational apparatus]. AB - Four protein factors, belonging to the GTPase superfamily, participate in bacterial biosynthesis: IF2, EF-G, EF-Tu and RF3. The exact role and mechanism of action of these proteins was of particular interest over the last several decades. Recent advances in structural methods of ribosomal research, especially application of cryoelectron microscopy, provided powerful experimental tools for the investigation of ribosomal dynamics during translation. Simultaneously, progress in the biochemical investigation of translation allowed us to link structural rearrangements occurring in the ribosome to functional changes in the ribosome-bound translational GTPases--GDP/GTP exchange, GTPase activation and its conformational changes. Accumulated data have lead to formulation of current models of mechanisms of translation. More and more facts testify in favor of the idea that the ribosome plays a prominent role both in the nucleotide exchange and in GTPase activation, thus playing the role both of GAP and GEF for RF3, IF2 and EF-G. In our work we attempted to systematize the most important experimental findings and models for mechanisms of GTPases function and regulation in prokaryotic translation. PMID- 17086978 TI - [Gene silencing of HIV-1 using genetic constructs expressing siRNAS]. PMID- 17086979 TI - [Alternative translation start sites and their significance for eukaryotic proteome]. AB - Current models of translation initiation and contextual organization of eukaryotic mRNA leader region are reviewed. Hypothesis on frequent usage of several alternative start codons is discussed. Potential contribution of alternative translation start sites to eukaryotic mRNA coding potential is considered. PMID- 17086980 TI - [Biochemical and molecular characterization of gliadins]. AB - Gliadins account for about 40-50% of the total proteins in wheat seeds and play an important role on the nutritional and processing quality of flour. Usually, gliadins could be divided into alpha- (alpha/beta-), gamma- and omega-groups, whereas the low-molecular-weigh (LMW) gliadins were novel seed storage proteins. The low-molecular-weight glutenin subunits (LMW-GSs) were also designated as gliadins in a few literatures. The genes encoding gliadins were mainly located on the short arms of group 6 and group 1 chromosomes, and not evenly distributed. Repetitive sequences covered most of un-coding regions, which attributed greatly to the evolution of wheat genome. Primary structure of each gliadin has been divided into several domains, and the long repetitive domains consisted of peptide motifs. Conserved cysteine residues mainly formed intramolecular disulphide bonds. The rare potential intermolecular disulphide bonds and the long repetitive domains played an important role in the wheat flour quality. There was a general idea that gliadin genes, even prolamin genes, have a common origin and subsequent divergence lead to the gene polymorphism. The gamma-gliadins have been considered to be the most ancient of the wheat prolamin family. Several elements in the 5'-flanking (e.g. CAAT and TATA box) and the 3'-flanking sequences had been detected, which had been shown necessary for the proper expression of gliadins. PMID- 17086981 TI - [Identification of a novel human MAST4 gene, a new member of the microtubule associated serine-threonine kinase family]. AB - Human protein kinases make up a large superfamily of homologous proteins, which are related by virtue of their kinase domains (also known as catalytic domains). Here we report the cloning and characterization of a novel human MAST4 (microtubule associated serine/threonine kinase family member 4) gene, which locates on human chromosome 5q13. The MAST4 cDNA is 7587 base pairs in length and encodes a putative protein of 2435 amino acids which contains a serine/threonine kinase domain and a PDZ domain. MAST4 protein has 64%, 63%, 59% and 39% identical aminoacid residues with MAST1, MAST2, MAST3 and MASTL respectively. RT-PCR analysis revealed relatively high expression level of MAST4 in most normal human tissues, with an exception of in testis, small intestine, colon and peripheral blood leukocyte. PMID- 17086982 TI - [Molecular cloning and characterization of a novel WRKY gene from Brassica chinensis]. AB - A new WRKY gene was cloned from Brassica chinensis by rapid amplification of cDNA ends (RACE). The full-length cDNA of BcWRKY was 1175 bp long and contained a 924 bp open reading frame (ORF) encoding a putative W-box-binding protein of 308 amino acids. The predicted BcWRKY protein was found to have a potential bipartite nuclear localization sequence (NLS-BP) in its N-terminal region followed by a WRKY DNA-binding domain. Bioinformatic analysis revealed that BcWRKY resembled other WRKY domain-containing proteins from Arabidopsis (AtWRKY18), tobacco (WIZZ), parsley (PcWRKY4) and wild oat (ABF2). Expression of the BcWRKY gene could be induced by salicylic acid (SA), and influenced by Pseudomonas syringae pv. tomato strain DC3000 infection and wounding treatment. Our study implies that BcWRKY might have similar functions possessed by other WRKY genes such as inducing the expression of some defense-related genes and increasing plant's disease resistance ability. PMID- 17086983 TI - [Isolation by suppression-subtractive hybridization of genes preferentially expressed during early and late fiber development stages in cotton]. AB - As a main natural fiber source, cotton plays an important role in human life. To identify genes preferentially expressed during early and late cotton fiber development, we constructed two fiber subtracted libraries on the basis of PCR selected subtraction using a pool of nonfiber tissues as the same driver and 10 days postanthesis (DPA) and 20 DPA fiber cells as testers, respectively. Through differential screening, 292 clones in both libraries were identified as being preferentially expressed during fiber development. Sequence analysis showed that 31 unique sequences were found in the library of 10 DPA fibers and 48 unique sequences were obtained in the library of 20 DPA fibers. In addition, there were 13 unique clones in common in both libraries. Many previously reported cotton fiber-related genes were included in both libraries. Northern hybridization was performed to further confirm the differential expression and to determine the pattern of mRNA accumulation of selected clones. As a result, all selected cDNAs showed highly preferential expression in developing cotton fibers. Four genes putative gibberellin-regulated protein, putative tonoplast intrinsic protein, putative plasma membrane intrinsic protein, and Gossypium hirsutum putative membrane protein-were identified in 10 DPA fiber subtracted library, and they were found to be expressed a lot during early fiber development. On the other hand, those genes screened out of 20 DPA fiber subtracted library, like arabinogalactan protein and fiber glycosyl hydrolase family 19 protein, were found highly expressed in fibers with the maximal transcription level during developmental switch from elongation to cellulose deposition. By subtraction between fibers and five nonfiber tissues, two sets of genes were identified, and their fiber-specific or fiber-preferential expression indicated that they are involved in the network that controls cotton fiber development. PMID- 17086984 TI - [Nah-plasmids of IncP-9 group from natural strains of Pseudomonas]. AB - Use of polymerase chain reaction helped to establish that the most frequent among naphthalene utilizing bacteria, isolated on the territory of Belarus, are Nah plasmids of IncP-9 incompatibility group and those with indefinite systematic belonging. With the help of classical test of incompatibility, restriction and sequence analyses three new subgroups within the IncP-9 group were discovered (zeta, eta and IncP-9-like replicons). Conducting of restriction analysis for amplification products of nahG and nahAc genes allowed us to reveal, in addition to known sequences of stated determinants, two new types of nahG gene. Restriction analysis performed on amplification products of 16S RNA genes (ARDRA method) showed that native hosts of Nah-plasmids of IncP-9 group are not only fluorescent bacteria from genus Pseudomonas (P. fluorescens, P. putida, P. aeruginosa, P. species), but also non-fluorescent bacteria with indefinite specific belonging. PMID- 17086985 TI - [Phenotypic manifestation of epigenetic determinant [ISP+] in Saccharomyces serevisiae depends on combination of mutations in SUP35 and SUP45 genes]. AB - It is known that translation fidelity in Saccharomyces yeast is determined by factors of genetic and epigenetic (prion) nature. The work represents results of further analysis of strains containing non-chromosomal determinant [ISP+], described earlier. This determinant is involved in the control of translation fidelity and some of its properties indicate that it is a prion. [ISP+] manifests phenotypically as antisuppressor of two sup35 mutations and can be cured by guanidine hydrochloride (GuHCl). Here we have shown that sup35 mutants containing [ISP+] contain also additional sup45 mutations. These mutations cause amino acid replacements in different regions of eRF1 translation termination factor, encoded by SUP45 gene. Strains bearing sup35-25 mutation contain sup45 mutation, which causes amino acid replacement at position 400 of eRF1; strains bearing sup35-10 contain mutation causing replacement, which alters eRF1 at position 75. Thus, antisuppressor phenotype of [ISP+] strains depends on interaction of sup35 and sup45 mutations, as well as on the GuHCl-curable epigenetic determinant. PMID- 17086986 TI - [Mutation analysis of functional role of amino acid residues in domain IV of elongation factor G]. AB - Seven variants of elongation factor G (EF-G) from Thermus thermophilus with mutations Glu494Ile, Gly495Asp, Lys496Ile, His509Leu, Lys564Ile and Tyr568Lys located in the beta-sheet of its domain IV and mutation Gly553Asp in a loop between domain III and IV were constructed using polymerase chain reaction. Functional tests demonstrated that only mutation Lys496Ile, located in the vicinity of the loop 501-504, inhibits translocation effectiveness, in the presence of the mutated EF-G. The functional analysis of all mutations constructed up to now in domain IV reveals that only those located in loops 501 504 and 573-578 markedly decrease the translocation activity of EF-G. These loops are located at the tip of domain IV and close to the decoding center of the 30S ribosomal subunit upon EF-G interaction with the ribosome. The functional role of EF-G and its domain IV in ribosomal translocation is discussed. PMID- 17086987 TI - [DNA-hydrolyzing IgG antibodies from the blood of patients with acquired immune deficiency syndrome]. AB - DNAase activity of 110 samples of IgG from the blood of AIDS patients was analyzed. It was shown that the relative activity of preparations varies very much from patient to patient, but 96% preparations show detectable level of DNAase activity. Several rigid criteria were applied and it was shown that DNAase activity is an intrinsic property of antibodies from AIDS patients. It was shown that catalytic activity could posses not only intact IgG, but also separated light chains of polyclonal antibodies. The abzymes catalyze DNA hydrolysis effectively in a wild range of pH (5.0-9.5). K(M) and V(MaKC) values of antibody dependent hydrolysis of DNA was estimated. PMID- 17086988 TI - [Activation of RHOA gene transcription in epithelial tumors may be caused by gene amplification and/or demethylation of its promotor region]. AB - RHOA protein, a member of small GTPases family, is implicated in cell morphogenesis, adhesion, and in cell cycle regulation. RHOA gene (3p21.31) exhibits cell transformation activity, and therefore gene is considered as a potential oncogene. The aim of this study was to investigate RHOA transcription and copy number changes in three epithelial tumors (breast, renal cell and epithelial ovarian carcinomas, 45 tumor/normal pairs altogether). EII, HhaI, AciI n Bsh1236I). Hypomethylation of the RHOA promoter region in tumor DNA was observed two times more frequently than increased methylation. Moreover, all (15) cancer cases with hypomethylation of the RHOA gene showed a 2-10 fold increased expression of RHOA. It was concluded that gene copy multiplication and demethylation of the RHOA promoter region can contribute to transcription activation of this gene in epithelial tumors. PMID- 17086989 TI - [Illegitimate recombination as a possible mechanism of DNA-topoisomerase II induced chromosomal rearrangements]. AB - Using semi-quantitative PCR-based approach, we have shown that the breakpoint cluster region of the AML1 gene was associated with the nuclear matrix. We have demonstrated that inhibition of topoisomerase II by etoposide stimulates the appearance of histone gammaH2AX foci, an indicator for the presence of DNA double strand breaks. Furthermore, the major part of these foci was associated with the nuclear matrix. We also visualized nuclear matrix--associated multiprotein complexes involved in topoisomerase II--induced DNA double-strand break repair. Colocalization studies have demonstrated that these complexes included the principal components of the non-homologous end joining repair system (Ku80, DNA PKcs and DNA ligase IV). Thus, it is reasonable to suggest that the non homologous DNA end joining is a possible mechanism of topoisomerase II--induced chromosomal rearrangements. PMID- 17086990 TI - [The short form of heparin-binding EGF-like growth factor. Part II]. AB - The structure of monkey (Chlorocebus aethiops) heparin-binding EGF-like growth factor (HB-EGF) gene has been investigated in this work in comparison with the known structure of human gene. It was shown that HB-EGF short form (SF-HB-EGF) specific exon 3a is mapped between exons 3 and 4 at distance 700 b.p. from exon 4. In a number of human and simian cell lines the main part of SF-HB-EGF mRNA does not contain HB-EGF mRNA specific exons 4 and 5. In comparison with HB-EGF mRNA in SF-HB-EGF mRNA P-form, but not L-form of is predominant, and this mRNA encodes a polypeptide with changed propeptide structure. Labeled SF-HB-EGF competes with HB-EGF and EGF for binding sites at A431 cell surface, which may be due to interaction with specific receptor. All the data suggest a specific role of SF-HB-EGF in cellular signalization. PMID- 17086991 TI - [1H NMR study of the complexation of the quinolone antibiotic norfloxacin with DNA]. AB - Complexation of antibiotic norfloxacin (NOR) with DNA fragments 5'-d(TpGpCpA) and 5'-d(CpGpCpG) has been studied in aqueous solution by 1H NMR spectroscopy (500 MHz). Equilibrium parameters of the complexation with single-stranded and duplex forms of DNA oligomer--equilibrium constants, enthalpy and entropy--have been obtained for the first time. Based on the analysis of the complexation parameters as well as induced chemical shifts of the antibiotic protons within different complexes, it was found that NOR binds with the tetramer duplexes mainly by intercalation. The complexation with the single-stranded form may occur either by intercalation and external binding. The site of preferential binding of the antibiotic with DNA duplex is GC site. PMID- 17086992 TI - [The conformational dynamics of the tetramer hemoglobin molecule as revealed by hydrogen exchange. III. Influence of the heme removal]. AB - Two main types of conformational fluctuations--local and global are characteristic of the native protein structure and revealed by hydrogen exchange. The probability of those fluctuations changes to a different extent upon hemoglobin oxygenation, changing of pH, splitting of the intersubunit contacts. To compare with the influence of the heme removal the rate of the H-D exchange of the peptide NH atoms of the human apoHb was studied at the pH range 5.5-9.0 and temperature 10-38 degrees C by the IR spectroscopy. The removal of the heme increases the rate of the H-D exchange of the 80% peptide NH atoms with the factor retardation of the exchange rate (P) in the range approximately 10(2) 10(8). For the most of the peptide NH atoms the probability of the local fluctuations weakly depends on the temperature, the enthalpy changes upon all such local conformational transitions deltaH(op) degrees are 0-15 kcal/M. Characterized by the stronger temperature dependence the global fluctuations are not arised upon the temperature increases up to 38 degrees C at pH 7.0 inspite of in these conditions the slow denaturation and aggregation of apoHb begin to occur. Upon the destabilization of the apoHb structure by the simultaneous decreasing of pH to 5.5 and temperature to 10 degrees C the global fluctuations of the apoHb native structure described by deltaH(op)o < 0 begin to intensify. The mechanism of the overall intensification of the local fluctuations upon the heme removal, the peculiarity of the heat denaturation of apoHb in conditions, close to that existing upon the selfassembly of Hb in vivo, and analogy between low temperature global fluctuations and cold denaturation of globular proteins are discussed. PMID- 17086993 TI - [A search for amyloidogenic regions in protein chain]. AB - We suggest a new method to detect amyloidogenic regions in a protein sequence. In the present work it is shown that regions enriched with amino acid residues which have a high expected packing density are responsible for the amyloid formation. Our predictions are consistent with known disease-related amyloidogenic regions for 8 of 11 amyloid-forming proteins and peptides in which positions of amyloidogenic regions have been revealed experimentally. PMID- 17086994 TI - [Regulation of nitrogen metabolism in gram-positive bacteria]. AB - We searched for new members of the TnrA and GlnR regulons controlling assimilation of nitrogen in gram-positive bacteria. We identified the regulatory signals for these transcription factors with consensuses ATGTNAWWWWWWWTNACAT for GlnR and TGTNAWWWWWWWTNACA for TnrA. We described the structure and found new potential members for the TnrA/GlnR regulons in Bacillus subtilis, B. licheniformis, Geobacillus kaustophilus, Oceanobacillus iheyensis, for the TnrA regulon in B. halodurans and for the GlnR regulons in Lactococcus lactis, Lactobacillus plantarum, Streptococcus pyogenes, S. pneumoniae, S. mutans, S. agalactiae, Enterococcus faecalis, Listeria monocytogenes, Staphylococcus aureus and St. epidermidis. PMID- 17086995 TI - Epidemiology of HIV infection in Lebanon. Data from 1985-2005. AB - Lebanon is considered among the countries where the HIV epidemic is still in its early phase of spread. Little information is available on the magnitude of the problem due to the lack of an efficient surveillance system. In this review, we report on the epidemiology of HIV infection in Lebanon, based on limited data generated by the Lebanese National AIDS Control Program (NAP). By May 2005, a cumulative number of 813 cases had been diagnosed. The most prevalent mode of HIV transmission in Lebanon is heterosexual exposure accounting for around 50% of the cases. Two hundred ninety-six patients (36.4%) fulfilled the AIDS case definition. Despite the availability of highly active antiretroviral therapy (HAART) provided by the Ministry of Health, the disease burden of HIV infection remains unknown in this country. PMID- 17086996 TI - Primary care of HIV-infected adults. AB - HIV infection is increasingly becoming a chronic illness as effective treatment allows patients to live longer and stay healthier. HIV-infected patients often benefit from having both a generalist familiar with primary care medicine and a HIV expert involved in their care. This article is geared at the generalist who is involved with the care of HIV-infected patients and presents a series of cases to address topics important in the primary care of patients with HIV infection. PMID- 17086997 TI - Antiretroviral therapy in HIV. AB - The advent of highly active antiretroviral therapy (HAART) has led to a significant decline in HIV-associated morbidity and mortality. Despite these recent advances, the majority of patients with HIV/AIDS worldwide are still lacking access to treatment. To date, there are 21 approved antiretrovirals divided into four categories based on their mechanism of action. We review the basic characteristics of these antiretrovirals. We also briefly discuss current guidelines regarding the use of antiretrovirals in general as well as in special populations such as in pregnant women, in occupational exposure and non occupational exposure. PMID- 17086998 TI - HIV-associated opportunistic infections. Bacterial infections. AB - Bacterial infections are the most common opportunistic infections in HIV. Since the advent of highly active antiretroviral therapy (HAART), the incidence of these infections is on the decline. However, globally there is a significant lack of access to care among HIV patients because of limited drug availability and cost of treatment. Furthermore, non-compliance and drug resistance can hinder viral suppression, predisposing patients to opportunistic infections. We review the major bacterial opportunistic infections in HIV positive patients including tuberculosis, Mycobacterium avium complex infections, syphilis, bacterial enteric disease, bacterial pneumonia and bartonellosis. Epidemiology, clinical presentation, diagnosis, treatment and prophylaxis are also reviewed. PMID- 17086999 TI - Opportunistic infections in patients with HIV and AIDS. Fungal and parasitic infections. AB - Since the advent of highly active antiretroviral therapy (HAART), the incidence of specific fungal and parasitic opportunistic infections as observed in HIV disease has been in decline. However, in many parts of the world, and in areas where access to HAART is limited, these specific opportunistic infections are still observed in HIV patients. In this manuscript, we review the epidemiology, clinical manifestations, diagnosis, and treatment of common fungal and parasitic opportunistic infections. PMID- 17087000 TI - Viral opportunistic infections in HIV-infected adults. AB - Despite the development of highly active antiretroviral therapy (HAART), opportunistic infections continue to be seen in HIV-infected patients throughout the world. The primary reason for this is the lack of access to HAART for most people living with HIV/AIDS. For patients that have access to HAART, some may not have an effective response to therapy, due to reasons such as medication toxicity, poor adherence, or drug-resistant strains of HIV. Viral infections, in particular, are a major cause of opportunistic infections in HIV-infected adults, and can lead to significant morbidity and mortality. We have reviewed the epidemiology, clinical manifestations, diagnosis, and treatment of the most common viral opportunistic infections, including cytomegalovirus, JC virus, varicella-zoster virus, herpes simplex virus, and human papillomavirus. PMID- 17087001 TI - Metabolic and morphologic complications of HIV infection. AB - In the era of highly active antiretroviral therapy, long-term complications of HIV infection and antiretroviral therapy deserve heightened attention. Morphologic and metabolic complications seen during the course of HIV infection encompass a variety of entities that may share a common pathophysiologic pathway. This review article will discuss clinical syndromes such as wasting, lipoatrophy/lipohypertrophy, polymetabolic syndrome as well as hyperlipidemia, cardiovascular disease, lactic acidosis, and metabolic bone disease in HIV infected patients. PMID- 17087002 TI - Human immunodeficiency virus and hepatitis C virus co-infection. AB - Hepatitis C virus (HCV) infection is a major cause of liver disease and hepatocellular carcinoma worldwide, as well as the leading cause of liver transplantations in the United States. As a result of similar modes of transmission, approximately 30% of HIV-infected individuals are co-infected with HCV. Among intravenous drug users, almost 90% of people infected with HIV are also infected with HCV. Because of treatment with highly active anti-retroviral therapy, HIV-infected individuals have improved survival and are no longer suffering from opportunistic infections and malignancy as in years past. As a result, co-infection with HCV has now become a frequent cause of morbidity and mortality in HIV-infected individuals. Furthermore, liver disease secondary to HCV infection is now the leading cause of hospital deaths in HIV-infected people in the US. HIV infection accelerates the course of HCV-related liver disease and viremia. It is less clear whether HCV infection affects the clinical course of HIV; however, HCV-related liver disease can limit many individuals from receiving anti-HIV therapy. HIV/ HCV co-infection is common, and serious. Physicians caring for HIV-infected patients worldwide must now address hepatitis C virus co infection. PMID- 17087003 TI - HIV-associated malignancies. PMID- 17087004 TI - [Evolution biomedicine is a new trend in biological science]. PMID- 17087005 TI - [Study of cholinesterases from different species using an inhibition analysis technique (structure variation of dialkoxyphosphate phosphoryl and cleaved groups)]. PMID- 17087006 TI - [Structural and functional organisation of G protein signal systems of amoeba Dictyostelium discoideum ]. PMID- 17087007 TI - [Structural changes in the erythrocyte membrane of the rainbow trout Salmo irideus in seasonal acclimatization]. PMID- 17087008 TI - [Comparison of malate dehydrogenase isozymes from the hare and rabbit heart]. PMID- 17087009 TI - [Carnosine and relative compounds protect double-stranded DNA from oxidative damage]. PMID- 17087010 TI - [Adaptation of the nematode Caenorhabditis elegans to a high temperature of the environment]. PMID- 17087011 TI - [Effect of antagonists of inhibiting amino acids on motoneuron postsynaptic potentials in Rana ridibunda]. PMID- 17087012 TI - [Trophic relationship in the system host (Lota lota) - parasite (Eubothrium rugosum): symbiont microflora in hydrolysis of food carbohydrate components]. PMID- 17087013 TI - [Processing of locally meaningful spectral changes in sound signals by the neurons of corpora quadrigemina and auditory cortex in Mus musculus]. PMID- 17087014 TI - [Study of intellectual activity during recognising multitude of two- and three dimension stimuli of different colors in lower monkeys]. PMID- 17087015 TI - [Organisation of the midbrain in the sturgeon. Dorsothalamus and epithalamus]. PMID- 17087016 TI - [Intrauterine infections]. AB - A broad variety of microorganisms are capable of causing fetal infections. Among viral agents prominent are the human cytomegaly virus (HCMV), herpes simplex virus (HSV), human immundeficiency virus (HIV), varicella, herpes zooster, rubella, parvovirus B19, measles and the hepatitis B and C viruses. Protozoa such as toxoplasma and spirocheta pallida, causing congenital syphilis are equally important. Bacterial infections are responsible for in uterus aquired listeriosis, tuberculosis, and group B streptococcus infections. Fungi including candida albicans complete the circle of infections pathogens. Infectious microrganisms may reach the fetus through the placenta are may ascend through the birth canal. The quoted pathological agents threaten the health and life of the fetus directly by the biological derangements they cause and also by inducing abortion or premature birth. The clinical manifestations include retarded growth, central nervous system damage and skin lesions. The invariable therapeutic measures vary but in general, are limited value in cases of in utero acquired infections. PMID- 17087017 TI - [The function of the immune system after the treatment of pediatric malignant diseases]. AB - INTRODUCTION: Currently, malignancies in childhood can be cured in 70 percent of the cases. However, the intensive cytostatic therapy may lead to late side effects influencing quality of life. AIM OF THE STUDY: Analysis of the reconvalescence of the immune functions after completion of therapy for malignancies in children. PATIENTS: 88 long-term survivors (51 boys, 37 girls) were investigated (43 acute lymphoid leukemia, 15 lymphoma, 20 bone tumors, 10 other solid tumors). Mean age at the time of diagnosis was 7.8 years (1 mo-17.7 years). METHODS: The following parameters were investigated: serum immunoglobulin levels after completion of the chemotherapy and in the next 4 years thereafter, lymphocyte subpopulations in the peripheral blood by flow-cytometry and cellular immunity by in vitro tests (natural killer activity, antibody-dependent cellular cytotoxicity, mitogen-induced T- and B-cell blastic transformations). RESULTS: Lower serum immunoglobulin (IgG) levels could be detected in patients with leukemia after completion of the chemotherapy (8.8 +/- 3.2 g/l). One year thereafter serum IgG levels increased significantly (10.1 +/- 2.9 g/l) (p<0.05). In patients with solid tumors the serum IgG levels were in the normal range at the end of the chemotherapy (12.1 +/- 4.3 g/l). At a mean of 1.3 years after the end of chemotherapy NK activity decreased in 7/43 (16.3%) leukemia patients, and in 3/45 (6.7%) solid tumor patients, ADCC decreased in 8/43 (18.6%) and 3/45 (6.7%), respectively (p<0.05 leukemia vs. solid tumor). At a mean of 15 months after the end of the therapy B-cell blastic transformation was decreased in 3/43 (7%) leukemia patients and in 4/45 (8.9%) solid tumor patients. At the same time point T-cell blastic transformation was altered in 5/43 (11.6%) and in 4/45 (8.9%) cases, respectively. CONCLUSION: Cytotoxic therapies lead to severe, long term depression of the immune system. At the end of the chemotherapy this effect is more pronounced in leukemia patients. Years (1.5-3) after completion of the therapy in a significant proportion of the patients some in vitro parameters of the immune system are yet altered, so careful monitoring of this patient population is mandatory. PMID- 17087018 TI - [First Hungarian report of olopatadine eyedrop therapy in children and adults suffering from seasonal allergic conjunctivitis]. AB - The eye drop Opatanol (its main component is olopatadine, which has an antihistaminic and membrane-stabilizing effect) in Hungary is not well known yet. PATIENTS AND METHODS: In 2004 and 2005 in the ragweed season we had treat with olopatadine eyedrop for 2 weeks (two times daily) 37 children, who had ragweed allergy. The average age of the 23 boys was 8.8 years, and for the 14 girls 10.5 years. In July and August of 2004 10 adult allergic female (average of their age: 29,2 years) also had receive olopatadine for 2 weeks. The severity of the symptoms was recorded by using a scale from 0 to 3 (where 3 indicates the most severe symptoms) before and 2 weeks after the beginning of the treatment, and the average was calculated. RESULTS: The discomfort feeling decreased from 2.4 +/- 0.5 to 0.8 +/- 0.2 in children, and from 1.5 +/- 0.9 to 0 in adults, the itching from 2.5 +/- 0.4 to 0.2 +/- 0.1 in children, and from 1.6 +/- 1.2 to 0 in adults, the foreign body sensation from 1.6 +/- 0.9 to 0.7 +/- 0.4 in children, and from 1.3 +/- 1.1 to 0 in adults, the tearing from 1.9 +/- 0.9 to 1.0 +/- 0.3 in children, and from 1.0 +/- 1.0 to 0.1 in adults, and the redness from 2.2 +/- 0.4 to 1.1 +/- 0.2 in children and from 1.9 +/- 0.8 to 0.5 +/- 0.5 in adults. We observed side effect in one child (the redness of the conjunctiva increased after the application of the eye drop) and in two adults (burning sensation for short time, in the other case foreign body sensation for 30 seconds). CONCLUSIONS: The anti-allergic eye drop olopatadine is a safe and effective treatment for seasonal allergic conjunctivitis even in childhood. It is comfortable, to use twice daily is sufficient. PMID- 17087019 TI - [Haptoglobin polymorphism in patients with inflammatory bowel diseases]. AB - BACKGROUND: Since functional differences were found among three major haptoglobin phenotypes, haptoglobin polymorphism was reported to be associated with the risk and clinical course of different inflammatory diseases. The aim of the study was to investigate the Hp polymorphism distribution in Hungarian Crohn's disease patients. METHODS: 511 Hungarian IBD patients were investigated (Crohn's disease patients: 468, m/f ratio: 233/235, duration 8.2 +/- 6.7 ys, and ulcerative colitis patients: 43, m/f: 22/21, duration: 9.5 +/- 10.6 ys) and 384 healthy subjects served as controls. Hp phenotypes were determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis of sera followed by immunoblotting. Clinical data were come by the questionnaires prepared by the physicians. RESULTS: The frequency of haptoglobin-1 allele was significantly higher in Crohn's disease (0.395) compared to the controls (0.345; OR: 1.24, 95%CI: 1.02 1.52, p = 0.03), but the phenotype distribution showed no such differences. Haptoglobin phenotype was associated to disease behavior in Crohn's disease (B1 and B2, in haptoglobin 1-1 and 2-2: 36.6%-34.3% and 32.4%-32.5% vs. in 2-1: 44.9% and 20.3%; ORB1Hp2-1 vs. others: 2.06, 95%CI: 1.29-3.28). Furthermore, an increased frequency of primary sclerosing cholangitis was observed in haptoglobin 2-2, compared to the 1-1 (6.5% vs. 0.0%, p = 0.039). No associations were found in ulcerative colitis. CONCLUSIONS: haptoglobin-1 allele was associated with Crohn's disease, whereas the phenotypes with the disease behavior and frequency of primary sclerosing cholangitis, exhibiting a disease-modifying effect. PMID- 17087020 TI - [Prof. Ozsvath Karoly is 80 years old]. PMID- 17087021 TI - [100 years of the Hungarian Surgeon Society]. PMID- 17087022 TI - [In memoriam Zoltan Rosenberg (1927-1990)]. PMID- 17087023 TI - [Value of color doppler imaging in the diagnosis of occlusive retinal venous lesions]. AB - A hundred and twenty-five patients aged 47 to 72 years who had retinal venous occlusions were examined. Occlusion of the central retinal vein (CRV) was observed in 82 (65.6%) patients and that of CRV branches was seen in 43 (34.4%) patients. In all the patients, color Doppler imaging (CDI) was performed to study blood flow in the ophthalmic artery, central retinal artery (CRA), CRV, and short posterior ciliary arteries. The study indicated a marked reduction in diastolic velocity, no diastolic component of the Doppler blood flow spectrum, and a significant increase in the CRA resistance index (from 0.9 to 1.0), and a pronounced decrease in the maximum systolic velocity in the CRV, as well as a considerable reduction in blood flow velocity parameters, and a rise in the resistance index in the ophthalmic artery and short posterior ciliary arteries. CDI is a highly informative, non-invasive technique that makes it possible to study the blood flow in the CRA and CRV for the evaluation the severity of disease and the efficiency of therapy for occlusive retinal venous lesions. PMID- 17087024 TI - [Choroidal angioarchitectonics and colorimetric scanning]. AB - The procedure proposed by the authors for colorimetric layer scanning of the fundus of the eye is able not only to identify choroidal capillaries as a uniform system, but also to enhance the accuracy of diagnosis and to increase the volume of findings, by analyzing the digital images of the structures of the eye grounds. A follow-up monitoring of the choroid and retinal pigment epithelium in patients with high-grade myopathy makes it possible to establish a diagnosis of early abnormal changes, to prescribe an appropriate treatment, and to evaluate its efficiency. PMID- 17087025 TI - [Morphological features of the mucosa of the nasal cavity and lacrimal sac in lacrimal passage obstruction]. AB - A comprehensive morphological study of the mucosa of the nasal cavity and lacrimal sac was conducted, by using 160 biopsy specimens taken from 44 patients at endoscopic endonasal dacryocystorhinostomy. In all cases, different areas of the nasal cavity exhibited abnormal changes that were generalized and appeared as inflammatory, dystrophic, and sclerotic processes. PMID- 17087026 TI - [A new method for microdrainage using a porous titanium nickelide endoprosthesis in resurgery of uncompensated glaucoma]. AB - Experimental (23 rabbits) and clinical (42 patients with operated glaucoma) studies were conducted to evaluate the effect of a titanium nickelide implant on ocular tissue in modified deep sclerectomy in order to normalize intraocular pressure. Scleral morphological studies in the area of implant placement revealed the formation of a capsule with "fissures" and vessels. Clinically, there was intraocular pressure compensation in 92.8% of the patients operated on. PMID- 17087027 TI - [Morphological rationale for autografting of musculus rectus ocularis fibers at revascular optic nerve decompression surgery]. AB - The authors made a morphological study of the early proposed and clinically tested revascular optic nerve decompression operation developed to improve blood supply to the initial portion of the optic nerve in glaucoma-induced optic neuropathy. An experiment was carried out on 11 rabbits. The operated and control eyes were enucleated 1, 4, and 6 months after surgery. The experiment has established that muscle autografting into the retroscleral and paraneural space of the optic nerve is followed by a number of cellular reactions with the active participation of satellite cells, some of which may be associated with regenerative processes and presumably regarded as one of the cell autotherapy modalities. PMID- 17087028 TI - [The features of the clinical course of different forms of the dry eye syndrome are the basis for development of adequate treatment modalities]. AB - The paper presents two classifications of the dry eye syndrome: the Madrid and Russian classifications. The creation of its classification is required for the elaboration of treatment policy for this category of patients. The classifications are based on the scientific data on the epidemiology, etiology, and pathogenesis of the dry eye syndrome. Each classification identifies different forms of the dry eye. The classifications virtually published at the same time do not greatly differ from each other and they reflect the current approach to systematizing different forms of the dry eye. The paper analyzes the Madrid versus Russian classifications. PMID- 17087029 TI - [Effects of decreased nystagmus amplitude in children periodically exposed to light]. AB - The effect of steady-state nystagmus suppression has been found upon periodic light exposure of the eye. The paper describes the exposure facility and procedure based on the changes in the amplitude and frequency of light pulse until there is a response of a patient's nystagmus parameters that are continuously monitored by video-assisted computer system. After periodic light exposures, there was a persistent suppression of nystagmus and a steady-state increase in visual acuity by an average of 0.1. PMID- 17087030 TI - [Effect of paraaminobenzoic acid on interleukine-6 production in patients with herpetic keratitis]. AB - The purpose of the study was to compare the effect of para-aminobenzoic acid (PABA) (actipol) on tear interleukin-6 (IL-6) levels in patients with herpetic keratitis and in peripheral blood cells of volunteers in vitro. Enzyme immunoassay was used to measure lacrimal fluid IL-6 levels in the actipol and acyclovir groups before and 1, 2, 3, 4, and 7-8 days after therapy and after clinical recovery in 30 patients with herpetic keratitis. A control group comprised apparently healthy volunteers (n=13, 26 eyes) who used actipol instillations into the conjunctival sac 4-5 times a day. The spontaneous production of cytokines and their production in response to the induction with different PABA concentrations (0.001, 0.01, 0.1, and 10 microkg/ml) were studied on peripheral blood cells taken from 9 volunteers. In the patients with herpetic, actipol was found to reduce and normalize tear IL-6 levels while acyclovir failed to produce this effect. In the healthy individuals, actipol did not affect IL-6 production in the anterior segment of the eye. All study PABA concentrations exerted a modulating effect on the production of IL-6 in the peripheral blood cells in vitro. PMID- 17087031 TI - [Deep layer posterior keratoplasty: preliminary data]. AB - The paper presents preliminary results of deep layer keratoplasty first performed in Russian ophthalmology. It describes its surgical techniques. Deep layer posterior keratoplasty was made in 3 patients with epithelial-and-endothelial dystrophy of the cornea. The operation was simultaneously made with ultrasound cataract phacoemulsification and intraocular lens (IOL) implantation artificial in one case. In the early postoperative period, the fixed position of a transparent keratograft was achieved and the epithelial layer also recovered without signs of bullous keratopathy. The immediate results of posterior keratoplasty inspire optimism. Deep layer keratoplasty is technically more difficult than penetrating keratoplasty, more painstaking and more prolonged. At the same time it has no shortcomings typical of penetrating keratoplasty. The procedure of deep layer keratoplasty requires further long-term study, simplification of surgical techniques, detection of possible complications, and modes of their correction. PMID- 17087032 TI - [Study of risk factors of laser irradiation-induced intraocular lens damage]. AB - The rate of distant damages to intraocular lens (IOL) was studied in relation to its material and manufacture procedure (punching or turning), and its posterior lens capsular position in 120 patients (125 eyes) who had undergone laser secondary cataract discussion. The resistance of lenses made from silicone, polymethylmethacrylate (PMMA), and acryl to YAG laser irradiation (1.076 microm) was studied in an experiment. The findings suggest that the risk for distant IOL damaged in YAG-capsulotomy is higher in PMMA lenses made by punching and silicone IOL. Artificial acryl lenses show a high resistance to YAG laser irradiation. The nature and depth of IOL damages are not entirely determined by the physical properties of a lens material and they are also associated with the distance of a lens and an irradiated structure. If there are specific posterior capsular changes as jelly-like thickening and if convex-designed lenses are implanted, then there is no safe distance between the capsule and IOL, which permanently leads to its damage by YAG-laser irradiation. PMID- 17087033 TI - [Topography and functional changes of the rabbit retina with a single injection of triamcinolone]. AB - Intravitreal injections of 6 and 8 mg of triamcinolone (kenalog) were studied for impact on the time course of electroretinographic changes and the retinal structure of healthy rabbits. Histopathological study failed to reveal the signs of toxic damage to the retina. The electroretinographic findings suggest that kenalog primarily affect the function of photoreceptors and Muller's glia. The fact that the alteration of retinal functional activity may be linked with the early cytotoxic effects of the agent cannot be ruled out. However, the changes found in retinal electrogenesis are most likely to reflect the mechanisms of kenalog's beneficial therapeutic action that is primarily mediated through its effect on the function of photoreceptors and Muller's cells. PMID- 17087034 TI - [The antiallergic eye drops "polynadyme": development, experimental and clinical studies]. AB - The antiallergic eye drops "Polynadyme", proposed by the Helmgolz Moscow Research Institute of Eye Diseases, have been prepared by the "Sintez" PJSC (Kurgan). The drops exert a combination of antihistaminic and vasoconstrictive effects and, for better tolerability, contain a low-toxic preserving complex. The drops are polymer-based, which ensures a long action and an artificial tear effect. Preclinical rabbit trials have shown the safety of the "Polynadyme" eye drops, their specific activity in preventing an allergic reaction, and their antiallergic effect on a model of allergic conjunctivitis. Comparative clinical trials covering 150 patients have yielded excellent and good results in 93% of cases. In acute allergic reactions, hyperemia, itch, and burning diminished just 5 minutes after administration. The "Polynadyme" eye drops are effective in treating pollinous conjunctivitis, spring (vernal) keratoconjunctivitis, allergic reactions when wearing contact lenses, the dry eye syndrome, drug-induced and toxicoallergic conjunctivitis, and other ocular allergic reactions. PMID- 17087035 TI - [The specific features of calculation of the focal power of intraocular lenses in microophthalmos]. PMID- 17087036 TI - [Serpiginous choroiditis (a clinical case)]. PMID- 17087038 TI - [A case of bilateral pupillary membrane]. PMID- 17087037 TI - [Non-concomitant strabismus. Proposals for its clinical classification]. PMID- 17087039 TI - [Congenital iridial colobomas]. PMID- 17087040 TI - [Negative remodeling of the retina in its degenerations]. PMID- 17087041 TI - [Non-ablative laser refractive interventions into the fibrous tunic of the eye]. PMID- 17087042 TI - The miracle of medicine--a gift to humanity. PMID- 17087043 TI - Suicide attempts by burning. AB - Setting oneself ablaze is an uncommon method of attempting suicide. Ten patients with deliberate thermal injuries had been evaluated by a consultation psychiatry service over a 15-year period. Schizophrenia and major depression were the more common diagnoses, and most patients had prior psychiatric histories. When compared to 1,864 people who attempted suicide by other means, the burn subjects were older, comprised a larger percentage of African Americans, and resembled those who had jumped from high places. Stresses encountered by recovering burn patients involved dependency, body image concerns, exacerbation of pre-existing depression, pain, flashbacks of the incident, and guilt intensified by their proximity to accidentally burned victims. Problems for burn unit staff included feelings of discomfort with difficult, potentially suicidal individuals, and responsibilities to perform painful procedures. Recommendations for psychiatric care include pharmacotherapy for depression or psychosis, visitation by previously self-burned individuals who serve as role models, patients' visualization of their injuries, and behavioral techniques to diminish reliance on analgesic drugs and to increase a personal sense of control. Suggestions for assisting staff include liaison support, personnel debriefing sessions, and inservice presentations on emotional aspects of burn management. PMID- 17087044 TI - Characteristics of participants enrolled in a Quit and Win smoking cessation program. AB - Quit and Win smoking cessation incentive programs have been used to promote community-based smoking cessation in the United States and internationally. The Quit and Win contest has been offered annually by the Lexington-Fayette County Health Department since 2001. In order to engage in continuous quality assessment of the Quit and Win program, demographic analysis of program participants is critical to better extend the marketing message to target audiences. The goal of this analysis is to report various baseline parameters of the 2003 Lexington Fayette County Quit and Win contest cohort. Of the 876 participants, most were white, had at least a high school education, smoked cigarettes for more than 10 years, and had multiple previous quit attempts. Most anticipated using nicotine replacement products to aid the current quit attempt. The demographic information from this analysis can be used to design future programs that target a wider segment of the population. PMID- 17087045 TI - What's with the bail-out? PMID- 17087046 TI - A campus conflict reveals rifts in deaf culture. PMID- 17087047 TI - Brush with perfection. PMID- 17087048 TI - Get the whole truth. PMID- 17087049 TI - Determinants of disuse-induced skeletal muscle atrophy: exercise and nutrition countermeasures to prevent protein loss. AB - Muscle atrophy results from a variety of conditions such as disease states, neuromuscular injuries, disuse, and aging. Absence of gravitational loading during spaceflight or long-term bed rest predisposes humans to undergo substantial loss of muscle mass and, consequently, become unfit and/or unhealthy. Disuse- or inactivity-induced skeletal muscle protein loss takes place by differential modulation of proteolytic and synthetic systems. Transcriptional, translational, and posttranslational events are involved in the regulation of protein synthesis and degradation in myofibers, and these regulatory events are known to be responsive to contractile activity. However, regardless of the numerous studies which have been performed, the intracellular signals that mediate skeletal muscle wasting due to muscular disuse are not completely comprehended. Understanding the triggers of atrophy and the mechanisms that regulate protein loss in unloaded muscles may lead to the development of effective countermeasures such as exercise and dietary intervention. The objective of the present review is to provide a window into the molecular processes that underlie skeletal muscle remodeling and to examine what we know about exercise and nutrition countermeasures designed to minimize muscle atrophy. PMID- 17087050 TI - Influence of oxidative stress on fusion of pre-synaptic plasma membranes of the rat brain with phosphatidyl choline liposomes, and protective effect of vitamin E. AB - Influence of oxidative stress on fusion of pre-synaptic plasma membranes with phosphatidylcholine (PC) liposomes as a model of synaptic vesicle was investigated. The inhibitory effect of vitamin E on the decline in the fusion caused by oxidative stress was also assessed. Rats subjected to hyperoxia as oxidative stress showed significant increases in the levels of lipid hydroperoxides and protein carbonyl moieties in pre-synaptic plasma membranes in the brain. The zeta potential of pre-synaptic membrane surface was decreased markedly. When synaptosomes were incubated with PC liposomes labeled by either rhodamine B or calcein as a fluorescence probe, or 12-doxyl stearic acid as an ESR spin trapping agent, translocation of each probe into oxidatively damaged pre synaptic membranes was decreased significantly. Fatty acid composition analysis in pre-synaptic membranes obtained from normal rats revealed a marked increase in linoleic acid and a moderate decrease in docosahexaenoic content after the incubation with liposomes. However, rats subjected to hyperoxia did not show marked changes in these fatty acid contents in their pre-synaptic membranes after the incubation. Such changes caused by hyperoxia were inhibited by vitamin E treatment of rats. These results suggest that oxidative damage of pre-synaptic membranes caused by oxidative stress lowers the lipid-mixing for the membrane fusion. The results of this study imply that vitamin E prevents the deficit in neurotransmission at nerve terminals due to the decline in fusion between pre synaptic membrane and synaptic vesicles caused by oxidative membrane damage. PMID- 17087051 TI - Effect of L-serine on the biosynthesis of aromatic amino acids in Escherichia coli. AB - It is well known that some amino acids inhibit bacterial growth. L-Serine is known to inhibit the growth of Escherichia coli by inhibition of homoserine dehydrogenase (EC 1.1.1.3). It has been reported that this L-serine inhibition may be prevented by the addition of L-isoleucine or L-threonine to the medium. In our study, however, recovery of the growth inhibition of Escherichia coli by L serine occurred in the presence of several amino acids, especially L phenylalanine. In an attempt to further elucidate this inhibition mechanism, different intermediates of aromatic amino acid biosynthesis were added to the growth medium. Recovery from the inhibition did not occur in the presence of prephenate but did occur when phenylpyruvate was added to the medium. The specific activity of prephenate dehydratase decreased in cells grown in the presence of L-serine. However. L-serine did not inhibit in vitro prephenate dehydratase activity, and the expression of pheA, which encodes the prephenate dehydratase, was not depressed by L-serine. We suggest that L-serine acts via another inhibition mechanism. Although this inhibition mechanism has not been fully elucidated, our results suggest that the addition of L-serine to the growth medium inhibits prephenate dehydratase synthesis and thus affects L-phenylalanine biosynthesis. PMID- 17087052 TI - The effects of eicosapentaenoic acid-fortified food on inflammatory markers in healthy subjects--A randomized, placebo-controlled, double-blind study. AB - Epidemiological studies showed that habitual fish intakes were associated with lower blood inflammatory markers. In the present study the effects of a fish oil containing food on inflammatory markers were investigated in healthy, mostly middle-aged subjects (59 men and 82 women) with normal to mildly elevated triglyceride levels. Study subjects were randomly allocated to two groups in a double-blind manner; one group ingested an eicosapentaenoic acid (EPA)-rich fish oil-fortified drink (0.60 g EPA+0.26 g docosahexaenoic acid/d. EPA group, n=68) for 12 wk. The rest of the subjects took a placebo (control group, n=73). Plasma levels of high sensitivity C-reactive protein (hs-CRP) and soluble tumor necrosis factor-receptors 1 and 2 (sTNF-Rs 1 and 2) were measured at the start and end of intervention. EPA concentrations in the total RBC phospholipid fraction significantly increased by 79% in the EPA group at the end of the study, and they changed very little in the control group (+0.68%). The inflammatory markers did not change in either group. It is likely that fish oil does not change hs-CRP or sTNF-Rs 1 or 2 in subjects without active inflammation. PMID- 17087053 TI - Favorable effects of tea on reducing the cognitive deficits and brain morphological changes in senescence-accelerated mice. AB - The present study was carried out to explore the effects of oolong and green teas on improving the memory deficits and brain pathological changes in senescence accelerated-prone mice P8 (SAMP8). Six-month-old mice were supplied with oolong tea, green tea or water as the sole drinking fluid for 16 wk. The memory ability of mice was evaluated by passive and active avoidance tests, while the extent of the brain degeneration was measured by the spongiosis grades and the lipofuscin percentage in the hippocampus. The total grading score and serum biochemical levels were also measured. The results indicated that the mice supplemented with the oolong and green tea drinks reversed the cognitive impairment, lessened the spongy degeneration and lipofuscin, and increased the serum Trolox equivalent antioxidant capacity more than the control group. The total grading score of the oolong tea group was lower than that of the control group in male mice, whereas it did not differ among female groups. No differentiations in the concentrations of total cholesterol. triglyceride, glucose, iron or hemoglobin were observed among three drink groups. In conclusion, oolong and green teas could reduce the deteriorations of cognitive ability, brain degenerative changes and aging process in SAMP8, probably through the potent antioxidative activity of the tea. PMID- 17087054 TI - The protective effect of the soybean polyphenol genistein against stress-induced gastric mucosal lesions in rats, and its hormonal mechanisms. AB - The present study investigated the effect of the soybean polyphenol genistein on the stomach using a water immersion restraint (WIR) stress model. Male Wistar rats were administered 50 or 100 mg/kg/d of genistein for 2 wk or were not given any drug. Rats were subjected to WIR stress for 4 h. At the end of the WIR period, rats were sacrificed. Subsequently, rats underwent measurement of the ratio of the mucosal hemorrhagic erosion area to the whole stomach body area, myeloperoxidase (MPO) activity, superoxide dismutase (SOD) activity, thiobarbituric acid reactive substances (TBARS) level, and proinflammatory cytokines (tumor necrosis factor (TNF)-a and cytokine-induced neutrophil chemoattractant (CINC)-1) levels in the gastric tissue. Furthermore, an isolated rat stomach infusion model was employed for the endocrinological investigation of the effect of genistein. The extracted stomach canal and the vascular system, which comprised the experimental model, were subjected to perfusion. After 20 min of perfusion, the perfusate from the portal vein was collected, and the concentrations of histamine, gastrin, and somatostatin in the perfusate were measured. Experiments demonstrated that genistein administration resulted in significant suppression of WIR stress-induced gastric mucosal injury and MPO activity, Further, genistein significantly elevated SOD activity and significantly suppressed the TBARS level, production of TNF-alpha and CINC-1, and secretion of gastrin, histamine, and somatostatin. These data suggest that genistein protected against gastric mucosal injury, likely via its ability to inhibit oxidation, inflammation, and secretion of gastrin and histamine. PMID- 17087055 TI - Comparison of body fat distribution and blood lipid profiles according to Trp64Arg polymorphism for the beta 3-adrenergic receptor gene in Korean middle aged women. AB - The aim of this study was to determine whether there was an association between body fat distribution, blood lipid profiles, and beta 3-adrenergic receptor gene polymorphism in Korean middle-aged women. Subjects were grouped according to BMI as obese (> or = 5 BMI, n = 95) or non-obese (BMI<25, n = 93). The Trp64Arg mutation of the beta 3-adrenergic receptor gene was detected by PCR-RFLP. Skinfold thickness, body circumference, intra-abdominal fat area by CT, and blood lipid profiles were also measured. Data were compared using ANOVA, Bonferroni t test, and Chi-square. Significance for statistical analyses were set at p<0.05. In the obese group, 63.16% were Trp64Trp homozygotes and 36.84% were Trp64Arg heterozygotes, compared to 80.65% who were Trp64Trp homozygotes and 19.35% who were Trp64Arg heterozygotes in the non-obese group. These results indicated a significant (chi-square = 4.943, p<0.05) difference between the two groups. Frequency of the Arg64 allele in the obese group (16.84%) showed a significant (chi-square = 4.185, p<0.05) difference as compared to the non-obese group (9.68%). Skinfold thickness and body circumference of the Trp64Arg heterozygote group showed a consistent increase as compared to the Trp64Trp homozygote group. Visceral fat area and VSR of Trp64Arg heterozygote group showed a higher tendency than Trp64Trp homozygotes in the obese group, but these differences were not statistically significant. In conclusion, the Trp64Arg polymorphism of the beta 3 adrenergic receptor gene is associated with obesity in middle-aged Korean women, but it is difficult to suggest the prominent association of the Trp64Arg polymorphism of the beta 3-adrenergic receptor gene with prevalence of abdominal obesity or dyslipidemia in Korean middle-aged women. PMID- 17087056 TI - Effects of fenugreek seeds (Trigonella foenum greaecum) extract on endurance capacity in mice. AB - The present study was designed to determine the effect of fenugreek seed extract (FG) on endurance capacity in male mice aged 4 wk. Mice were given orally either vehicle or FG (150, 300 mg/kg body weight) by stomach intubation for 4 wk. The 300 mg/ kg FG group showed a significant increase in swimming time to exhaustion as compared to the control group. In the FG groups, blood lactate concentration was significantly lower than in the control group. In the control group, plasma non-esterified fatty acid (NEFA) and plasma glucose were decreased by swimming exercise. But in the FG group, NEFA and plasma glucose were significantly increased by swimming. FG treatment also significantly decreased fat accumulation. These results suggest that improvement in swimming endurance by the administration of FG is caused by the increase in utilization of fatty acids as an energy source. PMID- 17087057 TI - Genistein suppresses development of spontaneous atopic-like dermatitis in NC/Nga mice. AB - In this study, we examined the effect of genistein on the severity of dermatitis and level of serum IgE in NC/Nga (NC) mice. NC mice housed in conventional conditions develop spontaneous atopic-like dermatitis; however, oral administration of 20 mg/kg genistein suppresses the development of dermatitis. We also investigated the levels of serum IgE in genistein-treated NC mice and found that the levels were the same as those in control NC mice. We further investigated in vitro IFN-gamma and IL-4 production from spleen cells upon stimulation with anti-CD3 and anti-CD28 mAbs. IFN-gamma production level in NC mice that received 20 mg/kg genistein was significantly lower than that in control NC mice. In contrast, the production level of IL-4 in genistein-treated mice was not significantly different from that in control mice but tended to increase in a dose-dependent manner. PMID- 17087058 TI - [Antiviral etiotropic chemicals: efficacy against influenza A viruses A subtype H5N1]. AB - The paper analyzes data of an experimental study of the efficacy of antiviral agents (amantadine, remantadine, ozeltamivir, zanamivir, arbidol, ribavirin) in the cultured cells and on a model of murine influenza pneumonia against influenza A viruses subtype H5N1. It also gives data on their use in the treatment of human beings during avian influenza outbreak. The mechanism of action of the agents, pharmacokinetics, adverse reactions, and their potential resistance are considered. PMID- 17087059 TI - [Hemorrhagic (Marburg, Ebola, Lassa, and Bolivian) fevers: epidemiology, clinical pictures, and treatment]. AB - The evaluation of the biological and epidemiological properties of Ebola, Marburg, Lassa, and Machupo viruses suggests that they are of social importance for health care authorities. The studies have created prerequisites to the development of reliable biosafety means against these pathogens. Particular emphasis is laid on the methods for infection diagnosis and on the studies to design specific protective agents--immunoglobulins and inactivated vaccines. PMID- 17087060 TI - [Molecular mechanisms of reversions to the ts+ -phenotype of cold-adapted influenza virus A strains--attenuation donors for live influenza reassortant vaccines]. AB - A ts+ revertant of cold-adapted (ca) strain A/Leningrad/134/47/57--the attenuation donor for live influenza reassortant vaccines--was obtained by passages of the ca strain in chick embryos at nonpermissive temperatures. The ts+ revertant acquired the ability to grow in chick embryos at 40 degrees C and lost the capacity to reproduce there at 25 degrees C. A complementation-recombination test using the fowl plague virus (FPV0 ts-mutants showed the loss of the ts phenotype in the RNA-segments of ts+ revertants' genome coding for PB2, NP, and NS (NS2) proteins. However, PCR-restriction analysis revealed a true reversion in RNA-segment coding for PB2 protein only. All the investigated mutations in the ts+ revertant genome were preserved. This phenomenon could be explained by the appearance of intragenic and extragenic suppression mutations in the ts+ revertant genome. The data of the complementation-recombination test suggest that reversion of ts-phenotype occurs more frequently due to extra- or intragenic suppression rather than as a result of a true mutation loss. Estimation of the genetic stability of vaccine ca strains of influenza virus should be based on the combined use of PCR-restriction and complementation tests. PMID- 17087061 TI - [Immune response in vaccinated and non-vaccinated mice infected with tick-borne encephalitis virus]. AB - The paper deals with an investigation of an immune response in BALB/c mice immunized with tick-borne encephalitis (TBE) vaccine and infected with TBE virus and in non-immunized mice. The parameters of specific humoral (IgG and IgM) and cellular (gamma-interferon (IFN) and cell proliferation) immunities and the activity of cytokines (necrosis tumor factor-alpha, interleukin (IL)-1beta, IL-2, IL-6, IL-10, and IL-12) were studied. There were significant differences in the specific and nonspecific immune response of immunized and non-immunized animals. Noteworthy is the difference in the time course of changes in the levels of IL-6, IL-2, IL-12, and gamma-IFN in the immunized and non-immunized animals. PMID- 17087062 TI - [Epizootology of hemorrhagic fever with renal syndrome in the Central Chernozem Region]. AB - A total of 5149 small mammals belonging to 16 species were collected from the Lipetsk, Voronezh, and Belgorod regions (40 administrative districts) in 2003 2004 and examined by ELISA and IFA to detect hantavirus antigen and antibodies in the lung tissues. Hantavirus circulation was revealed in 13 species, the highest hantavirus activity being detected in field (Apodemus agrarius) and small wood (A. (S) uralensis) mice (Dobrava-Belgrad virus), bank (Clethrionomis glareolus) (Puumala virus) and common (Microtus arvalis) (Tula virus) voles. These species were frequently found to have their untypical hantaviruses, which was most commonly observed in small wood mice. It is suggested that the small wood mouse is likely to take a certain part in maintaining the circulation of Dobrava Belgrad virus. PMID- 17087063 TI - [Molecular virological monitoring of Newcastle disease virus strains (Paramyxoviridae, Avulavirus) in the populations of wild birds in the Volga estuary (the 2001 data)]. AB - The paper analyzes the results of isolation of Newcastle disease virus (NDV) strains from 336 swaps of 31 wild bird species collected in the 2001 summer in the Volga estuary (Astrakhan Region). Twenty-seven NDV strains were isolated from little terns (Sterna albifrons) (n=11; infection rate, 24.4%), great cormorants (Phalacrocorax carbo) (n=6; 11.1%), coots (Fulica atra) (n=8; 6.5%), sandwich terns (Sterna sandvicensis) (n=1; 100%), and common redshanks (Tringa totanus) (n=1; 50.0%). Four strains were sequenced by the 374 n. a. residue fragment from the beginning of the F gene, one of them was by the full F gene, and another (Stemal/Astrakhan/2755/2001) was by the full genome. Nucleotide sequences have allowed the authors to classify corresponding NDV strains as 5b genotype and the analysis of the amino acid sequence of the F-protein cleavage site has shown them to belong to a non-pathogenic group. PMID- 17087064 TI - [Phenotyte of epidemic influenza B virus strains isolated in different years]. AB - The reproducing ability at elevated temperatures (non-ts phenotype) was examined for 38 influenza B virus strains isolated in different years in different countries. Out of the 7 strains isolated in 1940 to 1973, only one showed temperature-sensitivity of reproduction (a ts phenotype). In 1984 to 1988, the proportion of temperature-sensitive strains increased up to 55% (6 of 11). Since the late 1990s, the majority (90%) of the study influenza B viruses demonstrated a pronounced ts phenotype. Influenza B virus strains were also examined for their resistance to serum inhibitors. Prior to the divergence of influenza B viruses into two lines: B/Jamagata and B/Victoria, the epidemic viruses exhibited a high resistance to nonspecific inhibitors of normal equine serum. This property was also preserved in all study B/Victoria strains; however, 83% of the B/Jamagata viruses were inhibitor-sensitive. The present study has demonstrated the heterogenicity of epidemic influenza B viruses in temperature- and inhibitor sensitivity. PMID- 17087065 TI - [Antiviral activity of Bacillus intermedius RNAase in guinea-pigs and rabbits infected with outdoor rabies virus]. AB - Single injection of Bacillus intermedius RNAase in a dose of 5 mg/kg could protect 40 and 50-70% of the outdoor rabies virus-preinfected guinea-pigs and rabbits, respectively. In the control group there were 100 and 75-100% deaths of the RNAase-untreated guinea-pigs and rabbits, respectively. Animal protection was observed only when RNAase was injected into the site of viral administration. The intramuscular injection of RNAase, other than the site of viral administration failed to protect the infected animals. The efficacy (75%) of RNAase injected into the rabbits was similar 1 and 24 hours after animal infection. PMID- 17087066 TI - [Enzyme immunoassay for detection of porcine circovirus type 2, by using the recombinant capsid protein ORF-2]. AB - Recombinant antigen ORF2 from porcine circovirus type 2 (PCV-2) was produced, by using the baculovirus expression system, with histidine tags to allow purification by metal-chelate affinity chromatography. The purity of the protein was verified by polyacrylamide gel electrophoresis; and its immunospecificity was confirmed by the immunoblotting test using reference PCV-2-positive and PCV-2 negative porcine sera and monoclonal antibodies. The protein was used as an antigen to develop an indirect enzyme immunoassay (EIA) of PCV-2 antibodies. EIA was shown to have a high sensitivity and specificity as compared with indirect immunofluorescence test. Porcine serum samples from 15 pig-breeding farms of the Russian Federation were studied. Seropositive samples were found in all age pig groups in all the farms, The number of seropositive animals was shown to be directly related to its age. PMID- 17087067 TI - [Poliomyelitis virus cannot be eradicated. Why?]. PMID- 17087069 TI - The second century--political involvement. PMID- 17087068 TI - Taking a hint... PMID- 17087070 TI - Antimicrobial symposium. Part 1. Principles of antimicrobial therapy. PMID- 17087071 TI - Milk-alkali syndrome in pre-eclamptic pregnancy: report of a patient and evaluation of albumin-corrected calcium in pre-eclamptic pregnancies. AB - BACKGROUND: First reported in 1923 due to excessive ingestion of milk and bicarbonate for peptic ulcer disease, milk-alkali syndrome nearly disappeared by the 1980s. More recently, however, this syndrome has become a more common cause of hypercalcemia. This increase is likely due to the increased use of calcium carbonate for the prevention and treatment of osteoporosis. Pregnancy likely places women at risk for milk-alkali syndrome due to increased intestinal absorption of calcium. Recommendations for increased oral calcium intake during pregnancy along with frequent use of calcium carbonate for GI symptoms during late pregnancy increase this risk. METHODS: We performed a retrospective chart review of pregnant women with pre-eclampsia, ages 18-35 years, who were admitted for delivery to the University of Oklahoma Health Sciences Center. Serum calcium levels were reviewed and compared with those of an age- and sex-matched control group consisting of 100 non-pregnant women admitted to the same facility. RESULTS: Twenty-nine of the 100 pregnant patients (29%) had an albumin-corrected calcium above normal compared to only six patients (6%) in the control group (P = 18.32, p < 0.0001). The highest corrected serum calcium in the pregnant group was 12.3 mg/dl compared to 10.7 mg/dl in the control group. SUMMARY: Milk-alkali syndrome is not an uncommon cause of hypercalcemia, and pregnancy may predispose women to milk-alkali syndrome as demonstrated by the patient reported. Albumin corrected calcium is frequently high in women with pre-eclampsia but ionized calcium is not. PMID- 17087072 TI - Medicaid grievance resolution. Past, present, and future. PMID- 17087073 TI - Insuring compliance with the physician "immediately available" requirement. PMID- 17087074 TI - [Modern biological basis of Chinese medical theory that "kidney nourishes marrow and brain is sea of marrow"]. AB - The theory that "the kidney nourishes marrow and brain is the sea of marrow" has been instructing traditional Chinese medical doctors in preventing and treating dementia in aged people for thousands of years. However, the modern biological basis of this theory has not been systemically studied. In this review, we summarized our serial pharmacological studies on the treatment of Alzheimer's disease (AD) with Chinese medicinal herbs, using multiple kinds of AD-like animal and cell models. The results show that "kidney-reinforcing" herbs of traditional Chinese medicine play a very important role in the anti-AD effects; and different "kidney-reinforcing" herbs have different effects in brain functions. The common effects of "kidney-reinforcing" herbs are improving cellular energy metabolism, increasing neurotrophic factors and the number of cholinergic neurons, and decreasing neurotoxin production. Based on above results, we propose that the essence of "brain marrow" is the neurons and neurotrophic factors in the brain; "the deficiency of brain marrow" is induced by the decrease in neurotrophic factors and the atrophy and loss of neurons in the brain, thus resulting in cognitive impairment and dementia. The modern biological basis of "reinforcing kidney to replenish marrow" by traditional Chinese medicine includes improving cellular energy metabolism and utilization, enhancing endogenous neurotrophic effects and decreasing neurotoxin production, thus reducing the cell death and increasing the survival and regeneration of neurons. PMID- 17087075 TI - [Comparison on growth, physiology and medicinal components of Dendrobium huoshanense hybrid and its parents]. AB - OBJECTIVE: To compare the hybrid between species of Dendrobium huoshanense and its parents on growing, physiologic indexes and content of medicinal components, and provide theoretical basis for species quality improvement. METHOD: The chlorophyll content, the photosynthesis rate, the polysaccharides content and the alkaloids content were measured by anhydrous ethanol method, Cl-310 photosynthesis determination system, colorimetry of concentrated sulphuric acid phenol and acid dyes colorimetry respectively. RESULT: The growth of hybrid was close to D. moniliforme, and apparently higher than D. huoshanense. The chlorophyll content and the photosynthesis rate of one-year-hybrid were markedly higher than its parents. The content of polysaccharides and alkaloids in two-year stem and three-year-stem of hybrid were close to that of D. huoshanense. CONCLUSION: The hybrid integrates superiority of parents on growth and accumulation of medicinal components opens vast vistas for development and utilization. PMID- 17087076 TI - [Studies on cytogeography of Pinellia ternata poliploid complex]. AB - OBJECTIVE: In order to clarify the genetic background of Pinellia ternata germplasm resources in China, the chromosomal constitution and cytogeographical distribution of P. ternata were investigated in 27 different populations among 16 provinces and regions in China systematically. METHOD: Cytological and cytogeographical methods were used in the study. RESULT: P. ternata in China is a polyploid complex, which contains septuploid (2n = 7x = 91) , octoploid (2n = 8x = 104) , nonuploid (2n = 9x = 117) and decaploid (2n = 10x = 130). Meanwhile the aneuploid series (2n = 92, 103, 105, 115) of a minority of P. ternata were also found. CONCLUSION: The genetic differentiation and the phenomenon of ploidy miscellany commonly exist in the species of P. ternata in China, both for natural populations and cultivated populations. Toxicity and chemical components of different ploidy P. ternata should be clarified before the superior multiploid is selected for normalized plantation of the plant. PMID- 17087077 TI - [Studies on in vitro culture of adventitious root in Salvia miltiorrhiza]. AB - OBJECTIVE: To study the culture of adventitious root of Salvia miltiorrhiza in vitro systemically. METHOD: Effects of sucrose concentrations, medium pH, inoculum size and plant growth regulators on adventitious root growth and secondary metabolites production in S. miltiorrhiza were investigated. RESULT: With the increase of initial sucrose concentration, adventitious root growth rates increased and tanshinone II A content decreased, while content of protocatechuic aldehyde showed a broken line change and its highest production was obtained under 30 g x L(-1) sucrose in the medium. As for the effect of medium pH, medium pH of 6.5, 5.5 (or 6.0) and 5.8 was favorable for adventitious root growth, tanshinone II A and protocatechuic aldehyde synthesis respectively. Furthermore, adventitious root growth, rate was greatly increased when inoculum size was 2.5%. MS medium added with 0.5 mg x L(-1) KT was much favorable for tanshinone II A and protocatechuic aldehyde accumulation. CONCLUSION: Parameters including sucrose concentrations, medium pH, inoculum size and plant growth regulators have distinct effects on the in vitro culture of adventitious root growth and secondary metabolites synthesis of S. miltiorrhiza. PMID- 17087078 TI - [Studies on preparation of sustained-release Shuxiong formulation, a traditional Chinese medicine compound recipe, using time-controlled release techniques]. AB - OBJECTIVE: To prepare a sustained-release formulation of traditional Chinese medicine compound recipe by adopting time-controlled release techniques. METHOD: Shuxiong tablets were chosen as model drug. The prescription and technique of core tablets were formulated with selecting disintegrating time and swelling volume of core tablets in water as index. The time-controlled release tablets were prepared by adopting press-coated techniques, using PEG6000, HCO and EVA as coating materials. The influences of compositions, preparation process and dissolution conditions in vitro on the lag time (T(lag)) of drug release were investigated. RESULT: The composition of core tablets was as follow: 30% of drug, 50% MCC and 20% CMS-Na. The T(lag) of time-controlled release tablets was altered remarkably by PEG6000 content of the outer layer, the amount of outer layer and hardness of tablet. The viscosity of dissolution media and basket rotation had less influence on the T(lag) but more on rate of drug release. CONCLUSION: The core tablets pressed with the optimized composition had preferable swelling and disintegrating properties. The shuxiong sustained-release formulations which contained core tablet and two kinds of time-controlled release tablets with 3 h and 6 h of T(lag) could release drug successively at 0 h, 3 h and 6 h in vitro. The technique made it possible that various components with extremely different physicochemical properties in these preparations could release synchronously. PMID- 17087079 TI - [Study on influence of processing methods on chemical constituents in Radix Paeoniae Alba]. AB - OBJECTIVE: The influence of processing methods on chemical constituents in Radix Paeoniae Alba was observed. METHOD: A HPLC method was used for analyzing the changes of eight major constituents, namely gallic acid, paeoniflorin sulfonate, catechin, paeoniflorin sulfonate, albiflorin, paeoniflorin, benzoic acid, pentagalloylglucose and benzoylpaeoniflorin, with the three processing procedures of decorticating, boiling and fumigating by burning of sulphur. Analysis was performed using a Zorbax SB-C18 column (4.6 mm x 250 mm, 5 microm) with the mixture of acetonitrile (A) and 0.015% phosphoric acid solution as mobile phase in gradient mode. The detection wavelength was set at 230 nm and the column temperature was at 30 degrees C. RESULT: Except for gallic acid and pentagalloylglucose, the other constituents decreased during procedure of decorticating and boiling. Fumigating by burning of sulphur would produce a new compound, paeoniflorin sulfonate, which was a byproduct from the reaction of paeoniflorin with SO2. CONCLUSION: The significant changes were produced in chemical constituents of Radix Paeoniae Alba during three processing procedures. Therefore, the processing of Radix Paeoniae Alba should be strictly controlled and standardized. PMID- 17087080 TI - [Application of HPLC fingerprint in quality control of Fufangteng combination formula]. AB - OBJECTIVE: To establish the characteristic mode of HPLC fingerprint of Fufangteng combination formula for the quality control of this medicine. METHOD: 10 batch certified products were selected to establish the characteristic mode of HPLC fingerprint of Fufangteng combination formula by chromatographic pattern matching and retention time correction. RESULT: This characteristic mode of HPLC fingerprint can clearly differentiate the eligible Fufangteng products from the unqualified products. CONCLUSION: This method can provide thorough information for the quality control of Fufangteng combination formula. PMID- 17087081 TI - [Determination of three chemical components in Fructus aurantii immaturus]. AB - OBJECTIVE: To determine the contents of 3 kinds of components in Fructus aurantii immaturus. METHOD: HPLC analysis was performed to detect the contents of hesperidin, naringin and synephrine. The content of volatile oil was detected determined following the method of Chinese pharmacopoeia. RESULT: The contents of hesperidin, naringin, synephrine and volatile oil in ten samples are from 1.25% to 16.6%, 0% to 13.9%, 0.058 5% to 0.676% and 0.1% to 2.2%, respectively. CONCLUSION: The contentre are significant differences of among chemical components in from different samples of Fructus aurantii immaturus are greats. PMID- 17087082 TI - [Determination of total flavonoids in Abrus cantoniensis and its dynamic changes]. AB - OBJECTIVE: To develop a method for ultrasonic extraction and determination of total flavonoids in Abrus cantoniensis, and to analyze its dynamic changes. METHOD: The optimized condition of extraction of total flavonoids was studied with orthogonal design. The contents of total flavonoids in different organs and of different growth stages were determined by UV-visible spectrophotometer. RESULT: The ethanol volume and extraction times were the main factors impacting the effects of ultrasonic extraction. The content of total flavonoids in stems were higher than in roots and the lowest in leaves. The dynamic changes of total flavonoids contents in roots and stems of A. cantoniensis were in similar trends. Its total flavonoids content in the two parts of plant increased gradually with the growth and reached the maximum in October, and the content decreased significantly in Feburay of next year. The content of total flavonoids in leaves reached also to the highest value before leaves fell off. CONCLUSION: The optimized extraction method of total flavonoids in Abrus contoniensis was obtained with three times with 80% ethanol at 20 times of volume for 30 min. The results implied that the best yield and quality may be obtained before leaves fall. PMID- 17087083 TI - [Study of quality standards on Qianshan Huoxue plaster]. AB - OBJECTIVE: To establish the methods of identification and assay in Qianshan Huoxue Gao. METHOD: Using TLC to identify Sanchi, Dragon's Blood and using HPLC to determine the content of ginsenoside Rg1. RESULT: The linear range of ginsenoside Rg1 was from 0.153 9 to 1.026 microg. The average recovery was 97.4%, RSD was 2.1%. CONCLUSION: The methods are simple and have good reproducibility. PMID- 17087084 TI - [Studies on triterpenoids from Potentilla chinensis]. AB - OBJECTIVE: To investigate the chemical constituents of Potentilla chinensis. METHOD: Silica gel column chromatography and Sephadex LH - 20 gel column chromatography were employed for the isolation and purfication. The structures were identified on the basis of spectral data and chemical evidence. RESULT: Six compounds were isolated and identified as follows: 3-hydroxy-11-ursen-28, 13 olide, 11, 12-dehydroursolic acid lactone (1), 3-O-acetyl pomolic acid (2), betulinic acid (3), 3-oxo-12-ursen-28-oic acid (4), ursolic acid (5), oleanic acid (6). CONCLOUSION: All these compounds were isolated from P. chinensis for the first time, compound 1, 2, 4 were isolated from this genus for the first time. PMID- 17087085 TI - [Studies on chemical constituents of Paederia scandense]. AB - OBJECTIVE: To study the chemical constituents of Paederia scandense. METHOD: The constituents were isolated and purified by silica gel and Sephadex LH - 20 column chromatography. Their structures were elucidated by physicochemical properties and spectral analysis. RESULT: 20 compounds were obtained and identified as rubiadin-1-methylether (1), diadzein (2), cleomiscosin B (3), cleomiscosin D (4), isolariciresinol (5), linarin (6), isoscopoletin (7), caffic acid (8), coumarinic acid (9), p-hydroxyl-benzoic acid (10), oleanolic acid (11), ursolic acid (12), beta-sitosterol (13), daucosterol (14), paederoside (15), paederosidic acid (16), paederosidic acid methyl ester (17), saprosmoside E (18), paederoscandoside (19), caffeic acid 4-O-beta-D-glucopyranoside (20). CONCLUSION: Compounds 1-10, and 20 were isolated from this plant for the first time. PMID- 17087086 TI - [Chemical constituents of unsaponifiable matter from seed oil of Momordica cochinchinensis]. AB - OBJECTIVE: To study the chemical constituents of unsaponifiable matter from the seed oil of Momordica cochinchinensis. METHOD: The fatty oil from the seeds of M. cochinchinensis was extracted with petroleum ether, and the saponification was carried out with potassium hydroxide. The unsaponifiable matter was isolated and purified by silica gel column chromatography, and the structures of their constituents were elucidated by means of IR, MS, 1H-NMR, and authentic chemicals. RESULT: Karounidiol (1), isokarounidiol (2), 5-dehydrokarounidiol (3), 7 oxodihydrokarounidiol (4), beta-sitosterol (5), stigmast-7-en-3beta-ol (6), and stigmast-7,22-dien-3beta-ol (7) were elucidated. CONCLUSION: These compounds were found in this plant for the first time. PMID- 17087087 TI - [Study on ingredients of essential oils of Curcuma wenyujin extracted by supercritical-CO2 fluid extraction and steam distillation]. AB - OBJECTIVE: To compare the ingredients of essential oils of Curcuma wenyujin extracted by supercritical-CO2 fluid extraction and by steam distillation. METHOD: GC-MS was applied in this experiment. RESULT: The ingredients and physical and chemical properties of essential oils of C. wenyujin extracted by supercritical-CO2 fluid extraction and by steam distillation are similar. CONCLUSION: Supercritical-CO2 fluid extraction is better than steam distillation in extraction time, power consumption, recovery and purity. PMID- 17087088 TI - [Effects of xuezhikang and simvastatin on cerebral ischemia-reperfusion injury in rat]. AB - OBJECTIVE: To observe the effects of Xuezhikang and simvastatin on cerebral ischemia/reperfusion injury in rat, as well as the influences after intervention with L-NAME. METHOD: Rats were given orally with Xuezhikang and simvastatin or vehicle for 2 weeks, and then subjected to middle cerebral artery occlusion for 120 min using intraluminal filament model. L-NAME were injected into the lateral ventricles in half of the rats treated with Xuezhikang and simvastatin 45 min before the ischemia. The neurological deficits examinations were performed at 2, 24, 48 h after reperfusion. After the last examination the animals were sacrificed, the infarct volumes were determined by TTC staining, and MDA levels were also measured. RESULT: Xuezhikang and simvastatin both significantly reduced the infarct volume and improved the functional recovery when compared to vehicle. Xuezhikang and simvastatin both significantly decreased the MDA accumulation after reperfusion. L-NAME partially inhibited the protective effect of Xuezhikang but nearly completely abolished the protective effect of simvastatin. CONCLUSION: Xuezhikang has protective effects on ischemic brain damage in rats, which the beneficial effects are partly due to the statins components. The other components in Xuezhikang may also account for the neuroprotective effects, which is worth further investigations. PMID- 17087089 TI - [Antitumor effects of novel triterpene from Celastrus hypoleucus on human colorectal cancer cell line RKO in vitro]. AB - OBJECTIVE: To investigate the effects of novel triterpene (12-oleanene-3beta, 6alpha-diol) from Celastrus hypoleucus on the proliferation and apoptosis of human colorectal cancer cell line RKO. METHOD: The inhibitory effect of the novel triterpene on RKO cell proliferation was assayed by MTT dye reduction. The morphology of apoptotic cells was observed with AO/EB double fluorescence staining and HE staining, DNA fragment with electrophoresis on agarose gels, sub diploid peak and cell cycle with flow cytometer (FCM). RESULT: Novel triterpene (12-oleanene-3beta, 6alpha-diol) from C. hypoleucus significantly inhibited proliferation of RKO cells in dose-dependent and time-dependent manner, the IC50 was (12.20 +/- 0.79) microg x mL(-1) at 48 h. Typical apoptotic changes were observed in RKO cells under the fluorescence microscope and the light microscope. DNA ladder was detected on agarose gels at concentrations from 10 microg x mL(-1) to 20 microg x mL(-1) at 48 h. With FCM methods, dose-dependent apoptosis-induced effect was observed in RKO cell line after treatment of triterpene for 48 h, and the apoptotic rates were increased from(2.93 +/- 0.84) % to (50.79 +/- 6.61) % at concentrations from 2.5 microg x mL(-1) to 20 microg x mL(-1). DNA histograms data from FCM analysis showed that the number of cells was obviously reduced during G0-G1 phase and G2-M phase, but not during S phase for RKO cell line after treatment with various concentrations of the triterpene for 48 hours. CONCLUSION: Novel triterpene (12-oleanene-3beta, 6alpha-diol) from C. hypoleucus can induce apoptosis and has inhibition effect on the proliferation in RKO cell line. PMID- 17087090 TI - [Effects of jianwei yuyang granule on inflammatory reaction and NF-kappaB expression in rat gastric mucosa of ulcer healing and recurrence]. AB - OBJECTIVE: To observe the effects of Jianwei Yuyang granule (JWYY) on inflammatory reaction and NF-kappaB expression in rat gastric mucosa of ulcer healing and recurrence. METHOD: Gastric ulcer was induced in rat by acetic acid according Okeba's method with some modification and the recurrence model was induced by IL-1beta. Pathohistology of ulcer healing and recurrence was observed. Density of inflammatory cell infiltrating regenerative mucosa, NF-kappaB protein and mRNA expression were measured. RESULT: JWYY had effects on improving the quality of ulcer healing, reducing the rate of ulcer recurrence, decreasing the density of inflammatory cell infiltrating regenerative mucosa and suppressing the activation and expression quantity of NF-kappaB protein and mRNA. CONCLUSION: JWYY may promote the ulcer healing and prevent the recurrence of the gastric ulcer by suppressing the activation of NF-kappaB and the following inflammatory reaction. PMID- 17087091 TI - [Inhibitory effect of weikangfu recipe on growth of mouse S180 tumor and its apoptotic induction]. AB - OBJECTIVE: To study the growth inhibition of Weikangfu recipe on S180 tumor and its apoptotic induction. METHOD: S180-bearing mice were orally administrated with different dosages of Weikangfu recipe, and the growth inhibition was evaluated; apoptotic cells induced were detected by flow cytometry and DNA agarose gel electrophoresis. RESULT: Weikangfu recipe showed significant inhibition on the growth of S180 tumor in a dose-dependent manner, compared with the control group. From apoptotic analyses, Weikangfu recipe induced a dose-dependent apoptosis of S180 tumor cells and arrested the cell cycle distribution at G0-G1 phase. At the same time, the up-regulation of p53 and bax and down-regulation of bcl-2 were observed in S180 tumor cells of the treated groups. CONCLUSION: Our findings demonstrate that Weikangfu recipe can significantly inhibit the growth of S180 tumor and induce apoptosis through expression alteration of p53, bax and bcl-2. PMID- 17087092 TI - [Study on antitumor activities of huanglian jiedu decoction]. AB - OBJECTIVE: To study the antitumor activity of Huanglian Jiedu decoction (HLJDT). METHOD: Antitumor activities were tested in mice with experimental tumor H22 in vivo, and the thymus index, spleen index and tumor inhibitory rate were evaluated. The effects on cancer cells from human were investigated in vitro using serum pharmacological approach. Swille, SPC-A-1, SGC-7901 and MCF-7 cancer cells were incubated in culture media containing serum from mice medicated with HLJDT. The inhibitory effects of HLJDT serum were observed by MTT assay. RESULT: HLJDT showed significant antitumor activities on H22 in mice. All of the HLJDT serum in different dosage groups could highly inhibit the proliferation of 4 cancer cell lines from human. CONCLUSION: The HLJDT can significantly inhibit the tumor H22 in mice in a dose-dependent manner, the drug serum has obvious anticancer effects against Swille, SPC-A-1, SGC-7901 and MCF-7. PMID- 17087093 TI - [Mechanism of wenpi decoction delaying progression of chronic renal failure]. AB - Wenpi decoction ameliorates the harmful factors of chronic renal failure, such as mesangial matrix and its cytokines, nuclear factor, endothelium-dependent relaxing factor, oxidative free raidical, apoptosis, lipide, protein, and blood rheology. Wenpi decoction is an effective agent for delaying the progression of chronic renal failure. PMID- 17087094 TI - Methicillin resistant Staphylococcus aureus. PMID- 17087095 TI - The immunomodulatory effect of antibiotics on the secretion of tumour necrosis factor alpha by peripheral blood mononuclear cells in response to Stenotrophomonas maltophilia stimulation. AB - Some antibiotics have been shown to modify the host immune response. Infection with Stenotrophomonas maltophilia, is often difficult to treat due to multiresistance to antibiotics. The authors examined the effect of four commonly used antimicrobial agents (ciprofloxacin, ceftazidime, cotrimoxazole and piperacillin-tazobactam) on tumour necrosis factor alpha (TNF alpha) production by human peripheral blood mononuclear cells (PBMC) stimulated with heat-killed S maltophilia. Cotrimoxazole was the only antibiotic that suppressed TNFa secretion at clinically achievable concentrations. This may explain its use with good effect in the treatment of S maltophilia infections. However at supratherapeutic concentrations, ceftazidime and ciprofloxacin, but not piperacillin-tazobactam, also inhibited significantly the production of TNF alpha. Cotrimoxazole, in addition to its antimicrobial effect against S maltophilia, has an immunomodulatory effect on peripheral blood mononuclear cells stimulated by S maltophilia. PMID- 17087096 TI - Skin test reactivity to aeroallergens in Jamaicans: relationship to asthma. AB - BACKGROUND: Asthma causes significant morbidity and mortality in the developing world. It is thus important to identify modifiable risk factors. OBJECTIVES: To undertake a cross-sectional study to determine the prevalence of skin test reactivity to aeroallergens in Jamaican children and adults and the relationship of the diagnosis of asthma to the pattern of skin test positivity. METHODS: One hundred and sixty subjects without the sickle cell gene (genotype AA), eighty adults and eighty children, were recruited. Skin testing to seven aeroallergens was undertaken (atopy being diagnosed if there were at least one positive reaction). Asthma status was determined by a questionnaire and/or medical records. RESULTS: Twenty-seven (34%) of the children and forty-one (51%) of the adults were skin test positive to at least one aeroallergen. The most common positive responses in both age groups were to Dermatophagoides farinae, Dermatophagoides pteronyssinus and cockroach mix-(German and American). All adult asthmatics with current symptoms reacted to cockroach allergen. CONCLUSIONS: Appropriate steps to reduce cockroaches and cockroach sensitization might positively impact on asthma morbidity in Jamaica. PMID- 17087097 TI - A Caribbean perspective on Barrett's oesophagus. AB - The aim of this study is to evaluate the prevalence of Barrett's oesophagus at the University Hospital of the West Indies which serves a population predominantly of African descent. In order to maximize accession of all cases in this preliminary study, the broad definition of columnar metaplasia of the lower oesophagus was utilized. Nevertheless, of 18 cases identified in the 21-year period, 14/17 cases (82%) available for review had specialized columnar epithelium diagnostic of Barrett's oesophagus. There was male predominance 5:1, the mean age was 53.8 years and there were two associated adenocarcinomas. These data confirm the occurrence but low prevalence of Barrett's oesophagus in this population but with a disease pattern that is consistent with that reported in other populations, warranting systematic evaluation of the gastro-oesophageal junction even in a Caribbean population. PMID- 17087098 TI - Mental well-being of doctors and nurses in two hospitals in Kingston, Jamaica. AB - Hospital work involves some of the most stressful situations found in any workplace. Furthermore, hospital workers may be affected by non-work-related stress such as family responsibilities and financial difficulties, leading to impaired mental well-being and suboptimal performance. The aim of this study was to assess the level of general mental well-being among doctors and nurses from two hospitals in Kingston, Jamaica. A total of 212 doctors and nurses at the Kingston Public Hospital and the University Hospital of the West Indies were studied yielding a participation rate of 83.1%. A self-administered questionnaire was used to gather social and biomedical data and the General Health Questionnaire 30 (GHQ 30) used to determine general mental well-being. Probable caseness was defined as a GHQ 30 score > 5. Focus group discussions were also held with staff at both hospitals. A total of 27.4% of the study population met the GHQ-30 criteria (caseness) defining them as probable cases of mental distress. Cases and non-cases were not different in age, gender or hospital of employment. However, caseness was associated with years of professional experience, work-related and non-work-related stress, serious financial difficulties and fears of coming to work. Significant predictors of increased risk of caseness were fear of coming to work (OR 3.06; CI 1.40, 6.70); professional experience in excess of five-years and high non-work-related stress. High work-related stress was associated with reduced risk of being classified a case, suggesting that work may have been therapeutic. Focus group discussions suggested that non-work stress was related to financial difficulties, commuting and child care, especially among nurses. Intervention to improve general mental well-being should be targeted at new employees and should address child care, commuting and financial management. PMID- 17087099 TI - The frequency of smoking, quitting and socio-demographic characteristics of physicians of a medical faculty. AB - BACKGROUND: The aim of this study was to determine the epidemiology of smoking among physicians and to describe their socio-demographic characteristics. SUBJECTS AND METHODS: All professors, associate professors, assistant professors, lecturers and research assistants working for the Faculty of Medicine of Meram, University of Selcuk, were included in the study. Of the 500 subjects, 363 (72.6%) completed the anonymous questionnaire of the survey satisfactorily. The smoking status, age at smoking initiation, daily cigarette consumption and association between current smoking and socio-demographic variables, family characteristics and body mass index were examined. The SPSS software was used to tabulate the data. The chi-square test was used for statistical analysis. RESULTS: The mean age was 34.2 +/- 9.1 years. The rate of ever-smokers was 28.7% (n = 104). The rates of smoking in females and males were 13.4% (n = 15) and 35.6% (n = 89), respectively (p = 0.000). The rate of ex-smokers was 9.9% (n = 36). The rate of never-smokers was 61.4% (n = 223). The mean age at starting smoking was 21.7 +/- 4.9 years. The quit ratio was 25.7% (36/140) [23.3% (27/116) for men and 37.5% (9/24) for women]. Fagerstrom score was averaged 2.8 +/- 2.7 among all smokers. The rate of smoking in the families (p = 0.003) and among close friends (p = 0.000) of the smoker-group was higher than in non-smokers. CONCLUSION: In Turkey, prevalence of tobacco smoking is high among physicians though lower than the general community. This may compromise their role in tobacco control unless they quit smoking. PMID- 17087100 TI - Neonatal sepsis in very low birthweight infants at the University Hospital of the West indies. AB - A retrospective review was conducted on the charts of all very low birthweight (VLBW) infants with culture proven sepsis admitted to the neonatal unit of the University Hospital of the West Indies (UHWI) during the period January 1, 1995 to December 31, 2000. During the study period, 22 VLBW infants were admitted to the neonatal unit with culture proven sepsis, 16 (73%) survived and 6 (27%) died As birthweight and gestational age increased, outcome improved There was no difference in survival based on age at presentation. Neonates with early onset disease had a significantly longer mean duration of rupture of membranes than those with late onset disease (p = 0.009) and babies with late onset disease had a significantly lower mean Hb level than those with early onset disease (p = 0.000). Predominant isolates were Klebsiella sp (10, 37%), Streptococcus Group D (4, 15%), Escherichia coli (3, 11%) and Group B Streptococcus (3, 11%). Klebsiella sp accounted for 8/13 (62%) of late onset infections. Complications included anaemia, thrombocytopenia, bleeding and multi-organ failure. Strategies aimed at prevention, such as limiting the excessive use of broad-spectrum empiric antibiotics and the periodic review and continuous reinforcement of infection control policies will help decrease the mortality and morbidity associated with nosocomial infection in the VLBW infant. PMID- 17087101 TI - Epidemiology of coagulase-negative Staphylococci isolated from clinical blood specimens at the University Hospital of the West Indies. AB - The prevalence and significance of coagulase negative staphylococci (CoNS) isolated from blood cultures at the University Hospital of the West Indies (UHWI) during a six-month period were investigated. Standard and automated microbiological procedures were used to process 3001 blood culture specimens received from 2363 patients and 658 (21.9%) of the blood cultures yielded 854 bacterial isolates. The highest prevalence of positive blood cultures (60%) and the lowest prevalence of blood isolates of CoNS (12%) were found in the intensive care unit (ICU). The blood isolates of CoNS were most frequent in the surgical wards (13%) and lowest in obstetrics and gynaecology (2%). High rates of resistance to methicillin, other anti-staphylococcal penicillins, and cephalosporins used in the treatment of CoNS were observed All blood isolates of CoNS (100%) were susceptible to vancomycin. In conclusion, the results show that coagulase-negative staphylococci are the most prevalent bacterial isolates in blood cultures at the UHWI occurring mostly as contaminants. The practice of proper venepuncture and hand-washing techniques by medical staff are recommended to facilitate appropriate antibiotic usage. PMID- 17087102 TI - Coagulase negative Staphylococci from blood cultures: contaminants or pathogens? AB - Detailed clinical data, underlying conditions, inflammatory indices and microbiological parameters in 60 patients who had pure growth of coagulase negative staphylococci from their blood culture specimens at the University Hospital of the West Indies, Jamaica, were analyzed and the clinical significance of the isolates ascertained using standard criteria. This study was undertaken between April and September 2003. The isolates were true pathogens of bloodstream infections in only 5 of the 60 patients (8.4%). In the vast majority ie 44 of 60 (73.3%) they were mere blood culture contaminants and in 11 (18.3%), the clinical significance could not be ascertained Fifteen of the 44 patients (34%) with contaminating coagulase negative staphylococci were treated with specific anti staphylococcal antibiotics; 5 (11.4%) with vancomycin. Although there has been a relative increase of coagulase negative staphylococcal infections including bloodstream infections in recent years, the organisms still remain the most common contaminants in blood cultures. Over 70% of isolates were contaminants in this study which is similar to that in a number of such studies in other parts of the world The findings underline the need for careful evaluation of coagulase negative staphylococci isolated from blood cultures before instituting therapy to avoid unnecessary use of antibiotics, especially vancomycin, and the consequent increase of antibiotic resistance in hospitals. PMID- 17087103 TI - Nasal carriage of methicillin-resistant and methicillin-susceptible Staphylococcus aureus in nursing home residents in Bolu, Turkey. AB - BACKGROUND: This study aimed (a) to provide information on methicillin-resistant (MRSA) and methicillin-susceptible (MSSA) Staphylococcus aureus colonization ratio in residents of nursing homes; (b) to determine the effect of hand-washing education given to nursing home residents and employees on nasal carriage of Staphylococcus aureus (NCSA) and (c) to obtain probable risk factors for MRSA colonization of residents in two nursing homes. METHODS: Seventy-nine volunteers (24 females and 55 males) from Bolu nursing homes were included in the study. Nasal samples were taken with sterile swabs from the anterior nares. Staphylococcus aureus strains were identified using classical methods and genotyping of methicillin resistant strains was done using Arbitrary Primed PCR (AP-PCR). Antibiotic susceptibilities were determined by disk diffusion methods according to NCCLS standards. After first nasal samples were taken, all employees and residents of nursing homes were educated about the methods of hand hygiene over two days. RESULTS: With hand-washing education, the decrease of NCSA rate (initially 43%; after education, 21%) was significant (p < 0.05) while decrease of MRSA carriage (initially 5%, after education 1%) was not (p > 0.05). MRSA carriage was significantly correlated with presence of skin lesions, prior hospitalization within the last six months, and antibiotics usage within the last six months. AP PCR results suggested that residents' carriage of MRSA was the result of the same source. CONCLUSION: MSSA and MRSA colonization rates were found to be 38% and 5% in nursing homes, respectively. These ratios can decrease with simple precautions like hand-washing after a short education period PMID- 17087104 TI - Nosocomial urinary tract infections: micro-organisms, antibiotic sensitivities and risk factors. AB - OBJECTIVE: Although urinary tract infections (UTIs) are the most common hospital acquired infections, the epidemiology of these UTIs is not well defined in Turkey. The aim of this surveillance study was to determine micro-organisms responsible for UTI, their antibiotic sensitivities and to describe the incidence and risk factors of nosocomial urinary tract infections (NUTI). SUBJECTS AND METHODS: This was a prospective surveillance study including cases of NUTI in intensive care units and various inpatient clinics. This study was carried out between November 2000 and January 2002. The following information was recorded: patients' age, gender, type of infection (hospital-acquired), presence of urinary catheter, intensive care unit admission, duration of hospital stay, type of organisms isolated and their antimicrobial susceptibility. The diagnosis of NUTI was based on criteria established by the Centers for Disease Control, Atlanta. Mini Api and conventional culture methods were used to determine the causative agents. The agents were isolated on eosin methylene blue agar and 5% sheep blood agar Statistical analyses of data were by chi-square test and logistic regression. RESULTS: In this study, 618 (2.1%) nosocomial infections (NIs) were determined in 29,778 patients, and 178 of these infections were NUTI (28.8%, 178/618). The mean age of NUTI patients was 61.0 +/- 19.4 years (0-91 years) and 82 NUTI patients (46.1%) were male and 96 (53.9%) were female. The most frequently isolated micro-organism was Escherichia coli (31.4%) followed by Candida spp (21.3%), Klebsiella spp (10.6%) and Enterococcus spp (6.9%). Compared to the rate of other NIs, the rate of NUTI increased by 1.011 times per year of age, by 2.052 times in females and by 3.83 times in patients with urinary catheters (p < 0.05). The most effective antibiotics against Gram-negative bacteria were found to be imipenem and meropenem. CONCLUSIONS: Important factors to prevent NUTI are to avoid unnecessary urethral catheterization, to choose narrow spectrum antibiotics according to antibiotic sensitivities, to investigate regularly the causative micro-organisms and their resistance patterns and to update the treatment protocols. PMID- 17087105 TI - Resistance in clinical isolates of Enterococcusfaecalis encountered at the University Hospital of the West Indies, Jamaica. AB - Enterococcus faecalis isolates were examined by an automated identification and susceptibility system. Almost all of the 97 isolates were ampicillin susceptible (n = 86) and tetracycline resistant (n = 89). All were nitrofurantoin susceptible. About a third of isolates showed high level resistance to the aminoglycosides streptomicin and gentamicin and this was usually associated with ciprofloxacin resistance (n = 34). Seven isolates were vancomycin resistant, including one that was ampicillin resistant. Most forms of resistance described elsewhere were found PMID- 17087106 TI - Choline: are our university students eating enough? AB - Choline is an essential nutrient; dietary deficiency of choline is associated with impaired liver function, elevated blood concentrations of alanine aminotransferase, creatinine phosphokinase and homocysteine. There is also depletion of acetylcholine concentration in the brain, leading to deficit in memory function. The authors examined the dietary intake of choline in groups of students at the Mona Campus of the University of the West Indies. Sixty-two medical students (first and second years) and biochemistry students (final year) were recruited They were asked to (including amounts) record all foods and drinks consumed for three days (two weekdays and one weekend day). The sheets were collected and the amount of choline and betaine (a metabolite of choline) consumed were calculated Dietary intake of folate was also evaluated. The analysis revealed that 86.2% of the females and 90.9% of the males reported diets that delivered less daily choline than the adequate intake quoted by the Institute of Medicine of the National Academy of Sciences, USA (425-550 mg/day). The betaine consumption ranged between 25 to 620 mg/day (no adequate intake documented) and the folate consumed was more than the recommended daily allowance of folate (180-200 microg/day). The dietary intake of choline in the majority of students is below adequate intake. Although folate also serves similar functions to choline, it is unlikely that it can substitute for choline in all physiological aspects and therefore the implications of low dietary choline need further investigation. PMID- 17087107 TI - A fatal case of community acquired methicillin resistant Staphylococcus aureus brain abscess in a previously healthy adolescent. AB - A 16-year-old adolescent presented with fever, lethargy and vomiting associated with mild dehydration. This was followed less than 24-hours later by loss of consciousness, seizures and clinical brain death. She had no prior medical illness, no hospital or frequent antibiotic exposure. There was no evidence of soft tissue or skin infection. Management included intravenous fluids, ampicillin and cefotaxime parenterally, dexamethasone, endotracheal intubation and mechanical ventilation. Her neurologic examination remained unchanged with areflexia, flaccid paralysis and fixed pupils. Post-mortem examination revealed an eight-centimetre right fronto-parietal lobe brain abscess. Cultures were positive for methicillin resistant Staphylococcus aureus. Although formerly a nosocomial pathogen affecting debilitated patients in the hospital setting, S aureus that is methicillin resistant is emerging as a community acquired pathogen affecting previously well patients. PMID- 17087108 TI - Some bio-medical mechanisms in athletic prowess. AB - Variations in somato-genetic patterns in muscle-fibre biology, biochemical metabolic pathways and pulmonary physiology are hypothesized to have been concentrated by natural selection over the centuries in the Afrocentric peoples displaced from West Africa to the New World. These phenotypic and genotypic characteristics are attributed to provide the athletic prowess so well documented in African-Americans. Not the least of coincidence seems to be the influence of the compensatory mechanisms on oxygen transport and its availability to the tissues, in response to the sickle cell gene. The reduced availability coupled with reduced myoglobin in the preponderant fast-twitch muscle fibres which are adapted for rapid energy (ATP) regeneration, all give a NET outcome of muscle anatomical and biochemical advantages which support outstanding performances in athleticism. PMID- 17087109 TI - Image and diagnosis. Cutaneous larva migrans (CLM). PMID- 17087110 TI - [The correlation of PDGF and TNF-alpha and HA and nasal mucosa healing after ESS]. AB - OBJECTIVE: To investigate the relation of healing outcomes and three factors in nasal fluid before and after endoscopic sinus surgery (ESS), including platelet derived growth factor (PDGF), tumour necrosis factor-alpha (TNF-alpha) and hyaluronic acid (HA). METHOD: We compared the amounts of PDGF, TNF-alpha, HA among the three stages in 22 patients operated for nasal polyposis. The concentrations of PDGF, TNF-alpha, HA were compared before and after operation between good healing and poor healing at 12th week, and gender, age, body mass, preoperative stage were also compared. Finally, we performed a multiple logistic regression model to assess whether the healing quality was associated with gender, age, body mass, preoperative stage, and concentrations of PDGF, TNF alpha, HA before ESS. RESULT: (1) A significant difference in the concentrations of PDGF and HA could be demonstrated between the first and second stage, or between the first and third stage. But there was no difference in TNF-alpha among three stages. (2) The healing quality after ESS was significantly and independently correlated to the age of patient and preoperative PDGF concentrations in nasal secretions, the younger, the better healing. The lower PDGF concentration, the better healing, and vice versa. CONCLUSION: During the wound healing of nasal mucosa, the levels of PDGF, TNF-alpha, HA are different at each postoperative stage. Age and PDGF concentrations preoperatively are suitable factors to predict the healing quality after ESS. PMID- 17087111 TI - [A binding study of the gross and endoscopic anatomy of optic canal]. AB - OBJECTIVE: To provide transnasal endoscopic optic nerve decompression with the anatomic reference. METHOD: Ten (20 sides) damp cadaveric heads of Chinese adults were cut sagittally along the midlines. The distance and angle between optic canal and anterior nasal spine were measured by ruler and protractor. Five (10 sides) damp cadaveric heads (including 2 children's) were undergone endoscopic dissection, and the optic canal and its related structure were observed. RESULT: From the study of gross anatomy, the specific structure of optic canal and internal carotid artery were found. The mean length of the medial wall of optic canal was (9.12 +/- 1.89) mm. The mean diameter of the optic canal orbital aperture was (4.12 +/- 0.53) mm. The distances from the midpoint of the medial wall of optic canal orbital aperture to anterior nasal spine was (61.22 +/- 6.23) mm, and the angle from the midpoint of the medial wall of optic canal orbital aperture to anterior nasal spine was (45.3 +/- 4.5) degrees. From the study of endoscopic anatomy, optic canal, looking like a light-reflective band, was alone optic canal orbital aperture posteriorly. Obvious prominence could be found in 7 sides (70%), while not been seen clearly in 3 sides (30%). CONCLUSION: The binding study of the gross and endoscopic anatomy of optic canal help us to recognize optic canal accurately under nasal endoscope, which can improve the veracity of transnasal endoscopic optic nerve decompression. PMID- 17087112 TI - [The roles of three peptides in the pathogenesis of vasomotor rhinitis]. AB - OBJECTIVE: To explore the possible roles of three neuropeptides in the pathogenesis of vasomotor rhinitis by studying the expression and distribution of neuropeptides in the mucosa of vasomotor rhinitis, such as Substance P(SP), calcitonin gene-related peptide (CGRP) and vasoactive intestinal peptide(VIP). METHOD: The mucosa specimens of thirty vasomotor rhinitis patients who had typical symptoms and signs were selected randomly as the experiment group and were divided into two subgroups depending on if received treatment or not. While the normal middle turbinate mucosa specimens of nine cases were selected as the control group. The expression and distribution of neuropeptides were examined by immunohistochemical SP method and computer image disposing and analyzing system. RESULT: The terminals of SP, CGRP and VIP in the treated experiment group wer and untreated experiment group were markedly increased in density of immunostaining compared to the control group,and the difference is significant (P < 0.01). CONCLUSION: Neuropeptides, such as SP, CGRP and VIP, may play important roles in the pathophysiological mechanism of vasomotor rhinitis. PMID- 17087113 TI - [Expression of transforming growth factor beta receptor I and II in chronic rhinosinusitis, nasal polyps and normal nasal mucosa tissues]. AB - OBJECTIVE: To explore the expression of transforming growth factor beta receptor(TGFpR) I, II in the chronic rhinosinusitis, nasal polyps and normal nasal mucosa tissues. METHOD: The protein expression of the TGFbetaR I and TGFbetaR II were determined by means of immunohistochemistry in chronic rhinosinusitis tissues from 25 patients, nasal polyps tissues from 21 patients and inferior turbinate mucosa tissues from 17 patients with deviation of nasal septum. RESULT: (1) In chronic rhinosinusitis and nasal polyps tissues, compared with controls, the expression of TGFbetaR I and TGFbetaR II protein was increased significantly (P < 0.01). (2) The expression of TGFbetaR I and TGFbetaR II protein in nasal polyps tissues was significant increased than that in chronic rhinosinusitis tissues (P < 0.01). CONCLUSION: The different expression level and pattern of TGFbetaR I and TGFbetaR II in chronic rhinosinusitis and nasal polyps tissues indicates TGFbetaR may play a different role in the pathogenesis chronic rhinosinusitis and nasal polyps. PMID- 17087114 TI - [Foreign body in tracheobronchial diagnosed as pneumonia from children]. AB - OBJECTIVE: To improve the effects of diagnosis and therapy in patients with tracheobronchial foreign body who were diagnosed as pneumonia and to summarize the experience. METHOD: Foreign bodies in the trachea and bronchi in one hundred and seventy-two children were removed in our hospital from 1995. 2 to 2005. 2, of them 67 cases were diagnosed as pneumonia. RESULT: Our experience showed that the foreign body in tracheobronchial had an increasing tendency in recent years, and it had a descendant tendency that it be diagnosed as pneumonia. The time of the patients suffered from foreign bodies was from 5 days to 8 months, and the children under 2 years accounted for 80.6%. The most common foreign body was peanuts, accounting for 70.4%. All the foreign bodies were removed by rigid bronchoscopy. The successful rate of removal at first time was 63 cases, accounting for 94%. The reason of failure at first time was anesthesia (2 cases). There was no case of death. No cases underwent tracheotomy. CONCLUSION: The foreign body in tracheobronchial can be easily diagnosed as pneumonia. We should reduce the rate of erroneous diagnosis. Rigid bronchoscopy for removing foreign bodies under general anesthesia is still the main measure at present. PMID- 17087115 TI - [Comparative study of uptake and washout of 99mTcn(NOEt)2 with 99mTc-MIBI in human nasopharyngeal carcinoma cell line CNE]. AB - OBJECTIVE: To determine the cellular kinetics of 99mTc-bis {N-ethoxy-N-ethyl dithiocarbamato-nitrido [N(NOEt)2]} in human nasopharyngeal carcinoma CNE cells line and to compare it with the cellular kinetics of 99mTc methoxyisobutylisonitrile (MIBI) in the same conditions; to define the possible clinical application value of 99mTcN(NOEt)2 in nasopharyngeal carcinoma imaging. METHOD: 99mTcN(NOEt)2 or 99mTc-MIBI were incubated with CNE cells at 37degrees C, 22 degrees C, at different time points after incubation, the uptake and washout characteristics of radiotracers in CNE cells were investigated and compared. RESULT: The maximum uptake ratio of 99mTcN(NOEt)2 in CNE cells was 43.15%, that of 99mTc-MIBI was 11.08% (P < 0.01). The uptake of 99mTcN(NOEt)2 after 5 min incubation in CNE cells was 63% of total uptake, uptake of 99mTc-MIBI was 48% of total uptake (P < 0.05); 1 h after removal of the unbound tracer, 63.92% of the total uptake of 99TcN(NOEt)2 was retained in the CNE cells, 53. 97% of 99mTc-MIBI was retained (P < 0.05); not like 99mTc-MIBI, the cellular kinetics of 99mTcN (NOEt)2 were similar at 37 degrees C and 22 degrees C ( P > 0.05). CONCLUSION: 99mTcN(NOEt)2 may be a gamma imaging agent better than 99mTc-MIBI for nasopharyngeal carcinoma imaging. 99mTcN(NOEt)2 is a potential human nasopharyngeal carcinoma imaging agent. PMID- 17087116 TI - [An experimental study on the regulation of expression of Th1/Th2 cytokines by allergen vaccine atomization inhalation in patients with asthma]. AB - OBJECTIVE: To explore the role of specific immunotherapy treatment (SIT) with allergen vaccine (AV) atomization inhalation in regulation of the expression of Th1/Th1 cytokines (IL-2 and IL-4) by T lymphocyte and serum IgE level in asthmas. METHOD: Ninety cases with asthma in remission were randomly divided into three groups and each group contained thirty patients who received SIT respectively with atomization inhalation (A group), sublingual contained (B group) and hypodermic injection (C group). 32 healthy subjects were as control group. Before and after treatment, the expression of IL-2 and IL-4, total IgE level were measured by radio-immune. The three SIT treatments were evaluated. RESULT: Before SIT treatment, compared with the normal control group, the patients' serum level of Th2 cytokines IL-4 and total IgE had significantly increased, the level of Th1 cytokines IL-2 had significantly reduced. There were statistical significance between the asthma and normal control (t = 4.869, 5.598, 10.009, P < 0.01). After 12 months SIT treatment, the patients' serum IL-2 showed increased, the serum IL 4 and total IgE showed reduced, the difference was statistical significant (t = 7.303, 2.494, 2.225, P < 0.05). There was good positive correlation between the level of IL-4 and the total IgE (r = 0.264, P < 0.01), there was good negative correlation between the level of IL-2 and the total IgE (r = -0.369, P < 0.001), and there was good negative correlation between the level of IL-2 and IL-4 (r = 0.277, P < 0.01). The effects among A, B, C groups were similar (all were over 86%), without statistical significance (chi2 = 0.055, P > 0.05). CONCLUSION: The specific response to antigen may be transferred from Th2 to Th1 by SIT with AV atomization inhalation, and mediated between balance of Th1/Th2 in patients with asthma. This method is thought a direct influence on the target organ. It is safe, simple and valid and can be easily accepted by sufferers. PMID- 17087117 TI - [Effect of radiation on nasal mucosa and the microvascular casting of guinea pig]. AB - OBJECTIVE: To study the histopathologic and microvascular changes in nasal mucosa of guinea pigs following a series of irradiation. METHOD: A total of 84 healthy guinea pig were divided into two groups randomly: the irradiation group (n=72) and the control group (n=12). The animals of the irradiation group were exposed nasally to linear accelerator X-rays at 5.0 Gy once a week for three weeks. At 0.5, 1, 2, 3, 5, 6 month after X-irradiation 12 animals from each group were sacrificed at each time point. The animals of control group did not be irradiated and were sacrificed after 6 month. Their middle nasal turbinate mucosa of 6 animals from each group was observed under optical microscope and scanning electron microscope and the another 6 animals was made microvascular corrosion casts. RESULT: In the irradiation group, the early histopathological transformation was acute inflammatory reaction, nasal mucosa tissues congesting, partial nasal mucosa necrosis or shedding. The inflammation cells infiltrated into the submucosa. The acute inflammatory reaction declined as the time went by. Then the nasal mucosa repaired step by step. But parts of the area were squamous epithelium instead of the normal ciliary structure. Through scanning electron microscope, we found the capillary of nasal mucosa expanded and congested, the vascular endothelial cell swelled. As the time went by, the capillary became deformation and tore and the microvascular networks broke off. All changes which led to capillary density descended. Although partial area had new capillary, but the structure was disordered and had no microcirculation function. CONCLUSION: The nasal mucosa injury, the squamous metaplasia and the microvascular networks broke off can form a pathological basis of dysfunction after irradiation. PMID- 17087118 TI - Peripheral and central mechanisms of pain generation. AB - Pain research has uncovered important neuronal mechanisms that underlie clinically relevant pain states such as inflammatory and neuropathic pain. Importantly, both the peripheral and the central nociceptive system contribute significantly to the generation of pain upon inflammation and nerve injury. Peripheral nociceptors are sensitized during inflammation, and peripheral nerve fibres develop ectopic discharges upon nerve injury or disease. As a consequence a complex neuronal response is evoked in the spinal cord where neurons become hyperexcitable, and a new balance is set between excitation and inhibition. The spinal processes are significantly influenced by brain stem circuits that inhibit or facilitate spinal nociceptive processing. Numerous mechanisms are involved in peripheral and central nociceptive processes including rapid functional changes of signalling and long-term regulatory changes such as up-regulation of mediator/receptor systems. Conscious pain is generated by thalamocortical networks that produce both sensory discriminative and affective components of the pain response. PMID- 17087119 TI - Opioids. AB - Opioids are the most effective and widely used drugs in the treatment of severe pain. They act through G protein-coupled receptors. Four families of endogenous ligands (opioid peptides) are known. The standard exogenous opioid analgesic is morphine. Opioid agonists can activate central and peripheral opioid receptors. Three classes of opioid receptors (mu, delta, kappa) have been identified. Multiple pathways ofopioid receptor signaling (e.g., G(i/o) coupling, cAMP inhibition, Ca++ channel inhibition) have been described. The differential regulation of effectors, preclinical pharmacology, clinical applications, and side effects will be reviewed in this chapter. PMID- 17087120 TI - Antipyretic analgesics: nonsteroidal antiinflammatory drugs, selective COX-2 inhibitors, paracetamol and pyrazolinones. AB - Antipyretic analgesics are a group of heterogeneous substances including acidic (nonsteroidal antiinflammatory drugs, NSAIDs) and nonacidic (paracetamol, pyrazolinones) drugs. Moreover, various selective cyclooxygenase-2 (COX-2) inhibitors with improved gastrointestinal tolerability as compared with conventional NSAIDs have been established for symptomatic pain treatment in recent years. The present review summarizes the pharmacology of all of these drugs with particular emphasis on their rational use based on the diverse pharmacokinetic characteristics and adverse drug reaction profiles. Referring to the current debate, potential mechanisms underlying cardiovascular side effects associated with long-term use of COX inhibitors are discussed. PMID- 17087121 TI - Local anesthetics. AB - Local anesthetics are used broadly to prevent or reverse acute pain and treat symptoms of chronic pain. This chapter, on the analgesic aspects of local anesthetics, reviews their broad actions that affect many different molecular targets and disrupt their functions in pain processing. Application of local anesthetics to peripheral nerve primarily results in the blockade of propagating action potentials, through their inhibition of voltage-gated sodium channels. Such inhibition results from drug binding at a site in the channel's inner pore, accessible from the cytoplasmic opening. Binding of drug molecules to these channels depends on their conformation, with the drugs generally having a higher affinity for the open and inactivated channel states that are induced by membrane depolarization. As a result, the effective potency of these drugs for blocking impulses increases during high-frequency repetitive firing and also under slow depolarization, such as occurs at a region of nerve injury, which is often the locus for generation of abnormal, pain-related ectopic impulses. At distal and central terminals the inhibition of voltage-gated calcium channels by local anesthetics will suppress neurogenic inflammation and the release of neurotransmitters. Actions on receptors that contribute to nociceptive transduction, such as TRPV1 and the bradykinin B2 receptor, provide an independent mode of analgesia. In the spinal cord, where local anesthetics are present during epidural or intrathecal anesthesia, inhibition of inotropic receptors, such as those for glutamate, by local anesthetics further interferes with neuronal transmission. Activation of spinal cord mitogen-activated protein (MAP) kinases, which are essential for the hyperalgesia following injury or incision and occur in both neurons and glia, is inhibited by spinal local anesthetics. Many G protein-coupled receptors are susceptible to local anesthetics, with particular sensitivity of those coupled via the Gq alpha subunit. Local anesthetics are also infused intravenously to yield plasma concentrations far below those that block normal action potentials, yet that are frequently effective at reversing neuropathic pain. Thus, local anesthetics modify a variety of neuronal membrane channels and receptors, leading to what is probably a synergistic mixture of analgesic mechanisms to achieve effective clinical analgesia. PMID- 17087122 TI - Serotonin receptor ligands: treatments of acute migraine and cluster headache. AB - Fuelled by the development of the serotonin 5-HT(1B/1D) receptor agonists, the triptans, the last 15 years has seen an explosion of interest in the treatment of acute migraine and cluster headache. Sumatriptan was the first of these agonists, and it launched a wave of therapeutic advances. These medicines are effective and safe. Triptans were developed as cranial vasoconstrictors to mimic the desirable effects of serotonin, while avoiding its side-effects. It has subsequently been shown that the triptans' major action is neuronal, with both peripheral and central trigeminal inhibitory effects, as well as actions in the thalamus and at central modulatory sites, such as the periaqueductal grey matter. Further refinements may be possible as the 5-HT(1D) and 5-HT(1F) receptor agonists are explored. Serotonin receptor pharmacology has contributed much to the better management of patients with primary headache disorders. PMID- 17087123 TI - Anti-convulsants and anti-depressants. AB - Damage to a nerve should only lead to sensory loss. While this is common, the incidence of spontaneous pain, allodynia and hyperalgesia indicate marked changes in the nervous system that are possible compensations for the loss of normal function that arises from the sensory loss. Neuropathic pain arises from changes in the damaged nerve which then alter function in the spinal cord and the brain and lead to plasticity in areas adjacent to those directly influenced by the neuropathy. The peripheral changes drive central compensations so that the mechanisms involved are multiple and located at a number of sites. Nerve damage increases the excitability of both the damaged and undamaged nerve fibres, neuromas and the cell bodies in the dorsal root ganglion. These peripheral changes are substrates for the ongoing pain and the efficacy of excitability blockers such as carbamazepine, lamotrigine and mexiletine, all anti-convulsants. A better understanding of ion channels at the sites of injury has shown important roles of particular sodium, potassium and calcium channels in the genesis of neuropathic pain. Within the spinal cord, increases in the activity of calcium channels and the receptors for glutamate, especially the N-methyl-D-aspartate (NMDA) receptor, trigger wind-up and central hyperexcitability. Increases in transmitter release, neuronal excitability and receptive field size result from the damage to the peripheral nerves. Ketamine and gabapentin/pregabalin, again with anti-convulsant activity, may interact with these mechanisms. Ketamine acts on central spinal mechanisms of excitability whereas gabapentin acts on a subunit of calcium channels that is responsible for the release of pain transmitters into the spinal cord. In addition to these spinal mechanisms of hyperexcitability, spinal cells participate in a spinal-supraspinal loop that involves parts of the brain involved in affective responses to pain but also engages descending excitatory and inhibitory systems that use the monoamines. These pathways become more active after nerve injury and are the site of action of anti-depressants. This chapter reviews the evidence and mechanisms of drugs, both anti-depressants and anti-convulsants, that are believed to be effective in pain control, with a major emphasis on the neuropathic state. PMID- 17087124 TI - Neuropeptide and kinin antagonists. AB - Neuropeptides and kinins are important messengers in the nervous system and--on the basis of their anatomical localisation and the effects produced when the substances themselves are administered, to animals or to human subjects-a significant number of them have been suggested to have a role in pain and inflammation. Experiments in gene deletion (knock-out or null mutant) mice and parallel experiments with pharmacological receptor antagonists in a variety of species have strengthened the evidence that a number of peptides, notably substance P and calcitonin gene-related peptide (CGRP), and the kinins have a pathophysiological role in nociception. Clinical studies with non-peptide pharmacological antagonists are now in progress to determine if blocking the action of these peptides might have utility in the treatment of pain. PMID- 17087125 TI - Glutamate receptor ligands. AB - Glutamate acts through a variety of receptors to modulate neurotransmission and neuronal excitability. Glutamate plays a critical role in neuroplasticity as well as in nervous system dysfunctions and disorders. Hyperfunction or dysfunction of glutamatergic neurotransmission also represents a key mechanism of pain-related plastic changes in the central and peripheral nervous system. This chapter will review the classification of glutamate receptors and their role in peripheral and central nociceptive processing. Evidence from preclinical pain models and clinical studies for the therapeutic value of certain glutamate receptor ligands will be discussed. PMID- 17087126 TI - Adrenergic and cholinergic compounds. AB - Adrenergic and cholinergic signalling contributes significantly to the endogenous antinociceptive system. Exogenous alpha 2 adrenergic agonists have a well established analgesic profile; however, recent investigations suggest that this class of agents is underused, and herein we highlight the potential for both current application and future development of these agents. Nicotinic and muscarinic cholinergic ligands represent a novel class of agents with much promise for the management of problematic pain. In this chapter we review advances in both preclinical and clinical arenas and highlight potential avenues for further research. PMID- 17087127 TI - Cannabinoids and pain. AB - Convincing evidence from preclinical studies demonstrates that cannabinoids can reduce pain responses in a range of inflammatory and neuropathic pain models. The anatomical and functional data reveal cannabinoid receptor-mediated analgesic actions operating at sites concerned with the transmission and processing of nociceptive signals in brain, spinal cord and the periphery. The precise signalling mechanisms by which cannabinoids produce analgesic effects at these sites remain unclear; however, significant clues point to cannabinoid modulation of the functions of neurone and immune cells that mediate nociceptive and inflammatory responses. Intracellular signalling mechanisms engaged by cannabinoid receptors-like the inhibition of calcium transients and adenylate cyclase, and pre-synaptic modulation of transmitter release-have been demonstrated in some of these cell types and are predicted to play a role in the analgesic effects of cannabinoids. In contrast, the clinical effectiveness of cannabinoids as analgesics is less clear. Progress in this area requires the development of cannabinoids with a more favourable therapeutic index than those currently available for human use, and the testing of their efficacy and side effects in high-quality clinical trials. PMID- 17087128 TI - Adenosine and ATP receptors. AB - Adenosine and ATP, via P1 and P2 receptors respectively, can modulate pain transmission under physiological, inflammatory, and neuropathic pain conditions. Such influences reflect peripheral and central actions and effects on neurons as well as other cell types. In general, adenosine A1 receptors produce inhibitory effects on pain in a number of preclinical models and are a focus of attention. In humans, i.v. infusions of adenosine reduce some aspects of neuropathic pain and can reduce postoperative pain. For P2X receptors, there is a significant body of information indicating that inhibition of P2X3 receptors may be useful for relieving inflammatory and neuropathic pain. More recently, data have begun to emerge implicating P2X4, P2X7 and P2Y receptors in aspects of pain transmission. Both P1 and P2 receptors may represent novel targets for pain relief. PMID- 17087129 TI - Ion channels in analgesia research. AB - The distribution of ion channels in neurons associated with pain pathways is becoming better understood. In particular, we now have insights into the molecular nature of the channels that are activated by tissue-damaging stimuli, as well as the mechanisms by which voltage-gated channels alter the sensitivity of peripheral neurons to change pain thresholds. This chapter details the evidence that individual channels may be associated with particular pain states, and describes genetic approaches to test the possible utility of targeting individual channels to treat pain. PMID- 17087130 TI - Protein kinases as potential targets for the treatment of pathological pain. AB - Pathological pain or clinical pain refers to tissue injury-induced inflammatory pain and nerve injury-induced neuropathic pain and is often chronic. Pathological pain is an expression of neural plasticity that occurs both in the peripheral nervous system (e.g., primary sensory nociceptors), termed peripheral sensitization, and in the central nervous system (e.g., dorsal horn and brain neurons), termed central sensitization. Our insufficient understanding of mechanisms underlying the induction and maintenance of injury-induced neuronal plasticity hinders successful treatment for pathological pain. The human genome encodes 518 protein kinases, representing one of the largest protein families. There is growing interest in developing protein kinase inhibitors for the treatment of a number of diseases. Although protein kinases were not favored as targets for analgesics, studies in the last decade have demonstrated important roles of these kinases in regulating neuronal plasticity and pain sensitization. Multiple protein kinases have been implicated in peripheral and central sensitization following intense noxious stimuli and injuries. In particular, mitogen-activated protein kinases (MAPKs), consisting of extracellular signal regulated kinase (ERK), p38, and c-Jun N-terminal kinase (JNK), are downstream to many kinases and are activated in primary sensory and dorsal horn neurons by nociceptive activity, growth factors and inflammatory mediators, contributing to the induction and maintenance of pain sensitization via posttranslational, translational, and transcriptional regulation. MAPKs are also activated in spinal glial cells (microglia and astrocytes) after injuries, leading to the synthesis of inflammatory mediators/neuroactive substances that act on nociceptive neurons, enhancing and prolonging pain sensitization. Inhibition of multiple kinases has been shown to attenuate inflammatory and neuropathic pain in different animal models. Development of specific inhibitors for protein kinases to target neurons and glial cells will shed light on the development of new therapies for debilitating chronic pain. PMID- 17087131 TI - Placebo and endogenous mechanisms of analgesia. AB - The discovery of the endogenous systems of analgesia has produced a large amount of research aimed at investigating their biochemical and neurophysiological mechanisms and their neuroanatomical localization. Nevertheless, the neurobiological acquisitions on these mechanisms have not been paralleled by behavioural correlates in humans--in other words, by the understanding of when and how these endogenous mechanisms of analgesia are activated. Until recent times one of the most studied behavioural correlates of endogenous analgesia was stress-induced analgesia, in which the activation of endogenous opioid systems is known to be involved. By contrast, today the placebo analgesic effect represents one of the best-described situations in which this endogenous opioid network is naturally activated in humans. Therefore, not only is placebo research helpful towards improving clinical trial design and medical practice, but it also provides us with a better understanding of the endogenous mechanisms of analgesia. PMID- 17087132 TI - Limitations of pharmacotherapy: behavioral approaches to chronic pain. AB - Pharmacotherapy is most appropriate in acute pain, whereas in chronic pain states behavioral approaches or a combination of behavioral treatment and pharmacotherapy is more appropriate. In this chapter we first describe the role of learning and memory as well as other psychological factors in the development of chronic pain and emphasize that chronic pain must viewed as the result of a learning process with resulting central neuroplastic changes. We then describe operant behavioral and cognitive-behavioral treatments as well as biofeedback and relaxation techniques and present innovative treatment procedures aimed at altering central pain memories. We complete the section with a discussion of combined behavioral and pharmacological approaches and an interdisciplinary view. PMID- 17087133 TI - [Determination of grade of urothelial carcinoma according to WHO 2004 classification]. AB - Several classifications of transitional/urothelial cell tumors have been proposed during last few years in order to standardize nomenclature, and criteria for grading and invasion. They also aimed to avoid the term cancer for neoplasms that very rarely invade, recur, and/or cause death of the patient. As a result of these efforts a new WHO classification emerged in the year 2004. Instead of the term transitional, the use of urothelial neoplasms was recommended. In the group of noninvasive urothelial neoplasms, a new category of tumor of low malignant potential was added. Three-tier grading of papillary noninvasive tumor was substituted by low and high-grade category. Criteria for the grades are cited in the classification but are somewhat imprecise and difficult to apply. On the basis of the data from the literature and our own experience, in the transitional period we recommend the use of WHO 1973 simultaneously with the new one. Problems of the identification of lamina propria invasion are still not solved, and therefore the use of additional histochemical and immunohistochemical methods should be recommended in difficult cases. PMID- 17087134 TI - [The importance of Streptococcus bovis systemic infections]. AB - Streptococcus bovis is relatively common cause of bacteremia and endocarditis, especially in older persons, or in patients that have some kind of chronic disease. These infections are frequently connected with malignant, potentially malignant, or benign colorectal neoplasia. Hematogenous dissemination of Streptococcus bovis could result with various clinical manifestations, namely purulent meningitis, brain abscess, osteomyelitis, spondylodiscitis, and many different kinds of infections in AIDS patients are reported. In this report, two patients hospitalized at the Departement of Infectious Diseases at the General Hospital "Dr. Josip Bencevic" in Slavonski Brod are presented. The first patient was addmited to the hospital because of fever of unknown origin, and in his blood cultures, Streptococcus bovis was isolated. He was an older man, who had undergone prostatectomy due prostatic adenoma several years before. The other patient, previously completely healthy younger man, suffered purulent meningitis caused by the same microorganism. Colon endoscopy was performed in both patients and it revealed colon polyps. Histologically, in both cases, those were benign neoplasia. In Croatia, until this report, there have been no other reports about patients suffering systemic Streptococcus bovis infections. At the same time, this report describes Streptococcus bovis purulent meningitis in a previously healthy adult, which is also extremely rare in the medical literature. Since Streptococcus bovis infections are associated with colon carcinoma, it is imperative to perform colonoscopy in each patient suffering infection with this germ, and to consider him as a high risk patient for developing colon cancer. PMID- 17087135 TI - [Secondary breast bilateral reconstruction with myocutaneous pedicled latissimus dorsi flap after primary bilateral skin sparing mastectomy reconstruction with consecutive severe capsular contraction]. AB - We present a patient who was operated due to bilateral breast carcinoma with immediate bilateral breast reconstruction with silicone implants after skin sparing mastectomy in a neighbouring country to Croatia. One year following the operation a severe bilateral capsular contracture was manifested. Due to a large umbilical hernia and lower laparotomy scar it was not possible to reconstruct the breasts with any abdominal free or pedicled flap. We performed bilateral secondary breast reconstruction with latissimus dorsi myocutaneous flap and silicone implants in two stages with good postoperative result. PMID- 17087136 TI - [Lumbar disc herniation. Reposition effect of rotational manipulation: a case report]. AB - In a 38-year old patient affected by acute radicular syndrome magnetic resonance imaging (MRI), diagnosed a large dorso-medial herniation at L4/L5 segment. Upon application of the usual conservative therapy, during 3.5 months of conservative treatment, no clinical improvement was achieved. On the new MRI, made after the completion of physical therapy (the patient was treated for 30 days in a thermal resort), i.e. 40 days subsequent to the first MRI or 3.5 months after the onset of symptoms, no signs of natural regression of the herniation, i.e. changes in the size of herniation, were noted. After unsuccessful conservative treatment, neurosurgeon recommended a surgical procedure which was postponed by the patient for a definite period of time. In the meantime, the patient underwent a manipulative treatment. In the course of the manipulative treatment, a significant subjective and clinical improvement was reached. In the segment affected by the herniation, four rotational manipulations all together were carried out at the intervals often days. On the new MRI made upon the fourth manipulation, i.e. 40 days subsequent to the second MRI, an almost complete regression of the herniation was found which corresponded to the clinical outcome. A conclusion can be drawn that, by rotational manipulation, a reposition effect has been achieved. The patient described in these papers does corroborate the viewpoint of several authors on effectiveness of rotational manipulation in patients suffering from lumbar disc herniation. PMID- 17087137 TI - [The role of endoscopic ultrasound in digestive tract diseases in era of advanced techniques of computed tomography]. AB - During the last 20 years endoscopic ultrasound (EUS) has been established as a highly accurate method in the diagnosis of digestive tract diseases. This method enables analysis of the lesions within the bowel wall, pancreas, and biliary system with utmost precision. Good results have also been achieved in evaluation of regional lymphadenopathy in staging of malignant diseases. Principal limitations of this method originate from its low accesibility and technical complexity which requires well trained sinologist and scientific evaluation of results in order to achieve adequate level of skill. Complications occur rarely with numbers comparable to other endoscopic procedures. With new software and hardware device introduced, computerized tomography (CT) (this refers in the first place to multidetector row CT) has been tremendously upgraded approaching the EUS in diagnostic accuracy in mentioned indications, and overcoming it in detection of distant metastases. Implementation of 3D reconstruction techniques allowed CT to enter the area of classic endoscopy, as in the case of virtual gastroscopy and/or colonoscopy. CT is a noninvasive method, very acceptable to majority of patients, and therefore popular and more prevalent compared to EUS. Although comparable in diagnostic accuracy in majority of indications, endosonography retains advance in evaluation of lesions within GI tract wall (including the early stages of carcinoma), detection of small biliary stones and small pancreatic tumors. Moreover, EUS is indespensable in therapeutic indications. Experiences with fine needle aspiration, punctions and drainage of cysts and abscesses, celiac plexus neurolysis and creation of digestive anastomosis confirm its place and value among the minimally invasive procedures which minimize the need for surgical intervention, patients' trauma and treatment costs. PMID- 17087138 TI - [90Y-ibritumomab tiuxetan in patients with follicular lymphoma relapsing or refractory to rituximab]. AB - Radioimmunotherapy is a new antineoplastic treatment modality combining the effects of irradiation and monoclonal antibodies. 90Y-ibritumomab tiuxetan is a monoclonal antibody directed against the CD20 antigen to which a radioactive isotope of yttrium is attached, used for treating follicular lymphomas (FL). Using this compound we treated 8 patients with FL relapsing or refractory to combinations of rituximab and chemotherapy. Severe neutropenia developed in 5, and severe thrombocytopenia and anemia in 3 patients. Serious infections developed in 2 patients, one died. Six patients responded to treatment, 4 are still in remission after a median follow-up of 15 months, 2 died of lymphoma. Best response was achieved in low-risk patients with a low tumor burden. 90Y ibritumomab tiuxetan is an effective treatment for FL with significant hematological toxicity and a high price. PMID- 17087139 TI - [Cardiovascular diseases after kidney transplantation]. AB - Cardiovascular complications represent the leading cause of mortality in renal transplant recipients, with ischemic heart disease accounting for more than 50% of deaths. Besides the well known risk factors that affect the general population, risk for development of atherosclerosis in renal transplant patients is further increased by previous uremia and dialysis, as well as by the use of immunosuppressive agents. Diabetes mellitus, arterial hypertension, dyslipidemia, smoking, hyperhomocysteinemia, hyperuricemia, coagulation abnormalities, increased expression of cell adhesion molecules, persistent inflammation, frequent infections and obesity all increase the risk for development of atherosclerosis in transplanted patients. There is a growing body of evidence suggesting that the risk of cardiovascular disease falls significantly with smoking cessation, reduction of alcohol consumption, reduction of excessive weight, and appropriate and aggressive control of blood pressure and dyslipidemia. Patients should be instructed, and every effort should be invested to increase their compliance with the modified lifestyle and drug adherence. Novel immunosuppressive regimens tend to decrease the risk of atherosclerosis by being individualized according to the characteristics of the particular patient. PMID- 17087140 TI - [Alpha-adrenergic blockers as a support in the treatment of acute urinary retention]. AB - Catheterization remains the standard management of acute urinary retention (AUR), followed by a trial without catheter (TWOC) or prostatectomy in men who do not void spontaneously. If AUR is caused by increased sympathetic activity at the level of the prostatic smooth muscles, alpha-blockers (alpha-1 adrenoreceptor antagonists) should increase the likelihood of a successful trial without catheter (TWOC) following AUR. Alpha-blockers effectively reduce the symptoms associated with benign prostatic hyperplasia (BPH) and improve the urodynamic parameters of obstruction. They may diminish the incidence of AUR and the need for prostatectomy in symptomatic men. The adventage of tamsulosin and slow release alfuzosin over doxazosin and terazosin in the management of AUR is that a therapeutic dose can be administered at the onset of AUR, thereby reducing the time for attempting catheter removal. PMID- 17087141 TI - [Human or animal skeletal remains?]. AB - The aim of the article is to provide physicians with guidelines for differentiating human from animal skeletal remains. Morphologic features which clearly distinguish human and nonhuman bones are presented, together with an overview of the methods available for further analysis of bone material. Process of bone identification is exemplified through the case of skeletal remains recovered in Lika district in Croatia. Instructions for the physicians faced with bone material are given in order to better understand their possibilities and limitations when interpreting such findings. PMID- 17087142 TI - [Zagreb "Foundation Hospital" on Ban Jelacic Square 1804-1931 (on the 75th anniversary of its demolishion)]. AB - "Brothers of Charity Hospital" or "Foundation Hospital" in Zagreb was the oldest hospital in continental part of Croatia. It was built in 1804 on Harmica Square (present Ban Jelacic Square). The hospital building was demolished in 1931, and relocated to Sv. Duh Street where it is still located. On the 75th anniversary of the old hospital demolishion the author is showing its history (from 1804 to 1931) referring both to the particular departmens and wards development as well as to the important physicians, many of whom were the progenitors of many specialized medic professions in Croatia. PMID- 17087143 TI - [Use of erythropoietin in patients with acute renal failure]. PMID- 17087144 TI - [HT alternatives in postmenopause]. PMID- 17087145 TI - [Structural-functional organization of chloroplasts in leaves of xantha-702 mutant of Gossypium hirsutum L]. AB - For cotton mutant xantha (Gossypium hirsutum L.), it has been established that synthesis of 5-aminolevulinic acid was blocked in the light. In the light this mutant accumulates chlorophyll by 30 times lower as compared to the parent type. In mutant xantha, a very few pigment-protein complexes of PS-I and PS-II are formed in chloroplasts, and formation of membrane system in these is blocked at the early stages, in most cases, at the stage of bubbles and single short thylakoids. Functional activity of reaction centers of PS-I and PS-II is close to zero. Only light-harvesting chlorophyll-a/b protein complexes of the two photosystems are formed in mutant xantha plastid membranes with maximum chlorophyll fluorescence at 728 and 681 nm, respectively. It has been concluded that in mutant xantha genetic block of 5-aminolevulinic acid biosynthesis in the light disturbs the formation and functioning of the complexes of reaction centers of PS-I and PS-II, hindering the development of the whole membrane system in chloroplasts, causing a sharp decrease in productivity. PMID- 17087146 TI - [The influence of caldesmon on strong binding of myosin with actin in denervated rat skeletal muscles]. AB - The effect of caldesmon (CaD) on conformational changes in F-actin modified by fluorescent probe TRITC-phalloidin was investigated by polarized fluorimetry. Changes were induced by a subfragment-1 (S-1) of myosin in the absence or presence of CaD in ghost muscle fibers obtained from intact and denervated slow (SOL) and fast (EDL) skeletal muscles of rats. S-1 binding to actin of both SOL and EDL muscles was shown to cause changes in polarized parameters of TRITC phalloidin typical for a strong actin-myosin binding as well as of transition ofactin subunits from "off" to "on" state. CaD inhibits this significantly. Denervation atrophy inhibits the effect of S-1 as well but does not affect the capability of CaD decreasing the formation of strong binding in actomyosin complex. It is supposed that CaD "freezes" F-actin structure in "off" state. The denervation atrophy has no effect on CaD responsibility to bind thin filaments and to switch "off" actin monomers. PMID- 17087147 TI - [PI3-kinase inhibitors LY294002 and wortmannin have different effects on proliferation of murine embryonic stem cells]. AB - Murine embryonic stem (mES) cells can proliferate independently of the presence of growth factors in the medium. It is yet unknown what intrinsic activity triggers cell cycle events in mES cells. Here we investigated the contribution of the PI3-kinase cascade to autonomous proliferation of mES cell using PI3-kinase inhibitors wortmannin and LY294002. Wortmannin displays a weaker inhibitory effect on phosphorylation of PI3-kinase pathway target PKB as compared with LY294002, and does not downregulate mES cells proliferation, while LY294002 causes a strong decrease in the share of cells in S-phase and accumulation of cells in G1 phase. Both inhibitors cause significant decrease in cyclin D1 amount. The treatment with LY294002, rather than with wortmannin results in a decrease of cyclin E amount and cyclin E-assossiated kinase activity. In mES cells, inactivation of PI3-kinase-dependent pathway and G1 arrest are not accompanied by induction of p27kip 1 transcription and accumulation of this inhibitor of cyclin-cdk complexes at the protein level, implying that these events accomplished by some p27kip 1-independent mechanism. Both LY294002 and wortmannin cause apoptotic death of mES cells and downregulate the growth of population. Thus, inactivation of PI3-kinase in mES cells may lead to apoptosis rather than to cell cycle arrest. PMID- 17087148 TI - [Effect of plasma membrane ion permeability modulators on respiration and heat output of wheat roots]. AB - A study was made of changes in the rates of respiration, heat production, and membrane characteristics in cells of excised roots of wheat seedlings under the modulation of plasma membrane ion permeability by two membrane active compounds: valinomycin (20 microM (V50)) and chlorpromazine (50 microM (CP50) and 100 microM (CP100)). Both compounds increased the loss of potassium ions, which correlated with the lowering of membrane potential, rate of respiration, and heat production after a 2 h exposure. The differences in alteration of these parameters were due to specific action of either compound on the membrane and to the extent of ion homeostasis disturbance. V20 had a weak effect on the studied parameters. V50 caused an increase of the rate of respiration and heat production, which enhanced following a prolonged action (5 h) and were associated with ion homeostatis restoration. The extent of alteration of membrane characteristics (an increase of potassium loss by roots, and lowering of cell membrane potential) as well as energy expense under the action of CP50 during the first period were more pronounced than in the presence of V50. During a prolonged action of CP50, the increase of respiration intensity and heat production correlated with partial recovery of ion homeostatis in cells. Essential lowering of membrane potential and substantial loss of potassium by cells, starting from the early stages of their response reaction, were followed by inhibition of respiration rate and heat production. Alterations of the structure and functional characteristics of excised root cells indicate the intensification of the membrane-tropic effect of a prolonged action of CP100, and the lack of cell energy resources. PMID- 17087149 TI - [Regulatory mechanisms of mammalian imprinting]. AB - Epigenetic modifications, such as monoallelic DNA methylation, covalent histone modifications, nonhistone proteins, chromatin folding, heterochromatinization, spatial nucleus organization are reviewed with regard to establishment and maintenance of imprinting in mammals. Special attention is paid to repeated DNA sequences as intermediates of the above epigenetic modifications. A suggestion is put forward relative to importance of preimplantation development, in particular, to chromosome organization and segregation in the establishment of imprinting. Some futher directions of imprinting mechanisms are also discussed. PMID- 17087150 TI - [The chromosomal polymorphism and divergence of populations in Chironomus nuditarsis Str. (Diptera, Chironomidae)]. AB - The karyotype structure and chromosomal polymorphisms were investigated in 6 natural and 2 laboratory populations of Chironomus nuditarsis from Europe and Asia. The pool of rearranged polytene chromosome banding sequences of this species was determined that includes 16 inversion banding sequences and sequences with giant DNA-knobs (ndtG1k, ndtG2k). Obvious differences were demonstrated in the level of chromosomal polymorphism between European and Asian (Siberian) populations: the former were highly polymorphic, while the latter were practically monomorphic. It was suggested to consider the Siberian populations as marginal one. Cytogenetic distances between populations of C. nuditarsis as well between C. nuditarsis and the related species C. plumosus were estimated. The data obtained show that chromosomal rearrangements play a very important role in cytogenetic divergence of populations. PMID- 17087152 TI - The second worst deal ever. PMID- 17087151 TI - [Substrate specifity in Amoeba proteus]. AB - Three different phosphatases ("slow", "middle" and "fast") were found in Amoeba proteus (strain B) after PAGE and a subsequent gel staining in 1-naphthyl phosphate containing incubation mixture (pH 9.0). Substrate specificity of these phosphatases was determined in supernatants of homogenates using inhibitors of phosphatase activity. All phosphatases showed a broad substrate specificity. Of 10 tested compounds, p-nitrophenyl phosphate was a preferable substrate for all 3 phosphatases. All phosphatases were able to hydrolyse bis-p-nitrophenyl phosphate and, hence, displayed phosphodiesterase activity. All phosphatases hydrolysed O phospho-L-tyrosine to a greater or lesser degree. Only little differences in substrate specificity of phosphatases were noticed: 1) "fast" and "middle" phosphatases hydrolysed naphthyl phosphates and O-phospho-L-tyrosine less efficiently than did "slow" phosphatase; 2) "fast" and "middle" phosphatases hydrolysed 2- naphthyl phosphate to a lesser degree than 1-naphthyl phosphate 3) "fast" and "middle" phosphatases hydrolysed O-phospho-L-serine and O-phospho-L threonine with lower intensity as compared with "slow" phosphatase; 4) as distinct from "middle" and "slow" phosphatases, the "fast" phosphatase hydrolysed glucose-6-phosphate very poorly. The revealed broad substrate specificity of "slow" phosphatase together with data of inhibitory analysis and results of experiments with reactivation of this phosphatase by Zn2+-ions after its inactivation by EDTA strongly suggest that only the "slow" phosphatase is a true alkaline phosphatase (EC 3.1.3.1). The alkaline phosphatase of A. proteus is secreted into culture medium where its activity is low. The enzyme displays both phosphomono- and phosphodiesterase activities, in addition to supposed protein phosphatase activity. It still remains unknown, to which particular phosphatase class the amoeban "middle" and "fast" phosphatases (pH 9.0) may be assigned. PMID- 17087153 TI - Public sector buying spree. In a spate of Sunbelt deals, four publicly owned systems have bought five hospitals to adapt to market pressures. AB - Like traders in the pits, public hospital systems have been yelling "Buy, buy" in recent months. The commodities in question: not-for-profit hospitals. With a Wall Street spirit, public hospitals claim the latest acquisitions are simply a response to market pressure. Economist Stephen Zuckerman, left, calls the trend "surprising." Major pros and cons, however, give this particular investment high stakes. PMID- 17087154 TI - McGuire's billion-dollar exit. Investors are sad to see UnitedHealth chief go. PMID- 17087155 TI - Battling the 'superbugs'. CDC guidelines conflict with existing standards. PMID- 17087156 TI - Scrutiny nixes deals. Experts see no trend in failed pacts. PMID- 17087157 TI - A new direction. Baptist exec starting rural for-profit. PMID- 17087158 TI - Your opinion here. Members of healthcare community have new ways to share ideas. PMID- 17087159 TI - What patient safety looks like. Six steps that mark an organization that really cares about medical errors. PMID- 17087160 TI - So long, hospitals. Senior execs--disillusioned with the hospital industry--are trading their jobs for medical group leadership posts. PMID- 17087161 TI - In search of respect. Levine speaks from experience when she advocates on behalf of family caregivers. PMID- 17087162 TI - Shaking things up. MGMA award winner isn't afraid to tackle problems. PMID- 17087163 TI - Five years of learning from analysis of clinical occurrences in pediatric care using the London Protocol. AB - A Protocol for the Investigation of Clinical Incidents (1999) was piloted on a Winnipeg high-risk neonatal service in 2001, and was subsequently adopted as the investigative tool of choice at the Winnipeg Regional Health Authority (WRHA). The paper describes the pilot and subsequent experience with the updated London Protocol (2004) in the WRHA Child Health Program. Themes include: tightly coupled systems; multiplicity of contributory factors; medication safety; predominance of "near misses"; authority gradient; professional accountability; partnerships; and implementation challenges. The London Protocol is an invaluable tool for review of critical occurrences and near misses. To maximize impact on patient safety, healthcare organizations must involve partners and develop expertise in human factors and change management. PMID- 17087164 TI - Implementing a Good Catch program in an integrated health system. AB - In 2004, the Canadian Adverse Events Study (Baker et al. 2004) determined the incidence rate of adverse events (AE) in Canada to be 7.5%. This translates to approximately 185,000 for the almost 2.5 million annual hospital admissions in Canada. The study noted "close to 70,000 of these AEs were potentially preventable". In March 2005, a "Good Catch" program was implemented in Edmonton's Capital Health Region, one of the largest integrated health regions in Canada, as part of the region's comprehensive system of reporting, analyzing and managing incidents, adverse events and near misses. PMID- 17087165 TI - The McGill University Health Centre Policy on sentinel events: using a standardized framework to manage sentinel events, facilitate learning and improve patient safety. AB - Promoting a culture of safety within organizations includes translating the lessons learned from sentinel events into concrete changes that will improve patient safety. In May 2005 the McGill University Health Centre Policy on Sentinel Events was implemented to provide a standardized framework to manage these events and promote that culture of safety. This framework helped implement a number of changes to improve patient safety. The O2 Ticket to Ride project ensures cross-disciplinary responsibility for the transportation of oxygen dependent patients to diagnostic testing areas. The Code Stroke Algorithm was developed to expedite the sequence of events from the time the stroke symptoms are observed to the time the CT scan is carried out. PMID- 17087166 TI - Implementing system safeguards to prevent error-induced injury with opioids (narcotics): an ISMP Canada collaborative. AB - Institute for Safe Medication Practices Canada (ISMP Canada) is involved in collaborative initiatives focusing on opioid safety in two Canadian provinces: Ontario and Alberta. Baseline survey responses from these provinces indicate opportunities for improvements to the opioid system that might be applicable nationally. Information about the Ontario project and preliminary analysis of follow-up survey results from that province are shared here, to increase awareness and create further national impetus for the enhancement of safeguards in the use and management of opioids. PMID- 17087167 TI - Taking aim at fall injury adverse events: best practices and organizational change. AB - Fall injuries represent a huge healthcare, social and financial burden to the Canadian population. In 2004, the McGill University Health Centre (MUHC) was awarded recognition as a National Spotlight Organization for Implementation of the Registered Nurses Association of Ontario Best Practice Guidelines (BPGs). That same year, the author and co-leader of the Best Practice Guideline Program began the CHSRF Executive Training in Research Application (EXTRA) Program with the goal of reducing falls injuries, one of the most common adverse events in the MUHC and in acute care in Canada. This demonstration project used multiple strategies to strengthen a culture of safety and improve performance relating to adverse events, including: pilot testing several evidence-based falls prevention interventions (autumn 2005), training teams of champions to work across multiple sites, developing an infrastructure to support organizational change, modifying existing quality indicators to become benchmarkable, conducting a cost analysis of falls prevention, evaluating pre- and post-pilot surveys of organizational climate and obtaining initial baseline measures of the safety climate within the organization. Positive patient, practitioner and organizational outcomes suggest that falls safety prevention is feasible in large, complex healthcare organizations--and that safety is both a moral and a financial imperative. Next stages of the BPG program include full rollout, and measuring sustainability via a formal outcome evaluation study. PMID- 17087168 TI - Enhancing patient safety through the management of Clostridium difficile at Toronto East General Hospital. AB - In 2005 Toronto East General Hospital experienced a steady increase in the number of C. difficile cases diagnosed within the hospital. This was identified as a patient safety issue, and several areas of the hospital came together to address the problem. Pharmacy immediately started a medication review of past cases. Environmental services took the lead on the environmental cleaning, and a process was put into place with Infection Control so that housekeeping knew of every room that contained a patient with C. difficile and enhanced cleaning could be practised. Staff, including nursing, housekeeping and porters, were educated on C. difficile and the methods of transmission. A business case was developed for a disposable bedpan system, and this was approved by the senior team. A new washable product was tried out with success for the overhead patient light pulls and bathroom call bell systems. Infection rates were shared with staff through a variety of venues. As a result of the initiatives, the hospital has seen a decrease of 50% in the rates of C. difficile. A bonus was that our MRSA rates dropped as well. PMID- 17087169 TI - A multidisciplinary approach to reducing outbreaks and nosocomial MRSA in a university-affiliated hospital. AB - Endemic MRSA (methicillin-resistant Staphylococcus aureus) colonization and infection has been shown to increase morbidity, length of stay and hospital cost. Prevention of transmission demands innovative approaches. Descriptive statistics were used to determine high-incidence units. On admission, patients with a history of previous admission to a healthcare institution within the past six months were screened for MRSA. Point prevalence studies were carried out on units with more than two nosocomial (hospital-acquired) MRSA patient isolates within a four-week period. A multidisciplinary team from Infection Control and clinical units determined potential contributing factors. Recommendations included increased organism-specific education for staff, environmental cleaning and elimination of sources of transmission. Control charts to monitor nosocomial incidence rates were provided to those units that historically had a high prevalence of MRSA infections and colonization. Compliance with the infection control isolation guidelines and screening guidelines was monitored by the service. There was a 60% decrease in nosocomial MRSA between 2000 and 2001. Unit feedback was extended throughout the hospital. This decrease has been sustained since 2001 with annual rates per 1000 patient-days of 0.61 for 2000, 0.21 for 2001, 0.24 for 2002, 0.25 for 2003, 0.35 for 2004 and 0.19 for 2005. PMID- 17087170 TI - Reporting for learning and improvement: the Manitoba and Saskatchewan experience. AB - Both Saskatchewan and Manitoba have embarked on major provincial quality improvement endeavours that include a mandatory reporting and learning process aimed at enhancing patient safety by reducing the potential for recurrence of critical incidents. This move from a voluntary, less comprehensive process signals a commitment from policy makers that substantial improvements to safety will occur only when adverse events are addressed systemically within the healthcare system. Saskatchewan took the lead with the passage of legislative requirements to report, investigate and share learnings arising from critical incidents as of September 15, 2004. Manitoba is due to implement similar requirements in 2006. The focus of legislation in both provinces is aimed at reporting for learning in order to strive for further improvements in patient safety. By empowering staff and physicians to actively participate in risk identification and mitigation, both provinces have become leaders in patient safety. Saskatchewan and Manitoba have taken an innovative and collaborative approach to strive for substantive system changes, seeking out best practices in the areas of quality and patient safety. PMID- 17087171 TI - A framework for local accountability for patient safety. AB - Despite numerous publications outlining the magnitude of patient safety issues, the literature provides limited strategies for organizations to develop comprehensive, effective patient safety programs. Hamilton Health Sciences (HHS) has created a framework to foster local accountability called Patient Safety Triads and Networks. The Networks operationalize patient safety initiatives, develop knowledge and improve patient safety culture in a collaborative interdisciplinary team model. They have proven to be an effective way to support patient safety at the local level and to integrate organizational and local work on patient safety. PMID- 17087172 TI - Human factors in action: getting "pumped" at a nursing usability laboratory. AB - We describe our experience with a Nursing Usability Laboratory, where human factors design principles were applied to common nursing procedures. Our first step was to develop a human factors usability checklist. We then used this checklist while observing 11 nurses completing two standardized tasks on a simulated patient: (1) programming an insulin infusion and (2) programming a heparin infusion. We found that a usability checklist can help to uncover systematic error-provoking conditions in nursing tasks, that immediate improvements can be made in nursing training and practice and that participant nurses found the process useful. This paper will be of interest to any hospital seeking to enhance safety by applying human factors design principles. PMID- 17087173 TI - Transfer of accountability: transforming shift handover to enhance patient safety. AB - Communication of information between healthcare providers is a fundamental component of patient care. The information shared between providers who are changing shifts, referred to as "handover," helps plan patient care, identifies safety concerns and facilitates continuity of information. Absent or inaccurate information can have deleterious effects on patient care. According to the Joint Commission on Accreditation of Healthcare Organizations (JCAHO 2003), almost 70% of all sentinel events are caused by breakdown in communication. Issues and concerns regarding the effectiveness of handover at shift change were raised by nurses throughout Hamilton Health Sciences (HHS), leading to the approval of a hospital-wide project to implement evidenced-based Transfer of Accountability (TOA) Guidelines and a bedside patient safety checklist. This article describes the development of the guidelines, the results of the pilot study and the ongoing implementation of the project. The observed impact on patient safety within HHS is presented. PMID- 17087174 TI - Is your patient ready for transport? Developing an ICU patient transport decision scorecard. AB - Transport of patients from the intensive care unit (ICU) to another area of the hospital can pose serious risks if the patient has not been assessed prior to transport. Recently, the Department of Critical Care Medicine, Calgary Health Region, experienced two adverse events during transport. A subgroup of the Department's Patient Safety and Adverse Events team developed an ICU patient transport decision scorecard. This tool was tested through Plan-Do-Study-Act cycles and further revised using human factors principles. Staff, especially novice nurses, found the tool extremely useful in determining patient preparedness for transport. PMID- 17087175 TI - The role of typography in differentiating look-alike/sound-alike drug names. AB - Until recently, when errors occurred in the course of caring for patients, blame was assigned to the healthcare professionals closest to the incident rather than examining the larger system and the actions that led up to the event. Now, the medical profession is embracing expertise and methodologies used in other fields to improve its own systems in relation to patient safety issues. This exploratory study, part of a Master's of Design thesis project, was a response to the problem of errors that occur due to confusion between look-alike/sound-alike drug names (medication names that have orthographic and/or phonetic similarities). The study attempts to provide a visual means to help differentiate problematic names using formal typographic and graphic cues. The FDA's Name Differentiation Project recommendations and other typographic alternatives were considered to address issues of attention and cognition. Eleven acute care nurses participated in testing that consisted of word-recognition tasks and questions intended to elicit opinions regarding the visual treatment of look-alike/sound-alike names in the context of a label prototype. Though limited in sample size, testing provided insight into the kinds of typographic differentiation that might be effective in a high-risk situation. PMID- 17087176 TI - Patient safety in a pediatric centre: partnering with families. AB - Patient Centred Care (PCC) is a recognized pillar of quality healthcare. According to the Institute of Medicine (Kohn et al. 2000), PCC respects and is responsive to individual patient preferences, needs and values, and ensures they guide all clinical decisions. In a pediatric setting, both the child and family's preferences and values are critical; as a result, the concept of PCC is broadened to include the entire family, and is termed Family Centred Care (FCC). True FCC requires transparent and ongoing collaboration between the child, family, and all members of the healthcare team. An institution's commitment to Family Centred Care must be explicit and permeate all aspects of healthcare provisions. At Toronto's Hospital for Sick Children (SickKids), the Families as Partners in Patient Safety Committee has proven to be a successful initiative based on Family Centred Care principles. This interdisciplinary committee includes healthcare providers, parents and representatives from our hospital's Children's Council. The mandate of the group is to: (1) identify patient safety (PS) issues, (2) make recommendations to improve PS and (3) increase awareness and promote the partnership between parents and staff in PS. Key initiatives to date include developing PS information for families, a combined hand hygiene campaign and a campaign to make the hospital 100% smoke-free. A task-oriented partnership between families and healthcare workers has proven to be a productive model for advancing pediatric patient safety. PMID- 17087177 TI - "Your Health Care--Be Involved": the evaluation of a provincial patient afety tips initiative. AB - When patients take an active role in their healthcare, the results may be better, safer care. That is the premise behind the Ontario Hospital Association's (OHA) "Your Health Care--Be Involved" campaign. Launched in September 2005 by the OHA's Patient Safety Support Service, the campaign encourages active two-way communication between patients and providers and highlights the important role of patient involvement in the form of five patient safety tips. This article discusses the development, implementation and evaluation of Ontario's first-ever patient safety tips program, and what its future might hold. PMID- 17087178 TI - Cancer Care Ontario's computerized physician order entry system: a province-wide patient safety innovation. AB - More than one-third of all women and men in Canada will develop cancer during their lifetimes. Cancer patients typically require complex chemotherapy regimens, specific to their type and stage of disease, to slow or stop cancer cells from growing, multiplying, or spreading to other parts of the body. Despite the complexity of managing medication regimens for cancer patients and the associated risks to patient safety, current medical oncology practice throughout most of Canada is still to use paper-based tools, policies and procedures. To increase patient safety by reducing prescription errors and to offer clinical decision support to medical oncologists across the province, Cancer Care Ontario (CCO) developed and implemented Canada's first, cancer-specific computerized physician order entry (CPOE) system. This e-health innovation is currently in use in 11 cancer centres, and represents the largest ambulatory oncology CPOE implementation in Canada, with a 100% implementation success rate, and greater than 90% physician adoption. This paper describes the critical success factors in the design and implementation of CCO's CPOE system, including Web-based training and ease of administration to maximize physician adoption, incorporating point-of care access to clinical practice guidelines into the tool, and the use of CPOE data to monitor and increase access to anti-cancer drugs and patient safety. PMID- 17087179 TI - Predicting changes in workflow resulting from healthcare information systems: ensuring the safety of healthcare. AB - This paper describes an approach to studying medical error and workflow that can be applied to help ensure the safety of new healthcare information systems. The approach focuses on identification of usability problems resulting from implementation of new information technology, as well as identification of problems related to changes in workflow. The paper illustrates how the approach can be applied in the simulation-based analyses of emerging healthcare information systems. The paper includes discussion of the application of an approach to identifying inadvertent changes in healthcare workflow that may result from design issues in a range of information technologies including medication order entry systems. General implications for the design, development and evaluation of safer healthcare information systems are discussed. It is argued that there is a need for thorough simulation-based testing of systems under a variety of conditions before they are released in order to ensure the safety of healthcare. PMID- 17087180 TI - Frequency and type of medication discrepancies in one tertiary care hospital. AB - BACKGROUND/OBJECTIVE: Discrepancies in records used within the medication use system have been identified as a contributing factor of medication errors. The objective of this study was to determine the frequency and type of discrepancies in the medication use system in one tertiary care hospital. METHODS: Using a sample of patients (convenience sampling technique), the physician's orders, the nursing medication administration record and the pharmacy profile were compared in an attempt to identify discrepancies among them. A discrepancy was defined as a deviation from the physician's order as written in the chart. Each discrepancy was categorized according to seven components of the medication order, its location in the medication use process and its mode of delivery. RESULTS: One thousand, four hundred twenty-four orders representing 197 patients from 13 nursing units were sampled for this study. Thirteen percent of the orders were discrepant and 61% of patients had at least one discrepancy. The most frequent types of discrepancies were drug omissions and unordered drugs. DISCUSSION: The discrepancies identified in this study suggest that either orders are not reaching pharmacy or orders are not being processed appropriately in pharmacy. The location of discrepancies also suggests that there are deficiencies in communication between healthcare professionals. PMID- 17087181 TI - Broadening the patient safety agenda to include home care services. AB - Caring for an individual in the home is inherently complex. The physical environment, family dynamics and the cognitive abilities of the client and family members are only a few of the factors to be considered in delivering services. Although targeted initiatives have been established to reduce preventable injuries and deaths in the hospital sector, there has not been a corresponding level of research or patient safety initiatives in other healthcare delivery sectors. A coordinated and collaborative approach to generate new knowledge pertaining to safety in home care in Canada has therefore been undertaken by the Canadian Patient Safety Institute (CPSI), VON Canada, and Capital Health (Edmonton). Actions included the development of a background paper (Lang and Edwards 2006) that informed an invitational roundtable discussion, where key safety issues in home care were identified and priority actions discussed. Over 40 individuals from across Canada participated, reflecting various disciplinary and organizational affiliations in the delivery of home care services. This paper describes key findings from the background paper, outcomes from the ensuing roundtable discussions and implications for practice, research and policy. PMID- 17087182 TI - Adverse events among Winnipeg Home Care clients. AB - Patient safety research has focused almost exclusively on hospitals, with few studies investigating the safety of other healthcare sectors, including home care. Before measuring patient safety in home care, this study first sought to translate hospital-focused patient safety definitions and concepts to home care. A context-appropriate approach to measuring adverse events (AEs) in home care was developed using chart reviews prompted by a mixed screening process. These methods were then applied to measure the incidence, type, severity, cause, preventability and ameliorability of AEs among Winnipeg Home Care clients. PMID- 17087183 TI - Exploration of patient safety phenomena in rehabilitation and complex continuing care. AB - Patient safety has been relatively unexplored in rehabilitation and complex continuing care (CCC) settings. From the perspectives of staff members, this qualitative study aimed to explore patient safety phenomena that exist within rehabilitation/CCC and to identify the characteristics of the current workplace culture that act as enablers of or barriers to patient safety. Sixty-six staff members in a large, multisite, academic rehabilitation/CCC health centre volunteered to participate in one of six interprofessional focus groups, designed to model patient care teams that exist within the clinical programs; one focus group was also conducted with support services staff. Thematic analysis revealed that rehabilitation/CCC settings present with distinct patient safety issues due to the unique and increasingly complex populations that are served, and the place of rehabilitation/CCC along the continuum of care. Enablers and barriers identified related to teamwork, culture, resources and organizational and individual responsibility. Results of this study have helped form the foundation for future patient safety initiatives within our settings, with clear emphasis on enhancing an open and just culture in which to discuss safety issues through development of improved leadership-staff relations, teamwork and communication and clearer processes and structures for accountability. The approach to addressing these issues must fit within our rehabilitation models of care. PMID- 17087184 TI - [Post-cholecystectomy syndrome: the modern view of the problem]. AB - The authors summarize and systematize literature data and their own observations concerning post-cholecystectomy syndrome (PCES), the reasons for and the mechanisms of its development, its clinical variants etc. The authors suggest the following PCES forms should be distinguished: functional ("egenuine") forms, which develop due to gall bladder removal and the loss of its functions, and organic ("conditional") PCES forms, which develop as a consequence ofaflawy surgery and/or preoperative complications of chronic calculous cholecystitis, which dominate in the postoperative clinical picture and are mistakenly considered cholecystectomy consequences. An original operational classification of PCES is adduced; possibilities provided by contemporary instrumental and laboratory techniques of differential diagnostics are considered; differential treatment and prophylaxis of PCES are described. PMID- 17087185 TI - [Modern principles of the diagnostics and enzyme therapy of exocrine pancreatic insufficiency]. AB - The author considers the main reasons for the development of exocrine pancreatic insufficiency, discusses the advantages and disadvantages of modern direct and indirect techniques of exocrine pancreatic insufficiency diagnostics, and states the main requirements pancreatic enzymes used as means of substitutive therapy should meet. PMID- 17087186 TI - [Mucoactive therapy and new aspects of the pathogenesis of chronic obstructive pulmonary disease]. AB - From the present-day view, the article considers the potential of mucoactive therapy, directed towards the correction of mucociliary clearance (MCC) as the leading protective mechanism of the respiratory organs. Despite the big number of therapeutic agents and methods affecting the mucous link of the mucociliary system, not many of them, according to direct non-invasive in vivo studies, have a significant effect on MCC; physiotherapeutic methods have certain advantages. This means that new approaches allowing effective correction of mucociliary insufficiency should be developed. The article demonstrates the prospects of application of some pharmacological and non-pharmacological agents and development of predictive methods based on study of molecular and cellular level of bronchopulmonary pathology pathogenesis. Chronic obstructive pulmonary disease model is used to demonstrate new aspects of the pathogenesis of chronic inflammatory process in the lungs. PMID- 17087187 TI - [Polymyalgia rheumatica and giant-cell temporal arteritis]. AB - The article is dedicated to the contemporary concept of the prevalence, etiology, and pathogenesis of polymyalgia rheumatica and Horton's disease. The author analyzes the features of the pain syndrome, arthritis, fever, and other manifestations of the disease. Taking into consideration the fact that polymyalgia rheumatica and Horton's disease are usually revealed too late, the author pays special attention to their diagnostics and differential diagnostics. An increased ESR is a feature of this disease. Polymyalgia rheumatica is treated with prednisolone. The stages of glucocorticoid therapy, the rules of adjustment and lowering prednisolone doses, combined therapy with glucocorticoids and cytostatics, the effectiveness of therapy, and the prognoses are described in detail. PMID- 17087188 TI - [Cardiovascular risk factors in the main professional groups in a machine building plant]. AB - The subjects of the study were members of the main professional groups in a machine-building plant, who had long been exposed to a complex of low-intensive harmful industrial factors, not exceeding maximum allowable concentrations and levels (vibration, noise, organic solvents, neuroemotional and physical tension). The study revealed a significantly higher incidence of arterial hypertension in the members of the main professional groups and its dependence on industrial and non-industrial risk factors. PMID- 17087189 TI - [The immunity during the pre-clinical period of type I diabetes mellitus]. AB - The purpose of the study was to investigate the condition of immunity (blood lymphocyte immune phenotype and ultrastructure) in healthy children with a family background of type 1 diabetes mellitus (DM 1) having or not having diabetes associated autoantibodies (DAAB). The subjects of the study were divided into three groups. Group 1 consisted of 90 children with a family background of DM 1 (first line relatives had DM 1), DAAB- (GADA, IA-2A, and IAA) positive or negative; group 2 consisted of 51 children with newly revealed DM 1; group 3 included 45 healthy controls, normoglycemic DAAB-negative children with no family background of DM 1. GADA, IA-2A, and IAA titers were measured using radioimmunoassay. The immune phenotype of lymphocytes (CD3+, CD4+, CDr8, CD20+, and CD56+ cells) were studied using flow cytometry (FACS-analysis); their ultrastructure was studied by means of electron microscopy. The study found a significantly lower total number of T-lymphocytes (CD3+ cells), T helpers/inductors (CD4+ cells), and natural killer cells (CD56+ cells and large granule-containing lymphocytes) in the DAAB-positive children vs. the DAAB negative ones and especially the controls. In the DAAB-positive children, electron microscopy found distinct changes in the ultrastructure of CD4+ lymphocytes and large granule-containing lymphocytes (CD56+ cells), which evidences changes in the secretory and cytostatic function. Such changes in the number and ultrastructure of these lymphocyte subpopulations are found in patients with newly revealed DM 1. Thus, immune changes happen in the organism of a healthy person a long time before clinical manifestations of DM 1 develop; these changes reflect a concealed autoimmune process in Langerhans islets. Detection of DAAB plays a significant role not only in studying poorly understood pre-diabetes nature, but also in the development of new, scientifically based methods of its prevention and treatment. PMID- 17087190 TI - [Oxidative stress, the functional activity of beta-cells, and the content of tumor necrosis factor alpha in patients with type II diabetes mellitus]. AB - The purpose of the study was to investigate the effects of dietotherapy on oxidative stress (OS) condition, the fl-cell functional activity (BCFA), insulin resistance index (IRI), and the serum tumor necrosis factor alpha (alpha-TNF) level in patients with type 2 diabetes mellitus (DM 2). The subjects, 30 patients with DM 2 (9 men, 21 women), aged 42 to 70 (mean age 58.77 +/- 8.86 years), were examined. The duration of DM 2 in the subjects was from 1 month to 5 years. OS parameters, IRI and BCFA, as well as serum alpha-TNF were measured before the study and after 3 months of observation. The tests performed after the end of the study showed that hydrocarbonate exchange remained compensated, and IRI and BCFA were moderately lowered in DM 2 patients on dietotherapy. There was an insignificant elevation of serum alpha-TNF, while the condition of hydrocarbonate exchange had bettered. Changes in OS parameters in patients on dietotherapy evidence that the reserve activity of anti-oxidative system enzymes decreases. Thus, the compensation of hydrocarbonate exchange in DM 2 patients on dietotherapy retains, which is accompanied by a decrease in IRI and BCFA, while serum alpha-TNFincreases insignificantly, and reserved anti-oxidative system enzyme activity decreases moderately. PMID- 17087191 TI - [Growth factors and the hystologic picture of the liver in chronic viral hepatitis and hepatic cirrhosis]. AB - The authors of the article present the results of studying correlations between growth factors and histological changes in the liver in 48 patients with chronic viral hepatitis and hepatic cirrhosis. The results show that the blood level of transforming growth factor-beta1 (TGF-beta1) increases, whilst the level of insulin-like growth factor-1 (IGF-1), epidermal growth factors (EGF), and regeneration index (RI) in general decrease according to increase in histological activity and the degree of hepatic fibrosis. There was a positive correlation between TGF-beta1 and a negative one between IGF-1 and EGF, as well as RI, on the one part, and the degree of hepatic inflammation and fibrosis, on the other. Measurement of the serum levels of growth factors with RI calculation broadens the possibility to evaluate inflammation and fibrosis in chronic viral hepatic diseases. PMID- 17087192 TI - [The diagnostic significance of laser 13C-urea breath test in various Helicobacter pylori-associated diseases]. AB - The authors present the results of laser-based 13C-urea breath test (13C-UBT) application in diagnostics of Helicobacter pylori-associated diseases (chronic gastritis, duodenitis, and peptic ulcer). 13C-UBT data distribution before and after eradication therapy, as well as age distribution, were obtained. Correlations between the nosological forms and the rate of contamination were established. PMID- 17087193 TI - [The combination of carbohydrate and porphyrinic exchange disturbances: is it a chance or regularity?]. AB - The subjects of the study were 399 patients with internal diseases and metabolic disturbances. Carbohydrate exchange parameters (fasting level of capillary blood glucose and glucose tolerance test), and porphyrin fractions in urine (uroporphyrin, coproporphyrin), and feces (protoporphyrin, coproporphyrin) were measured. Hepatic type of porphyrinic dysmetabolism was registered in 201 (50.4%) patients. Out of these patients, 38 had disturbances corresponding to the criteria of symptomatic elevation of fecal porphyrin level, 28 had secondary coproporphyrinuria, 40 had latent, and 95 had manifest late cutaneous porphyria. In patients with normal porphyrinic exchange, the frequency of carbohydrate exchange disturbances did not exceed 6%, while in patients with different variants of porphyrinic dysmetabolism it was almost 40%. The results show that patients with hepatic type of porphyrinic dysmetabolism should be considered to have a higher risk of the development of diabetes mellitus and other carbohydrate disorders. PMID- 17087194 TI - [Disturbances of the incretory and excretory pancreatic functions in patients suffering from psoriasis in combination with chronic opisthorchosis]. AB - The aim of the study was to evaluate the incretory and excretory functions of the pancreas in patients suffering from psoriasis in combination with chronic opisthorchosis (CO). The subjects were 30 patients with psoriasis and CO, 20 patients with psoriasis without helminthiasis, 20 patients with CO, and 12 healthy individuals. The incretory pancreatic function was studied by measuring serum levels of pancreatic hormones (insulin, glucagon, and C-peptide) using radioimmunoassay with standard sets; the excretory function was studied by means of measuring serum levels of pancreatic enzymes (amylase and lipase) using a biochemical technique. Proteolytic capacity was evaluated using PABA test (per cent of the renal excretion of paraminobenzoic acid); amylolytic capacity was evaluated using coprogram results. The study revealed disturbances in the incretory and excretory functions of the pancreas in patients suffering from psoriasis or psoriasis with CO. The latter subgroup had more prominent disturbances. PMID- 17087195 TI - [The long-term results of the dynamic observation of patients with adrenal gland adenomas]. AB - The authors demonstrate the results of many years of observing patients with hormone-inactive adrenal gland adenomas. The subjects were 80 patients observed during 8 months to 12 years. The study shows that slow growth is generally typical of these adenomas. In 15% of patients an average growth of 1 mm/year was noted; this growth did not exceed 10 mm throughout the whole period of observation. The researchers also observed changes in the clinical symptoms and hormonal parameters over time. The study demonstrates that long existence of adenomas do not aggravate the clinical course of arterial hypertension and diabetes mellitus, and do not express any hormonal activity. The study established more precise indications to dynamic observation and its algorithm. PMID- 17087196 TI - [The differential diagnostic criteria and clinical features of endocrine ophtalmopathy in patients suffering from myastenia with oculomotor disturbances]. AB - The article covers clinical and ophthalmologic characteristics of oculomotor disturbances in myastenic patients with endocrine ophtalmopathy, and differential diagnostic signs and peculiarities of endocrine ophtalmopathy in patients with a combination of the two diseases. PMID- 17087197 TI - [Arterial hypertension: the modern therapeutic strategy]. AB - Arterial hypertension (AH) is a serious social and medical problem of our time. The review presents approaches to antihypertensive therapy taking into account the degree of cardiovascular complication risk; the main principles of pharmaceutical correction of AH are formulated. PMID- 17087198 TI - [Sanitary and epidemiological well-being: topical problems]. AB - The interaction of the interested services and authorities in implementing a package of goal-oriented organizational, comprehensive preventive and antiepidemic measures has given stability of the sanitary-and-epidemiological situation in the Russian Federation in some recent years. To accomplish the tasks, particularly those in developing a package of measures to reduce the prevalence of infectious diseases, the "Health" national program has been drawn up for 2006-2007 this year. One of the priorities of the program is to enhance the prophylactic orientation of the public health care system. Within the framework of this direction, it is planned to provide a further steady decline in the incidence of infections controlled by specific preventive means, primarily, in that of hepatitis B and rubella, to eliminate the congenital rubella syndrome, and to maintain the status of the Russian Federation as a poliomyelitis-free area. The subjects of the Russian Federation should redistribute allocations, by taking into account the funds envisaged in the Health program for purchase of vaccines, and direct them for implementation of other measures, such as personnel training, refrigerating equipment purchase, information-propagandistic work among the population, etc. In 2005, the poor situation established in the world due to the spread of avian influenza and there was a possible threat of influenza pandemic. Investigations ascertained the identity of influenza A virus (H5N1) isolated in the Russian Federation with the South-Eastern Asia-isolated viruses that caused mass epizooties of birds and severe diseases in persons contacting infected birds. It was assumed that 2006 spring migration might lead to the spread of pathogenic influenza H5N1 viruses throughout European Russia. This was due to the fact that the birds migrating to European Russia and a portion of birds migrating from Siberia have common winter nesting areas. To prevent mass influenza virus-infected birds' death and human infection in the epizootic localities, the federal surveillance service for protection of users' rights and human well-being jointly with the federal veterinary and phytosanitary surveillance service, and public health care administrative bodies has organized and implemented a package of measures: active headquarters have been set up in the epizooty-afflicted subjects to coordinate activities in localizing the spread of epizooty of birds; emergency commissions and antiepizootic commissions have been held; regulatory documents on the Russian Federation's subjects have been published; daily collection, analysis, systematization, and exchange of information between interested services, et. are under way. PMID- 17087199 TI - [Assessment of human health damage as a priority of human ecology and a tool for justifying managerial decisions]. AB - The paper analyzes of the currently available guidelines for assessing damages (in kind and value terms) caused by environmental factors to the population's health. The contributions of various diseases, the quality of the environment, bad habits, and other internal and external factors to the mortality structure in Russia are compared. The authors show it important to study the comparative role of the quality of life and its style, environmental pollution, the population's subjective perception of risks in the formation of health damages. PMID- 17087200 TI - [Occupational assessment and management are the basis of prophylaxis in occupational medicine]. PMID- 17087201 TI - [Cyto- and histological criteria for a risk of environment-induced diseases]. AB - The author proposes morphological criteria determining the risk of environment induced diseases. The experimental studies analyzing 23 indices of the development of reparative regeneration in the liver of rats exposed to environmental factors have revealed that a significant change in the ratio of uni and binuclear tetraploid hepatocytes is a criterion for the risk of hepatotoxic effect. Evaluation of the cytological status of nasal and oral mucosae, by using 21 indices has indicated that a significant reduction in the proportion of individuals with the normal cytological status is a criterion for the risk of environment-induced diseases. PMID- 17087202 TI - [Free radical oxidation in the assessment of health risk]. AB - Aliovalent metals (d-metals, such as manganese, nickel, iron, copper, chromium, etc.) which are able to change the degree of oxidation, to activate free radical oxidative processes, and to initiate the development of the condition that is currently defined as oxidative stress soundly and which are diagnostic criteria in the prenosological assessment of health risks are prominent among other priority ambient air pollutants. The findings suggest that there is a direct relationship between the calculated indices characterizing the carcinogenic and noncarcinogenic effects, the risk of oxidative stress, the environmental availability of prooxidants, mainly d-elements (manganese, nickel, copper, zinc) and the actual values of an increase in the rate of lipid peroxidation, of a decrease in antioxidative activity, and of an elevation in the level of methemoglobin formation. PMID- 17087203 TI - [Assessment of the sociomedical and economic damage associated with man-made ambient air pollution]. AB - The socioeconomic significance of prediction of a trend in ambient air pollution and morbidity is mainly associated with the necessity of forecasting the rate of compensation for damage to the health status of persons exposed to adverse exposures. In the industrial centers of Eastern Siberia, the health risk associated with atmospheric pollutants that have systemic toxic and irritant effects is estimated in 730 additional cases of respiratory diseases per 1,000 children a year. Medium-term forecasting of morbidity rates in the children from the town of Shelekhov, by using mathematical models, permitted estimation of society's possible health care costs. The numerical experiments made on the basis of a dynamic model has indicated that a 13% increase in the attributive risk and financial losses of dollar 3.1 million a year can be expected from the introduction of a complex of measures by the Federal special program on which about dollar 8 million are annually spent. PMID- 17087204 TI - [Approaches to assessing the human risk of mutagens]. AB - The assessment of the human risk of mutagens is a constituent of the general assessment of the risk of environment-pollutant chemicals to the population's health. An algorithm of assessing the risk of mutagens is proposed. Stage 1 (hazard identification) is to provide an expert analytical characterization of the mutagenic potential of the chemicals polluting the study object. Stage 2 (hazard characterization) is to analyze the quantitative dependences of the effect of mutagens in gametes and somatic cells of man and mammals. Stage 3 (effect evaluation) is to characterize the sources of pollution and the doses of mutagens affecting the population. Stage 4 (risk characterization) is to calculate the risk of mutagens to the population and individuals. PMID- 17087205 TI - [Analyzing the efficiency of sanitary and medical preventive measures with an allowance for criteria for human health risk]. PMID- 17087206 TI - [Estimation of environmental factors-caused damage to human health in the Republic of Kazakhstan]. AB - The author proposes a theoretical rationale for estimation of environmental factor-induced damage to human health in the Republic of Kazakhstan. The stochastic nature of damage is shown and an association of the latter with the health risk category has been established. The author has developed a model of a relationship of health life expectancy to the level of gross domestic product and a procedure for economic evaluation of environment-induced damage to human health. PMID- 17087207 TI - [Guidelines to estimating the parameters of oxidative stress upon exposure to man made environmental factors]. AB - The authors developed new methods for non-invasive diagnosis of the body's antioxidative balance, by using the kinetic characteristics of spontaneous and induced luminol-dependent chemiluminescence of an expirate. Comparative studies revealed a higher risk of diminished antioxidative protection for younger groups of the population; the total negative influence of techogenic effects and unbalanced nutrition is significant. Normalization of an antioxidative process as one of the mechanisms of formation of the body's non-specific resistance was observed in 90% of cases during medicobiological monitoring. PMID- 17087208 TI - [Indicators of the information analytical system "environmental--and hygienic risk assessment: the population's health"]. PMID- 17087209 TI - [Assessment and management of human risk due multicomponent environmental pollution in a large metallurgy center]. PMID- 17087210 TI - [Evaluation of cost-efficiency of improved life quality: methodological problems]. AB - The paper deals with theoretical approaches to and practical methods of economic interpretation and measurement of negative changes in human health, quality of life. There is evidence that it is necessary to evaluate the health status as guidelines for estimating the cost effectiveness of hygienic recommendations. The authors propose guidelines for determining the contribution of individual pollution sources. The aspects of calculation of the damage caused by water and soil pollution are considered. The lines of reforming law services to enhance the effectiveness of preventive medicine are proposed. The fact that there is a need for forming medical law is substantiated. PMID- 17087211 TI - [Development of approaches to using the parameters of oxidative balance to assess health risk due to ambient air pollutions]. PMID- 17087212 TI - [Evaluation of the sanitary-and-epidemiological safety of flocculating agents used for portable water purification]. AB - Polyelectrolytes used in the practice of water supply to the population were comparatively hygienically studied, by using a complex of hazard indices and a new approach to sanitary-and-epidemiological evaluation of the safety of water soluble polymers is substantiated. The anionic and cationic flocculating agents from different chemical classes, such as Superflok A-100, Fennopol A 321E, Fennopol K 221E, Praestol 2530 TR, VPK-402, Superflok C-577, Saipan, KF-91, Ecosol-401, a low molecular-weight sodium polyacrylate were tested as model compounds. Moreover, the information already available in the scientific literature on the toxicity of synthetic polyelectrolytes was analyzed. The generalized maximum permissible concentrations were substantiated for individual chemical classes of synthetic polyelectrolytes: polyacrylamides, polyamines, polydiallyldimethylammonium chloride. PMID- 17087213 TI - [Use of indicators for comprehensive assessment of life quality]. AB - The paper presents a procedure for quantitative sociomedical and hygienic assessment of the quality of life. Several combinations of factors that have the greatest influence on the quality of life and human health: medicodemographic indices; the prevalence and incidence rates of all classes of diseases in different age population groups; health care quality; socioeconomic conditions; natural, climatic, and sanitary conditions; quality of the population's nutrition; sociological assessment of living conditions. Based on the performed study, the indicators were estimated by each combination of factors. The quantitative determination of the integrated index of life quality may be used to assess the socioeconomic position, the health status of the population in some administrative districts and regions over time. PMID- 17087214 TI - [Environment-induced human damages caused by railway transport]. PMID- 17087215 TI - [The prevalence of respiratory diseases in the bioclimatic zones of the Primorye Territory]. AB - The paper estimates the prevalence of respiratory diseases in different bioclimatic zones of the Primorye Territory. The prevalence of environment induced respiratory diseases in the dwellers of different areas of the Primorye Teritory was found to depend on the specific features of bioclimatic zones and the combinations of environment parameters forming these zones, which should be taken into account when medical and preventive measures are implemented. PMID- 17087216 TI - [Hygienic evaluation of ambient air pollution in the industrial towns of the Ukraine and assessment of its risk to human health]. AB - The paper presents the results of a hygienic analysis of ambient air pollution in 18 large industrial towns of the Ukraine in 1990, 1996, and 2003 and calculations of cancer and non-cancer risks of atmospheric pollutions to the population's health. It also provides proposals for further development of studies. PMID- 17087217 TI - [Sociohygienic monitoring and directions in its improvement]. PMID- 17087219 TI - [Assessment of aluminum production emission induced risk to human health]. PMID- 17087218 TI - [Comparative assessment of the cancer risk of the products of smoking and ambient air pollution]. AB - Danger from aerogenic dose carcinogens entering the body with smoking products is shown to be essentially greater than that from aerogenic loading in industrial centers. The individual and population risks for smoking-induced cancer and the economic damage to the country, associated with the treatment of a large number of patients are given. To make a complete assessment of the carcinogenic risk of tobacco smoking, it is necessary to estimate all its carcinogenic constituents, their precursors, and possible transformation products at elevated temperatures. PMID- 17087220 TI - [Hygienic evaluation of the risk of man-made environmental influences on human health]. AB - In Ufa, the largest industrial center of oil-refining industry, petrochemistry, and chemistry, a long-term exposure to chemical compounds has resulted in a high chemical load on the body. A previous exposure to high doses and concentrations of a complex of pollutants caused a reduction in the total resistance of the entire population on the territory of the city. The higher mortality rates, including those due to cancer, undoubtedly reflects both the exposure to delayed effects of a complex of chemical compounds, the total level of which exceeded the hygienic standards, and the combined effects of a whole complex of pollutants over 25-30 years. Therefore, on assessing the risks of technogenic action on the health status of the population of a city with the traditionally petrochemical and chemical profile under the conditions of complex pollution, it is necessary to take into account the priority regional features: the delayed presentation of adverse environmental exposures. PMID- 17087221 TI - [The environmental-and-hygienic risk of industrial waste and consumption]. AB - How to safely handle production and consumption residues is a relatively recent problem. According to the data of the United Nations Organization, 25 to 33% of the diseases notified in the world are directly related to the poor human environment. In Russia and foreign countries, there has been no list of chemicals so far. By taking into account the general spread of human vital activity-induced production and consumption residues, the substantial scale of formation and a great variety of the latter's in their composition, types, and the pattern of their possible deleterious action, it is important to consider the problem of handling the residues, by assessing the risk of their hazard. PMID- 17087222 TI - [Rationale for the integral life quality index]. PMID- 17087223 TI - [Significance of major factors forming the demographic setting]. AB - The time course of changes in ambient air pollution was hygienically estimated, socioeconomic factors (n = 31) and level of health care delivery (20 factors) were studied in selected towns (a city numbering over million dwellers, a large industrial center, and an average numbered administrative town). The factors forming the population's health were stratified in score, by taking into account the weighting factors (Delhi's method) in relation to the values of an administrative town, which were taken as 1 (one). The authors are the first to show that in industrial centers, natural health losses are chiefly determined by the influence of polluted ambient air (3.4 and 2.3 scores versus 1.0 score in the administrative town). There were virtually no differences in socioeconomic conditions (1.1 and 1.0 score). The quality of health care provision is higher in the million-numbered city (1.6 scores) and in large town (1.2 scores). The specific contribution of factors to the demographic setting was defined under specific conditions. PMID- 17087224 TI - [Human risk perceived by different population groups in case of Sayanovsk dwellers]. PMID- 17087225 TI - [Characterization of quality of life in schoolchildren]. PMID- 17087226 TI - [Role of genetic factors in adolescents' dysadaptive behavior]. PMID- 17087227 TI - [Use of laser correlation spectroscopy for evaluation of metabolic changes in workers engaged in radiation-dangerous industry]. AB - The pattern of metabolic changes was studied in nuclear fuel plant workers by laser correlation spectroscopy (LCS) of biological fluids (blood serum and plasma, urine, oropharyngeal lavages (OPL). Plasma samples were divided into 3 groups: 1) control (unirradiated) samples; 2) those irradiated by below 100 mZv; 2) those irradiated by more than 100 mZv. With larger dose irradiation, the contribution of small particles (6-8 nm) to the dispersion of increased and the proportion of large components (300-400 nm) decreased. There was a correlation between the total accumulated dose, the dose in the past 9 months and the changes in the contribution of the above groups of particles to light diffusion. The found regulations in the changes of the subfraction composition of blood agree with earlier data on changes in the serum of nuclear fuel workers. Analysis of urine samples revealed an increase in the contribution of catabolic processes. That of OPL showed the preponderance of anabolic changes over catabolic ones in the presence of a considerable contribution of normologically similar LC spectra. Differences were found in the pattern of metabolic changes in relation to technological stages. Although the nature of the observed spectral transformations remains unknown, the simplicity and rapidity of the LCS technique may be considered as a suitable tool for detecting the effects caused by small dose irradiation and other factors. PMID- 17087228 TI - [Assessment of risk to the Minsk population's health due to ambient air-polluting chemicals]. PMID- 17087229 TI - [Criterion evaluation of oxidative-and-antioxidative processes in the body's biological media in the hygienic diagnostic chemical factors]. PMID- 17087230 TI - [Improvement of methods for assessing public health; use of the parameters of standardized and cumulative risks of death]. AB - The paper gives the results of an analysis of interval estimations of the indices and age-adjusted structure of malignancy mortality, by using direct, indirect, and cumulative standardization methods. General trends in cancer mortality have been revealed and an excess risk of cancer deaths and the structural features of mortality due to cancer of the respiratory system and chest and gender-specific pathology: breast cancer in females and prostate cancer in males have been assessed in the Kiev Region in 1980-2003. PMID- 17087231 TI - [The medical and demographic situation in the country]. PMID- 17087232 TI - [Problems in the organization of prevention of environment-induced abnormalities]. PMID- 17087233 TI - [Health indices, which characterize the efficiency of the sociohygienic monitoring system, in the Moscow population]. AB - The paper presents some indices that characterize morbidity in the Moscow population, which suggest positive trends in the health status in all population groups. These include stabilization of overall morbidity rates; a down-increase in the incidence of respiratory diseases and bronchial asthma; a reduction in overall morbidity in infants of the first year of life, including a reduction in the incidence of life-threatening diseases (perinatal pathology and congenital malformations; a decrease in the incidence of alimentary diseases associated with social factors. The found positive changes reflect the efficiency of introduction of the sociohygienic monitoring (AHM) system. One of the mechanisms of introduction of AHM results is to submit data to the Moscow Government in the annual reports "On the Moscow population's health status" which cover demography, morbidity, female health, and the regional features of the population's health in relation to environmental factors. Based on information, managerial decisions and measures to improve the sanitary-and-epidemiological situation in Moscow are taken and implemented. PMID- 17087234 TI - [Classification of hazards of industrial enterprises on a new methodological basis]. AB - Based on the analysis of data on the volume of emission from 163 enterprises, a classification of industrial projects that are the sources of ambient air pollution has been developed, which includes: an estimate of the degree of hazard (5 classes) from enterprises by the total volume of emission irrespective industrial affiliation and the kind of activities; the relative index (RI) of hazard from an enterprise, which considers the volume of emission and the degree of toxicity of each pollutant (the RI specifies the class of an enterprise); the possible sizes of sanitary-hygiene zones in relation to the total volume of emission (tons per year). Testing the developed classification, by using 64 enterprises and various projects as an example has shown the expedisanitary hygiene zones of an enterprise and projects of any branches of industry or the kind of activities, which are ambient air pollution sources. PMID- 17087235 TI - [Use of health risk estimates for substantiating the boundaries of sanitary protection zones of the Novolipetsk metallurgic works]. PMID- 17087236 TI - [The country's geochemical provinces and the population's health]. AB - The paper describes possible conditions for forming the natural and artificial geochemical provinces available on the territory of the Russian Federation, as well as the impact of localities' soil pollution with chemicals on human health, by using specific areas as an example. Emphasis is laid on the fact that it is urgent to substantiate priorities to organize and manage a section of sociohygienic monitoring of localities' soil. PMID- 17087237 TI - [Hygienic standards for ambient air pollutants: the-state-of-the-art and ways of harmonization]. AB - The paper presents data on the harmonization of hygienic standards for ambient air pollutants and perspective lines for further development of these studies. PMID- 17087238 TI - [NO-synthase activity products and respiratory tract inflammation: metabolism, pathophysiological role in allergic diseases]. PMID- 17087239 TI - [Association of the levels of brain and atrial natriuretic peptides with the concentrations of inflammatory markers in patients with congestive heart failure]. PMID- 17087240 TI - [Once more about the determination of total protein in urine]. PMID- 17087241 TI - [Hair trace elements in patients with goiter]. AB - The investigation determined whether there was a relationship of thyroid diseases with the geology of the place of residence and its associated spectrum of trace elements (iron, chromium, selenium, manganese, cobalt, copper, nickel, arsenic, and zinc). The geologo-geomorphological factors of the habitat, such as geological formations (mineral composition, structure, the degree of rock metamorphism), tectonics, the history of geological development, relief, paleorelief, were found to be of considerable importance in the formation of the human trace element status. Both deficiency and excess of trace elements was shown to be of pathogenetic value in the development of thyroid diseases. Residence in the area wherein carbonate rocks are predominant contributes to an increase in incidence of thyroid diseases. PMID- 17087242 TI - [Neutrophilic nuclear segmentation: a new view of the nature of the phenomenon]. PMID- 17087243 TI - [Lipopolysaccharide-induced apoptosis of peripheral granulocytes in patients with prior salmonella infection]. AB - The in vitro induction of granulocytes in the presence of high and low concentrations of lipopolysaccharides (LPS) allowed the authors to study a dose dependent apoptotic response in the population of intact cells and in that of primed cells in the period of convalescence in patients with Salmonella infection. An experimental system was used to study a respiratory burst and granulocytic apoptosis in the presence of hypothalamic neuropeptides. The clinical manifestations of the diseases were studied in 32 inpatients. It was shown that there was an increase in the sensitivity of granulocytes to the apoptotic action of low, suboptimal doses (100 ng) of S. enteritidis LPS in patients with salmonellosis at convalescence. This correlates with the severity of a clinical course of the experienced disease and with the suppressed sensitivity of apoptosis to the optimal doses (1000 ng) LPS. PMID- 17087244 TI - [Standardization of coagulologic technique for antithrombin III estimation and its clinical application]. AB - A standard assay procedure for estimating antithrombin III activity was derived from the Abildgaard technique as follows: 1) to decrease errors at the preanalytical stage, by using preliminarily defibrinated lyophilized plasma calibrator and 2) to increase measurement accuracy, by using the concentration of a fibrinogen preparation and simplifying the analytical procedure. The estimation of plasma antithrombin activity in patients with pulmonary thromboembolism or sepsis points to the changes in antithrombin III activity. PMID- 17087245 TI - [Low molecular-weight heparin (clexane) in the prevention of thrombotic events in patients with malignancies]. AB - The basic links of the hemostatic system and the markers of intravascular thrombogenesis were studied, by using an automatic STA Compact analyzer in 80 patients with locomotor tumors. The patients were divided into 2 groups: 1) 50 patients received clexane, 40 mg, 12 hours before and within 7-18 days after surgery (a study group); 2) 30 patients underwent only nonspecific prevention of thrombotic events (a control group). Postoperatively, hypercoagulation was shown to persist long (up to 18 days) in patients with locomotor tumors. Clexane used in the pre- and postoperative periods within 18-20 days was effective in reducing the rate of intravascular blood coagulation, preventing thrombotic events, and failing to cause hemorrhages. PMID- 17087246 TI - [Biological properties of hospital-acquired Klebsiella pneumoniae strains]. PMID- 17087247 TI - [Detection of Borrelia DNA in the Borrelia burgdorferi sensu lato complex in the blood of patients with Ixodes tick-borne borrelios]. AB - Borrelia DNA was detected by polymerase chain reaction in the blood of patients who had suffered from tick suction. DNA was revealed in 28.7 of the blood samples from patients with the erythematous form of Ixodes tick-born borrelioses (ITBB) and in 14.3% of those diagnosed as having tick-borne encephalitis. Blood Borrelia DNA was not detected in patients with end-stage and chronic ITBB. Comparing the results of detection of DNA with those Borrelia protein antibodies has shown that the antibody titers detectable in the patients having Borrelia DNA are lower than those in the patients with the same form of ITBB and without DNA. The detection of DNA should be accomplished in the first 4 weeks after tick bite. PMID- 17087249 TI - [Laboratory information systems: use, possibilities, and recommendations]. PMID- 17087248 TI - [To assay of the concentration of Ureaplasma cultures in the liquid nutrient medium]. PMID- 17087250 TI - [The goals and objectives of the Association of Clinical Laboratory Diagnostic Means Manufacture]. PMID- 17087251 TI - [Domestic manufacture of means for clinical laboratory diagnosis]. PMID- 17087253 TI - Therapy for the soul. PMID- 17087254 TI - Equipped for fitness. PMID- 17087255 TI - Mobilizing youngsters. PMID- 17087257 TI - Industry insiders offer paper-free connection. PMID- 17087256 TI - The paperless trail. PMID- 17087258 TI - When it's more than a scratch. PMID- 17087259 TI - Going back to basics. PMID- 17087260 TI - The latest and the greatest. PMID- 17087261 TI - Protecting weekend warriors. PMID- 17087262 TI - Definitive new guidelines. PMID- 17087263 TI - [Diagnosis and therapy for pituitary tumors]. PMID- 17087264 TI - [Transnasal tumor exploration in infradiaphragmatic craniopharyngiomas]. PMID- 17087265 TI - [A novel method of dural repair using polyglycolic acid non-woven fabric and fibrin glue: clinical results of 140 cases]. AB - This paper presents a report based on the results obtained from clinical applications of a biocompatible dural substitute made of polyglycolic acid non woven fabric and fibrin glue. The cases subjected to this study were the ones needing reconstruction of dura mater which had become defective due to injury or brain tumor and the ones in which primary suture of the dura mater was considered to be too difficult or inadequate with ordinary methods. The dural substitute was used in 140 cases during the period between June, 2001 and July, 2005. The operations were performed using the supratentorial approach in 66 cases and infratentorial approach in 74 cases. Among these procedures, 39 cases were indicated for microvascular decompression, the commonest operation performed, then cranial base surgery in 27 cases and tumor resection in 24 cases, and so on. Lumber spinal fluid drainage or re-operation was required in 3 cases (2.1%) due to formation of post-operative cerebrospinal fluid leakage or subcutaneous accumulation of cerebrospinal fluid. With the dural substitute no infection was observed as a complication in any of the cases. Among the 140 cases presented this time, 27 cases were cranial base surgery and 74 cases were performed, using the infratentorial approach. Nevertheless, the study showed that the closing ability of the dural substitute was adequate even in actual clinical settings it is reported above that the incidence rate of post-operative cerebrospinal fluid leakage or subcutaneous accumulation of cerebrospinal fluid which require additional intervention was only 2.1%. PMID- 17087266 TI - [Surgical treatment for cerebrospinal fluid rhinorrhea from plaum sphenoidale]. AB - A transcranial approach is frequently employed for the closure of cerebrospinal fluid (CSF) rhinorrhea, occurring after skullbase surgery or associated with anterior skull base fracture. However, the site of CSF rhinorrhea is usually most distant from the site of craniotomy and, because securing of an adequate operative field is difficult, elimination of this type of CSF rhinorrhea tends to be insufficient. We have attempted to reconstruct the anatomical structures with an extended transsphenoidal approach for a case with CSF rhinorrhea from the planum sphenoidale, and have succeeded in achieving a more physiological and reliable elimination of CSF rhinorrhea. This technique is presented, with a representative example. PMID- 17087267 TI - [A case of symptomatic arachnoid cyst of the occipital convexity in an adult]. AB - Symptomatic arachnoid cyst in an adult patient is rare. We present a case of a 51 year-old female with an arachnoid cyst of the right occipital convexity who developed homonymous hemianopsia. She had complained of numbness in the limbs at the age of 47 and based on MRI was diagnosed with a cystic space-occupying lesion in the right occipital convexity. After being under observation for four and a half years, she complained of headache and visual disturbance, at which time we diagnosed growth of the cyst. On ophthalmologic examination, homonymous hemianopsia was demonstrated. She underwent membranectomy, which resulted in the disappearance of visual disturbance. This case is comparatively rare among previously reported cases of symptomatic arachnoid cyst with regard to both location and symptom. Membranectomy is one of effective methods for treating symptomatic arachnoid cyst in the occipital convexity. PMID- 17087268 TI - [Two cases of main trunk artery occlusion associated with multiple cerebral aneurysms]. AB - We report 2 cases of multiple aneurysms (AN) associated with main trunk artery occlusion. CASE 1: A 52-year-old male was admitted to our hospital with dysarthria and weakness of the right side of the body. Computed tomography (CT) showed cerebral infarction in the left corona radiata. MR angiography and conventional angiography showed occlusion of the left middle cerebral artery (MCA) and saccular aneurysms (ANs) at the origin of the anterior communicating artery (A-com) and bifurcation of the right MCA. Subsequent 123I-IMP-single photon emission tomography (SPECT) revealed marked reduction of cerebral blood flow and disturbed reactivity to acetazolamide in the left cerebral hemisphere. Superficial temporal artery (STA)-MCA anastomosis was performed to improve cerebral blood flow and reduce hemodynamic stress for AN of the A-com and right MCA. At 5 months after the first operation, neck clipping was performed successfully for the non-ruptured A-com AN and right MCA AN. CASE 2: A 65-year old male was admitted to our hospital. CT revealed subarachnoid hemorrhage (SAH), and 3D-computed tomographic angiography (CTA) and cerebral angiography showed basilar top AN, A-com AN and right MCA AN associated with right internal carotid artery occlusion. Right ACA and MCA territories were visualized from the A-com artery and posterior cerebral artery. STA-MCA anastomosis was performed to improve cerebral blood flow and reduce hemodynamic stress for ANs. In the same operation, successful neck clipping was performed for BA top AN and right MCA AN. In such cases as these, particularly in ischemic cases associated with main trunk artery occlusion, it was important to consider surgery for AN after STA-MCA anastomosis in anticipation of improved cerebral blood flow and reduce hemodynamic stress for AN. PMID- 17087269 TI - [Treatment of a ruptured giant internal carotid artery pseudoaneurysm following transsphenoidal surgery: case report and literature review]. AB - We report here a case of giant internal carotid artery (ICA) pseudoaneurysm as a complication of transsphenoidal surgery. This 50-year-old acromegalic male presented to our clinic with a status of hypovolemic shock due to serious epistaxis. Neuroradiological examinations at his admission revealed a giant aneurysm in the right cavernous portion projecting into the sphenoid sinus. Eight years before this presentation, he had undergone a transsphenoidal surgery for growth hormone producing pituitary tumor at the other clinic. Then intraoperative arterial bleeding was reported, probably as a result of carotid injury. His medical history and radiological findings suggested that his epistaxis resulted from a rupture of the iatrogenic pseudoaneurysm which had gradually grown after the ICA injury at the previous surgery over 8 years. Emergent coil embolization using Guglielmi detachable coils (GDCs) resulted in a successful homeostasis. Major part of the aneurysm dome was obliterated via the intervention, however small part of the aneurysm neck was unable to be obliterated due to a technical difficulty. His postoperative course was favorable, but he suffered from a recurrence of serious epitaxis 4 weeks after the embolization. Emergent angiography suggested a rupture of the un-obliterated aneurysm neck remnant. Thus, trapping of the aneurysm combined with high flow bypass was necessitated. Relevant literatures are reviewed, and possible therapeutic strategies for this rare lesion are discussed. PMID- 17087270 TI - [A case of the myelitis due to Toxocara canis infection complicated with cervical spondylosis]. AB - A 59-year-old male had suffered from numbness of the hands for 7 months. With a diagnosis of cervical spondylosis, he had been treated conservatively at a nearby clinic. After he fell off his bicycle, the numbness intensified and limb weakness developed. Cervical MRI revealed spinal cord compression at the C4/5 and C5/6 levels due to cervical spondylosis with prominent edema in the spinal cord spreading from the C4 to C6 level. The edema was very serious. Therefore, we suspected that the traumatic spinal injury underlying the cervical spondylosis was complicated by another disease. Cervical spinal angiography revealed no apparent vascular disorder. Contrast enhanced MRI showed a small enhanced area in the spinal cord at the C5 level. Because of the rapid progression of gait disturbance, expansive laminoplasty was performed without further examination. Although remarkable amelioration of the symptoms was seen just after the surgery, the symptoms worsened again about 1 month later. The patient's clinical history was reconsidered, revealing that he likes raw bovine liver. Serological examination, because of suspicion of parasitic infection showed elevated titers of anti-Toxocara canis antibody in the serum and cerebrospinal fluid. Administration of albendazole improved the clinical symptoms, and normalized the serological and MRI findings. Myelitis due to T canis infection is a rare disease. For an early and accurate diagnosis, it is important to be fully aware of this disease and to include detailed information on food preferences and pet keeping in the process of compiling a clinical history. PMID- 17087271 TI - ["Disease targeting therapy" for neurosurgical disorders: molecular targeting and drug delivery]. AB - Recent progress in molecular biology introduced a concept of molecular targeting therapy against many diseases including neurosurgical disorders. Molecular targeting therapy is promising, because targeting a disease-specific molecule it may provide cure without inducing unfavorable side effects. Focusing mainly on CNS (central nervous system) malignancies, authors discuss the current development of molecular targeting therapy with a special concern to strategies using DNA, RNA, immunotoxin, and molecular selective inhibitors. Strategies using DNA include gene therapy and DNA vaccine. RNA interference (RNAi) is one of the promising molecular targeting strategies using RNA. Immunotoxins such as recombinant chimeric cytotoxin consisting of interleukin-13 and a truncated exotoxin produced by the Pseudomonas aeruginosa bacterium is now under clinical investigation. Many molecular targeted agents such as selective inhibitors for protein kinases are now in preclinical and clinical development for patients with malignant glioma. Additionally, authors introduce their attempts to visualize the local drug delivery using real-time MRI monitoring. Visualization of drug delivery, while achieving effective drug distribution using convection-enhanced delivery (CED), may also contribute to the disease targeting. These strategies together should help us develop the effective 'disease-targeting therapy' for CNS disorders. PMID- 17087272 TI - [Carotid and coronary artery disease associated with brain infarction]. PMID- 17087273 TI - [Imaging of cardiac sources of cerebral embolism]. PMID- 17087274 TI - [Treatment strategy for concomitant cervical carotid artery and coronary artery diseases]. PMID- 17087275 TI - [Primary and secondary prevention of cardiogenic cerebral embolism]. PMID- 17087276 TI - [Quality assurance of medical treatment and preventable trauma death]. PMID- 17087277 TI - [The questionnaire survey to clarify correspondence of medical doctors and dentists in Fukuoka City, Japan, for warfarin therapy at the dental extraction]. AB - PURPOSE: The questionnaire survey was performed in order to clarify correspondence of medical doctors and dentists working at hospital service for warfarin therapy at the dental extraction. METHODS: For 17 senior dentists in 17 hospitals and 142 senior physicians in 82 hospitals in Fukuoka City, we investigated using the questionnaire whether they supported dental extraction under warfarin therapy or not in patients having warfarin treatment with past history of cardioembolic stroke due to non-valvular atrial fibrillation (NVAF). Specialty and experience of stroke after withdrawal of warfarin were also asked in medical doctors. RESULT: The reply was obtained from 14 dentists (82.4%) and 66 doctors (46.5%). The number of dentists and doctors who pull out a tooth under warfarin treatment continuation were 12 persons (85.7%) and 32 persons (48.5%), respectively. The rates in doctors whose specialty was stroke, cardiovascular disease, and others were 78.6% (11/14), 36.0% (9/25), and 44.4% (12/27), respectively. The rate in stroke specialists was significantly higher than that in cardiovascular disease specialists (p = 0.019). The stroke specialists had experience of stroke after withdrawal of warfarin more frequently than the cardiovascular disease specialists (57.1% vs. 20.0%, p = 0.033). CONCLUSION: The rates of acceptance in tooth extraction under warfarin treatment continuation of the dentist were high in Fukuoka city, while that of the medical doctors were not. Rate of doctor's acceptance differed according to the specialties, and may be affected by experience of stroke after cessation of warfarin therapy. PMID- 17087278 TI - [A detection of very mild Alzheimer's disease and MCI by memory complaints and deterioration of IADL in a questionnaire for a memory clinic]. AB - BACKGROUND AND PURPOSE: There have been very few studies trying to explain about daily life features of patients with very mild Alzheimer disease (AD) and of those with mild cognitive impairment (MCI). The purpose of this study was 1) to clarify characteristics of very mild AD and MCI in their daily life, and 2) to examine items in a questionnaire that can be useful for detecting subjects of suspected AD. SUBJECTS AND METHODS: Subjects were 111 patients of the memory clinic in National Center Hospital for Mental, Nervous, and Muscular Disorders, National Center of Neurology and Psychiatry, Japan; 39 normal range, 28 MCI, and 44 very mild AD. On their first visits, they were asked to fill out a questionnaire consisted of 103 items. Thirty five items in a questionnaire were selected, 15 memory-complaint items and 20 items of instrumental activities of daily living (IADL), and examined retrospectively. At first we compared each set of groups using the chi-square test. Then a logistic regression analysis was used to observe which items contributed to judge a person to be suspected of AD. And further, sensitivity and specificity for discriminating between an AD suspected and a normal range were examined. RESULTS: None of the items showed statistical significance between MCI group and very mild AD group. Then, we combined these two groups into one group (n = 72), and made comparison between this new group and normal range group (n = 39). There were 18 items that showed statistical significance between the two groups. And five of them, 'dysfunction of memory: noticed by others', 'dysfunction of memory: getting worse', 'unable to utilize the memorandum', 'forget incidents occurred a few days ago' and 'unable to locate unfamiliar places' showed significantly high odds ratio in judgment on a person to be suspected of AD. In addition to preceding 5 items we adopted two items, 'dysfunction of memory: unnoticeable by her/himself' and 'unable to manage household expenses', which were not included in the logistic regression analysis, but were essential for the discrimination between the two groups. The sum of score of these 7 items showed high specificity and sensitivity for dividing between the two groups. CONCLUSION: These findings indicated that an exploration of memory complaints and deteriorations of IADL was useful for the detection of MCI and very mild AD, especially 7 items described above could be used as a simple questionnaire for the elderly. PMID- 17087279 TI - [Parkinsonism secondary to chronic subdural hematoma. A case report]. AB - We described a case of secondary parkinsonism due to chronic subdural hematoma (CSH). An 83-year-old man developed memory impairment and gait disturbance gradually since one year ago, and these symptoms subacutely deteriorated after he fell down and hit his head. On admission he had moderate cognitive impairment, gait disturbance (wide-based, small steps and initial freezing), postual instability, akinesia and right-sided rigidity. He did not have resting tremor. A CT scan showed a right-sided CSH with marked compression on the left hemisphere, and remarkable decrease of the blood flow in the left cerebral hemisphere was revealed by single photon emission computed tomography. His neurological condition improved gradually through the decrease of the CSH, which suggested the diagnosis of secondary parkinsonism due to CSH. We thought that the CSH in the right hemisphere caused right-sided parkinsonism through mechanical compression to the left hemisphere, which was shown in the SPECT image, in this case. Elderly people suffering from subacute progressive parkinsonism, should undergo CT studies be to differentiate the parkinsonism secondary to CSH. PMID- 17087280 TI - [Cerebral artery lesion in a patient with infective endocarditis: serial MRI and MRA findings of cerebral artery stenosis]. AB - We reported a case of infective endocarditis (IE) complicated with bacterial meningitis and cerebral artery stenosis. A 22-year-old man was admitted to our hospital because of IE. Although benzylpenicillin administration was continued, he abruptly developed consciousness disturbance on the seventh day. His cerebrospinal fluid indicated bacterial meningitis. MRI with gadolinium (Gd) enhancement showed septic embolism in the left parietal lobe and bi-linear enhancement on the right middle cerebral artery (MCA). MRA demonstrated narrowing of the MCA at the same site as the bi-linear Gd enhancement. We considered that these findings show narrowing of the MCA was due to cerebral arteritis. Intravenous administration of ampicillin and cefpirome gradually improved both IE and cerebral artery stenosis. We wish to emphasize that combination of MRI with Gd enhancement and MRA may be useful not only for diagnosis of cerebral artery stenosis but also for evaluation of treatment effect. PMID- 17087281 TI - [A case of pure agraphia due to left parietal lobe infarction]. AB - We reported a case of an 86-year old woman with pure agraphia due to the left parietal lobe infarction. The characteristics of agraphia were as follows. Most errors in Kana and Kanji writing to dictation and copying were no response. She was able to write only numerals from 1 to 12 precisely. Most errors in numerals were substitution. One unrecognizable numeral was found. She succeeded in pointing to nine among ten single Kana letter named by the examiner with the systematic table of the Japanese syllabary, but missed in pointing to Kana words. It took more time for the patient to point to single Kana letter than the control. Magnetic resonance imaging showed a cerebral infarction in the left parietal lobe which included a part of superior parietal lobule and supramarginal gyrus. We classified pure agraphia with parietal lobe infarction into two types in our previous report. In one type (type 1), letters in writing are poorly formed, but the ability to make words with the methods other than writing are reserved. The only deficit of graphic motor pattern could cause Type 1 agraphia. In another type (type 2), letters in writing were well-formed, but spelling with anagram or typing was disturbed. The deficits of writing process other than graphic motor pattern could cause Type 2 agraphia. This typing seems to be effective not only in Kana but also in Kanji. In this report, we investigated the differences of lesion between two types out of some references. Type1 agraphia seems related to lesion of left superior parietal lobule, while Type 2 agraphia seems related to lesion of left supramarginal gyrus. This case had the features of type 2 agraphia at least, and the compatible lesions. PMID- 17087282 TI - [A case of idiopathic thoracic spinal cord herniation]. AB - We report a case of idiopathic thoracic spinal cord herniation. A 63-year-old female came to our hospital complaining of a slowly progressive gait disturbance that began 5 years ago. She showed signs of a left dominant spastic paraparesis and sensory disturbance. Her left foot had already became a drop foot from 2 years ago. Magnetic resonance (MR) imaging and computed tomographic (CT) myelography demonstrated that the atrophic spinal cord had displaced to the ventral side with a dilated dorsal subarachnoid space at T2-3 level. Surgery was performed via T2-3 laminoplastic laminectomy. The ventral aspect of the spinal cord revealed a defect in the inner layer of the duplicated dura mater, into which the gliotic spinal cord herniated. The herniated spinal cord was repositioned and the dural defect was repaired using a GORE-TEX dura substitute. Postoperative course was uneventful. Although the patient showed marked improvement in the right lower limb, there was slight improvement in the left lower limb. The clinical symptoms of this disease are uncommon and progress slowly, and if left untreated will result in paraparesis. Therefore early diagnosis and operation is important to prevent the progression of symptoms and to achieve a satisfactory outcome. PMID- 17087283 TI - [A case of Tolosa-Hunt syndrome demonstrated by fat-suppression MR images]. PMID- 17087286 TI - [Endothelial dysfunction and plaque inflammation in diabetic macroangiopathy]. PMID- 17087287 TI - [Atherosclerosis in diabetes mellitus]. AB - The clinical and epidemiological importance of cardiovascular diseases, has increased rapidly in recent years. Diabetes mellitus which serves as a basis for the development of atherosclerosis, are induced by a genetic predisposition coupled with environmental factors. Hypernutrition and insufficient exercise lead to diabetes mellitus, hyperlipidemia, hypertension, obesity, and insulin resistance in individuals genetically predisposed to these disorders, which eventually produce ischemic heart disease and cerebrovascular disease. The report of the Multiple Risk Factor Intervention Trial (MRFIT) suggested that the prevalence of atherosclerosis in patients with diabetes mellitus is two or three times that in individuals without diabetes, and a Finnish study has reported that the risk of ischemic heart disease is extremely high in patients with type 2 diabetes mellitus. In order to prevent the development of ischemic heart disease in patients with diabetes mellitus, the risks of development of hyperlipidemia, hypertension and other relevant diseases should be carefully controlled. PMID- 17087288 TI - [Mechanism of diabetic macroangiopathy]. AB - Diabetes is a strong risk factor for cardiovascular diseases. Hyperglycemia at post-prandial as well as fasting state explains a part of the risk. Several disorders induced by hyperglycemia such as activation of PKC, increased glycosylation and oxidative stress have been proposed as the underlying mechanisms. In addition to the hyperglycemia, several epidemiological studies have suggested that hyperinsulinemia or insulin-resistance is also associated with the cardiovascular diseases. Endothelial dysfunction and/or inflammation are postulated as key players in the hyperinsulinemic or insulin-resistant state. Recently, we found that insulin stimulates the proinflammatory gene expression in cultured vascular cells. In this article, I reviewed these hypothesis explaining how hyperglycemia or insulin-resistance caused macroangiopathy in diabetics. PMID- 17087289 TI - [Glucose intolerance: postprandial hyper glucose, risk factor for diabetic macroangiopathy]. AB - Major prospective studies about diabetes showed simply improving glucose control is not sufficient to prevent diabetes macroangiopathy. Hence, postprandial hyperglycemia is focused as new risk factor of diabetes macroangiopathy. It is getting common that IGT (impaired glucose tolerance) is a major risk of cardiovascular disease. Our previous data suggest that the risk of macroangiopathy is not postprandial hyperglycemia itself but pathophysiology of causing postprandial hyperglycemia; reducing insulin sensitivity and/or delayed and hyper insulin level. To prevent postprandial hyperglycemia, there are three strategies according to pathophysiology. Improving insulin sensitivity of liver, delaying glucose flux to portal vein and reviving fast insulin secretion. Recent study shows each strategy can reduce risk of diabetes macroangiopathy. PMID- 17087290 TI - [Pathological characteristics of diabetic macro-angiopathy]. AB - The pathological features of coronary diabetic macro-angiopathy were characterized by the acceleration of atherosclerotic lesions. Among the Hisayama cohort study subjected for participants in the screening examination in 1988, the coronary atherosclerotic grade, pathologically estimated on luminal stenosis and/or incidence of advanced lesions of coronary arteries, was significantly correlated with serum HbA1c, levels. These findings suggest that the impaired glucose metabolism is a significant risk factor for coronary atherosclerosis in Japan. The clinico-pathological survey of risk factors for atherosclerosis in the general Japanese population would be very important in not only the further clarification of Japanese atherogenesis but also the exploration and development of novel diagnostic and therapeutic strategies, and preventive guidelines for atherosclerosis-related cardiovascular diseases in Japan nowadays under the remarkable changes of life-style. PMID- 17087291 TI - [Epidemiology of macroangiopathy in diabetes, including international comparisons]. AB - Individuals with diabetes have an increased risk of cardiovascular disease. Epidemiological studies demonstrate that plasma glucose (PG) concentrations in the upper normal range are an independent risk factor for cardiovascular disease, as discussed in a meta-analysis. Studies carried out mainly in the general population show that post challenge PG predicts the incidence of cardiovascular events and mortality more than fasting PG: however, results obtained measuring PG after an oral glucose tolerance test cannot be extrapolated to the postprandial condition. Two studies, the Diabetes Intervention Study (DIS) and San Luigi Gonzaga Diabetes Study, indicated that postprandial PG predicted cardiovascular events in diabetes. PMID- 17087292 TI - [Long-term intervention studies of diabetic macroangiopathy in Japan and western countries]. AB - Diabetes is a strong and independent predictor of cardiovascular disease that seriously affects mortality and morbidity of patients. Risk of macroangiopathy starts to increase from very early stages of diabetes compared to microangiopathy, therefore aggressive and early treatment is necessary to reduce its risk. Results of previous intervention studies suggest that treatment of each single risk factor is somewhat effective but not enough to achieve sufficient protection from macroangiopathy events, therefore simultaneous interventions of many cardiovascular risk factors are required. At this moment, intervention studies on East Asian patients are very limited and extensive future investigations are expected. PMID- 17087293 TI - [Coronary heart disease in patients with diabetes mellitus]. AB - Diabetes mellitus is a well-recognized risk-factor for coronary artery disease. It is known that diabetic patients have a two-to fourfold increased risk of mortality or morbidity for cardiovascular diseases compared with the general population. Further, many diabetic patients with coronary heart disease have severe diseased coronary arteries: multi-vessel disease and/or severe stenosed vessel(s). In Japan, the prevalence of diabetic patients is rapidly increasing in these days. This article will review the evaluation for risk of coronary heart disease by measuring glucose level, the prognosis of diabetic patients, and the prevention of cardiovascular event in the patients with diabetes mellitus. PMID- 17087294 TI - [Cerebral infarction]. AB - Diabetes mellitus is the second major risk factor for ischemic stroke. Recent increase in atherothrombotic stroke appears to be related with recent increasing of diabetes. Diabetes is, however, a risk factor not only for atherothrombotic stroke but also for lacunar stroke because there is no difference in prevalence of diabetes between atherothrombotic and lacunar strokes. Diabetes can be a risk factor for cardioembolic stroke as well because the major cause of cardioembolic stroke is atrial fibrillation, and diabetes is a risk factor for stroke in patients with atrial fibrillation. Acute ischemic stroke should be classified into above three subtypes according to the brain and artery imaging as well as cardiac sources of embolism. In hyper-acute patients within 3 hours of onset and without early ischemic signs on CT or ischemic lesions less than one third of the hemisphere on magnetic resonance diffusion-weighted imaging, thrombolytic therapy with alteplase is indicated. In acute stroke patients later than 3 hours of onset, argatroban, heparin, and ozagrel are indicated for atherothrombotic, cardioembolic, and lacunar stroke, respectively. For stroke prevention, total management is required by simultaneous treatments for all risk factors existed. In secondary prevention for stroke, in addition to the more strict control of risk factors antithrombotic therapy is required, that is, antiplatelet therapy is indicated for non-cardioembolic stroke, and anticoagulant therapy is indicated for cardioembolic stroke. PMID- 17087295 TI - [Overview on diagnosis, treatment and therapeutic angiogenesis for arteriosclerosis obliterans]. AB - Arteriosclerosis of the extremities is a disease of the blood vessels characterized by hardening and/or narrowing of the arteries that supply the legs and feet. This causes a decrease in blood flow that can injure nerves and other tissues. Therapeutic angiogenesis using angiogenic growth factor is expected to be a new treatment for patients with critical limb ischemia. The first human clinical trial treating peripheral vascular disease was started in 1994 using vascular endothelial growth factor. To date, other potent angiogenic growth factors, such as hepatocyte growth factor(HGF), have been also estimated in clinical trials for peripheral arterial disease. Several results from phase 1 or 2 trials using HGF gene were encouraging. Phase 3 trials are now ongoing and their results are expected. PMID- 17087296 TI - [Current status and perspectives of coronary imaging for patients with diabetes mellitus]. AB - Diabetes mellitus (DM) is an independent and distinct risk factor of ischemic heart disease. Therefore, early identification and management of coronary atherosclerosis for patients with DM is being desired. For the purpose of visualizing coronary atherosclerosis, various imaging modalities have been proposed especially to identify an unstable plaque that has a thin fibrous cap and a large lipid core. These modalities include intravascular ultrasound, coronary angioscopy, intracoronary thermography, optical coherence tomography, multi-slice CT, MRI, and so on. The development of these modalities are now pushing all cardiologists to get interested in a new field called as "plaque imaging". PMID- 17087297 TI - [New technique of magnetic resonance imaging to evaluate peripheral circulation in diabetic patients]. AB - Waveform analysis at the popliteal artery using the new technique of gated two dimensional cine-mode phase-contrast magnetic resonance imaging is beneficial to assess peripheral circulation in both normal and diseased arteries. The normal subjects had a typically triphasic waveform, which could be clearly separated into systolic, and early and late diastolic phases of the cardiac cycle. Diabetic patients are reported to have two types of insufficient arterial blood flow to the lower limbs associated with the vessel wall properties. Firstly, patients with atherosclerotic occlusion in the lower-leg arteries distal to the aortic bifurcation, resulting in a low ankle-brachial index, show an abnormal monophasic waveform. Secondly, patients with stiffer arteries characteristically show abnormal flow reversal in late diastole, suggesting higher vascular resistance. PMID- 17087298 TI - [Methods to evaluate the vascular endothelial function]. AB - When the vascular endothelial function fails, arterial atherosclerosis is developed. Furthermore, if the endothelial dysfunction progresses, it is known to cause angina pectoris, myocardial infarction, cerebrovascular accident. By medical therapy and replacement therapy, correction of the life style, the endothelial dysfunction is understood that it could be improved reversibly. Therefore, it is very important to evaluate the vascular endothelial function for arteriosclerotic severity and the prediction of the cardiovascular disease onset and the effect of medical therapy. In the 1990s, high-frequency ultrasonographic imaging of the brachial artery to assess endothelium-dependent flow-mediated vasodilation (FMD) was developed. This technique is attractive because it is noninvasive and allows repeated measurements. However, despite its widespread use, there are technical and interpretive limitations. Therefore, for clinical application of the endothelial function test, its problems must be understood well. PMID- 17087299 TI - [Guidelines for the management of atherosclerotic diseases in diabetes mellitus]. AB - Diabetes mellitus is associated with a greater risk of developing atherosclerosis and its complications: myocardial infarction, stroke and peripheral vascular disease. In patients with diabetes, atherosclerosis represents a complex multifactorial disease with increased lesion progression and severity compared to the nondiabetic population. Several risk factors have been proposed to explain the increased risk of cardiovascular disease with diabetes. They include: hyperglycemia, hypertension, dyslipidemia, obesity and other factors. It is difficult to precisely establish the elements leading to diabetes-accelerated atherosclerosis by means of epidemiological studies because all these factors coexist in diabetic patients. Then, management of atherosclerosis in diabetes is a multifactorial process involving nonpharmacological interventions like exercise, diet control, and pharmacological therapy directed at hypertension, hyperglycemia, and dyslipidemia. PMID- 17087300 TI - [Lifestyle to prevent cardiovascular disease in NIDDM]. AB - Major risk factors for cardiovascular diseases (myocardial infarction, angina pectoris, brain infarction) are age, male, smoking, high LDL-cholesterol, low HDL cholesterol, high blood pressure, and diabetes mellitus. In Japanese population at large, healthy life-style to prevent cardiovascular diseases are; quit smoking, walking faster, saturated fat intake ranging 4.5-7 en%, lesser intake of trans fatty acids, cholesterol intake less 750 mg/day (male) and 600 mg/day (female), eat fish everyday, eat more folic acid, B6, and B12, eat grain, eat soybean products. However, it is not known whether this recommendation is also applied to NIDDM to prevent cardiovascular diseases. Based on reported evidences, to prevent cardiovascular diseases, NIDDM should quit smoking, eat fish everyday, and increase physical activity. PMID- 17087301 TI - [Management of blood glucose--target of the management and its practice]. AB - Accumulating evidence supports that management of blood glucose levels is critical for the prevention of diabetic vascular complications including macroangiopathy. Several therapeutic modalities are available in the management of blood glucose levels. In type 2 diabetes with obesity, achievement of body weight control with intensive caloric restriction is essential. Sulfonylurea remains the central drug in the management in insulin-independent patients without obesity. For patients with insulin resistance, biguanide or thiazolidinediones are the drugs of choice. For patients with postprandial hyperglycemia, a-glucosidase inhibitors and meglitinide analogs should be considered. Combination of some of these drugs has additive effect for patients with suboptimal blood glucose control with monotherapy. When the above therapies fail, we should not hesitate to start insulin. PMID- 17087302 TI - [Treatment for hypertension with diabetes mellitus]. AB - In the treatment for the hypertension patients with diabetes mellitus, the target blood pressure is below 130/80mmHg. As the first choice drugs, ARB, ACE inhibitor and Ca channel blocker are recommended. In the high normal blood pressure patient (130-139/80-89mmHg), the initial approach is life-style modification. In hypertension patients with diabetes (over 140/90mmHg), blood pressure should be controlled with the medication. The life-style modification should be performed even in the patients treated with anti-hypertensive drugs. PMID- 17087303 TI - [Lipid management--treatment goal and strategy]. AB - Diabetic macroangiopathy is known to increase with complicated hyperlipidemia. Large clinical studies indicate that statins are effective in reducing cardiovascular events in subjects with type 2 diabetes. Target goal for LDL cholesterol in type 2 diabetes is less than 100 mg/dl in the United States, whereas it is less than 120mg/dl in Japan. Aggressive lipid-lowering therapy is indicated for type 2 diabetes, especially those with high risk. Other effects than deceasing LDL of statins, i.e. pleiotropic actions, are suggested to contribute to reduction of cardiovascular events. A decrease in HDL cholesterol and an increase in triglycerides are also risk factors for cardiovascular events, but some portion of them are attributed to dyslipidemia associated with insulin resistance. PMID- 17087304 TI - [Glinide(s), sulfonylurea(s)]. AB - Diabetic macroangiopathy has already developed before diagnosis of diabetes mellitus. Postprandial hyperglycemia has been known as a risk factor for diabetic macroangiopathy and may be more powerful than fasting hyperglycemia. To intervene in hyperglycemia, insulin secretagogues, glinides which selectively stimulate early meal-induced insulin secretion and improve postprandial hyperglycemia, and sulfonylureas which enhance total daily insulin secretion and improve fasting hyperglycemia, have been prescribed as major oral antidiabetic agents. Few evidences that amelioration of glycemic control with insulin secretagogues lower the risk of cardiovascular diseases have been reported. But current studies have shown that intervention in postprandial hyperglycemia with drugs including glinides decreased thickness of carotid IMT as a surrogate marker of atherosclerosis. Results from on-going large scale intervention study with glinides may clarify whether amelioration of hyperglycemia lower the risk of atherosclerotic events. PMID- 17087305 TI - [Statins (HMG-CoA reductase inhibitors)]. AB - Diabetic patients frequently show lipid abnormalities that include qualitative changes in lipid profile. Given the greatly increased risk of coronary heart disease (CHD) in diabetic patients with hyperlipidemia, most principal clinical guidelines recommend aggressively lowering lipid levels in such patients. In fact, several large-scale clinical intervention trials have successfully shown that lipid-lowering agents, particularly statins, could significantly reduce the CHD risk in diabetic patients with hyperlipidemia. According to our investigation on 9,000 Japanese diabetic patients, about a half had hyperlipidemia but only less than half of the hyperlipidemic patients received some lipid-lowering agents. Taken together, it should be concluded that more stringent and aggressive intervention should be recommended in the treatment of Japanese diabetic patients with hyperlipidemia. PMID- 17087306 TI - [Prevention and treatment for development and progression of diabetic macroangiopathy with pioglitazone and metformin]. AB - Prevention and treatment for diabetic macroangiopathy, causing the main etiology for mortality in patients with diabetes mellitus, is crucial target point, but is still controversial. Many clinical studies improving glycemic control by insulin and oral drugs were not demonstrating enough prevention for diabetic macroangiopathy to improve life prognosis. Insulin resistance is now considered to be major pathogenesis for diabetic macroangiopathy. The agents that improve insulin resistance, such as metformin and pioglitazone, have multiple effects to improve glucose and lipid metabolism by increasing sensitivity for insulin without increasing insulin secretion and exert anti-atherogenic properties resulting in preventing development of atherosclerosis. Some clinical studies such as UKPDS 34 and PROactive demonstrated preventive effects of these agents for diabetic macroangiopathy. PMID- 17087307 TI - [Therapeutic angiogenesis for peripheral arterial disease and ischemic heart disease by autologous bone marrow cells implantation]. AB - Postnatal neovascularization was initially thought to result exclusively from the migration and proliferation of pre-existing, fully differentiated endothelial cells (a process referred to as angiogenesis). Several lines of experimental evidence, however, demonstrated that circulating endothelial progenitor cells (EPCs) home to sites of neovascularization and differentiate into endothelial cells in situ in a manner consistent with a process termed vasculogenesis. Moreover, recent clinical study, named as the Therapeutic Angiogenesis Using Cell Transplantation (TACT), has demonstrated the efficacy and safety of autologous implantation of bone marrow-mononuclear cells(BM-MNCs) in randomly allocated patients with ischemic limbs of peripheral arterial diseases. Implantation of BM MNCs is also applied to patients with no-option angina pectoris, demonstrating the usefulness of this therapy for improvement of symptom, myocardial perfusion, and exercise capacity. Moreover, recent accumulating studies demonstrated the possibilities that transplantation of bone marrow-derived or peripheral blood derived EPCs enhances neovascularization and improves cardiac function after acute myocardial infarction(AMI). There were no serious adverse effects reported in cell transplantation therapy. Thus, bone marrow-derived cell transplantation therapy has been applied to acute and chronic ischemic heart diseases as well as peripheral arterial diseases, demonstrating the safety and effectiveness of therapeutic angiogenesis. PMID- 17087308 TI - [Vascular endothelium regeneration therapy]. AB - CD34 positive cells were first defined as endothelial progenitor cells(EPCs) from circulating mononuclear cells in peripheral blood. EPCs have shown to be mobilized from bone marrow by the various factors, incorporate into sites of physiological and pathological neovascularization and differentiate into mature endothelial cells (ECs). Post-natal vasculogenesis has been considered to be involved in neovascularization of adult tissues. Recently, freshly isolated CD34 positive cells transplantation has started as clinical trial for ischemic diseases. In the clinical situation, we should consider the cell number and cell quality derived from the patients who have atherosclerosis background, especially diabetes. Ex vivo expansion or gene modification of EPCs could be the strategies for the next generation cell therapy to overcome these issues. PMID- 17087309 TI - [Regenerative medicine with the sustained release system of basic fibroblast growth factor]. AB - Recently we have developed new sustained release system of basic fibroblast growth factor (bFGF) using gelatin hydrogel as a carrier biomaterial. We examined the effect of topical sustained release of bFGF on blood flow recovery and tissue regeneration in various animal models of cardiovascular area. These results revealed effectiveness and safety of this therapy. We evaluated the effect of sustained release of bFGF for not only normal but also diabetes model. Slow release system of bFGF from biodegradable gelatin hydrogel prevents bFGF glycation and its combination with 5-hydroxytryptamine blocker has shown sufficient neovascularization in diabetic limb ischemia. The method may provide a more effective therapeutic angiogenesis in patients with diabetes. Clinical trial of therapeutic angiogenesis for severe hindlimb ischemia has already started. PMID- 17087310 TI - [Strategy for order-made treatment of diabetic macroangiopathy]. AB - Several genetic risk factors such as single nucleotide polymorphisms(SNPs) in candidate genes have been reported to be responsible for diabetic macroangiopathy. We determined their genotypes regarding more than 100 SNPs in candidate genes responsible for average IMT of Japanese type 2 diabetic patients. We depicted some SNPs which were found to be significantly (p<0.05) responsible to the increase in AveIMT. Then, two combinations of double SNPs were evaluated as responsible for significant increases in AveIMT. Also, these two combinations were probably responsible for a high frequency of history of old myocardial infarction. The present analysis may provide one approach to evaluate combination of multiple genetic risk factors, which is synergistically associated with carotid atherosclerosis and myocardial infarction. PMID- 17087311 TI - [Homocysteine and diabetic macroangiopathy]. AB - Moderate hyperhomocysteinemia is one of risk factors for arteriosclerotic disease. In diabetic patients, hyperhomocysteinemia is an independent risk factor for macroangiopathy and mortality. Homocysteinemia is also associated with diabetic microangiopathy, silent stroke, and cognitive impairment. However, excluding those with nephropathy or microangiopathy, plasma homocysteine is lower in diabetic patients than non-diabetic controls. Oral treatment with folic acid, vitamin B12 and B6 reduces plasma homocysteine concentration about by 30%. The vitamin treatment for reduction of hyperhomocysteinemia improves endothelial dysfunction and retards carotid atherosclerosis. Few randomized control trials have showed a positive effect of the vitamin treatment on prevention from stroke and ischemic heart disease. Further prospective intervention studies are necessary to address the issue whether lowering homocysteine does prevent the development and progression of diabetic macroangiopathy. PMID- 17087312 TI - [Biorheological views of diabetic macroangiopathy]. AB - Diabetes mellitus markedly increases the risk of myocardial infarction, stoke, amputation which cause most morbidity and mortality. Elevated low shear blood viscosity with erythrocyte aggregation and elevated high shear blood viscosity with reduced erythrocyte deformability might be important and potentially treatable factors in the etiology or progression of diabetic macroangiopathy. The metabolic abnormalities caused by diabetes induce both rheological changes of blood and vascular dysfunction that predisposes this patient population to atherosclerosis. Lowered shear stress and vortices with oscillatory shear stress induce atherosclerosis. Abnormalities in platelet function and increased blood coagulability may exacerbate the progression of atherosclerosis and the consequences of plaque rupture or erosion in diabetes. PMID- 17087313 TI - [Primary care in pediatrics: Immediate diagnosis in first visit]. PMID- 17087314 TI - [NEMOes, a new method to evaluate early atherosclerosis in rodent model]. AB - We recently established new en face method for optimal observation of endothelial surface(NEMOes) that enabled us to obtain clear images of the entire endothelial surface, including at arterial bifurcations, and to quantitate the number of cells of interest in the rodent endothelium. Using this method, we found that not only advanced glycosylation endproducts, but also glucose spike induced a significant increase in the number of macrophages adhering to the endothelium. NEMOes may be a promising tool for understanding the pathophysiology of atherosclerosis. PMID- 17087315 TI - [Social epidemiology: the development and prospect]. PMID- 17087316 TI - [Physical activity level and physical, mental and social factors in community dwelling elderly people]. AB - PURPOSE: This study was conducted to examine the relationship between physical activity level (PAL) and physical, mental and social factors in community dwelling elderly people. METHODS: The subjects comprised 428 residents aged 70 or over living in Kusatsu-machi, Gunma, who took part in an interview and tests of physical fitness as part of the "NIKKORI-KENKOSODANJIGYO". Data for 330 out of the 428 residents were adopted for this study. The PAL was evaluated with a questionnaire developed by Naito et al. (2003). Subjects were interviewed on physical, mental and social functioning including the Tokyo Metropolitan Institute of Gerontology (TMIG) Index of Competence and cognitive function (Mini Mental State Examination: MMSE). Physical fitness tests included assessment of handgrip strength, usual and maximal walking speed, and one-leg standing balance with eyes open. RESULTS: Correlations between PAL and the TMIG Index of Competence, physical (e.g. physical fitness), mental (e.g. depression) and social (e.g. roles at home) factors, and smoking status were significant by ANCOVA adjusted for age and sex. Analysis using a general linear model indicated that smoking status, usual walking speed, depression, roles at home, frequency of going outdoors and visual impairment all together explained 13.5% of the PAL variance. CONCLUSION: The findings indicate that PAL in the community-dwelling elderly aged 70 years or over is associated with physical, mental and social factors. PMID- 17087317 TI - [Social interaction and mortality: a seven-year longitudinal study of elderly people]. AB - OBJECTIVES: This study was designed to clarify the relation between social interaction and seven-year-mortality in a group of elderly people. METHODS: The participants were all 65 years or older living in farming communities near major urban centers in Japan (n = 801). The contents of the questionnaire covered social interaction (using an "Index of Social Interaction" consisting of 18 items), health status, physical function, activities of daily living (ADL), age and gender. RESULTS: A total of 139 subjects died within the seven year period after the baseline survey: 1) baseline age and physical function were related to the seven-year mortality; 2) greater social interaction was positively related to reduced mortality; 3) multiple logistic regression analysis adjusted for baseline age, gender, physical function, health status, and ADL indicated that greater social interaction was related to reduced mortality. DISCUSSION: These findings highlight the importance of social interaction in successful aging. PMID- 17087318 TI - [Longitudinal analysis of factors associated with participation in community based mass screening for the frail elderly in need of care]. AB - PURPOSE: To examine factors associated with participation in a community based comprehensive health check to screen frail elderly before they become in need of long-term care. METHODS: All residents aged 70 years and over living in Kusatsu, Gumma prefecture were surveyed for their sociodemographics, and physical, mental and social functioning through in-person interview in the years 2001 and 2003. In the following years--in 2002 and 2004, respectively--mass screenings were conducted to detect early signs of need for care among the elderly. Using the baseline interview information as explanatory variables, we performed multiple logistic regression analysis in order to examine the social determinants of participation in the mass-screening. RESULTS: For the first mass-screening, having high blood pressure and poor subjective health decreased the likelihood of attending the screening by 34% and 65% respectively. In contrast, one point increases in IADL, mobility and social-role score increased the probability of attendance by 27%, 26% and 26% respectively. After the screening was re conducted, the factors influencing attendance appeared to have changed. With a third mass-screening, while the IADL and mobility score still had significant effects, elderly having excellent subjective health were less likely to go by some 48%. It was also found that not having visual impairment had a significantly positive effect on attendance. CONCLUSION: If local government conducts community based mass screening aiming to screen the frail elderly in need of care, it is likely that there are high risk elderly among the non-participants. It is very important to follow up those non-participants using an attendance roll and provide appropriate advice. It is also worth discussing mass screening that focuses more on health promotion than on "screening" itself. PMID- 17087319 TI - [An intergenerational health promotion program involving older adults in urban areas. "Research of Productivity by Intergenerational Sympathy (REPRINTS)": first year experience and short-term effects]. AB - OBJECTIVE: We have launched a new intervention study, called "Research of Productivity by Intergenerational Sympathy (REPRINTS)" in which senior volunteers engage in reading picture books to children. The "REPRINTS" program consistently involves social roles and intellectual activity, two higher-level functional capacities. This study reported findings and problems experienced through "REPRINTS" during the first year, ascertained potential effectiveness of social activity, and proposed methods for continued activity. METHODS: Basic concepts of "REPRINTS"program include "contribution to society", "life-long learning", and "group activity." Sixty seven volunteers and 74 controls, all aged 60 years and over living in three areas, ie., Chuo-ku, central Tokyo, Kawasaki city, suburb of Tokyo and Nagahama city, a local city, participated in a baseline health check-up in June, 2004. After completion of 3-month training seminars (once a week, 2 hr per session), volunteers visited public elementary schools and kindergartens in groups of 6-10 persons for 6 months. They were assessed again by follow-up health check-up in March, 2005. RESULTS: At baseline, the proportion of those who had no grand children (41.8% vs. 20.3%, P= 0.006), average school years (13.4 +/- 2.5 vs. 12.3 +/- 2.5 years, P= 0.008), having any experience of volunteer activities (79.1% vs. 52.7%, P=0.001), and an usual walking speed (86.7 +/- 12.3 vs. 81.3 +/ 12.9 m/min, P=0.012) were significantly higher in volunteers than in controls. There was no significant difference in other baseline characteristics between the two groups. At follow-up, social network scores for 56 volunteers were significantly improved: frequency of contact with grandchildren and others around neighborhood and size of circles of friends and acquaintances were increased, as compared to controls. Social support scores for the volunteers significantly decreased in the receiving aspect, while increased in the giving aspect. In addition, consciousness of loving community and self-rated health were significantly enhanced and hand-grip strength showed significantly less decline in volunteers than in controls. CONCLUSIONS: Through engaging in the intergenerational and intellectual volunteer programs for 9 months, subjective health, social support and network, consciousness of loving community and some aspects of physical performance were significantly promoted in senior volunteers. Coordination of existing administrative services enabled us to establish this "REPRINTS" health promotion program for community-dwelling older adults. PMID- 17087320 TI - [A qualitative study on effective postgraduate clinical training of public health and community health care]. PMID- 17087321 TI - Initial clinical experience with the Heartstring. AB - OBJECTIVE: The Heartstring is one of the devices that enable proximal anastomosis without clamping the aorta. We have applied the device not only to low-risk patients with normal aortas but also to high-risk patients with diseased aortas. The purpose of this study was to investigate the initial outcomes of using this device. METHODS: The Heartstring was used on 87 patients between January and December 2004. The patients comprised 62 men and 25 women 48-86 years old (mean 68.4 +/- 8.4 years). The ascending aorta was evaluated by computed tomography (CT) scanning before surgery. If a patient's aorta was severely calcified, epiaortic echocardiography was performed. The aortas were ranked into four grades, and the preoperative patient's status were evaluated by the EuroSCORE. Angiography was performed on the third postoperative day. RESULTS: CT scanning revealed that 74 patients had no calcification in the ascending aorta, 10 patients had scattered calcification, and 3 patients had plate-like calcification. The EuroSCORE was 6.86 +/- 1.03. We performed 93 proximal anastomoses and 149 distal anastomoses. The average distal anastomosis was 1.6 +/ 0.6 sites per graft. Of the distal anastomoses, 74.2% were to the circumflex artery territory. Postoperative coronary angiography revealed that all grafts were patent. CONCLUSION: The Heartstring facilitates safe proximal anastomosis, even in high-risk patients. Their short-term outcome was good. The device assists in bypassing circumflex artery territory. Long-term follow-up is necessary. PMID- 17087322 TI - Surgical repair of the common arterial trunk in a neonate with a downsized valve allograft. AB - In the present report we describe a neonate with the common arterial trunk who underwent reconstruction of the right ventricular outflow tract with a downsized value allograft. Our experience and results suggest that reducing the size of the allograft is a promising surgical option for young infants in whom reconstruction of the nonregurgitant pulmonary circulatory system is absolutely needed for a smooth postoperative recovery. PMID- 17087323 TI - Giant intraatrial septal aneurysm originating from a branch of the left coronary artery. AB - We present the case of a 38-year-old woman who had a large intraatrial aneurysm occupied by old thrombosis. The aneurysm was successfully removed, and the atrium was repaired. Pathohistological findings indicated that the inflow artery of the aneurysm had an anomalous origin from the left main coronary artery, and its pathogenesis was unknown. It is occasionally difficult to distinguish a large coronary aneurysm from a mediastinal tumor because this aneurysm is a rare entity, even more so in an atrial septum. A giant coronary aneurysm should be considered an alternative diagnosis in the event of a mediastinal mass. Surgery is recommended for a large coronary aneurysm. PMID- 17087324 TI - Intralobar sequestration presenting as a large intrapulmonary hematoma and massive hemothorax. AB - Intralobar sequestration is a relatively rare anomaly that is usually diagnosed with symptoms of cough, expectoration, or recurrent pneumonia. We experienced a case of a 27-year-old man with a symptom of massive hemoptysis. His chest computed tomography (CT) scan revealed a large intrapulmonary hematoma and massive hemothorax, mimicking a benign lung tumor ruptured into the pleural cavity. We should keep the possibility of this anomaly in mind if a patient with hemoptysis has a cystic lung tumor and hemothorax on CT scan. PMID- 17087325 TI - Transudative pleural effusion associated with left brachiocephalic vein resection. AB - A 70-year-old woman was admitted for surgical treatment of a thymoma. She underwent thymectomy with left brachiocephalic vein resection because of invasion. Left pleural effusion appeared from postoperative day 4 and resolved with drainage. Herein, we report a rare case of pleural effusion associated with brachiocephalic vein resection. PMID- 17087326 TI - Multiple thoracoabdominal gunshot wounds with complicated trajectories. AB - A 41-year-old man was shot three times with a custom-made handgun. There were six small round wounds on the surface from two bullets that went into the body and one bullet found in a wall behind the victim at the scene. The gunshot wound in the left supraclavicular region bled massively. He was transferred to our hospital and soon fell into shock. A chest radiogram showed right hemothorax. Resuscitation and blood transfusion were started, and right thoracic drainage was immediately undertaken. The left subclavicular region was incised, and bleeding from the left subclavian vein stopped. Right thoracotomy was done, and bleeding from the lung was controlled. Laparotomy was needed to look for the missing trajectory. Speculation regarding trajectories of bullets in patients with multiple thoracoabdominal gunshot wounds is difficult, and an emergent operation is often needed to control bleeding or to evaluate possible organ injury. PMID- 17087327 TI - Redo total aortic arch replacement using an extended homograft for graft infection. AB - A 65-year-old man developed graft infection 3 years after total aortic arch replacement for a chronic dissecting aneurysm. Redo total aortic arch replacement was performed using an extended homograft following 2 days of open drainage of the mediastinum. The patient has been free from a recurrent mediastinitis over the last 3 years. PMID- 17087328 TI - Recurrent discrete subaortic stenosis and small aortic annulus successfully repaired by the Konno procedure in a young woman. AB - A 24-year-old woman who had undergone excision of the membrane for discrete subaortic stenosis when 6 years old displayed recurrent subaortic stenosis and had a small aortic annulus, with a peak pressure gradient of 60mmHg. We chose to perform the Konno operation with a mechanical valve. This released the left ventricular outflow tract obstruction adequately, and she recovered uneventfully with New York Heart Association functional class I. In our experience, an aggressive strategy such as myectomy is an appropriate initial procedure for preventing recurrence when the geometry of the problem may lead to recurrence in the left ventricular outflow tract. The Konno operation is a good option for recurrent subaortic stenosis with small aortic annulus. PMID- 17087329 TI - On-pump beating-heart mitral valve plasty without aortic cross-clamping. AB - Patients with ischemic cardiomyopathy often have mitral regurgitation, which should be corrected for better long-term survival. Mitral valve surgery is usually performed during cardiopulmonary bypass under the arrested heart condition. The ascending aorta is cross-clamped and the heart is arrested using a cardioplegic solution. However, because ischemic cardiomyopathy patients often have a severely atherosclerotic ascending aorta and low cardiac function, aortic cross-clamping and cardiac arrest increase the risk of postoperative thromboemboli and low cardiac output syndrome. Under the on-pump beating-heart condition, we performed mitral valve plasty concomitant with coronary artery bypass grafting, tricuspid annuloplasty, left ventricular aneurysmectomy, and the maze procedure without aortic cross-clamping for a patient with ischemic dilated cardiomyopathy and bradycardial atrial fibrillation. The patient had no postoperative complications and re covered rapidly. Thus, to prevent serious postoperative complications, on-pump beating-heart mitral valve surgery without aortic cross-clamping may be a suitable surgical option for patients with ischemic cardiomyopathy. PMID- 17087330 TI - Malignant pleural mesothelioma in a 14-year-old boy with right aortic arch. AB - We report successful surgery for malignant pleural mesothelioma (MPM) in a 14 year-old boy with right aortic arch. Pleural biopsy by video-assisted thoracic surgery yielded a diagnosis of MPM, epithelial type. As the disease was not changed after combination chemotherapy with three cycles of cisplatin and gemcitabine, we performed left extrapleural pneumonectomy, including resection of the pericardium and diaphragm for MPM, and aortopexy for right aortic arch. The postoperative course was uneventful, and the patient has remained alive without disease for 10 months postoperatively. PMID- 17087331 TI - [Surgical lung biopsy for interstital lung diseases: complications, diagnostic yield and mortality]. AB - We retrospectively reviewed 110 consecutive patients with interstitial lung diseases (ILDs) who underwent surgical lung biopsy over 13 years from 1989-2002. Surgical procedures, medical records, surgical complications and outcome including post-operative exacerbation of ILDs were assessed in order to clarify the safety and efficacy of surgical lung biopsy. Forty-one patients underwent open lung biopsy (OLB), while 69 patients received videothoracoscopic lung biopsy (VTLB). There was no significant difference in patient characteristics, pulmonary function, arterial oxygen pressure and therapy prior to surgery between the two groups. The mean number of biopsy specimens was 2.5. Left side was selected as the site of biopsy in 70 cases. Duration of surgery was not significantly different between 2 groups. Bleeding during operation was less in the VTLB group than in the OLB group, although the mean bleeding amount was only 16.5mL. The duration of thoracic drainage and length of hospital stay were shorter in the VTLB group. The overall rate of surgical complications was 10.9%, with no difference between the two groups. The overall 30-day post-SLB mortality was 6.4% and majority of these had a very poor prognosis pathologically. The diagnostic yield of all 110 cases was 98.2%. SLB is a relatively safe and efficient diagnostic method in the diagnosis of ILDs. PMID- 17087332 TI - [Clinical features and outcome in 84 patients with influenza pneumonia]. AB - We examined the clinical features, illness types of pneumonia, efficacy of neuraminidase inhibitors and outcome in patients with influenza pneumonia. Eighty four patients with influenza pneumonia, in whom the diagnosis was confirmed by serology or rapid diagnostic tests, were studied. Because neuraminidase inhibitors were given prior to the onset of pneumonia in some patients with secondary bacterial pneumonia, we examined the efficacy of neuraminidase inhibitors in patients other than the secondary bacterial pneumonia group. Influenza A was detected in 71% of the subjects, and influenza B in 29% who showed a clinical presentation and outcome similar to those patients with influenza A. Primary influenza viral pneumonia was observed in 27% of the subjects, mixed viral and bacterial pneumonia in 38%, secondary bacterial pneumonia in 18%, and unclassified type of pneumonia in 17%, respectively. Eighty three percent of the subjects had underlying diseases. The overall mortality rate was 9.5%, and all 8 fatal cases had comorbidities. Thirty-nine percent were aged under 65 years and 3 fatal cases were aged under 65 years. The mortality rates of patients receiving or not receiving neuraminidase inhibitor were 4.9% and 14.3%, respectively. Due to the small number of patients and the fact that this case was not a controlled study, the efficacy of neuraminidase inhibitors for influenza pneumonia was not proved. PMID- 17087333 TI - [Autopsy case of sarcomatoid malignant pleural mesothelioma]. AB - A 61-year-old man with a sensation of chest compression was admitted to our hospital. He had hemothorax. After drainage with a chest tube, chest CT scan revealed multiple bilateral pulmonary nodules with slight pleural thickening. Open pleural biopsy was performed and the biopsy specimens showed tumor cells with sarcomatoid proliferation, but no definite epithelial pattern. Initial immunohistochemical staining was negative for keratin and carletinin, but positive for desmin, suggesting rhabdomyosarcoma. After supportive care, he died due to progression of the disease. Autopsy revealed extensive invasion suggesting mesothelioma, so the immunohistochemical staining was repeated. Because it revealed patchy staining for keratin and carletinin, this case was diagnosed as sarcomatoid mesothelioma. Differential diagnosis of sarcomatoid mesothelioma or rhabdomyosarcoma is made by immunohistochemical staining, but it is sometimes difficult. For the selection of the best treatment strategy for mesothelioma especially in the early stage, we should be aware of this difficulty. PMID- 17087334 TI - [Case of severe eosinophilic pneumonia, induced by several antibiotics, requiring mechanical ventilation]. AB - We report a case of severe eosinophilic pneumonia induced by several antibiotics, requiring mechanical ventilation. A 44-year-old man had been admitted previously to a different hospital because of a high fever and dry cough continuing for a week. Although bacterial pneumonia was diagnosed and he had been treated with several antibiotics, his respiratory state worsened gradually and he was refered to our hospital with severe hypoxia. He needed mechanical ventilation on the fifth hospital day. TBLB identified eosinophilic pneumonia. Although we discontinued all antibiotics and started steroid therapy, he became feverish and his leukocyte count, which was neutrophil dominant, elevated again. We suspected bacterial pneumonia and administered antibiotics that he had never received before. However we had to discontinue these antibiotics soon because of a further inflammatory response. His condition then improved gradually and he was weaned from mechanical ventilation. It is rare for drug-induced eosinophilic pneumonia to become so severe that mechanical ventilation is necessary. We thought that an allergic reaction may have been induced by the series of several antibiotics, inducing severe pneumonia. PMID- 17087335 TI - [Case of bronchial asthma complicated with Takotsubo cardiomyopathy after frequent epinephrine medication]. AB - A 62-year-old man with non-topic severe persistent asthma and chronic obstructive disease suffered severe asthma exacerbation. Epinephrine was repeatedly injected subcutaneously (0.3mg x 8 times in four hours) in addition to intravenous methylprednisolone for his severe asthma. Despite these treatments, his symptoms steadily deteriorated and thus he was transferred to our hospital. He did not have chest pain or a sensation of compression, while ECG on admission demonstrated ST elevation, loss of R-wave progression, negative T waves and QT interval prolongation, suggesting ischemic heart disease. Nonetheless creatine kinase and its MB isozyme were within normal range and myosin light chain I and troponin T were only mildly elevated. Echocardiography demonstrated apical dyskinesia and hypersystole in the basal region of the heart. Finally this case was diagnosed as Takotsubo cardiomyopathy, probably due to catecholamine-mediated myocardial stunning by overuse of epinephrine for acute severe asthma exacerbation. Abnormal findings of ECG and echocardiography became normal without any specific treatments for the heart. PMID- 17087336 TI - [Case of lung cancer with chylothorax]. AB - A 42-year-old man had swelling in the right side of the neck, cough and chest pain. On admission, an abnormal shadow was detected in the right upper lung field and squamous cell carcinoma of the lung with superior vena cava (SVC) syndrome was diagnosed. Concurrent radiotherapy and systemic chemotherapy consisting of cisplatin and vinorelbine induced a partial response. At 15 months after diagnosis, he was re-admitted because of bilateral pleural effusion and facial edema due to relapse of SVC syndrome. Examination of the milky right pleural effusion revealed chylothorax (959mg/dl of beta-lipoprotein and 675mg/dl of triglyceride). The right effusion was finally controlled by pleurodesis with OK 432. Non-traumatic chylothorax is a rare complication of lung cancer. PMID- 17087337 TI - [Case of cavitary coccidioidomycosis with fungus balls in the apices of both lungs]. AB - Pulmonary cavitary coccidioidomycosis with a fungus ball was observed in a immunocompetent case. A 32-year-old Japanese man visited Arizona to play golf. After 1 month he consulted a local hospital complaining of a prolonged cough and hematopysis. The laboratory examination revealed eosinophillia and chest radiograph showed 2 cavitary lesions, surrounded by small nodules in the apices of both lungs. Pulmonary tuberculosis was suspected and treated with 4 antituberculosis drugs for 3 months. However, the cavities enlarged and he was admitted to our hospital for further examination and treatment. Transbronchial lung biopsy was performed and serologically, bacteriologically and histologically a diagnosis of chronic coccidioidmycosis was made. It is very rare for fungus ball formation and coexistence of spherules and hyphae of Coccidioides immitis to be seen. Fluconazole was temporarily effective, causing cavities to shrink and eosinophilia to decrease, however Amphotericin B needed to be used later. Eosinophilia was closely related to the severity of the disease gravity. PMID- 17087338 TI - [Case of drug-induced lung injury due to Rebamipide]. AB - A 76-year-old man was admitted because of chest discomfort and diffuse pulmonary interstitial shadows. The diagnosis of drug-induced lung injury due to Rebamipide was made based on the transbronchial lung biopsy specimen and Drug Lymphocyte Stimulating Test (DLST) for Rebamipide (Mucosta). Corticosteroid therapy was effective. PMID- 17087339 TI - [Case of rapid progressive interstitial pneumonia associated with primary Sjogren syndrome]. AB - A 72-year-old woman with a dry cough and dyspnea on exertion was admitted to our hospital. A chest radiograph showed reticular opacities and volume loss in both lower lung fields. She was troubled with xerostomia and her laboratory test showed positive reaction for anti SS-A and SS-B antibody. Labial biopsy led to a diagnosis of primary Sjogren's syndrome (pSjS). Lung biopsy specimens obtained by video-assisted thoracoscopic surgery (VATS) revealed interstitial pneumonia. On the sixth postoperative day, hypoxemia acutely worsened and her chest radiograph showed widespread diffuse ground-glass attenuation. A diagnosis of acute exacerbation was made, and steroid and immunosuppressive therapy was started. In spite of intensive therapy, she died due to respiratory failure. We report a rare case of interstitial pneumonia with pSjS resulting in acute exacerbation. PMID- 17087340 TI - [Case of lung edema occurring as a result of inhalation of fumes from a Teflon coated flying pan overheated for 4 hours]. AB - The patient was a 59-year-old man who had been treated with continuous positive airway pressure for a diagnosis of obstructive sleep apnea syndrome. While cooking buckwheat noodles, the patient fell asleep with a frying pan on the fire. After four hours, he was awakened by the smell of burning as well as pharyngalgia, and visited this hospital. He was found to have hypoxia, and diagnostic imaging showed diffuse concentration increase with peripheral clear zones in bilateral lung fields, a typical finding of pulmonary edema. In view of the peculiar temporal course of his disease and inhalation of Teflon fumes immediately before its occurrence, he was considered to have polymer fume fever with non-cardiogenic pulmonary edema. After admission, he was treated with oxygen inhalation and diuretics, and experienced a rapid improvement of his general condition and findings on diagnostic imaging. In European countries and the United States, cases with a history of inhalation of Teflon fumes have been given a diagnosis of polymer fume fever. This case is of interest as it is apparently the first such disease reported in Japan. PMID- 17087341 TI - [Case of acute respiratory distress syndrome in which high-frequency oscillatory ventilation (HFOV) was effective for the management of severe hypoxemia]. AB - A 69-year-old man was admitted to our hospital because of acute prostatitis. After admission he produced bloody sputum and chest radiograph showed bilateral infiltration of the upper lung fields. The fraction of neutrophils in the bronchoalveolar lavage fluid was 15.6%, lymphocytes were 33.6%, eosinophils were 26.6% and macrophages were 23.6%. No special findings were observed in the lung tissue obtained by transbronchial lung biopsy. After admission, he displayed acute respiratory distress syndrome with severe hypoxemia as dyspnea gradually progressed. We commenced treatment with high-dose pulse methylprednisolone and neutrophil elastase inhibitor under mechanical ventilation. Because severe hypoxemia was unresolved, HFOV was initiated on the 8th hospital day. His respiratory condition rapidly improved during HFOV. On the 17th hospital day, he was weaned from mechanical ventilation. HFOV seem to be safe and effective for patients with severe ARDS who failed conventional ventilation support. PMID- 17087342 TI - [Radiological findings in initial pulmonary alveolar proteinosis detected in the post-treatment course of nocardiosis]. AB - Chest CT detected a small localized ground glass opacity in the right upper lung in a 52-year-old woman being treated for nocardiosis. A PAS-stain positive material and elevated surfactant level were confirmed in bronchoalveolar lavage fluid, then a diagnosis of pulmonary alveolar proteinosis was established. In early pulmonary alveolar proteinosis with focal opacity, HRCT can demonstrate the substantial findings of alveolar proteinosis such as a crazy-paving appearance or geographic distribution. We should note that alveolar proteinosis in the early stage is easily overlooked and, in addition, nocardiosis might overlap with alveolar proteinosis. PMID- 17087343 TI - [Case of eosinophilic bronchitis and bronchiolitis associated with increased level of serum CEA in asthmatics]. AB - A 50-year-old asthmatic woman showed peripheral blood eosinophilia, significantly increased level of CEA in serum (102.5ng/ml), and atelectasis of the right middle lobe on chest radiograph and CT. The level of CEA subsequently increased further, and then decreased with systemic corticosteroid therapy. Pathological findings of surgically biopsied lung showed eosinophilic broncho-bronchiolitis, without malignant cells. Two years later, chest CT demonstrated various findings such as mucoid impaction, peripheral bronchiectasis and centrilobular nodules, but allergic bronchopulmonary mycosis was not proved. These findings suggest that the eosinophilic bronchiolar inflammation in bronchial asthma caused an increase in serum CEA levels and various findings on chest CT. PMID- 17087344 TI - [Case of Heerfordt's syndrome with prolonged peripheral nerve involvement]. AB - A 28-year-old man complaining of myiodesopsia was given a diagnosis of uveitis. Subsequently he complained facial nerve palsy and enlargement of parotid gland. Heerfordt's syndrome was diagnosed based on the results of several examinations. Facial nerve palsy, enlargement of the parotid gland and uveitis were improved by systemic corticosteroid therapy. At present he is receiving systemic corticosteroid therapy, but numbness in the mouth, thought to be the involvement of the trigeminal nerve, remains. Systemic corticosteroid therapy is usually effective for most cases with Heerford's syndrome. On the other hand, there are some cases with the prolonged peripheral nerve involvement despite systemic corticosteroid therapy, as seen in this case. If peripheral nerve involvement is prolonged, it is necessary to consider small-fibre neuropathy as one possible cause. PMID- 17087345 TI - [Successful treatment by voliconazole for pulmonary and adductor magnus muscle aspergillosis induced by immunosuppressive therapy for hypersensitivity pneumonia]. AB - A 74-year-old woman was treated with steroid and cyclosporine A for hypersensitivity pneumonia. To examine the causes of general fatigue and increased levels of beta-D glucan in serum, she was admitted to our hospital. Chest computed tomography (CT) scan revealed nodular opacity with a well-defined margin in the right S1. 67Ga scintigraphy image showed high uptake in the left thigh and CT showed circularly enhanced lesions in the thigh. An ultrasonography guided needle aspiration and biopsy of the muscle abscess allowed isolation of Aspergillus fumigatus and evidence of necrotic tissues around the granuloma formation. We therefore diagnosed invasive aspergillosis. Because of the poor response to initial therapy with micafungin and itraconazole for 4 weeks, we treated her with voliconazole (VCZ). Spectacular regression of lung lesions and muscle abscesses was rapidly achieved. Furthermore, the high level of beta-D glucan in serum decreased gradually. This case suggests that administration of VCZ can be recommended for deep seated mycoses. PMID- 17087346 TI - [Case of obstructive sleep apnea syndrome exacerbated due to cervical spinal cord injury]. AB - A 70-year-old man injured his spinal cord at the fourth cervical vertebra due to a fall and became quadriplegic. Following emergency treatment, the patient was admitted to our hospital for rehabilitation. On admission, respiratory function tests indicated a percent vital capacity value (%VC) of 58.6%, revealing restrictive ventilatory impairment. After admission, we observed difficulty breathing at night and extreme snoring and thus performed a simple full-night polysomnography. Results showed an apnea-hypopnea index (AHI) of 46.5/hr and serious obstructive sleep apnea syndrome (OSAS). Based on these results, we conjectured that a preexisting OSAS condition was exacerbated by the cervical spinal cord injury, and thus administered nasal continuous positive airway pressure (NCPAP) in addition to respiratory rehabilitation. A speedy disappearance of clinical symptoms was found with the application of NCPAP. Compared to a 3% oxygen desaturation index (ODI 3%) of 31.0 dips/hr prior to treatment, the ODI 3% dropped to 3.5 dips/hr after 14 days, and 0 dips/hr after 60 days, indicating an obvious improvement in nocturnal oxygen desaturation. PMID- 17087347 TI - [Case of drug-induced pneumonitis associated with a dietary supplement containing CoQ10]. AB - A 61-year-old woman began to take a dietary supplement contained CoQ10 and perilla leaf extract. Two months later, a dry cough appeared. The number of eosinophils in peripheral blood was elevated and chest radiograph images showed infiltrative shadows in the left middle lung. A chest CT scan showed consolidation in the left upper lobe (S3) and lower lobe (S10). The percentage of eosinophils was abnormally high in bronchoalveolar lavage fluid (BALF), and examination of a transbronchial lung biopsy (TBLB) specimen showed marked infiltration of eosinophils in the alveolar wall. Drug lymphocyte stimulation test (DLST) indicated high stimulation index for both supplement containing CoQ10 and its element of CoQ10. We diagnosed drug-induced pneumonitis, caused by CoQ10. The symptoms of the patient and pulmonary infiltrative shadows on chest radiograph improved after she stopped taking the supplements and started taking prednisolone orally. Recently various dietary supplements are coming onto the market. Since the possible adverse effects of these supplements are not investigated extensively, care should be taken for symptoms relating to food supplements. PMID- 17087348 TI - [Case of multiple thin-walled cavitary pulmonary metastasis from bladder cancer]. AB - A 71-year-old man was admitted to the urological section of our hospital because of hematuria. Cystoscopy revealed a bladder tumor. Transurethral resection of the bladder tumor (TUR-Bt) was performed. This tumor was diagnosed pathologically as bladder cancer (transitional cell carcinoma pT2). His chest computed tomogram showed multiple thin-walled cavitary nodules in both lungs. The pathological images of the specimen that was resected by video assisted thoracic surgery (VATS) showed transitional cell carcinoma. A diagnosis of pulmonary metastasis from bladder cancer was made. This is a rare case. PMID- 17087349 TI - [Small cell lung cancer with Sjogren's syndrome and Lambert-Eaton myasthenic syndrome]. AB - A 72-year-old man was admitted to our hospital with complaints of dry mouth, muscle weakness of the lower limbs and gait disturbance. The patient had dry mouth, dry eyes, positive anti-SS-B antibody and salivary gland inflammation. Sjogren's syndrome was diagnosed. Since muscle weakness of the lower limbs and gait disturbance were not compatible with Sjogren's syndrome, we considered the possibility of paraneoplastic syndrome. Serum levels of CEA, NSE and ProGRP were elevated. Chest roentgenogram and CT showed a nodular lesion in the right upper lobe of the lung and swollen lymph nodes in the hilum and mediastinum. Small cell lung cancer was diagnosed by bronchoscopy. Anti-P/Q-type Ca2+ channel antibody was positive. Electromyogram showed a reduction in the amplitude of the evoked muscle action potential response after slow repetitive stimulation and did not show a reduction after rapid repetitive stimulation. Based on these findings, we made a diagnosis of Lambert-Eaton myasthenic syndrome (LEMS). Concurrent chemoradiotherapy induced an improvement of muscle weakness of the lower limbs. LEMS is frequently associated with a malignant tumor and an autoimmune disorder. We thought that in this patient, the presentation of LEMS was apparent because he had both Sjogren's syndrome and small cell lung cancer. PMID- 17087350 TI - [Case of primary cavitary sarcoidosis: exacerbation after 16 years from onset]. AB - A 46-year-old man presented with the complaint of cough, fever and dyspnea on exercise. Sixteen years previously stage II sarcoidosis was diagnosed, with an ocular lesion. So far, he had had no respiratory symptoms and was followed up without medication. Two months before his visit, he had a cough. He gradually developed a fever and felt dyspnea on exercise. Chest X-ray showed an infiltrative shadow with a cavity in the right upper lobe. Bronchofiberscopy was performed and the biopsy specimen from the cavity wall revealed noncaseating epitheloid cell granulomas, suggesting primary cavitary sarcoidosis. Corticosterid therapy was started. His symptoms improved rapidly, and the infiltrative shadow on the right upper lobe decreased within one month. The cavity wall was thin on the following computed tomography. Thirty-two cases of primary cavity sarcoidosis were reported in Japan from 1975-2005. The mean age of the patients was 26.5 years old and the ratio of males to females was 13:3. In most cases, corticosteroid therapy was effective and nine spontaneously improved cases were reported. To determine the indications for steroid therapy, further examination is needed. PMID- 17087351 TI - [Roles and molecular mechanisms of chromosome cohesion at centromeres]. PMID- 17087352 TI - [Visual system of Drosophila melanogaster: a model system for understanding mechanisms underlying brain development]. PMID- 17087353 TI - [Human transcription factors as typical examples of those proteins with intrinsically disordered regions]. PMID- 17087354 TI - [Progress in research on vitamin D]. PMID- 17087355 TI - [Circannual control of hibernation by hibernation-specific protein (HP) complex in mammals]. PMID- 17087356 TI - [Generation of neural cells from embryonic stem cells]. PMID- 17087357 TI - [Japanese biochemistry researchers in Europe]. PMID- 17087358 TI - [Transdirect insect cells]. PMID- 17087359 TI - [Recent progress in research and development of RNA-based pharmaceuticals]. PMID- 17087360 TI - [Secular trends and current status of lifestyle-related diseases: the Hisayama Study]. PMID- 17087361 TI - [Asia-Pacific telemedicine project via high-speed internet]. PMID- 17087362 TI - Both hepatitis C virus and Chlamydia pneumoniae infection are related to the progression of carotid atherosclerosis in patients undergoing lipid lowering therapy. AB - Recent experimental and epidemiological findings suggest that infectious agents may play a role in the development and progression of atherosclerosis. We previously reported that Chlamydia pneumoniae (C. pneumoniae) infection reduces the effectiveness of lipid-lowering therapy for carotid atherosclerosis and that this micro-organism may play a role in the progression of atherosclerosis. In this study, we investigated the possible association between hepatitis C virus (HCV) infection and carotid arteriosclerosis. A total of 165 asymptomatic hypercholesterolemic patients were randomized to receive probucol (500 mg/day, n=82) or pravastatin (10 mg/day, n=83) and were followed for 2 years. The 2-year change of the maximum common carotid artery intima-media thickness (Max-IMT) was the primary endpoint, while the Max-IMT and the incidence of major cardiovascular events were secondary endpoint. All serum samples were tested for antibody to HCV (anti-HCV) by enzyme-linked immunosorbent assay (ELISA), and all anti-HCV positive samples were assayed for HCV RNA. Patients without HCV infection (n=25) showed a significant reduction of Max-IMT (-10.9%) (p<0.0001), while a small decrease of Max-IMT was noted in the patients with HCV infection (n=25) (-0. 3%). Significant differences in the reduction of serum total cholesterol and LDL cholesterol were found between patients with and without HCV infection (both p<0.0001). No significant difference in therapeutic effect was noted between the probucol and the pravastatin groups. After adjustment for confounding risk factors, both C. pneumoniae infection and anti-HCV positivity were associated with a greater risk of an increase in Max-IMT (8.5635 [1.3738-15.7532], p<0.05, 9.5040 [0.2886-18.7194], p<0.05, respectively). These findings suggest that both chronic HCV infection and C. pneumoniae infection can reduce the effectiveness of lipid-lowering therapy for carotid atherosclerosis, and that the HCV may play a role in the progression of atherosclerosis in HCV infected patients. PMID- 17087363 TI - European developments in standardisation of sludge physical parameters. AB - Characterisation is an important step in sludge management as it allows sludge properties to be evaluated and behaviour predicted. This is well recognised by the European Union (EU) countries which consider necessary the development of standardised sludge characterisation methods and procedures because objective and transparent regulations allow sludge management to be properly performed, legal requirements correctly fulfilled, and public confidence built. To this end, the European Committee for Standardization (CEN) established the Technical Committee 308 (TC308) whose scope is the production of Standards for sludge characterisation, and of Guides of good practice. In this field, physical properties are of great importance as they allow the prediction of sludge behaviour when handled and submitted to almost all treatment and utilisation/disposal operations. Activity of CEN/TC308 is developed in three Working Groups (WG) and several Task Groups (TG); in particular, TG3 of WG1 deals with physical parameters. In this paper developments regarding Standards for evaluation of capillary suction time, specific resistance to filtration, compressibility, settleability, thickenability, and calorific value, and Technical Reports dealing with a procedure for laboratory chemical conditioning and with sludge consistency (flowability/solidity) are briefly outlined. The results of relevant inter-laboratory tests for the validation of above standards are summarised. PMID- 17087364 TI - Development of the South African wastewater sludge guidelines. AB - The new South African Wastewater Sludge Guideline Series will replace the document entitled: "Permissible Utilisation and Disposal of Sewage Sludge, Edition 1, 1997" which deals with application of sewage sludge to land. The new guideline series will be published as 5 volumes: Volume 1: Management Options for the Use or Disposal of Wastewater Sludge, Volume 2: Requirements for the agricultural use of wastewater sludge, Volume 3: Requirements for the on site and off-site disposal of wastewater sludge, Volume 4: Requirements for the use of wastewater sludge in the production of commercial products, Volume 5: Requirements for the incineration of wastewater sludge, We envisage that Volumes 1 and 2 will be published at the time of the conference and that work has begun on Volume 3, 4 and 5. The paper aims to communicate the major differences between the previous sludge guidelines and these new sludge guideline series. For example, the paper details the new wastewater sludge classification system and the requirements for the characterisation of the sludge for classification purposes. The paper will also explain the approach followed in the development of Volume 1 and 2 and will elaborate on the proposed approach for Volumes 3, 4 and 5. PMID- 17087365 TI - Toward efficient sludge processing using novel rheological parameters: dynamic rheological testing. AB - Materials known as complex fluids exhibit a transitional behaviour between their solid and liquid states. Sludges produced at different treatment stages of wastewater treatment are examples of these materials, since they exhibit solid behaviour within shorter time frames and liquid behaviour over longer durations. These sludges can also change from solid-like to liquid-like, or vice versa, when subjected to even a slight deformation; this stems from their complex makeup, which includes both particulate materials and polymeric gels, including both biopolymers and the synthetic polymers used in conditioning them. This paper focuses on two means of quantifying dynamic rheological test results: (1) the linear viscoelastic behaviour (LVE) of sludge samples, as measured by the strain sweep analysis storage (G') and loss moduli (G"); and (2) the flow curve, which presents the relationship between the shear viscosity and shear rate and determines the yield stress values of samples using the strain sweep test mode. With these methods, sludges can be more accurately characterised, allowing better prediction of their behaviour in treatment and transport processes. PMID- 17087366 TI - Evaluation of conditioning responses of thermophilic-mesophilic anaerobically and mesophilic aerobically digested biosolids using rheological properties. AB - One of the most crucial processes in biosolids management is stabilisation, which is typically accomplished using either anaerobic or aerobic digestion processes. Although there are many advantages to both of these methods--such as reduction of volatile organic matter, pathogen content, and offensive odours--one common disadvantage is that digested biosolids exhibit poor dewaterability characteristics. The deterioration in dewaterability of digested biosolids leads to increases in polymer requirements, connoting higher conditioning costs. Many studies have examined this phenomenon based on conventional filterability tests such as capillary suction time and specific resistance to filtration. However, these test methods are limited in their ability to predict full-scale dewatering behaviour (particularly in centrifugation), and do not regard handling properties as being important for sludge transport. For this purpose, new rheometric methods may assist in predicting sludge behaviour, potentially allowing optimisation of stabilisation and conditioning processes relative to the desired sludge properties. This paper investigates the possible utility of these methods. Classical methods for characterising biosolids, such as filtration tests, are compared with yield stress values and other rheological properties as determined using the steady-shear rate sweep test. The rheological behaviours are compared for different sludge types, including raw and polymer conditioned thermophilic anaerobically, mesophilic anaerobically, and mesophilic aerobically digested biosolids. The paper demonstrates the determination of specific energy values associated with breakdown of the flocculated matrix. Robust modelling approaches are employed to optimise the conditioning of the digested samples, and to shed light on commonly used rheological models such as Bingham, Herschel-Bulkley, and Ostwald equations. PMID- 17087367 TI - Enhanced sludge conditioning by enzyme pre-treatment: comparison of laboratory and pilot scale dewatering results. AB - The effect of enzyme pre-treatment on dewaterability of anaerobically digested sludge was investigated at both laboratory and pilot scale. Our results revealed a significant increase in cake solid content (27% cake solids compared to 18% without enzyme pre-treatment), using an enzyme dose of only 20 mg/L. In order to assess practical application, enzyme pre-treatment was applied at the Wilmington, Delaware (U.S.) wastewater treatment plant, using a pilot-scale centrifuge. However, the efficiency reached in laboratory scale could not be obtained in pilot scale, where the final cake solids content did not exceed 20%. Centrifuge and belt filter press (simulated by Crown Press) dewatering were compared in terms of the process efficiencies in the absence and presence of enzyme pre treatment. Possible factors that might cause the differences were tested by experimental and statistical comparisons. Results indicated that the higher shear applied in centrifugation is responsible for the lack of improved cake solids. The network strength of sludge determined by rheological measurements revealed that enzymatic treatment weakens the gel structure of the sludge floc through the hydrolysis of extracellular polymeric substances; this allows improved dewatering by filtration processes, but leads to floc deterioration when subjected to high shear during centrifugation. PMID- 17087368 TI - Influence of anaerobic digestion on particle surface charge and optimal polymer dosage. AB - Anaerobic digestion leads to significant changes of the sludge structural matrix, affecting particle size distribution and dewaterability. The surface charge, determined by means of streaming current, can be effectively used to monitor the complex phenomena of floc disruption, colloid formation and chemical conditioning. To study the relation between surface charge and optimal dosage, two different cationic polyelectrolytes were used: Praestol 644, polymer with high molecular weight and low charge density, and Poly Dadmac, with relatively low molecular weight but high charge density. The optimal Poly Dadmac dosage strictly met the value required to neutralise particle charge whereas the optimal dosage of Praestol 644 indicated that the relevant charge was considerably lower than the one required for charge neutralisation. Mechanisms of action are therefore clearly different. Another objective was to investigate the changes of dewatering characteristics of secondary sludge during anaerobic digestion tests at different inoculum content by determining charge density, and optimal polymer dosage. The optimal polyelectrolyte dosage remains almost constant during digestion at high inoculum, but a significant increase in the first period is observed at low inoculum, thus suggesting that the release of colloidal and supracolloidal material from sludge affects dewaterability, especially in the first days of digestion. PMID- 17087369 TI - Biological sludge solubilisation for reduction of excess sludge production in wastewater treatment process. AB - A novel sludge disintegration system (JFE-SD system) was developed for the reduction of excess sludge production in wastewater treatment plants. Chemical and biological treatments were applied to disintegrate excess sludge. At the first step, to enhance biological disintegration, the sludge was pretreated with alkali. At the second step, the sludge was disintegrated by biological treatment. Many kinds of sludge degrading microorganisms integrated the sludge. The efficiency of the new sludge disintegration system was confirmed in a full-scale experiment. The JFE-SD system reduced excess sludge production by approximately 50% during the experimental period. The quality of effluent was kept at quite a good level. Economic analysis revealed that this system could significantly decrease the excess sludge treatment cost. PMID- 17087370 TI - Organic matter release in low temperature thermal treatment of biological sludge for reduction of excess sludge production. AB - Thermal treatment applied in association with a biological system allows for a significant reduction in excess sludge production (approximately 50%). In general, heat treatment is described as a sludge disintegration technique. This paper offers a thorough study on the impact of heat treatment, at temperatures below 100 degrees C, on the solubilisation of the sludge COD and its biodegradability. Discontinuous heating experiments were performed on activated and digested sludge. At all temperatures tested the released COD for digested sludge was systematically higher than that for activated sludge (15 and 40%, respectively, at 95 degrees C for 40 min of contact time). For the first 30 min, a 1st order kinetic, with respect to the residual COD, was systematically found. In the range of 40-95 degrees C, digested sludge had a lower activation energy than activated sludge (26 kcal/mol compared to 70-160 kcal/mol). COD solubilisation is thus more positively influenced by temperature in the case of activated sludge. This may be due to the significant difference in the ratio of protein/carbohydrate in digested and activated sludge (1-5 and 0.2-0.7, respectively). The increase in the COD/TKN ratio in the solubilised fraction after thermal treatment of activated sludge suggests a preferential solubilisation of proteins over carbohydrates. Respirometric tests performed on the solubilised COD showed that whatever the sludge origin, only 40-50% of released COD is biodegradable at a conventional hydraulic retention time (i.e., 24 h). Hence, heat treatment would act more through organic matter solubilisation rather than by a biodegradability increase. PMID- 17087371 TI - Reduction of excess sludge production using mechanical disintegration devices. AB - The usability of mechanical disintegration techniques for the reduction of excess sludge production in the activated sludge process was investigated. Using three different disintegration devices (ultrasonic homogeniser, stirred media mill, high pressure homogeniser) and different operational parameters of the disintegration, the effect of mechanical disintegration on the excess sludge production and on the effluent quality was studied within a continuously operated, laboratory scale wastewater treatment system with pre-denitrification. Depending on the operational conditions and the disintegration device used, a reduction of excess sludge production of up to 70% was achieved. A combination of mechanical disintegration with a membrane bioreactor process with high sludge age is more energy effective concerning reduction of sludge production than with a conventional activated sludge process at lower sludge ages. Depending on the disintegration parameters, the disintegration has no, or only minor, negative effect on the soluble effluent COD and on the COD-removal capacity of the activated sludge process. Nitrogen-removal was slightly deteriorated by the disintegration, whereas the system used was not optimised for nitrogen removal before disintegration was implemented. PMID- 17087372 TI - Effect of feed/inoculum ratio on anaerobic digestion of sonicated sludge. AB - In recent years, relevant interest has been devoted to activated sludge disintegration and solubilisation techniques in order to cope with the biological limitations related to particulate degradation. Mechanical disintegration with ultrasound can efficiently transform insoluble organics into a soluble form: the solubilised organic matter is released from the cells to the bulk phase, thus accelerating the hydrolysis step in the digestion process. Experiments were carried out on bench scale anaerobic reactors fed with either untreated or disintegrated excess sludge, added with a biomass inoculum taken from a full scale anaerobic digester. Digestion tests have been carried out at different feed/inoculum ratios (F/I) in the range of 0.1-2, kinetics of VS reduction has been investigated and a beneficial effect of sonication is observed for all the experimental conditions. Similar beneficial results have also been found for biogas production with a maximum gain of 25% at 0.5 F/I ratio. PMID- 17087373 TI - The feasibility study of using non-woven MBR for reduction of hydrolysed biosolids. AB - In this study, non-woven MBR was used to treat hydrolysed biosolids wasted from a biological treatment plant. The concentration of SS of hydrolysed biosolids in influent was 10,000 mg/L and the concentration of SS in effluent was less than 200 mg/L with/without discharging wasted sludge depending on different HRTs, i.e. 20, 15 and 10 d. The results indicated that the percentage of biosolids reduction in terms of SS removal efficiency in non-woven MBR was around 65, 60 and 35%, respectively, depending on different HRTs. Meanwhile, the ratio of VSS/SS was decreased from 0.78 to 0.50 and the number of smaller inorganic particle sizes increased due to extended SRT. The initial flux in the non-woven MBR was set at 0.02, 0.04 and 0.06 m3/m2/day and trans-membrane pressure (TMP) was less than 10 kPa. The permeate flux could be maintained quite stably due to lower TMP. The proposed non-woven MBR could be used to achieve the reduction of biosolids in the wastewater treatment plant. PMID- 17087374 TI - The effects of hydraulic retention time and feed COD concentration on the rate of hydrolysis of primary sewage sludge under methanogenic conditions. AB - A series of completely mixed methanogenic anaerobic digesters have been operated to determine the rate of hydrolysis of primary sewage sludge. The hydraulic retention time was reduced from 60 d to when the system failed (approximately 5 d), while the feed COD concentration was 40, 25, 13 and 2 gCOD/L. A steady state model based on first order kinetics was developed to simulate the hydrolysis rate at each retention time and feed concentration. With the mean value for the hydrolysis rate constant (0.992 +/- 0.492 d(-1)), this model was able to accurately predict the effluent COD for all steady state operating conditions. However, the effluent COD concentration was relatively insensitive to the exact value for this constant. The model provides a framework for analysis of anaerobic digestion experimental data, to enable meaningful comparisons. PMID- 17087375 TI - Maximising biogas in anaerobic digestion by using engine waste heat for thermal hydrolysis pre-treatment of sludge. AB - Dublin's Ringsend WWTP was designed to serve a population of approximately 1.2 million p.e. with a sludge production of 37,000 dry tonnes per year after upgrading to full secondary treatment. Several technical solutions were put forward as part of a design, build, finance and operate (DBFO) competition, with the chosen solution being a proposal by Black and Veatch for a combination of sequencing batch reactor (SBR) technology and anaerobic digestion with Cambi thermal hydrolysis pre-treatment (THP). The THP plant was built by Cambi and handed over to B&V in 2002. The plant is now operated by Celtic Anglian Water. In September 2004 a test was carried out on the mass and energy balance of the plant following 2 years of operation and is detailed in this paper. The process enables digestion at very high dry solids feed and low hydraulic retention time. The plant was built with three digesters of 4250 m3 each and is fed with hydrolysed sludge at 11% DS. There are four no. 1 MW Jenbacher engines operating mainly on biogas. Each pair of engines is fitted with a waste heat boiler with a capacity of one tonne steam per hour. These boilers have sufficient capacity to provide 80% of the steam required for the THP, which in turn provides all the heat for the subsequent digestion in the form of hydrolysed feed. There are two main biogas boilers for top up steam and other uses of the biogas including thermal oxidation of concentrated odours. PMID- 17087376 TI - Integrated chemical, physical and biological processes modelling of anaerobic digestion of sewage sludge. AB - The biological kinetic processes for anaerobic digestion (AD) are integrated into a two phase subset of a three phase mixed weak acid/base chemistry kinetic model. The approach of characterising sewage sludge into carbohydrates, lipids and proteins, as is done in the International Water Association (IWA) AD model No 1 (ADM1), requires measurements that are not routinely available on sewage sludges. Instead, the sewage sludge is characterised with the COD, carbon, hydrogen, oxygen and nitrogen (CHON) composition and is formulated in mole units, based on conservation of C, N, O, H and COD. The model is calibrated and validated with data from laboratory mesophilic anaerobic digesters operating from 7 to 20 d sludge age and fed a sewage primary and humus sludge mixture. These digesters yielded COD mass balances between 107-109% and N mass balances between 91-99%, and hence the experimental data is accepted as reasonable. The sewage sludge COD is found to be 32-36% unbiodegradable (depending on the kinetic formulation selected for the hydrolysis process) and to have a C3.5H7O2N0.196 composition. For the selected hydrolysis kinetics of surface mediated reaction (Contois), with a single set of kinetic and stoichiometric constants, for all retention times good correlation is obtained between predicted and measured results for: (i) COD; (ii) free and saline ammonia (FSA); (iii) short chain fatty acids (SCFA); (iv) H2CO3 * alkalinity; (v) pH of the effluent stream; (vi) CO2; and (vii) CH4 gases in the gas stream. The measured composition of primary sludge from two local wastewater treatment plants ranged between C3.38H7O1.91 N0.21 and C3.91H7O2.04N0.16. The predicted composition based on mass balances is therefore within 5% of the average measured composition providing persuasive validation of the model. PMID- 17087377 TI - Extraction efficiency of phosphate from pre-coagulated sludge with NaHS. AB - The recovery of phosphorus from sewage and sludge treatment systems is particularly important because it is a limited resource and a large proportion of the phosphorus currently used in Japan must be imported. We have been experimentally evaluating recovery methods with sulphide. In this study, we focussed on the extraction of phosphate from the sludge, and sought to achieve a greater extraction efficiency and to validate the extraction mechanism. We conducted three experiments, i.e. a sludge-type experiment, a coagulant ratio of pre-coagulated sludge experiment, and a concentration of pre-coagulated sludge experiment. Phosphate was extracted not with normal sewage sludge but with pre coagulated sludge and FePO4 reagent at S/Fe = 1.0-2.0. A coagulant ratio of 23mg Fe L(-1) was required in the precoagulation process to effectively extract phosphate. A high concentration of pre-coagulated sludge was required for the phosphate extraction. The mass balance was calculated, and 44.0% of phosphorus was extracted to supernatant, and 98.5% of iron and 98.3% of sulphur (44.1% of sulphur was sulphide). Thus, phosphate can be selectively separated from iron by the phosphate extraction method with NaHS, and phosphorus and iron can be recovered and reused at sewage treatment plants using ferric chloride as a coagulant. PMID- 17087378 TI - Agricultural use of municipal wastewater sludges: phosphorus availability of biological excess phosphorus removal sludges. AB - Reuse of sewage sludges as phosphorus fertiliser requires the estimation of the plant availabilities of phosphorus (P) from different sludges. This study investigates the effect of lime stabilisation on the phosphorus availability from biological phosphorus removal sludges. In the first part of the study, pot experiments were carried out to assess the fertilising effect of a dewatered biological phosphorus removal sludge. Availability of P was determined in terms of plant-uptake. In the second part of the study, incubation tests were carried out to observe the change in the available P with time when the waste activated sludge (WAS) from an enhanced biological phosphorus removal (EBPR) process is mixed with the same soil. In this part, the plant available P was measured in terms of Olsen extractable P. A P-deficient, alkaline soil was used in the experiments and Lollium Perenne was selected as the testing plant. The results of the pot experiments revealed that lime-stabilisation of the sludge considerably decreased or retarded the availability of P in the sludge. In the incubation tests, the availability of phosphorus in the lime stabilised and nonstabilised sludge amended soil samples was close to each other. In general, P-availability was increased due to the sludge application except for the lime-stabilised dewatered sludge. PMID- 17087379 TI - Dedicated land disposal of wastewater sludge in south africa: leaching of trace elements and nutrients. AB - Most of the wastewater treatment facilities in South Africa (80%) dispose of their sewage sludge on dedicated land disposal (DLD) sites. The impact of this practice on the environment is believed to be negative, but very little research has been carried out to determine the extent of the damage to the soil and water resources. Forty wastewater treatment facilities using DLD, with different soil properties, application techniques, metal concentrations and period of sludge application, were studied. Soil and groundwater samples were collected at each of the selected facilities. Three extraction methods (aqua regia, NH4EDTA and NH4NO3) were used and samples were analysed for total N, P and K, pH, organic carbon and their metal content (Cr, Co, Ni, Cu, Zn, Cd, and Pb). Some degree of leaching of the heavy metals (especially Co and Ni) occurred at some of the sampling sites and the average depth of leaching was 100-200 mm. Deeper than 300 mm, the metal concentrations in most soil samples reached background concentrations. Seven of the nine groundwater samples that could be obtained had high NO3 concentrations (> 6 mg L(-1)). Statistical analyses of the data indicate no significant differences between sludge type (wet or dry) and leaching, or age of the disposal sites and leaching. Taking into account the age of the disposal sites, the frequency of sludge application and the metal load of the sludge, the depth of leaching is surprisingly shallow in most soils, in spite of the low soil pH(H2O) and clay content. PMID- 17087380 TI - Heavy metal displacement in EDTA-assisted phytoremediation of biosolids soil. AB - Chelating agents are added to soil to solubilise the metals for enhanced phytoextraction. Yet no studies follow the displacement and leaching of heavy metals in soil with biosolids following solubilisation with chelates. The objective of this work was to determine the mobility of heavy metals in biosolids in a soil from a sludge farm that had received biosolids for 25 years. The soil was placed in columns in a greenhouse. Columns either had a plant (poplar) or no plant. After the poplar seedlings had grown for 144 d, the tetrasodium salt of the chelating agent EDTA was irrigated onto the surface of the soil at a rate of 1 g per kg of soil. Drainage water was analysed for three toxic heavy metals and four essential heavy metals. Without EDTA, concentrations of the seven heavy metals in the leachate from columns with or without plants were low or below detection limits. With or without plants, the EDTA mobilised all heavy metals and increased their concentration in drainage water. Without plants, the concentrations of Cd, Cu, Fe, and Zn in the leachate from columns with EDTA were above drinking-water standards. The presence of poplar plants in the soil reduced the concentrations of Cu, Fe, and Zn in the leachate so it fell within drinking water standards. Concentrations of Cd and Pb in the leachate remained above drinking-water standards with or without plants. PMID- 17087381 TI - An investigation of agricultural use potential of wastewater sludges in Turkey- case of heavy metals. AB - Landfilling is one of the most common methods that are used in the final disposal of sludge world wide, as well as in Turkey. However, increasing sludge quantities and limited landfilling areas make this method difficult to apply. The use of sludge in agriculture presents a possible alternative for disposal. However, it also poses some risks to be evaluated. In this respect, it is important to identify the heavy metal content of sewage sludges because of its adverse health effects. This study aims to determine the heavy metal contents of sludges and wastewaters from three different wastewater treatment plants in Turkey. The selection of plants was carried out according to the different treatment technologies applied to wastewater and sludge in those plants. Heavy metal analysis of sludges and wastewaters was conducted using a microwave assisted digestion procedure. After sampling and analysis, the results show that all the related metal concentrations are below the values that are set in the Soil Pollution Control Regulation of Turkey. However, in two of the three plants, Zn and Ni concentration should be tracked carefully. PMID- 17087382 TI - The survival of pathogens in soil treated with wastewater sludge and in potatoes grown in such soil. AB - The prevalence of pathogens on potatoes (Solanum tuberosum) grown in soil amended with a pathogen rich wastewater sludge was investigated. Bacteria of the family Enterobacteriaceae are important pathogens causing intestinal and systemic illness of humans and other animals. Type B sludge was used. Sludges investigated are the high metal and the low metal sludges. Microorganisms in the sludge amended soil were using culture-based technique. Salmonella and E. coli were observed in tested soil samples. No microorganisms were isolated from control samples taken throughout the process of the experiment. At harvest time, some of the potato samples from LMS soil were contaminated. These potatoes were subjected to further investigation using molecular techniques (polymerase chain reaction) with fD1 and rP2 as primers. Organisms identified from the sequenced potato peel samples with the BLAST search tool included Enterobacter agglomerans (Pantoea agglomerans), several Buttiauxella spp., Pectobacterium spp., Erwinia spp. and a few Pantoea spp. Other than the E. agglomerans, which is commonly found in the gut and upper respiratory tract of humans and in the environment, all the other species identified were found to be mainly either plant or soil pathogens. The E. agglomerans are not primary pathogens but secondary opportunistic pathogens particularly in immunocompromised individuals. These results suggest that growing high risk crops using wastewater sludge contaminated soil may lead to limited infestation of produce with primary pathogens. It appears that the use of HMS due to early pathogen die-off provides less risk of infection than the LMS. However, proper treatment of wastewater sludge to reduce pathogen load is essential prior to its use as soil conditioner. PMID- 17087383 TI - Using Ecosan sludge for crop production. AB - This paper presents the microbial risks associated with the reuse of Ecosan sludge in agriculture. Sludge from KwaZulu Natal Province in South Africa with a helminth ova content of 30 HO/g TS, faecal coliforms of 10(6) CFU/g TS, faecal streptococci of 10(6) CFU/g TS and Salmonella spp. of 10(5) CFU/g TS was used for this purpose. Applying different rates of sludge to spinach and carrots resulted in an increase in bacterial and helminth ova counts in crops as the quantity of sludge increased. The helminth ova content in crops was always greater in leaves than in stems, with a content varying from 2 to 15 HO/g TS for spinach with sludge application rates of 0-37.5 ton/ha and from 2-8 HO/g TS for carrot crops with sludge application rates varying from 0-35 ton/ha. Nevertheless, viability in crops was 20-25%, reducing the risk of disease. Consequently, the development of an Ecosan sludge revalorisation standard is considered important. Some recommendations are made taking into account the characteristics of the sludge as well as conditions in rural areas. PMID- 17087384 TI - Disinfection of sludge with high pathogenic content using silver and other compounds. AB - A physicochemical sludge with high microbial content (10(2)-10(4) FPU/g TS bacteriophages, 10(6)-10(7) MPN/g TS faecal coliforms, 10(4) MPN/g TS Salmonella spp., 10(4) MPN/g TS Shigella spp., 10(3) MPN/g TS Pseudomonas aeruginosa, 10(2) MPN/g TS Vibrio cholerae, 10(2)-10(3) cysts/g TS Giardia sp., 10(2)-10(4) oocyts/g TS Cryptosporidium sp., 168-215 viable helminth ova/g TS) was disinfected using silver, silver-copper, and silver-copper plus a synergistic agent (SA). Twenty milligrams Ag/g TS inactivated 4.8 log of faecal coliforms in 1 h; however, 40 mg Ag/g TS are needed to reduce helminth ova viability from 84 to 38.4% in the same period of time. Combinations of Ag-Cu (60:600 mg Ag-Cu/g TS) and Ag-SA (60:24 mg Ag-SA/g TS) inactivated 7.8 log of faecal coliforms and around 90% of helminth ova in 60 min. To produce USEPA class A biosolids, 10:100:8 and 5:50:13.3 mg Ag-Cu-SA/gTS are needed. Bacterial regrowth was not observed for all conditions producing <1000 MPN/gTS faecal coliforms, suggesting a residual disinfection effect. Recommended doses to produce class A biosolids inactivated 2-4 log of bacteriophages, 4 log of Salmonella spp., 4 log of Shigella spp., 3 log of Pseudomonas aeruginosa, 2 log of Vibrio cholerae, 87 99.9% of Giardia sp., and 75-99.9% of Cryptosporidium sp. PMID- 17087385 TI - Sludge production and management processes: case study in China. AB - In 2010, the sewage sludge production rate will be 178,500 t dried solids (ds) for Beijing and 294,000 t-ds for Shanghai, respectively. Beijing adopts a centralized system to stabilize 78% of her sludge in three rural Stabilization Centres. Aerated composting technique will be used. Shanghai on the contrary decentralizes the management plan to treat the sludge on site. Diverse treatment trains, such as aerobic/anaerobic digestion, drying, incineration, and composting will be applied. Production rate, treatment plan, and the associated costs, energy consumption, carbon dioxide emission, and risk assessment for heavy metals and pathogens on human health were evaluated in this report for sludges yielded in Beijing and Shanghai, China. PMID- 17087386 TI - Potentially toxic element release by fenton oxidation of sewage sludge. AB - The presence, in sewage sludge, of excess levels of the potentially toxic elements (PTE) copper, zinc, chromium, cadmium, nickel, lead and mercury, could impact on our ability to recycle these residues in the future. Far stricter limits on the levels of PTEs are likely in proposed legislation. A method involving the dosing of Fenton's reagent, a mixture of ferrous iron and hydrogen peroxide, under acidic conditions was evaluated for its potential to reduce metal levels. The [Fe]:[H2O2] (w/w) ratio was found to give a good indication of the percentage copper and zinc elution obtainable. Sites with no iron dosing as part of wastewater treatment required extra iron to be added in order to initiate the Fenton's reaction. A significant reduction, in excess of 70%, of the copper and zinc was eluted from both raw primary and activated sludge solid fractions. Cadmium and nickel could be reduced to below detection limits but elution of mercury, lead and chromium was less than 40%. The iron catalyst concentration was found to be a crucial parameter. This process has the potential to reduce the heavy metal content of the sludge and allow the recycling of sludge to continue in a sustainable manner. PMID- 17087387 TI - Dewatered alum sludge: a potential adsorbent for phosphorus removal. AB - Alum sludge refers to the by-product from the processing of drinking water in water treatment works. In this study, groups of batch experiments were designed to identify the characteristics of dewatered alum sludge for phosphorus adsorption. Air-dried alum sludge (moisture content 10.2%), which was collected from a water treatment works in Dublin, was subjected to artificial P-rich wastewater adsorption tests using KH2PO4 as a model P source. Adsorption behaviours were investigated as a function of amount and particle size of alum sludge, pH of solution and adsorption time. The results have shown that pH plays a major role not only in the adsorption process but also in the adsorption capacity. With regard to adsorption capacity, this study reveals the Langmuir adsorption isotherm being the best fit with experimental data (R2 = 0.98-0.99). The maximum adsorption capacities range from 0.7 to 3.5 mg-P/g when the pH of the synthetic P solution was varied from 9.0 to 4.3, accordingly. The outcome of this study indicated that alum sludge is suitable for use as an adsorbent for removal of phosphate from wastewater. PMID- 17087388 TI - The effects of water treatment residues on soil respiration and microbial community structure. AB - Land disposal of water treatment residue (WTR), a by-product of potable water production, is increasingly being considered a viable alternative to land filling. It is crucial, inter alia, that soil quality is not affected adversely. Six different WTRs obtained from South African water treatment facilities were added to Hutton and Westleigh topsoils at rates of 0, 5, 15 and 25% (m/m) to determine their impacts on soil basal respiration. In general, respiration increased with rate of addition of WTR. The Umgeni, Rand, Midvaal and Faure2 WTRs had the greatest effect on respiration. Increases in respiration became less marked with time. The Hutton topsoil was mixed with the WTRs (at 15% m/m) and, following incubation, the bacterial community structure was examined by denaturing gradient gel electrophoresis. Changes in community structure were detected after 5 d and were attributed to physico-chemical changes arising from WTR amendment. However, dominant bands associated with the Hutton soil were retained and microbial diversity was not adversely affected. The findings of this investigation suggest that WTRs can be applied to the Hutton soil at a rate of at least 15% (300 Mg ha(-1)) without having a detrimental effect on microbial indicators of soil quality. PMID- 17087389 TI - The effects of a polyacrylamide-derived water treatment residue on the hydraulic conductivity, water retention and evaporation of four contrasting South African soils and implications for land disposal. AB - Water treatment residue (WTR), a by-product from the production of potable water, was traditionally disposed of to landfill but there is growing interest in applying this waste to land as an alternative disposal option. Because WTR consists mainly of flocculated fine silt and clay, there is concern that should the residue decompose back to its original constituents, there is an adverse risk for soil water storage and transmission properties of soil. In a laboratory study, four different soil types were amended with a polyacrylamide WTR at rates ranging from 0 to 1280 Mg ha(-1). The treatments were evaluated for changes in water retention, hydraulic conductivity and evaporation. The results showed that WTR decreased bulk density and evaporation and actually increased hydraulic conductivity and water retention, mainly because of the sustained performance of the polymer in binding the silt and clay into gravel-sized aggregates. Such changes were, however, only significant at the 1280 Mg ha(-1) application rate. Therefore, very large amounts of WTR would have to be applied in order to markedly change these inherent physical properties of the soils. In view of this finding, the land treatment of WTR appears possible. PMID- 17087390 TI - The REAL process--a process for recycling sludge from water works. AB - In order to produce drinking water, coagulants--such as aluminium salts--are widely used for precipitation and separation of impurities from raw water. The residual from the process is sludge, which presents a disposal problem. The REAL process is a method for recycling the aluminium from the sludge. In a first step, the aluminium hydroxide is dissolved in sulphuric acid. In a second step, an ultra filtration will separate all suspended matter and large molecules, leaving a concentrate of 15-20% dry solids. The permeate will contain the trivalent aluminium ions together with 30-50% of the organic contaminants. In a third step, by concentrating the permeate in a nano filter, the concentration of aluminium will be high enough to, in a fourth step, be precipitated with potassium sulphate to form a pure crystal: potassium aluminium sulphate. The potassium aluminium sulphate is comparable to standard aluminium sulphate. The process will give a residual in form of a concentrate from the ultra filtration, representing a few per cent of the incoming volume. This paper presents the results from a long time pilot-scale continuous test run at Vasteras water works in Sweden, as well as calculations of costs for full-scale operations. PMID- 17087391 TI - Today will determine tomorrow. PMID- 17087392 TI - Stepping up to meet a great need. PMID- 17087393 TI - Designing a user-friendly web site. PMID- 17087394 TI - An update on present and future considerations of aligners. AB - This report reviews the orthodontic treatment of six different patients who received removable aligners. These cases include correction of deep overbite, open bite, mild-to-moderate crowding, large overjet, cases requiring premolar extractions, the presence of multiple restorations, and cases requiring periodontal-restorative treatments, the improved periodontal status with aligners compared to fixed appliances and the use in teenagers. This report demonstrates that a wide range of cases can be effectively treated, provided the cases are thoroughly reviewed at an early stage process using Invisalgn's ClinCheck software, which will show the specific details of all consecutive appliances prior to any treatment being started so as to determine the biologic and biochemical feasibility of treatment. PMID- 17087395 TI - Orthodontic treatment in the early mixed dentition: is this the optimum time to start care? AB - The age at which children should start orthodontic treatment has been debated amongst orthodontists for many decades. Orthodontists can agree on what is a quality orthodontic result, but disagree as to how and when to best obtain this result Some orthodontists contend that starting treatment in the primary dentition is the most effective means of orthodontic care. Other orthodontists would prefer to begin in the early or late mixed dentition. Still others would rather postpone treatment until the permanent dentition at approximately age 12. This article will evaluate the pros and cons of initiating treatment at different ages. PMID- 17087396 TI - Clinical applications of mini-implants as orthodontic anchorage and the peri implant tissue reaction upon loading. AB - Orthodontic tooth treatment depends on anchorage for improved results. There are many different sources of orthodontic anchorage. Segments of teeth or the entire arch have been the most common type of orthodontic anchorage. But in challenging situations, orthodontists frequently need extra-dental supplements of anchorage such as headgear, face mask, and intermaxillary elastics. Most of them require the patient's compliance. Recently, temporary mini-implants placed within the bone tissue have been used as orthodontic anchorage. It has been proven in many studies and case reports that the mini-implant is a very reliable anchorage source clinically and histologically. The purpose of this article is to introduce the basic clinical application of mini-implants as orthodontic anchorage and to discuss basic concepts about the tissue reaction of peri-implant bone upon placement and loading either from orthodontic mechanics and/or function in the orthodontic treatment of the patients. It is possible for mini-implants to supply absolute anchorage even though they may move slightly within the bone tissue without losing clinical stability. The primary application of mini-implants as orthodontic anchorage will be cases that need absolute anchorage for desired tooth movement. PMID- 17087397 TI - Pacific Craniofacial Team and Cleft Prevention Program. AB - There is no doubt modern genetics have greatly influenced our professional and personal lives during the last decade. Uncovering genetic causes of many medical and dental pathologies is helping to narrow the diagnosis and select a treatment plan that would provide the best outcome. Importantly, having an understanding of multifactorial etiology helps direct our attention toward prevention. We now understand much better our own health problems. In some cases, we can modify our lifestyle and diet in order to prevent "environmental factors" from triggering the mutated genes inherited from our parents. Good examples are diabetes and cardiovascular diseases. If we realize we might have inherited genes for cardiovascular problems from several ancestors who had heart attacks, we already know that these genes will make us only "susceptible" for disease. Those who exercise, watch one's weight, diet, and carefully monitor one's lifestyle will very likely--though possessing "susceptibility genes"--stay healthier and, maybe, will never experience any cardiovascular problems. In principle, the same applies for craniofacial anomalies, especially for nonsyndromic cleft lip and palate. One needs to understand genetic and environmental causes of nonsyndromic orofacial clefts in order to prevent them. With all this in mind, the Pacific Craniofacial Team and Cleft Prevention Program have been established at the Department of Orthodontics, University of the Pacific Arthur A. Dugoni School of Dentistry in San Francisco. A partnership with Rotaplast International, Inc., has made it possible for the faculty, orthodontic residents, and students to participate in 27 multidisciplinary cleft medical missions in underdeveloped and developing countries by donating professional and educational services, and, last but not least, by collecting valuable data and specimens to further research. A significant number of research studies, including 15 master of science theses, have been accomplished in UOP's Craniofacial Genetics Laboratory, with contributions by faculty, undergraduate and graduate students. It has been leading to a better understanding of etiology of nonsyndromic orofacial clefts. It has been learned that genetic factors and environmental factors are ethnicity specific and, in many places throughout the world, location-specific. Thus, a specific protocol for cleft prevention has to be worked out based on genetic and nutritional studies of each specific population group in order to be effective. This is our ultimate goal. PMID- 17087398 TI - Current clinical research in orthodontics: a perspective. AB - This essay explores briefly the approach of the Craniofacial Research Instrumentation Laboratory to the systematic and rigorous investigation of the usual outcome of orthodontic treatment in the practices of experienced clinicians. CRIL's goal is to produce a shareable electronic database of reliable, valid, and representative data on clinical practice as an aid in the production of an improved environment for truly evidence-based orthodontic treatment. PMID- 17087399 TI - Researching trust relations in health care: conceptual and methodological challenges--introduction. AB - PURPOSE: The aim of this paper is to provide a rationale for examining trust in health care. DESIGN/METHODOLOGY/APPROACH: Conducts a review of the literature of trust relations in health care that highlighted that most empirical research has addressed threats to patient-provider relationships and trust in health care systems from the patient's perspective, but studies in the organisational literature suggests that trust relations in the workforce, between providers and between providers and managers, may also influence patient-provider relationships and levels of trust. FINDINGS: Suggests that trust is not primarily dispositional or an individual attribute or psychological state, but is constructed from a set of inter-personal behaviors or from a shared identity. These behaviors are underpinned by sets of institutional rules, laws and customs. RESEARCH LIMITATIONS/IMPLICATIONS: This introductory paper has presented some evidence from an international, comparative study but there is the need for further, more detailed investigation into why trust relations may vary in different health care systems. ORIGINALITY/VALUE: This introductory paper provides a rationale for examining trust in health care and a context for the different elements of trust. PMID- 17087400 TI - Trust in health care: theoretical perspectives and research needs. AB - PURPOSE: This paper presents some key theoretical issues about trust, and seeks to demonstrate their relevance to understanding of, and research on, health systems. Although drawing particularly on empirical evidence from low- and middle income countries (LMICs), the paper aims to stimulate thinking across country settings. DESIGN/METHODOLOGY/APPROACH: Drawing both on conceptual literature and relevant empirical research from LMICs, the paper presents an argument about the role of trust within key health system relationships and identifies future research needs. FINDINGS: Theoretical perspectives on four questions are first discussed: what is trust and can it be constructed? Why does it matter to health systems? On what is it based? What are the dangers of trust? The relevance of these theoretical perspectives is then considered in relation to: understanding the nature of health systems; issues of equity and justice in health care; and policy and managerial priorities. The identified research needs are investigation of: the role of trusting workplace relationships as a source of non-financial incentives; the influence of trust over the operation of different forms of citizen-health system engagement; approaches to training trustworthy public managers; and the institutional developments required to sustain trustworthy behaviour within health systems. PRACTICAL IMPLICATIONS: The policy and management actions needed to strengthen health systems within LMICs, and elsewhere, include: recruitment of health workers that have the attitudes and capacity for moral understanding and motivation; training curriculae that develop such motivation; and developing the institutions (e.g., communication and decision-making practices, payment mechanisms) that can sustain trusting relationships across a health system. It is also important to recognise that distrust in some relationships may act to guard against the abuse of power. ORIGINALITY/VALUE: Although the notion of trust has become of increasing importance in health policy debates in high-income countries, it has received less attention in the context of LMICs. The papers adds to the very limited literature on trust in LMIC health systems and also opens new lines of thinking for those working in high income countries--particularly around the role of health systems in generating wider social value. PMID- 17087401 TI - Trust relations in health care: developing a theoretical framework for the "new" NHS. AB - PURPOSE: This paper seeks to address how and why trust relations in the NHS may be changing and presents a theoretical framework for exploring them in future empirical research. DESIGN/METHODOLOGY/APPROACH: This paper provides a conceptual analysis. It proposes that public and patient trust in health care in the U.K. appears to be shaped by a variety of factors. From a macro perspective, any changes in levels of public trust in health care institutions appear to derive partly from top-down policy initiatives that have altered the way in which health services are organised and partly from broader social and cultural processes. A variety of policy initiatives, including the introduction of clinical governance and the resulting use of performance management to scrutinise and change clinical activity, increasing patient choice and involvement in decision-making regarding their care, are examined for how they have changed the context for trust relations within the NHS. FINDINGS: It is argued that these policy initiatives have produced a new context for trust relations within the NHS, shifting the inter-dependence and distribution of power between patients, clinicians, and mangers and changing their vulnerability to each other and to health care institutions. The paper presents a theoretical framework based on current policy discourses which illustrates how new forms of trust relations may be emerging in this new context of health care delivery, reflecting a change in motivations for trust from affect based to cognition based trust as patients, clinicians and managers become more active partners in trust relations. The framework suggests that trust relations in all three types of relationship in the "new" modernised NHS might, in general, be particularly characterised by an emphasis on communication, providing information and the use of "evidence" to support decisions in a reciprocal, negotiated alliance. ORIGINALITY/VALUE: The paper examines the drivers for change in trust in health care relations in the U.K. and develops a theoretical framework for the emergence of new trust relations that can be subsequently explored through empirical research. PMID- 17087402 TI - Trust in the context of patient safety problems. AB - PURPOSE: This paper considers some implications of recent developments relating to patient safety for understandings of trust in health care contexts. DESIGN/METHODOLOGY/APPROACH: Conceptual analysis focusing on patients' trust in health care providers and health care providers' trust in patients. FINDINGS: Growing awareness of the scale of the problem of iatrogenic harm has prompted concerns that patients' trust in health care providers may be threatened and/or become inappropriate or dysfunctional. In principle, however, patients' trust may be both well placed and compatible with current understandings of safety problems and efforts to address these. Contemporary understandings of patient safety suggest that, to be deemed trustworthy, health care providers should make vigorous efforts to improve patient safety, be honest about safety issues, enable patients to contribute effectively to their own safety, and provide appropriate care and support after safety incidents. Patients who trust health care providers need not be ignorant of patient safety problems and may be vigilant in the course of their care. Iatrogenic harms do not necessarily reflect breeches of trust (not all such harms are yet preventable), and patients who are harmed might in some circumstances appropriately forgive and resume trusting. Health care providers may feel vulnerable to patients in several respects. From their perspective, trustworthy patients will act competently to optimise the outcomes of their health care efforts and to preserve health care providers' good reputations where those are justified. Providers' trust in patients may strengthen patients' trust in them and facilitate safety improvement work. ORIGINALITY/VALUE: Shows how, in principle, trust can be compatible with current understandings of patient safety issues and may enhance efforts to improve patient safety. PMID- 17087403 TI - Conceptualisations of trust in the organisational literature: some indicators from a complementary perspective. AB - PURPOSE: This paper evaluates the non-healthcare organisational literature on conceptualisations of trust. The aim of the paper is to review this diverse literature, and to reflect on the potential insights it might offer healthcare researchers, policy makers and managers. DESIGN/METHODOLOGY/APPROACH: A number of the key concepts that contribute to contrasting definitions of trust in the organisational literature are identified. FINDINGS: The paper highlights the heterogeneity of trust as an organisational concept. Aspects of trust that relate more specifically to non-healthcare settings are shown to have some potential relevance for healthcare. Five aspects of trust, considered to have particular significance to the changing face of the NHS, appear to offer scope for further exploration in healthcare settings. PRACTICAL IMPLICATIONS: The NHS continues to face changes to its organisational structures, both planned and unplanned. Healthcare providers will need to be alert to intra- and inter-organisational relationships, of which trust issues will form an inevitable part. Whilst it might be argued that the lessons offered by conceptualisations of trust within wider organisational settings have limitations, the paper demonstrates sufficient areas of overlap to encourage cross-fertilisation of ideas. ORIGINALITY/VALUE: The paper draws together previous research on a topic of increasing relevance to healthcare researchers, which has exercised management researchers for at least three decades. The paper acts as a guide to future research and practice. PMID- 17087404 TI - "You have to cover up the words of the doctor": the mediation of trust in interpreted consultations in primary care. AB - PURPOSE: This article explores issues of trust in narratives of interpreted consultations in primary health care. DESIGN/METHODOLOGY/APPROACH: The paper is based on empirical data from a qualitative study of accounts of interpreted consultations in U.K. primary care, undertaken in three north London boroughs. In a total of 69 individual interviews and two focus groups, narratives of interpreted consultations were sought from 18 service users, 17 professional interpreters, nine family member interpreters, 13 general practitioners, 15 nurses, eight receptionists, and three practice managers. The study collected and analysed these using a grounded theory approach and taking the story as the main unit of analysis. It applies a theoretical model that draws on three key concepts: Greener's taxonomy of trust based on the different "faces" of power in medical consultations; Weber's notion of bureaucratic vs traditional social roles; and Habermas' distinction between communicative and strategic action. FINDINGS: Trust was a prominent theme in almost all the narratives. The triadic nature of interpreted consultations creates six linked trust relationships (patient-interpreter, patient-clinician, interpreter-patient, interpreter clinician, clinician-patient and clinician-interpreter). Three different types of trust are evident in these different relationships--voluntary trust (based on either kinship-like bonds and continuity of the interpersonal relationship over time, or on confidence in the institution and professional role that the individual represents), coercive trust (where one person effectively has no choice but to trust the other, as when a health problem requires expert knowledge that the patient does not have and cannot get) and hegemonic trust (where a person's propensity to trust, and awareness of alternatives, is shaped and constrained by the system so that people trust without knowing there is an alternative). These different types of trust had important implications for the nature of communication in the consultation and on patients' subsequent action. RESEARCH LIMITATIONS/IMPLICATIONS: The methodological and analytic approach, potentially, has wider applications in the study of other trust relationships in health and social care. Practical implications - Quality in the interpreted consultation cannot be judged purely in terms of accuracy of translation. The critical importance of voluntary trust for open and effective communication, and the dependence of the latter on a positive interpersonal relationship and continuity of care, should be acknowledged in the design and funding of interpreting services and in the training of both clinicians, interpreters and administrative staff. ORIGINALITY/VALUE: This is the first study in which interpreted consultations have been analysed from a perspective of critical sociology with a particular focus on trust and power relations. PMID- 17087405 TI - Researching medical trust in the United States. AB - PURPOSE: This article reviews research in the U.S.A. bearing on trust in physicians and medical institutions. DESIGN/METHODOLOGY/APPROACH: This article provides a conceptual analysis, and general review of the literature. FINDINGS: Empirical research of medical trust is burgeoning in the U.S.A., and a fairly clear conceptual model of interpersonal physician trust has emerged. However, most studies focus on individual patients and their physicians, due to the highly individualistic attitudes that prevail in the U.S.A. Lacking are studies of more social dimensions of trust in broader medical institutions. A conceptual model of trust is presented to help draw these relevant distinctions, and to review the US literature. Also presented are the full set of trust scales, developed at Wake Forest University, which follow this conceptual model. These conceptual categories may differ, however, in other languages and cultures. ORIGINALITY/VALUE: The considerable body of research in the USA on patients' trust in individual physicians should help inform and focus international efforts to study social trust in medical institutions. PMID- 17087406 TI - Public trust in health care: a performance indicator? AB - PURPOSE: If public trust in health care is to be used as a performance indicator for health care systems, its measurement has to be sensitive to changes in the health care system. For this purpose, this study has monitored public trust in health care in The Netherlands over an eight-year period, from 1997 to 2004. The study expected to find a decrease in public trust, with a low point in 2002. DESIGN/METHODOLOGY/APPROACH: Since 1997, public trust in health care was measured through postal questionnaires to the "health care consumer panel". This panel consists of approximately 1500 households and forms a representative sample of the Dutch population. FINDINGS: Trust in health care and trust in hospitals did not show any significant trend. Trust in medical specialists displayed an upward trend. Trust in future health care, trust in five out of six dimensions of health care and trust in general practitioners actually did show a decrease. However, only for trust in macro level policies and trust in professional expertise this trend continued. For the remaining trust objects, after 1999 or 2000, an upward trend set in. RESEARCH IMPLICATIONS/LIMITATIONS: No support was found for our overall assumption. Explanations for the fact that trust did increase after 1999 or 2000 are difficult to find. On the basis of these findings the study questions whether the measure of public trust is sensitive enough to provide information on the performance of the health care system. ORIGINALITY/VALUE: The aim of this research is to study public trust in health care on its abilities to be used as a performance indicator for health care systems. PMID- 17087407 TI - Trust relations in health care: an agenda for future research. AB - PURPOSE: The aim of this paper is to draw together suggestions for future research from the papers and from the discussion that took place at the workshop. DESIGN/METHODOLOGY/APPROACH: The suggestions are summarised under four broad themes. FINDINGS: At an international workshop on trust organised by the U.K. MRC Health Services Research Collaboration there was broad agreement that trust was still a salient issue in diverse health care contexts. The workshop proceedings identified a number of important questions for empirical research and several key conceptual, theoretical and methodological questions relating to trust that need to be addressed in support of or alongside this. The collection of papers in this volume starts to address some of these questions. ORIGINALITY/VALUE: Considers trust relations in health care from patient, clinical, organisational and policy perspectives. PMID- 17087408 TI - How much longer can nurse be expected to cope and 'make do'? PMID- 17087409 TI - Keeping patients safe. Interview by Jeremy Benton. PMID- 17087410 TI - Nursing by numbers. AB - In the face of NHS budget cuts, nurses are being asked to justify their workforce numbers. Keith Hurst reviews some of the tools available for calculating staffing levels, examines their pros and cons, and discusses their application. PMID- 17087411 TI - Is it all in the mind? PMID- 17087412 TI - Enhancing the care of older people in the community. AB - This article describes a pilot project that aimed to enhance the care of older people who dialled 999 but did not require hospital admission. The project demonstrated how intermediate care teams and the ambulance service worked in partnership to avoid unnecessary hospital admission and to improve health and social care services for this patient group. PMID- 17087413 TI - Preparing to undertake research in the NHS: the novice's guide. AB - This article is aimed at nurses who are embarking on their first research project in the NHS, irrespective of whether the participants are members of staff, patients, carers or relatives. It describes the stages that the researcher needs to go through before beginning the research. PMID- 17087414 TI - Cognitive behavioural therapy for patients with anxiety and depression. AB - This article describes and explains the theoretical background of cognitive behavioural therapy. The evidence base for the cognitive behavioural treatment of a number of mental health problems is reported and the principles of effective treatment described. PMID- 17087415 TI - Epidural analgesia. PMID- 17087416 TI - Management of tissue excoriation in older patients with urinary or faecal incontinence. AB - This article discusses good skin care in relation to the management of incontinence. It outlines the structure and functions of the skin and describes how the skin changes as we age. It examines how incontinence can damage the skin and provides an overview of the current management methods that are used to prevent tissue excoriation. It also suggests an effective alternative that could be used if previous strategies have failed and the skin begins to breakdown, that is, the use of a silver regimen. PMID- 17087417 TI - The use of advanced biological and tissue-engineered wound products. AB - This article aims to reintroduce readers to biologically based and tissue engineered wound management products. Indications for their use in clinical practice are suggested. It also examines two well-established wound management interventions, topical negative pressure therapy and larval therapy, which could have a positive effect on the outcome of biologically based and tissue-engineered wound management products. PMID- 17087418 TI - Pulling power. PMID- 17087419 TI - To Russia with care. PMID- 17087420 TI - Choose the right company. PMID- 17087421 TI - Nesiritide for acute decompensated heart failure. PMID- 17087422 TI - Ultrasonography in the assessment of developmental dysplasia of the hip. PMID- 17087423 TI - The heart of women's health. PMID- 17087424 TI - Developmental dysplasia of the hip. AB - Developmental dysplasia of the hip refers to a continuum of abnormalities in the immature hip that can range from subtle dysplasia to dislocation. The identification of risk factors, including breech presentation and family history, should heighten a physician's suspicion of developmental dysplasia of the hip. Diagnosis is made by physical examination. Palpable hip instability, unequal leg lengths, and asymmetric thigh skinfolds may be present in newborns with a hip dislocation, whereas gait abnormalities and limited hip abduction are more common in older children. The role of ultrasonography is controversial, but it generally is used to confirm diagnosis and assess hip development once treatment is initiated. Bracing is first-line treatment in children younger than six months. Surgery is an option for children in whom nonoperative treatment has failed and in children diagnosed after six months of age. It is important to diagnose developmental dysplasia of the hip early to improve treatment results and to decrease the risk of complications. PMID- 17087425 TI - Hirschsprung's disease: diagnosis and management. AB - Hirschsprung's disease (congenital megacolon) is caused by the failed migration of colonic ganglion cells during gestation. Varying lengths of the distal colon are unable to relax, causing functional colonic obstruction. Hirschsprung's disease most commonly involves the rectosigmoid region of the colon but can affect the entire colon and, rarely, the small intestine. The disease usually presents in infancy, although some patients present with persistent, severe constipation later in life. Symptoms in infants include difficult bowel movements, poor feeding, poor weight gain, and progressive abdominal distention. Early diagnosis is important to prevent complications (e.g., enterocolitis, colonic rupture). A rectal suction biopsy can detect hypertrophic nerve trunks and the absence of ganglion cells in the colonic submucosa, confirming the diagnosis. Up to one third of patients develop Hirschsprung's-associated enterocolitis, a significant cause of mortality. Patients should be monitored closely for enterocolitis for years after surgical treatment of Hirschsprung's disease. With proper treatment, most patients will not have long-term adverse effects and can live normally. PMID- 17087426 TI - Information from your family doctor. Hirschsprung's disease: what you should know. PMID- 17087428 TI - Information from your family doctor. Cardiovascular disease: what you should know. PMID- 17087427 TI - Preventing cardiovascular disease in women. AB - Cardiovascular disease (CVD) has been the primary cause of death in women for almost a century, and more women than men have died of CVD every year since 1984. Although CVD incidence can be reduced by adherence to a heart-healthy lifestyle and detection and treatment of major risk factors, preventive recommendations have not been consistently or optimally applied to women. The American Heart Association guidelines for CVD prevention in women provide physicians with a clear plan for assessment and treatment of CVD risk and personalization of treatment recommendations. The emphasis of preventive efforts has shifted away from treatment of individual CVD risk factors in isolation toward assessment of a woman's overall or "global" CVD risk. In addition to accounting for the presence or absence of preexisting coronary heart disease or its equivalents (e.g., diabetes, chronic kidney disease), cardiovascular risk can be further calculated with the Framingham risk score, which is based on age, sex, smoking history, and lipid and blood pressure levels. Intervention intensity and treatment goals are tailored to overall risk, with those at highest risk receiving the most intense risk-lowering interventions. Women at high risk for CVD and without contraindications should receive aspirin, beta blockers, and an angiotensin converting enzyme inhibitor or angiotensin receptor blocker in addition to pharmacologic therapy for hyperlipidemia, hypertension, and diabetes. Women who already are at optimal or low risk for CVD should be encouraged to maintain or further improve their healthy lifestyle practices. Optimal application of these preventive practices significantly reduces the burden of death and disability caused by heart attack and stroke in women. PMID- 17087429 TI - Management of common opioid-induced adverse effects. AB - Opioid analgesics are useful agents for treating pain of various etiologies; however, adverse effects are potential limitations to their use. Strategies to minimize adverse effects of opioids include dose reduction, symptomatic management, opioid rotation, and changing the route of administration. Nausea occurs in approximately 25 percent of patients; prophylactic measures may not be required. Patients who do develop nausea will require antiemetic treatment with an antipsychotic, prokinetic agent, or serotonin antagonist. Understanding the mechanism for opioid-induced nausea will aid in the selection of appropriate agents. Constipation is considered an expected side effect with chronic opioid use. Physicians should minimize the development of constipation using prophylactic measures. Monotherapy with stool softeners often is not effective; a stool softener combined with a stimulant laxative is preferred. Sedation and cognitive changes occur with initiation of therapy or dose escalation. Underlying disease states or other centrally acting medications often will compound the opioid's adverse effects. Minimizing unnecessary medications and judicious use of stimulants and antipsychotics are used to manage the central nervous system side effects. Pruritus may develop, but it is generally not considered an allergic reaction. Antihistamines are the preferred management option should pharmacotherapy treatment be required. PMID- 17087430 TI - Management of head and neck injuries by the sideline physician. AB - Injuries to the head and neck are common in sports. Sideline physicians must be attentive and prepared with an organized approach to detect and manage these injuries. Because head and neck injuries often occur simultaneously, the sideline physician can combine the head and neck evaluations. When assessing a conscious athlete, the physician initially evaluates the neck for spinal cord injury and determines whether the athlete can be moved safely to the sideline for further evaluation. This decision is made using an on-field assessment of the athlete's peripheral sensation and strength, as well as neck tenderness and range of motion. If these evaluations are normal, axial loading and Spurling testing can be performed. Once the neck has been determined to be normal, the athlete can be assisted to the sideline for assessment of concussion symptoms and severity. This assessment should include evaluations of the athlete's reported symptoms, recently acquired memory, and postural stability. Injured athletes should be monitored with serial examinations, and those with severe, prolonged, or progressive findings require transport to an emergency department for further evaluation. PMID- 17087431 TI - Syncope: initial evaluation and prognosis. PMID- 17087432 TI - Screening for speech and language delay in preschool children. PMID- 17087433 TI - Bilateral knee pain. PMID- 17087435 TI - Information from your family doctor. Emotional health: what you should know. PMID- 17087434 TI - Information from your family doctor. Stress: how to cope with life's challenges. PMID- 17087436 TI - Information from your family doctor. Anxiety and panic: getting control over your feelings. PMID- 17087437 TI - Information from your family doctor. Depression: what you should know. PMID- 17087438 TI - Inclusion: the big delusion. PMID- 17087439 TI - Educational practice and research in education of deaf students science or art. PMID- 17087440 TI - Fingerspelling and sign language as alternative codes for reading and writing words for Chilean deaf signers. AB - The study examined the role of sign language and fingerspelling in the development of the reading and writing skills of deaf children and youth. Twenty six deaf participants (13 children, 13 adolescents), whose first language was Chilean Sign Language (CHSL), were examined. Their dactylic abilities were evaluated with tasks involving the reading and writing of dactylic and orthographic codes. The study included three experiments: (a) the identification of Chilean signs and fingerspelled words, (b) the matching of fingerspelled words with commercial logos, and (c) the decoding of fingerspelled words and the mapping of these words onto the writing system. The results provide convergent evidence that the use of fingerspelling and sign language is related to orthographic skills. It is concluded that fingerspelling can facilitate the internal representation of words and serve as a supporting mechanism for reading acquisition. PMID- 17087441 TI - Telemental health technology in deaf and general mental-health services: access and use. AB - Long-distance travel to provide mental health services for deaf people has implications for efficiency, safety, and equality of service. However, uptake of Telemental Health (TMH) has been slow in both deaf and general mental health services. A quantitative study was used to investigate access to TMH and whether staff confidence, experience, or demographics affect TMH use. It was concluded that staff in neither deaf mental health services nor general mental health services had adequate knowledge of or access to TMH. Staff expressed concerns over TMH's appropriateness in their work. Previous use of videoconferencing was assosciated significantly with confidence, but previous use of videophones was not. Neither staff in deaf services nor deaf staff were more experienced with or more confident about videoconferencing, whereas, within deaf services, deaf staff were significantly more confident about videophone use. Training implications are discussed. PMID- 17087442 TI - Prevalence of overweight among deaf children. AB - The study examined the prevalence of overweight cases in a sample of 151 deaf children aged 6-11 years. Participants were deaf students attending six elementary schools, both regular and special, in four states. Body mass index (BMI) was calculated using height and weight, plotted on the Center for Disease Control and Prevention (CDC) U.S. BMI-for-age growth charts, then compared to national values for same age and gender (CDC, 2006a, 2006b; National Center for Health Statistics, 2005). The results indicated that the prevalence of overweight deaf children aged 6-11 years was above the national percentage for same age and gender. A larger percentage of boys was overweight (24.7%) than girls (20.4%). After age 8 years, girls showed a consistent decrease in BMI with increasing age, a trend not demonstrated by boys. As a group, deaf children demonstrate a higher prevalence of overweight than national averages. PMID- 17087443 TI - Assessment of school functioning among Israeli Arab children with hearing loss in the primary grades. AB - The study examined school functioning of Israeli Arab children with hearing impairment (HI) who were included in regular education classrooms, in comparison to their classmates with normal hearing (NH). Ninety-three children (60 NH and 33 HI), grades 1-6, participated. Teachers evaluated the children using the Arabic version of the Screening Instrument for Targeting Educational Risks (SIFTER; Anderson, 1989); they also reported children's achievement levels in Arabic and mathematics. Results demonstrated that the HI children's functioning was lower than that of the NH children. Children with unilateral and minimal hearing losses had lower functioning than those with more severe hearing loss. As grade level increased, functioning decreased among HI children. Use of the SIFTER was beneficial in detecting children with difficulties. Regular, ongoing SIFTER use and appropriate follow-up on the results may better enable each child to perform optimally in class. PMID- 17087444 TI - Interaction between the teacher and the congenitally deafblind child. AB - Empirical data on the development of interaction, communication, and language in deafblind children is very rare. To fill this gap, a case study was conducted in which the interaction between a teacher and a deafblind boy age 3 years 4 months was analyzed. Sequential analysis of their interaction confirmed some general clinical impressions about interaction with deafblind children, and provided the basis for suggestions on how the interaction pattern might be changed. PMID- 17087445 TI - Improving mathematics teaching and learning experiences for hard of hearing students with wireless technology-enhanced classrooms. AB - Hard of hearing students usually face more difficulties at school than other students. A classroom environment with wireless technology was implemented to explore whether wireless technology could enhance mathematics learning and teaching activities for a hearing teacher and her 7 hard of hearing students in a Taiwan junior high school. Experiments showed that the highly interactive communication through the wireless network increased student participation in learning activities. Students demonstrated more responses to the teacher and fewer distraction behaviors. Fewer mistakes were made in in-class course work because Tablet PCs provided students scaffolds. Students stated that the environment with wireless technology was desirable and said that they hoped to continue using the environment to learn mathematics. PMID- 17087446 TI - An investigation of the need for sign language assessment in deaf education. AB - The attitudes of educators of the deaf and other professionals in deaf education concerning assessment of the use of American Sign Language (ASL) and other sign systems was investigated. A questionnaire was distributed to teachers in a residential school for the deaf in California. In addition to questions regarding the availability of sign language assessment at their schools, participants responded to items concerning their motivation to use a test for sign language measurement. Of the 100 distributed surveys, 85 were completed and returned. Results showed overwhelming agreement among respondents concerning the importance of sign language assessment, along with the need for tools that appropriately measure signing skills. PMID- 17087447 TI - In search of factors in deaf and hearing children's reading comprehension. AB - The present study examined whether specific item characteristics, such as mode of acquisition (MoA) of word meanings, make reading comprehension tests particularly difficult for deaf children. Reading comprehension data on nearly 13,000 hearing 7-to-12-year-olds and 253 deaf 7-to-20-year-olds were analyzed, divided across test levels from second to sixth grade (not necessarily corresponding to chronological age). Factor analyses across item scores suggested that, of the determinants studied, MoA--referring to the type of information (perceptual, linguistic, or both) used in word meaning acquisition--was the only factor that contributed significantly to deaf and hearing children's reading comprehension. For hearing children, MoA influenced item scores at the third- and fourth-grade levels. For the deaf children, MoA influenced item scores through the sixth-grade level. PMID- 17087448 TI - CJD: update for dental staff. AB - It is almost a decade since the recognition of the emergence of a new infectious disease termed variant Creutzfeldt-Jakob disease (vCJD) caused by prions (PrPTSE), abnormal variants of a normal human cell surface protein (PrP). This disease has a number of similarities to other forms of CJD--lethal disorders characterized by a prolonged incubation period, and progressive mental deterioration. In relation to oral tissues, PrPTSE have been found in neural, gingival, pulpal, lingual, lymphoreticular and salivary gland tissue in animal models. In both sporadic and variant CJD, PrPTSE is detectable in the trigeminal ganglion and, in vCJD, in lymphoreticular tissues, but infectivity has not been tested in other human oral tissues. CLINICAL RELEVANCE: PrPTSE is much more resistant to the common methods of inactivation than conventional pathogens, and it adheres avidly to steel whilst retaining its infectivity. Particular attention must be paid to cleaning and sterilizing re-usable dental instruments. Single-use devices, such as endodontic files and matrix bands, must never be re-used. Advice on the reprocessing of dental instruments used on known CJD patients must be obtained from local infection control teams. Research into effective methods of prion inactivation appears promising, although further work on the applicability to general dental practice is required. PMID- 17087449 TI - Treatment and maintenance of a dentate patient with 'radiation caries'. AB - Patients with xerostomia are presenting dental practitioners with challenges in caries control, long-term restoration and prosthodontic difficulties. In many cases, extraction may be the best option, but for younger, dentate patients, this may be inappropriate. This paper describes the management of a young partially dentate patient with severe xerostomia following irradiation of the salivary glands. Preventive and restorative management are discussed, together with treatment and healing of peri-radicular pathology. The case report demonstrates that long-term stabilization and management of caries and peri-radicular lesions are possible over a seven-year period for a patient with severe radiation caries. CLINICAL RELEVANCE: Many dental patients present with some degree of xerostomia due to age, side-effects of anti-hypertensive and psychotropic drugs and also as a side-effect of radiotherapy. General dental practitioners are ideally placed to monitor and provide early intervention for this highly caries-susceptible group of patients. With good patient motivation and professional support, tooth loss is not inevitable and this case report suggests strategies and demonstrates the clinical stages in the management of severe caries due to xerostomia. PMID- 17087450 TI - Common facial fractures: 3. Complications. AB - The final article of three, this paper highlights the complications that may arise either as a direct result of the fracture or the treatment of the fracture. CLINICAL RELEVANCE: Dentists may encounter and sometimes have to manage some of the complications of facial fractures or their treatment. PMID- 17087451 TI - IOTN (DHC): is it supported by evidence? AB - The strength of the evidence linking some common malocclusion traits (which are used as the basis for IOTN classification) with dental disease is reviewed. It is concluded that IOTN is not a reliable predictor of the potential impact of malocclusion on future dental health, and that alternative indices, which place greater emphasis on the aesthetic impairment and take account of patient perceptions, are more appropriate for assessing an individual's orthodontic treatment need. CLINICAL RELEVANCE: IOTN is likely to assume greater significance with the advent of the new GDS/PDS regulations pertaining to orthodontics. However, all practitioners, including those who commission specialist services, should be aware of its limitations, especially in relation to the Dental Health Component. PMID- 17087452 TI - The use of NSAIDs in dentistry: a case study of gastrointestinal complications. AB - NSAIDs are effective at reducing mild to moderate pain and are therefore commonly prescribed in dental practice. Dental practitioners should be familiar with their indications, contra-indications and side-effects.We present the case of a 28-year old male who had been taking two NSAIDs concurrently for dental pain which resulted in an acute upper gastrointestinal haemorrhage. CLINICAL RELEVANCE: General Dental Practitioners need to be aware of the possible life-threatening complications when prescribing NSAIDs. PMID- 17087453 TI - The management of abuse: 3. Domestic violence. AB - The role of the GDP and the dental team in the recognition and management of domestic violence is discussed. Information on the current legislation and protocols for referral are provided. CLINICAL RELEVANCE: This paper discusses domestic violence and offers information and practical advice for the dental team. PMID- 17087454 TI - Self-treatment using a power drill: a case report. AB - The intense pain of dental origin which patients suffer can be underestimated. This can lead them to self-harm or carry out self-treatment, sometimes with only the rudiments of how to achieve pain relief. A case is reported of a patient who attempted to relieve the intense pressure of an abscess arising in a maxillary cyst by using a home power drill. He drilled through the side of the tooth and drained the abscess but, unfortunately, fractured the drill. This was discovered at operation and meant the tooth had to be extracted. The patient had only received antibiotics prior to this for his abscess, with no drainage being undertaken. CLINICAL RELEVANCE: This case highlights the inappropriate use of antibiotics when drainage of an abscess is the correct treatment. It demonstrates the extreme measures some patients may take in order to relieve their symptoms by self-treating themselves physically, in addition to self-prescribed medication. PMID- 17087455 TI - Adhesive restoration of endodontically treated teeth--current research. AB - Restoring endodontically-treated teeth using adhesively cemented fibre posts is becoming popular in general practice. Clinical studies have been promising, however failures can occur, usually because of de-bonding of the post. Current research, aimed at identifying factors that could improve the adhesion of fibre posts, is described. CLINICAL RELEVANCE: Adhesively-retained fibre posts are becoming a popular choice for the restoration of endodontically treated teeth. Dental practitioners should be knowledgeable on the rationale behind these restorations, factors that can lead to failure, and ongoing developments. PMID- 17087456 TI - Physical signs for the general dental practitioner. PMID- 17087457 TI - Aspects of human disease. PMID- 17087458 TI - Freeze/thaw-induced embolism depends on nadir temperature: the heterogeneous hydration hypothesis. AB - Freeze/thaw-induced embolism was studied in leaves of field-grown snow gum (Eucalyptus pauciflora) subject to frequent morning frosts. Juvenile trees were grown in buried pots, brought to the laboratory at different stages of acclimation and subjected to simulated frost-freezes (at 2 degrees C h(-1)) to nadir temperatures of -3 or -6 degrees C, which snow gums commonly experience. Frost-frozen and subsequently thawed leaves were cryo-fixed to preserve the distribution of water and were then examined by cryo-scanning electron microscopy. No embolisms were found in leaves frozen to -3 degrees C and thawed. In contrast, 34% of vessels were embolized in thawed leaves that had been frozen to -6 degrees C. This difference was seen also in the extent of extracellular ice blocks in the mid-vein expansion zones in leaves frozen to -3 and -6 degrees C, which occupied 3 and 14% of the mid-vein area, respectively. While the proportion of embolism depended on nadir temperature, it was independent of season (and hence of acclimation state). From the observation that increased embolism at lower nadir temperature was related to the freeze-induced redistribution of water, we hypothesize that the dehydration of cell walls and cells caused by the redistribution exerts sufficient tension on xylem water to induce cavitation on thawing. PMID- 17087459 TI - Expression of a putative laccase gene, ZmLAC1, in maize primary roots under stress. AB - Laccases are multi-copper-containing glycoproteins and comprise a multi-gene family in plants. However, their physiological functions are still not well understood. We obtained sequence information for a putative laccase gene, ZmLAC1, from maize and studied ZmLAC1 expression in detail. The deduced ZmLAC1 protein was 70% identical to LpLAC5-4, a laccase from ryegrass. ZmLAC1 was expressed in leaves, stems and roots of maize seedlings. In unstressed maize primary roots, a higher ZmLAC1 transcript level was located in the basal region where cell elongation had ceased compared to the apical 5 mm of the roots where cells were rapidly dividing and elongating. A treatment with 300 mM NaCl resulted in a shortened root elongation zone (< 2 mm) and swelling in the apical 5 mm. Associated with the morphological change, the transcript level of ZmLACl was enhanced in the apical 5 mm, reaching a level similar to that in the basal region. Other abiotic stresses tested--such as 28.5% polyethylene glycol (PEG), which caused an inhibition of root elongation comparable to 300 mM NaCl--did not affect ZmLAC1 transcript level. Potential roles of ZmLAC1 in the roots responding to NaCl or other high concentration of salts are discussed. PMID- 17087460 TI - Supplementary ultraviolet-B irradiation reveals differences in stress responses between Arabidopsis thaliana ecotypes. AB - Irradiation of Arabidopsis thaliana ecotypes C24, Wassilewskija (Ws) and Columbia 0 (Col-0) with supplementary ultraviolet-A+B (UV-A+B) radiation revealed ecotype specific differences in expression of the gene for the pathogenesis-related protein PR-5. C24 showed an increased expression level of PR-5 (5- and 20-fold higher compared with Col-0 and Ws, respectively). Expression of other molecular markers such as CHS (encoding chalcone synthase), MEB5.2 [encoding a gene strongly up-regulated by ultraviolet-B (UV-B)] and PYROA [encoding a pyridoxine (Vitamin B6) biosynthesis enzyme] only showed slight differences between ecotypes. Oxidative stress during UVA+B exposure was monitored by staining for H2O2. This analysis also revealed important ecotype-specific differences. 'H2O2 hot spots' were found in C24, whereas an even distribution of H2O2 was found in Ws and Col-0. Necrotic lesions also appeared on C24 leaves after prolonged UV-B exposure. There was a reverse correlation between the H2O2 steady-state concentration and the PR-5 gene expression; Ws showed the highest level of H2O2 accumulation but the lowest expression level of the PR-5 gene. Furthermore, application of paraquat on the rosettes led to similar PR-5 expression and H2O2 accumulation patterns as were found after UV-A+B irradiation. The observed ecotypic differences were also reflected in a statistically significant UV-B dependent decrease in biomass, rosette size and leaf area for Ws, but not for C24 and Col-0. Our results show that a significant ecotype-specific genetic variability in general UV-B responses in Arabidopsis exists. Moreover, the signal transduction or gene regulation pathway for PR-5 differs from the other molecular markers used in this study. PMID- 17087461 TI - The characteristics of Ca -activated Cl- channels of the salt-tolerant charophyte Lamprothamnium. AB - The dependence of the Ca++-activated Cl- channels on potential difference (PD) was extracted from current-voltage (I/V) profiles recorded at the time of hypotonic regulation while the large conductance (G) K+ channels were blocked by tetraethylammonium (TEA). The total clamp current (I) was dominated by the Cl- I, i(Cl), with small contribution from the background I (i(background)). The i(Cl) was fitted by the Goldman-Hodgkin-Katz (GHK) model with enhanced PD dependence simulated by Boltzmann probability distributions. The i(background) was modelled by an empirical equation. The i(Cl) responded to PD changes within tens of milliseconds. The G maxima were located between -20 and -150 mV. The Cl- channel number and channel permeability parameter, N(Cl)P(Cl), decreased as a function of time in a hypotonic medium (from 0.45 x 10(-7) to 0.17 x 10(-7) ms(-1) in 19 min), with the positive half activation PD, V50+, shifting from +35 to -65 mV, and the negative half activation PD, V50-, shifting from -134 to -310 mV. The fitted Cl- concentration [Cl-]cyt at the time of hypotonic regulation indicated rapid equalization of vacuolar and cytoplasmic concentrations. Excellent data obtained under similar experimental conditions in a previous study enabled us to infer [Ca++]cyt influences on the Cl- channel characteristics. Thick sulphated polysaccharide mucilage, found on Lamprothamnium cells acclimated to more saline media, eliminated the activation of the i(Cl) at the time of the hypotonic regulation. This effect was reversed by the application of the enzyme heparinase. The characteristics of the i(Cl) were found to be consistent with a component of the excitation Is at the time of the action potential (AP). The short duration of the excitation transients was contrasted with that of the hypotonic regulation. The mechanisms for Cl- channel activation (and hence the Ca++ channel activation) were considered. PMID- 17087462 TI - Circumnutation of rice coleoptiles: its relationships with gravitropism and absence in lazy mutants. AB - Although circumnutation occurs widely in higher plants, its mechanism is little understood. The idea that circumnutation is based on gravitropism has long been investigated, but the reported results have been controversial. We used dark grown coleoptiles of rice (Oryza sativa L.) to re-investigate this issue. The following results supported the existence of a close relationship between gravitropism and circumnutation: (1) circumnutation disappears on a horizontal clinostat; (2) circumnutation is interrupted by a gravitropic response and re initiated at a definable phase after gravitropic curvature; (3) circumnutation can be re-established by submergence and a brief gravitropic stimulation in the coleoptiles that have stopped nutating in response to red light; and (4) lazy mutants show no circumnutation. In spite of these results, however, there were cases in which gravitropism and circumnutation could be separated. Firstly, the non-circumnutating lazy coleoptile showed nearly a wild-type level of gravitropic responsiveness in its upper half, although this part was an active site of both gravitropism and circumnutation in wild-type coleoptiles. Secondly, coleoptiles could nutate without overshooting the vertical when developing phototropic curvature. It is concluded that gravitropism influences, but it is not directly involved in the process of circumnutation. It is further suggested that a gravity signal, shared with gravitropism, contributes to the maintenance of circumnutation. PMID- 17087463 TI - Linking physiological processes with mangrove forest structure: phosphorus deficiency limits canopy development, hydraulic conductivity and photosynthetic carbon gain in dwarf Rhizophora mangle. AB - Spatial gradients in mangrove tree height in barrier islands of Belize are associated with nutrient deficiency and sustained flooding in the absence of a salinity gradient. While nutrient deficiency is likely to affect many parameters, here we show that addition of phosphorus (P) to dwarf mangroves stimulated increases in diameters of xylem vessels, area of conductive xylem tissue and leaf area index (LAI) of the canopy. These changes in structure were consistent with related changes in function, as addition of P also increased hydraulic conductivity (Ks), stomatal conductance and photosynthetic assimilation rates to the same levels measured in taller trees fringing the seaward margin of the mangrove. Increased xylem vessel size and corresponding enhancements in stem hydraulic conductivity in P fertilized dwarf trees came at the cost of enhanced mid-day loss of hydraulic conductivity and was associated with decreased assimilation rates in the afternoon. Analysis of trait plasticity identifies hydraulic properties of trees as more plastic than those of leaf structural and physiological characteristics, implying that hydraulic properties are key in controlling growth in mangroves. Alleviation of P deficiency, which released trees from hydraulic limitations, reduced the structural and functional distinctions between dwarf and taller fringing tree forms of Rhizophora mangle. PMID- 17087464 TI - Hydraulic design of pine needles: one-dimensional optimization for single-vein leaves. AB - Single-vein leaves have the simplest hydraulic design possible, yet even this linear water delivery system can be modulated to improve physiological performance. We determined the optimal distribution of transport capacity that minimizes pressure drop per given investment in xylem permeability along the needle for a given length without a change in total water delivery, or maximizes needle length for a given pressure difference between petiole and needle tip. This theory was tested by comparative analysis of the hydraulic design of three pine species that differ in the length of their needles [Pinus palustris (Engl.) Miller, approximately 50 cm; Pinus ponderosa Lawson & Lawson, approximately 20 cm and Pinus rigida Miller, approximately 5 cm]. In all three species, the distribution of hydraulic permeability was similar to that predicted by the optimum solution. The needles of P. palustris showed an almost perfect match between predicted and actual hydraulic optimum solution, providing evidence that vein design is a significant factor in the hydraulic design of pine leaves. PMID- 17087465 TI - Inorganic carbon acquisition in red tide dinoflagellates. AB - Carbon acquisition was investigated in three marine bloom-forming dinollagellates Prorocentrum minimum, Heterocapsa triquetra and Ceratium lineatum. In vivo activities of extracellular and intracellular carbonic anhydrase (CA), photosynthetic O2 evolution, CO2 and HCO3- uptake rates were measured by membrane inlet mass spectrometry (MIMS) in cells acclimated to low pH (8.0) and high pH (8.5 or 9.1). A second approach used short-term 14C-disequilibrium incubations to estimate the carbon source utilized by the cells. All three species showed negligible extracellular CA (eCA) activity in cells acclimated to low pH and only slightly higher activity when acclimated to high pH. Intracellular CA (iCA) activity was present in all three species, but it increased only in P. minimum with increasing pH. Half-saturation concentrations (K1/2) for photosynthetic O2 evolution were low compared to ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) kinetics. Moreover, apparent affinities for inorganic carbon (Ci) increased with increasing pH in the acclimation, indicating the operation of an efficient CO2 concentration mechanism (CCM) in these dinoflagellates. Rates of CO2 uptake were comparably low and could not support the observed rates of photosynthesis. Consequently, rates of HCO3- uptake were high in the investigated species, contributing more than 80% of the photosynthetic carbon fixation. The affinity for HCO3- and maximum uptake rates increased under higher pH. The strong preference for HCO3- was also confirmed by the 14C-disequilibrium technique. Modes of carbon acquisition were consistent with the 13C-fractionation pattern observed and indicated a strong species-specific difference in leakage. These results suggest that photosynthesis in marine dinoflagellates is not limited by Ci even at high pH, which may occur during red tides in coastal waters. PMID- 17087466 TI - The effect of drought on C and N stable isotopes in different fractions of leaves, stems and roots of sensitive and tolerant beech ecotypes. AB - Beech seedlings from 11 German climatic provenances were exposed to a realistically timed drought treatment in a greenhouse experiment. The stable isotope composition of carbon (C) and nitrogen (N) was analysed in pooled bulk material of roots, stems and leaves, as well as in the aqueous extracts and starch fractions. The delta 13C values increased in bulk samples (BS) of roots, stems and leaves by drought, although no leaf growth occurred during the experimental period. A clear drought effect on delta 13C in aqueous extracts was detected in leaves. In aqueous extracts of stems and roots as well as in starch fractions of all organs, abundance of delta 13C also tended to be increased by drought, but this effect was not statistically significant. For both delta 13C and delta 15N, enrichment was observed from the site of uptake/ source to the site of use/sink. A gradient for delta 13C in all fractions from leaves (-29.49, 28.89 and -27.85 per thousand) to stems (-28.81, -27.48 and -26.98 per thousand) and to roots (-27.60, -26.37 and -26.48 per thousand) was detected in BS, aqueous extracts and starch, respectively. An opposite gradient for delta 15N was found in BS: 1.59 per thousand, 1.84 per thousand and 3.05 per thousand in roots, stems and leaves, respectively. delta 15N was neither affected by drought in the BS nor in aqueous extracts, but an effect of provenance was observed. Particularly in roots and stems, drought-sensitive provenances showed the strongest shifts in delta 13C induced by drought and the lowest delta 15N values. In the present experiment, delta 13C values were more affected by the environmental factor drought, while delta 15N values were more affected by the genetic factor provenance. PMID- 17087467 TI - Long-term acclimatization of hydraulic properties, xylem conduit size, wall strength and cavitation resistance in Phaseolus vulgaris in response to different environmental effects. AB - Phaseolus vulgaris grown under various environmental conditions was used to assess long-term acclimatization of xylem structural characteristics and hydraulic properties. Conduit diameter tended to be reduced and 'wood' density (of 'woody' stems) increased under low moisture ('dry'), increased soil porosity ('porous soil') and low phosphorus ('low P') treatments. Dry and low P had the largest percentage of small vessels. Dry, low light ('shade') and porous soil treatments decreased P50 (50% loss in conductivity) by 0.15-0.25 MPa (greater cavitation resistance) compared with 'controls'. By contrast, low P increased P50 by 0.30 MPa (less cavitation resistance) compared with porous soil (the control for low P). Changes in cavitation resistance were independent of conduit diameter. By contrast, changes in cavitation resistance were correlated with wood density for the control, dry and porous soil treatments, but did not appear to be a function of wood density for the shade and low P treatments. In a separate experiment comparing control and porous soil plants, stem hydraulic conductivity (kh), specific conductivity (ks), leaf specific conductivity (LSC), total pot water loss, plant biomass and leaf area were all greater for control plants compared to porous soil plants. Porous soil plants, however, demonstrated higher midday stomatal conductance to water vapour (gs), apparently because they experienced proportionally less midday xylem cavitation. PMID- 17087468 TI - Interactions between the effects of atmospheric CO2 content and P nutrition on photosynthesis in white lupin (Lupinus albus L.). AB - Phosphorus (P) is a major factor limiting the response of carbon acquisition of plants and ecosystems to increasing atmospheric CO2 content. An important consideration, however, is the effect of P deficiency at the low atmospheric CO2 content common in recent geological history, because plants adapted to these conditions may also be limited in their ability to respond to further increases in CO2 content. To ascertain the effects of low P on various components of photosynthesis, white lupin (Lupinus albus L.) was grown hydroponically at 200, 400 and 750 micromol mol(-1) CO2, under sufficient and deficient P supply (250 and 0.69 microM P, respectively). Increasing growth CO2 content increased photosynthesis only under sufficient growth P. Ribulose 1,5-biphosphate carboxylase/oxygenase (Rubisco) content and activation state were not reduced to the same degree as the net CO2 assimilation rate (A), and the in vivo rate of electron transport was sufficient to support photosynthesis in all cases. The rate of triose phosphate use did not appear limiting either, because all the treatments continued to respond positively to a drop in oxygen levels. We conclude that, at ambient and elevated CO2 content, photosynthesis in low-P plants appears limited by the rate of ribulose biphosphate (RuBP) regeneration, probably through inhibition of the Calvin cycle. This failure of P-deficient plants to respond to rising CO2 content above 200 micromol mol(-1) indicates that P status already imposes a widespread restriction in plant responses to increases in CO2 content from the pre-industrial level to current values. PMID- 17087469 TI - Effects of chronic ozone exposure on gene expression in Arabidopsis thaliana ecotypes and in Thellungiella halophila. AB - Arabidopsis thaliana (At) ecotypes Columbia-0 (Col-0), Wassilewskija (WS), Cape Verde Islands (Cvi-0) and a relative, Thellungiella halophila (Th), were exposed to 20-25% over ambient ozone [O3] in a free air concentration enrichment (FACE) experiment (http://www.soyFACE. uiuc.edu), mirroring increases expected in the near future. Col-0 and WS accelerated development and developed lesions within 10 d under increased ozone, while Cvi-0 and Th grew slowly. RNAs were used in microarray hybridizations (Col-0-based 26 000 elements, 70-mer oligonucleotides). A two-step analysis of variance (ANOVA) model, including comparison with values obtained under [O3], was used for analyses. WS showed the greatest number of changes in gene expression in response to ozone. Th showed the least changes, suggesting that its expression state at [O3] was sufficient for resistance at increased ozone. Patterns observed in ambient air controls for Cvi-0 and Col-0 were most similar, while Th showed the greatest number of differences compared with the other controls. Compared with Col-0, however, Cvi-0 showed higher levels of expression of chaperones, receptor kinase-like and photosynthesis-related genes in ambient air. Cvi-0 exhibited ozone-mediated changes in a pathway involving AtSR, a homologue of the mammalian NF kappa B family of redox-sensitive transcription factors, changes in chaperones, WRKY and C2H2 proteins and antioxidants. WS displayed ozone-mediated decreases in the expression of two AtSR/NF kappa B family members, C2-domain proteins and genes associated with cell wall growth and changes in the expression of marker genes for programmed cell death (PCD), among them RCD1, a key regulator in this pathway. Microarray data were verified by reverse transcriptase (RT)-PCR. We relate O3-response diversity across the four lines to different responses among signaling and transcriptional response networks and differences in gene expression at [O3] levels. PMID- 17087470 TI - Winter acclimation of PsbS and related proteins in the evergreen Arctostaphylos uva-ursi as influenced by altitude and light environment. AB - The evergreen groundcover bearberry (Arctostaphylos uva-ursi [L.] Sprengel) was characterized over two successive years (2002-2004) from both sun-exposed and shaded sites at a montane ponderosa pine and subalpine forest community of 1900- and 2800-m-high altitudes, respectively. During summer, photosynthetic capacities and pre-dawn photosystem II (PSII) efficiency were similarly high in all four populations, and in winter, only the sun-exposed and shaded populations at 2800 m exhibited complete down-regulation of photosynthetic oxygen evolution capacity and consistent sustained down-regulation of PSII efficiency. This photosynthetic down-regulation at high altitude involved a substantial decrease in PSII components [pheophytin, D1 protein, oxygen evolving complex ([OEC)], a strong up regulation of several anti-early-light-inducible protein (Elip)- and anti-high light-inducible protein (Hlip)-reactive bands and a warm-sustained retention of zeaxanthin and antheraxanthin (Z + A). PsbS, the protein modulating the rapid engagement and disengagement of Z +A in energy dissipation, exhibited its most pronounced winter increases in the shade at 1900 m, and thus apparently assumes a greater role in providing rapidly reversible zeaxanthin-dependent photoprotection during winter when light becomes excessive in the shaded population, which remains photosynthetically active. It is attractive to hypothesize that PsbS relatives (Elips/Hlips) may be involved in sustained zeaxanthin-dependent photoprotection under the more extreme winter conditions at 2800 m. PMID- 17087471 TI - Photoprotection mutants of Arabidopsis thaliana acclimate to high light by increasing photosynthesis and specific antioxidants. AB - Biochemical and physiological acclimation to different light environments is crucial for plant growth and survival. In high light (HL), feedback de-excitation (qE) is a well-known photoprotective mechanism that dissipates excess excitation energy in the light-harvesting antenna of photosystem II (PSII) and relieves excitation pressure in the photosynthetic electron transport chain. The xanthophylls zeaxanthin (Z) and lutein (L) function in qE, but also have roles as antioxidants. Although several studies have shown that qE is important during short-term fluctuations in light intensity, here we show that it is not required for the growth of Arabidopsis thaliana in prolonged HL conditions in the laboratory. Mutants that are deficient in qE alone, qE and Z synthesis, or in qE, Z synthesis and also L synthesis were able to grow at 1800 micromol photons m(-2) s(-1) and exhibited no major symptoms of photooxidative stress. The mutants (and wild type) acclimated to HL by increasing photosynthetic capacity and decreasing light harvesting, which together rendered qE less important for photoprotection. At a metabolite level, the HL-grown mutants appeared to compensate for their remaining qE deficit with increased alpha-tocopherol and ascorbate levels compared to the wild type. The specificity of this response provides insight into the relationship between qE and the antioxidant network in plants. PMID- 17087472 TI - Mechanisms of microbially enhanced Fe acquisition in red clover (Trifolium pratense L.). AB - Soil microorganisms may play an important role in plant Fe uptake from soils with low Fe bioavailability, but there is little direct experimental evidence to date. We grew red clover, an Fe-efficient leguminous plant, in a calcareous soil to investigate the role of soil microbial activity in plant Fe uptake. Compared with plants grown in non-sterlie (NS) grown plants, growth and Fe content of the sterile(s) grown plants was significantly inhibited, but was improved by foliar application of Fe EDTA, indicating that soil microbial activity should play an important role in plant Fe acquisition. When soil solution was incubated with phenolic root exudates from Fe-deficient red clover, a few microbial species thrived while growth of the rest was inhibited, suggesting that the Fe-deficient (-Fe) root exudates selectively influenced the rhizosphere's microbial community. Eighty six per cent of the phenolic-tolerant microbes could produce siderophore [the Fe(III) chelator] under -Fe conditions, and 71% could secrete auxin-like compounds. Interestingly, the synthetic and microbial auxins (MAs) significantly enhanced the Ferric reduction system, suggesting that MAs, in addition to siderophores, are important to plant Fe uptake. Finally, plant growth and Fe uptake in sterilized soil were significantly increased by rhizobia inoculation. Root Fe-EDTA reductase activity in the -Fe plant was significantly enhanced by rhizobia infection, and the rhizobia could produce auxin but not siderophore under Fe-limiting conditions, suggesting that the contribution of nodulating rhizobia to plant Fe uptake can be at least partially attributed to stimulation of turbo reductase activity through nodule formation and auxin production in the rhizosphere. Based on these observations, we propose as a model that root exudates from -Fe plants selectively influence the rhizosphere microbial community, and the microbes in turn favour plant Fe acquisition by producing siderophores and auxins. PMID- 17087473 TI - Cysteines 449 and 459 modulate the reduction-oxidation conformational changes of ribulose 1.5-bisphosphate carboxylase/oxygenase and the translocation of the enzyme to membranes during stress. AB - The role of cysteines 449 (Cys449) and 459 (Cys459) from the large subunit (LS) of ribulose 1-5-bisphosphate carboxylase/oxygenase (Rubisco) in the reduction oxidation (redox) regulation of the enzyme was assessed by site-directed mutagenesis of these residues and chloroplast transformation of Chlamydomonas reinhardtii. In vitro studies indicated that mutations C449S, C459S or C449S/ C459S do not affect the activity and proteolytic susceptibility of the enzyme in the reduced state. However, when oxidized, the mutant enzymes differed from the wild type (WT), showing an increased resistance to inactivation and, in the case of the double mutant (DM), an altered structural conformation as reflected by the kinetics of proteolysis with subtilisin. The response of the DM strain to saline stress revealed that the absence of Cys449 and Cys459 intensifies Rubisco degradation and the covalent disulfide and non-disulfide polymerization of the enzyme in vivo. Saline stress also induced Rubisco translocation to a membrane (M) fraction that contained only covalently polymerized enzyme. Rubisco mobilization to this M fraction was enhanced also in the DM strain. Altogether, these results indicate that Cys449 and Cys459 participate in the modulation of the conformational changes promoted by oxidative modifications retarding processes related to the catabolism of the enzyme in vivo. PMID- 17087474 TI - Induction of systemic resistance in tomato by N-acyl-L-homoserine lactone producing rhizosphere bacteria. AB - N-acyl-L-homoserine lactone (AHL) signal molecules are utilized by Gram-negative bacteria to monitor their population density (quorum sensing) and to regulate gene expression in a density-dependent manner. We show that Serratia liquefaciens MG1 and Pseudomonas putida IsoF colonize tomato roots, produce AHL in the rhizosphere and increase systemic resistance of tomato plants against the fungal leaf pathogen, Alternaria alternata. The AHL-negative mutant S. liquefaciens MG44 was less effective in reducing symptoms and A. alternata growth as compared to the wild type. Salicylic acid (SA) levels were increased in leaves when AHL producing bacteria colonized the rhizosphere. No effects were observed when isogenic AHL-negative mutant derivatives were used in these experiments. Furthermore, macroarray and Northern blot analysis revealed that AHL molecules systemically induce SA- and ethylene-dependent defence genes (i.e. PR1a, 26 kDa acidic and 30 kDa basic chitinase). Together, these data support the view that AHL molecules play a role in the biocontrol activity of rhizobacteria through the induction of systemic resistance to pathogens. PMID- 17087475 TI - White lupin has developed a complex strategy to limit microbial degradation of secreted citrate required for phosphate acquisition. AB - White lupins (Lupinus albus L.) respond to phosphate deficiency by producing special root structures called cluster roots. These cluster roots secrete large amounts of carboxylates into the rhizosphere, mostly citrate and malate, which act as phosphate solubilizers and enable the plant to grow in soils with sparingly available phosphate. The success and efficiency of such a P-acquisition strategy strongly depends on the persistence and stability of the carboxylates in the soil, a parameter that is influenced to a large extent by biodegradation through rhizosphere bacteria and fungi. In this study, we show that white lupin roots use several mechanisms to reduce microbial growth. The abundance of bacteria associated with cluster roots was decreased at the mature state of the cluster roots, where a burst of organic acid excretion and a drastic pH decrease is observed. Excretion of phenolic compounds, mainly isoflavonoids, induced fungal sporulation, indicating that vegetative growth, and thus potential citrate consumption, is reduced. In addition, the activity of two antifungal cell wall degrading enzymes, chitinase and glucanase, were highest at the stage preceding the citrate excretion. Therefore, our results suggest that white lupin has developed a complex strategy to reduce microbial degradation of the phosphate solubilizing agents. PMID- 17087476 TI - Non-steady state effects in diurnal 180 discrimination by Picea sitchensis branches in the field. AB - We report diurnal variations in 18O discrimination (18 delta) during photosynthesis (18 delta A) and respiration (18 delta R) of Picea sitchensis branches measured in branch chambers in the field. These observations were compared with predicted 18 delta (18 delta pred) based on concurrent measurements of branch gas exchange to evaluate steady state and non-steady state (NSS) models of foliage water 18O enrichment for predicting the impact of this ecosystem on the Delta 18O of atmospheric CO2. The non-steady state approach substantially improved the agreement between 18 delta pred and observed 18 delta (18 delta obs) compared with the assumption of isotopic steady state (ISS) for the Delta 18O signature of foliage water. In addition, we found direct observational evidence for NSS effects: extremely high apparent 18 delta values at dusk, dawn and during nocturnal respiration. Our experiments also show the importance of bidirectional foliage gas exchange at night (isotopic equilibration in addition to the net flux). Taken together, neglecting these effects leads to an underestimation of daily net canopy isofluxes from this forest by up to 30%. We expect NSS effects to be most pronounced in species with high specific leaf water content such as conifers and when stomata are open at night or when there is high relative humidity, and we suggest modifications to ecosystem and global models of delta 18O of CO2. PMID- 17087477 TI - Heterogeneity of plant mitochondrial responses underpinning respiratory acclimation to the cold in Arabidopsis thaliana leaves. AB - In this study, we investigated whether changes in mitochondrial abundance, ultrastructure and activity are involved in the respiratory cold acclimation response in leaves of the cold-hardy plant Arabidopsis thaliana. Confocal microscopy [using plants with green fluorescence protein (GFP) targeted to the mitochondria] and transmission electron microscopy (TEM) were used to visualize changes in mitochondrial morphology, abundance and ultrastructure. Measurements of respiratory flux in isolated mitochondria and intact leaf tissue were also made. Warm-grown (WG, 25/ 20 degrees C day/night), 3-week cold-treated (CT) and cold-developed (CD) leaves were sampled. Although CT leaves exhibited some evidence of acclimation (as evidenced by higher rates of respiration at moderate measurement temperatures), it was only the CD leaves that were able to re establish respiratory flux within the cold. Associated with the recovery of respiratory flux in the CD leaves were: (1) an increase in the total volume of mitochondria per unit volume of tissue in epidermal cells; (2) an increase in the ratio of cristae to matrix within mesophyll cell mitochondria; and (3) an increase in the capacity of the energy-producing cytochrome pathway in mitochondria isolated from whole leaf homogenates. Regardless of growth temperature, we found that contrasting cell types exhibited distinct differences in mitochondrial ultrastructure, morphology and abundance. Collectively, our data demonstrated the diversity and tissue-specific nature of mitochondrial responses that underpin respiratory acclimation to the cold, and revealed the heterogeneity of mitochondrial structure and abundance that exists within leaves. PMID- 17087478 TI - Comparative transcriptome analysis of toxic metal responses in Arabidopsis thaliana and the Cd(2+)-hypertolerant facultative metallophyte Arabidopsis halleri. AB - Toxic effects of both essential and non-essential heavy metals are well documented in plants. Very little is known, however, about their modes of toxicity, about tolerance mechanisms and the signalling cascades involved in mediating transcriptional responses to toxic metal excess. We analysed transcriptome changes upon Cd2+ and Cu2+ exposure in roots of Arabidopsis thaliana and the Cd(2+)-hypertolerant metallophyte Arabidopsis halleri. Particularly, three categories of genes were identified with the help of this comparative approach: (1) common responses, which might indicate stable and functionally relevant changes conserved across plant species; (2) metallophyte specific responses as well as transcripts differentially regulated between the two species, representing candidate genes for Cd2+ hypertolerance; and (3) those specifically responsive to Cd2+ and therefore indicative of toxicity mechanisms or potentially involved in signalling cascades. Our data define, for instance, Arabidopsis core responses to Cd2+ and Cu2+. In addition, they suggest that Cd2+ exposure very rapidly results in apparent Zn deficiency, and they show the existence of highly specific Cd2+ responses and distinct signalling cascades. Array results were independently confirmed by real-time quantitative PCR, thereby further validating cross-species transcriptome analysis with oligonucleotide microarrays. PMID- 17087479 TI - A re-examination of the minor role of unstirred layers during the measurement of transport coefficients of Chara corallina internodes with the cell pressure probe. AB - The impact of unstirred layers (USLs) during cell pressure probe experiments with Chara corallina internodes has been quantified. The results show that the hydraulic conductivity (Lp) measured in hydrostatic relaxations was not significantly affected by USLs even in the presence of high water flow intensities ('sweep-away effect'). During pressure clamp, there was a reversible reduction in Lp by 20%, which was explained by the constriction of water to aquaporins (AQPs) in the C. corallina membrane and a rapid diffusional equilibration of solutes in arrays where water protruded across AQPs. In osmotic experiments, Lp, and permeability (Ps) and reflection (sigma s) coefficients increased as external flow rate of medium increased, indicating some effects of external USLs. However, the effect was levelling off at 'usual' flow rates of 0.20-0.30 m s(-1) and in the presence of vigorous stirring by air bubbles, suggesting a maximum thickness of external USLs of around 30 microm including the cell wall. Because the diameters of internodes were around 1 mm, internal USLs could have played a significant or even a dominating role, at least in the presence of the rapidly permeating solutes used [acetone, 2-propanol and dimethylformamide (DMF)]. A comparison of calculated (diffusion kinetics) and of measured permeabilities indicated an upper limit of the contribution of USLs for the rapidly moving solute acetone of 29%, and of 15% for the less rapidly permeating DME The results throw some doubt on recent claims that in C. corallina, USLs rather than the cell membrane dominate solute uptake, at least for the most rapidly moving solute acetone. PMID- 17087480 TI - Dynamics of stomatal water relations following leaf excision. AB - We examined the stomatal response to leaf excision in an evergreen woody shrub, Photinia x fraseri, using a novel combination of gas exchange, traditional water relations and modelling. Plants were kept outdoors in mild winter conditions (average daily temperature range: -1 to 12 degrees C) before being transferred to a glasshouse (temperature range: 20-30 degrees C) and allowed to acclimate for different periods before experiments. 'Glasshouse plants' were acclimated for at least 9 d, and 'outdoor plants' were acclimated for fewer than 3 d before laboratory gas exchange experiments. The transient stomatal opening response to leaf excision was roughly twice as long in outdoor plants as in glasshouse plants. To elucidate the reason for this difference, we inferred variables of stomatal water relations (epidermal and guard cell turgor pressures and guard cell osmotic pressure: Pe, Pg and pi g, respectively) from stomatal conductance (gs) and bulk leaf water potential (psi l), using a hydromechanical model of gs. psi l was calculated from cumulative post-excision transpirational water loss using empirical relationships between psi l and relative water content obtained on similar leaves. Inferred Pg and Pe both declined immediately after leaf excision. Inferred pi g also declined after a lag period. The kinetics of pi g adjustment after the lag were similar in outdoors and glasshouse plants, but the lag period was much longer in outdoor plants. This suggests that the longer transient opening response in outdoor plants resulted from slower induction, not slower execution, of guard cell osmoregulation. We discuss the implications of our results for the mechanism of short-term stomatal responses to hydraulic perturbations, for dynamic modelling of gs and for leaf water status regulation. PMID- 17087481 TI - Direct in situ measurement of cell turgor in grape (Vitis vinifera L.) berries during development and in response to plant water deficits. AB - Vitis vinifera L. berries are non-climacteric fruits that exhibit a double sigmoid growth pattern, and at the point known as 'veraison', which is just before the beginning of the second period of rapid fruit growth, these berries undergo several abrupt physiological changes. Cell pressure probe was used to examine the in situ turgor (P) of cells in the mesocarp during berry development and in response to plant water deficits. Initial tests comparing attached and detached berries demonstrated that cell P was stable for up to 48 h after detachment from the vine, provided that water loss from the berry was prevented. Cell P at pre-dawn was on the order of 0.25 MPa pre-veraison (PreV) and was reduced by an order of magnitude to 0.02 MPa post veraison (PostV). Cell P declined slightly but significantly with depth from the berry surface PreV, but not PostV. When water was withheld from potted vines, cell P declined about 0.2 Mpa, as pre-dawn vine water potential declined about 0.6 MPa over 12 d, whereas cell P was completely insensitive to a 1.10 MPa decrease in pre-dawn vine water potential after veraison. Rewatering of stressed plants also resulted in a 24 h recovery of cell P before, but not after veraison. The substantial decline in cell P around veraison is consistent with the decline in berry firmness that is known to occur at this time, and the PostV insensitivity of P to changes in vine water status is consistent with current hypotheses that the PostV berry is hydraulically isolated from the vine. The fact that a measurable P of about 0.02 MPa and typical cell hydraulic/osmotic behaviour were exhibited in PostV berries, however, indicates that cell membranes remain intact after veraison, contrary to many current hypotheses that veraison is associated with a general loss of membrane function and cellular compartmentation in the grape berry. We hypothesize that cell P is low in the PostV berry, and possibly other fleshy fruits, because of the presence of regulated quantities of apoplastic solutes. PMID- 17087482 TI - Modification of intracellular levels of glutathione-dependent formaldehyde dehydrogenase alters glutathione homeostasis and root development. AB - Glutathione (GSH)-dependent formaldehyde dehydrogenase (FALDH) is a highly conserved medium-chain dehydrogenase reductase and the main enzyme that metabolizes intracellular formaldehyde in eukaryotes. It has been recently shown that it exhibits a strong S-nitrosoglutathione (GSNO) reductase activity and could be a candidate to regulate NO-signalling functions. However, there is a lack of knowledge about the tissue distribution of this enzyme in plants. Here, we have studied the localization and developmental expression of the enzyme using immunolocalization and histochemical activity assay methods. We conclude that FALDH is differentially expressed in the organs of Arabidopsis thaliana mature plants, with higher levels in roots and leaves from the first stages of development. Spatial distribution of FALDH in these two organs includes the main cell types [epidermis (Ep) and cortex (Cx) in roots, and mesophyll in leaves] and the vascular system. Arabidopsis thaliana mutants with modified levels of FALDH (both by over- and under-expression of the FALDH-encoding gene) show a significant reduction of root length, and this phenotype correlates with an overall decrease of intracellular GSH levels and alteration of spatial distribution of GSH in the root meristem. Tansgenic roots are partially insensitive to exogenous GSH, suggesting an inability to detect reduction oxidation (redox) changes of the GSH pool and/or maintain GSH homeostasis. PMID- 17087483 TI - In vivo localization of manganese in the hyperaccumulator Gossia bidwillii (Benth.) N. Snow & Guymer (Myrtaceae) by cryo-SEM/EDAX. AB - Gossia bidwillii (Myrtaceae) is a manganese (Mn)-hyperaccumulating tree native to subtropical eastern Australia. It typically contains foliar Mn levels in excess of 1% dry weight. However, in G. bidwillii and other Mn-hyperaccumulating species, the cellular and subcellular localization of Mn has not been measured. Quantitative in vivo cryo-scanning electron microscopy (SEM)/energy dispersive X ray analysis (EDAX) was used to localize Mn and other elements in tissue collected from mature trees growing in a natural population. Cryo-SEM showed that the leaf mesophyll is differentiated as a double-layer palisade mesophyll above spongy mesophyll. Transmission electron microscopy (TEM) revealed that the palisade and epidermal cells are highly vacuolated. EDAX data were used to estimate in situ vacuolar Mn concentrations of all cell types in fresh cryo-fixed leaf tissues. The highest average vacuolar Mn concentration of over 500 mM was found in the upper-layer palisade mesophyll, while the lowest concentration of around 100 mM was found in the spongy mesophyll. Qualitative in vivo cryo SEM/EDAX was employed to further investigate the spatial distribution of Mn in fresh leaf tissues and young bark tissue, which was also found to have a high Mn concentration. It is concluded that Mn distribution in G. bidwillii is quantitatively different to metal distribution in other hyperaccumulating species where the highest localized concentrations of these elements occur in non photosynthmetic tissues such as epidermal cells and associated dermal structures including trichomes and leaf hairs. PMID- 17087484 TI - Power and parenting assessments: the intersecting levels of culture, race, class and gender. AB - This article uses a case example to highlight some of the complexities of working with minority ethnic families. It addresses the dilemmas that exist for therapists and other professionals when carrying out cross-cultural assessments of parenting. In this article, we will examine how professionals' constructions impacted on the assessment process with regard to family life, culture and mental health. We will draw out implications for clinical practice in this area. PMID- 17087485 TI - Exploring 'clinical judgement': how do child and adolescent mental health professionals decide whether a young person needs individual psychotherapy? AB - This article reports on a small-scale, qualitative study based in a single CAMHS team, exploring the decision-making process used by mental health professionals when deciding whether or not to refer a child for one particular modality of treatment--individual psychotherapy. Building on an initial audit of child psychotherapy referrals within the team, this study analyses a small number of in depth interviews with members of the multidisciplinary CAMHS team. Three themes emerged from the analysis, suggesting that referral-making decisions depend on wider conceptions of the particular modality of therapy, particular features of the child and family referred (but not reducible to diagnostic categories), and the stage at which therapeutic work with the family has reached at the time of referral. The implications of these findings are discussed specifically in relation to child psychotherapy, and more generally in relation to evidence-based practice. The authors conclude by arguing for the use of more clinically relevant research methodologies within evidence-based research. PMID- 17087486 TI - Are peer support groups for adolescents with Asperger's syndrome helpful? AB - There is very little documentation dealing with peer support groups for people with Asperger's syndrome, and especially about groups for adolescents. This article gives a description of three such groups that were run in parallel with support groups for parents. The great majority of the participants completed the course of the group. In a consumer satisfaction survey, the majority of the participants (76.5%) and nearly all of their parents (95%) rated satisfaction with the group meetings as good or very good. These responses indicate that adolescents with Asperger's syndrome and their parents perceive peer support groups as helpful. PMID- 17087487 TI - Development of multi-agency referral pathways for attention-deficit hyperactivity disorder, developmental coordination disorder and autistic spectrum disorders: reflections on the process and suggestions for new ways of working. AB - This article summarizes the rationale for multi-agency working when assessing children and young people with suspected neurodevelopmental disorders such as attention-deficit hyperactivity disorder, developmental coordination disorder and autistic spectrum disorders. Details of referral pathways that have been drawn up for each separate disorder are given and the process through which multi-agency agreement was reached is described. Key features thought to be common to all three pathways are discussed and factors thought to have contributed to successful multi-agency working highlighted. The work is described in the context of existing national policy documents in the UK. PMID- 17087488 TI - Associations between maternal childrearing cognitions and conduct problems in young children. AB - This study investigated maternal childrearing cognitions associated with ineffective parenting practices, using the Parental Childrearing Cognitions Questionnaire (PCCQ). Intergenerational transmission of parenting problems and cognitions was investigated using the Parental Bonding Instrument (PBI), which measures over-protectiveness and care experienced by parents in childhood. As a new measure, the PCCQ's psychometric properties were evaluated and found to be robust in terms of internal consistency and test-retest reliability. Seventy-four mothers of 3-5-year-old boys (23 clinical: boys referred with conduct problems; 51 control) completed the PCCQ, Revised Rutter Parent Scale for Preschool Children and PBI. PCCQ scores were significantly higher for clinical group mothers, and significantly correlated with both Parent and Teacher Rutter Child problem scores. There were significant correlations between PBI overprotection (positive) and PBI care (negative) scores and Rutter Parent and PCCQ scores. Clinical group mothers rated lower care and higher overprotection, from both parents, in their own childhoods. Results are consistent with a hypothesized model of intergenerational transmission of parenting problems, whereby experiences of low care and high overprotection in childhood predispose mothers to a dysfunctional 'set' of parenting cognitions, impairing maternal capacity to provide sensitive responses to challenging child behaviours. PMID- 17087489 TI - 'Watching from the stairs': towards an evidence-based practice in work with child witnesses of domestic violence. AB - This article describes the current practice of work with children who have witnessed domestic violence in the UK and North America. It examines this practice in the light of evidence of effectiveness and in the light of research that explains how witnessing domestic violence affects children. Finally it proposes guidelines for an evidenced-based practice which is built on this research. PMID- 17087490 TI - A follow-up study of chronic fatigue syndrome in children and adolescents: symptom persistence and school absenteeism. AB - This is a follow-up study of 28 young people aged between 7 and 17 meeting the Oxford criteria for the diagnosis of chronic fatigue syndrome treated in a specialist paediatric/psychiatric service. Retrospective case note analysis revealed a wide range and duration of symptoms together with high levels of school absenteeism prior to the diagnosis. The mean follow-up interval after discharge from the specialist service was 3 years and although most of the young people regarded themselves as fully recovered by this time, improvement was variable and about one third were still experiencing disabling symptoms. The illness had impacted on the education or career plans of all the young people to some extent with 15 experiencing difficulty returning to school. This article highlights the need for early recognition and diagnosis of chronic fatigue syndrome in young people and the importance of continuing paediatric support to reduce symptom persistence in the sensitive recovery period. Maintaining school attendance by close liaison between health and education services both before and after diagnosis and treatment is also vital if long-term morbidity is to be reduced. PMID- 17087491 TI - Children's key concerns: piloting a qualitative approach to understanding their experience of mental health care. AB - This article reports the piloting of an approach to apply the principles of child involvement to service experience research. The approach aimed to systematically explore the service concerns of children who had received mental health care. Eleven children who had previously attended mental health services took part in focus groups to discuss their experience. Thematic content analysis of transcribed interviews revealed 13 themes that potentially could help to identify children's key concerns and clarify the notion of child-centred care. The themes suggested the potential importance of children's expectations about therapy, and their concerns relating to the process, content and outcome may be important to their experience of care. The possible implications for child-centred clinical practice and service provision are discussed. PMID- 17087492 TI - The development of a new day treatment program for older children (8-11 years) with behavioural problems: the GoZone. AB - The main aim of this article is to describe the development of a new day treatment program for older children (8-11 years) with behavioural problems. The article outlines the content of the program and it also sets out the rationale behind the development of the new day service. The day program involves therapeutic and educational input and children attend the program two days a week for one academic term (10-13 weeks). Therapeutic input focuses on improving functioning in relation to a number of developmental processes that are known to be linked to the development of problem behaviour. These include improving emotional competence, dealing with peer relationship problems and interpersonal difficulties, and changing negative patterns of thinking about the self and others. The GoZone team also attempt to work collaboratively with the children's families and schools. A preliminary investigation of the effectiveness of the program is also reported. Parents and teachers completed the Strengths and Difficulties Questionnaire (SDQ) pre- and posttreatment. Findings showed that over the course of treatment parents reported a significant decrease in overall levels of emotional and behavioural problems and also reported a significant decrease in levels of emotional symptoms and peer problems. However, no significant changes in emotional and behavioural functioning were reported by teachers at school over the course of treatment. Potential ways of boosting the magnitude of positive change achieved by the new day treatment program are discussed. PMID- 17087493 TI - The demise of long-term work: is the clock ticking too quickly? PMID- 17087494 TI - Thermodynamic basis for redox regulation of the Yap1 signal transduction pathway. AB - The Yap1 oxidative stress signal transduction pathway found in Saccharomyces cerevisiae is redox-regulated. We have examined the thermodynamic basis of the disulfide/dithiol couples that are involved in the regulation of this pathway. The oxidized form of the Yap1 redox domain (Yap1-RD) fragment, derived from the Yap1 transcription factor, contains two disulfide bonds, one between Cys303 and Cys598 and one between Cys310 and Cys629. Oxidation-reduction titrations reveal the presence of two separate two-electron redox couples in Yap1-RD, with redox midpoint potentials (E(m)) of -155 and -330 mV, respectively, at pH 7.0. We measured E(m) values of -275 and -265 mV for the two cytoplasmic S. cerevisiae thioredoxins, Trx1 and Trx2, respectively, both at pH 7.0. Last, we measured an E(m) value of -255 mV for the Cys36-Cys82 disulfide bond at pH 6.0 in the glutathione peroxidase-like enzyme, oxidant receptor protein (Orp1). We were unable to obtain satisfactory redox titration data for Orp1 at pH 7.0, but if the redox-active disulfide of Orp1 exhibits the -59 mV per pH unit dependence for E(m) typical of protein disulfides in this pH region, an E(m) value of -315 mV can be estimated for Orp1 at pH 7.0 by extrapolation. Together, these data suggest that, at physiological ratios of Trx(ox)/Trx(red), the reduction of both the E(m) = -315 mV disulfide of Orp1 and the E(m) = -330 mV disulfide of Yap1 by either Trx1 or Trx2 would be thermodynamically possible. PMID- 17087495 TI - Secretory proteins as potential semiochemical carriers in the horse. AB - Two soluble proteins were isolated as major secretory products of horse sweat and of the parotid gland and characterized for structural and functional properties. The first protein, lipocalin allergen EquC1, was characterized for its glycosylation sites and bound glycosidic moieties. Only one (Asn53) of the two putative glycosylation sites within the sequence was post-translationally modified; a different glycosylation pattern was determined with respect to data previously reported. When purified from horse sweat, this protein contained oleamide and other organic molecules as natural ligands. Ligand binding experiments indicated good protein selectivity toward volatile compounds having a straight chain structure of 9-11 carbon atoms, suggesting a role of this lipocalin in chemical communication. The second protein, here reported for the first time in the horse, belongs to the group of parotid secretory proteins, part of a large superfamily of binding proteins whose function in most cases is still unclear. This protein was sequenced and characterized for its post-translational modifications. Of the three cysteine residues present, two were involved in a disulfide bridge (Cys155-Cys198). A model, built up on the basis of similar proteins, indicated a general fold characterized by the presence of a long hydrophobic barrel. Binding experiments performed with a number of different organic compounds failed to identify ligands for this protein with a well-defined physiological role. PMID- 17087496 TI - The Drosophila class B scavenger receptor NinaD-I is a cell surface receptor mediating carotenoid transport for visual chromophore synthesis. AB - The blind Drosophila mutant ninaD lacks the visual chromophore. Genetic evidence that the molecular basis is a defect in carotenoid uptake which causes vitamin A deficiency exists. The ninaD gene encodes a scavenger receptor that is significantly homologous in sequence with the mammalian scavenger receptors SR-BI (scavenger receptor class B type I) and CD36 (cluster determinant 36), yet NinaD has not been characterized in functional detail. Therefore, we established a Drosophila S2 cell culture system for biochemically characterizing the ninaD gene products. We show that the two splice variant isoforms encoded by ninaD exhibit different subcellular localizations. NinaD-I, the long protein variant, is localized at the plasma membrane, whereas the short variant, NinaD-II, is localized at intracellular membranes. Only NinaD-I could mediate the cellular uptake of carotenoids from micelles in this cell culture system. Carotenoid uptake was concentration-dependent and saturable. By in vivo analyses of different mutant and transgenic fly strains, we provide evidence of an essential role of NinaD-I in the absorption of dietary carotenoids to support visual chromophore synthesis. Moreover, our analyses suggest a role of NinaD-I in tocopherol metabolism. Even though Drosophila is a sterol auxotroph, we found no evidence of a contribution of NinaD-I to the uptake of these compounds. Together, our study establishes an evolutionarily conserved connection between class B scavenger receptors and the numerous functions of fat soluble vitamins in animal physiology. PMID- 17087497 TI - CRP subunit association and hinge conformation changes in response to cAMP binding: analysis of C-helix cysteine-substituted CRP. AB - We investigated the characteristics of 13 CRP variants having cysteine substituted at positions 113, 115, 116, 117, 118, 120, 122, 124, 126, 127, 129, 130, or 131, positions that span the length of the CRP C alpha-helix. Under reducing conditions, the WT and all Cys-substituted forms of CRP migrated as 23.5 kDa CRP monomer species on SDS-PAGE gels. In the absence of a reductant, 9 of 13 Cys-substituted forms of CRP including the L113C, S117C, M120C, L124C, V126C, T127C, E129C, K130C, and V131C CRP contained protein that migrated as 47 kDa CRP dimer species on SDS-PAGE gels. CNBr digestion of the protein preparations followed by MALDI-TOF MS analysis of the peptide fragments showed these 47 kDa species to be CRP dimers that originated from disulfide bonds formed between positional-pair C alpha-helix Cys residues. The ratio of monomer CRP and disulfide cross-linked CRP within a Cys-substituted CRP preparation was found to be independent of cAMP for Cys-substituted CRP preparations denatured and renatured in the presence of various cAMP concentrations. This finding suggests that there is no large-scale concerted motion (i.e., scissoring) of the CRP subunits in response to cAMP binding. In addition, we have identified three amino acid residues located along the CRP C alpha-helix that play a role in facilitating the conformation transition of the CRP hinge from that characteristic of apo-CRP to that characteristic of the CRP.cAMP complex. PMID- 17087498 TI - Characteristics of fibers formed by cytochrome c and induced by anionic phospholipids. AB - Recent publications described the formation of millimeter-length fibers by diverse lipid-binding proteins (e.g., histone H1, cytochrome c, indolicidin, and endostatin) when they are mixed with 80:20 phosphatidylcholine/phosphatidylserine vesicles. Further, these fibers displayed amyloid characteristics when stained with Congo Red. In the study presented here, we found by FTIR the amide I absorption band to reveal significant variation in fibers formed by cytochrome c, with some consisting of cytochrome c in a nativelike conformation and some exhibiting strong amyloid (beta-sheet) characteristics. Protein structure also varied from amyloid to nearly native within single fibers. Fibers were frequently blue or bluish and sometimes iridescent, likely due to interference of light in the fibers. The amyloid-type amide I band was observed for blue fibers only. AFM shows that fibers consist of smaller 3-4 nm diameter fibers with 10 nm lateral spacing. PMID- 17087499 TI - Structural perturbation of the carboxylate ligands to the manganese cluster upon Ca2+/Sr2+ exchange in the S-state cycle of photosynthetic oxygen evolution as studied by flash-induced FTIR difference spectroscopy. AB - A Ca(2+) ion is an indispensable element in the oxygen-evolving Mn cluster in photosystem II (PSII). To investigate the structural relevance of Ca(2+) to the Mn cluster, the effects of Sr(2+) substitution for Ca(2+) on the structures and reactions of ligands to the Mn cluster during the S-state cycle were investigated using flash-induced Fourier transform infrared (FTIR) difference spectroscopy. FTIR difference spectra representing the four S-state transitions, S(1) --> S(2), S(2) --> S(3), S(3) --> S(0), and S(0) --> S(1), were recorded by applying four consecutive flashes either to PSII core complexes from Thermosynechococcus elongatus or to PSII-enriched membranes from spinach. The spectra were also recorded using biosynthetically Sr(2+)-substituted PSII core complexes from T. elongatus and biochemically Sr(2+)-substituted PSII membranes from spinach. Several common spectral changes upon Sr(2+) substitution were observed in the COO(-) stretching region of the flash-induced spectra for both preparations, which were best expressed in Ca(2+)-minus-Sr(2+) double difference spectra. The significant intensity changes in the symmetric COO(-) peaks at approximately 1364 and approximately 1418 cm(-)(1) at the first flash were reversed as opposite intensity changes at the third flash, and the slight shift of the approximately 1446 cm(-)(1) peak at the second flash corresponded to the similar but opposite shift at the fourth flash. Analyses of these changes suggest that there are at least three carboxylate ligands whose structures are significantly perturbed by Ca(2+)/Sr(2+) exchange. They are (1) the carboxylate ligand having a bridging or unidentate structure in the S(2) and S(3) states and perturbed in the S(1) --> S(2) and S(3) --> S(0) transitions, (2) that with a chelating or bridging structure in the S(1) and S(0) states and perturbed also in the S(1) --> S(2) and S(3) --> S(0) transitions, and (3) that with a chelating structure in the S(3) and S(0) states and changes in the S(2) --> S(3) and S(0) --> S(1) transitions. Taking into account the recent FTIR studies using site-directed mutagenesis and/or isotope substitution [Chu et al. (2004) Biochemistry 43, 3152-3116; Kimura et al. (2005) J. Biol. Chem. 280, 2078-2083; Strickler et al. (2006) Biochemistry 45, 8801-8811], it was concluded that these carboxylate groups do not originate from either D1-Ala344 (C-terminus) or D1-Glu189, which are located near the Ca(2+) ion in the X-ray crystallographic model of the Mn cluster. It was thus proposed that if the X-ray model is correct, the above carboxylate groups sensitive to Sr(2+) substitution are ligands to the Mn ions strongly coupled to the Ca(2+) ion rather than direct ligands to Ca(2+). PMID- 17087500 TI - Redox modulation of cyclic electron flow around photosystem I in C3 plants. AB - We have investigated the occurrence of cyclic electron flow in intact spinach leaves. In particular, we have tested the hypothesis that cyclic flow requires the presence of supercomplexes in the thylakoid membrane or other strong associations between proteins. Using biochemical approaches, we found no evidence of the presence of supercomplexes related to cyclic electron flow, making previous structural explanations for the modulation of cyclic flow rather unlikely. On the other hand, we found that the fraction of photosystem I complexes engaged in cyclic flow could be modulated by changes in the redox state of the chloroplast stroma. Our findings support therefore a dynamic model for the occurrence of linear and cyclic electron flow in C3 plants, based on the competition between cytochrome b(6)f and FNR for electrons carried by ferredoxin. This would be ultimately regulated by the balance between the redox state of PSI acceptors and donors during photosynthesis, in a diffusing system. PMID- 17087501 TI - Identification of Rv3230c as the NADPH oxidoreductase of a two-protein DesA3 acyl CoA desaturase in Mycobacterium tuberculosis H37Rv. AB - DesA3 is a membrane-bound stearoyl-CoA Delta(9)-desaturase that produces oleic acid, a precursor of mycobacterial membrane phospholipids and triglycerides. The sequence of DesA3 is homologous with those of other membrane desaturases, including the presence of the eight-His motif proposed to bind the diiron center active site. This family of desaturases function as multicomponent complexes and thus require electron transfer proteins for efficient catalytic turnover. Here we present evidence that Rv3230c from Mycobacterium tuberculosis H37Rv is a biologically relevant electron transfer partner for DesA3 from the same pathogen. For these studies, Rv3230c was expressed as a partially soluble protein in Escherichia coli; recombinant DesA3 was expressed in Mycobacterium smegmatis as a catalytically active membrane protein. The addition of E. coli lysates containing Rv3230c to lysates of M. smegmatis expressing DesA3 gave strong conversion of [1 (14)C]-18:0-CoA to [1-(14)C]-cis-Delta(9)-18:1-CoA and of [1-(14)C]-16:0-CoA to [1-(14)C]-cis-Delta(9)-16:1-CoA. Both M. tuberculosis proteins were required for reconstitution of activity, as various combinations of control lysates lacking either Rv3230c or DesA3 gave minimal or no activity. Furthermore, the specificity of interaction between Rv3230c and DesA3 was implied by the inability of other related redox systems to substitute for Rv3230c. The reconstituted activity was dependent upon the presence of NADPH, could be saturated by increasing the amount of Rv3230c added, and was also sensitive to the salt concentration in the buffer. The results are consistent with the formation of a protein-protein complex, possibly with electrostatic character. This work defines a multiprotein, acyl-CoA desaturase complex from M. tuberculosis H37Rv to minimally consist of a soluble Rv3230c reductase and integral membrane DesA3 desaturase. Further implications of this finding relative to the properties of other multiprotein iron-enzyme complexes are discussed. PMID- 17087502 TI - Structure of a NADH-insensitive hexameric citrate synthase that resists acid inactivation. AB - Acetobacter aceti converts ethanol to acetic acid, and strains highly resistant to both are used to make vinegar. A. aceti survives acetic acid exposure by tolerating cytoplasmic acidification, which implies an unusual adaptation of cytoplasmic components to acidic conditions. A. aceti citrate synthase (AaCS), a hexameric type II citrate synthase, is required for acetic acid resistance and, therefore, would be expected to function at low pH. Recombinant AaCS has intrinsic acid stability that may be a consequence of strong selective pressure to function at low pH, and unexpectedly high thermal stability for a protein that has evolved to function at approximately 30 degrees C. The crystal structure of AaCS, complexed with oxaloacetate (OAA) and the inhibitor carboxymethyldethia coenzyme A (CMX), was determined to 1.85 A resolution using protein purified by a tandem affinity purification procedure. This is the first crystal structure of a "closed" type II CS, and its active site residues interact with OAA and CMX in the same manner observed in the corresponding type I chicken CS.OAA.CMX complex. While AaCS is not regulated by NADH, it retains many of the residues used by Escherichia coli CS (EcCS) for NADH binding. The surface of AaCS is abundantly decorated with basic side chains and has many fewer uncompensated acidic charges than EcCS; this constellation of charged residues is stable in varied pH environments and may be advantageous in the A. aceti cytoplasm. PMID- 17087503 TI - Crystal structure of an electron transfer complex between aromatic amine dehydrogenase and azurin from Alcaligenes faecalis. AB - The crystal structure of an electron transfer complex of aromatic amine dehydrogenase (AADH) and azurin is presented. Electrons are transferred from the tryptophan tryptophylquinone (TTQ) cofactor of AADH to the type I copper of the cupredoxin azurin. This structure is compared with the complex of the TTQ containing methylamine dehydrogenase (MADH) and the cupredoxin amicyanin. Despite significant similarities between the two quinoproteins and the two cupredoxins, each is specific for its respective partner and the ionic strength dependence and magnitude of the binding constant for each complex are quite different. The AADH azurin interface is largely hydrophobic, covering approximately 500 A(2) of surface on each molecule, with one direct hydrogen bond linking them. The closest distance from TTQ to copper is 12.6 A compared with a distance of 9.3 A in the MADH-amicyanin complex. When the MADH-amicyanin complex is aligned with the AADH azurin complex, the amicyanin lies on top of the azurin but is oriented quite differently. Although the copper atoms differ in position by approximately 4.7 A, the amicyanin bound to MADH appears to be rotated approximately 90 degrees from its aligned position with azurin. Comparison of the structures of the two complexes identifies features of the interface that dictate the specificity of the protein-protein interaction and determine the rate of interprotein electron transfer. PMID- 17087504 TI - Functional molecular mass of Escherichia coli K92 polysialyltransferase as determined by radiation target analysis. AB - The polysialyltransferase of Escherichia coli K92 catalyzes the transfer of sialic acid from CMP-sialic acid to a growing chain of polysialic acid at the nonreducing end. The enzyme encoded by the neuS gene is membrane-associated and has been suggested to be organized within a complex of several proteins encoded by the K92 gene cluster. Attempts to prepare a soluble active NeuS enzyme have been unsuccessful. Recent results suggest that de novo synthesis of polysialic acid requires coexpression of four genes from the cluster: neuS, neuE, kpsC, and kpsS. However, elongation of preexisting polysialic acid chains only requires expression of neuS. The molecular organization of the catalytic unit of bacterial polysialyltransferases has not been described. We used radiation inactivation to measure the size of the minimum functional unit catalyzing the polysialyltransferase chain extension and de novo reactions. Membranes harboring NeuS in the presence and absence of other products of the K92 gene cluster were exposed to high-energy electrons. The rate of loss of polysialyltransferase activity reveals the mass of the molecules essential for catalytic activity. We observed that the transfer of neuNAc from CMP-neuNAc to a polysialic acid acceptor is catalyzed by a complex with a target size larger than that of monomeric NeuS. The target size of the unit catalyzing the extension of existing polysialic acid chains does not differ significantly from the size of the unit catalyzing transfer of sialic acid to the endogenous acceptor. Parallel samples of membranes containing NeuS and a green fluorescent protein (GFP) chimera were compared by target analysis. The target size of this structural unit was estimated by analysis of the rate of decay of the GFP-NeuS chimera band migrating in the immunoblots. The target size of the structural unit is larger than expected for a monomer. The results of these experiments show that while the target size of the catalytic activity for K92 polysialyltransferase is larger than a monomer of NeuS, a large complex is not required for catalysis. PMID- 17087505 TI - Function of a conserved loop of the beta-domain, not involved in thiamin diphosphate binding, in catalysis and substrate activation in yeast pyruvate decarboxylase. AB - The X-ray crystal structure of pyruvamide-activated yeast pyruvate decarboxylase (YPDC) revealed a flexible loop spanning residues 290 to 304 on the beta-domain of the enzyme, not seen in the absence of pyruvamide, a substrate activator surrogate. Site-directed mutagenesis studies revealed that residues on the loop affect the activity, with some residues reducing k(cat)/K(m) by at least 1000 fold. In the pyruvamide-activated form, the loop located on the beta domain can transfer information to the active center thiamin diphosphate (ThDP) located at the interface of the alpha and gamma domains. The sigmoidal v(0)-[S] curve with wild-type YPDC attributed to substrate activation is modulated for most variants, but is not abolished. Pre-steady-state stopped-flow studies for product formation on these loop variants provided evidence for three enzyme conformations connected by two transitions, as already noted for the wild-type YPDC at pH 5.0 [Sergienko, E. A., and Jordan, F. (2002) Biochemistry 41, 3952-3967]. (1)H NMR analysis of the intermediate distribution resulting from acid quench [Tittmann et al. (2003) Biochemistry 42, 7885-7891] with all YPDC variants indicated that product release is rate limiting in the steady state. Apparently, the loop is not solely responsible for the substrate activation behavior, rather it may affect the behavior of residue C221 identified as the trigger for substrate activation. The most important function of the loop is to control the conformational equilibrium between the "open" and "closed" conformations of the enzyme identified in the pyruvamide-activated structure [Lu et al. (2000) Eur. J. Biochem. 267, 861-868]. PMID- 17087506 TI - Human cystathionine beta-synthase is a target for sumoylation. AB - Cystathionine beta-synthase (CBS) catalyzes the first irreversible step in the transsulfuration pathway and commits the toxic metabolite, homocysteine, to the synthesis of cysteine. Mutations in CBS are the most common cause of severe hereditary hyperhomocysteinemia. The molecular basis of the organ-specific pathologies associated with CBS deficiency is unknown as is the significance of the reported interaction between CBS and Huntingtin protein. In this study, we have used the yeast two-hybrid approach to screen for proteins that interact with CBS and have identified several components of the sumoylation pathway including Ubc9, PIAS1, PIAS3, Pc2, and RanBPM. We demonstrate that CBS is modified by the small ubiquitin-like modifier-1 protein (SUMO-I) under both in vitro and in vivo conditions. Deletion analysis of CBS indicates that the C-terminal regulatory domain is required for interaction with proteins in the sumoylation machinery. Sumoylated CBS is present in the nucleus where it is associated with the nuclear scaffold. The discovery that CBS is a target of sumoylation adds another layer to the complex regulation of this enzyme and reveals a previously unknown residence for this protein, i.e., in the nucleus. PMID- 17087507 TI - RecA dimers serve as a functional unit for assembly of active nucleoprotein filaments. AB - All RecA-like recombinase enzymes catalyze DNA strand exchange as elongated filaments on DNA. Despite numerous biochemical and structural studies of RecA and the related Rad51 and RadA proteins, the unit oligomer(s) responsible for nucleoprotein filament assembly and coordinated filament activity remains undefined. We have created a RecA fused dimer protein and show that it maintains in vivo DNA repair and LexA co-protease activities, as well as in vitro ATPase and DNA strand exchange activities. Our results support the idea that dimeric RecA is an important functional unit both for assembly of nucleoprotein filaments and for their coordinated activity during the catalysis of homologous recombination. PMID- 17087508 TI - Effect of DNA flanking sequence on charge transport in short DNA duplexes. AB - Several factors can influence charge transport (CT)-mediated DNA, such as sequence, distance, base stacking, base pair mismatch, conformation, tether length, etc. However, the DNA context effect or how flanking sequences influence redox active drugs in the DNA CT reaction and later in DNA enzymatic repair and synthesis is still not well understood. The set of seven DNA molecules in this study have been characterized well for the study of flanking sequence effects. These DNA duplexes are formed from self-complementary strands and contain the common central four-base sequence 5'-A-G-C-T-3', flanked on both sides by either (AT)(n) or (AA)(n) (n = 2, 3, or 4) or AA(AT)(2). UV-vis, fluorescence, UV melting, circular dichroism, and cyclic voltammetry experiments were used to study the flanking sequence effect on CT-mediated DNA by using daunomycin or adriamycin cross-linked with these seven DNA molecules. Our results showed that charge transport was related to the flanking sequence, DNA melting free energy, and ionic strength. For (AA)(n) or (AT)(n) species of the same length, (AA)(n) series were more stable and more efficient CT was observed through the (AA)(n) series. The same trend was observed for (AA)(n)() and (AT)(n) series at different ionic strengths, further supporting the idea that flanking sequence can result in different base stacking and modulate charge transport through these seven DNA molecules. PMID- 17087509 TI - Physical and structural basis for the strong interactions of the -ImPy- central pairing motif in the polyamide f-ImPyIm. AB - The polyamide f-ImPyIm has a higher affinity for its cognate DNA than either the parent analogue, distamycin A (10-fold), or the structural isomer, f-PyImIm (250 fold), has for its respective cognate DNA sequence. These findings have led to the formulation of a two-letter polyamide "language" in which the -ImPy- central pairings associate more strongly with Watson-Crick DNA than -PyPy-, -PyIm-, and ImIm-. Herein, we further characterize f-ImPyIm and f-PyImIm, and we report thermodynamic and structural differences between -ImPy- (f-ImPyIm) and -PyIm- (f PyImIm) central pairings. DNase I footprinting studies confirmed that f-ImPyIm is a stronger binder than distamycin A and f-PyImIm and that f-ImPyIm preferentially binds CGCG over multiple competing sequences. The difference in the binding of f ImPyIm and f-PyImIm to their cognate sequences was supported by the Na(+) dependent nature of DNA melting studies, in which significantly higher Na(+) concentrations were needed to match the ability of f-ImPyIm to stabilize CGCG with that of f-PyImIm stabilizing CCGG. The selectivity of f-ImPyIm beyond the four-base CGCG recognition site was tested by circular dichroism and isothermal titration microcalorimetry, which shows that f-ImPyIm has marginal selectivity for (A.T)CGCG(A.T) over (G.C)CGCG(G.C). In addition, changes adjacent to this 6 bp binding site do not affect f-ImPyIm affinity. Calorimetric studies revealed that binding of f-ImPyIm, f-PyImIm, and distamycin A to their respective hairpin cognate sequences is exothermic; however, changes in enthalpy, entropy, and heat capacity (DeltaC(p)) contribute differently to formation of the 2:1 complexes for each triamide. Experimental and theoretical determinations of DeltaC(p) for binding of f-ImPyIm to CGCG were in good agreement (-142 and -177 cal mol(-)(1) K(-)(1), respectively). (1)H NMR of f-ImPyIm and f-PyImIm complexed with their respective cognate DNAs confirmed positively cooperative formation of distinct 2:1 complexes. The NMR results also showed that these triamides bind in the DNA minor groove and that the oligonucleotide retains the B-form conformation. Using minimal distance restraints from the NMR experiments, molecular modeling and dynamics were used to illustrate the structural complementarity between f-ImPyIm and CGCG. Collectively, the NMR and ITC experiments show that formation of the 2:1 f-ImPyIm-CGCG complex achieves a structure more ordered and more thermodynamically favored than the structure of the 2:1 f-PyImIm-CCGG complex. PMID- 17087510 TI - Orientation and penetration depth of monolayer-bound p40phox-PX. AB - X-ray reflectivity was used to study the interaction of the PX domain of p40(phox) protein (p40(phox)-PX) with a Langmuir monolayer of a mixture of SOPC (1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine), SOPS (1-stearoyl-2-oleoyl-sn glycero-3-phosphoserine), and DPPtdIns(3)P (1,2-dipalmitoylphosphatidylinositol 3 phosphate) lipids supported on a buffered aqueous solution. The reflectivity is analyzed in terms of the known crystallographic structure of the p40(phox)-PX domain and a slab model that represents the lipid layer, yielding an electron density profile of the lipid layer and bound PX domains. This analysis determines the angular orientation and penetration depth of the p40(phox)-PX domain bound to the SOPC/SOPS/DPPtdIns(3)P monolayer. The best fit orientation is characterized by the following angles: theta = 30 +/- 10 degrees and phi = 140 +/- 30 degrees. These angles describe rotations, about axes in a coordinate system fixed to the domain, that are required to orient the domain with respect to the lipid layer at the interface. The protein penetrated into the lipid layer by 9 +/- 2 A, indicating that the protein penetrated into the headgroup region, but not deeply into the hydrocarbon region of the monolayer. In this analysis, polar Tyr(94) and hydrophobic Val(95) penetrated deepest into the lipid monolayer. The backbone of these residues was approximately 5 A above the headgroup-buffer interface, i.e., at the level of the SOPC/SOPS lipid phosphates. Positively charged Lys(92) and Lys(98) were also near the SOPC/SOPS lipid phosphates. This position of the protein allows for a favorable electrostatic contribution to binding. PMID- 17087511 TI - Hydrophobic loop dynamics and actin filament stability. AB - It has been postulated that the hydrophobic loop of actin (residues 262-274) swings out and inserts into the opposite strand in the filament, stabilizing the filament structure. Here, we analyzed the hydrophobic loop dynamics utilizing four mutants that have cysteine residues introduced at a single location along the yeast actin loop. Lateral, copper-catalyzed disulfide cross-linking of the mutant cysteine residues to the native C374 in the neighboring strand within the filament was fastest for S265C, followed by V266C, L267C, and then L269C. Site directed spin labeling (SDSL) studies revealed that C265 lies closest to C374 within the filament, followed by C266, C267, and then C269. These results are not predicted by the Holmes extended loop model of F-actin. Furthermore, we find that disulfide cross-linking destroys L267C and L269C filaments; only small filaments are observed via electron microscopy. Conversely, phalloidin protects the L267C and L269C filaments and inhibits their disulfide cross-linking. Combined, our data indicate that, in solution, the loop resides predominantly in a "parked" position within the filament but is able to dynamically populate other conformational states which stabilize or destabilize the filament. Such states may be exploited within a cell by filament-stabilizing and -destabilizing factors. PMID- 17087512 TI - Characterizing the residue level folding of the intrinsically unstructured IA3. AB - Residue level analysis of the folding of simple proteins may hold the key to understanding folding pathways and aid in structure prediction. IA(3), the endogenous inhibitor of yeast aspartic proteinase A (YPrA), is an unstructured protein in solution. Comparison of the 2D (15)N-HSQC spectra of IA(3) in water and in 23% 2,2,2-trifluoroethanol (TFE) shows that the individual residue cross peaks of IA(3) become more dispersed in the presence of TFE, indicating that the protein undergoes an unstructured to structured transition in the presence of TFE. This transition can be monitored by the movements of the cross peaks. Following the individual cross peaks, however, is complicated and does not establish whether a single transition occurs globally in the sequence. In this equilibrium study, we apply singular value decomposition (SVD) to elucidate both the main features of the TFE-driven transition and the residue-level deviations from the average behavior. This analysis has yielded a two-state folding description as well as specifics of NMR frequency shifts of individual residues, indicating that the N-terminus of IA(3) has a higher helical propensity than the C-terminus. Additionally, we discuss possible mechanisms for observed deviations from a two-state folding transition. When combined with a traditional biochemical understanding of interactions between individual residues, this approach leads to a better understanding of protein folding. PMID- 17087513 TI - Effects of disulfide bridges in domain I of Bacillus thuringiensis Cry1Aa delta endotoxin on ion-channel formation in biological membranes. AB - The delta-endotoxin family of toxic proteins represents the major component of the insecticidal capability of the bacterium Bacillus thuringiensis. Domain I of the toxins, which is largely alpha-helical, has been proposed to unfold at protein entry into the membrane of a target insect, following models known as the penknife and umbrella models. We extended the analysis of a previous work in which four disulfide bridges were constructed in domain I of the Cry1Aa delta endotoxin that putatively prevented unfolding during membrane partitioning. Using bioassays and voltage clamping of whole insect midgut instead of artificial lipid bilayers, it was found that, while toxicity and inhibition of the short-circuit current were reduced, only one of the disulfide bridges eliminated the activity of the toxins in the insect midgut membrane, and in that case, the loss of toxicity was due to the single amino acid substitution, R99C. It is proposed that at least alpha helices 4, 5, 6, and 7 and domain II partition in the midgut membranes of target insects, in support of an insertion model in which the whole protein translocates into the midgut membrane. PMID- 17087514 TI - Structural energetics and base-pair opening dynamics in sarcin-ricin domain RNA. AB - The sarcin-ricin domain is a universal element of the RNA from the large ribosomal subunit. The domain is part of the binding site for elongation factors and is specifically cleaved by the toxins alpha-sarcin and ricin. In this work, we have mapped the energetics and dynamics of individual structural motifs in a 29-mer RNA oligomer containing the sarcin-ricin domain. The stability of individual base pairs in the structure was characterized from measurements of the exchange rates of imino protons using nuclear magnetic resonance spectroscopy at 10 degrees C. The measurements also provided the rates of opening and closing for selected base pairs. The results reveal that the structural stabilization free energies in the sarcin-ricin domain are broadly distributed between 2.9 and 10.6 kcal/mol. One of the least stable sites in the structure is the noncanonical G-A base pair located next to the phosphodiester bond that is cleaved by alpha sarcin. The low stability of this base pair supports the proposal that cleavage by alpha-sarcin occurs by a base flipping mechanism. The opening dynamics of other base pairs is affected by elements of the structure such as the bulged-G motif and its cross-strand stacking. Participation in these motifs increases the lifetimes of the bases in an open, solvent-accessible conformation. PMID- 17087515 TI - Death-associated protein kinase phosphorylates mammalian ribosomal protein S6 and reduces protein synthesis. AB - Death-associated protein kinase (DAPK) is a pro-apoptotic, calcium/calmodulin regulated protein kinase that is a drug discovery target for neurodegenerative disorders. Despite the potential profound physiological role of DAPK in neuronal function and pathophysiology, the endogenous substrate(s) of this kinase and the mechanisms via which DAPK elicits its biological action remain largely unknown. We report here that the mammalian 40S ribosomal protein S6 is a DAPK substrate. Results from immunoprecipitation experiments are consistent with endogenous DAPK being associated with endogenous S6 in rat brain. When S6 is a component of the 40S ribosomal subunit complex, DAPK selectively phosphorylates it at serine 235, one of the five sites in S6 that are phosphorylated by the S6 kinase family of proteins. The amino acid sequence flanking serine 235 matches the established pattern for DAPK peptide and protein substrates. Kinetic analyses using purified 40S subunits revealed a K(m) value of 9 microM, consistent with S6 being a potential physiological substrate of DAPK. This enzyme-substrate relationship has functional significance. DAPK suppresses translation in rabbit reticulocyte lysate, and treatment of neuroblastoma cells with a stimulator of DAPK reduces protein synthesis. In both cases, suppression of translation correlates with increased phosphorylation of S6 at serine 235. These results demonstrate that DAPK is a S6 kinase and provide evidence for a novel role of DAPK in the regulation of translation. PMID- 17087516 TI - Metabolic processes account for the majority of the intracellular water in log phase Escherichia coli cells as revealed by hydrogen isotopes. AB - It is generally believed that water transport across biological membranes is essentially a near-instantaneous process, with water molecules diffusing directly across the membrane as well as through pores such as aquaporins. As a result of these processes by which water can equilibrate across a membrane, a common assumption is that intracellular water is isotopically indistinguishable from extracellular water. To test this assumption directly, we measured the hydrogen isotope ratio of intracellular water in Escherichia coli cells. Our results demonstrate that more than 50% of the intracellular water hydrogen atoms in log phase E. coli cells are isotopically distinct from the growth medium water and that these isotopically distinct hydrogen atoms are derived from metabolic processes. As expected, the (2)H/(1)H isotope ratio of intracellular water from log-phase cells showed an appreciably larger contribution from metabolic water than did intracellular water from stationary-phase cells (53 +/- 12 and 23 +/- 5%, respectively). The (2)H/(1)H isotope ratio of intracellular water was also monitored indirectly by measuring the isotope ratio of fatty acids, metabolites that are known to incorporate hydrogen atoms from water during biosynthesis. Significantly, the difference in the isotopic composition of intracellular water from log- to stationary-phase E. coli cells was reflected in the hydrogen isotope ratio of individual fatty acids harvested at the two different times, indicating that the isotope ratio of metabolites can be used as an indirect probe of metabolic activity. Together, these results demonstrate that contrary to the common assumption that intracellular water is isotopically identical to extracellular water, these two pools of water can actually be quite distinct. PMID- 17087517 TI - Analysis of the kinetic isotope effects on initial rates in transient kinetics. AB - A method was described recently for circumventing the difficulties in determining intrinsic kinetic isotope effects from eigenvalues obtained in transient kinetic experiments (Maniscalco, Tally, and Fisher (2004) Arch. Biochem. Biophys. 425, 165-172). The method, based on the isotope effects on initial rates of formation of intermediates, was validated by numerical simulation for only a few linear reaction sequences. A general analytical proof of the validity of the method is given in this work. The mathematical approach, using rate laws and L'Hopital's rule, allows more complex reaction schemes to be analyzed. Several are analyzed in this article, illustrating the broad applicability and possible complications in this approach for determining intrinsic isotope effects. Some possible applications are noted, with particular attention being paid to nonlinear reaction schemes, the effect of measuring signals rather than concentration, and the ability to distinguish stepwise from concerted reactions. PMID- 17087518 TI - Functional characterization of iron-substituted tristetraprolin-2D (TTP-2D, NUP475-2D): RNA binding affinity and selectivity. AB - The protein tristetraprolin (TTP, also known as NUP475 and TIS11) is a nonclassical zinc finger protein that is involved in regulating the inflammatory response. Specifically, TTP binds to AU-rich sequence elements located at the 3' untranslated region of cytokine mRNAs forming a complex that is degraded by the exosome. The nucleic acid binding region of TTP is comprised of two CysX(8)CysX(5)CysX(3)His domains that are activated in the presence of zinc. A two-domain construct of TTP (TTP-2D) has been cloned and overexpressed in E. coli. TTP-2D picks up visible red coloration from the expression media, unless it is expressed under iron-restricted conditions. The iron-binding properties of TTP 2D and the effect of iron substitution on RNA recognition have been investigated. Both Fe(II) and Fe(III) bind to TTP-2D and a full titration of Fe(III) with TTP 2D revealed that this metal ion binds with micromolar affinity. Upon reconstitution of TTP-2D with either Fe(II) or Fe(III), the protein recognizes a canonical RNA-binding sequence, UUUAUUUAUUU, with nanomolar affinity. Substitution of a single adenine or both adenines results in a decreased affinity of TTP-2D for the RNA molecule, demonstrating that both Fe(II)-TTP-2D and Fe(III) TTP-2D selectively recognize a physiologically relevant RNA sequence. The relative affinities of Fe(II)-TTP-2D and Fe(III)-TTP-2D for the series of RNA sequences mirror those observed for Zn(II)-TTP-2D and suggest that iron is a viable substitute for zinc in this protein. PMID- 17087519 TI - X-ray absorption spectroscopy of the zinc-binding sites in the class B2 metallo beta-lactamase ImiS from Aeromonas veronii bv. sobria. AB - X-ray absorption spectroscopy was used to investigate the metal-binding sites of ImiS from Aeromonas veronii bv. sobria in catalytically active (1-Zn), product inhibited (1-Zn plus imipenem), and inactive (2-Zn) forms. The first equivalent of zinc(II) was found to bind to the consensus Zn(2) site. The reaction of 1-Zn ImiS with imipenem leads to a product-bound species, coordinated to Zn via a carboxylate group. The inhibitory binding site of ImiS was examined by a comparison of wild-type ImiS with 1 and 2 equiv of bound zinc. 2-Zn ImiS extended X-ray absorption fine structure data support a binding site that is distant from the active site and contains both one sulfur donor and one histidine ligand. On the basis of the amino acid sequence of ImiS and the crystal structure of CphA [Garau et al. (2005) J. Mol. Biol. 345, 785-795], we propose that the inhibitory binding site is formed by M146, found on the B2-distinct alpha3 helix, and H118, a canonical Zn(1) ligand, proposed to help activate the nucleophilic water. The mutation of M146 to isoleucine abolishes metal inhibition. This is the first characterization of ImiS with the native metal Zn and establishes, for the first time, the location of the inhibitory metal site. PMID- 17087520 TI - In vitro biosynthesis of UDP-N,N'-diacetylbacillosamine by enzymes of the Campylobacter jejuni general protein glycosylation system. AB - In Campylobacter jejuni 2,4-diacetamido-2,4,6-trideoxy-alpha-d-glucopyranose, termed N,N'-diacetylbacillosamine (Bac2,4diNAc), is the first carbohydrate in the glycoprotein N-linked heptasaccharide. With uridine diphosphate-N acetylglucosamine (UDP-GlcNAc) as a starting point, two enzymes of the general protein glycosylation (Pgl) pathway in C. jejuni (PglF and PglE) have recently been shown to modify this sugar nucleotide to form UDP-2-acetamido-4-amino-2,4,6 trideoxy-alpha-d-glycopyranose (UDP-4-amino-sugar) [Schoenhofen, I. C., et al. (2006) J. Biol. Chem. 281, 723-732]. PglD has been proposed to catalyze the final step in N,N'-diacetylbacillosamine synthesis by N-acetylation of the UDP-4-amino sugar at the C4 position. We have cloned, overexpressed, and purified PglD from the pgl locus of C. jejuni NCTC 11168 and identified it as the acetyltransferase that modifies the UDP-4-amino-sugar to form UDP-N,N'-diacetylbacillosamine, utilizing acetyl-coenzyme A as the acetyl group donor. The UDP-N,N' diacetylbacillosamine product was purified from the reaction by reverse phase C18 HPLC and the structure determined by NMR analysis. Additionally, the full-length PglF was overexpressed and purified in the presence of detergent as a GST fusion protein, allowing for derivation of kinetic parameters. We found that the UDP-4 amino-sugar was readily synthesized from UDP-GlcNAc in a coupled reaction using PglF and PglE. We also demonstrate the in vitro biosynthesis of the complete heptasaccharide lipid-linked donor by coupling the action of eight enzymes (PglF, PglE, PglD, PglC, PglA, PglJ, PglH, and PglI) in the Pgl pathway in a single reaction vessel. PMID- 17087521 TI - Thermal stabilities of brain spectrin and the constituent repeats of subunits. AB - The different genes that encode mammalian spectrins give rise to proteins differing in their apparent stiffness. To explore this, we have compared the thermal stabilities of the structural repeats of brain spectrin subunits (alphaII and betaII) with those of erythrocyte spectrin (alphaI and betaI). The unfolding transition midpoints (T(m)) of the 36 alphaII- and betaII-spectrin repeats extend between 24 and 82 degrees C, with an average higher by some 10 degrees C than that of the alphaI- and betaI-spectrin repeats. This difference is reflected in the T(m) values of the intact brain and erythrocyte spectrins. Two of three tandem-repeat constructs from brain spectrin exhibited strong cooperative coupling, with elevation of the T(m) of the less stable partner corresponding to coupling free energies of approximately -4.4 and -3.5 kcal/mol. The third tandem repeat construct, by contrast, exhibited negligible cooperativity. Tandem-repeat mutants, in which a part of the "linker" helix that connects the two domains was replaced with a corresponding helical segment from erythroid spectrin, showed only minor perturbation of the thermal melting profiles, without breakdown of cooperativity. Thus, the linker regions, which tolerate few point mutations without loss of cooperative function, have evidently evolved to permit conformational coupling in specified regions. The greater structural stability of the repeats in alphaII- and betaII-spectrin may account, at least in part, for the higher rigidity of brain compared to erythrocyte spectrin. PMID- 17087522 TI - The influence of Mayan education on middle school students in Guatemala. AB - The purpose of this study was to examine the influence of Mayan education on the academic achievement of Indian and Ladino middle school students (N = 353) in Guatemala. This study also examined changes in ethnic identity achievement and the effects of changes in ethnic identity achievement on gains in self-esteem and other-group attitudes. Superior gains in academic skills for both Ladino and Indian students attending Mayan schools were found. The results also suggested that those students who increased their ethnic identity scores during their first year of middle school also increased their other-group attitudes. These results are discussed in terms of the benefits of Mayan education and ethnic identity achievement for both Indian and Ladino students. PMID- 17087524 TI - A qualitative examination of the relationships that serve a mentoring function for Mexican American older adolescents. AB - This exploratory study was an in-depth examination of Mexican American adolescents' relationships with nonparental adults. Qualitative interviews with 10 Mexican American adolescents revealed 23 nonparental adults who served a mentoring function in their lives. Six of these nonparental adults were also interviewed. Data analyses were conducted using a grounded theory approach so that the relationships were described in participants' words and experiences. The nonparental adults identified by adolescents included siblings, extended family members, older peers, and institutional figures. The support provided took many different forms, from emotional to informational/experiential support, to modeling behavior, for example. Further, adolescents were supported in eight different areas of their lives. Participants also discussed the perceived benefits of these relationships for adolescents. Future research directions and implications for youth programming are discussed. PMID- 17087525 TI - Testing of an orthogonal measure of cultural identification with adult mission Indians. AB - The Orthogonal Cultural Identification Scale (OCIS; Oetting & Beauvais, 1990-91) has been validated among Native American youth, but not adults. The present study sought to test the reliability (internal consistency) and validity (construct and factorial) of the OCIS among an adult Native American sample consisting of 389 Mission Indians (61% female). Participants were recruited from reservations using a venue sampling strategy. The OCIS was completed as part of a self-assessment packet of questionnaires. Internal consistency for OCIS subscale scores ranged from 0.76 to 0.91. Both concurrent and discriminant validity were demonstrated. Confirmatory factor analysis revealed 2 factors: (1) Anglo American Identification and (2) Native American Identification. These results indicate the OCIS is a reliable and valid instrument for use with adult Native Americans. PMID- 17087526 TI - Gender differences in early risk factors for adolescent depression among low income urban children. AB - One component of a model by Nolen-Hoeksema and Girgus, who propose that risk factors for adolescent depression are more common in girls than in boys during childhood, was tested with 85 low-income, urban, African American and Latino kindergarten through fourth grade children who completed inventories of depression, stress, attributional style, gender role, and body image. Endorsing two of three predicted risk factors, girls reported slightly poorer body image and identified more strongly with a feminine gender role. Boys, however, reported a more negative attributional style. Feminine gender role was not associated with body image or negative attributional style. The applicability of the proposed model to a low-income, ethnic minority, urban population is discussed. PMID- 17087527 TI - Discrimination and exiting homelessness among homeless adolescents. AB - This article examines how newly homeless adolescents' discrimination experiences were associated with exiting homelessness after 6 months. A sample of 262 homeless adolescents, aged 12 to 20 years, were recruited and followed longitudinally (6-month retention rate = 88%). Discrimination was related to being gay, lesbian, or bisexual (LGB). Discrimination from family was related to exiting homelessness. Other than those who were LGB, adolescents who reported discrimination from their families were more likely to exit homelessness than adolescents who did not report such discrimination. Suggestions for future research include focusing on the experiences of LGB homeless adolescents, the role of families in the lives of homeless adolescents, and other aspects of discrimination, including salience, frequency, intensity, and duration. PMID- 17087528 TI - Relationship between physical appearance, sense of belonging and exclusion, and racial/ethnic self-identification among multiracial Japanese European Americans. AB - In this study the authors explored the relation of physical appearance, perception of group belonging, and perception of group exclusion to racial/ethnic identity in multiracial Japanese European Americans. Results indicate that physical appearance and social variables of sense of belonging and exclusion related to one monoracial racial/ethnic group significantly predicted self identity with the corresponding monoracial group. There was also a significant relationship between Japanese American identity and multiracial appearance and social variables. Feelings of exclusion were shown to be the primary influence on all three racial/ethnic identities. PMID- 17087529 TI - Latina college students' sexual health beliefs about human papillomavirus infection. AB - To examine Latina college students' sexual health beliefs about human papillomavirus (HPV), 16 self-identified Latina college students from a large Midwestern university participated in three focus groups. The authors found that participants' knowledge of HPV and cervical cancer was extremely low. Women did not obtain Pap screenings consistently, nor did they use condoms in a consistent manner. They perceived that cultural norms that support silence about sexual topics hinder their knowledge acquisition about sexual issues. Participants also recognized that their insufficient knowledge could jeopardize their health and expressed a willingness to actively stop this intergenerational cycle of silence within their families. Findings highlight the importance of incorporating cultural considerations when designing research and educational programs with Latina college students. PMID- 17087530 TI - Perceived family conflict, parental attachment, and depression in African American female adolescents. AB - The purpose of this study was to test a hypothesized model that explored the degree to which parental attachment mediated the relationship between perceived family conflict and depression in a sample of 283 African American female adolescents. Results revealed that perceived family conflict had both a direct and an indirect effect on depression, with parental attachment mediating 28% of the effect of perceived family conflict on depression. Implications of the findings are discussed. PMID- 17087531 TI - Inter- and intra-group variability in perceptions of behavior among Asian Americans and European Americans. AB - Between-group and within-group differences in perceptions of behavior were examined among 74 Asian Americans and 111 European Americans. Participants rated videotaped interactions of Asian American mothers and daughters. Asian Americans, and a more homogeneous sample of Chinese Americans, perceived less maternal control and more reciprocity than did European Americans. Intra-cultural variations in perceptions were also evident, as within-group analyses revealed differences in perceptions based on the generational status of Asian Americans and the degree of multicultural experience of European Americans. Overall, findings suggest that perceptions of behavior are shaped as much by within-group differences in familiarity and experience with the target culture as by between group differences in ethnicity. Implications for theories of child socialization, multicultural counseling, and observational research are discussed. PMID- 17087532 TI - Implications of adolescents' acculturation strategies for personal and collective self-esteem. AB - Berry, Trimble, and Olmedo's (1986) acculturation model was used to investigate the relationship among adolescents' acculturation strategies, personal self esteem, and collective self-esteem. Using data from 427 high school students, factor analysis results distinguished Collective Self-esteem Scale constructs (Luhtanen & Crocker, 1992) from both ethnic identity and outgroup orientation subscales of the Multigroup Ethnic Identity Measure (Phinney, 1992). Subsequent results showed that: 1) both acculturation dimensions were correlated with personal and collective self-esteems, 2) integrationists shared similar levels of personal and collective self-esteems with assimilationists and/or separationists, and 3) marginalizationists generally had the lowest levels of personal and collective self-esteems. Implications are drawn for understanding acculturation among adolescents and for the utility of group-level measures of self-esteem. PMID- 17087533 TI - Developmental patterns of African American and Caucasian adolescents' alcohol use. AB - This study examined the developmental alcohol use trajectories of Caucasian and African American adolescents. Participants were 1,358 adolescents (77.6% Caucasian). Results suggest that not only do patterns of alcohol use differ for Caucasian and African American adolescents, but that there is substantial within group variation as well. Results suggest that using a within-groups developmental approach is one promising way to identify subgroups of adolescents at greatest risk for substance use. PMID- 17087535 TI - The effects of antipoverty programs on children's cumulative level of poverty related risk. AB - The authors examined the effects of antipoverty programs on children's cumulative poverty-related risk and the relationship between cumulative poverty-related risk and child outcomes among low-income families. Samples included 419 children ages 3-10 years in the New Hope program and 759 children ages 2-9 years in the Minnesota Family Investment Program (MFIP), which tested 2 program approaches. Nine poverty-related risks made up the measure of cumulative risk. Both MFIP program approaches reduced cumulative poverty-related risk. New Hope reduced cumulative poverty-related risk among long-term welfare recipients. In both New Hope and MFIP, significant linear relationships between cumulative poverty related risk and parent-reported behavior problems and school achievement were found. Cumulative poverty-related risk partially mediated the impacts of the MFIP programs on children's behavior problems. Among long-term welfare recipients, cumulative poverty-related risk partially mediated New Hope's impact on parent reported school achievement. PMID- 17087536 TI - Infants flexibly use different dimensions to categorize objects. AB - A sequential-touching task was used to investigate whether 14-month-old infants can rapidly change how they categorize a set of objects, recognizing new groupings of objects they had previously categorized in a different way. When presented with a collection of objects that could be categorized by shape (balls vs. blocks) or material (soft vs. hard), infants who showed stable performance on a superordinate-level categorization task or who had larger receptive vocabularies exhibited flexible categorization; they categorized the objects by material as well as by shape. Infants who rarely responded to the superordinate level categorization task or who had smaller receptive vocabularies, in contrast, categorized primarily by shape. Thus, flexible categorization is related to development in other cognitive domains. PMID- 17087538 TI - The developmental origins of cognitive vulnerability to depression: temperament, parenting, and negative life events in childhood as contributors to negative cognitive style. AB - Cognitive models of depression have been well supported with adults, but the developmental origins of cognitive vulnerability are not well understood. The authors hypothesized that temperament, parenting, and negative life events in childhood would contribute to the development of cognitive style, with withdrawal negativity and negative parental feedback moderating the effects of negative life events to predict more depressogenic cognitive styles. These constructs were assessed in 289 children and their parents followed longitudinally from infancy to 5th grade; a subsample (n = 120) also participated in a behavioral task in which maternal feedback to child failure was observed. Results indicated that greater withdrawal negativity in interaction with negative life events was associated with more negative cognitive styles. Self-reported maternal anger expression and observed negative maternal feedback to child's failure significantly interacted with child's negative events to predict greater cognitive vulnerability. There was little evidence of paternal parenting predicting child negative cognitive style. PMID- 17087539 TI - In search of shared and nonshared environmental factors in security of attachment: a behavior-genetic study of the association between sensitivity and attachment security. AB - The current article presents results from a twin study of genetic and environmental components of maternal sensitivity and infant attachment and their association. The sample consisted of 136 twin pairs from 2 sites: Leiden, the Netherlands, and London, UK. Maternal sensitivity was assessed in the home at 9 10 months, and infant attachment security was observed in the laboratory at 12 months. The study yielded little evidence that genetic factors are involved in variations between twins in maternal sensitivity ratings but did find that shared variance in maternal sensitivity was able to account for some of the similarity between twins in attachment security. Weak nonshared associations between sensitivity and attachment appeared to suppress the magnitude of the correlation between attachment and sensitivity in twin children. The results could indicate that the attachment security of one twin may depend on the relationship the parent has with the other twin. The results are brought to bear on the validity of attachment theory as a theory of primarily shared environmental effects in children's development and the continuing challenge posed to attachment theory by within-family differences in socioemotional processes. PMID- 17087540 TI - Predictors of paternal involvement for resident and nonresident low-income fathers. AB - In a sample of low-income families (N = 239), structural equation models assessed predictors of fathers' involvement with preschool-aged children in instrumental, behavioral, and emotional realms. Results suggest that parental conflict has a strong negative relation with father involvement. Fathers' human capital characteristics, healthy psychosocial functioning, and past stability in family relationships all predicted greater father involvement directly and/or indirectly through parental conflict. Numerous differences emerged in the predictive models between resident and nonresident fathers, although few differences were statistically significant. Results suggest that policy efforts aimed at enhancing fathers' responsible parenting should focus both on increasing fathers' human and social capital and on supporting positive family processes. PMID- 17087541 TI - Links between social network closure and child well-being: the organizing role of friendship context. AB - Third grade children (N = 404) and their mothers completed questionnaires and participated in interviews designed to identify children's friendships across multiple contexts, determine levels of social network closure for these friendships, and assess child well-being. Cluster analyses revealed distinct patterns in the contexts in which children's friendships were maintained. Closure was highest for children whose friendship clusters heavily represented relatives as friends and lowest when friends were from schools and the broader community. Intermediate levels of closure were observed for the clusters of neighborhood friends and friends from church and school. Both friendship cluster and, to some extent, ethnicity moderated associations between closure and indicators of well being. PMID- 17087542 TI - Visual processing and infant ocular Latencies in the overlap paradigm. AB - Young infants have repeatedly been shown to be slower than older infants to shift fixation from a midline stimulus to a peripheral stimulus. This is generally thought to reflect maturation of the neural substrates that mediate the disengagement of attention, but this developmental difference may also be attributable to young infants' slower processing of the midline stimulus. This possibility was tested with 3- and 7-month-old infants in 2 experiments in which the degree of familiarity of the midline stimulus was manipulated across repeated trials. The results of these experiments demonstrated that the processing of midline content does affect infants' ocular latencies to a peripheral stimulus but that developmental differences in such processing do not account for developmental differences in disengagement seen across the 1st year. PMID- 17087543 TI - Contextual basis of maternal perceptions of infant temperament. AB - To elucidate the differential saliency of infant emotions to mothers across interactive contexts, the authors examined the moderating role of observed infant affect during interactions with mother in the relation between maternal and laboratory-based ratings of infant temperament. Fifty-nine developmentally healthy 9-month-old infants were judged for degree of infant positive, infant negative, and mother-infant mutually positive affect during the course of object focused and routine home-based activities with mother. Mothers completed the Infant Behavior Questionnaire (M. K. Rothbart, 1981), and infants underwent the Laboratory Temperament Assessment Battery (H. H. Goldsmith & M. K. Rothbart, 1999). Results revealed that maternal and observer ratings of infant negativity converged when infants manifested high degrees of negative affect during routine home-based activities. Maternal and observer ratings of infant positivity converged when infants experienced low mutually positive affect during play. These findings support the hypothesis that maternal perceptions are based on mothers' experiences with their infants but that the salience of infant temperamental characteristics to mothers varies across emotion and interactive context. PMID- 17087544 TI - Life-span development of visual working memory: when is feature binding difficult? AB - We asked whether the ability to keep in working memory the binding between a visual object and its spatial location changes with development across the life span more than memory for item information. Paired arrays of colored squares were identical or differed in the color of one square, and in the latter case, the changed color was unique on that trial (item change) or was duplicated elsewhere in the array (color-location binding change). Children (8-10 and 11-12 years old) and older adults (65-85 years old) showed deficits relative to young adults. These were only partly simulated by dividing attention in young adults. The older adults had an additional deficiency, specifically in binding information, which was evident only when item- and binding-change trials were mixed together. In that situation, the older adults often overlooked the more subtle, binding-type changes. Some working memory processes related to binding undergo life-span development in an inverted-U shape, whereas other, bias- and salience-related processes that influence the use of binding information seem to develop monotonically. PMID- 17087545 TI - Blickets and babies: the development of causal reasoning in toddlers and infants. AB - Previous research has suggested that preschoolers possess a cognitive system that allows them to construct an abstract, coherent representation of causal relations among events. Such a system lets children reason retrospectively when they observe ambiguous data in a rational manner (e.g., D. M. Sobel, J. B. Tenenbaum, & A. Gopnik, 2004). However, there is little evidence that demonstrates whether younger children possess similar inferential abilities. In Experiment 1, the authors extended previous findings with older children to examine 19- and 24 month-olds' causal inferences. Twenty-four-month-olds' inferences were similar to those of preschoolers, but younger children lacked the ability to make retrospective causal inferences, perhaps because of performance limitations. In Experiment 2, the authors designed an eye-tracking paradigm to test younger participants that eliminated various manual search demands. Eight-month-olds' anticipatory eye movements, in response to retrospective data, revealed inferences similar to those of 24-month-olds in Experiment 1 and preschoolers in previous research. These data are discussed in terms of associative reasoning and causal inference. PMID- 17087546 TI - Popularity, social acceptance, and aggression in adolescent peer groups: links with academic performance and school attendance. AB - This article reports a short-term longitudinal study focusing on popularity and social acceptance as predictors of academic engagement for a sample of 342 adolescents (approximate average age of 14). These youths were followed for 4 consecutive semesters. Popularity, social acceptance, and aggression were assessed with a peer nomination inventory, and data on academic engagement were obtained from school records. For adolescents who were highly aggressive, increases in popularity were associated with increases in unexplained absences and decreases in grade point average. Conversely, changes in social acceptance were not predictive of changes in grade point average or unexplained absences. These results highlight the importance of multidimensional conceptualizations of social standing for research on school adjustment during adolescence and emphasize the potential risks associated with popularity. PMID- 17087547 TI - Predicting delayed letter knowledge development and its relation to grade 1 reading achievement among children with and without familial risk for dyslexia. AB - The authors examined the developmental trajectories of children's early letter knowledge in relation to measures spanning and encompassing their prior language related and cognitive measures and environmental factors and their subsequent Grade 1 reading achievement. Letter knowledge was assessed longitudinally at ages 4.5, 5.0, 5.5, and 6.5 years; earlier language skills and environmental factors were assessed at ages 3.5 and 4.5 years; and reading achievement was assessed at the beginning and end of Grade 1. The analyses were conducted on a longitudinal data set involving children with and without familial risk for dyslexia. Emerging from the trajectory analysis of letter knowledge were 3 separate clusters: delayed (n = 63), linearly growing (n = 73), and precocious (n = 51). The members of the delayed cluster were predominantly children with familial risk for dyslexia, and the members of the precocious cluster were predominantly control group children. Phonological sensitivity, phonological memory, and rapid naming skills predicted delayed letter knowledge. Environmental predictors included level of maternal education and the amount of letter name teaching. Familial risk for dyslexia made a significant contribution to the predictive relations. Membership in the delayed cluster predicted poor reading performance at Grade 1. PMID- 17087548 TI - Social development from infancy to adolescence: longitudinal and concurrent factors in an adoption sample. AB - In the present longitudinal study, early adopted children (N = 160) were followed from infancy to adolescence to assess the influence of previous and concurrent factors on the children's social development. This study allowed for more conclusive evidence of the influence of early and concurrent rearing experiences and temperament on adolescents' social development, independent of shared genetic factors between children and parents. Results showed that social development and temperament were stable over time and that both previous and current parental sensitivity were important in predicting social development in adolescence. Quality of the early parent-child relationship was indirectly associated with social development in adolescence through the influence on social development in middle childhood. Maternal sensitivity in middle childhood and in adolescence partly buffered the negative effects of difficult temperament on social development in adolescence. Adaptation emerged as the product of both developmental history and current circumstances. PMID- 17087549 TI - Economic disparities in middle childhood development: does income matter? AB - A large literature has documented the influence of family economic resources on child development, yet income's effects in middle childhood have been understudied. Using data from the National Longitudinal Survey of Youth (N = 3,551), the author examined the influence of family income in early and middle childhood on academic skills and behavior problems during middle childhood. Early childhood income had enduring effects on children's behavior problems and academic skills in middle childhood. Middle childhood income did not influence academic skills but did affect the development of behavior problems during middle childhood. Children from low-income households were particularly sensitive to the effects of family income. The quality of home environment during early and middle childhood explained a portion of the effects of income on academic skills and behavior problems. PMID- 17087550 TI - Adult social behavioral effects of heavy adolescent marijuana use among African Americans. AB - The authors examined the effects of heavy adolescent marijuana use on employment, marriage, and family formation and tested both dropping out of high school and adult marijuana use as potential mediators of these associations among a community sample of African Americans followed longitudinally from age 6 to age 32-33. They used propensity score matching to reduce selection bias when estimating the effects of heavy adolescent marijuana use. Logistic regression results on the sample matched on sex, and early demographic and behavioral variables showed that adolescent marijuana use has adult social behavioral consequences: Use of marijuana 20 times or more during adolescence was associated with being unemployed and unmarried in young adulthood and having children outside of marriage for both males and females. Dropping out of high school and more frequent adult marijuana use seem to be important parts of the pathway from adolescent marijuana use to negative life outcomes. PMID- 17087551 TI - Class climate moderates peer relations and emotional adjustment in children with an early history of anxious solitude: a Child X Environment model. AB - Classroom emotional climate was hypothesized to moderate psychosocial adjustment in 1st grade for children with an early childhood history of anxious solitude. Participants were 1,364 children in the National Institute of Child Health and Human Development (NICHD) Study of Early Child Care and their mothers, child-care providers, and teachers. As anticipated, children with an early childhood history of anxious solitude were more rejected, poorly accepted (boys), and victimized (girls) by peers and demonstrated more depressive symptoms (girls) in 1st-grade classrooms with negative observed emotional climate. Results support a Child x Environment model of children's social and emotional adjustment. PMID- 17087552 TI - Developmental invariance in distinctiveness effects in memory. AB - Improvements in 5- and 7-year-olds' acquisition and retention of related concept pairings were examined when additional similarities and differences between pair members were provided. Using a standard paired-associate learning paradigm, children learned 18 related picture pairs; some of the children either were given or produced additional similarities or differences between pair members at the time of learning. Three weeks after learning was complete, children attempted to recall the pairs. Using a model to determine the storage and retrieval loci of these effects, the results showed that (a) all children benefited from self generated elaborations, regardless of whether these were similarities or differences, and these benefits were storage related, and (b) difference elaborations improved children's retention regardless of whether they were self- or experimenter-generated, and these effects were primarily retrieval based. These results are consistent with theories that (a) view retrieval as the locus of distinctiveness effects and (b) view storage as the locus of self-generated memory improvements. PMID- 17087553 TI - Cognitive engagement and story comprehension in typically developing children and children with ADHD from preschool through elementary school. AB - The present study examined children's cognitive engagement with television as a function of the continuity of central or incidental content and whether this varied with age and clinical status. In Experiment 1, 9- to 11-year-old children's response times on a secondary task were slower the later a probe occurred in a sequence of central events, and response times predicted recall. Experiment 2 extended these results to 6- to 8-year-old children. Experiment 3 revealed that children with attention-deficit/hyperactivity disorder (ADHD) failed to show the pattern consistently observed for comparison children. The results support the hypothesis that typically developing children build a representation during viewing that reflects the causal structure of the televised story but that this skill is deficient in 4- to 9-year-old children with ADHD. PMID- 17087554 TI - Co-occurring delinquency and depressive symptoms of adolescent boys and girls: a dual trajectory modeling approach. AB - Co-occurring trajectories of delinquent behavior and depressive symptoms and their correlates were examined in a longitudinal sample of 985 middle-adolescent boys and girls (mean age = 15.54 years at Time 1). Dual trajectory analysis was used to identify the co-occurring trajectories. For boys (n = 472), 4 delinquency and 4 depression trajectory groups were found. For girls (n = 513), 3 delinquency and 3 depression trajectory groups were identified. The linkage between co occurring trajectories was higher for girls than for boys. Stressful life events and childhood precursors of the outcomes predicted trajectory group membership for both genders fairly consistently. Findings suggest heterogeneity in developmental courses of delinquent behavior and depressive symptoms across adolescent boys and girls. PMID- 17087555 TI - Learning by selection: visual search and object perception in young infants. AB - The authors examined how visual selection mechanisms may relate to developing cognitive functions in infancy. Twenty-two 3-month-old infants were tested in 2 tasks on the same day: perceptual completion and visual search. In the perceptual completion task, infants were habituated to a partly occluded moving rod and subsequently presented with unoccluded broken and complete rod test stimuli. In the visual search task, infants viewed displays in which single targets of varying levels of salience were cast among homogeneous static vertical distractors. Infants whose posthabituation preference indicated unity perception in the completion task provided evidence of a functional visual selective attention mechanism in the search task. The authors discuss the implications of the efficiency of attentional mechanisms for information processing and learning. PMID- 17087556 TI - Dimensions of family connectedness among adolescents with Mexican, Chinese, and European backgrounds. AB - Multiple dimensions of adolescents' connectedness with their families were investigated among 489 9th-grade students (M = 14.86 years) from families with Mexican, Chinese, and European backgrounds. Participants reported on various aspects of their family relationships and completed diary checklists of daily behaviors for a 2-week period. Adolescents from European backgrounds reported levels of family identification and dyadic closeness with parents similar to or greater than those reported by their peers. For adolescents from Mexican and Chinese backgrounds, particularly those from immigrant families, family connectedness included a stronger emphasis on family obligation and assistance. The extent to which family demographic variables, including parental level of education and residence in a single-parent family, accounted for group differences was examined. PMID- 17087558 TI - The importance of gesture in children's spatial reasoning. AB - On average, men outperform women on mental rotation tasks. Even boys as young as 4 1/2 perform better than girls on simplified spatial transformation tasks. The goal of our study was to explore ways of improving 5-year-olds' performance on a spatial transformation task and to examine the strategies children use to solve this task. We found that boys performed better than girls before training and that both boys and girls improved with training, whether they were given explicit instruction or just practice. Regardless of training condition, the more children gestured about moving the pieces when asked to explain how they solved the spatial transformation task, the better they performed on the task, with boys gesturing about movement significantly more (and performing better) than girls. Gesture thus provides useful information about children's spatial strategies, raising the possibility that gesture training may be particularly effective in improving children's mental rotation skills. PMID- 17087559 TI - Identity and spirituality: a psychosocial exploration of the sense of spiritual self. AB - The authors examined the structure and content of adults' sense of spiritual identity by analyzing semistructured interviews with 13 spiritually devout men and 15 devout women, ages 22 to 72. Individuals' responses to the Role-Related Identity Interview (G. T. Sorell, M. J. Montgomery, & N. A. Busch-Rossnagel, 1997b) were content analyzed and rated on the role-related spiritual identity dimensions of role salience and flexibility. Individuals were categorized as spiritually foreclosed, achieved, or in moratorium, on the basis of their motivational, affective, self-evaluative, and behavioral investments in spiritually defined roles and their reflectiveness about and behavioral changes in role-related spiritual identity. Similarities and differences within and between spiritual identity status groups were observed, suggesting a variety of ways that spiritual identity provides a sense of continuity as well as a domain for adult developmental change. PMID- 17087560 TI - Effects of discrete emotions on young children's ability to discern fantasy and reality. AB - This study examined 3- to 5-year-olds' (N = 128; 54% girls) ability to discriminate emotional fantasy and reality. Children viewed images depicting fantastic or real events that elicited several emotions, reported whether each event could occur, and rated their emotional reaction to the image. Children were also administered the Play Behavior Questionnaire and Pretend Action Tasks to assess play behaviors. Findings revealed age-related improvements in performance and biases in judgment based on the emotion depicted. Children reported that happy fantastic events could occur significantly more often than frightening and angry fantastic events and that happy real events could occur significantly more often than frightening and angry real events. Children's emotional reactions to the images but not play behaviors were significantly related to their fantasy reality distinctions. Implications for the relation between emotions and children's fantasy-reality distinctions are discussed. PMID- 17087561 TI - Differential parent-child relationships and adolescent externalizing symptoms: cross-lagged analyses within a monozygotic twin differences design. AB - Research has indicated that differential parental treatment is linked to differences in externalizing symptomology (EXT) across siblings, even those siblings who are genetically identical. However, the direction of causation and longitudinal significance of this relationship remains unclear. Thus, in the present study, the authors examined 486 monozygotic twin pairs, assessed at ages 11, 14, and 17 years, within a cross-lagged twin differences design. Results revealed that differential parent-child conflict at age 11 years uniquely contributed to differential sibling EXT 3 years later but only in the most discordant twin pairs. In the full, unselected sample, this relationship was not significant. These results suggest that markedly different parent-child conflict has an environmentally mediated impact on child behavior through mid-adolescence, findings that yield insights into environmental influences on behavior. PMID- 17087562 TI - Examining genetic and environmental effects on reactive versus proactive aggression. AB - This study compared the contribution of genes and environment to teacher-rated reactive and proactive aggression in 6-year-old twin pairs (172 pairs: 55 monozygotic girls, 48 monozygotic boys, 33 dizygotic girls, 36 dizygotic boys). Genetic effects accounted for 39% of the variance of reactive aggression and for 41% of the variance of proactive aggression. The remainder of the variance was explained by unique environmental effects. Genetic as well as unique environmental effects were significantly correlated across reactive and proactive aggression (genetic correlation = .87, environmental correlation = .34), but this overlap was largely due to a common underlying form of aggression (i.e., teacher rated physical aggression). Once common etiological factors due to physical aggression were accounted for, reactive and proactive aggression shared no other genes and only a few environmental influences, although additional specific genetic and environmental effects were observed for both reactive and proactive aggression. These specific effects indicate that both reactive and proactive aggression may be influenced mostly by socialization experiences that are specific to each type of aggression and only to a very small degree by specific genes. PMID- 17087563 TI - A longitudinal examination of breadth and intensity of youth activity involvement and successful development. AB - Connections between youth activity involvement and indicators of successful development were examined in a longitudinal high school sample. Drawing on theories of expertise skill development (e.g., J. Cote, 1999); the selection, optimization, and compensation framework (P. B. Baltes, 1997); and theories of positive youth development (e.g., R. M. Lerner, J. B. Almerigi, C. Theokas, & J. Lerner, 2005), reciprocal associations between breadth and intensity of activity involvement and developmental success were explored. Time 1 breadth (but not intensity) and increases in breadth predicted higher levels of successful development at Time 2 (20 months later). Time 1 developmental success and improvements predicted greater Time 2 breadth and intensity. Implications for research and theory related to connections between youth activity involvement and successful development are discussed. PMID- 17087564 TI - Shared targets for aggression by early adolescent friends. AB - Similarity in early adolescent friends' general aggressiveness is well known, but questions remain regarding the degree to which friends aggress against the same victims. The authors examined this by administering the newly created Dyadic Aggression and Victimization Inventory to 417 sixth- through eighth-grade boys and girls (53%). Friends were found to share more targets for aggression than nonfriends, even after general levels of aggression were controlled (all ps < .05). Moreover, greater sharing of targets with friends relative to nonfriends was more pronounced among aggressive youths than nonaggressive youths, especially among aggressive youths' best friends relative to their other friends. Generally, these findings were similar across boys and girls as well as among older and younger youths. Implications of these findings and directions for future research are discussed. PMID- 17087565 TI - An attentional learning account of the shape bias: reply to Cimpian and Markman (2005) and Booth, Waxman, and Huang (2005). AB - Recently, Developmental Psychology published 2 articles on the shape bias; both rejected the authors' previous proposals about the role of attentional learning in the development of a shape bias in object name learning. A. Cimpian and E. Markman (2005) did so by arguing that the shape bias does not exist but is an experimental artifact. A. E. Booth, S. R. Waxman, and Y. T. Huang (2005), in contrast, concluded that the shape bias (and its contextual link to artifact categories) does exist but that the mechanisms that underlie it are conceptual knowledge and not attentional learning. In response, in this article the authors clarify the claims of the Attentional Learning Account (ALA) and interpretations of the data under question. The authors also seek to make explicit the deeper theoretical divide: cognition as sequestered from processes of perceiving and acting versus as embedded in, and inseparable from, those very processes. PMID- 17087566 TI - Deja Vu all over again: re-revisiting the conceptual status of early word learning: comment on Smith and Samuelson (2006). AB - The authors assert that L. B. Smith and L. Samuelson's (2006) most recent critique of A. E. Booth, S. R. Waxman, and Y. T. Huang's (2005) work missed its mark, deflecting attention from the important theoretical difference between the two sets of authors' positions and focusing instead on imagined differences and minor expositional complaints. The authors' goal in this response is twofold. First, they aim to redirect attention to the 1 clear difference between the 2 theoretical positions regarding word learning, a difference that is focused on the role of conceptual (in conjunction with perceptual) information in word learning. Second, they place L. B. Smith and L. Samuelson's (2006) current critique in the context of previous exchanges. PMID- 17087567 TI - Autonomy and relatedness with parents and romantic development in African American adolescents. AB - The influence of adolescents' autonomy and relatedness to parents on romantic relationships was examined longitudinally over 5 years in 76 middle-class African American late adolescents (mean age = 18.43 years). Relatedness to parents in early adolescence led to longer duration and more supportive romantic relationships in late adolescence, but longer duration was concurrently associated with more negative romantic relationships. Cluster analyses yielded 3 profiles of early adolescent autonomy and relatedness; early adolescents who were high in relatedness and low in autonomy to parents in early adolescence reported longer duration romantic relationships in late adolescence than did adolescents who were moderate or high in relatedness and high in autonomy. The implications for romantic development in African American middle-class youth are discussed. PMID- 17087568 TI - Get the picture? The effects of iconicity on toddlers' reenactment from picture books. AB - What do toddlers learn from everyday picture-book reading interactions? To date, there has been scant research exploring this question. In this study, the authors adapted a standard imitation procedure to examine 18- to 30-month-olds' ability to learn how to reenact a novel action sequence from a picture book. The results provide evidence that toddlers can imitate specific target actions on novel real world objects on the basis of a picture-book interaction. Children's imitative performance after the reading interaction varied both as a function of age and the level of iconicity of the pictures in the book. These findings are discussed in terms of children's emerging symbolic capacity and the flexibility of the cognitive representation. PMID- 17087569 TI - Distortions in the perceived lightness of faces: the role of race categories. AB - Although lightness perception is clearly influenced by contextual factors, it is not known whether knowledge about the reflectance of specific objects also affects their lightness. Recent research by O. H. MacLin and R. Malpass (2003) suggests that subjects label Black faces as darker than White faces, so in the current experiments, an adjustment methodology was used to test the degree to which expectations about the relative skin tone associated with faces of varying races affect the perceived lightness of those faces. White faces were consistently judged to be relatively lighter than Black faces, even for racially ambiguous faces that were disambiguated by labels. Accordingly, relatively abstract expectations about the relative reflectance of objects can affect their perceived lightness. PMID- 17087570 TI - A theory of dynamic occluded and illusory object perception. AB - Humans see whole objects from input fragmented in space and time, yet spatiotemporal object perception is poorly understood. The authors propose the theory of spatiotemporal relatability (STR), which describes the visual information and processes that allow visible fragments revealed at different times and places, due to motion and occlusion, to be assembled into unitary perceived objects. They present a formalization of STR that specifies spatial and temporal relations for object formation. Predictions from the theory regarding conditions that lead to unit formation were tested and confirmed in experiments with dynamic and static, occluded and illusory objects. Moreover, the results support the identity hypothesis of a common process for amodal and modal contour interpolation and provide new evidence regarding the relative efficiency of static and dynamic object formation. STR postulates a mental representation, the dynamic visual icon, that briefly maintains shapes and updates positions of occluded fragments to connect them with visible regions. The theory offers a unified account of interpolation processes for static, dynamic, occluded, and illusory objects. PMID- 17087571 TI - Visual sharpness contingency in recognition memory for orientation: mnemonic illusion suppressed by sensory signature. AB - A striking finding about human memory is that people's level of accuracy in remembering the orientation of heads on coins is often not simply at the chance level but significantly below it. However, S. W. Kelly, A. M. Burton, T. Kato, and S. Akamatsu (2001) reported that this is not so when two-alternative forced choice visual recognition is employed. The Kelly et al. result could not be replicated here with a copy of their stimuli. However, the result was successfully replicated with newly created stimuli. A series of experiments provided converging evidence that the mnemonic illusion is suppressed when recognition alternatives possess sharp visual detail. The role of a sensory signature in suppressing the mnemonic illusion and in modulating visual recognition performance in general is delineated. PMID- 17087573 TI - Retrieval-induced facilitation: initially nontested material can benefit from prior testing of related material. AB - Classroom exams can assess students' knowledge of only a subset of the material taught in a course. What are the implications of this approach for long-term retention? Three experiments (N = 210) examined how taking an initial test affects later memory for prose materials not initially tested. Experiment 1 shows that testing enhanced recall 24 hr later for the initially nontested material. This facilitation was not seen for participants given additional study opportunities without initial testing. Experiment 2 extends this facilitative effect to a within-subjects design. Experiment 3 demonstrates that this facilitation can be modulated by conscious strategies. These results have implications for educational practice and the theoretical developments of the testing effect, associative memory, and retrieval inhibition. PMID- 17087574 TI - Determinants of justification and self-control. AB - The authors propose that people use 2 routes in justifying self-gratification: 1st through hard work or excellence (entitlement) and the 2nd through the attainment of vices without depleting income. This framework was tested using real tasks and choices adopted from prior research on self-control. The results indicate that (a) higher effort and (bogus) excellence feedback increase preferences for vice rewards, but these effects are reversed or attenuated when the interchangeability of effort and income is implied; (b) willingness to pay in effort is greater for vices than virtues, but willingness to pay in income is higher for virtues; and (c) these effects are magnified among individuals with stronger (chronic or manipulated) guilt. The authors discuss the ability of the justification routes to explain the findings of prior self-control research. PMID- 17087575 TI - Hierarchical encoding of behavior: translating perception into action. AB - People encode goal-directed behaviors, such as assembling an object, by segmenting them into discrete actions, organized as goal-subgoal hierarchies. Does hierarchical encoding contribute to observational learning? Participants in 3 experiments segmented an object assembly task into coarse and fine units of action and later performed it themselves. Hierarchical encoding, measured by segmentation patterns, correlated with more accurate and more hierarchically structured performance of the later assembly task. Furthermore, hierarchical encoding increased when participants (a) segmented coarse units first, (b) explicitly looked for hierarchical structure, and (c) described actions while segmenting them. Improving hierarchical encoding always led to improvements in learning, as well as a surprising shift toward encoding and executing actions from the actor's spatial perspective instead of the participants' own. Hierarchical encoding facilitates observational learning by organizing perceived actions into a representation that can serve as an action plan. PMID- 17087576 TI - The association between psychological distance and construal level: evidence from an implicit association test. AB - According to construal level theory (N. Liberman, Y. Trope, & E. Stephan, in press; Y. Trope & N. Liberman, 2003), people use a more abstract, high construal level when judging, perceiving, and predicting more psychologically distal targets, and they judge more abstract targets as being more psychologically distal. The present research demonstrated that associations between more distance and higher level of construal also exist on a pure conceptual level. Eight experiments used the Implicit Association Test (IAT; A. G. Greenwald, D. E. McGhee, & J. L. K. Schwartz, 1998) to demonstrate an association between words related to construal level (low vs. high) and words related to four dimensions of distance (proximal vs. distal): temporal distance, spatial distance, social distance, and hypotheticality. In addition to demonstrating an association between level of construal and psychological distance, these findings also corroborate the assumption that all 4 dimensions of psychological distance are related to level of construal in a similar way and support the notion that they all are forms of psychological distance. PMID- 17087577 TI - Hierarchical control of cognitive processes: switching tasks in sequences. AB - Hierarchical control of cognitive processes was studied by examining the relationship between sequence- and task-level processing in the performance of explicit, memorized task sequences. In 4 experiments, switch costs in task switching performance were perturbed by sequence initiation times that varied with sequence complexity, preparation time, and type of sequence transition (repetition or switch). Hierarchical control was inferred from these sequence initiation time effects and the recurrent finding of no switch cost at the first serial position across sequences, the point at which sequence-level processes are likely active in maintaining or instantiating a hierarchical control structure in working memory. These findings resonate with past research on motor programs and serial memory and provide new insights into the concepts of task set and control. PMID- 17087578 TI - Seeing the forest when entry is unlikely: probability and the mental representation of events. AB - Conceptualizing probability as psychological distance, the authors draw on construal level theory (Y. Trope & N. Liberman, 2003) to propose that decreasing an event's probability leads individuals to represent the event by its central, abstract, general features (high-level construal) rather than by its peripheral, concrete, specific features (low-level construal). Results indicated that when reported probabilities of events were low rather than high, participants were more broad (Study 1) and inclusive (Study 2) in their categorization of objects, increased their preference for general rather than specific activity descriptions (Study 3), segmented ongoing behavior into fewer units (Study 4), were more successful at abstracting visual information (Study 5), and were less successful at identifying details missing within a coherent visual whole (Study 6). Further, after exposure to low-probability as opposed to high-probability phrases, participants increasingly preferred to identify actions in ends-related rather than means-related terms (Study 7). Implications for probability assessment and choice under uncertainty are discussed. PMID- 17087579 TI - Implicit task sets in task switching? AB - In 2 experiments, the authors compare stimulus-based versus task-rule-based task performance. Participants practiced 8 stimulus-response mappings either with or without knowledge about 2 underlying task sets. After practice, 2 transfer blocks with 8 new stimuli were presented. Results show that rule knowledge leads to significant switch and transfer costs, whereas without rule knowledge neither switch nor transfer costs occur. However, significant Task Type x Response Type interactions occurred in both conditions. In a second experiment including only the no rule condition, half of the stimulus-response mappings in the transfer blocks were incongruent to the underlying task rule. Slower response times for these incongruent stimuli as compared with congruent stimuli and the absence of switch costs suggest that participants acquired (presumably implicit) knowledge about 4 different stimulus-response categories. PMID- 17087580 TI - Proactive and retroactive effects of negative suggestion. AB - The negative effects of false information presented either prior to (proactive interference; PI) or following (retroactive interference; RI) true information was examined with word definitions (Experiment 1) and trivia facts (Experiment 2). Participants were explicitly aware of which information was true and false when shown, and true-false discrimination was evaluated via multiple-choice tests. Negative suggestion, defined as poorer performance on interference items than noninterference (control) items, consistently occurred when the wrong information followed the correct information (RI) but not when it preceded the correct information (PI). These effects did not change as a function of retention interval (immediate, 1 week, or 3 weeks) or number of incorrect alternatives (1 or 3). Implications of this outcome for experiencing incorrect information in both academic and nonacademic situations are considered. PMID- 17087581 TI - Linking associative and serial list memory: Pairs versus triples. AB - Paired associates and serial list memory are typically investigated separately. An "isolation principle" (J. B. Caplan, 2005) was proposed to explain behavior in both paradigms by using a single model, in which serial list and paired associates memory differ only in how isolated pairs of items are from interference from other studied items. In the present study, 2 experiments identify a critical dissociation between the 2 paradigms, challenging this unified account. Specifically, forward and backward probes were highly correlated for pairs and less so for short lists (triples). The authors asked whether the isolation principle could quantitatively accommodate this type of dissociation. A simulation confirmed that a single model incorporating the isolation principle can adequately explain this and other dissociations, supporting the common processes view. PMID- 17087582 TI - Collaboration in associative recognition memory: using recalled information to defend "new" judgments. AB - S. E. Clark, A. Hori, A. Putnam, and T. J. Martin (2000) showed that collaboration on a recognition memory task produced facilitation in recognition of targets but had inconsistent and sometimes negative effects regarding distractors. They accounted for these results within the framework of a dual process, recall-plus-familiarity model but showed only weak evidence to support it. The present results of 3 experiments present stronger evidence for Clark et al.'s dual-process view and also show why such evidence is difficult to obtain. PMID- 17087583 TI - Roles of egocentric and allocentric spatial representations in locomotion and reorientation. AB - Four experiments investigated the nature of spatial representations used in locomotion. Participants learned the layout of several objects and then pointed to the objects while blindfolded in 3 conditions: before turning (baseline), after turning to a new heading (updating), and after disorientation (disorientation). The internal consistency of pointing in the disorientation condition was relatively high and equivalent to that in the baseline and updating conditions, when the layout had salient intrinsic axes and the participants learned the locations of the objects on the periphery of the layout. The internal consistency of pointing was disrupted by disorientation when participants learned the locations of objects while standing amid them and the layout did not have salient intrinsic axes. It was also observed that many participants retrieved spatial relations after disorientation from the original learning heading. These results indicate that people form an allocentric representation of object-to object spatial relations when they learn the layout of a novel environment and use that representation to locate objects around them. Egocentric representations may be used to locate objects when allocentric representations are not of high fidelity. PMID- 17087584 TI - Perceptual representation as a mechanism of lexical ambiguity resolution: An investigation of span and processing time. AB - In 2 experiments, the authors investigated the ability of high- and low-span comprehenders to construe subtle shades of meaning through perceptual representation. High- and low-span comprehenders responded to pictures that either matched or mismatched a target object's shape as implied by the preceding sentence context. At 750 ms after hearing the sentence describing the target object, both high- and low-span comprehenders had activated a contextually appropriate perceptual representation of the target object. However, only high span comprehenders had perceptually represented the contextually appropriate meaning immediately upon hearing the sentence, whereas low-span comprehenders required more processing time before the perceptual representation was activated. The results are interpreted in a framework of co-occurring lexical representations and perceptual-motor representations. PMID- 17087585 TI - Similarity-based interference during language comprehension: Evidence from eye tracking during reading. AB - The nature of working memory operation during complex sentence comprehension was studied by means of eye-tracking methodology. Readers had difficulty when the syntax of a sentence required them to hold 2 similar noun phrases (NPs) in working memory before syntactically and semantically integrating either of the NPs with a verb. In sentence structures that placed these NPs at the same linear distances from one another but allowed integration with a verb for 1 of the NPs, the comprehension difficulty was not seen. These results are interpreted as indicating that similarity-based interference occurs online during the comprehension of complex sentences and that the degree of memory accessibility conventionally associated with different types of NPs does not have a strong effect on sentence processing. PMID- 17087586 TI - Readers' experiences of characters' goals and actions. AB - In this article, the authors examined readers' sensitivity to the match between characters' goals and characters' actions. In Experiment 1, readers integrated actions consistent with characters' goals more easily when there was a match between the extremeness of the actions and the urgency of the goals. In Experiments 2 and 3, characters' actions were consistent with either explicit or implicit goals. Participants showed different sensitivity to the mismatch between actions and urgent goals when they simply read the actions (Experiment 2) versus when they judged the likelihood of the actions (Experiment 3). Taken together, these results offer an account of how readers experience actions and goals when engaged in both local and global processing. PMID- 17087587 TI - Written spelling to dictation: Sound-to-spelling regularity affects both writing latencies and durations. AB - The authors examined the effect of sound-to-spelling regularity on written spelling latencies and writing durations in a dictation task in which participants had to write each target word 3 times in succession. The authors found that irregular words (i.e., those containing low-probability phoneme-to grapheme mappings) were slower both to initially produce and to execute in writing than were regular words. The regularity effect was found both when participants could and could not see their writing (Experiments 1 and 2) and was larger for low- than for high-frequency words (Experiment 3). These results suggest that central processing of the conflict generated by lexically specific and assembled spelling information for irregular words is not entirely resolved when the more peripheral processes controlling handwriting begin. PMID- 17087588 TI - Executive control in a modified antisaccade task: Effects of aging and bilingualism. AB - Two studies are reported that assess differences associated with aging and bilingualism in an executive control task. Previous work has suggested that bilinguals have an advantage over monolinguals in nonlinguistic tasks involving executive control; the major purpose of the present article is to ascertain which aspects of control are sensitive to the bilingual experience. Study 1 used an antisaccade task and found no effects of aging or bilingualism. Study 2 used the identical visual display but coupled to keypress responses. The results showed that bilinguals resolved various types of response conflict faster than monolinguals and that this bilingual advantage generally increased with age. A speculative interpretation of this pattern of results is offered in conclusion. PMID- 17087589 TI - Perspectives of probabilistic inferences: Reinforcement learning and an adaptive network compared. AB - The assumption that people possess a strategy repertoire for inferences has been raised repeatedly. The strategy selection learning theory specifies how people select strategies from this repertoire. The theory assumes that individuals select strategies proportional to their subjective expectations of how well the strategies solve particular problems; such expectations are assumed to be updated by reinforcement learning. The theory is compared with an adaptive network model that assumes people make inferences by integrating information according to a connectionist network. The network's weights are modified by error correction learning. The theories were tested against each other in 2 experimental studies. Study 1 showed that people substantially improved their inferences through feedback, which was appropriately predicted by the strategy selection learning theory. Study 2 examined a dynamic environment in which the strategies' performances changed. In this situation a quick adaptation to the new situation was not observed; rather, individuals got stuck on the strategy they had successfully applied previously. This "inertia effect" was most strongly predicted by the strategy selection learning theory. PMID- 17087590 TI - Go with the flow: How to master a nonlinear multiple-cue judgment task. AB - The authors examined the cognitive processes that participants use in linear and nonlinear multiple-cue judgment tasks, hypothesizing that people are unable to use explicit cue abstraction in a nonlinear task, instead turning to exemplar memory. Experiment 1 confirmed that people are unable to use cue abstraction in nonlinear tasks but failed to confirm the hypothesized, spontaneous shift to exemplar memory. Instead, the participants appeared to be trapped in persistent and futile attempts to abstract the cue-criterion relations. Only after being instructed to rely on exemplar memory in Experiment 2 did they master the nonlinear task. The results suggest that adaptive shifts of representation need not occur spontaneously and that analytical thought may sometimes harm performance in nonlinear tasks. PMID- 17087591 TI - Between ignorance and truth: Partition dependence and learning in judgment under uncertainty. AB - In 3 studies, participants viewed sequences of multiattribute objects (e.g., colored shapes) appearing with varying frequencies and judged the likelihood of the attributes of those objects. Judged probabilities reflected a compromise between (a) the frequency with which each attribute appeared and (b) the ignorance prior probability cued by the number of distinct values that the focal attribute could take on. Thus, judged probabilities were partition dependent, varying with the number of events into which the state space was subjectively divided. This bias was diminished among participants more confident in what they learned, was strong and insensitive to level of confidence when ignorance priors were especially salient, and required ignorance priors to be salient only when probabilities were elicited (not during encoding). PMID- 17087592 TI - Exemplar effects in the context of a categorization rule: Featural and holistic influences. AB - Brooks and colleagues (S. W. Allen & L. R. Brooks, 1991; G. Regehr & L. R. Brooks, 1993) have shown that the classification of transfer stimuli is influenced by their similarity to training stimuli, even when a perfect classification rule is available. It is argued that the original effect obtained by Brooks and colleagues might have resulted from two potential confounding variables. Once these confounds were controlled, the current authors did not replicate Brooks and colleagues' results in Experiment 1. Exemplar effects appeared in Experiment 2 when transfer stimuli were perceptually more similar to training stimuli than in Experiment 1. In Experiment 3, the authors obtained exemplar effects with separated stimuli, a finding that was not predicted by Brooks and colleagues' model. The authors suggest that a close perceptual match between training and transfer stimuli is necessary for the effect to occur, for both integrated and separated stimuli. The nature of this perceptual match, holistic or featural, is discussed. PMID- 17087593 TI - Producing biased diagnoses with unambiguous stimuli: The importance of feature instantiations. AB - In this article, the authors demonstrate a laboratory analogue of medical diagnostic biasing (V. R. LeBlanc, G. R. Norman, & L. R. Brooks, 2001) in 2 experiments and explore the basis of this effect. Before categorizing novel exemplars, participants first evaluated the likelihood that the item was a member of the category suggested on that trial: either the correct category or a plausible alternative category. This was sufficient to produce a substantial bias toward the suggested category despite the use of unambiguous stimuli, explicit rules, and unhurried conditions--each of which would be likely to limit diagnostic bias. The authors argue that the production of this effect requires distinguishing between particular feature instantiations and more abstract representations of those features as well as allowing people to adopt a particular decision strategy mediating the use of instantiated features: a feature-recognition heuristic. PMID- 17087594 TI - Concreteness and item-to-list context associations in the free recall of items differing in context variability. AB - Context variability can be defined as the number of preexperimental contexts in which a given concept appears. Following M. Steyvers and K. J. Malmberg's (2003) work, the authors have shown that concepts that are experienced in fewer preexperimental contexts generally are better remembered in episodic memory tasks than concepts that are experienced in a greater number of preexperimental contexts. The purpose of this article is to demonstrate that low context variability confers its memorial advantage because of stronger item-to-list context associations as compared with high context variability. Three experiments that use environmental context changes from study to test demonstrate that the low context variability advantage is eliminated when item-to-list context associations are not available because of environmental changes at test. In addition, the low context variability advantage is eliminated when inward processing at study prevents the formation of item-to-list context associations. PMID- 17087595 TI - Relation availability was not confounded with familiarity or plausibility in Gagne and Shoben (1997): Comment on Wisniewski and Murphy (2005). AB - C. L. Gagne and E. Shoben (1997) proposed that the conceptual system contains information about how concepts are used to modify other concepts and that this relational information influences the ease with which concepts combine. Recently, E. J. Wisniewski and G. L. Murphy suggested that C. L. Gagne and E. Shoben's measure of relation availability was confounded with familiarity and plausibility and that the participants could simply retrieve the stored meanings of the phrases because the phrases were not novel. In this article, the authors demonstrate that E. J. Wisniewski and G. L. Murphy's plausibility and familiarity judgments are dependent variables that (a) are themselves responsive to changes in relation availability, (b) modifier relation availability predicts response time even when the influence of phrase familiarity and plausibility is controlled, and (c) the materials consisted of mainly novel phrases. PMID- 17087597 TI - The effect of hyperthermia in vitro on vitality of rabbit preimplantation embryos. AB - The aim of our study was to test the influence of short exposure (6 h) of preimplantation rabbit embryos to elevated temperatures (41.5 degrees C or 42.5 degrees C) in vitro on their developmental capacity. Fertilized eggs recovered from female oviducts at the pronuclear stage (19 hpc) were cultured at standard temperature (37.5 degrees C) until the morula stage (72 hpc). Afterwards, the embryos were divided into two groups, cultured for 6 h either at hyperthermic (41.5 degrees C or 42.5 degrees C) or standard temperature (control 37.5 degrees C), post-incubated overnight (16-20 h) at 37.5 degrees C and then evaluated for developmental stages, apoptosis (TUNEL), proliferation (cell number), actin cytoskeleton and presence of heat-shock proteins Hsp70. It was observed that hyperthermia at 41.5 degrees C did not alter progression of embryos to higher preimplantation stages (expanded and hatching/hatched blastocysts), rate of apoptosis, total cell number of blastocysts and structure of actin filament compared to 37.5 degrees C. Western-blotting revealed the presence of heat stress induced 72 kDa fraction of Hsp70 proteins in granulosa cells (exposed to 41 degrees C) and embryos (exposed to 41.5 degrees C). Following the elevation of temperature to 42.5 degrees C embryo development was dramatically compromised. The embryos were arrested at the morula or early blastocyst stage, showed an increased rate of apoptosis and decreased total cell number compared to control. The structure of actin filaments in most of blastomeres was damaged and such blastomeres often contained apoptotic nuclei. In this group a presence of heat stress-induced fraction of Hsp70 proteins had not been confirmed. This is the first report demonstrating a threshold of thermotolerance of rabbit preimplantation embryos to hyperthermic exposure in vitro. A detrimental effect of higher temperature on the embryo is probably associated with the loss of their ability to produce Hsp70 de novo, which leads to cytoskeleton alterations and enhanced apoptosis. PMID- 17087598 TI - Differential effects of statins and alendronate on cholinesterases in serum and brain of rats. AB - Acetylcholinesterase (AChE) inhibitors represent standard treatment of Alzheimer's disease. Cholesterol plays an important role in Alzheimer's disease development. Because cholesterol synthesis may be inhibited by statins or bisphosphonates, we hypothesized that these drugs might possibly have an influence on cholinesterases. Moreover, we also evaluated if the cholesterol lowering agents that cross the blood-brain barrier (e.g. simvastatin) should be more effective than those which do not (e.g. atorvastatin). Four groups of rats were orally administered simvastatin, atorvastatin, alendronate or vehicle for seven days. Thereafter, blood samples were taken and the basal ganglia, septum, frontal cortex, and hippocampus were isolated from brains for measurement of acetylcholinesterase activity. In the blood, activities of neither acetyl- nor butyrylcholinesterase were influenced by any of the applied drugs. In the brain, no significant changes in AChE activity were observed after administration of atorvastatin. Both simvastatin and alendronate significantly suppressed the activity of AChE in the frontal cortex. In conclusion, our results confirmed the hypothesis that cholesterol-modifying drugs modulate AChE activity and it is more reasonable to use a blood-brain barrier penetrating drug. PMID- 17087599 TI - Renal adrenomedullin and high altitude diuresis. AB - Previous investigations revealed that most of the fluid regulating hormones showed no consistent relationship to the hypoxic diuretic response (HDR). In this study we examined if adrenomedullin (AM), a hypoxia-mediated diuretic/natriuretic peptide is connected to HDR. Thirty-three persons were examined at low altitude (LA), on the third exposure day at 3440 m (medium altitude, MA) and on the fourteenth day at 5050 m (high altitude, HA). Nocturnal diuresis rose from 460 ml [interquartile range 302 ml] at LA to 560 [660] ml at MA to 1015 [750] ml at HA (p<0.005). Sodium excretion was similar at LA and MA (41.8 [27.0] vs. 41.4 [28.4] mM) and increased to 80.2 [29.1] mM at HA (p<0.005). Urinary AM excretion was 7.9 [3.9] at LA, 7.5 [5.7] pM at MA, and increased to 10.5 [5.1] pM (p<0.05) at HA. Urinary AM excretion was correlated to diuresis (r=0.72, p<0.005) and sodium excretion (r=0.57, p<0.005). Plasma AM concentration rose from 16.4 [3.1] to 18.8 [4.9] pM/l at MA (p<0.005) and to 18.3 [4.3] pM/l at HA (p<0.005). Plasma AM concentration and urinary AM excretion were not correlated, neither were plasma AM concentration and diuresis or natriuresis. Our data suggest the involvement of increased renal AM production in the pathophysiology of high altitude fluid and sodium loss. PMID- 17087600 TI - Relation of cholesterol metabolism and non-cholesterol sterols to insulin resistance. AB - Using non-cholesterol sterols investigation several authors postulated a hypothesis that in the metabolic syndrome cholesterol endogenous synthesis is increased and its absorption decreased. Our study is the first attempt to evaluate the direct relation of cholesterol metabolism to the principal pathogenetic phenomenon of the metabolic syndrome--namely to insulin resistance. We have measured insulin sensitivity by two methods--Quicki (Quantitative Sensitivity Check Index) and intravenous insulin tolerance test (Kitt) and 3 indirect markers--fasting insulin level, fasting C-peptide level and SHBG (sex hormone binding globulin). The investigation was performed in three groups of subjects with a different prevalence of insulin resistance: 72 non-diabetics with ischemic heart disease, 117 young blood donors and 63 type 2 diabetics on diet therapy only. Analyzing altogether 60 relationships--between four sterols (lathosterol, squalene, sitosterol and campesterol) and five markers of insulin resistance in three groups of subjects--we have found only six significant relations between cholesterol synthesis and absorption and insulin resistance in all groups of patients. Our results indicate that there exists a significant relationship between insulin sensitivity and indices of either increased cholesterol synthesis or decreased cholesterol absorption. Insulin resistance explains only a part of both abnormalities mentioned above. PMID- 17087601 TI - Effect of PPAR-gamma agonist treatment on markers of endothelial dysfunction in patients with type 2 diabetes mellitus. AB - Thiazolidinediones are insulin-sensitizing drugs acting through peroxisome proliferator-activated receptor (PPAR)-gamma. The aim of our study was to evaluate the effect of 5-month treatment with PPAR-gamma agonist--rosiglitazone (4 mg/day), on the circulating markers of endothelial dysfunction and to evaluate the role of changes in endocrine function of adipose tissue in this process. Biochemical and metabolic parameters, circulating adiponectin, resistin, ICAM-1, VCAM-1, E-selectin, P-selectin, PAI-1, myeloperoxidase (MPO), and matrix metalloproteinase-9 (MMP-9) concentrations were assessed in 10 women with type 2 DM before and after rosiglitazone treatment and in a control group of healthy women. At baseline, diabetic group had significantly higher serum concentrations of glucose, glycated hemoglobin, V-CAM and PAI-1 compared to control group. Adiponectin levels tended to be lower in diabetic group, while resistin concentrations did not differ from control group. Rosiglitazone treatment improved diabetes compensation, significantly reduced VCAM-1, PAI-1 and E selectin concentrations and increased adiponectin levels, while it did not affect serum resistin concentrations. Adiponectin concentrations at baseline were inversely related to E-selectin and MPO levels, this correlation disappeared after rosiglitazone treatment. We conclude that 5-month rosiglitazone treatment significantly reduced several markers of endothelial dysfunction. This effect could be at least in part attributable to marked increase of circulating adiponectin levels. PMID- 17087602 TI - The role of tissue factor in thrombosis and hemostasis. AB - The tissue factor (TF) is one of the most important regulators of arterial thrombosis. Because arterial thrombosis is the pathophysiologic background of acute coronary syndrome, the possible impact of blocking the arterial thrombosis on its onset is a challenging problem. The investigations of TF brought a new concept of "cell-based coagulation model" which highlighted the question of blood borne TF as a source of TF in circulating blood. In this review we summarize essential information on the pathophysiology, molecular structure, expression and distribution of TF and we propose a novel concept of blood-borne TF, suggesting the possibilities of inhibition of the coagulation cascade with newly synthetized drugs. PMID- 17087603 TI - Physiological role of dendrotoxin-sensitive K+ channels in the rat cerebellar Purkinje neurons. AB - To understand the contribution of potassium (K+) channels, particularly alpha dendrotoxin (D-type)-sensitive K+ channels (Kv.1, Kv1.2 or Kv1.6 subunits), to the generation of neuronal spike output we must have detailed information of the functional role of these channels in the neuronal membrane. Conventional intracellular recording methods in current clamp mode were used to identify the role of alpha-dendrotoxin (alpha-DTX)-sensitive K+ channel currents in shaping the spike output and modulation of neuronal properties of cerebellar Purkinje neurons (PCs) in slices. Addition of alpha-DTX revealed that D-type K+ channels play an important role in the shaping of Purkinje neuronal firing behavior. Repetitive firing capability of PCs was increased following exposure to artificial cerebrospinal fluid (aCSF) containing alpha-DTX, so that in response to the injection of 0.6 nA depolarizing current pulse of 600 ms, the number of action potentials insignificantly increased from 15 in the presence of 4-AP to 29 action potentials per second after application of DTX following pretreatment with 4-AP. These results indicate that D-type K+ channels (Kv.1, Kv1.2 or Kv1.6 subunits) may contribute to the spike frequency adaptation in PCs. Our findings suggest that the activation of voltage-dependent K+ channels (D and A types) markedly affect the firing pattern of PCs. PMID- 17087604 TI - An experimental, non-uremic rabbit model of peritoneal dialysis. AB - Peritoneal dialysis (PD) is a well established method of depuration in uremic patients. Standard dialysis solutions currently in use are not biocompatible with the peritoneal membrane. Studying effects of dialysate on peritoneal membrane in humans is still a challenge. There is no consensus on the ideal experimental model so far. We, therefore, wanted to develop a new experimental non-uremic rabbit model of peritoneal dialysis, which would be practical, easy to conduct, not too costly, and convenient to investigate the long-term effect of dialysis fluids. The study was done on 17 healthy Chinchilla male and female rabbits, anesthetized with Thiopental in a dose of 0.5 mg/kg body mass. A catheter, specially made from Tro-soluset (Troge Medical GMBH, Hamburg, Germany) infusion system, was then surgically inserted and tunneled from animals' abdomen to their neck. The planned experimental procedure was 4 weeks of peritoneal dialysate instillation. The presented non-uremic rabbit model of peritoneal dialysis is relatively inexpensive, does not require sophisticated technology and was well tolerated by the animals. Complications such as peritonitis, dialysis fluid leakage, constipation and catheter obstruction were negligible. This model is reproducible and can be used to analyze the effects of different dialysis solutions on the rabbit peritoneal membrane. PMID- 17087605 TI - Effects of rate of decrease in power output in decrement-load exercise on oxygen uptake. AB - The purpose of this study was to examine how oxygen uptake (Vo2) in decrement load exercise (DLE) is affected by changing rate of decrease in power output. DLE was performed at three different rates of decrease in power output (10, 20 and 30 watts.min(-1): DLE10, DLE20 and DLE30, respectively) from power output corresponding to 90 % of peak Vo2. Vo2 exponentially increased and then decreased, and the rate of its decrease was reduced at low power output. The values of Vo2 in the three DLE tests were not different for the first 2 min despite the difference in power output. The relationship between Vo2 and power output below 50 watts was obtained as a slope to estimate excessive Vo2 (ex-Vo2) above 50 watts. The slopes were 10.0+/-0.9 for DLE10, 9.9+/-0.7 for DLE20 and 10.2+/-1.0 ml.min(-1).watt(-1) for DLE30. The difference between Vo2 estimated from the slope and measured Vo2 was defined as ex-Vo2. The peak value of ex-Vo2 for DLE10 (189+/-116 ml.min(-1)) was significantly greater than those for DLE20 and for DLE30 (93+/-97 and 88+/-34 ml.min(-1)). The difference between Vo2 in DLE and that in incremental-load exercise (ILE) below 50 watts (DeltaVo2) was greater in DLE30 and smallest in DLE10. There were significant differences in DeltaVo2 among the three DLE tests. The values of DeltaVo2 at 30 watts were 283+/-152 for DLE10, 413+/-136 for DLE20 and 483+/-187 ml.min(-1) for DLE30. Thus, a faster rate of decrease in power output resulted in no change of Vo2 at the onset of DLE, smaller ex-Vo2 and greater DeltaVo2. These results suggest that Vo2 is disposed in parallel in each motor unit released from power output or recruited in DLE. PMID- 17087606 TI - Does neonatal brain ischemia induce schizophrenia-like behavior in young adult rats? AB - Perinatal cerebral hypoxia represents a major cause of obstetric complications and the resulting transient oxygen deficiency might belong to early risk factors for schizophrenia. The aim of this study was to evaluate possible long-term behavioral changes induced by one hour of continuous bilateral common carotid artery occlusion in 12-day-old male rats. Post-ischemic behavioral disturbances were evaluated in social (play) behavior on postnatal day 22 (PND 22), open field test (PND 35 and 50) and prepulse inhibition of the acoustic startle reflex (PND 50). Transient ischemia in neonatal rats was not significantly altered in social dyadic interactions evaluated in pre-weaning pups, but resulted in enhanced locomotor activity in pubertal rats (PND 35) and impaired prepulse inhibition of the startle reflex in post-pubertal males (PND 50). These behavioral alterations suggest that perinatal hypoxic/ischemic insults may represent a risk factor for later manifestation of specific features relevant to schizophrenia in predisposed individuals. PMID- 17087607 TI - Association of C/T polymorphism in the LRP5 gene with circulating follicle stimulating hormone in Caucasian postmenopausal women. AB - The LRP5 gene is believed to be primarily associated with bone metabolism via Wnt signaling. The latter pathway, however, appears to control various other systems outside the skeleton. To find the relationships of the LRP5 gene to serum follicle stimulating hormone (FSH) and luteinizing hormone (LH) in the cohort of normal postmenopausal women, we identified the C/T (c.4037:A1330V) polymorphism in the LRP5 gene using a restriction analysis of the PCR product in a cohort of 165 untreated pre- and post-menopausal women. In a subset of 111 post-menopausal women we analyzed the association between the LRP5 genotype and serum levels of sex-hormones including FSH and LH. The distribution of CC, TC and TT genotypes of the C/T polymorphism in the whole group was 73.9 %, 23.6 % and 2.4 %, respectively, which is comparable with other Caucasian populations. As no TT homozygote was found in the group of post-menopausal women, serum sex-hormones were compared between CC and TC genotypes. Women with the CT allele combination had markedly higher serum FSH levels as compared to carriers of the CC genotype (p<0.004). No differences between these genotypes were found in serum LH levels as well as the circulating sex-steroids such as estradiol, testosterone, dehydroepiandrosterone and/or its sulphate, androstenedione and SHBG. To conclude, the LRP5 gene is associated with circulating FSH in normal post menopausal women in the present study. The mediating role of subtle undetectable variations in estrogen levels is discussed. We did not find any relationship between the LRP-5 genotype and serum LH levels. PMID- 17087608 TI - Relationship between antioxidant potential and oxidative damage to lipids, proteins and DNA in aged rats. AB - Oxidative stress has been implicated to play a major role in aging and age related diseases. In the present study, we investigated the effects of aging on the total antioxidant capacity, uric acid, lipid peroxidation, total sulfhydryl group content and damage to DNA in adult (6 months), old (15 months) and senescent (26 months) male Wistar rats. The antioxidant capacity, determined by phycoerythrin-based TRAP method (total peroxyl radical-trapping potential) was significantly decreased in the plasma and myocardium of old and senescent rats, whereas plasma level of uric acid was elevated in 26-month-old rats. Age-related decline in plasma and heart antioxidant capacity was accompanied by a significant loss in total sulfhydryl group content, increased lipid peroxidation and higher DNA damage in lymphocytes. Correlations between TRAP and oxidative damage to lipids, proteins and DNA suggest that the decline in antioxidant status may play an important role in age-related accumulation of cell damage caused by reactive oxygen species. PMID- 17087609 TI - Post-hoc analysis on the CD14 C(-260)T promoter polymorphism and coronary heart disease. AB - Functional C(-260)--> T polymorphism in the promoter of the CD14 gene has been reported to be associated with coronary heart disease (CHD). The functional role of the polymorphism, however, is still a matter of debate, since several studies have not proved its effect on clinical outcomes associated with atherosclerosis. Cardiovascular-related morbidity and mortality was assessed in a post-hoc approach four years after baseline characterization of patients (male/female n = 36/32) with angiographically proven coronary heart disease. CD14 C(-260)--> T promoter genotype was determined at baseline. Seventeen out of 20 CHD patients with non-lethal cardiovascular events carried at least one T-allele. CD14 T-260 allele carriers have a 3.59-fold (95 % confidence interval: 1.11-6.75) increased risk for non-lethal cardiovascular events (Kaplan-Meier plot: log rank test p = 0.029). All patients with lethal outcomes (n = 6) were also T-allele carriers. Multivariate logistic regression analysis among CHD patients including age, established risk factors and the C(-260)--> T polymorphism as covariates and non lethal events as a dependent variable confirmed the independent prospective effect of the T-allele on cardiovascular outcomes in this subset. Further evidence is provided for the role of CD14 C(-260)--> T promoter polymorphism as a genetic susceptibility marker of atherosclerosis in patients with an advanced clinical course of the disease. Due to the small sample size and post-hoc character of the study large-scale prospective studies that monitor patients with proven CHD are needed to confirm these findings. PMID- 17087611 TI - Influenza and pregnant women: hospitalization burden, United States, 1998-2002. AB - Women in later stages of pregnancy are at increased risk for serious influenza related morbidity; thus, universal influenza vaccination of pregnant women is recommended. However, vaccine uptake in the United States has been suboptimal. We previously described the burden of severe influenza-related morbidity during pregnancy in the United States by examining hospitalizations of pregnant women with respiratory illness during influenza season. Nondelivery hospitalizations with respiratory illness had significantly longer lengths of stay than those without respiratory illness. Hospitalization characteristics associated with greater likelihood of respiratory illness were the presence of a high-risk condition for which influenza vaccination is recommended, Medicaid/Medicare as primary expected payer, and hospitalization in a rural area. These findings may be explained by these women being at higher risk of influenza-related morbidity or reflect disparities in receipt of influenza immunization. Universal vaccination of pregnant women to decrease influenza-related morbidity should be encouraged. PMID- 17087612 TI - Toward optimal health: D. Casey Kerrigan, M.D., discusses the impact of footwear on the progression of osteoarthritis in women. Interview by Jodi R. Godfrey. PMID- 17087613 TI - Estrogen and androgen hormone therapy and well-being in surgically postmenopausal women. AB - BACKGROUND: Women undergoing surgical menopause experience an abrupt drop in gonadal hormones and are more likely to have symptoms that negatively impact well being, including hot flashes, sexual dysfunction, psychological problems, and testosterone deficiency. The purpose of this review was to examine the effects of hormone therapies on well-being among surgically menopausal women. METHODS: Studies were retrieved using both Cochrane and PubMed searches. A systematic literature review was performed to identify double-blind randomized controlled trials of the effects of menopausal hormone therapies on quality of life and well being among women who have undergone hysterectomy with bilateral oophorectomy. Two studies meeting these criteria were included for review. RESULTS: For each study reviewed, the following aspects were examined: type of hormonal therapies used, inclusion/exclusion criteria, overall changes, and changes in specific parameters of well-being. General well-being improved from baseline with certain types and doses of estrogen or estrogen plus testosterone therapy, with no serious adverse events. CONCLUSIONS: Estrogen with or without testosterone may improve general well-being in some groups of surgically menopausal women. Levels of serum estrogen achieved in these studies were within a normal range for premenopausal women. Adding testosterone to estrogen therapy may provide additional improvements in well-being in some women, but only at supraphysiological levels of total testosterone and physiological levels of free testosterone. It is recommended that the clinician discuss the potential benefits and risks with each woman and devise an individualized plan based on shared decision making. PMID- 17087614 TI - Physical activity patterns during pregnancy in a diverse population of women. AB - BACKGROUND: Participation in physical activity during pregnancy may reduce the risk of gestational diabetes mellitus and preeclampsia and help prevent excess maternal weight gain. However, studies describing patterns and correlates of activity during pregnancy are sparse. The purpose of this cross-sectional study was to describe total physical activity (household/caregiving, occupational, leisure, sports/exercise, and transportation) and correlates of total physical activity in a racially and economically diverse sample of 233 prenatal care patients. METHODS: Bilingual interviewers administered three 24-hour physical activity recalls among women in the first trimester (11%), second trimester (36%), and third trimester (53%) of pregnancy. RESULTS: Median total energy expenditure (MET-hours/day) was similar among women in the first and second trimesters (33.4 and 33.8 MET-hours/day, respectively) and was slightly, but not statistically significantly, lower among women in the third trimester (32.6 MET hours/day). Moderate intensity activity followed a similar pattern, being statistically significantly lower among women in the third trimester; vigorous intensity activity was low among women in each trimester of pregnancy. In terms of activity type, household/ caregiving activity was the largest contributor to both total and combined moderate and vigorous intensity energy expenditure among women in each trimester, constituting 24%-40% of total energy expenditure. Overall, total energy expenditure was highest in white non-Hispanic women and positively associated with increasing education and a history of previous live births (p < 0.01). CONCLUSIONS: Results from this study highlight the importance of including household/caregiving and occupational activities in addition to sports/exercise activities in the assessment of total energy expenditure during pregnancy. PMID- 17087615 TI - Health behaviors among American Indian/Alaska Native women, 1998-2000 BRFSS. AB - BACKGROUND AND OBJECTIVE: Minority populations, including American Indians and Alaska Natives (AI/AN), in the United States generally experience a disproportionate share of adverse health outcomes compared with whites. The prevalence of risk behaviors associated with these adverse health outcomes among AI/AN women is not well documented, especially for those who live outside areas serviced by Indian Health Service. We sought to describe the prevalence of selected health risk behaviors among AI/AN women, document the disparities between AI/AN women and all U.S. women, and demonstrate the efforts needed for AI/AN women to reach Healthy People 2010 goals. METHODS: Age-adjusted prevalence estimates for selected sociodemographic characteristics, current smoking, obesity, lack of leisure time physical activity, and binge drinking were calculated using Behavioral Risk Factor Surveillance System (BRFSS) data from 1998 to 2000, combined. Comparisons were made between prevalence estimates for AI/AN women and all women who participated in the BRFSS and Healthy People 2010 goals. RESULTS: The prevalences of current smoking (27.8%) and obesity (26.8%) were significantly higher among AI/AN women than among all U.S. women. AI/AN women did not meet Healthy People 2010 goals for current smoking, obesity, leisure time physical activity, or binge drinking. CONCLUSIONS: These data highlight both disparities in health risk behaviors between AI/AN women and all U.S. women and improvements needed for AI/AN women to meet Healthy People 2010 goals. This project demonstrates the overwhelming need for culturally appropriate and accessible prevention programs to address health risk behaviors associated with the leading causes of death among urbanized AI/AN women. PMID- 17087616 TI - The association between number of Pap smears performed and self-reported confidence in an internal medicine residency. AB - BACKGROUND: The American Board of Internal Medicine (ABIM) recommends internal medicine residents perform at least 3-5 Pap smears during training. We evaluated whether doing more than the required minimum Pap smears was associated with greater confidence and less desire for more Pap smear training. METHODS: We surveyed all 142 internal medicine residents at one university training program. Participants were asked how many Pap smears they had performed during residency and to rank their confidence and desire for more training in performing Pap smears. We compared confidence and desire for more training across three groups of residents reporting varying experience (10 Pap smears performed). RESULTS: Of 101 responding residents (71% eligible), 42 (42%), 19 (19%), and 36 (36%) reported performing 10 Pap smears, respectively. The number of Pap smears performed was significantly related to confidence and desire for more training in Pap smears (p<0.05). Of residents who reported 10 Pap smears were 14% and 17%, respectively. After adjusting for gender, primary care track, year of training, continuity clinic site, women's health rotation experience, and future plans in primary care, those reporting doing >10 Pap smears were significantly more likely to report confidence (OR 9.08, 95% CI 2.15-36.26) and less likely to want more training (OR 0.23; 0.06 0.93) than those reporting or=18.8 lb). Participants at 18 months were subdivided into women who had stayed on HT, 125 (28%); stopped HT after randomization, 145 (33%); and not on HT at baseline but stopped an average of 7 months prior to randomization, 173 (39%). Weight loss in the LC was similar for all three groups, but LDL lipoprotein response was better for women who stopped HT after randomization or were not on HT at baseline. CONCLUSIONS: The trial has been successful in increasing exercise and diet changes and reduction in weight and WC and variables related to metabolic syndrome. PMID- 17087621 TI - Hormones and quality of life after surgical menopause. PMID- 17087622 TI - Access to cervical cancer screening: training internists so skill limitations are not a barrier to care. PMID- 17087625 TI - Treatment of skeletally mature ovariectomized rhesus monkeys with PTH(1-84) for 16 months increases bone formation and density and improves trabecular architecture and biomechanical properties at the lumbar spine. AB - Histomorphometric studies of treatments for osteoporosis in humans are restricted to iliac crest biopsies. We studied the effects of PTH(1-84) treatment at the lumbar spine of skeletally mature ovariectomized rhesus monkeys. PTH increased bone turnover, rapidly normalized BMD, and increased vertebral compressive strength. PTH increased trabecular bone volume primarily by increasing trabecular number by markedly increasing intratrabecular tunneling. INTRODUCTION: Histomorphometric studies of the anabolic properties of PTH(1-84) (PTH) and related peptides in human bone are restricted to iliac crest biopsies. The ovariectomized (OVX) monkey is an accepted model of human postmenopausal bone loss and was used to study the effects of PTH treatment at clinically relevant skeletal sites. MATERIALS AND METHODS: Skeletally mature rhesus monkeys were OVX or sham-operated and, after a bone depletion period of 9 months, treated daily for 16 months with PTH (5, 10, or 25 microg/kg). Markers of bone formation (serum osteocalcin) and resorption (urine N-telopeptide [NTX]) and lumbar spine BMD were measured throughout the study. Trabecular architecture and vertebral biomechanical properties were quantified at 16 months. RESULTS: PTH treatment induced dose-dependent increases in bone turnover but did not increase serum calcium. Osteocalcin was significantly increased above OVX controls by 1 month. NTX was significantly elevated at 1 month with the highest dose, but not until 12 months with the 5 and 10 microg/kg doses. Lumbar spine BMD was 5% lower in OVX than in sham animals when treatment was started. All PTH doses increased BMD rapidly, with sham levels restored by 3-7 months with 10 and 25 microg/kg and by 16 months with 5 microg/kg. PTH treatment increased trabecular bone volume (BV/TV), primarily by increasing trabecular number, and dose-dependently increased bone formation rate (BFR) solely by increasing mineralizing surface. The largest effects on BV/TV and yield load occurred with the 10 microg/kg dose. The highest dose reduced trabecular thickness by markedly increasing intratrabecular tunneling. CONCLUSIONS: PTH treatment of OVX rhesus monkeys increased bone turnover and increased BV/TV, BMD, and strength at the lumbar spine. All PTH doses were safe, but the 10 microg/kg dose was generally optimal, possibly because the highest dose resulted in too marked a stimulation of bone remodeling. PMID- 17087626 TI - Deactivating germline mutations in LEMD3 cause osteopoikilosis and Buschke Ollendorff syndrome, but not sporadic melorheostosis. AB - Autosomal dominant OPK and BOS feature widespread foci of osteosclerotic trabeculae without or with skin lesions, respectively. Occasionally, a larger area of dense bone in OPK or BOS resembles MEL, a sporadic sclerosing disorder primarily involving cortical bone. Others, finding deactivating germline LEMD3 mutations in OPK or BOS, concluded such defects explain all three conditions. We found germline LEMD3 mutations in OPK and BOS but not in sporadic MEL. INTRODUCTION: In 2004, others discovered that heterozygous, loss-of-function, germline mutations in the LEMD3 gene (LEMD3 or MAN1) cause both osteopoikilosis (OPK) and Buschke-Ollendorff syndrome (BOS). OPK is an autosomal dominant, usually benign, skeletal dysplasia featuring multiple, small, especially metaphyseal, oval or round, dense trabecular foci distributed symmetrically throughout the skeleton. BOS combines OPK with connective tissue nevi comprised of collagen and elastin. In some OPK and BOS families, an individual may have relatively large, asymmetric areas of dense cortical bone interpreted as melorheostosis (MEL). MEL, however, classically refers to a sporadic, troublesome skeletal dysostosis featuring large, asymmetric, "flowing hyperostosis" of long bone cortices often with overlying, constricting soft tissue abnormalities. However, a heterozygous germline mutation in LEMD3 was offered to explain MEL. MATERIALS AND METHODS: We studied 11 unrelated individuals with sclerosing bone disorders where LEMD3 mutation was a potential etiology: familial OPK (1), familial BOS (2), previously reported familial OPK with MEL (1), sporadic MEL (3), sporadic MEL with mixed-sclerosing-bone dystrophy (1), and patients with other unusual sclerosing bone disorders (3). All coding exons and adjacent mRNA splice sites for LEMD3 were amplified by PCR and sequenced using genomic DNA from leukocytes. We did not study lesional tissue from bone or skin. RESULTS: In the OPK family, a heterozygous nonsense mutation (c.1433T>A, p.L478X) was discovered in exon 1. In the two BOS families, a heterozygous nonsense mutation (exon 1, c.1323C>A, p.Y441X) and a heterozygous frame-shift mutation (exon 1, c.332_333insTC) were identified. In the individual with MEL and familial OPK, a heterozygous nonsense mutation (c.1963C>T, p.R655X) was detected in exon 7. However, no LEMD3 mutation was found for any other patient, including all four with sporadic MEL. CONCLUSIONS: We confirm that OPK and BOS individuals, including those with MEL-like lesions, have heterozygous, deactivating, germline LEMD3 mutations. However, MEL remains of unknown etiology. PMID- 17087627 TI - Impaired angiogenesis, early callus formation, and late stage remodeling in fracture healing of osteopontin-deficient mice. AB - OPN is an ECM protein with diverse localization and functionality. The role of OPN during fracture healing was examined using wildtype and OPN(-/-) mice. Results showed that OPN plays an important role in regulation of angiogenesis, callus formation, and mechanical strength in early stages of healing and facilitates late stage bone remodeling and ECM organization. INTRODUCTION: Osteopontin (OPN) is an extracellular matrix (ECM) protein with diverse localization and functionality that has been reported to play a regulatory role in both angiogenesis and osteoclastic bone remodeling, two vital processes for normal bone healing. MATERIALS AND METHODS: Bone repair in wildtype and OPN(-/-) mice was studied using a femoral fracture model. microCT was used for quantitative angiographic measurements at 7 and 14 days and to assess callus size and mineralization at 7, 14, 28, and 56 days. Biomechanical testing was performed on intact bones and on fracture specimens at 14, 28, and 56 days. Histology and quantitative RT-PCR were used to evaluate cellular functions related to ECM formation and bone remodeling. RESULTS: OPN deficiency was validated in the OPN( /-) mice, which generally displayed normal levels of related ECM proteins. Intact OPN(-/-) bones displayed increased elastic modulus but decreased strength and ductility. Fracture neovascularization was reduced at 7 but not 14 days in OPN(-/ ) mice. OPN(-/-) mice exhibited smaller fracture calluses at 7 and 14 days, as well as lower maximum torque and work to failure. At 28 days, OPN(-/-) mice had normal callus size but a persistent reduction in maximum torque and work to failure. Osteoclast differentiation occurred normally, but mature osteoclasts displayed reduced functionality, decreasing late stage remodeling in OPN(-/-) mice. Thus, at 56 days, OPN(-/-) fractures possessed increased callus volume, increased mechanical stiffness, and altered collagen fiber organization. CONCLUSIONS: This study showed multiple, stage-dependent roles of OPN during fracture healing. We conclude that OPN deficiency alters the functionality of multiple cell types, resulting in delayed early vascularization, altered matrix organization and late remodeling, and reduced biomechanical properties. These findings contribute to an improved understanding of the role of OPN in vivo and provide new insight into mechanistic control of vascularization and bone regeneration during fracture repair. PMID- 17087629 TI - Critical issues in psychological autopsy studies. AB - This paper reviews research based on the psychological autopsy (PA) method applied to the study of suicide. It evidences the presence of a number of methodological problems. Shortcomings concern sampling biases in the selection of control subjects, confounding influences of extraneous variables, and reliability of the assessment instruments. The absence of homogeneity among studies in the procedure employed, as well as the lack of defined guidelines for performing this type of inquiry are emphasized. Questions needing empirical investigation in the future are pointed out. It is concluded that the validity and reliability of findings emerging from the use of this method of investigation would benefit from a standardization of its application. PMID- 17087630 TI - Standardizing the psychological autopsy: addressing the Daubert standard. AB - The psychological autopsy, as a research, clinical, and forensic tool, has gained widespread usage in suicidology over the last half century. In forensic settings, the lack of standardization and problems determining the procedure's validity and reliability pose significant issues for the procedure's admissibility under the Daubert standard of evidence. The Daubert standard requires that evidence must be founded on scientific knowledge and established five guidelines for judicial decisions regarding the admissibility of expert testimony. In this paper we examine expert opinion regarding the psychological autopsy and recommend a standardized protocol or template regarding areas and lines of inquiry for a psychological autopsy used in legal cases. PMID- 17087631 TI - The language of suicidology. AB - This 2005 Louis I. Dublin Award Address explores some of the basic difficulties and controversies inherent in the development and universal acceptance of a nomenclature for suicidology. Highlighted are some of the unresolved challenges with agreeing upon a mutually exclusive set of terms to describe suicidal thoughts, intentions, motivations, and self-destructive behaviors. PMID- 17087632 TI - On "intention" in the definition of suicide. AB - The need for a comprehensive nomenclature in suicidology is now well recognized. In this paper the focus is on the issue of intention, which is identified as an essential aspect of any definition of suicide and suicidal behavior primarily because of its distinction from accidental behavior. The distinction between the retrospective perspective of motives versus the prospective perspective of intentions is highlighted, and I argue that the latter is more closely related to suicidal behavior. Finally, while motives and intentions tend to be used together in research, there is a need for sound research to clarify the roles of intentions in order to better understand suicide and attempted suicide. PMID- 17087633 TI - Youth suicide risk factors and attitudes in New York and Vienna: a cross-cultural comparison. AB - The prevalence of suicide risk factors and attitudes about suicide and help seeking among New York and Viennese adolescents were compared in order to explore possible cross-cultural differences. Viennese adolescents exhibited higher rates of depressive symptomatology than their New York counterparts and had more first hand experience with suicidal peers. More attribution of suicide to mental illness was reported in Vienna; yet Viennese youth were less likely than New York adolescents to recognize the seriousness of suicide threats. Help-seeking patterns of Viennese adolescents were influenced by their setting a high value on confidentiality. These cross-cultural differences may reflect the limited exposure of Austrian youth to school-based suicide prevention programs. The findings highlight the need of taking the sociocultural context into consideration in the planning of youth suicide prevention strategies. PMID- 17087634 TI - Suicide and African American teenagers: risk factors and coping mechanisms. AB - In this study we investigated whether locus of control, hopelessness, and depression were primary risk factors for suicide ideation and attempts in African American youth, and whether congruency between locus of control and religious coping style reduced suicide risk. The sample consisted of 176 African American high school students (115 females; 61 males). Multiple and logistic regression were used to determine relationships among study variables. Depression was found to mediate the relationship between hopelessness and suicidal behaviors. External locus of control led to greater risk for hopelessness and depression, with a trend in the predicted direction for suicide attempts. Religious coping style alone was not always associated with reduction of risk of suicidal behaviors, yet some evidence suggests that congruency between locus of control and religious coping style reduces risk. Results provide additional support for suicide interventions to target depressive symptoms. Results also highlight the importance of examining the role of culturally salient variables such as fatalism and religious coping style when developing intervention programs for suicide. PMID- 17087635 TI - Suicidal behaviors and personality structure. AB - In this paper we suggest the potential clinical usefulness of two projective tests (Rorschach and Object Relation Technique) and of a clinical interview focused on the pathway to suicide, life events, and major life difficulties to better define subtypes of patients attempting suicide. Thirty-three hospitalized subjects who had attempted suicide in the previous 6 months were examined using an in-depth assessment of the pathway to suicide. The aims of this study were: (1) to compare, within a clinical sample of suicidal attempters, the psychopathological, personality, and psychosocial characteristics of subjects with mood disorders alone, personality disorders and dysfunctional personality alone, and mood disorders with personality disorders and/or dysfunctional personality; and (2) to analyze the relationship between the method chosen and the clinical characteristics of the subgroups. PMID- 17087636 TI - Hotel room suicide. AB - The objective of our research was to quantify the increased risk of suicide associated with registering in local hotels/motels. Medical examiner case files of suicide in King County, Washington, were reviewed for years 2002-2004. The incidence of suicide in local residents registering in local hotels/motels was 223/100,000 which is significantly greater than the incidence of suicide in the general population of King County (11.7/100,000 p < 0.0001). Hotel/motel guests from outside Washington had a significantly reduced incidence of suicide (3.9/100,000 p = 0.002). The study results suggest that there is an increased risk of suicide in local residents who register in local hotel rooms. PMID- 17087637 TI - Problem music and self-harming. AB - Academics and protest groups have claimed that "problem music" (hard rock, hip hop/rap, & punk) causes self-injurious thoughts/behaviors among fans. In this study we investigated whether the relationship is mediated by self-esteem, delinquency, and conservatism; and whether first exposure to problem music preceded self-injurious thoughts. A liking for problem music was associated with four of the five self-injurious measures, although these significant relationships were weakened (into nonsignificance in the case of two self injurious measures) when the mediating variables were included. Listening to problem music did not precede self-injurious thoughts. Problem music is associated with self-injurious thoughts and behaviors, but this relationship is mediated by other factors and the former does not seem to cause the latter. PMID- 17087638 TI - Sandro Morselli: schizophrenic solitude, suicide, and psychotherapy. PMID- 17087639 TI - Renal function--estimation of glomerular filtration rate. AB - Assessment and follow-up of renal dysfunction is important in the early detection and management of chronic kidney disease. The glomerular filtration rate (GFR) is the most accurate measurement of kidney disease and is reduced before the onset of clinical symptoms. Drawbacks to the measurement of GFR include the high cost and incompatibility with routine laboratory monitoring. Serum creatinine determination is a mainstay in the routine laboratory profile of renal function. The measurement of serum cystatin C has been proposed as a more sensitive marker for GFR. According to National Kidney Foundation-K/DOQ1 clinical guidelines for chronic kidney disease, serum markers should not be used alone to assess GFR. Based on prediction equations, clinical laboratories should report an estimate of GFR, in addition to reporting the serum value. In this article, information is presented on how best to estimate GFR using prediction equations for adults and for children. Using serum creatinine concentration with the Modification of Diet in Renal Disease (MDRD) study equation offers a suitable estimation of GFR in adults. The cystatin C prediction equation with the use of a prepubertal factor seems superior to creatinine-based prediction equations in children of <14 years. PMID- 17087640 TI - Research translation: a new frontier for clinical laboratories. AB - Translational research and translational medicine (referred to hereafter as translational research) are interchangeable terms that underline the pressing need to translate into practical benefits for those affected by disease the extensive investments divested by the private and public sectors in biomedical research. For people more directly involved in clinical practice (physicians, clinical laboratory professionals and patients), translational research responds to the need to accelerate the capture of benefits of research, closing the gap between what we know and what we practice. This basically means the transfer of diagnostic and therapeutic advances proven effective in large, well-conducted trials (and, therefore, evidence-based) to daily medical practice. Translational research should be regarded as a two-way road: bench to bedside, and bedside to bench. In particular, to make possible a more effective translation process, a new road map should be implemented through interaction and cooperation between basic researchers, clinicians, laboratory professionals and manufacturers. Some examples of recent developments in clinical laboratory testing, including markers of cardiovascular diseases, clinical proteomics and recombinant allergens, may explain the importance of careful evaluation of all variables that allow the introduction of such new insights into clinical practice to assure better clinical outcomes. The vital role of laboratory medicine in the delivery of safer and more effective healthcare requires more careful evaluation not only of the analytical characteristics, but also of any other variable that may affect the clinical usefulness and diagnostic performances of laboratory tests, thus allowing more accurate interpretation and utilization of laboratory information. PMID- 17087641 TI - Association of APOA5 c.553G>T polymorphism with type 2 diabetes mellitus in a Chinese population. AB - BACKGROUND: Several independent population studies have reported that c.553G>T polymorphism of the apolipoprotein A5 gene (APOA5) is associated with hypertriglyceridaemia. The aim of this study is to investigate the association between this genetic variation and the risk of type 2 diabetes mellitus. METHODS: In this study, APOA5 c.553G>T polymorphisms in 152 healthy individuals and 71 type 2 diabetes mellitus patients were detected by PCR-restriction fragment length polymorphism and agarose electrophoresis methods, and serum levels of lipids were also estimated by biochemical methods. RESULTS: The frequency of T alleles in the diabetes and control groups was 0.085 and 0.049, respectively. Compared with controls, there was no significant difference in the distribution of genotype and allele frequencies of c.553G>T polymorphic sites in diabetic patients (p=0.27 and p=0.15, respectively). However, the frequency of GT and TT genotypes and the T allele in the subgroup with hypertriglyceridaemia was significantly higher than that in the subgroup with normal triglyceridaemia in both the control group (p=0.034 and p=0.014, respectively) and the diabetes group (p=0.037 and p=0.007, respectively). In the diabetes and control groups, triglyceride levels in (GT+TT) genotype individuals were significantly higher than in GG genotype individuals (p=0.001 and p=0.003, respectively), and levels of low-density lipoprotein cholesterol were also significantly higher (p=0.044 and p=0.022, respectively). CONCLUSIONS: APOA5 c.553G>T polymorphism is not significantly associated with susceptibility to type 2 diabetes mellitus, but is associated with plasma triglyceride and low-density lipoprotein cholesterol levels. PMID- 17087642 TI - MTRR 66A>G polymorphism in relation to congenital heart defects. AB - BACKGROUND: Evidence is accumulating that periconceptional folic acid supplementation may prevent congenital heart defects (CHD). The methionine synthase reductase (MTRR) enzyme restores methionine synthase (MTR) enzyme activity and therefore plays an essential role in the folate- and vitamin B(12) dependent remethylation of homocysteine to methionine. We studied the influence of the MTRR 66A>G polymorphism on CHD risk. In addition, possible interaction between this variant and plasma methylmalonic acid (MMA) concentrations, as an indicator of intracellular vitamin B(12) status, was investigated. METHODS: Case control and case-parental studies were conducted to explore this association. In total, 169 CHD patients and 213 child controls, and 159 mothers with a CHD affected child and 245 female controls were included. RESULTS: The maternal MTRR 66AG and GG vs. AA genotypes revealed an odds ratio (OR) of 1.3 (95% CI 0.72 2.20) and 1.3 (0.71-2.37), respectively. Family-based transmission disequilibrium analysis did not reveal a significant association of the foetal 66G allele with the development of a heart defect in children (chi(2)=2.94, p=0.086). Maternal 66GG genotype in combination with high MMA concentration (above the 80th percentile) was associated with a three-fold (OR 3.3, 95% CI 0.86-12.50) increased risk for all types of CHD in offspring. CONCLUSIONS: These data indicate that maternal MTRR 66A>G polymorphism is not a risk factor for CHD. Maternal MTRR 66GG genotype with compromised vitamin B(12) status may possibly result in increased CHD risk. In addition to folate, vitamin B(12) supplementation may contribute to the prevention of CHD. PMID- 17087643 TI - Increased homocysteine in heart failure: a result of renal impairment? AB - BACKGROUND: Hyperhomocysteinemia may constitute a risk factor for patients with severe heart failure. This study examines the relationship between plasma homocysteine concentration and left ventricular ejection fraction with renal function in heart failure patients free of coronary artery disease. METHODS: Left ventricular ejection fraction was documented in 62 patients with advanced heart failure who had no proven significant coronary artery stenosis. Glomerular filtration rate was measured using the Cockroft-Gault equation. RESULTS: Elevated homocysteine levels (>or=15 micromol/L) were detected in 22 patients. Low glomerular filtration rate was observed in patients who had normal serum creatinine concentration. Homocysteine was strongly correlated with age, duration of disease, left ventricular ejection fraction, serum creatinine, and glomerular filtration rate. Statistically significant trends were observed across respective homocysteine quartiles. However, by multivariate regression, the strongest predictor of homocysteine was the glomerular filtration rate. CONCLUSIONS: Impaired renal function leads to a diminished clearance rate, which can be a prominent pathophysiological mechanism in the elevation of homocysteine concentration in heart failure. PMID- 17087644 TI - Urine flow cytometry and detection of glomerular hematuria. AB - BACKGROUND: The UF-100 is a flow cytometer designed for automated cellular urinalysis. In this study, the usefulness of the UF-100 in laboratory investigation into the origin of hematuria was evaluated. METHODS: Results from flow cytometric urinalysis were used to classify urinary red blood cells (RBCs) according to glomerular and non-glomerular origin and the classification was compared to the patient's clinical diagnosis as the gold standard. In parallel, microscopic sediment analysis was carried out. RESULTS: A total of 206 urine samples from 129 patients were analyzed (127 from patients with glomerular hematuria, 79 from patients with non-glomerular hematuria). Of these, 136 samples (92 patients) showed overt hematuria (>or=20 RBC/microL). Urine flow cytometry correctly classified 61% (sediment analysis 69%) of urine samples with overt hematuria. If inconclusive results are excluded, the UF-100 correctly diagnosed 85% (sediment analysis 98%) of urine samples with overt hematuria. The UF-100 and microscopic sediment analysis both showed sensitivity of 99% for the detection of glomerular hematuria. The specificity of the UF-100 for the detection of glomerular bleeding was lower (42%) than the specificity of microscopic sediment analysis (93%). CONCLUSIONS: Owing to its low specificity, the UF-100 showed limited capacity to discriminate glomerular from non-glomerular causes of hematuria in a population with a high incidence of renal disease. Therefore, extensive microscopic urinalysis remains necessary to assess the origin of hematuria. PMID- 17087645 TI - Chymotrypsin effects on the determination of sperm parameters and seminal biochemistry markers. AB - BACKGROUND: Few reports of the effects of treatment with chymotrypsin on the determination of sperm parameters and seminal biochemistry markers are documented. METHODS: Sperm parameters of 63 liquefied and 27 non-liquefied samples, untreated or treated with chymotrypsin, were evaluated using computer assisted semen analysis. In addition, biochemistry markers such as gamma glutamyltranspeptidase, alpha-glucosidase and fructose in 50 liquefied and 39 non liquefied samples, untreated or treated with chymotrypsin, were determined. RESULTS: Treatment with chymotrypsin had no effect on sperm concentration, motility, motility a and b, straightness, curvilinear velocity, straight line velocity, average path velocity and beat cross frequency in both liquefied and non-liquefied semen. However, linearity (p=0.025) decreased and the amplitude of the lateral head (p=0.029) increased significantly in non-liquefied semen after treatment with chymotrypsin. The levels of gamma-glutamyltranspeptidase, alpha glucosidase and fructose in seminal plasma were unaffected by chymotrypsin, regardless of liquefaction status. CONCLUSIONS: Chymotrypsin had no effects on the detection of sperm parameters and biochemistry markers, and could be used to treat non-liquefied samples before semen analysis in the andrology laboratory. PMID- 17087646 TI - Evaluation of cardiac involvement following major orthopedic surgery. AB - BACKGROUND: Cardiovascular morbidity is frequent after non-cardiac surgery and the early recognition of cardiac involvement is an essential tool for clinical risk stratification and management. The aim of this study was to investigate the behavior of traditional and emerging cardiac markers, including NT-prohormone brain natriuretic peptide (NT-proBNP) and ischemia-modified albumin (IMA), in the perioperative period in patients undergoing major uncomplicated orthopedic surgery. METHODS: A total of 37 patients undergoing major orthopedic surgery were longitudinally evaluated for NT-proBNP, IMA, cardiac troponin T (cTnT), creatine kinase isoenzyme MB and myoglobin 3 h before surgery and 4 and 72 h thereafter. RESULTS: NT-proBNP values were significantly increased at 72 h postoperative compared to both 3 h preoperative and 4 h postoperative (NT-proBNP: 20 vs. 4.5 pmol/L, p<0.001 and 20 vs. 5.9 pmol/L, p<0.001). IMA levels were significantly increased at 4 and 72 h postoperative vs. 3 h preoperative (132 vs. 113 kU/L, p=0.02 and 151 vs. 113 kU/L, p<0.001). In a stepwise regression model, the perioperative liquid amount and degree of modification in postoperative creatinine levels (delta-creatinine) were independently related to the NT-proBNP increase. CONCLUSIONS: The significant increase observed in NT-proBNP suggests that patients undergoing major uncomplicated orthopedic surgery may develop subclinical cardiac stress, presumably attributable to the considerable infusion of liquids. The clinical significance of this finding deserves further investigation, especially in patients at higher risk of heart failure. PMID- 17087647 TI - Increased sensitivity in detecting renal impairments by quantitative measurement of marker protein excretion compared to detection of pathological particles in urine sediment analysis. AB - BACKGROUND: Positive test strip results, pathological particles in urine and the presence of proteinuria are common findings in nephropathies. A comparison between these methods and renal biopsies became available with the introduction of quantitative measurement of marker proteins (albumin, transferrin, IgG, alpha(1)-microglobulin, retinol binding protein, alpha(2)-macroglobulin, Bence Jones proteins) and standardised urine sediment analysis by flow cytometry or microscopy. METHODS: A total of 400 urine samples were examined using marker protein patterns, test strips and quantitative sediment analyses. RESULTS: Results from standardised urine sediment analyses were compared with the excretion of renal marker proteins. Increased erythrocyte and leukocyte counts in urine were observed in only 29% and 39% of the samples for which pathological protein excretion was found. The sensitivity in detecting pathological particles in urine sediment, such as casts and/or dysmorphic erythrocytes, was only 19%. Renal biopsies from 65 patients who were classified as pathological were compared with proteinuria and sediment analyses. Increased excretion of marker proteins was found in all cases, whereas only 41% of the cellular urine sediments showed pathological results. CONCLUSIONS: Quantitative measurement of marker proteins from both the glomerular and tubular sides should be used upfront as screening parameters for the early detection of renal disorders. PMID- 17087648 TI - Clinical chemistry reference values for 75-year-old apparently healthy persons. AB - BACKGROUND: Clinical chemistry reference values for elderly persons are sparse and mostly intermixed with those for younger subjects. To understand the links between metabolism and aging, it is paramount to differentiate between "normal" physiological processes in apparently healthy elderly subjects and metabolic changes due to long-lasting diseases. The Vienna Transdanube Aging (VITA) study, which began in 2000 and is continuing, will allow us to do just that, because more than 600 male and female volunteers aged exactly 75 years (to exclude any influence of the "aging" factor in this cohort) are participating in this study. METHODS: Extensive clinical, neurological, biochemical, psychological, genetic, and radiological analyses, with a special emphasis on consumption of medication and abuse of drugs, were performed on each of the probands. The multitude of data and questionnaires obtained made possible an a posteriori approach to select individuals fulfilling criteria for a reference sample group of apparently healthy 75-year-old subjects for our study. Specific analytes were quantified on automated clinical analyzers, while manual methods were used for hormonal analytes. All clinical chemistry analytes were evaluated using in-depth statistical analyses with SPSS for Windows. RESULTS: In all, reference intervals for 45 analytes could be established. These include routine parameters for the assessment of organ functions, as well as hormone concentrations and hematological appraisals. Because all patients were reevaluated after exactly 30 months in the course of this study, we had the opportunity to reassess their health status at the age of 77.5 years. This was very useful for validation of the first round data set. Data of the second round evaluation corroborate the reference limits of the baseline analysis and further confirm our inclusion and exclusion criteria. CONCLUSIONS: In summary, we have established a reliable set of reference data for hormonal, hematological, and clinical chemistry analytes for elderly subjects. These values will be very useful for our future attempts to correlate disease states and aging processes with metabolic factors. PMID- 17087649 TI - Serum pro-hepcidin concentrations and their responses to oral iron supplementation in healthy subjects manifest considerable inter-individual variation. AB - Hepcidin participates in the regulation of iron homeostasis and its precursor pro hepcidin can be measured in serum. We evaluated pro-hepcidin serum concentrations in healthy subjects and the possible effects of iron supplementation on the results. The results suggest extensive physiological variation in serum pro hepcidin concentrations between healthy subjects with no symptoms or signs of anaemia, infections, inflammations, chronic disease or other interpretative factors. Before pro-hepcidin measurements can be used in clinical practise, further investigations are required to identify the physiological factors affecting normal serum pro-hepcidin variations in healthy subjects. The responses of serum pro-hepcidin to a 100-mg oral dose of iron also showed considerable inter-individual variation. In male subjects, no systematic changes in serum pro hepcidin concentrations were found and the increase in serum iron was fairly modest. In nine out of the ten female subjects who had rather low amounts of storage iron, iron supplementation was followed by an increase in both serum iron and serum pro-hepcidin concentrations. There were considerable inter-individual differences in the timing and magnitude of the response. We also evaluated the conceivable influences of sample storage and freeze-thaw cycles on the results of serum pro-hepcidin ELISA. We did not observe any changes in the results after serum samples were frozen and thawed up to four times and/or stored at room temperature for up to 6 h. PMID- 17087650 TI - Comparability of five analytical systems for the determination of triiodothyronine, thyroxine and thyroid-stimulating hormone. AB - BACKGROUND: The purpose of an external quality assessment (EQA) is to evaluate the analytical capability of clinical laboratories, identify differences among the laboratories and improve analytical quality. Our EQA results show that the rates of unsatisfactory performance for thyroid hormone tests were the highest in all of our EQA programs. Therefore, the main purpose of this study was to investigate unsatisfactory results by comparing the analytical values of five routinely used analytical systems. METHODS: The Kruskal-Wallis and two-sample Kolmogorov-Smirnov tests were used to identify analytical differences among and between analytical systems, respectively. RESULTS: The rates of significantly different results compared to the total number of analytical results were 81.1%, 64.5%, 93.3%, 50.0% and 56.7% for free triiodothyronine, total triiodothyronine, free thyroxine, total thyroxine and thyroid-stimulating hormone, respectively. CONCLUSIONS: Relatively large analytical differences between analytical systems were observed, especially when the analytical systems were used to measure free thyroid hormones. PMID- 17087651 TI - Automated analysis of pleural fluid total and differential leukocyte counts with the Sysmex XE-2100. AB - BACKGROUND: Determination of leukocyte (WBC) counts in pleural fluid is routinely performed by microscopic examination. In this study, we evaluated the performance of automated (differential) WBC counting in comparison with manual counting. METHODS: Pleural fluid samples (n=45) were obtained from patients undergoing diagnostic thoracocentesis. The manual total WBC count was determined after Samson staining in a Fuchs-Rosenthal hemocytometer; microscopic differential counts were performed on May-Grunwald Giemsa-stained cytospin slides. The Sysmex XE-2100 hematology analyzer was used for automated (differential) WBC counting. The functional detection limit was determined by serial dilution of continuous ambulatory peritoneal dialysis (CAPD) fluid and replicate measurements of each dilution. RESULTS: The automated WBC count (x10(6)/L) was highly correlated with that of the microscopic reference method (r(2)=0.95; WBC-analyzer=0.97 x WBC reference method+16; n=45). Good agreement was also observed for the absolute lymphocyte count (r(2)=0.92; WBC-analyzer=0.99 x WBC-reference method+32; n=36), neutrophil count (r(2)=0.94; WBC-analyzer=0.91 x WBC-reference method+6; n=35), and monocyte count (r(2)=0.73; WBC-analyzer=0.83 x WBC-reference method+6; n=38). The functional detection limit for WBCs was calculated at 50 x 10(6)/L (coefficient of variation 20%). CONCLUSIONS: With some limitations, total and differential WBC counts in pleural fluid can be reliably determined using the Sysmex XE-2100 instrument. PMID- 17087652 TI - Automation and validation of a fast method for the assessment of in vivo oxidative stress levels. AB - BACKGROUND: The d-ROMs test for the evaluation of serum hydroperoxides (HP) is simple, reliable, and cheap. Furthermore, it can easily be adapted to automated analyzers. Changing from the manual to an automated procedure allows the simultaneous processing of a large number of samples in a greatly reduced time, avoiding manual handling of samples and reagents and reducing variability sources. METHODS: This study was performed to adjust the manual procedure to a routine automated method in the clinical laboratory. We carried out the d-ROMs test in sera from 90 subjects of both sexes (34 men and 56 women) with age ranging from 20 to 80 years (mean 51+/-14 years). All subjects were free from acute or chronic inflammatory disease, immunological disease and history or evidence of malignancy. Subjects were not on vitamin and/or antioxidant therapies. RESULTS: The detection limit of the assay was 40 AU. Linearity was observed up to 475 AU. The recovery ranged between 97% and 105%. Within- and between-run imprecision was <5%. The mean HP value was 304+/-8 AU, with no significant difference between men (291+/-10 AU) and women (311+/-11 AU). A significant positive correlation was observed between age and HP in the whole population (r=0.4, p=0.0002). CONCLUSIONS: The automated test for the estimation of serum hydroperoxides represents a reliable and feasible procedure for increasing efficiency and reducing costs compared to the manual method, and is particularly suitable for evaluating oxidative stress in a variety of clinical conditions. PMID- 17087653 TI - Analytical validation of the new plasma calibrated Accu-Chek Test Strips (Roche Diagnostics). AB - BACKGROUND: The Accu-Chek Inform glucose monitor is a point-of-care system for testing blood glucose. New test strips, calibrated to deliver glucose plasma-like values, were launched on the market in May 2005. The aim of our study was to perform analytical validation of these new strips. METHODS: We compared the new plasma strips with whole blood strips; results for the plasma strips with plasma values obtained using a clinical analyzer and with whole blood values given by the glucose electrode of a blood gas analyzer; and the influence of the type of blood (capillary or venous) on the results obtained by the glucose monitor with the plasma calibrated strips. RESULTS: Plasma strips give on average 7% higher results than the previous whole blood strips. However, the results given by the plasma strips on capillary whole blood, even if well correlated, are not completely comparable with those given by an analyzer for venous plasma. Nevertheless, these plasma strips and the glucose electrode of a blood gas analyzer give comparable results. CONCLUSIONS: Accu-Chek Inform plasma strips are a good method for monitoring of blood glucose values in patients with diabetes. PMID- 17087654 TI - Use of insulin immunoassays in clinical studies involving rapid-acting insulin analogues: Bi-insulin IRMA preliminary assessment. AB - BACKGROUND: In clinical studies involving rapid-acting analogues (RAAs), insulin immunoreactivity is frequently measured, including endogenous, regular insulin (RI) and RAA immunoreactivities. Such a procedure implies equivalent cross reactivities of all insulins present in serum. Commercially available human insulin immunoassays have been widely used, but their limitations (including hemolysis and anti-insulin antibodies) were not fully investigated. The aims of our study were to compare cross-reactivities of RI and RAAs in buffer and in serum and to investigate insulin immunoassay pitfalls. METHODS: Cross reactivities were assessed using Bi-insulin IRMA (Schering Cis-Bio International) in phosphate-buffered saline (PBS)-1% bovine serum albumin (BSA) and in pools of sera spiked with RI and RAAs (lispro and aspart). To investigate the influence of hemolysis, a pool of sera spiked with RAA was mixed with a concentrated hemolysate (final hemoglobin concentration 10 g/L) and incubated for 3 h at room temperature. To determine interference by anti-insulin antibodies, insulin was removed using charcoal from 18 sera with anti-insulin antibodies and from 17 sera without detectable anti-insulin antibodies. These insulin-free samples were then spiked with RI and RAAs and the immunoreactivity was determined. RESULTS: Compared with buffer, cross-reactivity in serum for RI, lispro and aspart was lower (35%, 29% and 26% lower, respectively). Hemolysis degraded almost all RI and RAAs contained in the serum (>or=95%). Anti-insulin antibody interference was significant for RI and RAAs (p97% homogeneity in a single step in a biologically active state as judged by its efficient growth-promoting activity in Nb2 rat lymphoma cell proliferation assays. The expression and purification scheme, presented here, has a potential of scaling up to obtain pure and biologically active His-BbST relatively inexpensively for further studies and applications. PMID- 17087661 TI - Modulation of the proteolytic activity of matrix metalloproteinase-2 (gelatinase A) on fibrinogen. AB - The proteolytic processing of bovine fibrinogen by MMP-2 (gelatinase A), which brings about the formation of a product unable to form fibrin clots, has been studied at 37 degrees C. Catalytic parameters, although showing a somewhat lower catalytic efficiency with respect to thrombin and plasmin, indeed display values indicating a pathophysiological significance of this process. A parallel molecular modelling study predicts preferential binding of MMP-2 to the beta chain of fibrinogen through its haemopexin-like domain, which has been directly demonstrated by the inhibitory effect in the presence of the exogenous haemopexin like domain. However, the removal of this domain does not impair the interaction between MMP-2 and fibrinogen, but it dramatically alters the proteolytic mechanism, producing different fragmentation intermediates. The investigation at various pH values between 6.0 and 9.3 indicates a proton-linked behaviour, which is relevant for interpreting the influence on the process by environmental conditions occurring at the site of an injury. Furthermore, the action of MMP-2 on peroxynitrite-treated fibrinogen has been investigated, a situation possibly occurring under oxidative stress. The chemical alteration of fibrinogen, which has been shown to abolish its clotting activity, brings about only limited modifications of the catalytic parameters without altering the main enzymatic mechanism. PMID- 17087662 TI - Effects of glucagon-like peptide-1-(7-36)-amide on pancreatic islet and intestinal blood perfusion in Wistar rats and diabetic GK rats. AB - The aim of the present study was to evaluate the effects of GLP-1 [glucagon-like peptide-1-(7-36)-amide] on total pancreatic, islet and intestinal blood perfusion in spontaneously hyperglycaemic GK rats and normal Wistar rats using a microsphere technique. GK rats had hyperglycaemia and increased pancreatic and islet blood flow. Blood glucose concentrations were not affected when measured shortly (8 min) after GLP-1 administration in either GK or Wistar rats. GLP-1 had no effects on baseline pancreatic or islet blood flow in Wistar rats, but did prevent the blood flow increase normally seen following glucose administration to these animals. In GK rats, administration of GLP-1 decreased both pancreatic and islet blood flow. Glucose administration to the GK rats decreased pancreatic and islet blood flow. This decrease was not affected by pre-treatment with GLP-1. We conclude that administration of GLP-1 leads to a decrease in the augmented blood flow seen in islets of diabetic GK rats. The GLP-1-induced action on islet blood perfusion may modulate output of islet hormones and contribute to the antidiabetogenic effects of the drug in Type 2 diabetes (non-insulin-dependent diabetes). PMID- 17087664 TI - The national review of mental health nursing in Scotland: implications for the profession. PMID- 17087665 TI - Service users and carers' experiences of a psychosis service. AB - This paper presents the findings of a survey, which examined how people with psychosis and their carers had experienced local mental health services. The research was conducted prior to the establishment of an early intervention service and other service improvements in the region. The results suggest that the mental health service is improving overall for individuals with psychosis and their carers. The experiences of recent service users and carers seem more positive than those whose first contact with the services occurred during the 1970s and 1980s. Both service users and their carers report feeling largely supported by mental health professionals and voluntary support agencies, and feel included in the treatment approach. However, a number of important issues were highlighted. These include (i) the need for public education (which may help to reduce the delay which currently occurs between people seeking support and receiving treatment); (ii) improved access to alternative treatments, such as psychosocial therapies (in conjunction with pharmacological treatment); the provision of appropriate psychiatric facilities; and (iii) the inclusion of an early intervention treatment approach. The survey provides descriptive information from one NHS trust, which shows support for larger scale research findings and government recommendations. PMID- 17087666 TI - The national suicide prevention strategy for England: the reality of a national strategy for the nursing profession. AB - Suicide is recognized as a global phenomenon and many countries now have national suicide prevention strategies. International guidance on suicide prevention and accepted epidemiological and treatment-based research underpins healthcare policy relating to suicide reduction. There has been an established comprehensive strategy in England since 2002. However, the rate of suicide continues to be a concern and nurses hold a key role in the implementation of national, regional and local policy into practice. The aim of this paper is to consider the current implications of the national suicide prevention strategy in England for nursing. This discussion paper draws upon both empirical evidence-based literature, governmental guidance and policy-related documentation. The national suicide prevention strategy for England currently continues to have a multifaceted impact on the nursing profession. This ranges from clinical practice issues such as risk assessment through to broader public health responsibilities. If nurses and allied health professionals are to be effective in their role within suicide prevention, they will need to be supported in building awareness of the wider context of the national policy. In particular, this will mean working effectively and collaboratively with the voluntary sector, service users and other non medical agencies. PMID- 17087667 TI - Hope and schizophrenia: an integrative review. AB - The purpose of this article is to describe the current status of research on hope and schizophrenia. The CINAHL database was used to identify the articles that met the criteria. The searches were conducted using the terms 'hope', 'hope instillation (IOWA NIC)', 'hope (IOWA NOC)', 'schizophrenia' and their combinations. The findings were limited to research articles. In addition, Pub Medical database was used by searching the words 'hope' and 'schizophrenia' from the fields 'title' or 'abstract'. Four new articles were found. The data consist of 17 articles on hope and schizophrenia published in peer-reviewed journals, which were analysed using content analysis. Existing research has focused on people with schizophrenia (n = 8), significant others (n = 4), staff (n = 2), hope-engendering interventions (n = 2) and treatment evaluation related to hope (n = 2) in the care of people with schizophrenia. Different data collection methods have been used in these studies. The most common method was interview (n = 9), followed by questionnaires (n = 8) and observation (n = 1). Most studies used quantitative methods (n = 9). Hope is considered a positive factor in the life of a person living with schizophrenia, in significant others as well as in staff members. Existing research provides evidence of the following themes: factors associated with hope and factors contributing to hope in people with schizophrenia; hope from the perspective of significant others of people with schizophrenia; staff hopefulness and factors contributing to their hope, hope engendering interventions and treatment evaluation in regard to hope. Based on this review, research evidence of hope in the context of schizophrenia is quite scant and limited, even though the importance of hope in schizophrenia has been underlined in research reports and the literature. It is clear that hope is important to people with schizophrenia, their significant others and the healthcare personnel caring for them. It is therefore also important to study hope among these people. PMID- 17087668 TI - Moral perspectives on the prevention of suicide in mental health settings. AB - The response to suicidal clients is a recurrent and deep ethical problem in mental health practice. Rational suicide is rarely considered in relation to psychological suffering and is generally only discussed within the literature in connection with terminal illness. Focusing on a case example derived from a composite of patient experiences, this paper considers the premise that suicidal ideation may not be an irrational belief arising from mental disorder and analyses the ethical aspects of nursing care through the competing moral frameworks of the care-based and principle-based approaches to nursing ethics. We conclude that when the client is not capable of autonomous decision making, the two approaches lead to the same response. But when the client is capable of autonomous decision making, the two approaches lead to different responses. Specifically, from the care-based perspective, intervention to prevent suicide is easier to justify and helps formulate a nursing response, which promotes hope through the engagement. PMID- 17087669 TI - Group cognitive behavioural therapy for schizophrenia: a systematic review of the literature. AB - There are a growing number of studies which have evaluated group-based cognitive behavioural therapy (CBGT) for individuals with psychosis. This is at least partly driven by the need to maximize access to this evidence-based treatment which has still to be widely disseminated. In this paper we present a systematic review of trials which have evaluated the efficacy CBGT. Five controlled trials were identified, the results showed some improvements in specific areas of symptomatology such as social phobia and depression. However, methodological weaknesses of the studies compromised the confidence with which a cause and effect relationship can be established. CBGT may enable a greater number of individuals to be seen by a small group of trained therapists but whether this is an effective treatment remains to be established. In addition, whether therapists delivering cognitive behavioural therapy in a group format require a higher level of skill than those facilitating the intervention on an individual basis remains an empirical question. The finding that a short psycho-education group treatment reduced positive and negative symptoms is also worthy of further investigation given the difficulties reported in disseminating evidence-based psychosocial interventions for this client group. PMID- 17087670 TI - Illicit drug users in Northern Ireland: perceptions and experiences of health and social care professionals. AB - Over the 30 years of conflict, Northern Ireland escaped the worst excesses of illegal drug trafficking and usage. This was in large part due to the 'policing' of local community crime by paramilitaries. However, since the first 'cease-fire' in 1994 the province has witnessed a dramatic and unprecedented rise in the availability of illicit drugs. This increased availability reflects rising consumption of these drugs in the province. The aim of this study was to explore the perceptions that a sample of health and social care professionals' hold of illicit drug users in Northern Ireland. A survey methodology was used. Community settings across Northern Ireland were accessed. Focus groups and face-to-face interviews were used as data collection methods. Thirty-five health and social care professionals took part in this study. Thirty-two took part in six focus groups; three respondents underwent one-to-one interview. The participating professionals reported to have little or no education or training in the care and treatment of illicit drug users. Some stated that they would reject the offer of education and training in this area to prevent contact with illicit drug users. Many displayed strongly negative views of this client group, often expressing a preference not to care for or treat these people, preferring all their care to be given by specialists in illicit drug treatment. These findings indicate that most of the health and social care professionals who took part in this study appear unprepared and unwilling to meet the challenge of caring for illicit drug users. The findings will be of interest to service providers within and outside the UK. PMID- 17087671 TI - Sexual health education for people with mental health problems: what can we learn from the literature? AB - Research into sexual risk behaviour among people with 'severe' mental health problems suggested that they are likely to engage in high-risk sexual behaviour, for a number of reasons, putting them at risk of sexually transmitted diseases. The aim of this review is to describe approaches, content and outcomes of sexual health education programmes, developed and implemented for people with mental health problems. A literature review from 1980 to 2005 was carried out using the electronic databases CINAHL, PsycINFO, British Nursing Index, Pubmed and Medline, and the Cochrane library was also searched. The literature search was confined to papers written in English. The keywords 'sexuality', 'sexual health education', 'sexual health promotion', 'HIV', 'sexually transmitted disease' were combined with 'mental illness', 'chronic mental illness''severe mental illness''persistent mental illness''psychiatry', 'mental disorder', 'education interventions' and 'evaluation'. A vast amount of literature was recovered on sexual risk behaviour in people with severe mental health problems, and sexual dysfunction as a result of prescribed medication. As the focus of the review was on sexual health education, this literature was omitted. Although the literature on sexual health education for people experiencing mental health problems was sparse, 14 studies were located that either described or evaluated sexual health education programmes. Most sex education programmes focused on topics such as HIV and other sexually transmitted diseases, negotiating safe sex and skill development in condom use. Findings suggested that the people who attended benefited from sexual health education programmes, facilitated in a sensitive and supportive manner. Education tended to produce a reduction in sexual risk behaviour as opposed to complete cessation. Nevertheless, it is appropriate to consider integrating such education with service provision. The results of the review provide guidance to service providers and mental health nurses wishing to develop and evaluate sexual health education programmes for service users. Areas for future research are also identified. PMID- 17087672 TI - Potential of e-health in relation to depression: short survey of previous research. AB - E-health is developing at a high rate and represents an opportunity for the development and spreading of information and communication channels to interested parties. The aim of this study was to get an overview and comprehension of the e health field, with special focus on depression. A survey of initiatives and studies regarding e-health and depression was carried out. Relevant articles were found through searches on databases, search engines and reference lists. This paper shows that many different initiators with differing goals and motives are active within the e-health field. In the field of e-health and depression, the following areas show interesting results: studies mapping users' profile and habits, the quality of health-related information and the effectiveness of online therapies and supportive communities. Numerous initiators have launched different kinds of e-health initiatives. The potential of the Internet to be used constructively by health-care professionals and health-care consumers for health enhancing purposes still needs to be mapped, evaluated and developed. PMID- 17087673 TI - Quality of life for long-stay patients of psychiatric hospitals: a literature study. AB - The aim of this literature review is to provide an answer to the following questions: (1) What is known about the quality of life and quality of care for long-stay psychiatric patients? (2) What does literature say about possible contributions that health-care professionals may make to the quality of life for this target group? We conducted an open literature search across national and international databases, and added relevant literature by using cross-references. The quality of life and quality of care for long-stay psychiatric patients has been the subject of research on a limited scale only. Publications in this field provide a picture of patients with wishes and requirements that are generally basic and modest in nature. They also show that patients greatly value good relations with their care providers. This literature review shows that little is known about the quality of life of long-stay psychiatric patients and this review also shows that the relational aspects of care provide an eminent opportunity for care providers to contribute to the quality of life of their patients. Long-stay patients need care providers who are capable of developing and maintaining a caring relationship of the highest quality that will endure for many years. In their daily work, nurses have good opportunities to develop and maintain meaningful and reciprocal relationships with severely ill people, but this is a task that makes great demands on nurses in terms of both knowledge and interhuman skills. These skills should be given special attention both in nurse education and by supervisors. PMID- 17087674 TI - Problematizing special observation in psychiatry: Foucault, archaeology, genealogy, discourse and power/knowledge. AB - Special observation by mental health professionals is the recommended approach for those people deemed as at risk or risky. Recent research and academic writing have challenged the benefits of observing people/patients who are defined as 'at risk', and a more human engagement process is being recommended. Despite this assault, practice has not changed substantively, suggesting a need for a thorough exploration and questioning of the practices and process. The paper outlines three Foucaultian approaches to historical analysis. It applies aspects of Foucault's archaeology/genealogy, discourse and power/knowledge to explore the practices of special observation as a means of controlling risk, especially suicide risk. We identify the regulatory function of the 'gaze', professional codes and government policy in relation to restricting professional practices. We argue that observation can be related to moral therapy, wherein the person relinquishes madness for responsibility through a disciplinary process and, in governing risk, a 'professional industry' is created. The regulation of statements about people with mental health issues are exposed and related to what can be said and done by professionals. Finally, we look at productive power in relation to observation, and how it is intimately related to resistance. We conclude with 'soft' recommendations for practice discursively produced through the writing of the paper. PMID- 17087675 TI - Undergraduate psychiatric nursing education at the crossroads in Ireland. The generalist vs. specialist approach: towards a common foundation. AB - This paper provides a critical overview of undergraduate educational preparation for psychiatric nursing and proposes the use of a model, which may equip psychiatric nurses to adapt to current and future directions in psychiatric nursing practice. It contends that mental health nursing is a distinct scope of practice and as such requires specialty undergraduate education. A major difference between educational preparation for psychiatric nursing in the Republic of Ireland and the UK, and distinct from most other developed countries, is the existence of separate, specialized pre-registration programmes that, upon graduation, allow the nurse to register as a psychiatric nurse. In other countries such as, Australia, the integration of pre-registration nurse education into the university sector resulted in the wide-scale adoption of a generalist approach to nurse education. In light of the recent changes in pre-registration nursing education in Ireland, and the integration of nursing into higher-level education, this paper examines the generalist vs. specialist approach to nurse education. It contends that neither the generalist nor the specialist model best serve the nursing profession in preparing safe and competent practitioners. Rather, it argues that the use of a model which incorporates both generalist and specialist perspectives will help to redress the imbalance inherent in both of the existing approaches and promote a sense of unity in the profession without sacrificing the real strengths of specialization. Such a model will also facilitate psychiatric nurses to adapt to current and future directions in psychiatric nursing practice. PMID- 17087676 TI - The metabolic syndrome and schizophrenia: the latest evidence and nursing guidelines for management. AB - The introduction of second-generation antipsychotic drugs for the treatment of schizophrenia has provided significant benefits for clients experiencing this disorder. While they have been found effective in reducing psychotic symptoms, there is evidence that these drugs are also linked with a group of side effects commonly known as the metabolic syndrome. Mental health nurses are well positioned to prevent, detect and/or manage the development of this problematic constellation of symptoms. Guidelines for practice can be useful in prevention and management of the syndrome and enhance nursing care of clients who are taking second-generation antipsychotics. PMID- 17087677 TI - Living by themselves? Psychiatric nurses' views on supported housing for persons with severe and persistent mental illness. AB - The main principle directing the development of supported dwellings for persons with long-term mental illness is that to live in the community would improve their quality of life. The aim of this study was to describe psychiatric nurses' experiences of different types of supported dwelling for persons with long-term mental illness, and their views on what they consider to be important principles to provide for in order to facilitate their social integration into the community. Nine psychiatric nurses were interviewed. A qualitative content analysis revealed 'attempting to uphold the principle, respect for the patient's right to self-determination' as the main theme, which was linked to three sub themes: the nurses' view on their moral responsibility; the nurses' views on social norms that patients must follow in order to be accepted by their neighbours; and the nurses' views on supported dwelling of good quality. The nurses perceived that personal contact between the neighbour and the mentally ill person was one essential way to reduce fear of the mentally ill person. They viewed themselves as a link between the mentally ill person and other neighbours. Without the personal contact between the mentally ill person and the neighbours, there may be a risk that the integration will fail no matter how excellent the supported dwelling is framed. PMID- 17087678 TI - Views of mental illness and mental health care in Thailand: a report of an ethnographic study. AB - This paper reports some of the findings of an ethnographic study carried out in Thailand over a 2-year period. Interviews were conducted with three clinical nurses, three student nurses, 14 nurse educators, one psychiatrist, one Buddhist monk and two lay people (n = 24) about their views of mental health and mental health care in Thailand. Data (comprising field notes and interview transcripts) were analysed with the aid of Atlas.ti. Data were also collected through observation and conversation. This paper reports only the findings from the interviews. Findings emerged under the following headings: Causes of mental illness; Status of the mentally ill; Karma; Merit making; Kwan; Treatment and care; Reasons for becoming a mental health nurse. A range of causes, including the effects of ghosts and spirits, were identified under the first heading. The stigma of mental illness was noted under the second. Karma and merit making are Buddhist concepts and were discussed by many respondents as was the animist concept of kwan. Treatment and care seemed to include both 'modern' and 'traditional' approaches. These findings are discussed and some tentative 'rules' that appear to exist within the culture are mooted. The study is descriptive in nature and the findings cannot be generalized; however, it is hoped that they go some way to illuminate aspects of Thai culture as they relate to the mental health and mental health nursing fields. PMID- 17087679 TI - Closeness, chaos and crisis: the attractions of working in acute mental health care. AB - This paper makes a case for the attractiveness of acute mental health inpatient nursing (acute nursing) and argues that an altered perception of this work is essential if we are to provide the most acutely mentally ill and vulnerable people with a stable and expert nursing workforce. The discussion draws on an ethnographic study conducted in an inner-city psychiatric unit in England and the advantages of this method for understanding nursing work are described. Within our findings, we set out two overarching themes: the contextual realities of the contemporary acute ward and features of attraction that encourage nurses to work in the acute care setting. The former includes nurses' responsibility for the total ward environment and the latter the 'comfort of closeness' and 'surviving and thriving in chaos and crisis'. In conclusion, we argue that despite the unpopularity of the acute inpatient mental health environment, the highly sophisticated skills employed by acute nurses actually ensure the promotion of health for the majority of service users. PMID- 17087680 TI - Humanistic principles in relation to psychiatric nurse education: a review of the literature. AB - This paper addresses the topic of humanistic educative principles and examines it in the context of Irish psychiatric nurse education. Humanistic or existential philosophy influences the nursing curriculum profoundly, and yet a dichotomy continues to exist in relation to the epistemological basis informing nurse education. The dichotomy is manifested broadly in relation to the notion of individual choice and statutory responsibility as regulated professional governing bodies. Humanism is based on the notion that people are born 'blank slates' only to become who they are later. McKenna would describe the humanistic theory of learning as emphasizing feelings and experiences, leading to self awareness, personal growth and individual optimization. A review of the literature indicates there are very few empirical studies relating to the area of the humanistic principles as applied to psychiatric nurse education. Most of the literature that was initially located referred to humanistic existentialism in the field of psychology, which although provided interesting reading and relevant, to a point, did not apply specifically to psychiatric nursing education. The emphasis of this paper is psychiatric nursing education and these studies do not apply to the area under investigation. PMID- 17087681 TI - Re-examining the evaluation of interprofessional education for community mental health teams with a different lens: understanding presage, process and product factors. AB - This paper revisits the formative evaluation of a pilot project that offered in service interprofessional education (IPE), which is designed to enhance the collaborative practice, to two UK community mental health teams (CMHTs). While the IPE was well received and resulted in some improvements in team functioning, wider successes were elusive. Specifically, collaborative action plans were not implemented, and the pilot programme was ultimately not rolled out to other CMHTs. The purpose of this paper is to test the usefulness of the presage-process product (3P) framework for analysis as a means to untangle the complex web of factors that promoted and inhibited success in this initiative. The framework, which captures key features of the initiative as a dynamic system, proved effective, yielding new insights, making connections clearer and highlighting the critical importance of presage. We argue that use of the 3P model during the development of in-service IPE could ensure that planning oversights are minimized, thereby improving outcomes. PMID- 17087682 TI - The use of telemedicine in psychiatry. AB - This paper examines the potential effects of using video-conferencing within the field of mental health in the UK. In order to assess the usefulness of telepsychiatric services, an electronic search was conducted for articles published between August 1998 and July 2006 using the MEDLINE, EMBASE, PsychINFO and Telemedicine and Information Exchange (TIE) databases. The search was carried out using the following terms: telepsychiatry, videoconferencing and telepsychology. A total of 178 articles were identified and based on review of the abstracts 72 were identified as being specific to efficacy, cost effectiveness and satisfaction with psychiatric services delivered via videoconferencing. This paper concludes that the use of video conferencing can enhance psychiatric services within the UK especially for those patients who live in rural areas. Current advances in technology make this an increasingly more reliable and cost-effective method for assessing patients. The limitations of telemedicine are discussed and it is clear that this type of care is not suitable for all patients. Further research is required to assess the types of patients that telepsychiatry is most suitable for. PMID- 17087683 TI - Patient homicide causes nursing staff to leave. PMID- 17087684 TI - The forensic mental health nurse - a literature review. AB - Forensic mental health nursing has developed in the UK since the commissioning of Broadmoor Hospital in 1863. Little has been documented about the role until the early 1990s. There is debate over the usefulness of defining the role. It is suggested here that in order to meet the multifaceted, complex needs of the mentally disordered offender (MDO) patient group, the role of forensic mental health nurses (FMHNs) needs to be further developed, and this process is assisted by understanding its current status. This literature review examines the FMHN role in England and Wales. The literature pertinent to issues of task-orientated competence, knowledge and skills required, and desirable personal qualities that the role demands are examined. The latter of these attributes concerned with attitudes, values and morals is likely to be the most difficult to assess. Being able to isolate the composite elements of the role of FMHNs will enable the development of a competency framework, which will ensure quality assurance in contemporary health care for the thousands of FMHNs in practice and many more thousands of MDOs and other patients that the nurse has the potential to affect. PMID- 17087687 TI - Remembering Paul Beeson. PMID- 17087688 TI - Paul Beeson, MD: Today's Sir William Osler. PMID- 17087689 TI - The effects of in-home rehabilitation on task self-efficacy in mobility-impaired adults: A randomized clinical trial. AB - OBJECTIVES: To examine the effect on mobility self-efficacy of a multifactorial, individualized, occupational/physical therapy (OT/PT) intervention delivered via teletechnology or in-home visits. DESIGN: Randomized, clinical trial. SETTING: One Department of Veterans Affairs and one private rehabilitation hospital. PARTICIPANTS: Sixty-five community-dwelling adults with new mobility devices. Thirty-three were randomized to the control or usual care group (UCG), 32 to the intervention group (IG). INTERVENTION: Four, once-weekly, 1-hour OT/PT sessions targeting three mobility and three transfer tasks. A therapist delivered the intervention in the traditional home setting (trad group n = 16) or remotely via teletechnology (tele group n = 16). MEASUREMENTS: Ten-item Likert-scale measure of mobility self-efficacy. RESULTS: The IG had a statistically significantly greater increase in overall self-efficacy over the study period than the UCG (mean change: IG 8.8, 95% confidence interval (CI) = 3.8-13.7; UCG 1.2, 95% CI = 5.8-8.2). Descriptively, the IG exhibited positive changes in self-efficacy for all tasks and greater positive change than the UCG on all items with the exception of getting in and out of a chair. Comparisons of the two treatment delivery methods showed a medium standardized effect size (SES) in both the tele and trad groups, although it did not reach statistical significance for the tele group (SES: tele = 0.35, 95% CI = -2.5-0.95; trad = 0.54, 95% CI = 0.06-1.14). CONCLUSION: A multifactorial, individualized, home-based OT/PT intervention can improve self-efficacy in mobility-impaired adults. The trend toward increased self-efficacy irrespective of the mode of rehabilitation delivery suggests that telerehabilitation can be a viable alternative to or can augment traditional in home therapy. PMID- 17087690 TI - Alcohol intake and its relationship with bone mineral density, falls, and fracture risk in older men. AB - OBJECTIVES: To examine the association between alcohol intake and problem drinking history and bone mineral density (BMD), falls and fracture risk. DESIGN: Cross-sectional and prospective cohort study. SETTING: Six U.S. clinical centers. PARTICIPANTS: Five thousand nine hundred seventy-four men aged 65 and older. MEASUREMENTS: Alcohol intake and problem drinking histories were ascertained at baseline. Follow-up time was 1 year for falls and a mean of 3.65 years for fractures. RESULTS: Two thousand one hundred twenty-one participants (35.5%) reported limited alcohol intake (<12 drinks/y); 3,156 (52.8%) reported light intake (<14 drinks/wk), and 697 (11.7%) reported moderate to heavy intake (> or =14 drinks/wk) in the year before baseline. One thousand one men (16.8%) had ever had problem drinking. In multivariate models, as alcohol intake increased, so did hip and spine BMD (P for trend < .001). Greater alcohol intake was not associated with greater risk for nonspine or hip fractures. Men with light intake, but not moderate to heavy intake, had a lower risk of two or more incident falls (light intake: relative risk (RR) = 0.77, 95% confidence interval (CI) = 0.65-0.92; moderate to heavy intake: RR = 0.83, 95% CI = 0.63-1.10) than abstainers. Men with problem drinking had higher femoral neck (+1.3%) and spine BMD (+1.4%), and a higher risk of two or more falls (RR = 1.59; 95% CI = 1.30-1.94) than those without a history of problem drinking and similar total hip BMD and risk of fracture. CONCLUSION: In older men, recent alcohol intake is associated with higher BMD. Alcohol intake and fracture risk is unclear. Light alcohol intake may decrease the risk of falling, but a history of problem drinking increased fall risk. PMID- 17087691 TI - Can hip protector use cost-effectively prevent fractures in community-dwelling geriatric populations? AB - OBJECTIVES: To estimate the cost-effectiveness from a societal perspective of a hip protector (HP) program over the remaining lifetime of individuals initially living at home. DESIGN: A state-transition Markov model considering outcomes of HP use in cohorts stratified by age, sex, and functional and residential status. Costs, transition probabilities, HP adherence, and efficacy were derived from published sources. SETTING: Community and nursing homes in the United States. PARTICIPANTS: Hypothetical cohort of individuals aged 65 and older without a hip fracture and initially living at home. INTERVENTION: HP program. MEASUREMENTS: Fractures, life years, and dollars saved, quality-adjusted life-years (QALYs), and incremental cost-effectiveness ratios (ICER). RESULTS: HP use prevented fractures and increased life expectancy in all cohorts. HP use saved costs and improved QALYs in women initiating HP use at age 80 and in men at age 85. In women initiating HP use at age 75, the HP ICER was 19,000 dollars/QALY. In men initiating HP use at age 80, HP use saved costs but slightly decreased QALYs. In younger cohorts, HP use was neither cost saving nor QALY improving. In sensitivity analyses, if there was no QALY loss from wearing a HP, the ICER was less than 50,000 dollars/QALY for all age and sex cohorts. If HP cost was reduced 50%, HP use was cost saving for women initiating HP use at age 75. In probabilistic sensitivity analyses, the HP ICER was less than 50,000 dollars/QALY in 68% of simulations for women initiating HP use at age 75 and 61% of simulations for men initiating at age 85. CONCLUSION: HP use saved costs and QALYs for older age cohorts of both sexes. Additional research on the quality-of life effects and obstacles to wearing HP is warranted. PMID- 17087692 TI - Androgen treatment and muscle strength in elderly men: A meta-analysis. AB - OBJECTIVES: To review published, randomized trials examining the effect of androgen treatment on muscle strength in older men. DESIGN: Systematic review using meta-analysis procedures. SETTING: Computerized and manual searches. PARTICIPANTS: MEDLINE, EMBASE, CINAHL, and the Cochrane Register were searched for trials. Key words included testosterone, androgen, sarcopenia, muscle loss, aged, aging, elderly, older, geriatric, randomized controlled trials, and controlled clinical trials. Sixty-five nonoverlapping studies were found. Meta analysis methods were used to evaluate the 11 randomized, double-blind trials. INTERVENTION: Testosterone or dihydrotestosterone (DHT) replacement therapy in healthy men aged 65 and older. MEASUREMENTS: Tests of muscle strength. RESULTS: The studies included 38 statistical comparisons. The mean g-index (g(i)) adjusted for sample size was 0.53 (95% confidence interval (CI) = 0.21-0.86). Subanalyses revealed larger effects for measures of lower extremity muscle strength (g(i) = 0.63, 95% CI = 0.03-1.28) than for upper extremity muscle strength (g(i) = 0.47, 95% CI = 0.12-0.84). A larger mean g-index was found for injected (g(i) = 0.95, 95% CI = 0.33-1.58) than topical (g(i) = 0.26, 95% CI = 0.08-0.42) or oral (g(i) = -0.21, 95% CI = -1.40-1.02) administration of testosterone/DHT. Effect sizes were related to study characteristics such as subject attrition and design quality ratings. Sensitivity analyses revealed that the elimination of one study reduced the mean g-index from 0.53 to 0.23. CONCLUSION: The results suggest that testosterone/DHT therapy produced a moderate increase in muscle strength in men participating in 11 randomized trials. One study influenced the mean effect size. PMID- 17087693 TI - Identifying frailty in hospitalized older adults with significant coronary artery disease. AB - OBJECTIVES: To characterize physiological variation in hospitalized older adults with severe coronary artery disease (CAD) and evaluate the prevalence of frailty in this sample, to determine whether single-item performance measures are good indicators of multidimensional frailty, and to estimate the association between frailty and 6-month mortality. DESIGN: Observational cohort study. SETTING: Inpatient hospital cardiology ward. PARTICIPANTS: Three hundred nine consecutive inpatients aged 70 and older admitted to a cardiology service (n = 309; 70% male, 84% white) with minimum two-vessel CAD determined using cardiac catheterization. MEASUREMENTS: Two standard frailty phenotypes (Composite A and Composite B), usual gait speed, grip strength, chair stands, cardiology clinical variables, and 6-month mortality. RESULTS: Prevalence of frailty was 27% for Composite A versus 63% for Composite B. Utility of single-item measures for identifying frailty was greatest for gait speed (receiver operating characteristic curve c statistic = 0.89 for Composite A, 0.70 for Composite B) followed by chair-stands (c = 0.83, 0.66) and grip strength (c = 0.78, 0.57). After adjustment, composite scores and single-item measures were individually associated with higher mortality at 6 months. Slow gait speed (< or =0.65 m/s) and poor grip strength (< or =25 kg) were stronger predictors of 6-month mortality than either composite score (gait speed odds ratio (OR)=3.8, 95% confidence interval (CI) = 1.1-13.1; grip strength OR = 2.7, 95% CI = 0.7-10.0; Composite A OR = 1.9, 95% CI = 0.60-6.1; chair-stand OR = 1.5, 95% CI = 0.5-5.1; Composite B OR = 1.3, 95% CI = 0.3-5.2). CONCLUSION: Gait speed frailty was the strongest predictor of mortality in a population with CAD and may add to traditional risk assessments when predicting outcomes in this population. PMID- 17087694 TI - Functional impairment, race, and family expectations of death. AB - OBJECTIVES: To assess the effect of functional impairment on family expectations of death and to examine how this association varies by race. DESIGN: Cross sectional. SETTING: Community based. PARTICIPANTS: Two thousand two hundred thirty-seven family members of decedents from the Health and Retirement Survey (HRS), a national study of persons aged 50 and older. MEASUREMENTS: Families were interviewed within 2 years of the HRS participant's death. The primary outcome was whether death was expected. The primary predictors were the decedent's functional status (impairment in any activity of daily living (ADL; eating, dressing, transferring, toileting, or bathing) during the last 3 months of life and the decedent's race. RESULTS: Overall, 58% of families reported that their family member's death was expected. Expecting death was strongly associated with functional impairment; 71% of families of decedents with ADL disability expected death, compared with 24% of those without ADL disability (P < .01). Death was expected more often in families of white decedents (60%) than African Americans (49%) (P < .01), although the effect of ADL disability was similar in both groups. After adjustment for potentially confounding factors, there were still significant associations between expecting death and functional impairment (odds ratio (OR) = 3.58, 95% confidence interval (CI) 2.73-4.70), and families of African Americans expected death less often than families of white decedents (OR = 0.63, 95% CI = 0.46-0.86). CONCLUSION: Family members of older adults expected death only 58% of the time. Families of functionally impaired older people were more likely to expect death when it occurred than were families of older people who were not functionally impaired, and the expectation of death was lower for families of African Americans than for whites. PMID- 17087695 TI - Beyond substituted judgment: How surrogates navigate end-of-life decision-making. AB - OBJECTIVES: To characterize how surrogates plan to make medical decisions for others. DESIGN: Descriptive study using semistructured qualitative interviews. SETTING: Surrogates were interviewed by telephone from their homes. PARTICIPANTS: Fifty experienced surrogate decision-makers identified to make decisions for older, chronically ill veterans. MEASUREMENTS: Surrogates were asked to describe advance care planning conversations with loved ones and how they planned to make future medical decisions. Thematic content analysis was used to identify bases for decision-making. RESULTS: Surrogates described the motivators and the content of advance care planning conversations with loved ones. Surrogates described five bases for decision-making: (1) conversations (making decisions based on their knowledge of their loved ones' preferences), (2) relying on documents (referring to their loved ones' advance care directives), (3) shared experience (believing an "inner sense" would guide decisions because of shared lived experience with loved ones), (4) surrogates' own values and preferences about life, and (5) surrogates' network (enlisting the help of others). CONCLUSION: Although ethicists and clinicians expect surrogates to use substituted judgment or patients' best interests when making decisions, these data indicate that many surrogates rely on other factors such as their own best interests or mutual interests of themselves and the patient or intend to base substituted judgments on documents with which they have little familiarity. PMID- 17087696 TI - A multifaceted intervention to implement guidelines improved treatment of nursing home-acquired pneumonia in a state veterans home. AB - OBJECTIVES: To assess the feasibility of a multifaceted strategy to translate evidence-based guidelines for treating nursing home-acquired pneumonia (NHAP) into practice using a small intervention trial. DESIGN: Pre-posttest with untreated control group. SETTING: Two Colorado State Veterans Homes (SVHs) during two influenza seasons. PARTICIPANTS: Eighty-six residents with two or more signs of lower respiratory tract infection. INTERVENTION: Multifaceted, including a formative phase to modify the intervention, institutional-level change emphasizing immunization, and availability of appropriate antibiotics; interactive educational sessions for nurses; and academic detailing. MEASUREMENTS: Subjects' SVH medical records were reviewed for guideline compliance retrospectively for the influenza season before the intervention and prospectively during the intervention. Bivariate comparisons-of-care processes between the intervention and control facility before and after the intervention were made using the Fischer exact test. RESULTS: At the intervention facility, compliance with five of the guidelines improved: influenza vaccination, timely physician response to illness onset, x-ray for patients not being hospitalized, use of appropriate antibiotics, and timely antibiotic initiation for unstable patients. Chest x-ray and appropriate and timely antibiotics were significantly better at the intervention than at the control facility during the intervention year but not during the control year. CONCLUSION: Multifaceted, evidence-based, NHAP guideline implementation improved care processes in a SVH. Guideline implementation should be studied in a national sample of nursing homes to determine whether it improves quality of life and functional outcomes of this debilitating illness for long-term care residents. PMID- 17087697 TI - The relationship between sociodemographic factors and oral health-related quality of life in dentate and edentulous community-dwelling older adults. AB - OBJECTIVES: To quantify the associations between sociodemographic factors and oral health-related quality of life (OHRQoL) in dentate and edentulous community dwelling older adults. DESIGN: Cross-sectional study using a 54-item OHRQoL questionnaire. SETTING: Five counties in central Alabama: Jefferson and Tuscaloosa (urban), and Hale, Bibb, and Pickens (rural). PARTICIPANTS: Two hundred eighty-eight participants (Dental Study subjects) aged 65 and older were recruited from participants in the University of Alabama at Birmingham Study of Aging, a longitudinal study of mobility in older African Americans and non Hispanic whites. MEASUREMENTS: Dental Study subjects were queried on their OHRQoL and sociodemographic status and classified into two categories: dentate and edentulous. Bivariate analyses were used to quantify associations between sociodemographic variables and OHRQoL after excluding participants with severe depression. Variables included age, sex, race, marital status, veteran status, residence, income, education, and transportation difficulties. RESULTS: Dentate and edentulous subjects had similar OHRQoL across age, sex, marital status, veteran status, and residence. Analyses suggested a strong association between OHRQoL and race, education, income, and transportation difficulties in dentate subjects. Sociodemographic factors were less strongly associated with OHRQoL in edentulous participants. CONCLUSION: OHRQoL decrements were prevalent in dentate and edentulous subjects. Of dentate persons, African Americans and those with a 6th-grade education or less, with income less than 16,000 dollars/year, and with transportation difficulties were more likely to have decrements in OHRQoL. In edentulous persons, these associations were not statistically significant or were weaker. These findings suggest differential associations between sociodemographic factors and OHRQoL when stratified according to dentate status. PMID- 17087698 TI - Disparities between ambient, standard lighting and retinal acuities in community dwelling older people: Implications for disability. AB - OBJECTIVES: To examine three types of visual ability-ambient acuity, standard lighting acuity, and retinal acuity-and their relationship with self-reported disability. DESIGN: Cross-sectional prevalence survey. SETTING: New York, New York. PARTICIPANTS: Community-dwelling older people. MEASUREMENTS: Ambient acuity assessed using a near reading card; standard lighting acuity and retinal acuity assessed using the Retinal Acuity Meter. Difficulty in activities of daily living was assessed according to self-report. RESULTS: Mean logMAR acuities were 0.44 (20/56) for ambient acuity, 0.33 (20/44) for standard lighting acuity, and 0.19 (20/31) for retinal acuity (all pairwise differences, P < .001). Given the distribution of disability by ambient acuity in this sample, improving ambient acuity to the level of retinal acuity could potentially reduce self-care disability 22%. CONCLUSION: Better utilization of retinal acuity through optimal ophthalmologic care and improvement in lighting would likely reduce disability in older adults. PMID- 17087699 TI - Low alanine aminotransferase activity in older people is associated with greater long-term mortality. AB - OBJECTIVES: To find possible association between liver enzymes and mortality in older people. DESIGN: A prospective cohort study. SETTING: Jerusalem. PARTICIPANTS: A systematically selected representative sample of 455 70-year-old ambulatory individuals was prospectively followed for 12 years. MEASUREMENTS: An extensive social and medical profile was developed at age 70 using a detailed interview and physical and ancillary examination. Information on mortality was obtained annually. Differences in survival between subjects stratified according to liver enzyme levels were assessed using the Kaplan-Meier method. Multivariable survival analyses using a Cox proportional hazards model were performed to determine the association between liver enzyme levels at age 70 and mortality over 12 years. RESULTS: Median alanine aminotransferase (ALT) activity of the study population was 11.00 U/L for women and 13.00 U/L for men. Twelve-year survival rates for women with ALT below and above the median levels were similar (78%). For men, these rates were 54% and 65%, respectively (P < .001). Proportional hazards models demonstrated that this greater mortality risk was independent of numerous common risk factors for mortality (hazard ratio (HR) = 1.5, 95% confidence interval (CI) = 1.08-2.19). Adding an interaction between sex and low ALT to the model demonstrated a higher risk of mortality for men with low ALT levels (HR = 2.42, 95% CI = 1.15-5.08). No such risk was demonstrated for the other liver enzymes. CONCLUSION: ALT activity represents a strong and independent surrogate marker for mortality in community-dwelling elderly men. PMID- 17087700 TI - Use of herbal medicine and other dietary supplements in community-dwelling older people: Baseline data from the ginkgo evaluation of memory study. AB - OBJECTIVES: To analyze baseline data from the Ginkgo Evaluation of Memory (GEM) study, in which information was collected on the use of all dietary supplements. DESIGN: Cross-sectional regression analysis. SETTING: GEM study sites in California, Maryland, North Carolina, and Pennsylvania. PARTICIPANTS: The GEM study enrolled 3,072 ambulatory individuals aged 75 and older between September 2000 and June 2002. MEASUREMENTS: Self-reported use of dietary supplements and use identified through bottles brought to the clinic. RESULTS: Respectively, 59.4%, 66.6%, and 27.4% of the GEM study cohort used a multivitamin, at least one individual vitamin or mineral supplement, and some type of nonvitamin/nonmineral dietary supplement (NVNMDS). In logistic regression models, multivitamin use was associated with female sex, a higher income, a higher modified Mini-Mental State Examination score, difficulty with mobility, and asthma history; use of any other vitamin or mineral was associated with female sex, white race, nonsmoking, more years of schooling, difficulty walking, a history of osteoporosis, and reading health and senior magazines; and NVNMDS use was associated with residing in California, having difficulties with muscle strength, and reading health and senior magazines. CONCLUSION: There were substantial differences between individuals who used vitamins and minerals and those who used NVNMDS. These data require that trial investigators pay close attention to participant use of off protocol dietary supplements. In addition, these findings may help identify elderly individuals likely to combine NVNMDS and prescription drugs. PMID- 17087701 TI - Prevalence of perceived dysphonia in a geriatric population. AB - OBJECTIVES: To characterize geriatric dysphonia, including its prevalence, quality-of-life impairment, and association with overall health status. DESIGN: A validated survey-based study of geriatric dysphonia. SETTING: An independent living facility for geriatric individuals. PARTICIPANTS: The entire population of residents at the facility was offered the survey. The inclusion criterion was aged 65 and older. MEASUREMENTS: Two survey-based measures were used to characterize dysphonia: a direct question asking whether participants had problems with their voice and a voice-related quality-of-life (V-RQOL) measure. In addition, participants were administered the 12-item Medical Outcomes Study Short Form survey, U.S. version 2.0, a concise survey designed to evaluate overall health status. RESULTS: The prevalence of dysphonia was 20%. More than 50% of patients with voice problems incurred significant quality-of-life impairment resulting from their dysphonia as measured using V-RQOL scores. The mean total V-RQOL score+/-standard deviation was 89+/-20. Finally, general health measures did not reflect V-RQOL. CONCLUSION: There is a high prevalence of voice problems in older people, with a large proportion having significantly impaired quality of life related to their dysphonia. General health measures do not reflect V-RQOL, and many individuals may wrongly attribute dysphonia to age related change alone. Administration of validated instruments for assessing dysphonia is encouraged, because direct questions regarding voice difficulties may not be sensitive to the severity of vocal impairment. PMID- 17087702 TI - Clinical characteristics of obstructive sleep apnea in community-dwelling older adults. AB - OBJECTIVES: To examine whether traditional risk factors are common in older adults with obstructive sleep apnea (OSA). DESIGN: Cross-sectional study. SETTING: Atlanta, Georgia. PARTICIPANTS: A convenience sample of 94 community dwelling adults aged 62 to 91. MEASUREMENTS: Demographic, medical, and sleep related information obtained using questionnaires. Epworth Sleepiness Scale (ESS) and 72-hour voiding diary were used to determine daytime sleepiness and nocturia frequency, respectively. Overnight ambulatory sleep recording device was used to screen for OSA. RESULTS: Fifteen female and 15 male subjects had an apnea hypopnea index (AHI) of 15 or more per hour of sleep (moderate to severe OSA). Traditional risk factors such as snoring, body mass index, and neck circumference were not significantly associated with OSA. An AHI of 15 or more per hour was independently associated with not feeling well rested in the morning, higher ESS score, and greater frequency of nocturia. CONCLUSION: Traditional risk factors for OSA were not common presenting symptoms and signs in study subjects with an AHI of 15 or more per hour of sleep; this may contribute to underdiagnosis of OSA in this population. Subjects with an AHI of 15 or more per hour had more sleep related complaints and excessive daytime sleepiness. Although occult, this suggests that OSA may contribute to disease burden in this group of people. PMID- 17087703 TI - Cognitive correlates of financial abilities in mild cognitive impairment. AB - OBJECTIVES: To investigate the cognitive correlates of financial abilities in mild cognitive impairment (MCI). DESIGN: Controlled, matched-sample, cross sectional analysis regressing five cognitive composites on financial performance measures. SETTING: University medical and research centers. PARTICIPANTS: Forty three persons with MCI and 43 normal controls. MEASUREMENTS: The Financial Capacity Instrument (FCI) and a comprehensive neurocognitive battery. RESULTS: Patients with MCI performed significantly worse than controls on cognitive domains of executive function, memory, and language and on FCI domains of financial conceptual knowledge, bank statement management, and bill payment. Patients with MCI also needed significantly more time to complete a multistep financial task and were significantly more likely than controls to make errors on this task. Stepwise regression models revealed that, within the MCI group, attention and executive function were significant correlates of FCI performance. CONCLUSION: Although impaired memory is the cardinal deficit in MCI, the neurocognitive basis of lower functional performance in MCI appears to be emergent declines in abilities to selectively attend, self-monitor, and temporally integrate information. Compromised performance on cognitive measures of attention and executive function may constitute clinical markers of lower financial abilities and should be evaluated for its relationship to functional ability in general. These cognitive domains may be appropriate targets of future intervention studies aimed at preservation of functional independence in people with MCI. PMID- 17087704 TI - Association between reduced heart rate variability and cognitive impairment in older disabled women in the community: Women's Health and Aging Study I. AB - OBJECTIVES: To examine the independent association between heart rate variability (HRV), a marker of cardiac autonomic function, and cognitive impairment. DESIGN: Cross-sectional analysis of baseline data from Women's Health and Aging Study I. SETTING: Urban community in Baltimore, Maryland. PARTICIPANTS: A subset of 311 physically disabled, community-dwelling women aged 65 and older whose HRV data were obtained. MEASUREMENTS: Reduced HRV was defined as the lowest quartile of each of several HRV measures exploring time and frequency domains and compared with the remaining three quartiles. Cognitive impairment was defined as a Mini Mental State Examination score less than 24. Multiple logistic regression was used to model the independent relationship between reduced HRV and prevalent cognitive impairment. RESULTS: The age-, education-, and race-adjusted prevalence of cognitive impairment was higher in those with reduced HRV than in those with nonreduced HRV. After adjusting for relevant demographic and clinical characteristics, participants with reduced HRV were significantly more likely than those with nonreduced HRV to have cognitive impairment; these findings were consistent across different HRV indices. In particular, reduced high-frequency power, indicative of decreased parasympathetic activity, was associated with 6.7 times greater odds of cognitive impairment (95% confidence interval = 2.27-20.0). CONCLUSION: Cardiac autonomic dysfunction, particularly decreased parasympathetic activity, was independently associated with cognitive impairment in older disabled women in the community. This finding may improve understanding of the pathophysiological basis of cognitive impairment. The potential role of HRV as a cause or consequence of cognitive impairment needs to be elucidated in future studies. PMID- 17087705 TI - Socioeconomic status and survival in older patients with melanoma. AB - OBJECTIVES: To determine the association between socioeconomic status (SES) and survival in older patients with melanoma. DESIGN: Retrospective cohort study. SETTING: Surveillance, Epidemiology and End Results (SEER): a population-based cancer registry covering 14% of the U.S. population. PARTICIPANTS: Twenty-three thousand sixty-eight patients aged 65 and older with melanoma between 1988 and 1999. MEASUREMENTS: Outcome was melanoma-specific survival. Main independent variable was SES (measured as census tract median household income) taken from the SEER-Medicare linked data. RESULTS: Subjects residing in lower-income areas (< or =30,000 dollars/y) had lower 5-year survival rates (88.5% vs 91.1%, P < .001) than subjects residing in higher-income areas (>30,000 dollars/y). In Cox proportional hazard models, higher income was associated with lower risk of death from melanoma (hazard ratio = 0.88, 95% confidence interval = 0.79-0.98, P = .02) after adjusting for sociodemographics, stage at diagnosis, thickness, histology, anatomic site, and comorbidity index. There was an interaction effect between SES and ethnicity and survival from melanoma. For whites and nonwhites (all other ethnic groups), 5-year survival rates increased as income increased, although the effect was greater for nonwhites (77.6% to 90.1%, 1st to 5th quintiles, P = .01) than for whites (89.0% to 91.9%, 1st to 5th quintiles, P < .001). CONCLUSION: Older subjects covered by Medicare residing in lower-SES areas had poorer melanoma survival than those residing in higher-SES areas. Further research is needed to determine whether low SES is associated with late-stage disease biology and poorer early detection of melanoma. PMID- 17087706 TI - How should nations measure the quality of end-of-life care for older adults? Recommendations for an international minimum data set. AB - Nations face numerous challenges in providing high-quality end-of-life care for their citizens, and an aging population that has a prolonged trajectory of decline and substantial needs for care magnifies these challenges. As nations begin to address these challenges, it will be essential to determine whether their responses are effective, but current national data collection efforts are sporadic, and data are often collected only for patients who receive care through certain funding programs or provider organizations or in selected sites of care. Therefore, governments do not know whether their citizens are receiving adequate care, whether current programs are improving care, or how their outcomes compare with those of other countries. This article describes the importance of developing national minimum data sets that can be used to assess the quality of end-of-life care that older adults receive and recommends key variables that these data sets should include. PMID- 17087707 TI - Chronic low back pain in older adults: What physicians know, what they think they know, and what they should be taught. AB - Chronic low back pain (CLBP) is a common and debilitating problem in older adults. Little exists in the literature about primary care physicians' (PCPs') knowledge of and confidence in managing this problem. A self-administered survey was mailed to PCPs in western Pennsylvania to measure knowledge of the evaluation and treatment of common contributors to CLBP in older adults, confidence in diagnosing these contributors through physical examination, and the association between confidence levels and knowledge. The survey combined items with an ordinal scale on which PCPs ranked their confidence in detecting various contributors to CLBP (e.g., fibromyalgia) using physical examination and patient vignettes followed by multiple choice questions designed to assess knowledge. One hundred fifty-three of 634 surveys were returned (24.1%). Overall, the majority of PCPs did not feel "very confident" in their ability to diagnose any of the contributors of CLBP listed (most items <40%). PCPs felt most confident in detecting scoliosis and least confident detecting myofascial pain of the piriformis muscle. There was a wide range in the number of respondents answering all questions related to a particular topic correctly (3.9% for sacroiliac joint syndrome to 70.4% for hip osteoarthritis). There was no relationship between knowledge scores and confidence ratings (P > .05 for all comparisons). The results point to a need for more PCP education about CLBP in older adults. It also suggests that accurate needs assessment should not rely on physician confidence ratings alone. PMID- 17087708 TI - Reflections of medical students on visiting chronically ill older patients in the home. AB - The expanding number of Americans living with chronic illness necessitates educating future physicians about chronic illness care. Weill Cornell Medical College's Chronic Illness Care in the Home Setting Program (CIC-HSP), a mandatory part of the primary care clerkship, exposes medical students to persons with chronic illness via a half day of house calls with a geriatrics team. The investigators sought to qualitatively assess the effect of the CIC-HSP on medical students and recent medical graduates. Fifty-two prospective participants were approached, and 50 (96%) with varying training levels and time since completing the program were interviewed. Most respondents (63%) found that the home visits taught them important approaches to caring for the chronically ill, such as individualizing care to meet patients' individual needs and improving quality of life as a goal of care. Students remarked that the experience enhanced their empathy (18%) and sensitivity (20%) toward chronically ill patients and increased their appreciation for chronic illness care (35%). Many participants reported that patients were more empowered in the home (55%) and perceived greater rapport and warmth between the doctor and patient (57%) in the home (vs office) setting. The vast majority of recent medical graduates (84%) related that this educational exposure continued to positively influence their approach to patient care. A home visit experience with a geriatrics team can help foster medical students' understanding of the psychosocial and medical aspects of chronic illness, teach relevant approaches to patient care, and improve students' attitudes toward caring for the chronically ill. PMID- 17087709 TI - Functional disability in elderly Nigerians: Results from the Ibadan Study of Aging. AB - Functional disability is a common reason for loss of independence and need for informal care by elderly persons. There is little information on the profile of disability and the level of unmet need for care in elderly persons living in sub Saharan Africa. Using a multistage clustered sampling of households conducted in the Yoruba-speaking area of Nigeria (representing 22% of the national population), persons aged 65 and older (N = 2,152) were assessed for disability in activities of daily living and instrumental activities of daily living. Respondents were assessed for self-reports of physical health, for major depressive disorder, and for availability of informal care. The prevalence of any functional disability (defined as inability to independently perform any function) was 9.2% (standard error 0.6). In logistic regression analysis, high risks of disability were associated with female sex, older age, and urban dwelling. Risks were also high for persons with chronic pain, those with poor self-reported overall health, and those with evidence of undernutrition. Disabled persons had poorer quality of life and were more likely to suffer from major depressive disorder; 19.8% of disabled elderly persons lacked any informal care, and this unmet need for care increased the likelihood of having depression. The findings suggest a high burden of unmet need for care in a large section of disabled elderly persons in this African community undergoing demographic and social changes. Social factors relating to urbanization and poverty may be associated with the occurrence of disability and inability to access informal care. PMID- 17087710 TI - Formulary committees and Medicare Part D. PMID- 17087711 TI - High prevalence of diabetes mellitus in older people in a rural area in Laos. PMID- 17087712 TI - Carbocysteine therapy in older people with chronic obstructive pulmonary disease. PMID- 17087714 TI - Depressive symptoms in older people predict nursing home admission. PMID- 17087715 TI - Self-neglect in older adults is a geriatric syndrome. PMID- 17087717 TI - Osteoporosis in patients with Parkinson's disease. PMID- 17087718 TI - An unusual complication of anticoagulation therapy in an elderly patient: Pituitary apoplexy with remission of acromegaly. PMID- 17087719 TI - Vascular parkinsonism in an elderly woman with mixed cryoglobulinemia associated with hepatitis C infection. PMID- 17087720 TI - Whole-diet approach and risk of chronic disease: Limits and advantages. PMID- 17087722 TI - Delirium was likely due to multiple factors, rather than gatifloxacin induced. PMID- 17087723 TI - cAMP increases mitochondrial cholesterol transport through the induction of arachidonic acid release inside this organelle in Leydig cells. AB - We have investigated the direct effect of arachidonic acid on cholesterol transport in intact cells or isolated mitochondria from steroidogenic cells and the effect of cyclic-AMP on the specific release of this fatty acid inside the mitochondria. We show for the first time that cyclic-AMP can regulate the release of arachidonic acid in a specialized compartment of MA-10 Leydig cells, e.g. the mitochondria, and that the fatty acid induces cholesterol transport through a mechanism different from the classical pathway. Arachidonic acid and arachidonoyl CoA can stimulate cholesterol transport in isolated mitochondria from nonstimulated cells. The effect of arachidonoyl-CoA is inhibited by the reduction in the expression or in the activity of a mitochondrial thioesterase that uses arachidonoyl-CoA as a substrate to release arachidonic acid. cAMP-induced arachidonic acid accumulation into the mitochondria is also reduced when the mitochondrial thioesterase activity or expression is blocked. This new feature in the regulation of cholesterol transport by arachidonic acid and the release of arachidonic acid in specialized compartment of the cells could offer novel means for understanding the regulation of steroid synthesis but also would be important in other situations such as neuropathological disorders or oncology disorders, where cholesterol transport plays an important role. PMID- 17087724 TI - Human cationic trypsinogen is sulfated on Tyr154. AB - The crystal structure of human pancreatic cationic trypsin showed the chemical modification of Tyr154, which was originally described as phosphorylation [Gaboriaud C, Serre L, Guy-Crotte O, Forest E & Fontecilla-Camps JC (1996) J Mol Biol259, 995-1010]. Here we report that Tyr154 is sulfated, not phosphorylated. Cationic and anionic trypsinogens were purified from human pancreatic juice and subjected to alkaline hydrolysis. Modified tyrosine amino acids were separated on a Dowex cation-exchange column and analyzed by thin layer chromatography. Both human cationic and anionic trypsinogens contained tyrosine sulfate, but no tyrosine phosphate, whereas bovine trypsinogen contained neither. Furthermore, incorporation of [(35)S]SO(4) into human cationic trypsinogen transiently expressed by human embryonic kidney 239T cells was demonstrated. Mutation of Tyr154 to Phe abolished radioactive sulfate incorporation, confirming that Tyr154 is the site of sulfation in cationic trypsinogen. Sulfated pancreatic cationic trypsinogen exhibited faster autoactivation than a nonsulfated recombinant form, suggesting that tyrosine sulfation of trypsinogens might enhance intestinal digestive zymogen activation in humans. Finally, sequence alignment revealed that the sulfation motif is only conserved in primate trypsinogens, suggesting that typsinogen sulfation is absent in other vertebrates. PMID- 17087725 TI - Characterization of a second proliferating cell nuclear antigen (PCNA2) from Drosophila melanogaster. AB - The eukaryotic DNA polymerase processivity factor, proliferating cell nuclear antigen, is an essential component in the DNA replication and repair machinery. In Drosophila melanogaster, we cloned a second PCNA cDNA that differs from that encoded by the gene mus209 (for convenience called DmPCNA1 in this article). The second PCNA cDNA (DmPCNA2) encoded a 255 amino acid protein with 51.7% identity to DmPCNA1, and was ubiquitously expressed during Drosophila development. DmPCNA2 was localized in nuclei as a homotrimeric complex and associated with Drosophila DNA polymerase delta and epsilonin vivo. Treatment of cells with methyl methanesulfonate or hydrogen peroxide increased the amount of both DmPCNA2 and DmPCNA1 associating with chromatin, whereas exposure to UV light increased the level of association of only DmPCNA1. Our observations suggest that DmPCNA2 may function as an independent sliding clamp of DmPCNA1 when DNA repair occurs. PMID- 17087726 TI - Leishmania infantum LeIF protein is an ATP-dependent RNA helicase and an eIF4A like factor that inhibits translation in yeast. AB - LeIF, a Leishmania protein similar to the eukaryotic initiation factor eIF4A, which is a prototype of the DEAD box protein family, was originally described as a Th1-type natural adjuvant and as an antigen that induces an IL12-mediated Th1 response in the peripheral blood mononuclear cells of leishmaniasis patients. This study aims to characterize this protein by comparative biochemical and genetic analysis with eIF4A in order to assess its potential as a target for drug development. We show that a His-tagged, recombinant, LeIF protein of Leishmania infantum, which was purified from Escherichia coli, is both an RNA-dependent ATPase and an ATP-dependent RNA helicase in vitro, as described previously for other members of the DEAD box helicase protein family. In vivo experiments show that the LeIF gene cannot complement the deletion of the essential TIF1 and TIF2 genes in the yeast Saccharomyces cerevisiae that encode eIF4A. In contrast, expression of LeIF inhibits yeast growth when endogenous eIF4A is expressed off only one of its two encoding genes. Furthermore, in vitro binding assays show that LeIF interacts with yeast eIF4G. These results show an unproductive interaction of LeIF with translation initiation factors in yeast. Furthermore, the 25 amino terminal residues were shown to enhance the ability of LeIF to interfere with the translation machinery in yeast. PMID- 17087727 TI - Identification of bikunin as an endogenous inhibitor of dynorphin convertase in human cerebrospinal fluid. AB - Dynorphin-converting enzymes constitute a group of peptidases capable of converting dynorphins to enkephalins. Through the action of these enzymes, the dynorphin-related peptides bind to delta-opioid instead of kappa-opioid receptors, leading to a change in the biological function of the neuropeptides. In this article, we describe the identification of the protein bikunin as an endogenous, competitive inhibitor of a dynorphin-converting enzyme in human cerebrospinal fluid. This protein is present together with its target enzyme in the same body fluids. The K(M) value of the convertase was found to be 9 microm, and the K(i) value of the inhibitor was 1.7 nm. The finding indicates that bikunin may play a significant role as a regulatory mechanism of neuropeptides, where one bioactive peptide is converted to a shorter sequence, which in turn, can affect the action of its longer form. PMID- 17087728 TI - Effect of von Willebrand disease type 2B and type 2M mutations on the susceptibility of von Willebrand factor to ADAMTS-13. AB - BACKGROUND: von Willebrand disease (VWD) type 2 is associated with mutations in von Willebrand factor (VWF) that affect its secretion, multimeric pattern, affinity for platelet receptors and clearance of the protein. While increased proteolysis by a disintegrin-like and metalloprotease with thrombospondin type 1 motifs-13 (ADAMTS-13) has been clearly established for VWF type 2A, only little is known about VWF types 2B and 2M in this regard. OBJECTIVES: Sensitivity of wild-type (WT) and mutated recombinant (r) VWF to proteolysis by ADAMTS-13 was investigated to better understand the role of this process in the pathophysiology of VWD. METHODS: We used human rADAMTS-13-WT to digest 11 full-length recombinant forms of VWF carrying molecular abnormalities identified in patients with VWD type 2A (E1638K and P1648S), type 2B (InsM1303, R1306W, R1308P and V1314F) and type 2M (G1324A, E1359K, K1362T, R1374H and I1425F). RESULTS: Using low ionic strength conditions, all mutations induced increased proteolysis of rVWF by rADAMTS-13 as compared with rVWF-WT. The susceptibility of mutants decreased in the following order: type 2A > type 2B > type 2M > rVWF-WT. At physiological salt concentration (150 mm NaCl) the sensitivity of all rVWF to rADAMTS-13 was significantly decreased. However, type 2A and type 2B mutants still exhibited a significantly higher susceptibility to rADAMTS-13 than rVWF-WT, whereas type 2M mutants normalized. CONCLUSIONS: Type 2M mutants and rVWF-WT exhibit a similar sensitivity to rADAMTS-13-mediated proteolysis, in agreement with the normal multimeric pattern in vivo. In VWD type 2B, the spontaneous binding to platelets and excessive degradation by ADAMTS-13 of VWF high-molecular-weight multimers may account for their clearance from plasma. PMID- 17087729 TI - The decrease of fibrinogen is an early predictor of the severity of postpartum hemorrhage. AB - BACKGROUND: Postpartum hemorrhage (PPH) is a major source of maternal morbidity. OBJECTIVES: This study's objective was to determine whether changes in hemostasis markers during the course of PPH are predictive of its severity. PATIENTS AND METHODS: We enrolled 128 women with PPH requiring uterotonic prostaglandin E2 (sulprostone) infusion. Two groups were defined (severe and non-severe PPH) according to the outcome during the first 24 hours. According to our criteria, 50 of the 128 women had severe PPH. Serial coagulation tests were performed at enrollment (H0), and 1, 2, 4 and 24 hours thereafter. RESULTS: At H0, and through H4, women with severe PPH had significantly lower fibrinogen, factor V, antithrombin activity, protein C antigen, prolonged prothrombin time, and higher D-dimer and TAT complexes than women with non-severe PPH. In multivariate analysis, from H0 to H4, fibrinogen was the only marker associated with the occurrence of severe PPH. At H0, the risk for severe PPH was 2.63-fold higher for each 1 gL(-1) decrease of fibrinogen. The negative predictive value of a fibrinogen concentration >4 gL(-1) was 79% and the positive predictive value of a concentration G sequence variant suggesting WHIM syndrome. Recurrent c.1013C > G sequence variant of the CXCR4 gene resulting in p.S338X truncation mutation of this chemokine receptor protein is first reported here. PMID- 17087744 TI - Results of the PETHEMA ALL-96 trial in elderly patients with Philadelphia chromosome-negative acute lymphoblastic leukemia. AB - BACKGROUND AND AIM: Only 20-30% of elderly patients with acute lymphoblastic leukemia (ALL) are enrolled in clinical trials because of co-morbid disorders or poor performance status. We present the results of treatment of Philadelphia chromosome-negative (Ph-) ALL patients over 55 yr treated in the PETHEMA ALL-96 trial. PATIENTS AND METHODS: From 1996 to 2006, 33 patients > or = 55 yr with Ph- ALL were included. Induction therapy was vincristine, daunorubicin, prednisone, asparaginase, and cyclophosphamide over 5 weeks. Central nervous system (CNS) prophylaxis involved triple intrathecal (IT) therapy, 14 doses over the first year. Consolidation-1 included mercaptopurine, methotrexate, teniposide and cytarabine, followed by one consolidation-2 cycle similar to the induction cycle. Maintenance consisted of mercaptopurine and methotrexate up to 2 yr in complete remission (CR) with monthly reinduction cycles (vincristine, prednisone and asparaginase) during the first year. RESULTS: Median (range) age was 65 yr (56 77). Phenotype (30 patients): early-pre-B 7, common/pre-B 18, T 5. Cytogenetics (28 patients): normal 12, complex 10, t(4;11) 2 and other 4. CR was achieved in 19/33 (57.6%) patients, early death occurred in 12 (36.4%) and 2 (6%) were resistant. Overall survival and disease-free survival probabilities (2 yr, 95% CI) were 39% (21%-57%) and 46% (22%-70%), respectively (median follow up of 24 months). Removal of asparaginase and cyclophosphamide from the induction decreased induction death (OR 0.119, CI 95% 0.022-0.637, P = 0.013) and increased survival (20% vs. 52%, P = 0.05). CONCLUSIONS: The prognosis of elderly Ph- ALL patients is poor. In this study, less intensive induction decreased toxic death, allowing delivery of planned consolidation therapy and increased survival probability. PMID- 17087746 TI - USGS directions in MODFLOW development. PMID- 17087747 TI - The practical use of simplicity in developing ground water models. AB - The advantages of starting with simple models and building complexity slowly can be significant in the development of ground water models. In many circumstances, simpler models are characterized by fewer defined parameters and shorter execution times. In this work, the number of parameters is used as the primary measure of simplicity and complexity; the advantages of shorter execution times also are considered. The ideas are presented in the context of constructing ground water models but are applicable to many fields. Simplicity first is put in perspective as part of the entire modeling process using 14 guidelines for effective model calibration. It is noted that neither very simple nor very complex models generally produce the most accurate predictions and that determining the appropriate level of complexity is an ill-defined process. It is suggested that a thorough evaluation of observation errors is essential to model development. Finally, specific ways are discussed to design useful ground water models that have fewer parameters and shorter execution times. PMID- 17087748 TI - Complexity. AB - It is difficult to define complexity in modeling. Complexity is often associated with uncertainty since modeling uncertainty is an intrinsically difficult task. However, modeling uncertainty does not require, necessarily, complex models, in the sense of a model requiring an unmanageable number of degrees of freedom to characterize the aquifer. The relationship between complexity, uncertainty, heterogeneity, and stochastic modeling is not simple. Aquifer models should be able to quantify the uncertainty of their predictions, which can be done using stochastic models that produce heterogeneous realizations of aquifer parameters. This is the type of complexity addressed in this article. PMID- 17087749 TI - The role of hand calculations in ground water flow modeling. AB - Most ground water modeling courses focus on the use of computer models and pay little or no attention to traditional analytic solutions to ground water flow problems. This shift in education seems logical. Why waste time to learn about the method of images, or why study analytic solutions to one-dimensional or radial flow problems? Computer models solve much more realistic problems and offer sophisticated graphical output, such as contour plots of potentiometric levels and ground water path lines. However, analytic solutions to elementary ground water flow problems do have something to offer over computer models: insight. For instance, an analytic one-dimensional or radial flow solution, in terms of a mathematical expression, may reveal which parameters affect the success of calibrating a computer model and what to expect when changing parameter values. Similarly, solutions for periodic forcing of one-dimensional or radial flow systems have resulted in a simple decision criterion to assess whether or not transient flow modeling is needed. Basic water balance calculations may offer a useful check on computer-generated capture zones for wellhead protection or aquifer remediation. An easily calculated "characteristic leakage length" provides critical insight into surface water and ground water interactions and flow in multi-aquifer systems. The list goes on. Familiarity with elementary analytic solutions and the capability of performing some simple hand calculations can promote appropriate (computer) modeling techniques, avoids unnecessary complexity, improves reliability, and is likely to save time and money. Training in basic hand calculations should be an important part of the curriculum of ground water modeling courses. PMID- 17087750 TI - Comparison of local grid refinement methods for MODFLOW. AB - Many ground water modeling efforts use a finite-difference method to solve the ground water flow equation, and many of these models require a relatively fine grid discretization to accurately represent the selected process in limited areas of interest. Use of a fine grid over the entire domain can be computationally prohibitive; using a variably spaced grid can lead to cells with a large aspect ratio and refinement in areas where detail is not needed. One solution is to use local-grid refinement (LGR) whereby the grid is only refined in the area of interest. This work reviews some LGR methods and identifies advantages and drawbacks in test cases using MODFLOW-2000. The first test case is two dimensional and heterogeneous; the second is three dimensional and includes interaction with a meandering river. Results include simulations using a uniform fine grid, a variably spaced grid, a traditional method of LGR without feedback, and a new shared node method with feedback. Discrepancies from the solution obtained with the uniform fine grid are investigated. For the models tested, the traditional one-way coupled approaches produced discrepancies in head up to 6.8% and discrepancies in cell-to-cell fluxes up to 7.1%, while the new method has head and cell-to-cell flux discrepancies of 0.089% and 0.14%, respectively. Additional results highlight the accuracy, flexibility, and CPU time trade-off of these methods and demonstrate how the new method can be successfully implemented to model surface water-ground water interactions. PMID- 17087751 TI - Grid cell distortion and MODFLOW's integrated finite-difference numerical solution. AB - The ground water flow model MODFLOW inherently implements a nongeneralized integrated finite-difference (IFD) numerical scheme. The IFD numerical scheme allows for construction of finite-difference model grids with curvilinear (piecewise linear) rows. The resulting grid comprises model cells in the shape of trapezoids and is distorted in comparison to a traditional MODFLOW finite difference grid. A version of MODFLOW-88 (herein referred to as MODFLOW IFD) with the code adapted to make the one-dimensional DELR and DELC arrays two dimensional, so that equivalent conductance between distorted grid cells can be calculated, is described. MODFLOW IFD is used to inspect the sensitivity of the numerical head and velocity solutions to the level of distortion in trapezoidal grid cells within a converging radial flow domain. A test problem designed for the analysis implements a grid oriented such that flow is parallel to columns with converging widths. The sensitivity analysis demonstrates MODFLOW IFD's capacity to numerically derive a head solution and resulting intercell volumetric flow when the internal calculation of equivalent conductance accounts for the distortion of the grid cells. The sensitivity of the velocity solution to grid cell distortion indicates criteria for distorted grid design. In the radial flow test problem described, the numerical head solution is not sensitive to grid cell distortion. The accuracy of the velocity solution is sensitive to cell distortion with error <1% if the angle between the nonparallel sides of trapezoidal cells is <12.5 degrees. The error of the velocity solution is related to the degree to which the spatial discretization of a curve is approximated with piecewise linear segments. Curvilinear finite-difference grid construction adds versatility to spatial discretization of the flow domain. MODFLOW-88's inherent IFD numerical scheme and the test problem results imply that more recent versions of MODFLOW 2000, with minor modifications, have the potential to make use of a curvilinear grid. PMID- 17087752 TI - Positive solution of two-dimensional solute transport in heterogeneous aquifers. AB - The transport of contaminants in aquifers is usually represented by a convection dispersion equation. There are several well-known problems of oscillation and artificial dispersion that affect the numerical solution of this equation. For example, several studies have shown that standard treatment of the cross dispersion terms always leads to a negative concentration. It is also well known that the numerical solution of the convective term is affected by spurious oscillations or substantial numerical dispersion. These difficulties are especially significant for solute transport in nonuniform flow in heterogeneous aquifers. For the case of coupled reactive-transport models, even small negative concentration values can become amplified through nonlinear reaction source/sink terms and thus result in physically erroneous and unstable results. This paper includes a brief discussion about how nonpositive concentrations arise from numerical solution of the convection and cross-dispersion terms. We demonstrate the effectiveness of directional splitting with one-dimensional flux limiters for the convection term. Also, a new numerical scheme for the dispersion term that preserves positivity is presented. The results of the proposed convection scheme and the solution given by the new method to compute dispersion are compared with standard numerical methods as used in MT3DMS. PMID- 17087753 TI - An investigation of numerical grid effects in parameter estimation. AB - Modern ground water characterization and remediation projects routinely require calibration and inverse analysis of large three-dimensional numerical models of complex hydrogeological systems. Hydrogeologic complexity can be prompted by various aquifer characteristics including complicated spatial hydrostratigraphy and aquifer recharge from infiltration through an unsaturated zone. To keep the numerical models computationally efficient, compromises are frequently made in the model development, particularly, about resolution of the computational grid and numerical representation of the governing flow equation. The compromise is required so that the model can be used in calibration, parameter estimation, performance assessment, and analysis of sensitivity and uncertainty in model predictions. However, grid properties and resolution as well as applied computational schemes can have large effects on forward-model predictions and on inverse parameter estimates. We investigate these effects for a series of one- and two-dimensional synthetic cases representing saturated and variably saturated flow problems. We show that "conformable" grids, despite neglecting terms in the numerical formulation, can lead to accurate solutions of problems with complex hydrostratigraphy. Our analysis also demonstrates that, despite slower computer run times and higher memory requirements for a given problem size, the control volume finite-element method showed an advantage over finite-difference techniques in accuracy of parameter estimation for a given grid resolution for most of the test problems. PMID- 17087754 TI - Stratigraphic control of flow and transport characteristics. AB - Ground water flow and travel time are dependent on stratigraphic architecture, which is governed by competing processes that control the spatial and temporal distribution of accommodation and sediment supply. Accommodation is the amount of space in which sediment may accumulate as defined by the difference between the energy gradient and the topographic surface. The temporal and spatial distribution of accommodation is affected by processes that change the distribution of energy (e.g., sea level or subsidence). Fluvial stratigraphic units, generated by FLUVSIM (a stratigraphic simulator based on accommodation and sediment supply), with varying magnitudes and causes of accommodation, were incorporated into a hydraulic regime using MODFLOW (a ground water flow simulator), and particles were tracked using MODPATH (a particle-tracking algorithm). These experiments illustrate that the dominant type of accommodation process influences the degree of continuity of stratigraphic units and thus affects ground water flow and transport. When the hydraulic gradient is parallel to the axis of the fluvial system in the depositional environment, shorter travel times occur in low-total accommodation environments and longer travel times in high-total accommodation environments. Given the same total accommodation, travel times are longer when sea-level change is the dominant process than those in systems dominated by subsidence. PMID- 17087755 TI - Karst spring responses examined by process-based modeling. AB - Ground water in karst terrains is highly vulnerable to contamination due to the rapid transport of contaminants through the highly conductive conduit system. For contamination risk assessment purposes, information about hydraulic and geometric characteristics of the conduits and their hydraulic interaction with the fissured porous rock is an important prerequisite. The relationship between aquifer characteristics and short-term responses to recharge events of both spring discharge and physicochemical parameters of the discharged water was examined using a process-based flow and transport model. In the respective software, a pipe-network model, representing fast conduit flow, is coupled to MODFLOW, which simulates flow in the fissured porous rock. This hybrid flow model was extended to include modules simulating heat and reactive solute transport in conduits. The application of this modeling tool demonstrates that variations of physicochemical parameters, such as solute concentration and water temperature, depend to a large extent on the intensity and duration of recharge events and provide information about the structure and geometry of the conduit system as well as about the interaction between conduits and fissured porous rock. Moreover, the responses of solute concentration and temperature of spring discharge appear to reflect different processes, thus complementing each other in the aquifer characterization. PMID- 17087756 TI - River-aquifer interactions, geologic heterogeneity, and low-flow management. AB - Low river flows are commonly controlled by river-aquifer exchange, the magnitude of which is governed by hydraulic properties of both aquifer and aquitard materials beneath the river. Low flows are often important ecologically. Numerical simulations were used to assess how textural heterogeneity of an alluvial system influences river seepage and low flows. The Cosumnes River in California was used as a test case. Declining fall flows in the Cosumnes River have threatened Chinook salmon runs. A ground water-surface water model for the lower river basin was developed, which incorporates detailed geostatistical simulations of aquifer heterogeneity. Six different realizations of heterogeneity and a homogenous model were run for a 3-year period. Net annual seepage from the river was found to be similar among the models. However, spatial distribution of seepage along the channel, water table configuration and the level of local connection, and disconnection between the river and aquifer showed strong variations among the different heterogeneous models. Most importantly, the heterogeneous models suggest that river seepage losses can be reduced by local reconnections, even when the regional water table remains well below the riverbed. The percentage of river channel responsible for 50% of total river seepage ranged from 10% to 26% in the heterogeneous models as opposed to 23% in the homogeneous model. Differences in seepage between the models resulted in up to 13 d difference in the number of days the river was open for salmon migration during the critical fall months in one given year. PMID- 17087757 TI - Transition probability/Markov chain analyses of DNAPL source zones and plumes. AB - At sites where a dense nonaqueous phase liquid (DNAPL) was spilled or released into the subsurface, estimates of the mass of DNAPL contained in the subsurface from core or monitoring well data, either in the nonaqueous or aqueous phase, can be highly uncertain because of the erratic distribution of the DNAPL due to geologic heterogeneity. In this paper, a multiphase compositional model is applied to simulate, in detail, the DNAPL saturations and aqueous-phase plume migration in a highly characterized, heterogeneous glaciofluvial aquifer, the permeability and porosity data of which were collected by researchers at the University of Tubingen, Germany. The DNAPL saturation distribution and the aqueous-phase contaminant mole fractions are then reconstructed by sampling the data from the forward simulation results using two alternate approaches, each with different degrees of sampling conditioning. To reconstruct the DNAPL source zone architecture, the aqueous-phase plume configuration, and the contaminant mass in each phase, one method employs the novel transition probability/Markov chain approach (TP/MC), while the other involves a traditional variogram analysis of the sampled data followed by ordinary kriging. The TP/MC method is typically used for facies and/or hydraulic conductivity reconstruction, but here we explore the applicability of the TP/MC method for the reconstruction of DNAPL source zones and aqueous-phase plumes. The reconstructed geometry of the DNAPL source zone, the dissolved contaminant plume, and the estimated mass in each phase are compared using the two different geostatistical modeling approaches and for various degrees of data sampling from the results of the forward simulation. It is demonstrated that the TP/MC modeling technique is robust and accurate and is a preferable alternative compared to ordinary kriging for the reconstruction of DNAPL saturation patterns and dissolved-phase contaminant plumes. PMID- 17087758 TI - Reducing long-term remedial costs by transport modeling optimization. AB - The Department of Defense (DoD) Environmental Security Technology Certification Program and the Environmental Protection Agency sponsored a project to evaluate the benefits and utility of contaminant transport simulation-optimization algorithms against traditional (trial and error) modeling approaches. Three pump and-treat facilities operated by the DoD were selected for inclusion in the project. Three optimization formulations were developed for each facility and solved independently by three modeling teams (two using simulation-optimization algorithms and one applying trial-and-error methods). The results clearly indicate that simulation-optimization methods are able to search a wider range of well locations and flow rates and identify better solutions than current trial and-error approaches. The solutions found were 5% to 50% better than those obtained using trial-and-error (measured using optimal objective function values), with an average improvement of approximately 20%. This translated into potential savings ranging from 600,000 dollars to 10,000,000 dollars for the three sites. In nearly all cases, the cost savings easily outweighed the costs of the optimization. To reduce computational requirements, in some cases the simulation-optimization groups applied multiple mathematical algorithms, solved a series of modified subproblems, and/or fit "meta-models" such as neural networks or regression models to replace time-consuming simulation models in the optimization algorithm. The optimal solutions did not account for the uncertainties inherent in the modeling process. This project illustrates that transport simulation-optimization techniques are practical for real problems. However, applying the techniques in an efficient manner requires expertise and should involve iterative modification to the formulations based on interim results. PMID- 17087759 TI - Challenges to calibration: Facing an increasingly critical environment. PMID- 17087760 TI - Aurora kinase is required for chromosome segregation in tobacco BY-2 cells. AB - Post-translational modifications of core histone tails play crucial roles in chromatin structure and function. Although phosphorylation of Ser10 and Ser28 (H3S10ph and H3S28ph) of histone H3 is ubiquitous among eukaryotes, the phosphorylation mechanism during the cell cycle remains unclear. In the present study, H3S10ph and H3S28ph in tobacco BY-2 cells were observed in the pericentromeric regions during mitosis. Moreover, the Aurora kinase inhibitor Hesperadin inhibited the kinase activity of Arabidopsis thaliana Aurora kinase 3 (AtAUR3) in phosphorylating both Ser10 and Ser28 of histone H3 in vitro. Consistently, Hesperadin inhibited both H3S10ph and H3S28ph during mitosis in BY 2 cells. These results indicate that plant Aurora kinases phosphorylate not only Ser10, but also Ser28 of histone H3 in vivo. Hesperadin treatment increased the ratio of metaphase cells, while the ratio of anaphase/telophase cells decreased, although the mitotic index was not affected in Hesperadin-treated cells. These results suggest that Hesperadin induces delayed transition from metaphase to anaphase, and early exit from mitosis after chromosome segregation. In addition, micronuclei were observed frequently and lagging chromosomes, caused by the delay and failure of sister chromatid separation, were observed at anaphase and telophase in Hesperadin-treated BY-2 cells. The data obtained here suggest that plant Aurora kinases and H3S10ph/H3S28ph may have a role in chromosome segregation and metaphase/anaphase transition. PMID- 17087761 TI - Time-lapse analysis of stem-cell divisions in the Arabidopsis thaliana root meristem. AB - In the Arabidopsis root, asymmetric stem-cell divisions produce daughters that form the different root cell types. Here we report the establishment of a confocal tracking system that allows the analysis of numbers and orientations of cell divisions in root stem cells. The system provides direct evidence that stem cells have lower division rates than cells in the proximal meristem. It also allows tracking of cell division timing, which we have used to analyse the synchronization of root cap divisions. Finally, it gives new insights into lateral root cap formation: epidermal stem-cell daughters can rotate the orientation of the division plane like the stem cell. PMID- 17087770 TI - Apoptosis in yeast--mechanisms and benefits to a unicellular organism. AB - Initial observations that the budding yeast Saccharomyces cerevisiae can be induced to undergo a form of cell death exhibiting typical markers of apoptosis has led to the emergence of a thriving new field of research. Since this discovery, a number of conserved pro- and antiapoptotic proteins have been identified in yeast. Indeed, early experiments have successfully validated yeasts as a powerful genetic tool with which to investigate mechanisms of apoptosis. However, we still have little understanding as to why programmes of cell suicide exist in unicellular organisms and how they may be benefit such organisms. Recent research has begun to elucidate pathways that regulate yeast apoptosis in response to environmental stimuli. These reports strengthen the idea that physiologically relevant mechanisms of programmed cell death are present, and that these function as important regulators of yeast cell populations. PMID- 17087771 TI - Control of fatty acid desaturation: a mechanism conserved from bacteria to humans. AB - Unsaturated fatty acids (UFAs) have profound effects on the fluidity and function of biological membranes. Microorganisms, plants and animals regulate the synthesis of UFAs during changing environmental conditions as well as in response to nutrients. UFAs homeostasis in many organisms is achieved by feedback regulation of fatty acid desaturase gene transcription through signalling pathways that are governed by sensors embedded in cellular membranes. Here, we review recently discovered components of the regulatory machinery governing the transcription of fatty acid desaturases in bacteria, yeasts and animals that indicate an ancient role of transmembrane signalling mechanisms and integrate membrane composition with lipid biosynthesis. PMID- 17087772 TI - Novel DNA binding protein SarZ contributes to virulence in Staphylococcus aureus. AB - We previously reported that the cvfA gene is a virulence regulatory gene in Staphylococcus aureus. Here, we identified a novel gene named sarZ that acts as a multicopy suppressor of decreased haemolysin production in the cvfA deletion mutant. The amount of sarZ transcripts was decreased in the cvfA mutant. The sarZ deletion mutant produced less haemolysin and attenuated virulence in a silkworm infection model and a mouse-infection model. The amino acid sequence of the sarZ gene product had 19% identity with the transcription factor MarR in Escherichia coli, and the internal region contained a winged helix-turn-helix motif (wHTH), a known DNA binding domain. Purified recombinant SarZ protein had binding affinity for the promoter region of the hla gene that encodes alpha-haemolysin. SarZ mutant proteins with an amino acid substitution in the N-terminal region or in the wHTH motif had significantly decreased DNA binding. The mutated sarZ genes encoding SarZ mutant proteins with a low affinity for DNA did not complement the decreased haemolysin production or the attenuated killing ability against silkworms in the sarZ mutant. These results suggest that the DNA binding activity of the SarZ protein is required for virulence in S. aureus. PMID- 17087773 TI - IHF-dependent activation of P1 plasmid origin by dnaA. AB - In bacteria, many DNA-protein interactions that initiate transcription, replication and recombination require the mediation of DNA architectural proteins such as IHF and HU. For replication initiation, plasmid P1 requires three origin binding proteins: the architectural protein HU, a plasmid-specific initiator, RepA, and the Escherichia coli chromosomal initiator, DnaA. The two initiators bind in the origin of replication to multiple sites, called iterons and DnaA boxes respectively. We show here that all five known DnaA boxes can be deleted from the plasmid origin provided the origin is extended by about 120 bp. The additional DNA provides an IHF site and most likely a weak DnaA binding site, because replacing the putative site with an authentic DnaA box enhanced plasmid replication in an IHF-dependent manner. IHF most likely brings about interactions between distally bound DnaA and RepA by bending the intervening DNA. The role of IHF in activating P1 origin by allowing DnaA binding to a weak site is reminiscent of the role the protein plays in initiating the host chromosomal replication. PMID- 17087775 TI - The symbiosis regulator rscS controls the syp gene locus, biofilm formation and symbiotic aggregation by Vibrio fischeri. AB - Successful colonization of a eukaryotic host by a microbe involves complex microbe-microbe and microbe-host interactions. Previously, we identified in Vibrio fischeri a putative sensor kinase, RscS, required for initiating symbiotic colonization of its squid host Euprymna scolopes. Here, we analysed the role of rscS by isolating an allele, rscS1, with increased activity. Multicopy rscS1 activated transcription of genes within the recently identified symbiosis polysaccharide (syp) cluster. Wild-type cells carrying rscS1 induced aggregation phenotypes in culture, including the formation of pellicles and wrinkled colonies, in a syp-dependent manner. Colonies formed by rscSl-expressing cells produced a matrix not found in control colonies and largely lost in an rscSl expressing sypN mutant. Finally, multicopy rscS1 provided a colonization advantage over control cells and substantially enhanced the ability of wild-type cells to aggregate on the surface of the symbiotic organ of E. scolopes; this latter phenotype similarly depended upon an intact syp locus. These results suggest that transcription induced by RscS-mediated signal transduction plays, a key role in colonization at the aggregation stage by modifying the cell surface and increasing the ability of the cells to adhere to one another and/or to squid secreted mucus. PMID- 17087776 TI - RNase R degrades non-stop mRNAs selectively in an SmpB-tmRNA-dependent manner. AB - The SmpB-tmRNA-mediated trans-translation system has two well-established activities: rescuing ribosomes stalled on aberrant mRNAs and marking the associated protein fragments for proteolysis. Although the causative non-stop mRNAs are known to be degraded, little is known about the enabling mechanism or the RNases involved in their disposal. We report that Escherichia coli has an enabling mechanism that requires RNase R activity and is dependent on the presence of SmpB protein and tmRNA, suggesting a requirement for active transtranslation in facilitating RNase R engagement and promoting non-stop mRNA decay. Interestingly, this selective transcript degradation by RNase R targets aberrant (non-stop and multiple-rare-codon containing) mRNAs and does not affect the decay of related messages containing in-frame stop codons. Most surprisingly, RNase II and PNPase do not play a significant role in tmRNA-facilitated disposal of aberrant mRNAs. These findings demonstrate that RNase R is a crucial component of the trans-translation-mediated non-stop mRNA decay process, thus providing a requisite activity well suited to complement the ribosome rescue and protein tagging functions of this unique quality control system. PMID- 17087777 TI - Aspirin reduces anticardiolipin antibodies in patients with coronary artery disease. AB - BACKGROUND: Anticardiolipin antibodies (aCL) have been found to be elevated in patients with coronary artery disease (CAD) and have been associated with an adverse outcome owing to their prothrombotic activity. The aim of this study was to investigate the effect of aspirin treatment on aCL levels in patients with chronic CAD. MATERIALS AND METHODS: Forty patients with chronic CAD scheduled for elective coronary artery bypass graft surgery (CABG) and 40 healthy controls participated in the study. Patients were treated with 300 mg of aspirin once daily (o.d.) for the first 12 days and placebo for the following 12 days before CABG in a double-blind, cross-over trial. Immunoglobulin (Ig) G-, IgM-, IgA-aCL and C-reactive protein (CRP) levels were measured in the controls and at the end of each treatment period in the patients with CAD. RESULTS: The IgA- and IgG-aCL levels were greater in patients with CAD than in the controls. Compared with the placebo, IgA, IgG subtypes and CRP levels were reduced after aspirin treatment (P = 0.001, P = 0.02, P = 0.04, respectively). The percentage reduction of IgA- and IgG-aCL was related to the percentage reduction of CRP after aspirin (P < 0.05). CONCLUSION: Aspirin treatment with 300 mg o.d. reduced the serum levels of IgA and IgG subtypes in patients with chronic CAD in parallel to a reduction in CRP. These findings offer an additional pathophysiological mechanism of the beneficial effects of aspirin in patients with chronic CAD. PMID- 17087778 TI - Acute effects of caffeine and tobacco on arterial function and wave travel. AB - BACKGROUND: Caffeine and tobacco consumption are risk factors for heart failure, but their effects remain controversial. It has been hypothesized that they cause alterations in arterial stiffness and arterial wave travel which may increase ventricular loading. In this study the authors examined the influence of these widely used stimulants on wave intensity and arterial stiffness parameters using carotid wave intensity analysis. MATERIALS AND METHODS: A new Doppler-based ultrasound method was used to measure the acute effects of caffeine and tobacco on wave intensity in the right common carotid artery. The measurements enabled changes in arterial stiffness parameters to be recorded. RESULTS: In 17 subjects compared with 10 controls, caffeine increased blood pressure, early systolic wave intensity and wave speed, but late-systolic wave intensity and mid-systolic reflections were unchanged. In 11 smokers studied before and after smoking one cigarette, blood pressure and arterial stiffness increased but wave intensity was unchanged. No changes were observed in the controls. CONCLUSIONS: Increased wave intensity during ejection after caffeine suggested sympathomimetic effects on the left ventricular function. Increased wave speed in the common carotid artery implied augmented central loading after caffeine, but the absence of measurable changes in local arterial stiffness in the carotid artery suggested more complex and regional effects. Cigarette smoking acutely increased local arterial stiffness in the common carotid artery. These changes can be detected using wave intensity analysis. PMID- 17087779 TI - High- but not low-dose folic acid improves endothelial function in coronary artery disease. AB - BACKGROUND: While folic acid (FA) reduces plasma homocysteine (Hcy), whether the simultaneous improvement in endothelial function is dependent on Hcy lowering per se is questionable. In the present study the relationship between FA dose, Hcy lowering and endothelial function in patients with coronary artery disease (CAD) was investigated. MATERIALS AND METHODS: Eighty-four patients with CAD received either 400 microg FA or 5 mg placebo daily for a 6-week treatment period. A further 44 patients with CAD received either 100 mg kg(-1) day(-1) of betaine or placebo for a 6-week treatment period. Flow-mediated dilatation (FMD), a measure of endothelial function, was assessed before and after the 6-week periods. Isometric tension and Western blotting were used to investigate the effect of FA on endothelial function and endothelial nitric oxide synthase (eNOS) dimerization in isolated rabbit aortic rings and cultured porcine aortic endothelial cells (PAEC), respectively. RESULTS: Both 400 micro g day(-1) and 5 mg day(-1) FA significantly increased plasma folate and decreased plasma Hcy. The FMD improved significantly after 6 weeks' treatment of 5 mg day(-1) FA but did not correlate with the reduction in Hcy. There was no change in FMD in either the 400 micro g FA or placebo group. In a subgroup analysis of 11 patients in the betaine group, despite a reduced Hcy, a significant impairment in FMD was observed. In the in vitro studies FA, but not betaine, reversed methionine-induced endothelial dysfunction. Moreover, the FA promoted eNOS dimerization in cultured PAEC. CONCLUSIONS: These data suggest that FA dose-dependently improves endothelial function in CAD via a mechanism independently of Hcy lowering. It may involve promotion of eNOS dimerization. PMID- 17087780 TI - Inflammation affects lipid metabolism during recovery from hyperinsulinaemia. AB - BACKGROUND: Inflammation is an established contributor in atherosclerosis and several other common diseases including diabetes. Therefore the study was to investigate how inflammatory factors affect lipid metabolism during recovery from hyperinsulinaemia in healthy individuals. MATERIALS AND METHODS: A total of 22 healthy subjects [aged 27.7 +/- 1.8 years; body mass index (BMI) 24.1 +/- 0.8 kg m(-2)] participated in the study. After a 4-h euglycaemic hyperinsulinaemia (55.9 +/- 2.2 mU L(-1)) insulin infusion was stopped and baseline blood samples were taken. Glucose infusion at a decreasing rate continued for 120 min to maintain euglycaemia throughout the study. RESULTS: The free fatty acid (FFA) concentration at the 120-min time-point was associated with baseline alpha-1-acid glycoprotein (A1GP) (r = 0.57, P < 0.01), C-reactive protein (CRP) (r = 0.54, P < 0.02) and serum amyloid A (r = 0.53, P < 0.02); in total they accounted for 54% of the variation in FFA concentration at the 120-min time-point. Baseline A1GP was also associated with the triglyceride concentration at the 120-min time-point (r = 0.66, P < 0.001). Insulin sensitivity was the most important factor associated with glucose disposal at the 120-min time-point, thus explaining 30% of the variation (P < 0.01). Interleukin-6 (positive correlation) and fibrinogen (negative correlation) increased the proportion to 48% (P < 0.01). There was no significant change in the most acute-phase proteins between baseline and the 120 min time-point. CONCLUSION: Inflammation is an important contributor to lipid and glucose metabolism during recovery from hyperinsulinaemia. PMID- 17087781 TI - Identification and characterization of novel low-density lipoprotein receptor mutations of familial hypercholesterolaemia patients in Taiwan. AB - BACKGROUND: Familial hypercholesterolaemia (FH) is an autosomal dominant disease associated with a very high risk of coronary vascular disease. The study objective was to identify patients with FH in Taiwan and characterize novel mutations. MATERIALS AND METHODS: Fifty-one patients with suspected FH living in Taiwan were screened for mutations in both the low-density lipoprotein (LDL) receptor and the apolipoprotein (apoB) genes using the multiplex polymerase chain reaction and exon-by-exon DNA sequencing technique. Functional consequences on LDL receptor activity were characterized in vitro for novel mutations and family pedigree was also analyzed. RESULTS: Thirteen different functional mutations in the LDL receptor gene and one mutation in the apoB gene were found in 21 patients. Among the 13 mutations in the LDL receptor gene, 10 were single-point missense mutations, one was a two-point mutation in the same allele, one was a non-sense mutation and one was a frame-shift mutation. There were three novel mutations, including two missense mutations (M510K and W512R) and one frame-shift mutation (1953 delTA mutation). The characterization of missense M510K retained 36.2% of the activity of the normal receptor. Conversely, frame-shift 1953 delTA and missense W512R led to defective proteins, with only 0-6% of normal receptor activity. CONCLUSIONS: The study identified 13 LDL receptor gene mutations and characterized three novel mutations causing FH in Taiwan. This facilitated a better understanding of FH among the Chinese population and may enable diagnosis of FH at the molecular level at a presymptomatic, early age. PMID- 17087782 TI - ApoA-I mutants V156K and R173C promote anti-inflammatory function and antioxidant activities. AB - BACKGROUND: Two mutants of apolipoprotein (apo) A-I, V156K and A158E, showed markedly different structural and functional properties in lipid-free and lipid bound states in the authors' earlier report. The physiological activities of these mutants were compared with the wild-type (WT) and R173C mutant using in vitro and in vivo experiments. MATERIALS AND METHODS: A reconstituted high density lipoprotein (rHDL) with palmitoyloleoyl phosphatidylcholine (POPC), combined with each of the apoA-I variants, was injected into the tail-veins of hypercholesterolaemic mice (C57BL6/J), which had been fed a high cholesterol and high fat (HCHF; 0.5% cholesterol, 15% lard, 0.1% sodium cholate) diet for 23 weeks, once at 0 h and then every 24 h, at a dosage of 30 mg apoA-I kg(-1) of body-weight. RESULTS: The V156K-rHDL and R173C-rHDL exhibited significantly stronger anti-oxidant activity against copper-mediated low-density lipoprotein (LDL) oxidation than did A158E in an apolipoprotein state. The mice injected with WT-rHDL or A158E-rHDL showed abrupt increases in total cholesterol concentrations (47% and 38%, respectively) as compared with the levels before injection, whereas the mice injected with V156K-rHDL and R173C-rHDL did not. Injection with V156K rHDL improved serum lipids and anti-oxidative activities compared with the injection of WT-rHDL. Injection of WT-rHDL or A158E-rHDL increased serum interleukin-6 (IL-6) to 90-110 pg mL(-1), whereas the injection of V156K-rHDL or R173C-rHDL increased serum IL-6 to 17-25 pg mL(-1) only. CONCLUSION: The V156K rHDL and R173C-rHDL displayed potent beneficial effects, including anti-oxidant and anti-inflammatory activity from both in vitro and in vivo evaluations, whereas the WT-rHDL and A158E-rHDL did not. PMID- 17087783 TI - Women with polycystic ovary syndrome are sensitive to the TNF-alpha-lowering effect of glucose-induced hyperinsulinaemia. AB - BACKGROUND: Restoration of near-euglycaemia by intensive insulin therapy results in decreased serum levels of inflammatory mediators. The authors investigated whether the anti-inflammatory effect of insulin was independent of its glucose lowering action and if this effect was intact in insulin-resistant women with the polycystic ovary syndrome (PCOS) characterized by low-grade chronic inflammation. MATERIALS AND METHODS: Blood was drawn on the third and sixth days after progestin-induced withdrawal bleeding in 20 young non-diabetic women with PCOS and once between the third and sixth days of the menstrual cycle in 21 age matched lean healthy control women during a 75-g oral glucose tolerance test (oGTT). Serum insulin, glucose and tumour necrosis factor alpha (TNF-alpha) concentrations were measured after 0, 30, 60, 90 and 120 min. RESULTS: The increase in insulin and glucose concentrations during the oGTT was significantly more pronounced in patients with PCOS (one patient with impaired fasting glucose, one patient with impaired glucose tolerance, three patients with both) compared with healthy controls. The TNF-alpha serum concentrations decreased in patients with PCOS (mean of both days, P = 0.004). In patients and in controls, there was an inverse correlation between the serum concentrations of insulin and of TNF alpha during oGTT (for patients, a mean of both days, P = 0.009; for controls, P = 0.047), but not between the serum concentrations of glucose and TNF-alpha. CONCLUSIONS: The decrease in TNF-alpha concentrations during oGTT and the inverse correlation between endogenous hyperinsulinaemia and serum TNF-alpha concentrations suggested an anti-inflammatory effect of moderately-high insulin concentrations. This occurred despite the presence of moderate hyperglycaemia. These findings also demonstrated a preserved responsiveness of inflammatory mediators to insulin in PCOS. PMID- 17087784 TI - Immunolocalization of duodenal cytochrome B: a relationship with circulating markers of iron status. AB - BACKGROUND: The brush border ferric reductase (Dcytb) is critical for the absorption of dietary iron and appears to be expressed on the duodenal enterocyte brush border. The Dcytb expression is increased in severe iron-deficient anaemia, but the situation in a more typical mild iron deficiency is unclear. This study investigated Dcytb expression in patients with normal iron status or mild iron deficiency and its relationships with enterocyte iron status. MATERIALS AND METHODS: Duodenal biopsy specimens and blood samples were obtained from 32 patients undergoing routine upper gastrointestinal endoscopy. Twenty-three specimens (six iron-deficient and 17 iron-replete) were processed for light microscopy (LM) and for immunohistochemistry with antibodies against Dcytb and heavy/light chain ferritin subunits. The nine remaining biopsies (three iron deficient and six iron-replete) were processed for electron microscopy (EM). Immunolocalization of Dcytb and intracellular ferritin was performed with appropriate primary antibodies followed by 10-nm gold conjugate labels. RESULTS: The LM process showed a strong negative correlation between immunolabelling intensity of Dcytb on the enterocyte brush border and serum iron saturation (P < 0.001), but only a weak negative correlation between this antigen and haemoglobin (P = 0.08) or serum ferritin concentrations (P = 0.4). EM confirmed anti-Dcytb preferential labelling of microvilli rather than enterocyte cytoplasm (P = 0.001), but preferential antiferritin labelling of cytoplasm (P < 0.02). There was no correlation with enterocyte cytoplasmic ferritin labelling (i.e. enterocyte iron status and Dcytb expression). CONCLUSIONS: Enterocyte Dcytb brush border expression is increased even in mild iron deficiency and may be related to serum iron saturation. The lack of correlation with enterocyte ferritin expression deserves further study with direct measurement of intracellular iron. PMID- 17087785 TI - AMP challenge induces a decrease in FE(NO) in asthmatic subjects modulated by nedocromil. AB - BACKGROUND: Allergen challenge results in an immediate reduction in exhaled nitric oxide (FE(NO)) followed by a long-term increase. To study mast cell activation in relation to nitric oxide (NO), the study investigated the effect of inhaled adenosine monophosphate (AMP) as a mast cell activator and mast cell stabilizer - nedocromil sodium - on FE(NO). The NO synthase (NOS) iso-enzyme involved was studied by the NOS inhibitor aminoguanidine. MATERIALS AND METHODS: A double-blind, placebo-controlled, cross-over study was performed in two parts. Part I: eight atopic asthmatic subjects inhaled nedocromil or placebo before the AMP challenge. Spirometry and FE(NO) were measured at intervals over a 24-h period. Part II: seven subjects inhaled aminoguanidine before an identical protocol was used, as in Part I. RESULTS: Part I: AMP challenge caused a significant decrease from baseline FE(NO)[placebo, 28.9 (20.3-37.4)%, P < 0.002 and nedocromil, 20.9 (8.2-33.6)%, P < 0.01]. Nedocromil gave partial protection against this decrease in FE(NO). The time-FE(NO) curve (AUC(0-24)) differed significantly between nedocromil and placebo: 2.7% (-3.6 to -9) vs. -6.6% (-12 to -1.3) FE(NO) changes h(-1), P < 0.002, respectively. Nedocromil protected against AMP-induced bronchoconstriction (AMP PC(20)) [nedocromil 182 (72.5-291) mg mL(-1) vs. placebo 21.7 (10.7-33) mg mL(-1), P < 0.002]. Part II: nebulized aminoguanidine resulted in a significant reduction in FE(NO) from baseline and was greater than after AMP alone (P = 0.006). Nedocromil increased AMP PC(20), but no longer protected against the late decrease in FE(NO). CONCLUSIONS: The AMP challenge caused a reduction in FE(NO) as a result of prior treatment with nedocromil. Aminoguanidine abolished the nedocromil-induced protection on the late reduction in FE(NO), but not on AMP PC(20). Inducible NOS was implicated in the late FE(NO) decrease after the AMP challenge. PMID- 17087786 TI - Psychiatry and the humanities. PMID- 17087787 TI - Antidepressants and sexual dysfunction. AB - OBJECTIVE: Many patients with depression suffer from sexual dysfunction and sexual dysfunction is a recognized side-effect of antidepressants. The aim of this review was to examine the prevalence of psychosexual dysfunction associated with antidepressants, and to review treatment options which are specific to the affected component of sexual functioning and antidepressants. METHOD: Comprehensive literature review using Medline and Cochrane databases. RESULTS: Up to 70% of patients with depression may have sexual dysfunction. Tricyclic antidepressants, selective-serotonin reuptake inhibitors and venlafaxine are most and the non-serotonergic antidepressants and duloxetine least likely to produce sexual dysfunction. Pharmacological treatment options include antidepressants less likely associated or 'antidotes' to reverse sexual dysfunction. CONCLUSION: Sexual dysfunction may be a preventable or treatable side-effect of antidepressants. Patients need routinely to be asked about sexual function to identify problems early. If sexual dysfunction is ignored it may maintain the depression, compromise treatment outcome and lead to non-compliance. PMID- 17087788 TI - The criterion validity of the Geriatric Depression Scale: a systematic review. AB - OBJECTIVE: The objective was to provide a systematic review of the screening accuracy of both versions of the Geriatric Depressions Scale (GDS-30, GDS-15). METHOD: An electronic search was performed by using Medline, Embase, Cinahl, Psyndex and the Cochrane library. The selection and examination of papers were performed by two reviewers independently. RESULTS: Among the 42 papers included, important methodological aspects such as sampling methods or blinding of research workers often were not reported. For both GDS versions, similar validity indices were found (GDS-30: sensitivity 0.753, specificity 0.770; GDS-15: sensitivity 0.805, specificity 0.750). Using comparative studies based on the identical samples, both GDS versions showed significantly better validity indices than the 'Yale-1-question' screen, but were similar to the 'Center for Epidemiological Studies Depression scale' (CES-D). CONCLUSION: The GDS does not show a better criterion validity than the CES-D, but methodological limitations of primary studies hamper the generalizability of pooled analyses. PMID- 17087789 TI - The persistence of depression score. AB - OBJECTIVE: To construct a score that allows prediction of major depressive episode (MDE) persistence in individuals with MDE using determinants of persistence identified in previous research. METHOD: Data were derived from 250 subjects from the general population with new MDE according to DSM-III-R. These subjects were recruited from the Netherlands Mental Health Survey and Incidence Study (NEMESIS), using the Composite International Diagnostic Interview. Determinants for persistence were transformed into a practical risk score using proportional hazards models and bootstrapping techniques. RESULTS: The risk of MDE persistence after 12 months was 23%. The score comprised measures of physical illness, social support, depression severity and recurrency, and duration of previous episodes. With increasing categories of these measures, the predicted risks increased from 7 to 40%. CONCLUSION: We constructed the Persistence of Depression Score (PDS) showing reasonable performance. The PDS could be of importance in clinical practice to support treatment decisions. PMID- 17087790 TI - Personality traits in early phases of panic disorder: implications on the presence of agoraphobia, clinical severity and short-term outcome. AB - OBJECTIVE: To explore the relations between personality traits using the Big Five model and presence of agoraphobia, clinical severity and short-term outcome in an unbiased clinical sample of never-treated panic disorder patients. METHOD: Panic disorder (PD) patients (n = 103) in the first stages of their illness were evaluated using the Neuroticism-Extraversion-Openness Five Factor Inventory of Personality (NEO-FFI) and were compared with a sample of healthy subjects. Severity was assessed by the Panic Disorder Severity Scale and the Clinical Global Impression Scales. Patients were evaluated after 8 weeks of naturalistic pharmacologic treatment with Selective Serotonin Reuptake Inhibitors. RESULTS: Panic disorder patients show more neuroticism than healthy subjects. Patients suffering from agoraphobia are more introverted than controls. Extraversion, in addition to gender and distress, during panic attacks allows to correctly classifying 72% of the cases of agoraphobia. CONCLUSION: Low scores in extraversion contribute to explain the presence of agoraphobia in panic disorder. Personality traits are neither related to clinical severity nor to short-term response to pharmacological treatment. PMID- 17087791 TI - Clinical outcome in bipolar disorder in a community-based follow-up study in Butajira, Ethiopia. AB - OBJECTIVE: To determine the clinical outcome of bipolar disorder in a developing country setup. METHOD: After assessing 68 378 individuals, aged 15-49 years, in a double-sampling design in a rural community in Ethiopia, 312 patients with bipolar disorder were prospectively monitored with symptom rating scales and clinically for an average of 2.5 years. RESULTS: Overall, 65.9% of the cohort experienced a relapse--47.8% manic, 44.3% depressive and 7.7% mixed episodes--and 31.1% had persistent illness. Female gender predicted depressive relapse, while male gender predicted manic relapse. Being on psychotropic medication was associated with remission. CONCLUSION: This large community-based study confirms the relapsing nature of bipolar disorder and a tendency for chronicity. This may be partly because of lack of appropriate interventions in this setting; however, it may also indicate the underlying severity of the disorder irrespective of setting. PMID- 17087792 TI - C957T DRD2 polymorphism is associated with schizophrenia in Spanish patients. AB - OBJECTIVE: The objective was to confirm whether a homozygous genotype for the C957 allele of the C957T DRD2 gene single nucleotide polymorphism (SNP) is associated with schizophrenia in an independent study population. METHOD: We examined the genotypic distribution of this SNP in a set of clinically ascertained schizophrenic patients (n = 131) and age-matched control subjects (n = 364). Individuals were genotyped using automated analysis of fluorescently labeled PCR products. RESULTS: The distribution of grouped genotypes for the C957T DRD2 SNP (CC vs. CT, TT) showed that C homozygote genotype was over represented in our patient sample when compared with control subjects. This difference reaches the statistical significance (chi(2) = 7.0; df = 1; P = 0.008; OR = 2.05; % CI 1.2-3.4). CONCLUSION: The findings of this study provide additional evidence that genetic variation at the DRD2 gene plays an important role in the vulnerability to schizophrenia. PMID- 17087793 TI - Friedrich Nietzsche's mental illness--general paralysis of the insane vs. frontotemporal dementia. AB - OBJECTIVE: For a long time it was thought that Nietzsche suffered from general paralysis of the insane (GPI). However, this diagnosis has been questioned recently, and alternative diagnoses have been proposed. METHOD: We have charted Friedrich Nietzsche's final fatal illness, and viewed the differential diagnosis in the light of recent neurological understandings of dementia syndromes. RESULTS: It is unclear that Nietzsche ever had syphilis. He lacked progressive motor and other neurological features of a progressive syphilitic central nervous system (CNS) infection and lived at least 12 years following the onset of his CNS signs, which would be extremely rare for patients with untreated GPI. Finally, his flourish of productivity in 1888 would be quite uncharacteristic of GPI, but in keeping with reports of burgeoning creativity at some point in the progression of frontotemporal dementia (FTD). CONCLUSION: We suggest that Nietzsche did not have GPI, but died from a chronic dementia, namely FTD. PMID- 17087795 TI - Waggish excerpts from 'Tracking the Black Dog'. PMID- 17087796 TI - The curious incident of methadone in dog's urine. PMID- 17087799 TI - Acculturation and sexual function in Canadian East Asian men. AB - INTRODUCTION: Recent studies have demonstrated the importance of considering acculturation when investigating the sexuality of East Asian women in North America. Moreover, bidimensional assessment of both heritage and mainstream cultural affiliations provides significantly more information about sexual attitudes than simple unidimensional measures, such as length of residency in the Western culture. AIM: The goal of this study was to extend the findings in women to a sample of East Asian men. MAIN OUTCOME MEASURES: Self-report measures of sexual behaviors, sexual responses, and sexual satisfaction. METHODS: Euro Canadian (N = 124) and East Asian (N = 137) male university students privately completed a battery of questionnaires in exchange for course credit. Results. Group comparisons revealed East Asian men to have significantly lower liberal sexual attitudes and experiences, and a significantly lower proportion had engaged in sexual intercourse compared with the Euro-Canadian sample. In addition, the East Asian men had significantly higher Impotence and Avoidance subscale scores on the Golombok Rust Inventory of Sexual Satisfaction, a measure of sexual dysfunction. Focusing on East Asian men alone, mainstream acculturation, but not length of residency in Canada, was significantly related to sexual attitudes, experiences, and responses. CONCLUSIONS: Overall, these data replicate the findings in women and suggest that specific acculturation effects over and above length of residency should be included in the cultural assessment of men's sexual health. PMID- 17087800 TI - A comparison of the efficacy and tolerability of tadalafil 10 mg and 20 mg in Japanese patients with severe erectile dysfunction. AB - INTRODUCTION: Tadalafil is a phosphodiesterase type 5 inhibitor with documented efficacy in the treatment of erectile dysfunction (ED). AIM: To compare the efficacy and tolerability of tadalafil 10 mg and 20 mg in men with severe ED. METHODS: A prespecified subgroup analysis was conducted to compare the efficacy of tadalafil 10 and 20 mg measured by the International Index of Erectile Function (IIEF) erectile function (EF) domain and Sexual Encounter Profile (SEP) among patients with severe ED (EF domain score = 1-10) in a Japanese placebo controlled study (PCT). We also analyzed the efficacy of the two doses in men with severe ED post hoc by pooling data from three tadalafil clinical trials that evaluated these doses using a similar study design (three placebo-controlled trials), and evaluated (post hoc) the presence of organic comorbidities in patients with different levels of response to tadalafil 10 or 20 mg. MAIN OUTCOME MEASURES: Mean change in the IIEF-EF domain and mean per-patient changes in percent "yes" responses to SEP Question 2 (SEP2) and Question 3 (SEP3). RESULTS: Patients with severe ED in the Japanese study experienced numerically greater increases (improvements) when taking tadalafil 20 mg compared with 10 mg in the IIEF-EF domain (14.3 vs. 12.4; P = 0.355), SEP2 (60% vs. 57%; P = 0.781), and SEP3 (61% vs. 49%, P = 0.196). When sufficiently powered, these observations reached statistical significance in the three PCTs: patients with severe ED experienced greater increases when taking tadalafil 20 mg compared with 10 mg in the IIEF-EF domain (13.6 vs. 10.4; P = 0.014) and SEP3 (56% vs. 43%, P = 0.019). Both doses were well tolerated. CONCLUSIONS: Patients with severe ED, and especially those with an organic comorbidity, may derive greater clinical benefits from tadalafil 20 mg compared with 10 mg. PMID- 17087801 TI - Efficacy and patient satisfaction associated with penile prosthesis revision surgery. AB - INTRODUCTION: Complications that arise after placement of a penile prosthesis may result in the need for revision surgery. Few contemporary penile prosthesis series have focused solely on describing the efficacy and patient satisfaction associated with penile prosthesis revision surgery. AIM: To determine the overall success of penile prosthesis revision surgery in providing the patient with a functional implant. Device efficacy and patient satisfaction with penile prosthesis revision surgery will be assessed using psychometrically validated instruments. MAIN OUTCOME MEASURES: The erectile function (EF) and satisfaction domains of the International Index of Erectile Function (IIEF) were used to quantify the overall efficacy and patient satisfaction with revision surgery. METHODS: Data were derived from a prospective database of consecutive patients undergoing penile prosthesis revision surgery performed by a single surgeon (B.R.K.). Eleven patients filled out the IIEF prior to surgery and 26 completed it following surgery. RESULTS: Thirty-nine consecutive patients underwent 55 revision procedures related to a prior penile prosthesis. Thirty-four patients (87%) have a functional implant with a median follow-up of 6.5 months (range 1-42 months). Significant improvements in the overall IIEF, EF domain, and satisfaction domain were seen in those patients studied prior to and following revision surgery. The total IIEF, EF domain, and satisfaction domain of the IIEF for the group of 26 responders were 60.7 +/- 19.3, 26.6 +/- 8.7, and 15.7 +/- 5.6, respectively. Patients with fibrotic corporal bodies scored significantly lower on the EF and satisfaction domains than did any other group. CONCLUSIONS: Penile prosthesis revision surgery is highly successful in providing men with a functional implant. The prostheses function well and patients are satisfied with their devices. Further study of those men with corporal fibrosis is warranted in order to determine the factors needed to improve their overall satisfaction with revision surgery. PMID- 17087802 TI - Erectile dysfunctions in patient-physician communication: optimized strategies for addressing sexual issues and the benefit of using a patient questionnaire. AB - INTRODUCTION: Erectile dysfunctions are prevalent but underdiagnosed and undertreated health problems. Communication barriers between patients and physicians are one of the main reasons for this and responsible for a low report rate of sexual dysfunction. AIM: The main aim of the study was to investigate which phrasing and communication strategies gained the highest acceptance from physicians and their patients and were considered the most effective. MAIN OUTCOME MEASURES: A documentation form on which each consultation was rated by the participating physicians. METHODS: A large group of physicians was asked to hand out a short patient questionnaire to all male patients over 30 years. The physician was instructed to discuss the questionnaire with the patient and to ask him about sexual problems. A total of 1,191 physicians took part in the study that documented a total of 10,622 consultations with an average duration of 15 minutes. RESULTS: The main results were: (i) the patient questionnaire found a high level of acceptance and 54% of discussions of sexual health were prompted by it; (ii) the patients' reaction to physicians addressing sexual health was positive in more than two-thirds of the sample and characterized by openness, willingness to communicate, and relief that their sexual problems had been addressed; (iii) from the physicians' perspective, the most favored communication strategies were a clear signaling of a willingness to talk, and addressing treatment possibilities or signaling that help was available; and (iv) the resulting discussion led to further diagnostic measures in 25% of patients and to further therapeutic measures in 60% of patients. CONCLUSIONS: There are good grounds for concluding that: (i) addressing a patient's sexual health as part of a physician's everyday routine is feasible in terms of duration and content; and (ii) a short patient questionnaire is an excellent aid for patients and physicians for initiating communication on the topic. PMID- 17087803 TI - Sexual outcomes and satisfaction with hysterectomy: influence of patient education. AB - INTRODUCTION: Many women experience improved sexual function after hysterectomy. However, a sizeable minority of women report worsened sexual function after the surgery, and concerns about the effect of surgery on sexual function are common among women planning to undergo hysterectomy. AIM: The present study examined the role of education about the potential sexual consequences of hysterectomy in predicting self-reported outcomes and satisfaction with the procedure. METHODS: We conducted a cross-sectional survey of 204 women who had undergone simple hysterectomy in the preceding 3-12 months. Participants volunteered in response to a Web-based advertisement. MAIN OUTCOME MEASURES: Participants indicated their current sexual function using the Female Sexual Function Index (FSFI), and reported positive and negative sexual outcomes experienced after hysterectomy using a checklist. Participants also completed questionnaire items regarding satisfaction with hysterectomy and education from their physicians about sexual risks and benefits prior to surgery. RESULTS: Current sexual function scores were related to self-reports of positive and negative sexual outcomes following hysterectomy and overall satisfaction with hysterectomy. Education from a physician about possible adverse sexual outcomes was largely unrelated to self reports of having experienced those outcomes. However, education about possible negative sexual outcomes predicted overall satisfaction with hysterectomy when controlling for self-reports of positive and negative sexual outcomes. CONCLUSION: Education about potential negative sexual outcomes after surgery may enhance satisfaction with hysterectomy, independent of whether negative sexual outcomes were experienced. Including a discussion of potential sexual changes after surgery may enhance the benefits of presurgical counseling prior to hysterectomy. PMID- 17087804 TI - Real-life safety and efficacy of vardenafil in the treatment of erectile dysfunction-results from 30,010 U.S. patients. AB - INTRODUCTION: Clinical trials show that vardenafil produces effective and satisfactory first-dose success rates and reliability for erection and intercourse in men with erectile dysfunction (ED). AIM: This study was conducted to evaluate real-life efficacy, safety, and acceptance of vardenafil in men with ED. METHODS: This open-label, prospective study, conducted in 6,740 U.S. centers, included an initial visit and one or two follow-up visits within a 2-month period of the first vardenafil dose. Vardenafil was administered in 5-20 mg doses. MAIN OUTCOME MEASURES: Efficacy variables included first-dose success rates for vaginal penetration, maintenance of erection, and satisfaction based on physician and patient assessments. Safety was assessed by adverse events (AEs). RESULTS: A total of 30,010 men were included in the safety/intent-to-treat (S/ITT) analysis, with 26,043 men in the adjusted S/ITT population. Vardenafil improved erectile function in 78% of men, with 75% rating overall efficacy as "satisfying" or "very satisfying." The overall rates of successful penetration and maintenance with vardenafil following the first dose were 78% and 68%, respectively. For men with mild and moderate ED, first-dose success rates for penetration were 89% and 82%, respectively, and for maintenance, 82% and 71%, respectively. First-dose penetration and maintenance of erection rates were 76% and 66%, respectively, for men with self-reported hypertension, and 70% and 60%, respectively, for men with diabetes mellitus. At study end, 67% of patients preferred to continue using vardenafil. The most frequently reported AEs were headache (4%) and flushing (2%). Vardenafil was well tolerated, with a "satisfied/very satisfied" tolerability rating in 75% of cases as assessed by the physician. CONCLUSIONS: This observational study demonstrated the tolerability and efficacy of vardenafil in men with ED and comorbidities. Vardenafil provided a high rate of first-dose intercourse success and a favorable safety profile in patients with and without comorbid disease. PMID- 17087805 TI - Psychiatric illness presenting with a sexual complaint and management by psychotropic medications: a case report. AB - INTRODUCTION: Sexual medicine healthcare professionals, who do not normally examine men and women with psychiatric disorders, need to be aware that those with psychiatric disorders can and do present with sexual medicine problems. In particular, psychiatric individuals may present with a variety of delusions including those that have sexual content or sexual implications. The rare disorder of reverse delusional misidentification syndrome may be encountered in schizophrenic patients and may be best managed by the combined team effort of a sexual medicine specialist and psychiatrist. AIM: To report a case study that reiterates the assessment and sexual medicine management of a female with sexual dysfunction who believed she was transforming into a male. METHODS: Case report of a woman who attended an outpatient clinic in an academic medical center. RESULTS: A 60-year-old woman with a history of paranoid schizophrenia presented to a gynecologist for ovarian cancer screening. Evaluation revealed complaints that the patient's ovaries were testes that produced sperm and her clitoris was a penis capable of erection and ejaculation. Gynecological examination revealed only atrophic vaginitis. The patient was treated with local minimally absorbed vaginal estrogens and referred for psychological assessment and counseling. Psychotropic medication compliance was encouraged, weekly psychotherapy was continued, and delusional symptoms were minimized. CONCLUSION: Sexual medicine healthcare providers should be prepared to manage sex health concerns of men and women with psychiatric disorders, including delusional misidentification syndrome, in conjunction with a psychiatrist. PMID- 17087806 TI - Subjective sexual response to testosterone replacement therapy based on initial serum levels of total testosterone. AB - INTRODUCTION: Testosterone replacement therapy (TRT) has been shown to be beneficial for men with hypogonadism. However, it is unknown how well hypogonadal men respond to TRT based on the severity of testosterone deficiency. AIM: To determine subjective sexual response rates to TRT based on initial serum testosterone values, with particular interest in men with "low-normal" levels of total testosterone (TT). MAIN OUTCOME MEASURES: Subjective responses to TRT in the domains of erectile dysfunction, libido, orgasm, and morning erections. METHODS: A retrospective study was performed of 211 men with sexual symptoms of hypogonadism who underwent TRT. All men had either low values of TT (<300 ng/dL) or free testosterone (FT) (<1.5 ng/dL). The cohort was divided into three groups based on initial TT levels: Group 1: 0-200 ng/dL (N = 26; 12.3%); Group 2: 201 300 ng/dL (N = 64; 30.3%); Group 3: 301 ng/dL or greater (N = 121; 57.3%). Improvement in erectile function was determined prior to addition of any other treatment (e.g., phosphodiesterase type 5 inhibitors). The mean follow-up was 9 months (range 3-36 months). RESULTS: The mean age was 55.2 years. Testosterone gel was used in approximately two-thirds of each group. Improvement in libido was reported in 61.5%, 96.6%, and 29.8% for Groups 1, 2, and 3, respectively (P < 0.001). Improvement in erectile function was noted in 46.2%, 45.3%, and 73.6% for Groups 1, 2, and 3, respectively (P < 0.001). At time of last follow-up, the percentage of men continuing with TRT was 73.1%, 57.8%, and 58.7% for Groups 1, 2, and 3, respectively (P = nonsignificant). CONCLUSIONS: These preliminary data suggest that men with sexual symptoms of hypogonadism respond well to TRT across a wide range of initial TT values, including men with low-normal TT levels. These men may have low bioavailable levels of testosterone that are not reflected in TT values. PMID- 17087807 TI - Postpartum sexuality concerns among first-time parents from one U.S. academic hospital. AB - INTRODUCTION: Sexual perceptions and activities are influenced by pregnancy and childbirth due to both physical and emotional changes. AIM: Our goal was to illuminate the sexuality issues of concern to first-time parents (mothers and fathers) of healthy, singleton newborns in one U.S. academic hospital. MAIN OUTCOME MEASURES: Number of sexuality concerns and self-reported degree of resolution by months postpartum and gender of parent. METHODS: Anonymous, cross sectional postal survey of two first-time parent cohorts: 4 months postpartum (113 responses/404 mailed = 28.0%) and 12 months postpartum (99 responses/394 mailed = 25.1%). RESULTS: Approximately half of the parents reported questions or concerns on between two and eight of 19 sexuality topics. Only 11% of participating mothers and 17% of participating fathers had one or no concerns (P = 0.27 by gender), and 17% of mothers and 9% of fathers had >13 concerns (P = 0.12 by gender). There were fewer maternal concerns if they did not live with the father (P = 0.01), slightly fewer if the delivery was vaginal vs. cesarean (P = 0.07), and no differences by forceps/vacuum use or episiotomy/vaginal tear. The top concerns at 4 months postpartum were quite similar by gender and included when to resume intercourse, birth control, and the sexual impact of physical recovery from delivery. At 12 months, mothers and fathers both frequently reported a sexuality impact from the mother's body image concerns and desire discrepancy. One year postpartum, there were three sexuality topics with fairly high prevalence (more than one-third parents had this concern) that persisted (no self-reported resolution among at least one-third of parents): child-rearing differences with spouse, greater desire by the man than the woman, and the mother's body image. CONCLUSIONS: New mothers and fathers both have postpartum sexuality concerns/questions, many of which can be addressed by healthcare providers. PMID- 17087808 TI - Patients responding to phosphodiesterase type 5 inhibitor therapy: what do their sexual partners know? AB - INTRODUCTION: Phosphodiesterase type 5 (PDE5) inhibitors are an efficacious therapy in men with erectile dysfunction (ED). There are only a few studies that also focus on the participating couples during PDE5 inhibitor therapy. AIM: To determine to what extent patients personally informed their sexual partners about their ongoing PDE5 inhibitor therapy. MAIN OUTCOME MEASURES: Likelihood of informing the female partner by the patient himself about the use of PDE5 inhibitors. METHODS: A total of 216 men (mean age 62.3 years) with ED were successfully treated with PDE5 inhibitors in three independent centers. After an interval of at least 3 months of successful ED therapy, all patients were asked by questionnaire whether their sexual partners were informed of their PDE5 inhibitor therapy. RESULTS: Eighty-two percent of the patients were exclusively involved in one stable sexual relationship, 9.7% of the men admitted to having changing sexual partners, and 6% did not give any information at all about their sexual partners. Twenty percent of the men had a severe ED (International Index of Erectile Function [IIEF-5] <11). Forty-nine percent showed a moderate ED (IIEF 5 11-16) and 31% suffered a mild ED (IIEF-5 >16). PDE5 inhibitor medication was used 1.2 times/month by men with a severe ED, 2.1 times/month by patients with a moderate ED, and 2.9 times/month by men with a mild ED. Forty-one (93%) of the 44 patients with a severe ED informed their sexual partners that they were taking PDE5 inhibitors. In the patient group with moderate ED, 49 (47%) of 105 patients and only 14 (21%) of 67 of the patients with mild ED shared this information with their partners. CONCLUSION: Less than 40% of the patients suffering a moderate or mild ED using PDE5 inhibitors shared this information with their partners. It seems that patients find ED so disturbing that many patients do not inform their partners of PDE5 inhibitor use. PMID- 17087810 TI - A framework for evidence-based mental health care and policy. AB - OBJECTIVE: Care planning integrates a growing number of disciplines, research fields and analysis techniques. A framework of the main areas of interest with regard to evidence-based health care in mental health is provided here. METHOD: The framework is based on the experience of working with data analysts and health and social decision makers at the PSICOST/RIRAG network, a Spanish research association which includes psychiatrists, health economists and health policy experts, as well as on a review of the literature. RESULTS: Three main areas have been identified and described here: outcomes management, knowledge discovery from data, and decision support systems. Their use in mental health care is reviewed. CONCLUSION: It is important to promote bridging strategies among these new fields in order to enhance communication and information transfer between the different parts involved in mental health decision making: i) clinicians and epidemiologists, ii) data analysts, iii) care policy makers and other end-users. PMID- 17087811 TI - Assessment of service use patterns in out-patients with schizophrenia: a Spanish study. AB - OBJECTIVE: The objective is to describe and characterize patterns of service use by out-patients with schizophrenia in Spain. METHOD: A representative treated prevalence sample of cases with schizophrenia was selected from four Spanish health areas. The evaluation included health service use, clinical severity, functioning and disability. Statistical analysis was based on hierarchical clustering methods. RESULTS: A total of 356 patients were included in the analysis. Five patterns of health service use were defined: heavy out-patient mental health users; mental health and general health service users; heavy hospital service users; nursing service users; low users of mental health services. Patients in each group showed differences in clinical and disability status. Patterns of health service use showed consistency, but also variability, among the geographical areas. CONCLUSION: Development and organization of mental health services should take into account the combinations of services patients most frequently use. PMID- 17087812 TI - Development and usefulness of an instrument for the standard description and comparison of services for disabilities (DESDE). AB - OBJECTIVE: Mental health research has made significant progress in international comparison and instrument development. This study reports the adaptation of the European Service Mapping Schedule (ESMS) to the assessment of services for persons with disabilities. METHOD: Qualitative groups were used to develop the Description and Evaluation of Services for Disabilities in Europe (DESDE). The psychometric analysis of DESDE covered: feasibility, inter-rater reliability, descriptive validity and internal validity. A demonstration study was also carried out. RESULTS: Compared to the original ESMS, a new main branch and several sub-branches were added. We identified 826 services for persons with disabilities, which provided 1284 main types of care. The feasibility and reliability was good for the majority of codes. Only 6% of services were not properly classified. The Boolean factor analysis supported the internal validity of DESDE. CONCLUSION: DESDE is a useful and reliable instrument for the assessment of services for persons with disabilities. PMID- 17087813 TI - Cost-effectiveness of different clinical interventions for reducing the burden of schizophrenia in Spain. AB - OBJECTIVE: To estimate the cost-effectiveness of interventions for reducing the burden of schizophrenia in Spain. METHOD: The study examined the cost effectiveness of seven different types of clinical interventions at the level of Spanish population: i) current situation; ii) older antipsychotics alone; iii) new antipsychotics alone (risperidone); iv) older antipsychotics plus psychosocial treatment; v) new antipsychotics plus psychosocial treatment; vi) older antipsychotics plus case management and psychosocial treatment; vii) new antipsychotics plus case management and psychosocial treatment. RESULTS: Interventions based on the combination of haloperidol with psychosocial treatment or psychosocial treatment plus case management proved to be the most efficient strategies. CONCLUSION: The relatively modest additional cost of concurrent psychosocial treatment has significant health gains, thereby making such a combined strategy for schizophrenia more cost-effective than pharmacology alone. PMID- 17087814 TI - Cost-utility of selective serotonin reuptake inhibitors for depression in primary care in Catalonia. AB - OBJECTIVE: To determine the cost-utility of selective serotonin reuptake inhibitors (SSRIs) for treating depressive disorders prescribed in primary care (PC). METHOD: A total of 301 participants beginning antidepressant treatment with an SSRI were enrolled in a prospective 6-month follow-up naturalistic study. Incremental cost-utility ratios (ICUR) were obtained for several comparisons among different SSRIs. To address uncertainty in the ICUR's sampling distribution, non-parametric bootstrapping was carried out. RESULTS: Taking into account adjusted total costs and incremental quality of life gained, fluoxetine dominated paroxetine and citalopram with 63.4% and 79.3% of the bootstrap replications in the dominance quadrant, respectively. Additionally, fluoxetine was cost-effective over sertraline with 83.4% of the bootstrap replications below the threshold of 33,936 US$/quality-adjusted life year (30,000 euro/QALY). CONCLUSION: Fluoxetine seems to be a better cost-utility SSRI option for treating depressive disorders in PC. PMID- 17087815 TI - Persistence of Bacillus thuringiensis bioinsecticides in the gut of human-flora associated rats. AB - The capability of two bioinsecticide strains of Bacillus thuringiensis (ssp. israelensis and ssp. kurstaki) to germinate and persist in vivo in the gastrointestinal tract of human-flora-associated rats was studied. Rats were dosed either with vegetative cells or spores of the bacteria for 4 consecutive days. In animals fed spores, B. thuringiensis cells were detected in faecal and intestinal samples of all animals, whereas vegetative cells only poorly survived the gastric passage. Heat-treatment of intestinal samples, which kills vegetative cells, revealed that B. thuringiensis spores were capable of germination in the gastrointestinal tract. In one animal fed spores of B. thuringiensis ssp. kurstaki, these bacteria were detected at high density (10(3)-10(4) CFU g(-1) faecal and intestinal samples) even 2 weeks after the last dosage. In the same animal, passage of B. thuringiensis ssp. kurstaki to the spleen was observed; however, no other adverse effects were observed. Denaturing gradient gel electrophoresis of PCR-amplified bacterial 16S rRNA genes in faecal samples revealed no major effect of B. thuringiensis on the composition of the indigenous gut bacteria. Additionally, no cytotoxic effect was detectable in gut samples by Vero cell assay. PMID- 17087816 TI - Lack of in vitro biofilm formation does not attenuate the virulence of Streptococcus gordonii in experimental endocarditis. AB - The ability to induce experimental endocarditis of biofilm-deficient mutants of Streptococcus gordonii was studied in an isogenic background. Strains were inactivated in either comD, fruK or pbp2b genes, which are involved in biofilm formation. These strains were clearly impaired (>75% reduction) in biofilm production in vitro. However, this did not result in a decreased severity of infection in vivo. PMID- 17087817 TI - Breast cancer research: the past and the future. London, UK, 1 November 2006. Abstracts. PMID- 17087818 TI - A second generation radiation hybrid map to aid the assembly of the bovine genome sequence. AB - BACKGROUND: Several approaches can be used to determine the order of loci on chromosomes and hence develop maps of the genome. However, all mapping approaches are prone to errors either arising from technical deficiencies or lack of statistical support to distinguish between alternative orders of loci. The accuracy of the genome maps could be improved, in principle, if information from different sources was combined to produce integrated maps. The publicly available bovine genomic sequence assembly with 6x coverage (Btau_2.0) is based on whole genome shotgun sequence data and limited mapping data however, it is recognised that this assembly is a draft that contains errors. Correcting the sequence assembly requires extensive additional mapping information to improve the reliability of the ordering of sequence scaffolds on chromosomes. The radiation hybrid (RH) map described here has been contributed to the international sequencing project to aid this process. RESULTS: An RH map for the 30 bovine chromosomes is presented. The map was built using the Roslin 3000-rad RH panel (BovGen RH map) and contains 3966 markers including 2473 new loci in addition to 262 amplified fragment-length polymorphisms (AFLP) and 1231 markers previously published with the first generation RH map. Sequences of the mapped loci were aligned with published bovine genome maps to identify inconsistencies. In addition to differences in the order of loci, several cases were observed where the chromosomal assignment of loci differed between maps. All the chromosome maps were aligned with the current 6x bovine assembly (Btau_2.0) and 2898 loci were unambiguously located in the bovine sequence. The order of loci on the RH map for BTA 5, 7, 16, 22, 25 and 29 differed substantially from the assembled bovine sequence. From the 2898 loci unambiguously identified in the bovine sequence assembly, 131 mapped to different chromosomes in the BovGen RH map. CONCLUSION: Alignment of the BovGen RH map with other published RH and genetic maps showed higher consistency in marker order and chromosome assignment than with the current 6x sequence assembly. This suggests that the bovine sequence assembly could be significantly improved by incorporating additional independent mapping information. PMID- 17087819 TI - Epidemiological and clinical features, response to HAART, and survival in HIV infected patients diagnosed at the age of 50 or more. AB - BACKGROUND: Over the last years, the mean age of subjects with HIV infection and AIDS is increasing. Moreover, some epidemiological and clinical differences between younger and older HIV-infected individuals have been observed. However, since introduction of HAART therapy, there are controversial results regarding their response to HAART. The aim of the present study is to evaluate epidemiological and clinical features, response to HAART, and survival in elderly HIV-infected patients with regard to younger HIV-infected patients. METHODS: A prospective cohort study (1998-2003) was performed on patients from Sabadell Hospital, in Northeast of Spain. The cohort includes newly attended HIV-infected patients since January 1, 1998. For the purpose of this analysis, data was censured at December 31, 2003. Taking into account age at time of diagnosis, it was considered 36 HIV-positive people aged 50 years or more (Group 1, G1) and 419 HIV-positive people aged 13-40 years (Group 2, G2). Epidemiological, clinical, biological and therapy data are recorded. Statistical analysis was performed using Chi-squared test and Fisher exact test, Mann-Whitney U test, Kaplan-Meier, Log Rank test, and Two-Way ANOVA from random factors. RESULTS: G1 showed higher proportion of men than G2. The most common risk factors in G1 were heterosexual transmission (P = 0.01) and having sex with men or women (P < 0.001). G1 and G2 show parallel profiles through the time regarding immunological response (P = 0.989) and virological response (P = 0.074). However, older people showed lower CD4 cell counts at first clinic visit (P < 0.001) and, eventually, they did not achieve the same counts as G2. G1 presented faster progression to AIDS (P < 0.001) and shorter survival (P < 0.001). CONCLUSION: Older patients have different epidemiological features. Their immunological and virological responses are good. However, older patients do not achieve the same CD4 cell counts likely due to they have lower counts at first clinic visit. Thus, it is essential physicians know older HIV-infected patients features to consider the possibility of HIV infection in these patients with the aim of treatment would not be delayed. PMID- 17087820 TI - Human TRIM5alpha mediated restriction of different HIV-1 subtypes and Lv2 sensitive and insensitive HIV-2 variants. AB - In order to characterize the antiviral activity of human TRIM5alpha in more detail human derived indicator cell lines over expressing wild type human TRIM5alpha were generated and challenged with HIV-1 and HIV-2 viruses pseudotyped with HIV envelope proteins in comparison to VSV-G pseudotyped particles. HIV envelope protein pseudotyped particles (HIV-1[NL4.3], HIV-1[BaL]) showed a similar restriction to infection (12 fold inhibition) compared to VSV-G pseudotyped viruses after challenging TZM-huTRIM5alpha cells. For HIV-2 a stronger restriction to infection was observed when the homologous envelope protein Env42S was pseudotyped onto these particles compared to VSV-G pseudotyped HIV-2 particles (8.6 fold inhibition versus 3.4 fold inhibition). It has been shown that HIV-2 is restricted by the restriction factor Lv2, acting on capsid like TRIM5alpha. A mutation of amino acid 73 (I73V) of HIV-2 capsid renders this virus Lv2-insensitive. Lv2-insensitive VSV-G pseudotyped HIV-2/I73V particles showed a similar restriction to infection as did HIV-2[VSV-G] particles (4 fold inhibition). HIV-2 envelope protein (Env42S)-pseudotyped HIV-2/I73V particles revealed a 9.3 fold increase in infection in TZM cells but remained restricted in TZM-huTRIM5alpha cells (80.6 fold inhibition) clearly indicating that at least two restriction factors, TRIM5alpha and Lv2, act on incoming HIV-2 particles. Further challenge experiments using primary isolates from different HIV-1 subtypes and from HIV-1 group O showed that wild type human TRIM5alpha restricted infection independent of coreceptor use of the infecting particle but to variable degrees (between 1.2 and 19.6 fold restriction). PMID- 17087821 TI - Evaluation of clustering algorithms for protein-protein interaction networks. AB - BACKGROUND: Protein interactions are crucial components of all cellular processes. Recently, high-throughput methods have been developed to obtain a global description of the interactome (the whole network of protein interactions for a given organism). In 2002, the yeast interactome was estimated to contain up to 80,000 potential interactions. This estimate is based on the integration of data sets obtained by various methods (mass spectrometry, two-hybrid methods, genetic studies). High-throughput methods are known, however, to yield a non negligible rate of false positives, and to miss a fraction of existing interactions. The interactome can be represented as a graph where nodes correspond with proteins and edges with pairwise interactions. In recent years clustering methods have been developed and applied in order to extract relevant modules from such graphs. These algorithms require the specification of parameters that may drastically affect the results. In this paper we present a comparative assessment of four algorithms: Markov Clustering (MCL), Restricted Neighborhood Search Clustering (RNSC), Super Paramagnetic Clustering (SPC), and Molecular Complex Detection (MCODE). RESULTS: A test graph was built on the basis of 220 complexes annotated in the MIPS database. To evaluate the robustness to false positives and false negatives, we derived 41 altered graphs by randomly removing edges from or adding edges to the test graph in various proportions. Each clustering algorithm was applied to these graphs with various parameter settings, and the clusters were compared with the annotated complexes. We analyzed the sensitivity of the algorithms to the parameters and determined their optimal parameter values. We also evaluated their robustness to alterations of the test graph. We then applied the four algorithms to six graphs obtained from high-throughput experiments and compared the resulting clusters with the annotated complexes. CONCLUSION: This analysis shows that MCL is remarkably robust to graph alterations. In the tests of robustness, RNSC is more sensitive to edge deletion but less sensitive to the use of suboptimal parameter values. The other two algorithms are clearly weaker under most conditions. The analysis of high-throughput data supports the superiority of MCL for the extraction of complexes from interaction networks. PMID- 17087822 TI - A simple spreadsheet-based, MIAME-supportive format for microarray data: MAGE TAB. AB - BACKGROUND: Sharing of microarray data within the research community has been greatly facilitated by the development of the disclosure and communication standards MIAME and MAGE-ML by the MGED Society. However, the complexity of the MAGE-ML format has made its use impractical for laboratories lacking dedicated bioinformatics support. RESULTS: We propose a simple tab-delimited, spreadsheet based format, MAGE-TAB, which will become a part of the MAGE microarray data standard and can be used for annotating and communicating microarray data in a MIAME compliant fashion. CONCLUSION: MAGE-TAB will enable laboratories without bioinformatics experience or support to manage, exchange and submit well annotated microarray data in a standard format using a spreadsheet. The MAGE-TAB format is self-contained, and does not require an understanding of MAGE-ML or XML. PMID- 17087823 TI - XML schemas for common bioinformatic data types and their application in workflow systems. AB - BACKGROUND: Today, there is a growing need in bioinformatics to combine available software tools into chains, thus building complex applications from existing single-task tools. To create such workflows, the tools involved have to be able to work with each other's data--therefore, a common set of well-defined data formats is needed. Unfortunately, current bioinformatic tools use a great variety of heterogeneous formats. RESULTS: Acknowledging the need for common formats, the Helmholtz Open BioInformatics Technology network (HOBIT) identified several basic data types used in bioinformatics and developed appropriate format descriptions, formally defined by XML schemas, and incorporated them in a Java library (BioDOM). These schemas currently cover sequence, sequence alignment, RNA secondary structure and RNA secondary structure alignment formats in a form that is independent of any specific program, thus enabling seamless interoperation of different tools. All XML formats are available at http://bioschemas.sourceforge.net, the BioDOM library can be obtained at http://biodom.sourceforge.net. CONCLUSION: The HOBIT XML schemas and the BioDOM library simplify adding XML support to newly created and existing bioinformatic tools, enabling these tools to interoperate seamlessly in workflow scenarios. PMID- 17087824 TI - The complete mitochondrial genome of the sea spider Nymphon gracile (Arthropoda: Pycnogonida). AB - BACKGROUND: Mitochondrial genomes form units of genetic information replicating indepentently from nuclear genomes. Sequence data (most often from protein-coding genes) and other features (gene order, RNA secondary structure) of mitochondrial genomes are often used in phylogenetic studies of metazoan animals from population to phylum level. Pycnogonids are primarily marine arthropods, often considered closely related to chelicerates (spiders, scorpions and allies). However, due to their aberrant morphology and to controversial results from molecular studies, their phylogenetic position is still under debate. RESULTS: This is the first report of a complete mitochondrial genome sequence from a sea spider (Nymphon gracile, class Pycnogonida). Gene order derives from that of other arthropods so that presumably 10 single tRNA gene translocations, a translocation of the mitochondrial control region, and one large inversion affecting protein-coding genes must have happened in the lineage leading to Nymphon gracile. Some of the changes in gene order seem not to be common to all pycnogonids, as those were not found in a partial mitochondrial genome of another species, Endeis spinosa. Four transfer RNAs of Nymphon gracile show derivations from the usual cloverleaf secondary structure (truncation or loss of an arm). Initial phylogenetic analyses using mitochondrial protein-coding gene sequences placed Pycnogonida as sister group to Acari. However, this is in contrast to the majority of all other studies using nuclear genes and/or morphology and was not recovered in a second analysis where two long-branching acarid species were omitted. CONCLUSION: Extensive gene rearrangement characterizes the mitochondrial genome of Nymphon gracile. At least some of the events leading to this derived gene order happened after the split of pycnogonid subtaxa. Nucleotide and amino acid frequencies show strong differences between chelicerate taxa, presumably biasing phylogenetic analyses. Thus the affinities between Pycnogonida and Acari (mites and ticks), as found in phylogenetic analyses using mitochondrial genes, may rather be due to long-branch attraction and independently derived nucleotide composition and amino acid frequency, than to a real sister group relationship. PMID- 17087825 TI - Barriers and motivators to gaining access to smoking cessation services amongst deprived smokers--a qualitative study. AB - BACKGROUND: Smoking is strongly associated with disadvantage and is an important contributor to inequalities in health. Smoking cessation services have been implemented in the UK targeting disadvantaged smokers, but there is little evidence available on how to design services to attract this priority group. METHODS: We conducted focus groups with 39 smokers aged 21-75 from the most socio economically deprived areas of Nottingham UK who had made an unsuccessful attempt to quit within the last year without using smoking cessation services, to identify specific barriers or motivators to gaining access to these services. RESULTS: Barriers to use of existing services related to fear of being judged, fear of failure, a perceived lack of knowledge about existing services, a perception that available interventions--particularly Nicotine Replacement Therapy--are expensive and ineffective, and negative media publicity about bupropion. Participants expressed a preference for a personalized, non judgemental approach combining counselling with affordable, accessible and effective pharmacological therapies; convenient and flexible timing of service delivery, and the possibility of subsidized complementary therapies. CONCLUSION: We conclude that smokers from these deprived areas generally had low awareness of the services available to help them, and misconceptions about their availability and effectiveness. A more personalized approach to promoting services that are non-judgemental, and with free pharmacotherapy and flexible support may encourage more deprived smokers to quit smoking. PMID- 17087826 TI - The 10 kDa domain of human erythrocyte protein 4.1 binds the Plasmodium falciparum EBA-181 protein. AB - BACKGROUND: Erythrocyte invasion by Plasmodium falciparum parasites represents a key mechanism during malaria pathogenesis. Erythrocyte binding antigen-181 (EBA 181) is an important invasion protein, which mediates a unique host cell entry pathway. A novel interaction between EBA-181 and human erythrocyte membrane protein 4.1 (4.1R) was recently demonstrated using phage display technology. In the current study, recombinant proteins were utilized to define and characterize the precise molecular interaction between the two proteins. METHODS: 4.1R structural domains (30, 16, 10 and 22 kDa domain) and the 4.1R binding region in EBA-181 were synthesized in specific Escherichia coli strains as recombinant proteins and purified using magnetic bead technology. Recombinant proteins were subsequently used in blot-overlay and histidine pull-down assays to determine the binding domain in 4.1R. RESULTS: Blot overlay and histidine pull-down experiments revealed specific interaction between the 10 kDa domain of 4.1R and EBA-181. Binding was concentration dependent as well as saturable and was abolished by heat denaturation of 4.1R. CONCLUSION: The interaction of EBA-181 with the highly conserved 10 kDa domain of 4.1R provides new insight into the molecular mechanisms utilized by P. falciparum during erythrocyte entry. The results highlight the potential multifunctional role of malaria invasion proteins, which may contribute to the success of the pathogenic stage of the parasite's life cycle. PMID- 17087827 TI - Lactate levels in severe malarial anaemia are associated with haemozoin containing neutrophils and low levels of IL-12. AB - BACKGROUND: Hyperlactataemia is often associated with a poor outcome in severe malaria in African children. To unravel the complex pathophysiology of this condition the relationship between plasma lactate levels, parasite density, pro- and anti-inflammatory cytokines, and haemozoin-containing leucocytes was studied in children with severe falciparum malarial anaemia. METHODS: Twenty-six children with a primary diagnosis of severe malarial anaemia with any asexual Plasmodium falciparum parasite density and Hb < 5 g/dL were studied and the association of plasma lactate levels and haemozoin-containing leucocytes, parasite density, pro- and anti-inflammatory cytokines was measured. The same associations were measured in non-severe malaria controls (N = 60). RESULTS: Parasite density was associated with lactate levels on admission (r = 0.56, P < 0.005). Moreover, haemozoin containing neutrophils and IL-12 were strongly associated with plasma lactate levels, independently of parasite density (r = 0.60, P = 0.003 and r = -0.46, P = 0.02, respectively). These associations were not found in controls with uncomplicated malarial anaemia. CONCLUSION: These data suggest that blood stage parasites, haemozoin and low levels of IL-12 may be associated with the development of hyperlactataemia in severe malarial anaemia. PMID- 17087828 TI - Assessment of data quality in a multi-centre cross-sectional study of participation and quality of life of children with cerebral palsy. AB - BACKGROUND: SPARCLE is a cross-sectional survey in nine European regions, examining the relationship of the environment of children with cerebral palsy to their participation and quality of life. The objective of this report is to assess data quality, in particular heterogeneity between regions, family and item non-response and potential for bias. METHODS: 1,174 children aged 8-12 years were selected from eight population-based registers of children with cerebral palsy; one further centre recruited 75 children from multiple sources. Families were visited by trained researchers who administered psychometric questionnaires. Logistic regression was used to assess factors related to family non-response and self-completion of questionnaires by children. RESULTS: 431/1,174 (37%) families identified from registers did not respond: 146 (12%) were not traced; of the 1,028 traced families, 250 (24%) declined to participate and 35 (3%) were not approached. Families whose disabled children could walk unaided were more likely to decline to participate. 818 children entered the study of which 500 (61%) self reported their quality of life; children with low IQ, seizures or inability to walk were less likely to self-report. There was substantial heterogeneity between regions in response rates and socio-demographic characteristics of families but not in age or gender of children. Item non-response was 2% for children and ranged from 0.4% to 5% for questionnaires completed by parents. CONCLUSION: While the proportion of untraced families was higher than in similar surveys, the refusal rate was comparable. To reduce bias, all analyses should allow for region, walking ability, age and socio-demographic characteristics. The 75 children in the region without a population based register are unlikely to introduce bias. PMID- 17087829 TI - A rapid and robust method of identifying transformed Arabidopsis thaliana seedlings following floral dip transformation. AB - BACKGROUND: The floral dip method of transformation by immersion of inflorescences in a suspension of Agrobacterium is the method of choice for Arabidopsis transformation. The presence of a marker, usually antibiotic- or herbicide-resistance, allows identification of transformed seedlings from untransformed seedlings. Seedling selection is a lengthy process which does not always lead to easily identifiable transformants. Selection for kanamycin-, phosphinothricin- and hygromycin B-resistance commonly takes 7-10 d and high seedling density and fungal contamination may result in failure to recover transformants. RESULTS: A method for identifying transformed seedlings in as little as 3.25 d has been developed. Arabidopsis T1 seeds obtained after floral dip transformation are plated on 1% agar containing MS medium and kanamycin, phosphinothricin or hygromycin B, as appropriate. After a 2-d stratification period, seeds are subjected to a regime of 4-6 h light, 48 h dark and 24 h light (3.25 d). Kanamycin-resistant and phosphinothricin-resistant seedlings are easily distinguished from non-resistant seedlings by green expanded cotyledons whereas non-resistant seedlings have pale unexpanded cotyledons. Seedlings grown on hygromycin B differ from those grown on kanamycin and phosphinothricin as both resistant and non-resistant seedlings are green. However, hygromycin B-resistant seedlings are easily identified as they have long hypocotyls (0.8-1.0 cm) whereas non-resistant seedlings have short hypocotyls (0.2-0.4 cm). CONCLUSION: The method presented here is an improvement on current selection methods as it allows quicker identification of transformed seedlings: transformed seedlings are easily discernable from non-transformants in as little as 3.25 d in comparison to the 7 10 d required for selection using current protocols. PMID- 17087830 TI - Building health research systems to achieve better health. AB - Health research systems can link knowledge generation with practical concerns to improve health and health equity. Interest in health research, and in how health research systems should best be organised, is moving up the agenda of bodies such as the World Health Organisation. Pioneering health research systems, for example those in Canada and the UK, show that progress is possible. However, radical steps are required to achieve this. Such steps should be based on evidence not anecdotes. Health Research Policy and Systems (HARPS) provides a vehicle for the publication of research, and informed opinion, on a range of topics related to the organisation of health research systems and the enormous benefits that can be achieved. Following the Mexico ministerial summit on health research, WHO has been identifying ways in which it could itself improve the use of research evidence. The results from this activity are soon to be published as a series of articles in HARPS. This editorial provides an account of some of these recent key developments in health research systems but places them in the context of a distinguished tradition of debate about the role of science in society. It also identifies some of the main issues on which 'research on health research' has already been conducted and published, in some cases in HARPS. Finding and retaining adequate financial and human resources to conduct health research is a major problem, especially in low and middle income countries where the need is often greatest. Research ethics and agenda-setting that responds to the demands of the public are issues of growing concern. Innovative and collaborative ways are being found to organise the conduct and utilisation of research so as to inform policy, and improve health and health equity. This is crucial, not least to achieve the health-related Millennium Development Goals. But much more progress is needed. The editorial ends by listing a wide range of topics related to the above priorities on which we hope to feature further articles in HARPS and thus contribute to an informed debate on how best to achieve such progress. PMID- 17087831 TI - Rapid assessment of fine needle aspiration and the final diagnosis--how often and why the diagnoses are changed. AB - BACKGROUND: On-site rapid interpretation (RI) of fine needle aspiration (FNA) has been shown to increase the diagnostic yield of FNA and decrease the need for repeat diagnostic procedures. Because the pathologist interprets only a fraction of the sample and has limited resources available at such times, an occasional RI diagnosis will be changed at the time of the final diagnosis. We investigated how often these changes in diagnoses occur and the possible reasons for the changes. METHODS: All cytology reports from 1/1/02 to 12/31/03 from a single institution were reviewed. Cases with RI with discrepant final diagnoses were noted. The discrepant diagnoses were categorized depending on how they were changed. Possible sources for changed diagnoses were noted. RESULTS: Between 1/1/02 and 12/31/03 there were 1368 RIs of FNAs. Of these 80 (5.8%) had discrepancies between the RIs and final diagnoses. Seventy-eight cases had additional slides and/or cell block at time of final diagnosis. 16 cases had ancillary studies available at final diagnosis. Consultant pathologists were used in 7 cases. Different pathologists interpreted the RI and final diagnosis in 31 cases. CONCLUSION: Although uncommon, discrepancies between RIs and final diagnoses occur 5.8% of the time at our institution. Most commonly, this involves a change of diagnosis from either "non-diagnostic" or "benign" to "malignancy". Although much of this is likely due to the presence of additional material and information at the time of final diagnosis, the number of cases that had different pathologists involved in the RI and final diagnosis suggests that inter-observer variability may also play some role. PMID- 17087832 TI - Pain and psychiatry: a critical analysis and pharmacological review. AB - Pain is one of the most difficult medical problems to diagnose and treat and can be a common symptom of several psychiatric disorders. Pain-related issues are heterogeneous and often underestimated or misinterpreted, with the result that psychiatric interventions, which might have been beneficial from the outset, are often delayed or requested only as a last measure. Several problems arise from the definition, classification and assessment of pain, when documented according to the different scales which are commonly used, since these attempt to cover a multitude of analytical requirements, without really succeeding. An area of constant debate regards the connection between pain and various psychiatric disorders, and the difficulty in the classification of pain disorders within the currently existing framework. The pharmacological treatment of pain is complex and implies a variety of different compounds, from opioids to psychotropic medications like antidepressants and anticonvulsivants. This paper explores the mutual and reciprocal influence between pain and psychiatric disorders reviewing the latest developments in the definition, assessment and treatment of pain, with special emphasis on the impact of pain on psychiatric disorders (and vice versa), and on the use of psychotropic drugs in the treatment of pain syndromes. PMID- 17087833 TI - Development to the blastocyst stage, the oxidative state, and the quality of early developmental stage of porcine embryos cultured in alteration of glucose concentrations in vitro under different oxygen tensions. AB - BACKGROUND: Recent work has shown that glucose may induce cell injury through the action of free radicals generated by autooxidation or through hypoxanthine phosphoribosyltransferase inhibition. The effect of glucose during early in vitro culture (IVC) period of porcine embryos on their developmental competence, contents of reactive oxygen species (ROS) and glutathione (GSH), and the quality of the blastocysts yielded was examined. METHODS: In vitro matured and fertilized porcine oocytes were cultured for the first 2 days (Day 0 = day of fertilization) of IVC in NCSU-37 added with 1.5 to 20 mM glucose (Gluc-1.5 to -20 groups) or pyruvate and lactate (Pyr-Lac group). The embryos in all groups were cultured subsequently until Day 6 in NCSU-37 with 5.5 mM added glucose. The ROS and GSH level were measured at Day 1 and 2. DNA-fragmented nuclei and the total cell numbers in blastocyst were evaluated by TUNEL-staining at Day 6. RESULTS: Under 5% oxygen the blastocyst rates and total cell numbers in the blastocysts in all glucose groups were significantly lower than that in the Pyr-Lac group. Similar result in blastocyst rate was found under 20% oxygen (excluding the Gluc-10 group), but total cell numbers in the blastocysts was similar among the groups. At both oxygen tensions, the H2O2 levels of Day 1 embryos in all glucose groups were significantly higher than that in the Pyr-Lac group, while only the Gluc-1.5 group of Day 2 embryos showed a significantly higher H2O2 level than that in the Pyr-Lac group. The GSH contents of either Day 1 or Day 2 embryos developed under 5% oxygen were similar among the groups. Only the content of Day 2 embryos in 1.5 mM group was significantly lower than the embryos in the Pyr-Lac group under 20% oxygen. Total cell numbers in the blastocysts (except in the Gluc-20 group) were significantly lower in the embryos cultured under 20% oxygen than 5% oxygen. Only the Gluc-20 blastocysts developed under 5% oxygen showed significantly higher DNA fragmentation rate than those of Pyr-Lac blastocysts. CONCLUSION: These results show that a decrease in developmental ability of embryos cultured by use of glucose instead of pyruvate and lactate after the ferilization may be due to the rise in ROS generation in Day 1 embryos. Moreover, results from this study suggest that the concentration of glucose in the medium that can be used by the Day 1-2 embryos is limited to 3.5 mM and exposure to higher glucose concentrations does not improve embryo development. PMID- 17087834 TI - Analysis of the XRCC1 gene as a modifier of the cerebral response in ischemic stroke. AB - BACKGROUND: Although there have been studies of the genetic risk factors in the development of stroke, there have been few investigations of role of genes in the cerebral response to ischemia. The brain responds to ischemia in a series of reactions that ultimately influence the volume of a stroke that, in general, correlates with disability. We hypothesize that polymorphisms in genes encoding proteins involved in these reactions could act as modifiers of this response and impact stroke volume. One of the pathways participating in the cerebral ischemic response involves reactive oxygen species which can cause oxidative damage to nucleic acids. DNA repair mechanisms are in place to protect against such damage and imply a role for DNA repair genes in the response of the brain to ischemia and are potential candidate genes for further investigation. METHODS: We studied two common polymorphisms in the DNA repair gene, XRCC1, C26304T and G28152A, in 134 well characterized patients with non lacunar ischemic strokes. We also performed a case control association study with 113 control patients to assess whether these variants represent risk factors in the development of ischemic stroke. RESULTS: Independent of etiology, the "T" allele of the C26304T polymorphism is significantly associated with larger stroke volumes (T-test analysis, p < 0.044; multivariate regression analysis, beta = 0.23, p < 0.008). In the case control association study, we found that neither of these polymorphisms represented a risk factor for the development of stroke. CONCLUSION: Our study suggests a major gene effect of the "T" allele of the C26304T polymorphism modulating the cerebral response to ischemia in non lacunar ischemic stroke. PMID- 17087835 TI - Diagnostic omission errors in acute paediatric practice: impact of a reminder system on decision-making. AB - BACKGROUND: Diagnostic error is a significant problem in specialities characterised by diagnostic uncertainty such as primary care, emergency medicine and paediatrics. Despite wide-spread availability, computerised aids have not been shown to significantly improve diagnostic decision-making in a real world environment, mainly due to the need for prolonged system consultation. In this study performed in the clinical environment, we used a Web-based diagnostic reminder system that provided rapid advice with free text data entry to examine its impact on clinicians' decisions in an acute paediatric setting during assessments characterised by diagnostic uncertainty. METHODS: Junior doctors working over a 5-month period at four paediatric ambulatory units consulted the Web-based diagnostic aid when they felt the need for diagnostic assistance. Subjects recorded their clinical decisions for patients (differential diagnosis, test-ordering and treatment) before and after system consultation. An expert panel of four paediatric consultants independently suggested clinically significant decisions indicating an appropriate and 'safe' assessment. The primary outcome measure was change in the proportion of 'unsafe' workups by subjects during patient assessment. A more sensitive evaluation of impact was performed using specific validated quality scores. Adverse effects of consultation on decision-making, as well as the additional time spent on system use were examined. RESULTS: Subjects attempted to access the diagnostic aid on 595 occasions during the study period (8.6% of all medical assessments); subjects examined diagnostic advice only in 177 episodes (30%). Senior House Officers at hospitals with greater number of available computer workstations in the clinical area were most likely to consult the system, especially out of working hours. Diagnostic workups construed as 'unsafe' occurred in 47/104 cases (45.2%); this reduced to 32.7% following system consultation (McNemar test, p < 0.001). Subjects' mean 'unsafe' workups per case decreased from 0.49 to 0.32 (p < 0.001). System advice prompted the clinician to consider the 'correct' diagnosis (established at discharge) during initial assessment in 3/104 patients. Median usage time was 1 min 38 sec (IQR 50 sec-3 min 21 sec). Despite a modest increase in the number of diagnostic possibilities entertained by the clinician, no adverse effects were demonstrable on patient management following system use. Numerous technical barriers prevented subjects from accessing the diagnostic aid in the majority of eligible patients in whom they sought diagnostic assistance. CONCLUSION: We have shown that junior doctors used a Web-based diagnostic reminder system during acute paediatric assessments to significantly improve the quality of their diagnostic workup and reduce diagnostic omission errors. These benefits were achieved without any adverse effects on patient management following a quick consultation. PMID- 17087836 TI - Postpartum practices of puerperal women and their influencing factors in three regions of Hubei, China. AB - BACKGROUND: 'Sitting month' is a Chinese tradition for women's postpartum custom. The present study aims to explore the postpartum dietary and health practices of puerperal women and identify their influential factors in three selected regions of Hubei, China. METHODS: A cross-sectional retrospective study was conducted in the selected urban, suburban and rural areas in the province of Hubei from 1 March to 30 May 2003. A total of 2100 women who had given birth to full-term singleton infants in the past two years were selected as the participants. Data regarding postpartum practices and potentially related factors were collected through questionnaire by trained investigators. RESULTS: During the puerperium, 18% of the participants never ate vegetables, 78.8% never ate fruit and 75.7% never drank milk. Behaviour taboos such as no bathing, no hair washing or teeth brushing were still popular among the participants. About half of the women didn't get out of the bed two days after giving birth. The average time they stayed in bed during this period was 18.0 h. One third of them didn't have any outdoor activities in that time periods. The educational background of both women and their spouses, location of their residence, family income, postnatal visit, nutrition and health care educational courses were found to be the influencing factors of women's postpartum practices. CONCLUSION: Traditional postpartum dietary and health behaviours were still popular among women in Hubei. Identifying the factors associated with traditional postpartum practices is critical to develop better targeting health education programs. Updated Information regarding postpartum dietary and health practices should be disseminated to women. PMID- 17087837 TI - DGAT1 and ABCG2 polymorphism in Indian cattle (Bos indicus) and buffalo (Bubalus bubalis) breeds. AB - BACKGROUND: Indian cattle (Bos indicus) and riverine buffalo (Bubalus bubalis) give a poor yield of milk but it has a high fat and protein percentage compared to taurine cattle. The identification of QTLs (Quantitative Trait Loci) on BTA14 and BTA6 and its subsequent fine mapping has led to identification of two non conservative mutations affecting milk production and composition. Our objective was to estimate the frequency of K232A (DGAT1--diacylglycerol-acyltransferase 1) and Y581S (ABCG2--ATP binding cassette sub family G member 2) polymorphisms in diverse cattle and buffalo breeds of India having large variation in terms of milk production. RESULTS: We screened the reported missense mutations in six cattle and five buffalo breeds. The DGAT1K and ABCG2Y alleles were found to be fixed in Indian cattle and buffalo breeds studied. CONCLUSION: This study provides an indirect evidence that all the Indian cattle and buffalo breeds have fixed alleles with respect to DGAT1 and ABCG2 genes reported to be responsible for higher milk fat yield, higher fat and protein percent. PMID- 17087838 TI - Quadricuspid aortic valve not discovered by transthoracic echocardiography. AB - BACKGROUND: Quadricuspid aortic valve is a rare congenital heart defect. Several different anatomical variations of a quadricuspid aortic valve has been described and aortic regurgitation is the predominant valvular dysfunction associated with quadricuspid aortic valve. CASE PRESENTATION: A 68-year-old woman presented with almost a years history of increasing dyspnoea on exertion. The patient have had two previous transthoracic echocardiographic exams in the last six years and they had only documented moderate aortic regurgitation. Transoesophageal echocardiography displayed a rare case of quadricuspid aortic valve with three cusps of equal size and one larger cusp. The malformation was associated with severe aortic regurgitation. CONCLUSION: Liberal use of transoesophageal echocardiography is often warranted if optimal display of valvular morphology is desired. PMID- 17087839 TI - Does adding intravenous fentanyl to caudal block in children enhance the efficacy of multimodal analgesia as reflected in the plasma level of catecholamines? AB - BACKGROUND AND OBJECTIVE: Several studies showed that single analgesic modality management can attenuate perioperative stress, but little is known about the effect of multimodal analgesia on catecholamine responses to surgical trauma in children. METHODS: Fifty children (American Society of Anesthesiologists Grade I or II) were randomly allocated to one of two groups: one received general anaesthesia and a caudal block (control group), and one group was given general anaesthesia, caudal block and intravenous (i.v.) fentanyl 2 microg kg(-1) (fentanyl group). Plasma epinephrine and norepinephrine concentrations were measured three times during the perioperative period: at induction time (T(0)), at the end of surgery (T(1)) and when the children were fully awake in the postanaesthesia care unit (T(2)). RESULTS: There was a significant reduction in the catecholamine levels in the two groups when (T(1)) and (T(2)) were compared with T(0). When plasma epinephrine levels (at T(0), T(1) and T(2)) between the two groups were compared, a statistically significant reduction at T(2) was obtained in the fentanyl group, when compared with the control group. However, plasma norepinephrine levels showed no statistically significant difference between the two groups (at T(0), T(1) and T(2)). CONCLUSION: These findings suggest that the multimodal analgesic approach of adding i.v. low-dose fentanyl to a caudal block may decrease the plasma epinephrine release in children undergoing inguinal herniotomy. PMID- 17087840 TI - Analgesic efficacy of single-dose parecoxib for corneal suturing in children. PMID- 17087841 TI - Relation between bitter taste sensitivity and incidence or intensity of propofol injection pain. AB - BACKGROUND AND OBJECTIVE: In human beings, pain and taste perception are two major sensory inputs. We investigated whether increasing bitter taste sensitivity would increase intensity or incidence of pain associated with propofol, and whether there is a relationship between bitter sensitivity and venepuncture pain. METHODS: One hundred (50 males, 50 females) American Society of Anesthesiologists Grade I adults undergoing elective surgery were included in this study. Determination of the taste thresholds employed a series of propylthiouracil solutions. The filter paper disk method was used to measure the taste threshold. A 20-G intravenous (i.v.) cannula was inserted in the dorsum of the non-dominant hand. Venepuncture pain was assessed by using a numerical rating scale (NRS; 0, no pain and 10, extreme pain). Propofol 10 mL (100 mg) was injected over 30 s. Assessment of pain with i.v. propofol was made using a 4-point scale: 0, no pain; 1, mild pain; 2, moderate pain; 3, severe pain. RESULTS: The NRS score of venepuncture pain was 2.8 +/- 1.5. Sixty patients had pain during propofol injection. There was statistically significant correlation between bitter sensitivity and propofol injection pain, and between bitter sensitivity and venepuncture pain (P < 0.05). CONCLUSIONS: We conclude that increased bitter taste sensitivity correlates with increased intensity or incidence of propofol injection pain and NRS of venepuncture pain. PMID- 17087842 TI - Unexpected tracheal compression detected after immediate extubation failure. PMID- 17087843 TI - Intraoperative anaphylaxis after intravenous atropine. PMID- 17087844 TI - Regional filling characteristics of the lungs in mechanically ventilated patients with acute lung injury. AB - OBJECTIVES: The objective of the study was to determine regional pulmonary filling characteristics in 20 mechanically ventilated patients with acute lung injury. METHODS: Regional filling characteristics were calculated from tracings of regional tidal volumes vs. global tidal volumes measured by electrical impedance tomography (EIT). These plots were fitted to a polynomial function of the second degree. Regional polynomial coefficients of the second degree characterized the curve linearity of the plots. Near-zero values of the polynomial coefficient indicated a homogeneous increase in regional tidal volumes during the whole inspiration. Positive values hinted at initial low regional tidal volume change suggesting lung volume recruitment. Negative values indicated late low regional tidal volume change implying hyperinflation of this lung region. RESULTS: We found a broad heterogeneity of regional lung filling characteristics. The minimal regional polynomial coefficients varied from -2.80 to -0.56 (median -1.16), while the maximal regional polynomial coefficients varied from 0.58 to 3.65 (median 1.41). CONCLUSIONS: Measurements of regional filling characteristics by EIT may be a helpful tool to adjust the respiratory settings during mechanical ventilation to optimize lung recruitment and to avoid overdistension. It applies a non-pressure-related assessment to the mechanics of lung inflation and gives a view of the real problems underlying ventilatory strategies dependent on global characteristics. PMID- 17087845 TI - Effects of storing blood in citrated silicone-coated glass tubes vs. citrated plastic tubes on thromboelastograph variables. PMID- 17087846 TI - Fibre-optic intubation teaching in sedated patients with anticipated difficult intubation. AB - BACKGROUND AND OBJECTIVES: The objective of the study was to assess the safety of training fibre-optic intubation performed under propofol light general anaesthesia in patients with an anticipated difficult intubation. METHODS: Patients with ear, nose and throat cancer having at least two criteria for anticipated difficult intubation and scheduled for fibre-optic intubation were included prospectively. In 26 patients, intubation was performed by an anaesthesia resident (under senior supervision), whereas in 20 patients, it was performed by a senior anaesthesiologist. All patients received propofol light general anaesthesia adjusted to maintain both loss of consciousness and spontaneous ventilation. RESULTS: Of the 46 patients, 45 had successful fibre optic intubation, and one needed a rescue procedure because of hypoxaemia. Residents failed to intubate four patients, who were easily intubated by the senior. Episodic hypoxaemia (SPO2 < 90%) occurred in three patients in each group. No statistically significant difference was found between junior and senior neither on the duration of the procedure (9.3 +/- 4.9 vs. 7.5 +/- 4.0 min) nor on the propofol consumption (197 +/- 130 vs. 193 +/- 103 mg) or the ETCO2 at the end of the procedure (36 +/- 6 vs. 38 +/- 6 mmHg), respectively. CONCLUSION: Teaching fibre-optic tracheal intubation in patients with anticipated difficult intubation and sedated with propofol did not increase morbidity significantly compared with an experienced anaesthesiologist. Fibre-optic intubation under propofol light general anaesthesia could be safely performed by a resident as long as a senior anaesthesiologist is permanently present, spontaneous ventilation is maintained and a rescue oxygenation technique is immediately available. PMID- 17087847 TI - Effect of vital capacity manoeuvres on arterial oxygenation in morbidly obese patients undergoing open bariatric surgery. AB - BACKGROUND: Arterial oxygenation may be compromised in morbidly obese patients undergoing bariatric surgery. The aim of this study was to evaluate the effect of a vital capacity manoeuvre (VCM), followed by ventilation with positive end expiratory pressure (PEEP), on arterial oxygenation in morbidly obese patients undergoing open bariatric surgery. METHODS: Fifty-two morbidly obese patients (body mass index >40 kg m-2) undergoing open bariatric surgery were enrolled in this prospective and randomized study. Anaesthesia and surgical techniques were standardized. Patients were ventilated with a tidal volume of 10 mL kg-1 of ideal body weight, a mixture of oxygen and nitrous oxide (FiO2 = 40%) and respiratory rate was adjusted to maintain end-tidal carbon dioxide at a level of 30-35 mmHg. After abdominal opening, patients in Group 1 had a PEEP of 8 cm H2O applied and patients in Group 2 had a VCM followed by PEEP of 8 cm H2O. This manoeuvre was defined as lung inflation by a positive inspiratory pressure of 40 cm H2O maintained for 15 s. PEEP was maintained until extubation in the two groups. Haemodynamics, ventilatory and arterial oxygenation parameters were measured at the following times: T0 = before application of VCM and/or PEEP, T1 = 5 min after VCM and/or PEEP and T2 = before abdominal closure. RESULTS: Patients in the two groups were comparable regarding patient characteristics, surgical, haemodynamic and ventilatory parameters. In Group 1, arterial oxygen partial pressure (PaO2) and arterial haemoglobin oxygen saturation (SaO2) were significantly increased and alveolar-arterial oxygen pressure gradient (A-aDO2) decreased at T2 when compared with T0 and T1. In Group 2, PaO2 and SaO2 were significantly increased and A-aDO2 decreased at T1 and T2 when compared with T0. Arterial oxygenation parameters at T1 and T2 were significantly improved in Group 2 when compared with Group 1. CONCLUSION: The addition of VCM to PEEP improves intraoperative arterial oxygenation in morbidly obese patients undergoing open bariatric surgery. PMID- 17087848 TI - Postoperative airway obstruction due to Tapia's syndrome after coronary bypass grafting surgery. PMID- 17087849 TI - Are multiple blood transfusions really a cause of acute respiratory distress syndrome? AB - BACKGROUND AND OBJECTIVES: Multiple blood transfusions are considered a common cause of acute respiratory distress syndrome (ARDS). We hypothesized that ARDS is more a consequence of ARDS risk factors (in particular circulatory shock) requiring transfusions than a result of the transfusions themselves. METHODS: This retrospective study included 103 patients admitted during a 10-month period to an 858-bed university hospital who received multiple transfusions (more than six units of packed red blood cells in 24 h). RESULTS: Ten patients developed ARDS; they were more commonly admitted with circulatory shock (36 (38.7%) vs. 8 (80%), P = 0.01), polytrauma (7 (7.5%) vs. 4 (40%), P = 0.01) or thoracic trauma (3 (3.2%) vs. 4 (40%), P = 0.01). The sequential organ-failure assessment (SOFA) score at admission was higher in patients who developed ARDS than in those who did not (9.0 +/- 3.1 vs. 5.6 +/- 3.4, P < 0.005). The total amount of transfusion in the first 24 h was 14.0 +/- 6.8 U in the ARDS patients and 10.6 +/- 7.3 U in the other patients (P = 0.17); the differences remained non-significant in the following days. During the first 24 h, patients who developed ARDS received more fresh frozen plasma than those who did not (21.8 +/- 10.6 U vs. 10.7 +/- 14.7 U, P = 0.02). Patients who developed ARDS had lower PaO2/FiO2 ratios (114 +/- 61 mmHg vs. 276 +/- 108 mmHg, P = 0.01), lower arterial pH (7.27 +/- 0.10 vs. 7.34 +/- 0.11, P = 0.06) and higher minute volume (10.6 +/- 2.8 L min(-1) vs. 7.9 +/- 1.8 L min(-1), P = 0.03) than patients without ARDS. Multivariable analysis retained thoracic trauma and hypoxaemia during the first 24 h (but not multiple transfusions) as independent risk factors for ARDS. CONCLUSIONS: In this retrospective study, the development of ARDS in massively transfused patients was less related to poly-transfusion than to other factors related to circulatory shock, polytrauma or thoracic trauma. Thoracic trauma and a low PaO2 during the first 24 h were identified as independent risk factors for ARDS. PMID- 17087850 TI - Psychomotor dysfunction after remifentanil/propofol anaesthesia. AB - BACKGROUND AND OBJECTIVES: Early recovery after anaesthesia is gaining importance in fast track management. The aim of this study was to quantify psychomotor recovery within the first 24 h after propofol/remifentanil anaesthesia using the Short Performance Test (Syndrom Kurztest (SKT)), consisting of nine subtests. The hypothesis was that psychomotor performance remains reduced 24 h after anaesthesia. METHODS: Thirty-seven patients scheduled for elective surgery took part in the study. The SKT was performed on the day before general anaesthesia (T0), 10, 30, 90 min and 24 h after extubation (T1). Parallel versions were used to minimize learning effects. Anaesthesia was introduced and maintained with remifentanil/propofol as a target controlled infusion. Propofol plasma concentration was measured 10 and 90 min after extubation. Perioperative pain management included novaminsulfon and piritramide. RESULTS: Up till 90 min after surgery and anaesthesia, psychomotor performances were significantly reduced as the lower test results in all SKT subtests indicated (P < or = 0.007 vs. baseline T0). In the three memory subtests (ST 2, ST 8 and ST 9), psychomotor performance was still reduced on the first postoperative day (P < or = 0.005; T1 vs. T0). There was no correlation between propofol plasma concentration and the psychometric test results. CONCLUSIONS: Propofol/remifentanil-based target controlled general anaesthesia for surgery is associated with a reduced psychomotor function up to the first postoperative day. Further studies are needed to confirm the usefulness of the SKT in the perioperative period and to clarify which components in the perioperative period are responsible for a lower performance in the SKT. PMID- 17087851 TI - The interdependence of sleep apnea, excess body weight, and hypertension: A triangular relationship. PMID- 17087852 TI - Rimonabant--a new weapon in the war on obesity. PMID- 17087853 TI - Benefits of renin-angiotensin system blockade in advanced renal insufficiency. PMID- 17087854 TI - Stress signaling in the heart by AMP-activated protein kinase. AB - The stress-signaling protein, adenosine monophosphate-activated protein kinase (AMPK), regulates a variety of pathways in cells that 1) increase the provision and utilization of energy-providing substrates such as glucose and fatty acids, 2) inhibit energy-requiring pathways such as cholesterol biosynthesis and protein synthesis, and 3) increase the transcription of genes involved in energy metabolism and mitochondrial biogenesis. In the heart, AMPK therefore becomes very important in protecting against ischemia-reperfusion injury and regulating substrate metabolism in the face of changes in workload. This review summarizes the regulation of AMPK activity in the heart and discusses the effects of AMPK activation. PMID- 17087855 TI - Obesity and insulin resistance: effects on cardiac structure, function, and substrate metabolism. AB - It is widely recognized that obesity and insulin resistance can contribute to an increased risk of coronary disease, but it has also become increasingly apparent that they may contribute directly to cardiac dysfunction even in the absence of significant coronary disease. Recently, obesity, which is frequently accompanied by insulin resistance, has been independently related to clinically diagnosed heart failure. Thus, there is renewed interest in the pathophysiology of myocardial disease related to obesity and insulin resistance, as well as in the specific cellular mechanisms by which obesity may cause detrimental cardiac structural and functional changes. Alterations in hemodynamics, plasma volume, neurohormonal status, and myocardial substrate metabolism all appear to contribute to these changes. Improving our understanding of cardiac dysfunction related to obesity and insulin resistance may provide clues for new strategies to prevent and treat this alarmingly prevalent condition. PMID- 17087856 TI - On the hypothesis that the failing heart is energy starved: lessons learned from the metabolism of ATP and creatine. AB - Adenosine triphosphate (ATP) and phosphocreatine fall in the failing heart. New insights into the control of ATP synthesis, supply, and utilization, and how this changes in the failing heart, have emerged. In this article, we address four questions: What are the mechanisms explaining loss of ATP and creatine from the failing heart? What are the consequences of these changes? Can metabolism be manipulated to restore a normal ATP supply? Does increasing energy supply have physiologic consequences (ie, does it lead to improved contractile performance)? In part 1 we focus on ATP, in part 2 on creatine, and in part 3 on the relationship between creatine and purine metabolism and purine nucleotide signaling. PMID- 17087857 TI - Deranged energy substrate metabolism in the failing heart. AB - Control of energy metabolism in the heart is closely linked to cardiac performance. Dysregulation of energy-generating pathways occurs in many forms of heart disease, including heart failure. Uncertainty exists as to whether these alterations in the way adenosine triphosphate (ATP) is produced serve to protect the heart from excessive oxygen demands or have untoward long-term consequences. Regulation of fatty acid beta-oxidation (FAO), the principal source of ATP in the healthy heart, occurs at multiple levels, including a strong gene transcriptional component. In the heart, members of the peroxisome proliferator-activated receptor (PPAR) family of transcription factors are the primary regulators of FAO gene expression. PPARs are ligand activated by endogenous lipids and synthetic small molecules, thus providing attractive targets for pharmaceutical intervention. This article discusses controversies surrounding our understanding of cardiac energy metabolism in heart failure and the role that PPAR family members may play, either as contributors to or as potential adjunctive therapy for cardiac disease. PMID- 17087858 TI - Advanced glycation end products and vascular structure and function. AB - Diabetes mellitus has now reached epidemic proportions in the Western world. The associated microvascular and macrovascular complications are a result of severe metabolic derangement, which leads to chronic tissue injury. Although there are a number of proposed pathophysiologic mechanisms for the vascular complications associated with diabetes, this review focuses predominantly on the role of advanced glycation end products (AGEs) in the pathogenesis of diabetes-associated atherosclerosis. The potential role of AGEs in enhancing arterial stiffness, an entity occurring with a greater prevalence in populations known to have higher than-normal AGE levels, is also examined. Pharmacologic interventions aimed at reducing the level of these chemical compounds or interrupting their action provide hope for the future treatment of both atherosclerotic vascular disease and systolic hypertension, particularly in the setting of diabetes. PMID- 17087859 TI - Management of hypertension and dyslipidemia. AB - Cardiovascular disease, the leading cause of death in the United States and other developed societies, can be managed with intensive risk factor modification, including treatment of hypertension and dyslipidemia. Evidence for reduction of cardiovascular morbidity and mortality is summarized in evidence-based guidelines, primarily for hypertension in the Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation and Treatment of High Blood Pressure and for dyslipidemia in the Third Report of the National Cholesterol Education Program Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults. Many medications exist for treatment of hypertension and a limited number for dyslipidemia; the cornerstone of therapy for both remains lifestyle modification, including dietary interventions. Despite guidelines, control rates of concomitant hypertension and dyslipidemia remain low. Patient adherence may affect achievement of recommended goals of therapy for dyslipidemia and hypertension. Effective education and communication may improve overall achievement of treatment goals. PMID- 17087860 TI - Pathophysiology and medical management of systemic hypertension in preeclampsia. AB - Hypertension that complicates preeclampsia in pregnancy is a disorder that requires special consideration in both prevention and pharmacologic treatment. In recent years, few advances have been made regarding the pathophysiology and prevention of preeclampsia; however, there have been some promising results from studies on possible modes of screening women for preeclampsia before clinical signs and symptoms are apparent. The recommendations for first-line drug therapy for the hypertensive complications of preeclampsia have changed little, primarily because first-line medications have had the advantage of extensive research experience. Recent clinical trials have demonstrated the efficacy and safety of various second-line drugs for the hypertensive complications of preeclampsia; whether these therapies can eventually replace the standard recommended first line medications will require more extensive long-term investigation. PMID- 17087861 TI - Role of community programs in controlling blood pressure. AB - Despite more than 30 years of intense activity to improve control--and more recently prevention--high blood pressure continues to be a major public health problem. Evidence-based reviews have identified best practices and quality improvement strategies to address prevention and control. Since the 1970s, community-based programs have been instrumental in raising awareness, increasing knowledge, and promoting changes in health behavior to improve blood pressure control. Most of these programs have emphasized the use of partnerships and involvement of community residents in conducting screening and referral activities, implementing clinical practice guidelines, and increasing healthy eating and physical activity. Many also have used health care team approaches, including the use of trained community health workers to deliver targeted, culturally sensitive heart health education, particularly related to the prevention of cardiovascular disease risk factors in general and high blood pressure in particular. Increased focus on implementation of evidence-based lifestyle and clinical management strategies coupled with community-based approaches may help increase blood pressure control rates within communities. PMID- 17087862 TI - Renovascular hypertension update. AB - Hypertension detected in patients with renovascular disease poses a major clinical challenge. The rapid expansion of noninvasive imaging, effective antihypertensive drug therapy, and endovascular interventional procedures combine to make optimal management a moving target. Renal arterial disease accelerates the development of hypertension associated with activation of multiple pressor systems and accelerated target organ injury. Younger individuals with fibromuscular lesions often respond well to renal revascularization with minor associated risks. Care must be taken in cases of complex vascular anomalies, such as renal artery aneurysms. Atherosclerotic renal artery stenosis is detected more commonly than ever before and affects more than 85% of patients referred for revascularization. Most are older patients with long-standing hypertension, diabetes, and pre-existing complications of vascular disease. The benefits of extensive workup and intervention in this group of patients are controversial. Antihypertensive drug therapy is most effectively achieved with drugs that block the renin-angiotensin system, but most require multiple agents. Selection of patients for renal revascularization in this group is far more controversial than with fibromuscular disease. Several small trials failed to identify major benefits with renal artery angioplasty as compared to closely monitored drug therapy, although crossover rates from medical to interventional arms were high. The Cardiovascular Outcomes in Renal Atherosclerotic Lesions (CORAL) seeks to randomly assign subjects with proven, high-grade renal artery lesions to optimal medical management with and without stenting. This important trial employs distal embolic protection to prevent deterioration of renal function. Understanding the optimal role for renal revascularization depends heavily upon the successful conduct of such trials. PMID- 17087863 TI - How do we know that the pain in fibromyalgia is "real"? AB - Fibromyalgia is a common idiopathic pain condition often resulting in increased morbidity and disability in patients. The lack of peripheral abnormalities in this disease has led clinicians and researchers alike to question if this syndrome represents a valid entity. Recent genetic findings suggest that specific gene mutations may predispose individuals to develop fibromyalgia. In addition, neurobiological studies indicate that fibromyalgia patients have abnormalities within central brain structures that normally encode pain sensations in healthy pain-free controls. Future studies that focus on central neurobiological and/or genetic influences in fibromyalgia may bring insight into mechanisms of this problematic disease and ultimately result in improved treatments. PMID- 17087864 TI - Temporomandibular joint pain and dysfunction. AB - Pain caused by temporomandibular disorders originates from either muscular or articular conditions, or both. Distinguishing the precise source of the pain is a significant diagnostic challenge to clinicians, and effective management hinges on establishing a correct diagnosis. This paper examines terminology and regional anatomy as it pertains to functional and dysfunctional states of the temporomandibular joint and muscles of mastication. A review of the pathophysiology of the most common disorders is provided. Trends in evaluation, diagnosis, treatment, and research are presented. PMID- 17087865 TI - Giant cell arteritis. AB - Giant cell, or temporal, arteritis is a vasculitis of the medium and large arteries that preferentially involves vessels originating from the arch of the aorta. Classically, this disease manifests in an older individual with new-onset persistent headache, an abnormal temporal artery on examination, and increased serum inflammatory markers. The level of clinical suspicion for giant cell arteritis should be based upon patient age, clinical symptoms, and laboratory evaluation. However, the diagnostic gold standard is achieved by histologic confirmation by temporal artery biopsy. Prompt treatment with corticosteroids is essential in order to minimize the frequency of permanent sequelae such as visual loss and stroke. PMID- 17087866 TI - Chronic low back pain: progress in therapy. AB - Low back pain is a common complaint in the primary care setting. Although most patients with acute low back pain will improve with conservative treatment, back pain often recurs, and a subset of patients will progress to chronic and sometimes disabling symptoms. A variety of treatments have been used for chronic low back pain; the best evidence supports intensive rehabilitation programs that include tailored exercise therapy in conjunction with cognitive behavioral interventions and education. The benefit of medications, including nonsteroidal anti-inflammatory drugs, analgesics, and possibly antidepressants or muscle relaxants, must be weighed against potential adverse effects. There is insufficient evidence regarding the effectiveness of many of the available therapies for chronic low back pain, and well-designed, randomized controlled trials are needed to clarify the role of these treatments. PMID- 17087867 TI - Legal aspects of chronic opioid therapy. AB - Although many Americans suffer from undertreated pain, the regulatory and legal environment for the use of opioids in pain relief is currently in a state of flux. The federal government's efforts to curb drug abuse have complicated the use of opioids for pain relief. Recent actions by the US Drug Enforcement Agency have added to an atmosphere of mistrust and confusion and have increased physician concerns about increased scrutiny and legal, regulatory, or administrative sanctions. Despite a disturbing shift in regulatory authority over opioid analgesics away from health agencies and toward law enforcement agencies, recent state policies and guidelines from national medical organizations are playing an important role in promoting the use of opioids for pain treatment and helping to reduce practitioners' concerns over regulatory oversight. Current and future trends concerning the legal and regulatory aspects of chronic opioid treatment are discussed in this article. PMID- 17087868 TI - Chronic low back pain. AB - Low back pain (LBP) is one of the most prevalent medical problems in society today. In addition to the profound effect LBP can have on patients, it has an exceedingly high societal cost. Although most acute episodes of back pain will ultimately resolve, this condition will become chronic for many. Those with chronic LBP are a challenge to treat. Nonetheless, with a better understanding of the underlying pathophysiology and treatment options, our management of these patients is gradually improving. Conservative therapy remains the mainstay treatment of chronic LBP. If this has failed, surgical options may be considered in the carefully selected patient. Fusion is the most established treatment option for this condition. Disc arthroplasty is being increasingly considered. Class I studies critically evaluating established and evolving technologies continue to help shape our understanding of the surgical options for this condition. PMID- 17087869 TI - Tension-type headache and psychiatric comorbidity. AB - Much of the contemporary literature on headache disorders focuses on migraine headaches, despite the fact that tension-type headache (TTH) is highly prevalent and can be as debilitating as migraines. This article reviews the current literature on prevalence rates of psychiatric disorders in TTH populations, psychologic factors associated with TTH, and psychiatric disorders and their relationships with treatment outcomes in TTH. Key conclusions of this review include 1) prevalence rates of TTH vary across clinical and population-based samples; 2) greater TTH chronicity is associated with increased affective distress; 3) Axis II personality disorders may play an important role in TTH prevalence rates and psychologic functioning but have been understudied to date; and 4) maladaptive coping is common in persons with TTH. PMID- 17087870 TI - Tension-type headache in the elderly. AB - Tension-type headache is the most common primary headache disorder seen in adults. Although the prevalence peaks in the fourth and fifth decades of life, significant fractions of the elderly continue to experience either episodic or chronic tension-type headaches. Many secondary headache disorders may present with headaches symptomatically compatible with the diagnosis of tension-type headache. Because numerous secondary headache disorders are more likely to be seen in older adults, the diagnosis of tension-type headache requires vigilance for and exclusion of organic disease. Once the diagnosis of tension-type headache is made, numerous nonpharmacologic and pharmacologic management options are available to reduce the frequency and severity of episodes. Special dosing considerations must be considered in the elderly. Advancing age is a positive prognostic factor in the remission of episodic and chronic tension-type headaches. PMID- 17087871 TI - Tension-type headache: why does this condition have to fight for its recognition? AB - Tension-type headache is the most prevalent type of primary headache but is frequently forgotten by the patient and doctor. This article summarizes the present knowledge about tension-type headache and discusses some of the problematic aspects. Most patients in specialized headache clinics suffer from several different primary and secondary headaches at the same time and deserve a careful characterization before a rational therapy can be initiated. In particular, it is of utmost importance to increase focus on clinical and basic research in order to develop effective treatment strategies for the most neglected and most costly type of headache. PMID- 17087872 TI - Burden of tension-type headache. AB - Epidemiology of headache has been the subject of increased attention recently. It appears that the overall human and financial cost of headache is considerable. Moreover, when the comorbidity and indirect implications of headache are taken into account, the result can be staggering. Most of the literature has concentrated on migraine rather than other headache types such as tension-type headache. This article gathers emerging data that give an estimate of some of the aspects of the burden imparted by tension-type headache on society. It also briefly considers some of the factors that could positively influence this challenge of modern medicine. PMID- 17087873 TI - At the crossroads between tension-type headache and fibromyalgia. AB - Fibromyalgia syndrome and tension-type headache have multiple clinical features in common, and pathogenic mechanisms partly overlap. Significant differences need to be recognized as well. Studying the correlations of these often comorbid conditions represents a unique opportunity to gain insight into their pathophysiology and that of other chronic pain syndromes, to increase the accuracy of their diagnosis, and to improve the therapeutic armamentarium. PMID- 17087874 TI - Tension-type headache in the young. AB - Headache is common in childhood; recurrent headaches include both migraine and tension-type headache (TTH). Although much of the attention recently has been focused on migraine, TTH can also be problematic for children and needs further study. Standardized criteria have been developed and used for epidemiology and treatment studies, but they are yet to be validated for children. The pathophysiology, selective treatment, impact, and outcome of TTH in children need to be studied further. PMID- 17087875 TI - Metallothionein I + II expression and roles during neuropathology in the CNS. PMID- 17087876 TI - Aspects of the molecular and functional genetics in T1DM: a study of selected candidate genes. PMID- 17087877 TI - Studies on serum YKL-40 as a biomarker in diseases with inflammation, tissue remodelling, fibroses and cancer. PMID- 17087878 TI - Prevalent pain and pain level among torture survivors: a follow-up study. AB - AIM: To estimate change over nine months and over two years, as concerns the prevalence and level of pain in the head, back and feet, among previously tortured refugees settled in Denmark, and to compare associations between torture methods and the prevalence of pain at baseline and at follow-up. METHODS: Sixty nine refugees previously exposed to torture in their home country were interviewed at a Danish rehabilitation clinic. Fifty-four accepted to be re interviewed after nine months, and 47 were interviewed again 14 months later. Interviews focussed on the history of exposure to physical and mental torture and on pain in the head, back and feet at baseline and at follow-up. RESULTS: The mean cumulative duration of imprisonment was 1.7 years, and on the average more than 10 years elapsed between torture and examination. The most frequent physical torture method reported was beating (97%), whereas the main mental torture method was threats of death (97%). The prevalence of pain reported at the follow-up interviews did not differ significantly from that reported at baseline (pain in the head, 81% at baseline and 77% at 23-month follow-up; back, 78% and 81%; feet, 59% and 70%). The same pattern was found when examining the level of pain as indicated by Visual Analogue Scales. Pain in the feet at follow-up was associated with previous exposure to beating against the feet. Pain in the back at baseline and pain in the head at follow-up were associated with suffocation. CONCLUSION: More than ten years after the torture took place, survivors of torture continue to suffer from pain associated with the type of torture they had been subjected to. This presents a considerable challenge to future evidence-based development of effective treatment programmes. PMID- 17087880 TI - Conservative treatment of rotator cuff injuries. AB - Across all ages and activity levels, rotator cuff injuries are one of the most common causes of shoulder pain. The anatomy and biomechanics of the shoulder guide the history and physical exam toward the appropriate treatment of rotator cuff injuries. Rotator cuff tears are rare under the age of 40 unless accompanied by acute trauma. Throwing athletes are prone to rotator cuff injury from various causes of impingement (subacromial, internal, or secondary) and flexibility deficits, strength deficits, or both along the kinetic chain. Most rotator cuff injuries may be treated conservatively by using regimens of nonsteroidal anti inflammatory drugs (NSAIDs), corticosteroid injections, and functional rehabilitation therapy. Injury prevention programs are essential for the long term care of patients with rotator cuff disease, for primary prevention, and for prevention of recurrent injuries, unless a traumatically torn rotator cuff is present. Surgical management is reserved for refractory cases that have exhausted conservative measures. PMID- 17087881 TI - Imaging in evaluating the rotator cuff. AB - Multiple modalities are available for imaging the shoulder, including radiography, arthrography, computed tomography (CT), CT arthrography, ultrasound (US), magnetic resonance (MR) imaging, and MR arthrography. Each of these modalities has advantages and disadvantages in the evaluation of shoulder structures. This article discusses imaging of the rotator cuff of the shoulder with a primary focus on evaluation by MR imaging and MR arthrography. PMID- 17087882 TI - Rotator cuff injuries in the contact athlete. AB - Rotator cuff injuries in contact athletes are typically a result of a different mechanism than that seen with older patients or overhead athletes. This unique mechanism along with the extreme demands of these athletes presents special challenges to the surgeon. Special consideration should be given to in-season rehabilitation to allow the athlete to continue to compete if possible. When this is not possible, or for the out-of-season athlete with a rotator cuff injury requiring surgery, special consideration must be given to obtaining the strongest repair possible. The goal in all aspects of treatment is returning the athlete to full participation safely while minimizing recurrent injury or disability during the recovery process. PMID- 17087883 TI - Mini-open rotator cuff repair. AB - Open shoulder procedures require a deltoid release for proper exposure. Arthroscopic techniques have progressed so that minimally invasive techniques give similar outcomes as more formal open procedures with less risk of morbidity. Arthroscopically assisted open rotator cuff repair offers advantages over open procedures with some diagnostic and decompression performed with the arthroscope. The mini-open technique has more aspects of a cuff repair performed through the arthroscope leaving a few steps to be done open. The modern use of arthroscopic techniques for minimally invasive rotator cuff surgery coupled with advances in rehabilitation is discussed. PMID- 17087884 TI - Technique for arthroscopic rotator cuff repair. AB - Modern arthroscopic tools and techniques have allowed surgeons to operate on a wide variety of injuries using procedures less invasive than traditional open methods. For shoulder surgery in general, and rotator cuff repairs specifically, methods now yield a similar footprint as open procedures with several advantages, including reduced tissue trauma, postoperative pain, swelling, and concern about the deltoid attachment, which should lead to good outcomes. PMID- 17087885 TI - Demographics of high-energy mechanisms of injury in the Kids Inpatient Database. AB - The purpose of this study was to review the relationship of patient demographics to mechanism of injury (MOI). The 2000 Kids Inpatient Database (KID) was used. Logistic regression was used to evaluate the relationship between each MOI relative to other MOIs for each of five identified predictors (age, gender, race, socioeconomic status, geographic region). The KID had 87,795 children with a MOI coded and complete data for all predictors. For motor vehicle accidents, 16- to 20-year-olds were up to 3.72 times more likely to be involved than any other age group, and males were 40% less likely compared with females. For firearm hospitalizations, 16- to 20-year-old black males have significantly higher risk compared with all other identified groups. PMID- 17087886 TI - A biomechanical evaluation of an interspinous device (Coflex) used to stabilize the lumbar spine. AB - A biomechanical study of an interspinous stabilization spinal implant (Coflex) was carried out using eight human lumbar L4/L5 motion segments. Each motion segment was tested in compression, then flexion/extension, then lateral bending, and then axial rotation at five conditions: 1) intact; 2) partial destabilization (by cutting the supraspinous and interspinous ligaments, the ligamentum flavum, the facet capsules, and 50% of the inferior bony facet bilaterally); 3) stabilization with the Coflex device; 4) complete destabilization with total laminectomy; and 5) stabilization with pedicle screws and rods. The most important result is that the motion segment after destabilization and insertion of the Coflex device does not allow significantly more or less motion than the intact specimen in either flexion/extension or axial rotation. Thus the Coflex offers nonrigid fixation and can return a partially destabilized specimen back to the intact condition in terms of motion in flexion/extension and axial rotation. PMID- 17087887 TI - Current concepts in locked plating. AB - Approaches to internal fixation have become more biologic. Greater emphasis is placed on vascularity and soft tissue integrity. Locked plates, analogous to rigid internal fixators, can provide relative stability favorable to secondary fracture healing. If applied appropriately, they can avoid soft tissue compromise. The key to this new generation of plates is the locking mechanism of the screw to the plate, which provides angular stability and avoids compression of the plate to the periosteum. Favorable biomechanical and clinical results continue to expand the number of appropriate indications for use of locked plating devices, although exact indications for their use have yet to be precisely defined. PMID- 17087888 TI - Management of metallic mercury injection in the hand. AB - This case report discusses a patient who injected 20 mL of metallic mercury subcutaneously in his left hand during an attempted suicide. The blood mercury level was 118 microg/L and the urinary mercury level was 43 microg/L, which confirmed the diagnosis of metallic mercury poisoning. A good result was obtained in this patient and the local and systemic toxicity and its management are discussed. A chelation therapy with dimercaprol and early surgical excision of injected material are recommended because this treatment effectively lowers mercury blood levels and controls the local inflammatory reaction. Intraoperative fluoroscopy is useful to confirm the extent of removal. PMID- 17087889 TI - Ring removal from a swollen finger: a refined technique. AB - A ring or circular object (wide steel band/nut) impaled on a swollen finger can be most difficult to remove. Likewise, wedding rings in the preoperative setting do not necessarily need to be cut off. This not previously described method uses a Penrose drain to wrap out the finger, an Esmarch or Ace elastic bandage to exsanguinate the extremity, and a constantly inflated surgical tourniquet or blood pressure cuff to keep out any influx of blood while the swollen finger is being decompressed repeatedly with a reapplication of the Penrose drain. The equipment for removal is readily available in all medical offices or emergency departments. This article presents a solution to a common problem. PMID- 17087890 TI - Re: Adolescent idiopathic scoliosis: modern management guidelines, Shindle, M.K., Khanna, A.J., Bhatnagar, R., Sponseller, P.D., JSOA 15(1):43-52, 2006. PMID- 17087891 TI - Effect of postpartum intrauterine treatment with 2% polyvinyl-pyrrolidone-iodine solution on reproductive efficiency in cows. AB - The effect of a routine postpartum intrauterine infusion of 50 to 100 ml 2 % polyvinylpyrrolidone (PVP) -iodine solution on the reproductive performance of dairy cows was investigated in field trials. At a regular herd visit, alternate cows (n = 531) at around one month after calving (maximum 50 and minimum 20) were treated in utero with PVP-iodine. Additional cows (n = 474) were left untreated or were infused in utero with physiological saline and served as controls. The first service conception rate, overall conception rate within 180 d after calving, and average interval in days from parturition to conception were 48.5 and 74.5 %, 98 +/- 36 d in the treated group and 54.9 and 75.9 %, 90 +/- 34 d in the controls. Thus, the routine postpartum treatment with 2 % PVP-iodine solution was not effective in improving the reproductive efficiency of cows. The results were nearly the same in all groups, across herd, year of trial, parity, interval from parturition to treatment and dose of PVP-iodine solution. Besides, for cows with endometritis, an intrauterine infusion of PVP-iodine solution was not only ineffective but it was detrimental to fertility. PMID- 17087892 TI - Relationship of uterine infections and folliculogenesis in dairy cows during early puerperium. AB - Postpartum uterine infections have been associated with reduced fertility in dairy cows; however, the mechanism by which uterine infections limit reproductive function has not been clearly determined. The purpose of this study was to elucidate the relationship between uterine infections in the early puerperium and the onset of folliculogenesis in dairy cows. The pattern and intensity of uterine infections and follicular dynamics were studied in cows that shed fetal membranes (n = 18), and in those that retained fetal membranes after parturition (n = 18). Endometrial swabs collected aseptically from each animal on Days 4, 8 and 12 after parturition were cultured. Ultrasound scanning of the ovaries was carried out on Days 4, B and 12 using a B mode, real-time, linear array ultrasound scanner. The total number of follicles was recorded, and the follicles were classified according to size as small (2 to 4 mm) or medium (5 to 7 mm). The severity of infection was higher (P<0.05) in retained placenta cows on Days 4, 8 and 12 compared to nonretained placenta cows. The total number of follicles was larger (P<0.05) in nonretained placenta cows on Days 4, 8 and 12 than in retained placenta cows. The distribution of different sizes of follicles on all days of observations was similar in both groups of cows (P>0.05). These data support the hypothesis that uterine infection may delay the initiation of folliculogenesis and suppress the rate of follicular growth in dairy cows in the immediate postpartum period. PMID- 17087893 TI - Real-time ultrasonic fetal head measurement and gestational age in dairy goats. AB - Dairy breed goat fetuses (n = 219) of known gestational age (Day 39 to Day 100) were examined transabdominally using a real-time ultrasound scanner with a 5 MHz linear-array transducer. Maximum head width was measured in symmetrical images using electronic linear distance calipers. This biparietal diameter (BPD) can be accurately measured as early as Day 40, but measurement becomes difficult after Day 105 of gestation due to the variability of fetal location and posture. The uniform, linear increase in BPD in the second trimester should allow for the accurate assignment of gestational age for doe confinement or induction of kidding. PMID- 17087894 TI - The effect of using the Breeding Soundness Evaluation as a selection criterion for Santa Gertrudis bulls on bulls in subsequent generations. AB - Results of Breeding Soundness Evaluations (BSE) used to select Santa Gertrudis bulls over a 10-yr period were analyzed in retrospect to determine the effect on bulls in subsequent generations. Bulls were tested at three ages: 16 mo (n = 1,936), 20 mo (n = 3,111) and at maturity (> or = 24 mo, n = 3,814). Scrotal circumference (SC) and sperm motility and morphology were evaluated. Sixteen-mo old bulls were required to have an SC > or = 30 cm to be tested. During 1977 to 1980, 20-mo-old bulls with an SC < 30 cm were culled, and the minimum SC was increased to 32 cm during 1981 to 1986. Mature bulls during this period were required to have an SC > or = 35 cm to be tested. A BSE < 60 was considered a failing score, and bulls were culled at this level. There was a positive correlation between SC and sperm motility and SC and sperm morphology in 16-mo old and 20-mo-old but not in mature bulls (P < 0.0001). The percentage of bulls passing the BSE was positively correlated with SC at all ages (P < 0.0001), and the passing rate increased over the years (P < 0.0001). Scrotal circumference increased over the years in bulls that passed the BSE (P < 0.0001). This increase with years was not as apparent in the semen quality traits measured. By using the BSE and a minimum SC to select prospective herd sires, the proportion of bulls passing the BSE is expected to increase in subsequent generations. Santa Gertrudis bulls can be reliably selected for use as herd sires at 16 mo of age. PMID- 17087895 TI - Survival of small and large littermate blastocysts in swine after synchronous and asynchronous transfer procedures. AB - Thirty-two crossbred sows were assigned to synchronous and asynchronous embryo transfer procedures to determine if, within a litter, small blastocysts were as viable as large blastocysts. Synchronous embryo transfers were established when donors and recipients displayed the onset of estrus (Day 0) within 6 h of each other. Asynchronous transfers were established when recipients displayed the onset of estrus 18 to 24 h after that of donors. An equal number (four or five) of the smallest and largest diameter blastocysts, from a Day 7 donor, were transferred to separate uterine horns of a Day 7 (synchronous) or a Day 6 (asynchronous) recipient. Each recipient's uterine horns were ligated at the external bifurcation to prevent transuterine embryonic migration. The percentage of blastocysts surviving was determined 300 h (12.5 d) after donors exhibited estrus. Small as well as large Day 7 blastocysts survived following asynchronos transfer to a Day 6 recipient. However, fewer (P<0.01) small blastocysts survived synchronous transfer than large blastocysts. These data suggested that small blastocysts were lost due to asynchrony with the uterine environment; however, when transferred to a less advanced environment, small blastocysts were equally viable as large blastocysts. PMID- 17087896 TI - Luteinizing hormone, testosterone and cortisol responses in rams upon presentation of estrous females in the nonbreeding season. AB - The physiological responses of luteinizing hormone, testosterone and cortisol in sexually experienced Ile de France rams to the introduction of estrous females were studied during the nonbreeding season. Blood sampling were collected from males for 7 h at 20-min intervals, starting 3 h before stimulation by estrous females. The differences in hormonal secretions were tested by comparisons between pretreatment and treatment Periods in 45 stimulated rams. Comparisons were conducted between rams that had increased LH pulse frequency and those that did not, between rams that ejaculated and those that did not, and between rams that were in direct physical contact and those that were kept at a distance of 30 cm from estrous females. Twenty-five rams (55% of the total) showed significant increases in LH pulse frequency (range, 0.80 to 4.00 peaks/ram/6 h, P<0.05), in basal and mean LH levels (1.5- and 2.5-fold, respectively), and in mean testosterone levels (3.5-fold). More frequent LH pulses had been found during the pretreatment period in 20 rams without increased LH pulse frequency. Eight ejaculating rams showed higher cortisol and mean, basal, and peak LH amplitude levels. Deprivation of physical contact with estrous females was associated with an absence of endocrine response. These results suggest that olfactory and/or tactile cues may be involved in the female effect on hormone levels. PMID- 17087897 TI - Disruption of pregnancy in rabbits serving as hosts for feeding ticks. AB - The effect of tick infestation on reproductive success of mammals was studied by using rabbits bred after they had served as hosts for Dermacentor variabilis . Mating success, pregnancy incidence, litter size and neonatal viability were reduced in tick-exposed rabbits, and the proportional number of abnormalities found in preimplantation-stage embryos was much higher than in unexposed controls. Fecundity was restored by prolonging the interval between tick exposure and breeding or by treatment with an acaricide. PMID- 17087898 TI - Synchronization of estrus in early diestral dairy heifers with Prostaglandin F(2)alpha and estradiol benzoate. AB - Following observation of estrus, 134 Holstein heifers were given injections of Prostaglandin F(2)alpha (PGF(2)alpha) between Days 5 and 10 of their cycle (estrus = Day 0). They were then randomly assigned to either a group receiving 400 microg of estradiol benzoate (E(2)B) 40 h or maintained as controls. Heifers observed in estrus within 120 h of PGF(2)alpha administration were inseminated (approximately 12 h after initial observation of estrus). Blood samples for progesterone determination were drawn from the coccygeal vein on Days 15 and 21 after insemination. Pregnancy was confirmed by palpation per rectum between Days 5.0 and 60 post insemination. When control and treated heifers were compared it was found that a higher percentage of heifers treated with E(2)B exhibited estrus after PGF(2)alpha, but there had been no effect on subsequent progesterone concentrations or pregnancy rates. PMID- 17087899 TI - Detecting estrus in synchronized heifers-using tailpaint and an aerosol raddle. AB - A synchronization treatment was initiated when each of 1227 heifers (four trials) was tailpainted. The tailpaint was sprayed with an aerosol raddle at the end of the treatment period. The heifers were in herds of 20 to 279 animals. Each herd was observed for estrus at selected post treatment intervals. A heifer was considered to be (or to have been) in estrus when the raddle was rubbed off. In three of the trials, animals which had the raddle removed were inseminated at 48 h following the end of the synchronization treatment. The tailpaint of an inseminated animal was scored from 0 (less than 10% of the paint remained) to 5 (more than 90% of the paint remained) and was then reraddled with a second color. The detection-insemination sequence was always repeated at 72 and 96 h, and sometimes at 120 h. Animals which had been previously inseminated, but then had paint scores reduced by at least 2 units were reinseminated 24 h later. Over the four trials, 94.5% of the heifers were detected in estrus through the use of the tailpaint and raddle system. The remaining 67 animals included only 10 (0.8%) which had ovulated without being detected in estrus. The reinsemination rate on consecutive days was 11.3% and was highest among animals that had a tailpaint score of 4 or 5 at 48 h. The proportion of animals detected in estrus at selected posttreatment intervals varied with the different synchronization treatments used within one herd, or with the same treatment used in different herds. The combination of tailpaint, raddling, tailpaint scoring and reraddling is a simple sequence which can be effectively used to detect estrus among heifers synchronized in research or commercial herds. PMID- 17087900 TI - Gonadotropin releasing hormone therapy in functional infertility of dairy cattle. AB - Effects of gonadotropin-releasing hormone (GnRH) on conception rate was tested in 379 repeat-breeders in nine large dairy herds in Louisiana. Cattle with three or more services were treated intramuscularly with GnRH at the time of artificial insemination. The conception rate for the repeat-breeders treated with GnRH was significantly greater than for the controls (56 vs 40%). Furthermore, repeat breeders that were treated with GnRH for two consecutive times at insemination resulted in a 53% increase in conception rate over the controls. PMID- 17087901 TI - Two cases of multiple abnormalities in testicular and ejaculated bull spermatozoa. AB - Multiple abnormalities were observed in testicular, epididynal and ejaculated spermatozoa from two bulls. These included acrosomal knobbing, incomplete nuclear condensation and coiled tails. The first two defects are considered to be characteristic of emmission of immature sperm, while the last may be a reflection of the effect of a changing osmotic environment on inherently susceptible cells. PMID- 17087902 TI - Reliability criteria of a bioassay using rats trained to detect estrus-specific odor in cow urine. AB - An odor discrimination apparatus was used to quantify the reaction of rats against odor differences of estrous and diestrous urine of cows. Rats were trained to release impulses from a microswitch when they came into contact with estrous but not diestrous urine. After training, the discrimination ability was highly significant (P< or = 0.001) between both urine types. The reaction was specific for cow urine, and no significant discrimination occurred for estrous and diestrous urine from pigs or goats, or that of women. The discrimination of urine diluted with water was still highly significant up to 1:25. Another group of rats was trained to detect camphor. Decreasing concentrations were detected down to 0.1/microg/ml when diluted in water, and down to 1/microg/ml when diluted in estrous urine. Once trained to discriminate between estrous and diestrous urine, rats maintained this ability for 12 mo. Thus, a bioassay was set up to accompany fractionation steps in the laboratory which could lead to the isolation of estrous pheromones in cow urine. PMID- 17087903 TI - Transcervical collection of equine conceptuses between 10 and 16 days after ovulation. AB - To recover intact Day-10.5 to Day-16.5 equine conceptuses (Day 0 = ovulation), a rigid catheter was used for 131 collections from donor mares diagnosed pregnant by ultrasonography. A total of 139 conceptuses were recovered, comprising 124 singletons, six pairs of twins and one set of triplets. Of these, 120 (86%) were intact after the collection, 14 (10%) had collapsed, and in five cases (4%), collapsed trophoblastic membranes were surrounded by an intact capsule. The recovery rate of intact conceptuses ranged from 99% on Days 10.5 to 12.5 to 40% on Day 16.5. More uterine flushes per recovery were needed to collect conceptuses on Day 14.5 than on Days 10.5 and 11.5 (x +/- SEM : 3.1 +/- 0.5 vs 1.4 +/- 0.1 and 1.3 +/- 0.2 flushes, respectively, P<0.05), and the total volume of flushing medium used was greater on Day 14.5 than on Days 10.5, 11.5 and 12.5 (1040 +/- 193 vs 406 +/- 49, 396 +/- 48 and 499 +/- 59 ml, respectively, P<0.01). Seventy of the 100 mares inseminated at the first estrus following embryo collection became pregnant, indicating that the technique used had no major effect on subsequent fertility. PMID- 17087904 TI - Decreased fertility in related females heterozygous for the 1/29 chromosome translocation. AB - Eighteen heifers and 120 cows which were descendants of a presumed 1/29 carrier Simmental bull were karotyped. Nine heifers (50%) and 48 cows (40%) were found to be heterozygous for the 1/29 translocation (59, XX, t(1q;29q)). The other animals were chromosomally normal (i.e., 60, XX) or not karotyped. The 48 1/29 cows were compared with 72 chromosomally normal cows with regards to days to first conception, calving interval, percentage of calves conceived, percentage of calves weaned and production efficiency (% calved conceived x % calved weaned). Nine carrier heifers were compared to the nine noncarrier heifers as to pregnancy status. Carrier, noncarrier and nonkarotyped relatives were compared to each other and to contemporary females with regard to pregnancy status at their initial exposure to males. The percentage of calves conceived (calving efficiency) in the 72 noncarrier and the 48 females heterozygous for the 1/29 translocation were 81.5 and 74.8%, respectively (P<0.07). Although days to first conception was longer and percentage of calves weaned and production efficiency were lower in the female heterozygous for the 1/29 translocation, the differences were not statistically different (P>0.10) from the noncarriers. Pregnancy rate was 44.4 and 66.7% (P>0.10) for nine carrier and nine noncarrier heifers, respectively. The pregnancy rate of carrier (65.4%), noncarrier (73.2%) and nonkarotyped (77.8%) relatives of this sire at their mating as yearlings, did not differ (P>0.10). The pregnancy rate as yearlings of carrier females (65.4%) and contemporary heifers (79.8%) did differ (P<0.05). Comparing the pregnancy rate as yearlings of all descendants (72.0%) of the Simmental sire to contemporary heifers (79.8%), a significant decrease (P<0.05) was found indicating that fertility of this sire may have been lower than other sires or that other factors beside the translocation affected fertility. PMID- 17087905 TI - Objective analysis of stallion sperm motility. AB - An image-analysis system utilizing a microcomputer and CellSoft computer-assisted semen analysis software package was evaluated to assess stallion sperm motility characteristics. Analyses were performed at 37 degrees C on a 6 microl drop of diluted semen placed on a glass slide and covered with an 18 mm(2) coverslip. Four groups of 25 cells each per slide, four slides per ejaculate and four ejaculates from each of three stallions were analyzed in a nested model. The percentage of motile sperm cells, mean velocity (microm/sec), mean linearity, and mean angular head displacement (microm) were measured. Statistical analysis of variance components showed that within ejaculates, more variation was accounted for in the differences among groups of 25 cells than among slides. Predicted standard deviations calculated for combinations of slides and groups of cells showed that a combination of two slides from which a total of 400 cells were analyzed resulted in a mean intra-assay coefficient of variation (CV) of 5.7% for the four measured variables. The following are individual coefficients of variation: percentage of motile cells (7.8%), mean velocity (6.4%), mean linearity (1.9%) and mean angular head displacement (6.6%). When ejaculate differences were included in the model and predicted standard deviations were calculated for a single ejaculate, the mean inter-assay CV was 9.2%. Mean velocity (6.4%) and mean linearity (4.7%) were more repeatable among ejaculates than either the percentage of motile sperm (14.4%) or angular head displacement (11.2%). It was concluded that this system is precise enough to determine differences in motility characteristics of stallion semen samples. PMID- 17087906 TI - B-mode ultrasound observations of bull testes during breeding soundness examinations. AB - The testes of 78 adult Beefmaster bulls (13 to > 31 mo old) were scanned with a 5 MHz, linear array portable ultrasound unit during routine breeding soundness examinations. Videotape recordings were used to measure mediastinum testis width and fluid width between visceral and parietal tunics (fluid width), and to count the number of fibrotic foci. Ultrasound results were compared to breeding soundness score parameters, which included total score, age classification, and percentage of primary and secondary sperm abnormalities. The overall mean mediastinum testis width and fluid width were 0.33 cm +/- 0.133 and 0.10 cm +/- 0.13, respectively; 15 testes had 1-8 fibrotic foci. There were no significant differences in mediastinum testis width and fluid width between left and right testes when compared by breeding soundness score classification or by age classification. There were no significant differences in mediastinum testis width, fluid width, breeding soundness score, or primary or secondary sperm abnormalities in bulls with fibrotic foci compared to bulls without fibrotic foci. There appears to be no significant information that routine testicular ultrasound examination adds to accepted breeding soundness evaluations performed on the same day. PMID- 17087907 TI - Some factors affecting successful vitrification of mouse blastocysts. AB - The effects of temperature and exposure time to vitrification solutions on In vitro survival of mouse blastocysts were investigated. Blastocysts were first exposed for 10 min to vitrification Solution 1 (VS1) containing 10% glycerol-20% 1,2 propanediol in phosphate buffered saline (PBS), then to vitrification Solution 2 (VS2) with 25 % glycerol-25% 1,2 propanediol for various periods either at room temperature or at 4 degrees C. At room temperature survival dropped quickly, while at 4 degrees C an increase in survival was observed. It is concluded that the viability of mouse blastocyts after vitrification is dependent on the temperature and duration of equilibration in vitrification solutions. PMID- 17087908 TI - Quality of semen stored at +15 degrees/16 degrees C as related to fertility of artificially inseminated swine. AB - Linear correlations between swine semen quality and fertility (birth rate and number live born pigs) were studied. Doses of swine semen with 4 x 10(9) motile spermatozoa were used after being stored at +15 degrees/16 degrees C . However, mean or high correlations could not be found. Low correlations occurred between head-acrosome anomalies, total anomalies and birth rate. It is likely that the doses of semen with 4 x 10(9) motile spermatozoa were too concentrated for the fertility level in the swine breed employed. PMID- 17087909 TI - Sex ratio of early embryos fertilized in vitro with spermatozoa separated by percoll. AB - Early bovine embryos were obtained by in vitro fertilization and sexing carried out by chromosome analysis. Separation of bovine X- and Y-bearing spermatozoa was performed using Percoll density gradient centrifugation and the enrichment of X sperm proportion was investigated. Through treatment with vinblastin sulfate and podophyllotoxin, 880 (48.6%) of 1812 embryos at two- to seven-cell stages at 48 to 53 h after sperm-egg incubation produced metaphase spreads, and 399 (45.3%) of these were successfully sexed; the sexable rate reaching 53.4% for four-cell embryos. Sexing rates for embryos from the original sperm of two bulls were 69.6% (32 46 ) in Bull A and 54.2% (58 107 ) in Bull B. Embryos fertilized in vitro with sperm sedimented at the bottom of sperm centrifuged under conditions (I) 50 to 85% of Percoll, 15 degrees C; (II) 30 to 80%, 10 degrees C; (III) 30 to 80% 20 degrees C; (IV) 30 to 90%, 20 degrees C, gave rise to male sex ratios of (I) 58.3% in Bull A and 53.5% in Bull B, (II) 65.9% in Bull A, (III) 49.3% in Bull B and (IV) 66.7% in Bull B. In conclusion, Percoll density gradient centrifugation under these four conditions was unsuccessful in separating X- and Y-bearing bull spermatozoa. PMID- 17087910 TI - Effect of ice nucleation by droplet of immobilized silver iodide on freezing of rabbit and bovine embryos. AB - Silver iodide was immobilized by applying the insoluble reaction between sodium alginate and calcium chloride. The immobilized silver iodide was immersed into a freezing solution in order to trigger ice nucleation. Temperature change during cooling and postthaw in vitro development of embryos were examined in order to evaluate the effectiveness of the immobilized silver iodide (AgI alginate-gel droplet) on embryo development. Samples containing the AgI alginate-gel droplets released the latent heat of fusion at a higher subzero temperature than samples without the AgI alginate-gel droplets. When the AgI alginate-gel droplet was added to the freezing solution of rabbit and bovine embryos, they were successfully preserved in liquid nitrogen. PMID- 17087911 TI - Seasonal variations in plasma hormones and reproductive efficiency in early postpartum buffalo. AB - Investigations were carried out on twenty newly calved Murrah buffalo that were divided into two sets of ten each during winter (December to February) and summer (May to September). Recommended feeding and management practices were followed. Blood samples were collected on Days 0, 1, 2, 4, 6, 8, 15, 22, 29, 36, 43, 50 and 57 postpartum for hormone radioimmune assay (RIA). The values of reproductive parameters and hormone levels during winter and summer, respectively, were the following for uterine involution, 39.50 +/- 2.74 d and 32.50 +/- 3.01 d; first postpartum estrus interval, 70.10 +/- 9.62 d and 37.40 +/- 11.82 d; number of services per conception, 1.57 +/- 0.29 and 2.40 +/- 0.68; first service conception rate, 40 and 20 %, overall conception rate, 70 and 40 %; 13,14-Dihydro 15-keto prostaglandin F(2alpha) (PGFM), 1.78 +/- 0.26 ng/ml and 2.62 +/- 0.30 ng/ml; progesterone, 1.09 +/- 0.14 ng/ml and 0.65 +/- 0.77 ng/ml. A negative correlation (r = -0.83, P 0.05 ) was observed between PGFM and progesterone. Although, summer stress hastens uterine involution and first postpartum estrus interval, overall reproductive efficiency is impaired due to deficiency of progesterone, which is essential for embryo survival. PMID- 17087912 TI - Estimating countermeasure effects for reducing collisions at highway-railway grade crossings. AB - Frequently transportation engineers are required to make difficult safety investment decisions in the face of uncertainty concerning the cost-effectiveness of different countermeasures. For certain types of highway-railway grade crossings, this problem is further aggravated due to the lack of observed before and after collision data that reflects the impact of specific countermeasures. This study proposes a Bayesian data fusion method as an attempt to overcome these challenges. In this framework, we make use of previous research findings on the effectiveness of a given countermeasure, which could vary by jurisdictions and operating conditions to obtain a priori inference on its expected effects. We then use locally calibrated models, which are valid for a specific jurisdiction, to develop the current best estimates regarding the countermeasure effects. By using a Bayesian framework, these two sources are integrated to obtain the posterior distribution of the countermeasure effectiveness. As a result, the outputs provide information not only of the expected collision response to a specific countermeasure but also its variance and corresponding probability distribution for a range of likely values. Examples from Canadian highway-railway grade crossing data are used to illustrate the proposed methodology and the specific effects of prior knowledge and data likelihood on the combined estimates of countermeasure effects. PMID- 17087913 TI - Chemiluminescence-based detection and comparison of protein amounts adsorbed on differently modified silica surfaces. AB - The biological consequences of protein adsorption on biomaterial surfaces are considered to be of utmost importance for their biocompatibility. A new method based on amino group-labeling coupled to a chemiluminescence reaction for direct determination of proteins adsorbed on material surfaces was employed. This method was used to explore the effects of surface chemistry and surface roughness on protein adsorption in a silicon oxide model system. Corundum sandblasting was applied to silicon wafers to create roughened surfaces while immobilization of fluorocarbon-, hydrocarbon-, and poly(ethylene glycol)-containing silanes produced surfaces of varying wettability. The adsorption behavior of two complex body fluids, human serum and saliva, and of two purified components, human serum albumin and fibronectin, was strongly influenced by the surface parameters. A general tendency to higher amounts of adsorbed protein was found on roughened surfaces and modification with poly(ethylene glycol) or with fluorocarbon moieties reduced protein adsorption. The values obtained with the new method could be confirmed by a colorimetric determination of protein amounts adsorbed on identically modified silica beads and were in accordance with those previously reported utilizing established methods for protein quantification. The presented method, which was methodically simple to perform and allowed the simultaneous measurement of a large number of samples, may be of future value for high throughput surveying of the protein adsorption characteristics of biomaterials. PMID- 17087914 TI - Heartbeat perception in depression. AB - Alterations in bodily awareness have been implicated in depression but there has been little detailed empirical characterisation of the degree and accuracy of body perception in the disorder. The present study examined the objective accuracy of heartbeat perception (using the Schandry mental tracking task) and the subjective degree of bodily focus (using the Bodily Consciousness Questionnaire; BCQ) in healthy control volunteers, a moderately depressed community sample, and a more severely depressed clinic sample (n=18 in each group). The community sample showed less accurate heartbeat perception than the control group as expected. Counter to prediction, however, the more severely depressed clinic sample performed better than the community depressed sample and equivalently to control volunteers on the Schandry task. There were no group differences on subjective bodily awareness. Implications for theories of depression are discussed. PMID- 17087915 TI - The cognitive mediation of thought-control strategies. AB - Research suggests that people use various strategies to control their normally occurring intrusive thoughts. Strategies that involve worrying about the thought and self punishment appear to be associated with certain forms of psychopathology, such as obsessive-compulsive (OC) symptoms. The present study sought to examine whether dysfunctional beliefs associated with OC symptoms (e.g., beliefs that intrusive thoughts are highly significant) underlie the use of such thought control strategies. Ninety-three non-clinical participants completed self-report questionnaires measuring cognitive variables, thought control strategies, and OC symptoms. Analyses revealed that overestimates of threat and responsibility, beliefs about the significance and need to control intrusions, the need for perfection and certainty, and scrupulosity were associated with the use of punishment, but not worry thought control strategies. These cognitive phenomena also mediated the relationship between OC symptoms and the use of punishment as a thought-control strategy. Results are discussed in terms of cognitive models of OC symptoms and their implications for cognitive behavioral therapy. PMID- 17087916 TI - Combination of integrin siRNA and irradiation for breast cancer therapy. AB - Up-regulation of integrin alpha(v)beta(3) has been shown to play a key role in tumor angiogenesis and metastasis. In this study, we evaluated the role of integrin alpha(v)beta(3) in breast cancer cell resistance to ionizing irradiation (IR) and tested the anti-tumor efficacy of combining integrin alpha(v) siRNA and IR. Colonogenic survival assay, cell proliferation, apoptosis, and cell cycle analysis were carried out to determine the treatment effect of siRNA, IR, or combination of both on MDA-MB-435 cells (integrin alpha(v)beta(3)-positive). Integrin alpha(v)beta(3)-negative MCF-7 cells exert more radiosensitivity than MDA-MB-435 cells. IR up-regulates integrin alpha(v)beta(3) expression in MDA-MB 435 cells and integrin alpha(v) siRNA can effectively reduce both alpha(v) and alpha(v)beta(3) integrin expression, leading to increased radiosensitivity. Integrin alpha(v) siRNA also promotes IR-induced apoptosis and enhances IR induced G2/M arrest in cell cycle progression. This study, with further optimization, may provide a simple and highly efficient treatment strategy for breast cancer as well as other integrin alpha(v)beta(3)-positive cancer types. PMID- 17087917 TI - Mitochondrial ROS-PKCepsilon signaling axis is uniquely involved in hypoxic increase in [Ca2+]i in pulmonary artery smooth muscle cells. AB - The molecular mechanisms underlying hypoxic responses in pulmonary and systemic arteries remain obscure. Here we for the first time report that acute hypoxia significantly increased total PKC and PKCepsilon activity in pulmonary, but not mesenteric arteries, while these two tissues showed comparable PKCepsilon protein expression and activation by the PKC activator phorbol 12-myristate 13-acetate. Hypoxia induced an increase in intracellular reactive oxygen species (ROS) generation in isolated pulmonary artery smooth muscle cells (PASMCs), but not in mesenteric artery SMCs. Inhibition of mitochondrial ROS generation with rotenone, myxothiazol, or glutathione peroxidase-1 overexpression prevented hypoxia-induced increases in total PKC and PKCepsilon activity in pulmonary arteries. The inhibitory effects of rotenone were reversed by exogenous hydrogen peroxide. A PKCepsilon translocation peptide inhibitor or PKCepsilon gene deletion decreased hypoxic increase in [Ca(2+)](i) in PASMCs, whereas the conventional PKC inhibitor GO6976 had no effect. These data suggest that acute hypoxia may specifically increase mitochondrial ROS generation, which subsequently activates PKC, particularly PKCepsilon, contributing to hypoxia-induced increase in [Ca(2+)](i) and contraction in PASMCs. PMID- 17087918 TI - Dissociating neural indices of dynamic cognitive control in advance task-set preparation: an ERP study of task switching. AB - Switching between different tasks is associated with performance deficits, or 'switch costs', relative to repeating the same task. Recent evidence suggests that response rather than task selection processes may be a major cause of switch costs [Schuch, S., Koch, I., 2003. The role of response selection for inhibition of task sets in task shifting. J Exp Psychol Hum Percept Perform, 29 (1) 92-105]. Thus, switch costs are not incurred if on the preceding trial a task has been prepared for but no response required (a 'no-go' trial). We investigated the relationship between response selection and the subsequent preparation of an alternative task set. While switch costs were absent following 'no-go' trials, ERP differences during the precueing interval showed that response selection has implications for subsequent task preparation as well as for task performance per se. The results are discussed in relation to the dissociation of intention versus action in behavioural control and the role of inhibition in switching between task sets. PMID- 17087919 TI - Alpha-synuclein enhances dopamine D2 receptor signaling. AB - Parkinson's disease (PD) is characterized by a selective loss of dopamine producing neurons in the substantia nigra (SN), which in turn results in dopamine depletion in the striatum, and the presence of neuronal cytoplasmic inclusions known as Lewy bodies (LBs). Alpha-synuclein is a presynaptic protein that accumulates abnormally in LBs and is seen predominantly in cases of dementia with LBs. Although the central role of alpha-synuclein in neurodegeneration has been previously demonstrated by the discovery of missense alpha-synuclein mutations in familial PD, the specific mechanism by which alpha-synuclein contributes to these diseases remains unclear. In the present study, we examined whether alpha synuclein affects the downstream signaling of dopamine D2 receptor (D2R). In CHO cells stably transfected with D2Rs, alpha-synuclein enhanced dopamine D2-agonist mediated inhibition of adenylate cyclase, which consequently affected its downstream cAMP-responsive element (CRE)-mediated gene transcription, while C terminal deletion mutant of alpha-synuclein did not. Our study suggests that the alpha-synuclein enhances the dopamine-mediated intracellular signaling pathways by D2R, thus provide a possible mechanism in presynaptic regulation of the synaptic homeostasis in the dopaminergic neurotransmission. PMID- 17087920 TI - Sex, genes and steroids. Proceedings of the Society for Women's Health Research Sex and Gene Expression Conference in conjunction with the Workshop on Steroid Hormones and Brain Function. March 28-April 1, 2006. Breckenridge, Colorado, USA. PMID- 17087922 TI - Increased sensitivity of chitosan determination by a dye binding method. AB - Chitosan is a topic of current research in pharmaceutics, medicine, biotechnology, and beyond. This note describes an improved quantification of chitosan using the dye Cibacron Brilliant Red 3B-A. The method is sensitive and of a good reproducibility and linearity in the range of 10-80 microg/mL. PMID- 17087921 TI - Attentional modulation of early-stage visual processing in schizophrenia. AB - This study shows that paying attention to the color of a visual stimulus is manifested by an early endogenous scalp-positive event-related brain potential (ERP) component, referred to as "selection positivity", that emerges within the first 100 ms after stimulus onset in healthy observers. In contrast, recently ill and chronically ill schizophrenia patients as well as patients at high risk for schizophrenia all failed to show this early ERP component while attending to color. These results suggest that a relatively early stage of visual-selective processing in posterior extrastriate cortex is disrupted in schizophrenia. PMID- 17087923 TI - Direct oxidation of sugar nucleotides to the corresponding uronic acids: TEMPO and platinum-based procedures. AB - The direct oxidation of UDP-alpha-d-glucose and UDP-N-acetyl-alpha-d-glucosamine to the corresponding uronic acids was explored using either TEMPO or platinum catalysed oxidation with molecular oxygen. Whilst TEMPO-based procedures gave rise to substantial over-oxidation and/or degradation of UDP-glucose, oxidation of UDP-N-acetyl-glucosamine to UDP-N-acetyl-glucosaminuronic acid was achieved with >90% conversion and ca. 65% isolated yield using a platinum-catalysed procedure. PMID- 17087924 TI - Nucleophilic substitution at the anomeric position of 1,2-O isopropylidenefuranose derivatives. A novel stereoselective synthesis of cyclic phosphates analogous to cAMP. AB - 1,2-O-Isopropylidenefuranose derivatives were treated with various nucleophiles in the presence of either BF(3).OEt(2) or trimethylsilyl trifluoromethanesulfonate (TMSOTf) leading to substitution products in a regio- and stereoselective manner. In particular, nucleophilic substitution of 1,2-O isopropylidenefuranose derivatives when treated with allyltrimethylsilane was controlled by steric and electronic factors (similar to Woerpel's stereoelectronic model). On the other hand, when 1,2-O-isopropylidenefuranose derivatives were treated with trimethylsilane, in the presence of bis-O trimethylsilyl-5-iodouracil or bis-O-trimethylsilyl-thymidine, substitution products were generated in high regio- and stereoselectivities via an unusual nucleophilic substitution with opening of the furanose ring. Based on these results, a stereoselective method for the synthesis of neutral cyclic phosphates analogous to cAMP was developed. PMID- 17087925 TI - Crystal and molecular structures of diglycosyl disulfide derivatives. AB - The crystal and molecular structures of S-(2,3,4,6-tetra-O-acetyl-beta-d glucopyranosyl)-S'-(2,3,4,6-tetra-O-acetyl-beta-d-galactopyranosyl)disulfide (1) and the mono-sulfoxide (3) of bis(2,3,4,6-tetra-O-acetyl-beta-d glucopyranosyl)disulfide (2) are described. Comparison of 2 and 3, containing -S S- and -S-S(O)- bonds, respectively, allows to delineate structural differences brought about by the different oxidation states of one of the sulfur atoms in carbohydrate disulfides. PMID- 17087926 TI - O-Acetylation in the O-specific polysaccharide isolated from Shigella flexneri serotype 2a. AB - Shigella flexneri causes diarrheal diseases especially in infants and children in developing countries. Modifications of the lipopolysaccharide (LPS) molecule, like bacteriophage-mediated glucosylation and acetylation of the O-specific chain (O-SP), are important for the LPS antigenicity and consequently for the immunogenicity of the polysaccharide-based vaccines against shigellosis. Here, we report the degree of O-acetylation and the localisation of O-acetyl groups and side-chain glucose substitution in the O-SP (scheme) in different preparations of S. flexneri type 2a LPS. [structure: see text] PMID- 17087927 TI - CD4+ lymphocytes require platelets for adhesion to immobilized fibronectin in flow: role of beta(1) (CD29)-, beta(2) (CD18)-related integrins and non-integrin receptors. AB - The role of platelets in T-lymphocytes adhesion is not clear yet. Herpesvirus saimiri (HVS)-infected CD4(+) T-lymphocytes were placed into polystyrene plates pre-coated with fibronectin. The adherent T-cells were enumerated by image analysis. Under static condition, 38+/-10cells/mm(2) adhered and addition of gel filtered platelets (GFP) and PMA enhanced cell adhesion 4.3- and 4.1-fold. Using PMA plus GFP 11.9-fold enhancement in cell adhesion was achieved. In contrast, under flow (200s(-1)), neither basal adhesion nor following separate addition of PMA or GFP was observed, whereas combined addition of PMA and GFP induced noticeable adhesion (34cells/mm(2)). The adhesion was inhibited by blockade of alpha(5)-integrin (CD49e, 87%), beta(2)-integrin (CD18, 78%), CD40L (60%), PSGL-1 (CD162, 60%), and CD40L plus PSGL-1 (83%). Thus, activated platelets promote activated T-cell adhesion to fibronectin under flow via integrins (alpha(5)beta(1), and alpha(L)beta(2)), CD40-CD40L and P-selectin-PSGL-1 mediated interactions. PMID- 17087928 TI - Protein tyrosine and serine-threonine phosphatases in the sea urchin, Strongylocentrotus purpuratus: identification and potential functions. AB - Protein phosphatases, in coordination with protein kinases, play crucial roles in regulation of signaling pathways. To identify protein tyrosine phosphatases (PTPs) and serine-threonine (ser-thr) phosphatases in the Strongylocentrotus purpuratus genome, 179 annotated sequences were studied (122 PTPs, 57 ser-thr phosphatases). Sequence analysis identified 91 phosphatases (33 conventional PTPs, 31 dual specificity phosphatases, 1 Class III Cysteine-based PTP, 1 Asp based PTP, and 25 ser-thr phosphatases). Using catalytic sites, levels of conservation and constraint in amino acid sequence were examined. Nine of 25 receptor PTPs (RPTPs) corresponded to human, nematode, or fly homologues. Domain structure revealed that sea urchin-specific RPTPs including two, PTPRLec and PTPRscav, may act in immune defense. Embryonic transcription of each phosphatase was recorded from a high-density oligonucleotide tiling microarray experiment. Most RPTPs are expressed at very low levels, whereas nonreceptor PTPs (NRPTPs) are generally expressed at moderate levels. High expression was detected in MAP kinase phosphatases (MKPs) and numerous ser-thr phosphatases. For several expressed NRPTPs, MKPs, and ser-thr phosphatases, morpholino antisense-mediated knockdowns were performed and phenotypes obtained. Finally, to assess roles of annotated phosphatases in endomesoderm formation, a literature review of phosphatase functions in model organisms was superimposed on sea urchin developmental pathways to predict areas of functional activity. PMID- 17087929 TI - In vitro and in vivo interaction of synthetic peroxide RBx11160 (OZ277) with piperaquine in Plasmodium models. AB - RBx11160 (OZ277) is a promising antimalarial drug candidate that Ranbaxy Laboratories Limited and Medicines for Malaria Venture (MMV) are currently developing as a fixed combination with piperaquine. Here, we describe the in vitro (Plasmodium falciparum) and in vivo (Plasmodium berghei) activities of piperaquine in combination with RBx11160 and artemether. In vitro, both combinations demonstrated a slight tendency towards antagonism with mean sums of fractional inhibitory concentrations (mean Sigma FICs) of 1.5. In vivo, piperaquine and artemether were borderline antagonistic (mean Sigma FIC of 1.4). However, an additive in vivo interaction of piperaquine and RBx11160 (mean Sigma FIC of 1.1) was identified, suggesting that a RBx11160-piperaquine combination therapy in humans should allow each molecule to exert its full antimalarial effect. PMID- 17087930 TI - Leishmania infantum: mixed T-helper-1/T-helper-2 immune response in experimentally infected BALB/c mice. AB - The main goal of the present study was to characterise the course of infection and immunological responses developed by Leishmania infantum infected BALB/c mice. Parasite load was determined by Real-time TaqMan PCR while cytokine and Immunoglobulin G (IgG) production were assessed by ELISA. Leishmania DNA was detected in spleen and liver as soon as day 1 post-inoculation (pi) and the parasitism was sustained until the end of the experiment. The cytokine kinetics in spleen and liver was generally associated with the oscillations of parasite load. Overall, it was not observed a distinct Th1 or Th2 pattern of cytokine production during the time of experiment. The infected mice developed a mixed immune response, with concomitant production of IFN-gamma, IL-4 and IL-10, both in spleen and liver, and both IgG isotypes. However, our results suggest that, compared to liver, the spleen is more susceptible to L. infantum infection. PMID- 17087931 TI - Recurrent genomic alterations with impact on survival in colorectal cancer identified by genome-wide array comparative genomic hybridization. AB - BACKGROUND & AIMS: Although genetic aspects of tumorigenesis in colorectal cancer (CRC) have been well studied, reliable biomarkers predicting prognosis are scarce. We aimed to identify recurrently altered genomic regions (RAR) in CRC with high resolution, to investigate their implications on survival and to explore novel cancer-related genes in prognosis-associated RARs. METHODS: A 1-Mb resolution microarray-based comparative genomic hybridization (array CGH) was applied to 59 CRCs. RARs, defined as genomic alterations, detected in more than 10 cases were identified and analyzed for their association with survival. Expression levels of genes in prognosis-associated RARs were examined by real time quantitative polymerase chain reaction. RESULTS: Twenty-seven RARs were identified. Eleven high-level amplifications and 2 homozygous deletions also were detected, but they were not as common as RARs. Multivariate analysis revealed RAR L1 (loss on 1p36; hazard ratio = 8.15, P = .002) and RAR-L20 (loss on 21q22; hazard ratio = 3.53, P = .034) are independent indicators of poor prognosis. Expression of CAMTA1, located in RAR-L1, was reduced frequently in CRCs, and low CAMTA1 expression was associated significantly with poor prognosis, which indicates that CAMTA1 may play a role as a tumor suppressor in CRC. Five pairs of RARs were correlated significantly to each other and 3 pairs share genes involved in the same biological functions, suggesting possible collaborative roles in tumorigenesis. CONCLUSIONS: We identified recurrent genomic changes in 59 CRCs. RARs could be more important in sporadic tumors where the effect of genomic changes on tumorigenesis is relatively smaller than in familial cancer. Our results and analysis strategy will be helpful to elucidate pathogenesis of CRCs or to develop biomarkers for predicting prognosis. PMID- 17087932 TI - Microsatellite instability in interval colon cancers. AB - BACKGROUND & AIMS: Colon cancers that develop after a complete colonoscopy may be the result of "failure of colonoscopy" or rapid tumor growth. Tumors that develop via the mismatch repair gene pathway demonstrate rapid tumor growth. The aim of this study was to determine if interval colon cancers were more likely than noninterval cancers to result from the loss of function of mismatch repair genes and hence demonstrate microsatellite instability (MSI). METHODS: We searched our institution's cancer registry for interval cancers, defined as colon cancers that developed within 5 years of a complete colonoscopy. These were frequency matched in a 1:2 ratio by age and sex to patients with noninterval cancers (defined as colon cancers diagnosed on a patient's first recorded colonoscopy). Archived cancer specimens for all subjects were retrieved and tested for MSI. RESULTS: Of the 993 colon cancers diagnosed during the study period, 51 (5.1%) were identified as an interval cancer, and 112 subjects with noninterval cancer served as a comparison group. Study subjects were almost all men. MSI was found in 30.4% of interval cancers compared with 10.3% of noninterval cancers (P = .003). After adjusting for age, interval cancers were 3.7 times more likely to show MSI than noninterval cancers (95% confidence interval, 1.5-9.1). This association was strongest for tumors located in the distal colon (odds ratio, 17.5; P = .008). No difference in TNM stage at diagnosis, histologic type or grade, or 5-year survival was found between groups. CONCLUSIONS: Interval colon cancers were almost 4 times as likely as noninterval colon cancers to be associated with mismatch repair gene dysfunction. PMID- 17087933 TI - A mouse model of hereditary pancreatitis generated by transgenic expression of R122H trypsinogen. AB - BACKGROUND & AIMS: Missense mutations in human cationic trypsinogen PRSS1 are frequently detected in patients with hereditary pancreatitis, a rare genetic disease of the pancreas characterized by autodigestive necrosis, chronic inflammation, and fibrosis. To examine the link between PRSS1 mutations and the initiation and progression of hereditary pancreatitis, we have sought to generate a transgenic mouse that carries a missense mutation in the PRSS1 that is most frequently observed in patients. METHODS: A transgenic mouse was generated in which the expression of the mouse PRSS1 mutant R122H (R122H_mPRSS1) is targeted to pancreatic acinar cells by fusion to the elastase promoter. The expression of the mutant trypsinogen was assessed by immunohistochemical staining and real-time reverse transcription polymerase chain reaction analysis. The relationship between transgene expression and inflammation was analyzed by morphologic assessment of H&E-stained tissue sections, responsiveness to cerulein-induced pancreatitis, and immunohistochemical identification of cellular and biochemical components of the inflammatory response. RESULTS: Pancreata from transgenic mice display early-onset acinar cell injury and inflammatory cell infiltration. With progressing age, the transgenic mice develop pancreatic fibrosis and display acinar cell dedifferentiation. Moreover, the expression of R122H_mPRSS1 transgene is associated with enhanced response to cerulein-induced pancreatitis. Finally, cell-specific activation of the inflammation-associated signaling pathways, c-jun N-terminal kinase and extracellular signal-regulated kinase, was observed in response to expression of R122H_mPRSS1. CONCLUSIONS: These results underscore the importance of PRSS1 mutations as pathogenic mediators of hereditary pancreatitis and indicate that persistent pancreatic injury might be causally linked to chronic pancreatitis. PMID- 17087934 TI - Computed tomographic colonography: assessment of radiologist performance with and without computer-aided detection. AB - BACKGROUND & AIMS: In isolation, computer-aided detection (CAD) for computed tomographic (CT) colonography is as effective as optical colonoscopy for detection of significant adenomas. However, the unavoidable interaction between CAD and the reader has not been addressed. METHODS: Ten readers trained in CT but without special expertise in colonography interpreted CT colonography images of 107 patients (60 with 142 polyps), first without CAD and then with CAD after temporal separation of 2 months. Per-patient and per-polyp detection were determined by comparing responses with known patient status. RESULTS: With CAD, 41 (68%; 95% confidence interval [CI], 55%-80%) of the 60 patients with polyps were identified more frequently by readers. Per-patient sensitivity increased significantly in 70% of readers, while specificity dropped significantly in only one. Polyp detection increased significantly with CAD; on average, 12 more polyps were detected by each reader (9.1%, 95% CI, 5.2%-12.8%). Small- (< or =5 mm) and medium-sized (6-9 mm) polyps were significantly more likely to be detected when prompted correctly by CAD. However, overall performance was relatively poor; even with CAD, on average readers detected only 10 polyps (51.0%) > or =10 mm and 24 (38.2%) > or =6 mm. Interpretation time was shortened significantly with CAD: by 1.9 minutes (95% CI, 1.4-2.4 minutes) for patients with polyps and by 2.9 minutes (95% CI, 2.5-3.3 minutes) for patients without. Overall, 9 readers (90%) benefited significantly from CAD, either by increased sensitivity and/or by reduced interpretation time. CONCLUSIONS: CAD for CT colonography significantly increases per-patient and per-polyp detection and significantly reduces interpretation times but cannot substitute for adequate training. PMID- 17087935 TI - AGA Institute Medical Position Statement on the Diagnosis and Management of Celiac Disease. PMID- 17087936 TI - Effects of PKC412, nilotinib, and imatinib against GIST-associated PDGFRA mutants with differential imatinib sensitivity. AB - BACKGROUND & AIMS: Activating mutations in platelet-derived growth factor receptor alpha (PDGFRA) have been reported in a subset of gastrointestinal stromal tumor (GIST) patients who do not express the mutant stem cell factor receptor c-kit. The responsiveness of mutant PDGFRA-positive GIST to imatinib depends on the location of the PDGFRA mutation; for example, the V561D juxtamembrane domain mutation is more sensitive to imatinib than the D842V kinase domain mutation. In this study, we compare the effects of 3 tyrosine kinase inhibitors, PKC412 and nilotinib, and imatinib, on 2 GIST-related PDGFRA mutants, V561D and D842V, which possess differential sensitivity to imatinib. METHODS: The effects of PKC412, nilotinib, and imatinib, alone and in combination, were evaluated via in vitro proliferation studies performed with V561D- or D842V PDGFRA mutants. The effects of nilotinib and PKC412, alone and combined, were investigated in vivo. RESULTS: PKC412 potently inhibited the V561D-PDGFRA mutant in vitro and the D842V-PDGFRA mutant in vitro and in vivo. Both imatinib and nilotinib displayed potent activity in vitro against the V561D-PDGFRA mutant but were significantly less efficacious against D842V-PDGFRA. However, when combined with either imatinib or PKC412, nilotinib showed no evidence for antagonism and acted in a cooperative fashion against D842V-PDGFRA. CONCLUSIONS: Our findings support the clinical testing of PKC412 for treatment of mutant PDGFRA-GIST. The data also support the use of nilotinib as a treatment option for V561D-PDGFRA associated GIST, although the reduced sensitivity of D842V-PDGFRA probably limits the potential of nilotinib monotherapy for D842V-PDGFRA-associated GIST. PMID- 17087937 TI - American Gastroenterological Association (AGA) Institute technical review on the diagnosis and management of celiac disease. PMID- 17087938 TI - A molecular signature to discriminate dysplastic nodules from early hepatocellular carcinoma in HCV cirrhosis. AB - BACKGROUND & AIMS: Small liver nodules approximately 2 cm are difficult to characterize by radiologic or pathologic examination. Our aim was to identify a molecular signature to diagnose early hepatocellular carcinoma (HCC). METHODS: The transcriptional profiles of 55 candidate genes were assessed by quantitative real-time reverse-transcription polymerase chain reaction (RT-PCR) in 17 dysplastic nodules (diameter, 10 mm) and 20 early HCC (diameter, 18 mm) from HCV cirrhotic patients undergoing resection/transplantation and 10 nontumoral cirrhotic tissues and 10 normal liver tissues. Candidate genes were confirmed by quantitative RT-PCR in 20 advanced HCCs and by immunohistochemistry in 75 samples and validated in an independent set of 29 samples (dysplastic nodules [10] and small HCC [19; diameter, 20 mm]). RESULTS: Twelve genes were significantly, differentially expressed in early HCCs compared with dysplastic nodules (>2-fold change; area under the receiver operating characteristic curve > or =0.8): this included TERT, GPC3, gankyrin, survivin, TOP2A, LYVE1, E-cadherin, IGFBP3, PDGFRA, TGFA, cyclin D1, and HGF. Logistic regression analysis identified a 3 gene set including GPC3 (18-fold increase in HCC, P = .01), LYVE1 (12-fold decrease in HCC, P = .0001), and survivin (2.2-fold increase in HCC, P = .02), which had a discriminative accuracy of 94%. The validity of the gene signature was confirmed in a prospective testing set. GPC3 immunostaining was positive in all HCCs and negative in dysplastic nodules (22/22 vs 0/14, respectively, P < .001). Nuclear staining for survivin was positive in 12 of 13 advanced HCC cases and in 1 of 9 early tumors. CONCLUSIONS: Molecular data based on gene transcriptional profiles of a 3-gene set allow a reliable diagnosis of early HCC. Immunostaining of GPC3 confirms the diagnosis of HCC. PMID- 17087939 TI - Inhibition of Smad7 with a specific antisense oligonucleotide facilitates TGF beta1-mediated suppression of colitis. AB - BACKGROUND & AIMS: Defective transforming growth factor (TGF)-beta1 signaling due to high levels of Smad7 is a feature of inflammatory bowel disease (IBD). In this study, we analyzed the effect of reducing Smad7 levels with antisense oligonucleotide on mouse models of colitis. METHODS: Mucosal samples taken from colitic tissue of mice with colitis due to either haptenating reagents (trinitrobenzene sulfonic acid [TNBS] or oxazolone) or to transfer of T cells (SCID transfer colitis) were analyzed for Smad3 and/or Smad7 expression by Western blotting and, in some cases, content of TGF-beta1 by enzyme-linked immunosorbent assay. The effect of oral Smad7 antisense oligonucleotide on mucosal inflammation was assessed. RESULTS: TGF-beta1 levels were increased in the inflamed tissues of mice with colitis induced by either TNBS or oxazolone. Nevertheless, TGF-beta1 did not exert a regulatory effect, probably because TGF beta1 signaling was blocked, as indicated by the presence of reduced Smad3 phosphorylation and high levels of Smad7. Oral administration of Smad7 antisense oligonucleotide to colitic mice restored TGF-beta1 signaling via Smad3 and ameliorated inflammation in hapten-induced colitis. In addition, Smad7 antisense oligonucleotide had a therapeutic effect on relapsing TNBS-induced colitis but not on cell-transfer colitis. CONCLUSIONS: These data suggest that colitis models associated with high endogenous TGF-beta1 levels and defective TGF-beta1 signaling due to high levels of Smad7 can be ameliorated by down-regulation of Smad7 and by oral administration of Smad7 antisense oligonucleotide. This may represent a new approach to the control of IBD, particularly during active phases when its Smad7 profile resembles that of hapten-induced colitis. PMID- 17087940 TI - Genetic variation in myosin IXB is associated with ulcerative colitis. AB - BACKGROUND & AIMS: Common germline genetic variation in the 3' region of myosin IXB (MYO9B) has been associated recently with susceptibility to celiac disease, with a hypothesis that MYO9B variants might influence intestinal permeability. These findings suggested the current study investigating a possible further role for MYO9B variation in inflammatory bowel disease. METHODS: Eight single nucleotide polymorphisms (SNPs) were selected to tag common haplotypes from the 35-kb 3' region of MYO9B. These included the strongest celiac disease-associated variants reported in a Dutch cohort. These SNPs were studied in 3 independently collected and genotyped case-control cohorts of European descent (UK, Dutch, and Canadian/Italian), comprising in total 2717 inflammatory bowel disease patients (1197 with Crohn's disease, 1520 with ulcerative colitis) and 4440 controls. RESULTS: Common variation in MYO9B was associated with susceptibility to inflammatory bowel disease in all 3 cohorts examined (most associated SNP, rs1545620; meta-analysis P = 1.9 x 10(-6); odds ratio, 1.2), with the same alleles showing association as reported for celiac disease. CONCLUSIONS: MYO9B genetic variants predispose to inflammatory bowel disease. Interestingly, rs1545620 is a nonsynonymous variant leading to an amino acid change (Ala1011Ser) in the third calmodulin binding IQ domain of MYO9B. Unlike previous variants (in other genes) reported to predispose to inflammatory bowel disease, the association at MYO9B was considerably stronger with ulcerative colitis, although weaker association with Crohn's disease also was observed. These data imply shared causal mechanisms underlying intestinal inflammatory diseases. PMID- 17087941 TI - Novel model of antigen-specific induction of bile duct injury. AB - BACKGROUND & AIMS: Biliary-directed inflammation is an important cause of acute and chronic liver disease. We developed and characterized a transgenic mouse model of immune-mediated hepatobiliary injury. METHODS: Ovalbumin (OVA)-BIL mice were developed using 3.0 kilobase of the rat apical sodium-dependent bile acid transporter promoter to drive aberrant expression of a membrane form of ovalbumin (OVA) on biliary epithelium. Liver inflammation resulted from adoptive transfer of OVA-specific T cells. Liver immune cells were characterized to determine the mechanism of the response by assessing activation, proliferation, and intracellular cytokine expression. RESULTS: OVA-BIL transgenic mice were tolerant to OVA, without evidence of liver disease. Adoptive transfer of OVA-specific CD4+ and CD8+ T cells into naive OVA-BIL mice led to biliary-centered necroinflammatory damage in a dose-dependent manner. This inflammation absolutely required CD8+ T cells and was augmented by CD4+ T cells. Adoptively transferred OVA CD8+ cells homed to and proliferated in the liver but not the spleen. These activated, adoptively transferred cytotoxic T lymphocytes produced elevated levels of tumor necrosis factor alpha and interferon gamma. CONCLUSIONS: T-cell recognition of antigen aberrantly expressed on bile duct epithelium induced an acute necroinflammatory response specific to the liver, with activation, proliferation, and cytokine production predominantly by the OVA-specific cytotoxic T cells. Thus, OVA BIL represents an antigen-specific animal model of inflammatory bile duct injury. PMID- 17087942 TI - The CpG island methylator phenotype and chromosomal instability are inversely correlated in sporadic colorectal cancer. AB - BACKGROUND & AIMS: The CpG island methylator phenotype (CIMP) is one of the mechanisms involved in colorectal carcinogenesis (CRC). Although CIMP is probably the cause of high-frequency microsatellite instability (MSI-H) sporadic CRCs, its role in microsatellite stable (MSS) tumors is debated. The majority of MSS CRCs demonstrate chromosomal instability (CIN) with frequent loss of heterozygosity (LOH) at key tumor suppressor genes. We hypothesized that the majority of sporadic CRCs without CIN would be associated with CIMP. METHODS: We tested 126 sporadic CRCs for MSI and LOH and categorized tumors into MSI, LOH, or MSI-/LOH- subgroups. Methylation status was evaluated using 6 CIMP-related markers (MINT1, MINT2, MINT31, p16(INK4alpha), p14(ARF), and hMLH1) and 6 tumor suppressor genes (PTEN, TIMP3, RUNX3, HIC1, APC, and RARbeta2). BRAF V600E mutation analysis was performed using allele-specific polymerase chain reaction and DNA sequencing. RESULTS: We observed frequent methylation at all 12 loci in all CRCs. BRAF V600E mutations correlated with the MSI (P < .0001) and MSI-/LOH- (P = .03) subgroups. MSI and MSI-/LOH- tumors exhibited more promoter methylation than CRCs with LOH (P < .0001). We also found an inverse correlation between the frequencies of methylation and LOH (rho = -0.36; P < .0001). CONCLUSIONS: The associations between methylation frequencies at CIMP-related markers and MSI or MSI-/LOH- sporadic CRCs suggest that the majority of these tumors evolve through CIMP. These findings suggest that CIN and CIMP represent 2 independent and inversely related mechanisms of genetic and epigenetic instability in sporadic CRCs and confirm that MSI cancers arise as a consequence of CIMP. PMID- 17087943 TI - Intestinal neurofibromatosis is a subtype of familial GIST and results from a dominant activating mutation in PDGFRA. AB - BACKGROUND & AIMS: Intestinal neurofibromatosis (Online Mendelian inheritance in Man database number 162220) is an alternate form of neurofibromatosis. Patients present with neurofibromas limited to the intestine in the absence of any other typical features of NF1 and NF2. At present, the molecular basis of intestinal neurofibromatosis remains elusive. The aim of the present study was to find the gene responsible for intestinal neurofibromatosis and to characterize functionally the mutation. METHODS: Three candidate genes (NF1, KIT, and PDGFRA) were screened for mutations in 3 sisters diagnosed with intestinal neurofibromatosis. Five tumors were available for pathologic examination. Activation (phosphorylation) of PDGFRalpha was subsequently tested by Western blot analysis on a transfected 293T and Ba/F3 cell line. RESULTS: We found an inherited mutation (Y555C) in the juxtamembrane domain of PDGFRA in the affected individuals. The Y555C mutation leads to autophosphorylation and thus activation of PDGFRalpha. These observations confirm that PDGFRalpha(Y555C) is an oncogenic kinase. The clinical phenotype in the reported family resembles the syndrome of familial gastrointestinal stromal tumors (familial GIST). Somatic activating mutations in KIT and PDGFRA are frequent in sporadic GISTs, and mutations in both genes have also been described in familial GISTs. The tumors in the reported family are morphologically identical to intestinal neurofibromas, but, immunohistochemically, they do not express S100 or any of the known GIST markers. CONCLUSIONS: The inherited PDGFRA mutation in the reported family shows that intestinal neurofibromatosis is allelic to familial GIST caused by PDGRA mutations. We therefore propose that these tumors be classified as familial KIT negative gastrointestinal stromal tumors. PMID- 17087944 TI - Therapeutic treatment of experimental colitis with regulatory dendritic cells generated with vasoactive intestinal peptide. AB - BACKGROUND & AIMS: Crohn's disease is a chronic debilitating disease characterized by severe T helper cell (Th)1-driven inflammation of the colon partially caused by a loss of immune tolerance against mucosal antigens. The use of regulatory dendritic cells (DCs) with the capacity to induce regulatory T cells has been proposed recently for the treatment of Crohn's disease in a strategy to restore immune tolerance. Vasoactive intestinal peptide is an immunomodulatory neuropeptide that induces regulatory DCs. The aim of this study was to investigate the therapeutic effect of vasoactive intestinal peptide induced regulatory DCs (DC(VIP)) in a murine model of colitis. METHODS: We examined the therapeutic action of DC(VIP) in the colitis induced by intracolonic administration of trinitrobenzene sulfonic acid, evaluating diverse clinical signs of the disease including weight loss, diarrhea, colitis, and histopathology. We also investigated the mechanisms involved in the potential therapeutic effect of DC(VIP), such as inflammatory cytokines and chemokines, Th1 type response, and the generation of regulatory T cells. RESULTS: DC(VIP) injection significantly ameliorated the clinical and histopathologic severity of colitis, abrogating body weight loss, diarrhea, and inflammation, and increasing survival. The therapeutic effect was associated with down-regulation of both inflammatory and Th1-driven autoimmune response, by regulating a wide spectrum of inflammatory mediators directly through activated macrophages, and by generating interleukin-10-secreting regulatory T cells with suppressive capacity on autoreactive T cells. CONCLUSIONS: The possibility to generate/expand ex vivo regulatory DC(VIP) opens new therapeutic perspectives for the treatment of Crohn's disease in human beings, and may minimize the dependence on nonspecific immunosuppressive drugs used currently for autoimmune disorders. PMID- 17087945 TI - Saccharomyces boulardii inhibits inflammatory bowel disease by trapping T cells in mesenteric lymph nodes. AB - BACKGROUND & AIMS: Saccharomyces boulardii is a nonpathogenic yeast used for treatment of diarrhea. We used a mice model of inflammatory bowel disease (IBD) to analyze the effects of S boulardii on inflammation. METHODS: Lymphocyte transferred SCID mice, displaying IBD, were fed daily with S boulardii. Weight loss and inflammatory status of the colon were monitored. Nuclear factor-kappaB activity was assessed in the colon. The CD4(+) T-cell production of interferon (IFN) gamma was evaluated by enzyme-linked immunosorbent assay, and a comprehensive reverse-transcription polymerase chain reaction (RT-PCR) analysis for both colon and mesenteric lymph nodes was performed. Finally, we analyzed cell migration mechanisms in vitro and in vivo. RESULTS: S boulardii treatment inhibits IBD. S boulardii induces an accumulation of IFN-gamma-producing T-helper 1 cells within the mesenteric lymph nodes correlated with a diminution of CD4(+) T-cell number and IFN-gamma production by CD4+ T cells within the colon. The influence of S boulardii treatment on cell accumulation in mesenteric lymph nodes was also observed in normal BALB/c mice and involves modifications of lymph node endothelial cell adhesiveness by a yeast secretion product. CONCLUSIONS: S boulardii has a unique action on inflammation by a specific alteration of the migratory behavior of T cells, which accumulate in mesenteric lymph nodes. Therefore, S boulardii treatment limits the infiltration of T-helper 1 cells in the inflammed colon and the amplification of inflammation induced by proinflammatory cytokines production. These results suggest that S boulardii administration may have a beneficial effect in the treatment of IBD. PMID- 17087946 TI - Interferons alpha and lambda inhibit hepatitis C virus replication with distinct signal transduction and gene regulation kinetics. AB - BACKGROUND & AIMS: Hepatitis C virus (HCV) is a major cause of chronic liver disease, cirrhosis, and hepatocellular carcinoma. Current therapy with pegylated interferon alpha (IFN-alpha) in combination with ribavirin is associated with adverse effects and often fails to induce a sustained response. IFN-lambdas, recently discovered IFN gene family members, exhibit antiviral and cell stimulatory activities similar to IFN-alpha. We aimed to determine whether IFN lambda exhibits antiviral activity toward HCV and to compare the signal transduction and effector gene pathways with those of IFN-alpha. METHODS: Using the HCV replicon system and cell culture infectious reporter virus, we compared IFN-alpha and IFN-lambda effects on HCV RNA replication and protein expression, as measured by quantitative reverse-transcriptase polymerase chain reaction, luciferase expression, and Western blot. Receptor expression and signaling pathways were explored using flow cytometry and Western blot. IFN-alpha- and IFN lambda-mediated gene expression changes were compared using microarray analyses. RESULTS: IFN-lambda exhibited dose- and time-dependent HCV inhibition, independent of types I and II IFN receptors. The kinetics of IFN-lambda-mediated signal transducers and activators of transcription (STAT) activation and induction of potential effector genes were distinct from those of IFN-alpha. IFN lambda induced steady increases in levels of known interferon stimulated genes (ISGs), whereas IFN-alpha ISGs peaked early and declined rapidly. IFN-lambda inhibited replication of HCV genotypes 1 and 2 and enhanced the antiviral efficacy of subsaturating levels of IFN-alpha. CONCLUSIONS: These results demonstrate distinct differences in IFN-lambda- and IFN-alpha-induced antiviral states. Understanding these differences may prove useful for developing new HCV treatment strategies. PMID- 17087947 TI - A randomized trial of rofecoxib for the chemoprevention of colorectal adenomas. AB - BACKGROUND & AIMS: In human and animal studies, nonsteroidal anti-inflammatory drugs have been associated with a reduced risk of colorectal neoplasia. Although the underlying mechanisms are unknown, inhibition of cyclooxygenase (COX), particularly COX-2, is thought to play a role. We conducted a randomized, placebo controlled, double-blind trial to assess whether use of the selective COX-2 inhibitor rofecoxib would reduce the risk of colorectal adenomas. METHODS: We randomized 2587 subjects with a recent history of histologically confirmed adenomas to receive daily placebo or 25 mg rofecoxib. Randomization was stratified by baseline use of cardioprotective aspirin. Colonoscopic follow-up evaluation was planned for 1 and 3 years after randomization. The primary end point was all adenomas diagnosed during 3 years' treatment. In a modified intent to-treat analysis, we computed the relative risk of any adenoma after randomization, using Mantel-Haenszel statistics stratified by low-dose aspirin use at baseline. RESULTS: Adenoma recurrence was less frequent for rofecoxib subjects than for those randomized to placebo (41% vs 55%; P < .0001; relative risk [RR], 0.76; 95% confidence interval [CI], 0.69-0.83). Rofecoxib also conferred a reduction in risk of advanced adenomas (P < .01). The chemopreventive effect was more pronounced in the first year (RR, 0.65; 95% CI, 0.57-0.73) than in the subsequent 2 years (RR, 0.81; 95% CI, 0.71-0.93). As reported previously, rofecoxib was associated with increased risks of significant upper gastrointestinal events and serious thrombotic cardiovascular events. CONCLUSIONS: In this randomized trial, rofecoxib significantly reduced the risk of colorectal adenomas, but also had serious toxicity. PMID- 17087948 TI - Cholesterol gallstone susceptibility loci: a mouse map, candidate gene evaluation, and guide to human LITH genes. PMID- 17087949 TI - Hepatitis C and risk of lymphoma: results of the European multicenter case control study EPILYMPH. AB - BACKGROUND & AIMS: Increasing evidence points toward a role of hepatitis C virus (HCV) infection in the etiology of malignant lymphomas. However, previous epidemiologic studies were limited in size to establish an association between HCV infection and specific lymphoma subtypes. We performed a large, multicenter, case-control study to address this question. METHODS: The study comprised 5 European countries and included newly diagnosed cases of any lymphoid malignancy recruited between 1998 and 2004. Controls were matched to cases by 5-year age group, sex, and study center. In-person interviews were conducted to collect data on demographic, medical, and family history as well as environmental exposures. Serum samples of 1807 cases and 1788 controls (excluding human immunodeficiency virus-positive and organ-transplantation subjects) were screened for HCV infection using an enzyme immunoassay. Positive as well as randomly selected negative samples were subjected to HCV RNA detection and HCV genotyping. RESULTS: HCV infection was detected in 53 (2.9%) lymphoma cases and in 41 (2.3%) control subjects (odds ratio [OR], 1.42; 95% confidence interval [CI]: 0.93-2.15). Restricted to individuals who tested positive for HCV-RNA (indicating persistent infection and active viral replication), the OR was 1.82 (95% CI: 1.13-2.91). In subtype-specific analyses, HCV prevalence was associated with diffuse large B cell lymphoma (OR, 2.19; 95% CI: 1.23-3.91) but not with chronic lymphocytic leukemia or follicular, Hodgkin's, or T-cell lymphoma. The sample size was not sufficient to derive any conclusions for rare lymphoma entities such as splenic marginal zone lymphoma. CONCLUSIONS: These results support a model that chronic HCV replication contributes to lymphomagenesis and establish a specific role of HCV infection in the pathogenesis of diffuse large B-cell lymphoma. PMID- 17087950 TI - CD8+CD28- regulatory T lymphocytes prevent experimental inflammatory bowel disease in mice. AB - BACKGROUND & AIMS: Immune responses to innocuous intestinal antigens appear tightly controlled by regulatory T lymphocytes. While CD4+ T lymphocytes have recently attracted the most attention, CD8+ regulatory T-cell populations are also believed to play an important role in control of mucosal immunity. However, CD8+ regulatory T-cell function has mainly been studied in vitro and no direct in vivo evidence exists that they can control mucosal immune responses. We investigated the capacity of CD8+CD28- T cells to prevent experimental inflammatory bowel disease (IBD) in mice. METHODS: CD8+CD28- regulatory T cells were isolated from unmanipulated mice and tested for their capacity to inhibit T cell activation in allogeneic mixed lymphocyte cultures in vitro and to prevent IBD induced by injection of CD4+CD45RB(high) cells into syngeneic immunodeficient RAG-2 mutant mice. RESULTS: CD8+CD28- T lymphocytes inhibited proliferation and interferon gamma production by CD4+ responder T cells in vitro. CD8+CD28- regulatory T cells freshly isolated from spleen or gut efficiently prevented IBD induced by transfer of colitogenic T cells into immunodeficient hosts. Regulatory CD8+CD28- T cells incapable of producing interleukin-10 did not prevent colitis. Moreover, IBD induced with colitogenic T cells incapable of responding to transforming growth factor beta could not be prevented with CD8+CD28- regulatory T cells. CD8+CD28+ T cells did not inhibit in vitro or in vivo immune responses. CONCLUSIONS: Our findings show that naturally occurring CD8+CD28- regulatory T lymphocytes can prevent experimental IBD in mice and suggest that these cells may play an important role in control of mucosal immunity. PMID- 17087951 TI - Long-term therapy with adefovir dipivoxil for HBeAg-negative chronic hepatitis B for up to 5 years. AB - BACKGROUND & AIMS: Treatment with adefovir dipivoxil for 48 weeks resulted in clinical improvement in patients with hepatitis B e antigen (HBeAg)-negative chronic hepatitis B that was lost when treatment was discontinued. We investigated the efficacy, safety, and resistance profile of adefovir dipivoxil treatment for up to 240 weeks. METHODS: HBeAg-negative patients were treated double blind with placebo or adefovir dipivoxil 10 mg once daily for 48 weeks, followed by adefovir dipivoxil from week 49 to 96. At week 97, 125 patients enrolled in a 144-week, open-label phase. Patients received adefovir dipivoxil for up to 192 or 240 weeks. RESULTS: Serum hepatitis B virus (HBV) DNA levels were less than 1000 copies per milliliter in 67% of patients, and alanine aminotransferase (ALT) levels normalized in 69% after 240 weeks. After 192 or 240 weeks of treatment, over 83% of patients had improvement in necroinflammation, and over 73% had improvement in fibrosis. Ishak fibrosis scores improved compared with baseline in 35%, 55%, and 71% of patients after 48, 192, and 240 weeks of adefovir dipivoxil, respectively. After 240 weeks, the cumulative probability of HBV polymerase mutations was 29%, but the cumulative probability of mutations with virologic resistance was 20% and of mutations, virologic resistance, and ALT elevations was 11%. Slight elevations in creatinine were confirmed in 4 (3%) patients. CONCLUSIONS: Treatment with adefovir dipivoxil for up to 240 weeks was well tolerated and produced significant, increasing improvement in hepatic fibrosis, durable suppression of HBV replication, normalization of liver enzymes, and delayed development of resistance. PMID- 17087952 TI - Gastric sensorimotor functions and hormone profile in normal weight, overweight, and obese people. AB - BACKGROUND & AIMS: Peptide YY (PYY) levels are reported to be decreased in obesity. The relation between gastric functions, satiation, and gut hormones in obesity is incompletely understood. The aim of this study was to compare gastric volumes, emptying, maximum tolerated volumes, postchallenge symptoms, and selected gut hormones in normal, overweight, or obese healthy volunteers. METHODS: In 73 nonbulimic normal, overweight, or obese participants weighing less than 137 kg, we measured gastric emptying of solids and liquids by scintigraphy (gastric emptying half-time [GE t(1/2)]); gastric volumes by single-photon emission computed tomography; maximum tolerated volumes and symptoms by satiation test; and plasma leptin, ghrelin, insulin, glucagon-like peptide 1, and PYY levels. Groups were compared using 1-way analysis of covariance adjusted for sex. Univariate associations among measured responses were assessed using Spearman correlations. Multiple linear regression models, adjusting for weight and sex, assessed the independent ability of gastric functions and hormones to predict satiation volume. RESULTS: Obese and overweight subjects had significantly lower postprandial gastric volumes, higher fasting and postprandial insulin and leptin levels, and lower fasting ghrelin and lower postprandial reduction in ghrelin levels. PYY levels were not different in obese or overweight subjects compared with controls. The GE t(1/2) was correlated inversely with postprandial PYY; increased body weight was associated with faster GE t(1/2) of solids (r(s) = 0.33, P = .005) and liquids (r(s) = 0.24, P = .04). Postprandial changes in gastric volume and PYY were independent predictors of satiation (both P = .01). CONCLUSIONS: Overweight or obesity are associated with lower postprandial gastric volumes and normal PYY levels. Gastric emptying influences postprandial PYY levels. Postprandial PYY and gastric volume independently predict satiation volume in nonbulimic people across a wide body mass index range. PMID- 17087953 TI - Fracture risk in people with primary biliary cirrhosis: a population-based cohort study. AB - BACKGROUND & AIMS: Controversy exists as to whether people with primary biliary cirrhosis (PBC) have an increased risk of developing osteoporosis and the extent to which this may translate into an increased risk of fracture. We have performed a cohort study using the General Practice Research Database to quantify the excess fracture risk in people with PBC. METHODS: We identified 930 people with PBC and 9202 age- and sex-matched control subjects. We used Cox regression to estimate the hazard ratios for any fracture, hip fracture, and ulna/radius fracture in the PBC cohort compared with the general population. RESULTS: There were approximately 2-fold relative increases in the risk of any fracture, hip fracture, and ulna/radius fracture for the PBC cohort compared with the general population (hazard ratio [HR], 2.03; 95% confidence interval [CI]: 1.70-2.44; HR 2.14 (95% CI: 1.40-3.28), and HR, 1.96; 95% CI: 1.42-2.71, respectively). The absolute excess in fracture rates were for any fracture, 12.5 per 1000 person years (95% CI: 8.1-16.9); for hip fracture, 1.9 per 1000 person-years (95% CI: 0.3-3.5); and for ulna/radius fracture, 3.4 per 1000 person-years (95% CI: 1.2 5.7). In those people with more severe disease, the relative risks of fracture were similar (any fracture HR, 2.24; hip fracture HR, 1.25; ulna/radius fracture HR, 1.28). CONCLUSIONS: There are modest increases in both the absolute and relative fracture risks in people with PBC compared with the general population, with the excess risks similar in those with more severe disease. PMID- 17087954 TI - Correlation between the single-site CpG methylation and expression silencing of the XAF1 gene in human gastric and colon cancers. AB - BACKGROUND & AIMS: X-linked inhibitor of apoptosis protein (XIAP)-associated factor 1 (XAF1) antagonizes the anti-caspase activity of XIAP. XAF1 messenger RNA is present in normal tissues but undetectable in various cancers and thus poses a potential tumor suppressor gene. The aim of this study was to examine the novel pattern of methylation of XAF1 in gastric and colon cancers and locate the important CpG sites for transcriptional regulation and tumor progression. METHODS: XAF1 expression was detected by reverse-transcription polymerase chain reaction (PCR) and Western blot analysis. Four different fragments around the transcription start site of XAF1 were cloned and examined putative promoter activities by luciferase reporter assay. Each CpG site in fragment F291 was mutated by site-directed mutagenesis technique, and the change of promoter activity of this fragment was detected by luciferase reporter assay. Methylation status of XAF1 was determined by methylation-specific PCR (MSP) and bisulfite DNA sequencing PCR analysis. RESULTS: Down-regulation of XAF1 in association with hypermethylation was detected in 3 of 4 human gastric cancer cell lines and 6 of 8 colon cancer cell lines. Of the 4 promoter fragments, F291 showed the highest promoter activity, which could be down-regulated obviously by the mutation of particular CpG sites. Moreover, aberrant hypermethylation of these important CpG sites was strongly associated with the development of gastric and colon cancers. CONCLUSIONS: A cluster of methylated CpG sites instead of CpG islands located in the promoter area resulted in gene silencing of XAF1, and CpGs at -2nd, -1st, and +3rd positions are functionally more important in its transcriptional regulation. PMID- 17087955 TI - Contribution of sensory neurons to sex difference in the development of stress induced gastric mucosal injury in mice. AB - BACKGROUND & AIMS: Sensory neurons play a critical role in reducing stress induced gastric mucosal injury by releasing calcitonin gene-related peptide (CGRP) through an increase in gastric mucosal levels of prostacyclin (PGI(2)). Because estrogen enhances nerve growth factor-mediated CGRP production in sensory neurons, we hypothesized that stress-induced gastric mucosal injury occurs less in females than in males. METHODS: Gastric ulcer index, gastric myeloperoxidase activity, and gastric tissue levels of CGRP and 6-keto-PGF(1alpha), a stable metabolite of PGI(2), were determined in male and female wild-type (CGRP(+/+)) mice and CGRP knockout (CGRP(-/-)) mice subjected to water-immersion restraint stress. RESULTS: In CGRP(+/+) mice, ulcer index and myeloperoxidase activities were lower and gastric tissue levels of CGRP and 6-keto-PGF(1alpha) were higher in female mice than in male mice, but there were no such sex differences in CGRP( /-)mice. Sex differences in CGRP(+/+) mice were eliminated by pretreatment with SB366791 (500 microg/kg intraperitoneally), a vanilloid receptor antagonist, and by ovariectomy. Reversal of sex differences by ovariectomy was not observed in female CGRP(+/+) mice with estradiol replacement (1 mg . kg(-1). wk(-1) for 3 weeks). Levels of CGRP messenger RNA in dorsal root ganglion neurons isolated from female CGRP(+/+) mice were decreased by ovariectomy, and these decreases were reversed by estradiol replacement. CONCLUSIONS: Estrogen-mediated increases in CGRP levels in sensory neurons might contribute to reduce stress-induced gastric mucosal injury by attenuating inflammatory responses. This might at least partly explain the sex difference observed in the development of stress-induced gastric mucosal injury in mice. PMID- 17087956 TI - Dietary factors and biomarkers involved in the methylenetetrahydrofolate reductase genotype-colorectal adenoma pathway. AB - BACKGROUND & AIMS: Methylenetetrahydrofolate reductase (MTHFR) is involved in intracellular folate homeostasis and metabolism. We assessed 2 polymorphisms in the MTHFR gene (C677T and A1298C) in relation to colorectal adenoma recurrence and conducted analyses to investigate their joint effects with plasma and dietary markers of folate status. METHODS: We prospectively analyzed data from 1598 individuals genotyped for the C677T polymorphism and 1583 with data on A1298C. RESULTS: Among nonusers of multivitamin supplements, compared with wild-type carriage, higher odds of recurrence were observed for those with the 677 TT variant (odds ratio [OR], 1.66; 95% confidence interval [CI], 1.04-2.63) and a nonsignificant increase was observed among those with the 1298 CC variant (OR, 1.50; 95% CI, 0.93-2.40). Diplotype analyses among nonusers of multivitamins showed that individuals who carry the MTHFR 677TT_1298AA or 677CC_1298CC combination were significantly more likely to have a recurrence compared with those with the double wild-type (OR, 2.05 for TT_AA and 1.85 for CC_CC). Higher odds of recurrence were observed among participants with low folate intake or plasma folate and the 677 TT or 1298 CC variants compared with those with lower levels and the wild-type or heterozygous genotypes. Stronger associations were shown for the combination of high homocysteine and the 677 TT variant (OR, 2.29; 95% CI, 1.00-5.26) but not the 1298 CC variant (OR, 1.09; 95% CI, 0.39-3.01). CONCLUSIONS: We propose that the effect of the MTHFR genotypes on increasing risk of adenoma recurrence in the presence of a low folate status is through their increase in homocysteine concentrations, which in turn could result in DNA hypomethylation via pathways involving S-adenosylhomocysteine. PMID- 17087957 TI - Transient lower esophageal sphincter relaxations and reflux: mechanistic analysis using concurrent fluoroscopy and high-resolution manometry. AB - BACKGROUND & AIMS: The aim of this study was to perform a detailed analysis of the mechanics leading to esophagogastric junction (EGJ) opening during transient lower esophageal sphincter relaxations (tLESRs) using high-resolution manometry coupled with simultaneous fluoroscopy. METHODS: Six subjects without hiatus hernia had endoclips placed at the squamocolumnar junction and 10 cm proximal. A 36-channel solid-state manometric assembly was placed spanning from stomach to pharynx, and subjects were studied for 2 hours after a high-fat meal. An esophageal pH electrode also was placed and fluoroscopy was initiated at the onset of a tLESR. Axial clip movement was measured during replay of the videotaped fluoroscopy and was correlated with manometric data. RESULTS: Ninety three tLESRs were recorded, 62 tLESRs of which had good fluoroscopic visualization. Seventy-eight tLESRs had manometric evidence of flow and the majority had evidence of a common cavity (88%), but few were detected by the pH electrode. Esophageal shortening and crural diaphragm inhibition always preceded EGJ opening and common cavity. A positive pressure gradient between the stomach and the EGJ lumen of 7.1 mm Hg (interquartile range, 4.1-9.1 mm Hg) preceded the EGJ opening. CONCLUSIONS: Key events leading to the EGJ opening during tLESRs were LES relaxation, crural diaphragm inhibition, esophageal shortening, and a positive pressure gradient between the stomach and the EGJ lumen. The manometric signature of opening was pressure equalization within the EGJ, but this only occasionally was associated with pH evidence of reflux. Future investigations will need to analyze how this delicately balanced anatomic-physiologic system is perturbed in subjects with reflux disease. PMID- 17087958 TI - The use of factor VIIa in haemorrhagic shock and intracerebral bleeding. AB - Factor VIIa is a revolutionary new pharmaceutical that promises to change the anaesthesia and critical care approach to major trauma. It is an extremely potent pro-coagulant agent, and while it enables haemostasis at the site of tissue injury, it also has the possibility of producing life-threatening thromboembolic complications. New data regarding FVIIa use is published almost every month, leading to a rapidly evolving clinical understanding of the potential indications, and potential pitfalls, of off-label use. Determination of appropriate practice, including the ability to judge the risks and benefits of FVIIa therapy for individual cases, is still some years in the future, and will depend in large part on clinical trials which are just getting underway. PMID- 17087959 TI - Outcome prediction in trauma. AB - BACKGROUND: In the Trauma Audit and Research Network (TARN), currently the largest trauma network in Europe, outcome prediction is performed using the TRISS methodology since 1989. Its database contains 200,000 hospital admissions from 110 hospitals over the country, but a large amount of data is lost for the modelling because of missing data. To improve some of the shortcomings of TRISS a new model was developed. METHODS: The data for modelling consisted of 100,399 hospital trauma admissions over the period 1996 to 2001. Using the Glasgow Coma Score (GCS) instead of RTS has dramatically reduced the number of missing cases. Gender and its interaction with age have also been included in the model. The model was tested on different subsets of cases traditionally excluded, such as children, those with penetrating injuries, and ventilated and transferred patients. The new model included all those subsets using age, a transformation of ISS, GCS, gender and gender by age interaction as predictors. RESULTS: The model has shown a good discriminant ability tested by the area under the receiver operating characteristic (AROC) curve. The values of the AROC for the new model were 0.947 (95% CI: 0.943-0.951) on the prediction set and 0.952 (95% CI: 0.946 0.957) on the validation set compared respectively with 0.937 (95% CI: 0.932 0.943) and 0.941 (95% CI: 0.936-0.952) for TRISS. CONCLUSION: The new model has enabled us to include most of the cases that were excluded under the TRISS's inclusion criteria, less missing data are incurred and the predictive performance was significantly better than that of the TRISS model as shown by the AROC curves. PMID- 17087960 TI - Evaluation and outcome of patients after polytrauma--can patients be recruited for long-term follow-up? AB - INTRODUCTION: There is limited information available about the long-term follow up of polytrauma patients. In this study, the social and medical sequelae of trauma were reinvestigated at 10 years after the injury. METHODS: Patients were selected out of a population of polytraumatised patients treated at Hannover Medical School between 1973 and 1990. INCLUSION CRITERIA: multiple injuries treated at one institution, age between 3 and 60 years of age at the time of injury. Patients were recruited by gathering their residences from the charts. If patients had moved, up to three different registration offices were contacted by mail. The patient was contacted by mail (maximum three times) and by telephone. A patient was documented as lost to follow-up if none of these attempts was successful, or if he did not fulfil three subsequent appointments. All patients were examined by a physician, using a patient questionnaire and a standardized physical exam. RESULTS: Six hundred thirty-seven patients (67.8% of the potential enrollees) were evaluated on an outpatient basis by a trauma surgeon using a self administered patient questionnaire and a standardized physical exam. In these, the average follow-up was 17.5 (range 10-28) years; the average Injury Severity Score (ISS) was 20.7 (range 4-54). Head injuries were the third most frequent injuries, but represented the most frequent cause of permanent disability (40%). The overall rehabilitation status graded by the patients was very good in 14.1%, good in 33.0%, satisfactory in 29.3%, sufficient in 16.0% and poor or insufficient in 7.5%. CONCLUSIONS: This study suggests that a high percentage of patients can be recruited for follow-up even after 10 years post trauma with the use of a meticulous reinvitation strategy. Head injuries accounted for the most frequent cause of disability, suggesting that more research should be provided to minimise the degree of injury and improve the outcome for head injured patients. Subjective grading of the outcome was better than expected in patients who had regained complete social rehabilitation. PMID- 17087961 TI - Assessing outcomes in paediatric trauma populations. AB - INTRODUCTION: Assessing outcomes in the paediatric trauma population is important. Identifying suitable instruments can be problematic. This article highlights the commonly used outcome measures for assessing functional status and health related quality of life in paediatric trauma patients. Child specific characteristics which impact upon instrument development and selection are reviewed. METHODS: An electronic database search was conducted to identify suitable English language measures used for outcome assessment in paediatric trauma patients from 1966 to present. RESULTS: Nine suitable instruments were identified, the child health questionnaire (CHQ), Glasgow outcome scale (GOS), paediatric overall performance category (POPC), PedsQL 4.0 generic core scales, paediatric evaluation of disability inventory (PEDI), functional independence measure (FIM), WeeFIM and an unnamed paediatric trauma specific measure [Gofin R, Hass T, Adler B, The development of disability scales for childhood and adolescent injuries. J Clin Epidemiol 1995;48:977-84]. Each instrument was found to have advantages and disadvantages for assessing outcomes in a paediatric trauma population. CONCLUSION: The PedsQL 4.0 generic core scale could be feasible for administration as a routine outcome measure for paediatric trauma groups. For very young children an additional measure such as that proposed by Gofin et al. [Gofin R, Hass T, Adler B, The development of disability scales for childhood and adolescent injuries. J Clin Epidemiol 1995;48:977-84] may be indicated. Future use of these instruments in the paediatric population would benefit from further psychometric evaluation. PMID- 17087962 TI - The importance of quality of survival as an outcome measure for an integrated trauma system. AB - Risk-adjusted survival rates have been the principle mode of comparison between trauma systems. In mature trauma systems, it is possible that there will be further improvements in survival but these are likely to be small. In the future, the largest gains will come from quality of life and improved function of the survivors. The issues related to measuring quality of survival for trauma systems are reviewed, including feasibility, ethical considerations, risk adjustment of outcomes of survivors, and challenges for selection of instruments and administration. In addition, the preliminary experiences of measuring outcomes in survivors through the Victorian State Trauma Registry are discussed. Although function and quality of life have been identified as important factors to measure in trauma populations, a standardised protocol has not been established. The experience in Victoria suggests that monitoring of population-based outcomes in survivors is feasible and may create the basis for benchmarking the level of morbidity in survivors. PMID- 17087963 TI - Clathrin-dependent pathways and the cytoskeleton network are involved in ceramide endocytosis by a parasitic protozoan, Giardia lamblia. AB - Although identified as an early-diverged protozoan, Giardia lamblia shares many similarities with higher eukaryotic cells, including an internal membrane system and cytoskeleton, as well as secretory pathways. However, unlike many other eukaryotes, Giardia does not synthesize lipids de novo, but rather depends on exogenous sources for both energy production and organelle or membrane biogenesis. It is not known how lipid molecules are taken up by this parasite and if endocytic pathways are involved in this process. In this investigation, we tested the hypothesis that highly regulated and selective lipid transport machinery is present in Giardia and necessary for the efficient internalization and intracellular targeting of ceramide molecules, the major sphingolipid precursor. Using metabolic and pathway inhibitors, we demonstrate that ceramide is internalized through endocytic pathways and is primarily targeted into perinuclear/endoplasmic reticulum membranes. Further investigations suggested that Giardia uses both clathrin-dependent pathways and the actin cytoskeleton for ceramide uptake, as well as microtubule filaments for intracellular localization and targeting. We speculate that this parasitic protozoan has evolved cytoskeletal and clathrin-dependent endocytic mechanisms for importing ceramide molecules from the cell exterior for the synthesis of membranes and vesicles during growth and differentiation. PMID- 17087964 TI - O-methylated glycans from Toxocara are specific targets for antibody binding in human and animal infections. AB - The parasitic helminth Toxocara canis is a widely distributed nematode of mammals. Larval parasites, which infect a wide range of hosts including mice and humans, export glycosylated macromolecules bearing novel methylated oligosaccharide structures, similar to the mammalian blood group antigen H but bearing one or two O-methylated substitutions on the terminal fucose and subterminal galactose residues. We have studied the reactivity of synthetic forms of these glycans to parasite-specific antibodies and mammalian immune system lectins. Murine antibodies, generated to T. canis infection, predominantly recognise the mono-O-methylated form with the beta-configuration of the GalNAc residue (MoMbeta), and antibodies are entirely IgM isotype. The mAb Tcn-2 reproduces this pattern, and shows little reactivity to either the alpha isomer (MoMalpha) or the di-O-methylated form (DiM). Antibodies generated to helminth infections other than T. canis were unreactive with the glycans, except antibodies to other members of the Toxocara genus. Hence, the carbohydrate structures represent immunogenic, genus-specific antigens. Antibodies from human toxocariasis patients are reactive with the same sugars, although preferentially towards DiM. Sera from unrelated helminth infections do not react, confirming the status of these structures as Toxocara-specific glycans. The human dendritic cell lectin, DC-SIGN, was found to bind both Toxocara excretory/secretory products and mammalian blood group antigen H3. However, DC-SIGN did not bind the synthetic glycans, indicating additional non-methylated carbohydrates may also play a role in the interaction between T. canis and its host. PMID- 17087965 TI - Adeno-associated virus LPL(S447X) gene therapy in LDL receptor knockout mice. AB - BACKGROUND: Overexpression of lipoprotein lipase (LPL) protects against atherosclerosis in genetically engineered mice. We tested whether a gene therapy vector that delivers human (h) LPL(S447X) cDNA to skeletal muscle could induce similar effects. METHODS: LDL receptor knockout (LDLr-/-) mice were injected intramuscular (i.m.) with adeno-associated virus serotype 1 (AAV1) LPL(S447X) or PBS. Four weeks later they were started on an atherogenic diet for 12 weeks. After termination, atherosclerosis was assessed and homogenates of muscle and liver tissue were analyzed. RESULTS: AAV1-treated mice showed hLPL concentrations of 768+/-293 ng/mL in post-heparin plasma associated with 48% reductions of fasting triglycerides (TG) levels (p<0.0001). In the absence of an effect on total cholesterol (TC) levels, no effects on atherosclerosis were found. An increase in lipid content of injected muscles was accompanied by a significant decrease of TG (-20%, p<0.0001) and free cholesterol (FC) content (-24%, p<0.0001) in liver homogenates. CONCLUSIONS: The data show that transgenic hLPL(S447X) on top of endogenous murine LPL reduces fasting TG levels in plasma but has no effect on atherosclerosis in LDLr-/- mice. While lipid accumulation in the injected muscle was anticipated, this coincided with an interesting decrease of both TG and FC in liver homogenates. PMID- 17087966 TI - The effect of interleukin-10 knock-out and overexpression on neointima formation in hypercholesterolemic APOE*3-Leiden mice. AB - OBJECTIVE: Inflammatory factors are thought to play a regulatory role in restenosis. Interleukin-10 (IL10) is an important anti-inflammatory cytokine with anti-atherogenic potentials. The aim of this study was to assess the effects of IL10 modulation on cuff-induced neointima formation in hypercholesterolemic APOE*3-Leiden mice. METHODS: The involvement of IL10 in neointima formation was studied in a hypercholesterolemic mouse model of cuff-induced stenosis of the femoral artery by IL10 knocking-out or overexpression procedures. IL10(+/-) mice were crossbred with APOE*3-Leiden mice to generate hypercholesterolemic APOE*3 LeidenIL10(-/-) mice. To achieve IL10 overexpression in APOE*3-Leiden mice, a single intramuscular injection of a murine IL10 overexpression plasmid was performed followed by electroporation. RESULTS: Knocking-out IL10, in hypercholesterolemic APOE*3-Leiden mice, resulted in a significant 1.9-fold increase of neointima surface as compared to APOE*3-LeidenIL10(+/+) littermates (p=0.02). Conversely, a marked 45% inhibition on cuff-induced neointima formation was obtained after IL10 overexpression (p=0.02). Electrodelivery of IL10 vector leads to detectable IL10 serum levels, with a sustained expression over the experimental period of 3 weeks. IL10 overexpression reduced plasma cholesterol levels in APOE*3-Leiden mice, whereas IL10 deficiency in these mice did not lead to altered cholesterol levels as compared to the IL10(+/+) group. Finally, IL10 overexpression stimulated endogenous IL10 mRNA expression in the spleen and reduced the transcriptional responses of several pro-inflammatory cytokines. CONCLUSION: Here, we clearly demonstrate the role of IL10 in the development of neointima formation in hypercholesterolemic mice and the potential therapeutic effect of non-viral electrodelivery of IL10 cDNA to inhibit post-angioplasty restenosis. PMID- 17087967 TI - Serum gamma-glutamyl transferase activity: a new marker for stent restenosis? AB - Stent restenosis remains the main limitation of percutaneous coronary intervention. Elevated serum gamma-glutamyl transferase (GGT) level is associated with an inflammatory response. We aimed to determine the correlation of stent restenosis with the serums level of GGT. One hundred and twenty patients (age 58.56+/-10.46 years, 66% male) with a history of coronary stent implantation and had undergone control coronary angiography (60 with restenosis and 60 without) were included. All had baseline serum GGT activity and were free of systemic and hepatobiliary disease. Median baseline serum GGT activity was significantly higher in patients with restenosis (34.00 U/L (24.00-47.75)) than in those without restenosis (21.00 U/L (17.25-26.7500)) (P<0.0001). Stent restenosis was identified in 38% of the patients with a serum GGT value >40 U/L and in 5% of patients with a serum GGT value